PMID- 16370606 TI - The relationship of CPI measures of interpersonal adequacy to safety and esteem motives. AB - This study was concerned with establishing the relationship of CPI measures of interpersonal adequacy to SCT measures of safety and esteem motives. A large number of students were given the SCT and 56 safety-oriented and 56 esteem oriented subjects were selected and and given the dominance (Do), capacity for status (Cs), self-acceptance (Sa) and sense of well-being (Wb) subscales of the CPI. As predicted the results indicated that esteem-oriented subjects scored higher on all the CPI measures of interpersonal adequacy. PMID- 16370607 TI - Factor and discriminant analysis of the Bem Sex-Role Inventory. AB - Several studies have recently recommended that items which fail to load substantially (< .40) on one of several primary factors characterizing the Bem Sex-Rote Inventory (BSRI) could be deleted without substantial loss of interpretability. This recommendation is questioned on the basis that the extent of interrelatedness of the items is an insufficient indicator of the contribution each item makes towards the classification (identifying sex-types by Bem's median split technique) efficiency of the inventory. Empirical support of this contention was provided by conducting several discriminant analyses (predicting sex-types) on a sample of 1117 BSRI protocols and excluding the various items suggested by the earlier research. These results were then compared with the results of a discriminant analysis using the 40 masculine and feminine items. The analyses using less. than all 40 items produced substantially poorer discrimination than when all 40 items were included. The results Suggest that decisions to exclude items on the basis of factor analysis may be inappropriate and risk serious loss of power in classifying individuals by sex-type. PMID- 16370608 TI - On quantifying the magnitude of sex-role endorsement. AB - The conception of sex-role attributes as multidimensional has led to the development of the median split and various difference score approaches for assigning individuals to sex-rote categories. In response to the criticisms of information loss and imprecision when applying such methods, this report presents a data manipulation (medz) which combines the masculine and feminine scales in an additive rather than subtractive manner, upholds the continuous nature of the variables, and utilizes the median split for categorization. Comparisons of this technique with the t ratio, difference/median split, and direct difference approaches indicate greater differentiation among androgynous subjects when assessed according to the medz. An application of the process and some rationale for its implementation are provided. PMID- 16370616 TI - Earliest childhood memories: four theoretical perspectives. AB - Four separate lines of thought relevant to a theory of early memories (EMs) have appeared in the literature--Freud's, Adler's, an ego psychology view and one adapted from memory theory, Each view was presented in turn and discussed in terms of its implications for assessment and treatment. It was pointed out that none of these four theories addresses several issues relevant to the individual who plans to do research with EMs. PMID- 16370617 TI - Fine points aside: data are data. AB - Movement responses by 5 1/2- to 6 1/2-year-old children on the Holtzman Inkblot Test were reported on by Lockwood et al. in volume 45 of this Journal. Criticism of the findings discussed in that article by J. Swartz, Reinehr, and C. Swartz was published in the December, 1981, volume of the Journal. The present article rebuts their criticism and emphasizes the significance of the original findings which reflect new and highly discrepant normative M response data from that previously available. PMID- 16370618 TI - Rorschach from quality: a comparison of the Exner and Beck systems. AB - The Beck and Exner systems of assigning Rorschach form quality were compared, There are some inherent differences in the form quality tables of these two Rorschach systematizers. The Beck system is more extensive and seems slightly biased toward-or "poor" form quality in comparison to the Exner system. When the Rorschach records of psychiatric patients were scored for form quality, the Beck system yielded a significantly lower Extended F+% than the Exner system. This effect is probably related to the inherent differences in the respective form quality tables which resulted from the different procedures used in deriving these listings of form quality. PMID- 16370619 TI - Anality and routine. PMID- 16370620 TI - MMPI scale development methodology reconsidered. AB - Levitt's (1978) critique of my earlier suggestions regarding MMPI scale development methodology (Clopton, 1978b) correctly noted that scales have been developed by a variety of methods in addition to empirical scale construction, but two of Levitt's other criticisms were mistaken. After Levitt's criticisms are considered, some additional suggestions are provided regarding the empirical development of new MMPI scales. PMID- 16370621 TI - Psychological androgyny and social desirability. AB - The concept of psychological androgyny, because of its implications for sex roles, social change, and human development, is presently of special theoretical interest. Psychologically androgynous persons, as identified by the Bern Sex Role Inventory (BSRI), score higher on both its femininity and masculinity scales. Given the psychometric properties of the BSRI, it can be hypothesized that androgynous persons will have higher social desirability scores. Yet, the "masculine male" and the "feminine female" may be the most socially desirable, in that they conform to normative expectations. Consequently it can also be hypothesized that individuals who are either masculine-typed males or feminine typed females on the BSRI will have the higher social desirability scores. Neither prediction was found to be entirely accurate; instead, androgynous and feminine-typed individuals were found to have higher social desirability scores. Moreover, the data reveal that the BSRI femininity scale has social desirability characteristics which are sex-specific. PMID- 16370622 TI - Ethnicity as a variable in responses to California Psychological Inventory items. AB - The responses of Black and White subjects to the California Psychological Inventory items were compared. On the average, 34% in each of the 18 scales showed significant differences. Approximately two-thirds of the significant items for the males were also significant for the females. Blacks responded in the keyed direction for over 23% of the items. The results of this study were consistent with the results of previous studies. The implications of the results of this research are discussed. It is suggested that future development of tests of this nature carefully consider the socio-cultural factors that might contribute to differential responses to test items on objective personality tests as assessment devices. Scores obtained by Blacks on the CPI should be interpreted against the background of the results of this study. PMID- 16370623 TI - Personality characteristics of ski instructors and predicting teacher effectiveness using the PRF. AB - Normative data for the Personality Research Form (PRF) were collected from 118 ski instructors (40 female, 78 male). Subjects differed significantly from PRF norms on 17 scales (.0001 < or = p < or = .05), thus reiterating the need for caution when applying college-normed summary statistics to non-college populations. The efficacy of PRF scales as predictors of teaching effectiveness was also investigated. Three PRF scales (Abasement, Succorance and Sentience) were negatively correlated with increasing levels of teaching effectiveness. Univariate regressions yielded Abasement and Sentience scores as significant predictors of effective ski instruction. PMID- 16370624 TI - Effects of diagnostician prestige and sex upon subjects' acceptance of genuine personality feedback. AB - Examined subjects' acceptance of genuine, test-based personality feedback as a function of diagnostician prestige and sex, subject sex, and diagnostician. It was found that subjects of both sexes rated the feedback provided by female diagnosticians as significantly more accurate than that provided by male diagnosticians. This was attributable in part to a tendency for subjects to greatly devalue the feedback provided, by low prestige, male diagnosticians, Support for the generalizability of the present results from the social psychological literature was noted and suggestions for future research were provided. PMID- 16370635 TI - Masters of shadows. AB - The interchange of views between Hans Binder and Bruno Klopfer regarding their evaluation and scoring systems for shading responses is traced and their single personal meeting much later is described. PMID- 16370625 TI - Personality types as measured by the 16 PF. AB - The possibility of isolating personality types using 16 PF profiles was studied by utilizing a Q-type factor analytic procedure. Four modal profiles, characteristic of major clusters of subjects, were derived and cross validated. These four major types successfully classified approximately 2/3 of the 16 PF protocols (n = 1037), and classified similar proportions of previously published profiles. Variation of classification rates as a function of race and sex was described, and strategies for utilizing the profiles for research and interpretive purposes were outlined. PMID- 16370636 TI - A cross-cultural comparison of the developmental items of five ethnic groups in the southwest. AB - A population of 3,067 Mexican-American, Pueblo Indian, Navajo Indian, Black, and Anglo elementary children were administered the Draw-A-Person for norming and comparison against the existing Koppitz scoring procedure. A significant number of items for each of the five ethnic groups changed categories with final score interpretation affected. A significant difference in percentage of items drawn by the five ethnic groups was also found, as were regional differences on certain items. PMID- 16370637 TI - The Franck Drawing Completion Test: a tool for research in sex-role identification. AB - The criterion validity of the Franck Drawing Completion Test (FDCT) was reexamined, employing a developmental sample of 486 students from grades 2,4, 6, 8, 10 and 12 as well as an adult sample of 107 college students. The measure was found to differentiate between males and females at a high level of significance at every grade level and in the adult sample. Clear developmental trends were found for males in the general direction of increasing numbers of masculine responses with age, while female means were stable throughout. Results are interpreted as providing strong support for the FDCT as a useful measure of sex role identification. The original theoretical interpretation of the FDCT is reviewed and an alternative values interpretation is suggested. PMID- 16370638 TI - The anal personality: self-disclosure, negativism, self-esteem, and superego severity. AB - Psychoanalytic implications of anal characterology were operationalized, and an experimental situation devised to test hypotheses of various aspects of interpersonal behavior. Subjects selected for the study had been found to score either high or low on Kline's (Ai3) Anality Scale. Self-disclosure and disclosure reciprocity were shown to be negative functions of anality: productivity and superego measures were also shown to be functions of anality. Self-esteem and socio-economic status did not relate to anality levels, while the hypothesis linking anality with negativism was only partially confirmed. Implications for psychoanalytic and social psychology research are discussed. PMID- 16370639 TI - Intellectual ability correlates of the Marlowe-Crowne Social Desirability Scale. AB - The hypothesis that the Marlowe-Crowne Scale would be negatively related to a variety of intellectual ability measures was investigated in a sample of college students. As predicted, MC scores were negatively correlated with measures of intelligence (Quick Test), associational ability (Remote Associates Test), and academic achievement (course grades). These results suggest that the vulnerable self-esteem and defensiveness traditionally ascribed to high versus low MC scorers may at least in part, reflect a history of poorer performance in intellectual and academically related activities. PMID- 16370640 TI - Three measures of locus of control: what do they measure? AB - The Rotter, Adult Nowicki-Strickland, and Levenson IE scales were completed by 175 undergraduate students. Factor analyses reconfirmed the factor structure of the Levenson IE scale. Partial correlation analyses showed that the portion of common variance shared by the Rotter IE and the Adult Nowicki-Strickland IE scales associated closely with Levenson's Chance factor. These findings support a multi-dimensional view of locus of control attribution. PMID- 16370641 TI - A conceptualization and measure of argumentativeness. AB - A conceptualization and measure of argumentativeness is reported. Argumentativeness was conceptualized as a trait Which is composed of the tendency to approach arguments and the tendency to avoid arguments. A 20-item, self-report scale was developed in a series of three factor analytic studies. The internal consistency and test-retest reliabilities of the scale were high. A series of additional studies yielded considerable evidence as to the validity of the scale. The conceptualization of argumentativeness accounts for state as well as trait components. The state components consisted of the individual's perceptions of the probability and importance of success and failure in a particular argumentative situation. A hypothesis was supported which posited response to a particular argument would be predicted better by trait and state components together than by the trait component alone. PMID- 16370654 TI - The Holtzman Inkblot Technique is not the Rorschach: a reply to Lockwood, Roll, and Mathews. AB - A previous article by Lockwood, Roll, and Matthews (1981) reported dramatic differences between the Movement scores obtained by 6-year-old children on the Holtzman Inkblot Technique (HIT) and previously reported normative data for this variable. The Lockwood et al. data were rescored by experienced HIT scorers; their findings-and other possible reasons for the high HIT Movement scores reported by Lockwood et al.-are described and discussed. Finally, the practice of using the HIT in the manner of the Rorschach is criticized. PMID- 16370655 TI - Sex-role-related differences in self-concept and mental health. AB - Examined the differences in serf-concept and psychological health between androgynous, sex-typed, cross-sex-typed, and undifferentiated males and females. Two hundred forty-nine students who were enrolled in introductory psychology classes were classified according to sex type on the basis of their scores on the Bem Sex-Role Inventory. Nine aspects of self-concept and five aspects of psychological health, obtained, from the Tennessee Self-Concept Scale, were analyzed as a function of Sex and Sex Type. A series of analyses of variance and subsequent individual comparisons across groups revealed a clear pattern wherein androgynous subjects manifested more positive self-concepts and more positive psychological health than sex-typed subjects. Undifferentiated subjects manifested the poorest self-concepts, while results for sex-typed and cross-sex typed subjects varied more as a function of the sex of the subject. The data are discussed as appearing to be consistent with Bem's hypothesis that androgynous people may represent a more appropriate societal definition of mental health than strongly sex-typed people. Qualifications of this support are also discussed. PMID- 16370656 TI - Assessment of sex-role stereotyping among male and female psychologist practitioners. AB - Seventy-seven female and 86 male psychologist practitioners filled out the Bem Sex Rote Inventory to describe either a healthy adult male, healthy adult female, or healthy adult, sex unspecified. Analyzing the data according to Bem's classification of masculinity, femininity, and androgyny produced a significant Scale X Condition interaction with male and female practitioners ascribing significantly more masculine than feminine traits to healthy adult men, yet displaying no comparable differences when rating healthy adult women. Analyzing the data according to Spence's classification of masculine, feminine, androgynous, and undifferentiated produced a Group X Condition interaction with practitioners ascribing significantly more masculine traits to healthy adult men and significantly more feminine traits to healthy adult women. Reanalyses omitting "masculine" and "feminine" from Bem's fist of sex-typed traits produced nonsignificant results. The relationship of this to recent criticisms of the construct validity of the BSRI and to the validity of earlier results of therapist sex-role bias was discussed. PMID- 16370657 TI - A data-based analysis of the psychometric performance of the Personality Inventory for Children (PIC): an alternative to the Achenbach review. AB - This paper, in response to Achenbach's review of the Personality Inventory for Children manual and actuarial interpretive guide recently published in the Journal of Personality Assessment, summarizes some of the empirical data that support the validity and potential application of the PIC profile scales. In contrast to Achenbach's subjective critique of selective inventory items, review of scales and scale items provides evidence of their temporal stability, and their ability to classify criterion groups and to predict external correlates obtained independently from parents, teachers, and clinicians. It was also noted that Achenbach's concern over potential distortion of PIC descriptions because they are obtained from informants must be tempered by evidence of scale validity obtained despite the possible presence of such distortion, as well as by direct evidence that suggests the independence of scale elevation and informant psychopathology. PMID- 16370658 TI - The Barnum effect in a group setting. AB - The present study expands the typical Barnum effect paradigm by investigating reactions to feedback in a group setting. People initially participated in a group experience (8 per group), and then were given bogus positive or negative feedback (the favorability manipulation) purportedly prepared by either the group leader or another group member (the source status manipulation). The answerability manipulation led participants to believe that they either would or would not have to share their reactions to the feedback with the person who prepared it. Positive feedback was rated as more accurate and accepted more highly than the negative feedback, though no differences in recall of either the positive or negative feedback emerged. Additionally, a pattern of results revealed that the feedback from the group leader generated greater perceived accuracy, acceptance, and recall than did feedback from another group member. Implications for the impact of feedback given by a high status person in group settings are discussed. PMID- 16370659 TI - Comparison of three psychological defense mechanism questionnaires. AB - The Defense Mechanisms Inventory (DMI), the Coping Operations Preference Enquiry (COPE), and the scales developed by Joffe and Naditch (J&N) were compared using multitrait-multimethod logic. Convergent validity was tow as measures of the same defense were not highly correlated across instruments. Similar patterns of correlations occurred within instruments but the J&N scales, and particularly the DMI scales, had high interscale correlations so overall discriminant validity was limited. These results suggest that either the defense constructs or at least some of the defense scales studied lack validity. Another possibility is that the scales are valid, but assess different parameters of defensiveness (e.g., intensity versus breadth of use of mechanisms). Comparisons of the scales' predictive validity should help resolve these points. PMID- 16370669 TI - Measurement of creativity: review and critique. AB - Presently available criteria of creativity are reviewed and classified into ten categories: tests of divergent thinking, attitude and interest inventories, personality inventories, biographical inventories, teacher nominations, peer nominations, supervisor ratings, judgments of products, eminence and self reported creative activities and achievements. These techniques for measuring creativity are then criticized in terms of their reliability, discriminant validity, dimensionality and convergent validity. It is concluded that an inventory of self-reported creative activities and accomplishments is the most defensible technique for selecting creative individuals. PMID- 16370670 TI - Understanding children's art: an analysis of the literature. AB - This article attempts to answer the question: "Why is it that an intuitively obvious means of understanding children has not been documented as such?". This is done through a critical analysis of the literature dealing with the validity of the Draw-A-Person Test (DAP). Studies reviewed are analyzed along three parameters; projective test theory, subject population, and psychodiagnostic labeling. On the basis of this analysis, explanations for inconsistencies in the results of the research and suggestions for organizing future work in this area are submitted. PMID- 16370671 TI - Primary process integration on the Rorschach and achievement in children: a follow-up study. AB - The purpose of the present study was to follow-up children who had participated in a previous study which investigated the relationship between primary process integration and achievement. The major hypothesis of this follow-up study was supported in that Adaptive Regression (Holt's primary process integration measure on the Rorschach) was significantly positively related to reading achievement (r = .52, p < .001) and to overall achievement (r = .49) in third-grade children. These relationships remained significant when IQ was partialed out. Second-grade AR scores were significantly predictive of third-grade reading (r = .29, p < .05). The correlations between third-grade AR scores and achievement are similar in magnitude to second-grade correlations between these variables. This finding points to the consistency of the relationship between primary process integration and achievement over a one-year period. PMID- 16370672 TI - Reliability of the sentence completion test of ego development in a clinical population. AB - Interrater and internal consistency reliability of Loevinger's sentence completion test of ego development is examined within a psychology clinic population consisting of relatively pure types of neurotic, psychotic, and character disordered groups. Reliability study of the instrument suggests it is generalizable across populations, thusly supporting the reasonableness of further research with the test in clinical contexts. PMID- 16370673 TI - Teacher versus parent reports of preschoolers' social competence. AB - The social competence of children aged three through five was assessed using the Preschool Behavior Questionnaire. Two questions were addressed: First, were the responses of mothers and preschool teachers comparable for any particular child? Second, did the factor structure of the preschool teacher responses replicate those reported by the authors of the instrument? The results showed that the mothers' response patterns, as reflected in the factor structure, proved to differ from those of the preschool teachers. However, the teachers in the Seattle sample did replicate the previously reported factor structures of preschool teachers elsewhere. In addition, it was found that the suggested scoring criteria when applied to the study population produced an unacceptably high number of children in the "deviant" classification. The results were discussed in terms of the implications for using this instrument in general screening programs. PMID- 16370674 TI - A logical learning theory explanation of why personality scales predict behavior. AB - An explanation of why personality scales predict is drawn from the tenets of logical learning theory (Rychlak, 1977). This theory holds that behavior is not only responsive in nature, but also telosponsive, i.e., enacted intentionally for the sake of premises. Personality scales tap the subject's premises concerning some aspect of behavior, the meanings of which are then extended in behavior telosponsively so that a prediction to some criterion performance becomes possible. The subject in effect creates the behavior based on his or her premises. An important telosponse inhuman learning is that of affective assessment, which is operationalized as reinforcement value (like-dislike). Two experiments establish the role of reinforcement value in scale measurement and prediction. The first demonstrates that subjects score higher on personality dimensions which they like very much than on dimensions which they greatly dislike. The second experiment then establishes that a personality dimension which a subject both likes and scores highly on is more predictive to an independently assessed manifestation of this personality characteristic than is a comparable dimension which is disliked. PMID- 16370675 TI - A comprehensive and indexed bibliography of the interpersonal check list. AB - The LaForge-Suczek Interpersonal Check List has been widely used in psychological and social-psychological research, in the evaluation of psychotherapy outcome, and as a component of psychological assessment batteries. Although there is a substantial body of literature on the ICL, access to this literature has been quite limited. In working on the present bibliography, it was found that Psychological Abstracts and computer searches failed to yield sufficient coverage of the ICL literature. It is hoped that this comprehensive, indexed bibliography will make it easier for the interested reader to gain an understanding of the development, application and usefulness of the ICL. PMID- 16370676 TI - The fakability of the personal orientation dimensions: evidence for a lie profile. AB - The resistance of the Personal Orientation Dimensions (POD) to deliberate attempts at creating a favorable impression was investigated. Two groups of college students were used: (a) 21 students completed the POD under standard instructions, then received information about actualization, and completed the POD again under instructions to Create a very good impression; (b) 21 students completed the POD under standard instructions, then received information not related to actualization, and completed the POD again under instructions to create a very good impression. An analyses indicated that providing students with information about actualization did not facilitate their ability to create a favorable impression on the POD. The POD profiles produced by both groups changed significantly, and in a similar direction from pretest to posttest. Evidence is presented suggesting the existence of a "lie profile" which distinguishes deliberate attempts to create a favorable impression on the POD. These findings provide support for the resistance of the POD to deliberate attempts at creating a favorable impression. PMID- 16370677 TI - Reliability of the 16PF questionnaire for security guard applicants. AB - The Sixteen Personality Factor Questionnaire (16PF) was administered to a group of security guard applicants to determine its retest reliability after a moderate (9-month) interval. All of the Pearson correlations for specific test scales were lower than the reliabilities reported by the test's authors. These data raise questions as to the usefulness of the 16PF for the prediction of security guard behavior. PMID- 16370689 TI - The Rorschach: does it have a future? AB - Recent evidence suggests that in spite of an increasingly critical attitude by many clinicians towards the use of projective techniques, the use of these techniques has not faltered over the years. Insofar as the Rorschach continues to be the most popular projective instrument, its scientific status is critically examined in the present paper. The problems inherent in examining an instrument which has engendered a number of quite distinct scoring systems are noted, and the conclusion is advanced that notwithstanding such problems there seems little likelihood that the Rorschach will experience any significantly diminished popularity in the future. PMID- 16370690 TI - Barrier score and the values ascribed to selected individuals by high school students. AB - The Barrier score measure, derived from responses to Rorschach stimuli, was related to the values held by grade 9 high school students. The Barrier score is reflected in a psychological barrier which surrounds the individual and separates him from his environment. High Barrier individuals have a firmer base of psychological operations than their low Barrier counterparts. Groups of high and low Barrier students rated a series of four person stimuli judged to be representative of the outlook of high and low Barrier persons. Ratings were made using a common set of values scales. Multivariate group differences were observed for the stimuli: Sportsman (p = .01), Leader of a Country (p = .03), and Astronomer or Scientist (p = .03), These differences were in terms of derived discriminant functions. Comment is offered on the need for refined techniques for identifying person stimuli compatible with the high Barrier and low Barrier image. PMID- 16370691 TI - A projective measure for the concept of locus of control. AB - The construction and validation of a projective type I-E Scale was devised to measure locus of control. This scale was designed specially to suit the age and culture of a population of school-going children in Sri Lanka. Results indicate that the general feeling of being in control of one's circumstances rather than that of being controlled by outside factors is related to the attitudes one brings to bear on academic achievement and on other achievement related situations. In addition, there was an overall tendency among these subjects towards greater internality. This was interpreted to support the concept of the moral value and the efficacy of effort and hard work. PMID- 16370692 TI - Assessment of private and public self-consciousness: a Dutch replication. AB - In the present article a replication of a study by Fenigstein, Scheier and Buss (1975), who constructed a Self-Consciousness Scale (SCS), is reported. The SCS was translated into Dutch, and its properties and its relation to Locus of Control was investigated. The norms and reliability coefficients presented are consistent with prior research. Also the results of a factor analysis that yielded the three factors: Private, Public Self-consciousness, and Social Anxiety are consistent with these findings. However, Private Self-consciousness did not correlate significantly with Locus of Control. PMID- 16370693 TI - Authoritarianism, dominance and assertiveness. AB - It is shown that there are definitions of the three constructs of authoritarianism, dominance and assertiveness which read very similarly; so much so that no distinction is immediately evident. It is proposed that authoritarianism might be conceived as aggressive dominance and at least some types of assertiveness as nonaggressive dominance. A new scale of Dominance suitable for general population use was produced, and compared with the existing Ray (1976) behavior inventory of authoritarianism. Both scales showed highly significant correlations with peer rated dominance and submission (the latter being negative in sign) but only the authoritarianism scale showed significant correlations with rated aggressiveness and rigidity. It was concluded that the new definitions could be operationalized into valid scales. PMID- 16370694 TI - Dimensionality and reliability of the Rotter I-E locus of control scale. AB - The dimensionality and reliability of Rotter's internal-external control scale was examined using Australian students. The results imply that the scale is not additive but multidimensional. Two distinct largely independent factors relating to personal and political control beliefs were extracted. General control concerns the belief that a person has mastery over his own life while political control includes items which measure the extent to which an individual is capable. of having an influence on political institutions or the course of world affairs. On retesting, 8 of the 9 General Control items loaded on the same first factor. All 5 of the Political Control items loaded on the second factor. Test retest reliability of the I-E scale was .61. These results suggest that the Rotter Scale itself, and its two factor structure, are stable over a considerable period of time. PMID- 16370695 TI - A reanalysis of two scales of tolerance of ambiguity. AB - Important reviews of the tolerance of ambiguity literature have shown that the concept has become overextended and its elements remain unsupported by the confused data collected, and scales with inadequate psychometric performance have been used. In the present study 631 subjects responded to Budner's (1962) test of Intolerance of Ambiguity and MacDonald's (1970) revision of Rydell and Rosen's (1966) Tolerance of Ambiguity Scale. These scales were chosen because they seemed somewhat less suspect than others. Tests of other personality dimensions which the literature reviews suggested were likely correlates of intolerance of ambiguity were also included. Initial results were uneven and inconclusive. After using item analysis to reduce the tests, more logical and consistent results were obtained. These help to clarify the concept, differentiating sharply between its supposed primary and secondary elements. Three studies in which the reduced tests were used with desirable results are reported. PMID- 16370696 TI - Appraisal of lone parenthood after marital breakdown. AB - In order to understand better the experience of becoming a lone parent after marital breakdown a taxonomy of positive as well as negative appraisals and possible coping strategies was constructed. Experiential structures elicited through multidimensional scaling were used to form the basis of the Lone Parenthood Scales. The Scales, with demonstrated reliabilities and construct validity, were then administered to a sample of lone parents who were found to experience concern over their children's limited family life but relief at their separation and hope of becoming closer to their children. They saw themselves as coping with problems by direct, positive action. Further uses for the Scales were discussed. PMID- 16370697 TI - Prorating incomplete Wiggins and MacAndrew Scales. AB - A table is provided for prorating the Wiggins Content Scales and MacAndrew Alcoholism Scale from the Minnesota Multiphasic Personality Inventory (MMPI) when the entire form has not been completed. Conversion factors are given in increments of 10 items for 200 to 400 completed questions. Means, standard deviations, and comparisons of the number of scales over 70 were calculated from 236 profiles, using 200, 250, 300, 350, and 400 items. Results indicate that the conversion factors tend to overestimate scale elevations for the Wiggins scale. The MacAndrew scale can be used for clinical purposes with as few as 200 MMPI items completed, but clinical interpretation of prorated Wiggins scales should be done cautiously. PMID- 16370698 TI - A note on the interpretation of the factor pattern of the California Psychological Inventory. AB - The intercorrelation matrix between the scales of the California Psychological Inventory, given in the manual, was factor analyzed. Three factors were extracted and rotated by varimax. The matrix was rescaled, first normalizing the rows, then standardizing the columns to a mean sum of squares of one. Unity was used as a cut-off point for determination of salient loadings. The first two factors were similar to the results reported in a previous Study by Nichols and Schnell. The third factor was interpreted as an independence-dependence dimension. PMID- 16370699 TI - A review of the Personal Orientation Dimensions Inventory. AB - The Personal Orientation Dimensions Inventory is considered to be an extension and refinement of the concepts measured by the Personal Orientation Inventory. Changes include the elimination of item overlap, an increase in the number of scales and items in the inventory as well as a new theoretical perspective. Significant difficulties still appear to exist in the areas of test construction, theoretical justification, and interpretation. Moreover, the new inventory creates new problems concerning cost for scoring, failure to generate significant amounts of research, as well as a potential loss of relatedness to old research. PMID- 16370709 TI - Orthogonal dimensions of individual and group forms of the Rorschach. AB - Most dimensional analyses of traditional Rorschach scoring categories have either failed to control for differences in total R or have done so by introducing a nonstandard mode of administration. Consequently, the results of such studies may be either distorted or of questionable relevance to more typical situations. The present analyses, which compared both group and individual forms of the Rorschach administered to large samples (over 500 each) of male medical students, controlled for differences in total R by using residualized scores from which the linear effects of R had been partialled. The factor-analytic results are expressed in terms of seven easily interpretable factors (including total R), five of which are similar across forms, and simple formulas for deriving norm referenced factor scores from the data of individual test administrations are provided. PMID- 16370710 TI - Two studies of the movement responses in young children: new and highly discrepant norms. AB - The Holtzman Ink Blot Test was administered to 5- to 6-year-old children and analyzed for movement response. The range and frequency of movement response in both an original and replication study are significantly greater than would have been anticipated for this age group based upon available normative data. NO significant differences were found in the frequency of Human Movement and Animal Movement responses. No sex difference among subjects was found in the range and frequency of movement responses produced. These new findings call into question previously reported data as well as some of the theoretical presumptions underlying the production of movement responses by young children. PMID- 16370711 TI - Sexual ambiguity in children's human figure drawings. AB - Four hundred sixty-one children in grades 5-9 drew a person and indicated the sex of their human figure drawing. A significant number (8%) of children were unable to classify their drawings as to sex; the frequency of uncertainty did not vary with grade or sex of child. Inability to determine the sex of one's own drawing is hypothesized to be a conceptual rather than a perceptual problem, and to reflect uncertainty about the essence of sexual identification. PMID- 16370712 TI - The relationship between arousal and positive self-presentation. AB - Investigated the validity of the hypothesis that subjects would experience arousal if they received information suggesting that they were presenting themselves to others in a negative manner. Twenty-seven subjects were randomly assigned to a control, evaluation apprehension, or negative feedback group. Subjects' heart-rate was continuously monitored as subjects' rated a series of photographs. Subjects in the evaluation apprehension group were given a cue as to how to present themselves positively whereas subjects in the negative feedback group were told, following their response to the 15th photograph, that their responses suggested maladjustment. Heart-rate change above a baseline measure revealed that an increase in heart-rate occurred only when subjects were informed that they were transmitting negative information. Such a finding supports a positive self-presentation explanation of subject motives in the psychological experiment. PMID- 16370713 TI - Field-dependence, sensation-seeking, and preference of paintings. AB - College students (122 males and 88 females) completed the Group Embedded Figures Test measure of field-dependence, and Sensation-Seeking Scale, and performed preference ratings on 40 slides of paintings. Factor analysis revealed six hypothesized and one unhypothesized preference dimensions that underlay preference ratings of paintings. It was hypothesized that the cognitive structuring ability associated with field-dependence would result in certain relationships between field-dependence and preference for paintings. Further, it was hypothesized that preference for novel, complex, and dynamic experiences, associated with sensation-seeking, would result in certain relationships between sensation-seeking and preference for paintings. For the full sample, three of five hypothesized relationships between field-dependence and preference for paintings were obtained, while three of four hypothesized relationships were found between sensation-seeking and preference for paintings. PMID- 16370714 TI - Emotional expressivity and sex-role perceptions of repressors and sensitizers. AB - Two studies were conducted examining the contributions of repression sensitization (R-S) status, sex-role perceptions, and sex of the target of expressivity to assess the expression of emotional discomfort by males, in Experiment I verbal and nonverbal measures of emotional expressiveness were obtained for male repressors and sensitizers exposed to neutral or moderately negative arousal situations with sex of the target of the expressiveness controlled. In Experiment 2 repressors were found to be sex-typed masculine and male sensitizers sex-reversed in their sex-role perceptions. The usefulness of R S status and sex, role perceptions to predicting emotional expressivity by males is discussed. PMID- 16370715 TI - A measure of interpersonal orientation: the Linking People Scale. AB - A 15-item Likert scale, the Liking People Scale, was developed to measure that aspect of interpersonal orientation. Liking people is conceived as a personality construct which serves a mediational function in social interaction. Three cumulative studies are reported. The samples include college students in two regions of the country and a random sample of adults from a large metropolitan area. People low on Liking People spend less time with others, are more socially anxious, tend to be introverted, and are less adept at judging the characteristics of other people. The relationship of Liking People to other measures of affiliative characteristics is examined. PMID- 16370716 TI - A free response test of interpersonal effectiveness. AB - Development of the Interpersonal Problem Solving Assessment Technique (IPSAT), College form, is described. Guided by Rotter's Social Learning Theory, problem solving, and assertiveness research, a semi-structured free response format was designed to assess components of interpersonal effectiveness, The instrument yields patterns of self-reported behaviors in six classes of problematic social situations. A detailed manual enabled reliable scoring of the following response categories: Effectiveness, avoidance, appropriateness, dependency and solution productivity. Scores were not materially affected by sex, verbal ability, or social desirability response sets. Correlations with the College Self-Expression Scale, the Edwards Personal Preference Schedule and the Lanyon Psychological Screening Inventory provided initial evidence of validity. Comparison of mean IPSAT scores of 23 psychotherapy clients with those of 78 normative subjects showed that clients report less interpersonal effectiveness and more avoidance than controls. Implications for utility of the IPSAT are discussed. PMID- 16370727 TI - Miss Theodora Alcock 1888-1980. PMID- 16370717 TI - Locus of control and attribution of responsibility for academic performance. AB - This study tested Phares, Wilson, and Klyver's (1971) hypothesis that locus of control differences are unimportant in a situation which provides very explicit cues arousing specific expectancies regarding the locus of blame for poor performance. Two hundred and seventeen undergraduate students (134 men and 83 women) completed Rotter's I-E Scale as part of classwork in midsemester. During the last week of the semester they were routinely administered a questionnaire containing items on course evaluation, instructor evaluation, and expected grade. Expected grade was significantly correlated with ratings on several course evaluation and instructor evaluation items for internals and externals in the male, female, and total samples. However, these correlations were not significantly different for internals and externals in each sample. These results provided strong empirical support for the tested hypothesis. PMID- 16370728 TI - A computerized method for generating the rorschach's structural summary from the sequence of scores. PMID- 16370729 TI - The Rorschach EA-ep variable as related to persistence in a task frustration situation under feedback conditions. AB - Eighty nonpatients, subdivided into four levels based on the Rorschach EA-ep variable, were randomized into two groups and subjected to a frustrating pursuit rotor task. Extrinsic feedback was provided by a noticeable sound that would occur whenever the tracker was off target. Results indicate that where ep is significantly greater than EA a greater persistence occurred even though the extrinsic feedback signaled no substantial improvement in performance. Two of the three elements contributing to the ep, inanimate movement (m) plus the grey-black and shading answers (T, Y, V, C,), and one variable unrelated to ep, white space (S), account for much of the variance in persistence. The findings are discussed in terms of an overload state that interferes with appropriate processing and mediation in a complex situation. PMID- 16370730 TI - Oral dependence and dependent behavior. AB - Eighty undergraduates were asked to solve a series of difficult puzzles, with the instructions that they could ask for help as often as necessary. Half the subjects had an experimenter of the same sex and half of the opposite sex. The subjects were categorized as high or low dependent based on the percentage of oral dependent responses they gave on the Rorschach test. The results indicated that while the male subjects generally performed as predicted (with high oral dependent males asking for help more than low oral dependent males), the opposite results were obtained for females. Additional analyses suggested that what has been called oral dependence consists of at least two factors that relate in different ways to other variables. PMID- 16370731 TI - Multidimensionality of the Nowicki-Strickland locus of control scale for adults. AB - In order to investigate the dimensionality of the Adult Nowicki-Strickland Locus of Control Scale (ANS-IE), the responses of 120 undergraduate psychology majors were subjected to one principal components factor analysis using Kaiser's varimax criterion, Five meaningful factors emerged which accounted or 80.6% of the total variance. Results were discussed in terms of their similarity to other factor analyses using the children's version of this scale and to a factor analysis of the ANS-IE (chandler & Dugovics, 1977) which used a modified response format. PMID- 16370732 TI - The Narcissistic Personality Inventory: alternative form reliability and further evidence of construct validity. AB - The aim of this study was twofold: (a)to measure the alternate form reliability of the Narcissistic Personality Inventory, and (b)to determine its construct validity by correlating it with the four scales of the Eysenck Personality Questionnaire (EPQ). The alternate form reliability was .72. The Extraversion and Psychoticism scales of the EPQ were positively and significantly correlated with the narcissism measure, and the Lie scale showed a significant negative correlation. The Neuroticism scale showed a nonsignificant relationship with narcissism. In addition, the combined Extraversion and Psychoticism scales produced[ a Multiple R with the narcissism measure that accounted for significantly more of the variance in narcissism than did either measure alone. PMID- 16370733 TI - Family environment and its relation to adolescent personality factors. AB - Investigated the relationship between family social climate characteristics and adolescent personality functioning. The High School Personality Questionnaire (HSPQ) was administered to 80 high school students. These students and their parents also completed the Family Environment Scale (FES). Results of a stepwise multiple regression analysis indicated that one or more HSPQ scales had significant associations with each FES scale. Significant variance in child behavior was attributed to family social system functioning; however, no single family variable accounted for a major portion of the variance to the exclusion of other factors. It was concluded that child behavior varies with total system functioning, more than with separate system factors. PMID- 16370746 TI - Rorschach movement and color responses and MMPI social extraversion and thinking introversion personality types. AB - Results of an analysis of MMPI profiles and Rorschach protocols scored by the Exner method for 100 psychiatric patients provided partial support for Rorschach's hypotheses that perceptions of color (FC, CF, and C) and movement (M) tap separate personality dimensions. First, the proportion of M responses on a protocol correlated significantly (p < .01) to thinking introversion as defined in terms of a subject's MMPI profile similarity to a prototypic MMPI profile with peak PtSc elevations. Second, the proportion of FC responses correlated negatively (p < .01) to social extraversion defined in terms of MMPI profile similarity to a prototypic MMPI profile with peak PdMa elevations. Hence, use of FC signified a socially reserved, controlled behavior. The findings regarding CF and C were inconclusive. Third, color and movement responses did not correlate significantly with somatization defined in terms of similarity to a prototypic MMPI profile with peak Hs, D, Hy elevations. Finally, the nonsignificant correlations of FC with M and with MMPI thinking introversion plus the nonsignificant correlation of M with color responses and MMPI social extroversion further supported the hypotheses that movement and color responses tap essentially unique personality dimensions. PMID- 16370747 TI - The relationship between the Rorschach whole response and level of cognitive functioning. AB - Investigated the relationship between children's level of cognitive development and their production of whole responses (W) on the Rorschach, Both quantitative and qualitative variations in W were examined, and level of cognitive functioning was assessed by Piagetian techniques. The findings indicate that there is a significant and positive relationship between children's stage of cognitive development within the Piagetian framework and the number and complexity of whole responses they produce on the Rorschach. The strength of this relationship appears to be significantly influenced by the stimulus complexity of the inkblots, and the findings suggest that production of highly articulated W on the more complex broken blots requires the abstracting and synthesizing abilities associated with formal operational thought. PMID- 16370748 TI - Orality and latency of volunteering to serve as experimental subjects. AB - A comparison was made of the oral content of Rorschach scores of male Introductory Psychology students who signed up for experiments in the first half of the semester with those who did not sign up until the second half of the semester. Those who participated early in the semester saw more oral images than those who waited until late in the semester to participate (p = .02). The implications of this finding are discussed. PMID- 16370749 TI - A projective analysis of guilt: the Holtzman Inkblot Technique. AB - The group version of the Holtzman Inkblot Technique (HIT) and the three subscales of the Mosher Guilt Scales (Sex Guilt, Hostility Guilt, and Morality-Conscience Guilt) were administered to 44 undergraduate college females in an effort to determine whether or not high-guilt subjects would demonstrate response inhibition on the HIT. The results indicated response inhibition on the part of high-guilt subjects relative to the HIT Human, Movement, and Integration variables. It was concluded that high-guilt subjects responded to the HIT in a style that was less creative and more perceptually immature than low-guilt subjects. PMID- 16370750 TI - The effects of stem length and directions on sentence completion test responses. AB - In an investigation of the effects of stem structure and direction set on sentence completion responses, 160 undergraduate males were randomly assigned to one of four conditions: Rotter ISB with "feelings" instructions, Rotter ISB with "speed" instructions, Sacks SCT with "feelings" instructions, Sacks SCT with "speed" instructions. Eight clinical judges derived hypotheses from the completed protocols; two judges evaluated each protocol. Interjudge reliability was measured by Pearson product-moment correlations and percents of agreement. There was a significant effect for amount of stem structure; the structured Sacks SCT stems yielded more clinical hypotheses. Structured stems also elicited significantly more feeling words. No significant effect was found for instructional set, nor was there a significant relationship between stem structure or instruction set and the numbers of words in completions. PMID- 16370751 TI - Use of local norms to improve configural reproducibility of an MMPI short form. AB - Short forms of the MMPI have been criticized due to their inability to adequately reproduce the profiles of the full length version of the MMPI Oftentimes these short forms are used with subject populations dissimilar to the population upon which the short form was originally standardized. This study evaluates the effect of using local norms with one of these short forms (the FAM) upon the concordance rates of profile code-types with the original full-length MMPI profile. Single point, first 2 high-point and first 3 high-point code-types were found to have higher concordance rates when using local norms. Use of local norms for short MMPI forms may compensate, in part, for information lost in the shortening. PMID- 16370752 TI - Adaptive regression and types of cognitive flexibility. AB - The relationship between adaptive regression and cognitive flexibility was explored in a group of 42 college students. Adaptive regression was measured with the Holt scoring system for the Rorschach. One type of cognitive flexibility was measured by Guilford's tests of divergent production (Word Fluency, Associational Fluency, and Alternate Uses) and a second type by Mednick's Remote Associates Test. Adaptive regression was found to be significantly positively related to the Remote Associates Test in males but not in females. Adaptive regression was found to be unrelated to divergent production tests. These relationships were not found to be mediated by Rorschach productivity or verbal intelligence. A two-stage process was proposed to account for the cognitive operations underlying adaptive regression and the Remote Associates Test. The first stage is seen as a generative (primary process) stage, while the second stage is seen as an evaluative (secondary process) stage. Possible reasons for sex differences were also discussed. PMID- 16370753 TI - Methodological issues in the quantitative analysis of dream content. AB - Reviews measurement approaches applied to dream reports and proposes some methodological cautions that may result in more effective utilization of the dream report in research and assessment. Standardization of dream collection techniques and controls for variability in the length of dream reports are suggested. Further investigation of the intraindividual consistency of dream report measures is seen as necessary for a discussion of their validity. Though there are indications that a variety of dream report measures are potentially valid for psychological assessment, measures based on general dream report characteristics show greater promise than specific content analysis. Research on the differences between normal and disturbed subjects highlights the potential, and the problems, inherent in the use of dream reports as tools of psychological inquiry. PMID- 16370754 TI - Alternate forms of the matching familiar figures test: a critical note. AB - Criticism is leveled at attempts to develop alternative forms of the Matching Familiar Figures test when the literature suggests that what is required is a more reliable test, not simply an alternate version. PMID- 16370755 TI - Validation through self and other ratings on dimensions of six nonstressful multi scaled personality instruments. AB - Six nonstressful personality instruments were concurrently validated using respondent and close friend ratings on specific scale dimensions. Tests were administered to 203 predominantly Caucasian college students. Prior to testing, the individual and a close friend through self-ratings and ratings of the respondents had estimated the strengths of the tested personality Variables on a seven-point scale. The definitions of the various personality dimensions were taken from publisher's manuals. An estimated whole test validity was obtained by an average of the individual scale validities using a conversion of Pearson's r to z'. Significant differences among and between subtest validities were found by ANOVA. All "self-rating" validities were significant (.001), with two of these self-rating validities significantly different (.01) than the other four tests. Only two "other-rating" validities were significant. All six tests appeared to be valid for college population use From preliminary analysis, counselees could rate themselves accurately on most test measures. In some cases self-ratings might be used in lieu of giving the test. PMID- 16370756 TI - Effects of sex and type of humor on humor appreciation. AB - Examined humor appreciation of cartoons as a function of sex of subject and type of humor. Four broad types of humor were presented: sexual-exploitative, sexual nonexploitative, nonsexual-hostile, and nonsexual-nonhostile. Sexual nonexploitative humor was rated as funnier than the other three types but nonsexual-nonhostile humor was given a more positive overall rating than the other types. Relative to males, females gave greater ratings of hostility to the cartoons and rated them less positively. Correlational analyses further suggested that females were not as affected as males by variations in sexuality, exploitation, and hostility. For males, greater ratings of sexuality were associated with greater funniness ratings while the reverse was true for greater ratings of hostility. Possible interpretations of these data were discussed. PMID- 16370767 TI - Introduction to the special series--personality assessment instruments: current status and future directions. PMID- 16370768 TI - Current status of the Rorschach Inkblot Method. AB - The current scientific, clinical, and professional status of the Rorschach Inkblot Method (RIM) is reviewed with respect to its psychometric properties, the applied purposes it can be expected to serve, the extent of its use, and the nature of prevailing attitudes toward it. Available evidence indicates that the RIM is a psychometrically sound measuring instrument that provides valid assessments of personality characteristics and can facilitate differential diagnosis and treatment planning and evaluation. The RIM continues as in the past to be widely used by both clinicians and researchers. However, the esteem in which it is held by practitioners, who are generally agreed that clinical psychologists should be competent in Rorschach assessment, is not universally shared by academicians, many of whom presently question the future place of Rorschach training in graduate education. PMID- 16370769 TI - Current status of MMPI-2 research: a methodologic overview. AB - MMPI-2 research since 1990 has been reviewed to assess whether Butcher and Tellegen's (1978) concerns and suggestions about MMPI research were being followed. Guidelines are provided for when the MMPI-2 is appropriate to administer, how to describe the sample used, assessing validity of the profile, what scores to analyze, and how to report the results. Suggestions also are presented for research within several current areas of debate in the MMPI-2: codetype comparability between the MMPI and MMPI-2, incremental validity of new or existing scales, obvious and subtle subscales, emphasis on item content, development of new scales, and correcting profiles for specific medical and physical conditions. PMID- 16370770 TI - The future of Rorschach in personality assessment. AB - This article considers the Rorschach and its prospects for the next century in the context of two broad issues in psychology, as well as some test specific challenges confronting those interested in the Rorschach and its applications. The broad issues are the matters of personality as a topic of study in psychology, and the relation of findings about personality to treatment planning. The specific issues concern the definition of the Rorschach, research about the test stimuli, and the expansion of information concerning the interpretive usefulness of some structural variables that are poorly conceptualized or for which interpretation may be overly generalized. Some recommendations concerning collaborative research efforts are also offered. PMID- 16370771 TI - Whither goest our redoubtable mentor, the MMPI/MMPI-2? AB - Four areas of MMPI use and development toward the year 2000 and beyond are discussed. First, although the MMPI-2 booklet is a clear improvement, we will continue to need to use both profiles, the MMPI-2 for normative purposes and the MMPI profile for pattern interpretation. Applying MMPI expectations to MMPI-2 profiles is a violation of Meehl's basic actuarial prediction concepts. Secondly, as psychologists begin prescribing medications, we may be able to substantially refine drug choices. Thirdly, the measurement of socioeconomic status levels appears to be the major, missing moderator variable in MMPI/MMPI-2 interpretation. Lastly, an effective and positive MMPI/MMPI-2 feedback paradigm is discussed that fits well within the managed care context. PMID- 16370772 TI - The MCMI-III: present and future directions. AB - Both the original Millon Clinical Multiaxial Inventory (MCMI-I; Millon, 1977) and the Millon Clinical Multiaxial Inventory-II (MCMI-II; Millon, 1987) were refined and strengthened on a regular basis by both theoretic logic and research data. This aspiration has continued. The new Millon Clinical Multiaxial Inventory-III (MCMI-III; Millon, 1994) has been further coordinated with the most recent official diagnostic schema, the Diagnostic and Statistical Manual of Mental Disorders (4th ed., [DSM-IV]; American Psychiatric Association [APA], 1994) in an even more explicit way than before. Although the publication of the first version of the MCMI preceded the publication of the DSM-IV, its author played a major role in formulating the official manual's personality disorders, contributing thereby to their conceptual correspondence. The DSM-III-R (APA, 1987) was subsequently published in the same year as the MCMI-II; the inventory was modified in its final stages to make it as consonant as possible with the conceptual changes introduced in the then forthcoming official classification. The present version of the MCMI, the MCMI-III, strengthens these correspondences further by drawing on many of the diagnostic criteria of the DSM-IV to serve as the basis for drafting the inventory's items. This article reports on a select set of theoretical and empirical developments that are being carefully weighed for possible inclusion in future MCMIs, or as a guide in the refinement process of future MCMIs. PMID- 16370773 TI - Research validity scales for the NEO-PI-R: development and initial validation. AB - The purpose of this investigation was to develop a set of research validity scales for use with the NEO Personality Inventory-Revised (NEO-PI-R; Costa & McCrae, 1992b). In study 1 we used the existing NEO-PI-R item pool to select items for three validity scales: positive presentation management, negative presentation management, and inconsistency. Several iterative item and scale analyses were conducted, using multiple criteria for item selection. These analyses resulted in 10-item scales. In Study 2, the internal consistency, interscale relationships, and normative characteristics were examined in a separate sample of working adults. In Study 3, the validity of the scales was examined by contrasting five sets of NEO-PI-R protocols: from a separate sample of working adults, from a sample of 100 NEO-PI-R protocols with randomly produced responses, and from three samples of undergraduates completing the NEO-PI-R under different instructional sets. Analyses revealed that both the research validity scales and the NEO-PI-R domain scales were sensitive to group differences. issues relating to the appropriate use of the scales are discussed. PMID- 16370774 TI - Detection of random responding on the MMPI-A. AB - We examined random responding on the MMPI-A in 106 adolescents from the general population. Participants were asked to report on the frequency, location, and reasons for any random responses occurring during a standard administration of the MMPI-A. Relationships between self-reported random responding and validity indices (F1, F2, F, and Variable Response Inconsistency [VRIN] scale) were examined. In addition, each participant was randomly assigned to 1 of 5 groups, with each group completing an assigned portion (0, 25%, 50%, 75%, or 100%) of an MMPI-A answer sheet without access to the test booklet, and the utility of the validity scales in discriminating standard protocols from all or partially random protocols was investigated. Most adolescents acknowledged one or more random responses correlated significantly with F but not VRIN. Validity scales were sensitive to all or partially random protocols, and produced high classification rates when discriminating among groups. PMID- 16370775 TI - Personality and depression: a validation study of the Depressive Experience Questionnaire. AB - This study investigated the validity of Blatt's model of depression as indicated by his operational measure of its constructs via the Depressive Experiences Questionnaire (DEQ; Blatt, D'Afflitti, & Quinlan, 1976). Hypothesized relations between the two relevant scales of the DEQ and Tellegen's (1982) Multidimensional Personality Questionnaire, (MPQ) were examined, Participants consisted of 195 women, including 67 hospitalized unipolar depressives, 77 never-hospitalized unipolar depressives, and 51 nonpsychiatric controls. Overall, the results partially supported the validity of the DEQ even though all participants were women and prior studies have indicated the DEQ's greater discriminative validity for men than for women. However, several of the most strongly predicted relations, such as between DEQ Self-Criticism and MPQ Achievement were not confirmed. Coherent, significant relations between scales of the two measures remained after partialling out the effects of severity of depression. PMID- 16370778 TI - SPECIAL SERIES: continuing issues in personality assessment introduction. PMID- 16370779 TI - Assessment of the five-factor model of personality. AB - The five-factor model (FFM) of personality is obtaining construct validation, recognition, and practical consideration across a broad domain of fields, including clinical psychology, industrial-organizational psychology, and health psychology. As a result, an array of instruments have been developed and existing instruments are being modified to assess the FFM. In this article, we present an overview and critique of five such instruments (the Goldberg Big Five Markers, the revised NEO Personality Inventory, the Interpersonal Adjective Scales-Big Five, the Personality Psychopathology-Five, and the Hogan Personality Inventory), focusing in particular on their representation of the lexical FFM and their practical application. PMID- 16370780 TI - Dimensions of deception in personality assessment: the example of the MMPI-2. AB - The topic of deception in personality assessment is discussed along a number of dimensions relevant to clinical practice. The dimensions described are consistency versus accuracy of item endorsement, simulation versus dissimulation, genetic versus specific deception, crude versus sophisticated deception, intentional versus nonintentional deception, self-deception versus impression management, and selectivity versus inclusiveness, as these may be encountered using the revised version of the Minnesota Multiphasic Personality Inventory. The emphasis is placed on deceptive strategies as operations as distinct from the traditional categories of response style such as social desirability. Directions for future research are indicated. PMID- 16370781 TI - Measurement and mismeasurement of mood: recurrent and emergent issues. AB - The affective explosion in psychology has led to tremendous advances in mood measurement. Mood ratings reflect a hierarchical structure consisting of two broad dimensions-Positive Affect and Negative Affect-and multiple specific states. Brief scales have been developed that reliably assess Positive and Negative Affect across different populations and time frames, in both between- and within-subject data. We examine controversies related to (a) the content of these higher order scales and (b) the independence of Positive and Negative Affect. Regarding the latter, we show that Positive and Negative Affect scales remain largely independent across a wide range of conditions, even after controlling for random and systematic error. Finally, there remains little consensus regarding the lower order structure of affect. This lack of a compelling taxonomy has substantially slowed progress in assessing mood at the specific affect level. PMID- 16370782 TI - On the integration of personality assessment methods: the Rorschach and MMPI. AB - Despite being the most studied and used personality assessment tools, data from the Rorschach and MMPI generally disagree (Archer & Krishnamurthy, 1993a, 1993b). Independence is proposed to result from at least 3 factors: (a) the methods tap unique levels of personality, (b) personality has a complex organization, and (c) response styles generate considerable method variance that must be considered in nomothetic research. These ideas led to 5 hypotheses, each of which received support. Rorschach and MMPI response styles are uncorrelated, although response styles are quite consistent within a method family. MMPI-2 and Rorschach constructs of dysphoria, psychosis, or wariness are uncorrelated when response styles are ignored. However, robust convergent validity is evident when patients have similar response styles on each method (e.g., for dysphoria, M r = .59) and dysphoria is expressed in opposing ways on each method when response styles are discordant (i.e., M r = -.54). Data from the latter analyses were correlated with genuine clinical phenomena and implications were discussed for clinical practice and research. PMID- 16370783 TI - Of science, meta-science, and clinical practice: the generalization of a generalization to a particular. AB - Although science is the most powerful method for advancing knowledge and scientific and statistical formalisms are generally much sounder than impressionistic judgment, advanced methodological training (as it is presently comprised) is often of little direct use to practicing clinicians. Practitioners should be able to identify grossly deficient knowledge claims and clear scientific winners, but beyond this, the trick usually is to determine which scientific authority or authorities on whom to depend, a decision task that is ultimately to be tackled through the type of meta-scientific studies that Faust and Meehl have proposed. In applying scientific knowledge, the clinician usually must choose between the generalization of a scientific generalization to a particular versus the application of a clinical generalization to a particular. Determining the relative power of scientific generalizations developed outside, versus clinical generalizations developed within the setting of application, can be very difficult, and there are few established guides. A more advanced science of generalization, another meta-scientific problem, would allow sounder predictions about application to new domains and could greatly aid the practitioner. PMID- 16370784 TI - Reducing MMPI-2 defensiveness: the effect of specialized instructions on retest validity in a job applicant sample. AB - The MMPI-2 is often used for screening job applicants when public safety or security are at risk. Inherent in such applications is concern for profile validity and test defensiveness. In this study, we examine the impact of revised instructions on profile validity for a group of job applicants who initially produced invalid profiles. Participants were 271 male applicants for airline pilot positions. Of these, 72 produced invalid defensive MMPI-2 profiles during preemployment screening. The MMPI-2 was readministered to these applicants with instructions informing them of validity scales and instructing them to respond in a more open, honest manner. Comparisons were made between valid and invalid profiles for initial administrations and between valid and invalid profiles at readministration. Some clinical scales were more elevated for valid, nondefensive profiles. Most content scales showed more elevation for valid profiles, and 12% of the applicants who were retested produced significant elevations (T>or=65) on the content scales. Profiles were similar to those produced by employed pilots of a previous study. PMID- 16370785 TI - Effects of information about validity scales on underreporting of symptoms on the personality assessment inventory. AB - We investigated the effects of information about validity scales on the Personality Assessment Inventory (PAI) on underreporting of symptoms in a college student population. A group of participants completing the PAI under the standard instructions was compared to two groups of participants who had been instructed and given incentives to underreport symptoms. One of the underreporting groups was given a brief warning about the presence of scales designed to detect respondents attempting to present an unrealistically favorable impression. Results suggested that underreporting scales on the PAI are effective in discriminating standard profiles from those completed by uncoached underreporting participants but are less effective in discriminating standard profiles from those produced by participants who have been warned about the underreporting scales. The findings suggest that coaching may enable some respondents to underreport symptoms without detection. PMID- 16370789 TI - A developmental approach to mentalizing communities: I. A model for social change. AB - A developmental model is proposed applying attachment theory to complex social systems to promote social change. The idea of mentalizing communities is outlined with a proposal for three projects testing the model: ways to reduce bullying and create a peaceful climate in schools, projects to promote compassion in cities by a focus of end-of-life care, and a mentalization-based intervention into parenting style of borderline and substance abusing parents. PMID- 16370790 TI - A developmental approach to mentalizing communities: II. The Peaceful Schools experiment. AB - This paper summarizes a theoretical argument for the use of a mentalization-based approach to the systemic problem of school bullying. The Peaceful Schools Project of the Menninger Department of Psychiatry, Baylor College of Medicine, is an experimental test of this model. Data is presented from a randomized controlled trial of this intervention in nine elementary schools in the Midwest. PMID- 16370791 TI - Defense mechanisms and self-reported violence toward strangers. AB - College men who reported committing violence toward strangers (N = 30) and nonviolent college men (N = 30) completed Thematic Apperception Test cards (TAT) and responded to the Minnesota Multiphasic Personality Inventory-2 (MMPI-2). Defense mechanisms were coded from the TAT responses with the Defense Mechanism Manual (DMM). The men who reported violence toward strangers had higher DMM Denial scores and higher MMPI-2 Antisocial Practices scores than the nonviolent men. Student violence toward strangers seems to reflect immaturity rather than psychopathy. PMID- 16370792 TI - Composition and adaptation in the life of Robert Schumann. AB - The composer Robert Schumann, who suffered from bipolar disorder, utilized his impressive musical and literary talents in attempts to adapt to multiple developmental traumas, separations, and losses as well as to the ongoing ravages of his mood disorder. By analyzing several of his Lieder, the author formulates and describes several defense mechanisms involved in these adaptations. These include identification with the lost object, the use of transitional objects and phenomena, sublimation, denial, minimization, idealization, playfulness, and the employment of healthy obsessive traits. Schumann utilized these adaptive defenses successfully for a brief period, thus coping with a difficult separation from his fiancee, Clara. Ultimately, however, he was unable to experience mature mourning regarding the deaths of several family members. The author, drawing on his extensive experience with treating bipolar individuals in outpatient settings, discusses some of the difficulties that many bipolar patients like Schumann have with mourning, including early developmental vulnerabilities to separation and loss, the disorganizing effects of bipolar mood episodes on cognition and self coherence, the need to camouflage affects reminiscent of bipolar mood episodes, and experiencing these affects and mood episodes as traumatic reoccurrences. By studying the biographies of creative individuals such as Schumann, clinicians can expand their appreciation of their patients' adaptive capacities and thus assist them in restoring a sense of hope and vitality in their lives. PMID- 16370793 TI - Psychoeducation for caregivers of patients with chronic mood disorders. AB - Caregivers of patients with mood disorders report high levels of caregiver burden and poor family functioning. This study assessed the impact of educating caregivers about their symptoms and about community resources. Depressive symptoms were reported by 74% of caregivers, but only 37% of these caregivers accepted a referral and only 6% followed through. Similarly, of those with poor family functioning, 44% were offered a referral, 22% accepted, but only 5% followed through. Overall, 82% reported that the psychoeducational packet was useful. PMID- 16370796 TI - Comparison of the MMPI and MMPI-2 administered to Anglo- and Hispanic-American university students. AB - The MMPI and MMPI-2 were administered to White and Hispanic-American subjects. Multivariate analyses revealed significant differences for three main effects: test form, ethnicity, and gender. Two-way interactions between gender and test form and between gender and ethnicity were both significant, but the other two way interaction (Test Form x Ethnicity) and the third-order interaction (Test Form x Ethnicity x Gender) were not significant. Univariate analyses comparing MMPI and MMPI-2 revealed significant differences on 12 of 13 scales, and comparison of Anglos and Hispanics resulted in significant differences on 4 scales. When Anglos and Hispanics taking the MMPI and MMPI-2 were compared, however, no significant differences on any scales occurred. Results suggest that, for these Hispanic subjects, the MMPI-2 introduces no new or additional differences and may be employed and interpreted essentially the same as the MMPI. PMID- 16370797 TI - Personal problem-solving system for scoring TAT responses: preliminary validity and reliability data. AB - Personal problem solving has emerged as an important construct in the cognitive behavioral literature, yet there is a lack of clinically useful, performance based measures practitioners can use to assess the personal problem-solving skills of their clients. Two studies evaluated the validity and reliability of a scoring system for measuring personal problem-solving processes via the Thematic Apperception Test (TAT; Morgan & Murray, 1935). In Experiment 1, undergraduate students (N = 87) completed two measures of personal problem solving, as well as three TAT cards, which were scored using the Personal Problem-Solving System (PPSS; Renan, 1990). In Experiment 2, an additional group of undergraduates (N = 56) responded to three TAT cards on two separate occasions and also completed a different measure of personal problem solving. Results from both studies supported the use of the PPSS for scoring TAT responses to assess personal problem-solving processes. Suggestions for future research are highlighted. PMID- 16370798 TI - Psychometric properties and confirmatory factor analysis of the UCLA loneliness scale. AB - This study investigated the psychometric properties of the UCLA Loneliness Scale. Test-retest and internal consistency reliability were found to be highly acceptable, and support for the concurrent related validity of the scale was also indicated. Confirmatory factor analysis provided support for the unidimensionality of the scale. Evidence was found that the scale is not multivariate normal and in fact appears to be bimodal. Implications point to the need to reconsider the nature of the loneliness construct and to develop conceptually or theoretically based models for testing. PMID- 16370800 TI - The enduring Rorschach contributions of David Rapaport. AB - David Rapaport's contributions to the Rorschach are highlighted and placed within the context of his significant impact on psychological testing and psychoanalytic theory. After a brief biographical sketch, I examine the essence of Rapaport's lasting Rorschach contributions and summarize some of the strengths and limitations of the Rapaport Method. PMID- 16370802 TI - An empirical study of the reports of APA's peer review panel in the congressional review of the U.S.S. IOWA incident. AB - The U.S. House of Representatives Armed Services Committee (HASC) review of the US. Navy's investigation of the U.S.S. IOWA incident provides a case study of the problems with reconstructive psychological evaluations. The US. Navy attributed the explosion on board the U.S.S. IOWA to the intentional suicidal acts of Gunner's Mate Clayton Hartwig, a conclusion supported primarily by an Equivocal Death Analysis (EDA) conducted by the FBI. Systematic analysis of the reports of the expert panelists who consulted to the NASC reveals that 11 of the 14 panelists were critical of the conclusions reached by the Navy/FBI. There was considerable variability among the judgments of the panelists, who reached consensus only for broad categories. Because the reliability and validity of postmortem psychological reconstructions such as EDA and psychological autopsy have not been established, clinicians are urged to exercise caution in the use of such procedures and to be sensitive to ethical obligations to consumers when using such techniques. PMID- 16370803 TI - Code-type comparability of the MMPI and MMPI-2: analysis of recent findings and criticisms. AB - The introduction of the MMPI-2 has stimulated studies of its comparability with the MMPI. Graham, Timbrook, Ben-Porath, and Butcher (1991) consider the congruence between MMPI-2 and MMPI code types substantial, but Dahlstrom (1992) has questioned their appraisal, criticizing their use of "well-defined" code types. Our own analysis supports Graham and colleagues' (1991) conclusions and provides reasons for favoring narrow code types (such as well- defined code types) over nonrestrictive ones. We also offer a brief historical review of MMPI code typology as background for our recommendation that future MMPI-2 research not be limited to studies of code-type correlates. PMID- 16370804 TI - The use of subtle items in detecting deception. AB - When subjects attempt to fake psychopathology on the MMPI, scores on subtle subscales tend to be lower than those of nonfaking subjects. Our study hypothesized that this paradox comes about because the subtle subscales have no predictive validity, but their face validity for psychopathology is the opposite of the keyed direction for psychopathology. Subjects who attempt to fake psychopathology do so on the basis of item content and thus achieve lower rather than higher scores. Three groups of 80 undergraduates took the MMPI under regular, faking-good, or faking-bad instructions. As expected, faking-bad subjects scored significantly lower than regular subjects on the 100 most subtle items, and this was due to their responses to those. 73 of the items whose face validity was misleading. The results are consistent with other work showing valid uses of subtle items in detecting deception. PMID- 16370805 TI - The Rorschach mutuality of autonomy scale in the assessment of black father absent male children. AB - This study examined the utility of the Rorschach Mutuality of Autonomy Scale (MOA, Urist, 1977) in the assessment of two groups of 40 father-absent Black male children (aged 6 to 12 years): those whose fathers had left voluntarily versus those whose fathers had left involuntarily. Children who had experienced involuntary father-absence reflected less adaptive object-relations attainment status as measured by the MOA, t(38) = 2.37, p = > .02, when compared to children whose fathers had negotiated their departure. Although the Rorschach MOA can be a useful measure of assessing psychological functioning in Black father-absent male children, a multimethod assessment of object-relations status for ethnic minority children is proposed. PMID- 16370806 TI - Are critical items "critical" for the MMPI-A. AB - This article examines one aspect of the potential usefulness of critical items to the: Minnesota Multiphasic Personality Inventory -Adolescent (MMPI-A). Endorsement frequency data are presented on the Koss-Butcher (1973) and the Lachar-Wrobeli (1979) critical items for Minnesota Multiphasic Personality Inventory-2 (MMPI-2) adult normative and clinical samples and for MMPI-A adolescent normative and clinical samples. Adolescents in both normal and clinical samples endorse critical items with a higher frequency than do normal adults. Further, results demonstrated that significant differences were uniformly found between the endorsement frequencies for normative versus clinical subjects for the MMPI-2 samples, whereas similar comparisons for the MMPI-A samples typically showed that adolescents in clinical setting did not endorse critical items more frequently than normal adolescents. These findings indicate that it may be difficult to construct critical item lists for adolescents based on the type of empirical methodology used with adults in which items are selected based on endorsement frequency differences found between comparison group. Beyond the issue of the technical difficulty in creating a critical item list for adolescents, several conceptual concerns are raised regarding the application of critical items to the MMPI-A. It was noted that the concept of "critical items" has not been we11 defined, and both the reliability and validity of critical items may be limited in adolescent populations. PMID- 16370807 TI - Comprehensive system scoring discrepancies on Nazi Rorschachs: a comment. PMID- 16370808 TI - Piotrowski's enduring contributions to the Rorschach: a review of Perceptanalysis. AB - Zygmunt A. Piotrowski (1904-1985), one of the early pioneers with the Rorschach technique, developed a method for Rorschach analysis described in the text, Perceptanalysis. Based on clinical, teaching, research, and supervisory experiences, the author selected three aspects of the text for review for their enduring contributions to clinical personality analysis. The three areas are: (a) the scientific and philosophical framework of the system; (b) the Human Movement response, M; and (c) the shading responses--light shading, c'R, and dark shading, c'R. The reader is also introduced to other works by Piotrowski, including scales for cerebral dysfunction and schizophrenia. PMID- 16370809 TI - Psychological tests don't think: an appreciation of Schafer's Psychoanalytic Interpretation in Rorschach Testing. AB - Roy Schafer 's Psychoanalytic Interpretation in Rorschach Testing has, for nearly 40 years, represented a hallmark of psychodynamically informed psychological assessment. In this article, Schafer's contribution is reviewed in light of recent contributions to the Rorschach literature. Following a summary of the main sections of the work, Schafer's approach to the Rorschach is critically re examined from the perspective of modem assessment practice. Differences between the epistemological bases of psychoanalytic and empirical methods of Rorschach interpretation are highlighted, and the prospects for an integration of these two approaches are considered. It is concluded that Schafer's contribution remains as fresh today as when written 40 years ago. PMID- 16370812 TI - Mexican- and Anglo-American differences on the psychological screening inventory. AB - This study investigated differences between Mexican- and Anglo-American college students on the Psychological Screening Inventory (PSI; Lanyon, 1970, 1973)-a brief personality instrument designed to detect persons who might benefit from more extensive evaluations in mental health settings. Further, this study controlled for the Mexican Americans' acculturation level, age, and socioeconomic status (SES), as these variables have routinely been shown to covary with other psychosocial variables. From data on 107 Mexican-American and 105 Anglo-American college students, we found that Mexican-American subjects had higher PSI scores on the Alienation and Defensiveness subscales and lower scores on the Social Nonconformity and Expression subscales than did Anglo-American subjects. With acculturation covaried, however, these differences were no longer statistically significant. With age and SES covaried, the two ethnic groups differed on Alienation, Expression, and Defensiveness, suggesting that Mexican Americans' performance on the PSI varies as a function of acculturation, age, and SES. Implications of these findings are discussed. PMID- 16370813 TI - The development of the items-easy (Ie) and items-difficult (Id) subscales for the MMPI-A. AB - This research involves the development, validation, and cross-validation of the Items-Easy (Ie) and Items-Difficult (Id) subscales for the Minnesota Multiphasic Personality Inventory-Adolescent (MMPI-A; Butcher et al., 1992). These subscales were designed to assess the degree to which reading comprehension deficits may be responsible for significant elevations of validity Scale F and the standard clinical scales on adolescents' MMPI-A profiles. A difference score, bared on the two 13-item subscales, was created in order to compare subjects' responses to subsets of the more comprehensible (Ie) and less comprehensible (Id) items within the test. Hit rate, sensitivity, specificity, positive predictive power, and negative predictive power were calculated on the basis of simple (Id - Ie) and weighted (3Id - 1Ie) difference scores for the validation (N = 495) and cross validation (N = 264) samples, and for specific high-F profile subsamples. Although some of the indices reflected classification accuracy as high as 95%, none of the indices yielded consistently high results across the various samples and subsamples. It has been concluded that the Ie and Id subscales should be used only for research purposes at this time. PMID- 16370814 TI - Dual diagnoses and projective techniques: commentary on Meloy and Gacono's psychotic (sexual) psychopath. PMID- 16370815 TI - Graphomotor interpretation of the MMPI-2? AB - Although clinicians may attribute various personality features to patients who complete paper-and-pencil instruments (e.g., the MMPI-2) with notable neatness (compulsivity?) or sloppiness (oppositionalism?), such inferences have not been empirically examined. In our investigation, MMPI-2 protocols of 154 psychiatric inpatients (74 female and 80 male, M age = 36.7) were examined. A scoring system was developed to categorize the degree of neatness with which each patient had filled in the MMPI-2 response sheet (interrater Cohen Kappa = 0.86). Degree of neatness was not found to be correlated with clinical or content scales on the MMPI-2, with any Millon Clinical Multiaxial Inventory-II (MCMI-II) scales, or with various Rorschach (Comprehensive System) variables. These results imply that, clinical lore notwithstanding, clinicians should refrain from drawing unwarranted inferences about patients' personality features on the basis of the apparent neatness or sloppiness of patients' protocols. PMID- 16370816 TI - The Bender-Gestalt psychopathology scale: failure to infer validity in a school aged sample. AB - The concurrent validity of Hutt's (1977) empirical psychopathology scale of the Bender Visual-Motor Gestalt Test was evaluated among three groups of younger school-aged children within the 7-to-10- year age range (M = 109.1): adjustment disordered, conduct disordered, and nonclinical controls (n = 40/group). Excellent interscorer reliability was achieved with approximately 2 hr of self study of the scoring manual. Although the conduct-disordered group was significantly more immature on Koppitz' Developmental Scoring System, there were no significant differences among groups on the psychopathology scale when visual motor development was covaried (M = 50.5). This was an unexpected result, given Hutt's initial validation study among school-aged children. The psychopathology scale was not inferred to be valid among adolescents in the only known concurrent validation study. Pending supportive validation research, the psychopathology scale should not be employed in the clinical assessment of younger school-aged children. PMID- 16370819 TI - Further validation of computer-assessed response time to emotionally evocative stimuli. AB - Response time (RT) to stimuli from word association test items to Rorschach cards has long been held to be clinically important; especially when unusually long. Recently, Temple and Geisinger (1990) reported longer RT to personality test items previously rated evocative than to other items rated neutral. Our research replicated both Temple and Geisinger's (1990) methods and findings on 23 college students. Subsequent analyses revealed that this effect was found in 78% of all subjects at MMPI-2 item length. Implications for augmenting test validity are discussed. PMID- 16370820 TI - Issues in the comparison of techniques for eliciting source material in computerized content analysis. AB - Zeldow and McAdams (1993) recently presented artifactual explanations for our data showing dissimilarity between the content of speech elicited by the Thematic Apperception Test (Murray, 1943) and free speech tasks (Schnurr, Rosenberg, & Oxman, 1992). In particular, they alleged that our findings resulted from a lack of psychological meaning in our content categories and in the free speech task. We cite empirical and theoretical support to refute this allegation and provide additional analyses of our data that are consistent with our earlier suggestion that text samples elicited under different conditions may not be interchangeable. PMID- 16370821 TI - Construct validity and content analysis. AB - We clarify points of agreement and disagreement with Schnurr, Rosenberg, and Oxman, as put forth in their article appearing this issue. Speech samples elicited under different conditions may or may not be interchangeable. This is a matter of empirical research, but it is also dependent upon the meaningfulness of the content categories. used in the analysis. We prefer a theory-driven, construct validational approach that employs well validated coding schemes to assess psychologically meaningful concepts. PMID- 16370822 TI - Interpersonal correlates of the guilt inventory. AB - The interpersonal correlates of the Guilt Inventory (Kugler & Jones, 1992), which yields scores for state guilt, trait guilt, and moral standards, were assessed. Data collection addressed three issues including the relationships among Guilt Inventory scores and measures of: (a) interpersonal emotions and traits (e.g., shyness, self-consciousness, anger, etc.); (b) the social support network; and (c) ratings of trait guilt, moral standards, and relevant adjectives by friends and family members. Results indicated that trait and state guilt scores were significantly related to various interpersonal measures, most particularly, depression, anxiety, shyness, and loneliness, but only modestly related to satisfaction with social support. Moral standards were generally unrelated to these variables. Others tended to rate participants higher on trait guilt negatively (e.g., higher on contemptuous, angry, guilty, etc.), and participants higher on moral standards in socially acceptable terms (e.g., higher on prompt, self-reliant, moral, and religious). Results are interpreted in view of the frequently cited distinction between social and nonsocial emotions. PMID- 16370823 TI - The relationship between MCMI subtlety and severity. AB - Wierzbicki (Wierzbicki & Daleiden, 1993; Wierzbicki & Howard, 1992) has suggested that the Subtle-Obvious distinction may be useful for detecting faking on the Millon Clinical Multiaxial Inventory (MCMI; Millon, 1983). This study examined Schretlen's (1990) criticism that, on the MMPI, subtlety is confounded with severity. Nine clinical psychology graduate students rated MCMI items for severity. Severity was significantly correlated (r = -.48) with item subtlety. However, this association was lower than that reported by Schretlen on the MMPI. PMID- 16370824 TI - A Review of MMPI and Rorschach interrelationships in adult samples. AB - The MMPI and Rorschach are consistently ranked among the most widely used psychological assessment instruments across adolescent and adult clinical settings. Although there is an extensive research literature available on each instrument individually, relatively little research attention has been focused on the interrelationships between these measures. This article reviews the literature derived from 37 studies that have reported interrelationships between MMPI and Rorschach variables in adult populations. The results of these studies generally indicate limited or minimal relationships between the MMPI and Rorschach. A number of methodological issues, however, prevent drawing firm conclusions from the literature at this time. Directions for future research are discussed, including consideration of issues related to sources of alpha and beta error and the need for studies examining the incremental validity of combinations of Rorschach and MMPI data in prediction to relevant external criteria. PMID- 16370825 TI - Rorschach oral imagery, attachment style, and interpersonal relatedness. AB - Data on a college sample (n = 198) were used to examine the hypothesized relationship between Rorschach orality and self-reported attachment style. The number of Rorschach Human responses was also examined. High orals were more often classified as insecure than low orals. Within the low-oral group, however, subjects who produced no oral responses were more likely to be classified as insecure than subjects who produced one oral response, 78% versus 35%; p < .005. This implies that there is heterogeneity within the low-oral group. Neither orality nor attachment security was meaningfully related to the number of Human responses provided, but the Human responses provided by insecure (vs. secure) subjects more frequently evidenced themes of struggle. We concluded that (a) there is a subgroup of low orals who are not interpersonally secure but who, instead, are insecure and interpersonally anxious; and (b) unlike the quality of the Human response, the number of Human responses may not reflect preoccupations with themes of dependency, attachment, and interpersonal relatedness. PMID- 16370826 TI - A Rorschach analysis: parallel disturbance in thought and in self/object representation. AB - The relationship between thought process disturbance and the structure of internal object relations was investigated by comparing Rorschach Special Scorings and content variables that reflected developmental levels of object relations. Selected Special Scorings from the Comprehensive System (Exner, 1990) and composite (WSum6) scores were correlated with Urist's (1977) Mutuality of Autonomy Scale (MOA) on the Rorschach protocols of 35 borderlines, 15 schizophrenics, and 19 narcissists. Correlations between composite scores of severe thought process disturbance and severe imbalance in internalized object representations were highly significant, p < .0001. The hypothesis that there is a significant relationship between disturbance in internalized self/object differentiation and thought process disturbance was supported. Implications of this study are discussed as they relate to empirical validation of theoretical constructs concerning ego function deficit and self/object disturbance. PMID- 16370827 TI - The rorschach search for the borderline Holy Grail: an examination of personality structure, personality style, and situation. AB - Much of the research on the use of the Rorschach to diagnose borderline disturbance has sought to discover a specific borderline pattern or Rorschach configuration. Given our increased understanding of borderline functioning representing a level of personality organization (or personality structure), this single pattern approach is excessively simplistic. It fails to consider the complex interaction between personality structure, personality style, and situational variables. An approach to the Rorschach assessment of borderline functioning is presented using a clinical example. The patient was tested at the onset of psychotherapy and then some 4 years into treatment. Results from the two testings were compared using Weiner and Exner's (1991) Rorschach variable clusters for assessing change in psychotherapy. PMID- 16370828 TI - A borderline policeman: AKA, a cop with no COP. AB - Lest the Rorschach partisan engage in splitting and deem one approach to clinical data as all good and the other as all bad, it is urged that atheoretical, nomothetic and theory-saturated, developmentally based, idiographic approaches be integrated to animate, enrich, and organize the mature psychodiagnostician's yield. This approach is illustrated by exploring the borderline personality organization of a 46-year-old police officer fearing fragmentation and loss of control. PMID- 16370829 TI - Training in psychodiagnostic testing in APA-approved PsyD and PhD clinical psychology programs. AB - Fifty-one percent (N = 80) of directors of doctoral PhD and PsyD programs in clinical psychology, accredited by the American Psychological Association (APA), responded to a survey on assessment training. The study was a replication of an earlier survey by Piotrowski and Keller (1984b) on instruction, practices, and attitudes on testing. Our findings indicated that training emphasis in assessment coursework has changed very little over the past decade. Intelligence testing was emphasized in most programs, followed by objective and projective personality assessment. However, training in behavioral assessment techniques was required in only half of the responding programs. There were no significant differences between PhD and PsyD programs concerning required coursework in assessment. Factors that influence the perennial importance of testing in the clinical core curriculum are discussed. PMID- 16370832 TI - A psychometric evaluation of the new NEO-PIR facet scales for agreeableness and conscientiousness. AB - The five-factor model of personality represents one of the more important developments in the area of personality theory and assessment. This empirically derived model consists of the major factors of Neuroticism, Extraversion, Openness to Experience, Agreeableness, and Conscientiousness. Currently there is only one commercially available measure of these dimensions: The NEO Personality Inventory (NEO-PI). The NEO-PI measures each of these global domains as well as more specific facets of Neuroticism, Extraversion, and Openness to Experience. The new revision of the NEO-PI (NEO-PIR) now includes facet scales for Agreeableness and Conscientiousness. The purpose of this article is to provide a psychometric evaluation of these new scales using a sample of working adults and relying on both self-report and observer ratings. The results provide strong support for the reliability and construct validity of these new scales. PMID- 16370833 TI - On the Rorschach M response and monotheism. AB - The Rorschach human movement (M) response has recently been debated (Exner, 1991a, Kramer, 1991a, 1991b) with additional wider implications for Rorschach psychology. The objective of this discourse is twofold: (a) to elaborate on the contributions of perceptanalysis (Piotrowski, 1957) to this debate and (b) to develop the thesis that diversity is desirable for the long-term advancement of the field of Rorschach psychology and personality assessment. These motifs are illustrated with an interpretation of a clinical case example and with rationale from the theoretical literature and implications for interpreting short Rorschach protocols. The conclusion is proffered that some degree of diversification is more beneficial than requiring the adoption of a single Rorschach approach (i.e., monotheism). PMID- 16370834 TI - DIStance-from-the PROtotype (DISPRO) personality assessment for children. AB - We examined a children's version of Broughton's (1990) DISPRO (DIStance-From-the PROtotype), a multidimensional scaling approach to personality assessment. In this system, a score on a given trait dimension is derived from a subject's judgment about his or her similarity to or distance from a prototype - a hypothetical character who acts in trait-consistent or prototypical ways. The DISPRO assessment model employs eight equidistant trait categories from Wiggins's (1979) interpersonal circumplex. In this study, 74 children (M age = 7.3 years) were told stories about four same-sex characters who performed prototypical acts from a single trait category randomly selected from the circumplex. Results show the structure of children's ratings is remarkably similar to that obtained from adults, but children's ratings show a developmental trend. A sample of younger children (M age = 6) provided a unidimensional solution along a "good-bad" or nurturance continuum. Analysis of an older sample (M age = 8.5) revealed use of the two dimensions of dominance and nurturance commonly found in adult solutions. Significant validity coefficients for parent and teacher criterion measures (average r = .40) and adequate test-retest reliability (average r(tt)= .78) were found. Benefits unique to DISPRO, such as its ease of use and standardization of trait stimuli, are discussed. PMID- 16370835 TI - A new test of an old hypothesis: a quantification of sequence in the Rorschach. AB - Rorschach interpretation often assumes that successive responses are not independent of one another but rather that they are part of a series of interconnected events. In this study, methods that have been used to analyze event sequence data were applied to Rorschach protocols. Results from a nonclinical group of 102 university students showed that location scores on successive responses were repeated more frequently than was predicted by chance. There was also a tendency for subjects to make transitions from larger to smaller or more detailed areas of the inkblot on successive responses. In addition, we found that subjects tended to make transitions from more adequate to less adequate use of form, and that the unusual and minus form categories tended to be repeated. A modest association between transition frequencies and individual differences in anxiety, but not between transition frequencies and depression or overall symptomatology, was demonstrated. PMID- 16370836 TI - Some NEO-PI five-factor personality profiles. AB - The aim of this study was to identify any clusters of score profiles to be found in a college sample of 236 subjects administered the five-factor NEO [Neuroticism, Extroversion, Openness] Personality Inventory (Costa & McCrae, 1985). Application of Ward's agglomerative hierarchical procedure to the score profiles disclosed six clusters that were replicated in a K-means partitioning process. The six clusters were then compared by a one-way analysis of variance (ANOVA) with respect to their mean five higher order scores on the Interpersonal Style Inventory (Lorr, 1986). The highly significant F tests provided confirmation of the characteristics of the cluster profiles isolated. PMID- 16370837 TI - Clinical considerations in the conjoint use of the Rorschach and the MMPI. AB - Contemporary clinical and research findings concerning the Rorschach and the Minnesota Multiphasic Personality Inventory (MMPI) indicate that (a) objectivity and subjectivity are relative and not categorical dimensions of these two instruments; (b) apparent contradictions between Rorschach and MMPI results are generative and not invalidating; (c) within limits, false negative findings are not cause for concern in the clinical application of Rorschach and MMPI variables; and (d) differences between the Rorschach and the MMPI in how they are structured and what they measure demonstrate considerable advantage in using them in complementary fashion to support clinical inferences. PMID- 16370838 TI - On the comparison of TAT and free speech techniques in personality assessment. AB - Schnurr, Rosenberg, and Oxman (1992) recently compared the free speech samples and Thematic Apperception Test (TAT) responses of 95 normal adults. They concluded that the two techniques are not interchangeable, and that the TAT, which proved superior in the prediction of individual differences, may be preferable to free speech instructions for eliciting data in content analytic studies. We disagree with both conclusions. Various forms of narrative speech samples may be highly correlated, so long as psychologically meaningful, well validated, and higher order content categories are used. The use of first-order content categories is less likely to contribute to the study of personality. PMID- 16370841 TI - Assessing concrete and abstract thinking with the Draw-a-Person technique. AB - In this study, we examined the Draw-a-Person (DAP) technique as an indicator of concrete/literal and abstract thinking, Subjects (N = 50) included schizophrenic inpatients (n = 25) and college students (n = 25). Fourteen possible indicators of concrete/literal or abstract thinking on the DAP were generated based on theoretical considerations and prior research. Five indicators were eliminated because of their low interrater reliability. The Proverbs Test (Gotham, 1956) was used to establish evidence of concrete/literal and abstract thinking. Scores from the DAP and the Proverbs Test were correlated. Composite scores on the DAP were highly correlated (r = .52, p < .01) with the corresponding Proverbs Test score. In addition, abstract scores on the DAP were correlated (r = .54, p < .01) with a similar Proverbs Test score.. Concrete/literal DAP scores did not correlate significantly (r = .20, p > .05) with concrete/literal Proverb Test scores. Results indicate the potential for using the DAP technique to assess concrete/literal and abstract thinking when a nonverbal test is needed. PMID- 16370842 TI - The influence of situational and interpersonal variables in projective testing. PMID- 16370845 TI - Adlerian-oriented early memory research: what does it tell us? AB - Of the different approaches to interpreting early memories (EMs), Adlerian oriented EM research comparatively has shown a most substantial increase in the past 10 years. I examine all Adlerian-oriented EM research appearing from 1981 through 1990. The majority of the research provides a tentative base of support for Adler's (1931) views about EMs. However, several deficiencies in these investigations are identified (e.g., a lack of pilot studies), and some recommendations are made for future EM studies. PMID- 16370846 TI - The consistency of scores and configural patterns between the MMPI and MMPI-2. AB - This article is an extension of the Ben-Porath and Butcher (1989a) study, investigating the claim that the scores and configural patterns of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) are consistent with those of the Minnesota Multiphasic Personality Inventory (MMPI). College students (200 men and 200 women) were randomly assigned to either the original to original condition (O O), in which they took the MMPI twice, or the original to revised condition (O R), in which they took the MMPI and the MMPI-2. Correlations and configural patterns of the O-O condition were compared to those of the O-R condition. Results suggest some score changes between the MMPI and MMPI-2, particularly on Scales L, 5, and 8 for men and Scales L and 5 for women. PMID- 16370847 TI - Personality and spirituality: comparative TATs of high-achieving Rajneeshees. AB - The Thematic Apperception Test (TAT) was administered to followers of Bhagwan Shree Rajneesh in their commune in central Oregon. The Rajneeshee subjects had been high achievers (with advanced degrees and/or previous incomes greater than 30,000 dollars in 1984 dollars). Later, TATs were administered to persons matched for gender, age, education, and occupation but not involved in new religions. The final matched sample included 24 men and 20 women, half of whom were Rajneeshees. TATs were coded for a variety of formal variables as well as personality ratings. Though the analysis showed no between-group differences on most variables, significant mean differences occurred on ratings of unusual conditions, quest, and narcissism, with Rajneeshees higher than comparison subjects. Gender differences in personality variables were not conspicuous. Factor analysis established a strong productivity element. Card 16 (the blank card) provided the greatest number of differences. PMID- 16370848 TI - The analysis of descriptions of parents: identification of a more differentiated factor structure. AB - The analysis of spontaneous descriptions of parents has proven to be a valuable procedure for assessing an individual's perceptions of caring experiences with their parents. The procedure has been useful in a number of research investigations. Blatt, Chevron, Quinlan, Schaffer, and Wein (1988) revised the scoring procedures in a new manual for this procedure. This study examined aspects of the reliability and validity of this new scoring system. The parental descriptions of 134 female and 79 male students from two universities were scored using the revised manual. An analysis of ratings from the new scoring procedures lead to a differentiation of negative punitive items from positive ambitious items. Three factors and a Length factor emerged; the four factors accounted for 71% of the variance. Three factors-Benevolent, Punitive, and Ambitious-were correlated in meaningful ways with relevant scales of the Semantic Differential, the Zung Self-Rating Depression Scale (Zung, 1972), and the Depressive Experiences Questionnaire (Blatt, D'Afflitti, & Quinlan, 1976). The results provide construct validity for this new factor solution for evaluating the descriptions given by individuals of significant figures in their lives. PMID- 16370849 TI - Statistical power and the Rorschach: 1975-1991. AB - The Rorschach Inkblot Test has been the source of long-standing controversies as to its nature and its psychometric properties. Consistent with behavioral science research in general, the concept of statistical power has been entirely ignored by Rorschach researchers. The concept of power is introduced and discussed, and a power survey of the Rorschach literature published between 1975 and 1991 in the Journal of Personality Assessment, Journal of Consulting and Clinical Psychology, Journal of Abnormal Psychology, Journal of Clinical Psychology, Journal of Personality, Psychological Bulletin, American Journal of Psychiatry, and Journal of Personality and Social Psychology was undertaken. Power was calculated for 2,300 statistical tests in 158 journal articles. Power to detect small, medium, and large effect sizes was .13, .56, and .85, respectively. Similar to the findings in other power surveys conducted on behavioral science research, we concluded that Rorschach research is underpowered to detect the differences under investigation. This undoubtedly contributes to the inconsistency of research findings which has been a source of controversy and criticism over the decades. It appears that research conducted according to the Comprehensive System for the Rorschach is more powerful. Recommendations are offered for improving power and strengthening the design sensitivity of Rorschach research, including increasing sample sizes, use of parametric statistics, reduction of error variance, more accurate reporting of findings, and editorial policies reflecting concern about the magnitude of relationships beyond an exclusive focus on levels of statistical significance. PMID- 16370850 TI - Three loneliness scales: an assessment of their measurement properties. AB - In loneliness research, there has not been a consensus concerning either the factorial structure of most loneliness scales or the factorial structure of each of the proposed dimensions of loneliness. We investigated theoretical and operational definitions, as well as the factorial structure, based on gender of three loneliness scales: the University of California at Los Angeles (UCLA) Loneliness Scale, the Loneliness Deprivation Scale, and the Emotional/Social Loneliness Inventory. Unidimensional, best-fit models of the structures of these scales are presented. PMID- 16370852 TI - Parental idealization and the absence of Rorschach oral imagery. AB - Although the attitudes, behaviors, and pathologies associated with the oral personality have been examined extensively and are well-described (Masling, 1986), the absence of Rorschach oral imagery has not been studied. Based on previous research, we hypothesized that undergraduate subjects who do not provide any oral responses idealize at least one of their parents. This hypothesis was tested by asking subjects to describe their parents (Blatt, Wein, Chevron, & Quilan, 1979). Indeed, subjects who do not report perceiving any Rorschach oral imagery are more likely than subjects who do report oral imagery to engage in parental idealization. The results are discussed in the context of previous research on dependency and attachment. PMID- 16370853 TI - Projection, transitional phenomena, and the Rorschach. AB - This article sheds light on the theoretical foundations of projective testing. Drawing on the psychoanalytic perspective, especially Winnicott's works, it elucidates the nature of projection and transitional relatedness, with particular reference to the Rorschach. This conceptualization of the Rorschach task is compared with Exner's more cognitive-behavioral orientation, which holds that the Rorschach is not a projective test. I suggest that Exner's Comprehensive System could become more truly comprehensive if it were integrated with a more complex understanding of the response process, allowing more importance to the role of stimuli from the inner world to counterbalance its focus on the processing of external stimulation. Clinical and research implications are discussed. PMID- 16370854 TI - The Rorschach's factor structure: a contemporary investigation and historical review. AB - The Rorschach Inkblot Test was factor analyzed to assess for a two-dimensional structure that was expected to reflect the traditional interpretation of many scores, as well as two dimensions that have become a basic paradigm for studying self-reported personality and mood. In my sample (N = 268), Comprehensive System scores had a four-dimensional structure. These factors replicated many earlier findings but were independent of self-report measures. Two of the Rorschach dimensions were strongly defined by response frequency (R) and reflect the powerful impact that R has on determining scores and their intercorrelations. A third dimension was comprised of form-dominant shading determinants and partially corresponded to a hypothesized factor of dysphoria. The fourth dimension was comprised of holistic nonform-dominant color and shading determinants and appears to reflect Shapiro's (1965) description of the hysterical mode of cognition. This study suggests it is necessary to expand our conceptualization of the Rorschach to account for its reproducible factors. PMID- 16370855 TI - Comparability of two-point high-point code patterns from original MMPI norms to MMPI-2 norms for the restandardization sample. AB - Using records from the 1,138 males and 1,462 females in the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) restandardization sample (Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989), two-point high-point code patterns generated from the original norms were compared to the patterns that these subjects obtained from the new norms. Although some code patterns proved to be quite stable across both norms, code comparability was generally lower in this community-based sample than was true for the records from samples of psychiatric patients also reported in Butcher et al. (1989). The sources of differences between the original and the new norms were reviewed, and the implications for profile interpretation based on code patterns were pointed out. The differences arising from the use of the MMPI-2 norms are appreciable; they highlight the need for new empirical data on the correlates of coding patterns based on these norms. PMID- 16370858 TI - Projective techniques in the computer age. PMID- 16370856 TI - Profile clusters for men and women on the personality adjective check list. AB - This study examined the major personality profiles found for men and women on the Personality Adjective Check List (Strack, 1991b). Subjects were 1,058 men and 1,194 women from a number of samples of normal adults (Strack, 1991b). Ward's (1963) agglomerative hierarchical procedure yielded five clusters for men and four for women that were replicated in subsequent K-means nonhierarchical analyses. Correlates of cluster membership obtained from subgroups of subjects on four personality measures provided strong evidence for the classification. PMID- 16370859 TI - Thematic analysis of daily events as a method of personality assessment. AB - I examined how themes in emotionally meaningful daily events can be used as a personality assessment technique. Eighty-eight student participants recorded their most positive and most negative experiences each day for 2 weeks. Common themes were identified in these events by the participants and judges. The convergent validity of the daily events measures was demonstrated through comparisons with a diverse set of personality measures of achievement and affiliation motivation, including a projective test (the Thematic Apperception Test), self-report questionnaires, and peer reports. Results suggest that a thematic analysis of daily life events may be a useful means of assessing the specific daily concerns and stressors experienced by individuals. PMID- 16370862 TI - The problem of R in the Rorschach: the value of varying responses. AB - A subject's freedom to give different numbers of responses to each inkblot has always been part of the Rorschach tradition. However, frequency of response (R) as a variable rather than a constant requires special attention when using the Rorschach psychometrically and when making clinical evaluations based in part or entirely on normative scores. In this article, I discuss the Rorschach tradition; the distinctiveness of the procedure; and the complexity of the technical, methodological, and theoretical problems. issues related to the integration of the empirical and conceptual approaches are discussed. PMID- 16370863 TI - Response frequency problems in the rorschach: clinical and research implications with suggestions for the future. AB - This article focuses on clinical and research problems associated with response frequency (R) being a variable in the Rorschach. Despite the fact that variations in R directly contribute to 50% of the explainable variance among Rorschach raw scores, there is a dearth of empirical evidence to document what R actually measures. Furthermore, in the practical use of the Rorschach's structural data, R is considered to be a nuisance variable that is controlled and not deemed interpretively significant. Given this information, two research agendas are proposed. The first is to more thoroughly determine whether R measures anything of substantial clinical importance. The second is to evaluate systematically the relative merits of making R a constant rather than a variable through use of an R controlled method of Rorschach administration. This strategy would resolve many of the psychometric problems related to R. Introducing greater structure and clearer expectations to the task may also sharpen the Rorschach's ability to assess and predict important aspects of personality. However, significant disadvantages would also result from this change in administration. Both sides of the issue are discussed in some detail. PMID- 16370864 TI - R in Rorschach research: a ghost revisited. AB - As interest in Rorschach research has increased, an old issue-whether differences in R require that other scores be normalized in relation to R when Rorschach data are analyzed-has been resurrected. Most who advocate normalizing or partialing for R cite Cronbach (1949) or Fiske and Baughman (1953) as the basis for their argument. Although Cronbach offered several useful suggestions regarding this problem, the data base to which he referred and that used by Fiske and Baughman allowed for a much greater variation in R than occurs when Rorschach's standard instructions are applied. Intercorrelations between R and most parametric Rorschach variables are not significant among groups of nonpatients, schizophrenics, and depressives. Thus, although the issue of R can be critical when some variables are included in a data matrix, most analyses can proceed without partialing or normalization. When nonparametric variables are included in an analysis, partialing or normalizing is inappropriate because the resulting distributions will be distorted. PMID- 16370865 TI - The problems of R in clinical settings and in research: suggestions for the future. AB - It has been suggested that we control for R in the interpretation of individual Rorschach records. A related, but distinctly different issue concerns controlling for or equating R in research. I suggest we should not control for R in interpreting individual records. There are also some research paradigms in which controlling for R does not seem advisable. In other types of research, I argued that some method of controlling for or equating R appears appropriate. The distributions of many Rorschach factors are markedly non-normal, and in some cases the shapes of the distribution for a particular factor may be different among different populations. In these instances, when attempting to control statistically for R, improper model specification may lead to erroneous findings. Finally, I argue for a theory-based, empirical approach to all future Rorschach research. PMID- 16370866 TI - Equivalence of computerized and standard administration of the Piers-Harris Children's Self-Concept Scale. AB - Two hundred seventy-nine 8- to 17-year-old children and adolescents were randomly assigned to complete paper-and-pencil or computer-administered versions of the Piers-Harris Children's Self-Concept Scale. The equivalence of these two administration modes was assessed by evaluating the comparability of scale means, variances, reliabilities, and validities. Results indicate that, of 27 comparisons, only 1 produced a statistically significant difference. We concluded that the computerized administration of the Piers-Harris scale did not affect its psychometric properties and that paper-and-pencil and computerized modes of testing for this questionnaire may be regarded as equivalent. PMID- 16370867 TI - Comparison of TAT and free speech techniques for eliciting source material in computerized content analysis. AB - We compared the free speech samples and Thematic Apperception Test (TAT) responses of 95 community-residing volunteers by using the General Inquirer content analysis computer program and the Harvard III Psychosociological Dictionary (Stone, Dunphy, Smith, & Ogitvie, 1966). Comparability was assessed by computing mean differences and correlations among techniques. The techniques were evaluated by assessing the ability of data derived from each to predict individual differences in developmental level, gender, depressive symptomatology, and personality. Results show highly discriminable profiles and low reliability among techniques. TAT data were superior in predicting individual, differences. We suggest that structured techniques like the TAT are preferable to standard free speech instructions for eliciting data in content analytic studies and discuss the possibility of computerized content analysis as a method of scoring the TAT. PMID- 16370868 TI - The early memories procedure: a projective test of autobiographical memory, part 2. AB - In Part 1 of this paper, topics germane to the projective assessment of autobiographical memory were discussed. In Part 2, the cognitive-perceptual method will be used to interpret several early memories and a slightly abridged Early Memories Procedure (EMP). PMID- 16370869 TI - Fear of success, fear of failure, and the imposter phenomenon: a factor analytic approach to convergent and discriminant validity. AB - A sample of 104 mid-level managers completed measures of fear of success, fear of failure, and the imposter phenomenon. First- and second-order factor analyses indicated that the instruments tapped two personality domains, lack of self confidence and the competitive neurosis. Results are discussed in terms of the most appropriate use of the instruments employed in the analysis and the need to avoid redundancy in the development and popularization of new psychological constructs. PMID- 16370870 TI - Profile of the membership of the Society for Personality Assessment: comparisons between 1987 and 1990. AB - In this study, we report the findings of a 1990 survey of the membership of the Society for Personality Assessment (SPA) and compare these results with a similar survey conducted in 1987. A four-page questionnaire was sent to approximately 1,800 SPA members; 900 surveys were returned, for a 50% response rate. Information was obtained on professional activities, occupational setting, theoretical orientation, and various demographic characteristics. Clinical practice was reported as the primary professional activity of the vast majority of SPA members; more than 40% of the 1990 sample indicated private practice as their primary job setting, compared to 35% in I987. A larger proportion of SPA members are also now working in private/clinical practice, medical centers, and outpatient community clinics than in 1987. Although a psychodynamic psychoanalytic orientation continues to be the dominant theoretical perspective of SPA members, the proportion of members with a cognitive-behavioral orientation has increased substantially since 1987. We concluded that SPA members are now more involved in clinical practice, and that the SPA has become more ecumenical in theoretical perspective as the membership has more than doubled over the past 3 years. PMID- 16370871 TI - The HIT and the HIT 25: comments and clarifications. PMID- 16370874 TI - Assessing the validity of computer-based test interpretations: rating reliability and individual differences among raters. AB - Three studies were conducted to assess the role of individual differences among raters participating in validation studies for computer-based test interpretations (CBTIs) and to assess the reliability of ratings. Studies centered on two CBTIs for the Sixteen Personality Factor Questionnaire (16PF). The first study involved 54 students who rated the accuracy of the Human Resource Development Report (HRDR) in an experimental context. The second study involved 73 students who rated the HRDR in a nonexperimental context. The third study involved 28 students who rated the Narrative Score Report (NSR) in a nonexperimental context. Results taken together indicate that (a) shrewdness may influence ratings of CBTI accuracy in an experimental context; (b) self sufficiency, emotional stability, dominance, or abstract thought may influence ratings in a nonexperimental context, depending on the specific report involved: (c) test-retest reliabilities for accuracy ratings were .74 for HRDR and .81 for NSR; and (d) test-retest reliabilities for usefulness ratings were .75 for HRDR and .54 for NSR. PMID- 16370875 TI - Workaholism: definition, measurement, and preliminary results. AB - Questionnaires were developed to assess the concept of workaholism, defined in terms of high scores on measures of work involvement and driveness and low scores on a measure of enjoyment of work, and to contrast this profile with work enthusiasm, defined as high work involvement and enjoyment and low driveness. Additional scales were devised to test several predictions about the correlates of workaholism. A test battery including these scales was given in a mail survey to a national sample of male (n = 134) and female (n = 157) social workers with academic positions. The psychometric properties of the scales are described. Cluster analyses for each sex revealed groups who corresponded to the workaholic and work enthusiast profiles as well as several other profiles. As predicted, workaholics were higher than work enthusiasts (among other groups) on measures of perfectionism, nondelegation of responsibility, and job stress. They were also higher on a measure of health complaints. Investigations are being initiated to determine the association of workaholism and other score profiles with objectively diagnosed cardiac disorders and with measures of occupational performance. PMID- 16370876 TI - Social support and psychological well-being in young adults: the multi dimensional support scale. AB - A large sample (N = 483) of community-living adults (M age = 23.6 years) responded to questions about the frequency and adequacy of emotional, practical, and informational support during the past month from three sources-confidants, peers, and supervisors-using the Multi-Dimensional Support Scale. Factor analysis was used to distinguish meaningful regularities in item responses, and the resulting five scales have high internal reliability. Scores correlated significantly with measures of self-esteem, depressive affect, and psychological disturbance according to the General Health Questionnaire. To investigate what contribution support measures could make to the prediction of psychological well being, hierarchical regression analyses were carried out with stressors entered first and support scores entered second. As expected, respondents' psychological well-being was related to their level of life stress; however, the addition of social support measures doubled the explained variance in measures of psychological well-being. The support measure with the strongest association with well-being was the frequency of supportive behaviors toward the respondent by close family and friends. PMID- 16370878 TI - Reminiscence, retirement, and Rorschach responses in old age. PMID- 16370879 TI - On the psychometric properties of the short form of the expanded attributional style questionnaire: a response to Peterson. PMID- 16370883 TI - Special series: the MMPI-2. PMID- 16370884 TI - Comparability of computerized adaptive and conventional testing with the MMPI-2. AB - A computerized adaptive version and the standard version of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) were administered 1 week apart to a sample of 155 college students to assess the comparability of the two versions. The countdown method was used to adaptively administer Scales L, F, the I0 clinical scales, and the 15 new content scales. Profiles across administration modalities show a high degree of similarity, providing evidence for the comparability of computerized adaptive and conventional testing with the MMPI-2. Substantial item savings were found with the adaptive version. Future directions in the study of adaptive testing with the MMPI-2 are discussed. PMID- 16370885 TI - Factor analysis of MCMI-II and PACL basic personality scales in a college sample. AB - This study examined factor dimensions common to the eight basic personality scales of the Millon Clinical Multiaxial Inventory-II (MCMI-II; Millon, 1987) and Personality Adjective Check List (PACL; Strack, 1987, 1990). Subjects were 140 college students (65 men and 75 women). MCMI-II weighted raw scores (WRS) and WRS corrected for number of items endorsed true by regression were employed for analysis along with PACL T-scores. Principal components analyses with varimax and direct oblimin rotations were carried out separately on the two sets of MCMI-II and PACL scores. MCMI-II and PACL scales measuring the same personalities were usually correlated most highly with each other, although some divergences were noted. WRS yielded three bipolar dimensions and a fourth unipolar method factor that loaded only the five MCMI-II scales that were strongly correlated with number of items endorsed true. Residual scores yielded a more meaningful set of three bipolar dimensions labeled Social Introversion-Extraversion, Emotionality Restraint, and Social Dominance-Submissiveness, without the method factor, that were very similar to personality dimensions found separately in the two tests. More research is needed to clarify the response bias issue in the MCMI-II and to further explicate similarities and differences between the MCMI-II and the PACL. PMID- 16370886 TI - The Word Association Implications Test. AB - Three experiments (N = 144 college students) tested and documented the relationship between incidental learning of diagnostic schemata with performance on the Word Association Implications Test (WAIT). Moreover, Experiment 1 showed that instruction in diagnostic cues did not improve WAIT scores, although an incentive did. Experiment 2 supported the "developing schemata" concept by showing that subjects need to interpret WAIT protocols before they can generate possible diagnostic clues. Experiment 3 showed that inductive reasoning and viewing the task as more play- and game-like (vs. work- and test-like) predicted WAIT scores over and above the influence of incidental learning of schemata, in sum, results suggest that an heuristic discovery-oriented approach is most successful in learning "to read between the lines" of word associations. Discussion addresses the value of heuristic versus algorithmic approaches to the learning of personality assessment. PMID- 16370888 TI - Defense mechanisms in minority African-American and Hispanic youths: standardization and scale reliabilities. AB - The Defense Mechanisms Inventory (DMI), which has been standardized for White populations and has been used frequently in a variety of personality and interpersonal interaction studies, was administered to 229 minority noncollege youths. Reliability and sex difference data are presented, and the pattern of defenses is compared to published patterns among the White population. Normative tables are presented as references for future DMI studies with minorities, with comparisons for archival data on 504 college students. Data analyses demonstrate the reliability of the DMI for minorities, with tendencies toward the internalizing polarity of the composite (object relations) measure. Evidence is also presented that the DMI is more salient for the minority population on the composite cluster, rather than for the specific defenses. PMID- 16370889 TI - Test of structural relationships within a proposed exercise and self-esteem model. AB - This study tested structural relationships within a model proposed to explain the manner in which self-esteem changes are associated with exercise experiences. As initially assessed by self-efficacies (EFFs) specific to physical tasks within a training program, we postulated that competence can generalize to feelings of global self-esteem (SE) through an intervening construct of perceived physical competence (PC). Three measures of EFF and two each of PC and SE were administered to 145 people in their mid- to late adulthood. Confirmatory factor analysis examined orthogonal and oblique versions of three measurement models and identified three distinct but correlated factors, each assessed by its hypothesized indicators. Alternative structural equation models were specified and tested using both normal and nonnormal estimation procedures. The proposed model provided the most parsimonious fit and explained 29% of the variance in SE. Confirmation of its structural relationships provides preliminary validity for model use in examining the manner in which exercise experiences influence levels of self-perception. PMID- 16370890 TI - A short form of the Expanded Attributional Style Questionnaire. AB - A short (12 event) form of the Expanded Attributional Style Questionnaire was developed. In two studies, the subscales of the short form had correlations of .88 to .94 with those of the long form, similar reliabilities, and similar correlations with measures of depression. PMID- 16370891 TI - Developments in research in personality assessment. AB - Progress in personality research requires an adequate data analysis link between concepts of what people are like and observations of how they are likely to behave. Advances in computer technology have strengthened this link but also have made it necessary for investigators to be wary of lack of selectivity and trivialization in their research designs. PMID- 16370892 TI - Response to Exner's comments on "The Rorschach M response: a return to its roots". PMID- 16370893 TI - Parental authority questionnaire. AB - A questionnaire was developed for the purpose of measuring Baumrind's (1971) permissive, authoritarian, and authoritative parental authority prototypes. It consists of 30 items per parent and yields permissive, authoritarian, and authoritative scores for both the mother and the father; each of these scores is derived from the phenomenological appraisals of the parents' authority by their son or daughter. The results of several studies have supported the Parental Authority Questionnaire as a psychometrically sound and valid measure of Baumrind's parental authority prototypes, and they have suggested that this questionnaire has considerable potential as a valuable tool in the investigation of correlates of parental permissiveness, authoritarianism, and authoritativeness. PMID- 16370898 TI - Predicting the performance of government security personnel with the IPI and MMPI. AB - The Inwald Personality Inventory (IPI) and Minnesota Multiphasic Personality Inventory (MMPI) were administered to 307 newly hired government security personnel. After 9 to 12 months on the job, each was rated by their immediate supervisor on a 4-point scale of global performance (exceptional, satisfactory, unsatisfactory, or very unsatisfactory). Discriminant function analyses indicated that the IPI was superior to the MMPI in predicting later job performance, but the greatest prediction accuracy was found in discriminant functions based upon both tests together. However, the costs of falsely identifying candidates as poor risks and screening out those who would have been successful increased along with accuracy in predicting the true risks. It was concluded that organizations must ultimately decide upon the degree to which they are willing to sacrifice candidates who may have been successful in order to screen out those who are unsuitable. The implications of psychological screening for organizational selection policy are discussed. PMID- 16370900 TI - Changes in self- and object representation as revealed by reflection response. AB - This article considers the three Rorschach tests obtained over an 8-year period for one female patient who struggled around her image of herself and others. Her struggle is revealed in the vicissitudes of the reflection responses over the three testings. A formulation is advanced regarding the meaning and significance of the reflection response as an indication of an emerging self-experience. PMID- 16370901 TI - An attachment classification of desperate love. AB - The assessment utility of attachment theory constructs for research and diagnostic purposes is being increasingly explored and demonstrated. This study focuses on the association between desperate love and a modification of Ainsworth, Blehar, Waters, and Wall's (1978) taxonomy of attachment styles, hypothesizing that the experience of desperate love will be positively associated with the dependent and resistant/ambivalent attachment styles with no gender differences. The findings based on multivariate regression analyses generally corroborate the attachment style hypotheses, although with significant gender differences. Discussion focuses on the possible contrasts in the intrapsychic experience of desperate love for men and women as well as indications of causal trends for future research. PMID- 16370902 TI - Clinical assessment of object relations using the TAT. AB - Despite its widespread use, the Thematic Apperception Test (TAT) has rarely been used systematically as a clinical instrument. This article describes a multidimensional measure devised for the clinical assessment of object relations using the TAT. Four scales assessing different dimensions of object relations, which have been validated in several studies, are described: Complexity of Representations of People, Affect-Tone of Relationship Paradigms, Capacity for Emotional Investment in Relationships and Moral Standards, and Understanding of Social Causality. Use of the scales is illustrated by applying them to TAT responses of a borderline patient and a normal subject. PMID- 16370903 TI - Affect in early memories of borderline patients. AB - Psychoanalytically oriented observers using the framework of object relations theory noted that patients with borderline personality organization manifest serious pathology in their capacity to use internalized imagery to cope with the vicissitudes of separation-individuation. This study examined one source of such imagery, the affect associated with a specific sample of early memories, including the earliest recollections of parents. Patients with borderline personality disorder were compared with paranoid schizophrenics and patients with neurotic character pathology. Forty-five outpatients, 15 in each group, participated in the study. It was found that borderline patients had significantly fewer positively toned early memories than neurotics or paranoid schizophrenics. Only the neurotic group, had a majority of affectively positive memories. The results are generally consistent with object relations theories of borderline psychopathology. PMID- 16370904 TI - An examination of the construct validity of the Social Anhedonia Scale. AB - Two studies bearing on the construct validity of the Social Anhedonia Scale (SA) are reported. Study 1 reported convergent validity coefficients between the SA scale and numerous measures of traits reflecting a generalized attitude toward others (e.g., social interest and empathy) as well as several traits reflecting maladjustment without a social component as a test of discriminant validity (e.g., pessimism and rigidity). Study 1 found impressive convergent validity for the SA scale and mixed discriminant validity. Study 2 found that the SA scale was related to two dimensions of striving for security, one of which is strongly influenced by negative interpersonal tendencies, suggesting support for the validity of the SA scale. Implications for the measurement and concept of social anhedonia are presented. PMID- 16370905 TI - The relationship between parental divorce and the child's body boundary definiteness. AB - The relationship between parental divorce and body boundary definiteness was studied in a group of Swedish children. Rorschach tests were administered to a nonclinical sample of 90 school children between the ages of 10 and 12. Forty-six of the children's parents were divorced (hereafter called divorce children), and 44 of the children's parents were not divorced (hereafter called nondivorce children). Divorce children were interviewed after the testing. Nondivorce children were found to have firmer body boundaries (i.e., higher barrier and lower penetration scores) than divorce children. The quality of the child's relationship with the noncustodial father and the child's age at the time of separation were found to be related to the barrier score among boys but not among girls. The vulnerability of body boundaries appears to persist even years after the family breakup, indicating that for children of either sex the effects of divorce are long-term. PMID- 16370906 TI - Where's the beef? Putting personality back into personality assessment. AB - This article attempts to demonstrate the value of using a psychoanalytic theory of personality for psychological testing. This approach has more clinical utility than a solely research-based one. It recasts test data into conceptually related constructs that have internal consistency to each other and are directly relevant to psychotherapeutic treatment. Such theoretical recasting serves an organizing function, an integrative function, a clarification function, and a predictive function for the clinical inference process. Furthermore, a psychoanalytically oriented approach to testing allows for the expansion in sources of data that one considers in the testing situation. Five different sources of data emerge from the testing situation once one refocuses on theoretical constructs rather than test signs. These include test scores, test content, the patient-examiner interaction, patient behavior, and examiner countertransference. PMID- 16370907 TI - The role of elevation, scatter, and shape in MMPI profiles. AB - The contribution of profile elevation, scatter, and shape to differences between Minnesota Multiphasic Personality Inventory (MMPI) profiles was studied in psychiatric, normal, and mixed samples. Average between profile differences were smallest within the normal sample and largest within the mixed sample. Overall, the role of shape was most important, with elevation making a smaller but significant contribution to profile differences. Scatter was of minimal importance. Systems proposed by Skinner (1976) and Lorr (1982) were found to account for a large part of the variance due to shape, and these systems also were successful in classifying individual MMPI profiles. Advantages and applications of such systems in the analysis and description of profile data were discussed. PMID- 16370908 TI - On shortening the Expanded Attributional Style Questionnaire. AB - Whitley (in press) described a short form of the Expanded Attributional Style Questionnaire (EASQ). The EASQ was created to remedy the unsatisfactory psychometric properties of the Attributional Style Questionnaire; therefore, the benefits of a short form must be balanced with the possible costs, lower reliability and, hence, lower validity. Data from eight different samples using the EASQ (N = 963) suggest that the best items on the EASQ differ from sample to sample and from dimension to dimension. Further, reliabilities are reduced according to the predictions of the Spearman-Brown formula. Implications of these findings for attributional style research are discussed. PMID- 16370909 TI - Protein therapeutics--lessons learned and a view of the future. AB - Protein therapeutics represent a rapidly growing proportion of marketed drugs and have an undisputed place alongside chemistry-based oral therapies; indeed, for certain indications they are the only effective therapy. Therapeutic proteins can be mined from diverse sources to target interactions that are not accessible to small molecules, and can be engineered to have optimal pharmacological properties. Nevertheless, the development of such therapeutics is hampered by several issues, such as cost of production, patient compliance, immunogenicity and reticence of reimbursement agencies to pay for their use in chronic treatment. Herein we review some of these issues in detail. The application of technological advancements will address some of these issues and enable the development of a growing number of biological therapies that will continue to improve patient quality of life for decades to come. PMID- 16370910 TI - Regeneration of teeth using stem cell-based tissue engineering. AB - Tooth autotransplantation, allotransplantation and dental implants have existed for many years, but have never been totally satisfactory. Thus, the development of new methods of tooth replacement has become desirable, and with the increasing knowledge of stem cell biology becomes a realistic possibility. Stem cell-based tissue engineering involving the recapitulation of the embryonic environment demonstrates that dental, non-dental, embryonic and adult stem cells can contribute to teeth formation in the appropriate setting. Evidence that stem cell populations may be present in human teeth provides the opportunity to consider biological tooth replacement 'new for old'. PMID- 16370911 TI - Stem cells in inflammatory demyelinating disorders: a dual role for immunosuppression and neuroprotection. AB - In recent years much excitement has been generated over the possibility that adult stem cells may attempt repair of the injured central nervous system (CNS), thus setting the stage for their utilisation in the treatment of neurodegenerative disorders. Recent studies have shown that some subsets of stem cells can also modulate the (auto)immune response, thus providing a rationale for their use as therapy for experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis (MS). This article reviews the scientific evidence supporting the possible use of neural stem cells (NSCs) and mesenchymal stem cells (MSCs) for the treatment of MS. In addition, possible mechanisms sustaining the beneficial mode of action of haematopoietic stem cells (HSCs) following transplantation in MS individuals are discussed. Overall, it is proposed that limited subsets of adult stem cells may have a dual function that may be effective for the treatment of MS, an autoimmune disease of the CNS where degeneration of neural cells follows inflammation. PMID- 16370912 TI - Pancreatic islet transplantation for treating diabetes. AB - Pancreatic islet transplantation is one of the options for treating diabetes and has been shown to improve the quality of life of severe diabetic patients. Since the Edmonton protocol was announced, islet transplantation have advanced considerably, including islet after kidney transplantation, utilisation of non heart-beating donors, single-donor islet transplantation and living-donor islet transplantation. These advances were based on revised immunosuppression protocols, improved pancreas procurement and islet isolation methods, and enhanced islet engraftment. Further improvements are necessary to make islet transplantation a routine clinical treatment. To synergise efforts towards a cure for type 1 diabetes, a Diabetes Research Institute (DRI) Federation is currently being established to include leading diabetes research centres worldwide, including DRIs in Miami, Edmonton and Kyoto among others. PMID- 16370913 TI - Selective delivery of therapeutic agents for the diagnosis and treatment of cancer. AB - Research activity aimed towards achieving specific and targeted delivery of cancer therapeutics has expanded tremendously in the last decade, resulting in new ways of directing drugs to tumours, as well as new types of drugs. The available strategies exploit differences in the nature of normal and cancer cells and their microenvironment. The discovery and validation of cancer-associated markers, as well as corresponding ligands, is pivotal for developing selective delivery technology for cancer. Although most current clinical trials are either monoclonal antibody- or gene-based, methodological advances in combinatorial libraries of peptides, single chain variable fragments and small organic molecules are expected to change this scenario in the near future. Nanotechnology platforms today allow systematic and modular combinations of therapeutic agents and tumour-binding moieties that may generate novel, personalised agents for selective delivery in cancer. This paper discusses recent developments and future prospects of targeted delivery technologies in the management of cancer. PMID- 16370914 TI - Adenovirus p53 gene therapy. AB - To date, dysfunctional tumour suppressor genes are the most common genetic lesions identified in human cancers. Functional copies of tumour suppressor genes can be introduced into cancer cells by gene transfer using adenoviral vectors. This approach has been extensively studied in the clinic with intratumoural injection of a replication-defective adenovirus that expresses p53 (Ad-p53). Overexpression of p53 in cancer cells induces growth arrest and apoptosis. Ad-p53 injections have an excellent safety profile, and have mediated tumour regression and growth arrest as monotherapy, or have overcome resistance or increased the effectiveness of radiation therapy and chemotherapy. Expression of the p53 transgene has occurred at high levels and is associated with the activation of other genes in the p53 pathway. These studies indicate proof-of-principle for tumour suppressor gene therapy and represent a new paradigm in targeted therapy. PMID- 16370915 TI - Therapeutic potential of adenovirus as a vaccine vector for chronic virus infections. AB - Therapeutic vaccines for chronic infections and cancer are needed. Challenges faced by therapeutic vaccines differ from those of preventative vaccines. Whereas the latter target a naive immune system, the former have to readjust an antigen experienced immune system that is subverted due to sustained exposure to antigen. E1-deleted adenoviral vectors have succeeded preclinically as preventative vaccines and are now in clinical trials. Their potential as therapeutic vaccines for diseases caused by chronic virus infections or virus-associated malignancies remains to be explored in more depth and may require modifications to circumvent negative immunoregulatory pathways that develop following chronic infections or during tumour progression. PMID- 16370916 TI - The emerging role of recombinant activated Factor VII (rFVIIa) in the treatment of blunt traumatic haemorrhage. AB - Recombinant activated Factor VII (rFVIIa; eptacog alpha [activated], NovoSeven) is currently used for the management of a subgroup of haemophilia patients with inhibitors to Factors VIII or IX, and is under investigation as an adjuvant therapy for critical bleeding from other causes, including trauma. rFVIIa has a mode of action founded on physiological coagulation processes, and causes localised haemostasis at injury sites, both spontaneous and traumatic, with the capacity to correct the systemic coagulopathy associated with massive blood loss and its management. This review charts the development of rFVIIa as a new and potent adjuvant therapy for severe bleeding and coagulopathy caused by blunt trauma, where it is reported to produce rapid and significant haemostasis, reducing transfusion requirements and improving clinical outcome. PMID- 16370917 TI - Pharmacological strategies for the prevention of Alzheimer's disease. AB - This review examines key pharmacological strategies that have been clinically studied for the primary or secondary prevention of Alzheimer's disease. Much information (neuropsychological, genetic and imaging) is already available to characterise an individual's risk for developing Alzheimer's disease. However, regulatory pathways for obtaining a prevention indication are less well charted, and such trials tend to involve 3- to 7-year studies of 1000 - 5000 individuals, depending on baseline status. Treatments developed for prevention will also need to have superior safety. For these reasons, > 100 proprietary pharmacological products are currently being developed for an Alzheimer's disease treatment, but only a few are being studied for prevention. Randomised trial data are available for antihypertensive agents (calcium channel blockers, angiotensin-converting enzyme inhibitors), pravastatin, simvastatin, conjugated oestrogen, raloxifene, rofecoxib, CX516 (AMPA agonist) and cholinesterase inhibitors regarding efficacy for Alzheimer's disease prevention. At least four large prevention trials of conjugated oestrogen, selenium and vitamin E, Ginkgo biloba and statins are currently underway. Strategies using other agents have not yet been evaluated in Alzheimer's disease prevention clinical trials. These include anti-amyloid antibodies, active immunisation, selective secretase inhibitors and modulators, microtubule stabilisers (e.g., paclitaxel), R-flurbiprofen, xaliproden, ONO-2506, FK962 (somatostatin releaser), SGS 742 (GABA(B) antagonist), TCH 346 (apoptosis inhibitor), Alzhemedtrade mark, phophodiesterase inhibitors, rosiglitazone, leuprolide, interferons, metal-protein attenuating compounds (e.g., PBT2), CX717, rasagaline, huperzine A, antioxidants and memantine. Studies combining lifestyle modification and drug therapy have not been conducted. Full validation of surrogate markers for disease progression (such as amyloid imaging) should further facilitate drug development. Reducing the complexity of prevention trials and gaining regulatory consensus of design is a high priority for the field. PMID- 16370918 TI - Pharmacological management of sleep apnoea. AB - Obstructive sleep apnoea poses a significant health hazard that is associated with leading causes of mortality and morbidity. Nasal continuous positive airway pressure is the primary treatment modality, with surgical treatments as alternatives. Oral appliances and pharmacological therapy remain adjunctive modalities. Non-specific treatments include weight loss, postural therapy and behavioural measures. Pharmacotherapy goals include the reduction of risk factors for sleep apnoea; correction of underlying predisposing metabolic diseases, such as hypothyroidism or acromegaly; treatment of associated symptoms, including excessive daytime sleepiness; and prevention of apnoeas/hypopnoeas. This paper reviews data supporting the treatment of sleep apnoea with various pharmacological agents, including intranasal corticosteroids, decongestant sprays, nicotine therapy, opiate antagonists, methylxanthine derivatives, oestrogen and progesterone, testosterone, thyroid hormone, growth hormone therapy for acromegaly, beta-blockers, alpha-adrenergic agonists, angiotensin-converting enzyme inhibitors, glutamate antagonists, acetazolamide, selective serotonin re uptake inhibitors, tricyclic antidepressants, physostigmine, modafinil and TNF alpha antagonists, in addition to supplemental oxygen, and carbon dioxide inhalation. Some of these drugs have received very little testing and are the subject of few research articles. PMID- 16370919 TI - Management of bleeding emergencies: when to use recombinant activated factor VII. AB - Recombinant activated Factor VII (rVIIa) was originally released as a clotting factor, with use limited to a select group of patients who had few other treatment options. Due to the apparent ability of rVIIa to stop bleeding, no matter what the underlying cause, there is great interest in use of rVIIa in a wide range of bleeding patients. This article discusses rVIIa and its uses in a variety of patients, especially liver disease and trauma patients, and makes suggestions for appropriate use. Although most of the reports on rVIIa are anecdotes and case series, there is increasing data for clinical trials to help guide usage. PMID- 16370920 TI - Pharmacological management of transient synovitis. AB - Synovitis is a painful and, occasionally, disabling disease. Patients with synovitis, especially new onset synovitis, should be referred to a rheumatologist urgently so that they can be assed and treated as quickly as possible. Clinical assessment and investigations are required to help differentiate between transient (< 3 months) and persistent (> 3 months) synovitis. This differentiation is important, as persistent synovitis can lead to joint damage and disability. Septic arthritis is a rheumatological emergency requiring immediate assessment and specific treatment. The earlier synovitis is treated, the more effective treatment is likely to be. If treated very early, there is potential to prevent the move from transient to persistent synovitis. Transient synovitis can be treated with painkillers, NSAIDs and/or corticosteroids, depending on severity. Persistent synovitis may also require disease-modifying drugs. Clinical indicators of persistence include symptom duration at first visit, early morning stiffness for > 1 h, arthritis in more than three joints, bilateral compression pain in metatarsophalangeal joints, rheumatoid factor positivity, anti-cyclic citrullinated peptide antibody positivity, erosions on hand or feet X-rays and a family history of rheumatoid arthritis. Disease modifying drugs need to be considered early to achieve clinical remission before damage and disability occur. Despite emerging new treatments for synovitis, especially persistent synovitis, full clinical remission is still not achieved in most patients, and more research into disease processes and targeted therapies is required. PMID- 16370921 TI - An update on the available treatments for non-erosive reflux disease. AB - Non-erosive reflux disease is defined as the presence of troublesome reflux symptoms, such as heartburn and regurgitation, in the absence of endoscopically visible damage of the oesophageal mucosa. In comparison with erosive oesophagitis, non-erosive reflux disease is the most common clinical manifestation of gastro-oesophageal reflux disease. Pathophysiologically, it is not a homogeneous disease as only approximately two-thirds of patients have truly acid-related symptoms. This explains the fact that patients with non-erosive reflux disease consistently show a poorer response to proton pump inhibitor treatment than patients with erosive oesophagitis. Nevertheless, profound acid inhibition by proton pump inhibitors is the recommended first-line treatment in patients suffering from this condition, both in the initial phase and for long term care. Non-responders to proton pump inhibitor therapy should be subjected to a thorough examination and treated on an individual basis. Emerging data on the long-term course of reflux disease under routine clinical care have led to the adoption of new therapeutic strategies that would have been unthinkable only a few years ago. PMID- 16370922 TI - Do health-status measures play a role in improving treatment in chronic obstructive pulmonary disease? AB - During the past few decades, health status has become increasingly important in the clinical research of chronic obstructive pulmonary disease. The use of health status questionnaires in routine practice can enhance understanding about the impact of the disease on the patient, improve standardisation and increase compliance through increased patient satisfaction. However, before health-status measurement in individual patients can be used in routine practice, questionnaires have to be validated on an individual level. In this article, the authors suggest a new method of assessing this individual validity, to enhance the use of health-status instruments in daily clinical practice, and thus improve treatment in chronic obstructive pulmonary disease. PMID- 16370923 TI - Perindopril. AB - Perindopril is a third-generation ACE inhibitor that is characterised as a small, lipophilic molecule with a therapeutically active carboxyl side group. These and other features combine to make this a unique member of a very well-established class of drugs that have proven efficacy in a wide range of cardiovascular diseases. The Perindopril Protection Against Recurrent Stroke Study (PROGRESS) demonstrated benefit in the secondary prevention of patients with stroke, whereas the Perindopril and Remodelling in Elderly with Acute Myocardial Infarction (PREAMI) trial supports extended routine use after myocardial infarction. The most recent evidence from the European Trial on Reduction of Cardiac Events with Perindopril in Stable Coronary Artery Disease (EUROPA) and the Anglo-Scandinavian Cardiac Outcomes Trial (ASCOT) show that perindopril is able to improve the prognosis of patients with a relatively low global cardiovascular risk, denoted either by the presence of stable coronary artery disease or of essential hypertension in conjunction with at least three other risk factors. The fact that major relative risk reductions have been reported for these two studies is matched by the significance of the findings to modern clinical practice. Both studies were conducted in the context of advance concomitant care that is typically better in clinical trials than in routine practice. In particular, the benefits observed were seen to be of a similar magnitude, and also independent of those resulting from statin therapy. Of particular interest is the likely complimentary action of these treatment strategies with regard to the stabilisation of atheromatous plaques. Perindopril is a well-established drug, the full value of which is only now becoming fully apparent. PMID- 16370924 TI - Tirofiban for the treatment of ischaemic stroke. AB - Tirofiban is one of three glycoprotein IIb/IIIa receptor antagonists approved by the US FDA, beside abciximab and eptifibatide. The approval of tirofiban covers conservative treatment of myocardial infarction and unstable angina, as well as percutaneous coronary intervention, for which treatment with tirofiban is recommended in moderate-to-high-risk patients. The efficacy of glycoprotein IIb/IIIa antagonists in myocardial infarction indicated that these agents may also be helpful in the treatment of acute ischaemic stroke. Although experimental data are lacking, observational studies are promising. In recent years, increasing effort in studying glycoprotein IIb/IIIa antagonists has been made, mostly for treatment with abciximab. However, there is one Phase II trial that investigated treatment with tirofiban. PMID- 16370925 TI - Argatroban, a direct thrombin inhibitor for heparin-induced thrombocytopaenia: present and future perspectives. AB - Heparin remains the most commonly used anticoagulant in the treatment of patients with acute vascular syndromes, including myocardial infarction, unstable angina and ischaemic stroke. However, heparin therapy is not always associated with a significant improvement of clinical outcomes, is linked with enhanced bleeding risk and can occasionally provoke the development of heparin-induced thrombocytopaenia, the most devastating complication of conventional therapy with unfractioned heparin. Understanding the key role of thrombin in clot formation and platelet activation has stimulated the development of a new class of drugs - direct thrombin inhibitors. The direct thrombin inhibitor argatroban has been known for decades. Similar to the unfractioned heparin, argatroban requires intravenous administration and activated partial prothrombin time-dependent dose adjustment; however, this pharmacological agent has a relatively short half-life that broadens its safety margins, as well as its low antigenic potential due to the small molecular weight of the compound. The efficacy of argatroban has been demonstrated among patients with acute coronary syndromes and stroke. However, this drug is currently approved by the FDA only for the treatment of patients with heparin-induced thrombocytopaenia. Indeed, in such patients, argatroban significantly improves clinical outcomes, and is associated with reduced mortality. Further clinical studies are needed to present more clinical evidence necessary to broad the indication spectrum of this agent. PMID- 16370926 TI - Tacrolimus in rheumatoid arthritis. AB - Tacrolimus is an immunosuppressive drug that has been used widely in organ transplantation and topically for atopic dermatitis. Tacrolimus exerts its immunosuppressive effects by the inhibition of calcineurin, leading to interference with T-cell activation. As T-cell activation plays a major role in the pathogenesis of rheumatoid arthritis, there has been an interest in the use of tacrolimus for the treatment of rheumatoid arthritis. The pharmacological properties of tacrolimus have the potential of suppressing the production of inflammatory cytokines, improvement of joint inflammation, improvement of bone and cartilage destruction, improvement of functional status and relief from arthritic pain. This article reviews the pharmacodynamics, pharmacokinetics, clinical efficacy, safety and role of tacrolimus in the treatment of rheumatoid arthritis. PMID- 16370927 TI - An update on the use of ophthalmic ketorolac tromethamine 0.4%. AB - Ketorolac tromethamine 0.4% ophthalmic solution, a recent reformulation of the original ketorolac tromethamine 0.5% solution, is indicated for the reduction of ocular pain and burning/stinging following cataract and refractive surgery. Studies have demonstrated that ketorolac tromethamine 0.4% has equivalent efficacy to ketorolac tromethamine 0.5% in reducing postsurgical inflammation and controlling pain. Several studies have demonstrated that, as well as reducing pain and ocular inflammation, ketorolac tromethamine 0.4% effectively treats cystoid macular oedema, inhibits miosis and may prevent cystoid macular oedema when used both pre- and postoperatively. Ketorolac tromethamine 0.4% is a versatile agent and is effective when used as either monotherapy or as an adjunct therapy to steroids. PMID- 16370928 TI - Is exenatide advancing the treatment of type 2 diabetes? AB - Glucagon-like peptide 1 is an intestinal peptide hormone that is secreted in response to food to regulate the postprandial blood glucose concentration. Exendin-4 is a 39-amino acid peptide that acts as an agonist at the glucagon-like peptide 1 receptor. Synthetic exendin-4 (exenatide) has recently been trialled in patients with Type 2 diabetes taking either metformin alone or a combination of metformin and a sulfonylurea. In both trials, exenatide 5 and 10 microg s.c. was shown to improve glycaemic control, with few adverse events. Exenatide represents a new and useful addition to the medicines used to treat Type 2 diabetes. PMID- 16370929 TI - Novel cholinesterase inhibitors as future effective drugs for the treatment of Alzheimer's disease. AB - Current pharmacotherapy for Alzheimer's disease involves compounds that are aimed at increasing the levels of acetylcholine in the brain by facilitating cholinergic neurotransmission through inhibition of cholinesterase. These drugs, known as acetylcholinesterase inhibitors, have been shown to improve cognition and global functions but have little impact on improving the eventual progression of the disease; however, there is evidence that other cholinesterases such as butyrylcholinesterase can play an important role in cholinergic function in the brain, and the long-suspected non-cholinergic actions of acetylcholinesterase, mainly the interference with the beta-amyloid protein cascade, have recently driven a profound revolution in cholinesterase drug research. Several disease modifying agents are under development that target these enzymes and have hope of becoming the next generation of effective drugs in the treatment of Alzheimer's disease. PMID- 16370930 TI - Oral amphipathic peptides as therapeutic agents. AB - Cholesterol can promote inflammation by its ability to stimulate the production of reactive oxygen species that result in the formation of pro-inflammatory oxidised phospholipids. High-density lipoproteins (HDLs) are part of the innate immune response and can be either pro- or anti-inflammatory independently of plasma HDL-cholesterol levels. During systemic inflammation as occurs with atherosclerosis, Apolipoprotein A-I can be altered, reducing its ability to promote reverse cholesterol transport and HDL can become pro-inflammatory. Amphipathic peptides with either a class A amphipathic helix (D-4F) or a class G* amphipathic helix (D-[113-122]apoJ), or even those that are too small to form a helix (KRES and FREL) have some similar characteristics. Their interaction with lipids leads to a reduction in lipoprotein-lipid hydroperoxides that releases HDL associated antioxidant enzymes, such as paraoxonase, therefore providing antiatherosclerosis and anti-inflammatory activity. In addition, the peptide D-4F stimulates the formation and cycling of pre-beta HDL. These amphipathic peptides appear to have therapeutic potential as oral agents. PMID- 16370931 TI - Dual action of the inhibitors of cyclin-dependent kinases: targeting of the cell cycle progression and activation of wild-type p53 protein. AB - The inhibition of cyclin-dependent kinases (CDKs) represents a novel approach to the therapy of human malignancies. Already in clinical trials, recently developed CDK inhibitors very efficiently target the rapidly proliferating cancer cells and inhibit their cell-cycle progression. Interestingly, some CDK inhibitors additionally affect the stability and activity of the tumour-suppressor protein p53, thereby enhancing their antiproliferative action towards cancer cells. Considering the fact that the p53 protein is mutated or inactivated in approximately 50% of all human cancers, the efficacy of CDK inhibitor therapy could differ between cancer cells depending on their p53 status. Moreover, recent reports demonstrating that some cancer cells can proliferate despite CDK2 inhibition questioned the central role of CDK2 in the cell-cycle control and suitability of CDK2 as a therapeutic target; however, the p53 activation that is mediated by CDK inhibitors could be essential for the efficacy of CDK inhibitors in therapy of CDK2-independent cancers. Furthermore, there is also reason to believe that CDK2 inhibitors could be used for another purpose, to protect normal cells from the effects of chemotherapy. PMID- 16370932 TI - Pharmacological management of undifferentiated spondyloarthropathies. AB - Among undifferentiated spondyloarthropathies (uSpA) are included incomplete forms or early phases of definite seronegative spondyloarthritides and cases of spondyloarthritides that remain undifferentiated. The treatment of all spondyloarthritides is more conditioned by the disease localisation, number of involved districts and severity of inflammation rather than by the subtype of the spondyloarthritides itself. Specific studies focused on the pharmacological approach to uSpA are scarce. As a consequence, many drugs are used on the basis of the experiences that have been gained in the treatment of other spondyloarthritides. At present, non-steroidal anti-inflammatory drugs/COX-2 inhibitors and disease-modifying antirheumatic drugs are the main therapeutic agents for the involvement of peripheral joints in uSpA. In those cases in which severe symptoms persist despite these treatments or when there is a severe axial involvement, antitumour necrosis factor agents represent an effective choice. In these patients, antitumour necrosis factor treatment raises the important possibility of blocking a shift from uSpA to differentiated severe forms of spondyloarthritides such as ankylosing spondylitis. Local administration of corticosteroids is useful when a small number of joints are involved, as is also the case for the extra-articular localisations of soft tissues. PMID- 16370933 TI - Exogenous surfactant therapy for ARDS. AB - Regardless of the cause, a common pathophysiological feature of patients with acute respiratory distress syndrome is a dysfunction of the endogenous surfactant system. Although exogenous surfactant therapy has proven to be an effective treatment for neonatal respiratory distress syndrome, no similar current effective therapy exists for patients with acute respiratory distress syndrome. This is mainly due to the complexity of the lung injury that is involved with this disorder. Results from clinical trials, to date, have failed to show an improvement in patient survival after administration of exogenous surfactant; however, ongoing and future research efforts suggest that this therapy may eventually be feasible. PMID- 16370934 TI - Tipranavir: a protease inhibitor for multi-drug resistant HIV-1. AB - Since the mid-1990s, combination therapies to treat HIV-1 infection have greatly reduced morbidity and mortality from AIDS in developed countries with access to the medications. However, the development of viral resistance to available antiretrovirals is one of the many limitations to therapy that has emerged. Of the 24 licensed antiretroviral medications and medication combinations in the US, tipranavir is one of the few agents to specifically target highly treatment experienced patients with multi-drug resistant HIV-1. It displays activity against the virus that is cross-resistant to other protease inhibitors. In this review, issues in treating multi-drug resistant HIV-1 and the potential clinical utility of tipranavir in the US will be discussed. PMID- 16370935 TI - Rubitecan. AB - The only approved camptothecins for use in patients to date (topotecan and irinotecan) are delivered intravenously. Thus, an oral camptothecin analogue that would provide the convenience of oral delivery with the flexibility for a variety of prolonged treatment schedules would be advantageous. Rubitecan is an orally available camptothecin analogue that also has potential for delivery transdermally or by inhalation. Like all of the camptothecins, its antitumour activity is mediated through the inhibition of DNA topoisomerase I, which is involved in relaxing supercoiled DNA, which is important for the process of DNA replication and RNA transcription. Rubitecan exists in equilibrium as 9-nitro camptothecin (9-NC) and 9-amino-camptothecin (9-AC), a metabolite that is thought to be active although it failed in clinical trials. Both 9-NC and 9-AC contain a lactone ring that is required for optimal activity with the carboxylic acid (open ring) forms being significantly less active or inactive. A more acidic environment favours the lactone ring structure, whereas neutral or basic conditions favour the conversion to the carboxylic acid form. In addition to issues of lactone ring stability at physiological pH (true for all of the camptothecin analogues), there is pharmacokinetic variability that has had to be dealt with during the development of rubitecan. Preclinically, rubitecan has shown activity against a broad spectrum of tumour types in in vitro and in vivo human tumour xenograft models. Frustratingly, the level of activity of an agent in preclinical models has not always translated into similar activity against human tumours in clinical trials. To date, with the exception of pancreatic and possibly ovarian cancer, rubitecan has had disappointing activity against a number of other solid tumours in relatively small Phase I/II trials; however, it has shown sufficient activity against pancreatic cancer, a malignancy that remains difficult to treat, to continue to be evaluated in clinical trials for this indication. Results of clinical trials in the next few years should determine whether rubitecan can find a role in cancer therapy. PMID- 16370936 TI - Nuclear gene delivery: the Trojan horse approach. AB - The nuclear envelope represents a formidable barrier to the transfer of plasmids to the cell nucleus, particularly in nondividing cells. The probability of intact plasmids arriving in the nucleus by a passive process is extremely low. There is substantial evidence in the literature that describes the transport of macromolecules, including plasmids, to the nucleus as a very inefficient process, and so far attempts to affect the active transport through the nuclear pores have achieved limited success. Several approaches have been attempted to improve nuclear transport of plasmids, including the condensation of plasmids to unimolecular complexes of minimal hydrodynamic diameter to favour passive transport through the nuclear pore complex, and the incorporation of nuclear localisation signals in the plasmid or in the delivery system to enhance the active transport of plasmids through the nuclear pores. PMID- 16370937 TI - Bacterial ghosts as a novel advanced targeting system for drug and DNA delivery. AB - Although there are powerful drugs against infectious diseases and cancer on the market, delivery systems are needed to decrease serious toxic and noncurative side effects. In order to enhance compliance, several delivery systems such as polymeric micro- and nanoparticles, liposomal systems and erythrocyte ghosts have been developed. Bacterial ghosts representing novel advanced delivery and targeting vehicles suitable for the delivery of hydrophobic or water-soluble drugs, are the main focus of this review. They are useful nonliving carriers, as they can carry different active substances in more than one cellular location separately and simultaneously. Bacterial ghosts combine excellent natural or engineered adhesion properties with versatile carrier functions for drugs, proteins and DNA plasmids or DNA minicircles. The simplicity of both bacterial ghost production and packaging of drugs and/or DNA makes them particularly suitable for the use as a delivery system. Further advantages of bacterial ghost delivery vehicles include high bioavailability and a long shelf life without the need of cold-chain storage due to the possibility to freeze-dry the material. PMID- 16370938 TI - Single nucleotide polymorphisms in human P-glycoprotein: its impact on drug delivery and disposition. AB - Drug efflux pumps belong to a large family of ATP-binding cassette transporter proteins. These pumps bind their substrate and export it through the membrane using energy derived from ATP hydrolysis. P-glycoprotein, the main efflux pump in this family, is expressed not only in tumour cells but also in normal tissues with excretory function (liver, kidney and the intestine). It has a broad specificity of substrates and plays an important role in drug delivery and disposition. Recently, genetic screening of P-glycoprotein has yielded multiple single nucleotide polymorphisms, which seem to alter transporter function and expression. This review discusses the various polymorphisms of this gene and its impact on drug disposition and diseases. PMID- 16370939 TI - Gene delivery and gene therapy of prostate cancer. AB - Surgery, radiation or hormonal therapy are not adequate to control prostate cancer. Clearly, other novel treatment approaches, such as gene therapy, for advanced/recurrent disease are desperately needed to achieve long-term local control and particularly to develop effective systemic therapy for metastatic prostate cancer. In the last decade, significant progress in gene therapy for the treatment of localised prostate cancer has been demonstrated. A broad range of different gene therapy approaches, including cytolytic, immunological and corrective gene therapy, have been successfully applied for prostate cancer treatment in animal models, with translation into early clinical trials. In addition, a wide variety of viral and nonbiological gene delivery systems are available for basic and clinical research. Gene therapy approaches that have been developed for the treatment of prostate cancer are summarised. PMID- 16370940 TI - Tailoring antibodies for radionuclide delivery. AB - Therapeutic antibodies are well established as an important class of drugs in modern medicine. The exquisite specificity and affinity for a specific target offered by antibodies has also encouraged their development as delivery vehicles for agents such as radionuclides to target tissues, for radioimmunoimaging and radioimmunotherapy. Specifically, in nuclear medicine, radionuclide-conjugated antibody molecules make it possible to image diseased loci with greater sensitivity than other imaging modalities such as magnetic resonance imaging. Furthermore, two radionuclide-conjugated antibodies have recently been approved for the therapy of non-Hodgkin's lymphoma. However, optimal implementation of antibodies has been limited by the extended circulation persistence that is characteristic of native antibodies, which is responsible for increased background activity in radioimmunoimaging applications and dose-related normal organ toxicities in radioimmunotherapy. In this article the current status of radiolabelled intact antibodies is reviewed, focusing on strategies to improve their pharmacokinetic properties to suit a desired application. Examples from the literature that represent different approaches to accomplishing this task in terms of their successes as well as limitations, and perspectives for the future are discussed. PMID- 16370941 TI - Aerosolised antibiotics: a critical appraisal of their use. AB - Aerosolised antimicrobial agents have been used in clinical practice since the 1950s. The main advantage of this route of administration is the targeted drug delivery to the site of infection in the lung. Exploitation of this targeted delivery can yield high concentrations at the site of infection/colonisation while minimising systemic toxicities. It is important to note that the ability of a drug to reach the target area in the lung effectively is dependent on a number of variables, including the nebuliser, patient technique, host anatomy and disease-specific factors. The most convincing data to support the use of aerosolised antimicrobials has been generated with tobramycin solution for inhalation (TOBI, Chiron Corp.) for maintenance treatment in patients with cystic fibrosis. In addition to cystic fibrosis, the use of aerosolised antimicrobials has also been studied for the treatment or prevention of a number of additional disease states including non-cystic fibrosis bronchiectasis, ventilator associated pneumonia and prophylaxis against pulmonary fungal infections. Key studies evaluating the benefits and shortcomings of aerosolised antimicrobial agents in these areas are reviewed. Although the theory behind aerosolised administration of antibiotics seems to be sound, there are limited data available to support the routine use of this modality. Owing to the gaps still existing in our knowledge base regarding the routine use of aerosolised antibiotics, caution should be exercised when attempting to administer antimicrobials via this route in situations falling outside clearly established indications such as the treatment of patients with cystic fibrosis or Pneumocystis pneumonia. PMID- 16370942 TI - Noninvasive delivery technologies: respiratory delivery of vaccines. AB - This paper reviews the developments in noninvasive methods of drug delivery, with a focus on the delivery of vaccines via the respiratory tract. Recent results indicate that the respiratory system, and the nasal mucosa in particular, provide a valuable target site for immunisation against respiratory and mucosal pathogens. Vaccine delivery via the nasal and pulmonary routes each present distinct sets of performance requirements. Current delivery systems in development for both routes are reviewed herein. The storage and respiratory delivery of drugs and vaccines in powder form has been shown to provide improved stability and extended retention time in the respiratory mucosa. These features, in addition to the noninvasive nature of respiratory delivery, can provide benefits to public health vaccination campaigns, facilitating mass vaccination without the high cost of maintaining cold-chain storage. PMID- 16370943 TI - Development of supersaturatable self-emulsifying drug delivery system formulations for improving the oral absorption of poorly soluble drugs. AB - The supersaturatable self-emulsifying drug delivery system (S-SEDDS) represents a new thermodynamically stable formulation approach wherein it is designed to contain a reduced amount of a surfactant and a water-soluble cellulosic polymer (or other polymers) to prevent precipitation of the drug by generating and maintaining a supersaturated state in vivo. The S-SEDDS formulations can result in enhanced oral absorption as compared with the related self-emulsifying drug delivery systems (SEDDS) formulation and the reduced surfactant levels may minimise gastrointestinal surfactant side effects. PMID- 16370944 TI - Colon drug delivery. AB - Oral drug delivery to the colon has attracted significant attention during the past 20 years. Colon targeting is recognised to have several therapeutic advantages, such as the oral delivery of drugs that are destroyed by the stomach acid and/or metabolised by pancreatic enzymes. Sustained colonic release of drugs can be useful in the treatment of nocturnal asthma, angina and arthritis. Local treatment of colonic pathologies, such as ulcerative colitis, colorectal cancer and Crohn's disease, is more effective with the delivery of drugs to the affected area. Likewise, colonic delivery of vermicides and colonic diagnostic agents requires smaller doses. This article aims to provide an insight into the design and manufacturing considerations, and an evaluation of colonic drug delivery systems in order to understand why there are still few delivery technologies that have reached the market, despite intensive research in this field. For this purpose, various approaches to colon-specific drug delivery are discussed. PMID- 16370945 TI - Transdermal iontophoresis. AB - Iontophoresis is a technique used to enhance the transdermal delivery of compounds through the skin via the application of a small electric current. By the process of electromigration and electro-osmosis, iontophoresis increases the permeation of charged and neutral compounds, and offers the option for programmed drug delivery. Interest in this field of research has led to the successful delivery of both low (lidocaine) and high molecular drugs, such as peptides (e.g., luteinising hormone releasing hormone, nafarelin and insulin). Combinations of iontophoresis with chemical enhancers, electroporation and sonophoresis have been tested in order to further increase transdermal drug permeation and decrease possible side effects. In addition, rapid progress in the fields of microelectronics, nanotechnology and miniaturisation of devices is leading the way to more sophisticated iontophoretic devices, allowing improved designs with better control of drug delivery. Recent successful designing of the fentanyl E-TRANS iontophoretic system have provided encouraging results. This review will discuss basic concepts, principles and applications of this delivery technique. PMID- 16370946 TI - Polymeric micelles for drug delivery. AB - Polymeric micelles have been the subject of many studies in the field of drug delivery for the past two decades. The interest has specifically been focused on the potential application of polymeric micelles in three major areas in drug delivery: drug solubilisation, controlled drug release and drug targeting. In this context, polymeric micelles consisting of poly(ethylene oxide)-b poly(propylene oxide), poly(ethylene oxide)-b-poly(ester)s and poly(ethylene oxide)-b-poly(amino acid)s have shown a great promise and are in the front line of development for various applications. The purpose of this manuscript is to provide an update on the current status of polymeric micelles for each application and highlight important parameters that may lead to the development of successful polymeric micellar systems for individual delivery requirements. PMID- 16370947 TI - The 15th International Society of Aerosols in Medicine Congress. Perth, Australia, 14-18 March 2005. AB - This international meeting brought together approximately 250 delegates from the pharmaceutical industry, academia, hospitals and government agencies, to discuss the latest research and development on areas related to inhalation aerosols. Fundamental science and applied research encompassing both the biological and physicochemical aspects were presented. There was a wide range of topics covered, from immune modulation to pharmaceutical regulatory issues, including aerosol clearance; industry innovations; aerosols and in utero effects; technical advances in imaging; inhalation catastrophes; as well as recent advances and future directions in aerosol delivery systems. This biennial congress has provided an excellent forum for stimulating fruitful discussion of aerosols in medicine. PMID- 16370948 TI - COX-2 selective drugs and cardiovascular risks: same data but discrepant conclusions? PMID- 16370949 TI - Potential risk of probiotic usage. PMID- 16370950 TI - Over-the-counter herbals and drug interactions: getting the (right) message across. PMID- 16370951 TI - Herbal remedies and drug interactions: an issue overlooked by health authorities. PMID- 16370952 TI - Toxicity of infliximab in the course of treatment of Crohn's disease. AB - Infliximab is a monoclonal antibody directed against the pro-inflammatory mediator TNF-alpha, which was approved in the US in 1998 for treatment-resistant Crohn's disease. Since that time, the indications have dramatically expanded to include rheumatoid arthritis, ankylosing spondylitis, psoriasis and most recently, active ulcerative colitis. Although the safety profile in the initial studies was quite favourable, subsequent studies have shown that a small percentage of patients reported severe side effects, including pneumonia, tuberculosis, lymphoma, drug-induced lupus and hepatotoxicity. Although these complications are rare, it is important to properly screen patients for predisposing conditions before beginning treatment. Furthermore, concurrent use of other immunosuppresive agents, such as 6-mercaptopurine, may reduce the incidence of less serious side effects, such as arthralgias, myopathies and other antibody-associated diseases. Since its approval, infliximab has revolutionised the treatment of Crohn's disease and has shown benefit in a variety of other inflammatory conditions, but significant toxicities can occur that necessitate thorough screening protocols and periodic clinical evaluation. PMID- 16370953 TI - When science is not enough - a risk/benefit profile of thiomersal-containing vaccines. AB - Without a preservative, such as thiomersal (known as thimerosal in the US), multi dose liquid presentations of vaccine are vulnerable to bacteriological contamination that can result in death or serious illness of the recipient. Concerns about levels of mercury exposure from thiomersal-containing vaccines were first raised in the US during 1999 in the context of Hepatitis B vaccine for newborns. Since then, a large body of evidence from animal and epidemiological studies has accumulated on the safety of thiomersal. Ironically, these data have become largely irrelevant in wealthy countries, where mono-dose, thiomersal-free vaccines have been introduced as a precautionary measure in almost all childhood vaccines, in part related to residual public scepticism. In poor countries, multi dose vials remain important for vaccine delivery. There is a real danger that this controversy may result in the loss to the world of thiomersal as a preservative, simply from popular pressure. In reality, it would be impossible to cease overnight using thiomersal and maintain the supply of vital vaccines. This paper reviews and summarises the data available from published studies on mercury toxicity, and thiomersal in vaccines in particular, that overwhelmingly indicate continued use of thiomersal is safe in those countries where it is most needed. PMID- 16370954 TI - Allergic reactions to beta-lactams. AB - Allergy to beta-lactam antibiotics is the most frequent cause of drug-induced immunological reactions, although the prevalence is not exactly known. IgE- and T cell-dependent responses are the main mechanisms involved, although other immunological mechanisms can also participate, especially in haematological abnormalities, such as immune haemolytic anaemia or thrombocytopoenia. Aside from their frequency, the clinical entities reported nowadays have changed little since penicillin was first used. The variation in beta-lactams consumption through the year has modified the pattern and specificities of allergic reactions for IgE and T cell responses. Benzylpenicillin is no longer the beta-lactams most frequently prescribed and other chemical structures, with new or modified haptens, have progressively replaced it. This is relevant for the diagnostic evaluation and management of beta-lactam hypersensitivity. PMID- 16370955 TI - Medication overuse headache. AB - Medication overuse headache (MOH) has developed into the third most common type of headache after tension-type headache and migraine. The prevalence of MOH is approximately 1% of the world's population and it shows an increasing trend as recent studies reveal a common involvement throughout the ages, even starting in childhood. All antiheadache drugs, such as triptans, analgesics, ergots and opioids, along with the common combination substances currently on the pharmacological market are capable of inducing MOH. New data on specific clinical features and mean critical monthly dosages and mean critical monthly intake frequencies are now available. The only effective treatment concept is consequent withdrawal therapy. Data of prospective studies on relapse rates and predictors of relapse after successful withdrawal therapy are presented. PMID- 16370956 TI - Effects of common medications on cerebral vasospasm after subarachnoid haemorrhage. AB - Cerebral vasospasm is a common and serious complication of aneurysmal subarachnoid haemorrhage (SAH). At present, no consistently effective preventative and therapeutic measures are available, perhaps because of incomplete understanding of the pathogenesis of vasospasm. Experimental studies provide evidence that the incidence and severity of vasospasm after SAH can be modulated by drugs that affect neurotransmitter levels, intracellular signalling mechanisms, vascular smooth muscle function, inflammation and cellular proliferation, and the concentration of 'spasmogenic' factors. Preliminary clinical studies indicate that some illicit drugs and common prescription medications can have similar effects in humans. Recognition of these pharmacological effects is important because medications that can worsen or alleviate vasospasm are frequently administered to SAH patients to treat coincident medical problems. PMID- 16370957 TI - Coagulopathy and the role of recombinant human activated protein C in sepsis and following polytrauma. AB - Recombinant human activated protein C (rhAPC) also known as drotrecogin alfa (activated) has known antithrombotic, anti-inflammatory, and profibrinolytic properties in severe sepsis. Treatment with rhAPC (Xigris) has been shown to reduce mortality in patients with severe sepsis. The lack of any trials of rhAPC in trauma patients means that a definitive recommendation regarding its use in the polytraumatised patient, in whom severe head trauma or other contraindications for the use of rhAPC have been excluded remains controversial at present. This article describes the current evidence of its efficacy and safety in severe sepsis with relation to surgery and trauma. PMID- 16370958 TI - Comparative cardiovascular safety of traditional nonsteroidal anti-inflammatory drugs. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used for their anti inflammatory and analgesic effects. Unfortunately, these drugs are not without toxicity, namely on the gastric mucosa, but also on the cardiovascular system. In this context, the marketing of the coxibs, a new series of NSAIDs that selectively inhibit COX-2, resulted in a large debate around their cardiovascular safety, because they may increase the incidence of myocardial infarction and stroke. The recent suspension of a large, randomised, controlled trial comparing celecoxib, naproxen and placebo in Alzheimer patients (the ADAPT trial) because of an apparent elevated cardiovascular risk in the naproxen group revived the debate on the cardiovascular safety of these drugs, but this time with special emphasis on the effect of traditional nonselective NSAIDs (tNSAIDs). In this paper that reviews and discusses the cardiovascular safety profile of tNSAIDs, essentially naproxen and ibuprofen in view of the most recent experimental and clinical data, the authors note that the published data are quite discordant and one cannot conclude that there is clear evidence to support a cardiovascular hazard from the administration of naproxen or non-naproxen NSAIDs unless additional information is provided. In addition, the results of retrospective case-control studies have to be interpreted very carefully because of the risk of confounding factors that are not always taken into account when subjects were classified either as cases or controls. Thus, in the absence of clear cut data, physicians will have to use traditional NSAIDs (or coxibs) in patients with a high cardiovascular risk on the basis of their common sense rather than on evidence-based medicine. For these patients, one should not forget that an inadequate long-term control of cardiovascular risk factors such as a hypertension, dyslipidaemia, diabetes, smoking and weight excess is more deleterious in terms of cardiovascular mortality than the administration of NSAIDs itself. PMID- 16370959 TI - Drug-induced glomerulopathies. AB - Normal renal function depends upon an intact glomerular apparatus. Many drugs and chemicals are capable of damaging the glomerulus, causing its increased permeability to large molecules. Glomerular lesions are usually responsible for proteinuria and the nephrotic syndrome. This also holds true for the drug-induced glomerulopathies, of which membranous glomerulo-nephritis is the most frequent type of lesion encountered. Apart from this, several cases of different glomerular changes such as focal segmental glomerulosclerosis and crescentic glomerulonephritis have also been reported. The drug-induced glomerulopathies are probably immune mediated. This is, for instance, reflected in the fact that patients with drug-induced nephritic syndrome frequently have the HLA-B8 and DR3 antigens. In depth information is provided for the previously mentioned disorders. PMID- 16370960 TI - The safety and efficacy of antithyroid drugs. AB - Thionamides, selective inhibitors of thyroid peroxidase-mediated iodination by tyrosine residues in thyroglobulin, have been effectively used in the treatment of hyperthyroidism. The choices for initial treatment of patients with Graves' disease differ in various countries, and many physicians around the world prefer to administer thionamide drugs as the first choice of treatment for patients with hyperthyroidism. Although some thyroidologists more often consider radioiodine to be the treatment of choice because of its safety and ease of administration, thionamides remain the mainstay of treatment in thyrotoxic children and adolescents and in hyperthyroid women during pregnancy, postpartum period and lactation. A recent study with continuous thionamide treatment for patients with Graves' disease shows its efficacy, safety and cost-benefit properties. Further studies of the effectiveness of continuous thionamide therapy in patients with thyrotoxicosis need to be designed and implemented to determine indications for such therapy in children, adolescents and adults with diffuse toxic goiter, in particular, in those who have had recurrence of hyperthyroidism after discontinuation of one complete course of treatment. PMID- 16370961 TI - Is there a link between oestrogen therapy and gallbladder disease? AB - Cholelithiasis is the most common form of benign gallbladder disease that results in major heath expenditure. Female sex hormones are causally related to cholesterol gallstone disease, which are more common in women than in men. The risk of development of cholelithiasis is further enhanced by the use of exogenous female sex hormones and by pregnancy. Oestrogens are used in oral contraceptives and in hormone replacement therapy (HRT). Oral contraceptives do not pose a greater risk for gallbladder disease. The findings from two randomised, controlled trials, the Heart and Oestrogen/Progestin Replacement Study and the Women's Health Initiative postmenopausal hormone trial, unequivocally confirm that oral oestrogen use in postmenopausal women is causally associated with gallbladder disease, and the magnitude of the effect is not influenced greatly by the presence or absence of progestins. A cautious approach should be observed when prescribing HRT. Women must be informed about the effect of oestrogen use on increased risk of benign gallbladder disease. HRT should be used in the lowest possible dose for the shortest possible time. Women harbouring asymptomatic gallstones should not receive oestrogens because of the possibility of developing cholecystitis. PMID- 16370962 TI - Safety and side effects of the insulin analogues. AB - The analogue insulins were developed to more clearly mimic the basal and prandial components of insulin secretion for subjects with diabetes mellitus. Analogues are now widely used and have largely taken over from the conventional human recombinant insulins. It is important that these insulins are not only as effective as their predecessors, but are also safe and well-tolerated. In this manuscript, the authors review the adverse effects reported with analogue insulins and make a comparison with standard insulin treatments. PMID- 16370963 TI - Statin/fibrate combination in patients with metabolic syndrome or diabetes: evaluating the risks of pharmacokinetic drug interactions. AB - Patients with the metabolic syndrome and/or Type 2 diabetes mellitus continue to have a high risk of coronary heart disease (CHD) and progression of atherosclerotic lesions despite aggressive statin therapy. Although the National Cholesterol Education Programme Adult Treatment Panel III guidelines recommend the use of fibrates in combination with statins in patients at very high risk of CHD (e.g., patients at the low-density lipoprotein cholesterol target with high triglycerides and low high-density lipoprotein cholesterol, many physicians remain reluctant to use these combinations due to concerns of myotoxicity. Recently conducted metabolic and pharmacokinetic drug-drug interaction studies using gemfibrozil or fenofibrate in combination with five commonly used statins demonstrated a widely different drug interaction potential for these two fibrates. Gemfibrozil causes a 2- to 6-fold increase in statin area under the curve and increases the exposure to many recently approved drugs for the treatment of diabetes. Alternatively, fenofibrate does not adversely affect either the metabolism or pharmacokinetics of the statins studied. These pharmacokinetic differences appear to translate into less potential for interactions with fenofibrate/statin combination therapy compared to gemfibrozil/statin co-administration. The Action to Control Cardiovascular Risk in Diabetes (ACCORD) study in 10,000 patients with Type 2 diabetes mellitus is testing the efficacy and safety of fenofibrate/statin combination. PMID- 16370964 TI - The effect of antidepressants on glucose homeostasis and insulin sensitivity: synthesis and mechanisms. AB - OBJECTIVE: To synthesise results from investigations reporting on the effect of antidepressants on glucose-insulin homeostasis. METHOD: The authors conducted a MedLine search of all English language articles from 1966 to October 2005 using the keywords: bipolar disorder, major depressive disorder, diabetes mellitus, glucose homeostasis, and the name of each antidepressant that has been indicated for major depression in Canada and the US up to October 2005. The search was supplemented with a manual review of relevant references. Both preclinical and clinical investigations were reviewed. RESULTS: Some serotonergic antidepressants (e.g., fluoxetine) reduce hyperglycaemia, normalise glucose homeostasis and increase insulin sensitivity, whereas some noradrenergic antidepressants (e.g., desipramine) exert opposite effects. Dual-mechanism antidepressants (e.g., duloxetine and venlafaxine) do not appear to disrupt glucose homeostatic dynamics, whereas nonselective hydrazine monoamine oxidase inhibitors (e.g., phenelzine) are associated with hypoglycaemia and an increased glucose disposal rate. CONCLUSION: Some antidepressants exert a clinically significant effect on metabolism relevant to both therapeutic outcome and adverse events. PMID- 16370965 TI - Safety and efficacy of pediculicides for head lice. AB - Head lice infestation is a common and growing problem, primarily affecting school aged children. There are growing numbers of treatment failures due to the emergence of treatment-resistant lice to the popular over-the-counter products that have been used for the past several decades. Resistance has also decreased the efficacy of lindane, a prescription pediculicide that has been commonly used for several generations. Malathion, recently reintroduced in the US as a prescription pediculicide, has been associated with some treatment resistance depending upon its formulation. Other insecticidal treatments, such as ivermectin, will have to be developed further, given the limited options presently available for the treatment of head lice. Given the number of anecdotal and market-driven reported studies on head lice, assessment of topical lice therapies requires standardised in vitro testing. Based on concerns about safety and decreasing efficacy due to resistance, a reassessment of the general topic of pediculicides for head lice is warranted. PMID- 16370966 TI - Safety Pharmacology Society: 5th annual meeting. AB - This report summarises selected presentations delivered at the Safety Pharmacology Society annual meeting. Organ safety was covered, with cardiac QT liability being discussed most extensively. Of particular interest, was the lecture on beat-to-beat variability of cardiac repolarisation in anesthetised dogs with chronic atrioventricular block. This model, characterised by an electrophisyologically remodelled heart (replicating a human heart with reduced cardiac repolarisation reserve), was developed to assess the potential of drugs to cause unsafe QT prolongation that may spontaneously evolve into life threatening arrhythmias. An interesting case study illustrated the apparent failure of safety pharmacology studies to accurately predict clinical QT liability due to an underestimated plasma concentration for therapeutic efficacy at early nonclinical stage. An emerging attractive new technology, presented as a means to minimise drug failure to reach marketing stage (attrition) dealt with biological fingerprinting which consists in placing drug candidates in the context of chemical, pharmacological, toxicological and clinical liability spaces available for marketed, withdrawn or failed drugs, as well as reference compounds. The meeting was an excellent occasion for sharing knowledge and technologies among members of this young society actively engaged in promoting safety pharmacology, a novel research activity which now plays a pivotal role in the drug development process. PMID- 16370967 TI - Client participation and provider communication in family planning counseling: transcript analysis in Kenya. AB - To examine how much and in what ways clients participate in family planning consultations, and how providers influence their behavior, investigators analyzed transcripts of 178 counseling sessions with female clients in Kenya and developed coding guides that focus on client participation. The results show that providers dominate most counseling sessions, and clients rarely take an active role. The most common way for clients to participate is by volunteering additional information when they respond to providers' questions. Providers may encourage clients to play a more active role by building a sense of rapport, by relating contraceptive information specific to each client's personal situation, and by rewarding clients' attempts to participate. PMID- 16370968 TI - Social support and chronic fatigue syndrome. AB - The role of social support was examined in Chronic Fatigue Syndrome (CFS). Social support has been shown to affect illness outcome in medical disorders, likely due in part to communication between patient and support giver on illness-related concerns. Forty-one participants, 25 of whom had a primary support giver, completed the Beck Depression Inventory, the Perceived Stress Scale, the Profile of Mood States, and the Inventory of Socially Supportive Behaviors (ISSB); the primary support giver completed a modified version of the ISSB indicating the level of support he or she provided and a questionnaire assessing beliefs about CFS. Results indicated that there were no differences among individuals with CFS with or without support on measures of mood and perceived stress. Individuals with CFS and their support givers agreed on the amount of support offered, and extent of support was independent of beliefs concerning etiology. Exploratory analyses revealed that verbal emotional-cognitive support generally was more predictive of mental health than was tangible, less communicative support. The lack of positive effect of social support is discussed in relation to the degree of support offered, and implications for future research are discussed. PMID- 16370969 TI - Blood, vomit, and communication: the days and nights of an intern on call. AB - Internship year is the first year of a medical residency, and often the hardest due to an intern's amount of work and lack of experience. This research provides an evocative account of the internship context based on observations of an intern, who was shadowed during 2 of his nights on call. The author describes the fragmented, chaotic, and exhausting experiences of an intern. This descriptive account contrasts with current health communication literature, in that it recognizes a specific context rather than assuming a nondescript clinical setting. Conclusions suggest that the experiences of internship year offer little opportunity to develop the communication skills necessary for a physician's later clinical career. In addition, health communication researchers must strive to better understand medical training by more frequently entering the trenches in which interns work. Such research will serve to advance interpersonal communication within specific health care contexts. PMID- 16370970 TI - Prostate cancer's hegemonic masculinity in select print mass media depictions (1974-1995). AB - The meanings associated with prostate cancer were studied in contemporary mass print media. The study includes both manifest and latent content analysis of a period of approximately 2 decades, from 1974 to 1995. The manifest analysis revealed a primary emphasis on the importance of early detection. The latent analysis found that prostate cancer's presentation is gendered. Its description is embedded in themes related to masculinity, sexuality, competition, brotherhood, and machismo. This small, qualitative, and inductive study raises questions about the socially significant portrayal of the meanings of disease in the media, about the men who have been diagnosed with prostate cancer, have symptoms of prostate cancer, or about all men, because any man might at some time be diagnosed with prostate cancer. Stereotypical imaging could alienate men who either do not or do not want to fit into the stereotypical ideal as it is protrayed in the media. Such a portrayal also may have inplications for the potential willingness of men to engage in early detection, avail themselves of treatment, act preventatively, or become involved in lobbying for monies for research into the early prevention, detection, and treatment of prostate cancer. PMID- 16370971 TI - Maternal personality and health communication in the pediatric context. AB - Maternal personality and social skills were used to predict both maternal and pediatrician communication patterns during pediatric well-child examinations. Audiotape recordings of mother-pediatrician interaction were collected and analyzed from 78 mothers of children less than 2 years of age at 5 private practice pediatric offices; questionnaires were completed at home by the mothers. Canonical and zero-order correlations indicated that specific utterances made by both mothers and pediatricians can be predicted from the mother's personality (particularly with regard to openness to experience, extraversion, and neuroticism) and from the mother's social skills, providing support for a dynamic mutuality model of medical communication. Theoretical and applied implications for doctor training and development of parent education programs are discussed. PMID- 16370972 TI - The patient self-advocacy scale: measuring patient involvement in health care decision-making interactions. AB - Despite the fact that many individuals express a desire for more information and involvement in the health care process, it remains to be seen if they have adopted a more participative approach by becoming involved in decisions made about their health. Research indicates that, in actual practice, individuals are differentially willing or able to be active patients. AIDS patient activists are 1 group of individuals who have become more involved in their health care decision making. This study tests the reliability and validity of a measure of patient activism-the Patient Self-Advocacy Scale (PSAS)--designed to assess the dimensions of (a) increased illness and treatment education, (b) increased assertiveness in health care interactions, and (c) increased potential for nonadherence. Tests administered to 2 samples of participants (174 adults from an HIV-AIDS population and 21 8 adults from a general population) demonstrated that the PSAS was a reliable and valid measure of patient involvement in health care decision making. PMID- 16370973 TI - Communication of social support in computer-mediated groups for people with disabilities. AB - This study documented the types and extent of social support messages exchanged by persons with disabilities who participated in a computer-based support group. A modified version of Cutrona & Suhr's (1992) social support category system was used to code 1,472 support messages. The largest percentage of these messages offered emotional and informational support, whereas network support and tangible assistance were least frequently offered. It appeared that many of the support messages directly redressed limitations and challenges associated with disability related mobility, socialization, and self-care. Results are discussed in terms of the generalizability of existing category systems for coding support to this mediated context, the relative importance of different types of support in the communication of support group members, and the unique features of social support in mediated environments. The implications of this study for social support researchers, persons with disabilities, and human services professionals are also discussed. PMID- 16370974 TI - Listening to women's narratives of breast cancer treatment: a feminist approach to patient satisfaction with physician-patient communication. AB - Traditional health communication research often has ignored sex and gender and has employed a quantitative biomedical perspective to predict behavior. In contrast, this study analyzed women's narratives of their breast cancer treatment to uncover conceptualizations of patient satisfaction with physician-patient communication. In their unfolding (nonlinear) narratives, patients viewed satisfaction as a negotiation process with physicians in which themes of respect, caring, and reassurance of expertise were prominent. Two root themes (dialogic approach to power and contextualization) acted as underlying dynamics or tensions throughout their narratives. Patients' ways of knowing and preferences for feminine communication styles influenced perceptions of physician-patient communication satisfaction. PMID- 16370976 TI - The present and future of health communication scholarship: overlooked opportunities. PMID- 16370975 TI - Explaining illness to children: advancing theory and research by determining message content. AB - Numerous scholars have suggested the importance of explaining illness to children and empirical efforts have been made to determine effective illness explanation strategies. However, the studies have been few and provide little advancement of theory and practical advice for health practitioners. Much, if not all, of the theory and research in this area focuses on explaining the etiology and biology of illness to children. This practice, as argued here, may be the central factor inhibiting theory, research, and understanding of effective illness explanation to children. Specifically, children may find this knowledge of little use and may prefer explanations that adhere to psychosocial concerns. As such, suggestions for future research are offered. PMID- 16370977 TI - Punishing versus reinforcing strategies of drug discontinuance: effect of persuaders' drug use. AB - This study tests and extends inconsistent nurturing as control theory (Le Poire, 1992,1995) by exploring the use of reinforcing and punishing drug discontinuance strategies based on the drug-use status of the functional/persuading partners (past abuse, current abuse, current use, and nonuse). All partners were inconsistent in their use of reinforcement and punishment of substance abuse, with past abusers punishing the substance abuse most before they labeled the drug use as problematic, and current users and nonusers punishing the substance abuse the most following the labeling and in the postfrustration period. Additionally, current abusers were the most reinforcing of alternative behavior during every time period, a strategy that was most highly related to reduction in relapse. Furthermore, nonusers utilized the most indulgence and antidrink strategies, that are in opposition based on their reinforcing and punishing natures. Past abusers were rated as most persuasively effective by their partners, whereas nonusers were evaluated as the least persuasively effective. Finally, drug use was related to the mental health of the persuading partner, in that current abusers and nonusers were significantly more depressed and anxious than past abusers or current users. PMID- 16370978 TI - Factors influencing caregiver-care receiver communication and its impact on the well-being older care receivers. PMID- 16370979 TI - Nurturing communication by health professionals toward patients: a communication accommodation theory approach. AB - This study explores the role of nurturing communication in distinguishing interpersonal and intergroup interactions between health professionals and patients, from the perspective of communication accommodation theory (CAT). Participants (47 men and 87 women) rated videotapes of actual hospital consultations on 12 goal and 16 strategy items derived from CAT. Health professionals in interpersonal interactions were perceived to pay more attention to relationship and emotional needs and to use more nurturant discourse management and emotional expression. These results point the way toward elucidating the perceived optimal balance in accommodative behavior, both group based and interpersonal, in these contexts, and they highlight the importance of nurturant communication to this process. PMID- 16370980 TI - "You're out of your mind!": Humor as a face-saving strategy during neuropsychological examinations. AB - This article presents a sociolinguistic analysis of humor as a face-saving device in a memory clinic. Data for this article were transcripts of audiotaped clinical examinations between 4 clinicians and 17 patients, conducted at the Memory and Alzheimer's Clinic at the University of California, San Francisco. The study focused on the functions of humor and the complexity involved in examining those functions. Four aspects of humor were examined: (a) who initiates humor, (b) what topics the humor is based on, (c) what the function of the humor is, and (d) who the focus of the humor is. Results indicated that dementia patients initiated a greater number of humor exchanges than did clinicians or third-party observers. In particular, patients initiated a greater number of a specific kind, called dominant humor, with which the initiator controls the interaction. In spite of the power differential between patients and clinicians, dementia patients asserted some dominance over a face-threatening situation. These findings imply the need for further research on nurturing, as well as communicative training of family and caregivers of dementia patients to enhance successful communication and thus successful life experiences for these patients. PMID- 16370981 TI - The language of mentoring in a health care environment. AB - In this article, 4 models were tested on the relations among mentoring, job expectations, job reward value, job involvement, and organizational commitment of hospital nurses. In the model with the best fit to the data, mentoring and job expectations were positively related to job reward value. Organizational commitment and job involvement were increased by job reward value. A direct effect on organizational commitment was found for mentor assistance in learning skills. Specific methods used by mentors to convey supportive communication to their proteges included encouraging them to do their best, helping them gain additional training, providing support and encouragement when others criticize, helping deal with jealousy, and providing proteges with needed information. Nurse mentors were admired for standing up to administration and for challenging unfair decisions, thoughtless actions, and lack of respect on the part of administrators, doctors, and patients. Nurses indicated that working hard and not having anyone notice, being frequently criticized and rarely rewarded, and being exposed to serious diseases were particularly challenging and stressful aspects of nursing. Implications of this study for nurses and those working in health care environments are that mentoring and organizational commitment may be avenues for increasing the degree to which nurses feel that their job is rewarding. PMID- 16370982 TI - Interviews with elderly patients about side effects. AB - Fifty older persons were interviewed about their medicines, side effects, and their responses to statements describing side effects. Participants were taking nearly 4 drugs each and knew what those medicines were and why they were taking them. Participants knew much less about the potential side effects and almost nothing about potential drug interactions. They wanted to know all the side effects, and indicated that if their doctors told them about all the side effects, they would still follow the doctors' prescription. However, fewer than 1 in 3 of their doctors were doing so. Responses to hypothetical statements about a high blood pressure medication showed that positive statements were more reassuring than negative statements. Responses to statements about side effects were more driven by the nature of the side effect than by its likelihood. Overall, participants did not find side-effect information strongly discouraging. PMID- 16370983 TI - Processing messages about disease detection and health promotion behaviors: the effects of anxiety. AB - A study was conducted to examine the effects of anxiety on the processing of messages that encourage the performance of disease detection and health promotion behaviors. It was hypothesized that under high-anxiety conditions messages about health promotion behaviors would be processed more than messages about disease detection behaviors and that this effect would reverse under lower levels of anxiety. To test this hypothesis, the participants were required to read information designed either to increase or to decrease anxiety about health. Following the anxiety manipulation, participants received a strong or weak message promoting the performance of either health promotion or disease detection behaviors. Then participants were required to indicate their attitudes about the behavior, their cognitive responses to the message, and their recall of the message. The results support the hypothesis. PMID- 16370984 TI - Testing the influence of the health belief model and a television program on nutrition behavior. AB - This study proposed to identify relevant factors of the health belief model (HBM) that provide motivation for people to engage in healthy dietary behavior. The impact of a TV program was also assessed using a longitudinal study with 300 participants and measuring the influence of path coefficients in the HBM that predicted salience, motivation, and healthy eating practices. Findings suggested that nutrition behavior was influenced by susceptibility and efficacy mediated through health motivation and salience. Program viewing boosted salience regardless of age, education, or household size by significantly increasing viewers' confidence in their nutrition knowledge base. PMID- 16370985 TI - The perception of risk messages regarding electromagnetic fields: extending the extended parallel process model to an unknown risk. AB - The Extended Parallel Process Model (EPPM) was developed as a model to assist in the development of effective risk communication messages, specifically messages that elicit adaptive behavioral responses. It has shown to be effective in several settings invoking clearly delineated dangers (e.g., safety belt usage, condom usage).Unfortunately, communicating risk messages is not always so straightforward. One increasing concern in the risk communication field is the controversy over electromagnetic fields (EMFs) and the uncertain hazards they present to individuals. The purpose of this study is to test the EPPM with this unknown risk and to explore which type of risk message may motivate adaptive behavioral responses. In accordance with the EPPM model, 251 participants received either a low- or high-threat risk message and a list of control measures they could use to reduce their exposure to EMFs. Results suggest that the EPPM model can be extended to an unknown risk. PMID- 16370986 TI - The development and partial assessment of the medical communication competence scale. AB - The purpose of this research was to develop and partially assess a self-report scale for measuring doctors' and patients' perceptions of self-communication and other communication competence during a medical interview. Previous research into the components of communication competence and medical discourse were used to develop the Medical Communication Competence Scale (MCCS). It was hypothesized that the items of the MCCS would form four clusters: information giving, information seeking, information verifying, and socioemotional communication. The cluster analysis results provided support for the hypothesis. Results of several other analyses provided additional support for the validity of the MCCS. PMID- 16370987 TI - The many meanings of uncertainty in illness: toward a systematic accounting. AB - Uncertainty is widely believed to be a central feature in illness experiences. Moreover, communication is thought to be essential to the construction, management, and resolution of uncertainty. Not surprisingly, however, there are substantial variations in conceptions and analyses of this focal construct and its relation to communication. In this article, we first argue that understandings of the role of communication in health and illness can be improved substantially--thereby enhancing both theory development and practical applications of health communication theory--by recognizing and reconciling sources of variation in conceptions of uncertainty and by synthesizing various specific conceptions. We then review individual-psychological models. linguistic and discourse analyses, and sociocultural and historical perspectives on uncertainty in illness Following the review, we present a framework that synthesizes many conceptions of uncertainty We close with a discussion of 5 challenges and opportunities for research and application. PMID- 16370988 TI - Who learns preventive health care information from where: cross-channel and repertoire comparisons. AB - We studied from which information channels individuals reported learning the most information about preventive health care, how those channels correlated with one another, and how well they were predicted by demographics and health orientations. A probability sample of 1,963 adults from 8 midwestern communities were interviewed from late 1994 to early 1995. Respondents reported learning different amounts of preventive health information from different channels, and a mix in levels of learning was found across channels. Television news and information rated unexpectedly high across the population studied. An exploratory factor analysis indicated a clear grouping or repertoire consisting of television channels, and for magazines and newspapers, but also a distinct personal media repertoire involving a mix of health professionals, family and friends, books, educational materials, and computers. Demographics did better at predicting learning from traditional print media, but personal health orientations were more effective predictors of personal media; television was less well predicted by either. PMID- 16370989 TI - A preliminary analysis of memorable support and nonsupport messages received by nurses in acute care settings. AB - This study presents a preliminary analysis of recalled messages for support and nonsupport received by nurses in a midsize, midwestern hospital. In interviews nurses recalled specific incidents of support and nonsupport they had experienced in the workplace. The structural and functional characteristics of these memorable messages of support and nonsupport were then independently coded by trained coders within the following categories: (a) the structure and form of the message, (b) the context and timing of the message, (c) the relationship between the source and receiver, and (d) the content of the message and the message effects. The results of chi-square analyses and McNemar tests are presented. The results suggest that in the specific categorical areas examined, support and nonsupport messages exhibit a relatively high degree of similarity within participants. That is, participants appeared to use a cognitive template for classifying communication acts as supportive and nonsupportive. Specific between participant message similarities and differences are also enumerated and explained. PMID- 16370990 TI - In search of cultural sensitivity and inclusiveness: communication strategies used in rural HIV prevention campaigns designed for African Americans. AB - Recently, rates of HIV and AIDS are rapidly and disproportionately increasing among minority communities in the rural South. Culturally specific health communication about HIV and inclusion of minority voices in the administration and implementation of HIV programs have been found to be the most effective methods for prevention. The purpose of this discussion is to examine these health communication strategies in HIV prevention programs designed for African American communities in rural Alabama. Effective, culturally sensitive, and inclusive prevention efforts documented in health communication literature are identified, and the use of these efforts through a case study of rural Alabama's minority focused HIV prevention programs is examined The research reveals that, although the state is making use of culturally tailored communication strategies, educators continue to encounter problems connecting with and involving target populations. Reasons for these problems and recommendations for changes are discussed. PMID- 16370991 TI - Anti-smoking socialization: relationship to parent and child smoking status. AB - Anti-smoking socialization is defined as the transmission of knowledge, attitudes, and skills that prepare children to resist smoking. Three types of anti-smoking socialization are studied: parents' communication with children regarding no-smoking rules at home, health risks associated with smoking, and the disciplinary consequences of smoking. Results from a survey of children in 3rd through 8th grades (N = 937) indicate that (a) children from households where one or both parents smoke perceive less anti-smoking socialization than their peers from nonsmoking households; (b) anti-smoking socialization is positively associated with authoritative parenting; (c) children have significantly lower rates of smoking intention and initiation when parents engage in anti-smoking socialization, even if parents currently smoke. The findings offer an encouraging message to all parents Communicating anti-smoking messages may effectively discourage children from smoking. PMID- 16370994 TI - 'But basically you're feeling well, are you?': tag questions in medical consultations. AB - Tag questions have traditionally been discussed as linguistic strategies used by nonassertive speakers. This article reports the findings of a qualitative study investigating the use of modal and affective tag questions by 3 Australian female general practitioners. Analysis of 29 audiotaped consultations revealed that tag questions were used as both control and involvement strategies. They were found to be employed by doctors for their potential to elicit information from patients, to summarize and confirm information, and to express empathy and provide positive feedback. The implications of these findings are discussed in the light of general patterns of doctor-patient communication as well as more efficient clinical interaction. PMID- 16370995 TI - Relational control patterns in physician-patient clinical encounters: continuing the conversation. AB - The purpose of this study is to continue the application of the relational communication approach for describing physician-patient control patterns and provide initial exploration into the implication of these control behaviors on patient satisfaction and compliance. Videotaped family medicine clinical visits were analyzed with Rogers and Farace's (1975) Relational Communication Control Coding Scheme. In addition, telephone surveys were conducted with patients 2 to 3 weeks after the clinical visit to assess levels of patient satisfaction and compliance. Transactional results indicated physicians manifested more control submission, whereas patients engaged in greater control dominance. The resulting outcomes of control behaviors showed an increase in patient compliance when physicians exhibited less control assertiveness and patients showed less control submission. An increase in patient satisfaction was found when physicians showed less control dominance. PMID- 16370996 TI - Bonding and cracking: the role of informal, interpersonal networks in health care decision making. AB - This study examined the role and impact of informal, interpersonal networks in health care decision making. Questioning the traditional approach to studying health communication as it is situated within institutions (e.g., Burgoon, 1992; Nussbaum, 1989; Pettegrew & Logan, 1987), this ethnographic study ventured into the lived experiences of women in a mothers' and toddlers' playgroup as they discussed their own, their family members', and their friends' health experiences. Their conversations revolved around such health-related issues as pregnancy and delivery, physicians and hospitals, breastfeeding, illnesses and accidents, and diet and nutrition. The conversations were found to serve not necessarily opposing but distinctive functions including a rather practical purpose of "cracking the code" of institutional practices, as well as a "bonding" function evidenced through stories or narratives. The conversations are not only exchanges of information but also narratives through which shared experiences are created and maintained. In addition to these narrative functions, examination of these conversations reveals how diagnoses and remedies from the institutions are discussed and decisions are made. The conclusion offers ideas for how agencies and institutions can utilize the findings of this particular research venture. PMID- 16370997 TI - Bearing the burden or baring the soul: physicians' self-disclosure and boundary management regarding medical mistakes. AB - Within a boundary management framework, this study explored how physicians manage self-disclosure regarding medical mistakes amidst boundary constraints imposed by risk management, legal mandate, and the medical culture. Descriptive statistics from questionnaires and exemplars from accompanying narratives showed that the 39 internists and family physicians in this study chose to control their own boundaries by revealing errors most often to other physicians to facilitate learning. Although risk management and the medical culture do not appear to deter physicians from disclosing errors at a superficial level, physicians maintain tight personal boundaries at the emotional level. Perhaps if physicians could disclose errors at the emotional level, their mental energies could be more positively channeled to patients' needs, resulting in improved patient care. PMID- 16370998 TI - Current issues in communication research. PMID- 16371000 TI - Studying health communication: an agenda for the future. PMID- 16370999 TI - Setting the agenda for health communication research and development: scholarship that can make a difference. PMID- 16371001 TI - Improving citizens' health competencies. PMID- 16371002 TI - Directions for research within health communication. PMID- 16371003 TI - Emotion, reason, and communication in coping with cancer. AB - Researchers have sought to identify which coping styles are most conducive to recovery from cancer and other serious illnesses. The results have been equivocal. It appears that what helps one patient does not necessarily help another. This article proposes a model explaining how emotional, cognitive, and environmental conditions influence the styles of coping used by cancer patients and the likelihood that particular styles will prove helpful to these patients. The model is based on the premise that coping does not occur in isolation from the emotional makeup, cognitive predispositions, and social environment of the patient. It describes how these processes interact to influence the protective effect of various coping styles. PMID- 16371004 TI - The influence of a companion on the doctor-elderly patient interaction. AB - Based on findings that elderly patients brought companions to their medical appointments more often than middle-aged patients, the influence of the companion was examined. Twelve of 21 patients 60 to 85 years old brought companions. There was no significant difference in length of interaction for patients with and without companions, indicating that companions, by speaking, took time away from patients. Doctors directed fewer comments to companions than companions directed to doctors, indicating that companions responded or initiated comments when doctors were not addressing them. Companions played three roles: watchdog, significant other, and surrogate patient. PMID- 16371006 TI - Unexpected similarity between the cytosolic West Nile virus NS3 and the secretory furin-like serine proteinases. AB - Many viral proteins undergo proteolytic processing events that are required for virus infection and virion assembly. In this issue of Biochemical Journal, Strongin and co-workers report that the NS3 protease from West Nile virus unexpectedly cleaves certain substrates at pairs of basic residues, a specificity that resembles that of the furin-like PCs (proprotein convertases). This led to the demonstration that furin/PC inhibitors containing poly(D-arginine) are also potent inhibitors of NS3, and that anthrax toxin protective antigen and myelin basic protein are potential NS3 substrates. Structural modelling based on Dengue virus NS3 provided a possible rationale for the observed cleavage specificity of West Nile virus NS3. PMID- 16371007 TI - Direct evidence for S-nitrosation of mitochondrial complex I. AB - NO* (nitric oxide) is a pleiotropic signalling molecule, with many of its effects on cell function being elicited at the level of the mitochondrion. In addition to the well-characterized binding of NO* to the Cu(B)/haem-a3 site in mitochondrial complex IV, it has been proposed by several laboratories that complex I can be inhibited by S-nitrosation of a cysteine. However, direct molecular evidence for this is lacking. In this investigation we have combined separation techniques for complex I (blue-native gel electrophoresis, Superose 6 column chromatography) with sensitive detection methods for S-nitrosothiols (chemiluminescence, biotin switch assay), to show that the 75 kDa subunit of complex I is S-nitrosated in mitochondria treated with S-nitrosoglutathione (10 microM-1 mM). The stoichiometry of S-nitrosation was 7:1 (i.e. 7 mol of S-nitrosothiols per mol of complex I) and this resulted in significant inhibition of the complex. Furthermore, S-nitrosothiols were detected in mitochondria isolated from hearts subjected to ischaemic preconditioning. The implications of these results for the physiological regulation of respiration, for reactive oxygen species generation and for a potential role of S-nitrosation in cardioprotection are discussed. PMID- 16371008 TI - Extracellular N-acetylaspartate depletion in traumatic brain injury. AB - N-Acetylaspartate (NAA) is almost exclusively localized in neurons in the adult brain and is present in high concentration in the CNS. It can be measured by proton magnetic resonance spectroscopy and is seen as a marker of neuronal damage and death. NMR spectroscopy and animal models have shown NAA depletion to occur in various types of chronic and acute brain injury. We investigated 19 patients with traumatic brain injury (TBI). Microdialysis was utilized to recover NAA, lactate, pyruvate, glycerol and glutamate, at 12-h intervals. These markers were correlated with survival and a 6-month Glasgow Outcome Score. Eleven patients died and eight survived. A linear mixed model analysis showed a significant effect of outcome and of the interaction between time of injury and outcome on NAA levels (p = 0.009 and p = 0.004, respectively). Overall, extracellular NAA was 34% lower in non-survivors. A significant non-recoverable fall was observed in this group from day 4 onwards, with a concomitant rise in lactate-pyruvate ratio and glycerol. These results suggest that mitochondrial dysfunction is a significant contributor to poor outcome following TBI and propose extracellular NAA as a potential marker for monitoring interventions aimed at preserving mitochondrial function. PMID- 16371009 TI - Contribution of conserved polar glutamine, asparagine and threonine residues and glycosylation to agonist action at human P2X1 receptors for ATP. AB - The role of conserved polar glutamine, asparagine and threonine residues in the large extracellular loop, and glycosylation, to agonist action at human P2X1 receptors was tested by generating alanine substitution mutants. For the majority of mutants (Q56A, Q95A, T104A, T109A, Q112A, Q114A, T146A, N153A, T158A, N184A, N191A, N242A, N300A) alanine substitution had no effect on ATP potency. The mutants Q95A, Q112A, Q114A and T158A showed changes in efficacy for the partial agonists BzATP and Ap5A, suggesting that these polar residues may contribute to the gating of the channel. The mutants T186A, N204A and N290A had six-, three- and 60-fold decreases in ATP potency, respectively. For T186A and N290A, the partial agonists BzATP and Ap5A were no longer agonists but still bind to the receptor as shown by the ability to modulate the response to co-applied ATP. N153, N184 and N242 are glycosylated in the endoplasmic reticulum and N300 acquires complex glycosylation in the golgi. These results aid in refining a model for ATP binding at the P2X1 receptor where the residues F185T186, and the conserved triplet N290F291R292, are likely to play a role in ATP action at the receptor. PMID- 16371010 TI - Loss of metabotropic glutamate receptor-mediated regulation of glutamate transport in chemically activated astrocytes in a rat model of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by a selective loss of motor neurones accompanied by intense gliosis in lesioned areas of the brain and spinal cord. Glutamate-mediated excitotoxicity resulting from impaired astroglial uptake constitutes one of the current pathophysiological hypotheses explaining the progression of the disease. In this study, we examined the regulation of glutamate transporters by type 5 metabotropic glutamate receptor (mGluR5) in activated astrocytes derived from transgenic rats carrying an ALS-related mutated human superoxide dismutase 1 (hSOD1(G93A)) transgene. Cells from transgenic animals and wild-type littermates showed similar expression of glutamate-aspartate transporter and glutamate transporter 1 (GLT-1) after in vitro activation, whereas cells carrying the hSOD1 mutation showed a three-fold higher expression of functional mGluR5, as observed in the spinal cord of end stage animals. In cells from wild-type animals, (S)-3,5-dihydroxyphenylglycine (DHPG) caused an immediate protein kinase C (PKC)-dependent up-regulation of aspartate uptake that reflected the activation of GLT-1. Although this effect was mimicked in both cultures by direct activation of PKC using phorbol myristate acetate, DHPG failed to up-regulate aspartate uptake in cells derived from the transgenic rats. The failure of activated mGluR5 to increase glutamate uptake in astrocytes derived from this animal model of ALS supports the theory of glutamate excitotoxicity in the pathogenesis of the disease. PMID- 16371011 TI - The alternative splicing of tau exon 10 and its regulatory proteins CLK2 and TRA2 BETA1 changes in sporadic Alzheimer's disease. AB - Pathological inclusions containing fibrillar aggregates of hyperphosphorylated tau protein are a characteristic feature in tauopathies, which include Alzheimer's disease (AD). Tau is a microtubule-associated protein whose transcript undergoes alternative splicing in the brain. Exon 10 encodes one of four microtubule-binding repeats. Exon 10 inclusion gives rise to tau protein isoforms containing four microtubule-binding repeats (4R) whereas exclusion leads to isoforms containing only three repeats (3R). The ratio between 3R and 4R isoforms is tightly controlled via alternative splicing in the human adult nervous system and distortion of this balance results in neurodegeneration. Previous studies showed that several splicing regulators, among them hTRA2-beta1 and CLK2, regulate exon 10 alternative splicing. Like most splicing factors, htra2-beta and clk2 pre-mRNAs are regulated by alternative splicing. Here, we investigated whether human postmortem brain tissue of AD patients reveal differences in alternative splicing patterns of the tau, htra2-beta, presenilin 2 and clk2 genes when compared with age-matched controls. We found that the splicing patterns of all four genes are altered in affected brain areas of sporadic AD patients. In these affected areas, the amount of mRNAs of tau isoforms including exon 10, the htra2-beta1 isoform and an inactive form of clk2 are significantly increased. These findings suggest that a misregulation of alternative splicing seems to contribute to sporadic AD. PMID- 16371012 TI - Renal medullary carcinoma, a rare cause of haematuria in sickle cell trait. PMID- 16371013 TI - Multicolour banding provides a detailed characterisation of structural abnormalities of chromosome 1 in Burkitt lymphoma. PMID- 16371014 TI - Spectrum of infection, risk and recommendations for prophylaxis and screening among patients with lymphoproliferative disorders treated with alemtuzumab*. AB - There is an increasing use of monoclonal antibodies in the treatment of haematological malignancies. Alemtuzumab (Campath-1H; Ilex Pharmaceuticals, San Antonio, TX, USA) is a monoclonal antibody reactive with the CD52 antigen used as first and second line therapy for two types of lymphoproliferative disorders: chronic lymphocytic leukaemia (CLL), and T-cell lymphomas [both peripheral (PTCL) and cutaneous (CTCL)]. With alemtuzumab therapy, viral, bacterial and fungal infectious complications are frequent, and may be life threatening. An understanding of the patients at highest risk and duration of risk are important in developing recommendations for empirical management, antimicrobial prophylaxis and targeted surveillance. This review discusses the infection risks associated with these lymphoproliferative disorders and their treatment, and provide detailed recommendations for screening and prophylaxis. PMID- 16371015 TI - Managing the risk of transmission of variant Creutzfeldt Jakob disease by blood products. AB - Whereas plasma-derived clotting factor concentrates now have a very good safety record for not being infectious for lipid enveloped viruses, concern has arisen about the possibility that prion diseases might be transmitted by blood products. There is epidemiological evidence that classical sporadic Creutzfeld Jakob disease (CJD) is not transmitted by blood transfusion. There is now good evidence that the abnormal prion associated with variant CJD can be transmitted by transfusion of fresh blood components and infect recipients. To reduce the risk of the pathological prion in the UK infecting recipients of clotting factor concentrates, these are now only manufactured from imported plasma collected from countries where there has not been bovine spongiform encephalopathy (BSE) in cattle and the risk of variant CJD in the population is, therefore, considered negligible. The safety of these concentrates is also enhanced because prion protein is, to an appreciable extent, excluded by the manufacturing process from the final product. To help reduce the chance of prion transmission by fresh blood products, donations are leucodepleted, there is increasing use of imported fresh frozen plasma (especially for treating children) and potential donors, who have been recipients of blood since 1980 (the beginning of the BSE epidemic in cattle) are deferred. PMID- 16371016 TI - Short-term mortality of bacteraemia in elderly patients with haematological malignancies. AB - Bacterial infections are important complications in patients with haematological malignancies. We compared the outcome of bacteraemia among elderly and younger patients with haematological malignancies, and evaluated the impact of comorbidity on this association using population-based registries from 1992 to 2002. Among 358 patients with an incident haematological malignancy and an episode of bacteraemia, 207 (58%) were older than 60 years and 37 (10%) older than 80 years. The 7-d mortality was 10% among patients younger than 60 years, 21% among patients aged 60-79 years, and 27% for patients older than 80 years. When compared with patients younger than 60 years, the adjusted mortality rate ratios (MRRs) were 1.9 [95% confidence interval (CI): 0.9-3.8] for patients aged 60-79 years and 1.6 (95% CI: 0.6-4.2) for patients older than 80 years. The 30-d mortality was 23% among patients younger than 60 years of age, 35% among patients aged 60-79 years, and 54% among patients 80 years or older. Adjusted MRRs were 1.7 (95% CI: 1.1-2.7) and 2.3 (95% CI: 1.2-4.3), respectively. We found that increasing age was associated with increased mortality from bacteraemia in patients with haematological malignancies. An increased burden of comorbidity among the elderly did not explain this association. PMID- 16371017 TI - Simultaneous cytokine analysis by cytometric bead array for the detection of leukaemia-reactive T cells in patients with chronic myeloid leukaemia. AB - In order to detect T cells against several chronic myeloid leukaemia (CML) associated antigens we used: (i) a novel T-cell assay [cytometric bead array (CBA)]; (ii) gamma-interferon enzyme-linked immunoSPOT (gamma-IFN-ELISpot); and (iii) tetramer staining in peripheral blood from CML patients. Peptide-specific cytokine release was detected by CBA in some patients, whereas standard gamma-IFN ELISpot and tetramer staining were negative in the vast majority of cases. In CBA, peptide-specific cytokine release was predominantly tumour necrosis factor alpha, raising questions about the responding cells and their functional status. CBA appears to be a new useful tool for the detection of leukaemia-reactive T cells. PMID- 16371018 TI - Graft failure following reduced-intensity cord blood transplantation for adult patients. AB - We reviewed the medical records of 123 adult reduced-intensity cord blood transplantation (RI-CBT) recipients to investigate the clinical features of graft failure after RI-CBT. Nine (7.3%) had graft failure, and were classified as graft rejection rather than primary graft failure; they showed peripheral cytopenia with complete loss of donor-type haematopoiesis, implying destruction of donor cells by immunological mechanisms rather than poor graft function. Three of them died of bacterial or fungal infection during neutropenia. Two recovered autologous haematopoiesis. The remaining four patients underwent a second RI-CBT and developed severe regimen-related toxicities. One died of pneumonia on day 8, and the other three achieved engraftment. Two of them died of transplant-related mortality, and the other survived without disease progression for 9.0 months after the second RI-CBT. In total, seven of the nine patients with graft failure died. The median survival of those with graft failure was 3.8 months (range, 0.9 15.4). Graft failure is a serious complication of RI-CBT. As host T cells cannot completely be eliminated by reduced-intensity preparative regimens, we need to be aware of the difficulty in differentiating graft rejection from other causes of graft failure following RI-CBT. Further studies are warranted to establish optimal diagnostic and treatment strategies. PMID- 16371019 TI - Randomised comparison of two B-cell purging protocols for patients with B-cell non-Hodgkin lymphoma: in vivo purging with rituximab versus ex vivo purging with CliniMACS CD34 cell enrichment device. AB - We investigated the feasibility, safety and efficacy of two B-cell purging methods in patients with CD20+ non-Hodgkin lymphoma (NHL) receiving autologous stsem cell transplantation. Myeloid and immune recoveries between the methods were compared. Twenty-seven patients were randomised to either in vivo purging with rituximab or ex vivo purging by CD34+ cell selection. Both purging methods were efficient at eliminating B-cells in infusates. When compared with in vivo purging, ex vivo purging was associated with CD34+ cell loss and delayed median neutrophil (10 d vs. 11 d) and platelet (12.5 d vs. 17 d) count recoveries. Lymphocyte recovery was similar in both groups, but immunoglobulin recovery was delayed after in vivo purging. Late-infectious complications were few in both arms. At a median follow-up of 27 months, 2-year probabilities of event-free survival (EFS) rates were 81% for in vivo purging and 76% for ex vivo purging (P = 0.66). When compared with 53 unpurged patients, all 27 purged patients had improved 3-year probabilities of overall survival (89% vs. 70%, P = 0.014) and a trend for improved EFS (78% vs. 57%, P = 0.075). In conclusion, although both purging methods were feasible and safe, rituximab purging was superior as it did not impair CD34+ cell mobilisation and was associated with faster myeloid recovery. Further studies are needed to determine whether rituximab purging is more effective than the use of unpurged autografts. PMID- 16371020 TI - Possible involvement of allogeneic antigens recognised by donor-derived CD4 cytotoxic T cells in selective GVL effects after stem cell transplantation of patients with haematological malignancy. AB - Cytotoxic T lymphocyte (CTL) lines specific for allogeneic antigens were generated by in vitro stimulation of donor-derived peripheral blood mononuclear cells obtained from patients who received human leucocyte antigen (HLA)-matched allogeneic haematopoietic stem cell transplantation (HSCT). One of the allogeneic antigen-specific CD4+ CTL lines, CTL-A, generated from a patient with T cell acute lymphoblastic leukaemia, recognised HLA-DPB1*0501-positive Epstein-Barr virus-immortalised human B cell line (EBV-B cells), phytohaemagglutinin blasts and leukaemia cells, but not interferon-gamma (IFN-gamma) treated HLA-DPB1*0501 positive fibroblasts, indicating that this CD4+ T-cell line recognised a minor histocompatibility antigen (mHa) that is preferentially expressed in haematopoietic cells in an HLA-DPB1*0501-restricted manner. The other CD4+ CTL line, CTL-B, generated from a patient with chronic myeloid leukaemia, recognised mismatched HLA-DQB1*0303 on EBV-B cells and phytohaemagglutinin (PHA) blasts. Interestingly, this CTL line did not recognise IFN-gamma-treated recipient's skin fibroblasts, as HLA-DQ was merely upregulated even after IFN-gamma stimulation in non-haematopoietic cells including fibroblasts, endothelial cells and hepatocytes. These results suggest that these CD4 positive CTLs, specific for mismatch HLA-DQ and mHa that are preferentially expressed on haematopoietic cells, may play an important role in induction of selective graft-versus leukaemia effect without development of graft-versus-host disease after allogeneic HSCT. PMID- 16371021 TI - Prognostic value of inhibitory anti-ADAMTS13 antibodies in adult-acquired thrombotic thrombocytopenic purpura. AB - In order to assess the prognostic value of inhibitory anti-ADAMTS13 antibodies in thrombotic thrombocytopenic purpura (TTP), we performed a multicentre prospective study of 33 adult patients with idiopathic acquired TTP. Patients were treated with high-dose plasma infusion and therapeutic plasma exchange. Patients without (group 1, n = 12) and with (group 2, n = 21) detectable inhibitory anti-ADAMTS13 antibodies were compared for clinical presentation, treatment and outcome. Both groups were comparable for clinical presentation. All patients in group 1 achieved a sustained complete remission within a median of 7 d [95% confidence interval (CI), 4-18], which required a median plasma volume of 235 ml/kg (range, 131-1251). In group 2, 17 patients achieved a durable complete remission within a median of 23 d (95% CI, 11-32) (P = 0.001). Median plasma volume was 718 ml/kg (range, 219-3107) (P = 0.02). In group 2, there was a trend for more episodes of flare-up than in group 1 (13 vs. 3, respectively, P = 0.07). Four patients, all from group 2, died (P = not significant). The relapse rate was comparable between both groups. We suggest that TTP with detectable inhibitory anti-ADAMTS13 antibodies displays a worse prognosis, relative to a delayed platelet count recovery, a higher plasma volume requirement to achieve complete remission, and a trend for more frequent episodes of flare-up. PMID- 16371022 TI - Harmonisation of factor VIII:C assay results: study within the framework of the Dutch project 'Calibration 2000'. AB - In a Dutch project for harmonisation of factor VIII coagulant activity (FVIII:C) assays, the commutability of potential calibrators for FVIII:C was assessed by means of a 'twin-study design', which is in essence a multi-centre, split-patient sample, between-field-methods protocol. Commutability was defined as the degree to which a material yielded the same numerical relationships between results of measurements by a given set of measurement procedures as those between the expectations of the relationships for the same procedures applied to those types of material for which the procedures were intended. The study consisted of the simultaneous analysis of fresh frozen patient plasmas and three potential calibrators for FVIII:C by 16 Dutch laboratories forming eight couples. The state of-the-art intra-laboratory standard deviation was used to assess the commutability of the potential calibrators. One potential calibrator was used to harmonise FVIII:C assay results in a Dutch field study. The inter-laboratory coefficient of variation of two test samples could be reduced significantly, but no significant effect was observed with three other test samples. We recommend that at least three different sample dilutions be used in each FVIII:C assay, in agreement with previous recommendations. PMID- 16371023 TI - Identification of quantitative trait loci regulating haematopoietic parameters in B6AKRF2 mice. AB - The haematopoietic system is a complex organised tissue with a hierarchical structure. Identification of organisational pathways within the haematopoietic system is relevant for a better understanding of haematopoiesis in health and disease. We have analysed numerous haematopoietic parameters in two panels of a total of 157 genetically distinct B6AKRF2 mice, derived from an intercross between AKR and C57Bl/6 mice, strains known to differ in various stem cell traits. The major objective of our study was to assess the extent to which various haematopoietic parameters, such as stem cell numbers, progenitor cell cycling, progenitor cell mobilisation and neutrophil numbers in blood and bone marrow are coregulated. The genotypes of these mice were used to search for genetic loci that regulate these parameters. We found significant quantitative trait loci (QTL) associated with the number of stem cells (CAFC-35) in the bone marrow and the number of neutrophils in the blood. However, most haematopoietic parameters appeared to be controlled by non-heritable (epigenetic) factors, or by multiple QTLs. Our study reveals striking differences in structure of the haematopoietic hierarchy between individual mice. Surprisingly, stem and progenitor cell pool size and proliferation rate, as well as peripheral blood cell counts are all independently regulated. PMID- 16371024 TI - NKp46 and NKG2D receptor expression in NK cells with CD56dim and CD56bright phenotype: regulation by histamine and reactive oxygen species. AB - The cytotoxicity of natural killer (NK) cells is dependent on the interaction between target cell ligands and a series of stimulatory receptors on NK cells. Two of these triggering receptors, the NKp46 natural cytotoxicity receptor (NKp46) and the major histocompatibility complex (MHC) class I-interactive NKG2D receptor, are deficiently expressed by NK cells recovered from patients with acute myeloid leukaemia (AML), but little is known regarding the regulation of NKp46 and NKG2D expression. Here we report that mononuclear and polymorphonuclear phagocytes downregulate the cell surface density of NKp46 and NKG2D on NK cells with CD56(dim) phenotype in vitro by a mechanism that is dependent on the availability of phagocyte-derived reactive oxygen species (ROS). Histamine maintained NKp46 and NKG2D expression despite the presence of inhibitory phagocytes by targeting an H2 receptor on phagocytes. By contrast, NKp46 and NKG2D expression by the CD56(bright) subset of NK cells was resistant to inhibition by phagocytes. Our findings are suggestive of a novel mechanism of relevance to the regulation of NKp46/NKG2D receptor expression. Moreover, our findings suggest that the previously reported action of histamine on NK cell mediated killing of leukaemic cells may be related to the preservation of activatory NK-cell receptors. PMID- 16371025 TI - An exploratory study of physiological correlates of neurodevelopmental delay in infants with sickle cell anaemia. AB - This study aimed to investigate whether infants with sickle cell anaemia (SCA) are at risk of neurodevelopmental delay, and whether any delay is associated with SCA pathology. Twenty-eight infants (14 SCA; 14 age- and ethnic-similar controls) were assessed longitudinally with the Bayley Infant Neurodevelopmental Screener (BINS) at 3, 9 and 12 months. Transcranial Doppler (TCD) and pulse oximetry (SpO2) measures were recorded longitudinally in SCA infants, and a subgroup of controls. Haemoglobin values were obtained from SCA infants. At each age, SCA infants obtained BINS scores indicative of greater risk of neurodevelopmental delay compared with controls. The number of moderate-high BINS risk scores increased significantly between 3 and 9 months. At 9 months BINS raw scores correlated negatively with TCD velocity and positively with haemoglobin. This exploratory study suggests that SCA infants may be at greater risk of neurodevelopmental delay than previously considered, and may provide the impetus for further research into the very early precursors of cognitive impairment. PMID- 16371026 TI - Red blood cells, platelets and polymorphonuclear neutrophils of patients with sickle cell disease exhibit oxidative stress that can be ameliorated by antioxidants. AB - Sickle cell disease (SCD) is basically a red blood cell (RBC) disorder characterised by sickling and haemolysis, but platelets and polymorphonuclear neutrophils (PMN) are also involved. Oxidative damage may play a role in the pathogenesis of SCD. Using flow cytometry, we measured oxidative-state markers simultaneously in RBC, platelets and PMN obtained from 25 normal donors, nine homozygous (SS) patients and six SS/beta-thalassaemia patients. Reactive oxygen species (ROS) and reduced glutathione (GSH) were measured following staining of blood samples with fluorescence probes and gating on specific subpopulations based on size and granularity. Ten- to 30-fold higher ROS production and 20-50% lower GSH content were found in RBC, platelets and PMN from SCD patients versus those of their normal counterparts. This could in part account for the clinical manifestations, such as haemolysis, a hypercoagulable state, recurrent bacterial infections and vaso-occlusive incidences, in SCD. We further showed that exposure of SCD samples to antioxidants, such as N-acetyl-cysteine, vitamin C and vitamin E, decreased their oxidative stress. These results suggest that antioxidant treatment of patients with SCD could reduce oxidative damage to RBC, PMN and platelets, thereby alleviating symptoms associated with their pathology. The flow cytometry techniques presented herein could assist in monitoring the efficacy of such treatment. PMID- 16371027 TI - Bisphosphonate-associated osteonecrosis of the auditory canal. PMID- 16371028 TI - Pyoderma gangrenosum complicating pegylated granulocyte colony-stimulating factor in Hodgkin lymphoma. PMID- 16371029 TI - Novel observation of three FLT3 codons mutated in tandem in an elderly acute myeloid leukaemia patient. PMID- 16371033 TI - Myiasis in a blood donor: is it a reason for deferral? PMID- 16371034 TI - Bacterial detection and extended platelet storage: the next step forward. PMID- 16371035 TI - Beyond leukodepletion: removing infectious prions by filtration. PMID- 16371036 TI - Removal of exogenous (spiked) and endogenous prion infectivity from red cells with a new prototype of leukoreduction filter. AB - BACKGROUND: Two recent probable cases of transmission of a variant of human Creutzfeldt-Jakob disease (vCJD) through blood transfusion suggest that the disease can be transmitted through transfusion of blood components from presymptomatic blood donors. In the absence of a preclinical screening test, removal of the infectious agent by processing is the only means by which risk to recipients of blood from donors with inapparent vCJD infections can be eliminated. STUDY DESIGN AND METHODS: In the endogenous infectivity study, a pool of 500 mL of whole blood was collected into CP2D anticoagulant from 263K-strain scrapie-infected hamsters, processed into 300 mL of red cells (RBCs), and then passed through a prion removal filter. Pre- and postfiltration samples were tested for PrP(sc) by Western blot and for infectivity by inoculation of healthy hamsters. In the exogenous (spiking) infectivity study, 30 mL of 10 percent (wt/vol) scrapie-infected brain homogenates was added to 270 mL of human RBCs and then filtered. Levels of PrP(sc) and infectivity were determined by Western blot and bioassay. RESULTS: In the endogenous infectivity study, the prefiltered RBCs transmitted disease to 6 of 43 animals, whereas the postfiltered RBCs did not transmit disease to any of 35 animals, and a barely visible prefiltration PrP(sc) Western blot signal was reduced below the level of detection in the postfiltration sample. In the exogenous (spike) study, infectivity was reduced by 3.7 log LD50 per mL, from 9.2 to 5.5 log LD50 per mL. CONCLUSION: The new filter was effective in removing both infectivity and PrP(sc) from RBCs. The use of this type of filter should reduce the risk of vCJD transmission through blood transfusion. PMID- 16371037 TI - Detection of bacterial contamination in apheresis platelet products: American Red Cross experience, 2004. AB - BACKGROUND: Routine quality control (QC) testing for bacterial contamination in apheresis platelet (PLT) products was implemented in all 36 regional blood centers of the American Red Cross in March 2004. STUDY DESIGN AND METHODS: PLT samples were cultured under aerobic conditions until the end of the product shelf life or when a positive reaction was indicated. To confirm the initial positive reaction, a new sample was taken from the unit for reculturing. All positive culture bottles were referred for bacterial isolation and identification. Bacterial testing data along with apheresis PLT collection information were collected for analysis. Reports and investigations of potential septic reactions to apheresis PLTs were reviewed. RESULTS: In the first 10 months of bacterial testing, 226 of 350,658 collections tested initially positive. Sixty-eight were confirmed on resampling to be bacterially contaminated for an overall confirmed positive rate of 0.019 percent or 1 in 5157. Staphylococcus spp. (47.1%) and Streptococcus spp. (26.5%) were the most frequently isolated bacteria; Gram negative bacteria accounted for 17.6 percent of the confirmed-positive products. Of the 354 apheresis PLT products derived from all 226 initial test-positive cases, 38 (10.7%) were transfused by the time the initial positive reaction was indicated. None of these transfused products, however, had a confirmed-positive bacterial screen and no patient who had been transfused with an unconfirmed positive product had evidence of a septic transfusion reaction. Three high probability septic transfusion reactions to screened, negative components were identified. In all three cases, a coagulase-negative Staphylococcus was implicated. CONCLUSION: Our experience demonstrates that bacterial testing of apheresis PLT products as a QC measure was efficiently implemented throughout the American Red Cross system and that this new procedure has been effective in identifying and preventing the transfusion of many, although not all, bacterially contaminated PLT units. PMID- 16371038 TI - Relative sensitivities of licensed nucleic acid amplification tests for detection of viremia in early human immunodeficiency virus and hepatitis C virus infection. AB - BACKGROUND: Screening donors for human immunodeficiency virus (HIV) and hepatitis C virus (HCV) RNA is primarily performed on minipools (MPs) with one of two commercial nucleic acid amplification tests (NAT; Roche Molecular Systems; or Gen Probe/Chiron). We compared these assays with respect to detection of RNA in early HIV and HCV infection. STUDY DESIGN AND METHODS: Twelve HIV plasma donor panels (116 serial samples) and 12 HCV panels (180 serial samples) were selected to optimally represent early viremia. Initial testing was performed in singlicate or triplicate on separately coded aliquots, both neat and at dilutions corresponding to MP screening (1:16 for Gen-Probe; 1:24 for Roche); 20 additional replicates were performed when discordant results were observed. Odds ratios (ORs) comparing detection of RNA by different assays were derived with logistic regression models. Differences in window-period closure and yields of assays in MP or individual-donation (ID) format were estimated. RESULTS: Differences in detection rates between Roche and Gen-Probe NAT assays were small and only observed with samples with very-low-level viremia. ORs for detecting RNA by the Gen-Probe versus the Roche assay were significant for HIV if conducted on MPs (1.8; 95% confidence interval [CI], 1.3-2.5) but not neat (1.0; 95% CI, 0.72-1.4). Odds of detecting HCV RNA were higher if the Gen-Probe assay was conducted either neat (2.3; 95% CI, 1.6-3.2) or on MPs (4.0; 95% CI, 2.8-5.8). These differences translated to <1 day window-period closure and 100 x 10(6) NCs/mL) in most cases. The effect of NC concentrations at freezing in NC recovery after thawing and engraftment kinetics was analyzed and compared with a group of HPC-A cryopreserved at standard NC concentrations (n = 455, Protocol A). RESULTS: The mean (SD) NC concentration at freezing was 78 (28) x 10(6) per mL (median, 82 x 10(6)/mL; range, 12 x 10(6)-156 x 10(6)/mL) and 183 (108) x 10(6) per mL (median, 156 x 10(6)/mL; range, 16 x 10(6)-678 x 10(6)/mL), for HPC-A cryopreserved according to Protocols A and B, respectively. The NC viabilities of the test vials and HPC-A components after thawing were 88 percent versus 85 percent and 85 percent versus 82 percent, and the cloning efficiency was 49 percent versus 33 percent for Protocols A and B, respectively (p < 0.001). Significant differences were not observed in the recovery of NCs. Days to neutrophil and platelet engraftment were not different between patients transplanted in the standard- (n = 143) or high-cell-concentration group (n = 238). CONCLUSION: The cryopreservation of HPC-A at higher than standard NC concentrations has no adverse impact on hematopoietic reconstitution after transplantation. PMID- 16371045 TI - Delayed and acute hemolytic transfusion reactions resulting from red cell antibodies and red cell-reactive HLA antibodies. AB - BACKGROUND: It has been controversial whether HLA antibodies cause hemolytic transfusion reactions (HTR) or shortened red blood cell (RBC) survival. A patient is reported who had two episodes of HTR, the latter of which was likely due to RBC-reactive HLA antibodies. CASE REPORT: A 77-year-old woman, admitted for gastric varix rupture, had no RBC-irregular antibodies detected before transfusion. On Hospital Day 12, after transfusion of 2 units of RBCs and 2 units of fresh-frozen plasma, the first delayed hemolytic episode occurred and anti-E, anti-c, anti-Jk(a), and unidentified RBC-reactive antibodies were detected in a serum sample from Day 14. Two additional units of matched RBCs were transfused with a leukoreduction filter on Days 19 and 22. After 4 hours of starting a transfusion on Day 22, the patient had fever, and a second hemolytic episode was recorded. Multireactive HLA antibodies (reactive against 20 of 20 donor panel lymphocytes) were detected in serum samples from Day 15 to Day 21. These HLA antibodies reacted strongly with HLA-A2 and HLA-B7 antigens, corresponding to Bg(c) and Bg(a) antigens on RBCs, respectively. RBCs transfused on Day 22 were found to be HLA-A2 by genotyping. CONCLUSION: Strong HLA alloantibodies in this recipient appear to have caused a HTR. It is suggested that HLA antibodies be considered in patients with unexplained HTRs. PMID- 16371046 TI - Hyperhemolysis syndrome in anemia of chronic disease. AB - BACKGROUND: Occasional cases of delayed hemolytic transfusion reaction (DHTR) demonstrate severe and persistent hemolysis and are referred to as hyperhemolysis syndrome. This syndrome usually occurs in patients with sickle cell disease and possibly thalassemia who receive multiple transfusions. There are few such clinical reports in patients without hemoglobinopathies. CASE REPORT: A 67-year old woman with anemia and a history of four previous transfusions was admitted with shortness of breath and a hematocrit (Hct) level of 27 percent. The patient was group O with a negative antibody screen. She received 1 unit of electronically cross-matched red blood cells (RBCs) and was discharged. Thirteen days later she returned to hospital with weakness and a Hct level of 23 percent. The antibody screen now demonstrated anti-K alloantibody. The direct antigloblulin test (DAT) was positive with both anti-immunoglobulin G and anti complement (C3). Anti-K was recovered in the eluate. The previously transfused RBC unit was positive for presence of the K antigen. The patient's RBCs were negative for the presence of K antigen. Other laboratory data confirmed ongoing hemolysis, and a diagnosis of DHTR was made. She continued to display findings of active hemolysis for 9 more weeks requiring 19 units of RBCs. Thirty-four days after the original transfusion, her DAT remained positive and both the plasma sample and a RBC eluate demonstrated anti-K. CONCLUSION: The delayed hemolytic transfusion reaction with hyperhemolysis can occur among patients without hemoglobinopathies. PMID- 16371047 TI - Denaturing high-performance liquid chromatography-based genotyping and genetic variation of FUT2 in Sri Lanka. AB - BACKGROUND: Human ABO-secretor locus (FUT2) is highly polymorphic in many human populations. STUDY DESIGN AND METHODS: The applicability of denaturing high performance liquid chromatography (DHPLC) analysis was evaluated for genotyping the FUT2 in two Sri Lankan populations (Tamil and Sinhalese). RESULTS: Although DHPLC failed to detect one allele, 302C>T, of eight alleles, this method reduces the number of samples to be sequenced and can detect novel polymorphisms by comparing the elution profiles. CONCLUSION: These results suggest that DHPLC analysis is a useful high-throughput method for genotyping FUT2 and, further, that the genetic backgrounds of two Sri Lankan populations are quite similar, with little genetic flow from neighboring East and Southeast Asian populations to Sri Lanka. PMID- 16371048 TI - SCER and SCAN: two novel high-prevalence antigens in the Scianna blood group system. AB - BACKGROUND: More than 20 years ago, two probands were described whose red blood cells (RBCs) typed Sc:1,-2,3. Their serum samples contained alloantibodies reactive with all RBCs tested except those of the Sc:-1,-2,-3 phenotype. Cloning of the Scianna gene allowed us to determine the molecular bases of these samples. STUDY DESIGN AND METHODS: In a collaborative effort, the two probands' samples and also two Sc:-1,-2,-3 samples were obtained from frozen storage. All 11 SC (ERMAP) exons and their flanking regions were sequenced. RESULTS: The two probands with antibodies to Scianna-related antigens were homozygous, respectively, for an ERMAP(R81Q) allele caused by a G to A substitution at nucleotide 242 in the ERMAP gene and for an ERMAP(H26Y,G35S) allele, in which the G35S substitution was caused by a G to A substitution at nucleotide 103. Two patients with the Sc:-1,-2,-3 phenotype both carried ERMAP(R332X) alleles caused by a C to T substitution at nucleotide 994 that differed at one nucleotide position in the noncoding region of exon 11. In eight samples carrying orphan low prevalence antigens, no ERMAP variants were detected that could be implicated in Scianna antigen expression. CONCLUSION: SCER and SCAN expanded the Scianna blood group system to seven antigens, have been assigned the ISBT numbers 013.006 (Sc6) and 013.007 (Sc7), and were associated with ERMAP(R81Q) and ERMAP(G35S) proteins, respectively. ERMAP(R332X) is a second molecular basis for the Sc(null) phenotype. The eight low-prevalence antigens By, To(a), Pt(a), Re(a), Je(a), Li(a), SARA, and Sk(a) do not belong to the Scianna blood group system. PMID- 16371049 TI - Prospective epidemiologic study of the outcome and cost-effectiveness of antenatal screening to detect neonatal alloimmune thrombocytopenia due to anti HPA-1a. AB - BACKGROUND: To assess the value of antenatal screening to detect neonatal alloimmune thrombocytopenia (NAIT) due to anti-HPA-1a, a prospective study was carried out to quantify the potential clinical benefits and determine whether screening would be cost-effective. STUDY DESIGN AND METHODS: An observational prospective controlled study was carried out on 26,506 pregnant women over 2 years. HPA-1a phenotyping was performed in the first trimester and women confirmed HPA-1a-negative were tested for anti-HPA-1a during pregnancy, at delivery, and 10 to 14 days after birth. Babies of HPA-1a-negative women were tested at delivery for thrombocytopenia and examined for signs of bleeding. Economic evaluation was undertaken on the basis of the data collected during the study. RESULTS: Twenty-five of 318 women (7.9%) had anti-HPA-1a detected for the first time. Eight women (43 per 100,000) gave birth to babies with NAIT, and 5 (27 per 100,000) had severe thrombocytopenia. Three babies had mild signs of bleeding, and no cases of intracranial hemorrhage (ICH) or fetal loss were detected. It is estimated that it would cost 60,596 pounds (98,771 US dollars) to detect a case of severe NAIT, where anti-HPA-1a has been identified for the first time, and 1,151,323 pounds (1,876,656 US dollars) to prevent a case of ICH, assuming that detection allowed successful intervention. CONCLUSIONS: Our data suggest that severe HPA-1a NAIT is underdiagnosed in the absence of routine antenatal screening. Serious bleeding complications and ICH, however, occur less frequently in first cases of NAIT than suspected from the literature, and the costs of screening and possible intervention must be balanced against the procedural risks. PMID- 16371050 TI - The impact of e-mail in acquiring and retaining whole-blood donors: a comparative analysis of the Puget Sound Blood Center donor e-mail communication program. AB - BACKGROUND: With the emergence of e-mail as a common form of communication, it is important to understand the role and impact e-mail can have on acquiring and retaining whole-blood donors. STUDY DESIGN AND METHODS: Whole-blood donors who opted in to receive e-mail communications (n = 43,232) from the Puget Sound Blood Center (PSBC) and as a result participated in one or more of five e-mail marketing campaigns between July 2002 and March 2003 were studied. RESULTS: New donors with e-mail addresses grew by 74 percent over the measurement period compared with 2 percent growth of those without e-mail addresses. Nearly 15,000 prospective donors were reached as a result of donors passing along e-mails they received from PSBC to friends and family. Nearly 5 percent of PSBC e-mail campaign recipients registered to donate online. PSBC e-mail campaigns have outperformed e-mail marketing industry mean open rates by 44 percent and click through rates by 36 percent. CONCLUSION: E-mail is now a mainstream communication medium. It has proven to be an effective method of reaching new donors and generating new donation registrations. Donors respond more favorably to e-mail communications from PSBC compared with e-mail they receive from other organizations. Deeper understanding of how different segments of blood donors use e-mail may help blood centers adjust their donor acquisition and retention programs. New recruitment methods are needed to grow the overall donor base, and e-mail appears to have the potential to make a significant contribution to meeting this objective. PMID- 16371051 TI - Is an assay for simultaneous detection of hepatitis C virus core antigen and antibody a valuable alternative to nucleic acid testing? AB - BACKGROUND: A new enzyme immunoassay based on the simultaneous detection of nucleocapsid proteins of hepatitis C virus (HCV) and anti-HCV (Monolisa HCV antigen-antibody Ultra, Bio-Rad) was evaluated as an alternative to nucleic acid testing (NAT) for the diagnosis of HCV infection during the window period in blood donations. STUDY DESIGN AND METHODS: The study included 107 sequential samples from 10 HCV seroconversion commercial panels; 81 samples were in the preseroconversion phase, and 26 were collected after seroconversion. All samples were tested with HCV antigen-antibody assay and the two minipool (MP) NAT procedures that are routinely used in France (transcription-mediated amplification in pools of 8 and COBAS AmpliScreen HCV test [Roche Diagnostic] in pools of 24 donations). RESULTS: From the 44 samples collected during window period that were MP-NAT-positive, 31 (70.5%) were also positive with the Monolisa HCV antigen-antibody assay. The mean delay in detecting HCV infection between these two methods was 5.1 days (range, 0-24 days). The Monolisa HCV antigen antibody assay led to a reduction in the window period of 26.8 days (range, 0-72 days). All samples collected after seroconversion were detected with the HCV antigen-antibody assay. The specificity analyzed in 2503 consecutive blood donations was estimated at 99.88 percent. CONCLUSION: This new developed assay presents an improvement for the detection of HCV infection, especially in the early phase of infection when antibodies are undetectable. Although less sensitive than NAT, this assay could be a suitable solution for blood screening in developing countries where NAT (or HCV core antigen-specific assay) is not affordable or its implementation is not feasible. PMID- 16371052 TI - Prevalence of viral markers among first-time Arab blood donors in Kuwait. AB - BACKGROUND: The aim of this study was to assess the effect of blood donation modes on the prevalence of viral markers among Arab first-time blood donors in Kuwait. STUDY DESIGN AND METHODS: Donor ethnic background was classified as Kuwaiti nationals and non-Kuwaiti Arabs. A total of 26,874 donors were screened in 2002 for the following viral markers: hepatitis C virus antibody (anti-HCV), hepatitis B surface antigen (HBsAg), anti-hepatitis B core antigen (HBc), human immunodeficiency virus-1 and -2 antibody (anti-HIV-1 and -2), HIV p24, and human T lymphotropic virus-I and -II antibody (anti-HTLVI/II). All samples positive for the presence of anti-HBc were tested for anti-HBs. Among these donors, 12,798 were first-time donors of which 74 percent were replacement and 26 percent were volunteers. RESULTS: The prevalence of HCV among replacement donors was significantly higher than the volunteer group. The difference between the two modes of blood donations, however, was not significant for HBsAg. The prevalence of anti-HCV among Kuwaiti national and non-Kuwaiti Arab first-time donors was 0.8 and 5.4 percent, respectively, whereas the prevalence of HBsAg was 1.1 and 3.5 percent, respectively, with the difference being significant at a p level of <0.0001. The difference observed for prevalence of anti-HBc among Kuwaiti national and non-Kuwaiti Arab donors (17 and 33.3%, respectively) was significant (p < 0.0001). Among first-time donors, 13.7 percent were positive for the presence of anti-HBs, indicating that 13.7 percent of the total Arab donor population might have had a previous infection and possible immunity to hepatitis B virus (HBV). CONCLUSION: A high prevalence of HBV and HCV was found among non Kuwaiti Arab donors. The prevalence of anti-HCV was only significantly higher among replacement versus volunteer first-time donors. Therefore, there is a need to develop a strategic plan that incorporates the diverse background of the blood donors living in Kuwait. PMID- 16371055 TI - Platelet transfusion in thrombotic thrombocytopenic purpura: between Scylla and Charybdis. PMID- 16371056 TI - ABO-mismatched marrow transplantation: removal of mismatched donor red blood cells by "washing" with recipient's matched red blood cells in a closed system. PMID- 16371058 TI - Survival of fetal red blood cells. PMID- 16371060 TI - Anomalous response to potassium in vascular smooth muscle cells of human saphenous vein. AB - 1.--We have examined the relationship between the resting membrane potential (E(m)) and the concentration of the external ions, K(+), Cl(-) and Ca(2+), as well as the effects of K(+) on active force generation in human saphenous veins. 2.--As measured with sharp glass microelectrodes, the E(m) of vascular muscle cells was -76.0 +/- 7.0 mV (mean +/- SD; n = 328). Raising the concentration of external potassium ([K(+)](e)) from 4.2 to 20, 40, 80, 120 or 150 mm produced an incremental depolarization, revealing a maximal slope factor of 15 mV per 10-fold increase. 3.--Oubain (1.0 microm) did not have any effect on E(m) (-79.0 +/- 8.0 mV; n = 80). Replacement of external Cl(-) with propionate resulted in significant (P < 0.05) depolarization (E(m): -65.5 +/- 7.5 mV; n = 40). In Cl(-) free buffer containing 80 mm K(+), E(m) depolarized to -52.0 +/- 6.7 mV (n = 45) compared with -64.7 +/- 6.5 mV (n = 55) (P < 0.05) measured in buffer containing 80 mm [K(+)](e) and Cl(-) 138.7 mm. Removal of Ca(2+) did not significantly modify the depolarizing response to K(+) 80 mm: E(m), -68.2 +/- 4.9 mV (n = 42) vs.-64.7 +/- 6.5 mV (n = 55) in the presence of Ca(2+). 4.--Despite their small size, changes in E(m) correlated closely with force generation in buffer containing high K(+), approximately 3.62 mN force being generated per mV of change in E(m). 5.--These data demonstrate that, in human saphenous smooth muscle cells, (i) the magnitude of depolarization induced by raising [K(+)](e) deviates considerably from the theoretical values predicted by the Goldman-Hodgkin-Katz equations, (ii) Cl(-) appears to contribute to the maintenance of E(m), and (iii) electromechanical coupling has a low threshold. PMID- 16371061 TI - Relaxant responses to calcium channel antagonists and potassium channel opener in human saphenous vein. AB - 1.-- As shown in a parallel study the magnitude of depolarization induced in human saphenous vein by raising external potassium ([K(+)](e)) falls markedly below the theoretical values predicted by the Goldman-Hodgkin-Katz equations. This anomaly prompted us to re-examine the relaxant actions of L-type (nifedipine) and T-type (mibefradil) Ca(2+) channel antagonists, and relaxant and electrophysiological effects of the K(+) channel opener, pinacidil, on saphenous veins contracted by the elevation of [K(+)](e). 2.-- Nifedipine produced concentration-dependent relaxations in tissues contracted at various high [K(+)](e). In tissues contracted with 20 mm [K(+)](e), the pIC(50) for nifedipine was significantly (8.20 +/- 0.05; n = 6; mean +/- SEM; P < 0.05) greater than in tissues contracted with > or =40 mm [K(+)](e). 3.-- Tissues contracted with 20 mm [K(+)](e) also relaxed in response to mibefradil (pIC(50) = 6.1 +/- 0.14) and pinacidil (pIC(50) = 6.45 +/- 0.08), the latter being almost completely reversed (93.4 +/- 9.9%) by addition of glibenclamide (10 microm). 4.-- The resting E(m) of smooth muscle cells of saphenous vein was -77.0 +/- 0.7 mV (n = 52), and 20 mm [K(+)](e) produced a modest but significant depolarization to -73.0 +/- 0.7 mV (n = 52). Incubation with pinacidil plus 20 mm [K(+)](e) resulted in a significant hyperpolarization of the E(m) to -82 +/- 0.6 mV (n = 52). 5.-- N(omega)-nitro-L arginine methyl ester did not impede the relaxant responses of nifedipine, mibefradil or pinacidil. 6.-- In conclusion, the relaxant effects of nifedipine and pinacidil (i) occurred at an E(m) distinctly below the presumed threshold for the opening of the classic (Ca(V)1.3alpha(1)) L-type Ca(2+) channels, and (ii) did not depend on generation of nitric oxide. PMID- 16371062 TI - Oral treatment and in vitro incubation with fructose modify vascular prostanoid production in the rat. AB - 1.-- In the rat, a fructose-enriched diet induces hyperglycaemia, hypertriglyceridaemia, insulin resistance and hypertension; a model which resembles the human metabolic syndrome. 2.-- Prostanoids, metabolites of arachidonic acid, include vasoactive substances synthesized and released from the vascular wall that have been implicated in the increase of peripheral resistance, one of the mechanisms involved in the fructose-induced hypertension. 3.-- The aim of the present study was to: (i) analyse the effects of the in vitro incubation with fructose on the production and release of prostanoids in rat thoracic aorta and in rat mesenteric bed and (ii) compare the effects of incubation with those of the in vivo acute and chronic treatment of rats with fructose and with the combination of both in vivo and in vitro procedures. 4.-- Blood pressure, glycaemia and triglyceridaemia were significantly elevated in both 4- and 22-week fructose-treated groups. Meanwhile, body and heart weight as well as insulinaemia were similar between experimental animals and controls. 5.-- In aortae, 4 weeks of Fructose treatment did not modify the prostanoid pattern release, but in vitro incubation decreased prostacyclin (PGI(2)) production. However, after 22 weeks, fructose treatment and incubation exerted the same effect. 6.-- In mesenteric bed, after 4 weeks, the incubation and the combination of both procedures reduced the release of the vasodilators PGI(2) and PGE(2), while fructose treatment only diminished the PGE(2) release. On the contrary, the production of the vasoconstrictor thromboxane A(2) (TXA(2)) was enhanced by incubation and both the procedures. After 22 weeks, fructose treatment increased PGI(2) release, while it was reduced by incubation. The combination of both did not modify this peripheral resistance when compared with controls. Finally, incubation of tissues from treated rats increased the release of the vasoconstrictors, PGF(2alpha) and TXA(2). 7.-- In conclusion, the mesenteric bed, a resistance vascular bed, seems to be more sensitive than the aorta, a conductance vessel, to the effects of fructose on prostanoid production. This difference could be related to a more relevant role of resistance vessels in the regulation of peripheral resistance and consequently of blood pressure. The observed effects should contribute to a shift in the balance of the release of prostanoid in favour of vasoconstrictor metabolites. This phenomenon could be related to an increase in the peripheral resistance and the mild hypertension observed in the fructose-treated rats. PMID- 16371063 TI - Captopril therapy decreases both expression and function of alpha-adrenoceptors in pre- hypertensive rat aorta. AB - 1.-- The effects of captopril on alpha(1)-adrenoceptor mRNA and protein and phenylephrine-induced contraction was assessed in aorta of pre-hypertensive spontaneously hypertensive rats. 2.-- Four-week-old SHR and WKY rats were treated with captopril [an angiotensin-converting enzyme (ACE) inhibitor] 3 mg kg(-1) day(-1) for 1 week. 3.-- pA(2) values for BMY 7378, an alpha(1D)-adrenoceptor antagonist, were 8.63-9.20 among the different groups. Schild slopes were close to unity suggesting that contraction was produced primarily by alpha(1D) adrenoceptor stimulation and was not changed with therapy. 4.-- Alpha(1D) adrenoceptor mRNA and protein values were higher in pre-hypertensive SHR than in WKY, whereas alpha(1A)-adrenoceptor mRNA was higher in WKY and alpha(1B) adrenoceptors were similar in both strains, and protein was not significantly different for alpha(1A)- and alpha(1B)-subtypes. 5.-- Captopril decreased maximal contraction in SHR, without having effect in WKY rats, while alpha(1D) adrenoceptor mRNA was decreased in both rat strains but alpha(1D)-adrenoceptor protein was significantly decreased only in SHR, and increased alpha(1A)-mRNA in SHR, no effect of captopril treatment was observed on alpha(1B)-adrenoceptor mRNA and protein nor on alpha(1A)-adrenoceptor protein. 6.-- These data suggest that ACE inhibition by captopril influences both expression and function of alpha(1D) adrenoceptors in aorta of pre-hypertensive rats, probably avoiding alpha(1D) subtype expression by blockade of angiotensin II synthesis. PMID- 16371066 TI - Children's exposure to secondhand tobacco smoke. PMID- 16371067 TI - The influence of ethnicity and generational status on parent and family relations among Chinese and Filipino adolescents. AB - OBJECTIVE: The purpose of this study was to explore the influence of ethnic and generational differences on perceptions about parent and family relationships in a nationally representative sample of Chinese and Filipino adolescents living in the United States. DESIGN: This was a cross-sectional descriptive study, utilizing data from the National Longitudinal Study on Adolescent Health. SAMPLE: The sample was weighted to produce a nationally representative sample, based on 216 self-identified Chinese and 387 self-identified Filipino adolescents. RESULTS: Results indicated that generational status plays a role in predicting how adolescents perceive the parent-adolescent relationship. CONCLUSIONS: Findings provide a basis by which community health nurses can begin to understand the uniqueness of Asian subgroups, as well as generations of immigrant youth. PMID- 16371068 TI - Incestuous experience among Korean adolescents: prevalence, family problems, perceived family dynamics, and psychological characteristics. AB - OBJECTIVES: The present study aimed to identify the prevalence of incest among Korean adolescents and to determine the family problems, perceived family dynamics, and psychological consequences associated with incest in South Korea. DESIGN: A cross-sectional study was performed, using an anonymous, self-reporting questionnaire. Data were analyzed by percentages, Chi-square, and t-test statistical analyses, using SAS software program. SAMPLE: A total of 1,672 adolescents (1,053 student adolescents and 619 delinquent adolescents) were selected using proportional stratified random sampling method in this study. RESULTS: The results showed a 3.7% prevalence of incest in the tested Korean population. Families in which incest occurred were characterized by higher levels of problems, such as psychotic disorders, depression, criminal acts, and alcoholism among family members. Adolescent incest victims showed significantly more dysfunctional and unhealthy in terms of family dynamics and expressed significantly higher maladaptive and problematic psychological patterns than nonvictimized adolescents. CONCLUSIONS: The present findings identified some of the family problems and dysfunctional family dynamics may associate with intrafamilial child sexual abuse in Korea. Therefore, when an allegation of intrafamilial sexual abuse is made, health professionals should carry out a comprehensive assessment of their family dynamics and an evaluation of the impact of the abuse on the child and family. PMID- 16371069 TI - Health-risk behaviors in a sample of first-time pregnant adolescents. AB - OBJECTIVE: The purpose of this study was to assess the frequency of prenatal health-risk behaviors (substance use, sexual risk taking, and prenatal class attendance) among a nonrandom sample of first-time pregnant adolescents. DESIGN: The design is descriptive. SAMPLE: 145 ethnically diverse first-time pregnant adolescents aged 15-18 years. MEASUREMENT: Health behavior questions modified from the Center for Disease Control's Youth Risk Behavior Surveillance System. RESULTS: The health-risk behavior most modified during pregnancy was alcohol use (64/145 drank but quit and 1/145 did not quit). Of the 52/145 who used street drugs, nine continued despite pregnancy. Of the 75/145 who smoked early in pregnancy, 39 continued. The majority did not use a condom at last sexual intercourse. Approximately half attended a prenatal class and half attended a teen parenting class. CONCLUSION: Health-risk behaviors captured by birth certificate data are thought to be underreported for all age groups, and the prevalence of health-risk behaviors in this sample of pregnant teens was often greater than the most recent national trend data available. The magnitude of the effects of health-risk behaviors on pregnancy outcomes necessitates improved data gathering to enhance planning and evaluation of research and interventions at community, system, and individual/family levels. PMID- 16371070 TI - Predicting smoking status in a college-age population. AB - OBJECTIVE: The purpose of this study was to determine the factors that predict smoking group membership in a college-age population. DESIGN: A nonexperimental cross-sectional design was used to examine factors that distinguished among four groups: never established (n = 591), former (n = 41), nondaily (n = 80), and daily (n = 76) cigarette smokers. SAMPLE: A pencil and paper questionnaire was returned by 788 randomly selected college students, 18-24 years of age. RESULTS: Significant differences were found among the groups on their mothers', fathers', siblings', and peers' smoking behaviors and attitudes toward smoking. The nondaily and daily smoking groups reported a greater number of risk behaviors than the never established and former smoking groups. The two groups of current smokers also were more likely to report an increased level of depressive symptoms compared with the nonsmoking groups. The number of high school and college friends who smoke and self-reported high-risk alcohol and drug behaviors predicted smoking group membership in a polytomous logistic regression. CONCLUSIONS: Strategies to alter the social environment and decrease the exposure of nonsmoking students to others who model smoking behaviors may help decrease smoking initiation and prevent relapse on college campuses. PMID- 16371071 TI - Perceptions of health and well-being among women in a work-based welfare program. AB - Women who are single parents, poor, and employed in low-paying jobs have little choice about being dependent on public assistance programs to meet basic personal and family needs. OBJECTIVE: To explore women's perceptions of their health and well-being while enrolled in a work-based welfare program. This is the second in a series of articles about Temporary Assistance for Needy Families. DESIGN AND SAMPLE: Qualitative interviews and quantitative survey methods were used to explore perceptions of health and well-being in a purposive sample of mothers (n = 34) enrolled in a work-based welfare program in a city in the U.S. Midwest. METHODS: Instruments used were a semistructured interview guide, a demographic data form, and the General Well-Being Schedule (i.e., a survey tool developed for the U.S. Health and Nutrition Examination Survey). Data collection was completed in June 2000. The data were triangulated; using both quantitative and qualitative data added to the in-depth understanding of the subjects. RESULTS: Distress levels reported by participants were significantly higher than in the general U.S. population. CONCLUSIONS: Results of this study suggest that current policies do not effectively support health and well-being of single mothers enrolled in work-based welfare programs. PMID- 16371072 TI - Self-rated health and risk factors for metabolic syndrome among middle-aged men. AB - OBJECTIVES: To examine lifestyle and clinical risk factors for metabolic syndrome (MBO) and compare their significance between levels of self-rated health among middle-aged men. DESIGN: A cross-sectional baseline study. SAMPLE: 273 men, aged 40, living in Helsinki, Finland. METHODS: Postal questionnaires and health examinations by public health nurses were used in data collection. Statistical differences between groups of self-rated health and risk factors were analyzed by chi-square tests. RESULTS: Of all the respondents, 55% rated their health as good and 45% as average. Two thirds were overweight or obese, and 35% had waist-hip ratio more than 100 cm. Approximately 43% had diastolic blood pressure greater than 90 mmHg. Over half of the men smoked daily, and 28% used alcohol excessively. CONCLUSIONS: The men in this sample were found to be at high risk of developing MBO. The results underscore the importance of understanding the contradiction that exists between subjective and objective health ratings. Public health nurses are in a key position to educate men on how to use simple measurements to objectively assess their risk factors and, thus, potentially reduce their risk of developing diabetes, heart attack, or stroke. PMID- 16371073 TI - Re-evaluating current public health policy: alternative public health nursing approaches to sexually transmitted infection testing for teens and males who have sex with males. AB - OBJECTIVE: Due to a recent increase in the rates of reportable sexually transmitted infections (STI) across Canada, and the movement of public health nursing initiatives to a health promotion/illness prevention model, this paper proposes alternative strategies to current public health initiatives to encourage individuals who are labelled as at-risk by public health discourses, such as teens and males who have sex with males (MSM), to undertake preventative screening. THEORETICAL DESIGN: To undertake this task, we approached Health Canada's (2002) determinants of health utilizing Lupton's (1995) post structuralist work on public health to suggest that for some members of these groups, health is not an imperative. CONCLUSIONS: As such, we propose that increasing anonymous testing and its advertisement would provide one means of providing population-sensitive care and could thus increase the screening rates of individuals within groups who reject the public health "healthy" disquisition. PMID- 16371074 TI - Feeding better food habits in mid-20th-century America. AB - Public health nursing work has always involved education about nutrition and food habits. Nurses serve as interpreters of scientific and medical knowledge and as agents of behavioral change among the individuals and groups for whom they provide care. For public health nurses in mid-20th-century America, this meant direct involvement in many aspects of family life at home. Meal preparation and the eating patterns of family members were two areas into which most public health nurses had access. This brief history provides an introduction to some of the issues confronting American public health nurses at mid-century. Examining the content of articles related to nutrition in families around mid-century yields evidence about the role of the nurse, the state of the art in nutritional knowledge, and opinions and prejudices related to eating habits prevalent at the time. These glimpses into concerns expressed in the original Public Health Nursing journal may enhance understanding of the role of public health nurses in preventing diet-related disorders and form the basis for additional historical research. PMID- 16371075 TI - Comment: preventing child obesity -- an art and a science. PMID- 16371076 TI - Reducing obesity and related chronic disease risk in children and youth: a synthesis of evidence with 'best practice' recommendations. AB - Childhood obesity is a global epidemic and rising trends in overweight and obesity are apparent in both developed and developing countries. Available estimates for the period between the 1980s and 1990s show the prevalence of overweight and obesity in children increased by a magnitude of two to five times in developed countries (e.g. from 11% to over 30% in boys in Canada), and up to almost four times in developing countries (e.g. from 4% to 14% in Brazil). The goal of this synthesis research study was to develop best practice recommendations based on a systematic approach to finding, selecting and critically appraising programmes addressing prevention and treatment of childhood obesity and related risk of chronic diseases. An international panel of experts in areas of relevance to obesity provided guidance for the study. This synthesis research encompassed a comprehensive search of medical/academic and grey literature and the Internet covering the years 1982-2003. The appraisal approach developed to identify best practice was unique, in that it considered not only methodological rigour, but also population health, immigrant health and programme development/evaluation perspectives in the assessment. Scores were generated based on pre-determined criteria with programmes scoring in the top tertile of the scoring range in any one of the four appraisal categories included for further examination. The synthesis process included identification of gaps and an analysis and summary of programme development and programme effectiveness to enable conclusions to be drawn and recommendations to be made. The results from the library database searches (13,158 hits), the Internet search and key informant surveys were reduced to a review of 982 reports of which 500 were selected for critical appraisal. In total 158 articles, representing 147 programmes, were included for further analysis. The majority of reports were included based on high appraisal scores in programme development and evaluation with limited numbers eligible based on scores in other categories of appraisal. While no single programme emerged as a model of best practice, synthesis of included programmes provided rich information on elements that represent innovative rather than best practice under particular circumstances that are dynamic (changing according to population subgroups, age, ethnicity, setting, leadership, etc.). Thus the findings of this synthesis review identifies areas for action, opportunities for programme development and research priorities to inform the development of best practice recommendations that will reduce obesity and chronic disease risk in children and youth. A lack of programming to address the particular needs of subgroups of children and youth emerged in this review. Although immigrants new to developed countries may be more vulnerable to the obesogenic environment, no programmes were identified that specifically targeted their potentially specialized needs (e.g. different food supply in a new country). Children 0-6 years of age and males represented other population subgroups where obesity prevention programmes and evidence of effectiveness were limited. These gaps are of concern because (i) the pre-school years may be a critical period for obesity prevention as indicated by the association of the adiposity rebound and obesity in later years; and (ii) although the growing prevalence of obesity affects males and females equally; males may be more vulnerable to associated health risks such as cardiovascular disease. Other gaps in knowledge identified during synthesis include a limited number of interventions in home and community settings and a lack of upstream population based interventions. The shortage of programmes in community and home settings limits our understanding of the effectiveness of interventions in these environments, while the lack of upstream investment indicates an opportunity to develop more upstream and population-focused interventions to balance and extend the current emphasis on individual-based programmes. The evidence reviewed indicates that current programmes lead to short-term improvements in outcomes relating to obesity and chronic disease prevention with no adverse effects noted. This supports the continuation and further development of programmes currently directed at children and youth, as further evidence for best practice accumulates. In this synthesis, schools were found to be a critical setting for programming where health status indicators, such as body composition, chronic disease risk factors and fitness, can all be positively impacted. Engagement in physical activity emerged as a critical intervention in obesity prevention and reduction programmes. While many programmes in the review had the potential to integrate chronic disease prevention, few did; therefore efforts could be directed towards better integration of chronic disease prevention programmes to minimize duplication and optimize resources. Programmes require sustained long term resources to facilitate comprehensive evaluation that will ascertain if long term impact such as sustained normal weight is maintained. Furthermore, involving stakeholders in programme design, implementation and evaluation could be crucial to the success of interventions, helping to ensure that needs are met. A number of methodological issues related to the assessment of obesity intervention and prevention programmes were identified and offer insight into how research protocols can be enhanced to strengthen evidence for obesity interventions. Further research is required to understand the merits of the various forms in which interventions (singly and in combination) are delivered and in which circumstances they are effective. There is a critical need for the development of consistent indicators to ensure that comparisons of programme outcomes can be made to better inform best practice. PMID- 16371077 TI - New motilin agonists: a long and winding road. PMID- 16371078 TI - Functional gastrointestinal disorders and mast cells: implications for therapy. AB - The pathophysiology of functional gastrointestinal disorders is poorly understood. Accepted common mechanisms include psychosocial factors, abnormal gastrointestinal motility and disturbed visceral sensory perception, but the underlying causes remain unclear. Mast cells (MCs) are immunocytes widely distributed throughout the gastrointestinal tract. Several stimuli (e.g. allergens, neuropeptides and stress) lead to MC activation with consequent mediator release (e.g. histamine, tryptase and prostanoids). The MC mediators interact with nerves supplying the gut leading to altered gut physiology and increased sensory perception. The intestinal mucosa of irritable bowel syndrome patients contains on average an increased number of MCs. These cells release an increased amount of mediators in close vicinity to mucosal innervation. The MC activation and their close proximity to nerve fibres is correlated with the severity of perceived abdominal painful sensations. These data provide a strong basis for considering MCs as important participants in visceral hypersensitivity and pain perception in irritable bowel syndrome. Inhibition of MC function may ameliorate irritable bowel symptoms. Novel drugs with an increased potential in the control of MC function (e.g., anti-IgE antibodies, the intracellular protein tyrosine kinase inhibitor Syk) and mediator release (e.g., second generation antihistamines, proteinase-activated receptor antagonists) may be useful pharmacological tools for these common disorders. PMID- 16371079 TI - Symptom responses, long-term outcomes and adverse events beyond 3 years of high frequency gastric electrical stimulation for gastroparesis. AB - The aims were to determine symptom responses and long-term outcomes in gastroparetic patients receiving gastric electrical stimulation (GES) therapy beyond 3 years by presenting per protocol analysis and intention-to-treat (ITT) analysis. Data collected at baseline, 1 year and beyond 3 years in 55 patients included total symptom scores (TSS), nutritional status, weight, hospitalizations, the use of prokinetic and/or antiemetic medications, HbA1c in diabetics and adverse events. Of the 55 patients, 10 died of non-pacemaker related complications, six had the devices removed and two could not be reached. The remaining 37 patients had the device activated for a mean of 45 months. Both per protocol and ITT analysis demonstrated that TSS, hospitalization days and the use of medications were all significantly reduced at 1 year and were sustained beyond 3 years. Average TSS decreased by 62.5% for the 37 patients completing 3 years of GES. At implantation, 15/37 patients required nutritional support and only five continued beyond 3 years. Mean HbA1c level in diabetics was significantly reduced from 9.5 to 7.9% at 3 years. We conclude that a significant improvement in symptoms and all measures of clinical outcome can be maintained for greater than 3 years with GES in patients with refractory gastroparesis. PMID- 16371080 TI - Effect of atilmotin on gastrointestinal transit in healthy subjects: a randomized, placebo-controlled study. AB - We studied effects of i.v. atilmotin (BAX-ACC-1638, a novel motilin agonist, circulating t(1/2) < 10 min) on gastrointestinal transit in humans using a randomized, parallel-group, dose-response double-blind study of i.v. atilmotin, 6, 30, 60 microg or vehicle (placebo) given 2 min after standardized breakfast, lunch and dinner. The breakfast meal contained (99m)Tc-eggs and (111)In-milk. Full gastrointestinal transit was measured by scintigraphy. Primary endpoints were % gastric emptying (GE) at 30 min, GE t(1/2), colonic filling (CF) at 6 h, and geometric centre of colonic transit at 24 h. Analysis included adjustment for age, gender and body mass index, with Bonferroni correction applied for multiple comparisons. A significant treatment effect of atilmotin was detected for GE (%) at 30 min for solids and liquids (P < 0.01 for both). There were no significant effects on CF or CT and no significant adverse clinical events. Thus, atilmotin accelerates GE of solids and liquids in healthy humans. These data suggest that, at the doses tested, atilmotin should be considered for treatment of stomach motility disorders. PMID- 16371081 TI - Removal of tonic nitrergic inhibition is a potent stimulus for human proximal colonic propagating sequences. AB - Propagating sequences (PS) are important in colonic propulsion and defecation, yet the triggers of these motor patterns are not understood. Nonadrenergic noncholinergic neurones are believed to modulate smooth muscle in the gastrointestinal tract via the ubiquitous inhibitory neurotransmitter nitric oxide (NO). In the mouse colon periods of quiescence correlate with an increase in the release of NO. We investigated the colonic response to NO synthase inhibition in the conscious human subject. Intravenous infusion of saline or N(G) monomethyl-L-arginine (L-NMMA; 3 or 6 mg kg(-1) h(-1)) occurred in random order in six healthy volunteers in whom a 5 m long nasocolonic manometry catheter was positioned such that 16 recording sites, at 7.5-cm intervals, spanned the terminal ileum and colon. L-NMMA infusion at 3 mg kg(-1) h(-1), but not 6 mg kg( 1) h(-1) significantly (P = 0.02) increased proximal colonic PS frequency (2.0 +/ 1.9 vs 11.7 +/- 7.0 PS h(-1)) and non-propagating motor activity (5,296 +/- 2,750 vs 6,362 +/- 1,275 mmHg s). We conclude that blockade of NO synthesis has a stimulatory effect on the frequency of proximal colonic PS. This suggests removal of tonic nitrergic inhibition of the colon might be a physiological stimulus for propagating activity. PMID- 16371082 TI - Low-dose lactulose produces a tonic contraction in the human colon. AB - Lactulose (10-20 g day(-1)) is used to treat constipation. At this therapeutic dose, its effects on colonic motility remain unknown. Twenty-two healthy subjects swallowed a probe with an infusion catheter, six perfused catheters and a balloon connected to a barostat. Colonic phasic and tonic motor activity was recorded in fasting state. In group 1, four volunteers ingested 15 g lactulose and motility was recorded for 5 h after entry of lactulose into the caecum; in group 2, motility was recorded during (3 h) and 2 h after intracolonic infusion of isoosmotic and isovolumetric solutions containing sodium chloride alone (n = 9) or with 15 g lactulose (n = 9). In a last group of volunteers, isotopic colonic transit after ingestion of lactulose (10 g,n = 9) was assessed and compared with a control group (n = 17). Ingestion or intracolonic infusion of 15 g lactulose significantly decreased barostat bag volume (maximal decrease: 45 +/- 12% and 35 +/- 9% of basal value respectively). Phasic contractions remained unchanged. Tonic and phasic motility was unchanged by the isotonic and isovolumetric infusion of saline. Ingestion of lactulose significantly accelerated isotopic colonic transit time compared with the control group. We conclude that in healthy humans, 10-15 g ingestion or intracaecal infusion of lactulose produces a prolonged tonic contraction that may be involved in the laxative effect of lactulose. PMID- 16371083 TI - Motility disorder in experimentally obstructed intestine: relationship between muscularis inflammation and disruption of the ICC network. AB - We designed a model of small intestinal obstruction in rats to investigate changes in intestinal contractility associated with the immunologically activated components in the tunica muscularis. Although histochemical study did not reveal any typical inflammatory signs such as leucocyte infiltration in the distended intestinal regions of model rats 2-3 weeks after surgical induction of intestinal obstruction, the number of ED2-positive macrophages appeared to be increased in the tunica muscularis. Expression of tumour necrosis factor (TNF)-alpha mRNA was also significantly increased, and the level of CD14 was also increased significantly in the tunica muscularis. Functional studies of distended intestinal muscle segments showed a marked decrease in absolute force stimulated by a cholinergic agent. In addition, the number of spontaneous rhythmic contractions was also reduced in the distended intestinal regions of the obstructed intestine, and this decrease was associated with a reduction in the number of interstitial cells of Cajal (ICC), as revealed by Kit-like immunoreactivity. These results suggest that, under the pro-inflammatory conditions of the tunica muscularis associated with intestinal obstruction, the release of bioactive substances, possibly from activated resident macrophages, may affect smooth muscle contractility. Furthermore, under these conditions, both the number and the function of neighbouring ICC may also be affected. PMID- 16371084 TI - Neural mechanisms involved in the inhibition of intestinal motility induced by intestinal electrical stimulation in conscious dogs. AB - The effects and mechanisms of intestinal electrical stimulation (IES) with long pulses on intestinal motility were investigated in conscious dogs. Eighteen dogs were equipped with serosal electrodes and an intestinal cannula in the small bowel. The first experiment was designed to study the effect of one-channel IES on intestinal motility and the extent of this effect. The second experiment was conducted to study the effect of IES on intestinal motility and the involvement of neural pathway. The IES with long pulses significantly inhibited intestinal motility. Intestinal motility of the entire measured segment (40-220 cm distal to the stimulation electrodes) was inhibited by 60-74% with the single-channel IES with long pulses. Hexamethonium, guanethidine, phentolamine, propranolol partially, but not N(omega)-nitro-L-arginine (L-NNA), ondansetron and naloxone prevented the inhibitory effect of IES on intestinal motility. We conclude that single-channel IES inhibits intestinal motility within a distance of at least 2 m. This inhibitory effect induced by IES with long pulses is mediated via sympathetic but not nitrergic, serotoninergic 5-HT(3) and opiate pathway. PMID- 16371085 TI - The hallucinogenic herb Salvia divinorum and its active ingredient salvinorin A inhibit enteric cholinergic transmission in the guinea-pig ileum. AB - Salvia divinorum is a widespread hallucinogenic herb traditionally employed for divination, as well as a medicament for several disorders including disturbances of gastrointestinal motility. In the present study we evaluated the effect of a standardized extract from the leaves of S. divinorum (SDE) on enteric cholinergic transmission in the guinea-pig ileum. SDE reduced electrically evoked contractions without modifying the contractions elicited by exogenous acetylcholine, thus suggesting a prejunctional site of action. The inhibitory effect of SDE on twitch response was abolished by the opioid receptor antagonist naloxone and by the kappa-opioid antagonist nor-binaltorphimine, but not by naltrindole (a delta-opioid receptor antagonist), CTOP (a mu-opioid receptor antagonist), thioperamide (a H(3) receptor antagonist), yohimbine (an alpha(2) receptor antagonist), methysergide (a 5-hydroxytryptamine receptor antagonist), N(G)-nitro-L-arginine methyl ester (an inhibitor of NO synthase) or apamin (a blocker of Ca(2+)-activated K(+) channels). Salvinorin A, the main active ingredient of S. divinorum, inhibited in a nor-binaltorphimine- and naloxone sensitive manner electrically induced contractions. It is concluded that SDE depressed enteric cholinergic transmission likely through activation of kappa opioid receptors and this may provide the pharmacological basis underlying its traditional antidiarrhoeal use. Salvinorin A might be the chemical ingredient responsible for this activity. PMID- 16371086 TI - Activation of peripheral 5-HT receptors attenuates colonic sensitivity to intraluminal distension. AB - Tegaserod is a 5-HT(4) receptor partial agonist approved for the treatment of irritable bowel syndrome in women with constipation and in both men and women with chronic constipation. The efficacy of tegaserod is based on the importance of 5-HT(4) receptors regulating intestinal peristalsis and secretion, and possibly visceral sensory pathways. Our aim was to investigate the effect of tegaserod on colorectal sensitivity using models of normal and exaggerated responsiveness to colorectal distension (CRD). The visceromotor responses (VMR) to CRD at graded pressures (0-60 mmHg) were measured by the number of reflex abdominal contractions. Acute colorectal hypersensitivity was induced by intracolonic infusion of dilute acetic acid. Chronic hypersensitivity was observed in rats following spontaneous resolution of trinitrobenzenesulfonic acid induced colitis. Rats with normosensitive colons served as controls. Tegaserod (0.1-10 mg kg(-1)) caused dose-dependent reduction of the VMR to CRD in control rats and in those with colonic hypersensitivity. 5-HT(4) antagonists reversed the effects of tegaserod in rats with normosensitive colons, and partially inhibited effects in rats with colonic hypersensitivity. Central administration of tegaserod had no inhibitory effect. These results support the assumption that colonic hypersensitivity could be normalized by tegaserod acting, at least in part, through peripheral 5-HT(4) receptors. PMID- 16371087 TI - The CARI guidelines. Nutrition and growth in kidney disease. PMID- 16371108 TI - The CARI guidelines. Cardiovascular risk factors. Lipid-lowering therapy in patients with chronic kidney disease. PMID- 16371109 TI - Melatonin hormone profile in infertile males. AB - Melatonin is a hormone produced by the pineal gland. There is much controversy about its relationship to the male reproductive process. In this study, seminal plasma as well as the serum melatonin levels were studied in different infertile male groups and were correlated with their semen parameters and hormonal levels. One hundred twenty male cases subdivided into six equal groups were consecutively included; fertile normozoospermic men, oligoasthenozoospermia (OA), OA with leucocytospermia, OA with varicocele, non-obstructive azoospermia (NOA) with high serum follicle stimulating hormone (FSH) and NOA with normal FSH. Semen analysis, estimation of melatonin, FSH, testosterone (T) and prolactin (PRL) hormone was carried out. Mean level of serum melatonin was higher than its corresponding seminal concentrations in all investigated groups with a positive correlation between their levels (r = 0.532, p = 0.01). Serum and seminal plasma melatonin levels in all infertile groups were reduced significantly compared with their levels in the fertile group. The lowest concentrations were in OA with leucocytospermia group. Melatonin in both serum and semen demonstrated significant correlation with sperm motility (r = 607, 0.623 respectively, p = 0.01). Serum melatonin correlated positively with serum PRL (r = 0.611, p = 0.01). It may be concluded that melatonin may be involved in the modulation of reproductive neuroendocrine axis in male infertility. Also, low levels of melatonin in semen were observed in infertile groups having reduced sperm motility, leucocytospermia, varicocele and NOA. PMID- 16371110 TI - In utero exposure to persistent organic pollutants in relation to testicular cancer risk. AB - Testicular cancer is the most common tumour type in young men. In Sweden the annual age-adjusted incidence increased significantly by 2.4% during the time period 1984-1993 and during 1994-2003 by 1.4%. Exposure to endocrine disrupting chemicals during the foetal period has been postulated to be a risk factor. In this investigation we studied the concentrations of chlorinated biphenyls (PCBs), p,p'-dichlorodiphenyl-dichloroethylene (pp'-DDE), hexachlorobenzene (HCB), chlordanes and polybrominated diphenylethers (PBDEs) in 58 cases with testicular cancer and 61 age-matched controls. Furthermore, case and control mothers were also asked to participate and 44 case mothers and 45 control mothers agreed. No significant differences were found between cases and controls. Case mothers had in general higher concentrations of these chemicals. For the sum of PCBs an odds ratio (OR) = 3.8, 95% confidence interval (CI) = 1.4-10 was calculated using the median concentration for the controls as cut-off value. For HCB OR = 4.4, CI = 1.7-12 and for PBDE OR = 2.5, 95% CI = 1.02-6.0 were obtained, whereas OR was not significantly increased for pp'-DDE and sum of chlordanes. The cases were born during a period with high concentrations of persistent organic pollutants (POPs) in humans. The decline of the increasing incidence of testicular cancer during recent years may reflect decreasing body burden of certain POPs since the 1980s. PMID- 16371111 TI - Seminal tissue factor revisited. AB - Studies of seminal tissue factor (TF) are few and mostly based on small numbers. Due to the reported lack of factor (F) X in semen, it has been suggested that TF may not have a role in seminal coagulum formation. However, recent identification of a number of haemostatic factors in semen justifies a re-evaluation of its occurrence. Semen specimens were collected from sub-fertile (n = 19), normally fertile (n = 33), semen donors (n = 30) and vasectomized subjects (n = 62), some fractionated into sperm, a prostasome-rich fraction and seminal plasma. Functional and antigenic TF levels were measured and related to conventional fertility parameters. Semen contains high concentration of functional and antigenic TF. Most TF was found in seminal plasma prepared by low-speed centrifugation. When further fractionated by ultracentrifugation much of this may reside in the pellet (prostasomal fraction). It was also detectable on sperm. TF antigen levels were higher in vasectomized subjects than sub-fertile, normally fertile, donor (p = 0.02) and a 'pooled normal semen parameters' (PNSP) stratification (derived from a combination of measurements) (p = 0.06). The sub fertile group showed a wider variation than normal, donor or the PNSP subjects. Seminal TF antigen levels correlated significantly with sperm agglutination (p = 0.03) and abnormal sperm morphology (p = 0.04). Subjects with anti-sperm antibodies also showed high TF antigen levels. In conclusion, semen contains functional and antigenic TF at high concentrations. A full complement of clotting factors probably exists in semen, so some pro-coagulant role for TF should not be excluded. Decreased seminal TF levels appear to be associated with seminal parameters that are known to favour male fertility. PMID- 16371112 TI - Trends in testicular cancer incidence in the Nordic countries, focusing on the recent decrease in Denmark. AB - Testicular cancer is the most frequent malignancy among young men, and there have been steady increases in its incidence in most western countries for many decades. Recently, a decrease was seen in some countries, including Denmark. Here, we report recent trends in testicular cancer incidence in the Nordic countries. We address the hypothesis that the causal factors for testicular cancer in Denmark are related to birth cohort and that non-seminoma and seminoma tumours have a common aetiology. An overall increase in testicular cancer incidence was found in the Nordic countries, corresponding to increases with each consecutive birth cohort in each country. In Denmark, a decline in incidence was observed during the past 5 years, and men born around 1943 and around 1968 showed lower incidences than men born just before or just after these dates. These birth cohort effects were seen both for seminoma and non-seminoma tumours. This descriptive study confirms the hypothesis that birth cohort has a major influence on the incidence pattern of testicular tumours and suggests that seminoma and non seminoma have common aetiological factors. PMID- 16371113 TI - The roe deer as a model for studying seasonal regulation of testis function. AB - In mature male seasonal breeders, the circannual cycles of testicular growth and involution involve significant changes in structure and function of both the tubular and interstitial testicular compartment. Roe deer (Capreolus capreolus) are seasonal breeders with a short defined rutting season from mid-July to mid August and represent a unique non-rodent model for studying testicular functions during the course of a complete reproductive cycle with naturally changing photoperiod. Germ cells and Sertoli cells of the seminiferous tubules and the interstitial Leydig cells all display significant morphological and physiological alterations during the seasonal changes. In contrast to the germ cell population, Sertoli and Leydig cells persist as a numerically constant cell population in the roe deer testis. This report emphasizes the intricate relationship between seasonal cellular differentiation, intratesticular growth factor networks and their impact on the functional dynamics during the seasonal changes in roe deer testis. PMID- 16371114 TI - Influence of obesity on timing of puberty. AB - Timing of normal pubertal maturation has received increased attention over the past several years. Age of menarche showed a dramatic decrease in the first half of the 20th century in Western nations; since 1960, the decrease in age of menarche has slowed and, in some societies, has actually increased. More recently, age of onset of pubertal maturation appears to have decreased, especially in girls. Multiple studies have demonstrated that childhood levels of obesity and ponderosity are associated with earlier menarche in girls. Earlier maturation in boys, however, is associated with lower adiposity and ponderosity. Secular changes in the United States as well as around the world have noted increases in prevalence as well as extent of overweight. Factors contributing to these changes include increases in calories and fast food consumption, decreases in physical activity, and increases in television viewing. There appear to have been selective advantages to lower metabolic rate (the 'thrifty' gene hypothesis), and earlier maturation. However, changes in caloric consumption, caloric density and physical activity, combined with greater lifespan, may allow expression of the adverse consequences of these adaptations, such as the metabolic syndrome or cancers. PMID- 16371116 TI - In-cell Western assay: a new approach to visualize tissue factor in human monocytes. AB - BACKGROUND: Tissue factor (TF) is an integral membrane protein essential for the initiation of the extrinsic pathway of hemostasis. A precise understanding of the TF regulation is still limited and dependent on the availability of methodological tools. Here, we describe a new approach for assessing TF amounts in human mononuclear cells (MNCs) by using the whole blood experimental conditions. AIM: In order to study TF antigen levels in human MNCs, we applied a quantitative immunostaining technique-- in-cell Western (ICW) assay using an Odyssey Infrared Imaging System. METHODS AND RESULTS: The ICW assay of TF in resting or lipopolysaccharide (LPS)-stimulated human MNCs was performed. Several sample preparation conditions were tested, namely the plating of MNCs prior to immunostaining, paraformaldehyde fixation, and an adequate cell number was used in the assay. By the use of recombinant human TF standards, it was possible, for the first time, to measure TF amounts in LPS-stimulated MNCs as 0.09 +/- 0.02 ng and 0.43 +/- 0.15 ng 10(-6) cells of surface and total TF, respectively. The concentrations of TF in resting MNCs, however, were below the detection limit. CONCLUSIONS: A novel TF ICW assay is a reproducible, time- and cost-saving method, which could become useful for studies in the fields of physiology and pathophysiology of human hemostasis. PMID- 16371117 TI - Genetic influence on thrombotic risk markers in the elderly--a Danish twin study. AB - OBJECTIVE: Several hemostatic variables are identified as cardiovascular risk markers. In young and middle-aged individuals, plasma concentrations of these variables are partly determined by genetic factors. The genetic contribution to cardiovascular disease (CVD) decreases with increasing age, and it is therefore important to determine the heritability of hemostasis also in the elderly. METHODS: The heritability of plasma levels of factor VII, fibrinogen, tissue factor, tissue factor pathway inhibitor, von Willebrand factor, thrombin activatable fibrinolysis inhibitor (TAFI), and D-dimer was determined in 130 monozygotic and 155 dizygotic same-sex twin pairs, aged 73-94 years, who participated in the Longitudinal Study of Aging of Danish Twins. Furthermore, we determined the influence of promoter polymorphisms in corresponding genes on the plasma level variation. RESULTS: Genetic factors accounted for 33% (D-dimer) to 71% (TAFI) of the variation in plasma levels. Polymorphisms were associated with concentrations of FVII and TAFI in sib-pair based analyses, but in linkage analyses the polymorphisms did not explain a significant part of the genetic variation for any of the variables. CONCLUSIONS: Concentrations of hemostatic variables have a substantial genetic variation in the elderly, but in this study the promoter polymorphisms only explained a minimal part of this variation. PMID- 16371118 TI - Soluble tissue factor is a candidate marker for progression of microvascular disease in patients with Type 2 diabetes. AB - OBJECTIVE: To determine the relationship between abnormalities in blood coagulation and prevalent or incident cardiovascular complications in Type 2 diabetes. DESIGN AND METHODS: Prospective cohort study of 128 patients with Type 2 diabetes in whom blood samples were collected at baseline and after 1 year of follow-up. All cardiovascular complications at baseline and follow-up were recorded. Forty-three healthy, age-matched subjects served as a control group. RESULTS: Logistic analysis revealed an independent relationship between soluble tissue factor (TF) and microvascular disease [per pg mL(-1) TF: Exp(B) = 1.008; CI(95%)1.002-1.014], or neurogenic disease [Exp(B) = 1.006; CI(95%)1.001-1.011]. The highest levels of soluble TF were observed in patients with microvascular and neurogenic disease (P < 0.001). Patients with Type 2 diabetes having a soluble TF concentration >300 pg mL(-1) are at a 15-fold higher risk for the presence of microvascular disease and at a 10-fold higher risk for the presence of neurogenic disease compared with the patients with concentrations below 100 pg mL(-1). Soluble TF was correlated with tissue type plasminogen activator, von Willebrand factor antigen, systolic blood pressure and age. Levels of F1' + 2, D-dimer, FVIII activity, t-PA and vWFag were not different among patients with micro-, macro- or neurogenic complications compared with patients without those complications. Forty-eight new micro-, macro- and/or neurogenic complications were diagnosed after 1 year follow-up. With the exception of higher F1 + 2 levels after 1 year all other markers remained unchanged. A trend toward higher soluble TF levels was observed in patients with new microvascular events (P = 0.056). CONCLUSIONS: Soluble TF is associated with existing microvascular and neurogenic complications in patients with Type 2 diabetes and is a candidate marker for progression of microvascular disease. PMID- 16371120 TI - Did Jesus Christ die of pulmonary embolism? PMID- 16371119 TI - High post-treatment platelet reactivity identified low-responders to dual antiplatelet therapy at increased risk of recurrent cardiovascular events after stenting for acute coronary syndrome. AB - BACKGROUND AND OBJECTIVES: Low response to antiplatelet therapy may be a risk factor for the development of ischemic complications in patients with non-ST segment elevation acute coronary syndrome (NSTE ACS) undergoing coronary stenting. METHODS: We prospectively studied the platelet response to both clopidogrel and aspirin in 106 NSTE ACS consecutive patients undergoing percutaneous coronary intervention (PCI) with stenting. A single post-treatment blood sample was obtained just before PCI and analyzed by platelet aggregometry using both ADP and arachidonic acid (AA) as agonists to explore the responses to clopidogrel and aspirin, respectively. Patients were divided into quartiles according to the ADP or AA induced maximal intensity of platelet aggregation. Patients of the highest quartile (quartile 4) were defined as the 'low responders'. RESULTS: Twelve recurrent cardiovascular (CV) events occurred during the 1-month follow-up. Clinical outcome was significantly associated with platelet response to clopidogrel [Quartile 4 vs. 1, 2, 3: OR (95% CI) 22.4 (4.6 109)]. Low platelet response to aspirin was significantly correlated with clopidogrel low response (P = 0.003) but contributed less to CV events [OR (95%CI): 5.76 (1.54-35.61)]. CONCLUSIONS: A post-treatment ADP-induced platelet aggregation performed just before PCI identifies low responders to dual antiplatelet therapy with an increased risk of recurrent CV events. PMID- 16371122 TI - A concept analysis: the grieving process for nurses. AB - The concept of the grieving process has been explored extensively in families losing a loved one or in a patient grieving over a terminal diagnosis. The patients and families live through this experience one time. What about the nurse who lives it several times a week by caring for these patients and families? How does a nurse grieve? Little publication and research have been done surrounding the grieving process for nurses. This is a concept analysis that clarifies the grieving process for nurses. Clarifying this process will enable further development of nursing research and education, ultimately benefiting nursing practice and retention. PMID- 16371123 TI - To be or not to be: terminal illness in film and in life. PMID- 16371124 TI - The value of mentorship within nursing organizations. AB - PROBLEM: Hospitals are facing nurse retention challenges in the new millennium. Nurses are abandoning the bedside because of job dissatisfaction. Mentorship programs should be developed to enhance nursing satisfaction and thus improve retention and ultimately patient outcomes. FINDINGS: Mentorship training in hospitals and the support of formal mentorship programs for new nurses are ways in which hospitals can meet this challenge. CONCLUSIONS: This paper offers an overview of recent research on the value mentoring has for participants and healthcare organizations. The components of successful mentorship programs are discussed, and guidelines for implementation are recommended. PMID- 16371125 TI - What is in a name: the case of nursing. AB - The purpose of this paper is to create a debate on whether the name "nursing" has any influence on how the nursing profession is perceived today by other healthcare professionals and the general public the nursing profession serves. A quantum leap is being suggested by the authors, as only a paradigm shift could change the world's mindset on nursing and its recognition as a profession. A change in name is what the authors see as a way of changing the unflattering perception of nurses and enhancing its status to the level of respectability of other healthcare professions. PMID- 16371126 TI - Practice week at a glance: nursing 40 years ago and today. PMID- 16371128 TI - Visualization of the interaction between archaeal DNA polymerase and uracil containing DNA by atomic force microscopy. AB - Deamination of cytosine to uracil is a hydrolytic reaction that is greatly accelerated at high temperatures. The resulting uracil pairs with adenine during DNA replication, thereby inducing G:C to A:T transitions in the progeny. Interestingly, B-family DNA polymerases from hyperthermophilic Archaea recognize the presence of uracil in DNA and stall DNA synthesis. To better understand the recognition mechanism, the binding modes of DNA polymerase B1 of Sulfolobus solfataricus (Pol B1) to uracil-containing DNA were examined by gel mobility shift assays and atomic force microscopy. Although PolB1 per se specifically binds to uracil-containing single-stranded DNA, the binding efficiency was substantially enhanced by the initiation of DNA synthesis. Analysis by the atomic force microscopy showed a number of double-stranded DNA (dsDNA) in the products of DNA synthesis. The generation of ds DNA was significantly inhibited, however, by the presence of template uracil, and intermediates where monomeric forms of Pol B1 appeared to bind to uracil-containing DNA were observed. These results suggest that Pol B1 more efficiently recognizes uracil in DNA during DNA synthesis rather than during random diffusion in solution, and that single molecules of Pol B1 bind to template uracil and stall DNA synthesis. PMID- 16371129 TI - Distinct modes of DNA damage response in S. pombe G0 and vegetative cells. AB - Upon nitrogen-starvation, mostly G2 vegetative (VE) fission yeast cells promote two rounds of division and enter the G0 state with 1C DNA via an uncommitted G1. Whilst G0 cells are permanently arrested, they keep viability through recycling the intracellular nitrogen. We here show that, whilst the DNA damages are efficiently repaired in G0 cells, neither Chk1 activation nor Cdc2 implication for Crb2 (53BP1 like) do not occur. ATR-like Rad3 and non-hyperphosphorylated Crb2 participate the repair processes in G0 cells that are more sensitive to UV and gamma-ray than in VE cells. The sensitivity like in VE cells is restored after replication in the nitrogen-replenished medium, suggesting that the damage hyper-sensitive nature of G0 cells is due to the error-prone repair for single DNA duplex chromosome. The double-strand break (DSB) repair in G0 cells required Pku80, one of non-homologous end joining (NHEJ) proteins. S. pombe G0 cells upon DNA damages thus respond distinctively from VE cells in regard with regulation of checkpoint proteins and the mode of repair that is dependent upon the use of NHEJ. PMID- 16371130 TI - Loss of RanGEF/Pim1 activity abolishes the orchestration of Ran-mediated mitotic cellular events in S. pombe. AB - RCC1, a conserved chromosomal protein with a seven-bladed propeller is a GDP/GTP nucleotide exchange factor for RanGTPase that mediates various cellular events. We isolated 16 temperature-sensitive (ts) mutants of S. pombeRCC1-homolog, pim1+, by error-prone PCR. Five pim1(ts) mutants had a single mutation. The obtained pim1(ts) mutations and previously reported mutations were localized on similar sites in seven RCC1 repeats. Those mutations resulted in a reduced binding of Pim1 with Spi1. All pim1(ts) mutants showed a defect in nucleocytoplasmic protein transports, whereas the majority of them showed a normal mRNA export. In all pim1(ts) examined, chromosomal DNA replication was completed. However, mitotic spindle formation was abrogated, the septum was formed being uncoupled with nuclear division and abnormally widened, thus resulting in chromosomal DNA mis segregation and the accumulation of enucleated cells. As a result, a defect of RanGEF/Pim1 abolished the orchestration of sequential mitotic events, spindle formation, septation and cytokinesis that are essential to produce two identical daughter cells. PMID- 16371131 TI - Direct binding of TReP-132 with TdT results in reduction of TdT activity. AB - N regions at the junction of V, D and J DNA segments are synthesized with large protein complexes including terminal deoxynucleotidyltransferase (TdT) during V(D)J recombination in B- or T-cells. TdT directly binds to TdIF1, TdIF2, PCNA and the Ku70/86 heterodimer. Using a yeast two-hybrid system, we isolated a cDNA clone encoding the gene for TReP-132, which is involved in P450scc gene expression in steroid-hormone-producing cells or lymphoid cells. Interaction between TReP-132 and TdIF1 was confirmed by pull-down assay and immunoprecipitation assay using specific antibodies against TReP-132 both in vitro and in vivo. TdT also directly bound to TReP-132 through its confined N terminal region. Furthermore, the co-expression of TdIF1 and TReP-132 or TdT and TReP-132 in COS7 cells showed that these proteins are co-localized within the nucleus. TReP-132 reduces TdT activity to 2.5% of its maximum value in the in vitro assay system using double-stranded DNA with a 3' protrusion as a primer. These findings suggest that TdT synthesizes N region under a negative control of TReP-132 during V(D)J recombination. PMID- 16371132 TI - Yeast unfolded protein response pathway regulates expression of genes for anti oxidative stress and for cell surface proteins. AB - The unfolded protein response (UPR) is a cellular protective event against endoplasmic reticulum (ER) stress. In the yeast UPR signaling pathway, the ER located transmembrane protein Ire1 promotes splicing of the HAC1 premRNA (HAC1(u)) to produce the translatable transcription factor mRNA (HAC1i). We generated a HAC1i gene-bearing strain, in which the UPR pathway was constitutively activated, and compared its gene expression profile with that of a Deltaire1 HAC1u strain using cDNA microarray technology. Comparison of the gene expression profile was also performed between non-stressed wild-type cells and those exposed to ER stress. Genes for which the expression level was significantly changed in both of these experiments were categorized as targets of the Ire1-HAC1 signaling pathway. This analysis revealed that in addition to the previously known UPR targets, some anti-oxidative stress genes were up-regulated by the Ire1-HAC1 pathway, possibly in order to reduce reactive oxygen species produced during the cellular response to ER stress. Moreover, we categorized 15 genes as those down-regulated by the UPR, most of which seem to encode cell surface or extracellular proteins. This UPR-mediated gene repression may alleviate the load of client proteins targeted to the ER. PMID- 16371133 TI - Developmental expression patterns of testicular olfactory receptor genes during mouse spermatogenesis. AB - A subset of olfactory receptors (ORs) is expressed in mammalian male germ cells. Recent studies on human and mouse sperm have suggested that calcium signaling via a testicular OR regulates sperm flagellar motility. However, it remains to be determined at what stages testicular ORs are expressed during spermatogenesis and whether each germ cell expresses one or multiple ORs. Here we examined the developmental expression profiles of several mouse testicular OR genes using an in situ hybridization technique at the cellular level. We found that OR transcripts in the spermatogenic cells are expressed in three developmental stages: late pachyten spermatocytes, early round spermatids, or late round spermatids. The OR mRNAs were condensed in a single dot-like structure within the nuclei of a subpopulation of spermatogenic cells. Double-fluorescent in situ hybridization revealed that some cells contained two dot-like signals derived from transcripts of two different ORs, suggesting that single spermatogenic cells could express more than one OR. One cell-multiple OR gene expression combined with variability in expression appears to result in heterogeneity in the repertoire of ORs expressed by individual spermatogenic cells. Although the functional consequence of heterogeneous OR expression awaits development of a methodology for characterizing OR proteins, our observations give insights into OR gene expression as well as OR function(s) in spermatogenic cells. PMID- 16371134 TI - Reconstitution of gamma-secretase by truncated presenilin (PS) fragments revealed that PS C-terminal transmembrane domain is critical for formation of gamma secretase complex. AB - The presenilin (PS) complex, including PS, nicastrin (NCT), APH-1 and PEN-2, is essential for gamma-secretase activity. Previously, the PS C-terminal tail was shown to be essential for gamma-secretase activity. Here, to further understand the precise mechanism underlying the activation of gamma-secretase regulated by PS cofactors, we focused on the role of the PS1 C-terminal region including transmembrane domain (TM) 8 in gamma-secretase activity. For this purpose, we co expressed C-terminally truncated PS1 (PS1DeltaC) completely lacking gamma secretase activity and the PS1 C-terminal short fragment in PS-null cells, because the successful reconstitution of gamma-secretase activity in PS-null cells by the co-expression of PS1DeltaC and the PS1 C-terminal short fragment would allow us to investigate the role of the PS1 C-terminal region in gamma secretase activity. We found that the exogenous expression of the PS1 C-terminal short fragment with NCT and APH-1 completely rescued a defect of the gamma secretase activity of PS1DeltaC in PS-null cells. With this reconstitution system, we demonstrate that both TM8 and the PS1 C-terminal seven-amino-acid residue tail are involved in the formation of the active gamma-secretase complex via the assembly of PS1 with NCT and APH-1. PMID- 16371135 TI - Evaluation of somatic alterations of Pcdh-alpha transcripts in the brain by cDNA analysis without PCR. PMID- 16371136 TI - How color enhances visual memory for natural scenes. AB - We offer a framework for understanding how color operates to improve visual memory for images of the natural environment, and we present an extensive data set that quantifies the contribution of color in the encoding and recognition phases. Using a continuous recognition task with colored and monochrome gray scale images of natural scenes at short exposure durations, we found that color enhances recognition memory by conferring an advantage during encoding and by strengthening the encoding-specificity effect. Furthermore, because the pattern of performance was similar at all exposure durations, and because form and color are processed in different areas of cortex, the results imply that color must be bound as an integral part of the representation at the earliest stages of processing. PMID- 16371137 TI - An asymmetry between memory encoding and retrieval. Revelation, generation, and transfer-appropriate processing. AB - Transfer-appropriate-processing accounts of memory emphasize the similarity of encoding and retrieval processes, and imply that experimental manipulations should have similar effects on encoding and retrieval. Exceptions to this expectation are thus of great interest, but extant exceptions (produced by studies using divided attention, alcohol, and benzodiazepines) are debatable, single dissociations between encoding and retrieval. The present experiments demonstrate a reversed dissociation, in which the same variable produced opposite effects when implemented at encoding and retrieval. At encoding, participants either solved anagrams of study words or read intact study words. At retrieval, participants likewise solved anagrams or read intact words prior to making recognition memory judgments. Compared with reading intact words, solving anagrams at encoding enhanced later recognition accuracy, whereas solving anagrams at test impaired accuracy. These results were obtained with old/new decisions (Experiment 1) and with confidence ratings (Experiment 2). PMID- 16371138 TI - Activation of face expertise and the inversion effect. AB - We used a contextual priming paradigm to examine top-down influences on the face inversion effect. Adult participants were primed with either faces or Chinese characters and then tested on ambiguous figures that could be perceived as either faces or Chinese characters, dependent on the priming condition. The ambiguous figures differed from one another in their configural information, which is crucial for processing faces but not Chinese characters. The inversion effect was observed in the face-priming condition, but not in the character-priming condition. The present results provide the first direct evidence that top-down activation of the face-processing expertise system plays a crucial role in the face-inversion effect. PMID- 16371139 TI - Doing is for thinking! Stereotype activation by stereotypic movements. AB - Three studies demonstrate that stereotypic movements activate the corresponding stereotype. In Study 1, participants who were unobtrusively induced to move in the portly manner that is stereotypic of overweight people subsequently ascribed more overweight-stereotypic characteristics to an ambiguous target person than did control participants. In Study 2, participants who were unobtrusively induced to move in the slow manner that is stereotypic of elderly people subsequently ascribed more elderly-stereotypic characteristics to a target than did control participants. In Study 3, participants who were induced to move slowly were faster than control participants to respond to elderly-stereotypic words in a lexical decision task. Using three different movement inductions, two different stereotypes, and two classic measures of stereotype activation, these studies converge in demonstrating that stereotypes may be activated by stereotypic movements. PMID- 16371140 TI - Do approach-avoidance actions create attitudes? AB - Do approach-avoidance actions create attitudes? Prior influential studies suggested that rudimentary attitudes could be established by simply pairing novel stimuli (Chinese ideographs) with arm flexion (approach) or arm extension (avoidance). In three experiments, we found that approach-avoidance actions alone were insufficient to account for such effects. Instead, we found that these affective influences resulted from the interaction of these actions with a priori differences in stimulus valence. Thus, with negative stimuli, the effect of extension on attitude was more positive than the effect of flexion. Experiment 2 demonstrated that the affect from motivationally compatible or incompatible action can also influence task evaluations. A final experiment, using Chinese ideographs from the original studies, confirmed these findings. Both approach and avoidance actions led to more positive evaluations of the ideographs when the actions were motivationally compatible with the prior valence of the ideographs. The attitudinal impact of approach-avoidance action thus reflects its situated meaning, which depends on the valence of stimuli being approached or avoided. PMID- 16371141 TI - Alcohol and aggression without consumption. Alcohol cues, aggressive thoughts, and hostile perception bias. AB - Researchers and the lay public have long known of a link between alcohol and aggression. However, whether this link results from alcohol's pharmacological effects or is merely an artifact of the belief that alcohol has been consumed (i.e., placebo effect) has been debated. The current experiments examined the propensity for alcohol-related cues to elicit aggressive thoughts and hostile perceptions in the absence of alcohol or placebo consumption. In Experiment 1, participants made faster lexical decisions concerning aggression-related words following alcohol-related primes compared with neutral primes. In Experiment 2, participants who first were exposed to alcohol advertisements subsequently rated the behavior of a target person as more hostile than participants who initially viewed control advertisements. Furthermore, this effect was largest among participants who most strongly associated alcohol and aggression. Findings are discussed in terms of semantic network theory and links in memory between alcohol and its anticipated effects. PMID- 16371142 TI - Misperceptions in intergroup conflict. Disagreeing about what we disagree about. AB - Two studies examined misperceptions of disagreement in partisan social conflicts, namely, in the debates over abortion (Study 1) and politics (Study 2). We observed that partisans tend to exaggerate differences of opinion with their adversaries. Further, we found that perceptions of disagreement were more pronounced for values that were central to the perceiver's own ideology than for values that were central to the ideology of the perceiver's adversaries. To the extent that partisans assumed disagreement concerning personally important values, they were also inaccurate in perceiving their adversaries' actual opinions. Discussion focuses on the cognitive mechanisms underlying misperceptions of disagreement and strategies for reducing intergroup conflict. PMID- 16371143 TI - Is high sex drive associated with increased sexual attraction to both sexes? It depends on whether you are male or female. AB - If sex drive is a generalized energizer of sexual behaviors, then high sex drive should increase an individual's sexual attraction to both men and women. If sex drive energizes only dominant sexual responses, however, then high sex drive should selectively increase attraction to men or to women, but not to both, depending on the individual's sexual orientation. Data from three studies assessing a total of 3,645 participants show that for most women, high sex drive is associated with increased sexual attraction to both men and women. For men, however, high sex drive is associated with increased sexual attraction to only one sex or the other, depending on the individual's sexual orientation. These results suggest that the correlates of sex drive and the organization of sexual orientation are different for women and men. PMID- 16371144 TI - The development of implicit attitudes. Evidence of race evaluations from ages 6 and 10 and adulthood. AB - To understand the origin and development of implicit attitudes, we measured race attitudes in White American 6-year-olds, 10-year-olds, and adults by first developing a child-oriented version of the Implicit Association Test (Child IAT). Remarkably, implicit pro-White/anti-Black bias was evident even in the youngest group, with self-reported attitudes revealing bias in the same direction. In 10 year-olds and adults, the same magnitude of implicit race bias was observed, although self-reported race attitudes became substantially less biased in older children and vanished entirely in adults, who self-reported equally favorable attitudes toward Whites and Blacks. These data are the first to show an asymmetry in the development of implicit and explicit race attitudes, with explicit attitudes becoming more egalitarian and implicit attitudes remaining stable and favoring the in-group across development. We offer a tentative suggestion that mean levels of implicit and explicit attitudes diverge around age 10. PMID- 16371145 TI - Neural markers of categorization in 6-month-old infants. AB - Little is known of the neural processes that underlie concept-formation abilities in human infants. We investigated category-learning processes in infants both by using a common behavioral measure and by recording the brain's electrical activity (event-related potentials, or ERPs). ERPs were recorded while 6-month olds viewed cat images during training, followed by novel cat images interspersed with novel dog images during test. The data indicate that distinct neural signals correspond with learning of a category presented during familiarization, preferential responding to a novel category, and representation of category exemplars at multiple levels of inclusiveness. The results suggest that fundamental components of the neural architecture supporting object categorization are functional within the first half-year of postnatal life, before infants acquire language and young children engage in formal learning of semantic categories. The findings are discussed in terms of their implications for models of category learning and development. PMID- 16371146 TI - Rethinking speed theories of cognitive development. Increasing the rate of recall without affecting accuracy. AB - Researchers have suggested that developmental improvements in immediate recall stem from increases in the speed of mental processes. However, that inference has depended on evidence from correlation, regression, and structural equation modeling. We provide counter-examples in two experiments in which the speed of spoken recall was manipulated. In one experiment, second-grade children and adults recalled lists of digits more quickly than usual when the lists were presented at a rapid rate of two items per second. In a second experiment, children received lists at a rate of one item per second; half the children were trained (successfully) to speak their responses more quickly than usual, at a rate similar to adults' usual rate. Recall accuracy was completely unaffected by either of these response-speed manipulations. Thus, although response rate is a strong marker of an individual's maturational level, it does not appear to determine the accuracy of immediate recall. These results have important methodological and theoretical implications for human development. PMID- 16371147 TI - The development of executive functioning and theory of mind. A comparison of Chinese and U.S. preschoolers. AB - Preschoolers' theory-of-mind development follows a similar age trajectory across many cultures. To determine whether these similarities are related to similar underlying ontogenetic processes, we examined whether the relation between theory of mind and executive function commonly found among U.S. preschoolers is also present among Chinese preschoolers. Preschoolers from Beijing, China (N= 109), were administered theory-of-mind and executive-functioning tasks, and their performance was compared with that of a previously studied sample of U.S. preschoolers (N= 107). The Chinese preschoolers outperformed their U.S. counterparts on all measures of executive functioning, but were not similarly advanced in theory-of-mind reasoning. Nonetheless, individual differences in executive functioning predicted theory of mind for children in both cultures. Thus, the relation between executive functioning and theory of mind is robust across two disparate cultures. These findings shed light on why executive functioning is important for theory-of-mind development. PMID- 16371148 TI - Purchases of food in youth. Influence of price and income. AB - One way to increase choice of healthy over unhealthy behaviors is to increase the cost of less healthy alternatives or reduce the cost of healthier alternatives. The influence of price on purchases of healthy and unhealthy foods was evaluated in two laboratory experiments. In Experiment 1, thirty-two 10- to 12-year-old youth were given $5.00 and allowed to purchase multiple portions of a healthy food (fruit or vegetable) and a less healthy food (higher-fat snack). The price of one type of food varied from $0.50 to $2.50, while the price of the other type was held at $1.00. Increasing the price of a type of food reduced purchases of that type of food, but did not lead to substitution with the alternative type of food. In Experiment 2, twenty 10- to 14-year-old youth were given $1.00, $3.00, and $5.00 to purchase healthy and unhealthy foods. The price of each food was raised and lowered by 25% and 50%. Raising the price of healthy or unhealthy foods resulted in decreased purchases of those foods, and income available interacted with price to predict the pattern of substitution of alternative foods. These results show the potential for controlled laboratory studies of price and food purchases, and show that the substitution of healthier for unhealthy food is related to available money. PMID- 16371149 TI - A Global Workspace perspective on mental disorders. AB - BACKGROUND: Recent developments in Global Workspace theory suggest that human consciousness can suffer interpenetrating dysfunctions of mutual and reciprocal interaction with embedding environments which will have early onset and often insidious staged developmental progression, possibly according to a cancer model, in which a set of long-evolved control strategies progressively fails. METHODS AND RESULTS: A rate distortion argument implies that, if an external information source carries a damaging 'message', then sufficient exposure to it, particularly during critical developmental periods, is sure to write a sufficiently accurate image of it on mind and body in a punctuated manner so as to initiate or promote similarly progressively punctuated developmental disorder, in essence either a staged failure affecting large-scale brain connectivity, which is the sine qua non of human consciousness, or else damaging the ability of embedding goal contexts to contain conscious dynamics. CONCLUSION: The key intervention, at the population level, is clearly to limit exposure to factors triggering developmental disorders, a question of proper environmental sanitation, in a large sense, primarily a matter of social justice which has long been known to be determined almost entirely by the interactions of cultural trajectory, group power relations, and economic structure, with public policy. Intervention at the individual level appears limited to triggering or extending periods of remission, representing reestablishment of an extensive, but largely unexplored, spectrum of evolved control strategies, in contrast with the far better-understood case of cancer. PMID- 16371150 TI - Intestinal invasion and disseminated disease associated with Penicillium chrysogenum. AB - BACKGROUND: Penicillium sp., other than P. marneffei, is an unusual cause of invasive disease. These organisms are often identified in immunosuppressed patients, either due to human immunodeficiency virus or from immunosuppressant medications post-transplantation. They are a rarely identified cause of infection in immunocompetent hosts. CASE PRESENTATION: A 51 year old African-American female presented with an acute abdomen and underwent an exploratory laparotomy which revealed an incarcerated peristomal hernia. Her postoperative course was complicated by severe sepsis syndrome with respiratory failure, hypotension, leukocytosis, and DIC. On postoperative day 9 she was found to have an anastamotic breakdown. Pathology from the second surgery showed transmural ischemic necrosis with angioinvasion of a fungal organism. Fungal blood cultures were positive for Penicillium chrysogenum and the patient completed a 6 week course of amphotericin B lipid complex, followed by an extended course oral intraconazole. She was discharged to a nursing home without evidence of recurrent infection. DISCUSSION: Penicillium chrysogenum is a rare cause of infection in immunocompetent patients. Diagnosis can be difficult, but Penicillium sp. grows rapidly on routine fungal cultures. Prognosis remains very poor, but aggressive treatment is essential, including surgical debridement and the removal of foci of infection along with the use of amphotericin B. The clinical utility of newer antifungal agents remains to be determined. PMID- 16371151 TI - Kinetics of hepatitis C virus RNA load during pegylated interferon alpha-2a and ribavirin treatment in naive genotype 1 patients. AB - BACKGROUND: Pegylated interferon given for 24 or 48 weeks constitutes the most effective initial therapy for the treatment of chronic hepatitis C. It has been shown that viral load at week 2 appears the best time for predicting response to treatment. The objectives of this study were to assess whether the hepatitis C virus (HCV) RNA viral decline is predictive of sustained virological response (SVR) and to determine the best time for predicting complete response in our cohort of naive patients treated with pegylated interferon alpha-2a (Peg-IFN alpha-2a) and ribavirin. RESULTS: Twenty patients treated with Peg-IFN alpha-2a and ribavirin for 48 weeks were studied. Six months after the end of treatment, a SVR (negative HCV RNA measured by PCR six months after the end of therapy) was obtained in 9 patients. Samples were obtained before and at week 2, 4, 8, and 12. At the end of week 2, viral load decreased more than 1.39 log in 8 out of the 9 patients with SVR and in 1 out of the 11 other patients. When we considered the viral load reduction from baseline to each week of treatment, week 2 appeared to be the best point time for predicting SVR, with a sensitivity of 91% (95%CI: 59;99), a specificity of 89% (52;98), a positive predictive value of 91% (59;99) and a negative predictive value of 89% (57;98). CONCLUSION: During treatment with Peg-IFN alpha-2a plus ribavirin in genotype 1 patients, when the main objective of the treatment is viral eradication, viral kinetics showed that week 2 appeared to be the best time point for predicting SVR. Our results must be further confirmed on a larger cohort. PMID- 16371153 TI - Chaperone use during intimate examinations in primary care: postal survey of family physicians. AB - BACKGROUND: Physicians have long been advised to have a third party present during certain parts of a physical examination; however, little is known about the frequency of chaperone use for those specific intimate examinations regularly performed in primary care. We aimed to determine the frequency of chaperone use among family physicians across a variety of intimate physical examinations for both male and female patients, and also to identify the factors associated with chaperone use. METHODS: Questionnaires were mailed to a randomly selected sample of 500 Ontario members of the College of Family Physicians of Canada. Participants were asked about their use of chaperones when performing a variety of intimate examinations, namely female pelvic, breast, and rectal exams and male genital and rectal exams. RESULTS: 276 of 500 were returned (56%), of which 257 were useable. Chaperones were more commonly used with female patients than with males (t = 9.09 [df = 249], p < 0.001), with the female pelvic exam being the most likely of the five exams to be attended by a chaperone (53%). As well, male physicians were more likely to use chaperones for examination of female patients than were female physicians for the examination of male patients. Logistic regression analyses identified two independent factors--sex of physician and availability of a nurse--that were significantly associated with chaperone use. For female pelvic exam, male physicians were significantly more likely to report using a chaperone (adjusted Odds Ratio [OR] 40.62, 95% confidence interval [CI] 16.91-97.52). Likewise, having a nurse available also significantly increased the likelihood of a chaperone being used (adjusted OR 6.92, 95% CI 2.74-17.46). This pattern of results was consistent across the other four exams. Approximately two thirds of respondents reported using nurses as chaperones, 15% cited the use of other office staff, and 10% relied on the presence of a family member. CONCLUSION: Clinical practice concerning the use of chaperones during intimate exams continues to be discordant with the recommendations of medical associations and medico-legal societies. Chaperones are used by only a minority of Ontario family physicians. Chaperone use is higher for examinations of female patients than of male patients and is highest for female pelvic exams. The availability of a nurse in the clinic to act as a chaperone is associated with more frequent use of chaperones. PMID- 16371154 TI - A randomised controlled trial of the effects of albendazole in pregnancy on maternal responses to mycobacterial antigens and infant responses to Bacille Calmette-Guerin (BCG) immunisation [ISRCTN32849447]. AB - BACKGROUND: Maternal schistosomiasis and filariasis have been shown to influence infant responses to neonatal bacille Calmette-Guerin (BCG) immunisation but the effects of maternal hookworm, and of de-worming in pregnancy, are unknown. METHODS: In Entebbe, Uganda, we conducted a randomised, double-blind, placebo controlled trial of a single dose of 400 mg of albendazole in the second trimester of pregnancy. Neonates received BCG. Interferon-gamma (IFN-gamma) and interleukin (IL)-5 responses to a mycobacterial antigen (crude culture filtrate proteins (CFP) of Mycobacterium tuberculosis) were measured in a whole blood assay. We analysed results for binary variables using chi2 tests and logistic regression. We analysed continuous variables using Wilcoxon's tests. RESULTS: Maternal hookworm was associated with reduced maternal IFN-gamma responses to CFP (adjusted odds ratio for IFN-gamma > median response: 0.14 (95% confidence interval 0.02-0.83, p = 0.021). Conversely, maternal hookworm was associated with subsequent increased IFN-gamma responses in their one-year-old infants (adjusted OR 17.65 (1.20-258.66; p = 0.013)). Maternal albendazole tended to reduce these effects. CONCLUSION: Untreated hookworm infection in pregnancy was associated with reduced maternal IFN-gamma responses to mycobacterial antigens, but increased responses in their infants one year after BCG immunisation. The mechanisms of these effects, and their implications for protective immunity remain, to be determined. PMID- 16371155 TI - Prevalence of overweight in children and adolescents with attention deficit hyperactivity disorder and autism spectrum disorders: a chart review. AB - BACKGROUND: The condition of obesity has become a significant public health problem in the United States. In children and adolescents, the prevalence of overweight has tripled in the last 20 years, with approximately 16.0% of children ages 6-19, and 10.3% of 2-5 year olds being considered overweight. Considerable research is underway to understand obesity in the general pediatric population, however little research is available on the prevalence of obesity in children with developmental disorders. The purpose of our study was to determine the prevalence of overweight among a clinical population of children diagnosed with attention deficit hyperactivity disorder (ADHD) and autism spectrum disorders (ASD). METHODS: Retrospective chart review of 140 charts of children ages 3-18 years seen between 1992 and 2003 at a tertiary care clinic that specializes in the evaluation and treatment of children with developmental, behavioral, and cognitive disorders. Diagnostic, medical, and demographic information was extracted from the charts. Primary diagnoses of either ADHD or ASD were recorded, as was information on race/ethnicity, age, gender, height, and weight. Information was also collected on medications that the child was taking. Body mass index (BMI) was calculated from measures of height and weight recorded in the child's chart. The Center for Disease Control's BMI growth reference was used to determine an age- and gender-specific BMI z-score for the children. RESULTS: The prevalence of at-risk-for-overweight (BMI > 85th%ile) and overweight (BMI > 95th%ile) was 29% and 17.3% respectively in children with ADHD. Although the prevalence appeared highest in the 2-5 year old group (42.9%ile), differences among age groups were not statistically significant. Prevalence did not differ between boys and girls or across age groups (all p > 0.05). For children with ASD, the overall prevalence of at-risk-for-overweight was 35.7% and prevalence of overweight was 19%. CONCLUSION: When compared to an age-matched reference population (NHANES 1999-2002), our estimates indicate that children with ADHD and with ASD have a prevalence of overweight that is similar to children in the general population. PMID- 16371156 TI - Relevance of human metapneumovirus in exacerbations of COPD. AB - BACKGROUND AND METHODS: Human metapneumovirus (hMPV) is a recently discovered respiratory virus associated with bronchiolitis, pneumonia, croup and exacerbations of asthma. Since respiratory viruses are frequently detected in patients with acute exacerbations of COPD (AE-COPD) it was our aim to investigate the frequency of hMPV detection in a prospective cohort of hospitalized patients with AE-COPD compared to patients with stable COPD and to smokers without by means of quantitative real-time RT-PCR. RESULTS: We analysed nasal lavage and induced sputum of 130 patients with AE-COPD, 65 patients with stable COPD and 34 smokers without COPD. HMPV was detected in 3/130 (2.3%) AE-COPD patients with a mean of 6.5 x 10(5) viral copies/ml in nasal lavage and 1.88 x 10(5) viral copies/ml in induced sputum. It was not found in patients with stable COPD or smokers without COPD. CONCLUSION: HMPV is only found in a very small number of patients with AE-COPD. However it should be considered as a further possible viral trigger of AE-COPD because asymptomatic carriage is unlikely. PMID- 16371157 TI - Antimicrobial activities of Eugenol and Cinnamaldehyde against the human gastric pathogen Helicobacter pylori. AB - BACKGROUND: Eradication of Helicobacter pylori is an important objective in overcoming gastric diseases. Many regimens are currently available but none of them could achieve 100% success in eradication. Eugenol and cinnamaldehyde that are commonly used in various food preparations are known to possess antimicrobial activity against a wide spectrum of bacteria. AIM: The present study was performed to assess the in vitro effects of eugenol and cinnamaldehyde against indigenous and standard H. pylori strains, their minimum inhibitory concentrations (MICs) and time course lethal effects at various pH. METHODS: A total of 31 strains (29 indigenous and one standard strain of H. pylori ATCC 26695, one strain of E. coli NCIM 2089) were screened. Agar dilution method was used for the determination of drug sensitivity patterns of isolates to the commonly used antibiotics and broth dilution method for the test compounds. RESULTS: Eugenol and cinnamaldehyde inhibited the growth of all the 30 H. pylori strains tested, at a concentration of 2 mug/ml, in the 9th and 12th hours of incubation respectively. At acidic pH, increased activity was observed for both the compounds. Furthermore, the organism did not develop any resistance towards these compounds even after 10 passages grown at sub-inhibitory concentrations. CONCLUSION: These results indicate that the two bioactive compounds we tested may prevent H. pylori growth in vitro, without acquiring any resistance. PMID- 16371158 TI - Alterations of renal phenotype and gene expression profiles due to protein overload in NOD-related mouse strains. AB - BACKGROUND: Despite multiple causes, Chronic Kidney Disease is commonly associated with proteinuria. A previous study on Non Obese Diabetic mice (NOD), which spontaneously develop type 1 diabetes, described histological and gene expression changes incurred by diabetes in the kidney. Because proteinuria is coincident to diabetes, the effects of proteinuria are difficult to distinguish from those of other factors such as hyperglycemia. Proteinuria can nevertheless be induced in mice by peritoneal injection of Bovine Serum Albumin (BSA). To gain more information on the specific effects of proteinuria, this study addresses renal changes in diabetes resistant NOD-related mouse strains (NON and NOD.B10) that were made to develop proteinuria by BSA overload. METHODS: Proteinuria was induced by protein overload on NON and NOD.B10 mouse strains and histology and microarray technology were used to follow the kidney response. The effects of proteinuria were assessed and subsequently compared to changes that were observed in a prior study on NOD diabetic nephropathy. RESULTS: Overload treatment significantly modified the renal phenotype and out of 5760 clones screened, 21 and 7 kidney transcripts were respectively altered in the NON and NOD.B10. Upregulated transcripts encoded signal transduction genes, as well as markers for inflammation (Calmodulin kinase beta). Down-regulated transcripts included FKBP52 which was also down-regulated in diabetic NOD kidney. Comparison of transcripts altered by proteinuria to those altered by diabetes identified mannosidase 2 alpha 1 as being more specifically induced by proteinuria. CONCLUSION: By simulating a component of diabetes, and looking at the global response on mice resistant to the disease, by virtue of a small genetic difference, we were able to identify key factors in disease progression. This suggests the power of this approach in unraveling multifactorial disease processes. PMID- 16371159 TI - C/EBPbeta-2 confers EGF-independent growth and disrupts the normal acinar architecture of human mammary epithelial cells. AB - BACKGROUND: The transcription factor, C/EBPbeta, is a key regulator of growth and differentiation in the mammary gland. There are three different protein isoforms of C/EBPbeta. C/EBPbeta-1 and -2 are transactivators, and differ by only 23 N terminal amino acids present in beta-1 only. C/EBPbeta-3 (LIP) lacks the transactivation domain and represses transcription. Elevated C/EBPbeta-2 expression causes MCF10A normal human mammary epithelial cells to become transformed, undergo an epithelial to mesenchymal transition (EMT), and acquire an invasive phenotype. C/EBPbeta is a downstream transcriptional target of Ras signaling pathways and is required for Ras transformation of some cell types. Ras signaling pathways are activated in mammary epithelial cells by the ErbB receptor tyrosine kinase family. Therefore, we considered whether elevated C/EBPbeta-2 expression would resemble ErbB RTK activation in MCF10A cells. RESULTS: We show that elevated C/EBPbeta-2 expression confers EGF-independent growth in MCF10A mammary epithelial cells. However, MCF10A cells expressing C/EBPbeta-3 are not EGF-independent, and high C/EBPbeta-3 or LIP expression is incompatible with growth. C/EBPbeta-2 overexpression disrupts the normal acinar architecture of MCF10A cells in basement membrane cultures and induces complex multiacinar structures with filled lumen, similar to the consequences of aberrant ErbB2 activation. CONCLUSION: Given the ability of C/EBPbeta-2 to confer EGF independent growth to mammary epithelial cells as well as its capability for disrupting normal epithelial architecture and causing EMT, it is worth considering whether inhibitors which target ErbB family signaling pathways could be less effective in mammary epithelial cells with elevated nuclear C/EBPbeta-2 expression. PMID- 16371160 TI - Targeted infection of HIV-1 Env expressing cells by HIV(CD4/CXCR4) vectors reveals a potential new rationale for HIV-1 mediated down-modulation of CD4. AB - BACKGROUND: Efficient targeted gene transfer and cell type specific transgene expression are important for the safe and effective expression of transgenes in vivo. Enveloped viral vectors allow insertion of exogenous membrane proteins into their envelopes, which could potentially aid in the targeted transduction of specific cell types. Our goal was to specifically target cells that express the T cell tropic HIV-1 envelope protein (Env) using the highly specific interaction of Env with its cellular receptor (CD4) inserted into the envelope of an HIV-1-based viral vector. RESULTS: To generate HIV-1-based vectors carrying the CD4 molecule in their envelope, the CD4 ectodomain was fused to diverse membrane anchors and inserted together with the HIV-1 coreceptor CXCR4 into the envelopes of HIV-1 vector particles. Independent of the type of CD4 anchor, all chimeric CD4 proteins inserted into HIV-1 vector envelopes and the resultant HIV(CD4/CXCR4) particles were able to selectively confer neomycin resistance to cells expressing the fusogenic T cell tropic HIV-1 Env protein. Unexpectedly, in the absence of Env on the target cells, all vector particles carrying the CD4 ectodomain anchored in their envelope adhered to various cell types without infecting these cells. This cell adhesion was very avid. It was independent of the presence of Env on the target cell, the type of CD4 anchor or the presence of CXCR4 on the particle. In mixed cell populations with defined ratios of Env+/Env- cells, the targeted transduction of Env+ cells by HIV(CD4/CXCR4) particles was diminished in proportion to the number of Env- cells. CONCLUSION: Vector diversion caused by a strong, non-selective cell binding of CD4+-vector particles effectively prevents the targeted transduction of HIV-1 Env expressing cells in mixed cell populations. This Env-independent cell adhesion severely limits the effective use of targeted HIV(CD4/CXCR4) vectors designed to interfere with HIV-1 replication in vivo. Importantly, the existence of this newly described and remarkably strong CD4-dependent cell adhesion suggests that the multiple viral efforts to reduce CD4 cell surface expression may, in part, be to prevent cell adhesion to non target cells and thereby to increase the infectivity of viral progeny. Preventing CD4 down-modulation by HIV-1 might be an effective component of a multi-faceted antiviral strategy. PMID- 16371161 TI - Hydrogen production by Cyanobacteria. AB - The limited fossil fuel prompts the prospecting of various unconventional energy sources to take over the traditional fossil fuel energy source. In this respect the use of hydrogen gas is an attractive alternate source. Attributed by its numerous advantages including those of environmentally clean, efficiency and renew ability, hydrogen gas is considered to be one of the most desired alternate. Cyanobacteria are highly promising microorganism for hydrogen production. In comparison to the traditional ways of hydrogen production (chemical, photoelectrical), Cyanobacterial hydrogen production is commercially viable. This review highlights the basic biology of cynobacterial hydrogen production, strains involved, large-scale hydrogen production and its future prospects. While integrating the existing knowledge and technology, much future improvement and progress is to be done before hydrogen is accepted as a commercial primary energy source. PMID- 16371162 TI - Open field trial of genetically modified parthenocarpic tomato: seedlessness and fruit quality. AB - BACKGROUND: Parthenocarpic tomato lines transgenic for the DefH9-RI-iaaM gene have been cultivated under open field conditions to address some aspects of the equivalence of genetically modified (GM) fruit in comparison to controls (non GM). RESULTS: Under open field cultivation conditions, two tomato lines (UC 82) transgenic for the DefH9-RI-iaaM gene produced parthenocarpic fruits. DefH9-RI iaaM fruits were either seedless or contained very few seeds. GM fruit quality, with the exception of a higher beta-carotene level, did not show any difference, neither technological (colour, firmness, dry matter, degrees Brix, pH) nor chemical (titratable acidity, organic acids, lycopene, tomatine, total polyphenols and antioxidant capacity - TEAC), when compared to that of fruits from control line. Highly significant differences in quality traits exist between the tomato F1 commercial hybrid Allflesh and the three UC 82 genotypes tested, regardless of whether or not they are GM. Total yield per plant did not differ between GM and parental line UC 82. Fruit number was increased in GM lines, and GM fruit weight was decreased. CONCLUSION: The use in the diet of fruits from a new line or variety introduces much greater changes than the consumption of GM fruits in comparison to its genetic background. Parthenocarpic fruits, produced under open field conditions, contained 10-fold less seeds than control fruits. Thus parthenocarpy caused by DefH9-RI-iaaM gene represents also a tool for mitigating GM seeds dispersal in the environment. PMID- 16371163 TI - The Gene Set Builder: collation, curation, and distribution of sets of genes. AB - BACKGROUND: In bioinformatics and genomics, there are many applications designed to investigate the common properties for a set of genes. Often, these multi-gene analysis tools attempt to reveal sequential, functional, and expressional ties. However, while tremendous effort has been invested in developing tools that can analyze a set of genes, minimal effort has been invested in developing tools that can help researchers compile, store, and annotate gene sets in the first place. As a result, the process of making or accessing a set often involves tedious and time consuming steps such as finding identifiers for each individual gene. These steps are often repeated extensively to shift from one identifier type to another; or to recreate a published set. In this paper, we present a simple online tool which - with the help of the gene catalogs Ensembl and GeneLynx - can help researchers build and annotate sets of genes quickly and easily. DESCRIPTION: The Gene Set Builder is a database-driven, web-based tool designed to help researchers compile, store, export, and share sets of genes. This application supports the 17 eukaryotic genomes found in version 32 of the Ensembl database, which includes species from yeast to human. User-created information such as sets and customized annotations are stored to facilitate easy access. Gene sets stored in the system can be "exported" in a variety of output formats - as lists of identifiers, in tables, or as sequences. In addition, gene sets can be "shared" with specific users to facilitate collaborations or fully released to provide access to published results. The application also features a Perl API (Application Programming Interface) for direct connectivity to custom analysis tools. A downloadable Quick Reference guide and an online tutorial are available to help new users learn its functionalities. CONCLUSION: The Gene Set Builder is an Ensembl-facilitated online tool designed to help researchers compile and manage sets of genes in a user-friendly environment. The application can be accessed via http://www.cisreg.ca/gsb/. PMID- 16371164 TI - Kinetics of cancer: a method to test hypotheses of genetic causation. AB - BACKGROUND: Mouse studies have recently compared the age-onset patterns of cancer between different genotypes. Genes associated with earlier onset are tentatively assigned a causal role in carcinogenesis. These standard analyses ignore the great amount of information about kinetics contained in age-onset curves. We present a method for analyzing kinetics that measures quantitatively the causal role of candidate genes in cancer progression. We use our method to demonstrate a clear association between somatic mutation rates of different DNA mismatch repair (MMR) genotypes and the kinetics of cancer progression. METHODS: Most experimental studies report age-onset curves as the fraction diagnosed with tumors at each age for each group. We use such data to estimate smoothed survival curves, then measure incidence rates at each age by the slope of the fitted curve divided by the fraction of mice that remain undiagnosed for tumors at that age. With the estimated incidence curves, we compare between different genotypes the median age of cancer onset and the acceleration of cancer, which is the rate of increase in incidence with age. RESULTS: The direction of change in somatic mutation rate between MMR genotypes predicts the direction of change in the acceleration of cancer onset in all 7 cases (p approximately 0.008), with the same result for the association between mutation rate and the median age of onset. CONCLUSION: Many animal experiments compare qualitatively the onset curves for different genotypes. If such experiments were designed to analyze kinetics, the research could move to the next stage in which the mechanistic consequences of particular genetic pathways are related to the dynamics of carcinogenesis. The data we analyzed here were not collected to test mechanistic and quantitative hypotheses about kinetics. Even so, a simple reanalysis revealed significant insights about how DNA repair genotypes affect separately the age of onset and the acceleration of cancer. Our method of comparing genotypes provides good statistical tests even with small samples for each genotype. PMID- 16371165 TI - WebArray: an online platform for microarray data analysis. AB - BACKGROUND: Many cutting-edge microarray analysis tools and algorithms, including commonly used limma and affy packages in Bioconductor, need sophisticated knowledge of mathematics, statistics and computer skills for implementation. Commercially available software can provide a user-friendly interface at considerable cost. To facilitate the use of these tools for microarray data analysis on an open platform we developed an online microarray data analysis platform, WebArray, for bench biologists to utilize these tools to explore data from single/dual color microarray experiments. RESULTS: The currently implemented functions were based on limma and affy package from Bioconductor, the spacings LOESS histogram (SPLOSH) method, PCA-assisted normalization method and genome mapping method. WebArray incorporates these packages and provides a user-friendly interface for accessing a wide range of key functions of limma and others, such as spot quality weight, background correction, graphical plotting, normalization, linear modeling, empirical bayes statistical analysis, false discovery rate (FDR) estimation, chromosomal mapping for genome comparison. CONCLUSION: WebArray offers a convenient platform for bench biologists to access several cutting-edge microarray data analysis tools. The website is freely available at http://bioinformatics.skcc.org/webarray/. It runs on a Linux server with Apache and MySQL. PMID- 16371166 TI - Socio-demographic factors associated with smoking and smoking cessation among 426,344 pregnant women in New South Wales, Australia. AB - BACKGROUND: This study explores the socio-demographic characteristics of pregnant women who continue to smoke during the pregnancy, and identifies the characteristics of the smokers who were likely to quit smoking during the pregnancy period. METHODS: This was secondary analysis of the New South Wales (NSW) Midwives Data Collection (MDC) 1999-2003, a surveillance system covering all births in NSW public and private hospitals, as well as home births. Bivariate and multiple logistic regression analyses were performed to explore the associations between socio-demographic characteristics and smoking behaviour during pregnancy. RESULTS: Data from 426,344 pregnant women in NSW showed that 17.0% continued to smoke during pregnancy. The smoking rate was higher among teenage mothers, those with an Aboriginal (indigenous) background, and lower among more affluent and overseas-born mothers. This study also found that unbooked confinements, and lack of antenatal care in the first trimester were strongly associated with increased risk of smoking during pregnancy. About 4.0% of the smoking women reported they may quit smoking during their pregnancy. Findings showed that mothers born overseas, of higher socio-economic status, first time mothers and those who attended antenatal care early showed an increased likelihood of smoking cessation during pregnancy. Those who were heavy smokers were less likely to quit during pregnancy. CONCLUSION: Although the prevalence of smoking during pregnancy has been declining, it remains a significant public health concern. Smoking cessation programs should target the population subgroups of women at highest risk of smoking and who are least likely to quit. Effective antismoking interventions could reduce the obstetric and perinatal complications of smoking in pregnancy. PMID- 16371167 TI - Prevalence and distribution of high-risk human papilloma virus (HPV) types in invasive squamous cell carcinoma of the cervix and in normal women in Andhra Pradesh, India. AB - BACKGROUND: Despite the high incidence of cervical cancer reported from India, large scale population based studies on the HPV prevalence and genotype distribution are very few from this region. In view of the clinical trials for HPV vaccine taking place in India, it is of utmost importance to understand the prevalence of HPV genotypes in various geographical regions of India. We investigated the genotype distribution of high-risk HPV types in squamous cell carcinomas and the prevalence of high-risk HPV in cervicovaginal samples in the southern state of Andhra Pradesh (AP), India. METHODS: HPV genotyping was done in cervical cancer specimens (n = 41) obtained from women attending a regional cancer hospital in Hyderabad. HPV-DNA testing was also done in cervicovaginal samples (n = 185) collected from women enrolled in the cervical cancer screening pilot study conducted in the rural community, of Medchal Mandal, twenty kilometers away from Hyderabad. RESULTS: High-risk HPV types were found in 87.8% (n = 36/41) of the squamous cell carcinomas using a PCR-based line blot assay. Among the HPV positive cancers, the overall type distribution of the major high risk HPV types was as follows: HPV 16 (66.7%), HPV 18 (19.4%), HPV 33 (5.6%), HPV 35 (5.6%), HPV 45 (5.6%), HPV 52 (2.8%), HPV 58(2.8%), HPV 59(2.8%) and HPV 73 (2.8%). Women participating in the community screening programme provided both a self-collected vaginal swab and a clinician-collected cervical swab for HPV DNA testing. Primary screening for high risk HPV was performed using the Digene Hybrid Capture 2 (hc2) assay. All hc2 positive samples by any one method of collection were further analyzed using the Roche PCR-based line blot for genotype determination. The prevalence of high risk HPV infection in this community-based screening population was 10.3% (19/185) using the clinician-collected and 7.0% (13/185) using the self-collected samples. The overall agreement between self collected and clinician-collected samples was 92%; however among HPV-positive specimens, the HPV agreement was only moderate (39.1%). The most frequently detected HPV types in the Medchal community are HPV 52 and 16. CONCLUSION: Our results suggest that the HPV type distribution in both cervical cancer tissues and in a general screening population from Andhra Pradesh is similar to that reported in India and other parts of the world. We also conclude that an effective vaccine targeting HPV 16 will reduce the cervical cancer burden in AP. PMID- 16371168 TI - Concomitant optic nerve transection and chorioretinitis sclopetaria. AB - BACKGROUND: Optic nerve transection and chorioretinitis sclopetaria may occur following blunt ocular trauma. However, simultaneous occurrence has not yet been reported. We report the first case of concomitant optic nerve transection and chorioretinitis sclopetaria. CASE PRESENTATION: A 12-year-old boy with history of BB gun injury to his right eye was referred for loss of vision. His visual acuity was counting fingers at one meter in the right eye and with 3+ relative afferent pupillary defect (RAPD). On slit lamp examination, the right eye appeared normal except for 1+ vitreous reaction. Fundus examination of the right eye revealed a pale disc with superior retinal scar and diffuse submacular fibrosis compatible with chorioretinitis sclopetaria. Orbital CT-scans showed transection of the optic nerve by the BB gun pellet, which was lodged at the orbital apex. CONCLUSION: BB gun injury may cause concomitant optic nerve transection and chorioretinitis sclopetaria. PMID- 16371169 TI - Vestibular assessment in a patient with confirmed lateral medullary syndrome. AB - The results of vestibular investigations in a patient with confirmed lateral medullary syndrome are presented. This patient showed a unilateral weakness on caloric testing which has not been reported previously in a patient with lateral medullary syndrome. The case for the possibility of a 'central' canal paresis on caloric testing is presented. PMID- 16371170 TI - Pilonidal sinus of the neck. AB - A pilonidal sinus is most commonly seen in the sacrococcygeal region. Here we describe a rare case of pilonidal sinus of the neck following trauma due to shaving. Excision of the sinus with primary repair resulted in complete healing of the lesion with no recurrence. The possible pathogenesis of this acquired condition is also discussed. PMID- 16371171 TI - Association between indicators of cattle density and incidence of paediatric haemolytic-uraemic syndrome (HUS) in children under 15 years of age in France between 1996 and 2001: an ecological study. AB - Over the past years Shiga-like toxin-producing Escherichia coli (STEC) O157:H7 emerged as an important cause of severe gastrointestinal illnesses and haemolytic uraemic syndrome (HUS) with up to 10% of children infected with STEC developing HUS. We conducted a geographical ecological study using the district as the statistical unit. For each district, we estimated the incidence of HUS among children <15 years for the period 1996-2001 from national HUS surveillance data and data obtained on cattle density. We used multivariate Poisson regression to quantify the relation, adjusted for covariates, between paediatric HUS incidence and exposure to cattle. In univariate analysis, a positive association was observed between several cattle-density indicators and HUS incidence. In multivariate analysis, HUS paediatric incidence was associated with dairy cattle density and the ratio of calves to children <15 years (P<0.001). Our findings are consistent with previous studies in other countries and support the recommendation to limit exposure of children to dairy cattle and manure to reduce the risk of STEC infection. PMID- 16371172 TI - Hantavirus and arenavirus antibody prevalence in rodents and humans in Trentino, Northern Italy. AB - The spatial and temporal distribution of hantavirus and arenavirus antibody positive wild rodents in Trentino, Italy, was studied using immunofluorescence assays (IFA) in two long-term sites trapped in 2000-2003, and six other sites trapped in 2002. The overall hantavirus seroprevalence in the bank voles, Clethrionomys glareolus (n=229) screened for Puumala virus (PUUV) antibodies was 0.4%, and that for Apodemus flavicollis mice (n=1416) screened for Dobrava virus (DOBV) antibodies was 0.2%. Antibodies against lymphocytic choriomeningitis virus (LCMV) were found in 82 (5.6%) of the 1472 tested rodents; the seroprevalence being 6.1% in A. flavicollis (n=1181), 3.3% in C. glareolus (n=276), and 14.3% in Microtus arvalis (n=7). Of the serum samples of 488 forestry workers studied by IFA, 12 were LCMV-IgG positive (2.5%) and one DOBV-IgG positive (0.2%), however, the latter could not be confirmed DOBV-specific with a neutralization assay. Our results show a widespread distribution but low prevalence of DOBV in Trentino, and demonstrate that the arenavirus antibodies are a common finding in several other rodent species besides the house mouse. PMID- 16371173 TI - Impact of water supply, domiciliary water reservoirs and sewage on faeco-orally transmitted parasitic diseases in children residing in poor areas in Juiz de Fora, Brazil. AB - The objectives of this study were to characterize faeco-orally transmitted parasitic diseases and to identify the factors associated with these diseases, with emphasis on environmental factors, in children ranging from 1 up to 5 years old residing in substandard settlement areas. A population-based cross-sectional epidemiological design was used in a non-random selection of 29 out of the 78 substandard settlement areas in the municipality of Juiz de Fora, Brazil. A sample of 753 children were assessed from the target population consisting of all children of the appropriate age range residing in the selected areas. Data were collected by means of domiciliary interviews with their mothers or with the person responsible for them. The Hoffmann-Pons-Janer method was used in the parasitological examination of faeces. Binary logistic regression models were used to identify the factors associated with the diseases. A total of 319 sample children presented faeco-orally transmitted parasitic diseases. The factors associated with these parasitic diseases included the children's age, family income, number of dwellers in the domicile, consumption of water from shallow wells, consumption of water from natural sources, absence of covered domiciliary water reservoirs, and the presence of sewage flowing in the street. PMID- 16371174 TI - The SARS outbreak in a general hospital in Tianjin, China -- the case of super spreader. AB - Severe acute respiratory syndrome (SARS) is a newly emerged infectious disease with a high case-fatality rate and devastating socio-economic impact. In this report we summarized the results from an epidemiological investigation of a SARS outbreak in a hospital in Tianjin, between April and May 2003. We collected epidemiological and clinical data on 111 suspect and probable cases of SARS associated with the outbreak. Transmission chain and outbreak clusters were investigated. The outbreak was single sourced and had eight clusters. All SARS cases in the hospital were traced to a single patient who directly infected 33 people. The patients ranged from 16 to 82 years of age (mean age 38.5 years); 38.7% were men. The overall case fatality in the SARS outbreak was 11.7% (13/111). The outbreak lasted around 4 weeks after the index case was identified. SARS is a highly contagious condition associated with substantial case fatality; an outbreak can result from one patient in a relatively short period. However, stringent public health measures seemed to be effective in breaking the disease transmission chain. PMID- 16371175 TI - Sorbitol-fermenting Shiga toxin-producing Escherichia coli O157: indications for an animal reservoir. AB - This study investigates a sorbitol-fermenting enterohaemorrhagic Escherichia coli (SF EHEC) O157 infection in a farmer's family in the Austrian province of Salzburg. The investigation commenced after a 10-month-old boy was admitted to hospital with the clinical diagnosis of a haemolytic-uraemic syndrome (HUS) and his stool specimen grew SF EHEC O157:H-. In a subsequent environmental survey, a stool specimen of the 2-year-old brother and faecal samples of two cattle from the family's farm were also found to be positive for SF EHEC O157:H-. All four isolates had indistinguishable phenotypic and molecular characteristics and were identical to the first strain detected in Bavaria in 1988. Despite identical isolates being demonstrated in Bavaria after 1988, and until this report, increased surveillance in neighbouring Austria had not found this organism. We propose that the strain may have recently spread from Bavaria to Austria. Although SF EHEC O157:H- strains are still rare, they may represent a considerable health threat as they can spread from farm animals to humans and between humans. PMID- 16371176 TI - Haemolytic uraemic syndrome associated with interfamilial spread of E. coli O26:H11. AB - In September 2000, haemolytic uraemic syndrome (HUS) was diagnosed in a 10-month old child with a prodromal history of vomiting and diarrhoea (non-bloody). Investigation revealed that a self-limiting gastrointestinal illness (mean duration 48 h) had occurred among immediate and extended family in the 2 weeks prior to the child's admission. The epidemiology of the illness suggested person to-person spread. Five children (close family contacts) had E. coli O26 verocytotoxin (VT1 and VT2) isolated from stools. Stool culture and serology from the index case were negative for shiga toxin-producing E. coli (STEC) organisms. Control measures in accordance with the Public Health Laboratory Service (PHLS), verocytotoxogenic organisms (VTEC) guidelines were applied to prevent further spread among the extended family and contacts. Despite detailed food and environmental exposure histories, the source of the illness was not identified. This incident highlights the importance of investigation of cases of post diarrhoeal HUS, for potential shiga toxin E. coli aetiology. PMID- 16371177 TI - Listeria spp. in the coastal environment of the Aqaba Gulf, Suez Gulf and the Red Sea. AB - Listeria monocytogenes is an important pathogen which causes an infection called listeriosis. Because of the high mortality rate (~30%) associated with listeriosis, and the widespread nature of the organism, it is a major concern for food and water microbiologists since it has been isolated from various types of foods, including seafood, as well as from the aqueous environment. To investigate the prevalence of this pathogen in the Aqaba Gulf (12 sites), Suez Gulf (14 sites) and Red Sea (14 sites), 200 water samples (collected during five sampling cruises in 2004), 40 fresh fish samples and 15 shellfish samples were analysed using the enrichment procedure and selective agar medium. All water samples were also examined for the presence Listeria innocua which was the most common of the Listeria spp. isolated, followed by L. monocytogenes, with a low incidence of the other species. During the whole year, the percentage of Listeria spp. and L. monocytogenes in 200 water samples was 20.5% (41 samples) and 13% (26 samples) respectively. In fresh fish (40 samples) it was 37% (15 samples) and 17.3% (7 samples) and in shellfish (15 samples) 53% (8 samples) and 33% (5 samples) respectively. In water samples, there was an association between the faecal contamination parameters and the presence of the pathogen; however, water salinity, temperature, dissolved oxygen and pH did not influence the occurrence of this bacterium. These results may help in the water-quality evaluation of the coastal environments of these regions. PMID- 16371178 TI - The distribution of travel-associated Legionnaires' disease within selected European countries, and a comparison with tourist patterns. AB - Much data has been gathered by the EWGLINET scheme on the distribution of cases of travel-associated Legionnaires' disease (TALD) by country of infection, but less analysis has been carried out on the distribution of these cases within countries. Travel-associated cases with onset in 2002 linked to France, Italy, Spain and Turkey were mapped. Rates of Legionnaires' disease per 100,000 tourists were calculated for internal and foreign visitors for the regions of each country, and mapped. Rates of 1.5 cases/100,000 and 2 cases/100,000 tourists were classified as 'high' and 'very high' respectively. Cases of TALD were concentrated in certain regions, but when rates were calculated using tourist data, the results were relatively constant throughout each country. Rates were higher among foreign visitors than internal visitors; three of the countries had at least one region with 'high' rates, whilst Turkey additionally had three regions with 'very high' rates. PMID- 16371179 TI - Vaccination coverage in adults undergoing splenectomy: evaluation of hospital vaccination policies. AB - Vaccination coverage in 595 adult patients undergoing total splenectomy in the Hospital Clinic of Barcelona during 1992-2002 was studied. The rates of cover for pneumococcal, Haemophilus influenzae type b and meningococcal vaccines were 63, 63 and 61% respectively, during 2000-2002; 32, 17 and 22% in 1997-1999; and 24, 9 and 8% in 1992-1996. Multivariate analysis showed a greater risk of no vaccination in splenectomies due to trauma, malignant neoplasms of solid organs and incidental splenectomy compared with both neoplastic and non-neoplastic haematological disease, and those patients undergoing splenectomy before 2001. Coverage (>/=1 vaccine) since 1997 in patients with haematological diseases was 83.5% (71/85), haematological neoplasias 69.2% (18/26), solid organ neoplasms 38.3% (36/94), incidental splenectomy 35.6% (16/45), and traumas 28.4% (21/74). Mandatory hospital admission of patients undergoing splenectomy offers a good opportunity for vaccination of these patients. Specific vaccination policies should be developed to take advantage of this circumstance. PMID- 16371180 TI - Specific IgM and IgG responses in primary and secondary dengue virus infections determined by enzyme-linked immunosorbent assay. AB - IgM- and IgG-capture ELISAs are widely used as diagnostic tests for confirmation of dengue virus infection. The positive rate of anti-dengue IgM and IgG detection was examined in primary and secondary dengue virus infections in the setting of a provincial hospital using IgM- and IgG-capture ELISAs. Disease day 1 was defined as the day of onset of symptoms. In total, 232 plasma samples were collected from 106 confirmed dengue cases consisting of 12 primary and 94 secondary infections. In primary infection, anti-dengue IgM was detected in 4 out of 5 samples collected on disease day 5 and in all the 21 samples collected on disease day 6 or later. Specific IgG was detected in 2 out of 5 samples collected on day 12, and in 5 out of 6 samples collected on disease days 13-15, but was not detected in samples collected on disease day 10 or earlier. In secondary infection, IgM was not detected in the samples on disease days 2 and 3, but detected in 20 out of 79 samples collected on days 4-6, in 44 out of 65 on disease days 7-11 and in 40 out of 51 samples on disease days 12-14. In contrast, specific IgG was detected in 21 out of 60 samples on disease days 4 and 5, in 13 out of 19 on disease day 6, in 62 out of 65 on disease days 7-11 and in all the samples collected on disease day 12 or later. The result indicate that seroconversion rates of IgM and IgG are different between primary and secondary infections, and suggest that detection of specific IgM and IgG is necessary for determining dengue virus infection and for differentiating primary and secondary dengue infections. PMID- 16371181 TI - An evaluation and comparison of three commonly used statistical models for automatic detection of outbreaks in epidemiological data of communicable diseases. AB - We evaluated three established statistical models for automated 'early warnings' of disease outbreaks; counted data Poisson CuSums (used in New Zealand), the England and Wales model (used in England and Wales) and SPOTv2 (used in Australia). In the evaluation we used national Swedish notification data from 1992 to 2003 on campylobacteriosis, hepatitis A and tularemia. The average sensitivity and positive predictive value for CuSums were 71 and 53%, for the England and Wales model 87 and 82% and for SPOTv2 95 and 49% respectively. The England and Wales model and the SPOTv2 model were superior to CuSums in our setting. Although, it was more difficult to rank the former two, we recommend the SPOTv2 model over the England and Wales model, mainly because of a better sensitivity. However, the impact of previous outbreaks on baseline levels was less in the England and Wales model. The CuSums model did not adjust for previous outbreaks. PMID- 16371182 TI - Isolation of Vibrio parahaemolyticus and Vibrio vulnificus from wild aquatic birds in Japan. AB - Vibrio parahaemolyticus and Vibrio vulnificus were isolated from faecal samples of wild aquatic birds in winter. Although V. parahaemolyticus and V. vulnificus were present in low numbers in seawater in the area where the faecal samples of the birds were collected, the pathogens were isolated from the faeces of the birds. This study demonstrates that wild aquatic birds are a vehicle for V. parahaemolyticus and V. vulnificus to survive in winter. PMID- 16371183 TI - A syringe exchange programme in prison as prevention strategy against HIV infection and hepatitis B and C in Berlin, Germany. AB - In two prisons in Berlin, Germany, provision of sterile injection equipment for injecting drug users (IDUs) started in 1998. To assess the programme's impact, the frequency of injecting drug use and syringe sharing, and the incidence of HIV, HBV, and HCV infection were determined in a follow-up study. Of all IDUs (n=174), 75% continued to inject. After the project start the level of syringe sharing declined from 71% during a 4-month period of previous imprisonment to 11% during the first 4 months of follow-up, and to virtually zero thereafter. Baseline seroprevalences for HIV, HBV, and HCV were 18, 53, and 82%. HIV and HCV seroprevalence at baseline was significantly associated with drug injection in prison prior to the project start. No HIV and HBV seroconversions, but four HCV seroconversions occurred. The provision of syringes for IDUs in appropriate prison settings may contribute to a substantial reduction of syringe sharing. However, the prevention of HCV infection requires additional strategies. PMID- 16371184 TI - [Extension of lymphadenectomy in colorectal surgery: open subject]. PMID- 16371185 TI - Which therapy to prevent post-thyroidectomy hypocalcemia? AB - Hypocalcemia is one of the most frequent complications after total extracapsular thyroidectomy (TET). In most of cases it is a transient phenomenon. The aim of this study is to evaluate if and how the oral administration of calcium or calcium combined with D-vitamin could effectively prevent post-thyroidectomy hypocalcemia. A randomized prospective study was performed, recruiting 120 patients who underwent total thyroidectomy. The patients in our series were randomly assigned to one of two groups: group A--patients who received calcium lactogluconate/calcium carbonate (mg 300 per day); group B--patients who received calcium carbonate/cholecalciferol therapy (calcium carbonate: 1500 mg per day; cholecalciferol 400 UI per day). The groups were well matched for age, sex and pathologies. Patients of both A and B groups were divided in two subgroups: those operated on for benign thyroid diseases (A1 and B1) and those operated on for malignancy (A2, B2). Serum calcium assays, performed 24, 48 and 72 hours after surgery, showed mean values of calcemia higher in patients of the B1 and B2 group. Statistical analysis was performed using a Student's t test. Mean serum calcium concentrations on post-operative day one, two and three were higher in patients of the group B (p<<0.01). Early and combined oral administration of both calcium and vitamin D seemed to prove major efficacy in preventing and treating post-operative hypocalcemia, showing mean serum calcium levels higher than those of patients who received only oral calcium administration. Nevertheless, further studies are necessary to validate these data. PMID- 16371186 TI - Intra-abdominal inflammatory myofibroblastic pseudotumor: case report and review of the literature. AB - Inflammatory myofibroblastic pseudotumors (IPM) are very rare tumor characterized by unpredictable clinical behaviour. They arise in soft tissues of almost every organ and the most common site is the lung. Over 200 cases of inflammatory myofibroblastic pseudotumor of the lung have been described in literature. Intra abdominal IMP are very rare. We describe a case of intra-abdominal IMP in a boy of 15 years old who presented symptoms and signs of acute appendicitis. Exploratory laparotomy revealed a mass in the peritoneal cavity. The mass was removed. The histologic examination showed that it was an IMP. Surgical treatment was the only therapy. Six months after the surgical operation the patient has no sign of illness. PMID- 16371187 TI - [Colorectal carcinoma in the young. Prognostic factors]. AB - Colorectal carcinoma is the third most frequently diagnosed malignant neoplasm. Usually patients affected by this neoplasia belong to VI decade of life. However approximately 2-8% of tumors arise in patients with age under 40 years. Aim of the study was to analyse the results of surgical treatment of colorectal cancer in patients aged under forty. From January 1987 to December 2002, 46 patients under forty years with colorectal cancer underwent surgical procedure. No perioperative mortality was registered, and complications were evidenced in nine patients (20%). Actuarial five years survival was 33%, and overall mean survival was 53 months. Univariate and multivariate analyses identified as prognostic factors the tumor grade, Dukes' stage, nodal status, and length of symptoms. PMID- 16371188 TI - [A serious complication in colorectal surgery: anastomotic leakage. Our experience]. AB - Anastomotic leakage is a severe complication in colorectal surgery with a lot of generic and specific risk factors. There are still controversies about the possibility to prevent it or to limit its severity with the use of faecal diversions. We report our experience on 189 patients operated in five years on colon-rectum, in election and emergency, with manual or mechanical anastomoses in the last 20 cm from the anal verge. We have had 21 anastomotic leakages (11%), symptomatic in 10 cases (5.5%), and 5 deaths (2.6%), 3 directly correlated to the leakage. In the patients (n 98) with loop-ileostomy there wasn't anastomotic leakages reduction, but only of clinical impact. We prefer loop-ileostomy in cases of extraperitoneal anastomoses, always in emergency and in anastomoses at risk. PMID- 16371189 TI - [Anisakiasis presenting as perforated ulcer of the cecum]. AB - A case of cecal anisakiasis is presented. Symptoms were compatible with appendicitis. The patient was operated and a perforated ulcer of the cecum was found. Ileocolic resection was performed. The histologic result showed the presence of Anisakis simplex larva in the muscle of the cecum. The patient was discharged the fifth day without complications. At present he is asymptomatic. He had eaten uncooked anchovies some days before the onset of the disease. PMID- 16371190 TI - [Observations about a case of giant splenic cyst]. AB - The Authors, on the basis of a case of giant spleen cyst with positive tumoral markers, analyse some epidemiological and clinical aspects related to splenic non parasitic cysts. They affirm the priority of the conservative surgery, whenever possible, followed by an appropriate follow-up, although in this case their therapeutic choice was radical, due to the lack of residual parenchyma. In accordance with the data of several publications, as well as on the basis of the results obtained, the conservative approaches have been reevaluated, above all in view of the modern findings related to the function of the spleen. The conservative approach cannot be carried out in the following cases: neoplastic diseases, increase of the tumoral markers serum levels, total involvement of the splenic parenchyma by cysts. PMID- 16371191 TI - [Gangrene of Meckel's diverticulum in strangulated left inguinal hernia]. AB - We report a case of a 57-year-old woman admitted for abdominal pain and a not reducible mass in left inguino-abdominal region. With a diagnosis of strangulated inguinal hernia, the patient underwent urgent surgery. The surgical exploration showed a gangrenous intestinal loop with a Meckel's necrotic diverticulum. A small bowel resection (20 cm) was performed. The post-operative course was uneventful. This seems the first case reported in the literature of woman with a Meckel's diverticulum involved in a strangulated left inguinal hernia. PMID- 16371192 TI - Major complications in thyroid surgery: utility of bipolar vessel sealing (Ligasure Precise). AB - In the present study the Authors tried and assess the advantages of the standard sutureless thyroidectomy performed by the Ligasure Vessel Sealing System, thanks to the use of the dedicated Precise handle. The Authors compared the efficacy of haemostasis and the economical impact of the device, in terms of drug administration and costs. The Authors comparatively analyzed 120 total extracapsular thyroidectomies (TET) performed by the standard operative technique (Group A, control) and 70 TET achieved by the "sutureless technique" (Group B, case). There was a statistically significant decrease of transient postoperative hypocalcemia (5.71% vs 7.5%) and also of mean operative time (about 20 minutes) in patients of the group B. Non significant decrease of other kind of complications (postoperative hemorrhage, transient and permanent inferior laryngeal palsy, stupor of the superior laryngeal nerve, seromas) were also observed. The use of the Ligasure Precise resulted easy, safe and efficient in the Authors' experience. It allowed the decrease of postoperative haemorrhages and mean operative time. PMID- 16371193 TI - [Thromboembolic risk and prevention of deep venous thrombosis in open and laparoscopic surgery]. AB - Deep Vein Thrombosis (DVT) and pulmonary embolism are the dangerous and serious complications in patients undergoing surgery. It is known that prognosis is strictly linked to timely recognition of the pathogenetic-clinical phase of the thromboembolic disease and that prevention, therefore, plays the leading role in patients at risk. The most recent series show that, in absence of prophylaxis, the frequency of DVT, diagnosed by objective tests, is still significant in abdominal surgery. Modern diagnostic tools make possible to identify relatively silent clinical thrombosis, also with laboratory tests (i.e., D-dimer plasma levels). The Authors report a study on thromboembolic episodes in patients who underwent pneumoperitoneum with CO2 during laparoscopic abdominal surgery, compared to a control group submitted to open surgery. They underline the importance of a careful preoperative evaluation of the venous system, by Doppler study, in order to identify, patients at risk of DVT and establish a suitable anti-thrombotic prophylaxis. PMID- 16371194 TI - [A new international diabetes federation worldwide definition of the metabolic syndrome: the rationale and the results]. PMID- 16371195 TI - [Use of cost-effectiveness analysis to guide the clinical implementation of new therapies]. PMID- 16371196 TI - [Congenital heart disease: present situation and future prospects]. PMID- 16371197 TI - [Short-and long-term cost-effectiveness analysis of adding clopidogrel to standard therapy in acute coronary syndrome patients in Spain]. AB - INTRODUCTION AND OBJECTIVES: The CURE study showed that adding clopidogrel to standard therapy with acetylsalicylic acid reduces the risk of cardiovascular events (i.e., stroke, myocardial infarction, and cardiovascular death) in patients with acute coronary syndrome but without ST-segment elevation. The objective of this study was to carry out short- and long-term cost-effectiveness analyses of administering clopidogrel in addition to standard therapy during the first year of treatment. PATIENTS AND METHOD: For the short-term analysis, clinical data and information on health resource utilization were taken from the CURE study. For the long-term analysis, an adaptation of an internationally used Markov model involving six health states was employed. Clinical data were obtained from clinical trials and epidemiological studies. Information on resource use was obtained from two Spanish registries of patients with acute coronary syndrome, a literature review, and consultations with an expert panel. Results are expressed in terms of incremental cost per event avoided or per life year gained. RESULTS: In the short-term analysis, the incremental cost per event avoided of adding clopidogrel to standard therapy was ;17 190. In the long-term analysis, the incremental cost per life-year gained was ;8132, which is below the Spanish cost-effectiveness threshold of ;30 000 per life-year gained. CONCLUSIONS: Adding clopidogrel to standard therapy during the first year of treatment is cost-effective in both the short and long term. PMID- 16371198 TI - [Effect of reactions to symptoms onset on early mortality from myocardial infarction]. AB - INTRODUCTION AND OBJECTIVES: A patient's social circumstances at the time when acute myocardial infarction (AMI) symptoms first appear might influence survival. Our objectives were to study the living conditions, the location where symptoms started, the type of symptoms, and the delay before action was taken in patients with AMI who survived more than one hour, and to analyze the relationship between these variables and mortality in different time periods. PATIENTS AND METHOD: Population-based observational cohort study carried out in 1997-1998. Main data source: Registre Gironi del Cor (REGICOR). Death certificates provided information on patients who died before they could be included in the register. The patients' demographic characteristics, lifestyle, clinical history, electrocardiographic abnormalities, cardiac enzyme levels, treatment, and diagnosis were recorded. Mortality before and during hospitalization, and overall mortality at 28 days were studied. RESULTS: Of the 1,097 patients included, 274 (24.97%) died before reaching hospital, 171 (15.58%) died in hospital, and 652 (59.4%) were alive at 28 days. Mortality was lower in patients who went directly to hospital (OR = 0.32, 95% CI, 0.17-0.59). Mortality at 28 days was higher in those with atypical symptoms (OR = 5.52, 95% CI, 2.90-10.50), and in those who lived in an institution (OR = 9.47, 95% CI, 1.05-84.9). CONCLUSIONS: In the absence of specially equipped ambulances, AMI patients who went directly to the hospital or who had typical symptoms had a better chance of survival both before hospitalization and at 28 days. In contrast, 28-day mortality was higher in institutionalized patients. PMID- 16371199 TI - [Underrecognized peripheral arterial disease in patients with acute coronary syndrome: prevalence of traditional and emergent cardiovascular risk factors]. AB - INTRODUCTION AND OBJECTIVES: Peripheral arterial disease (PAD) frequently coexists with coronary artery disease. Our objective was to determine the prevalence of traditional and emergent cardiovascular risk factors in patients with acute coronary syndrome (ACS), with or without PAD. PATIENTS AND METHOD: A prospective study of 141 consecutive patients (< 70 years old) admitted to our hospital with ACS was performed. PAD was diagnosed when the ankle-brachial index (ABI) was < or = 0.9. Traditional cardiovascular risk factors were evaluated. C reactive protein, homocysteine, amyloid A, lipoprotein (a), fibrinogen, apolipoprotein A1, and apolipoprotein B100 serum levels, and microalbuminuria were measured. Specific genotypes were also determined. RESULTS: Patients were divided into two groups according to whether PAD was present (37 patients, 26% of total, ACS-PAD group) or absent (104 patients, ACS group). In the ACS-PAD group, patients were older, and diabetes and hypertension were significantly more common. Moreover, levels of C-reactive protein (3.1 mg/L vs 2.18 mg/L; P<.05), homocysteine (11.45 mmol/L vs 9.4 mmol/L; P<.01), amyloid A (5.2 mg/mL vs 3.7 mg/mL; P<.05), and microalbuminuria (4.89 mg/L vs 3.1 mg/L; P<.05) were significantly higher in this group. Logistic regression analysis showed that poorly controlled diabetes (OR = 6.3; 95% CI, 1.1-36.7), time-dependent tobacco exposure (OR = 1.5 per decade; 95% CI, 1.2-2.0), and high pulse pressure (OR = 1.9 per 10 mmHg; 95% CI, 1.3-2.7) were independent predictors of the presence of PAD. CONCLUSIONS: Several traditional and emergent cardiovascular risk factors were more prevalent in patients with acute coronary syndrome and peripheral arterial disease. Moreover, some factors were independent predictors of peripheral arterial disease. PMID- 16371200 TI - [Prevalence of cardiovascular risk factors in the Balearic Islands (CORSAIB Study)]. AB - INTRODUCTION AND OBJECTIVES: Indices of cardiovascular morbidity and mortality in the Balearic Islands in Spain are poor. We decided to investigate the prevalence of the main cardiovascular risk factors (CRFs) and of different cardiovascular diseases in inhabitants of these islands aged 35-74 years. SUBJETS AND METHOD: A population-based descriptive cross-sectional study was used. The Balearic Islands were divided into 14 sectors, each with three or four participating general practitioners (GPs). A random selection of 40 patients registered with each GP was made. Patients were contacted in the surgery or by mail or telephone. Each GP administered a questionnaire on the patients' family and personal history of cardiovascular disease and CRFs such as levels of smoking and physical activity, carried out anthropometric and blood pressure measurements, and requested blood glucose, cholesterol, and cholesterol fraction measurements. RESULTS: The study included 1685 individuals. Some 27% smoked (men 36.9%, women 18.7%), 47.8% had high blood pressure (men 52.3%, women 43.4%), 24.2% had hypercholesterolemia (men 24.4%, women 24.1%), 11.7% had diabetes mellitus (men 15.3%, women 8.4%), 27% were obese (men 24.8%, women 29%), 40.1% were overweight (men 48.3%, women 33.4%), and 43.1% of men and 45.6% of women were sedentary. Two or more CRFs were observed in 58.4%, and 9.8% had a diagnosed cardiovascular disease. The Framingham method gave a higher estimate of cardiovascular risk in all age groups than the REGICOR method. CONCLUSIONS: The prevalence of the main CRFs in the Balearic Islands is high, implying a high risk profile. Dietary and lifestyle interventions, and preventive measures are recommended. PMID- 16371201 TI - [Relationship between coronary flow and viability in patients with myocardial infarction submitted to revascularization with stent. Assessment by Doppler guidewire]. AB - Introduction and objectives. Intracoronary Doppler guidewire measurement provides information about the state of the myocardial microcirculation. Our aim was to assess the relationship between the pattern of coronary flow and myocardial viability. Patients and method. The study included 19 patients who had had a recent myocardial infarction (< 1 month), who underwent percutaneous revascularization of the responsible vessel, and who agreed to angiographic follow-up. Patients were divided into two groups according to the extent of the akinetic area observed at follow-up responsible (149 [40] days) in the myocardial region supplied by the responsible artery: one group (n=11) had preserved regional contractility (i.e., an akinetic area less than 30%); the other group (n=8) had impaired regional contractility (i.e., an akinetic area equal to or greater than 30%). Coronary flow in the artery was responsible assessed after revascularization and at follow-up using the Doppler guidewire method. RESULTS: After revascularization, patients with preserved regional contractility had a higher diastolic deceleration half-time than those with impaired regional contractility (628 [188] ms vs 376 [131] ms, P=.02). At follow-up, the difference in coronary flow between the groups had disappeared. Patients in whom regional contractility was preserved demonstrated a significant increase in coronary flow reserve at follow-up (1,8 [0,5] vs 2,3 [0,5], P=.01). CONCLUSIONS: After timely revascularization (within one month) of the vessel responsible for a myocardial infarction, the pattern of coronary flow observed in patients with viable myocardium differed from that in those with non-viable myocardium. The differences disappeared during follow-up. PMID- 16371202 TI - [Incidence of congenital heart disease in navarra (1989-1998)]. AB - INTRODUCTION AND OBJECTIVES: Congenital cardiopathies are the most common forms of congenital malformation. They occur in between 5.2 and 12.5 in every thousand live births. The aim of this study was to describe the incidence and nature of congenital heart disease in the Spanish region of Navarra during a specified time period (1989-1998). PATIENTS AND METHOD: The study involved all children with congenital heart disease among the 47 783 born in the region in the specified time period. RESULTS: The incidence was 8.96 per thousand live births, with 90% having one of the 10 most common types of cardiac malformation. The accumulative percentage diagnosed was 25.3% in the first 24 hours of life, 45% in the first week, 65% in the first month, and 83.1% during the first year. Some 30.8% of cases of congenital heart disease required invasive treatment: 25.4% underwent surgery and 6.4%, cardiac catheterization. CONCLUSIONS: The incidence of congenital heart disease in Navarra falls within the range reported for developed countries. The level of care provided in this region is good, as demonstrated by existing diagnostic capabilities and treatment provision. PMID- 16371203 TI - [Spanish implantable cardioverter-defibrillator registry. First official report of the spanish society of cardiology working group on implantable cardioverter defibrillators (2002-2004)]. AB - OBJECTIVE: To report the 2002-2004 findings of the Spanish National Implantable Cardioverter-Defibrillator (ICD) Registry, established by the Spanish Society of Cardiology Working Group on Implantable Cardioverter-Defibrillators. MATERIAL AND METHOD: Data were collected prospectively after implantation using a single-page questionnaire returned to the Spanish Society of Cardiology. Participation was voluntary. RESULTS: The registry received reports on 925, 1,046 and 1414 implants, respectively, in the years 2002, 2003 and 2004. These figures represent 63%, 59% and 67.5%, respectively, of the total number of ICDs implanted. The reported implantation rates were 22, 24 and 33 per million, respectively, and the estimated total implantation rates were 35, 41 and 49, per million, respectively. The number of device replacements increased from 20% to 30% between 2002 and 2004. The majority of patients were male, their median age was 66 years, they had severe or moderate left ventricular dysfunction, and they were in functional class I or II. The most common underlying heart disease was ischemic heart disease. The main indications for an ICD were sustained monomorphic ventricular tachycardia and aborted sudden cardiac death, though the number of prophylactic indications has increased. Most ICDs were implanted in an electrophysiology laboratory by a cardiac electrophysiologist. The implantation rates of dual chamber ICDs and ICDs with cardiac resynchronization therapy were approximately 30% and 15%, respectively. Very few complications occurred during implantation. CONCLUSIONS: The Spanish National ICD Registry contains a representative sample of ICD implantations performed in the country. The registry is one of the largest reported. PMID- 16371204 TI - [Spanish catheter ablation registry. Fourth official report of the spanish society of cardiology working group on electrophysiology and arrhythmias (2004)]. AB - INTRODUCTION: This article details the findings (i.e., success and complication rates) of the 2004 Spanish Catheter Ablation Registry, which was established by the Spanish Society of Cardiology Working Group on Electrophysiology and Arrhythmias. This is the fourth consecutive annual report. MATERIAL AND METHOD: Similar to last year, data were collected in two ways: retrospectively using a standard questionnaire, and prospectively using a central database containing details of each ablation procedure. Each center was asked to participate by selecting one of these methods. RESULTS: Thirty-six centers contributed voluntarily to the registry. Overall, 4147 ablation procedures were reported, giving a mean of 115 [66] procedures per center. The three main conditions treated were AV node reentry tachycardia (32%), the presence of accessory pathways (25%), and common atrial flutter (23%). Success rates were 98%, 89% and 91%, respectively. The overall success rate was 90%, with a major complication rate of 1.4% and a mortality rate of 0.07%. CONCLUSIONS: Although the participation rate was slightly lower than in previous years, the comparability of data collected on a substantial number of ablation procedures (more than 4000) with earlier registry data confirm the validity and consistency of the register. PMID- 16371205 TI - [New prospects in myocardial surgical revascularization]. AB - Myocardial revascularization with coronary bypass graft surgery or angioplasty is the conventional treatment for ischaemic heart disease. The progressive refinement of both techniques (eg, use of arterial grafts and drug eluting stents) has improved clinical outcomes but the elucidation of which of the two treatments is more effective would require well designed randomised studies. While the utility of off-pump as compared to on-pump coronary bypass graft surgery is still debated, the evidence for the exclusive use of arterial conduits for surgical revascularization is compelling. Patients with diffuse coronary artery disease that cannot be treated by conventional revascularization treatments remain a challenge for the control of angina and a variety of alternative treatments, including transmyocardial laser revascularization and cardiac denervation, have been applied. However, these treatments do not eliminate angina and the benefit obtained is transient. The angiogenic therapy using stem cells and modification of gene expression is a new treatment which potential, in spite of the promising animal studies and the initial clinical trials, is still unclear. The introduction of less invasive surgical techniques and robotics could play an important role for the delivery of these treatments in the future. PMID- 16371206 TI - [Optical coherence tomography imaging in coronary drug-eluting stenting]. PMID- 16371207 TI - [Unusual cyanosing heart defect due to supravalvular tricuspid obstruction in an infant]. AB - We present a case involving a rare form of cyanotic congenital heart disease in a 2-month-old neonate. The initial diagnosis was thought to be tricuspid dysplasia with right-to-left shunting through an interatrial communication. However, surgery showed the presence of a pedunculated mass that prolapsed into the tricuspid valve orifice, thereby causing severe obstruction to right ventricular filling. This mass was attached to the lower part of the atrial septum. Pathological examination showed fibrotic myxoid tissue. After the mass was resected surgically, the tricuspid valve appeared normal. We comment on the pathogenesis and the differential diagnosis of this rare entity. PMID- 16371208 TI - [Percutaneous closure of pseudoaneurysm of the mitral-aortic intervalvular fibrosa]. AB - Pseudoaneurysm of the mitral-aortic intervalvular fibrosa is an uncommon event, which is usually secondary to endocarditis of the aortic valve. Its clinical evolution is variable and potentially serious complications can occur. Therefore, surgical treatment is usually recommended. To the best of our knowledge, this is the first description of percutaneous treatment of this disease. PMID- 16371209 TI - [Cardiac cephalgia: an underdiagnosed condition? ]. AB - Cardiac cephalgia, or headache occurring as manifestation of myocardial ischemia, has only recently been recognized as a distinct entity. In patients with known ischemic cardiopathy, its diagnosis depends on the presence of severe headache that is accompanied by nausea, worsened by physical exercise, and only ceases with nitrate administration. We report on two patients who met diagnostic criteria for this entity. In both, headache was the only symptom of coronary ischemia, and delayed its diagnosis. Headache occurred both at rest and during exertion, and resolved only after the administration of nitrates. Cardiac cephalgia should be suspected in patients with a history of ischemic cardiopathy who present with de novo headache, even when thoracic pain is absent, especially if the headache improves with nitrates. Differential diagnosis with migraine is crucial to avoid the administration of vasoconstrictors. PMID- 16371210 TI - [Percutaneous intervention for severe aortic stenosis and left main artery disease in a morbidly obese patient]. AB - Obesity increases significantly the rate of postsurgical complications and mortality in patients undergoing major surgery. We present the case of a morbidly obese 65-year-old female with severe aortic stenosis and left main coronary artery disease who underwent successful aortic valvuloplasty and angioplasty, with placement of a stent in the left main coronary artery. After undergoing bariatric surgery and losing 30% of her body weight, the patient was accepted for cardiac surgery to replace the aortic valve and to bypass the left anterior descending coronary artery using the mammary artery. There were no surgical complications. PMID- 16371211 TI - [Revista Espanola de cardiologia 2005: activity and scientific recognition]. PMID- 16371212 TI - [Do we need a randomized trial (TRIANA) on reperfusion in patients aged over 75 years?]. PMID- 16371214 TI - Wheeze: the first episode. PMID- 16371215 TI - Health-economic analyses of subcutaneous specific immunotherapy for grass pollen and mite allergy. AB - OBJECTIVE: To investigate the health and monetary consequences of treating allergy with specific immunotherapy (SIT) compared with symptomatic treatment/standard care among patients with grass pollen or mite allergy. METHODS: We performed an economic analysis based on 253 grass- and/or mite allergic patients who started SIT from 1.1.1996 to 1.1.2002 at the Allergy Unit, Aarhus University Hospital and at a specialist practice in Aarhus. Relevant data were collected before, during and after SIT treatment from the national health service based on each patient's personal identification number and medical records and from a specifically designed questionnaire. A cost-benefit analysis including direct and indirect costs before, during and after SIT was performed. In addition direct costs were related to the clinical effect (improvement in well being) in the form of a cost-effectiveness analysis. RESULTS: The direct cost per patient/year before SIT (equivalent to standard care) was DKK 2,580. The investment in SIT was DKK 27,545 (in present values) per patient over a 4-year period. After SIT the cost was reduced to DKK 1,072 per patient/year. In the long term, prospective introduction of SIT incurred additional present-value direct costs of DKK 13,676 per patient treated and DKK 2,784 per patient/year of improved well-being. However, when indirect costs were included in the economic evaluation SIT was shown to be net beneficial. CONCLUSION: This study reveals that SIT is associated with initial resource investments and subsequent resource savings in the long term compared with standard care. When all consequences are measured in monetary terms, and assuming that sick days are associated with a loss of productivity, this analysis suggests that SIT increases societal welfare. This conclusion also holds if there is no loss of productivity. PMID- 16371216 TI - Total serum IgE: adult reference values in Valencia (1981-2004). Usefulness in the diagnosis of allergic asthma and rhinitis. AB - BACKGROUND: The reference values for total serum IgE are accepted to vary according to the geographical area involved. In 1981 we published the reference values for a donor population in the city of Valencia (Spain) and its metropolitan area. Since then, we have noted an increase in the prevalence of asthma and rhinitis in countries with a Western lifestyle, including Spain. AIM: To update the adult reference values for total serum IgE in the city of Valencia and its metropolitan area, and to determine their diagnostic usefulness in asthma and allergic rhinitis. MATERIAL AND METHODS: We studied two groups of 69 and 100 individuals each. One group comprised apparently healthy blood donors, as assessed by a routine blood bank study protocol and the application of a questionnaire to exclude perceived allergic or parasitic diseases, while the other group consisted of individuals with atopic respiratory disease (bronchial asthma and allergic rhinitis) studied in our service in the same time period (spring 2004). Total serum IgE was determined by the UniCAP method developed by Pharmacia. RESULTS: The geometric mean of total serum IgE among blood donors was 46.65 kU/l (95% CI: 15.5-77.8) versus 204.29 kU/l (95% CI: 93.3-515) among allergic subjects. The difference between the two groups was statistically significant. No significant difference was found between our donor sample in 1981 and the sample in the present study. Establishing an arbitrary cut-off point of 1.65 standard deviations (SD)(equivalent to 95 % of the donor population), we obtained a figure of > 183 kU/l whereby 44% of all allergic individuals presented a value below this cut-off point. To practical effects, these allergic patients behaved as low responders (LRs) in relation to total IgE, whereas the remaining individuals were high responders (HRs) in relation to total IgE. CONCLUSIONS: Our data confirm the current validity of the reference values obtained in 1981, which continue to be significantly different from those of the UniCAP method for Swedish donors. The population of allergic subjects can be divided into two subgroups: subjects with total IgE levels similar to those of the donors, which we refer to as LRs, and which represent 44% of all allergic subjects, and a second subgroup with total IgE values above those of the donors (HRs), who would therefore be the only subjects that would be identified by individual serum IgE study. PMID- 16371217 TI - Rebound phenomenon to systemic corticosteroid in atopic dermatitis. AB - Three patients with atopic dermatitis, one boy and two girls, aged between 6 and 17 years, presented eczematous skin, pruritus, scarifications, lichenification and a family history of atopy. During exacerbations, the patients sought emergency care and were prescribed oral corticosteroids for a period of approximately 15 days. Initially, the patients improved but after cessation of therapy or dose reduction, marked worsening occurred with the development of lesions with extreme pruritus, several confluent lesions, scarification and intense exudates, as well as fever and dehydration. The patients' condition was so severe that two were admitted to the allergy unit. The medication was withdrawn and intravenous hydration was administered, together with hydrating skin creams and antihistamine therapy. In addition, weak topical corticosteroids were applied on the most severely affected areas. All three patients progressively improved. We conclude that the patients with atopic dermatitis described herein presented a rebound phenomenon after the use of corticosteroids. We believe that systemic corticosteroids may exacerbate the acute phase of atopic dermatitis, mediated by IgE, accentuating the Th2 pattern in these patients. PMID- 16371218 TI - Food-protein-induced enterocolitis syndrome caused by fish. AB - INTRODUCTION: Food protein-induced enterocolitis syndrome (FPIES) occurs in infants. Typical symptoms include profuse vomiting and/or diarrhea several hours after ingestion of a given food. The disorder is a non-IgE mediated food hypersensitivity. The most frequently involved foods are milk and soy, but some cases of FPIES induced by solid foods have been described. We report 14 patients with FPIES due to fish protein. MATERIAL AND METHODS: History and physical examination, skin prick test (SPT) with fish allergens and Anisakis simplex, prick-by-prick test with implicated fish and determination of specific IgE antibodies against fish were performed. In eight children atopy patch test (APT) were also performed. In nine patients an open oral food challenge with the implicated fish was carried out. RESULTS: There were six boys and eight girls, aged from 9 to 12 months at diagnosis, with between two and six reactions to the offending fish proteins before the diagnosis was established. Four patients had a previous history of atopy. Presenting symptoms included diarrhea in two patients, profuse vomiting in six patients, and recurrent vomiting and subsequent diarrhea in three patients. In addition to these symptoms, associated septic appearance, apathy and lethargy were present in the remaining three patients. Onset of symptoms occurred a few minutes after fish ingestion in two patients and from 60 minutes to 6 hours in the 12 remaining patients. SPT to fish were negative in all patients. Serum food-specific IgE antibodies were negative in all patients except one. APT was positive in three patients. Open oral challenge (OC) was performed in nine infants and was positive in all. The patients were followed-up for between 1 and 7 years after diagnosis, and follow-up OC tests were performed after fish had been eliminated from the patients' diet for 3-4 years. Four patients became clinically tolerant to the causal food. Three patients currently tolerate only one type of fish (swordfish). CONCLUSIONS: We report 14 patients with FPIES caused by fish protein. The symptoms suggest a form of cell-mediated, non-IgE mediated food hypersensitivity. The gold standard for diagnosis is OC, although caution should be exercised in infants with several reactions or a recent diagnosis. After a period of elimination of the causal food from the diet, tolerance can develop. PMID- 16371219 TI - How pediatricians in Spain manage the first acute wheezing episode in an atopic infant. Results from the TRAP study. AB - BACKGROUND: Although the treatment of asthma has been addressed in several guidelines, the management of the first acute wheezing episode in infants has not often been evaluated. We surveyed practicing pediatricians in Spain about the treatment they would provide in a simulated case. MATERIAL AND METHODS: A random sample of 3000 pediatricians and physicians who normally treated children was surveyed. The questionnaire inquired about how they would treat a first mild-to moderate wheezing attack in a 5-month-old boy with a personal and family history of allergy. Pediatricians were asked about their professional background. RESULTS: A total of 2347 questionnaires were returned with useful data (78.2%). Most (90.4%) of the pediatricians would use a short-acting beta2 agonist (SABA) via a metered-dose inhaler with a spacer and a face mask or nebulizer. However, only 34.5% chose a SABA alone: 31.3% added an oral steroid and 27.6% added an inhaled corticosteroid (ICS). The factors associated with the use of ICS in the acute attack were: (1) lack of specific training in pediatrics (OR 1.45; 1.12 1.85) and (2) primary care health center setting (OR 1.31; 1.01-1.69) or rural setting (OR 1.28; 1.01-1.66). Forty-four percent did not recommend any follow-up treatment while 20.7% prescribed ICS as maintenance therapy. The factors related to this decision were the same as those described above. CONCLUSIONS: The management of a first wheezing episode seems to meet published guidelines among Spanish pediatricians with formal training in pediatrics and in those who work in a hospital setting or in urban areas. PMID- 16371220 TI - Occupational rhinoconjunctivitis and asthma by exposure to Lathyrus sativus flour. AB - We report the case of a 42-year-old non-smoking man, who had worked as a carpenter for 6 years and who reported a history of rhinorrhea, paroxysmal sneezing, nasocular pruritus, lacrimation, wheezing and dyspnea attacks while preparing a mixture to seal the junctures between wooden panels. Allergy study consisted of skin prick testing (SPT) to inhalants, foods and Lathyrus sativus flour (LSF) extract, specific bronchial provocation test with LSF extract, cytological analysis of sputum, specific IgE antibodies against LSF, and histamine releasing test with dilutions (1:5, 1:25, 1:125, 1:625) of LSF. The results demonstrated occupational rhinoconjunctivitis and asthma due to LSF exposure. We provide a review of published reports to date. PMID- 16371221 TI - Delayed hypersensitivity reactions to corticosteroids. AB - BACKGROUND: Corticosteroids (CS) are widely used in the treatment of asthma, allergic disorders and other immunological diseases due to their anti inflammatory and immunosuppressive properties. Physicians seldom suspect them of causing allergic reactions. However, more and more cases of hypersensitivity reactions to CS have been described. Reports of delayed allergic reactions to CS in patients with asthma or allergic rhinitis are scarce. MATERIAL AND METHODS: We report the case of a 44-year-old woman with a history of mild persistent asthma and intermittent allergic rhinitis, treated with inhaled beclomethasone and salbutamol, who developed a delayed mucocutaneous and respiratory reaction after substitution of beclomethasone with budesonide. CONCLUSIONS: The interest of this case lies in the rarity of allergic reactions from inhaled CS in patients with asthma and/or rhinitis. These reactions therefore represent a diagnostic and therapeutic challenge. PMID- 16371222 TI - Vasculitis during immunotherapy treatment in a patient with allergy to Cupressus arizonica. AB - Allergen immunotherapy dates back to 1911 and has been used successfully to treat large numbers of patients throughout the last century. CASE REPORT: a 66-year-old woman presented with symptoms of allergic rhinitis and asthma due to sensitization to Cupressus arizonica. Specific immunotherapy was prescribed as a continuous 2-year treatment with a depot preparation of standarized and characterized allergen extracts of Cupressus arizonica pollen. Forty-eight hours after one maintenance dose of 0.8 cc, the patient presented palpable violaceous purpuric lesions and pruritus on both legs. We performed skin prick and intradermal tests with Cupressus arizonica. Twenty-four hours later, the 1/1 dilution intradermal skin test was positive. Biopsy showed leukocytoclastic vasculitis. CONCLUSIONS: A middle-aged woman experienced cutaneous non necrotizing vasculitis after 2 years of maintenance immunotherapy. The interval between injections and the first appearance of cutaneous lesions suggests a type III hypersensitivity immune reaction. Skin biopsy of the positive intradermal test also supports this hypothesis. PMID- 16371224 TI - Altered cardiac expression of peroxisome proliferator-activated receptor-isoforms in patients with hypertensive heart disease. AB - OBJECTIVE: To investigate whether cardiac expression of the nuclear peroxisome proliferator-activated receptor alpha (PPARalpha) is altered in patients with hypertensive heart disease (HHD). METHODS: We studied endomyocardial septal biopsies from 24 patients with essential hypertension divided into three groups: 6 without left ventricular hypertrophy (LVH) (HT group), 10 with LVH (LVH group), and 8 with LVH and heart failure (HF) (HF group). The expression of two PPARalpha isoforms (the native active and the truncated inhibitory) was analyzed by Western blot and reverse transcription polymerase chain reaction (RT-PCR), and two PPARalpha target genes were evaluated by RT-PCR. Histomorphological features were evaluated in a second myocardial sample from LVH and HF groups. RESULTS: Whereas the expression of native PPARalpha protein was lower (p<0.05) in LVH and HF groups than in the HT group, truncated PPARalpha protein was overexpressed (p<0.001) in the HF group as compared with LVH and HT groups. The mRNA expression of native and truncated PPARalpha was similar in the three groups of hypertensives. In addition, a progressive decrease (p for trend<0.05) in the two PPARalpha target genes mRNA expression was observed among HT, LVH and HF groups. The amount of truncated PPARalpha protein correlates directly with cardiomyocytes apoptosis and inversely with cardiomyocytes density in patients with HHD. In addition, the expression of truncated PPARalpha protein was directly correlated with left ventricular volumes, and inversely with ejection fraction in all hypertensives. CONCLUSIONS: These findings suggest that post-transcriptional regulation of PPARalpha isoforms is altered in patients with HHD, namely in those developing HF. An excess of the truncated inhibitory isoform may be involved in hypertensive left ventricular failure and remodeling. PMID- 16371225 TI - Purine P1 receptor-dependent immunostimulatory effects of antiviral acyclic analogues of adenine and 2,6-diaminopurine. AB - Acyclic nucleoside phosphonates are widely recognised antivirals. The oral prodrugs of prototype compounds, e.g., 9-[2-(phosphonomethoxy)ethyl]adenine (PMEA; adefovir), and 9-(R)-[2-(phosphonomethoxy)propyl]adenine [(R)-PMPA; tenofovir] were approved by FDA for treatment of hepatitis B (Hepsera), and acquired immunodeficiency syndrome (AIDS) (Viread), respectively. A number of acyclic nucleoside phosphonates possess immunostimulatory activity. The present experiments demonstrate that activation of cytokine and chemokine secretion is mediated by adenosine receptors. Included in the study were 9-(R)-[2 (phosphonomethoxy)propyl]adenine [tenofovir], N(6)-cyclopentyl-(R)-9-[2 (phosphonomethoxy)propyl]-2,6-diaminopurine, N(6)-cyclopropyl-(R)-9-[2 (phosphonomethoxy)propyl]-2,6-diaminopurine, and N(6)-isobutyl-9-[2 (phosphonomethoxy)ethyl]-2,6-diaminopurine. All of them activate secretion of tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), "regulated on activation of normal T cell expressed and secreted" (RANTES/CCL5), and macrophage inflammatory protein-1alpha (MIP-1alpha/CCL3) in murine macrophages. With exception of MIP-1alpha, the effects were inhibited by antagonists of adenosine A(1), A(2B), and A(3) receptors (not by adenosine A(2A) receptor antagonist). The adenosine A(1) receptor antagonist inhibited TNF-alpha, IL-10, and RANTES, adenosine A(2B) receptor antagonist inhibited TNF-alpha and RANTES, and adenosine A(3) receptor antagonist inhibited IL-10 and RANTES. The suppression is due to decreased transcription of cytokine mRNA. It may be suggested that acyclic nucleoside phosphonates are nonspecific ligands for purine P(1) receptors. PMID- 16371226 TI - Stimulation of soluble guanylyl cyclase by BAY 41-2272 relaxes anococcygeus muscle: interaction with nitric oxide. AB - The compound BAY 41-2272 stimulates the soluble guanylyl cyclase in a nitric oxide (NO)-independent manner. We have investigated the potency and efficacy of BAY 41-2272 in the rat anococcygeus muscle, as well as the effects of BAY 41-2272 on NO-mediated anococcygeus relaxations. BAY 41-2272 (0.01-10 microM) potently relaxed precontracted anococcygeus muscle strips, with a pEC(50) value of 6.44 +/ 0.03 and maximum response of 100 +/- 2%. The soluble guanylyl cyclase inhibitor 1H-[1,2,4]-oxidiazolo[4,3-a] quinoxalin-1-one (ODQ, 1 microM) and the NO inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 microM) caused significant rightward shifts in the concentration-response curves to BAY 41-2272. The phosphodiesterase type-5 inhibitor tadalafil (0.1 microM) markedly enhanced the relaxations evoked by BAY 41-2272. In addition, BAY 41-2272 increased the duration of nitrergic relaxations by approximately 55%. The relaxations induced by glyceryl trinitrate were also significantly potentiated by BAY 41-2272. In conclusion, BAY 41-2272 interacts with endogenous and exogenous NO causing a potent relaxation of rat anococcygeus muscle. PMID- 16371227 TI - In vitro and in vivo vasodilator activity of racemic tramadol and its enantiomers in Wistar rats. AB - Tramadol ((+/-)-tramadol) is an analgesic agent formulated as a racemic mixture (1:1) of (-)- and (+)-tramadol, which differ in their potency to bind to mu opioid receptors and to inhibit monoamine-reuptake. We investigated the stereoselectivity of in vitro tramadol-induced vasodilatation of aortic rings and its effect on the arterial blood pressure measured in conscious Wistar rats. (+) Tramadol, but not (-)-tramadol, produced a concentration-dependent relaxation of aorta precontracted with phenylephrine. The concentration-response curve was significantly altered by the removal of endothelium. Vascular relaxation was also inhibited by pre-incubation of endothelium-intact aorta with naloxone, suggesting the involvement of opioid receptors. The vasodilatation produced by tramadol was stereoselective, and the (+)-tramadol-induced vasodilatation was mediated by mu opioid receptors and partially dependent on endothelium integrity. The hypotensive response induced by (+)-tramadol was also observed after bolus injection of 5.0 and 10.0 mg/kg. The results indicate that only high doses of tramadol cause cardiac depression and hypotension, indicating that it can be used safely. PMID- 16371228 TI - Oral misoprostol for prevention of postpartum hemorrhage by paramedical workers in India. AB - OBJECTIVE: To study whether paramedical workers from rural primary health centers in India are able to administer oral misoprostol and actively manage the third stage of labor to prevent postpartum hemorrhage (PPH). METHOD: Cluster randomization was used to enroll 1200 women at 30 peripheral health centers from 5 states in India, 600 forming the study's intervention group (active management of the third stage of labor with 600 mug of oral misoprostol) and 600 forming the comparison group (in which the current government guidelines for the prevention of PPH were followed). The primary outcome was blood loss after delivery, which was measured using a calibrated blood collection drape. RESULTS: Age, literacy level, occupation, and gravidity were similar in the 2 groups. More than 70% of women in both groups had moderate anemia (hemoglobin level <10 g/dL). Paramedical workers followed instructions in almost all deliveries in the intervention group (99%). There was a significant reduction in duration of the third stage of labor (7.9 +/- 4.2 min vs. 10.9 +/- 4.3 min; p < .001) and median blood loss after delivery (100 mL vs. 200 mL; p < .001) in the intervention group. Overall, a low incidence of PPH was observed (<1%) in both groups. A greater number of women had moderate to severe shivering (12.7% vs. 0.5%) and a temperature higher than 38 degrees C (9.7% vs. 4.3%) in the intervention group, which was statistically significant. CONCLUSION: Simple interventions can be easily implemented in rural health care settings to reduce the blood loss during labor. This finding has significant implications for developing countries, in which the prevalence of anemia is high. PMID- 16371229 TI - Protection against dextran sodium sulfate-induced colitis by dehydroepiandrosterone and 7alpha-hydroxy-dehydroepiandrosterone in the rat. AB - In this study the anti-oxidant effect of DHEA and 7alpha-hydroxy-DHEA against oxidative stress induced by colitis was investigated in vivo in rats. The two steroids were intraperitoneally injected once daily (50 mg/kg body weight) for 7 days before the induction of colitis that was effected by a daily treatment of 5% (w/v) dextran sodium sulfate (DSS) in drinking water for 7 days. This was quantified by the evidence of weight loss, rectal bleeding, increased wall thickness, and colon length. The inflammatory response was assessed by neutrophil infiltration after a histological examination and myeloperoxidase (MPO) activity measurement. Two markers of oxidative damage were measured in colon homogenates after the onset of DSS treatment: protein carbonyls and thiobarbituric acid reacting substances. The colonic metabolism of corticosterone by 11beta hydroxysteroid dehydrogenases types 1 and 2 (11beta-HSD) was investigated in control and treated animals. Results indicated that colitis caused a decrease in body weight and colon length. Severe lesions were observed in the colon with a reduced number of goblet cells which contained less mucins. The lesions were associated with increased MPO activity and oxidative damage. Colonic inflammation down and up regulated the 11beta-HSD2 and 11beta-HSD1, respectively. Treatments by DHEA and 7alpha-hydroxy-DHEA attenuated the inflammatory response when MPO activity decreased; but this did not increase the colonic oxidation of corticosterone into 11-dehydrocorticosterone. Both DHEA and 7alpha-hydroxy-DHEA exerted a significant anti-oxidant effect against oxidative stress induced by colitis through reducing the oxidative damage to proteins and lipids. This resulted in a moderate increase in the amount of colonic mucus. Both DHEA and 7alpha-hydroxy-DHEA may prove useful in the prevention or treatment of colitis. PMID- 16371230 TI - Characteristics of nitrate reduction by zero-valent iron powder in the recirculated and CO(2)-bubbled system. AB - In this study, the Fe(0)/CO(2) process was investigated for removing nitrate from aqueous solution under different operating conditions such as CO(2) bubbling rate (0-400 mL/min), Fe(0) dosage (1-6g/L), initial nitrate concentration (6-23 mgN/L), batch mode, and fresh Fe(0) supplementing (0-1g/L). Results show that the bubbling of CO(2) flow rate at 200 mL/min was sufficient for supplying H(+) into solution to create an acidic environment favorable to nitrate reduction reaction. It was found that sigmoidal model equation describes the S-curve behaviors of nitrate reduction, ferrous accumulation and ammonium formation satisfactorily, and the parameter t(1/2) of the proposed model equation serves as a powerful tool for the comparison of nitrate reduction rate. Sustainability test demonstrates that Fe(0) powder began to deteriorate after three batches operation. Concerning the operating modes, the batch mode with the treated solution emptied and freshly refilled outperforms the one, which was operated by retaining the treated solution and spiking concentrated nitrate into it for the next batch treatment. To guarantee satisfactory nitrate removal using the former mode, supplement of appropriate amount of Fe(0) needs to be optimized. PMID- 16371231 TI - Three new arsenic hyperaccumulating ferns. AB - Phytoremediation, an emerging, plant-based technology for the removal of toxic contaminants from soil and water, has been receiving increased attention. The prerequisite for successful phytoremediation is the existence of hyperaccumulator plants. Designed to search for new arsenic (As) hyperaccumulators, an experiment was conducted under greenhouse conditions in a completely randomized design with four replications. This experiment identified Pteris biaurita L., P. quadriaurita Retz and P. ryukyuensis Tagawa as new hyperaccumulators of As and re-confirmed Pteris cretica as a hyperaccumulator. The average As concentration ranged from 1770 to 3650 mg kg(-1) DW in the fronds and 182 to 507 mg kg(-1) DW in the roots of P. cretica, P. biaurita, P. quadriaurita and P. ryukyuensis after having been grown in 100 mg As kg(-1) soil. There was a greater percentage of As(III) as compared to As (V) in the fronds of these plants. Based on our study, P. ryukyuensis is the most promising candidate to phytoremediate As contaminated soils compared to the other three species. The nutrient requirements or distributions within the Pteris species were altered distinctly when the plants were exposed to As. PMID- 16371232 TI - Phenotype correction of hemophilia A mice with adeno-associated virus vectors carrying the B domain-deleted canine factor VIII gene. AB - Adeno-associated virus (AAV) vectors carrying the B domain-deleted canine FVIII (BDD cFVIII) gene utilizing the beta-actin minimum promoter (167b) pseudotyped with serotype 1 (AAV1-beta-actin-cFVIII) and serotype 8 (AAV8-beta-actin-cFVIII) were developed to express cFVIII in hemophilia A mice. FVIII clotting activities measured by the APTT method increased in hemophilia A mice with intramuscular injection of AAV1-beta-actin-cFVIII in a dose-dependent manner. Therapeutic FVIII levels (2.9+/-1.0%) in hemophilia A mice with the AAV1-beta-actin-cFVIII dose of 1 x 10(12) gc/body were achieved, suggesting partial correction of the phenotype with AAV1-beta-actin-cFVIII vectors. FVIII clotting activity levels in hemophilia A mice with intravenous injection of AAV8-beta-actin-cFVIII also were increased dose-dependently, achieving therapeutic FVIII levels (5-90%) in hemophilia A mice with the AAV8-beta-actin-cFVIII doses of 1-3 x 10(11) gc/body and supernormal FVIII levels (180-670%) in hemophilia A mice with the AAV8-beta-actin-cFVIII dose of 1 x 10(12) gc/body. Transduction of the liver with AAV8-beta-actin-cFVIII is superior to transduction of skeletal muscles with AAV1cFVIII regarding the FVIII production and antibody formation. These data suggested that both AAV1 and AAV8 vectors carrying the FVIII gene utilizing a minimum promoter have a potential for hemophilia A gene therapy. PMID- 16371233 TI - Effects of vitamin D analogs on the expression of plasminogen activator inhibitor 1 in human vascular cells. AB - INTRODUCTION: Vitamin D analogs such as paricalcitol and calcitriol have been shown to provide survival benefit for Stage 5 chronic kidney disease (CKD) patients, possibly due to their positive impact on the cardiovascular system. Plasminogen activator inhibitor-1 (PAI-1) is one of the risk markers for coronary artery disease. MATERIALS AND METHODS: Human coronary artery smooth muscle cells (SMC) and endothelial cells (CAEC) were treated with vitamin D analogs to assess the effects of the drugs on the expression of PAI-1 mRNA and protein. RESULTS: In SMC, both paricalcitol and calcitriol down-regulated the expression of PAI-1 mRNA and protein in a dose-dependent manner. The EC(50) values of paricalcitol and calcitriol on suppressing PAI-1 mRNA were 3.0 and 2.8 nM, respectively. Interestingly, these two drugs had no significant effect on the expression of PAI 1 protein or mRNA in CAEC. Further analysis showed that CAEC did not express functional vitamin D receptor (VDR) and paricalcitol failed to induce the expression of 25-hydroxyvitamin D-24-hydroxylase (CYP24A1) mRNA, a gene known to be regulated by VDR. As a comparison, SMC expressed VDR and paricalcitol induced CYP24A1 mRNA in SMC (>150-fold at 10 nM) dose-dependently. The effect of paricalcitol on suppressing PAI-1 in SMC was blocked by cycloheximide, suggesting that protein synthesis was involved. CONCLUSION: These results demonstrate that vitamin D analogs suppress PAI-1 in SMC, but not in CAEC. Suppression of PAI-1 in SMC may be one of the factors contributing to the survival benefits of vitamin D analog therapy in CKD patients. PMID- 16371234 TI - Involvement of COX-1 and up-regulated prostaglandin E synthases in phosphatidylserine liposome-induced prostaglandin E2 production by microglia. AB - After engulfment of apoptotic cells through phosphatidylserine (PS)-mediated recognition, microglia secrete prostaglandin E2 (PGE2), a potent anti inflammatory molecule in the central nervous system. Despite the clinical significance, the mechanism underlying PGE2 production by phagocytosis of apoptotic cells is poorly understood. In the present study, we used PS liposomes to elucidate the phagocytic pathway for PGE2 production in microglia, because PS liposomes mimic the effects of apoptotic cells on microglia/macrophages. The level of PGE2 in the culture medium of primary cultured rat microglia was significantly increased by PS liposomes treatment but not by phosphatidylcholine liposomes treatment. The specific ligand for class B scavenger receptor (SR-B), high density lipoprotein, significantly suppressed PS liposome-induced PGE2 production. PS liposomes were immediately phagocytosed by microglia and sorted to endosomes/lysosomes. Cyclooxygenase (COX)-2 and membrane-bound prostaglandin E synthase-1 (mPGES-1) were induced by treatment with lipopolysaccharide (LPS) but not with PS liposomes. On the other hand, mPGES-2 and cytosolic PGES (cPGES) that are functionally coupled with COX-1 were upregulated after treatment with PS liposomes or LPS. Furthermore, PS liposome-induced PGE2 production was significantly suppressed by indomethacin, a preferential COX-1 inhibitor, but not by NS-398, a selective COX-2 inhibitor. PS liposomes induced activation of p44/p42 extracellular signal-regulated kinase (ERK) but not p38 mitogen-activated protein kinase in SR-BI independent manner. These observations strongly suggest that the up-regulation of terminal PGESs that are preferentially coupled with COX 1, especially mPGES-2, plays the pivotal role in PS liposome-induced PGE2 production by microglia. Although SR-BI plays an essential role in PS liposome induced PGE2 production, other PS-recognizing receptors, possibly PS-specific receptor, could also promote PGE2 production by transducing intracellular signals including p44/p42 ERK after PS liposomes treatment. PMID- 16371235 TI - Specificity of recombinant antibodies generated from multiple sclerosis cerebrospinal fluid probed with a random peptide library. AB - We generated recombinant antibodies (rAbs) from over-represented IgG sequences expressed by single plasma cells from multiple sclerosis (MS) cerebrospinal fluid (CSF). Panning of a phage-displayed random peptide library with the rAbs revealed several specific peptide sequences. Inhibition assays confirmed specific binding of the peptides to the antigen-binding site of the antibody. The native IgG of MS CSF from which the recombinant antibody was cloned also recognized these peptides. Our data demonstrate that MS rAb reflects the specificity of IgG in the CSF. Thus, the epitopes/mimotopes identified by MS rAb may provide clues to disease-relevant antigens. PMID- 16371236 TI - The 2002 European registry of cardiac catheter interventions. AB - OBJECTIVE: The tradition of yearly reports on cardiac catheter interventions in Europe has been initiated in 1992. This 11th report presents aggregated data on cardiac catheter procedures in 30 European countries in the year 2002. DESIGN AND SETTING: A detailed questionnaire addressing summary data of all cardiac interventions was mailed to presidents or delegates of the national societies of cardiology in Europe. The questionnaire was distributed to all institutions with cardiac catheterisation programs. All questionnaires were compiled in a national summary data sheet, then entered into a central database and subsequently analysed. MAIN OUTCOME MEASURES: Coronary angiography, PTCA, and stenting in absolute numbers and per million inhabitants in the participating countries and the whole of Europe. RESULTS: Overall, 1,901,932 coronary angiograms were reported. The population-adjusted rate of coronary angiograms amounted to an absolute mean of 3358 per 10(6) inhabitants, an increase of 7% compared with 2001. A total of 686,869 PTCA procedures were reported. The mean European number of PTCAs per 10(6) inhabitants increased by 10% from 1103 in 2001 to 1213 in 2002. Procedures with stenting increased by 17% from 508,999 to 593,906. The stenting rate was 86% compared with 82% in 2001. CONCLUSIONS: In pace with epidemiological demand and the need to catch-up from underuse in certain countries, a continuous and considerable growth of coronary interventions can be observed. It will take years to find out whether the announced change of paradigm in the treatment of multivessel disease in the wake of drug-eluting stents will come true. PMID- 16371237 TI - Early heart rate recovery after exercise predicts mortality in patients with chronic heart failure. AB - BACKGROUND: Patients with chronic heart failure (CHF) have multiple abnormalities of autonomic regulation that have been associated to their high mortality rate. Heart rate recovery immediately after exercise is an index of parasympathetic activity, but its prognostic role in CHF patients has not been determined yet. METHODS: Ninety-two stable CHF patients (83M/9F, mean age: 51+/-12 years) performed an incremental symptom-limited cardiopulmonary exercise testing. Measurements included peak O2 uptake (VO2p), ventilatory response to exercise (VE/VCO2 slope), the first-degree slope of VO2 for the 1st minute of recovery (VO2/t-slope), heart rate recovery [(HRR1, bpm): HR difference from peak to 1 min after exercise] and chronotropic response to exercise [%chronotropic reserve (CR, %)=(peak HR-resting HR/220-age-resting HR)x100]. Left ventricular ejection fraction (LVEF, %) was also measured by radionuclide ventriculography. RESULTS: Fatal events occurred in 24 patients (26%) during 21+/-6 months of follow-up. HRR1 was lower in non-survivors (11.4+/-6.4 vs. 20.4+/-8.1; p<0.001). All cause mortality rate was 65% in patients with HRR112 bpm (log-rank: 32.6; p<0.001). By multivariate survival analysis, HRR1 resulted as an independent predictor of mortality (chi2=19.2; odds ratio: 0.87; p<0.001) after adjustment for LVEF, VO2p, VE/VCO2 slope, CR and VO2/t slope. In a subgroup of patients with intermediate exercise capacity (VO2p: 10 18, ml/kg/min), HRR1 was a strong predictor of mortality (chi2: 14.3; odds ratio: 0.8; p<0.001). CONCLUSIONS: Early heart rate recovery is an independent prognostic risk indicator in CHF patients and could be used in CHF risk stratification. PMID- 16371238 TI - Low dose amiodarone-induced lung injury. AB - Amiodarone-induced lung injury is a potentially lethal side-effect of amiodarone. In an attempt to assess its risk, we studied the incidence and clinical profile of Chinese patients prescribed with low dose amiodarone. PMID- 16371239 TI - Rapid identification of bacteria by real-time amplification and sequencing of the 16S rRNA gene. AB - To allow rapid identification of bacteria in pure cultures and blood culture bottles, an assay was developed which is based on real-time amplification and sequencing of bacterial 16 S rRNA genes. In principle, this assay allows identification of bacteria from pure cultures within 6.5 h, and from blood cultures within approximately 7 h. PMID- 16371240 TI - An efficient and generic extension to ITK to process arbitrary shaped regions of interest. AB - The paper describes a software method to extend ITK (Insight ToolKit, supported by the National Library of Medicine), leading to ITK++. This method, which is based on the extension of the iterator design pattern, allows the processing of regions of interest with arbitrary shapes, without modifying the existing ITK code. We experimentally evaluate this work by considering the practical case of the liver vessel segmentation from CT-scan images, where it is pertinent to constrain processings to the liver area. Experimental results clearly prove the interest of this work: for instance, the anisotropic filtering of this area is performed in only 16 s with our proposed solution, while it takes 52 s using the native ITK framework. A major advantage of this method is that only add-ons are performed: this facilitates the further evaluation of ITK++ while preserving the native ITK framework. PMID- 16371241 TI - The RecQ gene family in plants. AB - RecQ helicases are conserved throughout all kingdoms of life regarding their overall structure and function. They are 3'-5' DNA helicases resolving different recombinogenic DNA structures. The RecQ helicases are key factors in a number of DNA repair and recombination pathways involved in the maintenance of genome integrity. In eukaryotes the number of RecQ genes and the structure of RecQ proteins vary strongly between organisms. Therefore, they have been named RecQ like genes. Knockouts of several RecQ-like genes cause severe diseases in animals or harmful cellular phenotypes in yeast. Until now the largest number of RecQ like genes per organism has been found in plants. Arabidopsis and rice possess seven different RecQ-like genes each. In the almost completely sequenced genome of the moss Physcomitrella patens at least five RecQ-like genes are present. One of the major present and future research aims is to define putative plant specific functions and to assign their roles in DNA repair and recombination pathways in relation to RecQ genes from other eukaryotes. Regarding their intron positions, the structures of six RecQ-like genes of dicots and monocots are virtually identical indicating a conservation over a time scale of 150 million years. In contrast to other eukaryotes one gene (RecQsim) exists exclusively in plants. It possesses an interrupted helicase domain but nevertheless seems to have maintained the RecQ function. Owing to a recent gene duplication besides the AtRecQl4A gene an additional RecQ-like gene (AtRecQl4B) exists in the Brassicaceae only. Genetic studies indicate that a AtRecQl4A knockout results in sensitivity to mutagens as well as an hyper-recombination phenotype. Since AtRecQl4B was still present, both genes must have non-redundant roles. Analysis of plant RecQ-like genes will not only increase the knowledge on DNA repair and recombination, but also on the evolution and radiation of protein families. PMID- 16371242 TI - Conceptual, methodological, and analytical issues in the study of relapse. AB - This article examines conceptual, methodological, and analytic issues in research on relapse to alcohol and other drug use. The review notes the continued move in relapse research from a primary reliance on retrospective assessment of factors surrounding the onset of relapse episodes to an increased focus on the use of new technologies to obtain "near real time" data on proximal factors in relapses. Recent advances in neurobiology have yielded important gains in our understanding of vulnerability to relapse in alcohol and other drug abusers. New statistical techniques are also available to analyze data on relapse. From a theoretical standpoint, there has been an increasing appreciation for the complicated and dynamic interplay of distal and proximal factors in the relapse process. At this point, the strongest and most detailed data on factors in the onset and course of relapse have been generated by studies of smoking relapses that have made use of Ecological Momentary Assessment (EMA) technology. However, there is still limited "near real" time data on proximal factors in alcohol and other drug relapses, and no such data on factors that influence the course of these relapses, once they have begun. Nevertheless, important methodological advances have been and continue to be made in the study of relapse, and our knowledge about the nature and process of relapse has increased considerably over the past 10 years. PMID- 16371243 TI - Recycling of an industrial sludge and marine clay as light-weight aggregates. AB - The geographical limitations of Singapore, its restricted natural resources and voluminous municipal and industrial waste streams, make environmental management a major challenge for the island state. In an attempt to find ways to reduce importation of raw materials and the waste sent to landfill, light weight aggregates were produced from marine clay and a CaF(2)-rich semiconductor industry sludge. Aggregates were produced in a bench-scale rotary kiln with three clay/sludge loadings (90/10, 70/30 and 50/50%). All three mixtures showed good bloating behavior during firing and the ceramic pellets (1-1.5cm diameter) had densities well below that required for light-weight aggregates. In the initial tests, the pore sizes of the aggregates were in general too large resulting in high water absorption. Comparisons between the composition of the two waste products and the aggregates showed a significant loss of fluorine (40-60%) during processing; a problem which may require flue gas treatment. Leach testing showed that the formed aggregates would not pose a human or environmental hazard in terms of fluorine mobilization. PMID- 16371244 TI - Effects of natural prenylated flavones in the phenotypical ER (+) MCF-7 and ER ( ) MDA-MB-231 human breast cancer cells. AB - The effect of seven natural prenylated flavones in DNA synthesis of two human breast cancer cell lines, the estrogen-dependent ER (+) MCF-7 and the estrogen independent ER (-) MDA-MB-231 cells, was evaluated. Flavones with an isopentenyl group at C-8 and a ring linking C-3 and C-2' presented a biphasic effect in DNA synthesis of ER (+) MCF-7 and displayed a stimulation at low concentrations (0.02 0.78 microM) whilst at higher concentrations (> 3.12 microM) inhibition was observed. No stimulation was observed in ER (-) MDA-MB-231. In contrast, all the flavones exhibited an antiproliferative effect in both ER (-) and ER (+). Curiously, the inhibition of DNA synthesis was accompanied by a high capacity of these cells to reduce MTT, which was concurrent with the appearance of an intense intracytoplasmic vacuolization. The accumulation of the formazan product in these vacuoles could justify the enhancements of MTT reduction. The characterization of these vacuoles with the autophagic marker monodansylcadaverine (MDC) is consistent with autophagic vacuoles, which led to the suggestion that these flavones could induce autophagy in both ER (+) and ER (-) breast cancer cell lines. PMID- 16371245 TI - Factors of morbidity in hemispherectomies: surgical technique x pathology. AB - OBJECTIVE: The objective of this paper is to evaluate factors of surgical morbidity from different techniques of hemispherectomy with emphasis on causative pathology. PATIENTS AND METHODS: Thirty patients underwent hemispherectomy in our institution from 1987 to 2003, two presented with Sturge-Weber Syndrome (SWS), sixteen with Rasmussen's Syndrome (RS), eight with established hemispheric lesions (EHL), and four with cortical development malformations (CDM). Six surgeons operated on three patients using anatomical hemispherectomies (AH), 11 patients using functional hemispherectomy (FH), and 16 patients employing hemispherotomy (HT). Surgical technique and causative pathology were studied independently as factors of morbidity in hemispherectomy. RESULTS: Overall mean surgical time was 11:50+/-3:20 h and increased proportionately in pathologies with larger hemispheres. Blood transfusion was particularly influenced by the approach adopted by our team of anesthesiologists, independently of technique or pathology. Pathology was the most important factor related to hydrocephalus as two out of four patients with CDM needed ventriculoperitoneal shunt whilst none with EHL or SWS. Four patients undergoing HT and one FH presented residual bridges connecting the hemispheres, three were reoperated and are seizure free. Two patients with CDM did not improve their seizures worthwhile with surgery and other two (one with RS and other with CDM) were waiting a second procedure due to incomplete inter-hemispheric disconnection. Five patients presented infection and one died after developing meningoencephalitis. CONCLUSION: Hemispherectomies are procedures where pathology and surgical technique interact narrowly. Therefore, in order to study surgical morbidity or outcome, both pathology and technique have to be analyzed independently. PMID- 16371246 TI - Mycobacterium xenopi multiplies within human macrophages and enhances HIV replication in vitro. AB - Mycobacterium xenopi can cause opportunistic infections, particularly in persons infected with human immunodeficiency virus type 1 (HIV-1). The primary focus of this effort was to determine if M. xenopi isolates could survive and grow in human peripheral blood macrophage (MPhi), and if these isolates could promote the replication of HIV-1 in vitro. M. xenopi bacilli survived and replicated 10-fold within 48 h in human MPhi while avirulent Mycobacterium smegmatis, did not grow within the MPhi. M. xenopi bacilli when cultured with peripheral blood mononuclear cells enhanced HIV-1 replication 30- and 50-fold with the macrophage tropic HIV-1(Ba-L) and 50- and 75-fold with T-cell-tropic strain HIV-1(LAI) by 6 days post-infection when compared to M. smegmatis. The enhanced HIV replication was associated with increased production of TNF-alpha. Partial inhibition of HIV 1 induction was observed using a neutralizing anti-TNF-alpha monoclonal antibody, pentoxifylline, and matrix metalloproteinase (MMP) inhibitor I. Similar mechanisms of pathogenesis among mycobacterial species may help elucidate better treatment approaches in HIV co-infected persons. PMID- 16371247 TI - TRAF6 and Src kinase activity regulates Cot activation by IL-1. AB - Cot is one of the MAP kinase kinase kinases that regulates the ERK1/ERK2 pathway under physiological conditions. Cot is activated by LPS, by inducing its dissociation from the inactive p105 NFkappaB-Cot complex in macrophages. Here, we show that IL-1 promotes a 10-fold increase in endogenous Cot activity and that Cot is the only MAP kinase kinase kinase that activates ERK1/ERK2 in response to this cytokine. Moreover, in cells where the expression of Cot is blocked, IL-1 fails to induce an increase in IL-8 and MIP-1betamRNA levels. The activation of Cot-MKK1-ERK1/ERK2 signalling pathway by IL-1 is dependent on the activity of the transducer protein TRAF6. Most important, IL-1-induced ERK1/ERK2 activation is inhibited by PP1, a known inhibitor of Src tyrosine kinases, but this tyrosine kinase activity is not required for IL-1 to activate other MAP kinases such as p38 and JNK. This Src kinases inhibitor does not block the dissociation and subsequently degradation of Cot in response to IL-1, indicating that other events besides Cot dissociation are required to activate Cot. All these data highlight the specific requirements for activation of the Cot-MKK1-ERK1/ERK2 pathway and provide evidence that Cot controls the functions of IL-1 that are mediated by ERK1/ERK2. PMID- 16371248 TI - Speech production in schizophrenia: preliminary data regarding voice onset time. PMID- 16371249 TI - Role of neuropeptides in Behcet's disease in relation to clinical activity. PMID- 16371250 TI - Gray matter density reduction in the insula in fire survivors with posttraumatic stress disorder: a voxel-based morphometric study. AB - Voxel-based morphometry (VBM) is an objective whole-brain technique for characterizing regional cerebral volume and tissue concentration differences in structural magnetic resonance images. In the current study, we used VBM to examine possible cerebral gray matter abnormalities in patients with posttraumatic stress disorder (PTSD) due to fire. The subjects included 12 victims of a fire disaster with PTSD and 12 matched victims of the same fire without PTSD. Magnetic resonance images were obtained on a 1.5-Tesla General Electric scanner at Central South University, and an entire brain volume of 248 contiguous slices was obtained for each subject. Then, gray matter density in patients with PTSD and control groups was compared by using a VBM approach in SPM2. Group analysis was thresholded at P<0.001, uncorrected, at the voxel level. The following three regions of reduced gray matter volume were found in patients with PTSD compared with controls: left hippocampus, left anterior cingulate cortex (ACC), and bilateral insular cortex. It was concluded that there are structural abnormalities of the hippocampus, the ACC and the insular cortex in patients with PTSD due to fire. PMID- 16371251 TI - Cancer screening: the importance of outcome measures. AB - Cancer screening is attracting greater attention as a growing number of studies and practice guidelines exhort physicians to screen for an increasing number of malignancies. Central to the evaluation of these studies is both the quality of the evidence and the impact of screening upon clinically relevant outcomes. Although much has been written about the optimal study design for screening studies, surprisingly little has been written about evaluating specific outcome measures employed in such studies. This paper reviews the strengths and limitations of common outcomes employed in cancer screening studies. The relationship between study design and outcome measure is explored. Three key biases (lead-time, length-time, and volunteer) associated with screening studies and methods to minimize them are also reviewed. Potential harms associated with screening are outlined. Although disease-specific mortality may be the most robust and relevant traditional outcome measure, increasing attention is being paid to composite health outcome measures for older populations, including quality of life and cost-based measures. PMID- 16371252 TI - Phylogeny of Taenia: Species definitions and origins of human parasites. AB - Phylogeny is fundamental as it constrains explanations about history and forms our foundation for recognizing and diagnosing species. In the absence of such a framework taxonomists historically relied on intuitive processes, personal judgment and authority, often embracing a typological view of species that disregarded otherwise unequivocal historical and biological criteria. Species of Taenia are among the most characteristic tapeworms infecting carnivores and humans as definitive hosts and indeed Taeniidae is unique among the Eucestoda in requiring 2 obligate mammalian hosts for transmission; a high percentage (>80%) of life cycles have been completely elucidated among the approximately 45 species and nominal subspecies of Taenia. Until recently there had been no comprehensive attempts at reconstruction of a phylogeny among these important parasites. Such analyses have allowed us to explore the origins and evolution of those independent species of Taenia that occur in humans (T. saginata, T. asiatica, and T. solium) and to understand the ecological and historical processes serving as determinants of biogeography and host-association. These studies supported the status of T. asiatica as a valid species and diagnosed a relationship as the sister-species of T. saginata. These conclusions contrasted with a diversity of opinions that would subsume T. asiatica as a subspecies. Recognition of a species constitutes a specific and testable hypothesis, is not an arbitrary decision and is most appropriately assessed in the context of phylogenetic or historical data. Considering macrospecies, a process has been outlined by Brooks and McLennan [Brooks DR, McLennan DA. The nature of diversity: an evolutionary voyage of discovery. University of Chicago Press: Chicago; 2002] as follows: (1) Discovery: a systematist describes the species; (2) Phylogenetic reconstruction; (3) Evaluation I: do sister-species show geographical overlap-are they sympatric or allopatric (use phylogeny+geographical distributions)? (4) Evaluation II: are sister-species reproductively isolated based on information from natural history, ecology and reproductive biology? Species may be viewed in the context of microevolutionary and macroevolutionary processes. For instance, microspecies are defined in ecological time and involve populations and contemporary process that are potentially reversible (reticulate). In contrast, macrospecies as exemplified by T. saginata and T. asiatica are divergent lineages resulting from processes in evolutionary time where an ancestor has undergone a permanent split that is non reversible (non-reticulate). Applying these criteria in evaluation of T. saginata and T. asiatica, it becomes clear that in evolutionary time these represent historical lineages with independent spatial and temporal trajectories, having separated from a common ancestor near 0.78 to 1.71 MYBP in Africa, or Eurasia. In ecological time, sympatry, reproductive isolation, and differences in life history evident for T. saginata and T. asiatica as observed in China, and perhaps other regions of Southeast Asia, further serve to validate these taeniids. PMID- 16371253 TI - Spatial modelling and ecology of Echinococcus multilocularis transmission in China. AB - Recent research in central China has suggested that the most likely transmission mechanism for Echinococcus multilocularis to humans is via domestic dogs which are allowed to roam freely and hunt (infected) small mammals within areas close to villages or in areas of tented pasture. This assertion has led to the hypothesis that there is a landscape control on transmission risk since the proximity of suitable habitat for susceptible small mammals appears to be the key. We have tested this hypothesis in a number of endemic areas in China, notably south Gansu Province and the Tibetan region of western Sichuan Province. The fundamental landscape control is its effect at a regional scale on small mammal species assemblages (susceptible species are not ubiquitous) and, at a local scale, the spatial distributions of small mammal populations. To date the research has examined relationships between landscape composition and patterns of human infection, landscape and small mammal distributions and recently the relationships between landscape and dog infection rates. The key tool to characterize landscape is satellite remote sensing and these data are used as inputs to drive spatial models of transmission risk. This paper reviews the progress that has been made so far in spatial modeling of the ecology of E. multilocularis with particular reference to China, outlines current research issues, and describes a framework for building a spatial-temporal model of transmission ecology. PMID- 16371254 TI - Electrophysiological correlates of cortico-subcortical interaction: a cross frequency spectral EEG analysis. AB - OBJECTIVE: Several studies have provided evidence for the notion that the coupling between slow and fast frequency in the EEG spectrum indicates cortico subcortical cross-talk (CSC-ct). In addition, findings for increased limbic activation due to reduced cortical inhibition have recently been acquired. To get further insights into these mechanisms, the current study investigated whether CSC-ct would decrease as a function of increased slow (SW) or fast wave (FW) activity. METHODS: Resting state EEG recordings were obtained from 46 healthy, right-handed participants. CSC-ct was quantified by computing cross-frequency correlations between the power in the slow and fast frequency range. CSC-ct was compared between groups with relatively low and high SW activity and groups with relatively low and high FW activity. RESULTS: Relatively reduced SW, but not FW activity was associated with a significant coupling between slow and fast frequency EEG. Furthermore, relatively enhanced resting state SW activity was paralleled by slow and fast frequency EEG decoupling. CONCLUSIONS: These findings are in line with the notion that increased subcortical drive can go accompanied by reduced CSC-ct. SIGNIFICANCE: Cross-frequency EEG analyses might provide a unique approach to obtain novel insights into cortico-subcortical interactions in relation to affective and cognitive behavior. PMID- 16371255 TI - Two centuries of growth among Czech children and youth. AB - The trend of increasing height can be interpreted as a reflection of the unfolding progress of civilization. Height changes among children and adolescents are good markers of this trend. We analyze the secular trend in the heights of children and adolescents in the Czech Republic on the basis of data from anthropological surveys. The earliest height data pertain to Czech youths who attended the Military Schools in Austria in 1800-1809. Data also exist for 1895 and continue in 1951 and at 10-year intervals thereafter. Growth curves were obtained for separate age groups by fitting mean values via third-order polynomial smoothing splines. Between 1951 and 2001, the mean heights of boys and girls aged 2.5 years increased by 2.7 and 3 cm, respectively. Since 1895, the mean height of 13-year-old boys has increased by 19.4 cm, and the mean height of girls has increased by 18.3 cm. PMID- 16371256 TI - Inequality in Japan (1892-1941): physical stature, income, and health. AB - This paper investigates the relationship between physical stature, per capita income, health, and regional inequality in Japan at the prefecture-level for the period 1892-1941. The analysis shows that inequality in income and access to health services explains differences in average height of the population across the 47 Japanese prefectures during this period and that variation in income contributed to changes in height during the 1930s. Annual regional time series of height indicate that Japan experienced a regional convergence in biological welfare before 1914, and that a divergence occurred during the interwar period; personal inequality followed a similar pattern. PMID- 16371257 TI - Editorial. PMID- 16371258 TI - Substance abuse and dependency as a risk factor for delinquency and violent behaviour in schizophrenic patients--how strong is the evidence? AB - There is increasing evidence that schizophrenics are at a comparatively high risk for delinquency and violent behaviour. There is a need to define risk factors for aggression and violence in schizophrenics. Apart from an apparently higher rate of violent and assaultive behaviour in male schizophrenics, on the psychopathological level, symptoms such as vivid hallucinations or delusions have been proposed as possible risk factors. It is possible that substance abuse in schizophrenia may be considered as a risk factor for delinquency and violent behaviour. There is evidence from both clinical and epidemiological studies that this is the case. These data are reviewed. PMID- 16371259 TI - Forensic medicine: education and practice in Japan. PMID- 16371260 TI - The stress of child sexual abuse examinations. AB - The objective of this paper was to determine if the medical evaluation of sexual abuse victims is stressful by heart rate variability monitoring. The design of the study was a case series of children referred for sexual abuse examination to determine child stress response monitored by heart rate variability during baseline, disclosure, and anogenital examination with photographs. The setting was a referral center for the investigation of child sexual abuse in two sites (Reno and Las Vegas, Nevada, USA). Patients included a consecutive sample of 30 children; 15 in Reno ranging in age from 3-10 years with a mean age of 7.1 (1 male and 14 females); 15 in Las Vegas ranging in age from 6-10 years with a mean age of 7.8 (3 males and 12 females). Measurements and results were: each subject's heart rate was obtained during the first 3 minutes of the baseline, disclosure, and the anogenital examination with photographs, using a telemetric Vantage Performance Heart Watch consisting of a 5.5 x 1.25 inch transmitter attached to a chest strap and a microcomputer wristwatch attached to the subject's non-dominant wrist. Heart rates were taken every 5 or 15 seconds throughout the entire examination. A follow-up home visit was made 6 weeks after the examination to determine the replicability of clinic baseline measures and child behavioral sequelae. Physiologic measurements did not show statistically significant stress responses when children were prepared according to the clinic protocol in Reno and Las Vegas. There was a trend toward more responsivity among some of the older subjects during the anogenital examination with photographs, and when clinic procedures inadvertently overlapped several procedures at the same time (anogenital examination, photographs and disclosure). The conclusion of the study was that child sexual abuse investigations may be accomplished without undue psychophysiological stress when recommended precautions are taken. PMID- 16371261 TI - A tiered healthcare system for persons in police custody--the use of a forensic nursing service. AB - In February 1992 the healthcare programme for persons held in police custody within the Melbourne metropolitan area was restructured. This was undertaken as a joint exercise between Police, the Health Department and the Office of Corrections as a response to the need to develop an efficient and cost effective healthcare delivery system. A tiered medical response was instituted, utilising a forensic nurse for the primary medical contact with forensic physicians providing medical consultation and support as necessary. Criteria were developed regarding the delineation of duties appropriate for the forensic nurse and for the forensic physician. The nurses were provided with transport, communication and basic medical supplies enabling a flexible response to requests for medical attention and for the nurse to perform triage, a basic medical assessment and administer appropriate treatment. Over the 15-month period since inception over 3000 consultations have been performed by the forensic nurses. The service has been commended by police, doctors working in the prison system, forensic physicians and community groups. A comprehensive review has demonstrated the efficiency and reliability of such a service. Costs in providing the service have not increased. We conclude that a forensic nursing service is of considerable benefit in complementing a range of services provided by forensic medical officers in a metropolitan setting. PMID- 16371262 TI - Development of a clinical forensic medicine curriculum for emergency physicians in the USA. AB - To address the forensic needs of living patients, the Department of Emergency Medicine at the University of Louisville School of Medicine in Louisville, Kentucky, USA initiated the first clinical forensic medicine training programme in the USA. In July 1991, formal training in clinical forensic medicine was incorporated into the core curriculum of the USA's second oldest academic emergency medicine training programme. The University of Louisville, in cooperation with the Kentucky Medical Examiner's Office, developed the curriculum to provide the emergency physician with the knowledge base and technical skills to perform forensic evaluations of living patients. Forensic lectures are given monthly by local and regional forensic experts including: forensic pathologists, prosecuting attorneys, firearm and ballistics examiners, law enforcement officers, forensic chemists and forensic odontologists. Topics which are presented include: forensic pathology, forensic photography, ballistics and firearms analysis, paediatric physical and sexual assault, crime scene investigation, forensic odontology, courtroom and expert testimony and the forensic evaluation of penetrating trauma. As a result of the introduction of clinical forensic medicine into the core curriculum of an emergency medicine training programme the residents are now actively addressing the forensic issues encountered in the Emergency department. Key, often short-lived forensic evidence, which was frequently overlooked or discarded while delivering patient care is now recognized, documented and preserved. The development and introduction of a clinical forensic medicine curriculum into emergency medicine training has greatly enhanced the emergency physician's ability to recognize, document and address the forensic needs of their patients who are victims of violent and non-fatal trauma. PMID- 16371263 TI - Failure to thrive: a case obvious to physicians is perplexing to the court. AB - Proving a failure to thrive case in court can be difficult unless all professionals prepare in advance for consistent testimony which educates the court in a simple and straightforward manner. This case highlights the importance of a physician being alert to non-organic failure to thrive, as a cause of abuse or neglect. A bizarre array of confusing biological and psychosocial facts may prove difficult to categorise for forensic purposes. Physicians can themselves complicate future court presentations by failing to use growth charts to evaluate children and using non-speaking phrases such as 'well-nourished'. Although non organic failure to thrive can often be determined medically; attorneys, police and other healthcare personnel may benefit from the review of physical, psychosocial, and familial characteristics of failure to thrive children. A multi disciplinary team approach best serves the interests of these abused children as well as the interests of justice. PMID- 16371264 TI - Drug concealment in custody deaths--two cases. AB - Internal concealment of drugs of abuse is a well established method of transporting such substances to avoid detection while they are being smuggled across international frontier lines.(1,2) The same method has been used by dealers and 'pushers' in between purchases on their selling rounds to avoid detection by the police. These methods of hiding drugs may be used by persons taken into short-term custody, in their belief that potential withdrawal effects would thus be avoided by continued availability of their drugs. Two cases are reported in which such concealment was discovered at autopsy after death had occurred while in police custody. In one case the drug concealed was dihydrocodeine and was directly responsible for the death. PMID- 16371265 TI - Occupational health in police work: a Canadian perspective. PMID- 16371266 TI - Acute head injury: assessment and practical management. PMID- 16371267 TI - What now--what next? The future of clinical forensic medicine in the UK. PMID- 16371268 TI - The incidence of child homicide: how accurate are the official statistics? AB - This paper examines the incidence of child homicide in England and Wales, drawing on a review of files from the Director of Public Prosecutions in London from 1983 and 1984. After outlining the official statistics on child homicide, five factors are discussed which suggest that many cases of child-killing may be wrongly categorised. These factors are: cases where maltreatment is not the immediate cause of death, 'cot deaths', the legal difficulties of proof, cases where no body is found or identified and professional reluctance to act. Attempts to estimate the true incidence of child homicide are reviewed and the estimate calculated in the present study outlined. PMID- 16371269 TI - Suicide in custody. AB - Current knowledge about suicide in the inmates of jails and prisons is revieved. The strategies for preventing inmate suicide, including screening for suicidality, suicide-proofing the institution and establishing sound organisational policies and procedures, are examined. Alternatives to incarceration are also urged for those inmates who might fit well into non custodial settings. PMID- 16371270 TI - Autoerotic death--characteristic features and diagnostic difficulties. AB - Autoerotic death refers to the accidental death of individuals of either sex due to failure of devices, or unexpected effect of materials, that were being used to enhance the sexual experience. Although asphyxia from hanging has been described most frequently, a wide variety of other lethal situations have been reported. As there is great variability in the number and types of sexual aids that may be used by individuals, careful death scene examination is essential in helping to exclude suicide and more rarely homicide. The clandestine nature of this paraphilia may, however, make subsequent identification and diagnosis difficult unless the death scene features are typical. This is particularly so in the rare cases involving women. In the following paper the typical features of this disorder are reviewed with a description of less obvious cases and possible diagnostic pitfalls. PMID- 16371271 TI - An analysis of the factors implicated in fire deaths in children. AB - The aim of this study was to determine: 1) the factors implicated in fire deaths in children, 2) the relevance of blood carbon monoxide (CO) and hydrogen cyanide (CN) concentrations in child fire victims. 34 child fire victims (17 males: 17 females) were studied. Mean age was 4.5 years. In 12 cases medico-legal autopsies were performed, but no toxicological analyses were done (group A). In 14 cases autopsies were not performed, but blood CO and CN concentrations were measured in the victims (group B). In the remaining 8 cases both toxicological analyses and medicolegal autopsies were done (group C). In 29 cases out of 34, children had been left alone by their parents. In 20 cases out of 34, children had 80% burns of their body surface. In groups A and C autopsies enabled violence or other criminal cause of deaths to be excluded. All victims had smoke inhalation. The mean blood CN concentration in the 22 fire victims (groups B and C) was 70.67 +/- 14.11 micromol/l (0-207). The mean blood CO concentration was 1.89 +/- 0.51 mmol/l (0.11-5.89). There was no significant correlation between CN and the CO blood concentrations. There was no significant correlation between the burn surface area and the blood concentration of either CN or CO. In 11 cases, one or both gases concentrations were in the potential toxic range (CN: 40-<100 micromol/l; CO: 1-<5.8 micromol/l) but below the lethal range. Therefore these deaths were considered to be due to the combined effect of gas toxicity. It is concluded that: 1) In case of fire death, a medico-legal autopsy should be performed to confirm that victims suffered smoke inhalation and to exclude violence or other criminal cause of deaths; 2) Blood CN and CO measurements identify the exact contribution of each to the death; 3) Immediate blood sampling at the scene of death should be done because CN may disappear rapidly from body tissue after death. PMID- 16371272 TI - The extent of injecting and syringe sharing in prison reported by Edinburgh drug clinic attenders. AB - 68 outpatient attenders at Edinburgh's Community Drug Problem Service who reported a history of drug injecting and who had spent 1 week or more in prison between March 1987-March 1990 were interviewed about drug injecting behaviour and syringe sharing before and during each prison episode they reported. One-third of those injecting prior to prison continued to do so in prison, most doing so twice weekly or more. 80% of prison injectors shared syringes in prison. No subjects commenced their injecting career in prison, and being imprisoned did not cause past injectors who had stopped to resume injecting in custody. Being imprisoned tended to cause non-sharing injectors to resume sharing in prison. Prison injectors tended to have started both their oral and their injecting drug use careers at an earlier age, and to have been sharing syringes in the community prior to custody. Half of prison sharers used unofficially acquired bleach or disinfectant to sterilise injecting equipment. Continuing involvement with the courts and thus risk of reimprisonment was common. The findings support the case for harm reduction policies in Scottish prisons, and diversion from custody and into treatment for this high risk group. PMID- 16371273 TI - Sudden infant death syndrome in infants of cocaine using mothers. AB - A study of Sudden Infant Death Syndrome (SIDS) with maternal cocaine exposure was undertaken using the perinatal medical records at the University of Miami/Jackson Memorial Medical Center and case files of the Metropolitan Dade County Medical Examiner Department, Miami, Florida USA from 1988-1992. 78 SIDS cases were analysed. 18 infants were classified as cocaine-exposed SIDS infants with positive history of maternal cocaine abuse and/or positive urine toxicology; 50 infants were classified as cocaine-negative SIDS infants without the history and with negative toxicology. The incidence of SIDS among cocaine-exposed infants was estimated to be higher than that of cocaine-negative infants overall (p < 0.025), but statistical significance between the groups was not confirmed when controlled for ethnicity. No significant differences of birth weight, estimated gestational age, body and brain weights were observed at autopsy between the two groups. Cocaine-exposed SIDS cases included 6 infants born small for gestational age (p < 0.05). 10 of 18 cocaine-exposed SIDS infants had a positive result for urine toxicology at birth and died younger aged than cocaine-negative infants (p < 0.05). Toxicology tests of the specimens obtained from cadavers were all negative for cocaine metabolites. PMID- 16371274 TI - Prevalence of HIV risk factors for individuals examined in clinical forensic medicine. AB - OBJECTIVE: to identify the prevalence of high-risk factors for infection with Human Immunodeficiency Virus (HIV) in individuals examined in clinical forensic medical practice and to determine opinions and attitudes about HIV in this patient group. DESIGN: Anonymised questionnaire completed by consecutive individuals seen in clinical forensic medical practice. SETTING: Police stations in London attended by Group IV forensic medical examiners. SUBJECTS: 518 individuals examined in police stations (including prisoners and suspects, those detained in police custody, police officers and victims of crime). RESULTS: 164 (31.7%) individuals did not respond to the questionnaire because of: 1) refusal (12.6%) 2) inability because of drugs and/or alcohol (11.4%) 3) mental illness/disorder (4.2%) or 4) language difficulties (3.5%). 28.4% of the respondents were in at least one of the 'high-risk' categories for HIV infection. 26.5% were intravenous drug misusers; 15% were prostitutes; 9.8% (or their sexual partners) had lived in Central or East Africa since 1977; 5.9% were male homosexuals and 0.5% were haemophiliacs. 5.1% were infected with HIV or had Acquired Immunodeficiency Syndrome (AIDS). Only 28.8% of individuals always used condoms in short-term sexual relationships. 44.4% of respondents believed that everyone should be tested for HIV. CONCLUSION: Over one-quarter of the respondents were in higher-risk groups for infection with HIV. Almost one-third could or would not respond. It is concluded that it is not possible to identify by questionnaire, individuals at higher risk of HIV infection in forensic medical practice. This reinforces the necessity of observing good clinical practice to reduce contamination risks in this work environment. It is clear that education about risks for HIV infection is still much needed. PMID- 16371276 TI - The South African prison system: medical services. PMID- 16371275 TI - The promise, problems and limitations of methadone: a clinical perspective. AB - The use, advantages, disadvantages and side-effects of methadone are described. Outcome studies are reviewed. It is suggested that there is no good scientific evidence that dispensing methadone to non-compliant patients is an effective intervention. This is supported by studies at the Royal Newcastle Hospital which show that the longer the non-compliance the greater the failure rate. The dangers of continuing to prescribe methadone to patients using other illegally obtained drugs are emphasised. The need to supervise urine collection is highlighted. Alternatives to methadone maintenance (including therapeutic communities, narcotic antagonists, drug-free counselling, needle exchange programmes and benign neglect) are discussed. PMID- 16371277 TI - Chainsaw deaths: accident, suicide or murder? AB - This case review describes two deaths involving chainsaws. The first case describes a suicide by chainsaw and how it was differentiated from murder. The second case describes an accidental death associated with this tool which occurred within a week of the suicide in Dumfriesshire, Scotland. PMID- 16371279 TI - Munchausen syndrome by proxy. AB - Munchausen Syndrome by Proxy has been recognised for almost two decades. This review considers the definition, clinical features, consequences (to both victim and perpetrator), diagnosis and outcome of the syndrome. PMID- 16371280 TI - Respiratory symptoms among forensic identification workers. PMID- 16371281 TI - Assault and homicide by chop blows: a series of 11 cases. AB - In a series of robberies, 11 elderly women living alone were attacked by an assailant (HS) by means of chop blows directed primarily at the throat, but also at the head, neck, and thorax. 5 died of their injuries. 1 died immediately after the assault. The other 4 victims survived for between 7-38 days. As a result of the injuries incurred, the women were confused, physically weakened and confined to bed, and eventually died from pneumonia. Their death was thus casually related to the violence sustained. Extensive haematomas of the scalp and cervical soft tissue were present. In 3 cases there were injuries to the laryngeal skeleton and adjacent soft tissue. In 2 cases there were conjunctival haemorrhages. The other 6 attacks were not fatal. The blows led to transient unconsciousness, which often resulted in a state of confusion, sometimes mimicking cerebrovascular accidents (CVA). Of other injuries, craniofacial haematomas were the most common. These cases are reviewed in this paper. PMID- 16371282 TI - The potential for the use of personal computers in clinical forensic medicine. AB - This paper describes the use of a personal computer in clinical forensic medicine. It shows how a database can be easily created, updated for each new case by unqualified personnel, and then used to provide statistical data with a minimum resource input, without the need for great computer expertise. Databases will only recognise exactly identical abbreviations. Once created they can cross reference data and provide individually tailored statistics. Registration with the Data Protection Agency is required in the UK in order to hold personal information on file. The results of 2201 cases entered on the database of one forensic physician show high levels of drug and alcohol abuse co-existing with unemployment and homelessness. 53% of individuals brought to police stations as a place of safety (under Section 136 of the Mental Health Act 1983) were in need of admission to a psychiatric hospital. 12% of those individuals were of no fixed abode. The potential for a national database linking forensic physicians on a national basis, pooling their data thereby creating a powerful statistical tool is clear. PMID- 16371283 TI - The admissibility of scientific evidence in psychiatric malpractice: junk science and the Daubert case. AB - The defence in malpractice cases has traditionally relied upon a commonly accepted body of knowledge. According to the American 'Frye rule', this knowledge could either have been accepted generally or by a 'respectable minority' of physicians. The US Supreme Court, however, has recently ruled in the Daubert case that conclusions not subject to peer review are acceptable in malpractice cases. The authors analyse the implications of the Daubert decision using the case-study method. Two alternative-scenarios of a hypothetical case are analysed. The potential effect of Daubert places the US psychiatrist-defendant in an untenable position. Either use or non-use of non-peer-reviewed studies in clinical practice could produce a finding of negligence. Furthermore, the responsibility to assess scientific acceptability in US courts has shifted from expert witnesses to judges who are usually without scientific training. PMID- 16371284 TI - Artefact in forensic medicine: non-missile penetrating injury. AB - Gunshot wounds are well characterised in forensic textbooks and atypical or unusual ballistic characteristics feature regularly in forensic literature. Forensic practitioners receive continuing education in recognising a variety of gunshot wounds which are created by bullet missiles. Awareness of a non-missile penetrating injury mimicking a gunshot wound is also relevant to forensic medical examiners or others who are likely to attend a scene of death. In this case, which involved self-stabbing with a metal nail, a homicidal firearm death investigation was instigated because of a misinterpretation by crime scene investigators. PMID- 16371285 TI - Legal notes. AB - Clinical forensic medicine is practised in a constantly changing legal and social environment, for the law develops in response to pressures from society, from interest groups, and from academic jurisprudence. Medico-legal evidence is given in that context, requiring doctors to understand, for instance, why counsel may interrupt with a 'Don't answer that, doctor' which is followed by legal debate and a ruling from the presiding judge on admissibility. Such rules of evidence vary from country to country, none having an absolute claim to superiority. The editor welcomes brief reports on significant cases or pieces of legislation affecting the gathering and presentation of evidence and how these may influence witnesses and the decisions of courts. Readers of the Journal of Clinical Forensic Medicine come from many jurisdictions; a broad base of experience is, therefore, available for contributors to draw on to the benefit of all. PMID- 16371286 TI - A role for nurses in police station? PMID- 16371287 TI - Can we improve contraceptive use? PMID- 16371288 TI - Should coagulation tests be used to determine which oral contraceptive users have an increased risk of thrombophlebitis? PMID- 16371289 TI - Fertility control in wildlife: humans as a model. AB - In a time of accelerated extinction rates and biodiversity loss, it may seem illogical to be concerned with methods limiting the reproduction of wild populations; however, there is an urgent need to inhibit the proliferation of a wide variety of species. The range of animals for which fertility control is desired makes the development of a single method impossible. The various reproductive strategies used by individual species, the desired outcome of contraceptive programs (reversible or irreversible; male or female directed) and our ignorance of the reproductive biology of many endangered species necessitate thorough species-specific investigations. As fertility control in humans is a reality and research into methods of fertility control is more advanced, humans serve as a model for developing contraceptive approaches for wild species. Population control by traditional methods (indirect or direct intervention by culling, poisoning, translocation, etc.) is increasingly unacceptable to the public, making human studies even more valuable for finding solutions to overabundant wild populations. This review compares and contrasts the range of contraceptive methods used in both wildlife and humans. PMID- 16371290 TI - Treating acne with oral contraceptives: use of lower doses. AB - Oral contraceptives (OCs) have been shown to effectively treat acne. Clinical trials of various doses of ethinyl estradiol (EE) combined with progestins such as levonorgestrel, desogestrel, norgestimate, gestodene, cyproterone acetate and drospirenone in monophasic, triphasic and combiphasic formulations used to treat acne in women are reviewed here. Open-label and comparative studies beginning in the 1980s were the first to demonstrate objective and subjective reductions in the incidence of acne, severity of existing acne and seborrhea. Placebo controlled trials have corroborated these findings with a trend toward effective acne treatment with declining doses of EE. Significant reductions in total, inflammatory and noninflammatory lesions compared with placebo have been demonstrated with an OC containing the low dose of 20 microg of EE. Collectively, these findings support the use of low-dose OCs for the treatment of acne. PMID- 16371291 TI - Prospective, open-label, noncomparative study to assess cycle control, safety and acceptability of a new oral contraceptive containing gestodene 60 microg and ethinylestradiol 15 microg (Minesse). AB - OBJECTIVE: A prospective, open-label, noncomparative, multicenter study was carried out in 163 women aged 18-39 (mean 25+/-5 years), who used an ultra-low dose oral contraceptive pill (OCP) containing gestodene (GTD) 60 mug/ethinylestradiol (EE) 15 mug for 6 months. The objective of the study was to evaluate the acceptability, safety, bleeding patterns and premenstrual symptomatology in these women. METHODS: Patients used the OCP from Days 1-24, followed by a 4-day pill-free interval from Days 25-28 of the menstrual cycle. Physical and gynecological examinations were carried out at baseline and after 3 and 6 months, at which time blood pressure, weight, hemoglobin, hematocrit, SGOT, SGPT and urinalysis were also assessed. The Moos Menstrual Distress Questionnaire (MDQ) was completed on three consecutive days (Days 25-27 of the cycle) at baseline and at the end of the third and sixth cycles. Patients kept a menstrual diary throughout the study. RESULTS: A total of 146 women completed the study. Ten women discontinued because of adverse events and one undesired pregnancy occurred during treatment. No adverse metabolic effects were observed. The adverse event most frequently reported was breakthrough bleeding, which diminished, however, as the time of OC use increased. Cycle length and duration of bleeding decreased significantly with OC use (p<.01 and p<.05, respectively, after 6 months). Intensity of menstrual bleeding tended to decrease with OC use, but this difference was not statistically significant. Systolic and diastolic blood pressure were significantly lower after 6 months of OC use compared to baseline (p<.02). No alterations were recorded in body weight or laboratory evaluations. Statistically significant changes were found both in the total MDQ score and in several of the factors evaluated, and patients showed a statistically significant improvement in well-being with respect to premenstrual complaints and symptoms. CONCLUSION: This OC regimen is safe, well-accepted and well-tolerated, affects menstrual patterns beneficially by reducing both the intensity and duration of bleeding, provides good cycle control and improves premenstrual symptomatology. PMID- 16371292 TI - The use of an oral contraceptive containing ethinylestradiol and drospirenone in an extended regimen over 126 days. AB - OBJECTIVE: To assess the bleeding profile, acceptance and safety of an extended 126-day regimen of the oral contraceptive Yasmin [30 microg ethinylestradiol (EE) and 3 mg drospirenone (DRSP)]. METHODS: Using daily diaries, 177 women recorded bleeding events throughout the 126-day cycle. At end of treatment, the women completed questionnaires reflecting their satisfaction with the extended regimen. A subset of 30 women underwent endometrial histology sampling after completion of the extended regimen. RESULTS: Of 177 women assigned to the extended regimen, 80.8% completed the extended 126-day regimen. Approximately 40% of the women reported complete absence of bleeding, while in 60% a shift towards less intense bleeding was observed. The first onset of bleeding occurred after a median of 99.0 days into the extended cycle. The acceptance of the extended regimen was high, with 68.4% of the women expressing satisfaction. The general safety profile with the extended use was comparable to that seen with the conventional 21+7-day regimen. All endometrial biopsies with sufficient material for analysis were normal and supported the endometrial safety of the extended regimen. CONCLUSION: This study showed that the continuous use of a 30-microg EE and 3-mg DRSP formulation over 126 days was safe, efficacious, well accepted by the users and resulted in a considerable reduction of bleeding. PMID- 16371293 TI - Attitudes and prescribing preferences of health care professionals in the United States regarding use of extended-cycle oral contraceptives. AB - OBJECTIVE: The aim of this study was to examine the attitudes and prescribing patterns of health care professionals (HCPs) in the United States regarding extended continuous oral contraceptive (OC) regimens. DESIGN: A prospective survey of HCPs prescribing contraception who attended six educational conferences from April to July 2004 was conducted. METHODS: A survey instrument that included demographic variables, attitudes and prescribing preferences regarding extended OC regimens was developed. Surveys were administered at medical meetings to a diverse array of HCPs who specifically prescribe contraception in their practice. The survey was conducted immediately prior to a presentation at the medical meetings on contraceptive update. Returned forms were scanned into Excel databases and examined with Pearson's chi(2) and multivariate logistic regression methods to assess practices and attitudes. RESULTS: There were 551 HCPs, including primary care physicians, obstetrician/gynecologists, nurse practitioners and physician assistants, prescribing contraception who were surveyed at the medical meetings. Most of the participants (87.4%) thought that extended OC regimens should be routinely offered, with 81% of the respondents reporting that they had used extended regimens in their practice. Physicians and nurse practitioners who listed their specialty as "OB/GYN" were more likely to favor the concept of routinely offering the option of extended regimens (p<.0002) and to have recommended extended regimens in their practice (OR = 2.9, 95% CI = 1.8-4.6). Only 12% of the respondents thought that withdrawal bleeding with standard 21-day/7-day OC regimens "has health benefits and is necessary." The most commonly recommended extended regimen was an 84-day active pill use period followed by a 7-day hormone-free interval. CONCLUSION: Most HCPs prescribing contraception who attended the six medical meetings favored the use of extended OC regimens and have recommended them in their practice, especially if their specialty was OB/GYN. The most commonly prescribed extended regimen was an 84-day active pill use period followed by a 7-day hormone-free interval. PMID- 16371294 TI - Feasibility study of Nestorone-ethinylestradiol vaginal contraceptive ring for emergency contraception. AB - OBJECTIVE: The Nestorone/ethinylestradiol (NES/EE) vaginal ring is being developed as a regular contraceptive method by the Population Council. This ring is designed to release NES 150 microg/day and EE 15 microg/day during 1 year. Here, we report a Phase I clinical trial to determine the usefulness of this ring for emergency contraception. To that end, we tested the ability of this ring to interfere with ovulation when it is inserted during the follicular phase. METHOD: Forty-eight women protected from the risk of pregnancy by nonhormonal methods were divided into three groups, which differed by the size of the dominant follicle at the time of ring insertion: 12-14 mm (n = 16), 15-17 mm (n = 18) and >or=18 mm (n = 14) diameter. The NES/EE ring was left in the vagina for 7 consecutive days, after which it was removed. The growth of the leading follicle and plasma levels of estradiol, progesterone (P), luteinizing hormone (LH) and follicle stimulating hormone (FSH) in the ensuing 5 days after ring insertion were determined. Afterwards, steroid hormones were measured twice a week, until menses took place. All women had a control cycle before the ring cycle, and the range of maximum follicular diameter assigned to each volunteer was the same for the control and the ring cycle at the time when placebo was ingested or the ring inserted. RESULTS: During the 5-day period after ring insertion with follicles 12 17 mm, ovulation was absent in 25 of 34 cycles (p < .01 vs. control), and ovulatory dysfunction (absent, blunted or mistimed LH peak) occurred in 8 of the 9 remaining cycles (33/34 ovulatory processes altered; p < .005 vs. control). After ring insertion with follicles >or=18 mm in diameter, ovulation did not occur in 2 of 14 cycles or was dysfunctional in 7 of the 12 remaining cycles (9/14 ovulatory processes altered; p<.025 vs. control). Altogether, 87.5% of ring cycles (42/48) had either no ovulation or ovulatory dysfunction in the 5-day study period, in contrast to 39.6% (19/48 cycles) in control cycles (p < .001). Among follicles that failed to rupture within the 5-day study period, none ruptured later on in the ring-treated cycles, while 9 of 16 did so in control cycles. Sixty-two percent of ring-treated cycles were shorter than 24 days. Nausea, vaginal discharge and abdominal pain were the most frequently reported adverse events during ring use. CONCLUSION: Interference with 87.5% of ovulatory processes, without ovulation occurring later in the cycle and shortening of cycle length, suggests the NES/EE ring may be used as an emergency contraceptive method, with the potential advantage of providing continuing contraception after it has performed its emergency function. PMID- 16371295 TI - Self-injection of monthly combined hormonal contraceptive. AB - OBJECTIVES: The monthly injectable contraceptive is usually administered in the office. We aimed to compare home self-injection with office administration with respect to satisfaction, compliance, and time and money spent on contraceptive behavior. METHODS: We enrolled 16 subjects and taught them self-injection for a prospective cohort trial with crossover. Subjects performed three self-injections at home and then had three office injections by the nurse. RESULTS: Ten subjects completed the protocol. They reported similarly low pain and anxiety with nurse injections and self-injections. Women reported spending less money and similar time for home injections. Most subjects strongly preferred giving themselves injections at home to office injections and would recommend self-injection to other women. CONCLUSIONS: Self-injection of the monthly contraceptive at home is a preferable alternative to office administration for these subjects. This study demonstrates the feasibility of teaching women self-injection. Future studies should look at how best to teach women self-injection and to select women who will be successful with self-injection of contraceptives. PMID- 16371296 TI - Effects of a subdermal levonorgestrel contraceptive implant (Norplant) on serum cholesterol, triglycerides, ALT and AST in Iranian women. AB - BACKGROUND: To evaluate the effects of Norplant (36 mg of levonorgestrel, six capsules) on serum cholesterol, triglycerides, alanine transaminase (ALT) and aspartate transminase (AST), we enrolled 465 healthy women from Zahedan, Iran, into a longitudinal study. MATERIALS AND METHODS: Blood samples were collected after an overnight fast before implant insertion and after 3, 6, 9 and 12 months of use. RESULTS: Total cholesterol and triglyceride levels did not significantly change during Norplant use. Although there were statistically significant increases in ALT and AST levels during Norplant use, the values were within the reference range. PMID- 16371297 TI - FemCap with removal strap: ease of removal, safety and acceptability. AB - OBJECTIVES: FemCap is a silicone-rubber barrier contraceptive approved for marketing in the United States. To address reported problems with removal and dislodgment, the device's developer added a strap to the device and increased the height of the brim. This trial assessed whether the new design reduced removal difficulties and had any effects on dislodgment, genital pain/discomfort, safety, and acceptability. METHODS: Women used the strapped device for 8 weeks with follow-up visits at 2 and 8 weeks. Outcome measures were obtained through diary cards, questionnaires, and naked eye examination. Data from these 120 women were compared with data from 419 women who used the unstrapped FemCap in a previous contraceptive effectiveness study. RESULTS: The strapped device was not significantly easier for users to remove than the unstrapped device. Similar odds of dislodgment and cervical/vaginal irritation were seen with the two devices. Both female and male participants were significantly more likely to report pain/discomfort with use of the strapped device. Female users of the strapped device were significantly more likely to say they disliked their device. In six weeks, two pregnancies were observed, but pregnancy was not an endpoint in the study and no conclusions should be drawn regarding pregnancy rates. CONCLUSIONS: The modifications to the FemCap did not significantly improve the ease of device removal and appears to have resulted in significantly more female and male partner pain/discomfort and decreased acceptability, compared with the unstrapped device. PMID- 16371298 TI - Acceptability of a home monitor used to aid in conception: psychosocial factors and couple dynamics. AB - BACKGROUND: Assessing the psychological acceptability of technologies designed to assist couples in achieving pregnancy is complex. OBJECTIVE: The current study developed measures relating to the impact of one such technology on 52 couples' relationships, their feelings relating to pregnancy status and their feelings about the technology itself. METHODS: Pregnancy status and daily logs of sexual activity were recorded for four menstrual cycles, in addition to the completion of acceptability questionnaires. RESULTS: Baseline acceptability measures were more favorable among couples eventually achieving pregnancy. For couples not becoming pregnant, acceptability declined over time and relationships became more strained. Behavioral data clearly indicated a "targeting" and focusing of sexual activity in response to the information displayed by the monitor. CONCLUSION: Expectations of success, couple disagreements about prior failure and partner communication patterns appear to be related to pregnancy success when using such technology. PMID- 16371299 TI - Evaluating the clinical safety of three vaginal applicators: a pilot study conducted in the Dominican Republic. AB - BACKGROUND: Targeted research on applicator safety has not been conducted as part of microbicide clinical trials and was considered necessary for ensuring safe and effective product use. Colposcopy, regarded as a standard method for assessing the safety of vaginal products, including microbicides, was used in this study to assess the external genitalia, vaginal epithelium and cervical epithelium after a single applicator use. OBJECTIVES: The objectives of this study were to assess and compare the effects of three vaginal applicators on symptoms and signs of irritation of the external genitalia, vagina and cervix as seen by colposcopy after applicator use. METHODS: Twenty women used three different vaginal applicators over three separate clinic visits 7-14 days apart and underwent a colposcopic examination before and after each applicator use. In total, the safety of each applicator was assessed over 20 product uses. RESULTS: No severe colposcopic finding was reported. Four minor colposcopic findings (petechiae) were considered possibly related to product use. No difference was found between applicators. CONCLUSIONS: This study provides reassuring data on the safety of the three applicator products from the perspective of causing vaginal trauma or irritation. Epithelial changes that could be more important for risk of disease transmission were not observed. PMID- 16371300 TI - Lactobacillus species isolated from the vagina: identification, hydrogen peroxide production and nonoxynol-9 resistance. AB - BACKGROUND: Lactic acid production is considered to be the major protection mechanism of lactobacilli against vaginal infections due to genital pathogens. Some species of Lactobacillus are also hydrogen peroxide (H(2)O(2)) producers. Nonoxynol-9 (N-9) is a nonionic detergent and is the active component of many spermicidal preparations. It immobilizes sperm by disrupting the cell membrane and is believed to act similarly on a number of bacteria and viruses. It is known that N-9 inhibits Lactobacilli in vitro at concentrations of 0.1% to 1%. PURPOSE: The present study was conducted to identify the species of Lactobacillus isolated from vaginal fluids of reproductive-age women and to characterize the H(2)O(2) producing and N-9-resistant strains in an Argentine population. RESULTS: We identified Lactobacillus acidophilus, L. fermentum, L. gasseri, L. jensenii, L. casei subsp. casei, L. brevis and L. delbrueckii subsp. delbrueckii as the most frequent species. In this Argentine, South American population, 62% of women had H(2)O(2)-producing vaginal lactobacilli. We found a high number of sensitive strains. Sixty-two H(2)O(2)-producer strains were detected, 50 (80.6%) strains were sensitive to N-9 and 12 (19.4%) strains were resistant to the inhibitory effect of N-9. DISCUSSION: The vaginal microecologic findings are comparable to those found in other populations and suggest that (1) vaginal microecologic conditions are likely to be similar among Argentine women as in other countries, and (2) N-9 may have deleterious effects as in other populations. PMID- 16371301 TI - The optimal analysis of MRI data to quantify the distribution of a microbicide. AB - OBJECTIVE: The objective of this study was to systematically review the use of MRI for the evaluation of deployment characteristics of vaginal microbicides and to understand the relationship of gel spread with potential influencing factors. METHODS: Data from four clinical trials that used MRI to assess the deployment of a vaginal gel were combined. A linear mixed model best represented the spread of gel over time. Significant covariates that influence vaginal gel spread are baseline dimensions of the vagina, time from insertion, gel type, ambulation and volume of gel. RESULTS: These data demonstrate that MRI has outstanding intraperson validity and reproducibility. Therefore, paired design, using linear modeling adjusting for significant covariates, is the most efficient study design for comparison of products or volumes. Division of the vagina into two distinct anatomical regions best explains difference in gel spread, i.e., upper area (above the pelvic diaphragm) and lower area (below pelvic diaphragm). CONCLUSION: We conclude that the concept of spread from the cervix to the introits, in one dimension, is inadequate to explain spread of gel due to the complex shape of the vagina. PMID- 16371302 TI - Changes in menstrual pattern and ovarian function following bipolar electrocauterization of the fallopian tubes for voluntary surgical contraception. AB - OBJECTIVE: Our aim was to investigate the changes in menstrual pattern, ovarian reserve and presence of dysmenorrhea and ovulation after tubal ligation via bipolar electrocautery. METHODS: Sixty patients requesting voluntary tubal ligation were recruited in the study. Laparoscopic tubal sterilization via bipolar electrocoagulation was performed in all patients in the early follicular phase. Blood samples were collected on day 3, one cycle before the procedure, in the same cycle when the procedure was carried out and on the third cycle following the procedure for determination of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E(2)), and on day 21 for progesterone (P) levels. All patients were followed for 3 months, and changes in menstrual pattern, presence or absence of dysmenorrhea and ovulation were noted. RESULTS: Menstrual changes occurred in six patients (10%), although only one patient had mild dysmenorrhea (1.6%) after the procedure. The incidence of ovulation was 33% preoperatively, rising to 40% in the cycle when surgery was performed and maintained a constant level at 40% 3 months after tubal ligation. There was no statistically significant difference in the serum FSH, LH and estradiol levels in preoperative and postoperative assessments (p > .05). CONCLUSION: Tubal ligation has been blamed for causing luteal phase defect as a result of an effect on ovarian circulation. In our study, the rate of ovulation was slightly improved after the procedure, and ovarian reserve was not negatively affected. Bipolar electrocoagulation of the fallopian tubes did not alter the ovarian reserve and function. PMID- 16371303 TI - Effects of fenvalerate exposure on semen quality among occupational workers. AB - PURPOSE: The purpose of the study was to observe the effects of fenvalerate exposure on the semen quality of occupational workers. MATERIALS AND METHODS: Thirty-two male workers who were exposed to fenvalerate and 46 male administrators in the office in the same pesticide factory were selected as the exposure group and internal control group, respectively, and 22 male administrators in a center for disease control served as the external control group. In order to evaluate the exposure levels, the concentration of fenvalerate, toluene and xylene in the ambient air of the work place in these three groups were monitored simultaneously for 3 consecutive days. Moreover, the amount of fenvalerate in individual sampling and dermal contamination were evaluated in the exposure group and external control group. After the semen was collected according to the standard method, the workers' semen qualities were analyzed. RESULTS: Concentration of fenvalerate in the exposure areas was 21.55 x 10(-4) mg/m(3). The fenvalerate concentration in individual samplings in the exposure areas was 0.11 mg/m(3). The dermal contamination for workers in the fenvalerate exposure area was 0.05 mg/m(3). Fenvalerate was not detected in individual samplings collected in external areas. Sperm motion parameters through routine semen analysis in the exposure group were decreased significantly, and the abnormality rate of viscidity and coagulation was increased significantly as compared with the internal and the external control groups (p < .05 or p < .01). Furthermore, sperm progression and beat cross frequency (BCF) (4.20 +/- 1.68 Hz) in the exposure group were also significantly lower than those in the external control group by computer-assisted sperm motility analysis (CASA) (p < .05). CONCLUSION: Occupational exposure to fenvalerate could affect the semen quality of the workers, but the conclusion warrants further complete investigation due to various limitations of the study. PMID- 16371304 TI - Doses of levonorgestrel comparable to that delivered by the levonorgestrel releasing intrauterine system can modify the in vitro expression of zona binding sites of human spermatozoa. AB - BACKGROUND: The levonorgestrel-releasing intrauterine system (LNG-IUS) exerts its contraceptive effect by interfering with sperm transport through the female genital tract and with ovulation. However, the possibility cannot be discarded that the device exerts a direct effect on sperm function, thus, helping prevent fertilization. OBJECTIVES: The purpose of this study is to evaluate whether LNG at doses comparable to that measured in the uterus during the use of the LNG-IUS affects the detection of D-mannose binding sites or zona pellucida (ZP) receptors on human spermatozoa. The association with acrosomal status was also investigated. METHODS: Seventeen semen samples from fertile men were used, and spermatozoa were separated using a Percoll gradient and incubated for 22 h at 37 degrees C under 5% CO(2) in air. Capacitated spermatozoa were exposed for 30 min to 1,000 or 10,000 ng/mL of LNG or control medium. D-Mannose binding sites were detected using commercial D-mannosylated bovine serum albumin conjugated with fluorescein isothiocyanate, and the percentage of specific patterns (II and III) was recorded. The acrosome reaction was evaluated using the Pisum sativum technique. RESULTS: Levonorgestrel releasing significantly increased (p < .001) the percentage of spermatozoa with D-mannose receptors localized in pattern III, and this increase was dose dependent and a significant increase (p < .001) in the percentage of acrosome-reacted spermatozoa. Double staining confirmed an association between the location of the zona receptor and acrosomal status. RESULTS: The in vitro exposure of capacitated spermatozoa to the assayed doses of LNG increased the proportion of spermatozoa with fewer chances of interacting with the ZP. Further studies should be carried out to confirm whether this mechanism is part of the contraceptive action of the LNG-IUS. PMID- 16371305 TI - Effect of antispermatogenic agents on cell marker enzymes of rat Sertoli cells in vitro. AB - OBJECTIVE: To examine the role of Sertoli cells in the antispermatogenic action of two nonsteroidal male contraceptive compounds (CDRI-84/35 and gossypol) by evaluating their effect on some key parameters of Sertoli cell function in vitro. METHODS: Primary cultures of Sertoli cell were established from 18-day-old rat testis and treated on day 5 with different concentrations (1.0, 0.1, 0.01, and 0.001 mM) of either CDRI-84/35 or gossypol in vitro. Lactate (secretion), along with beta-glucuronidase, gamma-glutamyl transpeptidase, lactate dehydrogenase (LDH) and aromatase activities, was measured in these cells to examine the functions targeted by antispermatogenic agents in Sertoli cells. RESULTS: CDRI 84/35 significantly affected Sertoli cell parameters (stimulation in most of the cases) that are important for germ cell development like lactate secretion, LDH activity, aromatase activity (estradiol secretion) and so on. Gossypol in comparison to CDRI-84/35 had a more severe effect on Sertoli cells with complete inhibition of enzyme activities at higher concentrations. CONCLUSION: It is probable that the antispermatogenic action of CDRI-84/35 and gossypol is routed through Sertoli cells by disruption of important cell functions that support spermatogenesis in vivo. However, the two compounds appear to have different course of action in Sertoli cells, ultimately leading to spermatogenic failure. PMID- 16371306 TI - Effectiveness of LNG EC not fully explained by ovulatory dysfunction. PMID- 16371308 TI - Failed medical termination of twin pregnancy with mifepristone: a case report. PMID- 16371310 TI - Concerns with the report from Wildemeersch et al. PMID- 16371312 TI - Unintended pregnancies with Implanon. PMID- 16371314 TI - Methamphetamine-induced neuronal apoptosis involves the activation of multiple death pathways. Review. AB - The abuse of the illicit drug methamphetamine (METH) is a major concern because it can cause terminal degeneration and neuronal cell death in the brain. METH induced cell death occurs via processes that resemble apoptosis. In the present review, we discuss the role of various apoptotic events in the causation of METH induced neuronal apoptosis in vitro and in vivo. Studies using comprehensive approaches to gene expression profiling have allowed for the identification of several genes that are up-regulated or down-regulated after an apoptosis-inducing dose of the drug. Further experiments have also documented the fact that the drug can cause demise of striatal enkephalinergic neurons by cross-talks between mitochondria-, endoplasmic reticulum- and receptor-mediated apoptotic events. These neuropathological observations have also been reported in models of drug induced neuroplastic alterations used to mimic drug addiction (Nestler, 2001). PMID- 16371315 TI - Salmonella lipopolysaccharide (LPS) mediated neurodegeneration in hippocampal slice cultures. AB - Neuroinflammation has been suggested to play an integral role in the pathophysiology of various neurodegenerative diseases. Bacterial lipopolysaccharide (LPS) endotoxins are general activators of immune-cells, including microglial cells, which induce expression of pro-inflammatory factors. The aim of this study was to characterize neurodegenerative effects of exposure to LPS, derived from Salmonella abortus equi bacteria, in an in vitro brain slice culture system. Quasi-monolayer cultures were obtained using roller-drum incubations of hippocampal slices from neonatal Sprague Dawley rats for three weeks. Microglia/macrophages were identified in the monolayer cultures by CD11b immunostaining, while neuronal populations identified included N-methyl-D aspartate (NMDA-R1) receptor immunoreactive pyramidal neurons and smaller GABA immunoreactive cells. Following exposure to LPS (100 ng/ml) an increased density of CD11b positive cells was found in the cultures. In addition, the LPS exposure produced a concentration-dependent loss of the NMDA-R1 immunoreactive neurons in the cultures which was substantial at 100 ng/ml LPS. The loss of NMDA-R1 cells was apparent already after 24 h exposure to LPS and seemed to be primarily due to necrotic-like cell death. However, a continued loss of cells was found when cultures were analyzed at 72 h, concomitant with an increase in the expression of p53 in the NMDA-R1 cells and TUNEL labeling of a few cells. Also the number of GABA-immunoreactive cells decreased rapidly and to a substantial extent after 24 h exposure to LPS, with a continued decrease up to 72 h. The findings show that Salmonella LPS increases the density of CD11b positive cells and acts as a potent neurotoxin in hippocampal roller-drum slice cultures. The LPS-induced neurodegeneration has both necrotic- and apoptotic-like properties and appears to be non-selective, affecting both pyramidal and GABA neurons. LPS-induced neurotoxicity in slice cultures may be a useful system to study processes involved in inflammatory-mediated neurodegeneration. PMID- 16371316 TI - Role of astrocytes in glutamate homeostasis: implications for excitotoxicity. AB - Glutamate homeostasis in the brain is maintained by its well balanced release, uptake and metabolism. It appears that astrocytes play a prominent role in this context since they possess a very powerful battery of glutamate transporters. Thus, malfunction of astrocytic glutamate transporters will lead to an excessively high extracellular glutamate concentration which may result in neurodegeneration caused by the excitotoxic action of glutamate. PMID- 16371317 TI - Degree of damage compensation by various PACAP treatments in monosodium glutamate induced retinal degeneration. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) has been shown to be neuroprotective in retinal ischemia and monosodium L-glutamate (MSG)-induced retinal degeneration. Here we describe how different MSG treatments (1x and 3x application) cause retinal damage and finally lead to the destruction of the entire inner retina and how PACAP attenuates this effect. Newborn rats from both sexes were injected subcutaneously with 2 mg/g bodyweight MSG on postnatal days 1, 5 and 9. The left eye was left intact while we injected 5 microl PACAP38 solution (100 pmol) into the vitreous of the right eye with a Hamilton syringe at the time of (i) the first, (ii) the first two or (iii) all three MSG injections. Histological analysis has shown that the above described MSG treatment caused the entire inner plexiform layer (IPL) to degenerate, and the inner nuclear (INL) and ganglion cell layers (GCL) seemed fused. One time PACAP38 treatment at the first MSG application did not change the degenerative capacity of MSG. However, if animals received PACAP38 into the vitreous of the eye at the first 2 or all 3 times, a substantial protective effect could be observed. The IPL remained well discernible, the INL retained 2-3 cell rows and the number of cells in the GCL was substantially higher than in the MSG-treated retinas, and was not significantly different from that observed in the control tissue. We conclude that (i) 2 or 3 times PACAP treatment attenuates retinal degeneration; (ii) one PACAP treatment does not provide protection against repeated excitotoxic insults, and (iii) repeated application of PACAP under these experimental conditions may lead to a primed state in which further neurotoxic insults are ineffective. PMID- 16371318 TI - Development of neurological reflexes and motor coordination in rats neonatally treated with monosodium glutamate. AB - Monosodium glutamate (MSG) treatment of neonatal rats causes neuronal degeneration in various brain areas and leads to several neurochemical, endocrinological and behavioral alterations. However, relatively little is known about the development of neurological reflexes and motor coordination of these animals. Therefore, the aim of the present study was to examine the neurobehavioral development of newborn rats treated with MSG. Rats received MSG at postnatal days 1, 3, 5, 7, and 9. Appearance of neural reflexes and reflex performance as well as motor coordination were examined for 5 weeks after birth. The efficacy of MSG treatment was confirmed by histological examination of the arcuate nucleus. We found that MSG treatment delayed the appearance of forelimb placing, forelimb grasp and righting reflexes, besides the retarded somatic development. The treated pups performed surface righting in significantly longer times. Also, worse performance was observed in the foot-fault and rota-rod tests. However, MSG-treated rats reached control levels by the end of the fifth postnatal week. These results show that MSG treatment does not cause permanent alterations in the neurobehavioral development, only delays the appearance of some reflexes and leads to temporary changes in reflex performance and motor coordination signs. PMID- 16371319 TI - NMDA receptors as targets of heavy metal interaction and toxicity. AB - The N-methyl-D-aspartate (NMDA) receptor (NR) is a ligand-gated channel that carries the slow component of the glutamate-activated postsynaptic current. Divalent metal ions can affect the NR channel activity in a voltage-dependent [Mg(II)-like] or voltage-independent [Zn(II)-like] manner. We have studied the effect of two toxic metals, lead [Pb(II)] and nickel [Ni(II)] on recombinant NR1a NR2A and NR1a-NR2B channels expressed in RNA-injected Xenopus laevis oocytes or in transiently transfected mammalian HEK293 cells. Pb(II) caused a dose dependent, but voltage-independent reversible inhibition of NMDA-activated channel activity similar for NR2A and NR2B-containing receptors; it did not modify the single channel conductance, indicating that its binding site is located out of the ionic pathway of permeation. On the contrary, Ni(II) had multiple and complex effects on NR channels. It determined a voltage-dependent, Mg(II)-like block by which the single channel amplitude and the mean open time were reduced in both NR2A and NR2B-containing channels. While high (>100 microM) concentrations caused a dose-dependent reduction of the activity in both channel types, 30 microM determined a voltage-independent decrease in the frequency of NR1a-NR2A channel openings, but an increase in the frequency of NR1a-NR2B channel openings, confirming previous observations of a subunit-dependent effect of this metal. These results were interpreted under the hypothesis that Pb(II) mediates a Zn(II)-like voltage-independent allosteric modulation that, different from Zn(II), is subunit-independent. In contrast, Ni(II) has different modes of action, which are dependent on the NR2 subunit type present in the receptor and are likely to be related to different interaction sites. The NR2B-dependent facilitation bears close similarities with the polyamine-mediated potentiation. PMID- 16371320 TI - Behavioral alterations in Lewis rats following two-day continuous 3 nitropropionic acid administration. AB - The mitochondrial toxin, 3-nitropropionic acid (3-NP), produces motor dysfunction and striatal atrophy in rats. However, rat strain and method of administration may contribute to variability in the deficits caused by 3-NP toxicity. To evaluate this, changes in nocturnal spontaneous locomotor activity from chronic administration of 3-NP using an osmotic mini pump, were examined in the Lewis rats. Lewis rats were treated with 3-NP or saline for 2 days and behavior was tested daily for a 15 day period. Animals receiving 3-NP displayed significantly less spontaneous activity than animals in the saline group. 3-NP treated animals also weighed significantly less when compared to saline treated animals. These results demonstrate that even though there were no significant alterations in overt anatomical pathology, even short-term exposure to 3-NP produced significant effects. This short-term administration may present a potential paradigm for examination of sub-threshold neurotoxicity. PMID- 16371321 TI - Cytokine production by a human microglial cell line: effects of beta-amyloid and alpha-melanocyte-stimulating hormone. AB - Senile plaques in the Alzheimer's disease (AD) are formed by aggregation of beta amyloid (Abeta) peptide. Abeta peptide has been shown to activate microglia and stimulate their production of inflammatory factors, such as cytokines. In the AD brain, the continued presence of amyloid plaques may keep microglia persistently activated, leading to chronic inflammation in the CNS. It is well established that alpha-melanocyte-stimulating hormone (alpha-MSH) gives rise to anti inflammatory and anti-pyretic effects. The biological activities of alpha-MSH are mediated by one or more of the melanocortin receptor (MCR) subtypes, i.e. MCR1 - MCR5. The aim of the present study was to determine the effect of alpha-MSH alone and on Abeta-activated microglial cells with regard to the secretion of inflammatory cytokines, such as interleukin-6 (IL-6), and to determine which receptor subtype mediates the effects of alpha-MSH. The human microglial cell line, CHME3, was incubated for 24 h with freshly dissolved Abeta(1-40), interferon-gamma (IFN-gamma) and/or alpha-MSH. Freshly dissolved Abeta(1-40) (5 60 microM) resulted in a dose-dependent decrease in cell viability, along with a dose-dependent increase in IL-6 release. Neither IFN-gamma nor alpha-MSH affected the Abeta-induced secretion of IL-6, but resulted in a dose-dependent increase in basal IL-6 release. Agouti, the endogenous antagonist of MCR1 and 4, further increased the alpha-MSH-induced secretion of IL-6. RT-PCR showed the expression of MCR1, MCR3, MCR4 and MCR5 mRNA. The combined data suggest that the effect of alpha-MSH in increasing IL-6 release from the human microglial cell line is mediated by MCR3 or MCR5. PMID- 16371322 TI - Pro- and anti-inflammatory cytokines regulate the ERK pathway: implication of the timing for the activation of microglial cells. AB - Pro-inflammatory molecules induce glial activation and the release of potentially detrimental factors capable of generating oxidative damage, such as nitric oxide (NO) and superoxide anion (O2.-). Activated glial cells (astrocytes and microglia) are associated to the inflammatory process in neurodegenerative diseases. A strong inflammatory response could escape endogenous control becoming toxic to neurons and contributing to the course of the disease. We evaluated in a hippocampal cells-microglia co-culture model, if the pro-inflammatory condition induced by lipopolysaccharide + interferon-gamma (LPS+IFN-gamma) promoted damage directly or if damage was secondary to glial activation. In addition, we explored the effect of the anti-inflammatory cytokine transforming growth factor-beta1 (TGF-beta1), and pro-inflammatory cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) on the regulation of the inflammatory response of microglia. We found that LPS+IFN-gamma-induced damage on hippocampal cultures was dependent on the presence of microglial cells. In hippocampal cultures exposed to LPS+IFN-gamma, TGF-beta1 was induced whereas in microglial cell cultures LPS+IFN-gamma induced the secretion of IL-1beta. TGF-beta1 and IL 1beta but not TNF-alpha decreased the NO production by 70-90%. PD98059, an inhibitor of MAP kinase (MEK), reduced the IFN-gamma-induced NO production by 40%. TGF-beta and IL-1beta reduced the IFN-gamma induced phosphorylation of ERK1,2 by 60% and 40%, respectively. However, the effect of IL-1beta was observed at 30 min and that of TGF-beta1 only after 24 h of exposure. We propose that acting with different timing, TGF-beta1 and IL-1beta can modulate the extracellular signal-regulated kinase ERK1,2, as a common element for different transduction pathways, regulating the amplitude and duration of glial activation in response to LPS+IFN-gamma. Cross-talk among brain cells may be key for the understanding of inflammatory mechanisms involved in pathogenesis of neurodegenerative diseases. PMID- 16371323 TI - Neuroprotective effects of nicotine against salsolinol-induced cytotoxicity: implications for Parkinson's disease. AB - Parkinson's disease is associated with degeneration of dopaminergic cell bodies in the substantia nigra. It has been suggested that salsolinol, an endogenous metabolite of dopamine, may be involved in this process. An inverse relationship between Parkinson's disease and smoking (nicotine intake) has been observed in epidemiological studies. Moreover, neuroprotective effects of nicotine in various experimental models have been observed. In this study we sought to determine whether salsolinol-induced cytotoxicity in SH-SY5Y human neuroblastoma cells, a cloned cell line which expresses dopaminergic activity, could also be prevented by nicotine pretreatment, and if so, which nicotinic receptors may mediate the actions of nicotine. Exposure of SH-SY5Y cells to 0.8 mM salsolinol for 24 hours resulted in approximately 80% cell death as determined by 3,[4,5-dimethylthiazol 2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Pretreatment of cells with 0.1 mM nicotine resulted in inhibition of salsolinol-induced cytotoxicity. The effects of nicotine were blocked by mecamylamine, a non-selective nicotinic antagonist as well as conotoxins with selective antagonism against alpha3 containing nicotinic receptor subunits. The effects of nicotine were not affected by dihydro-beta-erythroidine or methyllycaconitine, selective antagonists against alpha4-beta2 or alpha7 nicotinic receptors, respectively. It is suggested that selective nicotinic agonists may be of therapeutic potential in at least a subpopulation of Parkinsonian patients. PMID- 16371325 TI - A review of childhood and adolescent obesity interventions. AB - Obesity is one of the largest health problems facing Americans. Figures produced by the National Center for Health Statistics show that in 1999-2002, 15.8% of 6 11-year-olds were overweight as were 16.1% of adolescents aged 12-19. Some have proposed that interventions in childhood may be the best place to focus efforts to reduce the growing obesity epidemic. This paper examines several of the popular intervention strategies that have been proposed for youth and assesses the potential of these interventions for meaningful public health impacts. Findings show that most childhood obesity interventions completed to date produced only meager results, creating need for more well-designed studies that are tailored to the needs of specific age, ethnic and economic groups. PMID- 16371324 TI - Role of the JAKs/STATs pathway in the intracellular calcium changes induced by interleukin-6 in hippocampal neurons. AB - Recent studies show that inflammation has an active role in the onset of neurodegenerative diseases. It is known that in response to extracellular insults microglia and/or astrocytes produce inflammatory agents. These contribute to the neuropathological events in the aging process and neuronal degeneration. Interleukin-6 (IL-6) has been involved in the pathogenesis of neurodegenerative disorders, such as Alzheimer's and Parkinson's diseases. Here, we show that IL-6 treatment of rat hippocampal neurons increases the calcium influx via NMDA receptor, an effect that is prevented by the specific NMDA receptor antagonist MK 801 (dizocilpine). We also show that this calcium influx is mediated by the JAKs/STATs pathway, since the inhibitor of JAKs/STATs pathway, JAK 3 inhibitor, blocks calcium influx even in the presence of IL-6. This increase in calcium signal was dependent on external sources, since this signal was not observed in the presence of EGTA. Additional studies indicate that the increase in cytosolic calcium induces tau protein hyperphosphorylation, as revealed by using specific antibodies against Alzheimer phosphoepitopes. This anomalous tau hyperphosphorylation was dependent on both the JAKs/STATs pathway and NMDA receptor. These results suggest that IL-6 would induce a cascade of molecular events that produce a calcium influx through NMDA receptors, mediated by the JAKs/STATs pathway, which subsequently modifies the tau hyperphosphorylation patterns. PMID- 16371326 TI - Does the mind reflect the mouth? Sensory profiling and the future. AB - Descriptive Sensory Profiling methods have been used for over half-a-century. As the application of these methods obviously does not occur in a vacuum, we sketch the contextual basis of these methods. A unifying model, illustrating the tasks required from a sensory panellist, is presented with the aim of illuminating some major differences between methods. The history and main developments in the field of descriptive sensory profiling are outlined in a basic pedigree of methods. The authors suggest an approach to sensory profiling where taylor-made, problem oriented, methods are devised and used. PMID- 16371327 TI - Caffeine use in sports, pharmacokinetics in man, and cellular mechanisms of action. AB - Caffeine is the most widely consumed psychoactive 'drug' in the world and probably one of the most commonly used stimulants in sports. This is not surprising, since it is one of the few ergogenic aids with documented efficiency and minimal side effects. Caffeine is rapidly and completely absorbed by the gastrointestinal tract and is readily distributed throughout all tissues of the body. Peak plasma concentrations after normal consumption are usually around 50 microM, and half-lives for elimination range between 2.5-10 h. The parent compound is extensively metabolized in the liver microsomes to more than 25 derivatives, while considerably less than 5% of the ingested dose is excreted unchanged in the urine. There is, however, considerable inter-individual variability in the handling of caffeine by the body, due to both environmental and genetic factors. Evidence from in vitro studies provides a wealth of different cellular actions that could potentially contribute to the observed effects of caffeine in humans in vivo. These include potentiation of muscle contractility via induction of sarcoplasmic reticulum calcium release, inhibition of phosphodiesterase isoenzymes and concomitant cyclic monophosphate accumulation, inhibition of glycogen phosphorylase enzymes in liver and muscle, non-selective adenosine receptor antagonism, stimulation of the cellular membrane sodium/potassium pump, impairment of phosphoinositide metabolism, as well as other, less thoroughly characterized actions. Not all, however, seem to account for the observed effects in vivo, although a variable degree of contribution cannot be readily discounted on the basis of experimental data. The most physiologically relevant mechanism of action is probably the blockade of adenosine receptors, but evidence suggests that, at least under certain conditions, other biochemical mechanisms may also be operational. PMID- 16371328 TI - Authenticity assessment of dairy products. AB - The authenticity of dairy products has become a focal point, attracting the attention of scientists, producers, consumers, and policymakers. Among many others, some of the practices not allowed in milk and milk products are the substitution of part of the fat or proteins, admixtures of milk of different species, additions of low-cost dairy products (mainly whey derivatives), or mislabeling of products protected by denomination of origin. A range of analytical methods to detect frauds have been developed, modified, and continually reassessed to be one step ahead of manufacturers who pursue these illegal activities. Traditional procedures to assess the authenticity of dairy products include chromatographic, electrophoretic, and immunoenzymatic methods. New approaches such as capillary electrophoresis, polymerase chain reaction, and isotope ratio mass spectrometry have also emerged alongside the latest developments in the former procedures. This work intends to provide an updated and extensive overview since 1991 on the principal applications of all these techniques together with their advantages and disadvantages for detecting the authenticity of dairy products. The scope and limits of different tools are also discussed. PMID- 16371329 TI - Liposomal nanocapsules in food science and agriculture. AB - Liposomes, spherical bilayer vesicles from dispersion of polar lipids in aqueous solvents, have been widely studied for their ability to act as drug delivery vehicles by shielding reactive or sensitive compounds prior to release. Liposome entrapment has been shown to stabilize encapsulated, bioactive materials against a range of environmental and chemical changes, including enzymatic and chemical modification, as well as buffering against extreme pH, temperature, and ionic strength changes. Liposomes have been especially useful to researchers in studies of various physiological processes as models of biological membranes in both eukaryotes and prokaryotes. Industrial applications include encapsulation of pharmaceuticals and therapeutics, cosmetics, anti-cancer and gene therapy drugs. In the food industry, liposomes have been used to deliver food flavors and nutrients and more recently have been investigated for their ability to incorporate food antimicrobials that could aid in the protection of food products against growth of spoilage and pathogenic microorganisms. In this review we briefly introduce key physicochemical properties of liposomes and review competing methods for liposome production. A survey of non-agricultural and food applications of liposomes are given. Finally, a detailed up-to-date summary of the emerging usage of liposomes in the food industry as delivery vehicles of nutrients, nutraceuticals, food additives, and food antimicrobials is provided. PMID- 16371330 TI - Nutraceutical applications of garlic and the intervention of biotechnology. AB - Garlic (Allium sativum L.) is an important and widely cultivated plant with both culinary and medicinal uses stemming from its biological activities, which include antibiotic, anticancer, anti-thrombotic, and lipid-lowering cardiovascular effects. Though such medicinal use of garlic existed for centuries, there was little scientific support for its therapeutic and pharmacological properties. However, there has been a recent upsurge of research on garlic aiming to understand its exact mechanism of action in each case so that garlic and its products may have more judicious future applications. Since garlic is vegetatively propagated, its improvement for desired traits through conventional means is difficult. The intervention of biotechnological methods such as tissue culture and gene transfer protocols developed recently hold great promise for improving this crop. Due to new innovations in instrumentation and processing technologies coupled with more judicious experimental models, better products are foreseen in the market. The objective of this article was to review the recent developments made towards understanding the mechanism by which garlic imparts different therapeutic effects as well as to review what biotechnology can offer to improve this crop and its products. PMID- 16371331 TI - Post harvest spoilage of sweetpotato in tropics and control measures. AB - Sweetpotato storage roots are subjected to several forms of post harvest spoilage in the tropical climate during transportation from farmers' field to market and in storage. These are due to mechanical injury, weight loss, sprouting, and pests and diseases. Sweetpotato weevil is the single most important storage pest in tropical regions for which no control measures or resistant variety are yet available. Several microorganisms (mostly fungi) have been found to induce spoilage in stored sweetpotatoes. The most important among them are Botryodiplodia theobromae, Ceratocystis fimbriata, Fusarium spp., and Rhizopus oryzae. The other less frequently occurring spoilage microorganisms include Cochliobolus lunatus (Curvularia lunata), Macrophomina phaseolina, Sclerotium rolfsii, Rhizoctonia solani, Plenodomus destruens. Microbial spoilage of sweetpotato is found associated with decrease in starch, total sugar, organic acid (ascorbic acid and oxalic acid) contents with concomitant increase in polyphenols, ethylene, and in some instances phytoalexins. Several methods are used to control microbial spoilage. Curing to promote wound healing is found as the most suitable method to control microbial spoilage. Curing naturally occurs in tropical climates where mean day temperature during sweetpotato harvesting season (February-April) invariably remains at 32-35 degrees C and relative humidity at 80-95%. Sweetpotato varieties varied in their root dry matter content, and low root dry matter content attributed for their high curing efficiency. Curing efficiency of varieties also differed in response to curing periods. Fungicide treatment, bio-control, gamma irradiation, hydro warming, and storage in sand and saw dust were found to have intermediate impacts in controlling spoilage and enhancing shelf life of sweetpotato roots. Breeding program has to be chalked out to develop new varieties suitable to curing under tropical conditions in addition to developing varieties having multi-spectrum resistance to major post harvest rot pathogens and sweetpotato weevils. PMID- 16371332 TI - Milk biologically active components as nutraceuticals: review. AB - Milk contains components that provide critical nutritive elements, immunological protection, and biologically active substances to both neonates and adults. Milk proteins are currently the main source of a range of biologically active peptides. Concentrates of these peptides are potential health-enhancing nutraceuticals for food and pharmaceutical applications. Several bioactive peptides may be used as nutraceuticals, for example, in the treatment of diarrhea, hypertension, thrombosis, dental diseases, as well as mineral malabsorption, and immunodeficiency. Minor whey proteins, such as lactoferrin, lactoperoxidase, lysozyme, and immunoglobulins, are considered antimicrobial proteins. Milk also contains some natural bioactive substances. These include oligosaccharides, fucosylated oligosaccharides, hormones, growth factors, mucin, gangliosides, and endogenous peptides, which are present in milk at secretion. Most of the claimed physiological properties of milk bioactive components have been carried out in vitro or in animal model systems, and these hypothesized properties remain to be proven in humans. Whether these milk bioactive components will replace drugs entirely in the immediate future is still unclear, but the increasing appreciation of "drug foods" or nutraceuticals plays a complementary rather than a substitutional role to the synthetic pharmacological drugs. PMID- 16371333 TI - Edible coatings for fresh-cut fruits. AB - The production of fresh-cut fruits is increasingly becoming an important task as consumers are more aware of the importance of healthy eating habits, and have less time for food preparation. A fresh-cut fruit is a fruit that has been physically altered from its original state (trimmed, peeled, washed and/or cut), but remains in a fresh state. Unfortunately since fruits have living tissue, they undergo enzymatic browning, texture decay, microbial contamination, and undesirable volatile production, highly reducing their shelf life if they are in any way wounded. Edible coatings can be used to help in the preservation of minimally processed fruits, providing a partial barrier to moisture, oxygen and carbon dioxide, improving mechanical handling properties, carrying additives, avoiding volatiles loss, and even contributing to the production of aroma volatiles. PMID- 16371334 TI - Lutein: a valuable ingredient of fruit and vegetables. AB - Lutein is a human serum carotenoid which is not synthesized by humans and thus must be obtained by the ingestion of food containing it such as fruits and vegetables. Lutein is present in different forms in those foods as all-trans lutein, cis-lutein, epoxi-lutein, and lutein linked to proteins. It discusses if the intake of lutein or diets supplemented with lutein or diets rich in fruits and vegetables are important in the prevention of diseases like some cancers, cardiovascular diseases, etc., that may be affected by the antioxidant effect of lutein; or in the prevention of age-related macular degeneration and other eye diseases. The concentration of lutein in fruits and vegetables depends on the species. We've included the concentration of lutein in 74 species reported by different authors since 1990. Currently the quantification of lutein is mainly performed by HPLC, but more investigations into a quantification method for lutein, lutein isomers, and epoxi-lutein are necessary. Improvement of lutein extraction methods is important as well. Methods commonly used in the vegetable and fruit industry like heat treatment, storage conditions, etc. can change lutein concentrations; other factors depend on the plant, for instance the variety, the stage of maturity, etc. PMID- 16371335 TI - Utilisation of a packed-bed biofilm reactor for the determination of the potential of biofilm accumulation in water systems. AB - An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm2 (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, "the biofilm accumulation potential", represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential. PMID- 16371336 TI - Flowing biofilms as a transport mechanism for biomass through porous media under laminar and turbulent conditions in a laboratory reactor system. AB - Fluid flow has been shown to be important in influencing biofilm morphology and causing biofilms to flow over surfaces in flow cell experiments. However, it is not known whether similar effects may occur in porous media. Generally, it is assumed that the primary transport mechanism for biomass in porous media is through convection, as suspended particulates (cells and flocs) carried by fluid flowing through the interstices. However, the flow of biofilms over the surfaces of soils and sediment particles, may represent an important flux of biomass, and subsequently affect both biological activity and permeability. Mixed species bacterial biofilms were grown in glass flow cells packed with 1 mm diameter glass beads, under laminar or turbulent flow (porous media Reynolds number = 20 and 200 respectively). The morphology and dynamic behavior reflected those of biofilms grown in the open flow cells. The laminar biofilm was relatively uniform and after 23 d had inundated the majority of the pore spaces. Under turbulent flow the biofilm accumulated primarily in protected regions at contact points between the beads and formed streamers that trailed from the leeward face. Both biofilms caused a 2 to 3-fold increase in friction factor and in both cases there were sudden reductions in friction factor followed by rapid recovery, suggesting periodic sloughing and regrowth events. Time-lapse microscopy revealed that under both laminar and turbulent conditions biofilms flowed over the surface of the porous media. In some instances ripple structures formed. The velocity of biofilm flow was on the order of 10 mum h(-1) in the turbulent flow cell and 1.0 mum h( 1) in the laminar flow cell. PMID- 16371337 TI - Larval development and post-settlement metamorphosis of the barnacle Balanus albicostatus Pilsbry and the serpulid polychaete Pomatoleios kraussii Baird: Impact of a commonly used antifouling biocide, Irgarol 1051. AB - AbstractThe impact of a commonly-used antifouling algicide, Irgarol 1051, on the larval development and post-settlement metamorphosis of the barnacle, Balanus albicostatus Pilsbry (Crustacea: Cirripedia), and the larval metamorphosis of a serpulid polycheate, Pomatoleios kraussii Baird, was evaluated. In the case of B. albicostatus, larval mortality increased with an increase in the concentration of Irgarol 1051, and there was a shift in the larval stage targeted from advanced instars to early instars. Nauplii that survived to the cyprid instar stage when reared in the presence of Irgarol 1051 showed prolonged instar and total naupliar duration when compared to the controls. The post-settlement metamorphosis of cyprids significantly varied with Irgarol concentration and also with biofilm age. One and 2-d-old untreated biofilms showed higher metamorphosis when compared to 5-d-old biofilms. However, when the biofilms that promoted cyprid metamorphosis were treated with Irgarol 1051 at low concentrations, metamorphosis rates decreased. Cyprids were prevented from metamorphosing completely by biofilms treated at the highest concentration of Irgarol 1051. Inhibition of metamorphosis was also observed in the case of competent polychaete larvae when exposed to Irgarol 1051 compared to those exposed to metamorphosis inducers such as 3-iso-butyl-1-methylxanthine (IBMX) and natural biofilms. Identification of the pathway(s) that caused the promotory biofilms to become toxic when exposed to Irgarol 1051 is discussed. PMID- 16371338 TI - Quantification of diatoms in biofilms: standardisation of methods. AB - Benthic diatoms, which often dominate marine biofilms are mostly pennate along with a few centric species that have an attached mode of life. Even though the range of diatoms in biofilms is diverse, their ecology is poorly understood because of the difficulty in sampling and enumeration. Scraping or brushing are the traditional methods used for removal of diatoms from biofilms developed on solid substrata. The method of removal is the most critical step in enumerating the biofilm diatom community structure. In this study, a nylon brush and ceramic scraper were used as tools for the removal of diatoms from 1 - 4-day-old biofilms developed on fibreglass coupons and glass microscope slides. Standardisation of methods showed that the sample volume used in the analyses had the least influence on the quantification, whereas the method of removal was critical. The nylon brush was more efficient at recovering diatoms compared to a ceramic scraper. Direct microscopic enumeration of the community in the case of glass slides indicated that scraping resulted in between 30-50% underestimation. Heterogeneity in diatom community structure between replicate samples is one possible reason for such underestimation. PMID- 16371339 TI - Biofilm diatom community structure: influence of temporal and substratum variability. AB - Diatoms, which are early autotrophic colonisers, are an important constituent of the biofouling community in the marine environment. The effects of substratum and temporal variations on the fouling diatom community structure in a monsoon influenced tropical estuary were studied. Fibreglass and glass coupons were exposed every month for a period of 4 days and the diatom population sampled at 24 h intervals, over a period of 14 months. The planktonic diatom community structure differed from the biofilm community. Pennate diatoms dominated the biofilms whilst centric diatoms were dominant in the water column. Among the biofilm diatoms, species belonging to the genera Navicula, Amphora, Nitzschia, Pleurosigma and Thalassionema were dominant. On certain occasions, the influence of planktonic blooms was also seen on the biofilm community. A comparative study of biofilms formed on the two substrata revealed significant differences in density and diversity. However species composition was almost constant. In addition to substratum variations, the biofilm diatom community structure also showed significant seasonal variations, which were attributed to physico-chemical and biological changes in both the water and substratum. Temporal variations in the tychopelagic diatoms of the water were also observed to exert an influence on the biofilm diatom community. Variations in diatom communities may determine the functional ecosystem of the benthic environment. PMID- 16371340 TI - Sublethal effects of a new antifouling candidate on lumpfish (Cyclopterus lumpus L.) and Atlantic cod (Gadus morhua L.) larvae. AB - Sublethal effects of medetomidine, a new generation antifouling compound, on lumpfish (Cyclopterus lumpus L.) and cod (Gadus morhua L.) larvae were examined. The effects on respiration rate and on colour adaptation of newly hatched larvae were assessed after 24-96 h exposure. Exposure of lumpfish larvae to the experimental concentrations resulted in a significant decrease in respiration rate (Lowest Observed Effect Concentration (LOEC) = 5-10 nM) and in the percentage of dark larvae (LOEC = 4 nM). However, no effects on respiration rate of cod larvae were detected. In addition to lumpfish larvae being affected at low concentrations of medetomidine, a reversibility of the effects was observed when 96 h-exposed larvae were incubated in clean seawater for 24-48 h. Considerations relating to the future commercialisation of medetomidine for antifouling purposes are discussed. PMID- 16371341 TI - Survival, growth, settlement and metamorphosis of refrigerated larvae of the mussel Mytilus galloprovincialis Lamarck and their use in settlement and antifouling bioassays. AB - Straight-hinge veliger and pediveliger larvae of the mussel Mytilus galloprovincialis were refrigerated for varying periods for use in bioassays. Straight-hinge veliger larvae grew to the umbo-veliger stage after 2 months in the refrigerator, but no pediveligers were observed during the 3-month refrigeration period. The average survival rate of larvae in the refrigerator was 79% after 1 month, but gradually decreased with the refrigeration period, and was as low as 22% after 3 months. All refrigerated larvae grew to the pediveliger stage in the incubator at 17 degrees C at the same rate as that of the control larvae that were not refrigerated. Settlement and metamorphosis of pediveligers from both refrigerated and control groups were facilitated by microbial film and epinephrine and inhibited by phentolamine. Thus, refrigeration can be used as an effective method of storing larvae of M. galloprovincialis for use in assays to assess candidate settlement inducers and antifouling substances. PMID- 16371342 TI - Integrins in head and neck squamous cell carcinoma (HNSCC): a review of the current literature. AB - Integrins are a family of adhesion molecules performing a major role in multiple cellular functions. Their contribution in carcinogenesis and metastatic process are the object of intense research activity worldwide during the last decades. This review focuses on the existing knowledge about integrin expression in head and neck squamous cell carcinoma (HNSCC) as it has been acquired mainly by immunohistochemical methods and by in vitro assays. The elucidation of the exact role of integrins and the study of expressive alterations of these molecules in cancer cells, may result in novel therapeutical approaches for useful applications in the clinical routine of HNSCC in the future. PMID- 16371343 TI - Roles for Rho/ROCK and vinculin in parietal endoderm migration. AB - The first cell migration event in the mouse embryo is the movement of parietal endoderm cells from the surface of the inner cell mass facing the blastocoel cavity to line the inner surface of the trophectoderm. F9 embryoid bodies provide an in vitro model for this event. They have an inner core of undifferentiated stem cells surrounded by an outer visceral endoderm layer. When plated on a laminin coated substrate, visceral endoderm transitions to parietal endoderm and migrates onto the dish, away from the attached embryoid body. We now show that this outgrowth contains abundant focal complexes and focal adhesions, as well as lamellipodia and filopodia. Treatment with the ROCK inhibitor Y-27632 promotes a 2-fold increase in outgrowth, and a transition from focal adhesions and associated stress fibers, to focal complexes and a decrease in stress fibers. ROCK inhibition also leads to an increase in lamellipodia. Inhibition of RhoA by transfection of a vector encoding C3 transferase, direct administration of the C3 enzyme, or transfection of a vector encoding p190 Rho GTPase Activating Protein also promotes outgrowth and an apparent transition from focal adhesions to focal complexes. Parietal endoderm outgrowth generated using vinculin-deficient F9 stem cells migrates 2-fold further than wild type cultures, but this outgrowth retains the morphology of wild type parietal endoderm, including focal adhesions and stress fibers. Addition of Y-27632 to vinculin-null outgrowth cultures further stimulates migration an additional 2-fold, supporting the conclusion that Rho/ROCK and vinculin regulate parietal endoderm outgrowth by distinct pathways. PMID- 16371344 TI - N-cadherin-mediated cell motility requires cis dimers. AB - Cadherins are expressed on the cell surface as a dimer in the membrane of one cell (cis dimer) that interacts with a cis dimer on an adjacent cell to form an adhesive trans dimer. It is well established that both cis and trans dimers must form for the cadherin to be an effective adhesion protein. In addition to their adhesive activity cadherins also play an important role in modulating cell behavior by regulating cell motility and signal transduction. Whether or not cis or trans dimers are necessary for the nonadhesive functions of cadherins has not been addressed. Here we show that N-cadherin cis dimers are necessary to induce cell motility in epithelial cells and that N-cadherin's ability to modulate the steady state levels of activated small GTPases requires both cis and trans dimers. PMID- 16371345 TI - NOV/CCN3 induces adhesion of muscle skeletal cells and cooperates with FGF2 and IGF-1 to promote proliferation and survival. AB - During mammalian development, expression of the Nephroblastoma overexpressed gene (NOV/CCN3) is tightly regulated in skeletal muscles. Ex vivo, ectopic expression of NOV blocks myogenic differentiation. NOV also supports endothelial cell adhesion and angiogenesis through interactions with integrins. Integrins play fundamental roles during myogenesis. In this study, we show that NOV mediates adhesion and spreading of myoblasts. Myoblasts adhesion to NOV does not require proteoglycans and is dependent on integrin beta1, whereas spreading involves another RGD-sensitive integrin. The C-Terminal part of NOV as well as full-length is able to support adhesion of myoblasts; in addition, both increase focal adhesion kinase (FAK) phosphorylation. Furthermore, NOV is an adhesive substrate that, combined with FGF2 or IGF-1, promotes cell specific proliferation and survival, respectively, in a better way than fibronectin. Taken together, these results identify NOV as an adhesion substrate for myoblasts which, in concert with growth factors, could play a role in the physiology of muscle cells. PMID- 16371346 TI - Inhibition of motile and invasive properties of ovarian cancer cells by ASODN against Rho-associated protein kinase. AB - p160ROCK, a kinase effector of Rho GTPase mediating RhoA-induced assembly of focal adhesions and stress fibers, plays an important role in the invasive process of various tumor cells. The purpose of this study was to investigate the role of p160ROCK in the invasive behaviors of human ovarian cancer cells and their metastasis. Transfection with a dominant-active form of p160ROCK mutant (p160ROCKDelta 3) enhanced cell migration and invasion of ovarian cancer cells, while antisense oligodeoxynucleotide (ASODN) against p160ROCK inhibited the motile and invasive properties of the cells. Our data suggested that p160ROCK was involved in ovarian cancer cell invasion and metastasis by facilitating cancer cell migration, and that p160ROCK might be a potential new effective target for preventing metastasis of ovarian cancer. PMID- 16371347 TI - Structural polymorphism of the HIV-1 leader region explored by computational methods. AB - Experimental studies revealed that the elements of the human immunodeficiency virus type 1 (HIV-1) 5'-untranslated leader region (5'-UTR) can fold in vitro into two alternative conformations, branched (BMH) and 'linearized' (LDI) and switch between them to achieve different functionality. In this study we computationally explored in detail, with our massively parallel genetic algorithm (MPGAfold), the propensity of 13 HIV-1 5'-UTRs to fold into the BMH and the LDI conformation types. Besides the BMH conformations these results predict the existence of two functionally equivalent types of LDI conformations. One is similar to what has been shown in vitro to exist in HIV-1 LAI, the other is a novel conformation exemplified by HIV-1 MAL long-distance interactions. These novel MPGAfold results are further corroborated by a consensus probability matrix algorithm applied to a set of 155 HIV-1 sequences. We also have determined in detail the impact of various strain mutations, domain sizes and folds of elongating sequences simulating folding during transcription on HIV-1 RNA secondary structure folding dynamics. PMID- 16371348 TI - Degradation of endocytosed epidermal growth factor and virally ubiquitinated major histocompatibility complex class I is independent of mammalian ESCRTII. AB - Models for protein sorting at multivesicular bodies in the endocytic pathway of mammalian cells have relied largely on data obtained from yeast. These data suggest the essential role of four ESCRT complexes in multivesicular body protein sorting. However, the putative mammalian ESCRTII complex (hVps25p, hVps22p, and hVps36p) has no proven functional role in endosomal transport. We have characterized the human ESCRTII complex and investigated its function in endosomal trafficking. The human ESCRTII proteins interact with one another, with hVps20p (a component of ESCRTIII), and with their yeast homologues. Our interaction data from yeast two-hybrid studies along with experiments with purified proteins suggest an essential role for the N-terminal domain of hVps22p in the formation of a heterotetrameric ESCRTII complex. Although human ESCRTII is found in the cytoplasm and in the nucleus, it can be recruited to endosomes upon overexpression of dominant-negative hVps4Bp. Interestingly, we find that small interference RNA depletion of mammalian ESCRTII does not affect degradation of epidermal growth factor, a known cargo of the multivesicular body protein sorting pathway. We also show that depletion of the deubiquitinating enzymes AMSH (associated molecule with the SH3 domain of STAM (signal transducing adaptor molecule)) and UBPY (ubiquitin isopeptidase Y) have opposite effects on epidermal growth factor degradation, with UBPY depletion causing dramatic swelling of endosomes. Down-regulation of another cargo, the major histocompatibility complex class I in cells expressing the Kaposi sarcoma-associated herpesvirus protein K3, is unaffected in ESCRTII-depleted cells. Our data suggest that mammalian ESCRTII may be redundant, cargo-specific, or not required for protein sorting at the multivesicular body. PMID- 16371349 TI - Crystal structure of the cytomegalovirus DNA polymerase subunit UL44 in complex with the C terminus from the catalytic subunit. Differences in structure and function relative to unliganded UL44. AB - The human cytomegalovirus DNA polymerase is composed of a catalytic subunit, UL54, and an accessory protein, UL44, which has a structural fold similar to that of other processivity factors, including herpes simplex virus UL42 and homotrimeric sliding clamps such as proliferating cell nuclear antigen. Several specific residues in the C-terminal region of UL54 and in the "connector loop" of UL44 are required for the association of these proteins. Here, we describe the crystal structure of residues 1-290 of UL44 in complex with a peptide from the extreme C terminus of UL54, which explains this interaction at a molecular level. The UL54 peptide binds to structural elements similar to those used by UL42 and the sliding clamps to associate with their respective binding partners. However, the details of the interaction differ from those of other processivity factor peptide complexes. Crucial residues include a three-residue hydrophobic "plug" from the UL54 peptide and Ile(135) of UL44, which forms a critical intramolecular hydrophobic anchor for interactions between the connector loop and the peptide. As was the case for the unliganded UL44 structure, the UL44-peptide complex forms a head-to-head dimer that could potentially form a C-shaped clamp on DNA. However, the peptide-bound structure displays subtle differences in the relative orientation of the two subdomains of the protein, resulting in a more open clamp, which we predicted would affect its association with DNA. Indeed, filter binding assays revealed that peptide-bound UL44 binds DNA with higher affinity. Thus, interaction with the catalytic subunit appears to affect both the structure and function of UL44. PMID- 16371350 TI - Targeting and trafficking of the human thiamine transporter-2 in epithelial cells. AB - Humans lack biochemical pathways for thiamine synthesis, so cellular requirements are met via specific carrier-mediated uptake pathways. Two proteins from the solute carrier SLC19A gene family have been identified as human thiamine transporters (hTHTRs), SLC19A1 (hTHTR1) and SLC19A2 (hTHTR2). Both of these transporters are co-expressed but are differentially targeted in polarized cell types that mediate vectorial thiamine transport (e.g. renal and intestinal epithelia). It is important to understand the domain structure of these proteins, namely which regions within the polypeptide sequence are important for physiological delivery to the cell surface, in order to understand the impact of clinically relevant mutations on thiamine transport. Here we have characterized the mechanisms regulating hTHTR2 distribution by using live cell imaging methods that resolve the targeting and trafficking dynamics of full-length hTHTR2, a series of hTHTR2 truncation mutants, as well as chimeras comprising the hTHTR1 and hTHTR2 sequence. We showed the following: (i) that the cytoplasmic COOH-tail of hTHTR2 is not essential for apical targeting in polarized cells; (ii) that delivery of hTHTR2 to the cell surface is critically dependent on the integrity of the transmembrane backbone of the polypeptide so that minimal truncations abrogate cell surface expression of hTHTR2; and (iii) video rate images of hTHTR2 containing intracellular vesicles displayed rapid bi-directional trafficking events to and from the cell surface impaired by microtubule-disrupting but not microfilament-disrupting agents as well as by overexpression of the dynactin subunit dynamitin (p50). Finally, we compared the behavior of hTHTR2 with that of hTHTR1 and the human reduced folate carrier (SLC19A1) to underscore commonalities in the cell surface targeting mechanisms of the entire SLC19A gene family. PMID- 16371351 TI - A downshift in temperature activates the high osmolarity glycerol (HOG) pathway, which determines freeze tolerance in Saccharomyces cerevisiae. AB - The molecular mechanisms that enable yeast cells to detect and transmit cold signals and their physiological significance in the adaptive response to low temperatures are unknown. Here, we have demonstrated that the MAPK Hog1p is specifically activated in response to cold. Phosphorylation of Hog1p was dependent on Pbs2p, the MAPK kinase (MAPKK) of the high osmolarity glycerol (HOG) pathway, and Ssk1p, the response regulator of the two-component system Sln1p Ypd1p. However, Sho1p was not required. Interestingly, phosphorylation of Hog1p was stimulated at 30 degrees C in cells exposed to the membrane rigidifier agent dimethyl sulfoxide. Moreover, Hog1p activation occurred specifically through the Sln1 branch. This suggests that Sln1p monitors changes in membrane fluidity caused by cold. Quite remarkably, activation of Hog1p at low temperatures affected the transcriptional response to cold shock. Indeed, the absence of Hog1p impaired the cold-instigated expression of genes for trehalose- and glycerol synthesizing enzymes and small chaperones. Moreover, a downward transfer to 12 or 4 degrees C stimulated the overproduction of glycerol in a Hog1p-dependent manner. However, hog1Delta mutant cells showed no growth defects at 12 degrees C as compared with the wild type. On the contrary, deletion of HOG1 or GPD1 decreased tolerance to freezing of wild-type cells preincubated at a low temperature, whereas no differences could be detected in cells shifted directly from 30 to -20 degrees C. Thus, exposure to low temperatures triggered a Hog1p dependent accumulation of glycerol, which is essential for freeze protection. PMID- 16371352 TI - Expression of cyclooxygenase-2 is regulated by glycogen synthase kinase-3beta in gastric cancer cells. AB - Cyclooxygenase-2 (COX-2) expression is a marker of poor prognosis in gastric cancer patients, and its inhibition suppresses gastric tumorigenesis in experimental animal models. The mechanism that leads to COX-2 overexpression in this tumor type is unknown. We have now shown that inhibition of phosphatidylinositol 3-kinase by LY294002 suppresses both basal and phorbol myristate acetate-induced COX-2 expression in TMK-1 and MKN-28 gastric cancer cells. Furthermore, inhibition of glycogen synthase kinase-3beta (GSK-3beta) by SB415286 induced expression of COX-2 mRNA and protein as well as the enzyme activity in the gastric cancer cells. The effect of SB415286 was confirmed by the use of two additional GSK-3beta inhibitors, lithium chloride and SB216763. SB415286 had a modest 1.6-fold stimulatory effect on a 2-kb COX-2 promoter reporter construct, but more importantly, it was shown to block the decay of COX 2 mRNA. In contrast to modulation of phosphatidylinositol 3-kinase/Akt/GSK-3beta pathway, inhibitors of mitogen-activated protein kinases (MEK 1/2, p38, JNK) or the mammalian target of rapamycin did not alter COX-2 expression in gastric cancer cells. Our data show that inhibition of GSK-3beta stimulates COX-2 expression in gastric cancer cells, which seems to be primarily facilitated via an increase in mRNA stability and to a lesser extent through enhanced transcription. PMID- 16371353 TI - A rostrocaudal muscular dystrophy caused by a defect in choline kinase beta, the first enzyme in phosphatidylcholine biosynthesis. AB - Muscular dystrophies include a diverse group of genetically heterogeneous disorders that together affect 1 in 2000 births worldwide. The diseases are characterized by progressive muscle weakness and wasting that lead to severe disability and often premature death. Rostrocaudal muscular dystrophy (rmd) is a new recessive mouse mutation that causes a rapidly progressive muscular dystrophy and a neonatal forelimb bone deformity. The rmd mutation is a 1.6-kb intragenic deletion within the choline kinase beta (Chkb) gene, resulting in a complete loss of CHKB protein and enzymatic activity. CHKB is one of two mammalian choline kinase (CHK) enzymes (alpha and beta) that catalyze the phosphorylation of choline to phosphocholine in the biosynthesis of the major membrane phospholipid phosphatidylcholine. While mutant rmd mice show a dramatic decrease of CHK activity in all tissues, the dystrophy is only evident in skeletal muscle tissues in an unusual rostral-to-caudal gradient. Minor membrane disruption similar to dysferlinopathies suggest that membrane fusion defects may underlie this dystrophy, because severe membrane disruptions are not evident as determined by creatine kinase levels, Evans Blue infiltration, and unaltered levels of proteins in the dystrophin-glycoprotein complex. The rmd mutant mouse offers the first demonstration of a defect in a phospholipid biosynthetic enzyme causing muscular dystrophy, representing a unique model for understanding mechanisms of muscle degeneration. PMID- 16371354 TI - Tuftsin binds neuropilin-1 through a sequence similar to that encoded by exon 8 of vascular endothelial growth factor. AB - Tuftsin, Thr-Lys-Pro-Arg (TKPR), is an immunostimulatory peptide with reported nervous system effects as well. We unexpectedly found that tuftsin and a higher affinity antagonist, TKPPR, bind selectively to neuropilin-1 and block vascular endothelial growth factor (VEGF) binding to that receptor. Dimeric and tetrameric forms of TKPPR had greatly increased affinity for neuropilin-1 based on competition binding experiments. On endothelial cells tetrameric TKPPR inhibited the VEGF(165)-induced autophosphorylation of vascular endothelial growth factor receptor-2 (VEGFR-2) even though it did not directly inhibit VEGF binding to VEGFR-2. Homology between exon 8 of VEGF and TKPPR suggests that the sequence coded for by exon 8 may stabilize VEGF binding to neuropilin-1 to facilitate signaling through VEGFR-2. Given the overlap between processes involving neuropilin-1 and tuftsin, we propose that at least some of the previously reported effects of tuftsin are mediated through neuropilin-1. PMID- 16371355 TI - The second member of the human and murine bubblegum family is a testis- and brainstem-specific acyl-CoA synthetase. AB - Acyl-CoA synthetases that activate fatty acids to their CoA derivatives play a central role in fatty acid metabolism. ACSBG1, an acyl-CoA synthetase originally identified in the fruit fly mutant bubblegum, was hypothesized to contribute to the biochemical pathology of X-linked adrenoleukodystrophy. We looked for homologous proteins and identified ACSBG2 in humans, mice, and rats. Human ACSBG1 and ACSBG2 amino acid sequences are 50% identical. ACSBG2 expression was confined to the testis and brainstem. Immunohistochemistry and in situ hybridization studies further localized ACSBG2 expression to testicular Sertoli cells and large motoneurons in the medulla oblongata and cervical spinal cord. Full-length cDNA encoding human and mouse ACSBG2 was cloned. In transfected COS-1 cells, both human and murine ACSBG2 were detected as 75- to 80-kDa proteins by Western blot. Cells overexpressing ACSBG2 had increased ability to activate oleic acid (C18:1omega9) and linoleic acid (C18:2omega6) but not other fatty acid substrates tested. Within a highly conserved motif known to be important for catalysis, human ACSBG2 contains a histidine residue where all other known acyl-CoA synthetases, including mouse and rat ACSBG2, contain an arginine. This substitution resulted in a shift of the human ACSBG2 pH optimum to a more acidic pH. Mutation of this histidine to arginine improved catalytic function at neutral pH by shifting the pH profile without affecting substrate specificity. Although the role of ACSBG2 in testicular and neuronal lipid metabolism remains unclear, the limited tissue expression pattern and limited substrate specificity rule out a likely role for this enzyme in X-linked adrenoleukodystrophy pathology. PMID- 16371356 TI - Ligands for the beta-glucan receptor, Dectin-1, assigned using "designer" microarrays of oligosaccharide probes (neoglycolipids) generated from glucan polysaccharides. AB - Dectin-1 is a C-type lectin-like receptor on leukocytes that mediates phagocytosis and inflammatory mediator production in innate immunity to fungal pathogens. Dectin-1 lacks residues involved in calcium ligation that mediates carbohydrate-binding by classical C-type lectins; nevertheless, it binds zymosan, a particulate beta-glucan-rich extract of Saccharomyces cerevisiae, and binding is inhibited by polysaccharides rich in beta1,3- or both beta1,3- and beta1,6 linked glucose. The oligosaccharide ligands on glucans recognized by Dectin-1 have not yet been delineated precisely. It is also not known whether Dectin-1 can interact with other types of carbohydrates. We have investigated this, since Dectin-1 shows glucan-independent binding to a subset of T-lymphocytes and is involved in triggering their proliferation. Here we assign oligosaccharide ligands for Dectin-1 using the neoglycolipid-based oligosaccharide microarray technology, a unique approach for constructing microarrays of lipid-linked oligosaccharide probes from desired sources. We generate "designer" microarrays from three glucan polysaccharides, a neutral soluble glucan isolated from S. cerevisiae and two bacterial glucans, curdlan from Alcaligenes faecalis and pustulan from Umbilicaria papullosa, and use these in conjunction with 187 diverse, sequence-defined, predominantly mammalian-type, oligosaccharide probes. Among these, Dectin-1 binding is detected exclusively to 1,3-linked glucose oligomers, the minimum length required for detectable binding being a 10- or 11 mer. Thus, the ligands assigned so far are exogenous rather than endogenous. We further show that Dectin-1 ligands, 11-13 gluco-oligomers, in clustered form (displayed on liposomes), mimic the macromolecular beta-glucans and compete with zymosan binding and triggering of tumor necrosis factor-alpha secretion by a Dectin-1-expressing macrophage cell line. PMID- 16371357 TI - Inheritable histone H4 acetylation of somatic chromatins in cloned embryos. AB - A viable cloned animal indicates that epigenetic status of the differentiated cell nucleus is reprogrammed to an embryonic totipotent state. However, molecular events regarding epigenetic reprogramming of the somatic chromatin are poorly understood. Here we provide new insight that somatic chromatins are refractory to reprogramming of histone acetylation during early development. A low level of acetylated histone H4-lysine 5 (AcH4K5) of the somatic chromatin was sustained at the pronuclear stage. Unlike in vitro fertilized (IVF) embryos, the AcH4K5 level remarkably reduced at the 8-cell stage in cloned bovine embryos. The AcH4K5 status of somatic chromatins transmitted to cloned and even recloned embryos. Differences of AcH4K5 signal intensity were more distinguishable in the metaphase chromosomes between IVF and cloned embryos. Two imprinted genes, Ndn and Xist, were aberrantly expressed in cloned embryos as compared with IVF embryos, which is partly associated with the AcH4K5 signal intensity. Our findings suggest that abnormal epigenetic reprogramming in cloned embryos may be because of a memory mechanism, the epigenetic status itself of somatic chromatins. PMID- 16371358 TI - 3-nitropropionic acid is a suicide inhibitor of mitochondrial respiration that, upon oxidation by complex II, forms a covalent adduct with a catalytic base arginine in the active site of the enzyme. AB - We report three new structures of mitochondrial respiratory Complex II (succinate ubiquinone oxidoreductase, E.C. 1.3.5.1) at up to 2.1 A resolution, with various inhibitors. The structures define the conformation of the bound inhibitors and suggest the residues involved in substrate binding and catalysis at the dicarboxylate site. In particular they support the role of Arg(297) as a general base catalyst accepting a proton in the dehydrogenation of succinate. The dicarboxylate ligand in oxaloacetate-containing crystals appears to be the same as that reported for Shewanella flavocytochrome c treated with fumarate. The plant and fungal toxin 3-nitropropionic acid, an irreversible inactivator of succinate dehydrogenase, forms a covalent adduct with the side chain of Arg(297). The modification eliminates a trypsin cleavage site in the flavoprotein, and tandem mass spectroscopic analysis of the new fragment shows the mass of Arg(297) to be increased by 83 Da and to have the potential of losing 44 Da, consistent with decarboxylation, during fragmentation. PMID- 16371359 TI - Solution structure and DNA-binding mode of the matrix attachment region-binding domain of the transcription factor SATB1 that regulates the T-cell maturation. AB - SATB1 is a transcriptional regulator controlling the gene expression that is essential in the maturation of the immune T-cell. SATB1 binds to the nuclear matrix attachment regions of DNA, where it recruits histone deacetylase and represses transcription through a local chromatin remodeling. Here we determined the solution structure of the matrix attachment region-binding domain, possessing similarity to the CUT DNA-binding domain, of human SATB1 by NMR spectroscopy. The structure consists of five alpha-helices, in which the N-terminal four are arranged similarly to the four-helix structure of the CUT domain of hepatocyte nuclear factor 6alpha. By an NMR chemical shift perturbation analysis and by surface plasmon resonance analyses of SATB1 mutant proteins, an interface for DNA binding was revealed to be located at the third helix and the surrounding regions. Surface plasmon resonance experiments using groove-specific binding drugs and methylated DNAs indicated that the domain recognizes DNA from the major groove side. These observations suggested that SATB1 possesses a DNA-binding mode similar to that of the POU-specific DNA-binding domain, which is known to share structural similarity to the four-helix CUT domain. PMID- 16371360 TI - The role of ribosomal protein L11 in class I release factor-mediated translation termination and translational accuracy. AB - It has been suggested from in vivo and cryoelectron micrographic studies that the large ribosomal subunit protein L11 and its N-terminal domain play an important role in peptide release by, in particular, the class I release factor RF1. In this work, we have studied in vitro the role of L11 in translation termination with ribosomes from a wild type strain (WT-L11), an L11 knocked-out strain (DeltaL11), and an L11 N terminus truncated strain (Cter-L11). Our data show 4-6 fold reductions in termination efficiency (k(cat)/K(m)) of RF1, but not of RF2, on DeltaL11 and Cter-L11 ribosomes compared with wild type. There is, at the same time, no effect of these L11 alterations on the maximal rate of ester bond cleavage by either RF1 or RF2. The rates of dissociation of RF2 but not of RF1 from the ribosome after peptide release are somewhat reduced by the L11 changes irrespective of the presence of RF3, and they cause a 2-fold decrease in the missense error. Our results suggest that the L11 modifications increase nonsense suppression at UAG codons because of the reduced termination efficiency of RF1 and that they decrease nonsense suppression at UGA codons because of a decreased missense error level. PMID- 16371361 TI - 4'-phosphopantetheine biosynthesis in Archaea. AB - Coenzyme A as the principal acyl carrier is required for many synthetic and degradative reactions in intermediary metabolism. It is synthesized in five steps from pantothenate, and recently the CoaA biosynthetic genes of eubacteria, plants, and human were all identified and cloned. In most bacteria, the so-called Dfp proteins catalyze the synthesis of the coenzyme A precursor 4' phosphopantetheine. Dfp proteins are bifunctional enzymes catalyzing the synthesis of 4'-phosphopantothenoylcysteine (CoaB activity) and its decarboxylation to 4'-phosphopantetheine (CoaC activity). Here, we demonstrate the functional characterization of the CoaB and CoaC domains of an archaebacterial Dfp protein. Both domains of the Methanocaldococcus jannaschii Dfp protein were purified as His tag proteins, and their enzymatic activities were then identified and characterized by site-directed mutagenesis. Although the nucleotide binding motif II of the CoaB domain resembles that of eukaryotic enzymes, Methanocaldococcus CoaB is a CTP- and not an ATP-dependent enzyme, as shown by detection of the 4'-phosphopantothenoyl-CMP intermediate. The proposed 4'-phosphopantothenoylcysteine binding clamp of the Methanocaldococcus CoaC activity differs significantly from those of other characterized CoaC proteins. In particular, the active site cysteine residue, which otherwise is involved in the reduction of an aminoenethiol reaction intermediate, is not present. Moreover, the conserved Asn residue of the PXMNXXMW motif, which contacts the carboxyl group of 4'-phosphopantothenoylcysteine, is exchanged for His. PMID- 16371362 TI - Huntingtin and mutant SOD1 form aggregate structures with distinct molecular properties in human cells. AB - Expression of many proteins associated with neurodegenerative disease results in the appearance of misfolded species that readily adopt alternate folded states. In vivo, these appear as punctated subcellular structures typically referred to as aggregates or inclusion bodies. Whereas groupings of these distinct proteins into a common morphological class have been useful conceptually, there is some suggestion that aggregates are not homogeneous and can exhibit a range of biological properties. In this study, we use dynamic imaging analysis of living cells to compare the aggregation and growth properties of mutant huntingtin with polyglutamine expansions or mutant SOD1 (G85R/G93A) to examine the formation of aggregate structures and interactions with other cellular proteins. Using a dual conditional expression system for sequential expression of fluorescence-tagged proteins, we show that mutant huntingtin forms multiple intracellular cytoplasmic and nuclear structures composed of a dense core inaccessible to nascent polypeptides surrounded by a surface that stably sequesters certain transcription factors and interacts transiently with molecular chaperones. In contrast, mutant SOD1 (G85R/G93A) forms a distinct aggregate structure that is porous, through which nascent proteins diffuse. These results reveal that protein aggregates do not correspond to a single common class of subcellular structures, and rather that there may be a wide range of aggregate structures, perhaps each corresponding to the specific disease-associated protein with distinct consequences on the biochemical state of the cell. PMID- 16371363 TI - Human alveolar macrophages are deficient in PTEN. The role of endogenous oxidants. AB - Human alveolar macrophages play a critical role in host defense and in the development of inflammation and fibrosis in the lung. Unlike their precursor cells, blood monocytes, alveolar macrophages are long-lived and tend to be resistant to apoptotic stimuli. In this study, we examined the role of differentiation in altering baseline phosphatidylinositol (PI) 3-kinase/Akt activity. We found that differentiation increased activity of pro-survival PI 3 kinase/Akt while decreasing amounts of the negative PI 3-kinase regulator, PTEN. PTEN is a lipid phosphatase with activity against phosphatidylinositol 3,4,5 trisphosphate (PI(3,4,5)P3), the major bioactive product of PI 3-kinase. Examining in vivo differentiation of alveolar macrophages (by comparing blood monocytes to alveolar macrophages from single donors), we found that differentiation resulted in increased baseline reactive oxygen species (ROS) in the alveolar macrophages. This led to a deficiency in PTEN, increased activity of Akt, and prolonged survival of alveolar macrophages. These data support the hypothesis that alterations in ROS levels contribute to macrophage homeostasis by altering the balance between PI 3-kinase/Akt and the phosphatase, PTEN. PMID- 16371364 TI - The 2',5'-oligoadenylate synthetase 1b is a potent inhibitor of West Nile virus replication inside infected cells. AB - The 2',5'-oligoadenylate synthetase (OAS) proteins associated with endoribonuclease RNase L are components of the interferon-regulated OAS/RNase L system, which is an RNA decay pathway known to play an important role in the innate antiviral immunity. A large body of evidence suggests a critical role for the 1b isoform of the mouse Oas gene (Oas1b) in resistance to West Nile virus (WNV) infection in vivo. WNV is a positive, single-stranded RNA virus responsible for severe encephalitis in a large range of animal species and humans. To investigate the molecular basis for the sensitivity of WNV to the Oas1b antiviral pathway, we established a stable mouse fibroblastic cell clone that up-regulates Oas1b protein expression under the control of the Tet-Off expression system. We showed that murine cells respond to Oas1b expression by efficiently inhibiting WNV replication. The antiviral action of Oas1b was essentially restricted to the early stages in virus life cycle. We found that the inability of WNV to productively infect the Oas1b-expressing cells was attributable to a dramatic reduction in positive-stranded viral RNA level. Thus, Oas1b represents an antiviral pathway that exerts its inhibitory effect on WNV replication by preventing viral RNA accumulation inside infected cells. PMID- 16371365 TI - Amino acid changes in Drosophila alphaPS2betaPS integrins that affect ligand affinity. AB - We developed a ligand-mimetic antibody Fab fragment specific for Drosophila alphaPS2betaPS integrins to probe the ligand binding affinities of these invertebrate receptors. TWOW-1 was constructed by inserting a fragment of the extracellular matrix protein Tiggrin into the H-CDR3 of the alphavbeta3 ligand mimetic antibody WOW-1. The specificity of alphaPS2betaPS binding to TWOW-1 was demonstrated by numerous tests used for other integrin-ligand interactions. Binding was decreased in the presence of EDTA or RGD peptides and by mutation of the TWOW-1 RGD sequence or the betaPS metal ion-dependent adhesion site (MIDAS) motif. TWOW-1 binding was increased by mutations in the alphaPS2 membrane proximal cytoplasmic GFFNR sequence or by exposure to Mn2+. Although Mn2+ is sometimes assumed to promote maximal integrin activity, TWOW-1 binding in Mn2+ could be increased further by the alphaPS2 GFFNR --> GFANA mutation. A mutation in the betaPS I domain (betaPS-b58; V409D) greatly increased ligand binding affinity, explaining the increased cell spreading mediated by alphaPS2betaPS-b58. Further mutagenesis of this residue suggested that Val-409 normally stabilizes the closed head conformation. Mutations that potentially reduce interaction of the integrin beta subunit plexin-semaphorin-integrin (PSI) and stalk domains have been shown to have activating properties. We found that complete deletion of the betaPS PSI domain enhanced TWOW-1 binding. Moreover the PSI domain is dispensable for at least some other integrin functions because betaPS-DeltaPSI displayed an enhanced ability to mediate cell spreading. These studies establish a means to evaluate mechanisms and consequences of integrin affinity modulation in a tractable model genetic system. PMID- 16371366 TI - The tumor suppressor PTEN is necessary for human Sprouty 2-mediated inhibition of cell proliferation. AB - Sprouty family proteins are novel regulators of growth factor actions. Human Sprouty 2 (hSPRY2) inhibits the proliferation of a number of different cell types. However, the mechanisms involved in the anti-proliferative actions of hSPRY2 remain to be elucidated. Here we have demonstrated that hSPRY2 increases the amount of the tumor suppressor phosphatase and tensin homolog deleted on chromosome 10 (PTEN) and decreases its phosphorylation. The resultant increase in PTEN activity is reflected in decreased activation of Akt by epidermal growth factor and serum. Consistent with increased PTEN activity, in hSPRY2-expressing cells, the progression of cells from the G1 to S phase is decreased. By using PTEN null primary mouse embryonic fibroblasts and their isogenic controls as well as small interfering RNA against PTEN, we demonstrated that PTEN is necessary for hSPRY2 to inhibit Akt activation by epidermal growth factor as well as cell proliferation. Overall, we concluded that hSPRY2 mediates its anti-proliferative actions by altering PTEN content and activity. PMID- 16371368 TI - Effects of a leukemia-associated gain-of-function mutation of SHP-2 phosphatase on interleukin-3 signaling. AB - Mutations in SHP-2 phosphatase that cause hyperactivation of its catalytic activity have been identified in human leukemias, particularly juvenile myelomonocytic leukemia, which is characterized by hypersensitivity of myeloid progenitor cells to granulocyte macrophage colony-stimulating factor and interleukin (IL)-3. However, the molecular mechanisms by which gain-of-function (GOF) mutations of SHP-2 induce hematopoietic malignancies are not fully understood. Our previous studies have shown that SHP-2 plays an essential role in IL-3 signal transduction in both catalytic-dependent and -independent manners and that overexpression (5-6-fold) of wild type (WT) SHP-2 attenuates IL-3-mediated hematopoietic cell function through accelerated dephosphorylation of STAT5. These results raised the possibility that SHP-2-associated leukemias are not solely attributed to the increased catalytic activity of GOF mutant SHP-2. GOF mutant SHP-2 must have gained additional capacities. To test this possibility, we investigated effects of a GOF mutation of SHP-2 (SHP-2 E76K) on hematopoietic cell function and IL-3 signal transduction by comparing with those of overexpressed WT SHP-2. Our results showed that SHP-2 E76K mutation caused myeloproliferative disease in mice, while overexpression of WT SHP-2 decreased hematopoietic potential of the transduced cells in recipient animals. The E76K mutation in the N-terminal Src homology 2 domain increased interactions of mutant SHP-2 with Grb2, Gab2, and p85, leading to hyperactivation of IL-3-induced Erk and phosphatidylinositol 3-kinase (PI3K) pathways. In addition, despite the substantial increase in the catalytic activity, dephosphorylation of STAT5 by SHP 2 E76K was dampened. Furthermore, catalytically inactive SHP-2 E76K with an additional C459S mutation retained the capability to increase the interaction with Gab2 and to enhance the activation of the PI3K pathway. Taken together, these studies suggest that in addition to the elevated catalytic activity, fundamental changes in physical and functional interactions between GOF mutant SHP-2 and signaling partners also play an important role in SHP-2-related leukemigenesis. PMID- 16371367 TI - Regulation of nuclear translocation of HDAC3 by IkappaBalpha is required for tumor necrosis factor inhibition of peroxisome proliferator-activated receptor gamma function. AB - Inhibition of peroxisome proliferator-activated receptor gamma (PPARgamma) function by TNF-alpha contributes to glucose and fatty acid metabolic disorders in inflammation and cancer, although the molecular mechanism is not fully understood. In this study, we demonstrate that nuclear translocation of HDAC3 is regulated by TNF-alpha, and this event is required for inhibition of transcriptional activity of PPARgamma by TNF-alpha. HDAC3 is associated with IkappaBalpha in the cytoplasm. After IkappaBalpha degradation in response to TNF alpha, HDAC3 is subject to nuclear translocation, leading to an increase in HDAC3 activity in the nucleus. This event leads to subcellular redistribution of HDAC3. Knock-out of IkappaBalpha, but not p65 or p50, leads to disappearance of HDAC3 in the cytoplasm, which is associated with HDAC3 enrichment in the nucleus. These data suggest that inhibition of PPARgamma by TNF-alpha is not associated with a reduction in the DNA binding activity of PPARgamma. Rather, these results suggest that IkappaBalpha-dependent nuclear translocation of HDAC3 is responsible for PPARgamma inhibition by TNF-alpha. PMID- 16371369 TI - Release of free F2-isoprostanes from esterified phospholipids is catalyzed by intracellular and plasma platelet-activating factor acetylhydrolases. AB - F2-isoprostanes are produced in vivo by nonenzymatic peroxidation of arachidonic acid esterified in phospholipids. Increased urinary and plasma F2-isoprostane levels are associated with a number of human diseases. These metabolites are regarded as excellent markers of oxidant stress in vivo. Isoprostanes are initially generated in situ, i.e. when the arachidonate precursor is esterified in phospholipids, and they are subsequently released in free form. Although the mechanism(s) responsible for the release of free isoprostanes after in situ generation in membrane phospholipids is, for the most part, unknown, this process is likely mediated by phospholipase A2 activity(ies). Here we reported that human plasma contains an enzymatic activity that catalyzes this reaction. The activity associates with high density and low density lipoprotein and comigrates with platelet-activating factor (PAF) acetylhydrolase on KBr density gradients. Plasma samples from subjects deficient in PAF acetylhydrolase do not release F2 isoprostanes from esterified precursors. The intracellular PAF acetylhydrolase II, which shares homology to the plasma enzyme, also catalyzes this reaction. We found that both the intracellular and plasma PAF acetylhydrolases have high affinity for esterified F2-isoprostanes. However, the rate of esterified F2 isoprostane hydrolysis is much slower compared with the rate of hydrolysis of other substrates utilized by these enzymes. Studies using PAF acetylhydrolase transgenic mice indicated that these animals have a higher capacity to release F2 isoprostanes compared with nontransgenic littermates. Our results suggested that PAF acetylhydrolases play key roles in the hydrolysis of F2-isoprostanes esterified on phospholipids in vivo. PMID- 16371371 TI - Two new theories of autism: hyper-systemising and assortative mating. PMID- 16371372 TI - Influenza related hospital admissions in children: evidence about the burden keeps growing but the route to policy change remains uncertain. PMID- 16371373 TI - Influenza related hospitalisations in Sydney, New South Wales, Australia. AB - BACKGROUND: Routine influenza vaccination for children aged 6-23 months has recently been recommended in the United States. Accurate assessment of influenza related burden of illness in children could support similar recommendations in other settings. However, routinely available data underestimate the role of influenza in causing hospitalisation, and indirect estimation methods face difficulties controlling for the concurrent circulation of respiratory syncytial virus (RSV). Recent studies from Hong Kong and the United States have used differing methods to estimate the true burden of influenza related hospitalisation, with disparate results. METHODS: Retrospective population based study of children less than 18 years of age from Sydney, Australia, 1994 to 2001. Using two previously reported methods, estimates of annual hospitalisation rates attributable to influenza were derived by comparison of mean hospitalisation rates for acute respiratory disease during periods of high influenza activity and low RSV activity (defined using virological surveillance data) and periods where both influenza and RSV activity were low. These estimates were compared to rates of hospitalisation where influenza was recorded as the principal discharge diagnosis. RESULTS: Hospitalisation rates attributable to influenza were up to 11 times higher, depending on the age group and method used, compared to rates calculated from principal discharge diagnosis codes. CONCLUSIONS: Although there remains considerable uncertainty in estimating influenza related morbidity by methods using excess hospitalisations, even minimum estimates of disease burden warrant consideration of routine influenza immunisation for all children less than 2 years of age. Such estimates, derived from principal discharge diagnosis codes, are available in most settings. PMID- 16371374 TI - Chronic recurrent multifocal osteomyelitis is a differential of childhood limp. PMID- 16371375 TI - Smuggling contraband drugs using paediatric "body packers". PMID- 16371376 TI - Pre-pubertal growth and cognitive function. AB - British longitudinal data were used to investigate the association of heights at 22 months and 5 years with a digit recall test at age 10 years. Greater height, particularly at 5 years, was associated with higher scores, suggesting that some exposures influence both growth and capability for cognitive function. PMID- 16371377 TI - Clinical and molecular findings in IPEX syndrome. AB - IPEX (immunodysregulation, polyendocrinopathy, enteropathy, X linked syndrome) is a rare disorder which usually results in death in early infancy or childhood. Clinical awareness remains the cornerstone of diagnosis, and provided that the diagnosis is entertained, mutation analysis for FOXP3 gene mutations can be confirmatory. Two new patients in whom IPEX was diagnosed retrospectively are reported. PMID- 16371378 TI - How to make the RCPCH CPD guidelines work for you. AB - Continuing professional development (CPD) is defined as a systematic process of lifelong learning and professional development. Its aim is to enable career grade doctors (consultants, associate specialists, staff grades, and their equivalents) to maintain and enhance their knowledge, skills, and competence for effective clinical practice to meet the needs of children. This article is designed to assist and improve acquisition of CPD. PMID- 16371379 TI - What's happening in postgraduate medical education? AB - "It is a secret, both in nature and state, that it is safer to change many things at once." (Francis Bacon, "Of Regiment of Health", Essays, 1625). PMID- 16371380 TI - The Royal College of Paediatrics and Child Health programme for subspecialty training. AB - The National Grid is a training scheme that represents a unique partnership between Deaneries and the Royal College of Paediatrics and Child Health (RCPCH) in offering through national competition, equitable access to high quality subspecialty training. Paediatrics is unusual in that within the umbrella Certificate of Completion of Specialist Training in paediatrics there is the potential to train in one of 13 recognised paediatric subspecialties. PMID- 16371381 TI - Towards evidence based medicine for paediatricians. PMID- 16371382 TI - Should a neonate with possible late onset infection always have a lumbar puncture? PMID- 16371383 TI - Does continuous insulin infusion improve glycaemic control and nutrition in hyperglycaemic very low birth weight infants? PMID- 16371384 TI - Should premedication be used for semi-urgent or elective intubation in neonates? PMID- 16371385 TI - To all those who know nothing about diabetes. PMID- 16371386 TI - Physician, philosopher, and paediatrician: John Locke's practice of child health care. AB - G.F. Still's History of Paediatrics restricted the philosopher John Locke's (1632 1704) influence in paediatrics to pedagology and specifically his Some Thoughts Concerning Education (1693). This significantly limits Locke's immense ongoing influence on child health care and human rights. Locke was a physician and had a lifelong interest in medicine. His case records and journals relate some of his paediatric cases. His correspondence includes letters from Thomas Sydenham, the "English Hippocrates" (1624-89) when Locke has sought advice on a paediatric case as well as other correspondence from parents regarding child health care and management of learning disability. Locke assisted and influenced Thomas Sydenham with his writing, and Locke's own work, Two Treatises on Government, clearly stated the rights of children and limitation of parental authority. Furthermore, Locke's thoughts on Poor Law, making an economic case for a workhouse in every parish, were implemented from 1834. PMID- 16371387 TI - Edgar Schoen does not represent the North American view of male circumcision. PMID- 16371388 TI - Gaviscon for gastro-oesophageal reflux in infants: a poorly effective treatment? PMID- 16371389 TI - Neonate with staphylococcal scalded skin syndrome. PMID- 16371390 TI - Blood flow in the common carotid artery in term and preterm infants: reproducibility and relation to cardiac output. AB - AIM: To assess the reproducibility of, and determine normative data for, flow volume measurements from the right common carotid artery (CCA) and its relation to left ventricular output (LVO) in stable term and preterm babies using Doppler ultrasound. METHODS: Right CCA flow volume was measured using a near focus, high frequency transducer by obtaining intensity weighted mean velocity and right CCA diameter. LVO was determined using standard Doppler techniques. Reproducibility studies were performed on 30 newborn infants by two observers. Normative data were obtained from 40 spontaneously breathing preterm babies and 21 term babies. RESULTS: The intraobserver coefficient of variation for CCA flow measurements was 10.5% for observer 1 and 15.4% for observer 2, whereas the interobserver coefficient of variation was 16.4%. In term and preterm infants, right CCA flow was about 20 ml/kg/min, accounting for 11% of cardiac output. Among the preterm infants, there was a positive correlation of right CCA flow with gestation (r = 0.61, p<0.001), weight (r = 0.64, p<0.001), and LVO (r = 0.59, p<0.001). Right CCA diameter also increased with weight (r = 0.63, p<0.001) and gestation (r = 0.58, p<0.001). The proportion of LVO distributed to the right CCA did not increase with gestation, nor did the right CCA flow per kg body weight. CONCLUSIONS: It is possible to perform reproducible measurements of flow volume in the CCA of newborn infants. In stable, spontaneously breathing babies, both cardiac output and carotid flow increased with gestation and body weight. The proportion of cardiac output distributed to the right CCA remained relatively constant across gestation. PMID- 16371391 TI - Umbilical cord blood procalcitonin and C reactive protein concentrations as markers for early diagnosis of very early onset neonatal infection. AB - Procalcitonin (PCT) and C reactive protein (CRP) concentrations in umbilical cord blood of 197 neonates were measured to evaluate their value as markers of infection. Sixteen of the neonates were infected. The sensitivity, specificity, and negative and positive predictive values were respectively 87.5%, 98.7%, 87.5%, and 98.7% for PCT and 50%, 97%, 67%, and 94% for CRP. Serum PCT in cord blood seems to be a useful and early marker of antenatal infection. PMID- 16371392 TI - Withholding and withdrawing of life sustaining treatment in the newborn. AB - The rapid progress of medical technology has resulted in more opportunities to maintain the life of infants in serious and potentially life threatening situations. Whether to treat such infants is a common dilemma. The burden of these difficult decisions rests almost equally on distraught parents and relatives and on the professional staff of neonatal units. Sometimes, either parents or care teams choose to seek a decision from the courts. Ways of reaching the best possible and most inclusive consensus decisions are examined in this review. PMID- 16371393 TI - Systematic review of the optimal fluid for dilutional exchange transfusion in neonatal polycythaemia. AB - OBJECTIVES: Several studies have shown the efficacy of dilutional exchange transfusion (DET) in reducing haematocrit (Ht) and relieving clinical symptoms in neonatal polycythaemia. We conducted a systematic review to determine the efficacy of crystalloid versus colloid solutions used in DET in an effort to identify the best solution to replace red blood cells. METHODS: The Cochrane Library, MEDLINE, and EMBASE were searched for relevant randomised controlled trials. Quality assessment and data analysis were performed using the methods and software of the Cochrane Collaboration. Relative risk (RR) and weighted mean difference (WMD) were calculated as measures of effect for categorical and continuous outcome data, respectively. Ninety five percent confidence intervals (95% CI) were calculated and a fixed effect model was used for meta-analysis. RESULTS: Six studies with a total of 235 newborns matched our inclusion criteria. When comparing crystalloid and colloid replacement solutions for DET, there was a clinically unimportant difference in Ht at 2-6 h and at 24 h in favour of colloidal solutions (WMD 2.29% (95% CI 1.28 to 3.31) and 1.74% (95% CI 0.80 to 2.68), respectively). This difference in post DET Ht was more evident when normal saline was compared to plasma but absent when normal saline was compared to 5% albumin. CONCLUSION: There is little difference in effectiveness between plasma, 5% albumin, and crystalloid solutions. Since normal saline is cheap, readily available, and does not carry the potential risk of transfusion associated infection, normal saline is the optimal dilutional fluid for exchange transfusion in polycythaemic neonates. PMID- 16371394 TI - Unnatural selection: reducing antibiotic resistance in neonatal units. AB - Antibiotic use selects for antibiotic resistant bacteria. This is an example of rapid Darwinian natural selection in action. It occurs in neonatal intensive care units with the use of parenteral antibiotics, and in the community with oral antibiotic use. A 10 point plan is put forward to reduce antibiotic resistance in neonatal units. PMID- 16371395 TI - Aristotle (384-322 BC): philosopher and scientist of ancient Greece. AB - Aristotle's studies encompassed the entire world of living things. Many of his descriptions and classifications remain sound today. Although not a physician, he exerted a profound influence on medicine for the next 2000 years. PMID- 16371396 TI - Yes, human milk does reduce infection rates in very low birthweight infants. PMID- 16371397 TI - Antenatal maternal transfers in the east of England: role of a centralised cot bureau. PMID- 16371398 TI - Course of symptoms and median nerve conduction values in workers performing repetitive jobs at risk for carpal tunnel syndrome. AB - BACKGROUND: Carpal tunnel syndrome (CTS) is a common occupational disorder associated with repetitive manual work. Little information exists about the possible relation between the variation of biomechanical hand/wrist exposure and the development of symptoms and median nerve conduction values. AIMS: To investigate the prevalence of CTS in a group of workers exposed to intensive use of the hands and the course of symptoms and median nerve conduction values after a period of reduced exposure to biomechanical risk factors. METHODS: CTS was assessed in assembly and non-assembly line workers by means of clinical examinations and nerve conduction studies (NCS). Ergonomic analysis was conducted for each assembly line workstation. RESULTS: Prevalence of CTS was significantly higher in assembly line workers compared to non-assembly line workers but there was a high prevalence of median nerve conduction abnormalities in both groups (60/102 hands and 40/110 hands, respectively). In a sizable proportion of both groups there was no relationship between symptoms and electrodiagnostic findings (45 hands and 48 hands in assembly and non-assembly line workers, respectively). When assembly line workers were re-examined after 2 years following a period of reduced work schedule, a significant proportion reported resolution of symptoms or had reverted to having normal NCS. CONCLUSIONS: In our study, repetitive work was associated with a higher level of CTS and abnormal NCS. These findings appeared to be reversible following a period of less repetitive work. Overall, there was generally poor correlation between symptoms and electrodiagnostic findings. PMID- 16371399 TI - Medical students' evaluation of a teaching session in occupational medicine: the value of a workplace visit. AB - BACKGROUND: Undergraduate teaching in occupational medicine in UK medical schools is in decline. We developed a teaching programme for the new curriculum at our medical school, and then used student evaluation to tailor it to students' perceived learning needs. AIMS: To examine medical students' perception of a teaching session in occupational medicine after changes made in the light of earlier student evaluation, and in particular their views on the inclusion of a workplace visit. METHODS: Questionnaire evaluation feedback completed immediately after teaching sessions, triangulated with a student focus group session conducted by an external facilitator to explore what students valued most and least and why. Comparison of student evaluations before and after changes introduced in the light of student evaluations. RESULTS: Students' perception of the usefulness and relevance of the session significantly improved after the changes. Students consistently identified the use of case scenarios as helpful but demonstrated a dichotomy of opinion about the value of a workplace visit. Overall, students valued the brevity of the session that resulted from removing the workplace visit. CONCLUSIONS: It is possible to enhance students' perception of the value of a teaching session by modifying the session in the light of student-based evaluation. PMID- 16371400 TI - Job strain and neck-shoulder symptoms: a prevalence study of women and men white collar workers. AB - BACKGROUND: Neck-shoulder symptoms are frequent among workers. Psychosocial factors at work have been associated with neck-shoulder symptoms, but few studies have examined job strain, the combined effect of high psychological demands (PD) and low decision latitude (DL). AIMS: To examine the association between psychosocial factors at work and the prevalence of self-reported neck-shoulder symptoms among white-collar workers. METHODS: In a cross-sectional study of 1543 white-collar workers, PD and DL at work were measured with Karasek's questionnaire. Prevalent cases were workers for whom neck-shoulder symptoms were present for >or=3 days during the previous 7 days and for whom pain intensity was greater than half the visual analogue scale. Gender and social support at work were evaluated as potential effect modifiers. RESULTS: Workers exposed to high job strain had a higher prevalence of neck-shoulder symptoms [adjusted prevalence ratio (PR): 1.54, 95% confidence interval (CI): 1.00-2.37]. No modifying effect of gender was observed in this association. The effect of job strain was stronger in workers with low social support (adjusted PR: 1.84, 95% CI: 0.92-3.68). These associations tended to be stronger and/or more precise when using alternative exposures and case definition. Namely, a stronger job strain effect was observed when a tertile cut-off was used to classify exposure (adjusted PR: 2.47, 95% CI: 1.15-5.32). CONCLUSION: These results suggest that primary prevention of neck shoulder symptoms among white-collar workers should consider the exposure to job strain, especially when workers are exposed to low social support at work. PMID- 16371401 TI - Specification of adaxial and abaxial stomata, epidermal structure and photosynthesis to CO2 enrichment in maize leaves. AB - Acclimation to CO2 enrichment was studied in maize plants grown to maturity in either 350 or 700 microl l-1 CO2. Plants grown with CO2 enrichment were significantly taller than those grown at 350 microl l-1 CO2 but they had the same number of leaves. High CO2 concentration led to a marked decrease in whole leaf chlorophyll and protein. The ratio of stomata on the adaxial and abaxial leaf surfaces was similar in all growth conditions, but the stomatal index was considerably increased in plants grown at 700 microl l-1 CO2. Doubling the atmospheric CO2 content altered epidermal cell size leading to fewer, much larger cells on both leaf surfaces. The photosynthesis and transpiration rates were always higher on the abaxial surface than the adaxial surface. CO2 uptake rates increased as atmospheric CO2 was increased up to the growth concentrations on both leaf surfaces. Above these values, CO2 uptake on the abaxial surface was either stable or increased as CO2 concentration increased. In marked contrast, CO2 uptake rates on the adaxial surface were progressively inhibited at concentrations above the growth CO2 value, whether light was supplied directly to this or the abaxial surface. These results show that maize leaves adjust their stomatal densities through changes in epidermal cell numbers rather than stomatal numbers. Moreover, the CO2-response curve of photosynthesis on the adaxial surface is specifically determined by growth CO2 abundance and tracks transpiration. Conversely, photosynthesis on the abaxial surface is largely independent of CO2 concentration and rather independent of stomatal function. PMID- 16371402 TI - Phenotypic plasticity and growth temperature: understanding interspecific variability. AB - The subject of this review is the impact of long-term changes in temperature on plant growth and its underlying components. The discussion highlights the extent to which thermal acclimation of metabolism is intrinsically linked to the plasticity of a range of biochemical and morphological traits. The fact that there is often a trade-off between temperature-mediated changes in net assimilation rates (NAR) and biomass allocation [in particular the specific leaf area (SLA)] when plants are grown at different temperatures is also highlighted. Also discussed is the role of temperature-mediated changes in photosynthesis and respiration in determining NAR values. It is shown that in comparisons that do not take phylogeny into account, fast-growing species exhibit greater temperature dependent changes in RGR, SLA, and NAR than slow-growing plants. For RGR and NAR, such trends are maintained within phylogenetically independent contrasts (i.e. species adapted to more-favourable habitats consistently exhibit greater temperature-mediated changes than their congeneric counterparts adapted to less favourable habitats). By contrast, SLA was not consistently more thermally plastic in species from favourable habitats. Interestingly, biomass allocation between leaves and roots was consistently more plastic in slow-growing species within individual phylogenetically independent contrasts, when plants were grown under contrasting temperatures. Finally, how interspecific variations in NAR account for an increasing proportion of variability in RGR as growth temperatures decrease is highlighted. Conversely, SLA played a more dominant role in determining interspecific variability in RGR at higher growth temperatures; thus, the importance of SLA in determining interspecific variation in RGR could potentially increase if annual mean temperatures increase in the future. PMID- 16371403 TI - Excess risk of fatal coronary heart disease associated with diabetes in men and women: meta-analysis of 37 prospective cohort studies. AB - OBJECTIVE: To estimate the relative risk for fatal coronary heart disease associated with diabetes in men and women. DESIGN: Meta-analysis of prospective cohort studies. DATA SOURCES: Studies published between 1966 and March 2005, identified through Embase and Medline, using a combined text word and MESH heading search strategy, in addition to studies from the Asia Pacific Cohort Studies Collaboration. REVIEW METHODS: Studies were eligible if they had reported estimates of the relative risk for fatal coronary heart disease comparing men and women with and without diabetes. Studies were excluded if the estimates were not adjusted at least for age. RESULTS: 37 studies of type 2 diabetes and fatal coronary heart disease among a total of 447,064 patients were identified. The rate of fatal coronary heart disease was higher in patients with diabetes than in those without (5.4 v 1.6%). The overall summary relative risk for fatal coronary heart disease in patients with diabetes compared with no diabetes was significantly greater among women than it was among men: 3.50, 95% confidence interval 2.70 to 4.53 v 2.06, 1.81 to 2.34. After exclusion of the eight studies that had adjusted only for age, the difference in risk between the sexes was substantially reduced but still highly significant. The pooled ratio of the relative risks (women: men) from the 29 studies with multiple adjusted estimates was 1.46 (1.14 to 1.88). CONCLUSIONS: The relative risk for fatal coronary heart disease associated with diabetes is 50% higher in women than it is in men. This greater excess coronary risk may be explained by more adverse cardiovascular risk profiles among women with diabetes, combined with possible disparities in treatment that favour men. PMID- 16371404 TI - Low-density lipoprotein size and cardiovascular risk assessment. AB - A predominance of small, dense low-density lipoproteins (LDL) has been accepted as an emerging cardiovascular risk factor by the National Cholesterol Education Program Adult Treatment Panel III. LDL size seems to be an important predictor of cardiovascular events and progression of coronary heart disease and evidences suggests that both quality (particularly small, dense LDL) and quantity may increase cardiovascular risk. However, other authors have suggested that LDL size measurement does not add information beyond that obtained by measuring LDL concentration, triglyceride levels and HDL concentrations. Therefore, it remains debatable whether to measure LDL particle size in cardiovascular risk assessment and, if so, in which categories of patient. Therapeutic modulation of LDL particle size or number appears beneficial in reducing the risk of cardiovascular events, but no clear causal relationship has been shown, because of confounding factors, including lipid and non-lipid variables. Studies are needed to investigate the clinical significance of LDL size measurements in patients with coronary and non-coronary forms of atherosclerosis; in particular, to test whether LDL size is associated with even higher vascular risk, and whether LDL size modification may contribute to secondary prevention in such patients. PMID- 16371406 TI - Multiple sclerosis and risk-taking. PMID- 16371405 TI - Lymphopenia at presentation is associated with increased risk of infections in patients with systemic lupus erythematosus. AB - BACKGROUND: Patients with systemic lupus erythematosus (SLE) frequently suffer from infections, but the predisposing risk factors, as well as the exact frequency and nature of such infections, are not fully understood. AIM: To describe the frequency, types and risk factors for infections in a group of Chinese patients in the early stage of SLE in Hong Kong. DESIGN: Retrospective record study. METHODS: We reviewed the case records of 91 Chinese SLE patients, presenting <12 months after SLE diagnosis. Details of major infections (requiring intravenous antimicrobial therapy, or any confirmed mycobacterial infection) and minor infections were reviewed. Clinical and laboratory features, the systemic lupus erythematosus disease activity index (SLEDAI) at presentation and drug treatment were recorded and analysed. RESULTS: There were 48 major infections and 62 minor infections during 260 patient-years of follow-up. A lymphocyte count < or =1.0 x 10(9)/l at presentation was independently associated with an increased risk for major infection: hazard ratio 4.7 (95%CI 1.6-13.7), p = 0.005. SLEDAI, use of corticosteroids and immunosuppressive therapy were all not associated with increased risk of infection. DISCUSSION: Lymphopenia was an important risk factor for major infections in this group of Chinese patients in the early stages of SLE. SLE patients with lymphopenia at presentation should be closely monitored for the development of infective complications. PMID- 16371407 TI - Availability bias in the management of myocardial infarction. PMID- 16371408 TI - Crusades and mirages. PMID- 16371409 TI - Spatial neglect--a vestibular disorder? AB - The phenomenon of spatial neglect after right brain damage greatly helps our understanding of the normal mechanisms of directing and maintaining spatial attention, of spatial orientation, and the characteristics of neural representation of space. The intriguing symptom is a spontaneous orientation bias towards the right leading to neglect of objects or persons on the left. Interestingly, we observe similar symptoms namely a spontaneous bias of eyes and head along the horizontal dimension of space in patients with unilateral vestibular dysfunction. Further similarities concern anatomical findings. Both spatial neglect and vestibular processing at cortical level show dominance in the right hemisphere and involve common brain areas. Lesion studies in human and monkey, electrical and transcranial magnetic stimulation, as well as functional imaging results have revealed the superior temporal cortex, insula and the temporo-parietal junction to be substantial parts of the multisensory (vestibular) system as well as to be affected in spatial neglect. We argue that these structures are not strictly 'vestibular' but rather have a multimodal character representing a significant site for the neural transformation of converging vestibular, auditory, neck proprioceptive and visual input into higher order spatial representations. Neurons of these regions provide us with redundant information about the position and motion of our body in space. They seem to play an essential role in adjusting body position relative to external space. This view may initiate further development of those strategies to treat spatial neglect that use routes to rehabilitation based on specific manipulations of sensory input feeding into this system. PMID- 16371410 TI - The clinical spectrum of neuralgic amyotrophy in 246 cases. AB - We investigated the symptoms, course and prognosis of neuralgic amyotrophy (NA) in a large group of patients with idiopathic neuralgic amyotrophy (INA, n = 199) and hereditary neuralgic amyotrophy (HNA, n = 47) to gain more insight into the broad clinical spectrum of the disorder. Several findings from earlier smaller scale studies were tested, and for the first time the potential differences between the hereditary and idiopathic phenotypes and between males and females were explored. Generally, the course of the pain manifests itself in three consecutive phases with an initial severe, continuous pain lasting for approximately 4 weeks on average. Sensory involvement was quite common and found in 78.4% of patients but was clinically less impairing than the initial pain and subsequent paresis. As a typically patchy disorder NA can affect almost any nerve in the brachial plexus, although damage in the upper and middle trunk distribution with involvement of the long thoracic and/or suprascapular nerve occurred most frequently (71.1%). We found no correlation between the distribution of motor and sensory symptoms. In INA recurrent attacks were found in 26.1% of the patients during an average 6 year follow-up. HNA patients had an earlier onset (28.4 versus 41.3 years), more attacks (mean 3.5 versus 1.5) and more frequent involvement of nerves outside the brachial plexus (55.8 versus 17.3%) than INA patients, and a more severe maximum paresis, with a subsequent poorer functional outcome. In males the initial pain tended to last longer than it did in females (45 versus 23 days). In females the middle or lower parts of the brachial plexus were involved more frequently (23.1 versus 10.5% in males), and their functional outcome was worse. Overall recovery was less favourable than usually assumed, with persisting pain and paresis in approximately two-thirds of the patients who were followed for 3 years or more. PMID- 16371411 TI - Randomized phase III study of 3-weekly versus weekly docetaxel in pretreated advanced non-small-cell lung cancer: a Spanish Lung Cancer Group trial. AB - BACKGROUND: Docetaxel is a widely accepted second-line treatment in advanced non small-cell lung cancer (NSCLC) with a risk of myelotoxicity. This study evaluated the efficacy and toxicity profile of two docetaxel regimens in NSCLC patients who had failed first-line non-docetaxel-based chemotherapy. PATIENTS AND METHODS: A total of 259 patients from 33 Spanish centers were randomized to receive either docetaxel 75 mg/m(2) administered every 3 weeks (3W arm) or docetaxel 36 mg/m(2) given weekly (1W arm) for 6 weeks followed by 2 weeks of rest. The primary end point was 1-year survival; secondary end points were median survival, time to progression, response and toxicity. RESULTS: One-year survival was 27% in the 3W and 22% in the 1W arm. Median time to progression was also similar in the two arms. Median survival was 6.6 months in the 3W arm versus 5.4 months in the 1W arm (P = 0.075). Response rates were 9.3% in the 3W arm and 4.8% in the 1W arm. More patients in the 1W arm experienced mucositis [1W, nine patients (7.2%); 3W, two patients (1.6%); P = 0.032], while febrile neutropenia was significantly higher in the 3W arm [3W, 10 patients (7.8%); 1W, one patient (0.8%); P = 0.010]. CONCLUSIONS: Both weekly and 3-weekly docetaxel were effective and well tolerated, with different toxicity profiles. In general, there was no indication to recommend the weekly schedule. However, the significant lower rate of febrile neutropenia observed in the weekly schedule makes it a good alternative for patients at risk of severe neutropenia. PMID- 16371412 TI - Proceedings of the SMBE Tri-National Young Investigators' Workshop 2005. Investigating the intron recognition mechanism in eukaryotes. AB - Recent studies indicate that many introns, as well as the complex spliceosomal mechanism to remove them, were present early in eukaryotic evolution. This study examines intron and exon characteristics from annotations of whole genomes to investigate the intron recognition mechanism. Exon definition uses the exon as the unit of recognition, placing length constraints on the exon but not on the intron (allowing it a greater range of lengths). In contrast, intron definition uses the intron itself as the unit of recognition and thus removes constraints on internal exon length forced by the use of an exon definition mechanism. Thus, intron and exon lengths within a genome can reflect the constraints imposed by its splicing. This study shows that it is possible firstly to recover valid intron and exon information from genome annotation. We then compare internal intron and exon information from a range of eukaryotic genomes and investigate possible evolutionary length constraints on introns and exons and how they can impact on the intron recognition mechanism. Results indicate that exon definition based mechanisms may predominate in vertebrates although the exact system in fish is expected to show some differences with the better characterized system from mammals. We also raise the possibility that the last common ancestor of plants and animals contained some type of exon definition and that this mechanism was replaced in some genes and lineages by intron definition, possibly as a result of intron loss and/or intron shortening. PMID- 16371413 TI - Metamodels of bias in Cox proportional-hazards and logistic regressions with heteroscedastic measurement error under group-level exposure assessment. AB - In occupational epidemiology, group-based exposure assessment entails estimating the average exposure level in a group of workers and assigning the average to all members of the group. The assigned exposure values can be used in epidemiological analyses and have been shown to produce virtually unbiased relative-risk estimates in many situations. Although the group-based exposure assessment continues to be used widely, it is unclear whether it produces unbiased relative risk estimates in all circumstance, specifically in Cox proportional-hazards and logistic regressions when between-worker variance is not constant but proportional to the true group mean. This question is important because (i) between-worker variance has been shown to differ among exposure groups in occupational epidemiological studies and (ii) recent theoretical work has suggested that bias may exist in such situations. We conducted computer simulations of occupational epidemiological studies to address this question and analysed simulation results using 'metamodelling'. The results indicate that small-to-negligible bias can be expected to result from heteroscedastic between worker variance. Cox proportional-hazards models can produce attenuated risk estimates, while logistic regression may result in overestimation of risk gradient. Bias caused by ignoring the heteroscedastic measurement error is unlikely to be large enough to alter the conclusion about the direction of exposure-disease association in occupational epidemiology. PMID- 16371414 TI - Statistical modelling of formaldehyde occupational exposure levels in French industries, 1986-2003. AB - Occupational exposure databanks (OEDBs) have been cited as sources of exposure data for exposure surveillance and exposure assessment in epidemiology. In 2003, an extract was made from COLCHIC, the French national OEDB, of all concentrations of formaldehyde. The data were analysed with extended linear mixed-effects models in order to identify influent variables and elaborate a multi-sector picture of formaldehyde exposures. Respectively, 1401 and 1448 personal and area concentrations were available for the analysis. The fixed effects of the personal and area models explained, respectively, 57 and 53% of the total variance. Personal concentrations were related to the sampling duration (short-term higher than TWA levels), decreased with the year of sampling (-9% per year) and were higher when local exhaust ventilation was present. Personal levels taken during planned visits and for occupational illness notification purpose were consistently lower than those taken during ventilation modification programmes or because the hygienist suspected the presence of significant risk or exposure. Area concentrations were related to the sampling duration (short-term higher than TWA levels), and decreased with the year of sampling (-7% per year) and when the measurement sampling flow increased. Significant within-facility (correlation coefficient 0.4-0.5) and within-sampling campaign correlation (correlation coefficient 0.8) was found for both area and personal data. The industry/task classification appeared to have the greatest influence on exposure variability while the sample duration and the sampling flow were significant in some cases. Estimates made from the models for year 2002 showed elevated formaldehyde exposure in the fields of anatomopathological and biological analyses, operation of gluing machinery in the wood industry, operation and monitoring of mixers in the pharmaceutical industry, and garages and warehouses in urban transit authorities. PMID- 16371415 TI - Benzene exposure on a crude oil production vessel. AB - OBJECTIVES: The aim was to describe the personal exposure to benzene on a typical crude oil production vessel and to identify factors influencing the exposure level. METHODS: The study population included process operators, deck workers, mechanics and contractors on a production vessel in the Norwegian sector of the North Sea. The personal exposure to benzene during ordinary activity, during a short shutdown and during tank work was monitored using organic vapour passive dosimeter badges (3M 3500). Information on the tasks performed on the day of sampling was recorded. Exposure was assessed by grouping the measurements according to job category, mode of operation and the tasks performed on the sampling day. Univariate analysis of variance was used to test the differences between the groups. RESULTS: Forty-two workers participated in the exposure assessment, comprising a total of 139 measurements. The arithmetic and geometric mean of benzene exposure for all measurements was 0.43 and 0.02 p.p.m., respectively. Twenty-five measurements (18%) were below the limit of detection (0.001 p.p.m.), while ten samples (7%) exceeded the occupational exposure limit of 0.6 p.p.m. The geometric mean exposure was 0.004 p.p.m. (95% CI 0.003-0.006) during ordinary activity, 0.01 p.p.m. (95% CI 0.005-0.02) during shutdown and 0.28 p.p.m. (95% CI 0.16-0.49) during tank work. Workers performing annual cleaning and maintenance of tanks containing crude oil or residues of crude oil had higher levels of exposure than workers performing other tasks, including work near open hydrocarbon-transport systems (all P < 0.001). However, because of the mandatory use of respirators, the actual personal benzene exposure was lower. The job categories explained only 5% of the variance in exposure, whereas grouping by mode of operation explained 54% of the variance and grouping by task 68%. CONCLUSION: The results show that, although benzene exposure during ordinary and high activity seems to be low in the processing area on the production vessel, cleaning of tanks and performing maintenance work in a cleaned tank have a potential for high exposure. PMID- 16371416 TI - Lung fibre burden in lung cancer cases employed in the rock and slag wool industry. AB - OBJECTIVES: To evaluate the relationship between estimated exposure to man-made vitreous fibres (MMVF) and to asbestos fibres and their concentration in the lung tissue of lung cancer cases amongst MMVF production workers. METHODS: Retrospective retrieval of available lung tissue specimens was conducted following a case-control study that assessed estimated occupational exposures of MMVF workers. Fibre recovery and analysis by transmission electron microscopy (TEM) were conducted to determine fibre type, fibre dimension and numbers per gram of dry lung tissue. For cases with detailed exposure data, geometric mean (GM) concentrations were compared across the exposure categories, and regression models were used to investigate the relationship between the lung fibres and the variables of estimated exposure, with and without additional variables that may affect fibre retention. RESULTS: A total of 24 samples from 17 cases of lung cancer were available for analysis: MMVF were detected in all cases. Asbestos fibres were detected in 16. No difference or trend in GM MMVF concentration was observed across the estimated exposure categories. Odds ratio (OR) for MMVF g(-1) dry lung was 0.5 (95% confidence interval: 0.1-2.4) for the second, and 3.5 (0.6 18.9) for the third quartile of index of average exposure to MMVF in industry, compared with the first (lowest exposed) quartile (no cases in the highest quartile). CONCLUSIONS: No observable relationship existed between estimated exposure and directly-measured lung fibres among this sample of cases. Retrospective specimen collection, intra-individual variability in fibre concentration, effect of unknown factors on fibre retention and small sample size militated against this study providing evidence for or against a relationship between estimated exposure and lung fibre concentrations. PMID- 16371417 TI - Identification of markers for PCB exposure in plasma from Swedish construction workers removing old elastic sealants. AB - The objective of the present study was to identify PCB-indicators of occupational exposure related to the removal of old elastic sealants. Blood samples were collected from workers involved in removing elastic sealants. Samples were also taken from age- and sex-matched controls. A majority of the exposed workers were re-sampled after 10 months. All samples were analysed for 19 PCBs. The levels in the exposed workers were twice as high as those in the controls, 575 and 267 ng g(-1) lipid, respectively, but were essentially unchanged at the second sampling. The PCB patterns also differed. Levels of many less chlorinated PCBs were much higher in the exposed workers, compared to the controls, and principal component analysis (PCA) revealed that easily metabolized PCBs decreased in the exposed workers during the study. This finding indicates that elimination exceeded uptake during the study period, and that the safety information given to the workers had been effective. PCA was also used to identify exposure markers. The relatively persistent PCBs 56/60 and 66, the easily metabolized PCBs 44, 70 and 110 (with vicinal hydrogens in meta/para-positions) and the very persistent PCBs 153 and 180 were found to be good markers for occupational, recent occupational and background (dietary) exposure, respectively. A PCA model based on these markers was equally effective in differentiating between exposed individuals and controls, and between recent and less recent exposure, as a model based on all PCBs. PMID- 16371418 TI - Comments about the proposal to adapt the WASP programme to fibre counting tests. PMID- 16371419 TI - Estimating the number of asbestos-related cancer deaths in Great Britain. PMID- 16371420 TI - Identification of gene expression signatures in autoimmune disease without the influence of familial resemblance. AB - Even though autoimmune diseases are heterogeneous, believed to result from the interaction between genetic and environmental components, patients with these disorders exhibit reproducible patterns of gene expression in their peripheral blood mononuclear cells. A portion of this gene expression profile is a property of familial resemblance rather than autoimmune disease. Here, we wanted to identify the portion of this gene expression profile that is independent of familial resemblance and determine whether it is a product of disease duration, disease onset or other factors. By employing supervised clustering algorithms, we identified 100 genes whose expression profiles are shared in individuals with various autoimmune diseases but are not shared by unaffected family members of individuals with autoimmune disease or by controls. Individuals with early disease (1 year after onset) and established disease (10 years after onset) exhibit a near-identical expression pattern, suggesting that this unique profile is a product of disease onset rather than disease duration. PMID- 16371421 TI - Integrative genetic analysis of transcription modules: towards filling the gap between genetic loci and inherited traits. AB - Genetic loci that regulate inherited traits are routinely identified using quantitative trait locus (QTL) mapping methods. However, the genotype-phenotype associations do not provide information on the gene expression program through which the genetic loci regulate the traits. Transcription modules are 'self consistent regulatory units' and are closely related to the modular components of gene regulatory network [Ihmels, J., Friedlander, G., Bergmann, S., Sarig, O., Ziv, Y. and Barkai, N. (2002) Revealing modular organization in the yeast transcriptional network. Nat. Genet., 31, 370-377; Segal, E., Shapira, M., Regev, A., Pe'er, D., Botstein, D., Koller, D. and Friedman, N. (2003) Module networks: identifying regulatory modules and their condition-specific regulators from gene expression data. Nat. Genet., 34, 166-176]. We used genome-wide genotype and gene expression data of a genetic reference population that consists of mice of 32 recombinant inbred strains to identify the transcription modules and the genetic loci regulating them. Twenty-nine transcription modules defined by genetic variations were identified. Statistically significant associations between the transcription modules and 18 classical physiological and behavioral traits were found. Genome-wide interval mapping showed that major QTLs regulating the transcription modules are often co-localized with the QTLs regulating the associated classical traits. The association and the possible co-regulation of the classical trait and transcription module indicate that the transcription module may be involved in the gene pathways connecting the QTL and the classical trait. Our results show that a transcription module may associate with multiple seemingly unrelated classical traits and a classical trait may associate with different modules. Literature mining results provided strong independent evidences for the relations among genes of the transcription modules, genes in the regions of the QTLs regulating the transcription modules and the keywords representing the classical traits. PMID- 16371422 TI - Mitochondrial iron detoxification is a primary function of frataxin that limits oxidative damage and preserves cell longevity. AB - Friedreich ataxia is a severe autosomal-recessive disease characterized by neurodegeneration, cardiomyopathy and diabetes, resulting from reduced synthesis of the mitochondrial protein frataxin. Although frataxin is ubiquitously expressed, frataxin deficiency leads to a selective loss of dorsal root ganglia neurons, cardiomyocytes and pancreatic beta cells. How frataxin normally promotes survival of these particular cells is the subject of intense debate. The predominant view is that frataxin sustains mitochondrial energy production and other cellular functions by providing iron for heme synthesis and iron-sulfur cluster (ISC) assembly and repair. We have proposed that frataxin not only promotes the biogenesis of iron-containing enzymes, but also detoxifies surplus iron thereby affording a critical anti-oxidant mechanism. These two functions have been difficult to tease apart, however, and the physiologic role of iron detoxification by frataxin has not yet been demonstrated in vivo. Here, we describe mutations that specifically impair the ferroxidation or mineralization activity of yeast frataxin, which are necessary for iron detoxification but do not affect the iron chaperone function of the protein. These mutations increase the sensitivity of yeast cells to oxidative stress, shortening chronological life span and precluding survival in the absence of the anti-oxidant enzyme superoxide dismutase. Thus, the role of frataxin is not limited to promoting ISC assembly or heme synthesis. Iron detoxification is another function of frataxin relevant to anti-oxidant defense and cell longevity that could play a critical role in the metabolically demanding environment of non-dividing neuronal, cardiac and pancreatic beta cells. PMID- 16371423 TI - Deleterious and protective properties of an aggregate-prone protein with a polyalanine expansion. AB - Many aggregate-prone proteins, including proteins with long polyglutamine or polyalanine tracts, cause human diseases. Polyalanine proteins may also be present in the tissue of polyglutamine diseases as a result of frameshifting of the primary polyglutamine-encoding (CAG)n repeat mutation. We have generated a Drosophila model expressing green fluorescent protein tagged to 37 alanines that manifests both toxicity and inclusion formation in various tissues. Surprisingly, we show that this aggregate-prone protein with a polyalanine expansion can also protect against polyglutamine toxicity, which can be explained by induction of heat-shock response. A heat-shock response was also seen in an oculopharyngeal muscular dystrophy mouse model expressing an authentic polyalanine-expanded protein. We also show that long polyalanines can protect against a pro-apoptotic stimulus or the toxicity caused by the long polyalanines themselves. Thus, overexpression of an aggregate-prone protein without any normal functions can result in both pathogenic and protective effects in cell culture and in vivo. PMID- 16371424 TI - Mathematically modelling the effects of counting factor in Dictyostelium discoideum. AB - Size regulation is a crucial feature in many biological systems, with misregulation leading to dysplasia or hyperplasia. The recent discovery of counting factor (CF) in Dictyostelium discoideum will lead to a greater understanding of how the system regulates the size of a group of cells. In this paper we mathematically model the known effects of CF using two different models: a cellular automata model and a discrete continuum hybrid model. With the use of these models we are able to understand how modulation of adhesion and motile forces by CF can facilitate stream breakup. In addition, the modelling suggests a new possible mechanism for stream breakup involving the frequency of cell reorientation. PMID- 16371425 TI - Essential role for the Menkes ATPase in activation of extracellular superoxide dismutase: implication for vascular oxidative stress. AB - Extracellular superoxide dismutase (SOD3), a secretory copper enzyme, plays an important role in atherosclerosis and hypertension by modulating the levels of extracellular superoxide anion (O2*-) in the vasculature. Little is known about the mechanisms by which SOD3 obtains its catalytic copper cofactor. Menkes ATPase (MNK) has been shown to transport cytosolic copper to the secretory pathway in nonvascular cells. We performed the present study to determine whether MNK is required for the activation of SOD3 in the vasculature. Here we show that MNK was highly expressed in the various vascular tissues and cells. Aortas and cultured fibroblasts from MNK mutant (MNK(mut)) mice showed a marked decrease in specific activity of SOD3, but not SOD1 (cytosolic form), which was partially restored by copper addition. Copper treatment in wild-type cells promoted the direct interaction and colocalization of SOD3 with MNK in the trans-Golgi network (TGN), suggesting that MNK transports copper to SOD3 in the TGN. Aortas of MNK(mut) mice revealed a decrease in activity of SOD3, but not SOD1, in association with a robust increase in O2*- levels. Finally, both MNK and SOD3 proteins were highly expressed in the intimal lesions of atherosclerotic vessels. In conclusion, vascular MNK plays an essential role in full activity of SOD3 through transporting copper to SOD3 in the TGN, thereby regulating O2*- levels in the vasculature. These studies provide a novel insight into vascular MNK as a critical modulator of "superoxide" stress, which may contribute to cardiovascular disease. PMID- 16371426 TI - Impairment of the ubiquitin-proteasome system in desminopathy mouse hearts. AB - Protein misfolding and aberrant aggregation are associated with many severe disorders, such as neural degenerative diseases, desmin-related myopathy (DRM), and congestive heart failure. Intrasarcoplasmic amyloidosis and increased ubiquitinated proteins are observed in human failing hearts. The pathogenic roles of these derangements in the heart remain unknown. The ubiquitin-proteasome system (UPS) plays a central role in intracellular proteolysis and regulates critical cellular processes. In cultured cells, aberrant aggregation by a mutant (MT) or misfolded protein impairs the UPS. However, this has not been demonstrated in intact animals, and it is unclear how the UPS is impaired. Cross breeding UPS reporter mice with a transgenic mouse model of DRM featured by aberrant protein aggregation in cardiomyocytes, we found that overexpression of MT-desmin but not normal desmin protein impairs UPS proteolytic function in the heart. The primary defect does not appear to be in the ubiquitination or the proteolytic activity of the 20S proteasome, because ubiquitinated proteins and the peptidase activities of 20S proteasomes were significantly increased rather than decreased in the DRM heart. Therefore, the defect resides apparently in the entry of ubiquitinated proteins into the 20S proteasome. Consistent with this notion, key components (Rpt3 and Rpt5) of 19S proteasomes were markedly decreased, while major components of 20S proteasomes were increased. Additional experiments with HEK cells suggest that proteasomal malfunction observed in MT desmin hearts is not secondary to cardiac malfunction or to disruption of desmin filaments. Thus, UPS impairment may represent an important pathogenic mechanism underlying cardiac disorders with abnormal protein aggregation. PMID- 16371427 TI - Enhanced programmed cell death of iron-deficient erythrocytes. AB - Exposure of erythrocytes to osmotic shock, oxidative stress, and energy depletion activates Cl--sensitive and Ca2+-permeable cation channels. Subsequent Ca2+ entry triggers eryptosis, characterized by erythrocyte shrinkage, membrane blebbing, and phosphatidylserine exposure all features typical for apoptotic death of nucleated cells. Erythrocytes exposing phosphatidylserine are recognized, bound, engulfed, and degraded by macrophages. Eryptosis thus fosters clearance of affected erythrocytes from circulating blood. Iron deficiency leads to anemia, in part by decreasing erythrocyte life span. In this study, phosphatidylserine exposure, cell size, and cytosolic Ca2+ were measured by FACS analysis of annexin V binding, forward scatter, and Fluo-3 fluorescence, respectively. Erythrocytes from mice on control diet were compared with erythrocytes from mice exposed 10 weeks to iron-deficient diet. Iron deficiency significantly (P<0.001) enhanced erythrocyte annexin-V binding (from 2.4 to 3.7%), decreased forward scatter (from 544 to 393), and increased cytosolic Ca2+ concentration. 45Ca2+ flux measurements and patch clamp experiments revealed enhanced Ca2+ uptake (by 2.3-fold) and cation channel activity. The half-life of fluorescence-labeled, iron-deficient, or Ca2+-loaded erythrocytes was significantly reduced compared with control erythrocytes. Thus, the experiments reveal a novel mechanism triggered by iron deficiency, which presumably contributes to accelerated clearance of erythrocytes in iron deficiency anemia. PMID- 16371428 TI - Mechanisms of cell death and neuroprotection by poloxamer 188 after mechanical trauma. AB - The mechanisms of cell death and the progressive degeneration of neural tissue following traumatic brain injury (TBI) have come under intense investigation. However, the complex interactions among the evolving pathologies in multiple cell types obscure the causal relationships between the initial effects of the mechanical trauma at the cellular level and the long-term dysfunction and neuronal death. We used an in vitro model of neuronal injury to study the mechanisms of cell death in response to a well-defined mechanical insult and found that the majority of dead cells were apoptotic. We have previously reported that promotion of membrane repair acutely with the non-ionic surfactant poloxamer 188 (P188) restored cell viability to control values at 24 h postinjury. Here, we showed that P188 significantly inhibits apoptosis and prevents necrosis. We also examined the role of mitogen-activated protein kinases (MAPKs) in cell death. There was a rapid, transient activation of extracellular signal-regulated kinases, c-Jun N-terminal kinase, and p38s after mechanical insult. Of these, activation of the proapoptotic p38 was the greatest. Treatment with P188 inhibited p38 activation; however, direct inhibition of p38 by SB203580, which selectively inhibits the activity of the p38 MAPK, provided only partial inhibition of apoptosis and had no effect on necrosis. These data suggest that multiple signaling pathways may be involved in the long-term response of neurons to mechanical injury. Furthermore, that the membrane resealing action of P188 provides such significant protection from both necrosis and apoptosis suggests that acute membrane damage due to trauma is a critical precipitating event that is upstream of the many signaling cascades contributing to the subsequent pathology. PMID- 16371429 TI - Functional recovery of peripheral blood mononuclear cells in modeled microgravity. AB - Immune function is suppressed in space flight, demonstrated by reduced mitogen stimulated proliferation of postflight astronaut peripheral blood mononuclear cells (PBMCs). While flight studies are limited, the development of rotating wall vessel (RWV) bioreactors, such as the high aspect ratio vessel (HARV), has facilitated ground-based studies of the effects of modeled microgravity (MMG) on cell-mediated immunity. Astronauts regain immune function 3 days postflight, but this recovery has not yet been demonstrated following MMG. MMG eliminated phytohemagglutinin (PHA)-stimulated proliferation of PBMCs. Upon removal from HARV, full recovery was gradually achieved over a 72 h period, in agreement with postflight studies of astronauts. Recovery from MMG delayed, but did not reduce, the maximal proliferative response compared with PHA-activated stationary cultures. Likewise, peak expression of T cell surface activation markers CD69 and CD25 was delayed upon stimulation following exposure to MMG. MMG and recovery from MMG differentially affected the detection of IL-2 and IFN in supernatants. Further development of this model of immune recovery is important for investigating the mechanisms of immune suppression and recovery in space flight, as well as possible countermeasures to prevent immunosuppression or enhance recovery. Given the analogous immune suppression observed in microgravity, MMG, and aging, further investigation may also lead to advances in anti-aging medicine. PMID- 16371430 TI - Renal phenotypes related to hepatocyte nuclear factor-1beta (TCF2) mutations in a pediatric cohort. AB - The hepatocyte nuclear factor-1beta encoded by the TCF2 gene plays a role for the specific regulation of gene expression in various tissues such as liver, kidney, intestine, and pancreatic islets and is involved in the embryonic development of these organs. TCF2 mutations are known to be responsible for the maturity-onset diabetes of the young type 5 associated with renal manifestations. Several observations have suggested that TCF2 mutations may be involved in restricted renal phenotypes. Eighty children (median age at diagnosis 0.2 yr) with renal cysts, hyperechogenicity, hypoplasia, or single kidneys were studied. Quantitative multiplex PCR amplification of short fluorescence fragments for the search of large genomic rearrangements and sequencing for the detection of point mutations were performed. TCF2 anomalies were detected in one third of patients (25 of 80). The main alteration was the complete deletion of the TCF2 gene detected in 16 patients. Family screening revealed de novo TCF2 anomalies in nine of 17 probands with a high prevalence of deletions (seven of nine). TCF2 anomalies were associated with bilateral renal anomalies (P < 0.001) and bilateral cortical cysts (P < 0.001). However, abnormal renal function, detected in 40% of patients, was independent of the TCF2 genotype. No difference in renal function or severity of renal morphologic lesions was observed between patients with a TCF2 deletion and those with point mutations. In conclusion, TCF2 molecular anomalies are involved in restricted renal phenotype in childhood without alteration of glucose metabolism. These findings have important implications in the diagnosis of patients with renal dysplasia with cysts and their follow-up. PMID- 16371432 TI - Antiatherogenic effects of angiotensin receptor antagonism in mild renal dysfunction. AB - Angiotensin II (Ang II) increases atherosclerotic cardiovascular disease. Renal damage that is characterized by activation of Ang II markedly potentiates the risk for atherosclerosis, even in the setting of subtle renal impairment. Therefore, whether antagonism of Ang II actions can modify atherosclerosis in a model of mild renal impairment was examined. Apolipoprotein E-deficient spontaneously hyperlipidemic mice underwent uninephrectomy (UNx) or sham operation (sham) followed by treatment with Ang II receptor antagonist losartan or hydralazine for 12 wk. While UNx did not increase the serum creatinine levels, BP and lipids were higher in UNx mice than in age-matched sham controls with intact kidneys. UNx caused a dramatic increase in the extent and the number of atherosclerotic lesions together with greater macrophage-positive area and more disruption in the elastin component of the extracellular matrix versus sham. Ang II antagonism dramatically decreased the UNx-induced acceleration in atherosclerosis in association with decreased macrophage content, linked to decreased macrophage migration in vitro with losartan but not with hydralazine. Aortae of mice treated with Ang II antagonism had fewer elastin breaks together with less immunostaining for the powerful elastolytic enzyme cathepsin S. None of these benefits was observed in the hydralazine-treated mice despite equivalent reduction in BP. These findings support an important role for endogenous Ang II in accelerated atherosclerosis in renal dysfunction and offer a therapeutic intervention with particular benefit in this setting through mechanisms that include reduced vascular macrophage infiltration and preservation of the elastin component of extracellular matrix. PMID- 16371433 TI - Development of gene silencing pyrrole-imidazole polyamide targeting the TGF-beta1 promoter for treatment of progressive renal diseases. AB - Pyrrole-imidazole (Py-Im) polyamides are nuclease-resistant novel compounds that inhibit gene expression by binding to the minor groove of DNA. A Py-Im polyamide that targets rat TGF-beta1 was designed as a gene-silencing agent for progressive renal diseases, and the distribution and the effects of this polyamide on renal injury were examined in Dahl-salt sensitive (Dahl-S) rats. For identification of transcription factor binding elements for activation of the rat TGF-beta1 gene, recombinant TGF-beta1 reporter plasmids were transfected into HEK-293 cells, and promoter activity was measured. Py-Im polyamide was designed to the activator protein-1 binding site of the rat TGF-beta1 promoter. This Py-Im polyamide showed strong, fast, and specific binding to the target DNA in gel mobility shift and Biacore assays. Py-Im polyamide significantly inhibited TGF-beta1 promoter activity and expression of TGF-beta1 mRNA and protein in rat mesangial cells. Intravenously administered fluorescein-labeled polyamide distributed to the kidney of rats. Py-Im polyamide significantly inhibited expression of TGF-beta1 mRNA and protein in the renal cortex of Dahl-S rats and reduced the increase in urinary protein and albumin in Dahl-S rats independent of changes in blood pressure. These results indicate that Py-Im polyamide that targets TGF-beta1 will be a novel gene-silencing agent for the TGF-beta1-associated diseases, including progressive renal diseases. PMID- 16371434 TI - Advanced glycation end products inhibit tubulogenesis and migration of kidney epithelial cells in an ezrin-dependent manner. AB - Nonenzymatic glycation of proteins to form advanced glycation end products (AGE) is implicated in diabetic complications, including nephropathy. It was shown recently that AGE bind to the ERM (ezrin, radixin, and moesin) family of membrane cytoskeletal linker proteins in renal homogenates. Herein is reported the effects of AGE-BSA on ezrin-dependent LLC-PK1 kidney epithelial cellular functions: migration and hepatocyte growth factor (HGF)-induced tubulogenesis. LLC-PK1 cells were stably transfected with cDNA for ezrin sense, ezrin antisense, and N-ezrin. Transfection of LLC-PK1 cells with ezrin antisense and dominant negative N-ezrin decreased basal tubulogenesis and migration relative to vector-only transfection, establishing the ezrin dependency of these processes. AGE-BSA (20 or 40 microM) significantly decreased HGF-induced tubulogenesis and basal migration in two vector control lines relative to BSA-treated cells. However, AGE-BSA inhibition of both HGF-induced tubulogenesis and migration was overcome by overexpressing ezrin. These results demonstrate that the AGE-ezrin interaction significantly alters cellular function. These changes may be relevant to detrimental renal consequences as a result of diabetes. PMID- 16371435 TI - Estimating pediatric glomerular filtration rates in the era of chronic kidney disease staging. AB - With the use of information from a database of pediatric patients with concomitant nuclear GFR and serum creatinine (Cr), estimated GFR equations were derived on the basis of local laboratory methods and population. These formulas then were compared with those recommended by the National Kidney Foundation for estimating GFR in children. For this, their ability to estimate accurately an individual's true GFR and chronic kidney disease stage, identify patients whose true GFR was <60 ml/min per 1.73 m(2), and to identify correctly deterioration in an individual's GFR over time was compared. Next, two methods to estimate GFR in children without the use of height or weight were developed. The first was a height- and weight-independent formula; the second was a novel approach using the Schwartz formula and calculating a Cr cutoff based on age-based estimates of height and GFR level of interest, i.e., <60 ml/min per 1.73 m(2). Our results suggest that if local laboratory constants are derived and a height is known, then the Schwartz formula offers the most accuracy with least mathematical complexity to perform in the clinical setting. If height is not available but the local laboratory constants have been derived, then the British Columbia's Children's Hospital 2 formula is of value; however, in the setting of estimating pediatric renal function in the outpatient laboratory, where neither of these factors is commonly known, an approach whereby a Cr cutoff for a GFR of interest is developed is suggested. Provided are Cr levels that are based on a reference method of Cr measurement to facilitate this approach for the clinician. PMID- 16371436 TI - Ontario pharmacists drop Plan B screening form. PMID- 16371438 TI - Granulocyte-macrophage colony-stimulating factor increases L-arginine transport through the induction of CAT2 in bone marrow-derived macrophages. AB - L-arginine transport is crucial for macrophage activation because it supplies substrate for the key enzymes nitric oxide synthase 2 and arginase I. These enzymes participate in classic and alternative activation of macrophages, respectively. Classic activation of macrophages is induced by type I cytokines, and alternative activation is induced by type II cytokines. The granulocyte macrophage colony-stimulating factor (GM-CSF), in addition to inducing proliferation and differentiation of macrophages, activates arginase I, but its action on L-arginine transport is unknown. We studied the L-arginine transporters that are active in mouse primary bone marrow-derived macrophages (BMM) and examined the effect of GM-CSF treatment on transport activities. Under basal conditions, L-arginine entered mainly through system y(+)L (>75%). The remaining transport was explained by system y(+) (<10%) and a diffusion component (10-15%). In response to GM-CSF treatment, transport activity increased mostly through system y(+) (>10-fold), accounting for about 40% of the total L-arginine transport. The increase in y(+) activity correlated with a rise in cationic amino acid transporter (CAT)-2 mRNA and protein. Furthermore, GM-CSF induced an increase in arginase activity and in the conversion of L-arginine to ornithine, citrulline, glutamate, proline, and polyamines. BMM obtained from CAT2-knockout mice responded to GM-CSF by increasing arginase activity and the expression of CAT1 mRNA, which also encodes system y(+) activity. Nonetheless, the increase in CAT1 activity only partially compensated the lack of CAT2 and L-arginine metabolism was hardly stimulated. We conclude that BMM present mainly y(+)L activity and that, in response to GM-CSF, l-arginine transport augments through CAT2, thereby increasing the availability of this amino acid to the cell. PMID- 16371439 TI - Autocrine loop between TGF-beta1 and IL-1beta through Smad3- and ERK-dependent pathways in rat pancreatic stellate cells. AB - Pancreatic stellate cells (PSCs) are activated during pancreatitis and promote pancreatic fibrosis by producing and secreting ECMs such as collagen and fibronectin. IL-1beta has been assumed to participate in pancreatic fibrosis by activating PSCs. Activated PSCs secrete various cytokines that regulate PSC function. In this study, we have examined IL-1beta secretion from culture activated PSCs as well as its regulatory mechanism. RT-PCR and ELISA have demonstrated that PSCs express IL-1beta mRNA and secrete IL-1beta peptide. Inhibition of TGF-beta(1) activity secreted from PSCs by TGF-beta(1)-neutralizing antibody attenuated IL-1beta secretion from PSCs. Exogenous TGF-beta(1) increased IL-1beta expression and secretion by PSCs in a dose-dependent manner. Adenovirus mediated expression of dominant-negative (dn)Smad2/3 expression reduced both basal and TGF-beta(1)-stimulated IL-1beta expression and secretion by PSCs. Coexpression of Smad3 with dnSmad2/3 restored IL-1beta expression and secretion by PSCs, which were attenuated by dnSmad2/3 expression. In contrast, coexpression of Smad2 with dnSmad2/3 did not alter them. Furthermore, inhibition of IL-1beta activity secreted from PSCs by IL-1beta-neutralizing antibody attenuated TGF beta(1) secretion from PSCs. Exogenous IL-1beta enhanced TGF-beta(1) expression and secretion by PSCs. IL-1beta activated ERK, and PD-98059, a MEK1 inhibitor, blocked IL-1beta enhancement of TGF-beta(1) expression and secretion by PSCs. We propose that an autocrine loop exists between TGF-beta(1) and IL-1beta in activated PSCs through Smad3- and ERK-dependent pathways. PMID- 16371440 TI - Glutamate induces oxidative stress and apoptosis in cerebral vascular endothelial cells: contributions of HO-1 and HO-2 to cytoprotection. AB - In cerebral circulation, epileptic seizures associated with excessive release of the excitatory neurotransmitter glutamate cause endothelial injury. Heme oxygenase (HO), which metabolizes heme to a vasodilator, carbon monoxide (CO), and antioxidants, biliverdin/bilirubin, is highly expressed in cerebral microvessels as a constitutive isoform, HO-2, whereas the inducible form, HO-1, is not detectable. Using cerebral vascular endothelial cells from newborn pigs and HO-2-knockout mice, we addressed the hypotheses that 1) glutamate induces oxidative stress-related endothelial death by apoptosis, and 2) HO-1 and HO-2 are protective against glutamate cytotoxicity. In cerebral endothelial cells, glutamate (0.1-2.0 mM) increased formation of reactive oxygen species, including superoxide radicals, and induced major keystone events of apoptosis, such as NF kappaB nuclear translocation, caspase-3 activation, DNA fragmentation, and cell detachment. Glutamate-induced apoptosis was greatly exacerbated in HO-2 gene deleted murine cerebrovascular endothelial cells and in porcine cells with pharmacologically inhibited HO-2 activity. Glutamate toxicity was prevented by superoxide dismutase, suggesting apoptotic changes are oxidative stress related. When HO-1 was pharmacologically upregulated by cobalt protoporphyrin, apoptotic effects of glutamate in cerebral endothelial cells were completely prevented. Glutamate-induced reactive oxygen species production and apoptosis were blocked by a CO-releasing compound, CORM-A1 (50 microM), and by bilirubin (1 microM), consistent with the antioxidant and cytoprotective roles of the end products of HO activity. We conclude that both HO-1 and HO-2 have anti-apoptotic effects against oxidative stress-related glutamate toxicity in cerebral vascular endothelium. Although HO-1, when induced, provides powerful protection, HO-2 is an essential endogenous anti-apoptotic factor against glutamate toxicity in the cerebral vascular endothelium. PMID- 16371441 TI - Three-dimensional compartmentalization of myosin heavy chain and myosin light chain isoforms in dog thyroarytenoid muscle. AB - The thyroarytenoid muscle, a vocal fold adductor, has important roles in airway protection (e.g., prevention of aspiration) and phonation. Isoform expression of myosin heavy chain (MHC), a major determinant of muscle-shortening velocity, has been reported to be heterogeneous in this muscle in several mammals, differing markedly between the medial and lateral divisions. The objective was to determine the isoform expression patterns of both MHC and myosin light chain (MLC), with the latter having a modulatory role in determining shortening velocity, to further test whether the expression of both myosin subunits differs in multiple specific sites within the divisions of the dog thyroarytenoid muscle, potentially revealing even greater compartmentalization in this muscle. Our results indicate the existence of large gradients in the relative levels of individual MHC isoforms in the craniocaudal axis along the medial layer (i.e., airflow axis), where levels of MHC-I and MHC-IIA are low at both ends of the axis and high in the middle and MHC-IIB has a reciprocal distribution. The lateral layer is more uniform, with high levels of MHC-IIB throughout. The level of MHC-IID is relatively constant along the axis in both layers. Large differences exist in the distribution of MHC isoforms among single fibers isolated from sites along the craniocaudal axis, especially in the lateral layer. Systematic regional variations are apparent in the MLC isoform composition of single fibers as well, including some MLC isoform combinations that are not observed in dog limb muscles. Variations of MHC and MLC isoform expression in the dog thyroarytenoid muscle are greater than previously recognized and suggest an even broader range of contractile properties within this multifunctional muscle. PMID- 16371442 TI - Cytoplasmic targeting signals mediate delivery of phospholemman to the plasma membrane. AB - The FXYD protein family consists of several small, single-span membrane proteins that exhibit a high degree of homology. The best-known members of the family include the gamma-subunit of the Na(+)-K(+)-ATPase and phospholemman (PLM), a phosphoprotein of cardiac sarcolemma. Other members of the family include corticosteroid hormone-induced factor (CHIF), mammary tumor protein of 8 kDa (Mat 8), and related to ion channels (RIC). The exact physiological roles of the FXYD proteins remain unknown. To better characterize the function of the members of the FXYD protein family, we expressed several members of the family in Madin Darby canine kidney (MDCK) cells. All of the FXYD proteins, with the exception of PLM, were primarily found in the basolateral plasma membrane. Surprisingly, PLM, a previously characterized plasma membrane protein, was found to colocalize with the endoplasmic reticulum marker protein disulfide isomerase. Treatment of MDCK cells expressing PLM with an agonist of PKC caused some of the PLM to be redistributed to the plasma membrane. Site-directed mutagenesis of residues within the cytoplasmic domain of PLM indicated that a negative charge at Ser69 is necessary to shift the localization of PLM to the plasma membrane. In addition, other regions of PLM necessary for either its endoplasmic reticulum or plasma membrane localization have been elucidated. In contrast to PLM, the plasma membrane localization of CHIF and RIC was not altered by mutation of potential cytoplasmic phosphorylation sites. Overall, these results suggest that phosphorylation of specific residues of PLM may direct PLM from an intracellular compartment to the plasma membrane. PMID- 16371443 TI - A dynamic analysis of the shade-induced plasticity in Arabidopsis thaliana rosette leaf development reveals new components of the shade-adaptative response. AB - BACKGROUND AND AIMS: It is well known that plant aerial development is affected by light intensity in terms of the date of flowering, the length of stems and petioles, and the final individual leaf area. The aim of the work presented here was to analyse how shade-induced changes in leaf development occur on a dynamic basis from the whole rosette level to that of the cells. METHODS: Care was taken to ensure that light intensity was the only source of micro-meteorological variation in the study. The dynamics of leaf production, rosette expansion, individual leaf area expansion and epidermal cell expansion were analysed in Arabidopsis thaliana plants grown under two light intensities in three independent experiments. KEY RESULTS: The total area of rosette leaves was reduced by the shading treatment. Both the number of leaves produced and their individual leaf areas were reduced. The reduction in leaf number was associated with a reduction in leaf initiation rate and the duration of the phase of leaf production. The reduction in individual leaf area was associated with a reduction in leaf expansion rate and an increase in the duration of leaf expansion. The changes in leaf expansion dynamics were accompanied by a decrease in epidermal cell number which was partly compensated for by an increase in epidermal cell area. Overall, the whole rosette leaf expansion rate was reduced by shading, whereas the total duration of rosette leaf expansion was unaffected. This was mainly due to the accumulation of the increases in the durations of expansion of each individual leaf which was associated with an increase in cell expansion. CONCLUSIONS: The dynamic analysis presented here reveals a new shade-adaptative response mediated via the control of area expansion at the cell, organ and whole plant levels. PMID- 16371444 TI - The pollination ecology of Paraboea rufescens (Gesneriaceae): a buzz-pollinated tropical herb with mirror-image flowers. AB - BACKGROUND AND AIMS: Gesneriaceae is a pantropical plant family with over 3000 species. A great variety of pollination mechanisms have been reported for the neotropical members of the family, but the details of buzz-pollination and enantiostyly for the family have not been described. We investigated the floral biology and pollination ecology of Paraboea rufescens in Xishuangbanna, south west China, considering three aspects: (1) the type of enantiostyly exhibited; (2) whether the species is self-compatible; and (3) whether pollinator behaviour could enhance the precision of pollen transfer between flowers of contrasting stylar orientation. METHODS: Flowering phenology was monitored once a month during vegetative growth, and once a week during flowering both in the field and under cultivation. Pollination manipulations and pollinator observation in the field were conducted. KEY RESULTS: Anthesis occurred early during the morning, and flowers remained open for 1-5 d, depending on weather conditions. Controlled pollinations revealed that P. rufescens is self-compatible, and exhibited inbreeding depression in seed set. Plants were pollinator limited in natural populations. The similar stylar deflection among flowers within a plant limits autonomous self-pollination as well as pollination between flowers. Two species of bumble bees (Bombus spp.), Amegila malaccensis and Nomia sp. effectively pollinated P. rufescens. These pollinators visited flowers in search of pollen with almost the same frequency. None of the pollinators appeared to discriminate between left- or right-handed flowers. CONCLUSIONS: Paraboea rufescens exhibits monomorphic enantiostylous flowers and a buzz-pollination syndrome. Floral morphology in P. rufescens and pollinator foraging behaviour seems likely to reduce self-pollination and pollinations between flowers of the same stylar deflection. PMID- 16371445 TI - Ovary peltate trichomes of Zeyheria montana (Bignoniaceae): developmental ultrastructure and secretion in relation to function. AB - BACKGROUND AND AIMS: Nectar production in the Bignoniaceae species lacking a nectariferous functional disc is ascribed to trichomatic glands around the ovary base and/or on the inner corolla wall. Nevertheless, knowledge about the secretion and function of these glands is very incomplete. The purpose of this paper is to study, from a developmental viewpoint, the ultrastructure, histochemistry and secretory process of the peltate trichomes on the ovary of Zeyheria montana, a species in the Bignoniaceae which has a rudimentary disc. METHODS: Samples of the gynoecium at various developmental stages were fixed and processed for light and electron microscopy. Histochemistry and cytochemistry tests were performed to examine the chemical composition of exudates. Thin layer chromatography was used to determine the presence of alkaloids and terpenes in gynoecium and fruit extracts, and in fresh nectar stored in the nectar chamber. KEY RESULTS: Peltate trichomes at different developmental stages appear side by side from floral budding up to pre-dispersal fruit. Large plastids with an extensive internal membrane system consisting of tubules filled with lipophilic material, abundant smooth endoplasmic reticulum, few Golgi bodies, lipophilic deposits in the smooth endoplasmic reticulum and mitochondria, and scattered cytoplasmic oil droplets are the main characteristics of mature head cells. The secretion which accumulates in the subcuticular space stains positively for hydrophilic and lipophilic substances, with lipids prevailing for fully peltate trichomes. Histochemistry and thin layer chromatography detected terpenes and alkaloids. Fehling's test to detect of sugars in the secretion was negative. CONCLUSIONS: The continuous presence and activity of peltate trichomes on the ovary of Z. montana from early budding through to flowering and fruiting set, and its main chemical components, alkaloids and terpenes, suggest that they serve a protective function and are not related to the floral nectar source or to improving nectar quality. PMID- 16371446 TI - Oxysterol 22(R)-hydroxycholesterol induces the expression of the bile salt export pump through nuclear receptor farsenoid X receptor but not liver X receptor. AB - Oxysterols are intermediates in the synthesis of bile acids and steroid hormones from cholesterol and function as ligands for liver X receptor (LXR). Bile salt export pump (BSEP) is responsible for canalicular secretion of bile acids and is tightly regulated by its substrates bile acids through nuclear receptor farnesoid X receptor (FXR). In a microarray study using human hepatocytes, BSEP was markedly induced not only by chenodeoxycholic acid (CDCA) but also by oxysterol 22(R)-hydroxycholesterol [22(R)-OHC]. We hypothesized that the expression of BSEP was induced by oxysterols through activation of LXR. To test the hypothesis, human primary hepatocytes or hepatoma cells were treated with 22(R)-OHC, and expression of BSEP was determined. The level of BSEP mRNA was increased as much as 5-fold upon oxysterol induction. In contrast to our hypothesis, the oxysterol induced up-regulation of BSEP is mediated through FXR but not LXR. BSEP promoter activity was markedly induced by 22(R)-OHC in the presence of FXR but not LXRs. Mutation of the FXR element IR1 in the BSEP promoter significantly reduced its ability to respond to oxysterol induction. To determine whether 22(R)-OHC and CDCA bind to similar structural features of FXR, site-directed mutagenesis was performed in the FXR ligand binding domain. Mutation of residues R331 and I352 abolished activation mediated by CDCA and 22(R)-OHC. In contrast, substitution of residues L340 and R351 differentiated CDCA- and 22(R)-OHC-mediated activation. In conclusion, oxysterol 22(R)-OHC functions as an FXR ligand to induce BSEP expression and differs in the binding with FXR from CDCA. PMID- 16371447 TI - Inhibition of e-cadherin-mediated homotypic adhesion of Caco-2 cells: a novel evaluation assay for peptide activities in modulating cell-cell adhesion. AB - Transient modulation of E-cadherin-mediated cell-cell adhesion may improve paracellular drug delivery through biological barriers. Therefore, there is a need to develop an efficient method to evaluate cadherin peptides that can modulate the intercellular junctions. The objective of this study was to establish a novel assay to evaluate peptide activity in modulating E-cadherin mediated homophilic interactions, based on the homotypic adhesion of Caco-2 cells. Fluorescence-labeled Caco-2 single cells were incubated with Caco-2 monolayers that were treated beforehand with Ca(2+)-free medium. The homotypic adhesion in the presence or absence of peptide and antibody was determined fluorometrically. The Ca(2+)-deficient pretreatment dramatically increased the number of single cells bound to the monolayers. Immunofluorescence staining showed that some of E-cadherins became accessible without surfactant-induced permeabilization of Caco-2 cell monolayers after the Ca(2+)-deficient pretreatment. The homotypic adhesion was largely dependent on extracellular Ca(2+) concentrations and significantly inhibited by the presence of anti-E cadherin monoclonal antibody DECMA-1. In contrast, DECMA-1 did not inhibit E cadherin-independent adhesion, such as the homotypic adhesion of Caco-2 cells in the absence of Ca(2+) or the heterotypic adhesion of Molt-3 T cells to Caco-2 monolayers. These results indicate the predominant involvement of E-cadherin mediated cell-cell adhesion in this assay. E-cadherin-derived peptides, which had been shown in our previous studies to inhibit E-cadherin-mediated cell-cell adhesion, significantly inhibited homotypic adhesion in a dose-dependent manner. These results, taken together, suggest that the present assay can be used for evaluation of peptide, protein, or antibody activity in modulating the E-cadherin mediated homophilic interactions in the context of whole live cells. PMID- 16371448 TI - Modeling glucocorticoid-mediated fetal lung maturation: II. Temporal patterns of gene expression in fetal rat lung. AB - Our previous report described the temporal steroid patterns during pharmacokinetic (PK) studies with dexamethasone (DEX) where doses of six 1 micromol/kg injections were given during gestational ages 18 to 20 days in rats. DEX PK was used in conjunction with the endogenous corticosterone profile to understand the regulation of fetal lung pharmacodynamics (PD). Expression of the glucocorticoid receptor (GR) and surfactant proteins A and B mRNA were chosen as lung maturational markers. GR seemed to be insensitive to the circulating glucocorticoids, indicating that unlike the adult situation, GR was not under negative feedback control of its ligand. Surfactant protein B exhibited approximately 400-fold induction in control fetal lung during the last days of gestation, and the inductive effect was even greater in the treatment group. Surfactant protein A displayed approximately 100-fold induction in control fetal lung during late gestation. However, the treatment group exhibited biphasic stimulatory and inhibitory effects for surfactant protein A. The inhibitory effect indicated that the chosen dosing scheme for DEX was not an optimal regimen. These data were used to determine by simulation the DEX regimen that would reproduce the temporal pattern of lung maturation observed in control animals. PK/PD modeling indicated that maintaining steroid exposure at approximately twice the equilibrium dissociation constant for the steroid/receptor interaction should produce optimal stimulation of both surfactant proteins. The simulations illustrate that administering smaller quantities of steroids over extended periods that produce sustained steroid exposure might be the optimal approach for designing dose-sparing antenatal corticosteroid therapy. PMID- 16371449 TI - Modeling glucocorticoid-mediated fetal lung maturation: I. Temporal patterns of corticosteroids in rat pregnancy. AB - Preterm birth produces neonatal respiratory distress syndrome, and dexamethasone (DEX) is administered maternally to induce fetal lung maturation in women at risk of preterm delivery. Antenatal DEX therapy is largely empirical, and administering multiple doses of DEX produces undesirable metabolic and developmental effects in the fetus. It is hypothesized that pharmacokinetic/pharmacodynamic (PK/PD) assessment of the maternal/fetal disposition and selected effects of corticosteroids will allow insight into optimal dosing methods. An optimal regimen was defined as a dosing schedule that would reproduce the endogenous prenatal steroid exposure and up-regulation of fetal lung maturational markers precociously. This report focuses on designing such a regimen from a PK standpoint in rats. The temporal profile of endogenous corticosterone in control rats was captured using a radioimmunoassay and showed that maternal and fetal corticosterone increased significantly during the last days of gestation. Six 1-mumol kg(-1) intramuscular DEX doses separated by 12-h intervals were administered maternally starting at gestational age 18, and PK was captured using a liquid chromatography-mass spectrometry assay. Unbound DEX exhibited a fetal to maternal concentration ratio of 0.55, had a free fraction of 0.2 in maternal and 0.4 in fetal plasma, and declined with a half-life of approximately 3 h. DEX PK and plasma protein binding were linear during the study, and DEX exposure caused severe adrenosuppression. These temporal steroid profiles in the fetal circulation will be used to drive the PD effects reported in a companion paper and an optimal steroid regimen will be proposed. PMID- 16371450 TI - 3H-1,2-dithiole-3-thione targets nuclear factor kappaB to block expression of inducible nitric-oxide synthase, prevents hypotension, and improves survival in endotoxemic rats. AB - Septicemia is a major cause of death associated with noncoronary intensive care. Systemic production of nitric oxide (NO) by inducible nitric-oxide synthase (iNOS) is a major cause of hypotension and poor organ perfusion seen in septic shock. Here, we show that pretreatment of F344 rats with the cancer chemoprotective agent 3H-1,2-dithiole-3-thione (D3T) blocks lipopolysaccharide (LPS)-mediated induction of hepatic iNOS and significantly reduces the associated serum levels of NO metabolites and enzyme markers of toxicity provoked by treatment with LPS. Immunohistochemical analysis shows that this protective effect is largely due to suppression of iNOS expression in hepatocytes. Importantly, pretreatment of animals with D3T blunts LPS-mediated hypotension and dramatically increases their survival. Inasmuch as iNOS expression can be regulated by nuclear factor (NF) kappaB, mechanistic studies show that D3T blocks NFkappaB nuclear translocation and DNA binding and that these effects are accompanied by changes in the levels of phospho-inhibitor of NFkappaB. In conclusion, this study identifies new drug classes and targets that may improve the prevention and treatment of septic shock, as well as chronic diseases associated with the NFkappaB and iNOS pathways. PMID- 16371451 TI - Comments on "Isobolographic analysis for combinations of a full and partial agonist: curved isoboles". PMID- 16371452 TI - Dynamic ensemble coding of saccades in the monkey superior colliculus. AB - The deeper layers of the midbrain superior colliculus (SC) contain a topographic motor map in which a localized population of cells is recruited for each saccade, but how the brain stem decodes the dynamic SC output is unclear. Here we analyze saccade-related responses in the monkey SC to test a new dynamic ensemble-coding model, which proposes that each spike from each saccade-related SC neuron adds a fixed, site-specific contribution to the intended eye movement command. As predicted by this simple theory, we found that the cumulative number of spikes in the cell bursts is tightly related to the displacement of the eye along the ideal straight trajectory, both for normal saccades and for strongly curved, blink perturbed saccades toward a single visual target. This dynamic relation depends systematically on the metrics of the saccade displacement vector, and can be fully predicted from a quantitative description of the cell's classical movement field. Furthermore, we show that a linear feedback model of the brain stem, which is driven by dynamic linear vector summation of measured SC firing patterns, produces realistic two-dimensional (2D) saccade trajectories and kinematics. We conclude that the SC may act as a nonlinear, vectorial saccade generator that programs an optimal straight eye-movement trajectory. PMID- 16371453 TI - Noradrenergic modulation of activity in a vocal control nucleus in vitro. AB - Norepinephrine (NE) can profoundly modulate sensory processing, but its effect on motor function is less well understood. Birdsong is a learned behavior involving sensory and motor processes that are influenced by NE. A potential site of NE action is the robust nucleus of the arcopallium (RA): RA receives noradrenergic inputs and has adrenergic receptors, and it is a sensorimotor area instrumental to song production. We hypothesized that NE modulates RA neurons, and as a first test, we examined the effect of NE on RA activity in vitro. We recorded spontaneous activity extracellularly from isolated RA neurons in brain slices made from adult male zebra finches. These neurons exhibited regular tonic activity with firing rates averaging 5.5 Hz. Bath application of NE rapidly and reversibly decreased firing for the majority of neurons, to the extent that spontaneous activity was often abolished. This was likely a direct effect on the cell recorded, because it occurred with blockade of fast excitatory and inhibitory synaptic transmission or of all synaptic transmission. The NE-induced suppression involved alpha2-adrenergic receptors: yohimbine, an antagonist, completely reversed the suppression, and clonidine, an agonist, partially mimicked it. Perforated patch recordings revealed that NE induced a conductance increase in RA neurons; however, this did not prevent cells from firing when stimulated by afferents in HVC. For some neurons, NE application resulted in an increase in signal-to-noise ratio for spikes evoked by HVC stimulation. Thus NE could strongly modulate the spontaneous activity of RA cells, potentially enhancing signals relayed through RA. PMID- 16371454 TI - Shift of intracellular chloride concentration in ganglion and amacrine cells of developing mouse retina. AB - GABA and glycine provide excitatory action during early development: they depolarize neurons and increase intracellular calcium concentration. As neurons mature, GABA and glycine become inhibitory. This switch from excitation to inhibition is thought to result from a shift of intracellular chloride concentration ([Cl-]i) from high to low, but in retina, measurements of [Cl-]i or chloride equilibrium potential (ECl) during development have not been made. Using the developing mouse retina, we systematically measured [Cl-]i in parallel with GABA's actions on calcium and chloride. In ganglion and amacrine cells, fura-2 imaging showed that before postnatal day (P) 6, exogenous GABA, acting via ionotropic GABA receptors, evoked calcium rise, which persisted in HCO3- -free buffer but was blocked with 0 extracellular calcium. After P6, GABA switched to inhibiting spontaneous calcium transients. Concomitant with this switch we observed the following: 6-methoxy-N-ethylquinolinium iodide (MEQ) chloride imaging showed that GABA caused an efflux of chloride before P6 and an influx afterward; gramicidin-perforated-patch recordings showed that the reversal potential for GABA decreased from -45 mV, near threshold for voltage-activated calcium channel, to -60 mV, near resting potential; MEQ imaging showed that [Cl ]i shifted steeply around P6 from 29 to 14 mM, corresponding to a decline of ECl from -39 to -58 mV. We also show that GABAergic amacrine cells became stratified by P4, potentially allowing GABA's excitatory action to shape circuit connectivity. Our results support the hypothesis that a shift from high [Cl-]i to low causes GABA to switch from excitatory to inhibitory. PMID- 16371455 TI - Roles of inhibition in complex auditory responses in the inferior colliculus: inhibited combination-sensitive neurons. AB - We studied the functional properties and underlying neural mechanisms associated with inhibitory combination-sensitive neurons in the mustached bat's inferior colliculus (IC). In these neurons, the excitatory response to best frequency tones was suppressed by lower frequency signals (usually in the range of 12-30 kHz) in a time-dependant manner. Of 143 inhibitory units, the majority (71%) were type I, in which low-frequency sounds evoked inhibition only. In the remainder, however, the low-frequency inhibitory signal also evoked excitation. Of these, excitation preceded the inhibition in type E/I units (16%), whereas in type I/E units (13%), excitation followed the inhibition. Type E/I and I/E units were distinct in the tuning and threshold sensitivity of low-frequency responses, whereas type I units overlapped the other types in these features. In 71 neurons, antagonists to receptors for glycine [strychnine (STRY)] or GABA [bicuculline (BIC)] were applied microiontophoretically. These antagonists failed to eliminate combination-sensitive inhibition in 92% (STRY), 93% (BIC), and 87% (BIC + STRY) of the type I units tested. However, inhibition was reduced in many neurons. Results were similar for type E/I and I/E inhibitory neurons. The results indicate that there are distinct populations of combination-sensitive inhibited neurons in the IC and that these populations are at least partly independent of glycine or GABAA receptors in the IC. We propose that these populations originate in different brain stem auditory nuclei, that they may be modified by interactions within the IC, and that they may perform different spectrotemporal analyses of vocal signals. PMID- 16371456 TI - Ca2+ influx, but not Ca2+ release from internal stores, is required for the PACAP induced increase in excitability in guinea pig intracardiac neurons. AB - Mechanisms modulating the pituitary adenylate cyclase activating polypeptide (PACAP)-induced increase in excitability have been studied using dissociated guinea pig intrinsic cardiac neurons and intact ganglion preparations. Measurements of intracellular calcium (Ca2+) with the fluorescent Ca2+ indicator dye fluo-3 indicated that neither PACAP nor vasoactive intestinal polypeptide (VIP) at either 100 nM or 1 microM produced a discernible elevation of intracellular Ca2+ in dissociated intracardiac neurons. For neurons in ganglion whole mount preparations kept in control bath solution, local application of PACAP significantly increased excitability, as indicated by the number of action potentials generated by long depolarizing current pulses. However, in a Ca2+ deficient solution in which external Ca2+ was replaced by Mg2+ or when cells were bathed in control solution containing 200 microM Cd2+, PACAP did not enhance action potential firing. In contrast, in a Ca2+ -deficient solution with Ca2+ replaced by strontium (Sr2+), PACAP increased excitability. PACAP increased excitability in cells treated with a combination of 20 microM ryanodine and 10 mM caffeine to interrupt release of Ca2+ from internal stores. Experiments using fluo-3 showed that ryanodine/caffeine pretreatment eliminated subsequent caffeine induced Ca2+ release from intracellular stores, whereas exposure to the Ca2+ deficient solution did not. In dissociated intracardiac neurons voltage clamped with the perforated patch recording technique, 100 nM PACAP decreased the voltage dependent barium current (IBa). These results show that, in the guinea pig intracardiac neurons, the PACAP-induced increase in excitability apparently requires Ca2+ influx through Cd2+ -sensitive calcium permeable channels other than voltage-dependent Ca2+ channels, but not Ca2+ release from internal stores. PMID- 16371457 TI - Feedback effects of horizontal cell membrane potential on cone calcium currents studied with simultaneous recordings. AB - Horizontal cell (HC) to cone feedback helps establish the center-surround arrangement of visual receptive fields. It has been shown that HC activity influences cone synaptic output by altering the amplitude and voltage dependence of the calcium current (ICa) in cones. In this study, we obtained voltage-clamp recordings simultaneously from cones and HCs to directly control the membrane potential of HCs and thereby measure the influence of HC membrane potential changes on ICa in adjacent cones. Directly hyperpolarizing voltage clamped HCs produced a negative activation shift and increased the amplitude of ICa in cones. Both of these effects were abolished by enhancing extracellular pH buffering capacity with HEPES. In contrast, addition of the gap junction blocker, carbenoxolone, did not significantly alter the shifts or amplitude changes in cone ICa produced by changes in HC membrane potential. These results support the hypothesis that changes in the HC membrane potential alter the voltage dependence and amplitude of cone ICa by altering extracellular pH levels at the synapse. PMID- 16371458 TI - Differences in the accuracy of human visuospatial memory after yaw and roll rotations. AB - Our ability to keep track of objects in the environment, even as we move, has been attributed to various cues including efference copies, vestibular signals, proprioception, and gravitational cues. However, the presence of some cues, such as gravity, may not be used to the same extent by different axes of motion (e.g., yaw vs. roll). We tested whether changes in gravitational cues can be used to improve visuospatial updating performance for yaw rotations as previously shown for roll. We found differences in updating for yaw and roll rotations in that yaw updating is not only associated with larger systematic errors but is also not facilitated by gravity in the same way as roll updating. PMID- 16371459 TI - Neuronal firing before and after burst discharges in the monkey basal ganglia is predictably patterned in the normal state and altered in parkinsonism. AB - It is known that burst discharges in basal ganglia neurons are more common in parkinsonism than under normal conditions, but changes in the structure of burst or peri-burst epochs have not been reported. In this study, the temporal structure of bursts and the timing of neuronal discharges that precede or follow them were examined in neuronal spike trains recorded in the subthalamic nucleus (STN) and the external and internal pallidal segment (GPe, GPi) in two awake Rhesus monkeys before and after they were rendered hemiparkinsonian by unilateral intracarotid infusion of the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). Bursts were detected by the "surprise" method. In the normal state, interspike intervals (ISIs) preceding or following bursts were frequently significantly longer than the average baseline ISI, and their duration was correlated with the burst length (i.e., the number of spikes/burst). Significant correlations were also found in all three structures between the burst length and the duration of interburst intervals. The incidence of burst discharges and the proportion of time spent in bursts increased in GPe, STN, and GPi after MPTP treatment. Burst lengths became more tightly related to preburst ISIs in the STN after MPTP treatment and to postburst ISI duration in all three structures. These results show that bursts in spontaneous GPe, STN, and GPi discharge are often preceded or followed by long ISIs, and that burst length, the length of pre- and postburst ISIs, and the length of interburst intervals are related to one another. Complex changes in these interactions may contribute to abnormal information processing in parkinsonism. PMID- 16371460 TI - V600E B-Raf requires the Hsp90 chaperone for stability and is degraded in response to Hsp90 inhibitors. AB - The Raf family includes three members, of which B-Raf is frequently mutated in melanoma and other tumors. We show that Raf-1 and A-Raf require Hsp90 for stability, whereas B-Raf does not. In contrast, mutated, activated B-Raf binds to an Hsp90-cdc37 complex, which is required for its stability and function. Exposure of melanoma cells and tumors to the Hsp90 inhibitor 17-allylamino-17 demethoxygeldanamycin results in the degradation of mutant B-Raf, inhibition of mitogen-activated protein kinase activation and cell proliferation, induction of apoptosis, and antitumor activity. These data suggest that activated mutated B Raf proteins are incompetent for folding in the absence of Hsp90, thus suggesting that the chaperone is required for the clonal evolution of melanomas and other tumors that depend on this mutation. Hsp90 inhibition represents a therapeutic strategy for the treatment of melanoma. PMID- 16371461 TI - Evidence for the direct involvement of {beta}TrCP in Gli3 protein processing. AB - Hedgehog-regulated processing of the transcription factor cubitus interruptus (Ci) in Drosophila depends on phosphorylation of the C-terminal region of Ci by cAMP-dependent protein kinase and subsequently by casein kinase 1 and glycogen synthase kinase 3. Ci processing also requires Slimb, an F-box protein of SCF (Skp1/Cullin/F-box proteins) complex, and the proteasome, but the interplay between phosphorylation and the activity of Slimb and the proteasome remains unclear. Here we show that processing of the Gli3 protein, a homolog of Ci, also depends on phosphorylation of a set of four cAMP-dependent protein kinase sites that primes subsequent phosphorylation of adjacent casein kinase 1 and glycogen synthase kinase 3. Our gain- and loss-of-function analyses in cultured cells further reveal that betaTrCP, the vertebrate homolog of Slimb, is required for Gli3 processing, and we demonstrate that betaTrCP can bind phosphorylated Gli3 both in vitro and in vivo. We also find that the Gli3 protein is polyubiquitinated in the cell and that its processing depends on proteasome activity. Our findings provide evidence for a direct link between phosphorylation of Gli3/Ci proteins and betaTrCP/Slimb action, thus supporting the hypothesis that the processing of Gli3/Ci is affected by the proteasome. PMID- 16371462 TI - The questionable contribution of the Neolithic and the Bronze Age to European craniofacial form. AB - Many human craniofacial dimensions are largely of neutral adaptive significance, and an analysis of their variation can serve as an indication of the extent to which any given population is genetically related to or differs from any other. When 24 craniofacial measurements of a series of human populations are used to generate neighbor-joining dendrograms, it is no surprise that all modern European groups, ranging all of the way from Scandinavia to eastern Europe and throughout the Mediterranean to the Middle East, show that they are closely related to each other. The surprise is that the Neolithic peoples of Europe and their Bronze Age successors are not closely related to the modern inhabitants, although the prehistoric/modern ties are somewhat more apparent in southern Europe. It is a further surprise that the Epipalaeolithic Natufian of Israel from whom the Neolithic realm was assumed to arise has a clear link to Sub-Saharan Africa. Basques and Canary Islanders are clearly associated with modern Europeans. When canonical variates are plotted, neither sample ties in with Cro-Magnon as was once suggested. The data treated here support the idea that the Neolithic moved out of the Near East into the circum-Mediterranean areas and Europe by a process of demic diffusion but that subsequently the in situ residents of those areas, derived from the Late Pleistocene inhabitants, absorbed both the agricultural life way and the people who had brought it. PMID- 16371463 TI - Regulation of dendritic cell maturation and function by Bruton's tyrosine kinase via IL-10 and Stat3. AB - Btk plays crucial roles in the differentiation and activation of B and myeloid cells. Despite drastic reductions of other Ig isotypes, paradoxically high IgE responses have been known in btk mutant mice. Here we show that btk(-/-) dendritic cells exhibit a more mature phenotype and a stronger in vitro and in vivo T cell-stimulatory ability than wild-type cells. Increased IgE responses were induced by adoptive transfer of btk(-/-) dendritic cells into mice. Consistent with the stronger T cell-stimulatory ability of btk(-/-) dendritic cells, btk(-/-) mice exhibited enhanced inflammation in Th2-driven asthma and Th1 driven contact sensitivity experiments. These negative regulatory functions of Btk in dendritic cells appear to be mediated mainly through autocrine secretion of IL-10 and subsequent activation of Stat3. PMID- 16371464 TI - Gialpha and Gbeta subunits both define selectivity of G protein activation by alpha2-adrenergic receptors. AB - Previous studies of the specificity of receptor interactions with G protein subunits in living cells have relied on measurements of second messengers or other downstream responses. We have examined the selectivity of interactions between alpha2-adrenergic receptors (alpha2R) and various combinations of Gialpha and Gbeta subunit isoforms by measuring changes in FRET between Gialpha-yellow fluorescent protein and cyan fluorescent protein-Gbeta chimeras in HeLa cells. All combinations of Gialpha1, -2, or -3 with Gbeta1, -2, or -4 were activated to some degree by endogenous alpha2Rs as judged by agonist-dependent decreases in FRET. The degree of G protein activation is determined by the combination of Gialpha and Gbeta subunits rather than by the identity of an individual subunit. RT-PCR analysis and small interfering RNA knockdown of alpha2R subtypes, followed by quantification of radiolabeled antagonist binding, demonstrated that HeLa cells express alpha2a- and alpha2b-adrenergic receptor isoforms in a 2:1 ratio. Increasing receptor number by overexpression of the alpha2aR subtype minimized the differences among coupling preferences for Gialpha and Gbeta isoforms. The molecular properties of each Gialpha, Gbeta, and alpha2-adrenergic receptor subtype influence signaling efficiency for the alpha2-adrenergic receptor mediated signaling pathway. PMID- 16371465 TI - CYP27B1 null mice with LacZreporter gene display no 25-hydroxyvitamin D3-1alpha hydroxylase promoter activity in the skin. AB - The hormonally active form of vitamin D(3),1alpha,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], is synthesized in the kidney through a tightly regulated reaction catalyzed by 25-hydroxyvitamin D(3)-1alpha-hydroxylase (1alpha hydroxylase), the product of the CYP27B1 gene. Through gene targeting in embryonic stem cells, we engineered a mouse strain in which the coding region of the 1alpha-hydroxylase gene is replaced by the genes for beta-galactosidase (lacZ) and neomycin resistance. Null mice produced no detectable 1alpha hydroxylase transcript. The mice grew normally when maintained on a balanced diet containing 1,25(OH)(2)D(3) but rapidly developed rickets when phosphorus and 1,25(OH)(2)D(3) were restricted. Rickets was curable through administration of 1,25(OH)(2)D(3) but not its biological precursor, 25-hydroxyvitamin D(3). Upon administration of a diet low in calcium and devoid of any form of vitamin D(3), beta-galactosidase activity was detected in the kidneys of the -/- and +/- mice and in placentas harvested from -/- females bred with -/- males. No beta galactosidase activity was detected in skin sections or in primary keratinocyte cultures from -/- animals. Our results demonstrate we have generated 1alpha hydroxylase null mice that display phenotypes characteristic of vitamin D dependency rickets type I. From the histochemical analysis of reporter gene expression in these mice, we conclude that acute 1,25(OH)(2)D(3) deficiency in otherwise healthy animals does not stimulate local production of 1,25(OH)(2)D(3) in the skin. These findings stand in contrast to previously published reports of 1,25(OH)(2)D(3) production in keratinocytes. PMID- 16371466 TI - Global landscape of recent inferred Darwinian selection for Homo sapiens. AB - By using the 1.6 million single-nucleotide polymorphism (SNP) genotype data set from Perlegen Sciences [Hinds, D. A., Stuve, L. L., Nilsen, G. B., Halperin, E., Eskin, E., Ballinger, D. G., Frazer, K. A. & Cox, D. R. (2005) Science 307, 1072 1079], a probabilistic search for the landscape exhibited by positive Darwinian selection was conducted. By sorting each high-frequency allele by homozygosity, we search for the expected decay of adjacent SNP linkage disequilibrium (LD) at recently selected alleles, eliminating the need for inferring haplotype. We designate this approach the LD decay (LDD) test. By these criteria, 1.6% of Perlegen SNPs were found to exhibit the genetic architecture of selection. These results were confirmed on an independently generated data set of 1.0 million SNP genotypes (International Human Haplotype Map Phase I freeze). Simulation studies indicate that the LDD test, at the megabase scale used, effectively distinguishes selection from other causes of extensive LD, such as inversions, population bottlenecks, and admixture. The approximately 1,800 genes identified by the LDD test were clustered according to Gene Ontology (GO) categories. Based on overrepresentation analysis, several predominant biological themes are common in these selected alleles, including host-pathogen interactions, reproduction, DNA metabolism/cell cycle, protein metabolism, and neuronal function. PMID- 16371467 TI - Molecular cloning, expression, and structural prediction of deoxyhypusine hydroxylase: a HEAT-repeat-containing metalloenzyme. AB - The eukaryotic initiation factor 5A (eIF5A), a factor essential for eukaryotic cell proliferation, is the only cellular protein containing the polyamine-derived amino acid hypusine [N(epsilon)-(4-amino-2-hydroxybutyl)lysine]. Hypusine is formed in a posttranslational modification that involves two sequential enzymatic steps catalyzed by deoxyhypusine synthase and deoxyhypusine hydroxylase (DOHH). By screening a Saccharomyces cerevisiae GST-ORF library for expression of DOHH activity, we have cloned YJR070C as the gene encoding DOHH and identified the human homolog DOHH gene, HLRC1. Purified recombinant yeast and human DOHH enzymes effectively catalyzed hydroxylation of the deoxyhypusine residue in the eIF5A intermediate. Overexpression of human DOHH along with eIF5A precursor and deoxyhypusine synthase was required for overproduction of mature, hypusine containing eIF5A in 293T and other mammalian cells. The Saccharomyces cerevisiae strain with deletion of YJR070C contained only deoxyhypusine but no hypusine, indicating that YJR070C was the single DOHH gene in this organism. One highly conserved DOHH homolog gene is found in a variety of eukaryotes from yeast to human. Sequence and structural analyses reveal that DOHH belongs to a family of HEAT-repeat-containing proteins, consisting of eight tandem repeats of an alpha helical pair (HEAT motif) organized in a symmetrical dyad. The predicted structure is unrelated to the double-stranded beta-helix type structures of the Fe(II)- and 2-oxoacid-dependent dioxygenases, such as collagen prolyl or lysyl hydroxylases. However, metal coordination sites composed of four strictly conserved histidine-glutamate sequences were identified, suggesting that DOHH enzymes have convergently evolved an iron-dependent hydroxylation mechanism. PMID- 16371468 TI - Determination of an ensemble of structures representing the intermediate state of the bacterial immunity protein Im7. AB - We present a detailed structural characterization of the intermediate state populated during the folding and unfolding of the bacterial immunity protein Im7. We achieve this result by incorporating a variety of experimental data available for this species in molecular dynamics simulations. First, we define the structure of the exchange-competent intermediate state of Im7 by using equilibrium hydrogen-exchange protection factors. Second, we use this ensemble to predict Phi-values and compare the results with the experimentally determined Phi values of the kinetic refolding intermediate. Third, we predict chemical-shift measurements and compare them with the measured chemical shifts of a mutational variant of Im7 for which the kinetic folding intermediate is the most stable state populated at equilibrium. Remarkably, we found that the properties of the latter two species are predicted with high accuracy from the exchange-competent intermediate that we determined, suggesting that these three states are characterized by a similar architecture in which helices I, II, and IV are aligned in a native-like, but reorganized, manner. Furthermore, the structural ensemble that we obtained enabled us to rationalize the results of tryptophan fluorescence experiments in the WT protein and a series of mutational variants. The results show that the integration of diverse sets of experimental data at relatively low structural resolution is a powerful approach that can provide insights into the structural organization of this conformationally heterogeneous three-helix intermediate with unprecedented detail and highlight the importance of both native and non-native interactions in stabilizing its structure. PMID- 16371469 TI - Inhibitory control by an integral feedback signal in prefrontal cortex: a model of discrimination between sequential stimuli. AB - The prefrontal cortex (PFC) is known to be critical for inhibitory control of behavior, but the underlying mechanisms are unclear. Here, we propose that inhibitory control can be instantiated by an integral signal derived from working memory, another key function of the PFC. Specifically, we assume that an integrator converts excitatory input into a graded mnemonic activity that provides an inhibitory signal (integral feedback control) to upstream afferent neurons. We demonstrate this scenario in a neuronal-network model for a temporal discrimination task. The task requires the working memory of the vibrational frequency (f1) of an initial stimulus (stimulus 1), followed by comparison of the frequency (f2) of a second stimulus (stimulus 2) with the stored f1 and a binary decision (f2 > f1 or f2 < f1). The integral feedback signal generated by stimulus 1 gates the later inputs based on the amplitude difference (f2 - f1). The feedback control signal enables a subset of neurons to reverse their tuning to f1 between stimulus 1 and stimulus 2, when they become tuned to the difference, f2 - f1. These neurons maintain a lower firing rate during the delay compared with their peak rate during stimulus 1. A second subset of neurons, tuned to f1 during the delay, reaches a rate during stimulus 2 that depends on the maximum of f1 and f2. Our work suggests a circuit mechanism for discrimination across time and predicts neuronal behavior that can be tested experimentally. PMID- 16371470 TI - A gradient of auxin and auxin-dependent transcription precedes tropic growth responses. AB - Plants, although sessile, can reorient growth axes in response to changing environmental conditions. Phototropism and gravitropism represent adaptive growth responses induced by changes in light direction and growth axis orientation relative to gravitational direction, respectively. The nearly 80-year-old Cholodny-Went theory [Went, F. W. & Thimann, K. V. (1937) Phytohormones (Macmillan, New York)] predicts that formation of a gradient of the plant morphogen auxin is central to the establishment of tropic curvature. Loss of tropic responses in seedling stems of Arabidopsis thaliana mutants lacking the auxin-regulated transcriptional activator NPH4/ARF7 has further suggested that a gradient of gene expression represents an essential output from the auxin gradient. Yet the molecular identities of such output components, which are likely to encode proteins directly involved in growth control, have remained elusive. Here we report the discovery of a suite of tropic stimulus-induced genes in Brassica oleracea that are responsive to an auxin gradient and exhibit morphologically graded expression concomitant with, or before, observable curvature responses. These results provide compelling molecular support for the Cholodny-Went theory and suggest that morphologically graded transcription represents an important mechanism for interpreting tropically stimulated gradients of auxin. Intriguingly, two of the tropic stimulus-induced genes, EXPA1 and EXPA8, encode enzymes involved in cell wall extension, a response prerequisite for differential growth leading to curvatures, and are up-regulated before curvature in the flank that will elongate. This observation suggests that morphologically graded transcription likely leads to the graded expression of proteins whose activities can directly regulate the establishment and modulation of tropic curvatures. PMID- 16371471 TI - The Bacillus subtilis spore coat provides "eat resistance" during phagocytic predation by the protozoan Tetrahymena thermophila. AB - Bacillus spores are highly resistant to many environmental stresses, owing in part to the presence of multiple "extracellular" layers. Although the role of some of these extracellular layers in resistance to particular stresses is known, the function of one of the outermost layers, the spore coat, is not completely understood. This study sought to determine whether the spore coat plays a role in resistance to predation by the ciliated protozoan Tetrahymena, which uses phagocytosis to ingest and degrade other microorganisms. Wild-type dormant spores of Bacillus subtilis were efficiently ingested by the protozoan Tetrahymena thermophila but were neither digested nor killed. However, spores with various coat defects were killed and digested, leaving only an outer shell termed a rind, and supporting the growth of Tetrahymena. A similar rind was generated when coat defective spores were treated with lysozyme alone. The sensitivity of spores with different coat defects to predation by T. thermophila paralleled the spores' sensitivities to lysozyme. Spore killing by T. thermophila was by means of lytic enzymes within the protozoal phagosome, not by initial spore germination followed by killing. These findings suggest that a major function of the coat of spores of Bacillus species is to protect spores against predation. We also found that indigestible rinds were generated even from spores in which cross-linking of coat proteins was greatly reduced, implying the existence of a coat structure that is highly resistant to degradative enzymes. PMID- 16371472 TI - Two independent mechanical events in the interaction cycle of skeletal muscle myosin with actin. AB - During skeletal muscle contraction, regular arrays of actin and myosin filaments slide past each other driven by the cyclic ATP-dependent interaction of the motor protein myosin II (the cross-bridge) with actin. The rate of the cross-bridge cycle and its load-dependence, defining shortening velocity and energy consumption at the molecular level, vary widely among different isoforms of myosin II. However, the underlying mechanisms remain poorly understood. We have addressed this question by applying a single-molecule approach to rapidly ( approximately 300 mus) and precisely ( approximately 0.1 nm) detect acto-myosin interactions of two myosin isoforms having large differences in shortening velocity. We show that skeletal myosin propels actin filaments, performing its conformational change (working stroke) in two steps. The first step ( approximately 3.4-5.2 nm) occurs immediately after myosin binding and is followed by a smaller step ( approximately 1.0-1.3 nm), which occurs much faster in the fast myosin isoform than in the slow one, independently of ATP concentration. On the other hand, the rate of the second phase of the working stroke, from development of the latter step to dissociation of the acto-myosin complex, is very similar in the two isoforms and depends linearly on ATP concentration. The finding of a second mechanical event in the working stroke of skeletal muscle myosin provides the molecular basis for a simple model of actomyosin interaction. This model can account for the variation, in different fiber types, of the rate of the cross-bridge cycle and provides a common scheme for the chemo-mechanical transduction within the myosin family. PMID- 16371473 TI - Toll-like receptor (TLR) polymorphisms in African children: Common TLR-4 variants predispose to severe malaria. AB - Genetic host factors play a substantial role in susceptibility to and severity of malaria, which continues to cause at least one million deaths per year. Recently, members of the toll-like receptor (TLR) family have been shown to be involved in recognition of the etiologic organism Plasmodium falciparum: The glycosylphosphatidylinositol anchor induces signaling in host cells via TLR-2 and -4, whereas hemozoin-induced immune activation involves TLR-9. Binding of microbial ligands to the respective TLRs triggers the release of proinflammatory cytokines via the TLR/IL-1 receptor (TIR) domain and may contribute to the host response in malaria, including cytokine induction and fever. In a case-control study among 870 Ghanaian children, we examined the influence of TLR-2, -4, and -9 polymorphisms in susceptibility to severe malaria. TLR-2 variants common in Caucasians and Asians were completely absent. However, we found a rare previously undescribed mutation (Leu658Pro), which impairs signaling via TLR-2. We failed to detect any polymorphisms within the TLR-9 Toll/IL-1 receptor domain. Two frequent TLR-9 promoter polymorphisms did not show a clear association with malaria severity. In contrast, the TLR-4-Asp299Gly variant occurred at a high rate of 17.6% in healthy controls and was even more frequent in severe malaria patients (24.1%, P < 0.05). Likewise, TLR-4-Thr399Ile was seen in 2.4% of healthy children and in 6.2% of patients (P = 0.02). TLR-4-Asp299Gly and TLR-4-Thr399Ile conferred 1.5- and 2.6-fold increased risks of severe malaria, respectively. These findings suggest TLR4-mediated responses to malaria in vivo and TLR-4 polymorphisms to be associated with disease manifestation. PMID- 16371474 TI - Neural activity in speech-sensitive auditory cortex during silence. AB - That auditory hallucinations are voices heard in the absence of external stimuli implies the existence of endogenous neural activity within the auditory cortex responsible for their perception. Further, auditory hallucinations occur across a range of healthy and disease states that include reduced arousal, hypnosis, drug intoxication, delirium, and psychosis. This suggests that, even in health, the auditory cortex has a propensity to spontaneously "activate" during silence. Here we report the findings of a functional MRI study, designed to examine baseline activity in speech-sensitive auditory regions. During silence, we show that functionally defined speech-sensitive auditory cortex is characterized by intermittent episodes of significantly increased activity in a large proportion (in some cases >30%) of its volume. Bilateral increases in activity are associated with foci of spontaneous activation in the left primary and association auditory cortices and anterior cingulate cortex. We suggest that, within auditory regions, endogenous activity is modulated by anterior cingulate cortex, resulting in spontaneous activation during silence. Hence, an aspect of the brain's "default mode" resembles a (preprepared) substrate for the development of auditory hallucinations. These observations may help explain why such hallucinations are ubiquitous. PMID- 16371475 TI - Intraprotein transfer of the quinone analogue inhibitor 2,5-dibromo-3-methyl-6 isopropyl-p-benzoquinone in the cytochrome b6f complex. AB - Details are presented of the structural analysis of the cytochrome b(6)f complex from the thermophilic cyanobacterium, Mastigocladus laminosus, in the presence of the electrochemically positive (p)-side quinone analogue inhibitor, 2,5-dibromo-3 methyl-6-isopropylbenzoquinone (DBMIB). One DBMIB binding site was found. This site is peripheral to the quinone binding space defined by the binding sites of other p-side inhibitors previously resolved in cytochrome bc(1)/b(6)f complexes. This high-affinity site resides in a p-side interfacial niche bounded by cytochrome f, subunit IV, and cytochrome b(6), is close (8 A) to the p-side heme b, but distant (19 A) from the [2Fe-2S] cluster. No significant electron density associated with the DBMIB was found elsewhere in the structure. However, the site at which DBMIB can inhibit light-induced redox turnover is within a few A of the [2Fe-2S] cluster, as shown by the absence of inhibition in mutants of Synechococcus sp. PCC 7002 at iron sulfur protein-Leu-111 near the cluster. The ability of a minimum amount of initially oxidized DBMIB to inhibit turnover of WT complex after a second light flash implies that there is a light-activated movement of DBMIB from the distal peripheral site to an inhibitory site proximal to the [2Fe-2S] cluster. Together with the necessary passage of quinone/quinol through the small Q(p) portal in the complex, it is seen that transmembrane traffic of quinone-like molecules through the core of cytochrome bc complexes can be labyrinthine. PMID- 16371476 TI - PDSM, a motif for phosphorylation-dependent SUMO modification. AB - SUMO (small ubiquitin-like modifier) modification regulates many cellular processes, including transcription. Although sumoylation often occurs on specific lysines within the consensus tetrapeptide PsiKxE, other modifications, such as phosphorylation, may regulate the sumoylation of a substrate. We have discovered PDSM (phosphorylation-dependent sumoylation motif), composed of a SUMO consensus site and an adjacent proline-directed phosphorylation site (PsiKxExxSP). The highly conserved motif regulates phosphorylation-dependent sumoylation of multiple substrates, such as heat-shock factors (HSFs), GATA-1, and myocyte enhancer factor 2. In fact, the majority of the PDSM-containing proteins are transcriptional regulators. Within the HSF family, PDSM is conserved between two functionally distinct members, HSF1 and HSF4b, whose transactivation capacities are repressed through the phosphorylation-dependent sumoylation. As the first recurrent sumoylation determinant beyond the consensus tetrapeptide, the PDSM provides a valuable tool in predicting new SUMO substrates. PMID- 16371477 TI - Bisphosphonate-mediated gene vector delivery from the metal surfaces of stents. AB - The clinical use of metallic expandable intravascular stents has resulted in improved therapeutic outcomes for coronary artery disease. However, arterial reobstruction after stenting, in-stent restenosis, remains an important problem. Gene therapy to treat in-stent restenosis by using gene vector delivery from the metallic stent surfaces has never been demonstrated. The present studies investigated the hypothesis that metal-bisphosphonate binding can enable site specific gene vector delivery from metal surfaces. Polyallylamine bisphosphonate (PAA-BP) was synthesized by using Michael addition methodology. Exposure to aqueous solutions of PAA-BP resulted in the formation of a monomolecular bisphosphonate layer on metal alloy surfaces (steel, nitinol, and cobalt chromium), as demonstrated by x-ray photoelectron spectroscopy. Surface-bound PAA BP enabled adenoviral (Ad) tethering due to covalent thiol-binding of either anti Ad antibody or a recombinant Ad-receptor protein, D1. In arterial smooth muscle cell cultures, alloy samples configured with surface-tethered Ad were demonstrated to achieve site-specific transduction with a reporter gene, (GFP). Rat carotid stent angioplasties using metal stents exposed to aqueous PAA-BP and derivatized with anti-knob antibody or D1 resulted in extensive localized Ad-GFP expression in the arterial wall. In a separate study with a model therapeutic vector, Ad-inducible nitric oxide synthase (iNOS) attached to the bisphosphonate treated metal stent surface via D1, significant inhibition of restenosis was demonstrated (neointimal/media ratio 1.68 +/- 0.27 and 3.4 +/- 0.35; Ad-iNOS vs. control, P < 0.01). It is concluded that effective gene vector delivery from metallic stent surfaces can be achieved by using this approach. PMID- 16371478 TI - Does complete deficiency of muscle alpha actinin 3 alter functional capacity in elderly women? A preliminary report. AB - The sarcomeric protein alpha actinin 3 is localised to the Z line of fast fibres, which are responsible for generating forceful muscle contractions at high velocity. However, a substantial proportion of healthy humans are totally deficient in this protein as they are homozygous for a premature stop codon polymorphism (R577X) in the ACTN3 gene. The purpose of this preliminary study was to assess if the presence or absence of alpha actinin 3 influences the deleterious effects of ageing on muscle output and functional capacity. PMID- 16371479 TI - Concussion management by primary care providers. AB - OBJECTIVE: To assess current concussion management practices of primary care providers. METHODS: An 11 item questionnaire was mailed to primary care providers in the state of Maine, with serial mailings to non-respondents. RESULTS: Over 50% of the questionnaires were completed, with nearly 70% of primary care providers indicating that they routinely use published guidelines as a tool in managing patients with concussion. Nearly two thirds of providers were aware that neuropsychological tests could be used, but only 16% had access to such tests within a week of injury. CONCLUSIONS: Primary care providers are using published concussion management guidelines with high frequency, but many are unable to access neuropsychological testing when it is required. PMID- 16371480 TI - The end of the beginning. PMID- 16371481 TI - Building bone mass through exercise: could less be more? PMID- 16371482 TI - Benefits of exercise therapy for chronic diseases. PMID- 16371483 TI - Gene therapy in sport. PMID- 16371484 TI - Research based recommendations on management of sport related concussion: summary of the National Athletic Trainers' Association position statement. PMID- 16371485 TI - Effect of oral contraceptives and hormone replacement therapy on bone mineral density in premenopausal and perimenopausal women: a systematic review. AB - Seventy five articles on the effect of oral contraceptives and other hormone replacement on bone density in premenopausal and perimenopausal women were reviewed. The evidence was appraised using the Oxford Centre for Evidence-Based Medicine levels of evidence. There is good evidence for a positive effect of oral contraceptives on bone density in perimenopausal women, and fair evidence for a positive effect in "hypothalamic" oligo/amenorrhoeic premenopausal women. There is limited evidence for a positive effect in healthy and anorexic premenopausal women. In hypothalamic oligo/amenorrhoeic women, baseline bone density has been shown to be significantly lower than that in healthy controls, therefore the decision to treat is clinically more important. The ideal formulation(s) and duration of treatment remain to be determined by further longitudinal and prospective randomised controlled trials in larger subject populations. PMID- 16371486 TI - Graded associations between cardiorespiratory fitness, fatness, and blood pressure in children and adolescents. AB - OBJECTIVE: To measure the graded relation between cardiorespiratory fitness and sum of skinfolds, waist circumference, and blood pressure in children and adolescents participating in the European youth heart study. METHODS: The participants were 4072 children and adolescents (aged 9 and 15) from Denmark, Portugal, Estonia, and Norway. Cardiorespiratory fitness was indirectly determined using a maximal ergometer cycle test. The sum of four skinfolds, waist circumference, and blood pressure were assessed with a standardised protocol. Linear regression analysis was used to test the graded relation between cardiorespiratory fitness and the dependent variables adjusted for pubertal stage, sex, and country. RESULTS: A significant curvilinear graded relation was found between cardiorespiratory fitness and waist circumference and sum of skinfolds (partial r2 for cardiorespiratory fitness was 0.09-0.26 for the different sexes and age groups). Systolic and diastolic blood pressure also showed a curvilinear relation with cardiorespiratory fitness, and fitness explained 2% of the variance in systolic blood pressure. The difference in systolic blood pressure between the least and most fit was 6 mm Hg. CONCLUSION: A curvilinear graded relation was found between cardiorespiratory fitness and waist circumference, sum of skinfolds, and systolic blood pressure. The greatest difference in these health variables was observed between low and moderate fitness levels. PMID- 16371487 TI - Patellar taping does not change the amplitude of electromyographic activity of the vasti in a stair stepping task. AB - OBJECTIVES: To investigate the effect of patellar taping on the amplitude of electromyographic activity (EMG) of vasti activation in subjects with and without patellofemoral pain (PFP). METHODS: Ten participants with PFP and 12 asymptomatic controls were recruited to the study. The study was designed as a randomised crossover trial. Participants completed a stair stepping task. Three experimental conditions were assessed: no tape, therapeutic medially directed tape, and placebo vertically directed tape. The main outcome measure was the EMG amplitude of the vastus medialis obliquus and vastus lateralis during the concentric phase of stair stepping. RESULTS: The application of medially directed therapeutic tape significantly decreased pain in subjects with PFP. However, application of tape over the patella (therapeutic or placebo) did not alter the amplitude of vasti EMG when either the PFP or control participants completed the concentric stair stepping task. CONCLUSION: The results of this study indicate that the positive clinical effects of medially directed therapeutic tape are not due to changes in EMG amplitude of the vasti muscle. Thus other effects such as changes in timing of contraction of the vasti are more likely candidates for the mechanism of efficacy. PMID- 16371488 TI - Angiogenic effect of intramuscular administration of basic and acidic fibroblast growth factor on skeletal muscles and influence of exercise on muscle angiogenesis. AB - BACKGROUND: Angiogenic factors which control the angiogenic process represent a promising strategy for restoration of blood flow, but require further evaluation before clinical use. Exercise has also been reported to induce neovascularisation in muscles. OBJECTIVES: To evaluate the angiogenic effects of basic fibroblast growth factor (b-FGF) and acidic fibroblast growth factor (a-FGF) on rat gastrocnemius muscle, when administered intramuscularly, and to compare them with those obtained by daily exercise. METHODS: Forty nine rats were allotted to the following groups: A, controls; B, exercise by swimming; C1 and C2, intramuscular injection of b-FGF and a-FGF respectively; D1 and D2, b-FGF and a-FGF injection in combination with exercise. The antibody mouse anti-rat CD31 was used to evaluate the numbers of blood vessels present in histological preparations of gastrocnemius muscle. RESULTS: Significant increases in the numbers of blood vessels of the right gastrocnemius muscles in groups C1 and D1 were observed compared with controls (p<0.05). There was only a slight increase in angiogenesis in the left gastrocnemius muscle of groups C1 and D1 compared with controls (p>0.05), and there was a decrease in angiogenesis in the gastrocnemius muscle of the swimming group compared with controls. CONCLUSION: The intramuscular administration of b-FGF, but not a-FGF, induced significant local angiogenesis in gastrocnemius muscle at the site of injection. PMID- 16371489 TI - Type of acute hamstring strain affects flexibility, strength, and time to return to pre-injury level. AB - OBJECTIVES: To investigate possible links between aetiology of acute, first time hamstring strains in sprinters and dancers and recovery of flexibility, strength, and function as well as time to return to pre-injury level. [figure: see text]. METHODS: Eighteen elite sprinters and 15 professional dancers with a clinically diagnosed hamstring strain were included. They were clinically examined and tested two, 10, 21, and 42 days after the acute injury. Range of motion in hip flexion and isometric strength in knee flexion were measured. Self estimated and actual time to return to pre-injury level were recorded. Hamstring reinjuries were recorded during a two year follow up period. RESULTS: All the sprinters sustained their injuries during high speed sprinting, whereas all the dancers were injured while performing slow stretching type exercises. The initial loss of flexibility and strength was greater in sprinters than in dancers (p<0.05). At 42 days after injury, both groups could perform more than 90% of the test values of the uninjured leg. However, the actual times to return to pre-injury level of performance were significantly longer (median 16 weeks (range 6-50) for the sprinters and 50 weeks (range 30-76) for the dancers). Three reinjuries were noted, all in sprinters. CONCLUSION: There appears to be a link between the aetiologies of the two types of acute hamstring strain in sprinters and dancers and the time to return to pre-injury level. Initially, sprinters have more severe functional deficits but recover more quickly. PMID- 16371490 TI - Hip passive range of motion and frequency of radiographic hip osteoarthritis in former elite handball players. AB - OBJECTIVE: To establish the relation between handball playing, passive hip range of motion (ROM), and the development of radiological hip osteoarthritis (OA) in former elite handball players. Two related issues are addressed: (a) the relation between long term elite handball playing and the incidence of hip OA; (b) the relations between hip ROM, OA, and pain. METHODS: Data on 20 former elite handball players and 39 control subjects were collected. A questionnaire yielded personal details, loading patterns during physical activity, and previous lower limb joint injury. Bilateral radiographs were analysed to diagnose and classify hip OA. Passive hip ROM was measured bilaterally with a goniometer. RESULTS: A close relation was found between long term elite handball practice and the incidence of hip OA: 60% of the handball players were diagnosed with OA in at least one of the hip joints compared with 13% of the control subjects. Passive ROM measured in the handball players was significantly lower for hip flexion and medial rotation and higher for abduction, extension, and lateral rotation than the control values. The handball players with OA reported less pain in the hip joints during daily activities than the control subjects with OA. CONCLUSION: The risk of developing premature hip OA seems high for retired handball players and significantly greater than for the general population. Pain and discomfort represent two difficult diagnostic challenges to the sports physician, as the repetitive nature of movements that are specific to handball can lead to alterations that are rarely seen in the general population. PMID- 16371491 TI - Effects of long term Tai Chi practice and jogging exercise on muscle strength and endurance in older people. AB - OBJECTIVES: To investigate the influence of regular Tai Chi (TC) practice and jogging on muscle strength and endurance in the lower extremities of older people. METHODS: Twenty one long term older TC practitioners were compared with 18 regular older joggers and 22 sedentary counterparts. Maximum concentric strength of knee flexors and extensors was tested at angular velocities of 30 degrees/s and 120 degrees/s. Ankle dorsiflexors and plantar flexors were tested at 30 degrees/s and the dynamic endurance of the knee flexors and extensors was assessed at a speed of 180 degrees/s. RESULTS: The differences in the muscle strength of the knee joint amongst the three experimental groups were significant at the higher velocity. The strengths of knee extensors and flexors in the control group were significantly lower than those in the jogging group and marginally lower than those in the TC group. For the ankle joint, the subjects in both the TC and jogging groups generated more torque in their ankle dorsiflexors. In addition, the muscle endurance of knee extensors was more pronounced in TC practitioners than in controls. CONCLUSION: Regular older TC practitioners and joggers showed better scores than the sedentary controls on most muscle strength and endurance measures. However, the magnitude of the exercise effects on muscles might depend on the characteristics of different types of exercise. PMID- 16371492 TI - Acute weight loss followed by an aggressive nutritional recovery strategy has little impact on on-water rowing performance. AB - OBJECTIVES: To assess the influence of moderate, acute weight loss on on-water rowing performance when aggressive nutritional recovery strategies were used in the two hours between weigh in and racing. METHODS: Competitive rowers (n = 17) undertook three on-water 1800 m time trials under cool conditions (mean (SD) temperature 8.4 (2.0) degrees C), each separated by 48 hours. No weight limit was imposed for the first time trial--that is, unrestricted body mass (UNR1). However, one of the remaining two trials followed a 4% loss in body mass in the previous 24 hours (WT(-4%)). No weight limit was imposed for the other trial (UNR2). Aggressive nutritional recovery strategies (WT(-4%), 2.3 g/kg carbohydrate, 34 mg/kg Na+, and 28.4 ml/kg fluid; UNR, ad libitum) were used in the first 90 minutes of the two hours between weigh in and performance trials. RESULTS: WT(-4%) had only a small and statistically non-significant effect on the on-water time trial performance (mean 1.0 second, 95% confidence interval (CI) 0.9 to 2.8; p = 0.29) compared with UNR. This was despite a significant decrease in plasma volume at the time of weigh in for WT(-4%) compared with UNR (-9.2%, 95% CI -12.8% to -5.6%; p<0.001). CONCLUSIONS: Acute weight loss of up to 4% over 24 hours, when combined with aggressive nutritional recovery strategies, can be undertaken with minimal impact on on-water rowing performance, at least in cool conditions. PMID- 16371493 TI - Effects of brief yoga exercises and motivational preparatory interventions in distance runners: results of a controlled trial. AB - OBJECTIVE: To examine the efficacy of two preparatory interventions on one mile run performance in 90 high school long distance runners. METHOD: After participants had completed a one mile baseline run, they were randomly assigned to participate in either one of two interventions (brief yoga exercises, motivational shouting exercises) or a no intervention control condition. Experimental conditions were implemented one week after the baseline run about 20 minutes before a second one mile trial. RESULTS: Participants assigned to the motivational intervention improved their running performance significantly more than those assigned to the other two conditions. Although the magnitude of the effect was small, participants assigned to yoga exercises showed significant improvements in running performance relative to control condition participants. Consumer satisfaction ratings indicated that participants who were assigned to the motivational and yoga exercise groups liked their interventions more than those assigned to the control group. CONCLUSION: Motivational and yoga interventions designed to improve long distance running performance were equally acceptable to the participants, but the former had a greater effect. PMID- 16371494 TI - Cardiocirculatory and metabolic responses at different walking intensities. AB - OBJECTIVES: Although walking is a common physical activity, scientifically based training guidelines using standardised tests have not been established. Therefore this explorative study investigated the cardiovascular and metabolic load resulting from different walking intensities derived from maximal velocity (Vmax) during an incremental treadmill walking test. METHODS: Oxygen uptake, heart rate (HR), blood concentrations of lactate and catecholamines, and rating of perceived exertion were recorded in 16 recreational athletes (mean (SD) age 53 (9) years) during three 30 minute walking trials at 70%, 80%, and 90% of Vmax (V70, V80, and V90) attained during an incremental treadmill walking test. RESULTS: Mean (SD) oxygen uptake was 18.2 (2.3), 22.3 (3.1), and 29.3 (5.0) ml/min/kg at V70, V80, and V90 respectively (p<0.001). V70 led to a mean HR of 110 (9) beats/min (66% HRmax), V80 to 124 (9) beats/min (75% HRmax), and V90 to 152 (13) beats/min (93% HRmax) (p<0.001). Mean (SD) lactate concentrations were 1.1 (0.2), 1.8 (0.6), and 3.9 (2.0) mmol/l at V70, V80, and V90 respectively (p<0.001). There were no significant differences between catecholamine concentrations at the different intensities. Rating of perceived exertion was 10 (2) at V70, 12 (2) at V80, and 15 (2) at V90. Twelve subjects reported muscular complaints during exercise at V90 but not at V70 and V80. CONCLUSIONS: Intensity and heart rate prescriptions for walking training can be derived from an incremental treadmill walking test. The cardiovascular and metabolic reactions observed suggest that V80 is the most efficient workload for training in recreational athletes. Further studies are needed to confirm these findings. PMID- 16371495 TI - Cervical spinal injury in children's community rugby football. AB - OBJECTIVE: To examine the characteristics of cervical spinal injury (CSI) in school aged children injured in community based rugby football who presented to the emergency department for assessment. METHODS: This is a retrospective descriptive case series study reviewing the medical records of all children younger than 15 years of age who presented to the emergency department at the Children's Hospital at Westmead, Sydney, Australia for assessment of injury to the cervical spine between 2000 and 2003. RESULTS: There were 125 children with CSI; most (97%) were boys of mean age 12.7 years. Injuries occurred throughout the season, with an unexpected peak in June. Neck pain was the main presenting complaint (98%). Neurological symptoms were reported in 43%, half having concussion. Hyperextension of the neck accounted for a third of all cases and was usually the result of a spear tackle. Appropriate treatment of the cervical spine on the field of play before transport to hospital was inconsistently performed. Half of the players with CSI suffered secondary injuries, consisting of concussive head injury, faciomaxillary injury, eye injury, or limb fracture. Admission to hospital was common, with all children admitted undergoing further radiological assessment. Two minor fractures were reported and no permanent neurological disability. Overall, no adverse events were reported and the clinical outcome was good. CONCLUSION: CSI in children playing rugby football is rarely catastrophic although often associated with other injuries. Continued efforts are needed to educate players and referees to prevent injury. PMID- 16371496 TI - No cumulative effects for one or two previous concussions. AB - BACKGROUND: Sports medicine clinicians and the general public are interested in the possible cumulative effects of concussion. OBJECTIVE: To examine whether athletes with a history of one or two previous concussions differed in their preseason neuropsychological test performances or symptom reporting. METHOD: Participants were 867 male high school and university amateur athletes who completed preseason testing with ImPACT version 2.0. They were sorted into three groups on the basis of number of previous concussions. There were 664 athletes with no previous concussions, 149 with one previous concussion, and 54 with two previous concussions. Multivariate analysis of variance was conducted using the verbal memory, visual memory, reaction time, processing speed, and postconcussion symptom composite scores as dependent variables and group membership as the independent variable. RESULTS: There was no significant multivariate effect, nor were there any significant main effects for individual scores. There was no measurable effect of one or two previous concussions on athletes' preseason neuropsychological test performance or symptom reporting. CONCLUSION: If there is a cumulative effect of one or two previous concussions, it is very small and undetectable using this methodology. PMID- 16371497 TI - A randomised, placebo controlled trial of low level laser therapy for activated Achilles tendinitis with microdialysis measurement of peritendinous prostaglandin E2 concentrations. AB - BACKGROUND: Low level laser therapy (LLLT) has gained increasing popularity in the management of tendinopathy and arthritis. Results from in vitro and in vivo studies have suggested that inflammatory modulation is one of several possible biological mechanisms of LLLT action. OBJECTIVE: To investigate in situ if LLLT has an anti-inflammatory effect on activated tendinitis of the human Achilles tendon. SUBJECTS: Seven patients with bilateral Achilles tendinitis (14 tendons) who had aggravated symptoms produced by pain inducing activity immediately before the study. METHOD: Infrared (904 nm wavelength) LLLT (5.4 J per point, power density 20 mW/cm2) and placebo LLLT (0 J) were administered to both Achilles tendons in random blinded order. RESULTS: Ultrasonography Doppler measurements at baseline showed minor inflammation through increased intratendinous blood flow in all 14 tendons and measurable resistive index in eight tendons of 0.91 (95% confidence interval 0.87 to 0.95). Prostaglandin E2 concentrations were significantly reduced 75, 90, and 105 minutes after active LLLT compared with concentrations before treatment (p = 0.026) and after placebo LLLT (p = 0.009). Pressure pain threshold had increased significantly (p = 0.012) after active LLLT compared with placebo LLLT: the mean difference in the change between the groups was 0.40 kg/cm2 (95% confidence interval 0.10 to 0.70). CONCLUSION: LLLT at a dose of 5.4 J per point can reduce inflammation and pain in activated Achilles tendinitis. LLLT may therefore have potential in the management of diseases with an inflammatory component. PMID- 16371498 TI - A controlled clinical pilot trial to study the effectiveness of ice as a supplement to the exercise programme for the management of lateral elbow tendinopathy. AB - BACKGROUND: The use of ice as a supplement to an exercise programme has been recommended for the management of lateral elbow tendinopathy (LET). No studies have examined its effectiveness. OBJECTIVES: To investigate whether an exercise programme supplemented with ice is more successful than the exercise programme alone in treating patients with LET. METHODS: Patients with unilateral LET for at least four weeks were included in this pilot study. They were sequentially allocated to receive five times a week for four weeks either an exercise programme with ice or the exercise programme alone. The exercise programme consisted of slow progressive eccentric exercises of wrist extensors and static stretching of the extensor carpi radialis brevis tendon. In the exercise programme/ice group, the ice was applied after the exercise programme for 10 minutes in the form of an ice bag to the facet of the lateral epicondyle. Patients were evaluated at baseline, at the end of treatment, and three months after the end of treatment. Outcome measures used were the pain visual analogue scale and the dropout rate. RESULTS: Forty patients met the inclusion criteria. At the end of treatment there was a decline in visual analogue scale of about 7 units in both groups compared with baseline (p<0.0005, paired t test). There were no significant differences in the magnitude of reduction between the groups at the end of treatment and at the three month follow up (p<0.0005, independent t test). There were no dropouts. CONCLUSIONS: An exercise programme consisting of eccentric and static stretching exercises had reduced the pain in patients with LET at the end of the treatment and at the follow up whether or not ice was included. Further research to establish the relative, absolute, and cost effectiveness as well as the mechanism of action of the exercise programme is needed. PMID- 16371500 TI - Cerebellar hypoplasia and quadrupedal locomotion in humans as a recessive trait mapping to chromosome 17p. AB - BACKGROUND: Congenital hereditary non-progressive hypoplasia of the cerebellum is a rare condition, frequently associated with other neuropathology such as lissencephaly. Clinically, the condition is associated with variable degrees of mental retardation, microcephaly, seizures, and movement disorders due to ataxia. In severe cases, patients are unable to ambulate independently, but nevertheless do use bipedal locomotion. METHODS AND RESULTS: Here we present a family with seven affected members, five of whom never learned to walk on two legs but have fully adapted to quadrupedal palmigrade locomotion. These subjects show signs of cerebellar ataxia and are mentally retarded. MRI analysis demonstrated hypoplasia of the cerebellum and the cerebellar vermis as well as a small nucleus dentatus and a thin corpus callosum but no other malformations. We show, by a genome-wide linkage scan, that quadrupedal locomotion is a recessive trait linked to chromosome 17p. CONCLUSIONS: Our findings have implications for understanding the neural mechanism mediating bipedalism, and, perhaps, the evolution of this unique hominid trait. PMID- 16371501 TI - Letting the family know: balancing ethics and effectiveness when notifying relatives about genetic testing for a familial disorder. AB - OBJECTIVE: To increase the awareness among at risk relatives of the availability of genetic testing for a familial disorder while respecting their autonomy and privacy. METHODS: This was a comparison of preintervention and postintervention cohorts of families carried out in a state wide clinical service providing genetic counselling and testing for people at risk of familial adult onset cancer. Unaffected relatives who were not clients of the service in 74 kindreds with familial mutations causing familial breast and ovarian cancer, hereditary non-polyposis colorectal cancer, or Cowden syndrome were included in the study. In the baseline cohort (41 kindreds), family members who were clients of the clinical service and had been shown to be carriers of mutations were asked to advise relatives that genetic testing was available. In the intervention cohort (33 kindreds), the clinical service obtained consent to advise at risk relatives by letter that genetic testing was available. The main outcome measures were: (a) proportion of unaffected first and second degree relatives of the proband in each family whose genetic status was clarified within 2 years of the mutation being identified in the family, and (b) concerns regarding privacy and autonomy voiced by relatives receiving these letters. RESULTS: In the baseline cohort, the average proportion of relatives in each family whose genetic status was clarified was 23%. In the intervention cohort, the average proportion of relatives in each family whose genetic status was clarified was 40% (p = 0.001). None of the relatives in the intervention cohort complained of a breach of privacy or autonomy. CONCLUSION: Clinical services can take an effective and proactive approach to notifying relatives who are not their clients of the availability of genetic testing without compromising principles of privacy and autonomy. PMID- 16371502 TI - OTOF mutations revealed by genetic analysis of hearing loss families including a potential temperature sensitive auditory neuropathy allele. AB - INTRODUCTION: The majority of hearing loss in children can be accounted for by genetic causes. Non-syndromic hearing loss accounts for 80% of genetic hearing loss in children, with mutations in DFNB1/GJB2 being by far the most common cause. Among the second tier genetic causes of hearing loss in children are mutations in the DFNB9/OTOF gene. METHODS: In total, 65 recessive non-syndromic hearing loss families were screened by genotyping for association with the DFNB9/OTOF gene. Families with genotypes consistent with linkage or uninformative for linkage to this gene region were further screened for mutations in the 48 known coding exons of otoferlin. RESULTS: Eight OTOF pathological variants were discovered in six families. Of these, Q829X was found in two families. We also noted 23 other coding variant, believed to have no pathology. A previously published missense allele I515T was found in the heterozygous state in an individual who was observed to be temperature sensitive for the auditory neuropathy phenotype. CONCLUSIONS: Mutations in OTOF cause both profound hearing loss and a type of hearing loss where otoacoustic emissions are spared called auditory neuropathy. PMID- 16371503 TI - Rapid, diffusional shuttling of poly(A) RNA between nuclear speckles and the nucleoplasm. AB - Speckles are nuclear bodies that contain pre-mRNA splicing factors and polyadenylated RNA. Because nuclear poly(A) RNA consists of both mRNA transcripts and nucleus-restricted RNAs, we tested whether poly(A) RNA in speckles is dynamic or rather an immobile, perhaps structural, component. Fluorescein-labeled oligo(dT) was introduced into HeLa cells stably expressing a red fluorescent protein chimera of the splicing factor SC35 and allowed to hybridize. Fluorescence correlation spectroscopy (FCS) showed that the mobility of the tagged poly(A) RNA was virtually identical in both speckles and at random nucleoplasmic sites. This same result was observed in photoactivation-tracking studies in which caged fluorescein-labeled oligo(dT) was used as hybridization probe, and the rate of movement away from either a speckle or nucleoplasmic site was monitored using digital imaging microscopy after photoactivation. Furthermore, the tagged poly(A) RNA was observed to rapidly distribute throughout the entire nucleoplasm and other speckles, regardless of whether the tracking observations were initiated in a speckle or the nucleoplasm. Finally, in both FCS and photoactivation-tracking studies, a temperature reduction from 37 to 22 degrees C had no discernible effect on the behavior of poly(A) RNA in either speckles or the nucleoplasm, strongly suggesting that its movement in and out of speckles does not require metabolic energy. PMID- 16371504 TI - Involvement of Src family kinases in N-cadherin phosphorylation and beta-catenin dissociation during transendothelial migration of melanoma cells. AB - N-cadherin is recruited to the heterotypic contact during transendothelial migration of melanoma cells in a coculture system with tumor cells seeded on top of a monolayer of endothelial cells. However, beta-catenin dissociates from N cadherin and redistributes to the nucleus of transmigrating melanoma cells to activate gene transcription. In this report, we demonstrate that Src becomes activated at the heterotypic contact between the transmigrating melanoma cell and neighboring endothelial cells. Src activation shows close temporal correlation with tyrosine phosphorylation of N-cadherin. Expression of a dominant-negative Src in melanoma cells blocks N-cadherin phosphorylation, beta-catenin dissociation, and nuclear translocation in transmigrating cells, consistent with the involvement of Src family kinases. In in vitro binding assays, Src-mediated phosphorylation of the N-cadherin cytoplasmic domain results in a significant reduction in beta-catenin binding. Although five phospho-tyrosine residues can be identified on the N-cadherin cytoplasmic domain by mass spectrometry, site specific mutagenesis indicates that Tyr-860 is the critical amino acid involved in beta-catenin binding. Overexpression of N-cadherin carrying the Y860F mutation inhibits the transmigration of transfected cells across the endothelium. Together, the data suggest a novel role for tyrosine phosphorylation of N cadherin by Src family kinases in the regulation of beta-catenin association during transendothelial migration of melanoma cells. PMID- 16371505 TI - Balance between transcription and RNA degradation is vital for Saccharomyces cerevisiae mitochondria: reduced transcription rescues the phenotype of deficient RNA degradation. AB - The Saccharomyces cerevisiae SUV3 gene encodes the helicase component of the mitochondrial degradosome (mtEXO), the principal 3'-to-5' exoribonuclease of yeast mitochondria responsible for RNA turnover and surveillance. Inactivation of SUV3 (suv3Delta) causes multiple defects related to overaccumulation of aberrant transcripts and precursors, leading to a disruption of mitochondrial gene expression and loss of respiratory function. We isolated spontaneous suppressors that partially restore mitochondrial function in suv3Delta strains devoid of mitochondrial introns and found that they correspond to partial loss-of-function mutations in genes encoding the two subunits of the mitochondrial RNA polymerase (Rpo41p and Mtf1p) that severely reduce the transcription rate in mitochondria. These results show that reducing the transcription rate rescues defects in RNA turnover and demonstrates directly the vital importance of maintaining the balance between RNA synthesis and degradation. PMID- 16371506 TI - Role of a Cdc42p effector pathway in recruitment of the yeast septins to the presumptive bud site. AB - The septins are GTP-binding, filament-forming proteins that are involved in cytokinesis and other processes. In the yeast Saccharomyces cerevisiae, the septins are recruited to the presumptive bud site at the cell cortex, where they form a ring through which the bud emerges. We report here that in wild-type cells, the septins typically become detectable in the vicinity of the bud site several minutes before ring formation, but the ring itself is the first distinct structure that forms. Septin recruitment depends on activated Cdc42p but not on the normal pathway for bud-site selection. Recruitment occurs in the absence of F actin, but ring formation is delayed. Mutant phenotypes and suppression data suggest that the Cdc42p effectors Gic1p and Gic2p, previously implicated in polarization of the actin cytoskeleton, also function in septin recruitment. Two hybrid, in vitro protein binding, and coimmunoprecipitation data indicate that this role involves a direct interaction of the Gic proteins with the septin Cdc12p. PMID- 16371507 TI - Dynamic nature of cleavage bodies and their spatial relationship to DDX1 bodies, Cajal bodies, and gems. AB - DDX1 bodies, cleavage bodies, Cajal bodies (CBs), and gems are nuclear suborganelles that contain factors involved in RNA transcription and/or processing. Although all four nuclear bodies can exist as distinct entities, they often colocalize or overlap with each other. To better understand the relationship between these four nuclear bodies, we examined their spatial distribution as a function of the cell cycle. Here, we report that whereas DDX1 bodies, CBs and gems are present throughout interphase, CPSF-100-containing cleavage bodies are predominantly found during S and G2 phases, whereas CstF-64 containing cleavage bodies are primarily observed during S phase. All four nuclear bodies associate with each other during S phase, with cleavage bodies colocalizing with DDX1 bodies, and cleavage bodies/DDX1 bodies residing adjacent to gems and CBs. Although inhibitors of RNA transcription had no effect on DDX1 bodies or cleavage bodies, inhibitors of DNA replication resulted in loss of CstF 64-containing cleavage bodies. A striking effect on nuclear structures was observed with latrunculin B, an inhibitor of actin polymerization, resulting in the formation of needlelike nuclear spicules made up of CstF-64, CPSF-100, RNA, and RNA polymerase II. Our results suggest that cleavage body components are highly dynamic in nature. PMID- 16371509 TI - Ena/VASP proteins can regulate distinct modes of actin organization at cadherin adhesive contacts. AB - Functional interactions between classical cadherins and the actin cytoskeleton involve diverse actin activities, including filament nucleation, cross-linking, and bundling. In this report, we explored the capacity of Ena/VASP proteins to regulate the actin cytoskeleton at cadherin-adhesive contacts. We extended the observation that Ena/vasodilator-stimulated phosphoprotein (VASP) proteins localize at cell-cell contacts to demonstrate that E-cadherin homophilic ligation is sufficient to recruit Mena to adhesion sites. Ena/VASP activity was necessary both for F-actin accumulation and assembly at cell-cell contacts. Moreover, we identified two distinct pools of Mena within individual homophilic adhesions that cells made when they adhered to cadherin-coated substrata. These Mena pools localized with Arp2/3-driven cellular protrusions as well as at the tips of cadherin-based actin bundles. Importantly, Ena/VASP activity was necessary for both modes of actin activity to be expressed. Moreover, selective depletion of Ena/VASP proteins from the tips of cadherin-based bundles perturbed the bundles without affecting the protrusive F-actin pool. We propose that Ena/VASP proteins may serve as higher order regulators of the cytoskeleton at cadherin contacts through their ability to modulate distinct modes of actin organization at those contacts. PMID- 16371508 TI - Shiga toxin regulates its entry in a Syk-dependent manner. AB - Shiga toxin (Stx) is composed of an A-moiety that inhibits protein synthesis after translocation into the cytosol, and a B-moiety that binds to Gb3 at the cell surface and mediates endocytosis of the toxin. After endocytosis, Stx is transported retrogradely to the endoplasmic reticulum, and then the A-fragment enters the cytosol. In this study, we have investigated whether toxin-induced signaling is involved in its entry. Stx was found to activate Syk and induce rapid tyrosine phosphorylation of several proteins, one protein being clathrin heavy chain. Toxin-induced clathrin phosphorylation required Syk activity, and in cells overexpressing Syk, a complex containing clathrin and Syk could be demonstrated. Depletion of Syk by small interfering RNA, expression of a dominant negative Syk mutant (Syk KD), or treatment with the Syk inhibitor piceatannol inhibited not only Stx-induced clathrin phosphorylation but also endocytosis of the toxin. Also, Golgi transport of Stx was inhibited under all these conditions. In conclusion, our data suggest that Stx regulates its entry into target cells. PMID- 16371510 TI - Microtubule regulation in mitosis: tubulin phosphorylation by the cyclin dependent kinase Cdk1. AB - The activation of the cyclin-dependent kinase Cdk1 at the transition from interphase to mitosis induces important changes in microtubule dynamics. Cdk1 phosphorylates a number of microtubule- or tubulin-binding proteins but, hitherto, tubulin itself has not been detected as a Cdk1 substrate. Here we show that Cdk1 phosphorylates beta-tubulin both in vitro and in vivo. Phosphorylation occurs on Ser172 of beta-tubulin, a site that is well conserved in evolution. Using a phosphopeptide antibody, we find that a fraction of the cell tubulin is phosphorylated during mitosis, and this tubulin phosphorylation is inhibited by the Cdk1 inhibitor roscovitine. In mitotic cells, phosphorylated tubulin is excluded from microtubules, being present in the soluble tubulin fraction. Consistent with this distribution in cells, the incorporation of Cdk1 phosphorylated tubulin into growing microtubules is impaired in vitro. Additionally, EGFP-beta3-tubulin(S172D/E) mutants that mimic phosphorylated tubulin are unable to incorporate into microtubules when expressed in cells. Modeling shows that the presence of a phosphoserine at position 172 may impair both GTP binding to beta-tubulin and interactions between tubulin dimers. These data indicate that phosphorylation of tubulin by Cdk1 could be involved in the regulation of microtubule dynamics during mitosis. PMID- 16371511 TI - The slow Wallerian degeneration protein, WldS, binds directly to VCP/p97 and partially redistributes it within the nucleus. AB - Slow Wallerian degeneration (Wld(S)) mutant mice express a chimeric nuclear protein that protects sick or injured axons from degeneration. The C-terminal region, derived from NAD(+) synthesizing enzyme Nmnat1, is reported to confer neuroprotection in vitro. However, an additional role for the N-terminal 70 amino acids (N70), derived from multiubiquitination factor Ube4b, has not been excluded. In wild-type Ube4b, N70 is part of a sequence essential for ubiquitination activity but its role is not understood. We report direct binding of N70 to valosin-containing protein (VCP; p97/Cdc48), a protein with diverse cellular roles including a pivotal role in the ubiquitin proteasome system. Interaction with Wld(S) targets VCP to discrete intranuclear foci where ubiquitin epitopes can also accumulate. Wld(S) lacking its N-terminal 16 amino acids (N16) neither binds nor redistributes VCP, but continues to accumulate in intranuclear foci, targeting its intrinsic NAD(+) synthesis activity to these same foci. Wild type Ube4b also requires N16 to bind VCP, despite a more C-terminal binding site in invertebrate orthologues. We conclude that N-terminal sequences of Wld(S) protein influence the intranuclear location of both ubiquitin proteasome and NAD(+) synthesis machinery and that an evolutionary recent sequence mediates binding of mammalian Ube4b to VCP. PMID- 16371512 TI - Barrier-to-autointegration factor phosphorylation on Ser-4 regulates emerin binding to lamin A in vitro and emerin localization in vivo. AB - Barrier-to-autointegration factor (BAF) is a conserved 10-kDa chromatin protein essential in proliferating cells. BAF dimers bind double-stranded DNA, histone H3, histone H1.1, lamin A, and transcription regulators, plus emerin and other LEM-domain nuclear proteins. Two-dimensional gel analysis showed that endogenous human and Xenopus BAF are posttranslationally modified by phosphorylation and potentially other modifications and that they are hyperphosphorylated during mitosis. The invariant Ser-4 residue on BAF is a major site of phosphorylation during both interphase and mitosis. In HeLa cells that overexpressed the phosphomimetic BAF missense mutant S4E, but not S4A, emerin mislocalized from the nuclear envelope, suggesting Ser-4-nonphosphorylated BAF normally promotes emerin localization at the nuclear envelope. Supporting this model, wild-type BAF but not mutant S4E enhanced emerin binding to lamin A in vitro. Thus, Ser-4 unphosphorylated BAF has a positive role in localizing emerin; this role may be disease relevant because loss or mislocalization of emerin causes Emery-Dreifuss muscular dystrophy. Our findings further suggest Ser-4 phosphorylation inhibits BAF binding to emerin and lamin A, and thereby weakens emerin-lamin interactions during both mitosis and interphase. PMID- 16371513 TI - Re: "Cancer mortality among US men and women with asthma and hay fever". PMID- 16371514 TI - Cause-specific mortality of grand multiparous women in Finland. AB - Knowledge is limited on mortality of grand multiparous women (> or =5 deliveries), whose hormonal, metabolic, and social conditions differ from the average. The authors studied overall and cause-specific mortality in 1974-2001 among 87,922 grand multiparous women including 3,678 grand grand multiparous women (> or =10 deliveries) in Finland. Standardized mortality ratios were defined as ratios of observed to expected numbers of deaths, both derived from national cause-of-death files. During follow-up, 18,870 grand multiparous women and 625 grand grand multiparous women died (standardized mortality ratios (SMRs) = 0.95 and 1.01, respectively). Decreased mortality among grand multiparous women was found for cancers of the breast (SMR = 0.64, 95% confidence interval (CI): 0.59, 0.69), corpus uteri (SMR = 0.68, 95% CI: 0.56, 0.80), ovary (SMR = 0.68, 95% CI: 0.60, 0.75), bladder (SMR = 0.59, 95% CI: 0.41, 0.82), and respiratory tract (SMR = 0.80, 95% CI: 0.72, 0.88). The only malignant tumor associated with elevated mortality was kidney cancer (SMR = 1.38, 95% CI: 1.21, 1.56). The standardized mortality ratio was also low for dementia (SMR = 0.78, 95% CI: 0.72, 0.84), respiratory diseases (SMR = 0.80, 95% CI: 0.75, 0.85), and accidents and violent causes (SMR = 0.79, 95% CI: 0.73, 0.84). Mortality from diabetes mellitus (SMR = 1.42, 95% CI: 1.29, 1.55) and ischemic heart disease (SMR = 1.10, 95% CI: 1.08, 1.13) was increased. According to this study, overall mortality among grand multiparous women is not elevated. Low mortality from cancers is offset by higher mortality from cardiovascular conditions and diabetes mellitus. PMID- 16371515 TI - Results of multivariable logistic regression, propensity matching, propensity adjustment, and propensity-based weighting under conditions of nonuniform effect. AB - Observational studies often provide the only available information about treatment effects. Control of confounding, however, remains challenging. The authors compared five methods for evaluating the effect of tissue plasminogen activator on death among 6,269 ischemic stroke patients registered in a German stroke registry: multivariable logistic regression, propensity score-matched analysis, regression adjustment with the propensity score, and two propensity score-based weighted methods-one estimating the treatment effect in the entire study population (inverse-probability-of-treatment weights), another in the treated population (standardized-mortality-ratio weights). Between 2000 and 2001, 212 patients received tissue plasminogen activator. The crude odds ratio between tissue plasminogen activator and death was 3.35 (95% confidence interval: 2.28, 4.91). The adjusted odds ratio depended strongly on the adjustment method, ranging from 1.11 (95% confidence interval: 0.67, 1.84) for the standardized mortality-ratio weighted to 10.77 (95% confidence interval: 2.47, 47.04) for the inverse-probability-of-treatment-weighted analysis. For treated patients with a low propensity score, risks of dying were high. Exclusion of patients with a propensity score of <5% yielded comparable odds ratios of approximately 1 for all methods. High levels of nonuniform treatment effect render summary estimates very sensitive to the weighting system explicit or implicit in an adjustment technique. Researchers need to be clear about the population for which an overall treatment estimate is most suitable. PMID- 16371516 TI - Effect of highly active antiretroviral therapy on multiple AIDS-defining illnesses among male HIV seroconverters. AB - The effect of highly active antiretroviral therapy (HAART) on multiple acquired immunodeficiency syndrome (AIDS)-defining illnesses remains unclear. Between 1984 and 2005, 573 male human immunodeficiency virus seroconverters in four US urban centers were followed for a median of 9.7 years. During follow-up, 345, 113, 50, and 65 men incurred 0, 1, 2, and >2 AIDS-defining illnesses, respectively. The authors extend the Cox proportional hazards model to determine whether the effect of HAART, as measured by calendar periods, persists beyond the first AIDS defining illness. After adjustment for race and age at seroconversion, the hazards of a first through third AIDS-defining illness in the HAART calendar period (beyond July 1995) were 0.31 (95% confidence interval (CI): 0.21, 0.46), 0.39 (95% CI: 0.22, 0.74), and 0.33 (95% CI: 0.14, 0.79), respectively, relative to the monotherapy and combination therapy reference calendar period (January 1990-July 1995) and therefore did not attenuate with the number of prior AIDS defining illnesses (p for homogeneity = 0.83). After the authors averaged over multiple AIDS-defining illnesses, the hazard of an AIDS-defining illness in the HAART calendar period was 0.34 (95% CI: 0.25, 0.45) relative to the reference calendar period. HAART protects against initial and subsequent AIDS-defining illnesses, whose inclusion in analysis markedly increased the precision of the estimated hazard ratio. PMID- 16371517 TI - Norman Harold Horowitz, 1915-2005. PMID- 16371518 TI - Random genetic drift and gamete frequency. AB - An analytic expression of conditional expectation of transient gamete frequency, given that one of the two loci remains polymorphic, is obtained in terms of the diffusion process by calculating the moments of the distribution. Using this expression, a model where linkage disequilibrium is introduced by a single mutation is considered. The conditional expectation of the gamete frequency given that the locus with the mutant allele remains polymorphic is presented. The behavior is significantly different from the monotonic decrease observed in the deterministic model without random genetic drift. PMID- 16371519 TI - Development of a research agenda for endovascular treatment of venous thromboembolism: proceedings from a multidisciplinary consensus panel. PMID- 16371520 TI - Development of a research agenda for endovenous treatment of lower-extremity venous reflux: proceedings from a multidisciplinary consensus panel. PMID- 16371521 TI - Pain and anxiety: two problems, two solutions. PMID- 16371522 TI - Pain and anxiety during interventional radiologic procedures: effect of patients' state anxiety at baseline and modulation by nonpharmacologic analgesia adjuncts. AB - PURPOSE: To assess how patients' underlying anxiety affects their experience of distress, use of resources, and responsiveness toward nonpharmacologic analgesia adjunct therapies during invasive procedures. MATERIALS AND METHODS: Two hundred thirty-six patients undergoing vascular and renal interventions, who had been randomized to receive during standard care treatment, structured empathic attention, or self-hypnotic relaxation, were divided into two groups: those with low state anxiety scores on the State-Trait Anxiety Inventory (STAI, scores < 43; n = 116) and those with high state anxiety scores (> or = 43; n = 120). All had access to patient-controlled analgesia with fentanyl and midazolam. Every 15 minutes during the procedure, patients rated their anxiety and pain on a scale of 0-10 (0, no pain/anxiety at all; 10, worst possible pain/anxiety). Effects were assessed by analysis of variance and repeated-measures analysis. RESULTS: Patients with high state anxiety levels required significantly greater procedure time and medication. Empathic attention as well as hypnosis treatment reduced procedure time and medication use for all patients. These nonpharmacologic analgesia adjunct treatments also provided significantly better pain control than standard care for patients with low anxiety levels. Anxiety decreased over the time of the procedure; patients with high state anxiety levels experienced the most significant decreases in anxiety with nonpharmacologic adjuncts whereas patients with low state anxiety levels coped relatively well under all conditions. CONCLUSION: Patients' state anxiety level is a predictor of trends in procedural pain and anxiety, need for medication, and procedure duration. Low and high state anxiety groups profit from the use of nonpharmacologic analgesia adjuncts but those with high state anxiety levels have the most to gain. PMID- 16371523 TI - Use of the peripheral cutting balloon to treat hemodialysis-related stenoses. AB - PURPOSE: To compare the effectiveness and safety of use of the peripheral cutting balloon (PCB) versus standard percutaneous transluminal angioplasty (PTA) for the treatment of hemodialysis-related stenoses. MATERIALS AND METHODS: This prospective, randomized multicenter clinical trial included 340 patients with stenotic or thrombosed hemodialysis grafts who were randomized to receive treatment with the PCB or PTA for venous outflow stenosis. One hundred seventy three patients underwent treatment with the PCB, 101 with stenotic grafts and 72 with thrombosed grafts. PTA was used to treat 167 patients, 94 patients with stenotic grafts and 73 with thrombosed grafts. The follow-up period extended for 6 months. RESULTS: The procedural success rates were 80.8% and 75.4% for the PCB and PTA groups, respectively (P = .24). With use of the PCB, the primary patency rates of the target lesions were 84.3%, 65.8%, and 47.9% at 1 month, 3 months, and 6 months, respectively. With PTA, the primary patency rates of the target lesions were 77.7%, 63.4%, and 40.5% at 1 month, 3 months, and 6 months, respectively. The primary patency rates of the entire vascular access circuit were 82.6%, 61.0%, and 43.3% at 1 month, 3 months, and 6 months, respectively, with use of the PCB. For patients who were treated with PTA, the primary patency rates of the vascular access circuit were 75.9%, 61.0%, and 36.3% at 1 month, 3 months, and 6 months, respectively. When comparing the PCB and PTA, there was no difference in the 6-month primary patency rates in the target lesion (P = .373) or the entire vascular access circuit (P = .531). There were nine device-related complications in the PCB group (5.2%): five venous ruptures (2.9%), three venous dissections (1.7%), and one case of thrombosis (0.6%). There were no device related complications in the PTA group. CONCLUSION: This prospective, randomized trial comparing use of the PCB versus standard PTA for treatment of hemodialysis related venous stenoses demonstrated that the PCB provides equivalent 6-month patency to PTA for stenotic and thrombosed grafts. PMID- 16371525 TI - Single-center experience with the Arrow-Trerotola Percutaneous Thrombectomy Device in the management of thrombosed native dialysis fistulas. AB - PURPOSE: The present study sought to evaluate the performance of the Arrow Trerotola Percutaneous Thrombolytic Device (PTD) in the treatment of native fistula thrombosis in a U. S. hemodialysis population. Specifically, the technical success, clinical success, complication rate and type, primary and secondary patency rates, effect of adjunctive thrombolytic therapy, and any variables that affected outcomes of procedures in which this device was used were analyzed. MATERIALS AND METHODS: Forty-two patients with 44 thrombosed native fistulas (17 radiocephalic, 10 brachiocephalic, 10 transposed or superficialized, five graft/fistula hybrids, and two leg fistulas) were treated with 62 mechanical thrombolysis procedures with use of the PTD. All patients had large clot burden. The device type was recorded in 43 procedures: standard (n = 21), over-the-wire (OTW; n = 19), or both (n = 3). No device was used in two cases because of inability to cross the anastomosis. Adjunctive therapies (n = 18) included the use of tissue plasminogen activator (tPA; n = 16) and deployment of the AngioJet device with (n = 1) or without tPA (n = 1). Stents were inserted in four procedures. Outcome variables included technical and clinical success, complications, and primary and secondary patency. Cox proportional-hazards regression and Kaplan-Meier analyses were performed. RESULTS: The technical success rate was 87% (54 of 62) and the clinical success rate was 79% (49 of 62). Percutaneous transluminal angioplasty was performed in all but two procedures. Complications occurred in 13% of procedures (n = 8); three resulted in technical failure. The primary patency rates were 38% at 6 months and 18% at 12 months; secondary patency rates were 74% and 69%, respectively. Outcomes were not affected by adjunctive techniques, fistula type, age of fistula, device type (ie, OTW vs standard), or patient sex. Secondary patency was superior when no residual clot or stenosis was present (P = .003). CONCLUSIONS: The PTD is effective for percutaneous treatment of thrombosed hemodialysis fistulas, with good short- and long-term outcomes in a U.S. population. Within the limitations of a retrospective study with a small sample size, use of an adjunctive thrombolytic agent did not appear to improve results compared with the use of the device alone. PMID- 16371527 TI - Prospective study of balloon inflation pressures and other technical aspects of hemodialysis access angioplasty. AB - PURPOSE: Balloon angioplasty is a mainstay in the treatment of failing or thrombosed hemodialysis access grafts and fistulas. A sizable body of outcomes data exists concerning percutaneous transluminal angioplasty (PTA) in hemodialysis access, yet there is a relative paucity of technical information available, especially regarding dilation pressures. The aim of the present study was to compile such information, which can be critical to the choice of devices for PTA and to the design of future clinical trials seeking to improve outcomes in this area. MATERIALS AND METHODS: Technical data were collected prospectively for 102 PTA procedures (66 prophylactic PTA procedures and 36 PTA procedures performed during access thrombectomy). Demographic data concerning the access were collected. Technical data were collected individually for each lesion treated, including lesion location; degree of stenosis (in quartiles); lesion length; PTA balloon brand, size, and length; pressure at which the waist of the balloon was effaced; residual stenosis; and reason for additional balloons or inflations if used. Outcomes data other than residual stenosis were not collected, but the endpoint for all interventions was a thrill in the access. RESULTS: A total of 230 lesions were treated. Two (1%) could not be successfully treated with PTA despite the use of "ultra high" pressure (approximately 40 atm); one was treated successfully with parallel wire technique and the other was revised surgically after the use of a cutting balloon also failed. Overall, 55% of lesions required pressures greater than 15 atm to efface the waist. Excluding initial failures, 20% of lesions in native fistulas and 9% in grafts required very high pressure (>20 atm) to efface the waist (P = .02). High pressure was needed less frequently in PTA procedures performed in the setting of thrombectomy procedures than in prophylactic PTA procedures (P = .0001). Residual stenosis was positively correlated with severity of initial stenosis and negatively correlated with duration of inflation. CONCLUSIONS: Conventional angioplasty balloons are inadequate for the treatment of most hemodialysis access stenoses. High pressures (>15 atm) are commonly needed for PTA in hemodialysis access. Very high pressures (>20 atm) are more frequently needed in native fistulas. PMID- 16371528 TI - Comparison of SMART stent placement for arteriovenous graft salvage versus successful graft PTA. AB - PURPOSE: To compare the SMART (shape memory alloy recoverable technology) stent with percutaneous transluminal angioplasty (PTA) alone in hemodialysis access venous stenoses. MATERIALS AND METHODS: A prospective, nonrandomized study was undertaken in 60 patients with dysfunctional polytetrafluoroethylene dialysis grafts. Indications for stent placement were acute PTA failure, rapid restenosis, and vessel perforation. The primary endpoint was improved graft patency in patients treated with stents compared with that in patients whose disease responded to PTA alone. The secondary endpoints were lower postprocedural midgraft pressures and similar complication rates compared with PTA alone. RESULTS: The key venous stenosis was at the graft-to-vein anastomosis in all but two patients. Thirty-five patients showed a response to PTA alone. Sixteen patients received stents for stenoses greater than 30% after angioplasty, six for rapidly recurrent stenosis, and three for venous rupture. Nine patients received stents across the level of the elbow joint. Stenosis after intervention was significantly less frequent in the stent group (7% vs 16%; P = .001), but the midgraft systolic pressure ratios did not significantly differ. The clinical success rates were 100% after stent implantation and 97% after PTA alone. Except for venous rupture, there were no procedure-related complications, and, excluding early graft thrombosis, there were no complications at 30 days. A single stent fracture was found on follow-up. The mean primary graft patency times were 5.6 months after PTA and 8.2 months after stent treatment (P = .050). When stents were placed across the level of the elbow joint, the mean primary graft patency time was 8.9 months. CONCLUSION: Implantation of the SMART stent is safe and effective for the treatment of residual or rapidly recurrent dialysis access venous stenoses and is associated with better patency than PTA alone. PMID- 16371529 TI - Treatment of unresectable hepatocellular carcinoma with use of 90Y microspheres (TheraSphere): safety, tumor response, and survival. AB - PURPOSE: To present safety and efficacy results obtained in treatment of a cohort of patients with unresectable hepatocellular carcinoma (HCC) with use of 90Y microspheres (TheraSphere). PATIENTS AND METHODS: Forty-three consecutive patients with HCC were treated with 90Y microspheres over a 4-year period. Patients were treated by liver segment or lobe on one or more occasions based on tumor distribution, liver function, and vascular flow dynamics. Patients were followed for adverse events, objective tumor response, and survival. Patients were stratified into three risk groups according to method of treatment and risk stratification (group 0, segmental; group 1, lobar low-risk; group 2, lobar high risk) and Okuda and Child-Pugh scoring systems. RESULTS: Based on follow-up data from 43 treated patients, 20 patients (47%) had an objective tumor response based on percent reduction in tumor size and 34 patients (79%) had a tumor response when percent reduction and/or tumor necrosis were used as a composite measure of tumor response. There was no statistical difference among the three risk groups with respect to tumor response. Survival times from date of diagnosis were different among the risk groups (P < .0001). Median survival times were 46.5 months, 16.9 months, and 11.1 months for groups 0, 1, and 2, respectively. Median survival times of 24.4 months and 12.5 months by Okuda scores of I and II, respectively, were achieved (mean, 25.8 months vs 13.1). Patients had median survival times of 20.5 months and 13.8 months according to Child class A and class B/C disease, respectively (mean, 22.7 months vs 13.6 months). Patients classified as having diffuse disease exhibited decreased survival and reduced tumor response. There were no life-threatening adverse events related to treatment. CONCLUSIONS: Use of 90Y microspheres (TheraSpheres) provides a safe and effective method of treatment for a broad spectrum of patients presenting with unresectable HCC. Further investigation is warranted. PMID- 16371530 TI - 90Y microsphere (TheraSphere) treatment for unresectable colorectal cancer metastases of the liver: response to treatment at targeted doses of 135-150 Gy as measured by [18F]fluorodeoxyglucose positron emission tomography and computed tomographic imaging. AB - PURPOSE: The purpose of this phase II study was to determine the safety and efficacy of TheraSphere treatment (90Y microspheres) in patients with liver dominant colorectal metastases in whom standard therapies had failed or were judged to be inappropriate. MATERIALS AND METHODS: Twenty-seven patients with unresectable hepatic colorectal metastases were treated at a targeted absorbed dose of 135-150 Gy. Safety and toxicity were assessed according to the National Cancer Institute's Common Toxicity Criteria, version 3.0. Response was assessed with use of computed tomography (CT) and was correlated with response on [18F]fluorodeoxyglucose (FDG) positron emission tomography (PET). Survival from first treatment was estimated with use of the Kaplan-Meier method. RESULTS: Tumor response measured by FDG PET imaging exceeded that measured by CT imaging for the first (88% vs 35%) and second (73% vs 36%) treated lobes. Tumor replacement of 25% or less (vs >25%) was associated with a statistically significant increase in median survival (339 days vs 162 days; P = .002). Treatment-related toxicities included mild fatigue (n = 13; 48%), nausea (n = 4; 15%), and vague abdominal pain (n = 5; 19%). There was one case of radiation-induced gastritis from inadvertent deposition of microspheres to the gastrointestinal tract (n = 1; 4%). Three patients (11%) experienced ascites/pleural effusion after treatment with TheraSphere as a consequence of liver failure in advanced-stage metastatic disease. With the exception of these three patients whose sequelae were not considered to be related to treatment, all observed toxicities were transient and resolved without medical intervention. CONCLUSION: TheraSphere administration appears to provide stabilization of liver disease with minimal toxicity in patients in whom standard systemic chemotherapy regimens have failed. PMID- 16371532 TI - Safety and efficacy of transarterial chemoembolization in patients with unresectable hepatocellular carcinoma and portal vein thrombosis. AB - PURPOSE: Despite the absence of conclusive data, portal vein (PV) thrombosis is considered a contraindication to transarterial chemoembolization (TACE) in patients with unresectable hepatocellular carcinoma (HCC). The purpose of our study was to establish the safety of TACE in such patients and identify key prognostic factors and survival. MATERIALS AND METHODS: Data were prospectively collected from 32 consecutive patients with unresectable HCC and PV thrombosis who underwent treatment with TACE. History and physical examination, relevant laboratory values, and contrast material-enhanced magnetic resonance (MR) images were obtained before each TACE procedure. Repeated TACE was performed every 6 weeks unless patients developed a contraindication or MR imaging showed complete response. RESULTS: Median overall survival was 9.5 months (range, 3-50 months). Child-Pugh numerical disease stage was the prognostic factor most strongly related to survival. The 30-day mortality rate was zero and there was no evidence of TACE-related hepatic infarction or acute liver failure. The 6-, 9-, 12-, and 18-month survival rates were 60%, 47%, 25%, and 12.5%, respectively. CONCLUSIONS: PV thrombosis should not be considered a contraindication to TACE. Compared with historical control subjects who received traditional forms of treatment, the patients in the present study had extended survival. However, prospective randomized trials are necessary to show this conclusively and to show which subgroups benefit. PMID- 16371533 TI - Hepatic arterial chemoembolization for hepatocellular carcinoma: comparison of survival rates with different embolic agents. AB - PURPOSE: The optimal embolic agent for transhepatic arterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) has not been identified. This study reports outcomes of TACE for HCC with Gelfoam powder and polyvinyl alcohol (PVA). MATERIALS AND METHODS: Eighty-one patients underwent 152 TACE sessions with Gelfoam powder (n = 41) or polyvinyl alcohol (PVA) and Ethiodol (n = 40) as the embolic agent. Chemotherapeutic drugs were the same for all patients (50 mg cisplatin, 20 mg doxorubicin, 10 mg mitomycin-c). The groups were compared based on number of TACE sessions, maximum tumor size, bilirubin level, aspartate and alanine aminotransferase levels, Child-Pugh score, Model for End-stage Liver Disease score, and hepatitis B or C virus positivity. The number of cases of each Child class in each group was also evaluated. Survival starting from the first TACE session was calculated according to Kaplan-Meier analysis. Forty-eight patients died during the study period, 19 received transplants, and 14 were alive at the end of the study period. RESULTS: The groups were statistically similar in all categories regarding liver function, Child-Pugh score, tumor size, hepatitis status, and percentage of patients with Child class A, B, and C disease. The number of TACE sessions was significantly greater for the Gelfoam powder group (mean, 2.2) versus the PVA group (mean, 1.6; P = .01). Overall survival was similar between groups whether patients who received transplants were included in the analysis (mean, 659 days +/- 83 with Gelfoam powder vs 565 days +/- 71 with PVA; P = .42) or were excluded (mean, 519 days +/- 80 with Gelfoam powder vs 511 days +/- 75 with PVA; P = .93). CONCLUSION: In similar patient groups, survival after treatment of HCC with TACE with Gelfoam powder or PVA and Ethiodol was similar. PMID- 16371534 TI - Portal vein embolization with N-butyl cyanoacrylate before partial hepatectomy in patients with hepatocellular carcinoma and underlying cirrhosis or advanced fibrosis. AB - PURPOSE: To describe the safety, complications, and liver regeneration associated with the left liver after embolization of the right portal vein (PV) in patients with hepatocellular carcinoma (HCC) developed in the setting of advanced liver fibrosis and cirrhosis. MATERIALS AND METHODS: Forty patients (31 men, nine women; mean age, 62 years) with HCC underwent PV embolization over a 4-year period. Embolization was performed from a left PV percutaneous access with use of n-butyl cyanoacrylate (NBCA) mixed with iodized oil. Computed tomography (CT) volumetry was performed before and 1 month after PV embolization to measure the left lobe volume as well as the functional liver ratio defined by the ratio between the left lobe and the total liver volume minus tumoral volume. PV pressure and liver enzyme levels were compared before and 1 month after the procedure and complications were registered. Factors potentially affecting regeneration (age, sex, diabetes, chemoembolization, functional liver ratio before PV embolization, and Knodell histologic score) were evaluated by one-way and stepwise regression analysis. RESULTS: PV embolization could be achieved successfully in all cases. Two patients had partial PV thrombosis on the 1-month follow-up CT and two patients developed transient ascites after PV embolization. The left lobe volume increase was 41% +/- 32% after PV embolization and the functional liver ratio increased from 28% +/- 10% to 36% +/- 10% (P < .0001). Hypertrophy of the left lobe was greater in patients with a low functional liver ratio before PV embolization and those with an F3 fibrosis score. Other factors had no influence on left lobe regeneration. CONCLUSION: PV embolization with use of NBCA is feasible in patients with advanced fibrosis and cirrhosis. Hypertrophy of the left lobe of the liver after PV embolization has a statistically significant correlation with lower functional liver ratio and lower degrees of fibrosis. PMID- 16371535 TI - Reperfusion of pulmonary arteriovenous malformations after embolotherapy. AB - PURPOSE: To describe the mechanisms and risk factors associated with reperfusion of successfully treated pulmonary arteriovenous malformations (PAVMs) after embolotherapy. MATERIALS AND METHODS: Among 112 consecutive patients with PAVMs treated by embolotherapy, 19 patients were identified who had 33 angiographically confirmed reperfused PAVMs. A retrospective analysis of computed tomography (CT) and angiography was performed in patients with documented reperfused PAVMs in which reperfused PAVMs were compared with nonreperfused PAVMs. CT images were examined for persistence of the aneurysm and/or draining vein after initial embolotherapy and correlated with angiography to determine the mechanism of reperfusion. PAVM and embolic agent characteristics (eg, feeding artery size and number; PAVM location; coil size, number, and location) were evaluated for association with reperfusion. The outcomes of repeat embolotherapy for reperfused PAVMs were evaluated. RESULTS: The PAVM aneurysm and/or draining vein persisted on CT after initial embolotherapy in all reperfused PAVMs and resolved in all nonreperfused PAVMs (in patients with nondiffuse PAVMs). Recanalization was the mechanism of reperfusion in 88%. Reperfusion was associated with the use of a single coil (P < .0001), oversized coils (P < .0001), coil placement more than 1 cm from the aneurysm (P < .0001), and increased feeding artery size (P < .001). Repeat embolotherapy for reperfused PAVMs was technically successful in 94% of cases. In the remaining 6% of cases, insufficient feeding artery length prevented safe repeat treatment. After a mean follow-up of 41 months, 42% of reperfused PAVMs in our series have been successfully treated again and occluded. CONCLUSIONS: Recanalization is the most common mechanism of PAVM reperfusion. Increased feeding artery diameter, low number of coils, use of oversized coils, and proximal coil placement within the feeding artery are associated with reperfusion. Distal coil placement facilitates repeat embolization if reperfusion occurs. PMID- 16371536 TI - Transhepatic catheter-directed thrombectomy and thrombolysis of acute superior mesenteric venous thrombosis. AB - PURPOSE: To evaluate clinical outcomes after percutaneous treatment of superior mesenteric vein (SMV) thrombosis. MATERIALS AND METHODS: A retrospective chart review was conducted of all patients with SMV thrombosis treated with percutaneous catheter-directed thrombectomy/thrombolysis. The demographics of the study population, potential causative factors contributing to SMV thrombosis, and morbidity and mortality associated with therapy were assessed. RESULTS: Eleven patients (mean age, 44.3 years +/- 12.8) with SMV thrombosis were treated with percutaneous transhepatic catheter-directed thrombectomy/thrombolysis. Potential causative factors included recent major abdominal surgery, thrombophilic conditions, pancreatitis, and repetitive abdominal trauma. The mean duration between the onset of symptoms and percutaneous treatment was 8.6 days +/- 6.5. Computed tomography confirmed the clinical diagnosis in nine patients (81.8%). One patient (9.1%) had a bleeding complication, which was treated by chest tube drainage without long-term sequelae. One patient (9.1%) with refractory SMV thrombosis died of sepsis and multiple organ failure. No recurrent episode of SMV thrombosis or mortality was documented during a mean follow-up of 42 months +/- 22.5. CONCLUSIONS: Percutaneous transhepatic catheter-directed thrombectomy/thrombolysis for SMV thrombosis is associated with a rapid improvement in symptoms and low incidences of long-term morbidity and mortality. Percutaneous thrombectomy and thrombolysis should be considered in all patients with acute SMV thrombosis without evidence of bowel necrosis. PMID- 16371537 TI - Feasibility, safety, and efficacy of percutaneous transhepatic injection of beta cell grafts. AB - PURPOSE: To evaluate the safety, feasibility, and clinical efficacy of percutaneous transhepatic injection of beta-cell grafts in patients with type 1 diabetes mellitus. MATERIALS AND METHODS: Between December 2001 and November 2003, 15 patients with C-peptide-negative type 1 diabetes underwent 31 percutaneous injections for intraportal implantation of beta-cell grafts. Grafts consisted of cultured beta-cell preparations as previously described. In 13 cases, the transplant procedure was done under sedation, whereas in 18 cases, general anesthesia was given. In all procedures, percutaneous access to the right portal vein occurred under ultrasound (US) guidance with use of a microbore puncture needle. The subsequent catheterization of the main portal vein was performed under fluoroscopic and angiographic control with use of a microbore delivery catheter and guide wire. Clinical, biochemical, and radiologic evaluation was performed before and after the procedure. RESULTS: In all cases, it was possible to access the portal vein (median number of needle passes, 1; range, 1-6). The volume of cultured beta-cell grafts injected for each transplantation averaged 0.58 mL (range, 0.26-1.60 mL) and the mean recorded procedure time (from puncture to catheter withdrawal) was 19 minutes (range, 10 80 min). Three patients presented with transient abdominal pain immediately after the procedure; postprocedural duplex US of the liver revealed a patent portal vein and end branches in all cases and a minor perihepatic fluid collection in another three patients. From the end of week 1 to week 3, a mean 3.8-fold increase in liver aminotransferase levels was measured in all recipients after the first implantation session. A similar increase was seen in only one patient after a second transplantation session. At 6 months after transplantation, 13 of 15 patients (86%) had a functioning graft with plasma C-peptide levels greater than 0.5 ng/mL. CONCLUSIONS: The combined US, fluoroscopic, and angiographic monitoring of percutaneous transhepatic injection with use of a microbore delivery catheter is a safe and reproducible radiologic procedure for transplantation of beta-cell grafts in diabetic patients. Increased posttransplantation C-peptide levels, which demonstrate acceptable graft function, can be obtained. PMID- 16371538 TI - Fluoroscopically guided balloon dilation for benign anastomotic stricture after Ivor-Lewis esophagectomy: experience in 62 patients. AB - PURPOSE: To evaluate the safety and clinical effectiveness of fluoroscopically guided balloon dilation in 62 patients with benign anastomotic stricture after Ivor-Lewis esophagectomy. MATERIALS AND METHODS: Between January 1996 and June 2004, fluoroscopically guided balloon dilation was undertaken in 62 patients with benign anastomotic stricture after Ivor-Lewis esophagectomy. Radiologic images and medical records including complications were retrospectively reviewed. The maximum diameters of the balloon catheters used were 18-20 mm. Clinical success was defined by the absence of recurrent dysphagia after balloon dilation until the most recent follow-up. The Fisher exact test was used to assess the relationship of symptomatic recurrence and the balloon size, width of the stricture, and radiation therapy. RESULTS: There were 115 sessions of balloon dilation in 62 patients (mean, 1.85 sessions per patient). Clinical success was achieved in 59 patients (95%) by means of a single dilation (n = 29) or by multiple dilations (n = 30). One patient with severe stenosis was successfully treated with temporary placement of a covered retrievable stent. Major complications such as esophageal perforation or massive bleeding did not occur. Four patients with mucosal tear (n = 3) or aspiration pneumonia (n = 1) were conservatively treated. Patients with severe stricture had more symptomatic recurrences than those with moderate stricture. CONCLUSION: Fluoroscopically guided balloon dilation is a safe and successful treatment modality for benign anastomotic stricture after Ivor-Lewis esophagectomy. PMID- 16371539 TI - Fluoroscopically guided balloon dilation for patients with esophageal stricture after radiation treatment. AB - PURPOSE: To evaluate the safety and clinical effectiveness of fluoroscopically guided balloon dilation in patients with esophageal stricture after radiation therapy (RT). MATERIALS AND METHODS: From April 1993 through December 2004, fluoroscopically guided balloon dilation was performed in 15 patients with esophageal strictures secondary to previous RT. Technical success, clinical success, recurrence of dysphagia, primary and secondary patency rates, and complications related to the procedure were retrospectively evaluated. RESULTS: Twenty-five balloon dilations were performed in 15 patients, with a mean of 1.7 dilations per patient (range, 1-5). Technical success was achieved in all procedures. One patient was immediately lost to follow-up and another underwent elective esophageal surgery 13 days after the procedure. Of the remaining 13 patients, clinical success was achieved 11 (85%). Two of 13 patients exhibited recurrence of dysphagia before 1 month after balloon dilation. Among the 11 patients in whom clinical success was achieved, seven exhibited maintained initial improvement of dysphagia until their last follow-up (mean, 174 days) and four exhibited recurrence of dysphagia after the first balloon dilation. Dysphagia recurred 2-128 days (mean, 67.2 d) after the first balloon dilation in six of the 13 patients (46%), who underwent further balloon dilation and/or stent placement. The primary and secondary patency rates at 1, 3, and 6 months were 86%, 68%, and 47% and 100%, 92%, and 62%, respectively. There were no major complications. Type 1 and 2 esophageal ruptures occurred after 12 dilations in nine patients; they were treated conservatively. CONCLUSION: Fluoroscopically guided balloon dilation for esophageal stricture after RT can be safe and effective. However, the high rate of recurrent dysphagia requires repeated dilations. PMID- 16371540 TI - Effects of the type of embolization particles on carboplatin concentration in liver tumors after transcatheter arterial chemoembolization in a rabbit model of liver cancer. AB - PURPOSE: To study the effect of particle type used during transarterial hepatic chemoembolization (TACE) on carboplatin concentration after TACE in an animal model of liver cancer (VX2) and to determine the concentration of carboplatin within tumor, liver, and plasma. MATERIALS AND METHODS: The VX2 tumors were grown in the livers of 23 rabbits. Carboplatin (5 mg/kg) was selected because of its known potency against VX2 tumor. Group 1 was treated with TACE with tris-acryl gelatin microspheres (100-300 microm), group 2 was treated with TACE with polyvinyl alcohol (PVA; 150-250 microm), group 3 (control) was treated with intraarterial saline solution, and group 4 (pharmacokinetic) was treated with intraarterial carboplatin. Animals were killed after 48 hours, and concentrations of carboplatin were measured by atomic absorption spectroscopy from samples of blood and liver (central and peripheral zones of tumor and nontumorous liver tissue). RESULTS: In group 1 (tris-acryl gelatin microspheres) and group 2 (PVA), the mean carboplatin concentrations were 117 microg/g and 31.8 microg/g, respectively, within the central zone of the tumor and 38.5 mug/g versus 7.9 microg/g, respectively, in the peripheral zone. No carboplatin was detected in nontumorous liver tissue and plasma concentrations were low in both treated groups (<0.079 microg/mL). CONCLUSIONS: Carboplatin concentration was significantly greater (by a factor of two to four) within the central zone of the tumor compared with the peripheral zone in both treated groups. The overall tumor carboplatin concentrations were significantly greater in the tris-acryl gelatin microsphere group than in the PVA group (P < .001), which could translate into greater potency and tumor kill. Administration of tris-acryl gelatin microspheres may be clinically advantageous during TACE, as it contributed to greater delivery of chemotherapy to tumor in the present study. PMID- 16371541 TI - Time-related changes in computed tomographic appearance and pathologic findings after radiofrequency ablation of the rabbit lung: preliminary experimental study. AB - PURPOSE: To evaluate correlations between computed tomographic (CT) appearance and pathologic findings after radiofrequency (RF) ablation of lungs and to determine whether CT appearance could predict the extent of the effective therapeutic area. MATERIALS AND METHODS: The lungs of 14 rabbits were subjected to RF ablation and CT scans were obtained immediately and at various intervals thereafter. Four rabbits were killed immediately after the initial CT imaging (n = 4). The remaining 10 rabbits were killed after additional CT scans at intervals of 3 days (n = 2), 1 week (n = 4), 2 weeks (n = 2), and 3 weeks (n = 2) after RF ablation. Pathologic findings were correlated with CT appearance. RESULTS: Immediately after RF ablation, a restricted area of central dense opacity enclosed by an extensive area of ground-glass opacity was noted in the ablated region on CT images. Pathologically, the former corresponded to destructive tissue and the latter corresponded to tissue with some degree of injury. After 1 week, the entire ablated region appeared as a well-demarcated homogeneous dense opacity on CT that corresponded to necrotic tissue and its surrounding rim of granulation tissue on histopathologic examination, indicating that the enclosing extensive area of ground-glass opacity on the initial CT scan represented an ongoing necrosis. Within 2-3 weeks, the ablated region gradually contracted on the CT images, representing a tissue repairing process in which the granulation tissue was encroaching on the inner necrotic tissue. CONCLUSION: Ground-glass opacity of the ablated region on CT immediately after RF ablation represents an ongoing necrosis. PMID- 16371542 TI - Multiple-electrode radiofrequency ablation: simultaneous production of separate zones of coagulation in an in vivo porcine liver model. AB - PURPOSE: A multiple-electrode radiofrequency (RF) system was developed based on switching between electrodes that allows for the simultaneous use of as many as three electrically independent electrodes. The purpose of this study was to determine if each multiple-electrode ablation zone is identical to an ablation zone created with conventional single-electrode mode. MATERIALS AND METHODS: Nine female domestic pigs (mean weight, 90 kg) were used for this study. A prototype monopolar multiple-electrode RF ablation system was created with use of an RF generator and an electronic switching algorithm. A maximum of three electrodes can be used simultaneously by switching between electrodes at each impedance spike (30 omega greater than baseline levels). A total of 39 zones of ablation were created at open laparotomy in pig livers with use of a conventional single electrode (n = 9), two single electrodes simultaneously (n = 6 ablations; 12 ablation zones), or three single electrodes simultaneously (n = 6 ablations; 18 ablation zones). RF electrodes were spaced in separate lobes of the liver when multiple zones of coagulation were created simultaneously. Animals were euthanized after RF ablation, livers were removed, and ablation zones were sectioned and measured. RESULTS: Zones of coagulation created simultaneously with two or three electrodes were equivalent to ablation zones created with use of conventional single-electrode ablation. No significant differences were observed among control animals treated with a single electrode, those with two separate zones of ablation created simultaneously, and those with three simultaneously created ablation zones in terms of mean (+/-SD) minimum diameter (1.6 cm +/- 0.6, 1.6 cm +/- 0.5, and 1.7 cm +/- 0.4, respectively), maximum diameter (2.0 cm +/- 0.5, 2.3 cm +/- 0.5, 2.2 cm +/- 0.5, respectively), and volume (6.7 cm3 +/- 3.7, 7.4 cm3 +/- 3.8, and 7.8 cm3 +/- 3.9; P > .30, analysis of variance, pairwise t test comparisons). CONCLUSIONS: A rapid-switching multiple-electrode RF system was able to simultaneously create as many as three separate ablation zones of equivalent size compared with single-electrode controls. This system would allow physicians to simultaneously treat multiple tumors, substantially reducing procedure time and anesthesia risk. PMID- 16371543 TI - Acute aortoiliac and femoral artery thrombosis complicating diabetic ketoacidosis. AB - Acute abdominal aortic occlusion is a devastating event with high associated rates of morbidity and mortality even with surgical intervention. This report describes a case of acute aortoiliac and femoral artery occlusion likely resulting from a hypercoagulable state caused by diabetic ketoacidosis (DKA). Vascular thrombosis is a little-known but potentially devastating complication of DKA that should be considered in every patient treated for DKA and should be added to the differential diagnoses when attempting to determine the etiology of a thrombosed vessel. PMID- 16371544 TI - Chemoembolization of the left inferior phrenic artery in patients with hepatocellular carcinoma: radiographic findings and clinical outcome. AB - The purpose of this study was to evaluate the radiographic findings of hepatocellular carcinoma (HCC) supplied by the left inferior phrenic artery (LIPA) and the safety and efficacy of transcatheter arterial chemoembolization (TACE) of the LIPA. From September 2002 until August 2004, 11 patients with LIPA supplying HCC were identified and successfully treated with TACE of all LIPAs. LIPA collateral vessels may supply HCC, particularly when the tumors are in the left lobe of the liver in patients who have undergone previous TACE procedures. In this setting, such collateral vessels should be sought on preprocedural imaging. These LIPA collateral vessels may be safely embolized with very good clinical results. PMID- 16371545 TI - Prospective study of arterial infusion chemotherapy followed by radiofrequency ablation for the treatment of liver metastasis of gastric cancer. AB - This prospective study was conducted to evaluate the efficacy of hepatic arterial infusion chemotherapy (HAIC) with use of an implanted port followed by radiofrequency (RF) ablation for the treatment of liver metastasis of gastric cancer. Seven patients without extrahepatic metastasis were enrolled. The maximum tumor size was less than 3 cm in one patient and 3.2-6.0 cm in the other six patients (mean, 4.4 cm +/- 1.5). Two patients had single lesions and the other five patients had two to six lesions each. The maximum tumor size was reduced to 3 cm or less (mean, 2.4 cm +/- 0.4; P < .03) after HAIC in all patients. The total number of tumors was reduced from 25 to 16. RF ablation was performed for all residual liver tumors, resulting in complete tumor necrosis. All but one patient are still living without intrahepatic recurrence, with a median survival time of 16.5 months. PMID- 16371546 TI - Clinical services provided by interventional radiologists to Medicare beneficiaries in the United States, 2000-2003. AB - To identify trends in Evaluation and Management (E&M) and non-E&M services of interventional radiologists (physician specialty type 94) from 2000 to 2003 for Medicare patients, Medicare Part B physician annual allowed services data from the Centers of Medicare and Medicaid Services (CMS) were analyzed for all interventional radiologists from 2000 to 2003. Because the number of interventional radiologists in the United States according to the Society of Interventional Radiology is, on average, 4.2 times the number of interventional radiologists who use physician specialty type 94, we extrapolated the E&M services for each year. During the period examined, the total number of E&M services by interventional radiologists increased 309%, from 9,698 in 2000 to 29,914 in 2003. The most commonly performed services were Office or Other Outpatient Visit (Current Procedural Terminology [CPT] codes 99211-99215) for established patients, followed by Subsequent Hospital Care (CPT 99231-99233) and Office or Other Outpatient Consultations (CPT 99241-99245). The extrapolated number of E&M services by interventional radiologists for Medicare patients in 2003 is approximately 107,853. The number of Office and Outpatient Visits for New Patients (CPT 99201-99205) increased 142%, whereas the number of Consultations for New Patients (CPT 99241-99245) increased 208%. The total number of codes reimbursed by CMS to interventional radiologists (type 94) increased from 2.8 million in 2000 to 3.8 million in 2003. PMID- 16371547 TI - Repair of iatrogenic subclavian artery pseudoaneurysm with covered stent placement after vertebral artery transposition. PMID- 16371548 TI - Successful retrieval of a Gunther tulip vena cava filter with the assistance of a curved sheath introducer. PMID- 16371549 TI - Sonographically guided percutaneous muscle biopsy in diagnosis of neuromuscular disease: a useful alternative to open surgical biopsy. AB - OBJECTIVE: The purpose of this study was to evaluate the feasibility of sonographically guided percutaneous muscle biopsy in the investigation of neuromuscular disorders. METHODS: Sonographically guided percutaneous needle biopsy of skeletal muscle was performed with a 14-gauge core biopsy system in 40 patients over a 24-month period. Patients were referred from the Department of Neurology under investigation for neuromuscular disorders. Sonography was used to find suitable tissue and to avoid major vascular structures. A local anesthetic was applied below skin only. A 3- to 4-mm incision was made. Three 14-gauge samples were obtained from each patient. All samples were placed on saline dampened gauze and sent for neuropathologic analysis. As a control, we retrospectively assessed results of the 40 most recent muscle samples acquired via open surgical biopsy. RESULTS: With the use of sonography, 32 (80%) of 40 patients had a histologic diagnosis made via percutaneous needle biopsy. This included 26 (93%) of 28 patients with acute muscular disease and 6 (50%) of 12 patients with chronic disease. In the surgical group (all acute disease), 38 (95%) of 40 patients had diagnostic tissue attained. CONCLUSIONS: Sonographically guided percutaneous 14-gauge core skeletal muscle biopsy is a useful procedure, facilitating diagnosis in acute muscular disease. It provides results comparable with those of open surgical biopsy in acute muscular disease. It may also be used in chronic muscular disease but repeated or open biopsy may be needed. PMID- 16371550 TI - Gray scale contrast enhancement and quantification in different positions of rabbit liver. AB - OBJECTIVE: The purpose of this study was to evaluate the difference between central and ventral peripheral positions and the difference between left and right lobes in rabbit liver with gray scale contrast enhancement. METHODS: An in vivo model of perfusion was studied with a sulfur hexafluoride contrast agent and low-mechanical-index, real-time, gray scale harmonic imaging. The contrast agent (0.1 mL/kg body weight) was applied respectively in 10 rabbits by intravenous bolus injection. The time-intensity curve was used to obtain flow-related parameters such as time to enhancement (ET), time to peak intensity (PIT), peak signal intensity (PSI), enhancement duration (ED), and area under the curve (AUC). RESULTS: There was a significant difference in parameters of the time intensity curve between central and peripheral ventral positions of liver parenchyma (P < .05), except for the ED in the left liver. The ET and PIT were earlier, the PSI higher, the ED longer, and the AUC larger in the central position of parenchyma than in the peripheral position. In addition, the ET and PIT were earlier, the PSI higher, the ED longer, and the AUC larger in the right lobe of liver parenchyma than in the left lobe. There was a significant difference in parameters of the time-intensity curve between the left and right lobes of liver parenchyma (P < .05), except for the ET of the peripheral position. CONCLUSIONS: Flow parameters are different between central and ventral peripheral positions and between left and right lobes of hepatic parenchyma. PMID- 16371551 TI - In vivo perfusion estimation using subharmonic contrast microbubble signals. AB - OBJECTIVE: The purpose of this study was to quantify perfusion in vivo using contrast-enhanced subharmonic imaging (SHI). METHODS: A modified LOGIQ 9 scanner (GE Healthcare, Milwaukee, WI) operating in gray scale SHI mode was used to measure SHI time-intensity curves in vivo. Four dogs received intravenous contrast bolus injections (dose, 0.1 mL/kg), and renal SHI was performed. After 3 contrast agent injections, a microvascular staining technique based on stable (nonradioactive) isotope-labeled microspheres (BioPhysics Assay Laboratory Inc, Worcester, MA) was used to quantify the degree of perfusion in 8 sections of each kidney. Low perfusion states were induced by ligating surgically exposed segmental renal arteries followed by contrast agent injections and microvascular staining. Digital clips were transferred to a personal computer, and SHI time intensity curves were acquired in each section using Image-Pro Plus software (Media Cybernetics, Silver Spring, MD). Subharmonic fractional blood volumes were calculated, and the perfusion was estimated from the initial slope of the fractional blood volume uptake averaged over 3 injections. Subharmonic perfusion data were compared with the gold standard (ie, the microspheres) using linear regression analysis. RESULTS: In vivo gray scale SHI clearly showed flow and, thus, perfusion in the kidneys with almost complete suppression of tissue signals. In total, 270 SHI time-intensity curves were acquired, which reduced to 94 perfusion estimates after averaging. Subharmonic perfusion estimates correlated significantly with microsphere results (r = 0.57; P < .0001). The best SHI perfusion estimates occurred for high perfusion states in the anterior of the kidneys (r = 0.73; P = .0001). The corresponding root mean square error was 2.4%. CONCLUSIONS: Subharmonic perfusion estimates have been obtained in vivo. The perfusion estimates were in reasonable to good agreement with a microvascular staining technique. PMID- 16371552 TI - Imaging of peripheral cholangiocarcinoma with low-mechanical index contrast enhanced sonography and SonoVue: initial experience. AB - OBJECTIVE: The purpose of this study was to investigate the imaging findings of peripheral cholangiocarcinoma with low-mechanical index (MI) contrast-enhanced sonography. METHODS: Eighteen nodules of peripheral cholangiocarcinoma proved by pathologic examination in 18 patients were evaluated with contrast-enhanced sonography. A low-MI real-time contrast-enhanced sonographic mode (ie, contrast pulse sequencing) and a sulfur hexafluoride-filled microbubble contrast agent (SonoVue [BR1]; Bracco SpA, Milan, Italy) were used. RESULTS: On contrast enhanced sonographic images, all 18 nodules (100%) of peripheral cholangiocarcinoma showed inhomogeneous enhancement during the arterial phase, and the emergence of nodule enhancement was earlier in 3 nodules (16.7%), simultaneous in 13 (72.2%), and later in 2 (11.1%), respectively, when compared with the adjacent liver tissue. During the arterial phase, 8 nodules (44.4%) showed irregular peripheral rimlike hyperenhancement, 2 (11.1%) showed inhomogeneous hyperenhancement, and 8 (44.4%) showed inhomogeneous hypoenhancement. In portal and late phases, all 18 nodules (100%) showed hypoenhancement. When contrast-enhanced sonography was added for analysis, the confidence levels of the investigators were improved in 15 (83.3%) of 18 nodules, and 17 (94.4%) of 18 peripheral cholangiocarcinomas were correctly characterized. CONCLUSIONS: The imaging findings of peripheral cholangiocarcinoma had some characteristics on low-MI contrast-enhanced sonography. Knowledge of these characteristics might be beneficial for improving the diagnostic performance of sonography in evaluating this entity. PMID- 16371553 TI - Contrast-enhanced sonography of the lung for differential diagnosis of atelectasis. AB - OBJECTIVE: Because of the absence of air in atelectatic tissue, sonography allows visualization of lung atelectasis and may characterize pulmonary and bronchial arterial vascularity by contrast-enhanced sonography (CES). METHODS: Thirty consecutive patients with obstructive atelectasis (OA) (n = 17) and compression atelectasis (n = 13) were retrospectively studied by CES using a second generation sulfur hexafluoride contrast agent (SonoVue [BR1]; Bracco SpA, Milan, Italy). The following CES parameters were evaluated: (1) time to enhancement (TE) of the contrast agent after intravenous application was determined and classified as short TE and delayed TE (short TE, < or =6 seconds; versus delayed TE, >7 seconds); and (2) extent of enhancement (EE) was evaluated during the arterial phase (2-30 seconds) and the parenchymal phase (1-5 minutes): the EE of pleural lesions was determined in comparison with splenic enhancement and classified in reduced EE versus marked EE. RESULTS: All 13 patients with compression atelectasis had a short TE and a marked EE during arterial and parenchymal phases. In the remaining 17 patients with OA, 10 patients had a short TE and 7 patients had a delayed TE. The EE during both phases was reduced in 5 patients and marked in 3. Nine of 17 patients with OA had different EE during arterial and parenchymal phases. CONCLUSIONS: Compression atelectasis is characterized by CES with a short TE and a marked EE, indicating patent pulmonary arterial vascularization. In patients with OA, a variable CES pattern is found. With regard to only the TE, a delayed TE implies OA. This indicates a shifting of pulmonary vascularization to bronchial arterial vascularization in these patients. PMID- 16371554 TI - Left hemidiaphragmatic mobility: assessment with ultrasonographic measurement of the craniocaudal displacement of the splenic hilum and the inferior pole of the spleen. AB - OBJECTIVE: The purpose of this study was to evaluate the correlation between the radiographic and ultrasonographic measurements of craniocaudal displacement of the left hemidiaphragm. METHODS: Forty-nine patients with clinical indications for interventional procedures prospectively underwent radiographic evaluation of left hemidiaphragmatic mobility and B-mode ultrasonographic measurement of craniocaudal displacement of the hilum and the inferior pole of the spleen. Ultrasonography was performed with a 3.5-MHz convex transducer in a left intercostal position under a longitudinal orientation. Statistical analyses were performed with linear regression, a paired Student t test, and Bland-Altman analyses. RESULTS: The correlation between the craniocaudal splenic hilum displacement and radiographic measurements was found to be linear: hemidiaphragmatic mobility = 17.795 + 0.429 x splenic hilum displacement (SE for the regression coefficient = 0 .12; P = .0012), although the values obtained with both methods were statistically different (P < .05). The same results could be observed with the use of the inferior pole of the spleen: hemidiaphragmatic mobility = 9.5596 + 0.5455 x inferior polo displacement (SE for the regression coefficient = 0 .11; P < .0001). The mean difference between the values obtained by ultrasonography and by radiography was statistically significant (16.7 +/- 16.1 mm; P < .05 [hilum]; 18.9 +/- 14.2 mm; P < .05 [inferior pole]). CONCLUSIONS: These results allow us to conclude that ultrasonography can be used as an alternative method for left hemidiaphragm mobility evaluation compared with radiography. PMID- 16371555 TI - Feasibility of an infraclavicular block with a reduced volume of lidocaine with sonographic guidance. AB - OBJECTIVE: A successful brachial plexus block requires a large volume of a local anesthetic. Sonography allows reliable deposition of the anesthetic around the cords of the brachial plexus, potentially lowering the anesthetic requirement. METHODS: Fifteen sonographically guided infraclavicular blocks were performed in 14 patients with 2% carbonated lidocaine with epinephrine through a 17-gauge Tuohy needle. The amount of lidocaine injected at several points around each cord was based on satisfactory spread observed sonographically. A 19-gauge catheter was then placed with its tip between the posterior cord and axillary artery, and tip position was confirmed by observing the spread of 1 to 2 mL of injected air. Lidocaine was injected through the catheter if necessary to prolong the blocks. RESULTS: Surgery was performed in all patients without general anesthesia, rescue blocks, or infiltration. A heroin user was given an additional 50 microg of fentanyl before the block. One patient required 5 mL of lidocaine through the catheter for an incomplete radial nerve block 5 minutes after initial injection. Seven patients received additional midazolam (mean, 2.5 mg) for alleviation of anxiety despite excellent blocks. The mean +/- SD volume of lidocaine for the initial block was 16.1 +/- 1.9 mL (4.2 +/- 0.9 mg/kg). In 4 patients, additional lidocaine 1 hour after an initial successful block increased the total volume to 19.5 +/- 7.1 mL (5 +/- 1.9 mg/kg). The mean times to perform the block, onset of the block, and achieving surgical anesthesia and the duration of surgery were 10.8 +/- 3.3, 2 +/- 1.3, 5.9 +/- 2.6, and 92.7 +/- 54.4 minutes, respectively. CONCLUSIONS: A successful infraclavicular block in adults with 14 mL of lidocaine is feasible with the use of sonography. The reduced volume does not seem to affect the onset but shortens the duration of the block. PMID- 16371556 TI - Endosonographic imaging of anorectal diseases. AB - OBJECTIVE: The normal sonographic anatomy of the anorectum, sonographic findings of anorectal diseases, and indications and limitations of endosonography compared with magnetic resonance imaging are reviewed. Methods. Endosonographic imaging was performed with a Siemens (Erlangen, Germany) FI 400 ultrasound scanner with an end-fire 7.5-MHz biplane endorectal probe and a B-K Medical (Sandhoften, Denmark) scanner with an 1850 axial-type side-fire 5.0- to 10.0-MHz rotating endoscopic probe. RESULTS: Rectal carcinoma appears on endorectal sonography as a low-echogenicity lesion that abruptly interrupts the normal sequence of layers. The internal anal sphincter is seen very clearly on endoanal sonography, and it is easy to appreciate atrophy and small tears of this sphincter. Endoanal sonography cannot accurately show thinning of the external anal sphincter. Peroxide-enhanced endoanal sonography is especially useful for patients with recurrent perianal fistulas in whom scarring should be distinguished from recurrent fistulas and detection of the internal opening. However, sonography does not provide an adequate deep and global display of all adjacent pelvic and perineal spaces. CONCLUSIONS: Endosonography can accurately stage primary rectal tumors and assess the internal anal sphincter. Peroxide-enhanced 3-dimensional imaging can increase the utility of endoanal sonography in detection and characterization of perianal fistulas and planning of optimal therapy. However, magnetic resonance imaging can be used a complementary modality to endosonography, especially for evaluation of external anal sphincter atrophy and deep pelvic inflammation. PMID- 16371557 TI - High-resolution sonographic spectrum of diverticulosis, diverticulitis, and their complications. AB - OBJECTIVE: The purpose of this study was to evaluate the high-resolution sonographic features of diverticulosis, diverticulitis, and their complications. METHODS: During a period of about 4 years 8 months, there were 25 patients with sonographic features of diverticulosis, uncomplicated diverticulitis, and complicated diverticulitis. The clinical symptoms, sonographic features, follow up investigations, and management details were recorded. RESULTS: The common symptoms were pain in the left lower quadrant and fever. Sonographic features of uncomplicated diverticulitis were a varying appearance of the diverticulum with pericolic inflammation. Colonic wall thickening was not a consistent sign. Complications seen were pericolic, mesocolic, and intraperitoneal abscesses, colovesical fistulas, colouterine fistulas, perforation, and small-bowel obstruction. Uninflamed diverticula were seen in all patients with left-sided disease. They had 7 types of sonographic appearances. CONCLUSIONS: Uncomplicated diverticulitis is seen as a diverticulum of variable echogenicity with pericolic inflammation. An inflamed diverticulum is not visualized in complicated diverticulitis. Visualization of uninflamed diverticula helps reinforce the diagnosis of uncomplicated diverticulitis and predict the cause in complicated diverticulitis. PMID- 16371558 TI - Sonography of the gastrointestinal tract. AB - OBJECTIVE: The purpose of this presentation is to provide an overview of sonographic manifestations of various gastrointestinal diseases. METHODS: Patients were scanned with an HDI 3000 or HDI 5000 system (Philips Medical Systems, Bothell, WA) with both a 2- to 5-MHz curvilinear transducer and a 5- to 12-MHz linear transducer. The patients were asked to fast from the previous night. For accurate visualization of the stomach, a cup of water was given to patients thought to have stomach or upper gastrointestinal disease. RESULTS: A satisfactory sonographic examination could be performed in all patients. CONCLUSIONS: With recent technical advances, the increasing use of sonography in initial evaluation of patients with abdominal pain may allow earlier detection of bowel disease. PMID- 16371559 TI - Unusual presentations of midgut volvulus with the whirlpool sign. AB - OBJECTIVE: The purpose of this series was to emphasize the varied presentations of midgut volvulus from neonatal life to well into adulthood and to evaluate the role of the whirlpool sign on sonography in diagnosing this condition. METHODS: A retrospective search of all midgut volvulus cases diagnosed in the radiology department of a 1500-bed general hospital between February 2002 and July 2005 was done. Eight patients with surgically confirmed midgut volvulus were found, all of whom had undergone sonography as the initial screening investigation. RESULTS: Eight patients, with ages ranging from 9 days to 32 years (mean age, 11.8 years), had undergone sonography for varied conditions, including bilious and nonbilious vomiting, vague abdominal pain, epigastric lumps, weight loss, features mimicking appendicitis, and pancreatitis. In all patients, the diagnosis of midgut volvulus was made on the basis of the clockwise whirlpool sign on sonography. Subsequently, 4 patients underwent upper gastrointestinal series, and contrast enhanced computed tomography was done in 6 cases. Surgical confirmation of the diagnosis was obtained in all cases. Two patients were taken for surgery on the basis of only sonographic findings. CONCLUSIONS: Midgut volvulus can also appear much beyond the neonatal age group with a variety of clinical presentations, making the clinical diagnosis in such patients very difficult, if not impossible. Identification of the clockwise whirlpool sign on sonography is an accurate way of diagnosing this condition, which can preclude the need for further investigations and can allow prompt surgical intervention. PMID- 16371560 TI - Sonographic features of internal hernia. AB - OBJECTIVE: The purpose of this series is to describe the sonographic findings in 4 patients with intestinal obstruction due to internal hernia. METHODS: Four patients had clinical features of intestinal obstruction. Sonography was performed with broadband convex and linear array transducers in these patients. RESULTS: In all 4 patients, there were features of intestinal obstruction with a zone of transition between dilated and nondilated bowel. There was a cluster of crowded, compressed, and aperistaltic small-bowel loops, as if they were tightly packed within a sac, by the side of this zone in 3 patients. This appearance was due to obstruction of the afferent loop caused by compression. In the last patient, there were 2 loops of dilated bowel within a sac outlined by fluid due to obstruction of the efferent loop. All 4 patients had obstructed internal hernias at laparotomy. CONCLUSIONS: Sonographic features of internal hernia are described. PMID- 16371561 TI - Spongioid hyperplasia of the gallbladder. PMID- 16371562 TI - Prenatal sonographic diagnosis of Grebe syndrome. PMID- 16371563 TI - Cyst of the canal of Nuck with demonstration of the proximal canal: the role of the compression technique in sonographic diagnosis. PMID- 16371564 TI - Unilateral hyperechoic renal pyramids in a child with renal artery stenosis. PMID- 16371565 TI - Contrast-enhanced low-mechanical-index ultrasonography in hepatic splenosis. PMID- 16371566 TI - Sonographic detection of synchronous occurrence of benign nodules in orthotopic and ectopic thyroid tissue. PMID- 16371567 TI - Extratesticular scrotal calcifications: their relationship with sports. PMID- 16371568 TI - In vivo destruction of ultrasound contrast microbubbles is independent of the mechanical index. PMID- 16371569 TI - Mother and adolescent representations of illness ownership and stressful events surrounding diabetes. AB - OBJECTIVE: To assess the extent to which adolescents with diabetes and their mothers appraise diabetes as a shared entity across adolescence through (a) assessing appraisals of illness ownership and their relationship to joint responsibility for daily diabetes tasks, (b) exploring whether appraisals of shared illness ownership are associated with congruent views of what is stressful about diabetes, and (c) examining whether age-related declines occur in these shared appraisals across adolescence. METHODS: One hundred twenty-seven adolescents (ages 10-15 years, M = 12.8) and their mothers completed an interview that probed appraisals of illness ownership, the most stressful events surrounding diabetes in the past week, and a questionnaire regarding who was responsible for performing diabetes-related tasks. RESULTS: Dyads, most frequently, agreed that diabetes was a "shared" entity. Shared appraisals of illness ownership reflected the greater joint responsibility of mothers and children in daily diabetes tasks. Shared appraisals of illness ownership were not related to congruent reports of diabetes stressful events, and incongruence in appraisals of stressful events was common. With age adolescents reported less shared illness ownership and congruence regarding stressful events, age differences were not seen in mothers' reports. CONCLUSIONS: Although diabetes is often appraised as a social entity, adolescents and their mothers experience different aspects of the disease as stressful, especially as adolescents age, and become more independent in performing diabetes-related tasks. PMID- 16371570 TI - Commentary: the role of memory in managing children's distress during medical procedures. PMID- 16371571 TI - Weight preoccupation as a function of observed physical attractiveness: ethnic differences among normal-weight adolescent females. AB - OBJECTIVE: To examine the association between observer ratings of physical attractiveness and weight preoccupation for female adolescents, and to explore any ethnic differences between Caucasian, African-American, and Hispanic females. METHODS: Normal-weight female adolescents who had participated in the National Longitudinal Study of Adolescent Health in-home Wave II survey were included (n = 4,324). Physical attractiveness ratings were made in vivo by interviewers. Using logistic regression models stratified by ethnicity, the associations between observer-rated attractiveness and weight preoccupation were examined after controlling for demographics, measured body mass index (BMI) and psychosocial factors. RESULTS: Caucasian female adolescents perceived as being more attractive reported significantly greater weight preoccupation compared with those rated as being less attractive. Observed attractiveness did not relate to weight preoccupation among African-American or Hispanic youth when controlling for other factors. CONCLUSIONS: For Caucasian female adolescents, being perceived by others as more attractive may be a risk factor for disordered eating. PMID- 16371572 TI - Perinatal olfactory learning in the domestic dog. AB - The ability of individuals to learn about chemosensory stimuli in the prenatal, or immediate postnatal, period may be advantageous in acquiring information about "safe" foods after weaning. In this study, we examined the influence of perinatal exposure to aniseed via the mother's diet on a two-choice food test in the domestic dog. Pups were tested at 10 weeks of age following "prenatal" exposure to aniseed (the last 20 days of gestation), "postnatal" exposure to aniseed (the first 20 days after birth), "perinatal" exposure to aniseed (pre- and postnatal exposure combined), or no exposure to aniseed prenatally or postnatally (control). Perinatal exposure resulted in a significantly greater preference for the aniseed food than the other types of exposure. At 10 weeks, there was no evidence for the retention of any prenatal learning of the aniseed. It is suggested that exposure to a chemosensory stimulus across the perinatal period results in a greater effect than simply the sum of pre- and postnatal exposure due to priming of the chemosensory system via prenatal chemosensory experience. Such a system may confer survival advantages by promoting the acquisition of information about safe foods. PMID- 16371573 TI - Pulse stimulation with odors or IBMX/forskolin potentiates responses in isolated olfactory neurons. AB - Many odor responses are mediated by the adenosine 3',5'-cyclic monophosphate (cAMP) pathway in which the cAMP-gated current is amplified by Ca2+-dependent Cl- current. In olfactory neurons, prolonged exposure to odors decreases the odor response and is an adaptive effect. Several studies suggest that odor adaptation is linked to elevated intracellular Ca2+. In the present study, using the perforated configuration of the patch clamp technique, we found that repetitive odor stimulation elicits a potentiation of the subsequent responses in olfactory neurons. This potentiation is mimicked by stimulating the cAMP pathway and does not appear to be related to phosphorylation of ion channels since protein kinase inhibitors could not block it. Our data suggest that local increases in [Ca2+]i via activation of the cAMP pathway mediate the pulse-elicited potentiation. In the first odor application, entry of Ca2+ through cyclic nucleotide-gated channels appears to be buffered. Repetitive stimulation allows local increases in [Ca2+]i, recruiting more Ca2+-dependent Cl- channels with each subsequent odor pulse. PMID- 16371574 TI - MR imaging in patients with Crohn disease: value of T2- versus T1-weighted gadolinium-enhanced MR sequences with use of an oral superparamagnetic contrast agent. AB - PURPOSE: To prospectively compare oral contrast-enhanced T2-weighted half-Fourier rapid acquisition with relaxation enhancement (RARE) magnetic resonance (MR) imaging with T1-weighted gadolinium-enhanced fast low-angle shot (FLASH) MR and standard examinations in the evaluation of Crohn disease. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. Fifty nine patients with Crohn disease underwent MR imaging after oral administration of a superparamagnetic contrast agent; RARE plain and fat-suppressed sequences and FLASH sequences were performed before and after intravenous injection of gadolinium chelate. References were endoscopic, small-bowel barium, computed tomographic, ultrasonographic, and clinical-biochemical scoring of disease activity. Two radiologists analyzed MR images for presence and extent of Crohn disease lesions, presence of strictures or other complications, and degree of local inflammation. MR findings were correlated with endoscopic, radiologic, and clinical data (kappa statistic and Spearman rank correlation test). RESULTS: T2 weighted MR was 95% accurate, 98% sensitive, and 78% specific for detection of ileal lesions. Agreement between T1- and T2-weighted images ranged from 0.77 for ileal lesions to 1.00 for colic lesions. T2-weighted MR enabled detection of 26 of 29 severe strictures, 17 of 24 enteroenteric fistulas, and all adhesions and abscesses; T1-weighted MR enabled detection of 20 of 29 severe strictures, 16 of 24 enteroenteric fistulas, and all adhesions and abscesses. Complications leading to surgery were found in 12 (20%) patients; these were assessed correctly with either T1- or T2-weighted images. T2-weighted signal intensities of the wall and mesentery correlated with biologic activity (P < .001, r of 0.774 and 0.712, respectively). Interobserver agreement was 0.642-1.00 for T2-weighted and 0.711 1.00 for T1-weighted images. CONCLUSION: T2-weighted MR can depict Crohn disease lesions and help assess mural and transmural inflammation with the same accuracy as gadolinium-enhanced T1-weighted MR. Combination of gadolinium-enhanced T1- and T2-weighted sequences is useful in the assessment of Crohn disease. PMID- 16371575 TI - Optimal peak systolic velocity threshold at duplex us for determining the need for carotid endarterectomy: a decision analytic approach. AB - PURPOSE: To determine the optimal peak systolic velocity (PSV) threshold at duplex ultrasonography (US) required to establish the need for carotid endarterectomy in symptomatic patients on the basis of the long-term cost effectiveness outcomes of diagnostic testing and subsequent treatment. MATERIALS AND METHODS: From January 1997 through January 2000, a prospective medical ethics committee-approved multicenter study was conducted. After giving informed consent, patients with amaurosis fugax, transient ischemic attack, or minor stroke who underwent duplex US and digital subtraction angiography were included in the study. Selective ipsilateral carotid angiograms were obtained in at least three planes. Arteries that were nearly or totally occluded at duplex US were excluded because the PSV cannot be reliably measured in these vessels. Receiver operating characteristic (ROC) curves were constructed for the diagnoses of 70% 99% and 50%-99% stenoses. Optimal likelihood ratios were calculated on the basis of lifetime costs and quality-adjusted life-years derived at cost-effectiveness analysis and the prevalence of disease. The associated optimal sensitivities, specificities, and PSV thresholds were derived from the ROC curves. RESULTS: In this clinical study, 350 patients were included. The nonoccluded arteries in a total of 236 patients were assessable for ROC analysis. For the diagnosis of 70% 99% stenosis, the optimal likelihood ratio was 0.21, which was associated with a PSV threshold of 220 cm/sec, a sensitivity of 97% (127 of 131 patients; 95% confidence interval [CI]: 94%, 100%), and a specificity of 48% (50 of 105 patients; 95% CI: 38%, 57%). For the diagnosis of 50%-99% stenosis, the optimal likelihood ratio was 0.38, which was associated with a PSV threshold of 180 cm/sec, a sensitivity of 95% (182 of 191 patients; 95% CI: 92%, 98%), and a specificity of 69% (31 of 45 patients; 95% CI: 55%, 82%). CONCLUSION: On the basis of the lifetime outcomes of diagnostic testing and subsequent treatment, the optimal PSV thresholds for the diagnosis of 70%-99% and 50%-99% carotid artery stenoses in patients with amaurosis fugax, transient ischemic attack, or minor stroke were 220 cm/sec and 180 cm/sec, respectively. PMID- 16371576 TI - Steroid myopathy: evaluation of fiber atrophy with T2 relaxation time--rabbit and human study. AB - PURPOSE: To determine whether muscle fiber atrophy associated with steroid myopathy can be detected with T2 relaxation time. MATERIALS AND METHODS: Animal and human studies were approved by the ethics committee. Informed consent was obtained. Twelve rabbits were divided into a group that received 3 mg/kg of triamcinolone subcutaneously each day for 10 consecutive days (n = 6) and a control group that received saline (n = 6). Magnetic resonance (MR) imaging was performed before and after treatment. T2 and fat deposition ratio (FDR) of soleus and gastrocnemius muscles before and after treatment and between control rabbits and rabbits treated with steroids were compared by using two-way repeated analysis of variance and Bonferroni post hoc test to evaluate effects of steroid treatment. After imaging, rabbits were sacrificed. Extracellular space ratio (ECSR) and fiber diameter were examined. Correlation among T2, ECSR, and diameter of type 2 muscle fibers was analyzed with a Pearson correlation test with Bonferroni correction in gastrocnemius to determine factors affecting T2. In humans, T2 relaxation time and FDR of both muscles were compared between volunteers not treated with steroids and patients treated with steroids by using an unpaired t test to evaluate the effects of steroids. RESULTS: In rabbits, T2 of gastrocnemius muscle was significantly (P < .01) longer after steroid treatment than before steroid treatment and was also significantly (P < .01) longer than after saline administration. T2 of the gastrocnemius showed no significant difference in control rabbits before or after saline administration or in control rabbits and rabbits before steroid administration. T2 of the soleus muscle or FDR of either muscle showed no significant difference. There was a significant correlation (P < .01) among T2, ECSR, and diameter of type 2 muscle fibers in the gastrocnemius. In humans, T2 of the gastrocnemius was significantly (P < .01) longer in patients than in volunteers. T2 of the soleus or FDR of either muscle showed no significant difference. CONCLUSION: Muscle fiber atrophy associated with steroid myopathy is detectable as prolongation of T2 relaxation time in the gastrocnemius muscle; the authors believe prolongation of T2 relaxation time is mainly due to increased ECSR reflecting type 2 muscle fiber atrophy. PMID- 16371577 TI - Hepatocellular carcinoma in patients with chronic liver disease: comparison of SPIO-enhanced MR imaging and 16-detector row CT. AB - PURPOSE: To compare the sensitivity, positive predictive value, and diagnostic accuracy of superparamagnetic iron oxide (SPIO)-enhanced magnetic resonance (MR) imaging with those of 16-detector row computed tomography (CT) for the detection of hepatocellular carcinoma (HCC) in patients with hepatitis B-induced cirrhosis. MATERIALS AND METHODS: Institutional Review Board approval was obtained for this study, and informed consent was obtained from all patients. A total of 44 patients (36 men, eight women; age range, 35-67 years) with 59 HCCs and mild liver cirrhosis (Child-Pugh score A or B) underwent multiphasic CT and SPIO enhanced MR imaging. The diagnosis of HCC was established at surgical resection (n = 31) and percutaneous biopsy (n = 28). SPIO-enhanced MR imaging was composed of T2-weighted turbo spin-echo and T2*-weighted gradient-echo sequences. Multiphasic CT consisted of four phases (ie, early arterial, late arterial, portal venous, and equilibrium). Three observers independently analyzed each image in random order. Sensitivity, positive predictive value, and diagnostic accuracy were calculated by using the alternative free-response receiver operating characteristic analysis for multi-detector row CT and SPIO-enhanced MR imaging. RESULTS: Although not significant (P > .05), the area under the receiver operating characteristic curve for SPIO-enhanced MR imaging (mean, 0.90) was higher than that for multi-detector row CT (mean, 0.82) for all observers. Also, although no significant difference was demonstrated by any of the three observers (P > .05), there was a trend toward increased sensitivity on both a per-lesion and a per-patient basis for SPIO-enhanced MR imaging (mean, 84.7% and 94.7%, respectively) compared with multi-detector row CT (mean, 76.9% and 88.6%, respectively). No significant difference in positive predictive value was observed between modalities. CONCLUSION: SPIO-enhanced MR imaging and multiphasic CT show similar diagnostic accuracy, sensitivity, and positive predictive value for the detection of HCC in patients with relatively mild hepatitis B-induced cirrhosis. PMID- 16371578 TI - Mitral regurgitation: quantification with 16-detector row CT--initial experience. AB - PURPOSE: To prospectively determine if retrospectively electrocardiographic (ECG) gated multi-detector row computed tomography (CT) with a 16-detector row CT scanner can depict mitral regurgitation and enable quantification of the severity of the disease. MATERIALS AND METHODS: The study had institutional review board approval, and patients gave informed consent. Nineteen patients with mitral regurgitation (10 men, nine women; mean age, 66 years +/- 9 [standard deviation]; range, 41-83 years) and 25 patients without mitral regurgitation (14 men, 11 women; mean age, 68 years +/- 9; range, 43-83 years) as determined with transesophageal color Doppler echocardiography and ventriculography underwent retrospectively ECG-gated 16-detector row CT. Twenty CT data sets covering the entire mitral valve apparatus were reconstructed in 5% steps of the R-R interval for each patient, and data analysis was performed with four-dimensional software. Using planimetry, two readers measured in consensus the area of the regurgitant orifice during systole. These measurements were compared with semiquantitative data from transesophageal echocardiography and ventriculography by using Spearman rank order correlation coefficients. RESULTS: In the 25 patients without mitral regurgitation, no regurgitant orifice during systole could be detected with multi detector row CT. In the 19 patients with mitral regurgitation, a regurgitant orifice could be visualized in all cases. The mean regurgitant orifice area at CT 45 mm(2) +/- 34 (range, 10-148 mm(2))-correlated significantly with the results at transesophageal echocardiography (r = 0.807, P < .001) and ventriculography (r = 0.922, P < .001). CONCLUSION: Planimetric measurements of the regurgitant orifice area at retrospectively ECG-gated 16-detector row CT enable quantification of mitral regurgitation. PMID- 16371579 TI - Simulation of liver lesions for pediatric CT. AB - PURPOSE: To develop and validate a technique based on characteristics of real lesions for simulating realistic small liver lesions on pediatric computed tomographic (CT) images. MATERIALS AND METHODS: The institutional review board provided exempt status for this study, determined that it was not subject to HIPAA compliance, and did not require informed consent. Patient identification information was removed from clinical images from contrast material-enhanced multi-detector row CT examinations performed in 10 children. Patients were infants or children up to 18 years old. Information about sex was not available. Children had one or more liver lesions of 2-6 mm in maximum transverse diameter. Images with more than one lesion were rendered multiple times, and each time, all but one of the lesions were digitally removed in sequence. This process provided images (n = 19) with a single real lesion. For consistency, the same image backgrounds (images with all real lesions removed) were used to create an identical number of images (n = 19), each with a single simulated lesion. Subsequently, three radiologists independently assessed images of real and simulated lesions that were presented in random order with a score on a continuous scale of 0 (definitely simulated) to 100 (definitely real). Mixed model analysis of variance was used to test the null hypothesis that the difference in population mean scores between the two lesion types was zero. RESULTS: The observer study did not reveal a significant difference in the ability of any radiologist to discriminate between real and simulated lesions (P > .31). The differences in mean scores for discrimination between real and simulated lesions for the three observers were -6, 9, and -7, respectively. The estimated overall difference was -1. CONCLUSION: Mathematic simulation of liver lesions is a feasible technique for creating realistic lesions for image quality or dose reduction studies in pediatric CT. PMID- 16371580 TI - Dutch iliac stent trial: long-term results in patients randomized for primary or selective stent placement. AB - PURPOSE: To determine long-term results of the prospective Dutch Iliac Stent Trial. MATERIALS AND METHODS: The study protocol was approved by local institutional review boards. All patients gave written informed consent. Two hundred seventy-nine patients (201 men, 78 women; mean age, 58 years) with iliac artery disease were randomly assigned to undergo primary stent placement (143 patients) or percutaneous transluminal angioplasty (PTA) with selective stent placement in cases in which the residual mean pressure gradient was greater than 10 mm Hg across the treated site (136 patients). Before and at 3, 12, and 24 months and 5-8 years after treatment, all patients underwent assessment, which included duplex ultrasonography (US), ankle-brachial index (ABI) measurement, Fontaine classification of symptoms, and completion of the Rand 36-Item Health survey for quality-of-life assessment. Treatment was considered successful for symptoms if symptoms increased at least one Fontaine grade, for ABI if ABI increased more than 0.10, for patency if peak systolic velocity ratio at duplex US was less than 2.5, and for quality of life if the RAND 36-Item Health Survey score increased more than 15 points. Effects of both treatments on symptoms, quality of life, patency, and ABI were compared by using survival analyses. RESULTS: Patients who underwent PTA and selective stent placement had better improvement of symptoms (hazard ratio [HR], 0.8; 95% confidence limits [CLs]: 0.6, 1.0) than did patients treated with primary stent placement, whereas ABI (HR, 0.9; 95% CLs: 0.7, 1.3), iliac patency (HR, 1.3; 95% CLs: 0.8, 2.1), and score for quality of life for nine survey dimensions did not support a difference between treatment groups. CONCLUSION: Patients treated with PTA and selective stent placement in the iliac artery had a better outcome for symptomatic success compared with patients treated with primary stent placement, whereas data about iliac patency, ABI, and quality of life did not support a difference between groups. PMID- 16371581 TI - Dynamic contrast-enhanced breast MR imaging in men: preliminary results. AB - PURPOSE: To prospectively evaluate whether the descriptors of lesion features and the diagnostic criteria that have been established for breast magnetic resonance (MR) imaging in female patients may be used for differential diagnosis with breast MR imaging in male patients as well. MATERIALS AND METHODS: The study design was approved by the institutional review board; all patients gave informed consent. The Institutional Review Board and informed consent information applied to the prospective and any retrospective component of the study. Seventeen consecutive male patients (mean age, 53 years +/- 14) were referred for imaging of a palpable breast mass. In addition to mammography and high-frequency breast ultrasonography, patients underwent dynamic breast MR imaging in a prone position with a dedicated double-breast surface coil. The standardized protocol consisted of a T2-weighted turbo spin-echo sequence followed by a dynamic series. Findings were recorded by using the terminology and descriptors and by evaluating the diagnostic criteria (related to morphology and enhancement kinetics) that have been developed for breast MR imaging in female patients. Validation was achieved at biopsy (nine patients) or follow-up with clinical examination and conventional imaging (eight patients). Because of the small size of the patient cohort, statistical significance was not tested. RESULTS: A total of 24 breast abnormalities were diagnosed. Three patients had invasive breast cancer (five tumors), 11 had gynecomastia (six unilateral, five bilateral), two had pseudogynecomastia, and one had a benign solid tumor (angiolipoma). All malignant tumors appeared as irregular masses with heterogeneous internal architecture or rim enhancement and showed rapid initial enhancement (mean value, 137% +/- 23) followed by a washout time course (Breast Imaging Reporting and Data System [BI RADS] category 5). Diffuse and nodular gynecomastia showed slow initial and persistent enhancement with normal-appearing parenchymal architecture (BI-RADS category 2; 15 of 16 breasts in 10 of 11 patients). In one patient with biopsy proved bilateral gynecomastia, an area with segmental enhancement was classified as suspicious for ductal carcinoma in situ. Pseudogynecomastia did not enhance at all. The angiolipoma showed benign morphologic features and slow initial and persistent enhancement (BI-RADS category 2). CONCLUSION: In the small study cohort, the MR imaging features of benign breast diseases and breast cancers in male patients seemed to be comparable to those seen in female patients. PMID- 16371582 TI - Distinguishing benign from malignant adrenal masses: multi-detector row CT protocol with 10-minute delay. AB - PURPOSE: To retrospectively evaluate the accuracy of precontrast attenuation, relative percentage washout (RPW), and absolute percentage washout (APW) in distinguishing benign from malignant adrenal masses at multi-detector row computed tomography (CT). MATERIALS AND METHODS: This HIPAA-compliant retrospective study had institutional review board approval; the need for informed consent was waived. One hundred twenty-two adrenal masses were evaluated in 99 patients (51 men, 48 women; age range, 37-86 years) who had undergone CT performed according to the study protocol and who either were given a pathologic diagnosis or underwent follow-up imaging. Unenhanced images were obtained before administration of 120 mL of an intravenous contrast agent with a 75-second scan delay. Delayed images were obtained after 10 minutes. RPW and APW were computed. Receiver operating characteristic (ROC) analysis was performed to compare mean attenuation and both RPW and APW. Analysis was first performed with the exclusion of pheochromocytomas, myelolipomas, and cysts. Precontrast attenuation criteria specific for benignity or malignancy were determined, and ROC analysis of results for the entire nonpheochromocytoma group was then performed. RESULTS: By using an RPW of 37.5% and excluding cysts and myelolipomas, all malignant lesions were detected with a sensitivity of 100% (17 of 17 lesions) and a specificity of 95% (90 of 95 lesions). Area under the binomial ROC curve (A(z)) values were 0.912, 0.985, and 0.892 for precontrast attenuation, RPW, and APW, respectively. Precontrast attenuation of less than 0 or more than 43 HU indicated benign and malignant entities, respectively. Incorporation of these criteria into the APW analysis yielded a sensitivity of 100% (17 of 17 lesions) and a specificity of 98% (93 of 95 lesions) for a threshold washout value of 52.0%. This attenuation corrected APW generated the greatest A(z) value (ie, 0.988). Combining all the information available from the protocol yielded a sensitivity of 100% (17 of 17 lesions) and a specificity of 98% (98 of 100 lesions) for differentiating benign from malignant masses. CONCLUSION: Precontrast attenuation of less than 0 HU supercedes the washout profile in the evaluation of an individual adrenal mass. Noncalcified, nonhemorrhagic adrenal lesions with precontrast attenuation of more than 43 HU should be considered suspicious for malignancy. PMID- 16371583 TI - Bone involvement in Erdheim-Chester disease: imaging findings including periostitis and partial epiphyseal involvement. AB - PURPOSE: To retrospectively review the bone findings at radiography, scintigraphy, computed tomography (CT), and magnetic resonance (MR) imaging in 11 patients with immunohistochemical and histologic proof of Erdheim-Chester disease. MATERIALS AND METHODS: This study was designed as a retrospective review; approval of the institutional review board and patient consent were not required for this type of study. Eleven patients (eight men and three women; mean age, 49 years; range, 17-68 years) with Erdheim-Chester disease underwent conventional radiography of the skeleton and bone scintigraphy. Two patients underwent CT of the femora and 10 underwent CT of the skull. Eight patients underwent MR imaging. Conventional radiographs, bone scintigrams, CT scans, and MR images were reviewed in consensus by four musculoskeletal radiologists. RESULTS: All 11 patients had involvement of the long bones and normal axial skeleton, hands, and feet. Bilateral and symmetric osteosclerosis of the diaphysis of the long bones was present in 52 (26 pairs) (98%) of the 53 bone lesions visible on conventional radiographs. Osteosclerosis was heterogeneous in 65% of the patients and homogeneous in 35%. Diaphysis was involved in 100% and metaphysis in 44 (83%) lesions. Partial epiphyseal involvement sparing the subchondral bone was present in 24 (45%) lesions. Periostitis was seen in 35 (66%) and endosteitis in 50 (94%) of the 53 long bones involved. Bone scintigraphy depicted tracer uptake in all bone lesions visible on radiographs. Skull and face bone lesions were present in two patients. MR imaging depicted a replacement of the normal fatty bone marrow by heterogeneous signal intensity on T1- and T2-weighted spin-echo images. Lesion extent, epiphyseal involvement, and periostitis were clearly depicted at MR imaging. CONCLUSION: This series provides a detailed description of bone involvement in Erdheim-Chester disease. Periostitis and partial epiphyseal involvement of the long bones are also features of this disease. (c) RSNA, 2005. PMID- 16371584 TI - Perfusion impairment in patients with normal-appearing coronary arteries: identification with contrast-enhanced MR imaging. AB - PURPOSE: To prospectively determine the feasibility of using first-pass magnetic resonance (MR) imaging to distinguish between myocardial segments in patients with coronary artery disease (CAD) of different degrees of obstruction and those in patients with normal-appearing coronary arteries. MATERIALS AND METHODS: The study was approved by the institutional ethics committee, and all patients provided informed consent. First-pass contrast material-enhanced MR imaging was performed at rest and after the infusion of dipyridamole in 37 patients (29 men, eight women; mean age, 57.2 years +/- 10.5 [standard deviation]) who had positive exercise test results or a clinical history of CAD. Myocardial segments were divided into five groups according to the degree of obstruction in the supplying artery. Signal intensity upslope, peak signal intensity, and time to peak signal intensity, as well as hyperemia-to-rest (HR) ratios for each of these three variables, were analyzed for each segment by using a generalized linear model. RESULTS: Signal intensity upslope in patients with normal coronary arteries at angiography was significantly higher than that in patients with CAD (P < .001). Signal intensity upslope for segments in patients without CAD was significantly different from that for normal-appearing segments in patients with CAD (P < .001). Signal intensity upslope (P < .05) and peak signal intensity (P < .01) enabled the differentiation of segments with more than 70% reduction in luminal diameter from those in all other groups. HR ratios demonstrated findings that were similar to those obtained by using each signal intensity variable alone. CONCLUSION: First-pass MR imaging can be used to distinguish segments with different degrees of obstructive CAD. Importantly, MR imaging can help identify segments with impaired perfusion and normal-appearing coronary arteries in patients with CAD and can demonstrate obstructive lesions in other territories. PMID- 16371585 TI - Pathologic skeletal muscle perfusion in patients with myositis: detection with quantitative contrast-enhanced US--initial results. AB - PURPOSE: To prospectively determine whether contrast material-enhanced ultrasonography (US) can depict inflammation-induced changes in muscle perfusion for patients suspected of having dermatomyositis or polymyositis and to compare these findings with those of magnetic resonance (MR) imaging and muscle biopsy. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. Perfusion in skeletal muscles was quantified with contrast enhanced intermittent power Doppler US by applying a modified model that analyzed the replenishment kinetics of microbubbles. In 22 patients (16 women, six men; mean age, 52 years +/- 17) who were suspected of having myositis and in 10 healthy volunteers (two women, eight men; mean age, 28 years +/- 4), contrast enhanced US of the clinically affected right biceps muscle was performed to measure blood flow, blood volume, and blood flow velocity. Additionally, the right upper arm was examined with a 1.5-T unit by using three different MR imaging techniques. Findings were compared with the results of clinical examinations and muscle biopsy. Data for perfusion-related parameters obtained at contrast-enhanced US were analyzed by using a nonparametric Mann-Whitney U test. RESULTS: Eight patients had histologically confirmed myositis and showed significantly higher blood flow velocity (P = .01), blood flow (P = .001), and blood volume (P = .002) at contrast-enhanced US than did patients who did not have myositis. Blood flow velocity (P = .001) and blood flow (P = .002) were significantly higher in patients with myositis than in volunteers. An increase in signal intensity on T2-weighted MR images was found in all patients with myositis, while contrast material enhancement on fat-suppressed T1-weighted MR images was found in only four of seven patients with myositis. CONCLUSION: Initial results show that contrast-enhanced US is a feasible method for noninvasively demonstrating increased perfusion in the involved muscle groups in patients with myositis. PMID- 16371586 TI - Subxiphoid access to normal pericardium with micropuncture set: technical feasibility study in pigs. AB - This study was performed with approval from the ethics committee for animal research of the local government. The purpose of the study was to evaluate the technical feasibility of a technique for subxiphoid access to the normal pericardial space with a micropuncture set in 10 large white pigs. With fluoroscopic guidance, a fine needle was inserted through a subxiphoid approach into the anterior mediastinal space to puncture the pericardium, and a micropuncture set was placed in the pericardial space successfully in all animals without complications. Necropsy at 24 hours did not reveal hemomediastinum, hemopericardium, or laceration of the pericardium. Results of the experiments in animals indicated that the technique was feasible and safe. PMID- 16371587 TI - The molecular motor KIFC1 associates with a complex containing nucleoporin NUP62 that is regulated during development and by the small GTPase RAN. AB - KIFC1 is a C-terminal kinesin motor associated with the nuclear membrane and acrosome in round and elongating spermatids. This location in developing spermatids is consistent with possible roles in acrosome elongation and manchette motility or both. Here we describe the association of the KIFC1 motor with a complex containing the nucleoporin NUP62. Formation of this complex is developmentally regulated, being absent before puberty and appearing only after nuclear elongation has begun. In addition, the integrity of this complex is dependent on GTP hydrolysis and the GTP state of the small GTPase RAN. Concomitant with the association of this motor with the NUP62-containing complex is an apparent reorganization of the nuclear pore with loss of NUP62 from larger complexes containing other nucleoporins. The association of KIFC1 with a component of the nuclear membrane is more consistent with a role for this motor in acrosome/manchette transport along the nuclear membrane than for a role for this motor in transport of vesicles along the outer face of the manchette. PMID- 16371588 TI - Meiosis in autologous ectopic transplants of immature testicular tissue grafted to Callithrix jacchus. AB - Grafting of immature testicular tissue provides a tool to examine testicular development and may offer a perspective for preservation of fertility in prepubertal patients. Successful xenografting in mice, resulting in mature spermatids, has been performed in several species but has failed with testicular tissues from the common marmoset, Callithrix jacchus. Previous data indicate that the hormonal milieu provided by the mouse host might cause this failure. We conducted autologous ectopic transplantation of testicular fragments under the back skin in newborn marmoset monkeys. Seventeen months after transplantation, we found viable transplants in 2 out of the 4 grafted animals. In the transplants, tubules developed up to a state intermediate between the pregraft situation and adult controls. Dividing spermatogonia and primary spermatocytes were present. Boule-like positivity and CDC25A negativity indicated that spermatogenesis was arrested at early meiosis. Immunohistochemistry revealed normal maturation of Sertoli cells, Leydig cells, and peritubular cells. Serum testosterone values were not restored to the normal range and bioactive chorionic gonadotropin levels increased to castrate levels. Meiotic arrest could have occurred in the grafts because of lack of sufficient testosterone or because of hyperthermia caused by the ectopic position of the grafts. We conclude that autologous transplants of immature testicular tissues in the marmoset can mature up to meiosis but that normal serum testosterone levels are not restored. Further studies have to be performed to overcome the meiotic arrest to explore the model further and to develop therapeutic options. PMID- 16371589 TI - Differential impact of intracellular carboxyl terminal domains on lipid raft localization of the murine gonadotropin-releasing hormone receptor. AB - The mammalian type I GNRH receptor (GNRHR) is unique among G protein-coupled receptors (GPCRs) because of the absence of an intracellular C-terminus. Previously, we have found that the murine GNRHR is constitutively localized to low-density membrane microdomains termed lipid rafts. As such, association of the GNRHR with lipid rafts may reflect both a loss (C-terminus) and a gain (raft association address) of structural characteristics. To address this, we fused either the full-length C-terminus from the nonraft-associated LH receptor (LHCGR; GNRHR-LF) or a truncated (t631) LHCGR C-terminus to the GNRHR. These chimeric receptors are trafficked to the plasma membrane, bind ligand, and display increased agonist-induced receptor internalization, but they do not partition into lipid rafts. Thus, a heterologous C-terminus from a nonraft-associated GPCR redirects localization of the GNRHR to nonraft domains. In contrast to the murine GNRHR, the catfish GNRHR (cfGNRHR) possesses an intracellular C-terminus. We found that the cfGNRHR was localized to lipid rafts and that the cfGNRHR C terminus did not alter raft localization of the mammalian receptor. Consistent with placement in different lipid microenvironments within the plasma membrane, fluorescence recovery after photobleaching revealed different lateral diffusion phenotypes of the raft-associated GNRHR and cfGNRHR versus the nonraft-associated GNRHR-LF fusion protein. We conclude that whereas an intracellular C-terminus is capable of redirecting the GNRHR to nonraft compartments, this is not a generalized feature of GPCR C-terminal tails. Thus, constitutive raft localization of the GNRHR is not simply a result of the loss of an intracellular C-terminus. PMID- 16371590 TI - Sperm from mice genetically deficient for the PCSK4 proteinase exhibit accelerated capacitation, precocious acrosome reaction, reduced binding to egg zona pellucida, and impaired fertilizing ability. AB - The gene for proprotein convertase subtilisin/kexin-like 4 (PCSK4, previously known as PC4) is primarily transcribed in testicular spermatogenic cells. Its inactivation in mouse causes severe male subfertility. To better understand the reproductive function of PCSK4, we examined its subcellular localization in the testicular epithelium via immunohistochemistry, and on intact sperm via indirect immunofluorescence and immunoelectron microscopy. PCSK4 was detected in the acrosomal granules of round spermatids, in the acrosomal ridges of elongated spermatids, and on the sperm plasma membrane overlying the acrosome. We also investigated PCSK4 relevance for sperm acquisition of fertilizing ability by comparing wild-type and PCSK4-null sperm for their abilities in capacitation, acrosome reaction, and egg binding in vitro. PCSK4-null sperm underwent capacitation at a faster rate; they were induced to acrosome react by lower concentrations of zona pellucida; and their egg-binding ability was only half that of wild-type sperm. These sperm physiologic anomalies likely contribute to the severe subfertility of PCSK4-deficient male mice. PMID- 16371591 TI - Regulation of the MAFF transcription factor by proinflammatory cytokines in myometrial cells. AB - The MAF (proto-)oncogene family of basic-leucine zipper transcription factors plays crucial roles in the control of mammalian gene expression and development. Here we analyzed the regulation of the human MAFF gene, coding for a small MAF transcription factor, in uterine smooth muscle cells. We found that MAFF transcript levels are induced by proinflammatory cytokines in PHM1-31 myometrial cells. We observed an important induction by interleukin 1 beta (IL1B) and a weaker upregulation by tumor necrosis factor (TNF), whereas interleukin 6 (IL6) treatment had no effect. Time course experiments revealed a rapid induction of MAFF transcripts within 30 min following IL1B treatment. The presence of actinomycin D inhibited the upregulation, suggesting that regulation of MAFF mRNA levels occurs at the transcriptional level. We generated a MAFF-specific antiserum and determined that MAFF protein was also induced by TNF and IL1B in PHM1-31 cells. In contrast, it was particularly interesting that the transcript and protein levels of the highly homologous MAFG and MAFK genes are not modulated by these cytokines. Our results suggest a possible specific role for MAFF in proinflammatory cytokine-mediated control of myometrial gene expression and provide the first link between a small MAF transcription factor and the inflammatory response. PMID- 16371592 TI - Nerve growth factor translates stress response and subsequent murine abortion via adhesion molecule-dependent pathways. AB - Spontaneous abortion is a frequent threat affecting 10%-25% of human pregnancies. Psychosocial stress has been suggested to be attributable for pregnancy losses by challenging the equilibrium of systems mandatory for pregnancy maintenance, including the nervous, endocrine, and immune system. Strong evidence indicates that stress-triggered abortion is mediated by adhesion molecules, i.e., intercellular adhesion molecule 1 (ICAM1) and leukocyte function associated molecule 1, now being referred to as integrin alpha L (ITGAL), which facilitate recruitment of inflammatory cells to the feto-maternal interface. The neurotrophin beta-nerve growth factor (NGFB), which has been shown to be upregulated in response to stress in multiple experimental settings including in the uterine lining (decidua) during pregnancy, increases ICAM1 expression on endothelial cells. Here, we investigated whether and how NGFB neutralization has a preventive effect on stress-triggered abortion in the murine CBA/J x DBA/2J model. We provide experimental evidence that stress exposure upregulates the frequency of abortion and the expression of uterine NGFB. Further, adhesion molecules ICAM1 and selectin platelet (SELP, formerly P-Selectin) and their ligands ITGAL and SELP ligand (SELPL, formerly P selectin glycoprotein ligand 1) respectively increase in murine deciduas in response to stress. Subsequently, decidual cytokines are biased toward a proinflammatory and abortogenic cytokine profile. Additionally, a decrease of pregnancy protective CD8alpha(+) decidual cells is present. Strikingly, all such uterine stress responses are abrogated by NGFB neutralization. Hence, NGFB acts as a proximal mediator in the hierarchical network of immune rejection by mediating an abortogenic environment comprised of classical signs of neurogenic inflammation. PMID- 16371593 TI - Effects of enucleation and caffeine on maturation-promoting factor (MPF) and mitogen-activated protein kinase (MAPK) activities in ovine oocytes used as recipient cytoplasts for nuclear transfer. AB - In general, oocytes arrested at metaphase of the second meiotic division (MII) are used as recipient cytoplasts for nuclear transfer (NT) procedures. MII oocytes contain high levels of maturation-promoting factor (MPF) and mitogen activated protein kinase (MAPK), which cause nuclear envelope breakdown (NEBD) and premature chromosome condensation (PCC) in the transferred nucleus and have been implicated in nuclear reprogramming. However, the occurrence of NEBD and the extent of PCC are variable between individual oocytes and species and are dependent on donor cell type and cell cycle stage. Enucleation, which removes oocyte cytoplasm, may reduce MPF and MAPK activities and reduce reprogramming; conversely, increasing kinase activities may increase reprogramming. We compared the effects of enucleation of ovine oocytes at anaphase/telophase of the first meiotic division (AI-TI) and at MII. MPF and MAPK activities were maximal at MII; blind enucleation at AI-TI was more efficient than at MII and removed a smaller volume of cytoplasm. Neither protocol significantly affected the activity of either kinase and the fate of the donor nucleus; however, enucleation per se significantly reduced the occurrence of NEBD in NT embryos. Treatment with 10 mM caffeine significantly increased the activities of both kinases and the occurrence of NEBD but did not affect the frequency of development to the blastocyst stage; however, a significant increase in total cell numbers was observed. The results show that caffeine can increase MPF and MAPK activities in ovine oocytes and that this may contribute to an increased reprogramming in NT embryos. PMID- 16371594 TI - Function of cGMP-dependent protein kinases as revealed by gene deletion. AB - Over the past few years, a wealth of biochemical and functional data have been gathered on mammalian cGMP-dependent protein kinases (cGKs). In mammals, three different kinases are encoded by two genes. Mutant and chimeric cGK proteins generated by molecular biology techniques yielded important biochemical knowledge, such as the function of the NH(2)-terminal domains of cGKI and cGKII, the identity of the cGMP-binding sites of cGKI, and the substrate specificity of the enzymes. Genetic approaches have proven especially useful for the analysis of the biological functions of cGKs. Recently, some of the in vivo targets and mechanisms leading to changes in neuronal adaptation, smooth muscle relaxation and growth, intestinal water secretion, bone growth, renin secretion, and other important functions have been identified. These data show that cGKs are signaling molecules involved in many biological functions. PMID- 16371596 TI - Dynamic sensorimotor interactions in locomotion. AB - Locomotion results from intricate dynamic interactions between a central program and feedback mechanisms. The central program relies fundamentally on a genetically determined spinal circuitry (central pattern generator) capable of generating the basic locomotor pattern and on various descending pathways that can trigger, stop, and steer locomotion. The feedback originates from muscles and skin afferents as well as from special senses (vision, audition, vestibular) and dynamically adapts the locomotor pattern to the requirements of the environment. The dynamic interactions are ensured by modulating transmission in locomotor pathways in a state- and phase-dependent manner. For instance, proprioceptive inputs from extensors can, during stance, adjust the timing and amplitude of muscle activities of the limbs to the speed of locomotion but be silenced during the opposite phase of the cycle. Similarly, skin afferents participate predominantly in the correction of limb and foot placement during stance on uneven terrain, but skin stimuli can evoke different types of responses depending on when they occur within the step cycle. Similarly, stimulation of descending pathways may affect the locomotor pattern in only certain phases of the step cycle. Section ii reviews dynamic sensorimotor interactions mainly through spinal pathways. Section iii describes how similar sensory inputs from the spinal or supraspinal levels can modify locomotion through descending pathways. The sensorimotor interactions occur obviously at several levels of the nervous system. Section iv summarizes presynaptic, interneuronal, and motoneuronal mechanisms that are common at these various levels. Together these mechanisms contribute to the continuous dynamic adjustment of sensorimotor interactions, ensuring that the central program and feedback mechanisms are congruous during locomotion. PMID- 16371597 TI - Sodium/calcium exchanger: influence of metabolic regulation on ion carrier interactions. AB - The Na(+)/Ca(2+) exchanger's family of membrane transporters is widely distributed in cells and tissues of the animal kingdom and constitutes one of the most important mechanisms for extruding Ca(2+) from the cell. Two basic properties characterize them. 1) Their activity is not predicted by thermodynamic parameters of classical electrogenic countertransporters (dependence on ionic gradients and membrane potential), but is markedly regulated by transported (Na(+) and Ca(2+)) and nontransported ionic species (protons and other monovalent cations). These modulations take place at specific sites in the exchanger protein located at extra-, intra-, and transmembrane protein domains. 2) Exchange activity is also regulated by the metabolic state of the cell. The mammalian and invertebrate preparations share MgATP in that role; the squid has an additional compound, phosphoarginine. This review emphasizes the interrelationships between ionic and metabolic modulations of Na(+)/Ca(2+) exchange, focusing mainly in two preparations where most of the studies have been carried out: the mammalian heart and the squid giant axon. A surprising fact that emerges when comparing the MgATP related pathways in these two systems is that although they are different (phosphatidylinositol bisphosphate in the cardiac and a soluble cytosolic regulatory protein in the squid), their final target effects are essentially similar: Na(+)-Ca(2+)-H(+) interactions with the exchanger. A model integrating both ionic and metabolic interactions in the regulation of the exchanger is discussed in detail as well as its relevance in cellular Ca(i)(2+) homeostasis. PMID- 16371598 TI - Skeletal muscle lipid metabolism in exercise and insulin resistance. AB - Lipids as fuel for energy provision originate from different sources: albumin bound long-chain fatty acids (LCFA) in the blood plasma, circulating very-low density lipoproteins-triacylglycerols (VLDL-TG), fatty acids from triacylglycerol located in the muscle cell (IMTG), and possibly fatty acids liberated from adipose tissue adhering to the muscle cells. The regulation of utilization of the different lipid sources in skeletal muscle during exercise is reviewed, and the influence of diet, training, and gender is discussed. Major points deliberated are the methods utilized to measure uptake and oxidation of LCFA during exercise in humans. The role of the various lipid-binding proteins in transmembrane and cytosolic transport of lipids is considered as well as regulation of lipid entry into the mitochondria, focusing on the putative role of AMP-activated protein kinase (AMPK), acetyl CoA carboxylase (ACC), and carnitine during exercise. The possible contribution to fuel provision during exercise of circulating VLDL-TG as well as the role of IMTG is discussed from a methodological point of view. The contribution of IMTG for energy provision may not be large, covering approximately 10% of total energy provision during fasting exercise in male subjects, whereas in females, IMTG may cover a larger proportion of energy delivery. Molecular mechanisms involved in breakdown of IMTG during exercise are also considered focusing on hormone-sensitive lipase (HSL). Finally, the role of lipids in development of insulin resistance in skeletal muscle, including possible molecular mechanisms involved, is discussed. PMID- 16371599 TI - Respiratory tract mucin genes and mucin glycoproteins in health and disease. AB - This review focuses on the role and regulation of mucin glycoproteins (mucins) in airway health and disease. Mucins are highly glycosylated macromolecules (> or =50% carbohydrate, wt/wt). MUC protein backbones are characterized by numerous tandem repeats that contain proline and are high in serine and/or threonine residues, the sites of O-glycosylation. Secretory and membrane-tethered mucins contribute to mucociliary defense, an innate immune defense system that protects the airways against pathogens and environmental toxins. Inflammatory/immune response mediators and the overproduction of mucus characterize chronic airway diseases: asthma, chronic obstructive pulmonary diseases (COPD), or cystic fibrosis (CF). Specific inflammatory/immune response mediators can activate mucin gene regulation and airway remodeling, including goblet cell hyperplasia (GCH). These processes sustain airway mucin overproduction and contribute to airway obstruction by mucus and therefore to the high morbidity and mortality associated with these diseases. Importantly, mucin overproduction and GCH, although linked, are not synonymous and may follow from different signaling and gene regulatory pathways. In section i, structure, expression, and localization of the 18 human MUC genes and MUC gene products having tandem repeat domains and the specificity and application of MUC-specific antibodies that identify mucin gene products in airway tissues, cells, and secretions are overviewed. Mucin overproduction in chronic airway diseases and secretory cell metaplasia in animal model systems are reviewed in section ii and addressed in disease-specific subsections on asthma, COPD, and CF. Information on regulation of mucin genes by inflammatory/immune response mediators is summarized in section iii. In section iv, deficiencies in understanding the functional roles of mucins at the molecular level are identified as areas for further investigations that will impact on airway health and disease. The underlying premise is that understanding the pathways and processes that lead to mucus overproduction in specific airway diseases will allow circumvention or amelioration of these processes. PMID- 16371600 TI - Signaling mechanisms regulating endothelial permeability. AB - The microvascular endothelial cell monolayer localized at the critical interface between the blood and vessel wall has the vital functions of regulating tissue fluid balance and supplying the essential nutrients needed for the survival of the organism. The endothelial cell is an exquisite "sensor" that responds to diverse signals generated in the blood, subendothelium, and interacting cells. The endothelial cell is able to dynamically regulate its paracellular and transcellular pathways for transport of plasma proteins, solutes, and liquid. The semipermeable characteristic of the endothelium (which distinguishes it from the epithelium) is crucial for establishing the transendothelial protein gradient (the colloid osmotic gradient) required for tissue fluid homeostasis. Interendothelial junctions comprise a complex array of proteins in series with the extracellular matrix constituents and serve to limit the transport of albumin and other plasma proteins by the paracellular pathway. This pathway is highly regulated by the activation of specific extrinsic and intrinsic signaling pathways. Recent evidence has also highlighted the importance of the heretofore enigmatic transcellular pathway in mediating albumin transport via transcytosis. Caveolae, the vesicular carriers filled with receptor-bound and unbound free solutes, have been shown to shuttle between the vascular and extravascular spaces depositing their contents outside the cell. This review summarizes and analyzes the recent data from genetic, physiological, cellular, and morphological studies that have addressed the signaling mechanisms involved in the regulation of both the paracellular and transcellular transport pathways. PMID- 16371601 TI - Microdomains of intracellular Ca2+: molecular determinants and functional consequences. AB - Calcium ions are ubiquitous and versatile signaling molecules, capable of decoding a variety of extracellular stimuli (hormones, neurotransmitters, growth factors, etc.) into markedly different intracellular actions, ranging from contraction to secretion, from proliferation to cell death. The key to this pleiotropic role is the complex spatiotemporal organization of the [Ca(2+)] rise evoked by extracellular agonists, which allows selected effectors to be recruited and specific actions to be initiated. In this review, we discuss the structural and functional bases that generate the subcellular heterogeneity in cellular Ca(2+) levels at rest and under stimulation. This complex choreography requires the concerted action of many different players; the central role is, of course, that of the calcium ion, with the main supporting characters being all the entities responsible for moving Ca(2+) between different compartments, while the cellular architecture provides a determining framework within which all the players have their exits and their entrances. In particular, we concentrate on the molecular mechanisms that lead to the generation of cytoplasmic Ca(2+) microdomains, focusing on their different subcellular location, mechanism of generation, and functional role. PMID- 16371604 TI - Intravenous iron and recombinant erythropoietin for the treatment of postoperative anemia. AB - PURPOSE: To determine if early recovery from severe post-operative anemia is accelerated by iv iron therapy alone or in combination with recombinant erythropoietin (EPO). METHODS: In this double-blinded, placebo-controlled randomized study, consenting adult patients without preoperative anemia whose hemoglobin concentration (Hb) was 70 to 90 g x L(-1) on the first day after cardiac or orthopedic surgery (POD 1) were assigned to one of three groups: control, iv iron alone (200 mg of iron sucrose on POD 1, 2, and 3) or in combination with EPO (600 U x kg(-1) on POD 1 and 3). The primary outcome was increase in Hb (adjusted for red blood cell transfusions) from POD 1 to 7. Analysis was by intention-to-treat in patients for whom the primary outcome was available. Group effect was analyzed by the ANOVA test, and between-group differences were specified with a Duncan multiple-range test. RESULTS: The primary outcome was available in 31 of 38 randomized patients. The average POD 1 Hb was 84 +/- 4 g x L(-1). There were no between-group differences in outcomes except for higher reticulocyte counts on POD-7 in the combination group. The average adjusted one-week increases in Hb were 7 +/- 8 g x L(-1) in the control group (n = 10), 9 +/- 9 g x L(-1) in the iv iron group (n = 11), and 10 +/- 14 g x L(-1) in the combination group (n = 10). The average adjusted six-week increases in Hb were 37 +/- 14 g x L(-1) in the control group, 40 +/- 7 g x L(-1) in the iv iron group, and 45 +/- 12 g x L(-1) in the combination group. CONCLUSION: Early postoperative treatment with iv iron alone or in combination with EPO does not appear to accelerate early recovery from postoperative anemia. PMID- 16371595 TI - Calcium at fertilization and in early development. AB - Fertilization calcium waves are introduced, and the evidence from which we can infer general mechanisms of these waves is presented. The two main classes of hypotheses put forward to explain the generation of the fertilization calcium wave are set out, and it is concluded that initiation of the fertilization calcium wave can be most generally explained in invertebrates by a mechanism in which an activating substance enters the egg from the sperm on sperm-egg fusion, activating the egg by stimulating phospholipase C activation through a src family kinase pathway and in mammals by the diffusion of a sperm-specific phospholipase C from sperm to egg on sperm-egg fusion. The fertilization calcium wave is then set into the context of cell cycle control, and the mechanism of repetitive calcium spiking in mammalian eggs is investigated. Evidence that calcium signals control cell division in early embryos is reviewed, and it is concluded that calcium signals are essential at all three stages of cell division in early embryos. Evidence that phosphoinositide signaling pathways control the resumption of meiosis during oocyte maturation is considered. It is concluded on balance that the evidence points to a need for phosphoinositide/calcium signaling during resumption of meiosis. Changes to the calcium signaling machinery occur during meiosis to enable the production of a calcium wave in the mature oocyte when it is fertilized; evidence that the shape and structure of the endoplasmic reticulum alters dynamically during maturation and after fertilization is reviewed, and the link between ER dynamics and the cytoskeleton is discussed. There is evidence that calcium signaling plays a key part in the development of patterning in early embryos. Morphogenesis in ascidian, frog, and zebrafish embryos is briefly described to provide the developmental context in which calcium signals act. Intracellular calcium waves that may play a role in axis formation in ascidian are discussed. Evidence that the Wingless/calcium signaling pathway is a strong ventralizing signal in Xenopus, mediated by phosphoinositide signaling, is adumbrated. The central role that calcium channels play in morphogenetic movements during gastrulation and in ectodermal and mesodermal gene expression during late gastrulation is demonstrated. Experiments in zebrafish provide a strong indication that calcium signals are essential for pattern formation and organogenesis. PMID- 16371605 TI - Bispectral index monitoring does not improve early recovery of geriatric outpatients undergoing brief surgical procedures. AB - PURPOSE: To assess if titration of sevoflurane using the bispectral index (BIS) monitor improves the early and intermediate recovery in geriatric outpatients undergoing brief urologic procedures under general anesthesia without muscle relaxants. METHODS: After a standardized induction with propofol and fentanyl, a laryngeal mask airway was inserted and sevoflurane was administered in combination with 60% nitrous oxide in oxygen for maintenance of anesthesia in spontaneously breathing outpatients. In the Control group (n = 25), sevoflurane and fentanyl were titrated according to standard clinical practice. In the BIS directed group (n = 25), sevoflurane was titrated to maintain a BIS value between 50 and 60, and supplemental fentanyl, 25 mug iv boluses were administered to treat tachypnea. The intraoperative anesthetic and analgesic requirements, as well as the times to eye opening, removal of the laryngeal mask airway device, response to simple commands, orientation to person and place, and postanesthesia care unit discharge eligibility (fast-track score of 14) were assessed at specific time intervals. RESULTS: The minimum alveolar concentration-hour of sevoflurane (0.25 +/- 0.15 and 0.31 +/- 0.2) and end-tidal concentrations of sevoflurane at the end of surgery (0.3 +/- 0.3 and 0.4 +/- 0.20%) did not differ significantly between the Control and BIS-directed groups, respectively. Although the percentage of patients requiring supplemental boluses of fentanyl was reduced in the BIS-directed group (16 vs 48%, P <0.05), the intraoperative BIS values and recovery times were similar in the two groups. CONCLUSION: In this non-paralyzed elderly outpatient surgery population, the use of BIS monitoring for titrating the maintenance anesthetic (sevoflurane) failed to improve the early recovery process. PMID- 16371606 TI - Audio spectrum and sound pressure levels vary between pulse oximeters. AB - PURPOSE: The variable-pitch pulse oximeter is an important intraoperative patient monitor. Our ability to hear its auditory signal depends on its acoustical properties and our hearing. This study quantitatively describes the audio spectrum and sound pressure levels of the monitoring tones produced by five variable-pitch pulse oximeters. METHODS: We compared the Datex-Ohmeda Capnomac Ultima, Hewlett-Packard M1166A, Datex-Engstrom AS/3, Ohmeda Biox 3700, and Datex Ohmeda 3800 oximeters. Three machines of each of the five models were assessed for sound pressure levels (using a precision sound level meter) and audio spectrum (using a hanning windowed fast Fourier trans-form of three beats at saturations of 99%, 90%, and 85%). RESULTS: The widest range of sound pressure levels was produced by the Hewlett-Packard M1166A (46.5 +/- 1.74 dB to 76.9 +/- 2.77 dB). The loudest model was the Datex-Engstrom AS/3 (89.2 +/- 5.36 dB). Three oximeters, when set to the lower ranges of their volume settings, were indistinguishable from background operating room noise. Each model produced sounds with different audio spectra. Although each model produced a fundamental tone with multiple harmonic overtones, the number of harmonics varied with each model; from three harmonic tones on the Hewlett-Packard M1166A, to 12 on the Ohmeda Biox 3700. There were variations between models, and individual machines of the same model with respect to the fundamental tone associated with a given saturation. CONCLUSION: There is considerable variance in the sound pressure and audio spectrum of commercially-available pulse oximeters. Further studies are warranted in order to establish standards. PMID- 16371607 TI - A multi source feedback program for anesthesiologists. AB - PURPOSE: To assess the feasibility, validity, and reliability of a multi source feedback program for anesthesiologists. METHODS: Surveys with 11, 19, 29 and 29 items were developed for patients, coworkers, medical colleagues and self, respectively, using five-point scales with an 'unable to assess' category. The items addressed communication skills, professionalism, collegiality, continuing professional development and collaboration. Each anesthesiologist was assessed by eight medical colleagues, eight coworkers, and 30 patients. Feasibility was assessed by response rates for each instrument. Validity was assessed by rating profiles, the percentage of participants unable to assess the physician for each item, and exploratory factor analyses to determine which items grouped together into scales. Cronbach's alpha and generalizability coefficient analyses assessed reliability. RESULTS: One hundred and eighty-six physicians participated. The mean number and percentage return rate of respondents per physician was 17.7 (56.2%) for patients, 7.8 (95.1%) for coworkers, and 7.8 (94.6%) for medical colleagues. The mean ratings ranged from four to five for each item on each scale. There were relatively few items with high percentages of 'unable to assess'. The factor analyses revealed a two-factor solution for the patient, a two-factor solution for the coworker and a three-factor solution for the medical colleague survey, accounting for at least 70% of the variance. All instruments had a high internal consistency reliability (Cronbach's alpha > 0.95). The generalizability coefficients were 0.65 for patients, 0.56 for coworkers and 0.69 for peers. CONCLUSION: It is feasible to develop multi source feedback instruments for anesthesiologists that are valid and reliable. PMID- 16371608 TI - Intraoperative somatosensory evoked potentials to facilitate peripheral nerve release. AB - PURPOSE: The significance of intraoperative somatosensory evoked potentials (SSEP) monitoring is well known during spinal surgery. This technology could be beneficial during peripheral nerve surgery as well. In order to illustrate potential applications, two cases of successful peripheral nerve release demonstrated by on-line, intraoperative, SSEP are reported. Clinical and technical features: The first case presents a complex brachial plexus lesion involving two mixed sensory-motor nerves: median and ulnar. The second case involved an entrapment neuropathy of the lateral femoral cutaneous nerve, a pure sensory nerve (meralgia paresthetica). For each patient we elicited specific peripheral nerve SSEP (recorded using bipolar cephalic montage) by stimulating each nerve independently. In each case, during difficult nerve dissection and after having excluded other possible factors of intraoperative SSEP variations, an increase of the SSEP amplitude was observed, and later correlated with favourable patient clinical outcome. CONCLUSIONS: Two cases demonstrate that intraoperative SSEP monitoring may provide an effective tool to guide surgical dissection during peripheral nerve release. This technique has potentially beneficial clinical applications and warrants further investigation. PMID- 16371609 TI - NSAID-analgesia, pain control and morbidity in cardiothoracic surgery. AB - OBJECTIVE: While narcotics remain the backbone of perioperative analgesia, the adjunctive role of other analgesics, including non-steroidal anti-inflammatory drugs (NSAIDs), is being recognized increasingly. This meta-analysis sought to determine whether adjunctive NSAIDs improve postoperative analgesia and reduce cumulative narcotic requirements. METHODS: A comprehensive search was undertaken to identify all randomized trials, in cardiothoracic patients, of NSAIDs plus narcotics vs narcotics without NSAIDs. Medline, Cochrane Library, EMBASE, and abstract databases were searched up to September 2005. The primary outcome was visual analogue scale (VAS) pain score. Secondary outcomes included 24-hr cumulative morphine-equivalents, rescue medications required, mortality, myocardial infarction, atrial fibrillation, stroke, renal failure, hospital readmissions, and in-hospital costs. RESULTS: Twenty randomized trials involving 1,065 patients were included. A significant reduction in 24-hr VAS pain score was found in patients receiving NSAIDs [weighted mean difference (WMD) -0.91 points, 95% confidence interval (CI) -1.48 to -0.34 points]. In addition, patients required significantly less morphine-equivalents in the first 24 hr (WMD -7.67 mg, 95% CI -8.97 to -6.38 mg). No significant difference was found with respect to mortality [odds ratio (OR) 0.19, 95% CI 0.01 to 4.22], myocardial infarction (OR 0.71, 95% CI 0.09 to 5.71), renal dysfunction (OR 0.95, 95% CI 0.37 to 2.46), or gastrointestinal bleeding (OR 0.96, 95% CI 0.13 to 7.09). CONCLUSION: In patients less than 70 yr of age undergoing cardiothoracic surgery, the adjunctive use of NSAIDs with narcotic analgesia reduces 24-hr VAS pain score and narcotic requirements. PMID- 16371610 TI - Intranasal sufentanil is effective for postoperative analgesia in adults. AB - PURPOSES: The aim of this prospective, randomized, double-blind study was to compare two doses of intranasal sufentanil for postoperative analgesia, titrated according to individual requirements based upon a numeric rating scale (NRS) from 0 to 10 for pain. METHODS: Forty patients, American Society of Anesthesiologists physical status I-II, scheduled for herniorrhaphy or hemorrhoidectomy under general anesthesia, were included when postoperative NRS was > 3. Nurses used a nasal puff device delivering a constant volume. Patients were randomized into two groups: Group A patients received a dose of 0.025 microg x kg(-1) /puff, Group B patients a dose of 0.05 microg x kg(-1) /puff. Puffs were administered as often as needed to obtain NRS < or = 3, with an interval time of five minutes. Hemodynamic, respiratory measures and sedation were recorded every five minutes. RESULTS: The probability of persistence of pain in Group B was consistently lower than in Group A. After 20 min, 20% of the patients had a NRS score > 3 in Group B, as opposed to 60% in Group A. At 60 min, no patient had a NRS > 3 in Group B, whereas there was a probability of 20% to record a NRS > 3 for Group A. Hemodynamic, respiratory parameters and sedation remained stable with no intergroup differences. CONCLUSIONS: Nasal administration of 0.050 microg x kg( 1) /puff sufentanil allowed a NRS < 4 to be attained within one hour in all patients, with efficacy achieved after 20 min. These findings suggest that the intranasal route is an effective mode of sufentanil administration for immediate postoperative analgesia in adult patients. PMID- 16371611 TI - Somatosensory evoked potentials as an objective assessment of the sensory median nerve blockade after infraclavicular block. AB - PURPOSE: Median nerve somatosensory evoked responses (MnSSER) alterations were compared to clinical tests (cold and pinprick) variations, in 20 ASA I adult patients following infraclavicular block obtained with 40 mL ropivacaine 0.5% to assess first, the difference of time course of the respective electrophysiological and clinical signs, and second, the objectivity and the reproducibility of MnSSER changes. CLINICAL FEATURES: Four MnSSER derivations (Erb's point; cutaneous projection of peripheral end of brachial plexus; posterior neck at C6 level, frontal and controlateral parietal scalp) were monitored and recorded for retrospective analysis. Continuous data acquisition were started before ropivacaine injection (baseline) and maintained for 30 min thereafter. Every three minutes after ropivacaine injection, cold and pinprick tests were performed in the hand median nerve cutaneous supply zone and were assessed using a sensory visual score (varying from 0-10). Data were compared using analysis of variance. Although MnSSER values were stable during baseline period, after ropivacaine administration, severe progressive amplitude depressions of selected MnSSER were detected in every patient. While clinical cold and pinprick tests became positive (score > 8) only 15.8 +/- 1.2 min and 20.1 +/- 1.8 min respectively after ropivacaine administration, the mean time to observe the earliest MnSSER 20% amplitude decrease at Erb's point derivation was reduced to 5.6 +/- 1.1 min (P < 0.01). CONCLUSION: Selected MnSSER amplitude reduction indicates objectively the onset of median nerve anesthesia following infraclavicular brachial plexus block before the appearance of clinical signs. PMID- 16371612 TI - High levels of impulsivity may contraindicate midazolam premedication in children. AB - PURPOSE: To investigate the effects of midazolam on emotional reactivity during induction of anesthesia in a pediatric day surgery setting. A secondary purpose was to determine if these effects were influenced by child temperament factors. METHODS: Forty children (age four to six years) scheduled for myringotomy were randomly assigned, in a double blind fashion, to receive either oral midazolam 0.5 mg.kg-1 mixed with acetaminophen suspension or acetaminophen alone. The Emotionality, Activity, Sociability, and Impulsivity (EASI) scale was used as a measure of child temperament. The modified Yale Preoperative Anxiety Scale (m YPAS), an observer-rated measure of state anxiety, was employed to assess anxiety pre- and post-drug, and also at induction of anesthesia. RESULTS: Children who received midazolam reacted significantly less to induction of anesthesia than did children in the placebo control group, F (1, 38) = 7.46, P = 0.01. A significant positive association was observed between baseline levels of anxiety and observer rated anxiety at anesthetic induction, but only in the placebo group, r = 0.58, P < 0.01. A significant positive association was observed between levels of impulsivity at baseline and observer-rated anxiety at anesthetic induction, but only in the midazolam group, r = 0.42, P < 0.05. CONCLUSIONS: Midazolam dampened adverse reactivity during anesthetic induction, particularly among children with high baseline levels of anxiety. Baseline level of impulsivity was positively associated with adverse reactions to anesthesia induction in the drug group, but not in the placebo group, suggesting that high levels of trait impulsivity may contraindicate the use of midazolam as a preoperative medication. PMID- 16371613 TI - A single dose of fentanyl and midazolam prior to Cesarean section have no adverse neonatal effects. AB - PURPOSE: Analgesia and sedation, routinely used as adjunct medications for regional anesthesia, are rarely used in the pregnant patient because of concerns about adverse neonatal effects. In an effort to obtain more information about maternal analgesia and sedation we studied neonatal and maternal effects of iv fentanyl and midazolam prior to spinal anesthesia for elective Cesarean section. METHODS: In this double-blinded, randomized, placebo-controlled trial, 60 healthy women received either a combination of 1 microg x kg(-1) fentanyl and 0.02 mg x kg(-1) midazolam intravenously or an equal volume of iv saline at the time of their skin preparation for a bupivacaine spinal anesthetic. Sample size was based on a non-parametric power analysis (power > 0.80 and alpha = 0.05) for clinically important differences in Apgar scores. Fetal outcome measures included Apgar scores, continuous pulse oximetry for three hours, and neurobehavioural scores. Maternal outcomes included catecholamine levels, and recall of anesthesia and delivery. RESULTS: There were no between-group differences of neonatal outcome variables (Apgar score, neurobehavioural scores, continuous oxygen saturation). Mothers in both groups showed no difference in their ability to recall the birth of their babies. CONCLUSIONS: Maternal analgesia and sedation with fentanyl (1 microg x kg(-1)) and midazolam (0.02 mg x kg(-1)) immediately prior to spinal anesthesia is not associated with adverse neonatal effects. PMID- 16371614 TI - When a leak is unavoidable, preoxygenation is equally ineffective with vital capacity or tidal volume breathing. AB - PURPOSE: Ideally, preoxygenation is performed using a tight fitting mask either by breathing normally for three to five minutes or with four to eight vital capacity (VC) breaths in 0.5 to one minute, but in practice leaks are frequent and sometimes unavoidable. This study was designed to determine which breathing method provided the best oxygenation in the presence of leak. METHODS: Twenty volunteers were instructed to breathe from a circle circuit supplied with 6 L x min(-1) of fresh oxygen. Each subject was tested under four situations selected in random order: 1) normal breathing for three minutes without leak; 2) normal breathing for three minutes with a leak; 3) four VCs in 30 sec without a leak; and 4) four VCs in 30 sec with a leak. The leak was created by a piece of size 18 French nasogastric tube, 5 cm long, taped under the face mask. Inspired and expired O(2) and CO(2) were sampled at the nostrils. RESULTS: In the absence of a leak, the end-tidal oxygen fraction (F(EO(2)) was greater after three minutes of tidal breathing (89 +/- 3%; mean +/- SD) in comparison with the response to four VCs (76 +/- 7%; P < 0.001). Introduction of a leak decreased the F(EO(2)) significantly (P < 0.001). With a leak, the F(EO(2)) was similar with normal breathing (61 +/- 8%) and after four VCs (59 +/- 11%). CONCLUSION: Preoxygenation with tidal volume breathing for three minutes yields higher F(EO(2)) in comparison to four VCs. If a small leak (4 mm internal diameter) is introduced, the F(EO(2)) decreases significantly with both breathing methods to approximately 60%. PMID- 16371615 TI - Innovative airway management for peritonsillar abscess. AB - PURPOSE: To describe innovative airway management in an adult with a peritonsillar abscess (quinsy) located atypically in the inferior pole of the palatine tonsil. CLINICAL FEATURES: A 25-yr-old male was admitted for surgical drainage of a left-sided, inferior pole peritonsillar abscess. Previous attempts at intraoral needle and scalpel drainage with topical anesthesia in the emergency department had failed. He had an interdental distance of 1.5 cm and computed tomography imaging showed narrowing of the airway diameter to 8 mm and lateral displacement of the epiglottis. He gargled 10 mL 0.5% lidocaine in the sitting position. We advanced a 3.1-mm pediatric fibreoptic bronchoscope (FOB) through an orally inserted nasopharyngeal airway to identify the glottis and sprayed 0.5% lidocaine onto the airway mucosa. We replaced the airway with a reinforced 6.5-mm internal diameter tracheal tube (TT), advanced the FOB through it until its tip was just above the carina, and then advanced the TT into the trachea. After iv induction of general anesthesia, the surgeon performed a tonsillectomy and drained the neck abscess. Postoperative direct laryngoscopy revealed a markedly improved airway lumen and tracheal extubation over a Cook Airway Exchange Catheter was uneventful. The patient was stable in the recovery room, and was discharged on the third postoperative day. CONCLUSION: We present an innovative technique of fibreoptic intubation in an awake patient with an inferior pole peritonsillar abscess. PMID- 16371616 TI - Transesophageal echocardiography images of anomalous circumflex coronary artery. PMID- 16371617 TI - Intraoperative jugular bulb desaturation during acute aneurysmal rupture. AB - PURPOSE: To describe an episode of acute jugular venous desaturation during intraoperative rupture of a cerebral aneurysm. CLINICAL FEATURES: A 57-yr-old patient was scheduled for clipping of a large unruptured basilar tip aneurysm. Abrupt bulging of the brain was observed after bone flap removal, but before dura was opened. This was associated with concurrent development of systemic hypertension to 200/120 mmHg and jugular venous bulb (S(jv)O(2)) desaturation to 13%. Rupture of aneurysm was confirmed by frank blood in cerebrospinal fluid drainage from the lumbar subarachnoid catheter. CONCLUSIONS: Abrupt S(jv)O(2) desaturation prior to dural opening may suggest an acute increase in intracranial pressure, which in our case followed aneurysmal rupture; the systemic response to increased intracranial pressure (Cushing's response) may be ineffective in maintaining cerebral perfusion. PMID- 16371618 TI - Management of acute respiratory distress syndrome using pumpless extracorporeal lung assist. AB - PURPOSE: To describe the use of a pumpless extracorporeal lung assist device in the treatment of severe acute respiratory distress syndrome (ARDS). CLINICAL FEATURES: A 15-yr-old girl developed severe post-traumatic ARDS. After all conventional treatment strategies failed, we inserted a pumpless extracorporeal lung assist device. This device consists of an arterial cannula inserted into the femoral artery, and a membrane oxygenator with a venous cannula that returns the oxygenated blood back to the patient's femoral vein. Since the driving force is the patient's blood pressure, a roller pump with its negative side effects is not needed. The device allowed removal of excessive PaCO(2) and, by applying minimal ventilation, minimization of further ventilator-induced lung injury. The pumpless extracorporeal lung assist device remained in situ for ten days without any adverse side effect. During this time, the lung recovered such that mechanical ventilation could be reinstalled cautiously. The device was then removed and, after a prolonged period of intensive care, the patient recovered without any sequelae. CONCLUSION: In this case of a severely damaged lung, an arteriovenous pumpless extracorporeal lung assist was a helpful device to remove elevated CO(2) and reduce mechanical stress by applying minimal ventilation. This device is simple to operate and has the potential of being used routinely in the treatment of severe ARDS. PMID- 16371619 TI - Cuffed versus uncuffed pediatric endotracheal tubes. PMID- 16371620 TI - Complication of a non-invasive blood pressure cuff during prone positioning. PMID- 16371621 TI - Proper preparation of the Trachlight and endotracheal tube to facilitate intubation. PMID- 16371622 TI - Improving patient safety by design -- a new spinal/intrathecal injection safety system. PMID- 16371623 TI - Ropivacaine 0.5% administered via a femoral catheter: is the obturator nerve also blocked? PMID- 16371624 TI - Delayed respiratory depression after interscalene blockade for shoulder surgery in geriatric patients. PMID- 16371625 TI - Images in clinical medicine. Eye worm. PMID- 16371626 TI - Oseltamivir resistance--disabling our influenza defenses. PMID- 16371627 TI - The run on tamiflu--should physicians prescribe on demand? PMID- 16371628 TI - Safety of long-acting beta-agonists--an urgent need to clear the air. PMID- 16371629 TI - Tumor-infiltrating T cells--friend or foe of neoplastic cells? PMID- 16371630 TI - Intensive diabetes treatment and cardiovascular disease in patients with type 1 diabetes. AB - BACKGROUND: Intensive diabetes therapy aimed at achieving near normoglycemia reduces the risk of microvascular and neurologic complications of type 1 diabetes. We studied whether the use of intensive therapy as compared with conventional therapy during the Diabetes Control and Complications Trial (DCCT) affected the long-term incidence of cardiovascular disease. METHODS: The DCCT randomly assigned 1441 patients with type 1 diabetes to intensive or conventional therapy, treating them for a mean of 6.5 years between 1983 and 1993. Ninety three percent were subsequently followed until February 1, 2005, during the observational Epidemiology of Diabetes Interventions and Complications study. Cardiovascular disease (defined as nonfatal myocardial infarction, stroke, death from cardiovascular disease, confirmed angina, or the need for coronary-artery revascularization) was assessed with standardized measures and classified by an independent committee. RESULTS: During the mean 17 years of follow-up, 46 cardiovascular disease events occurred in 31 patients who had received intensive treatment in the DCCT, as compared with 98 events in 52 patients who had received conventional treatment. Intensive treatment reduced the risk of any cardiovascular disease event by 42 percent (95 percent confidence interval, 9 to 63 percent; P=0.02) and the risk of nonfatal myocardial infarction, stroke, or death from cardiovascular disease by 57 percent (95 percent confidence interval, 12 to 79 percent; P=0.02). The decrease in glycosylated hemoglobin values during the DCCT was significantly associated with most of the positive effects of intensive treatment on the risk of cardiovascular disease. Microalbuminuria and albuminuria were associated with a significant increase in the risk of cardiovascular disease, but differences between treatment groups remained significant (P< or =0.05) after adjusting for these factors. CONCLUSIONS: Intensive diabetes therapy has long-term beneficial effects on the risk of cardiovascular disease in patients with type 1 diabetes. PMID- 16371631 TI - Effector memory T cells, early metastasis, and survival in colorectal cancer. AB - BACKGROUND: The role of tumor-infiltrating immune cells in the early metastatic invasion of colorectal cancer is unknown. METHODS: We studied pathological signs of early metastatic invasion (venous emboli and lymphatic and perineural invasion) in 959 specimens of resected colorectal cancer. The local immune response within the tumor was studied by flow cytometry (39 tumors), low-density array real-time polymerase-chain-reaction assay (75 tumors), and tissue microarrays (415 tumors). RESULTS: Univariate analysis showed significant differences in disease-free and overall survival according to the presence or absence of histologic signs of early metastatic invasion (P<0.001). Multivariate Cox analysis showed that an early conventional pathological tumor-node-metastasis stage (P<0.001) and the absence of early metastatic invasion (P=0.04) were independently associated with increased survival. As compared with tumors with signs of early metastatic invasion, tumors without such signs had increased infiltrates of immune cells and increased levels of messenger RNA (mRNA) for products of type 1 helper effector T cells (CD8, T-BET [T-box transcription factor 21], interferon regulatory factor 1, interferon-gamma, granulysin, and granzyme B) but not increased levels of inflammatory mediators or immunosuppressive molecules. The two types of tumors had significant differences in the levels of expression of 65 combinations of T-cell markers, and hierarchical clustering showed that markers of T-cell migration, activation, and differentiation were increased in tumors without signs of early metastatic invasion. The latter type of tumors also had increased numbers of CD8+ T cells, ranging from early memory (CD45RO+CCR7-CD28+CD27+) to effector memory (CD45RO+CCR7-CD28-CD27-) T cells. The presence of high levels of infiltrating memory CD45RO+ cells, evaluated immunohistochemically, correlated with the absence of signs of early metastatic invasion, a less advanced pathological stage, and increased survival. CONCLUSIONS: Signs of an immune response within colorectal cancers are associated with the absence of pathological evidence of early metastatic invasion and with prolonged survival. PMID- 16371632 TI - Oseltamivir resistance during treatment of influenza A (H5N1) infection. AB - Influenza A (H5N1) virus with an amino acid substitution in neuraminidase conferring high-level resistance to oseltamivir was isolated from two of eight Vietnamese patients during oseltamivir treatment. Both patients died of influenza A (H5N1) virus infection, despite early initiation of treatment in one patient. Surviving patients had rapid declines in the viral load to undetectable levels during treatment. These observations suggest that resistance can emerge during the currently recommended regimen of oseltamivir therapy and may be associated with clinical deterioration and that the strategy for the treatment of influenza A (H5N1) virus infection should include additional antiviral agents. PMID- 16371633 TI - Disciplinary action by medical boards and prior behavior in medical school. AB - BACKGROUND: Evidence supporting professionalism as a critical measure of competence in medical education is limited. In this case-control study, we investigated the association of disciplinary action against practicing physicians with prior unprofessional behavior in medical school. We also examined the specific types of behavior that are most predictive of disciplinary action against practicing physicians with unprofessional behavior in medical school. METHODS: The study included 235 graduates of three medical schools who were disciplined by one of 40 state medical boards between 1990 and 2003 (case physicians). The 469 control physicians were matched with the case physicians according to medical school and graduation year. Predictor variables from medical school included the presence or absence of narratives describing unprofessional behavior, grades, standardized-test scores, and demographic characteristics. Narratives were assigned an overall rating for unprofessional behavior. Those that met the threshold for unprofessional behavior were further classified among eight types of behavior and assigned a severity rating (moderate to severe). RESULTS: Disciplinary action by a medical board was strongly associated with prior unprofessional behavior in medical school (odds ratio, 3.0; 95 percent confidence interval, 1.9 to 4.8), for a population attributable risk of disciplinary action of 26 percent. The types of unprofessional behavior most strongly linked with disciplinary action were severe irresponsibility (odds ratio, 8.5; 95 percent confidence interval, 1.8 to 40.1) and severely diminished capacity for self-improvement (odds ratio, 3.1; 95 percent confidence interval, 1.2 to 8.2). Disciplinary action by a medical board was also associated with low scores on the Medical College Admission Test and poor grades in the first two years of medical school (1 percent and 7 percent population attributable risk, respectively), but the association with these variables was less strong than that with unprofessional behavior. CONCLUSIONS: In this case-control study, disciplinary action among practicing physicians by medical boards was strongly associated with unprofessional behavior in medical school. Students with the strongest association were those who were described as irresponsible or as having diminished ability to improve their behavior. Professionalism should have a central role in medical academics and throughout one's medical career. PMID- 16371634 TI - Inhaled nitric oxide therapy in adults. PMID- 16371635 TI - Images in clinical medicine. Gastric dilatation secondary to diabetic autonomic neuropathy. PMID- 16371636 TI - Case records of the Massachusetts General Hospital. Case 39-2005. A 63-year-old woman with a positive serologic test for syphilis and persistent eosinophilia. PMID- 16371637 TI - Glycemic control and cardiovascular disease--should we reassess clinical goals? PMID- 16371638 TI - Professional behavior--a learner's permit for licensure. PMID- 16371639 TI - Immunosuppression, skin cancer, and ultraviolet A radiation. PMID- 16371640 TI - Management of acute coronary syndromes. PMID- 16371641 TI - Protective conditioning for acute graft-versus-host disease. PMID- 16371642 TI - Primary biliary cirrhosis. PMID- 16371643 TI - Academic-industrial relationships. PMID- 16371644 TI - Imatinib and hyperlipidemia. PMID- 16371645 TI - Sphingosine 1-phosphate is released from the cytosol of rat platelets in a carrier-mediated manner. AB - Sphingosine 1-phosphate (S1P) is accumulated in platelets and released on stimulation by thrombin or Ca(2+). Thrombin-stimulated S1P release was inhibited by staurosporin, whereas Ca(2+)-stimulated release was not. When the platelet plasma membrane was permeabilized with streptolysin O (SLO), S1P leaked out with cytosol markers, whereas granular markers remained in the platelets. The SLO induced S1P leakage required BSA, probably for solubilization of S1P in the medium. These results indicate that S1P is localized in the inner leaflet of the plasma membrane and that its release is a carrier-mediated process. We also used alpha-toxin (ATX), which makes smaller pores in the plasma membrane than SLO and depletes cytosolic ATP without BSA-dependent S1P leakage. The addition of ATP drove S1P release from ATX platelets. The ATP-driven S1P release from ATX platelets was greatly enhanced by thrombin. An ATP binding cassette transporter inhibitor, glyburide, prevents ATP- and thrombin-induced S1P release from platelets. Ca(2+) also stimulated S1P release from ATX platelets without ATP, whereas the Ca(2+)-induced release was not inhibited by glyburide. Our results indicate that two independent S1P release systems might exist in the platelet plasma membrane, an ATP-dependent system stimulated by thrombin and an ATP independent system stimulated by Ca(2+). PMID- 16371646 TI - Interaction between mild hypercholesterolemia, HDL-cholesterol levels, and angiotensin II in intimal hyperplasia in mice. AB - Two month old C57BL/6 mice were placed on three different diets: 1) normal diet (NC; 0.025% cholesterol), 2) hypercholesterolemic Western-type diet (HC-W; 0.2% cholesterol), and 3) hypercholesterolemic Paigen-type diet (HC-P; 1.25% cholesterol plus 0.5% cholic acid). At 6 months of age, the animals underwent ligation of the left carotid artery and were randomly assigned to vehicle (PBS, subcutaneous) or angiotensin II (Ang II; 1.4 mg/kg/day, subcutaneous) treatment for 4 weeks. Low density lipoprotein-cholesterol levels were similarly increased in both HC diets (NC, 4 +/- 3 mg/dl; HC-W, 123 +/- 17 mg/dl; HC-P, 160 +/- 14 mg/dl). However, the levels of high density lipoprotein-cholesterol (HDL-C) were reduced only in animals fed the HC-P diet (NC, 82 +/- 6 mg/dl; HC-W, 79 +/- 7 mg/dl; HC-P, 58 +/- 7 mg/dl). In Ang II-treated mice, carotid artery ligation induced intimal smooth muscle cell proliferation to a similar extent in NC- and HP-W-fed animals. However, a significantly larger intimal area developed in ligated vessels from Ang II-treated mice fed the HC-P diet (3.6-fold higher than in Ang II-treated NC mice). Together, these results show the accelerating effect of mild hypercholesterolemia, reduced HDL-C levels, and Ang II on intimal hyperplasia after carotid artery ligation in mice. PMID- 16371647 TI - Rapid, specific, and sensitive measurements of plasma sphingomyelin and phosphatidylcholine. AB - Sphingomyelin (SM) and phosphatidylcholine (PC) are two major phospholipids on plasma lipoproteins. Their concentration is classically measured by lipid extraction, thin-layer chromatography, and phosphate determination on separated SM or PC spots. Here, we describe two rapid, specific, and sensitive enzymatic measurements for both phospholipids. Plasma was incubated with bacterial sphingomyelinase (for SM measurement) or bacterial PC-specific phospholipase D (for PC measurement), alkaline phosphatase, choline oxidase, peroxidase, N-ethyl N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, and 4-aminoantipyrine for 45 min. A blue dye, with an optimal absorption at 595 nm, was generated. PC levels did not influence SM measurement and vice versa. The linear range for the SM measurement was 0.5-5 microg, and that for PC was 2.5-20 microg. The mean percentage recovery was 98.0 +/- 5.2% for SM and 96.6 +/- 3.8% for PC. The interassay coefficient of variation of the assay was 1.7 +/- 0.05% for SM and 3.1 +/- 0.13% for PC. These two new methods are amenable to automation and can be adapted for large-scale, high-throughput assays. PMID- 16371648 TI - Selecting against hypotheses. By Caveman. PMID- 16371649 TI - Here come the septins: novel polymers that coordinate intracellular functions and organization. AB - Septins are conserved GTP-binding proteins that associate with cellular membranes and the actin and microtubule cytoskeletons. They polymerize to form filamentous structures that act as diffusion barriers between different membrane domains and as molecular scaffolds for membrane- and cytoskeleton-binding proteins. In yeast, septins are central to the spatio-temporal coordination of membrane polarity and cell division, but the roles of their mammalian counterparts have remained poorly understood. However, recent findings have shed light on the dynamics and regulation of mammalian septin assembly and our understanding of septin functions in cytoskeleton and membrane organization. The mammalian septins appear to form a novel network of hetero-polymers that are multi-functional, inter-changeable and respond dynamically to signals that coordinate events at the interface between cytoskeleton and membrane biology. Hence, studies of these molecules might provide new insights not only into how cells coordinate their functions, but also into the pathogenesis of cancer and other diseases in which septins are abnormally expressed. PMID- 16371651 TI - Single-molecule analysis of cadherin-mediated cell-cell adhesion. AB - Cadherins are ubiquitous cell surface molecules that are expressed in virtually all solid tissues and localize at sites of cell-cell contact. Cadherins form a large and diverse family of adhesion molecules, which play a crucial role in a multitude of cellular processes, including cell-cell adhesion, motility, and cell sorting in maturing organs and tissues, presumably because of their different binding capacity and specificity. Here, we develop a method that probes the biochemical and biophysical properties of the binding interactions between cadherins expressed on the surface of living cells, at the single-molecule level. Single-molecule force spectroscopy reveals that classical cadherins, N-cadherin and E-cadherin, form bonds that display adhesion specificity, and a pronounced difference in adhesion force and reactive compliance, but not in bond lifetime. Moreover, their potentials of interaction, derived from force-spectroscopy measurements, are qualitatively different when comparing the single-barrier energy potential for the dissociation of an N-cadherin-N-cadherin bond with the double-barrier energy potential for an E-cadherin-E-cadherin bond. Together these results suggest that N-cadherin and E-cadherin molecules form homophilic bonds between juxtaposed cells that have significantly different kinetic and micromechanical properties. PMID- 16371650 TI - FAK and Src kinases are required for netrin-induced tyrosine phosphorylation of UNC5. AB - During neuronal development, netrin and its receptors UNC5 and DCC (deleted in colorectal cancer) guide axonal growth cones in navigating to their targets. Netrin also plays important roles in the regulation of cell migration, tissue morphogenesis and tumor growth. Here, we show that netrin induces UNC5 tyrosine phosphorylation and that this effect of netrin is dependent on its co-receptor DCC. UNC5 tyrosine phosphorylation is known to be important for netrin to induce cell migration and axonal repulsion. Src tyrosine kinase activity is required for netrin to stimulate UNC5 tyrosine phosphorylation in neurons and transfected cells. The SH2 domain of Src kinase directly interacts with the cytosolic domain of UNC5 in a tyrosine-phosphorylation-dependent manner. Furthermore, the tyrosine kinase focal adhesion kinase (FAK) is also involved in netrin-induced UNC5 tyrosine phosphorylation. Both Src and FAK can phosphorylate UNC5. Our data suggest a model in which netrin stimulates UNC5 tyrosine phosphorylation and signaling in a manner dependent on the co-receptor DCC, through the recruitment of Src and FAK kinases. PMID- 16371652 TI - Production of reactive oxygen species in response to replication stress and inappropriate mitosis in fission yeast. AB - Previous studies have indicated that replication stress can trigger apoptosis like cell death, accompanied (where tested) by production of reactive oxygen species (ROS), in mammalian cells and budding yeast (Saccharomyces cerevisiae). In mammalian cells, inappropriate entry into mitosis also leads to cell death. Here, we report similar responses in fission yeast (Schizosaccharomyces pombe). We used ROS- and death-specific fluorescent stains to measure the effects of mutations in replication initiation and checkpoint genes in fission yeast on the frequencies of ROS production and cell death. We found that certain mutant alleles of each of the four tested replication initiation genes caused elevated ROS and cell death. Where tested, these effects were not enhanced by checkpoint gene mutations. Instead, when cells competent for replication but defective in both the replication and damage checkpoints were treated with hydroxyurea, which slows replication fork movement, the frequencies of ROS production and cell death were greatly increased. This was a consequence of elevated CDK activity, which permitted inappropriate entry into mitosis. Thus, studies in fission yeast are likely to prove helpful in understanding the pathways that lead from replication stress and inappropriate mitosis to cell death in mammalian cells. PMID- 16371653 TI - Neural induction promotes large-scale chromatin reorganisation of the Mash1 locus. AB - Determining how genes are epigenetically regulated to ensure their correct spatial and temporal expression during development is key to our understanding of cell lineage commitment. Here we examined epigenetic changes at an important proneural regulator gene Mash1 (Ascl1), as embryonic stem (ES) cells commit to the neural lineage. In ES cells where the Mash1 gene is transcriptionally repressed, the locus replicated late in S phase and was preferentially positioned at the nuclear periphery with other late-replicating genes (Neurod, Sprr2a). This peripheral location was coupled with low levels of histone H3K9 acetylation at the Mash1 promoter and enhanced H3K27 methylation but surprisingly location was not affected by removal of the Ezh2/Eed HMTase complex or several other chromatin silencing candidates (G9a, SuV39h-1, Dnmt-1, Dnmt-3a and Dnmt-3b). Upon neural induction however, Mash1 transcription was upregulated (>100-fold), switched its time of replication from late to early in S phase and relocated towards the interior of the nucleus. This spatial repositioning was selective for neural commitment because Mash1 was peripheral in ES-derived mesoderm and other non neural cell types. A bidirectional analysis of replication timing across a 2 Mb region flanking the Mash1 locus showed that chromatin changes were focused at Mash1. These results suggest that Mash1 is regulated by changes in chromatin structure and location and implicate the nuclear periphery as an important environment for maintaining the undifferentiated state of ES cells. PMID- 16371654 TI - Ubiquitin C-terminal hydrolase L1 regulates the morphology of neural progenitor cells and modulates their differentiation. AB - Ubiquitin C-terminal hydrolase L1 (UCH-L1) is a component of the ubiquitin system, which has a fundamental role in regulating various biological activities. However, the functional role of the ubiquitin system in neurogenesis is not known. Here we show that UCH-L1 regulates the morphology of neural progenitor cells (NPCs) and mediates neurogenesis. UCH-L1 was expressed in cultured NPCs as well as in embryonic brain. Its expression pattern in the ventricular zone (VZ) changed between embryonic day (E) 14 and E16, which corresponds to the transition from neurogenesis to gliogenesis. At E14, UCH-L1 was highly expressed in the ventricular zone, where neurogenesis actively occurs; whereas its expression was prominent in the cortical plate at E16. UCH-L1 was very weakly detected in the VZ at E16, which corresponds to the start of gliogenesis. In cultured proliferating NPCs, UCH-L1 was co-expressed with nestin, a marker of undifferentiated cells. In differentiating cells, UCH-L1 was highly co-expressed with the early neuronal marker TuJ1. Furthermore, when UCH-L1 was induced in nestin-positive progenitor cells, the number and length of cellular processes of the progenitors decreased, suggesting that the progenitor cells were differentiating. In addition, NPCs derived from gad (UCH-L1-deficient) mice had longer processes compared with controls. The ability of UCH-L1 to regulate the morphology of nestin-positive progenitors was dependent on its binding affinity for ubiquitin but not on hydrolase activity; this result was also confirmed using gad-mouse-derived NPCs. These results suggest that UCH-L1 spatially mediates and enhances neurogenesis in the embryonic brain by regulating progenitor cell morphology. PMID- 16371655 TI - Functional analysis of SIRPalpha in the growth cone. AB - The 'signal regulatory protein' SIRPalpha is an Ig superfamily, transmembrane glycoprotein with a pair of cytoplasmic domains that can bind the phosphatase SHP 2 when phosphorylated on tyrosine. SIRPalpha is prominent in growth cones of rat cortical neurons and located, together with the tetraspanin CD81, in the growth cone periphery. SIRPalpha is dynamically associated with Triton-X-100-sensitive, but Brij-98-resistant, lipid microdomains, which also contain CD81. Challenge of growth cones with the integrin-binding extracellular-matrix (ECM) protein, laminin, or with the growth factors, IGF-1 or BDNF, increases SIRPalpha phosphorylation and SHP-2 binding rapidly and transiently, via Src family kinase activation; phosphorylated SIRPalpha dissociates from the lipid microdomains. A cytoplasmic tail fragment of SIRPalpha (cSIRPalpha), when expressed in primary cortical neurons, also is phosphorylated and binds SHP-2. Expression of wild-type cSIRPalpha, but not of a phosphorylation-deficient mutant, substantially decreases IGF-1-stimulated axonal growth on laminin. On poly-D-lysine and in control conditions, axonal growth is slower than on laminin, but there is no further reduction in growth rate induced by the expression of cSIRPalpha. Thus, the effect of cSIRPalpha on axon growth is dependent upon integrin activation by laminin. These results suggest that SIRPalpha functions in the modulation of axonal growth by ECM molecules, such as laminin. PMID- 16371656 TI - Myo10 in brain: developmental regulation, identification of a headless isoform and dynamics in neurons. AB - Although Myo10 (myosin-X) is an unconventional myosin associated with filopodia, little is known about its isoforms and roles in the nervous system. We report here that, in addition to full-length Myo10, brain expresses a shorter form of Myo10 that lacks a myosin head domain. This ;headless' Myo10 is thus unable to function as a molecular motor, but is otherwise identical to full-length Myo10 and, like it, contains three pleckstrin homology (PH) domains, a myosin-tail homology 4 (MyTH4) domain, and a band-4.1/ezrin/radixin/moesin (FERM) domain. Immunoblotting demonstrates that both full-length and headless Myo10 exhibit dramatic developmental regulation in mouse brain. Immunofluorescence with an antibody that detects both isoforms demonstrates that Myo10 is expressed in neurons, such as Purkinje cells, as well as non-neuronal cells, such as astrocytes and ependymal cells. CAD cells, a neuronal cell line, express both full-length and headless Myo10, and this endogenous Myo10 is present in cell bodies, neurites, growth cones and the tips of filopodia. To investigate the dynamics of the two forms of Myo10 in neurons, CAD cells were transfected with GFP constructs corresponding to full-length or headless Myo10. Only full-length Myo10 localizes to filopodial tips and undergoes intrafilopodial motility, demonstrating that the motor domain is necessary for these activities. Live cell imaging also reveals that full-length Myo10 localizes to the tips of neuronal filopodia as they explore and interact with their surroundings, suggesting that this myosin has a role in neuronal actin dynamics. PMID- 16371657 TI - Loose chromosomes sink cells. PMID- 16371658 TI - Harnessing hormonal signaling for cardioprotection. AB - Cardiovascular disease is the leading cause of death in women in the Western world and is predominant among the elderly. A large body of evidence suggests that hormonal signaling plays a critical role in the regulation of cardioprotective mechanisms, as premenopausal women are at significantly lower risk of heart disease compared with men, but the risk greatly increases with the onset of menopause. This association indicates that estrogen may protect the heart from cardiovascular disease. Whereas a number of analyses of the effects of hormone replacement therapy (HRT) on postmenopausal women supported the idea that estrogen is a cardioprotective factor, the findings of the more recent Women's Health Initiative (WHI) study suggested that HRT may actually increase the risk of cardiovascular events. These conflicting reports have left both patients and clinicians reluctant to continue using current HRT regimes. The WHI findings do not, however, negate the epidemiological link between menopause and increased cardiovascular risk. Hence, the identification of the specific actions of estrogen that promote cardioprotective pathways without enhancing deleterious vascular mechanisms may provide novel estrogen-based alternatives to current HRT strategies. In this Review, we outline the known actions of estrogen on the cardiovascular system, focusing on cardioprotective mechanisms that may be targeted for the development of new therapeutic approaches. PMID- 16371659 TI - Coaching as a treatment for ADHD. PMID- 16371660 TI - Serum cholinesterase levels elevated in patients suffering from ADHD. PMID- 16371661 TI - What is the prevalence of adult ADHD? Results of a population screen of 966 adults. AB - To provide a better estimate of the prevalence of ADHD in adulthood, the authors complete a telephone survey of 966 randomly selected adults. They compute two diagnoses from the survey data. Participants meeting Diagnostic and Statistical Manual of Mental Disorders (4th ed.) criteria for both childhood and adulthood are defined as narrow ADHD. Broad ADHD adds to that definition those meeting subthreshold criteria. Cronbach's alpha is .90 for the 18 DSM-IV symptoms in childhood and .88 when rated for current symptoms in adulthood. No one item unduly influences the reliability of the total score. The authors find similar results in separate analyses of hyperactive-impulsive and inattentive symptoms. They estimate prevalences of 2.9% for Narrow ADHD and 16.4% for Broad ADHD. Having ADHD is associated with lower levels of education and employment status. These findings suggest that adult ADHD is a common disorder associated with impaired functioning. PMID- 16371662 TI - Prevalence and correlates of ADHD symptoms in the national health interview survey. AB - OBJECTIVE: Study the prevalence and correlates of ADHD symptoms in the National Health Interview Survey (NHIS). METHODS: NHIS includes 10,367 children ages 4 to 17. Parents report lifetime diagnosis of ADHD and complete the Strengths and Difficulties Questionnaire (SDQ). Prevalences of clinically significant ADHD and comorbid symptoms by race and ethnicity, gender, and age are presented. RESULTS: Prevalence of clinically significant SDQ ADHD symptoms is 4.19% (males) and 1.77% (females). Male prevalence by race is 3.06% for Hispanics, 4.33% for Whites, and 5.65% for Blacks. Significant differences in prevalence occur across gender (p < .01) and among males across race (p < .01), age (p < .01), and income (p < .02). In the full sample, 6.80% of males and 2.50% of females have a parent-reported lifetime ADHD diagnosis but are negative for SDQ ADHD. Likewise, 1.59% of males and 0.81% of females are positive for SDQ ADHD but negative for parent report of ADHD diagnosis. SDQ ADHD positive children have substantially higher proportions of elevated scores on other SDQ subscales. CONCLUSION: ADHD symptoms vary by race and ethnicity, gender, and age and are associated with other emotional and behavioral difficulties. Both overdiagnosis and underdiagnosis of ADHD may be issues in the U.S. population of 4- to 17-year-olds. PMID- 16371663 TI - Assessment of ADHD: differences across psychology specialty areas. AB - Child psychologists are frequently involved in the assessment of ADHD symptoms among school-aged youth. There is limited information regarding the extent to which psychologists adhere to recommended assessment practices and whether differences exist in assessment strategies among psychologists from different specialty areas (clinical, counseling, and school) and/or who practice in different settings (university, school, or outpatient clinic). A 3 (specialty area) x 3 (employment setting) between-groups design is used wherein 230 child psychologists completed surveys regarding diagnostic practice. Psychologists differ in adherence with Diagnostic and Statistical Manual of Mental Disorders (4th ed., text revision) diagnostic criteria, use of clinical interviews, and type of behavior observation. Only 15% of psychologists report using multiple methods consistent with recommended standards of best practice. Differences between groups of psychologists indicate that the diagnosis of ADHD in children is influenced by the type of psychologist conducting the evaluation and the setting in which the evaluation is conducted. PMID- 16371664 TI - Perceptions of ADHD in China and the United States: a preliminary study. AB - College students (n = 226) and teachers (n = 328) in the United States and China completed a 55-item questionnaire examining perceptions of attention deficit/hyperactivity disorder (ADHD) in the two countries. Although a factor analysis yielded somewhat similar structures for the construct of ADHD, many differences emerged as well, with Chinese participants somewhat more attuned to hyperactivity than inattention. Furthermore, presented with a list of potential perceptions of ADHD, there were significant differences by culture in agreement with most of those statements. Thus, it appears that the concept of ADHD may be similar between the cultures, but the many differences warrant further exploration before ADHD is considered to be equivalent in the two countries. PMID- 16371665 TI - The role of treatment acceptability in the initiation of treatment for ADHD. AB - Approximately 50% of families of children with ADHD fail to pursue, or adhere to, recommended treatments. The present study examines parent ratings of the acceptability of pharmacological and behavioral treatments for ADHD and the relationships between these ratings and subsequent pursuit of treatment. Fifty five families whose children received an evaluation for ADHD completed questionnaires and were contacted 3 to 4 months later to assess their pursuit of treatment. Consistent with previous research, parents rated behavior therapy as more acceptable than medication. Parent ratings of medication acceptability significantly predict pursuit of pharmacological treatment, whereas ratings of the acceptability of behavior therapy do not predict pursuit of this treatment. Preliminary analyses found that Caucasian parents' ratings of medication are significantly higher than those of non-Caucasian parents. Furthermore, Caucasian families were more likely to pursue a recommendation for pharmacological treatment than non-Caucasian families. The clinical and research implications of these results are considered. PMID- 16371666 TI - Quality of life of men and women with borderline intelligence and attention deficit disorders living in community residences: a comparative study. AB - The research studies the quality of life (QOL) of 127 men and women diagnosed as having a borderline IQ and ADHD living in two major residential programs of the Sharon region in central Israel in respect to their personal, disability, and social ecological variables. Core findings indicate that men and women differ significantly according to their educational background. Among those who studied in a regular educational environment, the main predictor of men's QOL is their monthly income, whereas the main predictor for women is their level of attention deficit symptoms. The only predictor of QOL among men who studied in a special education program is the quality of contact with their mothers, whereas no significant predictor is observed among women. PMID- 16371667 TI - Comparison of measured and estimated cognitive ability in older adolescents with and without ADHD. AB - Premorbid intellectual function estimation is a crucial part of patient evaluation following a traumatic brain injury (TBI), especially in individuals with ADHD who are at higher risk for TBI compared to their non-ADHD peers. This study investigates the value of using regression-based estimates of intelligence for concurrently predicting measured intelligence in a sample of older adolescents with and without a childhood history of ADHD. Correlations between measured and estimated intelligence are highly significant in the full sample and in the individual groups. Adding reading performance to the regression equation increases the accounted-for variance in both groups. Results indicate that regression equations based on demographic characteristics constitute a valid method for estimating premorbid functioning in adolescents with ADHD and that they can play an essential role in the assessment of individuals with ADHD who sustain TBI, especially when measures of word reading are used to augment demographic estimates. PMID- 16371672 TI - Implementation of a comprehensive schoolwide behavioral intervention: The ABC program. AB - The Academic and Behavioral Competencies (ABC) Program, a schoolwide program to reduce classroom disruption and encourage rule following, academic task completion, and homework completion, is described. The program was initially developed and implemented in an elementary school with a high-risk population. Data from teachers, parents, and children indicate high levels of satisfaction with the program. In addition, unobtrusive measures of program impact, reported as reductions in referrals to the principal's office, suspensions, and increases in homework completion rates relative to the year prior to implementation of the program, suggest a preliminary positive impact of the program. A replication is reported for another school district, with teacher evaluations of satisfaction and effectiveness reported, supporting the flexibility and adaptability of the program. Although the present article does not constitute a systematic evaluation of the ABC Program, it presents preliminary data on the process of implementation and stakeholder satisfaction. PMID- 16371673 TI - School-based mental health programming for children with inattentive and disruptive behavior problems: first-year treatment outcome. AB - This article examines the effectiveness of an evidence-based behavioral treatment package for children with inattentive and disruptive behavior problems when delivered in the context of a school-based mental health program. Child symptomatology and functioning are assessed in a treatment group (n = 30) and a waitlist control group (n = 12) across multiple time points (fall, winter, and spring). Treatment includes a daily report card procedure, year-long teacher consultation, and parenting sessions. According to the parent report, treated children show marked reductions in hyperactive and impulsive, oppositional or defiant and aggressive behavior, and marked improvement in peer relationships. Teachers observe treatment-related group differences in inattention, academic functioning, and the student-teacher relationship. Feasibility and acceptability data have implications for transporting evidence-based treatments to community settings and for integrating mental health services into the culture of the school community. PMID- 16371674 TI - A laboratory school comparison of mixed amphetamine salts extended release (Adderall XR) and atomoxetine (Strattera) in school-aged children with attention deficit/hyperactivity disorder. AB - Mixed amphetamine salts extended release (MAS XR; Adderall XR) and atomoxetine (Strattera) were compared in children 6 to 12 years old with attention deficit/hyperactivity disorder (ADHD) combined or hyperactive/impulsive type in a randomized, double-blind, multicenter, parallel-group, forced-dose-escalation laboratory school study. Primary efficacy measure was the SKAMP (Swanson, Kotkin, Agler, M-Flynn, and Pelham) behavioral rating scale. Changes in mean SKAMP deportment scores from baseline were significantly greater for MAS XR (n = 102) than for atomoxetine (n = 101) overall (-0.56 and -0.13, respectively; p < .0001) and at each week (p < .001). Adverse events were similar for both treatment groups. The extended time course of action and greater therapeutic efficacy of MAS XR suggests that it is more effective than atomoxetine in children with ADHD. PMID- 16371675 TI - The relative effects of classwide peer tutoring and peer coaching on the positive social behaviors of children with ADHD. AB - This study investigates the effects of Classwide Peer Tutoring (CWPT) and peer coaching on the peer social behaviors of children with ADHD. A single-subject, multiple-baseline design is used with three elementary-school students in Grades 3 and 4. Following a baseline period, CWPT is implemented in each student's classroom. During the second intervention phase, CWPT is continued and peer coaching is added. Peer social behaviors are observed in both academic and social settings, with a primary focus on intervention effects on the latter setting. Results suggest that students participating in CWPT are actively and positively engaged with their peers while carrying out the CWPT program in the academic setting. However, when only CWPT is implemented, increases in positive peer social behaviors are not observed in social settings. The addition of peer coaching results in enhanced social behaviors during recess and lunch. PMID- 16371676 TI - The effects of computer-assisted instruction on the mathematics performance and classroom behavior of children with ADHD. AB - The present study examines the effects of computer-assisted instruction (CAI) on the mathematics performance and classroom behavior of three second-through fourth grade students with ADHD. A controlled case study is used to evaluate the effects of the computer software on participants' mathematics performance and on-task behavior. Participants' mathematics achievement improve and their on-task behavior increase during the CAI sessions relative to independent seatwork conditions. In addition, students and teachers consider CAI to be an acceptable intervention for some students with ADHD who are having difficulty with mathematics. Implications of these results for practice and research are discussed. PMID- 16371677 TI - The Behavior Education Support and Treatment (BEST) school intervention program: pilot project data examining schoolwide, targeted-school, and targeted-home approaches. AB - As part of a pilot project, four elementary schools were randomly assigned to receive one of four interventions: (a) a schoolwide intervention that incorporated universal and targeted treatment, (b) a targeted-school intervention delivered to individual students in regular and special education classrooms, (c) a targeted-home intervention delivered in home and regular classroom settings, and (d) a control condition that did not receive a designated intervention. Results showed that the behavior of disruptive children in all schools improved during the course of the year, with some evidence that interventions provided complementary effects. These findings support the continued use of behavioral interventions in elementary schools and argue for interventions that combine different methods of delivering interventions. PMID- 16371678 TI - The effect of conscious control on handwriting in children with attention deficit hyperactivity disorder. AB - Two experiments were performed regarding the effect of conscious control on handwriting fluency in healthy adults and ADHD children. First, 26 healthy students were asked to write a sentence under different conditions. The results indicate that automated handwriting movements are independent from visual feedback. Second, the writing performance of 12 children with ADHD was examined on their usual medication with methylphenidate and under placebo. In comparison to placebo, medication with methylphenidate resulted in a reduced fluency of handwriting. Automated handwriting movements could be elicited in children with ADHD on medication. The results suggest that both visual and mental control of handwriting movements affect the automation of handwriting movements. Furthermore, a simple training procedure was designed and performed in a case study of a boy with ADHD. PMID- 16371679 TI - Development of a school-wide behavior program in a public middle school: an illustration of deployment-focused intervention development, stage 1. AB - School-wide behavior management systems can improve academic performance and behavior in middle schools, and they should have positive effects on students with ADHD. Unfortunately, evidence-based, school-wide behavior management systems have not been widely adopted because of problems with feasibility, acceptability, and sustainability. The Deployment-Focused Model of Intervention Development and Testing has been proposed as a promising method of bridging the gap between research and practice settings. A key aspect of the model is to involve the persons most likely to deliver the service (e.g., teachers) in the intervention development process from the very beginning. To illustrate this process, the authors describe the planning and implementation of a school-wide program designed to improve behavior in a public middle school. PMID- 16371680 TI - Development of a school-based treatment program for middle school youth with ADHD. AB - The authors conduct an evaluation of a middle school-based treatment program for youth with ADHD during early stages of treatment development. The studies focus on interpreting outcome trends in preliminary data and identifying assessment issues that will be important to consider when conducting a clinical trial. Parent reports indicate that the majority of students benefit from improvements in academic, social, and overall functioning. Although teachers report beneficial effects for the majority of the participants in the program, there is little agreement about individual students. Measurement problems are associated with understanding normal change during a school year, accounting for normal behavior changes in December and May, and considerable disagreement between teachers. Suggestions to guide future work in this area are provided. PMID- 16371681 TI - Intervening to improve communication between parents, teachers, and primary care providers of children with ADHD or at high risk for ADHD. AB - This study examines interventions designed to improve communication between individuals who take care of children with ADHD. A teacher rating of 6,171 elementary school children identifies 1,573 children with ADHD or with high risk for ADHD. Parent interviews and information from teachers are collected on 243 children who are randomized into treatment and control conditions and followed for 39 months. The interventions consisted of group workshops and single one-on one tutorials with parents, teachers, and providers about the evaluation and treatment of ADHD that stressed the need for communication between the three parties. There are few significant effects on communication that are short lived. Results suggest that the interventions are insufficient to cause significant increase in communication. Future attempts to improve parent-teacher-provider coordination should be continuing rather than single-session interventions. PMID- 16371683 TI - Knowledge of zoology can help to clarify problems in epidemiology. PMID- 16371684 TI - Fox rabies in Germany - an update. AB - In comparison with conventional methods of wildlife rabies control, oral rabies vaccination of foxes (ORV) is without doubt the most (cost-) effective method in wildlife rabies control. As a result of ORV, several European countries have become rabies-free. Although rabies had been eliminated from much of Germany, there still exists a residual rabies focus in the border triangle of Hesse, Baden Wurttemberg and Rhineland Palatinate. Corrective actions have been initiated to eliminate this last remaining rabies hotspot in Germany. PMID- 16371685 TI - Rabies surveillance, trends in animal rabies and human post-exposure treatment in Poland, 1990 -2004. AB - This paper describes recent changes in the epizootical and epidemiological situation of rabies in Poland. Analysis of routine surveillance data on animal cases and human post-exposure treatment was performed in order to examine the impact of introduction of cell culture vaccine for human use and the implementation of the fox immunisation programme. The success of the immunisation programme for wild animals has become evident during the past 3 years, as a 9 fold decrease in animal rabies cases has been observed. To date, however, the downward trend in animal rabies cases has had no effect on the frequency of administration of the post-exposure treatment for humans. Moreover, two cases of locally acquired human rabies have occurred in patients who did not receive post exposure vaccination. These cases prove that rabies should be still considered a public health concern in Poland. PMID- 16371686 TI - A human case of travel-related rabies in Austria, September 2004. AB - A young male Austrian tourist, aged 23 years and unvaccinated against rabies, was bitten by a dog in Morocco in July 2004. One month later he was hospitalised in Ceuta with symptoms compatible with rabies. He died on 23 September in an Austrian hospital after a diagnosis of rabies was confirmed by FAT, IHC and RT PCR (including sequencing) of the neck skin and the RT-PCR (including sequencing) of the pharyngeal swab. This Austrian case of laboratory confirmed rabies highlights the urgent need for reinforcement of the international recommendations for travel vaccinations. PMID- 16371687 TI - An imported case of canine rabies in Aquitaine: investigation and management of the contacts at risk, August 2004-March 2005. AB - In August 2004, a case of rabies was diagnosed in a puppy that had been illegally imported from Morocco to Bordeaux (France). Because a great number of people and animals were thought to have come into contact with the puppy, extensive tracing measures were implemented, and an international alert was launched to trace and treat the contacts at risk. One hundred and eighty seven people received post exposure treatment, eight of whom also received serovaccination, and 57 animals known to have been exposed to the puppy were tested. Six months after the death of the rabid animal, none of the people treated showed any signs of rabies, nor was any secondary animal case reported. The management of this crisis highlights the importance of the role of a rapid alert system at European level. Strict application of sanitary control regulations is essential for animals introduced into EU countries, and all necessary information must be made available to EU residents travelling to rabies enzootic areas. PMID- 16371688 TI - Fox rabies in France. AB - Fox rabies was first recorded in France in March 1968, and remained a problem until 1998. In the course of the first two decades and despite the control measures applied, rabies expanded both in terms of the enzootic surface area and number of cases. The measures applied consisted of actions aimed at reducing fox population density, the mandatory vaccination of domestic carnivores in the officially infected areas, and use of human prophylaxis. Following the large scale implementation of oral vaccination of foxes, starting 1989-1990, the rabies front was pushed back and yearly incidence decreased until rabies was eliminated at the end of 1998. The comparison of results obtained during both periods of applying various strategies is spectacular. France remains exposed to the risk from bat rabies on one hand, and from accidental cases of canine rabies imported from enzootic countries, on the other. PMID- 16371689 TI - Public health concerns in bat rabies across Europe. AB - Rabies due to two independent and different genotypes of lyssaviruses - European bat lyssaviruses (EBLV) type 1 and type 2 - is present in many European countries. Infection is usually seen in bats, the primary reservoirs of the viruses but a few spillover infections have been seen in three other species: stone martens, sheep and humans. Spillover infections (with the exception of the two human cases) were EBLV-1 only. No EBLV-2 spillover cases have been reported in terrestrial animals. The disease is fatal in humans and has been described in Europe following a bat bite. We have studied in the available literature EBLV rabies cases across Europe in bats and humans, and have also carried out an analysis of recommendations for rabies prevention and treatments in humans. Rabies pre-exposure vaccination and post-exposure treatment is recommended for occupationally exposed persons. Some European countries have already adopted recommendations through specific protocols. Treatment of international travellers after bat bites is also recommended. The promoting of research programmes on bat rabies in Europe is underway. Bats are listed as protected species across Europe. PMID- 16371690 TI - Rabies in Europe in 2005. AB - Rabies is still present in Europe in 2005. Its incidence in humans remains limited (fewer than 5 human cases per year) through the application of strict prophylactic measures (anti-rabies treatment) and by means of veterinary rabies control measures in the domesticated and wild animal populations. The main indigenous animal reservoirs are: the dog in eastern European countries and on the borders with the Middle East; the fox in central and eastern Europe; the racoon dog in northeastern Europe; and the insectivorous bat throughout the entire territory. Finally, each year, cases of animals with rabies imported from enzootic areas are reported, showing the permeability of borders and traveller's lack of consideration of the rabies risk. These importations constantly threaten the rabies-free status of terrestrial animals in western European countries and complicate the therapeutic decisions taken by physicians in the absence of information regarding the biting animal. PMID- 16371691 TI - Rabies remains a 'neglected disease'. PMID- 16371692 TI - Legionnaires' disease in Europe 2003-2004. AB - Once a year, countries that collaborate in the European Surveillance Scheme for Travel Associated Legionnaires' Disease (EWGLINET) are requested to submit a dataset that provides epidemiological and microbiological information on cases of legionnaires' disease (nosocomial (hospital-acquired), community and travel related) detected in their country for that year. This paper presents the data collected for 2003 and 2004. For this period, 9166 cases were reported to the dataset by 35 countries, of which 941 cases were associated with outbreaks. Fourteen countries reported a total of 218 detected outbreaks. National infection rates varied between countries from 28.7 to less than one case per million population. This information is valuable in that it allows countries to assess the effectiveness of their national surveillance schemes in detecting cases. Over the two year period, 748 cases were reported to have died, giving a case fatality rate of 8.2%. The lack of detailed epidemiological information on deaths from legionnaires' disease is highlighted. The establishment of the European Centre for Disease Prevention and Control is seen as an opportunity to develop European collaborations more fully, and to increase further the protection of Europeans from outbreaks of legionnaires' disease. PMID- 16371693 TI - Epidemiology of invasive meningococcal disease in France in 2003. AB - National surveillance of invasive meningococcal disease (IMD) is based on mandatory reporting. The case definition for surveillance notification was changed in mid-2002 to include cases without microbiological confirmation. The IMD alert detection system was enhanced in 2003 with daily reporting and weekly analysis by district, serogroup, and age. Evaluation of the exhaustivity of the surveillance with capture-recapture analysis allowed correcting for underreporting. In 2003, 803 cases were reported. After correction for under reporting, the estimated incidence was 1.78 / 100,000. After excluding 'new' cases reported with new definition criteria, the 2002-2003 increase was 4%. Incidence decreased with age, with the highest values in infants less than 1 year (20/100,000), children aged between 1 and 2 years (11/100,000) and in teenagers of 17 years old(7/100,000). The overall case fatality rate was 12%. Fifty nine per cent of cases were due to serogroup B, 32% to C, 5% to W135, and 4% to Y and non-groupable meningococci. Patients with purpura fulminans treated with intravenous antibiotics before admission to hospital were shown to have lower fatality rates than those not treated. In 2001-2003, 5 situations required particular attention: two clusters of serogroup B IMD had set off mass prophylaxis, one outbreak due to a specific B IMD clonal complex with high case fatality rate, and two districts crossed the alert threshold for serogroup C IMD, 2/100,000, and mass vaccination was recommended. PMID- 16371694 TI - Ascertainment of meningococcal disease in Europe. AB - Meningococcal disease surveillance in most countries is based upon a combination of statutory notification systems and laboratory reporting, both of which are recognised to underestimate the true burden of disease. The incidence of meningococcal disease varies throughout Europe, and although there are many reasons for this, it is important to quantify the degree of under-ascertainment in order to validate international comparisons. Here, we review the literature on the ascertainment of meningococcal disease in Europe and the available methods for estimating the degree of under-reporting. We found that the sensitivity of surveillance varies between countries and over time, with estimates ranging from 40% to 96%. We identified five methods suitable for conducting ascertainment studies, from simple comparative studies to more complicated capture-recapture and regression analyses. Studies of ascertainment may be used to identify weaknesses and biases in surveillance data, and facilitate the improvement of these systems. These findings are relevant to the surveillance of other infectious diseases, particularly those with lower mortality and a lower public profile than meningococcal disease, for which ascertainment may be worse. PMID- 16371695 TI - 'Did you have flu last week?' A telephone survey to estimate a point prevalence of influenza in the Swedish population. AB - Sentinel surveillance usually underestimates the true burden of influenza in a population, as individuals must present to medical establishments to be included in the surveillance system. We carried out a telephone survey to estimate the national burden of influenza in the Swedish population for one week during the 2004/05 influenza season. Fixed-line telephone numbers were randomly sampled and households interviewed concerning influenza illness between 14-20 February 2005 (Week 7 of 2005). Questions regarding seasonal influenza vaccination status, symptoms and the impact of illness on daily life were also included. A self defined influenza prevalence of 7.7% in week 7 of 2005 was estimated. On applying a case definition of 'cough and fever and muscle pain' for influenza like illness, the prevalence decreased to 3.6%. The survey provided insight into the burden of illness in the population further to that estimated through the sentinel surveillance system. PMID- 16371696 TI - Human-to-human transmission of avian influenza A/H7N7, The Netherlands, 2003. AB - An outbreak of highly pathogenic avian influenza A virus subtype H7N7 began in poultry farms in the Netherlands in 2003. Virus infection was detected by RT-PCR in 86 poultry workers and three household contacts of PCR-positive poultry workers, mainly associated with conjunctivitis. To determine the magnitude of and risk factors for human-to-human transmission of influenza A/H7N7 in the Netherlands, a retrospective cohort study among household members of infected poultry workers was undertaken. In total, 33 (58.9%) of 56 (among 62) participants who provided blood samples had positive H7 serology, using single convalescent serum samples obtained at least 3 weeks after onset of symptoms of the index case. Eight household members (12.9%) reported symptoms (conjunctivitis and/or ILI), of which four of five (80.0%) tested seropositive. On univariate analysis, significant risk factors for seropositivity included having at least two toilets, a pet bird, and using cloth handkerchiefs. It was not possible to obtain a stable model for binomial regression for the outcome of A/H7N7 infection. Further seroprevalence studies among contacts of asymptomatic H7 cases should be conducted. PMID- 16371697 TI - Avian and pandemic influenza--five questions for 2006. PMID- 16371698 TI - X-ray diffraction by a crystal in a permanent external electric field: electric field-induced structural response in alpha-GaPO4. AB - For the first time, site-selective distortion has been investigated for two different structural units in the ternary compound alpha-GaPO(4) under the influence of a permanent external electric field. Based on 54 measured reflection intensities, the electric-field-induced distortion of PO(4) and GaO(4) tetrahedra in alpha-GaPO(4) crystals is evaluated using a model of pseudoatomic displacements introduced recently [Gorfman, Tsirelson & Pietsch (2005). Acta Cryst. A61, 387-396]. A stronger variation of the P-O bond lengths in the PO(4) tetrahedron was found compared to the bonds in the GaO(4) tetrahedron. The different distortions of the tetrahedra owing to the electric field were analysed in terms of the valence charge density of alpha-GaPO(4) and its topological characteristics. The larger charge of the P pseudoatom compared to the Ga atom was recognized as the main reason for the higher sensitivity of the PO(4) tetrahedron to a permanent external electric field. PMID- 16371699 TI - Reciprocal-space mapping of epitaxic thin films with crystallite size and shape polydispersity. AB - A development is presented that allows the simulation of reciprocal-space maps (RSMs) of epitaxic thin films exhibiting fluctuations in the size and shape of the crystalline domains over which diffraction is coherent (crystallites). Three different crystallite shapes are studied, namely parallelepipeds, trigonal prisms and hexagonal prisms. For each shape, two cases are considered. Firstly, the overall size is allowed to vary but with a fixed thickness/width ratio. Secondly, the thickness and width are allowed to vary independently. The calculations are performed assuming three different size probability density functions: the normal distribution, the lognormal distribution and a general histogram distribution. In all cases considered, the computation of the RSM only requires a two-dimensional Fourier integral and the integrand has a simple analytical expression, i.e. there is no significant increase in computing times by taking size and shape fluctuations into account. The approach presented is compatible with most lattice disorder models (dislocations, inclusions, mosaicity, ...) and allows a straightforward account of the instrumental resolution. The applicability of the model is illustrated with the case of an yttria-stabilized zirconia film grown on sapphire. PMID- 16371700 TI - Groupoid of orientational variants. AB - Daughter crystals in orientation relationship with a parent crystal are called variants. They can be created by a structural phase transition (Landau or reconstructive), by twinning or by precipitation. Internal and external classes of transformations defined from the point groups of the parent and daughter phases and from a transformation matrix allow the orientations of the distinct variants to be determined. These are algebraically identified with left cosets and their number is given by the Lagrange formula. A simple equation links the numbers of variants of the direct and inverse transitions. The equivalence classes on the transformations between variants are isomorphic to the double cosets (operators) and their number is given by the Burnside formula. The orientational variants and the operators constitute a groupoid whose composition table acts as a crystallographic signature of the transition. A general method that determines if two daughter variants can be inherited from more than one parent crystal is also described. A computer program has been written to calculate all these properties for any structural transition; some results are given for Burgers transitions and for martensitic transitions in steels. The complexity, irreversibility and entropy of fractal systems constituted by orientational variants generated by thermal cycling are briefly discussed. PMID- 16371702 TI - Radiation driven collapse of protein crystals. AB - During coherent X-ray diffraction measurements on crystals of ferritin at room temperature using monochromatic undulator radiation from the Advanced Photon Source, a sudden lattice contraction was observed following a characteristic latent period and ultimately leading to the collapse of the crystal. The progression of this collapse is analysed using a two-state Hendricks-Teller model. It reveals that 55% of the layers collapse by 1.6% before the crystal completely stops diffracting. PMID- 16371703 TI - The new diffractometer for surface X-ray diffraction at beamline BL9 of DELTA. AB - The experimental endstation of the hard X-ray beamline BL9 of the Dortmund Electron Accelerator is equipped with a Huber six-circle diffractometer. It is dedicated to grazing-incidence X-ray diffraction and X-ray reflectivity experiments on solid surfaces and thin films as well as to powder diffraction measurements. A new set-up for grazing-incidence X-ray scattering of liquids has been built up using a silicon mirror to reflect the incident X-ray to the liquid surface at angles of incidence around the critical angle of total reflection of the sample. X-ray reflectivity measurements of a polymer film and grazing incidence X-ray diffraction measurements of an epitaxically grown Gd40Y60 film, an oxidized surface of Fe-15at.%Al alloy and aqueous salt solutions are presented and discussed. PMID- 16371704 TI - High-temperature and high-pressure in situ SCC device for synchrotron radiation diffraction experiments and application using an austenitic stainless steel. AB - Suppressing the stress corrosion cracking (SCC) by reducing the carbon content in austenitic stainless steels is apparently not effective on core shrouds used in boiling water reactors in Japan: trans-granular cracking was found in the shrouds. To clarify the mechanism of the cracking, in situ stress measurements on specimens under stretched conditions in hot water have been attempted in the present study. An in situ device for diffraction measurements at synchrotron radiation facilities has been developed, and in situ experiments have been carried out at SPring-8. The SUS316L steel specimen was solution heat-treated, surface-ground and then placed in the in situ device. Sapphire windows were used for the light path in the device. A sufficient diffracted beam intensity was obtained through two sapphire windows and water. The side-inclination method was used for measuring the stress exerted on the specimen. A 2theta-sin2psi plot showed that a tensile stress was induced. The measured stress value is considered to be the summation of stresses owing to pre-straining, in situ loading and residual stress owing to surface grinding. PMID- 16371705 TI - Quantitative local structure refinement from XANES: multi-dimensional interpolation approach. AB - A new method to determine local structure in terms of a few structural parameters is proposed and realised in FitIt software. It is based on fitting of X-ray absorption near-edge structure (XANES) spectra using the combination of full multiple-scattering calculations, and multi-dimensional interpolation of spectra as a function of structural parameters. The procedure is divided into two steps: the construction of an interpolation polynomial, and the fitting of experimental spectra using the interpolation polynomial. During the construction of the polynomial, multiple-scattering calculations for certain sets of structural parameters are needed. The strategy for the selection of the most important expansion terms and corresponding sets of structural parameters is proposed. Fitting of the spectrum using multi-dimensional interpolation is very fast (a few seconds) because multiple-scattering calculations are unnecessary during this step. Also, this approach allows the development of a visual interface with the possibility of seeing the spectrum that corresponds to any set of structural parameters immediately. Thus, using a very limited number of multiple-scattering calculations, which are most time-consuming, it is possible to fit XANES. The interpolation polynomial construction procedure for three model molecules, FeS4, FeO6 and Ni(CN)4, is demonstrated. An additional test has been performed for the latter most-complex example to check the assumption that a minimum of discrepancy between theoretical and experimental spectra corresponds only to the correct structure of the complex. A comparison with another XANES fitting software, MXAN, is given. PMID- 16371706 TI - The Structural Biology Center 19ID undulator beamline: facility specifications and protein crystallographic results. AB - The 19ID undulator beamline of the Structure Biology Center has been designed and built to take full advantage of the high flux, brilliance and quality of X-ray beams delivered by the Advanced Photon Source. The beamline optics are capable of delivering monochromatic X-rays with photon energies from 3.5 to 20 keV (3.5-0.6 A wavelength) with fluxes up to 8-18 x 10(12) photons s(-1) (depending on photon energy) onto cryogenically cooled crystal samples. The size of the beam (full width at half-maximum) at the sample position can be varied from 2.2 mm x 1.0 mm (horizontal x vertical, unfocused) to 0.083 mm x 0.020 mm in its fully focused configuration. Specimen-to-detector distances of between 100 mm and 1500 mm can be used. The high flexibility, inherent in the design of the optics, coupled with a kappa-geometry goniometer and beamline control software allows optimal strategies to be adopted in protein crystallographic experiments, thus maximizing the chances of their success. A large-area mosaic 3 x 3 CCD detector allows high quality diffraction data to be measured rapidly to the crystal diffraction limits. The beamline layout and the X-ray optical and endstation components are described in detail, and the results of representative crystallographic experiments are presented. PMID- 16371707 TI - X-ray powder diffraction beamline at D10B of LNLS: application to the Ba2FeReO6 double perovskite. AB - A new beamline, fully dedicated to X-ray powder diffraction (XPD) measurements, has been installed after the exit port B of the bending magnet D10 at the Brazilian Synchrotron Light Laboratory (LNLS) and commissioned. The technical characteristics of the beamline are described and some performance indicators are listed, such as the incoming photon flux and the angular/energy resolutions obtainable under typical experimental conditions. The results of a Rietveld refinement for a standard sample of Y2O3 using high-resolution data are shown. The refined parameters match those found in the literature, within experimental error. High-resolution XPD measurements on Ba2FeReO6 demonstrate a slight departure from the ideal cubic double-perovskite structure at low temperatures, not detected by previous powder diffraction experiments. The onset of the structural transition coincides with the ferrimagnetic ordering temperature, Tc approximately equal to 315 K. Subtle structural features, such as those reported here for Ba2FeReO6, as well as the determination and/or refinement of complex crystal structures in polycrystalline samples are ideal candidate problems to be investigated on this beamline. PMID- 16371708 TI - A wide-aperture dynamically focusing sagittal monochromator for X-ray spectroscopy and diffraction. AB - A scanning dynamically focusing sagittal X-ray monochromator accepting 7 mrad of the fan from a 6 T wiggler is in routine use on beamline 16.5 (ultra-dilute spectroscopy) of the SRS at CCLRC Daresbury Laboratory, UK. The energy range covered is 7-27 keV, with a horizontal spot size of <1.1 mm FWHM. Measured monochromatic flux from a Si 220 crystal pair is 1 x 10(11) photons s(-1) (100 mA)(-1) at 9 keV. This level of flux, usually associated with an insertion device on a third-generation source, permits collection of EXAFS data on concentrations at or below 10 ppm. PMID- 16371709 TI - Feedback system of a liquid-nitrogen-cooled double-crystal monochromator: design and performances. AB - A new set-up is reported of an indirect cryogenic cooling system for a double crystal monochromator which runs on the BM30b/FAME beamline at the ESRF (Grenoble, France). This device has been conceived to limit the vibrations on the first diffracting crystal and to maintain it at a constant temperature. These points are crucial for maximizing the beamline stability. Moreover, the relative angular position of the second crystal can be dynamically adjusted by a piezoelectric transducer coupled with a feedback system in order to always be at the maximum of the rocking curve during an X-ray absorption spectroscopy scan. The temperature is stabilized to an accuracy of 0.01 degrees , with two principal consequences. The energy resolution is close to the theoretical value [DeltaE/E = 5.6 x 10(-6) for Si(220)] and the precision of the energy positioning is extremely good even if the power load changes. A feedback mechanism allows a permanent and automatic optimization of the angle between the two crystals of the monochromator. The intensity of the monochromatic beam remains optimized (i) when the intensity of the electron beam decreases in the storage ring and (ii) during an energy scan. PMID- 16371710 TI - Coherent propagation of white X-rays in a planar waveguide. AB - The far-field diffraction pattern of a front-coupled planar waveguide supporting two guided modes has been measured using a white X-ray beam. Interference of the guided modes leads to a characteristic variation of the far-field diffraction pattern for different photon energies. The experiment verifies the predicted properties of the guided modes, shows that these modes superpose coherently, and demonstrates that the electromagnetic field downstream of the waveguide is significantly different from that expected for a hypothetical small slit of the same size. PMID- 16371711 TI - Single-bounce monocapillaries for focusing synchrotron radiation: modeling, measurements and theoretical limits. AB - Single-bounce hollow glass capillaries with ellipsoidal shapes have been used at the Cornell High Energy Synchrotron Source recently for various microbeam experiments, with focal spot sizes from 12 to 23 microm, divergences from 2 to 8 mrad, intensities up to 450 times the intensities of incident X-rays, and working distances up to 55 mm. Simple formulae are developed in this paper to explain capillary performance given the X-ray source size, capillary dimensions and slope errors. Capillary length is optimized for best focusing performance. Capillary fabrication accuracy is reported and capillary X-ray tests confirm the focusing properties expected from formulae. The application of capillaries to third generation X-ray sources and future energy-recovery linac X-ray sources are discussed. PMID- 16371712 TI - Large-distance refocusing of a submicrometre beam from an X-ray waveguide. AB - Among the several available X-ray optics for synchrotron radiation producing micrometre and submicrometre beams with high intensity, the X-ray waveguide (WG) can provide the smallest hard X-ray beam in one direction. A drawback of this optics is that, owing to the divergence at the exit, a nanometre-sized spot on the sample can only be obtained if this is within a few micrometres of the WG exit. Another limitation is that in planar WGs the beam is compressed in only one direction. Here, using a dynamically bent elliptical Si/Pt mirror, the guided X ray beam has been refocused at approximately 1 m from the waveguide exit. The large working distance between the device and the submicrometre focus leaves some space for sample environment (vacuum chamber, furnace, cryostat, magnets, high pressure device etc.) and allows cross-coupled geometries with two WGs for efficient compression in two directions. PMID- 16371715 TI - [Wernicke encephalopathy during total parenteral nutrition due to acute necrotizing pancreatitis]. PMID- 16371716 TI - [Robotics and gastrointestinal surgery]. AB - Robotics are now being used in all surgical fields. Because of increased intra abdominal articulations while operating through small incisions, robotics are increasingly being used in a large number of visceral and solid organ operations including surgery on the gallbladder, esophagus, stomach, intestines, colon, and rectum as well as for the endocrine organs. As a specialty, robotics should continue to grow. As the robotic era invades the field of general surgeon, more and more complex procedures would be able to be approached through small incision. As technology catches up with our imagination, robotic instruments and 3D monitoring will become routine, and continue to improve patient care by providing surgeons with most precise, least traumatic ways of treating surgical disease. PMID- 16371717 TI - [Comparison of Helicobacter pylori eradication rate according to different PPI based triple therapy--omeprazole, rabeprazole, esomeprazole and lansoprazole--]. AB - BACKGROUND/AIMS: Helicobacter pylori (H. pylori) is an important cause of various gastrointestinal diseases. H. pylori eradication is essential for the cure and prevention of associated diseases. Nowdays, proton pump inhibitor (PPI)-based triple therapy is the standard eradication regimen. The aims of this study were to compare the H. pylori eradication rate of different PPI-based triple therapies and to find out the factors influencing the eradication rate. METHODS: From May 2002 through February 2004, H. pylori infected patients were treated with the eradication regimen based on one of the four PPIs (omeprazole, rabeprazole, esomeprazole and lansoprazole) for 1 or 2 weeks. After two weeks, drug compliance, adverse effects, and smoking history during the eradication therapy were obtained. The follow-up H. pylori test was performed 4 weeks after the completion of therapy. The data were analyzed by Chi-square test and multiple logistic regression analysis. RESULTS: Overall eradication rate was 83.5%. There was no significant difference in eradication rate among four PPIs (p=0.379). Odds ratio (OR) for omeprazole and rabeprazole was 1.15 (95% CI 0.50-2.68); for omeprazole and esomeprazole, OR 1.63 (95% CI 0.68-3.89); and for omeprazole and lansoprazole, OR 1.13 (95% CI 0.50-2.56). Smoking habit, site of ulcer, and the duration of therapy affected the eradication rate significantly. CONCLUSIONS: The efficacy of four different PPIs for H. pylori eradication is similar to each other. Smoking, site of ulcer, and the duration of treatment have significant effects on eradication rates. PMID- 16371718 TI - [Effects of TNF-alpha on angiogenesis in CT-26 mouse colon carcinoma homograft model]. AB - BACKGROUND/AIMS: Tumor necrosis factor-alpha (TNF-alpha) exerts its anti-tumor effect through direct cytotoxicity on tumor cells and damage to the tumor vasculature. However, its role in tumor angiogenesis is controversial. We evaluated the angiogenic effect of TNF-alpha on BALB/c mouse colon carcinoma homograft model. METHODS: Ten BALB/c mice were inoculated intraperitoneally with CT-26 mouse colon carcinoma cells. After a week, recombinant mouse TNF-alpha (2 microg/mL) were given four times on every other day to five animals and the same volume of phosphate buffered saline was given at the same interval to five animals as control. Harvested tumor tissues were stained by immunohistochemistry with CD31 and VEGF antibodies. Number of microvessels and VEGF expression were counted by light microscope. RESULTS: The mean microvessel counts per 200x field of TNF-alpha treated animals were 70.2+/-7.8 and those of nontreated animals were 83.8+/-8.3 (p<0.05). The VEGF score of both groups were 3. CONCLUSIONS: TNF-alpha treated animals showed decreased microvessel counts in tumor tissue but VEGF expression in both groups showed no difference. Therefore, TNF-alpha showed antiangiogenic effects on colon carcinoma homograft model. PMID- 16371719 TI - Predictive factors of recurrent bleeding in Mallory-Weiss syndrome. AB - BACKGROUND/AIMS: Although the majority of patients with Mallory-Weiss syndrome (MWS) have a benign course, MWS patients with recurrent bleeding have an unfavorable outcome and require intensive care. Therefore, this study was carried out to identify the risk factors for recurrent bleeding in MWS patients. METHODS: The medical records of patients with MWS between January 1999 and December 2003, were reviewed retrospectively. Demographics, initial clinical and laboratory parameters, and endoscopic findings of the patients with and without recurrent bleeding were compared and the potential risk factors predicting recurrent bleeding in MWS were evaluated. RESULTS: A total of one hundred and fifty-nine patients (22 women, 137 men, mean age 48.1 years old) were enrolled in the study. Recurrent bleeding was observed in 17 patients (10.7%). Those patients with recurrent bleeding showed higher frequency for the presence of shock at initial manifestation, combined liver cirrhosis and endoscopic findings of active bleeding, lower hemoglobin level and platelet count, higher amount of transfusions and epinephrine-mixed fluid injections, and longer hospital stay than those patients without recurrent bleeding. Significant risk factors predicting the recurrent bleeding in MWS were the presence of shock at initial manifestation (OR 3.71, 95% CI 1.07-14.90) and the evidence of active bleeding on endoscopic examination (OR 9.89, 95% CI 1.88-51.98) on multivariate analysis. CONCLUSIONS: Intensive care with close monitoring is required for the patients with shock on initial manifestation or with evidence of active bleeding on endoscopic examinations since these are independent risk factors predicting the recurrent bleeding in MWS patients. PMID- 16371720 TI - [Characteristics of colon cancer diagnosed in patients taking aspirin or warfarin]. AB - BACKGROUND/AIMS: Warfarin and aspirin are commonly used to prevent cardiovascular diseases. Aspirin was recently found to have chemopreventive effects on colon cancer and polyps by inhibiting cyclooxygenase-2. Therefore, we evaluated whether the symptoms of bleeding related with aspirin or warfarin could be a clue in early detection of colon cancer. We also assessed the effect of aspirin on the development of synchronous polyps. METHODS: A total of forty-one and 16 patients diagnosed as colon cancer, taking aspirin or warfarin respectively were enrolled. In addition, 171 patients with colon cancers were age and gender matched as a control group. We investigated the difference of clinical features and laboratory findings among three groups. RESULTS: The incidence of bleeding was 81.3% (warfarin), 53.7% (aspirin), 40.4% (control). Among three groups, location and size of cancer, number of lymph nodes involvement and stages were not different, but the number of patients in Duke stage D in warfarin group (n=1, 6.3%) were less than that of the control (n=44, 25.7%) (p=0.049). The extent of circumferential involvement by cancer was lower in aspirin group (67%) than in the control group (80%) (p=0.035). The percentage of patients with synchronous polyps and mean number of synchronous polyps in aspirin group (34.1%, 0.68, respectively) was lower than that of control group (53.6%, 1.69, respectively) (p=0.029, 0.008, respectively). CONCLUSIONS: Bleeding related with aspirin or warfarin usage had no effect on the early diagnosis of colon cancer. However, lower incidence of Duke stage D in warfarin group might be related to anti metastatic effect of warfarin. In addition, aspirin may have a role in suppressing the development of synchronous polyps. PMID- 16371721 TI - [The outcome of endoscopic treatment in bile duct injury after cholecystectomy]. AB - BACKGROUND/AIMS: Bile duct injury is the most serious complication of cholecystectomy. The aim of this study was to evaluate the outcome of endoscopic treatment in bile duct injury after cholecystectomy. METHODS: We reviewed the results of endoscopic treatments in the patients diagnosed as bile duct injury after cholecystectomy on cholangiographic examinations, retrospectively. Endoscopic treatment included insertion of nasobiliary drainage catheter or plastic stent after endoscopic sphicterotomy. RESULTS: A total of twenty-two patients (9 male, 13 female; median age of 59 years) with bile duct injury were included. Endoscopic treatment was successfully performed in 12 of 13 patients with bile leak only. In patients with both bile leak and stricture, endoscopic treatment was successful in 2 of 3 patients. In 6 patients with complete obstruction of bile duct, endoscopic treatment failed and surgical approach was needed. In our series, transpapillary endoscopic treatment was not successful when proximal bile duct above the injured site was not visualized by endoscopic retrograde cholangiopancreatography (ERCP) and surgery was performed in all cases. Overall success rate of endoscopic treatment in 22 patients with bile duct injury was 64% (14/22). There was no complication associated with endoscopic treatment. CONCLUSIONS: ERCP is useful for the treatment of bile leakage after cholecystectomy and can be used for the treatment prior to surgery. Surgical intervention is needed in case of endoscopic treatment failure. PMID- 16371722 TI - [A case of subphrenic abscess with ileal fistula caused by metastatic adenocarcinoma of unknown origin]. AB - Usual sources of subphrenic abscess with intestinal fistula are previous abdominal operation, inflammatory bowel disease and malignancy. Reported cases of intestinal fistula caused by adenocarcinoma were complicated by direct invasion. In this report, a 70-year-old male had a subphrenic abscess with intestinal fistula and the cause was a metastatic adenocarcinoma of unknown origin. As far as we know, this has not been reported previously in the literatures. The abscess went on chronic course for six months because intermittent administration of antibiotics modified its clinical presentation. The fistulous tract between the abscess and ileum was demonstrated by tubogram via the drainage catheter in abscess. The patient underwent surgical treatment because the cause of fistula was obscure. Invasion of the ileum by metastatic adenocarcinoma was diagnosed by the histologic examination of surgical specimen. Therefore, when a fistula develops without any apparent cause, there is a possibility of malignancy, and surgical approach must be considered. An early surgical approach will prevent the delay in treatment and reduce the mortality. PMID- 16371723 TI - Acute pancreatitis due to hypertriglyceridemia: report of 2 cases. AB - Hypertriglyceridemia (HTG) is a rare but well known cause of acute pancreatitis (AP), which can be a life- threatening complication if the degree of HTG is severe enough. It might be primary in origin or secondary to alcohol abuse, diabetes mellitus, pregnancy, or drugs. A serum triglyceride (TG) level of more than 1,000 to 2,000 mg/dL in patients with type I, IV, or V hyperlipidemia (Fredrickson's classification) is the identifiable risk factor. HTG-induced AP typically presents as an episode of AP or recurrent AP. The clinical course of HTG-induced AP is not different from other causes. Routine management of HTG induced AP should be similar to other causes. A thorough family history of lipid abnormalities should be obtained, and an attempt to identify secondary causes should be made. The mainstay of treatment includes dietary restriction of fatty meal and lipid-lowering medications (mainly fibric acid derivatives). Although there are limited experiences with plasmapheresis, lipid apheresis, heparinization and insulin application, these can support the treatment of HTG- induced AP. We report two cases of HTG-induced AP which were successfully treated by plasmapheresis. PMID- 16371724 TI - [A case of gastric metastasis of breast carcinoma resembling early gastric cancer]. AB - Many patients suffering from breast carcinoma have metastases at initial diagnosis. The common metastatic sites are skeleton, liver and lung. Metastases to stomach are rare and only three cases have been reported in Korea. The endoscopic features of gastric metastases from breast carcinoma can be divided into three main categories: diffuse infiltration, external compression, and localized tumor deposition with ulceration or with a polypoid mass. However, metastatic gastric lesions which resemble early gastric carcinoma are rare. Typically, gastric metastases are confined to submucosa and muscularis, so that mucosal biopsy specimens might be false-negative. We report a case of gastric metastasis from infiltrative lobular carcinoma of the breast in a 66-year-old woman who had undergone left mastectomy with postoperative radiotherapy 17 years earlier. Initial diagnosis was early gastric carcinoma, signet ring cell type on gastric biopsy findings. However, definitive diagnosis of metastatic breast cancer was confirmed after endoscopic mucosal resection of a presumed primary early gastric carcinoma. PMID- 16371725 TI - [A case of pancreaticoportal fistula associated with acute severe pancreatitis]. AB - Pancreatic fistulas are usually caused by the disruption of pancreatic duct. The majority of pancreatic fistulas are external fistulas and common causes of external and internal pancreatic fistulas are trauma and surgery. Internal pancreatic fistulas due to pancreatitis are rare. Internal pancreatic fistulas may communicate with peritoneal cavity, colon, small bowel, biliary system or pleural cavity. Among them, fistula between pancreatic duct and portal vein due to acute pancreatitis is rare. We report a case of 32-year-old male with fistula between pancreatic duct and portal vein as a complication of acute pancreatitis. Pancreaticoportal fistula was diagnosed by endoscopic retrograde cholangiopancreatography. He recovered after distal pancreatectomy with splenectomy and supportive care. PMID- 16371726 TI - [Aspirin, is it effective for prevention of colon cancer?]. PMID- 16371728 TI - Influence of preoperative chemotherapy on the risk of major hepatectomy for colorectal liver metastases. AB - OBJECTIVE: : To assess the effects of preoperative systemic chemotherapy on remnant liver parenchyma, liver function, and morbidity after major liver resection for colorectal liver metastases. BACKGROUND: : Many patients operated upon for colorectal cancer liver metastases receive previous chemotherapy. Whether systemic chemotherapy alters liver parenchyma in such way that it increases the risks of liver resection remains unclear. PATIENTS AND METHODS: : Among 214 patients who received a liver resection for colorectal liver metastases between 1998 and 2002 in a single institution, 67 who underwent a major liver resection under total hepatic vascular exclusion form the basis of this report. Forty-five patients operated upon after systemic chemotherapy were compared with 22 who did not receive any chemotherapy in the 6 months prior to resection. Postoperative mortality, morbidity, liver function tests, and pathology of the resected liver in the two groups were compared. RESULTS: : There was no postoperative mortality. Values of liver function tests on days 1, 3, 5, and 10 were similar in both groups. Morbidity rate was higher in the chemotherapy group (38% versus 13.5%, P = 0.03). Postoperative morbidity was correlated with the number of cycles of chemotherapy administered before surgery but not to the type of chemotherapy. Preoperative chemotherapy was significantly associated with sinusoidal dilatation, atrophy of hepatocytes, and/or hepatocytic necrosis (49% versus 25%, P = 0.005). CONCLUSION: : Prolonged neoadjuvant systemic chemotherapy alters liver parenchyma and increases morbidity after major resection under total hepatic vascular exclusion, but it does not increase operative mortality. This should be taken into consideration before deciding a major liver resection in patients who have received preoperative chemotherapy. PMID- 16371729 TI - CASH (Chemotherapy-Associated Steatohepatitis) costs. PMID- 16371730 TI - Increasing operating room efficiency through parallel processing. AB - OBJECTIVE: Because of rising costs and shrinking reimbursements, hospitals must continually find ways to improve efficiency and productivity. This study attempts to increase caseloads in ambulatory surgery operating rooms while maintaining patient satisfaction and safety. SUMMARY BACKGROUND DATA: In most hospitals, patients move through their operative day in a linear fashion, starting at registration and finishing in the recovery room. Given this pattern, only 1 patient may occupy the efforts of the operating room team at a time. By processing patients in a parallel fashion, operating room efficiency and patient throughput are increased while costs remain stable. METHODS: Patients undergoing hernia repairs under local anesthesia with intravenous sedation were divided into a control group and an experimental group. Patients in the control group received their local anesthesia in the operating room at the start of the surgery. The experimental group patients received their local anesthesia in the induction room by the surgeon while the operating room was being cleaned and set up. RESULTS: While operative time for the control group and the experimental group were nearly identical, the turnover time and the induction time were significantly shorter for the experimental group. The cumulative reduction in time during the operative day was sufficient to allow the addition of new operative cases. CONCLUSIONS: This study demonstrates a system of increasing operating room efficiency by changing patient flow rather than simply working to streamline existing steps. This increase in efficiency is not associated with the expansion of hospital budgets or a decrease in patient safety or satisfaction. PMID- 16371731 TI - Assessing operating room efficiency and parallel processing. PMID- 16371732 TI - Laparoscopic versus open appendectomy in children: a meta-analysis. AB - OBJECTIVE: This study aims to use meta-analysis to compare laparoscopic and open appendectomy in a pediatric population. SUMMARY BACKGROUND DATA: Meta-analysis is a statistical tool that can be used to evaluate the literature in both qualitative and quantitative ways, accounting for variations in characteristics that can influence overall estimate of outcomes of interest. Meta-analysis of laparoscopic versus open appendectomy in a pediatric population has not previously been performed. METHODS: Comparative studies published between 1992 and 2004 of laparoscopic versus open appendectomy in children were included. Endpoints were postoperative pyrexia, ileus, wound infection, intra-abdominal abscess formation, operative time, and postoperative hospital stay. RESULTS: Twenty-three studies including 6477 children (43% laparoscopic, 57% open) were included. Wound infection was significantly reduced with laparoscopic versus open appendectomy (1.5% versus 5%; odds ratio [OR] = 0.45, 95% confidence interval [CI], 0.27-0.75), as was ileus (1.3% versus 2.8%; OR = 0.5, 95% CI, 0.29-0.86). Intra-abdominal abscess formation was more common following laparoscopic surgery, although this was not statistically significant. Subgroup analysis of randomized trials did not reveal significant difference between the 2 techniques in any of the 4 complications. Operative time was not significantly longer in the laparoscopic group, and postoperative stay was significantly shorter (weighted mean difference, -0.48; 95% CI, -0.65 to -0.31). Sensitivity analysis identified lowest heterogeneity when only randomized studies were considered, followed by prospective, recent, and finally large studies. CONCLUSIONS: The results of this meta-analysis suggest that laparoscopic appendectomy in children reduces complications. However, we also see the need for further high-quality randomized trials comparing the 2 techniques, matched not only for age and sex but also for obesity and severity of appendicitis. PMID- 16371733 TI - "Anatomic" right hepatic trisectionectomy (extended right hepatectomy) with caudate lobectomy for hilar cholangiocarcinoma. AB - BACKGROUND: The techniques of right hepatic trisectionectomy are now standardized in patients with hepatocellular or metastatic carcinoma, but not in those with hilar cholangiocarcinoma. METHODS: Under preoperative diagnosis of hilar cholangiocarcinoma, 8 patients underwent "anatomic" right hepatic trisectionectomy with en bloc resection of the caudate lobe and the extrahepatic bile duct, in which the bile ducts of the left lateral section were divided at the left side of the umbilical fissure following complete dissection of the umbilical plate. RESULTS: Liver resection was successfully performed, and all patients were discharged from the hospital in good condition, giving a mortality of 0%. All patients were histologically diagnosed as having cholangiocarcinoma. The proximal resection margins were cancer-negative in 7 patients and cancer positive in 1 patient. Four patients with multiple lymph node metastases died of cancer recurrence within 3 years after hepatectomy. One patient died of liver failure without recurrence 42 months after hepatectomy. The remaining 3 patients without lymph node metastasis are now alive after more than 5 years. CONCLUSIONS: Anatomic right hepatic trisectionectomy with caudate lobectomy can produce a longer proximal resection margin and can offer a better chance of long-term survival in some selected patients with advanced hilar cholangiocarcinoma. PMID- 16371734 TI - Early ductal decompression versus conservative management for gallstone pancreatitis with ampullary obstruction: a prospective randomized clinical trial. AB - OBJECTIVE: To compare the efficacy of endoscopic retrograde cholangiopancreatography +/- endoscopic sphincterotomy (ERCP +/- ES) versus traditional conservative management in early gallstone pancreatitis with persistent ampullary obstruction (GSP + AO). SUMMARY BACKGROUND DATA: The effectiveness of early ERCP +/- ES in this setting is controversial. METHODS: Sixty-one consecutive patients with GSP + AO within 48 hours from the onset of symptoms were randomized to receive either conservative treatment and selective ERCP +/- ES after 48 hours (control group, 31 patients) or initial conservative treatment and systematic ERCP +/- ES within 48 hours if obstruction persisted 24 hours or longer (study group, 30 patients). Patient outcome was compared in relation to treatment groups and to duration of obstruction. RESULTS: In the control group, 22 patients disobstructed spontaneously within 48 hours; 3 of the remaining 9 patients underwent ERCP +/- ES and none had impacted stones. In the study group, 16 patients disobstructed spontaneously and 14 underwent ERCP within 48 hours from the onset of symptoms; impacted stones were found and extracted by ES in 79% (11 of 14) of these. PATIENTS: There were no deaths in either group. Patients in the study group showed a shorter period of obstruction (P = 0.016) and a lower rate of immediate complications (P = 0.026) than controls. Patients with obstruction lasting < or =48 hours regardless of the treatment group had fewer immediate complications than those whose obstruction persisted longer (P < 0.001). CONCLUSIONS: This study shows that in patients with GSP + AO limiting the duration of obstruction to not longer than 48 hours by ERCP + ES decreased morbidity. PMID- 16371735 TI - Laparoscopic surgery may be associated with severe pain and high analgesia requirements in the immediate postoperative period. AB - OBJECTIVE: To assess the immediate (0-4 hours) postoperative pain level in patients after laparoscopy and laparotomy whose analgesic requirement in the Post Anesthesia Care Unit (PACU) exceeds standard morphine therapy. BACKGROUND DATA: Clinical observation has raised the suspicion that laparoscopic surgery may be associated with more intense immediate postoperative pain than expected. METHODS: This prospective study assessed the 24-hour pain intensity and analgesia requirements in patients who underwent similar abdominal surgery via laparoscopy or laparotomy under standardized general anesthesia and whose pain in the PACU was resistant to 120 microg/kg intravenous morphine. RESULTS: Of 145 sampled PACU patients, 67 were in pain (> or =6 of 10 VAS) within a 30-minute postoperative period. They were then given up to 4 intravenous boluses of 15 microg/kg morphine + 250 microg/kg ketamine. The pain VAS of 36 laparotomy patients was 4.14 +/- 2.14 (SD) and 1.39 +/- 0.55 at 10 and 120 minutes, respectively, after 1.33 +/- 0.59 doses of morphine + ketamine; the pain VAS of 31 laparoscopy patient was 6.06 +/- 1.75 and 2.81 +/- 1.14, respectively (P < 0.0005) following 2.0 +/- 0.53 doses (P = 0.0005). Diclofenac 75 mg intramuscular usage was similar (P = 0.43) between the groups up to 9 hours after surgery but was higher in the laparotomy group by 24 hours (P = 0.01). Pain scores at 24 hours after surgery were lower for the laparoscopy patients (3.01 +/- 0.87) compared with their laparotomy counterparts (4.45 +/- 0.98, P < 0.001). CONCLUSIONS: Among patients after abdominal surgery with severe immediate (0-4 hours) postoperative pain, laparoscopic patients are a significant (46%) proportion, and their pain is more intense, requiring more analgesics than painful patients (54%) do after laparotomy. By 24 hours, the former are in less pain than the latter. PMID- 16371737 TI - Efficacy of Nissen fundoplication versus medical therapy in the regression of low grade dysplasia in patients with Barrett esophagus: a prospective study. AB - OBJECTIVE: The aim of this study is to compare the effect of medical and surgical treatment on the history of patients with Barrett esophagus (BE) and histologic evidence of low-grade dysplasia (LGD). SUMMARY BACKGROUND DATA: BE is a complication of severe gastroesophageal reflux. It is considered a major risk factor for esophageal adenocarcinoma, which may develop through stages from nondysplastic metaplasia to dysplasia (LGD and high-grade dysplasia). Presently, there are no recommended therapeutic guidelines for patients with LGD. METHODS: Between 1998 through 2003, 6592 patients underwent upper endoscopy; 327 of 6592 (5%) patients had BE, and 35 of 327 (10.7%) had LGD. Nineteen patients with LGD were treated with high-dose proton pump inhibitors, and 16 patients underwent laparoscopic Nissen fundoplication. Endoscopic and histologic follow-up was available in all patients after 18 months. We used multiple logistic regression to examine the effect of the 2 treatments on regression of LGD. RESULTS: LGD was predominant in men (male-to-female ratio: 1.7:1). Mean age was 58 +/- 13.5 years. Sixty percent of patients had no endoscopic evidence of esophagitis. A regression from LGD to BE was observed in 12 of 19 (63.2%) patients in the medical group and in 15 of 16 (93.8%) patients in the surgical group (statistically significant difference). Differences between the 2 groups were statistically significant (P = 0.03). CONCLUSION: The results of our study suggest that surgical treatment may be more effective than medical therapy to modify the natural history of LGD in patients with BE, perhaps because it not only controls acid but also biliopancreatic reflux into the esophagus. PMID- 16371736 TI - The immunomodulatory effects of hypertonic saline resuscitation in patients sustaining traumatic hemorrhagic shock: a randomized, controlled, double-blinded trial. AB - OBJECTIVE: To investigate the potential immunologic and anti-inflammatory effects of hypertonic saline plus dextran (HSD) in hemorrhagic trauma patients. BACKGROUND: Unbalanced inflammation triggered by shock has been linked to multiorgan dysfunction (MOD) and death. In animal and cellular models, HSD alters the inflammatory response to shock, attenuating MOD and improving outcome. It remains untested whether HSD has similar effects in humans. METHODS: A single 250 mL dose of either HSD (7.5% NaCl, 6% dextran-70) or placebo (0.9% NaCl) was administered to adult blunt trauma patients in hemorrhagic shock. The primary outcome was to measure changes in immune/inflammatory markers, including neutrophil activation, monocyte subset redistribution, cytokine production, and neuroendocrine changes. Patient demographics, fluid requirements, organ dysfunction, infection, and death were recorded. RESULTS: A total of 27 patients were enrolled (13 HSD) with no significant differences in clinical measurements. Hyperosmolarity was modest and transient, whereas the immunologic/anti inflammatory effects persisted for 24 hours. HSD blunted neutrophil activation by abolishing shock-induced CD11b up-regulation and causing CD62L shedding. HSD altered the shock-induced monocyte redistribution pattern by reducing the drop in "classic" CD14 and the expansion of the "pro-inflammatory" CD14CD16 subsets. In parallel, HSD significantly reduced pro-inflammatory tumor necrosis factor (TNF) alpha production while increasing anti-inflammatory IL-1ra and IL-10. HSD prevented shock-induced norepinephrine surge with no effect on adrenal steroids. CONCLUSIONS: This first human trial evaluating the immunologic/anti-inflammatory effects of hypertonic resuscitation in trauma patients demonstrates that HSD promotes a more balanced inflammatory response to hemorrhagic shock, raising the possibility that similar to experimental models, HSD might also attenuate post trauma MOD. PMID- 16371738 TI - Lymphovascular invasion is associated with poor survival in gastric cancer: an application of gene-expression and tissue array techniques. AB - OBJECTIVES: To examine a population-based cohort for the association between clinicopathologic predictors of survival and immunohistochemical markers (IHC), and to assess changes in gene expression that are associated with lymphovascular invasion (LVI). SUMMARY BACKGROUND DATA: LVI has been associated with poor survival and aggressive tumor behavior. The molecular changes responsible for the behavior of gastric cancer have yet to be determined. Characterization of IHC markers and gene expression profiles may identify molecular alterations governing tumor behavior. METHODS: : Clinicopathologic and survival data of 114 patients were reviewed. Archival specimens were used to construct a multitumor tissue array that was subjected to IHC of selected protein targets. Correlation of IHC with tumor thickness (T status), LVI and prognosis was studied. Microarray analysis of fresh gastric cancer tissue was conducted to examine the gene expression profile with respect to LVI. RESULTS: In a multivariate analysis, nodal status (N), metastasis (M), and LVI were independent predictors of survival. LVI was associated with a 5-year survival of 13.9% versus 55.9% in patients in whom it was absent. LVI correlated with advancing T status (P = 0.001) and N status (P < 0.001). IHC staining of cyclooxygenase-2 (COX-2) correlated with T status, tumor grade, lymph node positivity, and IHC staining of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). Microarray analyses suggested differential expression of oligophrenin-1 (OPHN1) and ribophorin-II (RPNII) with respect to LVI. CONCLUSION: LVI was an independent predictor of survival in gastric cancer. Expression of COX-2 may facilitate tumor invasion through MMP-2 and MMP-9 activation. OPHN1 and RPN II appeared to be differentially expressed in gastric cancers exhibiting LVI. The reported function of OPHN1 and RPN II makes these gene products promising candidates for future studies involving LVI in gastric cancer. PMID- 16371739 TI - Predicting individual survival after gastric cancer resection: validation of a U.S.-derived nomogram at a single high-volume center in Europe. AB - OBJECTIVE: Validation of a U.S.-derived nomogram for individual prediction of disease-specific gastric cancer survival at a European institution. SUMMARY BACKGROUND DATA: One major issue of modern cancer treatment is the individualization of therapy. For gastric cancer, Kattan et al, at Memorial Sloan Kettering Cancer Center, New York, NY, developed a nomogram, allowing to predict individual patient risk of tumor-related death after R0 resection from basic patient-related variables. The validity of the nomogram has not yet been shown in patients from other institutions. The accuracy of the nomogram when applied to patients after having undergone R0 gastric cancer resection at a European high volume center was investigated. METHODS: Clinical data from patients who underwent R0 gastric cancer resection at Klinikum rechts der Isar, Technical University of Munich, Germany and fitted the respective derivation criteria were used for external validation (n = 862). Nomogram predictions for 60- and 108 month disease-specific survival were calculated for each patient and compared with actual survival. The concordance index was used as an accuracy measure. RESULTS: The bootstrap-corrected concordance index was 0.77 and was superior when compared with the predictive ability of International Union Against Cancer tumor stage (P < 0.008). Nomogram calibration was excellent for 60-month disease specific survival. Nomogram predictions showed the trend to underestimate survival in stage II/III disease of the MRI patients. CONCLUSIONS: The use of the nomogram created by Kattan et al is not only confined to the institution where it was created, but it can be adopted by other institutions with similar surgical strategies. PMID- 16371740 TI - Prevention of biliary lesions that may occur during radiofrequency ablation of the liver: study on the pig. AB - OBJECTIVE: To prevent bile duct injury by using a cold 5% glucose isotonic solution cooling in the bile ducts when radiofrequency (RF) is performed in a porcine model. SUMMARY BACKGROUND DATA: Complications that may arise during liver RF ablation include biliary stenosis and abscesses. METHODS: The RITA 1500 generator was used for the experiments. Two lesions were performed in the left liver. The pigs were killed 1 or 3 weeks after the procedure. An ex vivo cholangiogram was obtained by direct injection into the main bile duct. Samples of RF lesions, of liver parenchyma near and at a distance from the RF lesions, underwent pathologic studies. Two groups of 20 pigs each were treated: one without perfusion of the bile ducts and the other with perfusion of cold 5% glucose isotonic solution into the bile ducts. The Pringle maneuver was used in 50% of the RF procedures. Radiologic lesions were classified as biliary stenosis, complete interruption of the bile duct, or extravasation of the radiologic contrast liquid. RESULTS: Histologic lesions of the bile ducts were observed near the ablated RF lesion site and at a distance from the RF lesions when a Pringle maneuver was performed. Radiologic and histologic lesions of the bile ducts were significantly reduced (P < 0.0001) when the bile ducts were cooled. CONCLUSIONS: Cooling of the bile ducts with a cold 5% glucose isotonic solution significantly protects the intrahepatic bile ducts from damages caused by the heat generated by RF when performed close to the bile ducts. PMID- 16371741 TI - A clinical outcome-based prospective study on venous thromboembolism after cancer surgery: the @RISTOS project. AB - SUMMARY BACKGROUND DATA: The epidemiology of venous thromboembolism (VTE) after cancer surgery is based on clinical trials on VTE prophylaxis that used venography to screen deep vein thrombosis (DVT). However, the clinical relevance of asymptomatic venography-detected DVT is unclear, and the population of these clinical trials is not necessarily representative of the overall cancer surgery population. OBJECTIVE: The aim of this study was to evaluate the incidence of clinically overt VTE in a wide spectrum of consecutive patients undergoing surgery for cancer and to identify risk factors for VTE. METHODS: @RISTOS was a prospective observational study in patients undergoing general, urologic, or gynecologic surgery. Patients were assessed for clinically overt VTE occurring up to 30 +/- 5 days after surgery or more if the hospital stay was longer than 35 days. All outcome events were evaluated by an independent Adjudication Committee. RESULTS: A total of 2373 patients were included in the study: 1238 (52%) undergoing general, 685 (29%) urologic, and 450 (19%) gynecologic surgery. In hospital prophylaxis was given in 81.6% and postdischarge prophylaxis in 30.7% of the patients. Fifty patients (2.1%) were adjudicated as affected by clinically overt VTE (DVT, 0.42%; nonfatal pulmonary embolism, 0.88%; death 0.80%). The incidence of VTE was 2.83% in general surgery, 2.0% in gynecologic surgery, and 0.87% in urologic surgery. Forty percent of the events occurred later than 21 days from surgery. The overall death rate was 1.72%; in 46.3% of the cases, death was caused by VTE. In a multivariable analysis, 5 risk factors were identified: age above 60 years (2.63, 95% confidence interval, 1.21-5.71), previous VTE (5.98, 2.13-16.80), advanced cancer (2.68, 1.37-5.24), anesthesia lasting more than 2 hours (4.50, 1.06-19.04), and bed rest longer than 3 days (4.37, 2.45 7.78). CONCLUSIONS: VTE remains a common complication of cancer surgery, with a remarkable proportion of events occurring late after surgery. In patients undergoing cancer surgery, VTE is the most common cause of death at 30 days after surgery. PMID- 16371742 TI - Incidence and prevention of venous thromboembolism in patients undergoing breast cancer surgery and treated according to clinical pathways. AB - OBJECTIVE: To minimize treatment variations, we have implemented clinical pathways for all breast cancer patients undergoing surgery. We sought to determine the incidence of postoperative venous thromboembolism (VTE) in patients treated on these pathways. SUMMARY BACKGROUND DATA: Cancer patients have an increased risk of VTE because of a hypercoagulable state. The risk of VTE following breast cancer surgery is not well established. METHODS: We retrospectively reviewed prospectively collected data for all patients who underwent breast cancer surgery and were treated on the clinical pathways with mechanical antiembolism devices and early ambulation in the postoperative period between January 2000 and September 2003. RESULTS: During the study period, 3898 patients underwent 4416 surgical procedures. Seven patients with postoperative VTE within 60 days were identified, for a rate of 0.16% per procedure. Six patients presented with only a deep venous thrombosis or a pulmonary embolism; 1 patient had both. The median time from surgery to diagnosis of VTE was 14 days (range, 2-60 days; mean, 22 days). No relationship was identified between stage of breast cancer or type of breast surgery and development of VTE. Two (29%) of the 7 patients with VTE had received neoadjuvant chemotherapy. VTE treatment consisted of subcutaneous low-molecular-weight heparin (n = 5) or intravenous heparin (n = 2) followed by warfarin. There were no deaths. CONCLUSIONS: VTE following breast cancer surgery is rare in patients who are treated on clinical pathways with mechanical antiembolism devices and early ambulation in the postoperative period. We conclude that systemic VTE prophylaxis is not indicated in this group of patients. PMID- 16371743 TI - Detecting pheochromocytoma: defining the most sensitive test. AB - OBJECTIVE: To define the most sensitive biochemical test to establish the diagnosis of pheochromocytoma and also to assess the potential role of iodine 131 labeled metaiodobenzylguanidine scintigraphy (I-MIBG) in the diagnosis of this tumor. SUMMARY BACKGROUND DATA: Pheochromocytoma is a rare, catecholamine producing tumor with preferential localization in the adrenal gland. Despite its importance, the most sensitive test to establish the diagnosis remains to be defined. METHODS: Prospective data collection was done on patients with pheochromocytoma treated at the Duke University Medical Center and the Durham Veterans Affairs Medical Center, Durham, NC. All urinary, plasma, and platelet analyses were highly standardized and supervised by one investigator (J.M.F.). I MIBG scans were independently reviewed by 2 nuclear medicine physicians. RESULTS: A total of 152 patients (55.3% female) were enrolled in the present analysis. Patients were predominantly white (73.7%). Spells (defined as profuse sweating, tachycardia, and headache) and hypertension at diagnosis were present in 51.4% and 66.6%, respectively. Bilateral disease was found in 12.5%, malignant pheochromocytoma in 29.6%, and hereditary forms in 23.0%. The most sensitive tests were total urinary normetanephrine (96.9%), platelet norepinephrine (93.8%), and I-MIBG scintigraphy (83.7%). In combination with I-MIBG scintigraphy, platelet norepinephrine had a sensitivity of 100%, plasma norepinephrine/MIBG of 97.1%, total urine normetanephrine/MIBG of 96.6%, and urine norepinephrine/MIBG of 95.3%. CONCLUSIONS: The tests of choice to establish the diagnosis of pheochromocytoma are urinary normetanephrine and platelet norepinephrine. A combination of I-MIBG scintigraphy and diagnostic tests in urine, blood, or platelets does further improve the sensitivity. We thus advocate performing an MIBG scan if the diagnosis of pheochromocytoma is clinically suspected and catecholamine measurements are within the normal range. PMID- 16371744 TI - Gut hormone profiles following bariatric surgery favor an anorectic state, facilitate weight loss, and improve metabolic parameters. AB - OBJECTIVE: To study the effect of bariatric surgery on the entero-hypothalamic endocrine axis of humans and rodents. BACKGROUND: Bariatric surgery is the most effective obesity treatment as it achieves substantial and sustained weight loss. Glycemic control and enhanced satiation improve before substantial weight loss occurs. Gut peptides, acting both peripherally and centrally, contribute to glycemic control and regulate food intake. METHODS: We examined meal-stimulated responses of insulin, ghrelin, peptide YY (PYY), glucagon-like-peptide-1 (GLP-1), and pancreatic polypeptide (PP) in humans and rodents following different bariatric surgical techniques. RESULTS: Compared with lean and obese controls, patients following Roux-en-Y gastric bypass (RYGB) had increased postprandial plasma PYY and GLP-1 favoring enhanced satiety. Furthermore, RYGB patients had early and exaggerated insulin responses, potentially mediating improved glycemic control. None of these effects were observed in patients losing equivalent weight through gastric banding. Leptin, ghrelin, and PP were similar in both the surgical groups. Using a rodent model of jejuno-intestinal bypass (JIB), we showed elevated PYY and GLP-1 in JIB rats compared with sham-operated rats. Moreover, exogenous PYY reduced food intake and blockade of endogenous PYY increased food intake. Thus, higher plasma PYY following JIB may contribute to reduced food intake and contribute to weight loss. CONCLUSIONS: Following RYGB and JIB, a pleiotropic endocrine response may contribute to the improved glycemic control, appetite reduction, and long-term changes in body weight. PMID- 16371745 TI - The use of beta-adrenergic blockade in preventing trauma-induced hepatomegaly. AB - OBJECTIVE: The objective of this study was to test the hypothesis that hepatomegaly in burned children can be attenuated or reversed by blocking lipolysis and reducing free fatty acids delivered to the liver. SUMMARY BACKGROUND DATA: Accelerated lipolysis in severely burned children has been shown to play an important role in the accumulation of hepatic TGs. Severely burned children who survive 10 days or more after injury commonly have enlarged livers often twice or more normal size for their sex, age, and weight. METHODS: Ninety eight children, 2 to 18 years of age, with burns covering more than 40% of their body surface and who received either propranolol (beta-adrenergic blockade) or placebo were studied. Liver weights were measured by ultrasonic scanning. Body composition changes were identified by dual-image x-ray absorptiometry and validated by whole-body potassium-40 scintillation counting. Discarded abdominal cutaneous adipose tissue was collected before and after propranolol or placebo for microarray analysis. RESULTS: In 80% of severely burned children studied not receiving propranolol, liver sizes increased by 100% or more while 86% of burned children receiving propranolol showed a decrease or no change in liver size over the same period of time after injury. Gene expression patterns of adipose tissue after propranolol treatment showed that all of the identified genes related to lipid metabolism were down-regulated. CONCLUSIONS: Data reported here support the hypothesis that beta-adrenergic blockade can reduce delivery of fatty acids to the liver and hepatic congestion commonly found in severely burned children by inhibiting lipolysis and reducing hepatic blood flow. PMID- 16371746 TI - Delta V as a predictor of significant injury for children involved in frontal motor vehicle crashes. AB - OBJECTIVE: The objective of this study was to examine the association between delta V and risk of injury to children involved in frontal motor vehicle crashes. BACKGROUND: Previous studies, primarily focused on adult occupants, have demonstrated the relationship between crash severity, as measured by delta V and injury severity. As children have unique safety needs, these results cannot be directly applied to the pediatric population. METHODS: Case series crash investigation data and clinical injury information were reviewed from a child focused crash surveillance system. Analyses were performed examining the relationship between the estimated delta V and any AIS > or = 2 or any AIS > or = 3 injury. RESULTS: Detailed crash investigation and clinical data were available on 407 children involved in 235 frontal crashes. The average delta V for all crashes was 29 +/- 16.9 kph [18 +/- 10.5 mph (range, 5-123 kph)]. Delta V was strongly and positively associated with the odds of both an AIS > or = 2 and AIS > or = 3 injury (P < 0.0001). The adjusted odds of at least one AIS > or = 2 injury increased on average by 56% (95% confidence interval [CI], 33%-85%) for each 10 kph increase in delta V. Similarly, the adjusted odds of at least one AIS > or = 3 injury increased on average by 67% (95% CI, 40%-102%) for each 10 kph increase in delta V. The delta V at which 50% of child occupants would be expected to sustain any AIS > or = 2 injury was 37 kph [23.0 mph (95% CI, 32-45 kph)], and any AIS > or = 3 injury was 63 kph [39.1 mph (95% CI, 51-infinity kph)]. CONCLUSION: Delta V is strongly predictive of injury risk for child occupants. As many newer generation cars are now fitted with event data recorders, this information is increasingly available and may become useful as a clinical predictor. PMID- 16371747 TI - Fatal and nonfatal hemorrhagic complications of living kidney donation. AB - OBJECTIVE: After anecdotal reports of severe hemorrhage from failure of surgical clips to sustain closure of renal artery stumps in live donor nephrectomies were received, this study was designed to identify specific surgical techniques that are associated with an increased risk of failure to control bleeding and might represent opportunities to improve patient safety. BACKGROUND: Preventing complications for living kidney donors must be paramount in addressing end-stage renal failure through living kidney donation. Major hemorrhage from technical failure, albeit an infrequent occurrence, can cause significant, yet preventable, morbidity or death. Open and laparoscopic approaches to living kidney donation use several vascular control methods, some of which may be more prone to failure and life-endangering hemorrhage than others. METHODS: To define hemorrhagic complications of living kidney donation, a survey was sent to all 893 surgeon members of the American Society of Transplant Surgeons. Descriptive and bivariate analyses were used to ascertain study participant characteristics, most frequently used vascular control techniques, and incidence of events (death, transfusion, reexploration or conversion to open nephrectomy, or contralateral [remaining kidney] renal failure). Outcomes of hemorrhage and comments by respondents were sought as were data from other sources. RESULTS: In 213 surveys returned (24%), 66 and 39 episodes of arterial and venous hemorrhage were reported, respectively. Among arterial control problems, 2 resulted in donor death and 2 resulted in renal failure; 19 episodes required transfusion. Open conversions in laparoscopic nephrectomy or late reoperations for hemorrhage were reported for 29 cases. Locking and standard clips applied to the renal artery were associated with the greatest risks. CONCLUSIONS: Significant hemorrhagic complications occur with living kidney donation in both open and laparoscopic approaches. Loss of arterial control jeopardizes donor life and health, especially when it occurs in the postoperative period. Vascular transfixion provides the best vascular control of major vessels. PMID- 16371748 TI - Awake nonresectional lung volume reduction surgery. AB - OBJECTIVE: To assess the feasibility, safety, and early results of awake lung volume reduction surgery (LVRS) performed under thoracic epidural anesthesia by a new nonresectional technique. SUMMARY BACKGROUND DATA: So far, resectional LVRS under general anesthesia and one-lung ventilation is the more frequently used technique, but procedure-related morbidity has been considerable. METHODS: The study cohort included 12 patients undergoing unilateral awake LVRS. Evaluated parameters included technical feasibility and anesthesia satisfaction scored into 4 grades (from 1 = poor to 4 = excellent), global operating room time, and arterial carbon dioxide tension (PaCO2). In addition, 6-month changes in outcome measures, including forced expiratory volume in 1 second (FEV1), residual volume (RV), 6-minute walking test (SMWT), and dyspnea index were recorded. Perioperative and 6-month results were comparable with those of a control group undergoing unilateral resectional LVRS. RESULTS: Technical feasibility was excellent to satisfactory in 11 patients. One patient required conversion to one lung ventilation. Differences between the awake and control group included global operating room time (90 +/- 17 minutes versus 145 +/- 19 minutes, P < 0.00001); PaCO2 24 hours after surgery (45 +/- 6 mm Hg versus 49 +/- 6 mm Hg, P = 0.02); and hospital stay (7.8 +/- 5 days versus 11.7 +/- 4 days, P = 0.02). Significant (P < 0.002) improvements occurred at 6 months in FEV1 (0.31 +/- 0.17 L), RV ( 1.41 +/- 0.7 L), SMWT (73 +/- 25 m), and dyspnea index (-1.3 +/- 0.5) and were comparable with those of the control group. CONCLUSIONS: In this study, awake nonresectional LVRS proved feasible and safe. This new modality was associated with a faster recovery and satisfactory 6-month outcome, which did not differ from that of resectional LVRS. PMID- 16371749 TI - Complete versus selective portal triad clamping for minor liver resections. PMID- 16371750 TI - Delayed massive hemorrhage after pancreatic and biliary surgery: embolization or surgery? PMID- 16371752 TI - Transjugular intrahepatic portosystemic shunt versus H-graft portacaval shunt in the management of bleeding varices: a cost-benefit analysis. PMID- 16371754 TI - Drainage and other risk factors for leakage after anterior resection in rectal cancer patients: a prospective study of 978 patients. PMID- 16371756 TI - Pelvic drainage should be a routine for TME with or without radiation. PMID- 16371759 TI - Differences by age and race/ethnicity in knowledge about hypercholesterolemia. AB - This investigation sought to identify gaps in patients' knowledge about hypercholesterolemia and lipid-lowering therapy in an inner-city US population. Chart reviews and interviews were conducted for drug-treated hypercholesterolemic patients at 3 hospital-based cardiology practices in Bronx, NY. Univariate and multivariate analyses were used to assess whether race/ethnicity, sex, and age were predictors of knowledge. Subjects (n = 467, age 65.3 years) were 55% female, 38% Hispanic, 32% black, and 25% white. Most recognized hypercholesterolemia as a cause of heart disease (88%) and stroke (76%), although older subjects (>70 years) and Hispanic subjects, particularly non-English speakers, were significantly less likely to know about cardiovascular disease risks associated with hypercholesterolemia. Only 14% recalled their recent cholesterol levels, and recall was significantly lower among black, Hispanic, and older subjects. Overall, 31% expected to require lipid-lowering medications indefinitely in the future, while 66% did not know how long they would need to continue using medications. Blacks and Hispanics were significantly less likely to believe that they would need to continue taking medications indefinitely. Achieved lipid levels and self-reported medication adherence were relatively favorable although were unrelated to knowledge levels. In summary, among hypercholesterolemic patients in an urban population, relatively few knew their own cholesterol levels or expected to require medications indefinitely in the future. Older patients, black and Hispanic patients, and non-English speakers were significantly less likely to have accurate knowledge about hypercholesterolemia. Therefore, patient and community education efforts targeted to specific populations may lead to improved management of hypercholesterolemia in inner-city regions. PMID- 16371760 TI - The cost-effectiveness of argatroban treatment in heparin-induced thrombocytopenia: the effect of early versus delayed treatment. AB - A decision-tree analysis was used to estimate the average cost per patient using the direct thrombin inhibitor argatroban for early treatment (<48 hours after thrombocytopenia onset) compared with delayed treatment (> or =48 hours after thrombocytopenia onset) of immune-mediated heparin-induced thrombocytopenia (HIT) with or without thrombosis. Clinical probability data used to populate the model were obtained from argatroban clinical trials and from published clinical literature. Resource utilization data and cost data were also obtained from available literature, the 2003 Physician's Fee Reference, the Healthcare Cost and Utilization Project 2000, the 2003 Drug Topics RedBook, and a modified Delphi panel. The total per-patient cost included hospital days, diagnostic tests, heparin, argatroban, major hemorrhagic events, and patient outcomes (ie, amputation, new thrombosis, stroke, or death), multiplied by the probability of each event. The incremental cost-effectiveness ratio was calculated by dividing the incremental cost between patients with and without argatroban treatment by the incremental effectiveness, or the cost per new thrombosis event avoided. The mean cost per HIT patient without thrombosis who did not receive argatroban was $38,046. The mean cost decreased by 6.85% for patients who were treated earlier with argatroban therapy (average cost, $35,441), representing a $2605 saving per patient compared with those not treated with argatroban. For those receiving delayed argatroban therapy, the mean cost increased by $9024 per patient compared with those receiving early treatment with argatroban. The mean cost for HIT patients with thrombosis who did not receive argatroban was $48,101, which was 9.0% higher than for those receiving early argatroban therapy, representing a $3957 savings per patient. For HIT with thrombosis, mean costs increased by 18.2% in patients whose argatroban was delayed, representing a cost increase of $8020 per patient compared with early treatment (mean cost $44,144 for early treatment and $52,164 for delayed treatment). The results of this analysis support the recommendation to initiate early argatroban treatment upon suspicion of HIT to reduce the thrombotic consequences of HIT and associated healthcare costs. Argatroban therapy should not be delayed pending the results of HIT diagnostic tests. PMID- 16371761 TI - Increased prevalence of coronary artery disease, silent myocardial ischemia, complex ventricular arrhythmias, atrial fibrillation, left ventricular hypertrophy, mitral annular calcium, and aortic valve calcium in patients with chronic renal insufficiency. AB - Cardiovascular morbidity and mortality is high in patients with chronic renal insufficiency. Patients with chronic renal insufficiency have an increased prevalence of coronary artery disease, silent myocardial ischemia, complex ventricular arrhythmias, atrial fibrillation, left ventricular hypertrophy, mitral annular calcium, and aortic valve calcium than patients with normal renal function. These risk factors for cardiovascular morbidity and mortality contribute to the increased incidence of cardiovascular morbidity and mortality seen in patients with chronic renal insufficiency. PMID- 16371762 TI - Aspirin resistance: mechanisms and clinical implications. AB - Acetylsalicylic acid (aspirin) has been shown to irreversibly interfere with platelet function, an effect that is associated with a reduction in morbid and mortal arterial thrombotic events in multiple clinical studies. This clinical benefit appears to be attenuated by resistance to the antiplatelet effects of aspirin in up to 35% of patients. The mechanisms for aspirin resistance are multifactorial and include noncompliance with aspirin therapy, diabetes mellitus, cell-cell and drug-drug interactions, genetic polymorphisms, and coronary artery disease. It has not been determined what the best laboratory procedure is to screen for aspirin resistance. Those individuals at high risk for aspirin resistance might best be treated with an additional oral antiplatelet drug (eg, clopidogrel) to achieve maximal protection against arterial thrombotic events. PMID- 16371763 TI - Angiotensin II receptor blockers in congestive heart failure. AB - The benefits of angiotensin-converting enzyme (ACE) inhibitors for the treatment of congestive heart failure (CHF) are well-established. A newer class of medications, angiotensin II receptor blockers (ARBs), may be a suitable replacement for ACE inhibitors as a result of a more complete inhibition of angiotensin II and better tolerability among patients. To examine the current literature on the efficacy and safety of ARBs in the setting of CHF, a Medline search was conducted of the English language literature for the years 1987 to 2005. Clinical trials that reported data on cardiac outcomes were reviewed. The earlier trials were direct ARB to ACE inhibitor comparisons (ELITE I and ELITE II). These studies indicated that ARBs do not confer an improvement in cardiac outcomes over ACE inhibitors. RESOLVD, Val-HeFT, and the 3 separate trials of the CHARM program investigated the addition of an ARB to standard therapy. The RESOLVD trial showed no significant differences in clinical events among ACE inhibitor, ARB, and their combination. Although no mortality benefit was evident in the Val-HeFT trial, a substantial reduction in CHF rehospitalizations was reported among patients who were not receiving ACE inhibitor therapy. The CHARM Overall program demonstrated a significant benefit in cardiovascular death and hospital admissions for CHF with the addition of ARB to standard therapy, a benefit that was more pronounced in patients with depressed left ventricular ejection fraction. In the setting of CHF, rates of cardiac outcomes do not differ substantially between ARBs and ACE inhibitors. However, their combination may improve outcomes for patients with CHF. PMID- 16371764 TI - Peripartum cardiomyopathy. AB - Peripartum cardiomyopathy (PPCM) is a rare cardiac disorder associated with high rates of mortality that occurs during the peripartum period. PPCM is recognized as a distinct entity, separate from preexisting cardiomyopathies that are worsened by the stressors of pregnancy. To date, its etiology is unknown, although several theories are under investigation in an effort to provide more information regarding available treatment options. A multidisciplinary review of PPCM held by the National Heart, Lung, and Blood Institute, in conjunction with the Office of Rare Disease of the National Institutes of Health, in April 1997 reviewed the current knowledge and developed recommendations for areas of further research and education about PPCM. Since then, there have been some promising research testing hypotheses regarding the etiology of PPCM and advancements in possible treatment options. However, despite these efforts, knowledge and treatment recommendations about PPCM are still generally unchanged, whereas mortality rates remain high. This article attempts to provide an updated, comprehensive review about PPCM and draw attention to areas in need of further research. PMID- 16371765 TI - MRI case: seventeen-year-old with systemic hypertension. PMID- 16371766 TI - Prosthetic valve endocarditis caused by Acinetobacter lwoffi: a case report and review. AB - We are reporting a case of recurrent prosthetic-valve endocarditis (PVE) caused by an unusual pathogen. The patient suffered 2 consecutive relapses of Acinetobacter lwoffi bacteremia, although he had completed a full course of treatment with antibiotics to which the microorganism was susceptible. He was finally successfully operated with replacement of the infected aortic valve. Acinetobacter spp are relatively low-grade but potentially virulent pathogens, and endocarditis caused by these species can be fulminant, accompanied by septic complications, and fatal. Although some patients with relapsed PVE may respond to a second course of antibiotics and medical treatment rather than early valve replacement is suggested in A lwoffi PVE, combined antibiotic treatment and early surgical intervention may be considered as the first option in these patients. There are only a few cases of Acinetobacter endocarditis in the literature, and it is the first case reported in Greece to our knowledge. PMID- 16371767 TI - Acute coronary syndrome after infliximab infusion. AB - The TNFalpha inhibitor infliximab is widely used in the treatment of rheumatoid arthritis and Crohn disease. Mild infusion reactions consisting of low-grade fever, headache, nausea, and fatigue are common, but we describe for the first time the occurrence of an acute coronary syndrome during infliximab administration. This case alerts infusion centers to consider the possibility that chest pain and dyspnea during infliximab infusion can represent a myocardial infarction, even in younger patients without a history of cardiac disease. PMID- 16371769 TI - Wound dehiscence after penetrating keratoplasty. AB - PURPOSE: To assess the patient characteristics, risk factors, and outcomes of penetrating keratoplasty wound dehiscence. METHODS: Retrospective chart review of 30 eyes of 29 patients with corneal grafts who underwent repair of penetrating keratoplasty wound dehiscence from January 1, 1998 to December 31, 2003, followed on the Cornea Service at Wills Eye Hospital. RESULTS: The mean time from penetrating keratoplasty to wound dehiscence was 7.5 years (range 1 week to 31 years). The mean age at time of wound dehiscence was 66 years (range 28-98 years). One eye developed wound dehiscence following suture removal, whereas the remaining 29 eyes sustained trauma-induced dehiscences. Falls were the most common mechanism of trauma, especially in the elderly population. There was a wide range of visual outcomes in the 21 patients followed for 1 year, with 4 patients maintaining best corrected visual acuity between 20/20 and 20/40 and 5 patients with light perception vision at 1 year. There were no cases of endophthalmitis. CONCLUSIONS: Patients with corneal transplants have a life-long risk for wound dehiscence. This complication may be reduced by the regular use of eye protection in all corneal transplantation patients. PMID- 16371768 TI - In vivo investigations of the corneal epithelium with the confocal Rostock Laser Scanning Microscope (RLSM). AB - BACKGROUND: The confocal tandem scanning microscope was first used in 1985 by Lemp et al for in vitro and in 1990 by Cavanagh et al for in vivo investigation of human eyes. The aim of this study was to investigate the cells of the central and the peripheral portions of the corneal epithelium and to measure corneal epithelium thickness and the total thickness of the corneas of our volunteers with the new Rostock Laser Scanning Microscope. MATERIAL AND METHODS: A Heidelberg Retina Tomograph (HRT II) was used in combination with a water contact microscope lens (Zeiss, x63, 0.95), the Rostock cornea module (RCM) developed at our institute for the in vivo examination of the cornea. In this study, 92 eyes of 68 subjects between the ages of 15 and 88 years were examined. RESULTS: At the superficial cell layer, the average cell density in the central cornea was 840 +/ 295 cells/mm2, and in the periphery it was 833 +/- 223 cells/mm2. At the wing cell layer, the average cell density rises to 5070 +/- 1150 cells/mm2 in the central and to 5582 +/- 829 cells/mm2 in the peripheral cornea. At the basal cell layer, the cell density rises further to 8996 +/- 1532 cells/mm2 in the central and 10,139 +/- 1,479 cells/mm2 in the peripheral corneal epithelium. The average corneal thickness in the central region was found to be 545 +/- 25 microm, and 652 +/- 75 microm in the periphery. The average epithelium thickness was determined centrally to be 54 +/- 7 microm, and peripherally 61 +/- 5 micrim. CONCLUSIONS: The Rostock Scanning Laser Microscope offers a standardized, reproducible, safe, and fast diagnostic procedure for the evaluation of the corneal epithelium. This technology allows better image quality compared with confocal-slit scanning microscopes and produces a precise depth measurement. PMID- 16371770 TI - Surgical monovision and monovision reversal in LASIK. AB - PURPOSE: This study was designed to assess the success of surgical monovision in presbyopic patients. METHODS: A university refractive surgery center retrospective chart review of 82 patients who elected to undergo surgical monovision with laser in situ keratomileusis (LASIK) between January 2000 and January 2003 was conducted. Specific factors included for analysis included preoperative and postoperative defocus spherical equivalent, whether the patient underwent enhancements, whether the patient underwent a preoperative monovision trial with contact lens, and whether the patient underwent monovision reversal. RESULTS: Eighty-two patients who underwent LASIK for monovision were analyzed. Mean preoperative spherical equivalent in the distance-corrected eye was -4.07 (standard deviation (SD), 2.49); for the eye corrected for near vision, mean preoperative spherical equivalent was -4.10 (SD, 2.56). Postoperative spherical equivalent in the distance eyes was -0.01 (SD, 0.38) and in the near eyes -1.24 (SD, 0.91). There were 6 enhancements in the near eyes (7%) and 17 enhancements in the distance vision eyes (21%). This difference was statistically significant (P = 0.007). Thirty patients underwent a contact lens trial of monovision before LASIK, and none of those patients elected monovision reversal. There were 52 patients who did not undergo a contact lens monovision trial before LASIK monovision, and 2 of these patients underwent monovision reversal. Monovision success in this population was 97.6%. CONCLUSION: Surgical monovision can help presbyopic patients achieve their goal of reduced dependence on spectacles. A trial of monovision contact lenses or spectacles may be important in helping surgeons select patients for successful surgical monovision. PMID- 16371771 TI - Ocular surface reconstruction for thermal burns caused by fireworks. AB - PURPOSE: To describe 6 eyes with severe firework-related ocular surface injuries that were treated by limbal stem cell transplantation combined with amniotic membrane transplantation (AMT). METHODS: Six eyes of 6 patients with firework related ocular injuries were retrospectively studied. All subjects were men, with age ranging from 9 to 26 years. All patients were in chronic stage of thermal injury, and all had total limbal stem cell dysfunction and subsequent conjunctival fibrosis. They had limbal stem cell transplantation (1 had an allograft and 5 had autograft transplantation) combined with AMT to restore vision. Mitomycin C was used in 1 eye intraoperatively. RESULTS: After ocular surface reconstruction using limbal stem cell transplantation combined with AMT, corneal epithelialization was achieved in 4 eyes after a mean duration of 20.8 +/ 12.2 days (range, 4-33 days). After a mean follow-up period of 36 months (range, 5-87 months), marked reduction of fibrosis was noted in all eyes. At last examination, the corneal surface was covered by corneal-phenotype epithelium in 4 eyes, and the remaining 2 eyes had conjunctivalization. The cornea was optically clear in 3 eyes, with corrected visual acuity of 20/100 in these 3 eyes. Both eyes that developed conjunctivalization had massive fibrosis preoperatively and severe eyelid deformities. CONCLUSION: Although severe firework-related ocular surface injuries were difficult to treat, limbal stem cell transplantation combined with AMT enable some success in cosmetic and visual outcome. Management of eyelid abnormalities and ocular surface fibrosis seemed to be a key to success in visual rehabilitation. PMID- 16371772 TI - Late changes in refraction, pachymetry, visual acuity, and corneal topography after penetrating keratoplasty. AB - PURPOSE: To analyze the late changes in refraction, corneal topography, and pachymetry after penetrating keratoplasty. METHODS: We have retrospectively analyzed data of 64 eyes of 56 patients with a clear corneal graft and no other ocular diseases. The eyes were examined soon and several years after all sutures were removed, at 19.5 +/- 3.1 months and 76.8 +/- 25.2 months, respectively, after keratoplasty. All eyes were examined using the EyeSys 2000 videokeratograph and the Tomey SP2000 ultrasonic pachymeter. Corneal surface regularity was studied using the indices provided by the Holladay Diagnostic Summary (ie, asphericity coefficient Q; corneal uniformity index CU; predicted corneal acuity PCA). Irregular astigmatism was quantified using semimeridian data from videokeratographs (refractive power symmetry index). RESULTS: Visual acuity increased by an average of 0.35 +/- 0.93 lines (P = 0.002) between the 2 examinations, whereas no significant changes in refraction and videokeratoscopy (ie, power, indices, and irregular astigmatism) were observed (P > or = 0.08). Central corneal thickness significantly increased from 542 +/- 31 microm to 572 +/- 38 microm (P < 0.001). Change in BSCVA did not significantly correlate with the change in refraction, in topographic indices, or in irregular astigmatism (rs < or = 0.13; P > or = 0.16). CONCLUSION: Late after penetrating keratoplasty, best corrected visual acuity shows a small but statistically significant improvement, whereas refraction and corneal surface regularity are stable. This late improvement in visual acuity is not explained by current techniques. PMID- 16371773 TI - Microbial keratitis at extremes of age. AB - PURPOSE: To study the risk factors, clinical presentation, response to treatment and outcome of microbial keratitis occurring in patients at extremes of age (ie, children and the elderly) and to compare them with microbial keratitis affecting the general adult population. METHODS: In this prospective, nonrandomized, analytical clinical study, all patients presenting with microbial keratitis and undergoing microbiological workup over a 1-year period were evaluated using a dedicated corneal ulcer protocol. Patients were divided into 3 groups based on age: (1) pediatric group if the age was 16 years or younger, (2) elderly group if the age was 65 years or older, and (3) control group if the age was between 17 and 64 years. Features studied included microbiological profile, predisposing factors, clinical presentation, response to treatment, and final visual outcome. RESULTS: A total of 269 eyes of 269 patients were studied (26 in the pediatric group, 55 in the elderly group, and 188 in the control group). Culture positivity rates were similar in all the groups. The elderly and control groups had a similar incidence of fungal and bacterial keratitis, but the pediatric group had a significantly lower incidence of fungal keratitis (P = 0.001). Trauma was the most common predisposing factor in all groups. Elderly patients tended to present with a higher incidence of central ulcers (P = 0.04), severe ulcers (P = 0.04), and poor visual acuity (P = 0.003) as compared with the control group. The percentage of ulcers healing with medical treatment alone was significantly more in the pediatric group (P = 0.004), while the incidence of poor visual outcome was greater in the elderly group (P = 0.006) as compared with the control group. CONCLUSIONS: Microbial keratitis in pediatric patients is more likely to be of bacterial etiology, and nonsevere ulcers in this age group have a better chance of resolution with medical therapy alone when compared with that in the general adult population. Conversely, elderly patients with microbial keratitis tend to present with severe, central ulcers with a significant risk of having a poor visual outcome. PMID- 16371774 TI - Subpalpebral lavage antibiotic treatment for severe infectious scleritis and keratitis. AB - OBJECTIVE: To report the subpalpebral lavage therapy for the treatment of infectious scleritis and keratitis. METHOD: Six patients were admitted for treatment of severe infectious scleritis and keratitis and were initiated on the subpalpebral lavage system after showing no improvement with topical fortified antibiotics. A continuous antibiotic lavage was applied until clinical sterility was achieved and topical steroids were gradually added to control concomitant inflammation. RESULTS: All 6 patients had resolution of their infections and achieved a stable ocular surface with no inflammation. One patient required a corneal transplant for active inflammation and corneal thinning, 1 had a transplant for a dense central corneal scar, and 1 patient underwent corneal transplant and cataract extraction. One case was a Pseudomonas keratitis in a blind eye, which rapidly resolved and has remained stable. Four patients required additional surgeries, which included 3 corneal transplants, 2 cataract extractions, and 1 glaucoma aqueous shunt. CONCLUSION: Continuous irrigation of the eye can improve scleral penetration of antibiotics. Subpalpebral lavage provides continuous irrigation and may be effective in the treatment of infectious scleritis of a variety of etiologies. PMID- 16371775 TI - Clinical profile and early surgical complications in the Cornea Donor Study. AB - PURPOSE: The Cornea Donor Study was designed to investigate the safety and efficacy of older donor corneal tissue compared with younger donor tissue in recipient eyes at moderate risk to the graft from progressive endothelial failure. Baseline patient data, including indications for transplant, intraoperative complication rates, and early postoperative complication rates are described herein. METHODS: This study was a multicenter prospective, double masked, controlled clinical trial. RESULTS: Fuchs dystrophy was the most common indication for corneal transplantation (61%). Intraoperative complications occurred in 33 (3%) patients. A persistent epithelial defect was the most commonly reported postoperative complication, occurring in 92 patients (8%). CONCLUSION: Intraoperative and postoperative complication rates were low. There was no apparent association between donor or recipient age and either intraoperative or early postoperative complication rates. PMID- 16371776 TI - Efficacy of commercially available topical cyclosporine A 0.05% in the treatment of meibomian gland dysfunction. AB - OBJECTIVE: To investigate the efficacy of topical cyclosporine A 0.05% (tCsA) (Restasis, Allergan Pharmaceuticals) in the treatment of meibomian gland dysfunction (posterior blepharitis). METHODS: Thirty-three patients with symptomatic meibomian gland dysfunction were randomized in a prospective study to either tCsA or placebo (Refresh Plus preservative-free artificial tears), 2 times daily for 3 months. They were evaluated at baseline and at 1, 2, and 3 months for subjective symptoms and objective signs including meibomian gland inclusions, lid margin vascular injection, tarsal telangiectasis, fluorescein staining, tear breakup time, and Schirmer scores. RESULTS: Twenty-six patients completed the study. All patients were tested for ocular symptoms, lid margin vascularity, tarsal telangiectasis, meibomian gland inclusions, tear breakup time, and fluorescein staining. At the 3-month visit, the tCsA group showed a greater improvement in ocular symptoms than the placebo group, but this difference was not statistically significant. At the 3-month visit, several objective examination findings were statistically significantly (P < 0.05) improved in the tCsA group compared with the placebo group. These differences included lid margin vascular injection, tarsal telangiectasis, and fluorescein staining. The most significant finding (P = 0.001) was the greater decrease in the number of meibomian gland inclusions in the tCsA group compared with the placebo group. CONCLUSIONS: Topical CsA may be helpful in the treatment of meibomian gland dysfunction (posterior blepharitis). Topical CsA did not induce an improvement in the symptoms, but it did decrease the number of meibomian gland inclusions in patients with meibomian gland dysfunction. PMID- 16371777 TI - Mucins and contact lens wear. AB - PURPOSE: This study was designed to determine whether long-term tolerant contact lens (CL) wear causes changes in the expression of mucin mRNA by the conjunctival epithelium and mucin protein content in tears and to determine whether specific mucins adhere to contact lenses. METHODS: Twenty long-term (> or = 5 years ) and tolerant CL wearers (2 with hard and 18 with soft contact lenses) were compared with 23 non-CL wearers. One hour after CL removal, tear fluid was collected after instillation of 60 microL of sterile water onto the ocular surface, and protein concentration was determined. Impression cytology was performed on the bulbar temporal region of conjunctiva to collect cells for RNA isolation. Real-time polymerase chain reaction was performed using TaqMan primer and probes for MUC1, 4, 5AC, and 16. ELISA was performed on the collected tears to detect MUC5AC and the mucin carbohydrate epitope H185. For the analysis of adherent mucins on CL, discarded daily-wear contact lenses were collected, rinsed, and incubated overnight at 4 degrees C in mucin isolation buffer. Immunoblot analysis of adherent mucins was performed to detect MUC1, 4, 5AC, 16, and H185. RESULTS: No significant changes in the levels of mucin mRNA from impression cytology samples were detected when comparing CL and non-CL wearers. The amount of total protein in tears collected from CL wearers (39.9 +/- 27.2 microg) was significantly less than that from non-CL wearers (95.1 +/- 73.8 microg, P = 0.001). The level of MUC5AC mucin and the H185 epitope in tears per unit protein in CL wearers was not significantly different from non-CL wearers. Low levels of membrane-associated mucins, the secreted mucin MUC5AC, and the carbohydrate epitope, H185, were detected in protein extracts from discarded CLs. Compared with MUC1, 4, and 5AC, there was less MUC16 adherent to the CLs. CONCLUSION: Neither mucin mRNA expression by conjunctival epithelia nor mucin content per unit protein in tears was altered by long-term tolerant CL wear; however, the amount of protein in the tears was significantly less. Shed membrane-associated mucins and the goblet cell mucins adhere to CLs. PMID- 16371778 TI - Continuous measurement of corneal dehydration with online optical coherence pachymetry. AB - PURPOSE: Online optical coherence pachymetry (online OCP) allows continuous central corneal thickness measurements over time. In this study, the effect of dehydration on corneal tissue was investigated with online OCP. METHODS: Twelve eyes of 11 patients were examined with online OCP, and the central corneal thickness was registered over 5 minutes after insertion of an eyelid speculum. RESULTS: Online OCP measurements revealed no decrease in reproducibility after 5 minutes of dehydration. The initial mean central corneal thickness was 538 +/- 48 microm. After 5 minutes the central corneal thickness decreased to 483 +/- 43 microm (P = 0.001). This corresponded to a mean corneal thinning of 55 +/- 4 microm (10.2%) at a rate of 0.19 microm/s. CONCLUSIONS: Online OCP was suitable for continuous measurements of corneal changes caused by dehydration. These dehydration effects should be particularly considered in refractive corneal surgery. PMID- 16371779 TI - Reducing contamination when removing and storing corneas: a multidisciplinary, transversal, and environmental approach. AB - PURPOSE: The combination of a shortage of cornea grafts in France and a national average contamination rate of 9% to 10%, has led us to search for the origins of this contamination. The objective of our study was to reduce the number of unusable grafts resulting from contamination of corneas in organ culture. METHODS: An external audit was carried out by an independent pharmacist on the removal conditions and treatment procedures for corneas. An environmental study was carried out, consisting of microbiological sampling of the corneas of donors who just died (<24 hours) as well as water and air samples in the premises used for removal. The Cornea Bank's procedures were submitted to a microbiological risk analysis using the "failure mode effects and criticity analysis" (FMECA) method. RESULTS: The critical contamination periods were found to be before removal, during mortuary washing and during decontamination of the conjunctival cul-de-sac at the removal stage. The corrective measures taken have reduced contamination rates by half in 1 year. CONCLUSION: Highlighting the sources of contamination has led to the implementation of effective targeted and low-cost measures that have allowed us to reduce significantly the number of cornea graft losses as a result of bacterial and fungal contamination. PMID- 16371780 TI - Progressive replacement of oral mucosa by conjunctiva in osteo-odonto keratoprosthesis: preliminary observations. AB - PURPOSE: In a Strampelli osteo-odonto-keratoprosthesis, a patch of oral mucosa is frequently used to cover the ocular surface after implantation of an osteodental lamina into the eye. In many cases, gross modifications in the eye covering become apparent a few years later. The aim of this study was to investigate the histologic findings in the clinically modified ocular surface. METHOD: Biopsies were performed in 7 patients at the junction between the osteodental acrylic lamina and surrounding modified oral mucosa, during surgery for local plastic reconstruction or positioning of antiglaucoma silicone tubes. Specimens were examined by light microscopy. RESULTS: Six of the 7 clinically modified specimens corresponded microscopically to conjunctiva. Typical oral mucosa could still be observed overlying the osteodental acrylic lamina. CONCLUSIONS: The production of local regulatory factors is a possible explanation for the survival of oral mucosa over the osteodental acrylic lamina, whereas their absence in distant areas may have induced the oral mucosa to transdifferentiate into a conjunctival type lining. Alternatively, conjunctival regrowth from forniceal stem cells should be taken into consideration. PMID- 16371781 TI - Liquid nitrogen cryotherapy of advancing wavelike epitheliopathy. AB - PURPOSE: To describe a method for treating advancing wavelike epitheliopathy (AWE) with liquid nitrogen cryotherapy spray. METHODS: A Brymill liquid nitrogen cryotherapy canister with a "D" tip was used to freeze AWE in 5 cases in 4 patients. Freezing time was approximately 1-2 seconds and was performed twice. The affected limbus and corneal epithelium were treated in this manner. RESULTS: In each case, AWE resolved within 2 weeks, with no recurrence at 1 year. CONCLUSION: Liquid nitrogen cryotherapy is a safe and effective treatment of advancing wavelike epithelopathy. PMID- 16371782 TI - Adenoviral keratoconjunctivitis in a tertiary care eye clinic. AB - PURPOSE: To elucidate the natural history and epidemiology of adenoviral keratoconjunctivitis as seen in a tertiary care eye clinic. METHODS: In a retrospective observational case series design, the medical records of 54 consecutive patients seen over a 6-year period at the Dean McGee Eye Institute (DMEI) with a diagnosis of adenoviral keratoconjunctivitis were analyzed retrospectively. The main outcome measure was the duration of symptomatic subepithelial corneal infiltrates following onset of acute conjunctivitis. RESULTS: Annual case numbers ranged from 3 each in 1997 and 1998 to 26 in 2000. No more than 5 new cases presented in any single month, and no seasonal predominance was evident. The median duration of disease was 5 weeks, but the mean was 23.4 weeks, reflecting a subset of patients with a prolonged course. Indeed, 14 (25.9%) of 54 patients demonstrated symptomatic subepithelial corneal infiltrates for more than 45 days from the first examination for conjunctivitis. CONCLUSION: These data suggest a significant level of long-term morbidity for adenoviral keratoconjunctivitis. PMID- 16371783 TI - Quantitative anatomical differences in central corneal thickness values determined with scanning-slit corneal topography and noncontact specular microscopy. AB - PURPOSE: This study was designed to analyze the differences in central corneal thickness values determined with noncontact specular microscopy and scanning-slit corneal topography. The measurements were performed on the same eye. METHODS: We analyzed the central corneal thickness values of 93 patients (n = 93) by means of noncontact specular microscopy (Topcon SP-2000P noncontact specular microscope, Topcon Corp., Tokyo, Japan) and scanning-slit corneal topography (Orbscan Topography System II, Orbscan Inc., Salt Lake City, UT). One experienced physician performed 3 consecutive central corneal thickness measurements with both devices. RESULTS: The central corneal thickness values obtained by means of Orbscan pachymetry were 17 +/- 2.7 (range, 12-24) microm greater. A significant correlation was observed between scanning-slit corneal topography and noncontact specular microscopy (Pearson correlation coefficient, r = 0.976; P < 0.001). CONCLUSIONS: Researchers should know of the existence of this difference between noncontact specular microscopy and Orbscan pachymetry when interpreting central corneal thickness values. PMID- 16371784 TI - The use of the Fugo Blade in corneal surgery: a preliminary animal study. AB - PURPOSE: To evaluate the performance of the Fugo Blade in cutting corneal tissue in animals, both in vitro and in vivo. METHODS: Corneal incisions were made on pig eyes using the Fugo Blade. The histologic characteristics of these cuts were then evaluated. Similar cuts were also performed in rabbit corneas in vivo and compared with cuts performed with mechanical blades. The rabbits were euthanized, and the corneas were obtained for histopathologic examination 14 days after surgery. RESULTS: Slit-lamp examinations and histologic findings showed that the Fugo Blade can produce sharp cuts in the cornea with minimal tissue damage beyond the edges of the incisions. It cuts without resistance and permits normal wound healing. CONCLUSIONS: Although the Fugo Blade is currently approved by the Food and Drug Administration for anterior capsulotomy, our study demonstrates that the Fugo Blade can safely and quickly produce resistance-free cuts in corneal tissue in animals, opening additional avenues for use of this device in corneal surgery. PMID- 16371785 TI - Intraocular concentrations of gatifloxacin and moxifloxacin in the anterior chamber via diffusion through the cornea using collagen shields. AB - PURPOSE: To evaluate the penetration of gatifloxacin and moxifloxacin into the anterior chamber, and any adverse reaction to the cornea, using collagen shields presoaked in oversaturated solutions of the antibiotics. METHODS: Collagen shields, presoaked for 10 minutes in an oversaturated solution of gatifloxacin or moxifloxacin, were placed on the surface of each of the corneas of 15 rabbits for a total of 30 eyes (15 in each group). The antibiotics were prepared by dissolving the powder form of the antibiotics in a solution until no further particulate could be further dissolved. Aqueous humor samples were taken 3.5 and 6 hours later. RESULTS: The initial concentrations of gatifloxacin and moxifloxacin were 5.43 +/- 0.16 mg/mL and 3.14 +/- 0.22 mg/mL, respectively. The average concentration of gatifloxacin in the anterior chamber was higher than that of moxifloxacin at the 3.5-hour sample (6.32 +/- 2.67 microg/mL versus 3.55 +/- 3.5 microg/mL, P = 0.0034). The concentrations of both antibiotics, although decreased, remained relatively high at the 6-hour sample (1.39 +/- 1.13 microg/mL versus 0.816 +/- 0.6 microg/mL at 6 hours, respectively, P = 0.22). No obvious clinical or histologic signs of toxicity were noticed in either group. CONCLUSION: Gatifloxacin and moxifloxacin showed good penetration into the anterior chamber with no obvious adverse reaction to the cornea. The concentrations in the anterior chamber exceeded the minimal inhibitory concentration (MIC) 90 of most organisms responsible for postoperative endophthalmitis (POE). PMID- 16371786 TI - Oxygen transmissibility of piggyback systems with conventional soft and silicone hydrogel contact lenses. AB - PURPOSE: To investigate the apparent oxygen transmissibility of various piggyback systems using conventional and silicone hydrogel soft contact lenses of different water content and permeability, rigid poly(methyl methacrylate), and rigid gas permeable lenses of medium, high, and ultrahigh oxygen permeability. The aim of the study was to establish which material (rigid or hydrogel) is more representative of the resulting oxygen performance of piggyback systems. METHODS: The apparent oxygen transmissibility of 66 piggyback systems was measured with an electrochemical method. Eighteen of these combinations involved the use of silicone hydrogel contact lenses currently available. One hyperpermeable rigid gas-permeable contact lens (tisilfocon A) was also included in the study. RESULTS: Measured apparent transmissibility correlates with rigid lens permeability (r = 0.403; SE = +/-3.03 barrer/cm; P < 0.001) and hydrogel lens permeability (r = 0.334; SE = +/-3.2 barrer/cm; P < 0.001). As expected, a linear model comprising permeability values from both rigid and soft materials gave a more precise estimation of the piggyback transmissibility (r = 0.736; SE = +/ 2.02 barrer/cm; P < 0.001). The highest values of apparent oxygen transmissibility were found for the combination of tisilfocon A rigid material with any of the 3 silicone hydrogel lenses. Tisilfocon A material significantly improved the transmissibility of all piggyback systems even when conventional hydrogels are involved. CONCLUSION: The combination of hypertransmissible rigid gas permeable lenses with silicone hydrogel soft materials should result in normal corneal function under daily wear conditions. When fitting piggyback systems, clinicians must be aware of material selection to optimize oxygen performance. This is of particular importance in already compromised corneas. PMID- 16371787 TI - Mitomycin C inhibits recurrent Avellino dystrophy after phototherapeutic keratectomy. AB - PURPOSE: To report 4 patients with Avellino corneal dystrophy (ACD) who were treated with 0.02% mitomycin C (MMC) during phototherapeutic keratectomy (PTK). METHODS: ACD was confirmed by polymorphism of the exon region in the BIGH3 gene using polymerase chain reaction followed by DNA sequencing analysis. Three patients with recurrent ACD and 1 patient with primary ACD underwent PTK. Intraoperative 0.02% MMC was applied topically with a soaked microsponge. RESULTS: There was no recurrence of ACD in the heterozygotes, but there were recurrences with best-corrected visual acuity of 20/30 in the 2 homozygotes. CONCLUSION: Intraoperative use of topical 0.02% MMC in conjunction with PTK may prevent or delay the recurrence of ACD for at least for 1 year. PMID- 16371788 TI - Three cases of corneal melting after instillation of a new nonsteroidal anti inflammatory drug. AB - PURPOSE: Three cases of corneal melting caused by a new nonsteroidal anti inflammatory drug (NSAID), bromfenac sodium, are reported. CASE REPORTS: Case 1: A 58-year-old man with a history of bullous keratopathy caused by Fuchs' corneal dystrophy was treated for episcleritis with topical bromfenac sodium. After 15 days of treatment, melting (80% depth) was observed inferiorly in the paracentral cornea. Case 2: A 71-year-old man underwent uncomplicated pterygium surgery, followed by treatment with topical bromfenac sodium. After 40 days of treatment, a 60%-depth corneal melt occurred in the nasal limbus. Case 3: A 76-year-old woman had a suspected bacterial corneal ulcer that resolved with topical ofloxacin; however, after 5 days of treatment with topical bromfenac sodium, a perforation occurred in the inferonasal cornea. RESULTS: In all three cases, severe corneal melting was characterized by mild hyperemia, very faint infiltration, and mild pain. Conservative treatment, including the use of a bandage soft contact lens and/or antibiotics and lubrication, led to resolution in all cases. CONCLUSIONS: A new NSAID, bromfenac sodium, can lead to severe corneal melting. These findings, together with similar previous reports concerning diclofenac sodium and ketorolac, suggest that careful observation is required when using topical NSAIDs in the treatment of corneal disease. PMID- 16371789 TI - Corneal injury by anti-misting agent in swim goggles: a case report. AB - PURPOSE: To report a case of corneal injury by swim goggle anti-misting agent. METHODS: Case report. RESULTS: A 47-year-old man presented with ocular injury caused by swim goggle anti-misting agent. Two weeks previously, the patient felt sudden ocular pain and grit after pool water flooded his goggles while swimming. Before the patient's swim, a copious amount of anti-misting agent was sprayed inside his goggles. On examination, severe corneal epithelial defect and stroma edema of the right eye were noted. Best-corrected visual acuity (BCVA) was 20/40. Right eye corneal thickness was 625 microm. Topical corticosteroids, antibiotic eye drops, and oral tetracycline were started. After 1 week of treatment, the epithelial defect of his right cornea improved slightly. Stromal edema decreased mildly. Epithelial defect persisted at 1 month follow-up. Right eye BCVA remained 20/40. Three months after treatment, the right eye epithelial defect healed completely. Mild diffuse superficial stromal opacity was noted. BCVA improved to 20/20. At 1-year follow-up, right eye corneal thickness was 547 microm, which was thinner than the left eye. Right eye corneal endothelial cell density measured approximately 1500 cells/mm2, which was less than the left eye at 2300 cells/mm2. The faint, diffuse stromal opacity disappeared; however, white subepithelial plaques were noted in the cornea centers. After 1 month of treatment with topical corticosteroids, the plaques subsided completely. The corneas remained clear at 2 year follow-up. CONCLUSION: Anti-misting agent can cause severe corneal injuries, including persistent epithelial defects, stromal edema and opacity, endothelial cell loss, and subepithelial plaque. PMID- 16371790 TI - Corneal ulceration and perforation with ketorolac tromethamine (Acular) use after PRK. AB - PURPOSE: To report a case of corneal ulceration and perforation after PRK connected with high doses of ketorolac tromethamine (Acula). METHODS: A 31-year old man presented 5 days after PRK in the left eye with corneal ulceration and perforation requiring penetrating keratoplasty. The patient admitted to using Acular every hour, ciprofloxocin every hour, and prednisolone acetate 1% QID postoperatively. RESULTS: Laboratory tests, including corneal cultures, were normal. A diagnosis of corneal ulceration secondary to incorrect use of high-dose ketorolac tromethamine was made. CONCLUSION: Judicious patient counseling is recommended when using topical NSAIDs in the setting of PRK. PMID- 16371791 TI - Cutting and pasting corneas: combination of a corneal allograft with relocation of a crescent of autologous corneal tissue in therapeutic penetrating keratoplasty. AB - PURPOSE: A therapeutic penetrating keratoplasty (TPKP) technique that achieves satisfactory visual rehabilitation with a minimal risk of rejection is described. METHODS: A patient presented with a central corneal perforation in the left eye extending to the temporal limbus. We performed a novel surgical procedure, in which a corneal allograft was combined with relocation of a crescent of autologous corneal tissue. RESULTS: The TPKP placed the optical zone of the donor cornea over the center of the entrance pupil. The chances of an immunologic rejection were minimized by intercalating a crescent of autologous tissue between the allograft and the limbal vessels. CONCLUSION: The combination of an allograft with a crescent of autologous corneal tissue minimizes the disadvantages associated with eccentric or oversized trephination. PMID- 16371792 TI - Rhodococcus ruber as a cause of keratitis. AB - PURPOSE: To report a case of suppurative keratitis caused by Rhodococcus ruber in a healthy 31-year-old male patient. METHODS: Ocular examination of the patient revealed a central corneal epithelial defect following trauma, with severe anterior chamber reaction hypopyon. Corneal scrapings were performed from the base of the ulcer and were subjected to routine microbiologic investigation including direct microscopic techniques (Gram stain and 10% KOH), and culture was done on blood agar and potato dextrose agar. Biochemical analysis and antibiotic sensitivity testing were done on the isolate, and additional confirmation was done by DNA sequencing. RESULTS: The isolate was identified as Rhodococcus ruber, an uncommon cause of suppurative keratitis. The isolate was found to be sensitive to augmentin, erythromycin, co-trimoxazole, tobramycin, amikacin, ciprofloxacin, ofloxacin, and sulfamethoxazole. CONCLUSION: We present a case of Rhodococcus ruber, which was successfully treated. Even if the clinical picture of ulcer is nonspecific, the need of a routine microbiologic examination and sequencing proves to be confirmatory in cases of suppurative keratitis, where the causative organisms are varied in nature. PMID- 16371793 TI - Intravitreal triamcinolone acetonide for persisting cystoid macular edema after penetrating keratoplasty. AB - PURPOSE: The intravitreal application of triamcinolone acetonide as treatment of long-standing, therapy-resistant cystoid macular edema after penetrating keratoplasty is reported. METHODS: A 44-year-old patient showed therapy-resistant cystoid macular edema for 2 years after repeated penetrating keratoplasty was performed as treatment of keratoconus. The crystalline lens was clear. The patient received an intravitreal injection of approximately 20 mg of triamcinolone acetonide. Preinjection visual acuity measured 20/80. RESULTS: Within the first 4 weeks after the injection, dense cataract developed necessitating cataract surgery. After phakoemulsification, visual acuity improved to 20/50. Optical coherent tomography and fluorescein angiography showed an almost complete resolution of cystoid macular edema. Ten months after the injection, visual acuity remained at 20/50, and intraocular pressure measured 15 mm Hg without antiglaucomatous therapy taken. CONCLUSIONS: Intravitreal injection of triamcinolone acetonide may be an additional tool in the treatment of therapy resistant cystoid macular edema after penetrating keratoplasty. After intravitreal injection of triamcinolone acetonide, cataract may rapidly develop in eyes that have been intensively treated, topically and systemically, by corticosteroids for several years. PMID- 16371794 TI - Voriconazole in the management of Alternaria keratitis. AB - PURPOSE: To discuss the role of voriconazole in the management of Alternaria keratitis. METHODS: Case report and literature review. RESULTS: A 69-year-old man with a history of corneal foreign body removal developed a stromal infiltrate 2 months later that did not improve despite topical antibiotics and natamycin. On our initial examination, visual acuity (VA) was 20/40, and he had a central, full thickness, reticular appearing infiltrate. Oral clarithromycin was added because of the atypical pattern of the infiltrate. However, no improvement was noted. A repeat culture revealed coagulase-negative Staphylococcus. VA decreased to 20/200, and increased inflammation was noted a month later; a trial of topical steroids was added. After an initial improvement, he returned with progression. Repeat culture revealed Alternaria species, and topical amphotericin was started. When there was no response, he was admitted and switched to oral and topical voriconazole. Steady resolution was noted within 10 days of therapy. CONCLUSION: Suspicion must be maintained for unusual causes of infectious keratitis. Fungal infection can be difficult to eradicate even with traditional antifungals. Although not approved for ophthalmic use yet, voriconazole provided improvement with Alternaria keratitis unresponsive to amphotericin. PMID- 16371795 TI - Successful management of aniridic ocular surface disease with long-term bandage contact lens wear. AB - PURPOSE: To present an aniridic patient with failed penetrating keratoplasties OU (PK) who underwent successful repeat PK OD, which has remained clear for 4 years with continuous bandage lens (BCL) wear. METHODS: A 73-year-old aniridic woman who had a failed PK complicated by ocular surface disease refused limbal stem cell transplantation and underwent repeat PK in her right eye. The immediate postoperative course was uneventful except for mild irregularity of the graft epithelium. Two months postoperatively, she presented with an epithelial defect along the inferotemporal graft margin adjacent to an exposed, loose interrupted suture. The suture was removed, and although the initial epithelial defect healed, she suffered recurrent episodes of epithelial breakdown and generalized whorl epitheliopathy. A BCL was inserted and rendered prompt healing of the graft epithelium, which has remained smooth and intact for 4 years. RESULTS: The graft remains thin and clear with a smooth and intact epithelial surface beneath the BCL and no signs of limbal stem-cell deficiency. CONCLUSION: Long-term BCL wear provided good corneal transplant surface protection in an aniridic patient with recurrent epithelial breakdown, thereby sparing the need for limbal stem cell transplantation and the use of potentially toxic systemic immunosuppressive therapy. Meticulous follow-up is required to monitor contact lens loss and epithelial breakdown as well as signs of secondary infectious keratitis. PMID- 16371796 TI - Bilateral infectious ulcers associated with atopic keratoconjunctivitis. AB - PURPOSE: A case of atopic keratoconjunctivitis complicated by bilateral infectious ulcers is described. METHODS: Both eyes underwent complete ophthalmologic examination and in vivo confocal microscopy imaging by using the Heidelberg Retina Tomograph II Rostock-cornea-module (HRT II-RCM) and were photographed. RESULTS: Topical antibiotics and corticosteroids associated with oral antibiotics and corticosteroids were given. After 4 weeks of treatment, the corneal lesions healed completely. CONCLUSION: Bilateral infectious ulcers with perforation are a rare and severe complication of atopic keratoconjunctivitis. HRT II-RCM in vivo confocal microscopy imaging is useful in monitoring the progression of ocular surface diseases. PMID- 16371797 TI - Evaluation of nailfold videocapillaroscopic abnormalities in patients with systemic lupus erythematosus. AB - OBJECTIVE: Nailfold capillaroscopy (NFC) has been shown to have a remarkable value in the differential diagnosis of connective tissue diseases. In fact, NFC patterns reflect the microvascular changes that may play a significant role in pathogenesis. The aims of this study were to determine in patients with systemic lupus erythematosus (SLE) the prevalence of NFC patterns, to evaluate any association with clinical features and laboratory parameters. MATERIALS AND METHODS: One hundred twenty-three patients with SLE were included in this retrospective study. Video NFC parameters were analyzed in each patient. In all cases, the following parameters were evaluated: capillary arrangement, density, size, and shape. RESULTS: In patients with SLE, major capillary abnormalities were frequently observed (44 of 123 = 35.8%); however, no specific pattern was noted. There was a significant correlation between the SLEDAI index and the severity of capillary abnormalities (P < 0.0001). Pathologic capillary abnormalities were also significantly increased in SLE with positive anti-U1-RNP antibodies (P < 0.05). CONCLUSION: NFC may be a useful method to evaluate the microvascular changes in patients with SLE, and the presence of major capillary abnormalities seems to herald a more severe clinical course of the illness. PMID- 16371798 TI - Comparison of intraarticular injection of depot corticosteroid and hyaluronic acid for treatment of degenerative trapeziometacarpal joints. AB - BACKGROUND: Trapeziometacarpal joint arthritis is a disabling condition presenting with pain at the base of the thumb causing impairment of hand function. Nonoperative treatment at an early stage includes intraarticular steroid injection. Although this treatment may bring about prompt symptomatic relief, its efficacy is unpredictable. OBJECTIVES: There is previous evidence that injection of sodium hyaluronate is effective and safe in the treatment of knee arthritis. We proposed that intraarticular injection of sodium hyaluronate, for the symptomatic treatment of trapeziometacarpal joint arthritis, could provide symptomatic relief without the adverse effects of steroids. METHODS: Fifty-two patients with trapeziometacarpal joint grade II arthritis were randomized prospectively either for methylprednisolone or hyaluronate intraarticular injections. Initial evaluation included an estimation of pain, grip, pinch strengths and the functional Purdue Pegboard Test (PPT). This evaluation was repeated after 1, 3, and 6 months and statistically compared with the initial evaluation. RESULTS: In both groups, the intraarticular injection produced a relief of pain after 1 month. Grip strength improved significantly in the group treated by the steroid during the whole evaluation period. The patients treated by hyaluronate showed improvement in grip strength after 6 months and in the pinch and the PPT after 3 months. CONCLUSIONS: Steroids and hyaluronate injections were found effective in reducing pain. Hyaluronate was more effective in the improvement of some aspects of fine hand function. PMID- 16371799 TI - Effects of kinesthesia and balance exercises in knee osteoarthritis. AB - BACKGROUND: In patients with knee osteoarthritis (OA), there is a prominent loss in proprioception and kinesthesia sensation compared with control subjects of the same age and gender. OBJECTIVES: The aim of this study is the investigation of short-term clinical effects of kinesthesia and balance exercises in patients with knee OA. METHODS: This 8-week study was conducted on 66 female patients with knee OA who were randomized into 2 groups. The first group received kinesthesia and balance exercises (such as retrowalking, walking on their toes, leaning to the sides, balance board exercises, minitrampoline exercises, plyometric exercises, and so on) in addition to strengthening exercises. The second group received only strengthening exercises. RESULTS: : Statistically significant improvements were observed postexercise for both groups with respect to baseline for WOMAC, SF-36 Form, times for performing activities of daily living, isokinetic quadriceps muscle strength, and proprioceptive sensation levels. In the first group with kinesthesia training, compared with the second group, significantly greater improvements were obtained in all the subparameters that measure functional status (WOMAC-physical function value, SF-36 Form [physical function, role limitations-physical and vitality-energy or fatigue variables], 10 stairs climbing, and 10-m walking times) and in isokinetic muscle strength at high angular velocities (P < 0.05). The absolute angular error percentage (to assess proprioceptive accuracy) was significantly improved postexercise in both groups. There were no differences between the groups. CONCLUSIONS: Additive positive effects of kinesthesia and balance exercises in knee OA have been demonstrated. Used in clinical applications, they should be able to increase the functional capacities of patients. Long-term studies about efficacy and cost-effectivity of these exercises are needed. PMID- 16371800 TI - Ureteral stenosis secondary to granulomatous vasculitis. AB - Vasculitic syndromes are a group of disorders characterized by inflammation of the blood vessels. They include polyarteritis nodosa (PAN), Churg-Strauss syndrome (CS), and Wegener granulomatosis (WG) that differ on type, size, and location of the involved vessels. We report a rare case of granulomatous vasculitis leading to ureteral stenosis in the absence of other systemic manifestations of the disease. The patient was a 56-year-old woman who began to complain of left lumbar pain, nocturia, and dysuria. Computed tomography scan demonstrated a large abdominal mass with ureteral compression. A tissue specimen obtained by laparotomy revealed the presence of a chronic inflammatory process with fibrosis, granulomatous reaction, extravascular eosinophilia, and prominent vasculitis. She was put on corticosteroids and azathioprine with a decrease of the tumoral mass. This incompletely classified vasculitis was an unexpected cause of the abdominal mass. PMID- 16371801 TI - Wegener granulomatosis with massive intracerebral hemorrhage: remission of disease in response to rituximab. AB - Wegener granulomatosis (WG) is a multisystem necrotizing vasculitis that primarily involves the upper and lower respiratory tract and kidneys but can affect almost any organ, including the central nervous system (CNS). We present a patient with WG whose disease was complicated by a massive intracerebral hemorrhage (ICH) despite standard treatment with prednisone and cyclophosphomide. Although CNS involvement is not uncommon in WG, ICH is a rare complication of WG, and although the majority of patients with WG complicated by a cerebrovascular accident have a fatal outcome, our patient survived this event. The disease subsequently progressed with recurrent pulmonary involvement and renal failure. Our patient either did not tolerate or failed to respond to several immunosuppressive agents, including cyclophosphamide, methotrexate, and mycophenolate mofetil, but achieved remission after treatment with rituximab. PMID- 16371802 TI - Infliximab-induced scleredema in a patient with rheumatoid arthritis. AB - A 52-year-old patient with rheumatoid arthritis (RA) developed scleredema-like skin induration after treatment with the tumor necrosis factor alpha (TNFalpha) blocking agent, infliximab. Skin induration occurred within a few weeks of initiation of infliximab, resolved with discontinuation of the drug, and recurred with rechallenge with the drug, implicating infliximab as the offending agent. Laboratory evaluation revealed a high titer of human antichimeric antibodies (HACA). The skin induration improved within a few weeks of discontinuation of infliximab and did not recur with the use of etanercept. Scleredema has been reported in association with bacterial and viral infections, diabetes mellitus, and monoclonal gammopathies. Infliximab use should be added to the list of potential associations with scleredema. This effect appears possibly specific to infliximab and may be related to the development of HACA because it did not occur with the use of etanercept in this patient. In addition, there appears to be a complex relationship between TNFalpha and tumor growth factor beta (TGF-beta), a cytokine which promotes collagen synthesis and deposition. TNFalpha blockade with infliximab may affect TNFalpha-TGF-beta interactions and may be implicated in the development of scleredema in this case. PMID- 16371803 TI - Ochronotic arthropathy: disappearance of alkaptonuria after liver transplantation for hepatitis B-related cirrhosis. AB - The deficiency of homogentisic acid oxidase, an enzyme that is mainly found in hepatocytes, is associated with alkaptonuria and ochronosis. We report a patient with clinical and radiologic findings of ochronotic arthropathy in whom alkaptonuria disappeared and the progressive course of the disease stopped after liver transplantation for hepatitis B-related cirrhosis. PMID- 16371804 TI - Angioimmunoblastic T-cell lymphoma-associated arthritis: case report and literature review. AB - Angioimmunoblastic T-cell lymphoma (AITL) is a rare type of non-Hodgkin lymphoma with systemic manifestations, including fever, lymphadenopathy, rash, and rarely arthritis. We report the case of a patient who presented with symmetric inflammatory polyarthritis and skin nodules resembling rheumatoid arthritis (RA). The patient responded initially to low-dose prednisolone, but 12 months later, he developed typical features of AITL. The characteristics of AITL-associated arthritis from 16 additional cases from the English literature are also reviewed. AITL-associated arthritis is an uncommon manifestation of angioimmunoblastic lymphoma that can mimic RA, especially when the typical systemic features of lymphoma are absent. This type of arthritis should be included in the differential diagnosis of patients presenting with an inflammatory polyarthritis. PMID- 16371805 TI - Hereditary periodic fever syndrome sans fever or distinct periodicity presenting with psychosis. AB - The genetic basis for several hereditary periodic fever syndromes has been identified and consequently, the phenotypic spectrum of these disorders has broadened. We describe a young woman with tumor necrosis factor receptor associated periodic syndrome (TRAPS), proven by mutational analysis, who presented with psychosis but without fever, symptom periodicity, or similar family medical history. This patient represents the first case of TRAPS associated psychosis. This case illustrates the importance of mutation analysis for this group of disorders in individuals presenting with unexplained inflammatory symptoms and recurrent psychoses. PMID- 16371806 TI - An uninteresting patient. PMID- 16371807 TI - Neuropathic osteoarthropathy of the shoulder. PMID- 16371809 TI - Basic science for the clinician 35: CD1, invariant NKT (iNKT) Cells, and gammadelta T-cells. AB - As noted in previous articles in this series, the very heterogeneous MHC class I and II molecules present peptide antigens to T-cells. However, there is another family of less heterogeneous MHC-related molecules. CD1 molecules present lipid antigens, oftentimes to 2 other distinct families of T-cells: gammadelta T-cells (named because they bear a T-cell antigen receptor consisting of a gamma chain and a delta chain rather than the alpha chain/beta chain heterodimer on most T cells) and iNKT cells (T-cells that bear markers previously defined on natural killer cells). CD1 molecules and the gammadelta T-cells and iNKT cells to which they present antigens have fundamental responsibilities for immune surveillance over intracellular pathogens and serve other roles that place them at the boundary between innate and acquired immunity. The gammadelta T-cell has been implicated in the pathogenesis of many diseases, rheumatologic and otherwise, suggesting that better understanding of these cells, and of CD1 molecules, may help us explain the immunopathogenesis of some inflammatory diseases and how to craft more targeted therapies in many fields of medicine. PMID- 16371810 TI - Kikuchi-Fujimoto disease associated with systemic lupus erythematosus. PMID- 16371811 TI - Is there predilection for nonnodular rheumatoid arthritis in Arabs with HLA-DR 1? PMID- 16371812 TI - Rheumatoid arthritis and tuberculosis in the tumor necrosis factor inhibitors era: observations from Brazil. PMID- 16371813 TI - Retroperitoneal fibrosis associated with adult-onset still disease. PMID- 16371817 TI - Should videos and TV cameras be allowed at births? Pro. PMID- 16371819 TI - Depression in pregnancy: drug safety and nursing management. AB - Women who are already predisposed to depression are at increased risks during pregnancy because of endocrine changes; untreated depression in pregnant women might lead to adverse effects for both mothers and infants. This article examines outcomes associated with the use of antidepressants during pregnancy and identifies how nurses can help depressed pregnant women. It is recommended that pregnant women who have mild depression be treated with nonpharmacologic therapy, such as counseling, cognitive-behavioral therapy, or interpersonal psychotherapy. Current appropriate treatment for pregnant women with moderate and severe depression is antidepressant medication, although there is no consensus on the best antidepressants for use in pregnancy. Thus, the psychotropic drug must be chosen carefully to minimize negative effects on infants and mothers, for some studies have demonstrated deleterious effects on infants. Nurses in multiple settings who interact with pregnant women should be aware of the necessity of screening for depression. Nurses in antenatal care settings can refer appropriately screened women to mental health specialists; psychiatric nurse practitioners can identify suitable interventions based on potential risks and benefits to maternal and infant health. PMID- 16371821 TI - Evaluation of breastfeeding Web sites for patient education. AB - PURPOSE: To evaluate the quality of Web sites on breastfeeding for patient education. STUDY DESIGN AND METHOD: Descriptive study of 30 Web sites on breastfeeding for patient education, evaluated based on the Health Information Technology Institute (HITI) criteria, readability, and eight content criteria from the American Academy of Pediatrics (AAP) policy statement on breastfeeding. RESULTS: The mean Flesch-Kincaid Grade Level for readability of the 30 sites was 9.2. Seven of the sites included all eight of the content criteria from the AAP, and three sites did not include any of the information recommended by the AAP content criteria. CLINICAL IMPLICATIONS: Nurses should be able to recommend best patient education materials for their patients. The five best Web sites for breastfeeding education are identified for patient teaching, and the HITI criteria are explained for nurses to learn how to evaluate Web sites for themselves and their patients. PMID- 16371822 TI - Fathers' reflections on parenting young children with type 1 diabetes. AB - PURPOSE: To describe fathers' experiences in parenting and managing the care of their young children with type 1 diabetes. DESIGN AND METHODS: This qualitative descriptive study used 16 open-ended, in-depth interviews with 14 purposively selected, involved, educated, stably employed fathers of 15 children younger than 10 years old. The fathers were asked to describe their perspective of the initial diagnosis, how they learned diabetes care, their day-to-day care and parenting experiences, and what strategies they used in the day-to-day management. Data were managed using NVivo software and analyzed using principles of naturalistic inquiry and qualitative content analysis. RESULTS: The overarching theme was "From sadness to action," with six categories emerging: (1) "shock and awe" (after diagnosis), (2) quick response to the diagnosis and learning diabetes care: "suck it up and do it," (3) "staying in the loop" (practicing the skills, tasks, and responsibilities of diabetes management), (4) "partnerships in care" (tag teaming or co-parenting), (5) active participation (being actively involved in their children's day-to-day care and benefiting from participating in this care), and (6) mantra for living with diabetes: child first, diabetes second. CLINICAL IMPLICATIONS: Nurses working with children with diabetes can use the results of this study to better understand the role of fathers in the care of these children. Fathers underscored the importance of practicing diabetes care related tasks to improve their confidence when caring for children alone. Their practical and philosophical strategies for managing their children's chronic illness have recently been incorporated into a parent mentor intervention for children newly diagnosed with type 1 diabetes. PMID- 16371823 TI - How can we improve the nursing work environment? AB - It has been suggested by many recently that the nursing work environment needs to be altered to make it more responsive to both nurse and patient needs. One essential aspect of this change would be to increase patient safety. This article suggests that to improve patient safety as well as satisfaction of nurses, the culture of the nursing organization should be transformed into one of a "learning organization." Using this model of an organization, every member of the nursing organization would be encouraged to reach his or her greatest potential, the welfare of the team would become paramount, and a shared vision of where the organization needs to go would emerge, thus maximizing productivity, safety, and job satisfaction for all healthcare team members. This transformation could mean that the terms "Nursing Organizations" and "Learning Organizations" would not have to be oxymorons. PMID- 16371824 TI - Wonders and worries of parenting a child with Asperger syndrome & nonverbal learning disorder. AB - PURPOSE: To describe the prevalent joys and pressing concerns of parents of children with Asperger syndrome (AS) and nonverbal learning disorder (NLD). DESIGN AND METHODS: An exploratory study of written responses of 103 matched couples from survey data gathered in a larger study that examined the parental experiences of raising a child with AS and NLD. Participants were recruited from two national Internet Web sites for parents of children with AS and NLD. Data were qualitatively analyzed to develop categories and themes. RESULTS: Prevalent joys of mothers and fathers included child characteristics such as personality traits of the child, observing their child happy, and watching him/her grow, mature, and succeed. Parents' most pressing worries focused on adulthood and the future of their child. CLINICAL IMPLICATIONS: The findings of this study can inform the care given by nurses in the pediatric, family, and mental health settings. Family nurses are in a unique position to assess and respond to the special adaptive needs of families of children with learning disorders such as AS/NLD. As part of routine health screening and well-child visits, providers can ask about the joys and assess the beliefs and worries parents have; this could lead to reinforcement of their strengths and motivations and better meeting of their needs. PMID- 16371825 TI - Preserving breastfeeding for the rehospitalized infant: a clinical pathway. AB - The benefits of feeding newborns with human milk are well established. Unfortunately some hospital practices do not support successful breastfeeding; practices such as early hospital discharge after birth, lack of appropriate follow-up primary care providers, and lack of access to breastfeeding support services can contribute to breastfeeding failure, as well as morbidity and mortality in the infant. Infants experiencing breastfeeding difficulties are sometimes admitted to the hospital with diagnoses such as hyperbilirubinemia/jaundice, dehydration/hypernatremia, rule out sepsis, and weight loss/failure to thrive. This article describes a clinical pathway developed with the express purpose of maintaining and enhancing lactation in mother-infant dyads experiencing breastfeeding difficulties. The goal of the pathway is to maintain lactation and breastfeeding while returning the infant to a state of health. A key focus of the pathway is milk transfer, a concept that is missing from much of the research on lactation difficulties. The pathway considers breastfeeding from both a maternal and an infant perspective, with a goal of preserving breastfeeding. It uses technology to support the breastfeeding process and could be useful for all practitioners working with mother-infant dyads experiencing breastfeeding difficulties. PMID- 16371827 TI - Acceptance of hepatitis B vaccination by pregnant adolescents. AB - PURPOSE: To determine (1) the rate of prenatal hepatitis B virus (HBV) vaccine acceptance in HBV nonimmune pregnant adolescents, (2) if postulated behavioral and attitudinal factors are associated with HBV vaccine, and (3) the rate of actual receipt of HBV vaccine postpartum in eligible subjects. DESIGN AND METHODS: During 1999-2000, at an inner-city tertiary-care center prenatal clinic, 160 HBV nonimmune adolescents <18 years who were receiving prenatal and delivery care at the center were identified. The research nurse provided an HBV information pamphlet and offered immediate in-hospital postpartum HBV vaccination. Risk factors for nonacceptance of the vaccine were measured with structured questionnaires and medical record review documenting care attendance. Subsequently, all subjects, irrespective of prenatal acceptance of vaccine, were offered vaccine before postpartum discharge, and the rate of actual acceptance was determined. RESULTS: In these predominantly African American (95%) adolescents, the rate of vaccine acceptance was 91%. Actual vaccination rate was 86%, but it was not associated with prior acceptance of vaccination or behavioral or attitudinal factors. CLINICAL IMPLICATIONS: Acceptance of vaccination and actual vaccination were high in this population of high-risk adolescents. The authors found that reoffering vaccine was a successful intervention, even with adolescents who had less-than-optimal attendance at prenatal visits. Given the high rate of acceptance and vaccination in this setting, the authors would encourage public health programs to implement vaccination programs in adolescent prenatal clinics and to offer vaccination postpartum to those who do not receive it prenatally. PMID- 16371828 TI - Juices: when and how much. PMID- 16371829 TI - Interdisciplinary networking about patient safety. PMID- 16371830 TI - It's a small, small world. PMID- 16371837 TI - Elective preterm birth. PMID- 16371838 TI - What should a primitive surgeon know about Meniere's patients? PMID- 16371839 TI - Radiofrequency myringotomy with the topical use of mitomycin C: an experimental study. AB - OBJECTIVE: To assess the use of radiofrequency myringotomy in combination with mitomycin C as an alternative myringotomy technique on rabbits. STUDY DESIGN: Experimental animal research protocol. SETTING: University of Crete, School of Medicine, Medical Experimental Education and Research Center. METHODS: Radiofrequency myringotomies were performed under general anesthesia on both ears of 20 rabbits. Mitomycin C (0.3 mg/ml) pledgets were applied in the right ears (study group) and saline pledgets in the left ears (control group). Animals were monitored using otomicroscopy weekly until myringotomy closure. Kaplan-Meier survival techniques were used to compare myringotomy patency time between the two sides. INTERVENTION: Radiofrequency myringotomy under general anesthesia on both ears of 20 rabbits. MAIN OUTCOME MEASURE: Myringotomy patency time. RESULTS: The mean patency time of the study group was 5.45 weeks (95% confidence interval, 5.185-5.715 weeks). The mean patency rate for the control side was 1.60 weeks (95% confidence interval, 1.38-1.82 weeks). The Breslow (generalized Wilcoxon) test showed the study group to have significantly longer patency of radiofrequency myringotomy than the control group (p<0.0001). CONCLUSION: The patency period of radiofrequency myringotomy was not long enough to be proposed as an alternative to the insertion of ventilation tubes, although mitomycin C had a significant adjunct effect in prolonging the patency rate of radiofrequency myringotomy on rabbits. PMID- 16371840 TI - Caspase-3, caspase-8, and nuclear factor-kappaB expression in human cholesteatoma. AB - BACKGROUND: Cholesteatoma is characterized by the accumulation of keratinizing epithelium resulting from the proliferation and differentiation of epithelium. Researchers are presently unraveling the role that apoptosis plays in the disease seen in cholesteatoma epithelium. Caspases play a key role in apoptosis. Caspase 8, which is activated by the induction of tumor necrosis factor-alpha, leads to activation of caspase-3, which activates apoptotic nucleases. Nuclear factor kappaB is a transcription factor known to inhibit apoptosis induced by tumor necrosis factor-alpha. HYPOTHESIS: In this study, we hypothesized that expression of caspase-3, caspase-8, and nuclear factor-kappaB is uniquely connected to the proliferation, differentiation, and programmed cell death of keratinocytes during the growth and development of cholesteatoma. METHODS: We obtained 41 cholesteatoma specimens for this study. The presence of these proteins in cholesteatoma was examined using immunohistochemistry and a colorimetric assay system. We also examined the patterns of apoptosis by using terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining. RESULTS: Using the immunoperoxidase staining method, caspase-3 was found to be densely localized in the spinous and granular layers of cholesteatoma epithelium; caspase 8 was also found in the granular layer. Nuclear factor-kappaB was localized densely in the perinuclear region of the epithelium. The results obtained with immunoperoxidase staining agreed with those obtained with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining. In addition, the colorimetric assay method was used to measure the activity of caspase-3. CONCLUSION: These findings suggest that caspase-3 and caspase-8 play important roles in programmed cell death, which results in the accumulation of keratin debris during the growth of cholesteatoma. Nuclear factor-kappaB was found in cholesteatoma epithelium, but the transcription factor appeared to be inactivated. PMID- 16371841 TI - Endoscopic transtympanic tympanoplasty in the treatment of conductive hearing loss: early results. AB - OBJECTIVE: We describe our initial experience with endoscopic transtympanic tympanoplasty and evaluate whether this approach is adequate and minimally invasive in the treatment of conductive hearing loss. STUDY DESIGN: Prospective trial. SETTING: University hospital. PATIENTS: Nine patients underwent endoscopic transtympanic tympanoplasty, with an average follow-up period of 17 months. Presurgical diagnosis was made by transtympanic endoscopy through a perforation made by OtoScan laser-assisted myringotomy in the outpatient clinic. METHODS: With clean endoscopic visualization, ossiculoplasty was performed by inserting a trimmed tragal cartilage through the myringotomy perforation made by laser assisted myringotomy. Two types of ossiculoplasty were performed: columella reconstruction and interposition. The tympanic membrane was covered with a chitin membrane or sealed with a small piece of perichondrium from the tragal cartilage. MAIN OUTCOME MEASURES: Perioperative and postoperative complications and preoperative and postoperative hearing. RESULTS: Endoscopic transtympanic tympanoplasty with columella and endoscopic transtympanic tympanoplasty with interposition were performed in seven and two patients, respectively. Insertion of the cartilage was performed without conversion to a conventional otomicroscopic technique. The average hearing level before the operation was 59 dB. After the endoscopic transtympanic tympanoplasty, the average improved to the level of 27 dB, with an average air-bone gap of 11 dB. The myringotomy perforation was closed within 2 to 3 weeks. CONCLUSION: As opposed to conventional methods, this procedure does not require surgical exposure such as otosclerosis drilling and skin incision, and avoids the substantial risk of unnecessary injury to the chorda tympani. Endoscopic transtympanic tympanoplasty for a disrupted ossicular chain is an adequate and minimally invasive procedure and should prove to be a useful surgical procedure in future endoscopic tympanoplasty. PMID- 16371842 TI - Long-term results of ossiculoplasty: reasons for surgical failure. AB - OBJECTIVES: To present the value of a dedicated ear audit clinic, the overall long-term results of ossiculoplasty, and the reasons for surgical failure. STUDY DESIGN: Retrospective analysis was performed on ossiculoplasty cases in a county hospital. The information was extracted from a computer database. Individual patient case records were reviewed in all cases of surgical failure. METHODS: All patients who had ossiculoplasty were routinely followed-up in a dedicated ear audit clinic on a yearly basis. Between 1988 and 1999, the author performed 242 ossiculoplasties and their outcomes were monitored in the ear audit clinic. The 5 year results are presented. RESULTS: Even with the benefit of a dedicated audit clinic and a stable population, only 74.4% of ears had a known outcome at 5 years after ossiculoplasty. In the present series, 61.1% of partial and 37.6% of total ossicular reconstructions have an air-bone gap of 20 dB or better at 5 years. Of 83 identifiable late failures, 47 were caused by persistent or recurrent abnormalities within the middle ear and only 36 were thought to be caused by prosthesis-related or surgeon-related problems. CONCLUSION: The overall long-term results of ossiculoplasty have not dramatically improved in recent years, in spite of all the great advances in biomedical technologies. Unless there is a breakthrough in the understanding and management of the underlying otitis media, the long-term outcome of ossiculoplasty is unlikely to improve dramatically. PMID- 16371843 TI - Influence of hypercalcemia in the formation of tympanosclerosis in rats. AB - HYPOTHESIS: Hypercalcemia is thought to perform an important role in the etiopathogenesis of tympanosclerosis. The purpose of the present study was to assess the influence of hypercalcemia in the development of tympanosclerosis in rats with or without acute otitis media. BACKGROUND: The pathogenesis of tympanosclerosis is still unclear. Histologically, it is characterized by a hyaline degeneration in the lamina propria of the middle ear and mastoid mucosa and may lead to a calcification process. The process of calcification is similar to that of other pathologically calcified tissues, such as vessels, heart valves, cornea, and others, which depend on calcium serum levels. METHODS: Twenty-four Wistar rats were divided into two different groups, with one group submitted to hypercalcemia status through calcium diet supplementation and the other used as a control group (normal calcium content diet). Both were submitted to induction of tympanosclerosis by inoculation of Streptococcus pneumoniae on the right middle ear only. Otomicroscopy was conducted every 15 days, and after 7 weeks of inoculation, the animals were killed and the tympanic bullae were resected to be analyzed through light and electron microscopy. RESULTS: The group submitted to hypercalcemia presented a prevalence of tympanosclerosis of 25% against 16.7% in the control group, presenting a relative risk of 1.27 (p=0.72). CONCLUSION: The results suggest that hypercalcemia can have an influence in the development of tympanosclerosis. PMID- 16371844 TI - Titanium mesh for functional reconstruction of the mastoid cortex after mastoidectomy. AB - OBJECTIVE: To describe a rapid and effective means of reconstructing the mastoid cortex after mastoidectomy for chronic otitis media with and without cholesteatoma. STUDY DESIGN: Retrospective. SETTING: Tertiary referral otology clinic. PATIENTS: Those undergoing tympanoplasty and mastoidectomy for chronic otitis media. INTERVENTION: After tympanoplasty and mastoidectomy, the mastoidectomy defect was reconstructed using titanium mesh with subsequent closure as routine. MAIN OUTCOME MEASURES: Evaluation of soft-tissue ingrowth into the mastoid 12 to 24 months after surgery and clinical evaluation of the postauricular surgical site for accentuation of the postauricular sulcus. RESULTS: In 99 patients, the mastoidectomy defect was reconstructed using titanium mesh. None was found to have pitting of the postauricular sulcus. One patient had partial extrusion of the mesh through the ear canal. No instance of wound infection was observed. Soft-tissue growth into the mastoid cavity was found to be minimized. CONCLUSION: Titanium mesh is a rapid and effective means of minimizing soft-tissue ingrowth into the mastoid and preventing accentuation of the postauricular sulcus after mastoid surgery. PMID- 16371845 TI - A 5-year prospective study of diabetes and hearing loss in a veteran population. AB - HYPOTHESIS: Veterans with diabetes will have significantly greater hearing loss than nondiabetic veterans. BACKGROUND: The association between diabetes and hearing loss remains unclear despite the volume of research that has been devoted to the question. Often, differences in hearing thresholds between diabetic and nondiabetic patients are confounded by age and noise exposure. METHODS: In this 5 year prospective study, 342 diabetic veterans and 352 nondiabetic veterans from the Portland VA Medical Center in Oregon were tested on a variety of audiometric measures, including pure-tone thresholds. RESULTS: Age and noise exposure were accounted for in the analyses. There was a trend toward greater hearing loss in diabetic patients 60 years of age and younger across the frequency range. These differences were statistically significant only in the highest frequencies tested (10, 12.5, 14, and 16 kHz). The effects of both diabetes and noise exposure on high-frequency hearing thresholds were dependent on age. For patients older than 60 years, the mean thresholds were not significantly different. CONCLUSION: These results suggest that diabetic patients 60 years old or younger may show early high-frequency hearing loss similar to early presbycusis. After age 60, difference in hearing loss between diabetic and nondiabetic patients was reduced. PMID- 16371846 TI - Good speech recognition and quality-of-life scores after cochlear implantation in patients with DFNA9. AB - OBJECTIVE: To compare audiometric and quality-of-life results in DFNA 9 patients who received a cochlear implant with cochlear implant patients with adult-onset progressive sensorineural hearing loss. STUDY DESIGN: Prospective comparative design; results were collected cross-sectionally. SETTING: Tertiary referral center. PATIENTS: Eleven DFNA 9 patients were included in the study as well as a comparative group of 39 post-lingually deafened cochlear implant subjects with adult-onset progressive sensorineural hearing loss. INTERVENTIONS: All patients received a cochlear implant. Subjects were implanted with either the Nucleus 24 M/RCS or Med-el Combi 40+ cochlear implant systems implementing the SPEAK, ACE, or CIS+ coding strategies. MEAN OUTCOME MEASURES: Speech recognition was determined by means of phonetically balanced monosyllabic word lists. The Hearing Handicap Inventory for Adults, the Glasgow Benefit Inventory, and the Scale for the Prediction of Hearing Disability in Sensorineural Hearing Loss were used to quantify the quality of life. RESULTS: The results show that the speech perception and the quality of life of the DFNA 9 patients do not differ significantly from the control group (p=0.179; p=0.56). CONCLUSION: In spite of the fact that DFNA 9 is a disease that is known to involve cochlear dendrites, cochlear implantation is a good option for treatment of deafness in DFNA 9. PMID- 16371847 TI - Conditioning pulse trains in cochlear implants: effects on loudness growth. AB - HYPOTHESIS: The addition of a high-rate (5 kpps) conditioning pulse train to the input signal of cochlear implants will result in shallower loudness growth across the dynamic range of cochlear implant patients. BACKGROUND: High-rate conditioning pulse trains have been shown to increase the dynamic range of sinusoidal stimuli for cochlear implant recipients in a manner consistent with stochastic resonance. This study further characterizes the effects of conditioning stimuli on loudness by examining the loudness growth functions for sinusoidal stimuli both with and without conditioning. METHODS: Seven post lingually deafened adults using the Clarion CII cochlear implant participated in this study. The loudness growth functions of each subject were characterized using sinusoidal stimuli, both with and without the presence of a high-rate conditioner. Loudness was measured using magnitude estimation. RESULTS: The loudness growth functions of all seven subjects demonstrate an increase in dynamic range for sinusoidal stimuli with the addition of the conditioning pulse train. Shallower loudness growth is seen across the dynamic range with the addition of a conditioner. This result was shown for loudness growth fitted to exponential, power, and cumulative gaussian functions. CONCLUSION: The addition of high-rate conditioning pulse trains to sinusoidal stimuli presented to cochlear implant recipients results in larger dynamic ranges, with more gradual increases in loudness growth across the dynamic range. This suggests that signal processing strategies incorporating conditioning may be clinically useful, requiring less compression of the input signal and leading to less distortion perceived by cochlear implant patients. PMID- 16371848 TI - Histopathology of the peripheral vestibular system after cochlear implantation in the human. AB - OBJECTIVES: The objective of this study was to describe the histology of the peripheral vestibular system in temporal bones from patients who in life had undergone cochlear implantation and to correlate the findings with previous reports of vestibular dysfunction after cochlear implantation. This is the first quantitative report of the impact of implantation on the vestibular neuronal end organ. METHODSThere were 19 temporal bones available for histologic study. Of these, 17 were suitable for the description of the morphology of the membranous labyrinth, 8 for counting Scarpa's ganglion cells, and 6 for measuring the densities of vestibular hair cells. The bones were fixed, cut, and stained according to previously published methods. Preferably, the implanted electrode was left in situ. Vestibular hair cells were counted with Nomarski's optics. RESULTS: Differences in Scarpa's ganglion cell counts and hair cell densities between the implanted and nonimplanted sides were not statistically significant. In 59% of the implanted bones, the cochlea was hydropic, and in the majority of these bones the saccule was collapsed. CONCLUSION: Cochlear implantation does not cause deafferentation of the peripheral vestibular system. Cochlear hydrops accompanied by saccular collapse is common and may cause attacks of vertigo of delayed onset, similar to Meniere's syndrome as previously reported in several clinical series. Hydrops could be caused by obstruction of endolymphatic flow in the ductus reuniens or in the hook portion of the cochlea or by damage to the lateral cochlear wall caused by implantation. PMID- 16371849 TI - Tympanic membrane healing process and biocompatibility of an innovative absorbable ventilation tube. AB - OBJECTIVE: To investigate tympanic membrane healing process and biocompatibility of pressure-equalizing (PE) tubes, made of a novel biodegradable, absorbable material, in an animal model. BACKGROUND: Myringotomy and pressure-equalizing tube insertion is a frequent otologic procedure in children. However, results may vary because of the unpredictable extrusion rate of tubes. Furthermore, tubes that are no longer required need a surgical procedure to be removed. The ideal pressure-equalizing tube should remain in place for the time selected by the otologist, with no need for subsequent removal. This objective could be met with pressure-equalizing tubes made from a self-disintegrating material. METHODS: Pressure-equalizing tubes, made of poly-bis(ethylalanate)phosphazene, were inserted in 55 ears of 28 Hartley guinea pigs, with survival times of 10, 30, and 60 days after tube insertion. In vivo reactions between the poly bis(ethylalanate)phosphazene pressure-equalizing tubes and the tympanic membrane were studied. Tympanic membranes, middle ears, and tubes were examined by scanning electron microscopy. RESULTS: There was neither infection nor inflammatory reaction to the tube in any animal. The healing process of tympanic membranes revealed neither residual perforation nor inward spread of skin epithelium into the middle ear cavity. At 30 days, 53% of the tubes had disintegrated. At 60 days, tubes were functioning in 25% of ears. CONCLUSION: These new poly-bis(ethylalanate)phosphazene pressure-equalizing tubes are promising. The healing process of tympanic membranes is excellent, with no complications. The tube disintegration rate can be controlled by varying the formulation of the polymer, obtaining tubes with predictable resorption rates, to adapt treatment to the needs of each single patient. PMID- 16371850 TI - The 360-degree maneuver for treatment of benign positional vertigo. AB - OBJECTIVE: The purpose of this study was to determine whether rotating a patient 360 degrees in the plane of the posterior semicircular canal is effective in treating classic benign paroxysmal positional vertigo. The study also compares the features of the Epley maneuver and the Semont maneuver and correlates them to the 360-degree maneuver. STUDY DESIGN: A prospective analysis of 31 patients presenting with benign paroxysmal positional vertigo who were treated using the multiaxial positioning device. A questionnaire was administered immediately after each treatment. SETTING: The study was carried out in a private practice referral clinic for benign paroxysmal positional vertigo. PATIENTS: Subjects consisted of 31 adults who ranged in age from 44 to 95 years. INTERVENTION: Thirty-one patients were treated using the 360-degree maneuver. A multiaxial positioning device was used to rotate patients completely upside down and back into the starting position in the proper plane. MAIN OUTCOME MEASURES: Subjective improvement scores, tolerability, objective nystagmus observations, and complications were recorded. RESULTS: Subjective improvement rates were 90% after one treatment; 97% were symptom-free and nystagmus-free after a maximum of three treatment sessions. Eighty-seven percent found the procedure quite tolerable from an ergonomic standpoint. CONCLUSION: The 360-degree maneuver can be effective in treating benign paroxysmal positional vertigo. Its rate of success is comparable to the rate of success of the standard Epley maneuver. On analysis, it is strikingly similar to the Epley and Semont maneuvers. PMID- 16371851 TI - Triple semicircular canal occlusion in guinea pigs with endolymphatic hydrops. AB - HYPOTHESIS: Triple semicircular canal occlusion will eliminate rotatory stimulation to the vestibular peripheral system (as it blocks endolymphatic fluid movement) and therefore release rotatory vertigo attack. This surgery is safe in ears with endolymphatic hydrops. BACKGROUND: Semicircular canal occlusion has been used as an alternative treatment of intractable benign paroxysmal positional vertigo with varied success. Triple semicircular canal occlusion in animal models blocks the responses of the semicircular canals to rotation and spares cochleae and the otolithic apparatus. This result suggests that triple semicircular canal occlusion is a prospective method in vertigo management for patients with Meniere's disease. However, the effectiveness and safety of triple semicircular canal occlusion has not been fully evaluated in ears with endolymphatic hydrops. METHODS: Endolymphatic hydrops was established in 20 guinea pigs by endolymphatic sac obliteration. Triple semicircular canal occlusion was performed in 12 of them 120 days after endolymphatic hydrops surgery, whereas 8 others were killed for morphologic observation to confirm endolymphatic hydrops. Auditory and vestibular functions were monitored from the time before endolymphatic hydrops until 1 month after triple semicircular canal occlusion. Endolymphatic hydrops and canal occlusion were confirmed by morphologic observation. RESULTS: Successful establishment of endolymphatic hydrops was indicated by mild elevation of the auditory brainstem response threshold and tentative asymmetry in nystagmus. Endolymphatic hydrops was confirmed by cochlear morphology in all eight animals that were killed 120 days after endolymphatic hydrops surgery. After triple semicircular canal occlusion, all 12 animals showed spontaneous nystagmus with a slow component toward the side that had been operated on, head tilt, rotated walking, and tentative asymmetry in rotatory nystagmus. The static symptoms disappeared within 1 month after triple semicircular canal occlusion. Caloric nystagmus was only slightly reduced after endolymphatic hydrops as compared with the contralateral ears but could not be elicited at all after triple semicircular canal occlusion. No significant elevation in auditory brainstem response threshold was found after triple semicircular canal occlusion. The canal occlusion and endolymphatic hydrops were confirmed in all surgical ears. CONCLUSION: Triple semicircular canal occlusion is effective for eliminating the response of semicircular canals to rotation and caloric stimulation and is safe in ears with endolymphatic hydrops. Also, the static compensation to the disequilibrium is quick and complete. These results suggest that triple semicircular canal occlusion should be an option for controlling rotatory vertigo in Meniere's disease. PMID- 16371852 TI - Does patient-perceived handicap correspond to the modified clinical test for the sensory interaction on balance? AB - OBJECTIVE: To determine whether patient self-reported handicap correlates with scores obtained from the modified Clinical Test for the Sensory Interaction on Balance as assessed by the Neurocom VSR Balance Master platform. STUDY DESIGN: Prospective observational. SETTING: Balance clinic in tertiary referral center. PATIENTS: Patients referred with dizziness or imbalance as their primary complaint. OUTCOME MEASURES: The modified Clinical Test for the Sensory Interaction on Balance scores as assessed by the Neurocom VSR Balance Master platform, the Dizziness Handicap Inventory, and the Health Utilities Index Marks 2 and 3. RESULTS: One hundred fifty-nine patients were entered into the study. The mean age of participants was 54.5 years, with a female-to-male ratio of 2.1:1. The scores for the Dizziness Handicap Inventory and Health Utilities Index are similar between sexes, and although the Dizziness Handicap Inventory score did not correlate with age, Health Utilities Index 2 and 3 scores did show a negative correlation with increasing age. There are weak, positive correlations between the Dizziness Handicap Inventory and the firm surface conditions of the modified Clinical Test for the Sensory Interaction on Balance but no useful correlation with the foam conditions. Similar weak negative correlations were found between the Health Utilities Index 2 and 3 and the modified Clinical Test for the Sensory Interaction on Balance scores. CONCLUSION: Patient-perceived handicap of imbalance appears to correlate poorly with assessment of postural stability using the modified Clinical Test for the Sensory Interaction on Balance. PMID- 16371853 TI - Utricular dysfunction in patients with benign paroxysmal positional vertigo. AB - OBJECTIVE: The objective of this study was to test the hypothesis that utricular function is impaired in patients with idiopathic benign paroxysmal positional vertigo. STUDY DESIGN: Prospective cohort study. SETTING: Tertiary dizziness clinic and vestibular research laboratory. PATIENTS: Twelve patients with unilateral idiopathic benign paroxysmal positional vertigo were examined 1 week and 1 month after successful treatment with positioning maneuvers and compared with 24 healthy subjects. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Otolith function was assessed with estimation of the subjective visual vertical and analysis of the torsional otolith-ocular reflex. Unilateral stimulation of the utricle was performed on a rotator that allowed eccentric lateral displacement of the patient during earth-vertical rotation with constant velocity. The otolith ocular reflex was recorded with three-dimensional video-oculography. RESULTS: There was no difference in the estimation of the subjective visual vertical between patients and controls. The peak-to-peak amplitude of the otolith-ocular reflex torsional eye position was smaller in patients than in the control group. The gain of the unilateral otolith-ocular reflex was reduced in patients on both sides on first testing. After several weeks, only the affected labyrinth showed a reduced otolith-ocular reflex gain. CONCLUSION: Our findings document otolith dysfunction in patients with idiopathic benign paroxysmal positional vertigo possibly secondary to degeneration of the utricular macula. This finding may account for the transient mild imbalance and dizziness that some patients with benign paroxysmal positional vertigo experience even after resolution of positional vertigo. PMID- 16371854 TI - Malignant cylindroma of the external auditory canal. AB - OBJECTIVE: This report describes a case of a malignant cylindroma arising in the external auditory canal. STUDY DESIGN: Retrospective surgical case review. SETTING: Tertiary care teaching hospital. INTERVENTION: Wide local excision with margin control. RESULTS: Local control of disease at 2-year follow-up. CONCLUSION: To our knowledge, this is the first report describing a malignant degeneration of cylindroma in the external auditory canal. This tumor represents an extremely rare cutaneous neoplasm, thus raising diagnostic and therapeutic challenges. PMID- 16371855 TI - Cerebrospinal fluid leak rate after the use of BioGlue in translabyrinthine vestibular schwannoma surgery: a prospective study. AB - OBJECTIVE: To determine the effectiveness of BioGlue surgical adhesive in dural and middle ear closure after translabyrinthine vestibular schwannoma surgery. STUDY DESIGN: A prospective study. SETTING: Tertiary neurotological referral center. PATIENTS: There were 24 patients in the BioGlue series. BioGlue was used in the same manner in all cases. All patients received similar postoperative care. INTERVENTIONS: We studied the use of BioGlue and its possible effect on further reducing our department's cerebrospinal fluid leak rate for translabyrinthine vestibular schwannoma surgery. MAIN OUTCOME MEASURES: Postoperative events were documented that enabled us to determine the overall cerebrospinal fluid leak rate (including incidence of various leak routes and morbidity). RESULTS: The overall cerebrospinal fluid leak rate was 62.5% (15 of 24). Rhinorrhoea was the commonest route (80%), followed by postaural wound leak (33.3%) and external auditory canal otorrhoea (33.3%). Forty percent of cases had more than one cerebrospinal fluid leak route; 73.3% of leak cases required lumbar drain insertion, 40% needed pressure bandaging, and 66.7% had to undergo formal surgical repair. Forty percent had recurrent leaks after the initial episode had completely ceased. The mean extra stay in hospital as a result of the cerebrospinal fluid leak was 13.3 days. CONCLUSION: Our preliminary prospective study of the use of BioGlue for dural and middle ear closure in translabyrinthine vestibular schwannoma surgery demonstrated poor results. The high cerebrospinal fluid leak rate associated with the unusual presentations and ensuing management difficulties in controlling these leaks lead us to recommend that BioGlue not be used in translabyrinthine vestibular schwannoma surgery. The manufacturers have noted our results and have considered adding our recommendation to the product data sheet. PMID- 16371856 TI - Unilateral hearing loss as a presenting manifestation of granulocytic sarcoma (chloroma). AB - OBJECTIVE: To present a case of acute granulocytic sarcoma of the cerebellopontine angle whose presenting symptom was sudden onset unilateral sensorineural hearing loss. STUDY DESIGN: Case report and review of the literature (MEDLINE, 1962-2005). METHODS: A 34-year-old female patient with acute myeloid leukemia on remission admitted because of sudden hearing loss in her right ear for 10 days. She had experienced occasional tinnitus, ear fullness, and dizziness for a couple of months. After confirmation of her audiometric findings with auditory brainstem responses, the patient was put on a treatment regimen for sudden hearing loss. RESULTS: On the second day of treatment, she developed ipsilateral facial paralysis, hoarseness caused by ipsilateral vocal fold paralysis, and nystagmus. Magnetic resonance imaging of the cranium revealed findings consistent with granulocytic sarcoma at the cerebellopontine angle, infiltrating the internal acoustic canal. As increased intracranial pressure symptoms developed subsequently, subtotal tumor resection was performed. However, the patient was lost, with Cushing's triad at the second postoperative month during postoperative chemotherapy. CONCLUSION: Although up to 40% of leukemic patients may have otologic symptoms, sudden onset of sensorineural hearing loss is very rare. The patient presented in this report is the first reported case with a granulocytic sarcoma of the cerebellopontine angle who presented with acute sensorineural hearing loss. Despite the rarity of such a case, we would like to emphasize that leukemia must be kept in mind as an etiologic factor in sensorineural hearing loss and suggest that complete blood count and temporal bone imaging be routinely obtained. PMID- 16371857 TI - Using audiometric thresholds and word recognition in a treatment study. AB - OBJECTIVES: First, to examine a possible limit on significant results imposed by a progressive floor effect for hearing threshold improvement in a treatment study. This floor effect for hearing recovery suggests that if inclusion criteria are not set sufficiently high, the superiority of a treatment group may not be detectable. Second, to examine the outcomes when using two different types of criteria for significant change in a subject's word recognition score. METHODS: Several single-number criteria (e.g., 15 percentage points) are compared with the 95% (p=0.05) criteria from the binomial critical difference table for monosyllables. Critical differences for binomial variables change depending on whether the starting value lies in the middle (near 50% correct) or at either extreme of the range of scores (0 or 100%). Different judgments of significant word recognition improvement (or decrease) using binomial versus single-value criteria are presented. DATA SOURCE: A recent treatment study of sudden sensorineural hearing loss (n=318) is used to illustrate these effects. CONCLUSION: First, there is a progressive floor effect of presenting severity that co-varies with the outcome measure hearing threshold recovery. In some designs, this may act to constrain the ability to detect a significant difference. Second, in the example data set, the use of single-value criteria for significant within-subject change in word recognition (e.g., 15 percentage points) introduced a miscategorization error rate of approximately 9% when compared with the result of the binomial 95% critical difference table. PMID- 16371858 TI - Differential lidocaine sensitivity of human voltage-gated potassium channels relevant to the auditory system. AB - HYPOTHESIS: Lidocaine may lead to an alteration in the processing of hearing as observed during tinnitus by inhibiting voltage-gated potassium channels at clinically relevant concentrations. BACKGROUND: Recent molecular evidence suggests that the voltage-gated potassium channels Kv 3.1 and Kv 1.1 play an important functional role in the auditory system. Lidocaine is known to influence the auditory system and may thus exert pharmacological effects on these human potassium channels. METHODS: Patch-clamp recordings were performed on the pharmacologic action of lidocaine on Kv 3.1 channels natively expressed in SH SY5Y cells and Kv 1.1 channels expressed in HEK 293 cells. RESULTS: Lidocaine reversibly inhibited Kv 3.1 and Kv 1.1 channels in a concentration-dependent manner. The half-maximal inhibitory concentration for conductance block was 607 micromol/L for Kv 3.1 (n=47) and 4,550 micromol/L for Kv 1.1 channels (n=56), respectively. The Hill coefficients were 0.9 and 0.8. Conductance block was voltage dependent for Kv 3.1 but not for Kv 1.1 channels. The midpoint of current activation of both channels was shifted to hyperpolarized potentials. At free plasma concentrations determined during suppression (0.5-1 mg/L; 1.75-3.5 micromol/L) or induction (>1-2 mg/L; >3.5-7 micromol/L) of tinnitus Kv 3.1 and K v1.1 channels would be suppressed by at most 1.5 to 2%. CONCLUSION: Human Kv 3.1 and Kv 1.1 channels exhibited different sensitivities to the inhibitory action of lidocaine. The small effect at clinically relevant concentrations suggests that the physiologic roles of Kv 3.1 and Kv 1.1 channels in auditory neurons seem not to be impaired during the therapeutic or diagnostic application of lidocaine in the auditory system. PMID- 16371859 TI - Rapid expansion in a previously indolent cholesterol cyst: a need for lifelong follow-up. PMID- 16371860 TI - Hearing loss associated with systemic lupus erythematosus: temporal bone histopathology. PMID- 16371861 TI - Critical appraisal of the role of Ureaplasma in the development of bronchopulmonary dysplasia with metaanalytic techniques. AB - BACKGROUND: Controversy exists over whether or not Ureaplasma colonization or infection of the respiratory tract contributes to the development of bronchopulmonary dysplasia (BPD). Because BPD is a major cause of morbidity and mortality in preterm infants and a potential therapeutic intervention with antimicrobials is possible, we sought to evaluate and critique the current medical literature and to document the reported association between Ureaplasma and BPD. METHODS: We analyzed all peer-reviewed articles and previous reviews including cross-references that reported Ureaplasma respiratory tract colonization or infection and development of BPD in neonates published from January 1966 to December 2004. Inclusion criteria included a cohort limited to all neonatal intensive care unit admissions or all colonized infants, articles that did not define a numerator and a denominator for BPD and Ureaplasma or that included patients from other reports were excluded from the analysis. We evaluated BPD at 28 postnatal days (BPD28) or 36 weeks post-menstrual age (BPD36). RESULTS: Twenty-three studies with an aggregate of 2216 infants reported BPD28, and 8 studies with 751 infants reported BPD36. Although there was significant association between Ureaplasma colonization and both BPD28 and BPD36, there was substantial heterogeneity (Q test statistic, P < 0.01). We therefore focused on describing the study characteristics associated with an increased relative proportion of BPD. The greatest contribution to effect was from the studies enrolling fewer than 100 infants. CONCLUSION: Ureaplasma colonization is associated with higher reported rates of BPD, but the greatest reported effect is seen in small studies; reporting bias may be partially responsible for this effect. PMID- 16371862 TI - Medical and economic impact of a respiratory syncytial virus outbreak in a neonatal intensive care unit. AB - BACKGROUND: Respiratory syncytial virus (RSV) causes frequent nosocomial outbreaks in general pediatric wards but is less commonly reported in neonatal intensive care units (NICUs). We investigated an outbreak of RSV infection in a NICU and its impact on health care delivery, outcomes and costs. METHODS: Retrospective chart review was performed after an RSV outbreak occurred in the NICU. A case was defined as an infant with a nasopharyngeal aspirate positive for RSV by viral culture. Nucleotide sequencing of the isolates was done to determine relatedness. Hospital bills for all RSV culture-positive infants were reviewed. RESULTS: Nine infants (mean age, 34 days; mean birth weight, 1757 g; and mean estimated gestational age 31 weeks and 5 days) were infected with RSV subgroup B during this outbreak. By nucleotide sequencing, the isolates were identical. Clinical manifestations included cough, congestion, increased oxygen requirement, apnea and respiratory failure. The 5 infants requiring intubation had a significantly lower mean birth weight (1301 g versus 2328 g, P = 0.027), mean estimated gestational age (28 weeks and 5 days versus 35 weeks and 2 days, P = 0.014) and mean weight at onset of symptoms (2093 g versus 2989 g, P = 0.049) than the 4 nonintubated infants. More than 1.15 million dollars in hospital charges were attributable to the outbreak. All infants survived. CONCLUSION: Infants in a NICU who develop cough, congestion or apnea should be tested for RSV and other common respiratory viruses during the winter respiratory season. Even in a closed NICU, nosocomial outbreaks of these viruses can occur and have a major effect on healthcare delivery, costs and outcomes. PMID- 16371863 TI - Infections by human coronavirus-NL in hospitalized children. AB - BACKGROUND: A new human coronavirus (HCoV), HCoV-NL, was recently reported for Dutch patients with acute respiratory tract infections (ARTI). Little information is available on the incidence, clinical manifestations and epidemiologic features of HCoV-NL infections. METHODS: We performed a prospective study of symptomatic (case subjects with ARTI) and asymptomatic (control subjects undergoing elective surgery) children, or=50 mm redness and/or swelling). RESULTS: 288 children were vaccinated, 145 with DTaP.IPV and 143 with Tdap, and after 48 hours examiners noted local redness >or= 50mm in 17.2 and 6.3%, respectively (P = 0.004). DTaP.IPV vaccinees initially experienced local pain (in 54%) which limited arm motion (in 37%), but symptoms largely resolved by 48 hours. Tdap vaccinees had fewer symptoms (pain in 20%, limited arm motion in 14%). Children with large reactions to DTaP.IPV more often than nonreactors had elevated preimmunization antibody concentrations to 1 or more of diphtheria, pertussis toxin or pertactin and elevated postimmunization antibody concentrations to all antigens except fimbriae. Booster responses to Tdap were reduced with the smaller antigen doses but were generally satisfactory. CONCLUSIONS: This preschool DTaP.IPV booster vaccination caused large local reactions in 1 in 5 children, with transient discomfort. With Tdap vaccine, such reactions were significantly fewer but not eliminated. A Tdap.IPV vaccine warrants study for routine use at 4-6 years of age. PMID- 16371867 TI - Longitudinal study of the prevalence and prognostic implications of oral manifestations in romanian children infected with human immunodeficiency virus type 1. AB - OBJECTIVE: We evaluated the prevalence and predictive value for human immunodeficiency virus (HIV) disease progression of oral manifestations in Romanian children. METHODS: A nonrandom sample of 238 HIV-infected children was followed prospectively between 1998 and 2001 at the Romanian-American Children's Center in Constanta, Romania. Study subjects underwent comprehensive annual oral examinations. Oral manifestations of interest, demographic data and other selected medical information were recorded at baseline and at each subsequent visit. Descriptive statistics and survival analysis methods were used in the study. A level of statistical significance of alpha = 0.05 was used. RESULTS: The study subjects' mean age was 9.9 +/- 1.2 (SD) years. The most common oral lesions were gingivitis (49%), parotid enlargement (13%) and oral candidiasis (11%). Oral candidiasis was associated with progression to acquired immunodeficiency syndrome or death (whichever occurred first) (log rank P = 0.03) and with death (log rank P < 0.001). Oral hairy leukoplakia also was associated with progression to death (log rank P = 0.001). The risk of dying was 3.43 (95% confidence interval, 1.86 6.34; P < 0.001) for children who had oral candidiasis at baseline and 4.62 (95% confidence interval, 1.67-12.77; P = 0.003) for those who had oral hairy leukoplakia. CONCLUSIONS: Oral manifestations occur commonly among HIV-infected Romanian children. Oral candidiasis and oral hairy leukoplakia were positive predictors of HIV disease progression. PMID- 16371868 TI - Genotypic human immunodeficiency virus type 1 drug resistance in highly active antiretroviral therapy-treated children in Abidjan, Cote d'Ivoire. AB - OBJECTIVE: To estimate the frequency of human immunodeficiency virus type 1 (HIV 1) displaying genotypic drug resistance in highly active antiretroviral therapy (HAART)-treated children in Abidjan. METHODS: Among the 269 HIV-1-infected children enrolled in the ANRS 1278 prospective observational cohort between October 2000 and September 2003, 115 [median age, 6.35 years (range, 1.2-15)] required treatment and received HAART for at least 6 months. Treatment consisted of 2 nucleoside analogue reverse transcriptase inhibitors associated with nelfinavir (70.5%) or efavirenz (29.5%). Plasma HIV-1 RNA and CD4+ T cell counts were determined at baseline and every 6 months thereafter. Genotypic resistance tests were performed in cases of virologic failure (viral load >or=3 log10 copies/mL) after at least 6 months of HAART. RESULTS: After a median of 10.2 months of HAART, 66% (76 of 115) of children were in virologic success. Most of these children were infected with CRF02 strains. Twenty-seven viruses displayed resistance to at least 1 antiretroviral drug (27 of 38, 71%). Thirteen, 9 and 5 children had viruses with resistance to 1, 2 or 3 of the drugs included in their regimen, respectively. Resistance to lamivudine and/or to non-nucleoside analogue reverse transcriptase inhibitors was frequent among the 38 children in virologic failure. The 90M, 46L, 88S or 54V mutations were found in 11 (38%) of the 29 children taking nelfinavir. The overall frequency of viruses showing genotypic resistance to at least 1 antiretroviral drug was 23% (27 of 115) among the treated children. CONCLUSION: These results are similar to what is generally observed in industrialized countries. Despite these encouraging results, efforts are needed to maximize the long-term efficiency of treatment and to minimize the risk of emergence of drug resistance in treated children. PMID- 16371869 TI - Diagnosed tuberculosis during the follow-up of a cohort of human immunodeficiency virus-infected children in Abidjan, Cote d'Ivoire: ANRS 1278 study. AB - INTRODUCTION: Most data on tuberculosis in human immunodeficiency virus (HIV) infected children in Africa come from hospital-based and cross-sectional studies. OBJECTIVES: To estimate the incidence of tuberculosis in HIV-infected children participating in an observational cohort. METHODS: HIV-infected children in Abidjan, Cote d'Ivoire, are followed in a prospective cohort. At enrollment, all children had a physical examination, CD4 lymphocyte counts, chest radiograph and a tuberculin test. Quarterly follow-up visits are organized. All patients with suspected tuberculosis undergo specific investigations including gastric aspiration and culture. All isolates are tested for susceptibility. RESULTS: From October 2000 to December 2003, 129 girls and 153 boys were recruited. Of children without a current or previous diagnosis of tuberculosis, 6.5% (13 of 199) had a tuberculin test result of >5 mm, compared with 17.5% of children (10 of 57) with current or previous tuberculosis (P < 0.02). Forty-eight children (17%) had a history of treated tuberculosis, and 27 children were being treated for tuberculosis at enrollment or during the first month of follow-up. Eleven children were diagnosed with tuberculosis after the first month of follow-up, and the diagnosis of mycobacterial infection was confirmed in 7 cases. Of 5 tested isolates of Mycobacterium tuberculosis, 3 were resistant to at least 1 antitubercular drug. Cumulative incidence of tuberculosis was 2060/100,000 at 12 months, 3390/100,000 at 2 years and 5930/100,000 at 3 years. The 3-year risk was 12,400/100,000 in immunocompromised children (CD4 <15%) and 3300/100,000 in other children (P < 0.0001). CONCLUSION: The risk of tuberculosis among HIV-infected children in Cote d'Ivoire is strongly associated with the degree of immunodeficiency in HIV infection. PMID- 16371870 TI - Open randomized trial comparing the immunogenicity and safety of a new measles mumps-rubella vaccine and a licensed vaccine in 12- to 24-month-old children. AB - BACKGROUND: A trivalent measles-mumps-rubella (MMR) vaccine (MMR Berna) has been developed with a new mumps component, BBM-18, to replace a previously licensed MMR vaccine containing the Rubini mumps strain. Previous studies showed Rubini to confer insufficient long term protection against mumps infection. This study compared the immunogenicity and safety of MMR Berna, which is produced entirely in human diploid cells, with those of the licensed vaccine M-M-RVax (Merck & Co.). METHODS: We vaccinated 467 subjects, 12-24 months of age, in an open, randomized (1:1), phase II, multicenter study. Antibody titers were determined for each vaccine component with a plaque neutralization test (PNT) and a commercial enzyme-linked immunosorbent assay. Solicited local and systemic reactions were recorded in subject diaries for 6 weeks after vaccination. RESULTS: Seroconversion rates 6 to 8 weeks after vaccination for measles and rubella were statistically comparable for the 2 vaccines. However, mumps seroconversion rates were highly assay dependent, with significant differences being measured with the enzyme-linked immunosorbent assay (Berna, 77.4%; Merck, 91.3%; P < 0.001) but not the PNT (Berna, 84.8%; Merck, 87.6%; P = 0.42). The overall rate of systemic reactions was lower in the MMR Berna group (36.8% versus 45.9%; P < 0.05), including a significantly lower rate of fever of >38 degrees C (37.2% versus 51.8%; P < 0.01). CONCLUSIONS: MMR Berna was statistically noninferior to M-M-RVax with respect to seroconversion rates, and the BBM-18 strain elicited a level of functional antimumps antibodies comparable to the Jeryl Lynn strain, as measured with the PNT. Overall, MMR Berna was better tolerated than the comparison vaccine, particularly with respect to the frequency of fever. PMID- 16371871 TI - Age influences the emm type distribution of pediatric group A streptococcal pharyngeal isolates. AB - BACKGROUND: emm types 12, 1, 28, 3, 4, 2 and 6 (in that order) are the types most commonly associated with uncomplicated group A streptococcal (GAS) pharyngitis in the United States, together accounting for approximately 78% of isolates. OBJECTIVE: To determine whether the distribution of common pharyngeal group A streptococcal GAS types differs at various ages throughout childhood. STUDY DESIGN: We emm typed 3356 GAS isolates collected from the United States and Canada during 3 streptococcal seasons (2000-2003). Variations in prevalence by age for the 7 most prevalent emm types and the "uncommon" category (all types accounting for <5% of the total number of isolates) were analyzed and assessed for significance by chi2. RESULTS: The proportion of uncommon isolates increased significantly with increasing age from 18% in group 1 to 37% in group 4 (P = 0.001). We found a significant decrease in the proportion of the common pharyngeal emm types, specifically emm 12 and emm 4 type isolates, with increasing age (P = 0.001 and P = 0.003, respectively); there was no significant decline in the prevalence of other common pharyngeal types (emm 1, 2, 3, 6 and 28) with increasing age. CONCLUSION: Age-related changes in emm type distribution of pharyngeal GAS are present in childhood; these changes may reflect acquisition of immunity to more common types as a consequence of exposure early in life, but this remains to be demonstrated. PMID- 16371872 TI - Rapid diagnosis of pneumococcal meningitis: implications for treatment and measuring disease burden. AB - BACKGROUND: Streptococcus pneumoniae is the leading cause of childhood pneumonia and meningitis worldwide. Isolation of this organism, however, is uncommon in resource-poor countries, in part because of extensive use of prior antibiotics. A rapid, highly sensitive immunochromatographic test (ICT) for S. pneumoniae was evaluated for the diagnosis of meningitis. METHODS: Cerebrospinal fluid (CSF) from 450 children with suspected meningitis was tested with ICT, and results were compared with CSF culture, latex agglutination test (LAT) and/or polymerase chain reaction (PCR). Serial CSF specimens from 11 patients were also evaluated for duration of positive results during effective antimicrobial therapy. FINDINGS: All 122 cases of pyogenic pneumococcal meningitis positive either by culture (N = 87) or PCR (N = 35) were positive by ICT, yielding 100% (122 of 122) sensitivity. All purulent CSF specimens from patients with meningitis caused by other bacteria by culture (N = 149) or by LAT (N = 48) or those negative by culture, LAT and LytA and thus of unknown etiology (N = 20), and normal CSF specimens (N = 104) were negative by ICT. Thus the specificity of ICT also was 100% (321 of 321), although negativity of ICT was not confirmed by PCR, if it was positive for other organisms either by culture or LAT. Serotyping of S. pneumoniae strains revealed 28 different serotypes, indicating that outcome of ICT are independent of diverse capsular serotype of pneumococcus. Antigen was detected by ICT for at least 10 days after presentation, and 1 was still positive on day 20, which was longer than for either LAT or PCR. INTERPRETATION: ICT for pneumococcal antigen in CSF is 100% sensitive and specific in diagnosing pyogenic pneumococcal meningitis and can detect approximately 30% more pneumococcal meningitis cases than with culture alone. The simplicity of the test procedure and the longevity of CSF antigen detection suggest the potential utility of ICT to estimate the true burden of pneumococcal disease, as for Haemophilus influenzae type b using data from meningitis, and to guide selection of appropriate antibiotic treatment, especially in resource-poor countries with widespread prehospital antimicrobial use. PMID- 16371873 TI - Outbreak of Ralstonia pickettii bacteremia in a neonatal intensive care unit. AB - BACKGROUND: Ralstonia pickettii is a Gram-negative bacillus commonly found in soil and moist environments; however, R. pickettii is rarely isolated from clinical specimens. In August 2001, a cluster of R. pickettii bacteremia occurred among neonatal intensive care unit (NICU) infants at a California hospital. METHODS: A case-control study was conducted to determine risk factors for infection. A case was a NICU patient with R. pickettii bacteremia. Controls were NICU infants with negative blood cultures drawn during the same time period. A detailed environmental investigation was also conducted. RESULTS: We identified 18 patients with 19 distinct episodes of R. pickettii bacteremia from July 30 through August 30, 2001. All cases had intravascular access at the time of bacteremia. Although the case-control study did not implicate any statistically significant risk factors, the most likely source of the outbreak was the heparin flush prepared in the hospital pharmacy. This is supported by the following: (1) the heparin flush was the only substance introduced directly into the bloodstream of all case infants; (2) the heparin flush was used exclusively by the NICU; and (3) no further cases were identified after the heparin flush was discontinued. Cultures of remaining heparin flush and environmental cultures from the NICU were negative for R. pickettii. CONCLUSIONS: This unusual outbreak of R. pickettii bacteremia was most likely caused by contaminated heparin flush and ended after the heparin flush was discontinued. PMID- 16371874 TI - Congenital cytomegalovirus screening. PMID- 16371875 TI - Posttransplant lymphoproliferative disease. PMID- 16371876 TI - Prophylaxis against rabies in children exposed to bats. AB - Rabies has become a disease of increasing concern. One reason is that bat variant rabies is a more common cause of human disease, with 1-2 deaths per year in the United States. Bat bites are much more difficult to document than bites from larger animals. Deaths from rabies encephalitis have remained undiagnosed until postmortem examination. Prophylaxis includes a series of 5 vaccinations during 28 days. Vaccine efficacy has been documented, even in young children. PMID- 16371878 TI - Human metapneumovirus-associated lower respiratory tract infections in korean infants and young children. AB - To define the role of human metapneumovirus (hMPV) in previously healthy Korean children, a retrospective study was done on 166 children with lower respiratory tract infections and on their stored nasal aspirates. The hMPV gene was detected by reverse transcriptase-polymerase chain reaction. Twenty-six of 166 individuals tested positive for hMPV. The clinical diagnoses of hMPV infection were pneumonia in 15 children and bronchiolitis in 11 children. PMID- 16371879 TI - Mastitis in children from birth to 17 years. AB - Twenty-two cases of mastitis were evaluated between 1995 and 2003. Nine of the children were younger than 2 months of age, and 12 were older than 8 years of age. Girls accounted for 82% of the cases. Seven of the infections were true abscesses. Pathogens included Staphylococcus aureus in 5, Gram-negative bacilli in 3, group A Streptococcus in 1 and enterococcus in 1. These data suggest that mastitis in children occurs in 2 distinct age groups, neonates and pubescent/postpubescent; however, the clinical disease is similar in both populations. PMID- 16371880 TI - Regional differences in the epidemiology of invasive pneumococcal disease in toddlers in Germany. AB - In a population-based study, regional differences in incidence, serotype distribution and resistance rates in invasive pneumococcal disease in 1-2-year old children were related to different day care attendance rates. Day-care attendance appears to be a relevant risk factor in some German states and should be considered for inclusion in the recommendations for pneumococcal vaccination of children at risk. PMID- 16371881 TI - Secondary hemophagocytic lymphohistiocytosis in Turkish children. AB - Between January 1998 and January 2005, a total of 18 children 2 weeks-72 months of age were diagnosed as having secondary hemophagocytic lymphohistiocytosis. The frequency of secondary hemophagocytic lymphohistiocytosis among total hospitalized patients during this period was 0.05% (18 of 34,250). Of the 18 patients, 8 (44.5%) had bacterial infections; cytomegalovirus and Epstein-Barr virus infections were present in 5 (28%) and 1 (5.5%), patient, respectively. Leishmaniasis was diagnosed in 2 patients (11%), and herpes simplex virus was diagnosed in 2 patients (11%). Six patients died during treatment, and 1 patient was lost to follow-up. The survival rate was 61%. PMID- 16371882 TI - Discontinuation of secondary prophylaxis for Pneumocystis pneumonia in human immunodeficiency virus-infected children treated with highly active antiretroviral therapy. AB - This study shows the long term safety of discontinuing secondary prophylaxis for Pneumocystis pneumonia in 5 human immunodeficiency virus-infected children who had recovered from a confirmed episode of Pneumocystis pneumonia, had <15% of CD4 cells at the time of starting highly active antiretroviral therapy and whose CD4 cell counts increased to >15% for >or=3 months during highly active antiretroviral therapy. PMID- 16371883 TI - Intestinal obstruction complicating Yersinia enterocolitica serotype O:21 infection in an infant. AB - Intestinal obstruction is an uncommon complication of Yersinia enterocolitica infection. We report a case of enterocolitis in an 11-month-old infant, complicated by intestinal obstruction. Y. entercolitica serotype O:21, previously reported to cause severe disease, was isolated from the patient's stool. Unusual or complicated presentations of yersiniosis may be associated with more pathogenic strains of Y. enterocolitica. PMID- 16371884 TI - Subpectoral abscess a rare group A beta-hemolytic Streptococcus infection. AB - Suppurative complications of group A beta-hemolytic Streptococcus infection can present acutely and progress rapidly if prompt treatment is not initiated. Awareness of uncommon presentations is essential. We report a patient with a rare form of group A beta-hemolytic Streptococcus disease, subpectoral abscess. PMID- 16371885 TI - Transmission of community-associated methicillin-resistant Staphylococcus aureus from breast milk in the neonatal intensive care unit. AB - Methicillin-resistant Staphylococcus aureus (MRSA) has become increasingly common in neonatal intensive care units and can lead to severe outcomes. Baby C, of a set of quadruplets, died of MRSA sepsis. The surviving siblings were colonized with MRSA. Expressed breast milk was fed to all infants; tested breast milk samples were all MRSA-positive. Pulsed field gel electrophoresis results of isolates from the infants and breast milk were indistinguishable. PMID- 16371886 TI - Catheter-related fungemia caused by Saccharomyces cerevisiae in a newborn. PMID- 16371887 TI - Induction of acute otitis media by human metapneumovirus. PMID- 16371888 TI - Biomechanical assessment of the pediatric cervical spine under bending and tensile loading. AB - STUDY DESIGN: Cadaveric head-neck complexes from pediatric donors aged 2-12 years were subjected to a test battery consisting of nondestructive flexion-extension bending, nondestructive tensile step-and-hold tests, and tensile distraction loading to failure. OBJECTIVE: To characterize the biomechanical response of the pediatric cervical spine in flexion-extension bending, and the response and tolerance of the pediatric cervical spine in tensile distraction loading. SUMMARY OF BACKGROUND DATA: Except for a 19th century study on stillborn infants, to our knowledge, there are no published data from human cadaveric studies on the response and tolerance of the pediatric cervical spine. METHODS: Ten unembalmed pediatric head-neck complexes with intact ligamentous cervical spines were sectioned from pediatric donors aged 2-12 years. Each specimen was potted at the level of T2 and subjected to quasi-static nondestructive flexion-extension bending by a pure moment applied to the neck. Following the flexion-extension testing, each specimen was preconditioned in tension and subjected to a series of nondestructive tensile step-and-hold tests followed by tensile distraction loading to failure using a mini-Bionix MTS machine (MTS Systems Corp.). RESULTS: The average +/- standard deviation rotational stiffness of the skull-C2, C2-T2, and skull-T2 spinal segments was 0.72 +/- 0.07, 0.07 +/- 0.02, and 0.04 +/- 0.01 Nm/degree, respectively. Average rotational stiffness was independent of age for the range of pediatric ages tested. In the destructive tensile tests, failure occurred at an average distraction force of 726 +/- 171 N and an average distraction displacement of 20 +/- 3 mm. The average ultimate distraction force recorded for the 2-4-year-old specimens (595 +/- 143 N) was significantly lower (P < 0.05) than the corresponding average recorded for the 6-12-year-old specimens (868 +/- 71 N). Average +/- standard deviation linear stiffness in tensile loading was 34.7 +/- 5.7 N/mm. Pediatric age did not have a statistically significant effect on the average linear stiffness. CONCLUSIONS: The current study provides valuable new information on the response and tolerance of the pediatric cervical spine to quasi-static flexion-extension and tensile distraction loading. PMID- 16371889 TI - Effects of degeneration on the biphasic material properties of human nucleus pulposus in confined compression. AB - STUDY DESIGN: The biphasic compressive material properties of normal and degenerate human nucleus pulposus tissue were measured in confined compression. OBJECTIVES: The objective of this study was to determine the effects of degeneration and age on the mechanical properties of human nucleus pulposus. SUMMARY OF BACKGROUND DATA: The nucleus pulposus exhibits swelling behavior in proportion to proteoglycan content. In shear, the nucleus exhibits both fluid like and solid-like properties, suggesting a biphasic nature. To date, biphasic compressive properties of human nucleus pulpous have not been reported. METHODS: Human nucleus pulposus samples were tested in confined compression. Isometric swelling stress and effective aggregate modulus were measured. Linear biphasic theory was used to determine the permeability of the tissue. Mechanical behavior was correlated with proteoglycan and water content. RESULTS: Degeneration produced significant decreases in swelling stress (Psw = 0.138 +/- 0.029 MPa nondegenerate, Psw = 0.037 +/- 0.038 MPa degenerate) and effective aggregate modulus (H(A)(eff) = 1.01 +/- 0.43 MPa nondegenerate, H(A)(eff) = 0.44 +/- 0.19 MPa degenerate). Both properties were inversely correlated with proteoglycan content. Permeability increased with degeneration (ka = 0.9 +/- 0.43 x 10(-15) m4/N-s nondegenerate, ka = 1.4 +/- 0.58 x 10(-15) m4/N-s degenerate). CONCLUSIONS: Swelling is the primary load-bearing mechanism in both nondegenerate and degenerate nucleus pulposus. Knowledge of the biphasic material properties of the nucleus pulposus will aid the development of new treatment strategies for disc degeneration aimed at restoring mechanical function of the intervertebral disc. PMID- 16371890 TI - Acute intracranial subdural hematoma following a lumbar CSF leak caused by spine surgery. AB - STUDY DESIGN: Case report. OBJECTIVE: To report a case of an acute intracranial subdural hematoma that formed due to cerebrospinal fluid (CSF) leak following lumbar surgery. SUMMARY OF BACKGROUND DATA: Intracranial hypotension may occur when CSF is removed from the subarachnoid space. Intracranial subdural hematoma formation has been observed following significant CSF drainage during lumbar puncture or ventricular shunt placement. However, formation has been described only twice in the literature following spine surgery. METHODS: Retrospective review of the patient's medical record and head CT imaging. RESULTS: A 55-year old woman underwent lumbar surgery for failed back syndrome. Intraoperatively, a dural tear was noted and repaired. One week later, she developed expressive aphasia, and CSF drainage from her lumbar wound was noted. A head CT revealed an acute intracranial subdural hematoma with mass effect. Evacuation of the hematoma occurred via craniotomy, and the lumbar dura was repaired intraoperatively. CONCLUSIONS: We report the rare case of an acute intracranial subdural hematoma caused by a CSF leak following lumbar surgery. This report illustrates the potential morbidity associated with CSF leaks occurring after spinal surgery. PMID- 16371892 TI - Comparison of OP-1 Putty (rhBMP-7) to iliac crest autograft for posterolateral lumbar arthrodesis: a minimum 2-year follow-up pilot study. AB - STUDY DESIGN: A prospective, randomized, controlled, multicenter clinical study. OBJECTIVE: To compare the safety and clinical and radiographic outcomes of OP-1 (BMP-7) Putty to autogenous iliac crest bone graft in a population of patients undergoing laminectomy and posterolateral fusion for symptomatic lumbar stenosis associated with degenerative spondylolisthesis. SUMMARY OF BACKGROUND DATA: Although the existing preclinical and clinical data suggest that OP-1 is able to achieve osteoinduction and clinical fusion in a variety of situations, the efficacy of this recombinant protein in a clinical spine fusion population has not been fully elucidated. This study directly compares the efficacy and safety of OP-1 putty to autograft bone for arthrodesis in patients with symptomatic stenosis in association with degenerative spondylolisthesis. METHODS: Thirty-six patients with degenerative lumbar spondylolisthesis and symptoms of neurogenic claudication underwent laminectomy, bilateral medial facetectomy, and posterolateral fusion using either iliac crest autograft or OP-1 Putty. Oswestry scores and SF-36 questionnaires were used to determine the clinical response to treatment. Independent, blinded neuroradiologists reviewed both static and dynamic radiographs to determine the fusion status. Successful fusion was declared whenthe presence of continuous bridging bone between the transverse processes was observed and less than 5 degrees of angular motion and 2 mm of translational movement was measured using digital calipers. RESULTS: Efficacy data were tabulated for 27 patients at the 24-month time point and an additional 4 patients (without evaluable 24-month results) at the 36-month time point. One patient was not evaluable for radiology, so the data reflect clinical information for 31 patients and radiology for 30 patients. Clinical success, defined as a 20% improvement in the preoperative Oswestry score, was achieved by 17 of 20 (85%) OP 1 Putty patients and 7 of 11 (64%) autograft patients. A successful posterolateral fusion was achieved in 11 of 20 (55%) OP-1 Putty patients and 4 of 10 (40%) autograft patients. SF-36 scores showed similar clinical improvement in both groups. No systemic toxicity, ectopic bone formation, recurrent stenosis, or other adverse events specifically related to the use of the OP-1 Putty implant were observed. CONCLUSION: This study represents the first clinical trial to demonstrate the safety and similarity of OP-1 Putty as a replacement for autogenous bone graft in the posterolateral fusion environment with a minimum of 2-year follow-up. OP-1 Putty was able to achieve osteoinduction leading to a radiographically solid fusion in the absence of autogenous iliac crest bone graft in 55% of the patients at 24 and 36 months. These results compare favorably to the historical fusion rates reported for uninstrumented arthrodesis in this challenging clinical scenario. PMID- 16371893 TI - Predictors and modifiers of treatment effect influencing sick leave in subacute low back pain patients. AB - STUDY DESIGN: Modifying effects in multivariate analyses of a randomized controlled trial. OBJECTIVES: To identify prognostic factors for the effect of a brief intervention ("modifiers") at a spine clinic on return to work in patients with subacute low back pain. SUMMARY OF BACKGROUND DATA: A previous study of a brief intervention showed significant reduction of sick leave, compared with usual primary healthcare treatment. Randomized controlled trials give data only on the group as an average. Identifying prognostic factors that interact with the treatment ("modifiers") may identify specific groups requiring this or other types of treatment. METHODS: A total of 457 patients who had been sick-listed 8 to 12 weeks for low back pain were randomized into an intervention group (spine clinic with medical examination, information, reassurance, encouragement to engage in physical activity, n = 237), and a control group (primary health care, n = 220). All subjects filled out questionnaires. Logistic regression and tests for interaction were used to identify prognostic factors and modifiers for return to work in the two groups, at 3 and 12 months of follow-up. RESULTS: At 3 months of follow-up, the strongest modifying effect on return to work was the perception of constant back strain when working and beliefs about reduced ability to work. At 12 months, gastrointestinal complaints were the strongest modifier for the effect of the intervention. CONCLUSION: The spine clinic intervention seems to have a main effect on work absenteeism via interacting with the concerns of being unable to work. PMID- 16371894 TI - The treatment of disc herniation-induced sciatica with infliximab: results of a randomized, controlled, 3-month follow-up study. AB - STUDY DESIGN: A randomized controlled trial. OBJECTIVES: To evaluate the efficacy of infliximab, a monoclonal antibody against tumor necrosis factor (TNF)-alpha in a randomized controlled setting. SUMMARY OF BACKGROUND DATA: Recently, we obtained encouraging results in an open-label study of infliximab in patients with disc herniation-induced sciatica. Furthermore, the results of experimental studies support the use of infliximab in sciatica. Therefore, we initiated a randomized, controlled trial (FIRST II, Finnish Infliximab Related STudy) to confirm the efficacy of a single infusion of infliximab for sciatic pain. METHODS: Inclusion criteria were unilateral moderate to severe sciatic pain with an MRI-confirmed disc herniation concordant with the symptoms and signs of radicular pain. Patients had to be candidates for discectomy, as evaluated by an independent orthopedic surgeon. Forty patients were allocated to a single intravenous infusion of either infliximab 5 mg/kg or placebo. Assessments at baseline and various time points included clinical examination with measurement of straight leg raising restriction; questionnaires related to subjective symptoms (leg and back pain by 100-mm visual analog scale, Oswestry disability); sick leaves; number of discectomies; and adverse effects possibly related to treatment. The primary endpoint was a reduction in leg pain from baseline to 12 weeks, which was analyzed using a Mann-Whitney U test and repeated-measures analysis. RESULTS: A significant reduction in leg pain was observed in both groups, with no significant difference between treatment regimens. Similar efficacy was observed between treatment groups for secondary endpoints. Seven patients in each group required surgery. No adverse effects related to treatment were encountered. CONCLUSIONS: The results of this randomized trial do not support the use of infliximab for lumbar radicular pain in patients with disc herniation-induced sciatica. PMID- 16371895 TI - Ischemic injury-specific gene expression in the rat spinal cord injury model using hypoxia-inducible system. AB - STUDY DESIGN: A spinal cord injury and in vitro neural hypoxia models were used to evaluate the hypoxia responsive gene expression. OBJECTIVES: To limit the risk of unwanted overexpression of therapeutic genes, we developed a hypoxia-inducible gene therapy system using the erythropoietin (Epo) enhancer and the RTP801 promoter. SUMMARY OF BACKGROUND DATA: Gene therapy is an emerging therapeutic technique to treat spinal cord injury. However, uncontrolled overexpression of therapeutic genes in nondisease tissues during gene therapy raises a doubt about its safety. Post-traumatic ischemia is an important factor worsening the spinal cord damage, and hypoxia could regulate the gene expressions using a hypoxia inducible promoter. METHODS: The plasmids, pEpo-SV-Luc and pRTP801-Luc, were constructed. Mouse neuroblastoma cells (N2A) were used to evaluate the hypoxia inducible gene expression in vitro.- Gene transfection and expression were allowed for 24 hours under normoxia (pO2, 152 mm Hg) or hypoxia (pO2, 7.6 mm Hg). Spinal cord injury was made using clip compression. Plasmids were injected directly into the injured spinal cord immediately following injury. The gene expression was assessed by luciferase assay. RESULTS: pEpo-SV-Luc and pRTP801-Luc showed more than three times higher gene expression in N2A cells under hypoxia than normoxia. The expression level of luciferase in the injured spinal cord was higher than in the normal spinal cord. Immunostaining demonstrated that neurons, astrocytes, and capillary endothelial cells expressed luciferase in the cytoplasm. CONCLUSIONS: The pEpo-SV-Luc and pRTP801-Luc systems are effective in that they induce gene expression specifically in neurons under the hypoxic condition and spinal cord injury. PMID- 16371896 TI - The TRP2 allele of COL9A2 is an age-dependent risk factor for the development and severity of intervertebral disc degeneration. AB - STUDY DESIGN: Low back pain (LBP) and sciatica are usually caused by degenerative disc disease (DDD). Although they are common, the etiology of these conditions is poorly understood. A large population case-control study in the Southern Chinese was performed to study genetic risk factors to DDD. OBJECTIVES: To gain a better understanding of the etiology of DDD in relation to structural defects of the intervertebral disc. SUMMARY OF BACKGROUND DATA: A Finnish study found an association between LBP and sciatica with two variants of the alpha-chains of collagen IX, encoded by the Trp2 and Trp3 alleles, representing Gln326Trp and Arg103Trp amino acid substitutions in the COL9A2 and COL9A3 genes, respectively. Trp2 was found only in affected individuals (4%), whereas Trp3 was present in both affected (24%) and unaffected (9%) individuals. Because of the low frequency of the Trp2 allele in whites, the significance and contribution of this allele to DDD are not known. Using more objective criteria to define the disease by magnetic resonance imaging (MRI), we tested these alleles for association with DDD in a large population study. METHODS: Lumbar DDD, the presence of anular tears, and disc and endplate herniations were defined by MRI in 804 Southern Chinese volunteers 18 to 55 years of age. These were correlated with the frequencies of the Trp2 and Trp3 alleles. RESULTS: The Trp2 allele was present in 20% of the population and was associated with a fourfold increase in the risk of developing anular tears at 30 to 39 years and a 2.4-fold increase in the risk of developing DDD and endplate herniations at 40 to 49 years. Affected Trp2 individuals had more severe degeneration. The Trp3 allele was absent from the Southern Chinese population. CONCLUSION: This largest-ever population study using MRI to define DDD demonstrates for the first time that the Trp2 allele is a significant risk factor for the development and severity of degeneration. The association is age- dependent as it is more prevalent in some age groups than in others. The contrasting Trp allele frequencies between the Finns and the Chinese are the first indication that the genetic risk factors for DDD varies between ethnic groups. PMID- 16371897 TI - Human anulus fibrosis and nucleus pulposus cells of the intervertebral disc: effect of degeneration and culture system on cell phenotype. AB - STUDY DESIGN: Human intervertebral disc cells were harvested from patients with adolescent idiopathic scoliosis (AIS) and from donors with degenerative disc disease. Anulus fibrosis (AF) was separated from nucleus pulposus (NP), and cells were cultured separately in two different cell culture models. OBJECTIVES: To investigate changes in gene expression of human disc cells during in vitro expansion and to determine whether cells from adolescent idiopathic scoliosis donors show different gene expression profiles compared with cells from patients operated for degenerative disc disease. SUMMARY OF BACKGROUND DATA: During in vitro expansion, cells undergo a dedifferentiation process, which is characterized by a switch in gene expression. Markers for the differentiation and dedifferentiation status of human disc cells are not yet known. Moreover, it is not known whether changes in the gene expression pattern occur during the degeneration process. METHODS: Cells from AF and NP tissues were expanded in monolayer and alginate cultures under controlled and defined conditions. Cells were then harvested, and analysis of phenotype was performed using quantitative real-time polymerase chain reaction (PCR). The mRNA expression of Type I, II, and X collagen, aggrecan, and interleukin-1beta in scoliosis and degenerative human intervertebral disc cells was analyzed. RESULTS: The gene expression of Type II and X collagen and of aggrecan significantly decreased for both cell types during monolayer expansion. Reexpression of all genes was observed when cells were cultured in alginate. Additionally, NP cells from degenerative tissues displayed significant lower levels of Type II collagen compared with NP cells from scoliosis donors. CONCLUSIONS: These results provide a better understanding of how the phenotype of human healthy and degenerative disc cells is influenced by in vitro expansion. This may be useful for future tissue engineering purposes. PMID- 16371898 TI - De novo osteogenesis from human ligamentum flavum by adenovirus-mediated bone morphogenetic protein-2 gene transfer. AB - STUDY DESIGN: In vitro and in vivo experiment using degenerated human ligamentum flavum (LF) and Type 5 adenovirus construct with bone morphogenetic protein-2 (BMP-2) cDNA. OBJECTIVES: To demonstrate in vitro and in vivo osteogenic effect of BMP-2 gene transfer to human LF and to propose genetically modified LF as a substitute for autogenous bone graft in spinal fusion. SUMMARY OF BACKGROUND DATA: Spinal fusion is still considered to be an important option for treating various spinal disorders. To induce solid spinal fusion, osteoinductive and/or osteoconductive agents have been widely adopted. Autogenous LF, however, has never been seriously considered as a carrier for ex vivo osteoinductive gene therapy for spinal fusion. METHODS: In vitro experiment: Degenerated human LF was harvested and cultured. Type 5 adenovirus lacZ (Ad/lacZ) and BMP-2 construct (Ad/BMP-2) were produced. LF cell cultures were then exposed to Ad/BMP-2. Expressions of osteocalcin and BMP-2 mRNA were analyzed by reverse transcriptase polymerase chain reaction (RT-PCR). Western blot analysis was performed to detect osteocalcin protein. Alkaline phosphatase and von Kossa stains were used to detect osteogenic markers and bone nodule formation, respectively. In vivo experiment: Human LF tissues treated with Ad/lacZ, Ad/BMP-2, and saline were implanted into the subcutaneous tissue of nude mice. After 4 weeks, nude mice were radiographed and killed. Implanted LF tissues were harvested and histologically stained. RESULTS: LF cell cultures with Ad/BMP-2 revealed strong expression of BMP-2 and osteocalcin mRNA in RT-PCR and osteocalcin protein in western blot analysis. LF cell culture with saline showed baseline expression of BMP-2, osteocalcin mRNA, and osteocalcin protein, respectively. Furthermore, LF cell culture with Ad/BMP-2 demonstrated the expression of alkaline phosphatase and bone nodule formation in the aforementioned histochemical stain. LF tissues with Ad/BMP-2 revealed de novo osteogenesis in nude mice, whereas LF with Ad/lacZ or saline showed only remaining LF tissue without sign of bone formation. CONCLUSION: Human LF cells transduced with Ad/BMP-2 exhibited the expression of osteogenic phenotype and bone nodule formation. Additionally, genetically modified human LF with BMP-2 cDNA clearly demonstrated de novo osteogenesis, which supports the concept that biologically modified LF can be a substitute for autogenous bone graft in spinal fusion surgery. PMID- 16371899 TI - Effects of charite artificial disc on the implanted and adjacent spinal segments mechanics using a hybrid testing protocol. AB - STUDY DESIGN: Finite element model of L3-S1 segment and confirmatory cadaveric testing were used to investigate the biomechanical effects of a mobile core type artificial disc (Charite artificial disc; DePuy Spine, Raynham, MA) on the lumbar spine. OBJECTIVE: To determine the effects of the Charite artificial disc across the implanted and adjacent segments. SUMMARY OF BACKGROUND DATA: Biomechanical studies of artificial discs that quantify parameters, like the load sharing and stresses, are sparse in the literature, especially for mobile-type core artificial disc designs. In addition, there is no standard protocol for studying the adjacent segmental effects of such implants. METHODS: Human osteo-ligamentous spines (L1-S1) were tested before and after L5-S1 Charite artificial disc placement. The data were used to validate further an intact 3-dimensional (3-D) nonlinear L3-S1 finite element model. The model was subjected to 400-N axial compression and 10.6 Nm of flexion/extension pure moments (load control) or pure moments that produced the overall rotation of the L3-S1 Charite model equal to the intact case (hybrid approach). Resultant motion, load, and stress parameters were analyzed at the experimental and adjacent levels. RESULTS: Finite element model validation was achieved only with the load-controlled experiments. The hybrid approach, believed to be more clinically relevant, revealed that Charite artificial disc leads to motion increases in flexion (19%) and extension (44%) at the L5-S1 level. At the instrumented level, the decrease in the facet loads was less than at the adjacent levels; the corresponding decrease being 26% at L3-L4, 25% at L4-L5, and 13.4% at L5-S1 when compared to the intact. Intradiscal pressure changes in the L4-L5 and L3-L4 segments were minimal. Shear stresses at the Charite artificial disc-L5 endplate interface were higher than those at S1 interface. However, in the load control mode, the increase in facet loads in extension was approximately 14%, as compared to the intact case. CONCLUSIONS: The hybrid testing protocol is advocated because it better reproduces clinical observations in terms of motion following surgery, using pure moments. Using this approach, we found that the Charite artificial disc placement slightly increases motion at the implanted level, with a resultant increase in facet loading when compared to the adjacent segments, while the motions and loads decrease at the adjacent levels. However, in the load control mode that we believe is not that clinically relevant, there was a large increase in motion and a corresponding increase in facet loads, as compared to the intact. PMID- 16371900 TI - Biomechanical analysis of posterior fixation techniques in a 360 degrees arthrodesis model. AB - STUDY DESIGN: A biomechanical study to assess the ability of posterior fixation techniques to stabilize a functional spine unit (FSU) after insertion of an anterior load-sharing device. OBJECTIVE: The objective of this study is to compare various posterior fixation techniques in combination with an anterior load-sharing implant. SUMMARY OF BACKGROUND DATA: Pedicle screws and translaminar facet screws have been shown to improve the stiffness of an FSU in combination with an anterior load-sharing device. No published studies, to our knowledge, have compared translaminar facet screw fixation versus bilateral and unilateral pedicle screw fixation used with an anterior load-sharing device. METHODS: Ten cadaveric FSUs were potted using methylmethacrylate and attached to a spine simulator mounted to an MTS Mini-Bionix testing machine. The simulator was configured to control compressive loading, axial torque, flexion, extension, and lateral bending. Each specimen was tested in the intact state and following the application of each of four stabilization techniques: custom cage alone, cage plus translaminar facet screw fixation, cage plus unilateral pedicle screw and plate fixation, and cage plus bilateral pedicle screw and rod fixation with transverse coupling. Compressive stiffness and total range of motion (ROM) between +/-8 Nm of torque were extracted from the raw data. RESULTS: Each fixation method decreased ROM in torsion, flexion-extension, and lateral bending compared with the intact state. Unilateral pedicle fixation offered less stability than either of the other posterior fixations in all modes of testing except axial loading, where it was equivalent. Translaminar facet screw fixation was equivalent to bilateral pedicle screws in all modes tested. CONCLUSIONS: Using a load-sharing interbody implant, translaminar facet screws are equivalent to bilateral pedicle screws in resisting motion in all three planes. Translaminar facet screws and bilateral pedicle screws offer greater stabilization in all three planes compared with unilateral pedicle screws and a single plate. PMID- 16371901 TI - Biomechanical changes at adjacent segments following anterior lumbar interbody fusion using tapered cages. AB - STUDY DESIGN: A biomechanical evaluation of anterior cages in a calf lumbar spine model. OBJECTIVES: To determine changes in spinal motion and intradiscal pressures at immediately adjacent lumbar motion segments following anterior insertion of tapered cages. SUMMARY OF BACKGROUND DATA: Stand-alone anterior lumbar interbody fusion (ALIF) is an effective approach in the treatment of discogenic low back pain. A tapered lumbar (LT) cage design attempts to restore physiologic lordosis and sagittal balance. We are not aware of any previous biomechanical evaluation of the effects of LT cages on adjacent motion segments. METHODS: Nine fresh calf spines (L2-L5) were procured for the study. Pure moments (up to 8.5 Nm) in flexion, extension, and lateral bending were applied to the L2 vertebra in five steps through a nonconstrained loading system. With each step of loading, three-dimensional rotation at three intervertebral disc levels was obtained through a three-camera motion analysis system, and intradiscal pressures within the nucleus pulposus of the two nonoperated discs were measured with miniature transducers. The spines were tested initially intact and following paired anterior LT cage insertion. RESULTS: Following ALIF, small to moderate increase in motion was found at both adjacent segments in flexion (superior: 12.5%, P < 0.05; inferior: 11.3%, P < 0.02) and lateral bending (superior: 7.8%, P < 0.02; inferior: 6.6%, P < 0.02). An increase in intradiscal pressure was noted at the superior adjacent segment under flexion (21%, P < 0.01) and lateral bending (16%, P < 0.03). Intradiscal pressure changes at the inferior adjacent level were not significant. CONCLUSIONS: Statistically significant changes in intradiscal pressures and motion were found at the adjacent levels following a single-level stand-alone ALIF procedure using paired LT cages. PMID- 16371902 TI - Finger motion analysis of the patients with cervical myelopathy. AB - STUDY DESIGN: Pinch motion between the thumb and the index finger in myelopathy hand was analyzed using micro-reflective markers and the VICON system. OBJECTIVE: To clarify the disturbance in small finger motions of the myelopathy hand. SUMMARY OF BACKGROUND DATA: Specificity of hand dysfunction of the patients with cervical myelopathy was recently recognized as the myelopathy hand. The author established the video-based finger motion analysis system and have been applied it to some disease with abnormal pattern in finger movement. METHODS: The subjects included 11 normal persons (normal group; mean age, 30.2 years) and 9 patients with cervical myelopathy caused by spondylosis, ossification of the posterior longitudinal ligament, or disc herniation (myelopathy group; mean age, 56.0 years). All patients exhibited myelopathy hand, which was defined by two specific signs: the finger escape sign and the inability to grip and release rapidly with the fingers. Fourteen 3-mm micro-reflective markers were placed on the dorsal surface of the thumb and the index finger, and on the dorsal hand. Repeated pinching of a small ball, coin, and small rod was recorded by four infrared video cameras with the movement of each marker tracked in three dimensions. The flexion angle of the thumb IP and MCP joints, and the DIP, PIP, and MCP joints of the index finger were calculated as the angle between the vector joining the markers. RESULTS: When pinching the ball, the mean maximum flexion angle and arc of motion of the index DIP and PIP joints were significantly larger in the myelopathy group than in the normal group, while there was no difference in the MCP joint. In the thumb, the maximum flexion angle and the arc of motion of the thumb MCP joint were significantly larger in the myelopathy group than in the normal group. In addition, the arc of motion of the thumb IP joint was larger in the myelopathy group than in the normal group. Similar results were obtained when pinching the coin and small bar. CONCLUSION: Overall, the patients with myelopathy hand exhibited a larger flexion angle in the two distal finger joints when pinching small objects. PMID- 16371903 TI - The transmission of stress to grafted bone inside a titanium mesh cage used in anterior column reconstruction after total spondylectomy: a finite-element analysis. AB - STUDY DESIGN: A finite-element study of posterior alone or anterior/posterior combined instrumentation following total spondylectomy and replacement with a titanium mesh cage used as an anterior strut. OBJECTIVES: To compare the effect of posterior instrumentation versus anterior/posterior instrumentation on transmission of the stress to grafted bone inside a titanium mesh cage following total spondylectomy. SUMMARY OF BACKGROUND DATA: The most recent reconstruction techniques following total spondylectomy for malignant spinal tumor include a titanium mesh cage filled with autologous bone as an anterior strut. The need for additional anterior instrumentation with posterior pedicle screws and rods is controversial. Transmission of the mechanical stress to grafted bone inside a titanium mesh cage is important for fusion and remodeling. To our knowledge, there are no published reports comparing the load-sharing properties of the different reconstruction methods following total spondylectomy. METHODS: A 3 dimensional finite-element model of the reconstructed spine (T10-L4) following total spondylectomy at T12 was constructed. A Harms titanium mesh cage (DePuy Spine, Raynham, MA) was positioned as an anterior replacement, and 3 types of the reconstruction methods were compared: (1) multilevel posterior instrumentation (MPI) (i.e., posterior pedicle screws and rods at T10-L2 without anterior instrumentation); (2) MPI with anterior instrumentation (MPAI) (i.e., MPAI [Kaneda SR; DePuy Spine] at T11-L1); and (3) short posterior and anterior instrumentation (SPAI) (i.e., posterior pedicle screws and rods with anterior instrumentation at T11-L1). The mechanical energy stress distribution exerted inside the titanium mesh cage was evaluated and compared by finite-element analysis for the 3 different reconstruction methods. Simulated forces were applied to give axial compression, flexion, extension, and lateral bending. RESULTS: In flexion mode, the energy stress distribution in MPI was higher than 3.0 x 10 MPa in 73.0% of the total volume inside the titanium mesh cage, while 38.0% in MPAI, and 43.3% in SPAI. In axial compression and extension modes, there were no remarkable differences for each reconstruction method. In left-bending mode, there was little stress energy in the cancellous bone inside the titanium mesh cage in MPAI and SPAI. CONCLUSIONS: This experiment shows that from the viewpoint of stress shielding, the reconstruction method, using additional anterior instrumentation with posterior pedicle screws (MPAI and SPAI), stress shields the cancellous bone inside the titanium mesh cage to a higher degree than does the system using posterior pedicle screw fixation alone (MPI). Thus, a reconstruction method with no anterior fixation should be better at allowing stress for remodeling of the bone graft inside the titanium mesh cage. PMID- 16371904 TI - Does anterior plating of the cervical spine predispose to adjacent segment changes? AB - STUDY DESIGN: In a human cadaveric model, the effects of plate supplementation on the mechanical behaviors of adjacent segments were investigated. OBJECTIVES: The objective was to determine the effects of anterior cervical fusion and plating on the adjacent segments. SUMMARY OF BACKGROUND DATA: Increases in intradiscal pressure and intervertebral motion at adjacent segments have been reported in the lumbar spine following an instrumented fusion. It is unclear if the same phenomenon presents in the cervical spine. METHODS: Seven human cadaveric cervical spines (C2-T1) were used, and fusion of the C5-C6 segment was chosen for the purpose of this study. Two miniature pressure transducers were implanted within each adjacent disc. Flexion, extension, lateral bending, and torsion loads up to 2.5 Nm were applied to the intact spine, and following each of the two procedures, anterior discectomy and grafted fusion, and anterior plating of the C5-C6 motion segment. RESULTS: At the surgical level, a significant increase in segmental stiffness was observed after plating in all directions. Following the grafted fusion, there were no statistically significant changes at the superior adjacent segment, and there was a 13.7% increase in axial rotation in the inferior adjacent segment. Once anterior plating was applied, slight increase (<12%) over the intact spines was noted in lateral bending in both adjacent segments. However, there was no significant difference between the grafted fusion and anterior plated fusion at either adjacent segment. At both adjacent disc levels, the differences in intradiscal pressures between grafted fusion and plated fusion were less than 30% in all directions, and none of these differences was statistically significant. CONCLUSIONS: Intradiscal pressures and intervertebral motion at the adjacent levels are not significantly affected by the instrumented anterior fusion. The clinically observed degenerative change at adjacent segments in the cervical spine is more likely to be attributed to natural progression of the spondylotic process as opposed to biomechanical effect of the instrumentation or fusion. PMID- 16371906 TI - Reflex muscle contraction in the unaware occupant in whiplash injury. AB - STUDY DESIGN: Computer modeling and parametric analysis were used to determine the effect of reflex contraction of the neck muscles in the unaware occupant in whiplash. OBJECTIVE: To delineate effects of reflex contraction on spinal segmental kinematics during the retraction phase. SUMMARY OF BACKGROUND DATA: The ability of reflex neck muscle contraction to mitigate whiplash injury in the unaware occupant remains unclear. Analyzing relative timing between electromyographic and head-neck kinematics, previous investigators theorized that muscle contraction alters spinal kinematics, decreasing injury likelihood. Other investigators suggested that injury occurs during the initial (retraction) phase of head-neck kinematics, before significant muscle force generation. METHODS: Computer modeling was used to determine reflex contraction effects on segmental angulations, implementing parametric analysis techniques to vary reflex delay and impact severity. RESULTS: Shorter reflex delays had a greater effect on segmental angulations later in the event and at lower impact severities. However, the magnitude of this effect, particularly at higher impact severities and during maximum cervical S-curvature (factors implicated in the whiplash injury mechanism) was minimal, altering segmental angulations by a maximum of 19%. CONCLUSIONS: Because reflex contraction did not substantially alter spinal kinematics, muscle contraction likely does not initiate in sufficient time to mitigate whiplash injuries that may occur during the retraction phase. PMID- 16371908 TI - The clinical risk of vertebral artery injury from cervical pedicle screws inserted in degenerative vertebrae. AB - STUDY DESIGN: Postoperative outcomes of cervical pedicle screw (CPS) placement were evaluated. The screws were inserted in degenerative vertebrae using anatomic landmarks. OBJECTIVE: To evaluate the risks of this procedure caused by misplacements and subsequent complications. SUMMARY OF BACKGROUND DATA: The CPS gives superior vertebral fixation but risks causing vertebral artery and spinal cord injury. However, few clinical reports have been published concerning CPS misplacement and subsequent complications. Therefore, the risk of this procedure is still unclear. METHODS: There were 18 consecutive patients, who submitted to CPS fixation of degenerative vertebrae C2-C6, evaluated using computerized tomography (CT). In 5 of 9 patients in whom the screw critically violated the transverse foramen, viability of the vertebral artery was evaluated using multi detector row CT angiography. RESULTS: There were no intraoperative vertebral artery or spinal cord injuries and no serious postoperative complications. A total of 86 screws were inserted, and 25 (29%) breached the pedicle cortex. Of them, 84% deviated laterally and violated the transverse foramen. There were 13 screws (15%) that deviated more than 2 mm. However, multi-detector row CT angiography showed the continuity of the vertebral artery in all cases. CONCLUSIONS: The deviation rate of CPS placement using anatomic landmarks was higher than anticipated, although no serious complications were encountered. Refinement of the technique (e.g., using a computer-assisted navigation system) is strongly recommended. PMID- 16371909 TI - Balloon kyphoplasty is effective in deformity correction of osteoporotic vertebral compression fractures. AB - STUDY DESIGN: A prospective radiographic analysis of deformity correction during the balloon kyphoplasty procedure. OBJECTIVE: To determine the spontaneous reduction of the deformity in prone position, the subsequent deformity correction by the inflatable bone tamp, and the overall deformity correction after deposition of the cement. SUMMARY OF BACKGROUND DATA: Fracture mobility has been shown to contribute to fracture reduction in vertebroplasty. Spontaneous reduction has not been taken into account in recently published series of balloon kyphoplasty, but it must be considered when performing vertebral augmentation and when reporting and interpreting the significance of vertebral height restoration. METHODS: A consecutive series of 39 osteoporotic vertebral compression fractures were treated in 30 patients. Lateral radiographs were taken and analyzed at six different time points: 1) Preoperative standing. During the kyphoplasty procedure, four consecutive radiographs were obtained: 2) after placing the patient in prone position on the operation table, 3) after inflation of the bone tamp (IBT), 4) after deflation and removal of the IBT, and 5) after deposition of the cement. 6) Standing lateral radiographs were taken after the procedure. All fractures were analyzed for improvement in sagittal alignment (Cobb angle, kyphotic angle, sagittal index, vertebral height), complications, and reduction of pain (VAS). RESULTS: Placement of the patient in prone position displayed a significant spontaneous reduction in deformity of 6.5 degrees +/- 4.1 degrees Cobb angle. Inflation of the IBT demonstrated a further reduction of the fracture and a significant improvement of the Cobb angle of 3.4 degrees compared with baseline prone. After deflation and removal of the IBT and placement of the cement, no significant loss of fracture reduction was seen. Postoperative measurement of the Cobb angle by means of standing radiographs demonstrated a 3.1 degrees significant loss of reduction compared with the intraoperative measurement in prone position after cement application. Cement leaks occurred in 9 of 39 vertebral fractures. All patients subjectively reported immediate relief of their typical fracture pain. The VAS score significantly improved from 8.7 +/- 1.4 before surgery to 2.3 +/- 0.9. CONCLUSION: The restoration of height in kyphoplasty is attributed to dynamic fracture mobility as well as to the expansion of the inserted balloon tamp. PMID- 16371910 TI - Temporary external transpedicular fixation of the lumbosacral spine: a prospective, longitudinal study in 330 patients. AB - STUDY DESIGN: In this study, 330 patients with incapacitating low back pain underwent temporary external transpedicular fixation (TETF) of the lumbosacral spine in a prospective trial. OBJECTIVE: To evaluate TETF as a test for selecting suitable candidates for segmental spinal fusion. SUMMARY OF BACKGROUND DATA: Few studies regarding TETF have been published, and contradictory results concerning predictive value and morbidity were reported. METHODS: All patients were tested with the external fixator in two different positions: fixation and nonfixation. Before and during the test and at follow-up examination, pain was assessed on a Visual Analogue Scale (VAS). The TETF test was considered to be positive if the VAS score in the fixation state was 30 or more points lower than in the nonfixation state. Hence, a positive test would imply the decision to perform segmental lumbosacral fusion. When the reduction was less than 30 points, the test was negative. Individual pain reduction and working capacity were taken as measure of outcome. RESULTS: Most of the patients in this study (62%) underwent spinal surgery previously. The positive and negative TETF groups were quite similar, but a large within-group variation was found. Within the fusion group of 123 patients, improvement in VAS scores and improvement in working capacity were not significantly better for the positive TETF group in comparison with the negative TETF group. CONCLUSION: In this heterogeneous group of chronic patients with low back pain, TETF of the spine (including a placebo trial) does not appear to be of value in selecting suitable candidates for spinal fusion. PMID- 16371911 TI - Incidence and course of low back pain episodes in the general population. AB - STUDY DESIGN: Population-based, prospective cohort. OBJECTIVES: To estimate incidence and course of severity-graded low back pain (LBP) episodes in adults. SUMMARY OF BACKGROUND DATA: Past studies report variable estimates that do not differentiate LBP severity. METHODS: An incidence cohort of 318 subjects free of LBP and a course cohort of 792 prevalent cases was formed from respondents to a mailed survey. Incident, recurrent, persistent, aggravated, improved, and resolved episodes were defined by the Chronic Pain Questionnaire. The follow-up at 6 and 12 months was 74% and 62%, respectfully. Annual estimates were age and sex standardized. RESULTS: The cumulative incidence was 18.6% (95% confidence interval [CI], 14.2%-23.0%). Most LBP episodes were mild. Only 1.0% (95% CI, 0.0% 2.2%) developed intense and 0.4% (95% CI, 0.0%-1.0%) developed disabling LBP. Resolution occurred in 26.8% (95% CI, 23.7%-30.0%), and 40.2% (95% CI, 36.7% 43.8%) of episodes persisted. The severity of LBP increased for 14.2% (95% CI, 11.5%-16.8%) and improved for 36.1% (95% CI, 29.7%-42.2%). Of those that recovered, 28.7% (95% CI, 21.2%-36.2%)had a recurrence within 6months,and 82.4% of it was mild LBP. Younger subjects were less likely to have persistent LBP (incidence rate ratio, 0.88; 95% CI, 0.80-0.97) and more likely to have resolution (incidence rate ratio, 1.26; 95% CI, 1.02-1.56). CONCLUSIONS: Most new and recurrent LBP episodes are mild. Less than one third of cases resolve annually, and more than 20% recur within 6 months. LBP episodes are more recurrent and persistent in older adults. PMID- 16371912 TI - Prediction of thoracic dimensions and spine length based on individual pelvic dimensions in children and adolescents: an age-independent, individualized standard for evaluation of outcome in early onset spinal deformity. AB - STUDY DESIGN: A retrospective review of spine, chest, and pelvic dimensions of normal CT scans from 1999 to 2004 was performed. OBJECTIVE: To determine age independent predictors of spine and chest dimensions. SUMMARY OF BACKGROUND DATA: Progressive thoracic deformity is associated with early-onset spinal deformities. Patients with severe early-onset scoliosis are commonly smaller and slower to grow than normal. Newer treatment approaches emphasize spine length and chest growth. Assessing treatment outcomes is hampered by the lack of normal values appropriate for diseased children. This study establishes normal range standards of chest and spine dimensions to aid in assessment of treatment outcomes. METHODS: A total of 198 CT scans of previously healthy patients 0 to 21 years of age were measured. Measurements taken from CT scans include maximum inner chest width, anteroposterior chest depth, thoracic spine height, lumbar spine height, total spine height, and pelvic inlet width. Linear regression was applied to establish gender-specific reference ranges for spine and chest dimensions based on pelvic width. RESULTS: Chest and spine dimensions were significantly correlated with pelvic inlet width and gender (P < 0.001). Multiple linear regression indicated that pelvic inlet width provided excellent fit in establishing gender-specific normal ranges independent of age. CONCLUSION: Pelvic inlet width is an age-independent predictor of pediatric chest width and thoracic dimensions that may be used to assess thoracic and spine growth in early onset spine deformity patients. PMID- 16371913 TI - Long-term results of disc excision for recurrent lumbar disc herniation with or without posterolateral fusion. AB - STUDY DESIGN: A retrospective study assessing the long-term outcomes of repeat surgery for recurrent lumbar disc herniation. OBJECTIVES: To evaluate the results of repeat surgery for recurrent disc herniation, and compare the results of disc excision with and without posterolateral fusion. SUMMARY OF BACKGROUND DATA: The outcomes of revision surgery varied owing to the mixed patient populations. The optimal technique for treating recurrent disc herniation is controversial. METHODS: The sample included 41 patients who underwent disc excision with or without posterolateral fusion, with an average follow-up of 88.7 months (range, 60-134 months). Clinical symptoms were assessed based on the Japanese Orthopedic Association Back Scores. All medical and surgical records were examined and analyzed, including pain-free interval, intraoperative blood loss, length of surgery, and postsurgery hospital stay. RESULTS: Clinical outcome was excellent or good in 80.5% of patients, including 78.3% of patients undergoing a discectomy alone, and 83.3% of patients with posterolateral fusion. The recovery rate was 82.2%, and the difference between the fusion and nonfusion groups was insignificant (P = 0.799). The difference in the postoperative back pain score was also insignificant (P = 0.461). These two groups were not different in terms of age, pain-free interval, and follow-up duration. Intraoperative blood loss, length of surgery, and length of hospitalization were significantly less in patients undergoing discectomy alone than in patients with fusion. CONCLUSIONS: Repeat surgery for recurrent sciatica is effective in cases of true recurrent disc herniation. Disc excision alone is recommended for managing recurrent disc herniation. PMID- 16371914 TI - The learning curve associated with thoracoscopic spinal instrumentation. AB - STUDY DESIGN: Consecutive case prospective radiographic and medical record review. OBJECTIVE: To define the learning curve associated with thoracoscopic spinal instrumentation by evaluating operative data and early outcomes of 1 surgeon's (B.L.) cases. SUMMARY OF BACKGROUND DATA: Thoracoscopic spinal instrumentation for the treatment of thoracic adolescent idiopathic scoliosis has emerged as an alternative to open anterior and posterior techniques. The technique is technically demanding and has been perceived as having a prohibitive learning curve. METHODS: The operative reports, charts, and surgeon's database were used to evaluate operating time, estimated blood loss, levels fused, complication rate, blood transfusions, and curve correction, among other variables. For purposes of analysis, the entire cohort was divided into 2 groups of 28 and 29 patients, respectively, and then 4 groups of 14 patients (the last group with 15) were used for comparison. RESULTS: The records of 57 patients were evaluated. No significant difference in estimated blood loss or number of levels fused was noted for either comparison (P = 0.46 and P = 0.66, respectively). There was no significant difference in blood transfusion requirements, with 7% in group 1 and 18% in group 2 (P = 0.35). Operating time was significantly less after 28 patients were operated on 6.2 +/- 1.3 hours versus 5.3 +/- 1.2 hours (P = 0.011). Percent curve correction was significantly better after 28 cases were performed, 54.4 +/- 17.9 in the former groups versus 65.7 +/- 10.4 in the latter half of cases (P = 0.005). Complications were evenly distributed throughout the series. No significant differences were observed between the 2 groups in terms of rate of complication (P = 0.50). No major complications, such as neurologic deficit or significant hemorrhage, were observed. CONCLUSIONS: The learning curve associated with thoracoscopic spinal instrumentation appears to be acceptable. Significant differences were noted in operating time and percent curve correction after 28 cases. The complication rates remained stable throughout the surgeon's experience. PMID- 16371915 TI - Biomodeling as an aid to spinal instrumentation. AB - STUDY DESIGN: Prospective trial. OBJECTIVE: To develop and validate a new method of spinal stereotaxy. SUMMARY OF BACKGROUND DATA: Biomodeling has been found to be helpful for complex skeletal surgery. Frameless stereotaxy has been used for spinal surgery but has significant limitations. A novel stereotactic technique using biomodels has been developed. METHODS: Twenty patients with complex spinal disorders requiring instrumentation were recruited. A three-dimensional CT scan of their spine was performed, and the data were transferred via a DICOM network to a computer workstation. ANATOMICS BIOBUILD software was used to generate the code required to manufacture exact acrylate biomodels of each spine using rapid prototyping. The biomodels were used to obtain informed consent from patients and to simulate surgery. Simulation was performed using a standard power drill to place trajectory pins into the spinal biomodel. Acrylate drill guides were manufactured using the biomodels and trajectory pins as templates. The biomodels and drill guides were sterilized and used intraoperatively to assist with surgical navigation and the placement of instrumentation. RESULTS: The biomodels were found to be highly accurate and of great assistance in the planning and execution of the surgery. The ability to drill optimum screw trajectories into the biomodel and then accurately replicate the trajectory was judged especially helpful. Accurate screw placement was confirmed with postoperative CT scanning. The design of the first two templates was suboptimal as the contact surface area was too great and complex. Approximately 20 minutes was spent before surgery preparing each biomodel and template. Operating time was reduced, as less reliance on intraoperative radiograph was necessary. Patients stated that the biomodels improved informed consent. CONCLUSIONS: The authors have developed a novel method of spinal stereotaxy using exact plastic copies of the spine manufactured using biomodeling technology. Biomodel spinal stereotaxy is a simple and accurate technique that may have advantages over frameless stereotaxy. PMID- 16371916 TI - Interspinous process decompression system (X STOP) device affords superior outcomes and equal safety to nonoperative therapy. PMID- 16371917 TI - Manipulation of the cervical spine could have induced intracranial hypotension. PMID- 16371918 TI - Registry of randomized controlled trials in transplantation. PMID- 16371922 TI - Low-dose radiation plus rapamycin promotes long-term bone marrow chimerism. AB - BACKGROUND: The ability to achieve significant donor engraftment without fully myeloablative conditioning has revolutionized allogeneic stem cell transplantation. These nonmyeloablative approaches may allow extension of this potentially curative modality to an increasing number of patients including those with non-malignant diseases. Although a number of regimens have been explored, the optimal means of conditioning has not been determined. METHODS: We previously demonstrated that rapamycin (RAPA) has the ability to promote T-cell tolerance even in the presence of costimulation. In the current study, we examine the ability of rapamycin or the calcineurin inhibitor cyclosporine A (CSA) to promote chimerism in a murine haploidentical bone marrow transplantation model. Mice were conditioned with 300 cGy and received either RAPA at 3 mg/kg/day IP, CSA at 20 mg/kg/day IP, or no immunosuppression starting on the day before the transplant and continued for 4 weeks. RESULTS: There was no apparent toxicity, and animals maintained normal blood counts throughout. More importantly, long-term macrochimerism was observed only in the RAPA-treated group. CONCLUSIONS: These results establish a simple, nontoxic, irradiation-based regimen that facilitates engraftment without ablation. This strategy may prove useful in nonmalignant disorders such as hemoglobinopathies in which moderate levels of donor chimerism could prove curative. PMID- 16371921 TI - Inhibition of chronic rejection by antibody induced vascular accommodation in fully allogeneic heart allografts. AB - BACKGROUND: The potential role of altered antibody responses as an effector protective mechanism to induce graft accommodation has been widely investigated in xenogeneic responses. Here we investigate the protective effects of antibody binding to vascular endothelium in a fully mismatched allogeneic model of heart transplantation. METHODS: ACI recipients of WF cardiac grafts were treated either with allochimeric [alpha1h ]-RT1.A class I major histocompatibility complex (MHC) extracts (1 mg/rat, p.v. day 0) or high dose of CsA (10 mg/kg/day, p.o., day 0 6). Cardiac allografts were evaluated at 100 days posttransplant by immunohistology for evidence of chronic rejection and/or vascular accommodation. Activation of apoptotic or antiapoptotic mechanisms was verified by DNA fragmentation (TUNEL) analysis. RESULTS: Allochimeric therapy resulted in inhibition of chronic rejection, absence of neointimal formation and induction of vascular accommodation of fully allogeneic WF hearts in ACI hosts. Such accommodation was evident by IgG and IgM vascular endothelial binding and marked reduction of DNA fragmentation. In contrast, CsA therapy resulted in marked neointimal proliferation, without evidence of vascular accommodation. Immunohistochemical analysis failed to demonstrate vascular endothelial antibody binding. Further, severe chronic rejection following CsA treatment was accompanied by marked DNA fragmentation. CONCLUSION: Alteration of humoral immunity induces vascular accommodation in allogeneic transplantation. Vascular accommodation is the underlying mechanism for inhibition allograft vasculopathy following allochimeric MHC class I therapy. PMID- 16371923 TI - Tacrolimus-induced alopecia in female kidney-pancreas transplant recipients. AB - BACKGROUND: Immunosuppressive drugs given to solid organ transplant recipients may be responsible for cosmetic side effects which can endanger patient compliance. Cyclosporine is associated with hirsutism whereas tacrolimus has been associated with rare cases of alopecia. Since 1998, we have included tacrolimus within the immunosuppressive regimen following kidney-pancreas transplantation. The aim of this study was to evaluate the incidence of alopecia in this population and possible risk factors. METHODS: Between January 1, 1995 and October 31, 2003, 59 consecutive simultaneous kidney-pancreas (SPK) transplants were performed in 58 recipients (27 females and 31 males). The immunosuppressive regimen comprised corticosteroids, calcineurin inhibitor (cyclosporine, n=11; or tacrolimus, n=40) and a purine inhibitor (azathioprine or mycophenolate mofetil). RESULTS: Clinically significant alopecia occurred in 13 patients (28.9%) receiving tacrolimus versus none receiving cyclosporine (P<0.001). Of those who experienced alopecia, 11 were female and two were male (P=0.02). The mean delay between transplantation and alopecia was 422 days (range 100-1,567). Other causes of alopecia were excluded. Treatment of alopecia with topic minoxidil was successful in all cases but one, which required conversion from tacrolimus to cyclosporine. CONCLUSIONS: Alopecia is a frequent complication in women receiving tacrolimus therapy following SPK transplantation. Its pathogenesis is unknown. This cosmetic complication must be discussed with patients before transplantation to minimize the risk of noncompliance. PMID- 16371924 TI - Diagnostic accuracy of the fibrotest in hemodialysis and renal transplant patients with chronic hepatitis C virus. AB - BACKGROUND: An accurate diagnosis of hepatitis C virus (HCV)-related liver lesions is mandatory in dialysis patients and kidney recipients to better define the treatment of and contraindications to kidney transplantation. The aim of this study was to assess the diagnostic accuracy of the fibrotest (a noninvasive method to assess liver fibrosis in HCV on a scale from 0 to 1) in hemodialysis and renal transplant patients infected by chronic HCV. METHODS: In all, 110 patients with biopsy-proven HCV (60 renal transplant recipients and 50 hemodialysis patients), determined using the METAVIR scoring system, were studied. RESULTS: Forty-six percent of patients had fibrosis > or =F2. A positive predictive value of a score >0.6 for the presence of significant fibrosis by comparison with liver biopsy was 71%, and an negative predictive value of < 0.2 for excluding significant fibrosis was 77%, respectively. The areas under the ROC curves for the diagnosis of significant fibrosis were 0.66, 0.47, and 0.71 in the global population, hemodialysis patients, and renal transplant patients, respectively. In all, 75% of patients were correctly classified using the fibrotest. If biopsy was restricted to scores in the intermediate range (< 0.6 and >0.2), the index could reduce the indication for biopsy by 47%. The results did not differ significantly in hemodialysis and renal transplant patients. CONCLUSION: The fibrotest has a diagnostic value in hemodialysis and renal transplant patients which is similar to that reported in the general population (75%) and its use could avoid 32% of liver biopsies if it were interpreted in detail in nephrology patients. PMID- 16371925 TI - Beneficial effects of the bioflavonoids curcumin and quercetin on early function in cadaveric renal transplantation: a randomized placebo controlled trial. AB - BACKGROUND: The bioflavonoids quercetin and curcumin are renoprotective natural antioxidants. We wished to examine their effects on early graft function (EF). METHODS: Between September 2002 and August 2004, 43 dialysis dependent cadaveric kidney recipients were enrolled into a study using Oxy-Q which contains 480 mg of curcumin and 20 mg of quercetin, started after surgery and taken for 1 month. They were randomized into three groups: control (placebo), low dose (one capsule, one placebo) and high dose (two capsules). Delayed graft function (DGF) was defined as first week dialysis need and slow function (SGF) as Cr >2.5 mg/dl by day 10. Category variables were compared by chi squared and continuous variables by Kruskal-Wallis. RESULTS: There were four withdrawals: one by patient choice and three for urine leak. The control group had 2/14 patients with DGF vs. none in either treatment group. Incidence of EF was control 43%, low dose 71% and high dose 93% (P=0.013). Serum creatinine was significantly lower at 2 days (control 7.6+/-2.1, low 5.4+/-0.6, high 3.96+/-.35 P=0.0001) and 30 days (control 1.82+/ .16, low 1.65+/-.09, high 1.33 +/-.1, P=0.03). Acute rejection incidence within 6 months was control 14.3%, low dose 14.3% and high dose 0%. Tremor was detected in 13% of high dose patients vs. 46% of others. Urinary HO-1 was higher in bioflavonoid groups. CONCLUSION: Bioflavonoid therapy improved early graft function. Acute rejection and neurotoxicity were lowest in the high dose group. These bioflavonoids improve early outcomes in cadaveric renal transplantation, possibly through HO-1 induction. PMID- 16371926 TI - Rituximab therapy for de novo mixed cryoglobulinemia in renal transplant patients. AB - BACKGROUND: Type II or III cryoglobulins are fairly prevalent in renal-transplant (RT) patients, and are often related to chronic hepatitis C virus (HCV) infection. However, they rarely result in graft dysfunction. They are sustained by proliferation of oligoclonal B-cells. Systemic B-cell depletion and clinical remission of the systemic effects of cryoglobulins have been achieved in HCV positive immunocompetent patients with a human/mouse chimeric monoclonal antibody that specifically reacts with the CD20 antigen (i.e., rituximab). Thus, this provides the rationale to use rituximab for cryoglobulin-related graft dysfunction in RT patients. METHODS: Three RT patients, of whom one was HCV positive, developed renal-function impairment long after transplantation, as well as de novo nephrotic syndrome (n=2) and severe hypertension (n=2). This latter case was related to type III cryoglobulinemia and was associated with membranoproliferative glomerulonephritis. In addition to their baseline standard immunosuppression, the patients were given weekly rituximab infusions of 375 mg/m (two infusions in patient and four infusions for the other two cases). RESULTS: This treatment resulted in a dramatic improvement in renal parameters, particularly in a sustained remittence of nephrotic syndrome, a sustained clearance of cryoglobulins in two cases, but also in severe infectious complications in two cases. CONCLUSION: We conclude that rituximab therapy is highly effective in cryoglobulin-related renal dysfunction in RT patients; however, due to chronic immunosuppression, this is at the expense of infectious complications. PMID- 16371927 TI - Does uric acid have a pathogenetic role in graft dysfunction and hypertension in renal transplant recipients? AB - BACKGROUND: Uric acid (UA) may play a pathogenetic role in hypertension and kidney disease. We explored the prevalence of hyperuricemia and the relationship of UA to graft function and hypertension in prevalent renal transplant recipients (RTR). METHODS: Baseline and follow-up data were collected on 90 RTR (mean age 51 yrs, 53% male, median transplant duration 7 years). Graft function was estimated using MDRD Study Equation 7. RESULTS: At baseline, 70% RTR had hyperuricemia (UA >7.0 mg/dl (0.42 mmol/L) in men and >6.0 mg/dl (0.36 mmol/L) in women) compared to 80% after 2.2 years (P=0.06). UA was not associated with blood pressure (BP) level but was higher in RTR with a history of hypertension compared to those without (8.6+/-1.8 vs. 7.3+/-2.2 mg/dl, [0.51+/-0.11 vs. 0.43+/-0.13 mmol/L], P=0.003) and in RTR on > or =3 antihypertensive medications compared to those taking less (9.1+/-1.6 vs. 7.6+/-1.8 mg/dL, [0.54+/-0.1 vs. 0.45+/-0.11 mmol/L], P<0.001). A history of hypertension was independently predictive of UA (beta 0.06, [95% CI 0.02 to 0.10], P=0.007) in addition to sex, cyclosporine dose, prednisolone dose, estimated glomerular filtration rate (eGFRMDRD) and beta blocker therapy. UA was independently predictive of follow-up eGFRMDRD (beta 22.2 [95% CI -41.2 to -3.2], P=0.02) but did not predict change in eGFRMDRD over time. UA was independently associated with requirement for antihypertensive therapy (beta 0.34, [95% CI 1.05 to 1.90], P=0.02). CONCLUSIONS: Hyperuricemia is common in RTR and is associated with need for antihypertensive therapy and level of graft function. PMID- 16371928 TI - A Comparison of splenectomy versus intensive posttransplant antidonor blood group antibody monitoring without splenectomy in ABO-incompatible kidney transplantation. AB - BACKGROUND: Although most protocols for ABO-incompatible kidney transplantation have employed splenectomy, its utility is unproven. The aim of the current study was to compare the outcomes of ABO-incompatible living donor kidney transplantation with splenectomy versus a protocol involving intensive posttransplant antibody monitoring to maintain low levels of antiblood group antibody. METHODS: We retrospectively studied all ABO-incompatible living donor kidney transplants at our institution between September 1999 and November 2004 (n=34). Prior to May 2003, all patients were included in a protocol involving pretransplant plasmapheresis and splenectomy at the time of transplant (n=23). After May 2003, splenectomy was not performed and a protocol that involved pretransplant anti-CD20 antibody and a more intensive posttransplant plasmapheresis regiment aimed at maintaining low levels of antiblood group antibody during the first 2 weeks following transplantation was utilized (n=11). RESULTS: Patient and graft survival was similar in the two groups. Humoral rejection occurred in 18% nonsplenectomized and 30% of splenectomized patients (P=0.68). Humoral rejection correlated with the baseline antibody titer in both groups. Individuals with elevated baseline antibody titer (> or =1:256) appear to be at high risk for humoral rejection regardless of protocol used. Antiblood group antibody levels 3 and 12 months after transplantation were similar in both groups. CONCLUSIONS: Splenectomy is not essential for successful ABO-incompatible kidney transplantation, although individuals with high baseline antidonor blood group antibody titers are at high risk for humoral rejection. The use of intensive posttransplant monitoring may help prevent antibody-mediated graft damage. PMID- 16371929 TI - Conversion from cyclosporine to sirolimus in stable renal transplant recipients. AB - BACKGROUND: Conversion from cyclosporine (CsA) to sirolimus (SRL) has mainly been done in clinical conditions warranting calcineurin inhibitor discontinuation. Little is known about the clinical outcome of conversion in renal transplant recipients without transplant dysfunction. METHODS: This prospective, open-label, multicentric pilot study evaluates the safety and efficacy of converting patients with stable renal function from CsA to SRL. RESULTS: Forty stable patients on CsA, mycophenolate mofetil (MMF) (1.5 g/day), and steroids (ST) were converted at 7.6+/-1.4 months after renal transplantation. At 1 year, graft and patient survival was 100% and the incidence of acute rejection 5%. Calculated glomerular filtration rate (GFR) increased from 54+/-18 to 66+/-16 ml/min (P<0.0001). Blood pressure remained unchanged. A gradual increase in the incidence and severity of proteinuria was observed from month 6 onwards with de novo proteinuria in 30% of the patients at 1 year. Protein excretion was below 1 g/day in 12.5%, between 1 and 3 g/day in 17.5% and above 3 g/day in 7.5% of the proteinuric cohort (P=0.0043, compared to baseline). No predictors could be identified for the development of proteinuria. All patients had a reduction in protein excretion following renin-angiotensin blockade and were continued on SRL. CONCLUSION: Conversion of stable renal transplant recipients from a CsA-MMF-ST to a SRL-MMF ST regimen is safe and results in improved renal function but is associated with the development of proteinuria in 30% of the patients requiring renin-angiotensin blockade. PMID- 16371930 TI - Tissue cardiomyoplasty using bioengineered contractile cardiomyocyte sheets to repair damaged myocardium: their integration with recipient myocardium. AB - BACKGROUND: We hypothesized that tissue-engineered contractile cardiomyocyte sheets without a scaffold would show histological and electrical integration with impaired myocardium, leading to the regeneration of infarcted myocardium. METHODS: Neonatal rat cardiomyocytes were cultured on Poly(N-isopropylacrylamide) grafted polystyrene dishes and detached as a square cell sheet at 20 degrees C. Two sheets were stacked to make thicker contractile cardiac sheets. In cross section, the stacked sheets looked like homogeneous heart-like tissue. Two weeks after rats were subjected to left anterior descending (LAD) ligation, two treatments were conducted: 1) cardiomyocyte sheet implantation (T group, n=10), and 2) fibroblast sheet implantation (F group, n=10). The control group underwent no additional treatment (C group, n=10). RESULTS: Echocardiography demonstrated that cardiac performance was significantly ameliorated in the T group 2, 4, and 8 weeks after implantation. The cardiomyocyte sheets became attached to the infarcted myocardium, showed angiogenesis, expressed connexin-43, and appeared as homogeneous tissue in the myocardium Electrophysiological experiments showed a QRS complex with one peak in the treated scar area in the T group, but two peaks, indicative of branch block, in that of the other groups. Furthermore, the threshold for pacing of the recipient heart was lower in the T group than in the other groups. CONCLUSIONS: Cardiomyocyte sheets integrated with the impaired myocardium and improved cardiac performance in a model of ischemic myocardium. Techniques using such tissue-engineered cell sheets are introducing the promising concept of tissue cardiomyoplasty to the field of regenerative medicine. PMID- 16371931 TI - Magnetic resonance imaging of pancreatic islets in tolerance and rejection. AB - BACKGROUND: We have recently described a magnetic resonance (MR) method for detection of rat pancreatic islets transplanted into the liver after labeling with superparamagnetic iron oxide nanoparticles. The aim of this work was to study whether this technique could be applicable over a longer period after transplantation and whether it could help to detect islet rejection. METHODS: Islets from Lewis and Wistar rats were cultured in the presence of iron oxide nanoparticles. Two thousand of Lewis (n=6) or Wistar (n=8) iron-labeled islets were transplanted into the portal vein of Lewis diabetic animals. Serial MR imaging of the liver were performed at 1, 2, 3, 4, 5, and 6 weeks. RESULTS: Although all allogeneic islets were rejected by 12 days, syngeneic animals remained normoglycemic throughout the study. At week 1, the labeled islets were visualized on MR scans as distinct hypointense spots homogeneously distributed in the liver. While their number declined only insignificantly in the syngeneic group, in the allogeneic group the number of spots gradually decreased until approximately 35% of their initial count. Although syngeneic islets showed a normal histology, the allogeneic islets were completely rejected. Iron particles, localized in macrophages, were detected only in the syngeneic islets and were absent in the rejected islet structures. In vitro incubation tests did not reveal any differences in insulin secretion between labeled and nonlabeled islets. CONCLUSIONS: MR imaging of iron-labeled pancreatic islets can be used for verification of the technical success of the transplantation procedure itself and for the detection of the decreasing relative islet mass due to rejection. PMID- 16371932 TI - Sex steroid ablation enhances lymphoid recovery following autologous hematopoietic stem cell transplantation. AB - BACKGROUND: Autologous hematopoietic stem cell transplantation (auto-HSCT) patients experience long-term immunosuppression, which increases susceptibility to infection and relapse rates due to minimal residual disease (MRD). Sex steroid (SS) ablation is known to reverse age-related thymic atrophy and decline in B cell production METHODS: This study used a congenic HSCT mouse model to analyze the effects of SS ablation (through surgical castration) on immune reconstitution and growth factor production following auto-HSCT. Bone marrow (BM) and thymic stromal cell (TSCs) populations were analyzed using RT-PCR and were tested for the production of growth factors previously implicated in immune reconstitution or age-relate immune degeneration RESULTS: Castration increased bone marrow (BM), thymic, and splenic cellularity following auto-HSCT. HSC number and common lymphoid precursor (CLP) frequency and number were increased in castrated mice. B cell precursor numbers were also significantly increased in the BM of these mice. Triple negative, double positive and single positive thymocytes were increased following HSCT and castration, as were thymic dendritic cells and natural killer T (NKT) cells. This enhanced lymphoid reconstitution of the primary immune organs leads to a significant increase in splenic T and B cells 42 days after HSCT. The molecular mechanisms behind the enhanced reconstitution were also studied. TGF beta1 was decreased in castrated mice compared to sham-castrated controls in TSCs and BM cells. TSC production of IL-6 was also decreased in castrated mice CONCLUSIONS: These data suggest that sex steroid ablation significantly enhances lymphopoiesis following auto-HSCT providing a new strategy for posttransplant immune reconstitution. PMID- 16371933 TI - Dimeric but not monomeric soluble CD40 prolongs allograft survival and generates regulatory T cells that inhibit CTL function. AB - BACKGROUND: We previously reported that adenovirus mediated CD40Ig gene therapy (AdCD40Ig) induced long-term acceptance of fully allogeneic rat cardiac allografts, however, the underlying mechanism has not been fully clarified. To address this we have compared the ability of dimeric and monomeric soluble CD40 to prolong allograft survival in vivo and generate regulatory T cells in vitro. METHODS: The ability of CD40Ig (soluble dimmer, containing an Fc region) or CD40/Myc/His (soluble monomer, lacking an Fc region) therapy to generate CD4CD25 regulatory T cells in vitro and to prevent rejection of rat cardiac allografts (ACI to LEWIS) was compared. Immunoregulatory capacity of regulatory T cells generated was determined by suppression of alloantigen specific proliferation and cytotoxicity. RESULTS: Dimeric soluble CD40Ig did not inhibit CD4 T cell proliferation but rather promoted IL-2 production and the generation of CD4CD25 T cells, which regulated alloantigen-specific cytotoxic T lymphocyte activity. Treatment with either AdCD40Ig or purified soluble CD40Ig prolonged the survival of rat cardiac allografts. In contrast, although monomeric soluble CD40/Myc/His suppressed IL-12 production in a similar manner to that achieved by CD40Ig, it did not augment IL-2 production. Moreover, while CD40/Myc/His also generated CD4CD25 T cells, they did not exhibit regulatory activity and administration of soluble CD40/Myc/His failed to prolong cardiac allograft survival. CONCLUSIONS: These results suggest signaling through CD154 in addition to blocking of CD154 CD40 interaction is important for the immunomodulatory effects of soluble CD40Ig. Taken together, our results provide new insight into the mechanism of immunomodulation by soluble CD40 constructs. PMID- 16371934 TI - The efficacy and safety of gene transfer into the porcine liver in vivo by HVJ (Sendai virus) liposome. AB - BACKGROUND: Gene transfer systems using viral vectors are efficient; however, most viral vectors also tend to evoke immunologic reactions, thereby clinically causing serial side effects. HVJ-liposome vector is a hybrid vector consisting of liposome and an inactivated Sendai virus (Hemmagglutinating Virus of Japan [HVJ]), which has been reported to be less immunogenic and can also be repeatedly administered. We examined the usefulness of this vector for hepatic gene therapy in a pig model. METHODS: Genes encoding beta-galactosidase and luciferase were used as reporter genes. The pigs were injected with the reporter gene loaded-HVJ liposome into the portal vein under total vascular exclusion of the liver. The transfection efficiencies were then assessed by beta-galactosidase staining, a luciferase assay, and RT-PCR for LacZ mRNA. Biochemical and histologic analyses were performed to evaluate tissue toxicity after gene transfer. RESULTS: The luciferase gene expression in the liver reached its highest level at 7 days after transfection. It continued to be detected up to 28 days after transfection, while all pigs remained healthy throughout the observation period. The transfection efficiency was 15% in the hepatocytes according to beta-galactosidase staining. Extrahepatic transgene expression was slightly observed in the lung and kidney, but not in the spleen or ovary. CONCLUSIONS: These data suggest for the first time that the use of the HVJ-liposome vector is a safe and feasible modality for liver-directed gene transfer in pigs, and it might therefore be suitable for clinical gene therapy trials. PMID- 16371935 TI - Liver transplantation in sickle cell anemia: a case of acute sickle cell intrahepatic cholestasis and a case of sclerosing cholangitis. AB - Very few cases of liver transplantation in patients with sickle cell disease have been reported in peer-reviewed literature. We reviewed the medical records of two patients with sickle cell disease that received liver transplantation at our institution. The first patient was a 27-year-old female who presented with encephalopathy and cholestatic jaundice with a Hemoglobin S (HbS) level of 69.6%. She was diagnosed with acute sickle cell intrahepatic cholestasis. The second patient was a 26-year-old female with sclerosing cholangitis who presented with encephalopathy, bleeding, and cholestatic jaundice. Her HbS level was normal. Both patients underwent liver transplantation successfully but died in the postoperative period from multiorgan failure. We report a rare case of liver transplantation for acute sickle cell intrahepatic cholestasis and a novel case of transplantation in a patient with sickle cell disease and sclerosing cholangitis. Liver transplantation did not lead to a successful outcome in either case. PMID- 16371936 TI - Does bioequivalence between modified cyclosporine formulations translate into equal outcomes? AB - Neoral was replaced with a generic cyclosporine formulation on our hospital formulary. We compared outcomes for de novo kidney transplant recipients who either received Gengraf (n=88) or Neoral (n=100) in a single-center, retrospective review. As compared to patients who received Neoral, patients who received Gengraf were significantly more likely to have an acute rejection episode (39% vs. 25%, P=0.04), more likely to have a second rejection episode (13% vs. 4%; P=0.03), or to have received an antibody preparation to treat acute rejection (19% vs. 8%; P=0.02). Patients treated with Gengraf had a higher degree of intrapatient variability for cyclosporine trough concentrations as determined by %CV (P<0.05). The incidence of acute rejection at 6 months posttransplant was significantly higher in patients who received Gengraf compared to Neoral. A larger, prospective analysis is warranted to compare these formulations of cyclosporine in de novo kidney transplant recipients. PMID- 16371937 TI - The cost effectiveness of deceased organ donation. PMID- 16371938 TI - Ex vivo tumorectomy on transplanted kidney. PMID- 16371939 TI - Significant coronary artery disease appears to have increased in the past 21 years in diabetics being evaluated for organ transplantation. PMID- 16371940 TI - Pretransplant workup in renal transplantation; soluble CD30 versus panel reactive antibody. PMID- 16371941 TI - Personal and organizational preparedness. The key to surviving a disaster. PMID- 16371942 TI - Outbreaks of Escherichia coli O157:H7 associated with petting zoos--North Carolina, Florida, and Arizona, 2004 and 2005. AB - During 2004-2005, three outbreaks of Escherichia coli O157:H7 infections occurred among agricultural fair, festival, and petting zoo visitors in North Carolina, Florida, and Arizona. One hundred eight cases, including 15 cases of hemolytic uremic syndrome (HUS), were reported in the North Carolina outbreak; 63 cases, including seven HUS cases, were reported in the Florida outbreak; and two cases were reported in Arizona. No fatalities occurred. Illnesses primarily affected children who visited petting zoos at these events. This report summarizes findings from these outbreak investigations, which indicated the need for adequate control measures to reduce zoonotic transmission of E. coli O157:H7. PMID- 16371943 TI - Mycobacterium tuberculosis transmission in a newborn nursery and maternity ward- New York City, 2003. AB - Evaluating young children recently exposed to airborne Mycobacterium tuberculosis is a public health priority. If infected, children aged <2 years are at high risk for severe tuberculosis (TB) disease (e.g., TB meningitis). In December 2003, infectious pulmonary TB disease was diagnosed in a foreign-born nurse working in the newborn nursery and maternity ward of a New York City hospital (hospital A); the nurse had declined treatment for latent TB infection (LTBI) after testing positive 11 years earlier. An investigation including medical evaluation of contacts in the nursery and maternity ward was conducted by the Bureau of TB Control (BTBC) at the New York City Department of Health and Mental Hygiene, hospital A, and CDC. This report summarizes the results of that investigation, which determined that approximately 1,500 patients had been exposed to the nurse but the majority could not be located for evaluation. Among those who were tested, four infants had positive tuberculin skin test (TST) results, likely attributable to recent transmission of M. tuberculosis. The findings emphasize the difficulty of conducting contact investigations in certain settings and the importance of effective LTBI testing and treatment programs for health-care workers (HCWs) to prevent TB disease and subsequent health-care--associated transmission. PMID- 16371944 TI - Pertussis--United States, 2001-2003. AB - Pertussis is a highly contagious, vaccine-preventable bacterial illness characterized by paroxysmal cough, posttussive vomiting, and inspiratory whoop. Pertussis also can occur as a mild or moderate cough illness in persons who are partially immune. In the United States, most hospitalizations and nearly all deaths from pertussis are reported in infants aged <6 months, but substantial morbidity does occur in other age groups. Infant/childhood vaccination has contributed to a reduction of more than 90% in pertussis-related morbidity and mortality since the early 1940s in the United States. Estimates of childhood vaccination coverage with > or =3 doses of pertussis-containing vaccine have exceeded 90% since 1994; however, reported pertussis cases increased from a historic low of 1,010 in 1976 to 11,647 cases in 2003. A substantial increase in reported cases has occurred among adolescents, who become susceptible to pertussis approximately 6-10 years after childhood vaccination. Recently, booster vaccines for adolescents and adults combining pertussis antigens with tetanus and diphtheria toxoids (Tdap) were approved by the Food and Drug Administration (FDA). On June 30, 2005, the Advisory Committee on Immunization Practices (ACIP) recommended Tdap for all persons aged 11-18 years. This report summarizes national surveillance data on pertussis reported to CDC during 2001-2003 and focuses on pertussis reported among persons aged 10-19 years before implementation of national recommendations for adolescent pertussis vaccination. PMID- 16371945 TI - A comprehensive immunization strategy to eliminate transmission of hepatitis B virus infection in the United States: recommendations of the Advisory Committee on Immunization Practices (ACIP) part 1: immunization of infants, children, and adolescents. AB - This report is the first of a two-part statement from the Advisory Committee on Immunization Practices (ACIP) that updates the strategy to eliminate hepatitis B virus (HBV) transmission in the United States. The report provides updated recommendations to improve prevention of perinatal and early childhood HBV transmission, including implementation of universal infant vaccination beginning at birth, and to increase vaccine coverage among previously unvaccinated children and adolescents. Strategies to enhance implementation of the recommendations include 1) establishing standing orders for administration of hepatitis B vaccination beginning at birth; 2) instituting delivery hospital policies and procedures and case management programs to improve identification of and administration of immunoprophylaxis to infants born to mothers who are hepatitis B surface antigen (HBsAg) positive and to mothers with unknown HBsAg status at the time of delivery; and 3) implementing vaccination record reviews for all children aged 11-12 years and children and adolescents aged <19 years who were born in countries with intermediate and high levels of HBV endemicity, adopting hepatitis B vaccine requirements for school entry, and integrating hepatitis B vaccination services into settings that serve adolescents. The second part of the ACIP statement, which will include updated recommendations and strategies to increase hepatitis B vaccination of adults, will be published separately. PMID- 16371946 TI - Oligodendrocyte wars. AB - Oligodendrocyte precursors first arise in a restricted ventral part of the embryonic spinal cord and migrate laterally and dorsally from there. Later, secondary sources develop in the dorsal cord. Normally, the ventrally-derived precursors compete with and suppress their dorsal counterparts. There are also ventral and dorsal sources in the forebrain, but here the more dorsal precursors prevail and the ventral-most lineage is eliminated during postnatal life. How do the different populations compete and what is the outcome of the competition? Do different embryonic origins signify different functional subgroups of oligodendrocyte? PMID- 16371947 TI - The sensory and motor roles of auditory hair cells. AB - Cochlear hair cells respond with phenomenal speed and sensitivity to sound vibrations that cause submicron deflections of their hair bundle. Outer hair cells are not only detectors, but also generate force to augment auditory sensitivity and frequency selectivity. Two mechanisms of force production have been proposed: contractions of the cell body or active motion of the hair bundle. Here, we describe recently identified proteins involved in the sensory and motor functions of auditory hair cells and present evidence for each force generator. Both motor mechanisms are probably needed to provide the high sensitivity and frequency discrimination of the mammalian cochlea. PMID- 16371948 TI - Neural plasticity in the ageing brain. AB - The mechanisms involved in plasticity in the nervous system are thought to support cognition, and some of these processes are affected during normal ageing. Notably, cognitive functions that rely on the medial temporal lobe and prefrontal cortex, such as learning, memory and executive function, show considerable age related decline. It is therefore not surprising that several neural mechanisms in these brain areas also seem to be particularly vulnerable during the ageing process. In this review, we discuss major advances in our understanding of age related changes in the medial temporal lobe and prefrontal cortex and how these changes in functional plasticity contribute to behavioural impairments in the absence of significant pathology. PMID- 16371949 TI - Astrocyte-endothelial interactions at the blood-brain barrier. AB - The blood-brain barrier, which is formed by the endothelial cells that line cerebral microvessels, has an important role in maintaining a precisely regulated microenvironment for reliable neuronal signalling. At present, there is great interest in the association of brain microvessels, astrocytes and neurons to form functional 'neurovascular units', and recent studies have highlighted the importance of brain endothelial cells in this modular organization. Here, we explore specific interactions between the brain endothelium, astrocytes and neurons that may regulate blood-brain barrier function. An understanding of how these interactions are disturbed in pathological conditions could lead to the development of new protective and restorative therapies. PMID- 16371950 TI - Cognitive neuroscience of emotional memory. AB - Emotional events often attain a privileged status in memory. Cognitive neuroscientists have begun to elucidate the psychological and neural mechanisms underlying emotional retention advantages in the human brain. The amygdala is a brain structure that directly mediates aspects of emotional learning and facilitates memory operations in other regions, including the hippocampus and prefrontal cortex. Emotion-memory interactions occur at various stages of information processing, from the initial encoding and consolidation of memory traces to their long-term retrieval. Recent advances are revealing new insights into the reactivation of latent emotional associations and the recollection of personal episodes from the remote past. PMID- 16371951 TI - Neuroscience nanotechnology: progress, opportunities and challenges. AB - Nanotechnologies exploit materials and devices with a functional organization that has been engineered at the nanometre scale. The application of nanotechnology in cell biology and physiology enables targeted interactions at a fundamental molecular level. In neuroscience, this entails specific interactions with neurons and glial cells. Examples of current research include technologies that are designed to better interact with neural cells, advanced molecular imaging technologies, materials and hybrid molecules used in neural regeneration, neuroprotection, and targeted delivery of drugs and small molecules across the blood-brain barrier. PMID- 16371952 TI - Gene therapy: can neural stem cells deliver? AB - Neural stem cells are a self-renewing population that generates the neurons and glia of the developing brain. They can be isolated, proliferated, genetically manipulated and differentiated in vitro and reintroduced into a developing, adult or pathologically altered CNS. Neural stem cells have been considered for use in cell replacement therapies in various neurodegenerative diseases, and an unexpected and potentially valuable characteristic of these cells has recently been revealed--they are highly migratory and seem to be attracted to areas of brain pathology such as ischaemic and neoplastic lesions. Here, we speculate on the ways in which neural stem cells might be exploited as delivery vehicles for gene therapy in the CNS. PMID- 16371953 TI - Meteorology: hurricanes and global warming. AB - Anthropogenic climate change has the potential for slightly increasing the intensity of tropical cyclones through warming of sea surface temperatures. Emanuel has shown a striking and surprising association between sea surface temperatures and destructiveness by tropical cyclones in the Atlantic and western North Pacific basins. However, I question his analysis on the following grounds: it does not properly represent the observations described; the use of his Atlantic bias-removal scheme may not be warranted; and further investigation of a substantially longer time series for tropical cyclones affecting the continental United States does not show a tendency for increasing destructiveness. These factors indicate that instead of "unprecedented" tropical cyclone activity having occurred in recent years, hurricane intensity was equal or even greater during the last active period in the mid-twentieth century. PMID- 16371954 TI - Meteorology: are there trends in hurricane destruction? AB - Since the record impact of Hurricane Katrina, attention has focused on understanding trends in hurricanes and their destructive potential. Emanuel reports a marked increase in the potential destructiveness of hurricanes based on identification of a trend in an accumulated annual index of power dissipation in the North Atlantic and western North Pacific since the 1970s. If hurricanes are indeed becoming more destructive over time, then this trend should manifest itself in more destruction. However, my analysis of a long-term data set of hurricane losses in the United States shows no upward trend once the data are normalized to remove the effects of societal changes. PMID- 16371959 TI - A poor assessment. PMID- 16371960 TI - AIDS at Christmas time. PMID- 16371961 TI - A recipe for trouble. PMID- 16371963 TI - Korean scandal will have global fallout. PMID- 16371964 TI - Where now for stem-cell cloners? PMID- 16371965 TI - Dogged by doubts. PMID- 16371966 TI - Diet book attacked for its high-protein advice. PMID- 16371967 TI - India makes waves over tsunami warning system. PMID- 16371969 TI - The heat was on in 2005. PMID- 16371971 TI - First glimpse... PMID- 16371972 TI - Alcohol and science: the party gene. PMID- 16371973 TI - Alcohol and science: saving the agave. PMID- 16371974 TI - Alcohol and science: the grapes of rock. PMID- 16371975 TI - Merck opts for shake-up to clear drug pipeline. PMID- 16371978 TI - Network aims to make maths count in Africa. PMID- 16371979 TI - Women's efforts are more than a drop in the ocean. PMID- 16371980 TI - Animal culture is real but needs to be clearly defined. PMID- 16371981 TI - For quiet students, finding a voice is the first step towards taking a stand. PMID- 16371982 TI - Barriers to progress in systems biology. PMID- 16371987 TI - Unravelling string theory. PMID- 16371988 TI - The death of a star. PMID- 16371989 TI - Mars: the flow and ebb of water. PMID- 16371990 TI - Physics: philately will get you everywhere. PMID- 16371991 TI - Molecular biology: antagonizing the neighbours. PMID- 16371992 TI - Atmospheric physics: reflections on aerosol cooling. PMID- 16371993 TI - Genomics: multiple moulds. PMID- 16371995 TI - Obituary: Richard E. Smalley (1943-2005). PMID- 16371996 TI - Circadian organization in reindeer. AB - The light/dark cycle of day and night synchronizes an internal 'biological clock' that governs daily rhythms in behaviour, but this form of regulation is denied to polar animals for most of the year. Here we demonstrate that the continuous lighting conditions of summer and of winter at high latitudes cause a loss in daily rhythmic activity in reindeer living far above the Arctic Circle. This seasonal absence of circadian rhythmicity may be a ubiquitous trait among resident polar vertebrates. PMID- 16371997 TI - World Year of Physics: a direct test of E=mc2. AB - One of the most striking predictions of Einstein's special theory of relativity is also perhaps the best known formula in all of science: E=mc(2). If this equation were found to be even slightly incorrect, the impact would be enormous- given the degree to which special relativity is woven into the theoretical fabric of modern physics and into everyday applications such as global positioning systems. Here we test this mass-energy relationship directly by combining very accurate measurements of atomic-mass difference, Delta(m), and of gamma-ray wavelengths to determine E, the nuclear binding energy, for isotopes of silicon and sulphur. Einstein's relationship is separately confirmed in two tests, which yield a combined result of 1-Delta(mc2)/E=(-1.4+/-4.4)x10(-7), indicating that it holds to a level of at least 0.00004%. To our knowledge, this is the most precise direct test of the famous equation yet described. PMID- 16371998 TI - Chemical communication: chirality in elephant pheromones. AB - Musth in male elephants is an annual period of heightened sexual activity and aggression that is linked to physical, sexual and social maturation. It is mediated by the release of chemical signals such as the pheromone frontalin, which exists in two chiral forms (molecular mirror images, or enantiomers). Here we show that enantiomers of frontalin are released by Asian elephants in a specific ratio that depends on the animal's age and stage of musth, and that different responses are elicited in male and female conspecifics when the ratio alters. This precise control of communication by molecular chirality offers insight into societal interactions in elephants, and may be useful in implementing new conservation protocols. PMID- 16371999 TI - An Asian perspective on early human dispersal from Africa. AB - The past decade has seen the Pliocene and Pleistocene fossil hominin record enriched by the addition of at least ten new taxa, including the Early Pleistocene, small-brained hominins from Dmanisi, Georgia, and the diminutive Late Pleistocene Homo floresiensis from Flores, Indonesia. At the same time, Asia's earliest hominin presence has been extended up to 1.8 Myr ago, hundreds of thousands of years earlier than previously envisaged. Nevertheless, the preferred explanation for the first appearance of hominins outside Africa has remained virtually unchanged. We show here that it is time to develop alternatives to one of palaeoanthropology's most basic paradigms: 'Out of Africa 1'. PMID- 16372000 TI - Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae. AB - The aspergilli comprise a diverse group of filamentous fungi spanning over 200 million years of evolution. Here we report the genome sequence of the model organism Aspergillus nidulans, and a comparative study with Aspergillus fumigatus, a serious human pathogen, and Aspergillus oryzae, used in the production of sake, miso and soy sauce. Our analysis of genome structure provided a quantitative evaluation of forces driving long-term eukaryotic genome evolution. It also led to an experimentally validated model of mating-type locus evolution, suggesting the potential for sexual reproduction in A. fumigatus and A. oryzae. Our analysis of sequence conservation revealed over 5,000 non-coding regions actively conserved across all three species. Within these regions, we identified potential functional elements including a previously uncharacterized TPP riboswitch and motifs suggesting regulation in filamentous fungi by Puf family genes. We further obtained comparative and experimental evidence indicating widespread translational regulation by upstream open reading frames. These results enhance our understanding of these widely studied fungi as well as provide new insight into eukaryotic genome evolution and gene regulation. PMID- 16372001 TI - Impact origin of sediments at the Opportunity landing site on Mars. AB - Mars Exploration Rover Opportunity discovered sediments with layered structures thought to be unique to aqueous deposition and with minerals attributed to evaporation of an acidic salty sea. Remarkable iron-rich spherules were ascribed to later groundwater alteration, and the inferred abundance of water reinforced optimism that Mars was once habitable. The layered structures, however, are not unique to water deposition, and the scenario encounters difficulties in accounting for highly soluble salts admixed with less soluble salts, the lack of clay minerals from acid-rock reactions, high sphericity and near-uniform sizes of the spherules and the absence of a basin boundary. Here we present a simple alternative explanation involving deposition from a ground-hugging turbulent flow of rock fragments, salts, sulphides, brines and ice produced by meteorite impact. Subsequent weathering by intergranular water films can account for all of the features observed without invoking shallow seas, lakes or near-surface aquifers. Layered sequences observed elsewhere on heavily cratered Mars and attributed to wind, water or volcanism may well have formed similarly. If so, the search for past life on Mars should be reassessed accordingly. PMID- 16372002 TI - A volcanic environment for bedrock diagenesis at Meridiani Planum on Mars. AB - Exposed bedrocks at Meridiani Planum on Mars display chemical and mineralogical evidence suggesting interaction with liquid water. On the basis of morphological observations as well as high abundances of haematite and sulphate minerals, the rocks have been interpreted as sediments that were deposited in a shallow body of briny water with subsequent evaporation leaving behind the sulphate minerals. The iron-sulphur mineralization at Meridiani has also been inferred to be analogous to that produced during oxidative weathering of metal sulphide minerals, such as occurs at acid mine drainage sites. Neither of these interpretations, however, is consistent with the chemical composition of the rocks. Here we propose an alternative model for diagenesis of Meridiani bedrock that involves deposition of volcanic ash followed by reaction with condensed sulphur dioxide- and water bearing vapours emitted from fumaroles. This scenario does not require prolonged interaction with a standing body of surface water and may have occurred at high temperatures. Consequently, the model invokes an environment considerably less favourable for biological activity on Mars than previously proposed interpretations. PMID- 16372003 TI - Light echoes from ancient supernovae in the Large Magellanic Cloud. AB - The light from historical supernovae could in principle still be visible as scattered-light echoes centuries after the explosion. The detection of light echoes could allow us to pinpoint the supernova event both in position and age and, most importantly, permit the acquisition of spectra to determine the 'type' of the supernova centuries after the direct light from the explosion first reached Earth. Although echoes have been discovered around some nearby extragalactic supernovae, targeted searches have not found any echoes in the regions of historical Galactic supernovae. Here we report three faint variable surface-brightness complexes with high apparent proper motions pointing back to three of the six smallest (and probably youngest) previously catalogued supernova remnants in the Large Magellanic Cloud, which are believed to have been thermonuclear (type Ia) supernovae. Using the distance and apparent proper motions of these echo arcs, we estimate ages of 610 and 410 years for two of them. PMID- 16372004 TI - Extremely slow Drude relaxation of correlated electrons. AB - The electrical conduction of metals is governed by how freely mobile electrons can move throughout the material. This movement is hampered by scattering with other electrons, as well as with impurities or thermal excitations (phonons). Experimentally, the scattering processes of single electrons are not observed, but rather the overall response of all mobile charge carriers within a sample. The ensemble dynamics can be described by the relaxation rates, which express how fast the system approaches equilibrium after an external perturbation. Here we measure the frequency-dependent microwave conductivity of the heavy-fermion metal UPd2Al3 (ref. 4), finding that it is accurately described by the prediction for a single relaxation rate (the so-called Drude response). This is notable, as UPd2Al3 has strong interactions among the electrons that might be expected to lead to more complex behaviour. Furthermore, the relaxation rate of just a few gigahertz is extremely low--this is several orders of magnitude below those of conventional metals (which are typically around 10 THz), and at least one order of magnitude lower than previous estimates for comparable metals. These observations are directly related to the high effective mass of the charge carriers in this material and reveal the dynamics of interacting electrons. PMID- 16372005 TI - Global estimate of aerosol direct radiative forcing from satellite measurements. AB - Atmospheric aerosols cause scattering and absorption of incoming solar radiation. Additional anthropogenic aerosols released into the atmosphere thus exert a direct radiative forcing on the climate system. The degree of present-day aerosol forcing is estimated from global models that incorporate a representation of the aerosol cycles. Although the models are compared and validated against observations, these estimates remain uncertain. Previous satellite measurements of the direct effect of aerosols contained limited information about aerosol type, and were confined to oceans only. Here we use state-of-the-art satellite based measurements of aerosols and surface wind speed to estimate the clear-sky direct radiative forcing for 2002, incorporating measurements over land and ocean. We use a Monte Carlo approach to account for uncertainties in aerosol measurements and in the algorithm used. Probability density functions obtained for the direct radiative forcing at the top of the atmosphere give a clear-sky, global, annual average of -1.9 W m(-2) with standard deviation, +/- 0.3 W m(-2). These results suggest that present-day direct radiative forcing is stronger than present model estimates, implying future atmospheric warming greater than is presently predicted, as aerosol emissions continue to decline. PMID- 16372006 TI - Anisotropy of Earth's D'' layer and stacking faults in the MgSiO3 post-perovskite phase. AB - The post-perovskite phase of (Mg,Fe)SiO3 is believed to be the main mineral phase of the Earth's lowermost mantle (the D'' layer). Its properties explain numerous geophysical observations associated with this layer-for example, the D'' discontinuity, its topography and seismic anisotropy within the layer. Here we use a novel simulation technique, first-principles metadynamics, to identify a family of low-energy polytypic stacking-fault structures intermediate between the perovskite and post-perovskite phases. Metadynamics trajectories identify plane sliding involving the formation of stacking faults as the most favourable pathway for the phase transition, and as a likely mechanism for plastic deformation of perovskite and post-perovskite. In particular, the predicted slip planes are {010} for perovskite (consistent with experiment) and {110} for post-perovskite (in contrast to the previously expected {010} slip planes). Dominant slip planes define the lattice preferred orientation and elastic anisotropy of the texture. The {110} slip planes in post-perovskite require a much smaller degree of lattice preferred orientation to explain geophysical observations of shear-wave anisotropy in the D'' layer. PMID- 16372007 TI - The pelvic fin and girdle of Panderichthys and the origin of tetrapod locomotion. AB - One of the most marked transformations in the vertebrate transition to land was that of fins to limbs. This transformation involved not only the generation of morphological novelties (digits, sacrum) but also a shift in locomotory dominance from the pectoral to the pelvic appendage. Despite its importance, the transformation from pelvic fin to hindlimb is the least studied and least well documented part of this transformation, which is bracketed by the osteolepiform Eusthenopteron and the early tetrapods Ichthyostega and Acanthostega, but is not directly illuminated by any intermediate form. Panderichthys is the closest tetrapod relative currently represented by complete fossils, but its pelvic fin skeleton has not been described. Here, I present the only known articulated pelvic fin endoskeleton and associated partial pelvis of Panderichthys. The pelvic girdle is even less tetrapod-like than that of the osteolepiform Eusthenopteron, but the pelvic fin endoskeleton shares derived characteristics with basal tetrapods despite being more primitive than the pectoral fin of Panderichthys. The evolution of tetrapod locomotion appears to have passed through a stage of body-flexion propulsion, in which the pelvic fins played a relatively minor anchoring part, before the emergence of hindlimb-powered propulsion in the interval between Panderichthys and Acanthostega. PMID- 16372008 TI - Dance reveals symmetry especially in young men. AB - Dance is believed to be important in the courtship of a variety of species, including humans, but nothing is known about what dance reveals about the underlying phenotypic--or genotypic--quality of the dancer. One measure of quality in evolutionary studies is the degree of bodily symmetry (fluctuating asymmetry, FA), because it measures developmental stability. Does dance quality reveal FA to the observer and is the effect stronger for male dancers than female? To answer these questions, we chose a population that has been measured twice for FA since 1996 (ref. 9) in a society (Jamaican) in which dancing is important in the lives of both sexes. Motion-capture cameras created controlled stimuli (in the form of videos) that isolated dance movements from all other aspects of visual appearance (including FA), and the same population evaluated these videos for dancing ability. Here we report that there are strong positive associations between symmetry and dancing ability, and these associations were stronger in men than in women. In addition, women rate dances by symmetrical men relatively more positively than do men, and more-symmetrical men value symmetry in women dancers more than do less-symmetrical men. In summary, dance in Jamaica seems to show evidence of sexual selection and to reveal important information about the dancer. PMID- 16372009 TI - Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus. AB - Aspergillus fumigatus is exceptional among microorganisms in being both a primary and opportunistic pathogen as well as a major allergen. Its conidia production is prolific, and so human respiratory tract exposure is almost constant. A. fumigatus is isolated from human habitats and vegetable compost heaps. In immunocompromised individuals, the incidence of invasive infection can be as high as 50% and the mortality rate is often about 50% (ref. 2). The interaction of A. fumigatus and other airborne fungi with the immune system is increasingly linked to severe asthma and sinusitis. Although the burden of invasive disease caused by A. fumigatus is substantial, the basic biology of the organism is mostly obscure. Here we show the complete 29.4-megabase genome sequence of the clinical isolate Af293, which consists of eight chromosomes containing 9,926 predicted genes. Microarray analysis revealed temperature-dependent expression of distinct sets of genes, as well as 700 A. fumigatus genes not present or significantly diverged in the closely related sexual species Neosartorya fischeri, many of which may have roles in the pathogenicity phenotype. The Af293 genome sequence provides an unparalleled resource for the future understanding of this remarkable fungus. PMID- 16372010 TI - Genome sequencing and analysis of Aspergillus oryzae. AB - The genome of Aspergillus oryzae, a fungus important for the production of traditional fermented foods and beverages in Japan, has been sequenced. The ability to secrete large amounts of proteins and the development of a transformation system have facilitated the use of A. oryzae in modern biotechnology. Although both A. oryzae and Aspergillus flavus belong to the section Flavi of the subgenus Circumdati of Aspergillus, A. oryzae, unlike A. flavus, does not produce aflatoxin, and its long history of use in the food industry has proved its safety. Here we show that the 37-megabase (Mb) genome of A. oryzae contains 12,074 genes and is expanded by 7-9 Mb in comparison with the genomes of Aspergillus nidulans and Aspergillus fumigatus. Comparison of the three aspergilli species revealed the presence of syntenic blocks and A. oryzae specific blocks (lacking synteny with A. nidulans and A. fumigatus) in a mosaic manner throughout the genome of A. oryzae. The blocks of A. oryzae-specific sequence are enriched for genes involved in metabolism, particularly those for the synthesis of secondary metabolites. Specific expansion of genes for secretory hydrolytic enzymes, amino acid metabolism and amino acid/sugar uptake transporters supports the idea that A. oryzae is an ideal microorganism for fermentation. PMID- 16372011 TI - NMDA receptors are expressed in oligodendrocytes and activated in ischaemia. AB - Glutamate-mediated damage to oligodendrocytes contributes to mental or physical impairment in periventricular leukomalacia (pre- or perinatal white matter injury leading to cerebral palsy), spinal cord injury, multiple sclerosis and stroke. Unlike neurons, white matter oligodendrocytes reportedly lack NMDA (N-methyl-d aspartate) receptors. It is believed that glutamate damages oligodendrocytes, especially their precursor cells, by acting on calcium-permeable AMPA (alpha amino-3-hydroxy-5-methyl-4-isoxazole propionic acid)/kainate receptors alone or by reversing cystine-glutamate exchange and depriving cells of antioxidant protection. Here we show that precursor, immature and mature oligodendrocytes in the white matter of the cerebellum and corpus callosum exhibit NMDA-evoked currents, mediated by receptors that are blocked only weakly by Mg2+ and that may contain NR1, NR2C and NR3 NMDA receptor subunits. NMDA receptors are present in the myelinating processes of oligodendrocytes, where the small intracellular space could lead to a large rise in intracellular ion concentration in response to NMDA receptor activation. Simulating ischaemia led to development of an inward current in oligodendrocytes, which was partly mediated by NMDA receptors. These results point to NMDA receptors of unusual subunit composition as a potential therapeutic target for preventing white matter damage in a variety of diseases. PMID- 16372012 TI - NMDA receptors are expressed in developing oligodendrocyte processes and mediate injury. AB - Injury to oligodendrocyte processes, the structures responsible for myelination, is implicated in many forms of brain disorder. Here we show NMDA (N-methyl-D aspartate) receptor subunit expression on oligodendrocyte processes, and the presence of NMDA receptor subunit messenger RNA in isolated white matter. NR1, NR2A, NR2B, NR2C, NR2D and NR3A subunits showed clustered expression in cell processes, but NR3B was absent. During modelled ischaemia, NMDA receptor activation resulted in rapid Ca2+-dependent detachment and disintegration of oligodendroglial processes in the white matter of mice expressing green fluorescent protein (GFP) specifically in oligodendrocytes (CNP-GFP mice). This effect occurred at mouse ages corresponding to both the initiation and the conclusion of myelination. NR1 subunits were found mainly in oligodendrocyte processes, whereas AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid)/kainate receptor subunits were mainly found in the somata. Consistent with this observation, injury to the somata was prevented by blocking AMPA/kainate receptors, and preventing injury to oligodendroglial processes required the blocking of NMDA receptors. The presence of NMDA receptors in oligodendrocyte processes explains why previous studies that have focused on the somata have not detected a role for NMDA receptors in oligodendrocyte injury. These NMDA receptors bestow a high sensitivity to acute injury and represent an important new target for drug development in a variety of brain disorders. PMID- 16372013 TI - WUSCHEL controls meristem function by direct regulation of cytokinin-inducible response regulators. AB - Plants continuously maintain pools of totipotent stem cells in their apical meristems from which elaborate root and shoot systems are produced. In Arabidopsis thaliana, stem cell fate in the shoot apical meristem is controlled by a regulatory network that includes the CLAVATA (CLV) ligand-receptor system and the homeodomain protein WUSCHEL (WUS). Phytohormones such as auxin and cytokinin are also important for meristem regulation. Here we show a mechanistic link between the CLV/WUS network and hormonal control. WUS, a positive regulator of stem cells, directly represses the transcription of several two-component ARABIDOPSIS RESPONSE REGULATOR genes (ARR5, ARR6, ARR7 and ARR15), which act in the negative-feedback loop of cytokinin signalling. These data indicate that ARR genes might negatively influence meristem size and that their repression by WUS might be necessary for proper meristem function. Consistent with this hypothesis is our observation that a mutant ARR7 allele, which mimics the active, phosphorylated form, causes the formation of aberrant shoot apical meristems. Conversely, a loss-of-function mutation in a maize ARR homologue was recently shown to cause enlarged meristems. PMID- 16372014 TI - Double chromodomains cooperate to recognize the methylated histone H3 tail. AB - Chromodomains are modules implicated in the recognition of lysine-methylated histone tails and nucleic acids. CHD (for chromo-ATPase/helicase-DNA-binding) proteins regulate ATP-dependent nucleosome assembly and mobilization through their conserved double chromodomains and SWI2/SNF2 helicase/ATPase domain. The Drosophila CHD1 localizes to the interbands and puffs of the polytene chromosomes, which are classic sites of transcriptional activity. Other CHD isoforms (CHD3/4 or Mi-2) are important for nucleosome remodelling in histone deacetylase complexes. Deletion of chromodomains impairs nucleosome binding and remodelling by CHD proteins. Here we describe the structure of the tandem arrangement of the human CHD1 chromodomains, and its interactions with histone tails. Unlike HP1 and Polycomb proteins that use single chromodomains to bind to their respective methylated histone H3 tails, the two chromodomains of CHD1 cooperate to interact with one methylated H3 tail. We show that the human CHD1 double chromodomains target the lysine 4-methylated histone H3 tail (H3K4me), a hallmark of active chromatin. Methylammonium recognition involves two aromatic residues, not the three-residue aromatic cage used by chromodomains of HP1 and Polycomb proteins. Furthermore, unique inserts within chromodomain 1 of CHD1 block the expected site of H3 tail binding seen in HP1 and Polycomb, instead directing H3 binding to a groove at the inter-chromodomain junction. PMID- 16372016 TI - Epidemiology - identifying the causes and preventability of cancer? AB - It has been almost 25 years since Doll and Peto performed their landmark analysis of epidemiological data to identify the causes of cancers and possible modes of cancer prevention. Since then, there have been many additional studies of cancer incidence using various epidemiological techniques. These studies revealed expanded opportunities for cancer prevention through approaches that include vaccination, increased physical activity, weight control and avoidance of post menopausal hormone therapy. PMID- 16372017 TI - Predicting the future burden of cancer. AB - As observations in the past do not necessarily hold into the future, predicting future cancer occurrence is fraught with uncertainty. Nevertheless, predictions can aid health planners in allocating resources and allow scientists to explore the consequence of interventions aimed at reducing the impact of cancer. Simple statistical models have been refined over the past few decades and often provide reasonable predictions when applied to recent trends. Intrinsic to their interpretation, however, is an understanding of the forces that drive time trends. We explain how and why cancer predictions are made, with examples to illustrate the concepts in practice. PMID- 16372018 TI - KLF4, p21 and context-dependent opposing forces in cancer. AB - Kruppel-like factors are transcriptional regulators that influence several cellular functions, including proliferation. Recent studies have shown that one family member, KLF4, can function both as a tumour suppressor and an oncogene. The ability of KLF4 to affect the levels of expression of the cell-cycle regulator p21 seems to be involved, in that this protein might function as a switch that determines the outcome of KLF4 signalling. Is this role of p21 restricted to KLF4, or does p21 represent a nodal point for signals from multiple other factors with opposing functions in cancer? PMID- 16372019 TI - NMDA receptors mediate calcium accumulation in myelin during chemical ischaemia. AB - Central nervous system myelin is a specialized structure produced by oligodendrocytes that ensheaths axons, allowing rapid and efficient saltatory conduction of action potentials. Many disorders promote damage to and eventual loss of the myelin sheath, which often results in significant neurological morbidity. However, little is known about the fundamental mechanisms that initiate myelin damage, with the assumption being that its fate follows that of the parent oligodendrocyte. Here we show that NMDA (N-methyl-d-aspartate) glutamate receptors mediate Ca2+ accumulation in central myelin in response to chemical ischaemia in vitro. Using two-photon microscopy, we imaged fluorescence of the Ca2+ indicator X-rhod-1 loaded into oligodendrocytes and the cytoplasmic compartment of the myelin sheath in adult rat optic nerves. The AMPA (alpha-amino 3-hydroxy-5-methyl-4-isoxazole propionic acid)/kainate receptor antagonist NBQX completely blocked the ischaemic Ca2+ increase in oligodendroglial cell bodies, but only modestly reduced the Ca2+ increase in myelin. In contrast, the Ca2+ increase in myelin was abolished by broad-spectrum NMDA receptor antagonists (MK 801, 7-chlorokynurenic acid, d-AP5), but not by more selective blockers of NR2A and NR2B subunit-containing receptors (NVP-AAM077 and ifenprodil). In vitro ischaemia causes ultrastructural damage to both axon cylinders and myelin. NMDA receptor antagonism greatly reduced the damage to myelin. NR1, NR2 and NR3 subunits were detected in myelin by immunohistochemistry and immunoprecipitation, indicating that all necessary subunits are present for the formation of functional NMDA receptors. Our data show that the mature myelin sheath can respond independently to injurious stimuli. Given that axons are known to release glutamate, our finding that the Ca2+ increase was mediated in large part by activation of myelinic NMDA receptors suggests a new mechanism of axo-myelinic signalling. Such a mechanism may represent a potentially important therapeutic target in disorders in which demyelination is a prominent feature, such as multiple sclerosis, neurotrauma, infections (for example, HIV encephalomyelopathy) and aspects of ischaemic brain injury. PMID- 16372020 TI - Structural basis for Duffy recognition by the malaria parasite Duffy-binding-like domain. AB - Molecular processes that govern pathogenic features of erythrocyte invasion and cytoadherence in malaria are reliant on Plasmodium-specific Duffy-binding-like domains (DBLs). These cysteine-rich modules recognize diverse host cell-surface receptors during pathogenesis. DBLs of parasite erythrocyte-binding proteins mediate invasion, and those from the antigenically variant P. falciparum erythrocyte membrane protein 1 (PfEMP1) have been implicated in cytoadherence. The simian and human malarial parasites, P. knowlesi and P. vivax, invade human erythrocytes exclusively through the host DARC receptor (Duffy antigen receptor for chemokines). Here we present the crystal structure of the P. knowlesi DBL domain (Pkalpha-DBL), which binds to DARC during invasion of human erythrocytes. Pkalpha-DBL retains the overall fold observed in DBLs from P. falciparum erythrocyte-binding antigen (EBA)-175 (ref. 4). Mapping the residues that have previously been implicated in binding highlights a fairly flat but exposed site for DARC recognition in subdomain 2 of Pkalpha-DBL; this is in sharp contrast to receptor recognition by EBA-175 (ref. 4). In Pkalpha-DBL, the residues that contact DARC and the clusters of residues under immune pressure map to opposite surfaces of the DBL, and suggest a possible mechanism for immune evasion by P. vivax. Our comparative structural analysis of Pkalpha-DBL and P. falciparum EBA 175 provides a framework for the understanding of malaria parasite DBLs, and may affect the development of new prophylactic and therapeutic strategies. PMID- 16372021 TI - Origins of extrinsic variability in eukaryotic gene expression. AB - Variable gene expression within a clonal population of cells has been implicated in a number of important processes including mutation and evolution, determination of cell fates and the development of genetic disease. Recent studies have demonstrated that a significant component of expression variability arises from extrinsic factors thought to influence multiple genes simultaneously, yet the biological origins of this extrinsic variability have received little attention. Here we combine computational modelling with fluorescence data generated from multiple promoter-gene inserts in Saccharomyces cerevisiae to identify two major sources of extrinsic variability. One unavoidable source arising from the coupling of gene expression with population dynamics leads to a ubiquitous lower limit for expression variability. A second source, which is modelled as originating from a common upstream transcription factor, exemplifies how regulatory networks can convert noise in upstream regulator expression into extrinsic noise at the output of a target gene. Our results highlight the importance of the interplay of gene regulatory networks with population heterogeneity for understanding the origins of cellular diversity. PMID- 16372022 TI - Gamma-band synchronization in visual cortex predicts speed of change detection. AB - Our capacity to process and respond behaviourally to multiple incoming stimuli is very limited. To optimize the use of this limited capacity, attentional mechanisms give priority to behaviourally relevant stimuli at the expense of irrelevant distractors. In visual areas, attended stimuli induce enhanced responses and an improved synchronization of rhythmic neuronal activity in the gamma frequency band (40-70 Hz). Both effects probably improve the neuronal signalling of attended stimuli within and among brain areas. Attention also results in improved behavioural performance and shortened reaction times. However, it is not known how reaction times are related to either response strength or gamma-band synchronization in visual areas. Here we show that behavioural response times to a stimulus change can be predicted specifically by the degree of gamma-band synchronization among those neurons in monkey visual area V4 that are activated by the behaviourally relevant stimulus. When there are two visual stimuli and monkeys have to detect a change in one stimulus while ignoring the other, their reactions are fastest when the relevant stimulus induces strong gamma-band synchronization before and after the change in stimulus. This enhanced gamma-band synchronization is also followed by shorter neuronal response latencies on the fast trials. Conversely, the monkeys' reactions are slowest when gamma-band synchronization is high in response to the irrelevant distractor. Thus, enhanced neuronal gamma-band synchronization and shortened neuronal response latencies to an attended stimulus seem to have direct effects on visually triggered behaviour, reflecting an early neuronal correlate of efficient visuo-motor integration. PMID- 16372023 TI - Ethnic differences in nicotine metabolic rate among New Zealanders. AB - AIMS: To estimate (a) the prevalence of gene variants associated with slow nicotine metabolism in the general Maori population and (b) nicotine intake and metabolic rate in Maori and European smokers. METHODS: The procedure involved (a) genotyping 85 Maori participants for cytochrome P-450 2A6 (CYP2A6) gene variants, which are associated with reduced nicotine metabolic rate (ie CYP2A6*9 and *4); and (b) measuring salivary cotinine (COT) and trans-3'-hydroxycotinine (3-HC) as biomarkers of nicotine intake and metabolic rate in 12 female smokers from the Hawke's Bay Region (6 Maori and 6 European). RESULTS: (a) The frequencies of the slow nicotine metabolising variants, CYP2A6*9 and *4, were significantly higher in Maori compared to European (p<0.01). Indeed, the prevalence of the CYP2A6*9 variant in these Maori was among the highest in the world (approximately 20%). (b) In smokers, the Maori group had approximately 35% lower 3-HC:COT ratios indicating a reduced metabolic rate, as well as 2-fold lower cotinine levels per cigarette smoked, indicating reduced nicotine intake (p<0.05). The CYP2A6*9 allele was significantly more frequent in Maori smokers (70%) compared to Europeans (30%), p=0.03. CONCLUSIONS: The findings of this study provide evidence that Maori are genetically slower nicotine metabolisers compared to Europeans. Although more research is required, this study may help explain ethnic differences in smoking initiation and may also have important implications for smoking cessation programs - since metabolic differences between groups with varying ancestry implies that different optimal dosages of nicotine replacement therapy may be required for successful quitting. PMID- 16372024 TI - Evidence, economics, and emotions: the case for temozolomide. AB - Temozolomide, given as part of first line therapy in the treatment of grade IV astrocytoma, has been shown to improve survival in the short term. The financial cost of the treatment is considerable in New Zealand. This drug provides a good example in the field of oncology of a modern expensive pharmaceutical being a clear improvement over its cheaper predecessors, but it raises the question of what price should be paid to prolong survival in an incurable illness? PMID- 16372025 TI - Massive haemorrhage caused by a bone marrow aspirate and trephine (BMAT) procedure in a uraemic patient. PMID- 16372026 TI - Post-traumatic haematoma: a red herring to something more sinister. PMID- 16372027 TI - Medical image. A rare complication of port-a-cath use. PMID- 16372028 TI - The contribution of causes of death to socioeconomic inequalities in child mortality: New Zealand 1981-1999. AB - BACKGROUND: Socioeconomic inequalities in all-cause child mortality exist in New Zealand; however the inequalities in cause-specific mortality have not been examined. This study examines child mortality inequality by household income between 1981 and 1999, by cause of death. METHODS: Data was used from a record linkage study of census and mortality records of all New Zealand children aged 0 14 years on census night 1981, 1986, 1991, 1996 followed up for 3 years for specific causes of mortality between ages 1-14 years. All cohorts were combined to calculate mortality rates, rate ratios, and rates differences for each cause of death. RESULTS: Socioeconomic differences in child mortality (low income compared to high income) were observed for injury (non road traffic) (RR 1.87, 1.35 to 2.58), road traffic injury (RR 1.36, 1.01 to 1.82), and 'other' causes of death (RR 1.81, 1.32 to 2.47). 'Other' and non-road traffic injury deaths together contributed 70% of the total gap in child mortality between the rich and the poor. CONCLUSIONS: Socioeconomic differences existed across most broad causes of child death. The major contributors to mortality inequality are diverse, suggesting that the similar distal causes of inequality (e.g. poverty) play out through a myriad of proximal causes. Fortunately there appears to be some scope for policymakers to modify some of the proximal and distal causes of these inequalities. PMID- 16372029 TI - Aetiology and pathogenesis of chronic fatigue syndrome: a review. AB - Chronic fatigue syndrome (CFS) is a debilitating disease of uncertain aetiology that is characterised by unexplained, severe fatigue associated with a number of typical symptoms. This paper reviews the scientific literature related to current theories about the aetiology and pathogenesis of CFS by focussing on what appear to be the four most significant aspects in the development and perpetuation of this disease: the role of infectious agents as well as immunological, neuroendocrine, and psychiatric factors. A multifactorial model for the aetiology of CFS, which includes and draws together these four aspects, is proposed; and suggestions are offered regarding approaches to the diagnosis and treatment of this disease. PMID- 16372030 TI - Attitudes of hospital medical practitioners to the mandatory reporting of professional misconduct. AB - BACKGROUND: New legislation now requires doctors to report unfit colleagues to the Medical Council of New Zealand. However, little research has examined the attitudes and willingness of doctors to report errant colleagues. AIM: To examine the attitudes of a range of hospital-based medical practitioners towards the mandatory reporting of colleagues. METHODS: House staff, registrars, and consultants at two major tertiary teaching hospitals in New Zealand were surveyed using a written questionnaire. Doctors were asked to state their level of agreement with a series of statements and to make responses to three hypothetical scenarios: an alcohol impaired practitioner; a senior colleague with recent behavioural change; and a surgeon expressing racist views. RESULTS: Responses were received from 52% of medical staff at the two hospitals. Respondents were consultants (52%), registrars (39%), and house staff (9%). Most (98%) respondents agreed that all doctors make clinical errors and there was a need for open discussion about error. The majority (80%) also accepted that doctors were responsible for the actions of colleagues and agreed that they would act if a colleague was failing to achieve professional standards. Only 45% agreed with the mandatory reporting of error. The responses to three scenarios illustrate some variability in how different groups of doctors would address the behaviour of an errant colleague. CONCLUSIONS: Although most hospital doctors accept they should act if a colleague is falling below professional standards, there is only limited support for mandatory reporting; instead, doctors may prefer to consult senior colleagues about an errant colleague or sometimes counsel the practitioner themselves. PMID- 16372031 TI - Secondhand smoke in New Zealand homes and cars: exposure, attitudes, and behaviours in 2004. AB - AIMS: To assess exposure to secondhand smoke (SHS) in New Zealand homes and cars and to describe attitudes and behaviours that relate to establishing smokefree settings. METHODS: In 2004, a nationwide telephone survey randomly selected and interviewed 2731 respondents. This sample was weighted to represent the national population and was comprised of a general adult population sample (n=1507), a sample selected from the Maori electoral roll (n=924), and a sample of current smokers and people who had recently quit smoking (n=300). RESULTS: 19.6% of the general population and 42.7% of the Maori sample reported current smoking. Of all current smokers, 47.2% smoked indoors at home and 70.8% smoked in their cars. Maori participants were significantly more likely to be exposed to SHS in their homes than non-Maori participants. There was also some evidence for lower socioeconomic status being related to higher SHS exposure. Extrapolating from the adult exposure data in households with children, it was estimated that 18.9% of children were potentially exposed to SHS indoors at home. However, most respondents (73.6%) lived in homes with total smoking bans. CONCLUSIONS: SHS exposure remains a significant problem especially for Maori and low-income New Zealanders. There is a need for further public health campaigns to increase the prevalence of smokefree homes and cars. PMID- 16372032 TI - Work-related fatal traffic crashes in New Zealand: 1985-1998. AB - AIM: To identify and describe all work-related traffic fatalities in New Zealand between 1985 and 1998 inclusive. METHODS: Potential cases were identified from databases held by three national agencies. The circumstances of the deaths in each fatal incident were reviewed directly from coronial files to determine work relatedness. RESULTS: The rate of work-related fatal injury involving vehicles on a public road was 2.01 per 100,000 workers per year. The rate for worker deaths was 1.11 and that for commuting deaths was 0.89 per 100,000 workers per year. CONCLUSIONS: There is a substantial number of work-related crash fatalities each year and these represent a sizeable portion of the total burden of work-related fatalities. PMID- 16372033 TI - Toxic effects of BZP-based herbal party pills in humans: a prospective study in Christchurch, New Zealand. AB - AIM: This study describes patterns of human toxicity related to the use of 1 benzylpiperazine (BZP)-based 'herbal party pills'. METHODS: From 1 April 2005 to 1 September 2005 all presentations associated with party pill use were captured on a prospective data collection form. RESULTS: There were 61 patients who presented on 80 occasions to the Emergency Department of Christchurch Hospital, New Zealand. Patients with adverse effects took an average of 4.5 tablets/capsules. Patients with mild to moderate toxicity experienced symptoms such as insomnia, anxiety, nausea, vomiting, palpitations, dystonia, and urinary retention. Some adverse reactions persisted up to 24 hours after ingestion. Fifteen toxic seizures were recorded. Two patients suffered life-threatening toxicity with status epilepticus and severe respiratory and metabolic acidosis. CONCLUSIONS: Herbal party pills have been sold without regulation since 2000, and are now widely used by young New Zealanders. The principal ingredient of these pills is 1-benzylpiperazine (BZP). They appear to have a narrow safety margin when used recreationally by some humans, possibly because of intrinsic pharmacodynamic properties, self-dosing variability, or genetic polymorphism. Those with seizure disorders or coronary disease should avoid BZP as should those taking prescription sympathomimetics or anticholinergics. Coingestion with MDMA or amphetamine should also be cautioned against. The results of this study indicate that BZP can cause unpredictable and serious toxicity in some individuals. Furthermore, the results of this study should be carefully considered in any discussion on the legal status of piperazine-based party pills. PMID- 16372034 TI - Widening the lens on child health. PMID- 16372035 TI - The hazards of driving to work. PMID- 16372036 TI - Perceptions of New Zealand adults about complementary and alternative therapies for cancer treatment. AB - AIM: To study perceptions regarding complementary and alternative medicine (CAM) treatments for cancer among adult New Zealanders. METHODS: An anonymous telephone questionnaire that included questions to explore perceptions about CAM and cancer treatment was administered to a randomly selected sample of New Zealanders, 20 years and older. RESULTS: A total of 438 New Zealand adults participated in the survey, out of 689 eligible contacts (68% participation). Less than one-third (28%) agreed with the statement alternative therapy for cancer has an equal or better chance of curing cancer as medical treatment, 34% disagreed, and 38% said they did not know. Most (63%) felt that complementary therapies could be beneficial to people who were also receiving conventional cancer treatment, although only 36% could name one or more such therapies. One-third (32%) said that alternative therapies could be used instead of conventional cancer treatments, but only 16% of the sample could name any alternative therapies. The CAM therapies named most often were nutrition (vitamin and mineral supplements, herbs, and diets) and psychosocial therapies (including positive thinking, spiritual therapies, and relaxation). CONCLUSIONS: There seems to be little consensus about the efficacy of CAM therapies for cancer. New Zealanders may lack information about CAM, or may be withholding judgment because of contradictory messages. PMID- 16372037 TI - Hong Kong case-control study of sudden unexpected infant death. AB - AIM: To document causes of all unexpected child deaths under 2 years of age during a 4-year period (1999-2003), and to identify factors associated with sudden infant death syndrome (SIDS) in Hong Kong. METHODS: The case-control component of the study compared information from SIDS deaths (n=16) with healthy controls (n=223) identified randomly from all births in Hong Kong. Coroner records of all deaths under 2 years of age were later reviewed. RESULTS: SIDS risk factors included prone sleep position, smoking by mother, bedsharing with someone other than the parents, and baby found with head covered. Eighteen deaths were officially classified as SIDS but, on review of the coroner records, there were 33 potential SIDS deaths (many labelled as unascertained/unknown). CONCLUSION: Hong Kong SIDS incidence has fallen from 0.3/1000 (95% CI: 0.18-0.46 in 1987) to 0.16/1000 (95% CI: 0.11-0.22 in 1999-2002). Despite the small number of cases, key SIDS risk factors are shown to be important in this population. Hong Kong needs to take steps to standardise the investigation and management of these deaths and to establish a child mortality review mechanism to provide feedback to the public, to the health authorities, and to health professionals. PMID- 16372038 TI - Statins and myopathy. PMID- 16372039 TI - PHARMAC responds on TNF inhibitors for inflammatory arthritis. PMID- 16372040 TI - Library outreach for health professionals. PMID- 16372041 TI - PHARMAC responds on treatments for pulmonary arterial hypertension. PMID- 16372042 TI - PHARMAC's response on temozolomide and funding costly medicines that prolong life shortly. PMID- 16372043 TI - Manipulation of ovarian follicle development by injecting vascular endothelial growth factor (VEGF) gene. AB - The genetic and molecular mechanisms that control the development of capillary blood vessels during follicular development are beginning to be elucidated. Ovarian follicles contain and produce angiogenic factors that may act alone or in concert to regulate thecal angiogenesis. These factors are ultimately controlled by endocrine, paracrine and autocrine regulation in the ovary. Our recent study indicated that vascular endothelial growth factor (VEGF) plays an important role in the thecal angiogenesis during follicular development. In this review, we focus on the vasculature and the expression of angiogenic factors during follicular development in a mammalian ovary. PMID- 16372044 TI - Follicular dysfunction induced by autoimmunity to zona pellucida. AB - The mammalian zona pellucida is an extracellular matrix that occurs in growing oocytes, ovulated eggs and pre-implantation embryos, and is known to be involved in several important events during ovarian folliculogenesis and fertilization. Since the zona pellucida is formed at an early stage of oocyte growth, circulating antibodies against zona pellucida may impair ovarian function. In this article we discuss whether anti-zona antibodies cause ovarian dysfunction and infertility. The discussion is based on clinical examination and animal experiments including the following approaches: 1/ immunological method using solubilized human zona pellucida detected anti-zona antibody with a high frequency in infertile patients, especially premature ovarian failure syndrome; 2/ in vivo experiment using hamsters showed that some, but not all, animals experienced ovarian failure after immunization with hamster recombinant zona proteins; 3/ in vitro experiment using mouse isolated ovarian follicles showed significant inhibitory effects on follicular growth and oocyte development. We concluded that anti-zona antibody may be involved in causing ovarian failure. PMID- 16372045 TI - Proteomics of boar seminal plasma - current studies and possibility of their application in biotechnology of animal reproduction. AB - Proteomics is critical to identify the properties and functions of proteins involved in the mechanism regulating the male reproductive tract function. This approach is important in male fertility assessment and clinical diagnosis of the physiological state of individual reproductive organs. Proteomics also provides a tool to understand the interactions of seminal plasma proteins with spermatozoa, which could provide a useful model for studying ligand-cell interaction occurring at the sperm cell surface. This review covers a selection of advances in the realm of functional proteomics of boar seminal plasma proteins and is focused on some fundamental proteomic technologies. Also, this review explores key themes in proteomics and their application in animal reproductive techniques. PMID- 16372046 TI - Immunolocalization of androgen receptor in the epididymis of rats with dihydrotestosterone deficiency. AB - Dihydrotestosterone (DHT), 5alpha-reduced metabolite of testosterone, is the most potent androgen in the epididymis. The conversion of T into DHT is carried out by 5alpha-reductase. The activity of 5alpha-reductase type 2, preferentially expressed in the epididymis can be inhibited by a finasteride (a steroid-based specific inhibitor of 5alpha-reductase type 2) which results in DHT deficiency. The aim of the study was to examine the morphology of epididymis and the immunolocalization of an androgen receptor (AR) in the initial segment, caput and cauda epididymis of rats treated with finasteride for 56 days. There were no morphological changes in the morphology of epididymal epithelium in the experimental rats. Immunostainable AR was localized in nuclei of epithelial cells, smooth muscle cells and mainly in the cytoplasm of interstitial cells in the epididymis of control rats. In the epididymis of experimental rats, AR immunostaining was noticed mainly in the cytoplasm of epithelial cells and interstitial cells. The single cells of the initial segment epithelium, basal cells and smooth muscle cells of cauda epididymis showed nuclear AR staining. In conclusion, finasteride affected the expression of the AR in the rat epididymis without changing the morphology of epididymal epithelium. Altered AR expression reflected the hormonal status within the epididymis. PMID- 16372047 TI - In vitro effects of polychlorinated biphenyls (PCBs) on the contractility of bovine myometrium from the periovulatory stage of the estrous cycle. AB - Strips of longitudinal myometrium from cows were obtained on days 19-21 and 1-5 of the estrous cycle and incubated (aerated atmosphere; 4 degrees C; 24, 48 or 72 h) with a mixture of PCBs Aroclor (Ar) 1248 or with one of three PCBs (77, 126 or 153), all at doses of 10 or 100 ng/ml. The force and frequency of spontaneous and oxytocin (OT; 10(-7)M)-stimulated contractions of each strip was registered by means of HSE Schuler Organbath. Contractions of myometrial strips in the presence and absence of PCBs were observed after 24, 48 and 72 h of incubation. All PCBs significantly affected myometrial contractions. A mixture of PCBs increased the spontaneous force of contractions after 24 h but decreased after 48 h. Individual congeners of PCB also amplified the force of contractions and in most cases this effect was dose-dependent. Response of myometrium to PCB-126 and PCB-153 or PCB 77 appeared after 24 h or 48 h of incubation. Incubation of myometrial strips with PCB congeners markedly amplified OT-stimulated contractions. This effect was less evident when tissue was pre-treated with a higher dose of PCBs. Pre treatment with estrogen-like PCB-153 increased the spontaneous and OT-evoked frequency of myometrial contractions from days 19-21. The spontaneous force of myometrial strips' contractions as well as the effects evoked by PCBs and OT was higher before than after ovulation. In summary, PCBs affected both the force and frequency of uterine contractions. Thus, it can be concluded that PCBs may impair both ovum fertilization and blastocyst implantation in cows. PMID- 16372048 TI - Effect of polychlorinated biphenyls (PCBs) on basal and OT-stimulated calcium concentrations in myometrial cells in cows. AB - We investigated the effect of PCB-77, -126 or -153 (10 or 100 ng/ml) on free intracellular calcium concentrations([Ca2+]i) in bovine myometrial cells from days 1-5 of the estrous cycle. Cells were incubated with or without PCBs for 48 h (38 degrees C, aerated atmosphere) and thereafter [Ca2+]i was measured by means of fluorescent calcium indicator Fura-2. PCBs increased basal concentrations of [Ca2+]i measured before oxytocin (OT) challenge. The increase in [Ca2+]i in cells incubated with PCBs and challenged with OT was inhibited or delayed when compared to control OT-stimulated cells (p<0.05). The applied doses of PCBs did not affect viability of myometrial cells. In conclusion, the influence of PCBs upon intracellular calcium mobilization in myometrial cells impaired the bovine uterus contractility. PMID- 16372049 TI - Study on the role of a sperm membrane-associated antigen in canine fertilization. AB - The Mab 4B12 produced by us against capacitated boar spermatozoa was found to recognize a protein located in the acrosome portion of capacitated boar spermatozoa which is shared by different animal species, dogs included. It was shown that Mab 4B12 might affect fertilizing ability in vitro of boar spermatozoa. Using indirect immunofluorescence (IIF) test, we provide evidence here that Mab 4B12 stained the acrosome of the capacitated but not of the ejaculated and acrosome-reacted canine spermatozoa. The biological experiments using hemizona assay functional test in this study provide evidence supporting the involvement of Mab 4B12 corresponding antigen in the functional steps required for canine sperm-zona pellucida binding. These results together with the data on cell and tissue specificity of the 4B12 antigen suggest its contraceptive potential for canine fertilization. PMID- 16372050 TI - Imaging of serotonin mechanisms in epilepsy. AB - Advances in positron emission tomography (PET) techniques have allowed the measurement and imaging of neurotransmitter synthesis, transport, and receptor binding to be performed in vivo. With regard to epileptic disorders, imaging of neurotransmitter systems not only assists in the identification of epileptic foci for surgical treatment, but also provides insights into the basic mechanisms of human epilepsy. Recent investigative interest in epilepsy has focused on PET imaging of tryptophan metabolism, via the serotonin and kynurenine pathways, as well as on imaging of serotonin receptors. This review summarizes advances in PET imaging and how these techniques can be applied clinically for epilepsy treatment. PMID- 16372051 TI - The terminal man--from science fiction to therapy. PMID- 16372052 TI - Periictal dynamic changes in benzodiazepine receptors. PMID- 16372053 TI - Anatomical and behavioral effects of in utero exposure to antiepileptic drugs. PMID- 16372054 TI - Pregabalin: the next new thing. PMID- 16372055 TI - Angelman syndrome: need for further illumination in the theater of the happy puppet. PMID- 16372056 TI - Sudden death and epilepsy. PMID- 16372058 TI - When newborn neurons stray. PMID- 16372057 TI - Update on the neurobiology of alcohol withdrawal seizures. AB - Abrupt cessation of alcohol intake after prolonged heavy drinking may trigger alcohol withdrawal seizures. Generalized tonic-clonic seizures are the most characteristic and severe type of seizure that occur in this setting. Generalized seizures also occur in rodent models of alcohol withdrawal. In these models, the withdrawal seizures are triggered by neuronal networks in the brainstem, including the inferior colliculus; similar brainstem mechanisms may contribute to alcohol withdrawal seizures in humans. Alcohol causes intoxication through effects on diverse ion channels and neurotransmitter receptors, including GABA(A) receptors--particularly those containing delta subunits that are localized extrasynaptically and mediate tonic inhibition--and N-methyl-D-aspartate (NMDA) receptors. Alcohol dependence results from compensatory changes during prolonged alcohol exposure, including internalization of GABA(A) receptors, which allows adaptation to these effects. Withdrawal seizures are believed to reflect unmasking of these changes and may also involve specific withdrawal-induced cellular events, such as rapid increases in alpha4 subunit-containing GABA(A) receptors that confer reduced inhibitory function. Optimizing approaches to the prevention of alcohol withdrawal seizures requires an understanding of the distinct neurobiologic mechanisms that underlie these seizures. PMID- 16372059 TI - Hypoxia results in GABAergic channelopathy. PMID- 16372060 TI - Pharmacogenetics of epilepsy: one step forward? PMID- 16372061 TI - Chronic cortical injury and epileptogensis: a possible role for intracellular chloride homeostasis. PMID- 16372062 TI - Targeting the brain: focal delivery of natural anticonvulsant molecules. PMID- 16372063 TI - A possible causative role for blood-brain barrier failure and reactive astrocytosis in acquired epilepsy. PMID- 16372065 TI - Nanotechnology for membranes, filters and sieves. A series of mini-reviews covering new trends in fundamental and applied research, and potential applications of miniaturised technologies. PMID- 16372066 TI - Magnetism and microfluidics. AB - Magnetic forces are now being utilised in an amazing variety of microfluidic applications. Magnetohydrodynamic flow has been applied to the pumping of fluids through microchannels. Magnetic materials such as ferrofluids or magnetically doped PDMS have been used as valves. Magnetic microparticles have been employed for mixing of fluid streams. Magnetic particles have also been used as solid supports for bioreactions in microchannels. Trapping and transport of single cells are being investigated and recently, advances have been made towards the detection of magnetic material on-chip. The aim of this review is to introduce and discuss the various developments within the field of magnetism and microfluidics. PMID- 16372067 TI - Implementation of radiotelemetry in a lab-in-a-pill format. AB - A miniaturised lab-in-a-pill device has been produced incorporating a temperature and pH sensor with wireless communication using the 433.92 MHz ISM band. The device has been designed in order to enable real time in situ measurements in the gastrointestinal (GI) tract, and accordingly, issues concerning the resolution and accuracy of the data, and the lifetime of the device have been considered. The sensors, which will measure two key parameters reflecting the physiological environment in the GI (as indicators for disease) were both controlled by an application specific integrated circuit (ASIC). The data were sampled at 10-bit resolution prior to communication off chip as a single interleaved data stream. This incorporated a power saving serial bitstream data compression algorithm that was found to extend the service lifetime of the pill by 70%. An integrated on-off keying (OOK) radio transmitter was used to send the signal to a local receiver (base station), prior to acquisition on a computer. A permanent magnet was also incorporated in the device to enable non-visual tracking of the system. We report on the implementation of this device, together with an initial study sampling from within the porcine GI tract, showing that measurements from the lab-on-a pill, in situ, was within 90% of literature values. PMID- 16372068 TI - A disposable microfluidic cassette for DNA amplification and detection. AB - A pneumatically driven, disposable, microfluidic cassette comprised of a polymerase chain reaction (PCR) thermal cycler, an incubation chamber to label PCR amplicons with up-converting phosphor (UPT) reporter particles, conduits, temperature-activated, normally closed hydrogel valves, and a lateral flow strip, was constructed and tested. The hydrogel valves, which were opened and closed with the aid of electrically controlled thermoelectric units, provided a simple means to seal the PCR reactor and suppress bubble formation. The hydrogel-based flow control was electronically addressable, leakage-free, and biocompatible. To test the device, a solution laden with genomic DNA isolated from B. cereus was introduced into the microfluidic cassette and a specific 305 bp fragment was amplified. The PCR amplicons were labelled with the phosphor (UPT) reporter particles, applied to the lateral flow strip, bound to pre-immobilized ligands, and detected with an IR laser that scanned the lateral flow strip and excited the phosphor (UPT) particles that, in turn, emitted light in the visible spectrum. The UPT particles do not bleach, they provide a permanent record, and they readily facilitate the filtering of background noise. The cassette described herein will be used for rapid testing at the point of care. PMID- 16372069 TI - Soft lithographic patterning of supported lipid bilayers onto a surface and inside microfluidic channels. AB - We present simple soft lithographic methods for patterning supported lipid bilayer (SLB) membranes onto a surface and inside microfluidic channels. Micropatterns of polyethylene glycol (PEG)-based polymers were fabricated on glass substrates by microcontact printing or capillary moulding. The patterned PEG surfaces have shown 97 +/- 0.5% reduction in lipid adsorption onto two dimensional surfaces and 95 +/- 1.2% reduction inside microfluidic channels in comparison to glass control. Atomic force microscopy measurements indicated that the deposition of lipid vesicles led to the formation of SLB membranes by vesicle fusion due to hydrophilic interactions with the exposed substrate. Furthermore, the functionality of the patterned SLBs was tested by measuring the binding interactions between biotin (ligand)-labeled lipid bilayer and streptavidin (receptor). SLB arrays were fabricated with spatial resolution down to approximately 500 nm on flat substrate and approximately 1 microm inside microfluidic channels, respectively. PMID- 16372070 TI - Microfluidic discharge-based optical sources for detection of biochemicals. AB - This paper reports a discharge-based optical source for fluorescence of biochemicals in microfluidic systems. Its efficacy is demonstrated using a stacked microchip that integrates a microfluidic wavelength-tunable optical source, a biochemical sample reservoir and optical filters. It is shown to excite fluorescence in l-tryptophan and DNA samples labeled by SYBR green dye. The discharge is struck in ambient air, between a metal anode and a cathode cavity that is filled with an aqueous solution, which is doped with a metal salt selected for its emission characteristics. The characteristic line spectra, which arise from energetic transitions of the metal ions that are sputtered into the glow region of the discharge, are optically filtered and guided to the biochemical sample that resides in a separate on-chip reservoir. For DNA fluorescence, a barium chloride solution is used to emit light at 454 and 493 nm. For tryptophan fluorescence, the cathode contains lead (ii) nitrate solution to provide a 280 nm emission. The resulting fluorescence from the DNA and tryptophan samples is compared to reference data. This technique can also be used to excite other fluorophores by using appropriately doped liquid cathodes having the desired emission characteristics. PMID- 16372071 TI - Fabrication and characterization of poly(methylmethacrylate) microfluidic devices bonded using surface modifications and solvents. AB - The fabrication of polymer microchips allows inexpensive, durable, high throughput and disposable devices to be made. Poly(methylmethacrylate) (PMMA) microchips have been fabricated by hot embossing microstructures into the substrate followed by bonding a cover plate. Different surface modifications have been examined to enhance substrate and cover plate adhesion, including: air plasma treatment, and both acid catalyzed hydrolysis and aminolysis of the acrylate to yield carboxyl and amine-terminated PMMA surfaces. Unmodified PMMA surfaces were also studied. The substrate and cover plate adhesion strengths were found to increase with the hydrophilicity of the PMMA surface and reached a peak at 600 kN m(-2) for plasma treated PMMA. A solvent assisted system has also been designed to soften less than 50 nm of the surface of PMMA during bonding, while still maintaining microchannel integrity. The extent to which both surface modifications and solvent treatment affected the adhesion of the substrate to the cover plate was examined using nanoindentation methods. The solvent bonding system greatly increased the adhesion strengths for both unmodified and modified PMMA, with a maximum adhesion force of 5500 kN m(-2) achieved for unmodified PMMA substrates. The bond strength decreased with increasing surface hydrophilicity after solvent bonding, a trend that was opposite to what was observed for non solvent thermal bonding. PMID- 16372072 TI - Fluid mixing in planar spiral microchannels. AB - Mixing of fluids at the microscale poses a variety of challenges, many of which arise from the fact that molecular diffusion is the dominant transport mechanism in the laminar flow regime. While considerable progress has been made toward developing strategies to achieve improved mixing in microfluidic systems, many of these techniques introduce additional complexity to device fabrication and/or operation processes. In this work, we explore the use of compact spiral-shaped flow geometries designed to achieve efficient mixing in a format that can be constructed using a single planar soft lithography step without the need for multilayer alignment. A series of 150 microm-wide by 29 microm-tall channels were constructed, each of which incorporated a series of spiral shaped sections arrayed along the flow path. Five spiral designs with varying channel lengths were investigated, and mixing studies were carried out at flow rates corresponding to Reynolds numbers ranging from 0.02 to 18.6. Under appropriate conditions, transverse Dean flows are induced that augment diffusive transport and promote enhanced mixing in considerably shorter downstream distances as compared with conventional planar straight channel designs. Mixing efficiency can be further enhanced by incorporating expansion vortex effects via abrupt changes in cross-sectional area along the flow path. PMID- 16372073 TI - Microfluidic diffusive filter for apheresis (leukapheresis). AB - Apheresis is a procedure used to fractionate whole blood into its individual components. Following fractionation, the desired component is isolated and the remaining blood in many cases is returned to the donor. Leukapheresis is one type of apheresis where leukocytes (white blood cells) are selectively removed. This procedure is commonly used for blood transfusions to remove donor leukocytes from being transferred to the recipient. Apheresis also has several therapeutic applications. In this manuscript we discuss the design, fabrication and testing of a continuous flow diffusive filter, fabricated using simple soft lithographic techniques for depletion of leukocytes. This device employs micro sieves that exploit the size and shape difference between the different cell types to obtain depletion of leukocytes from whole blood. Currently, conventional apheresis methods like centrifugation or fiber mesh filtration are commonly used. A theoretical model was developed to determine the optimal shape of the diffuser to ensure that the volumetric flow through individual sieve elements is equal. This device was designed to serve as a passive device that does not require any external manipulation. Results show that for the given device design, isolation of approximately 50% of the inlet erythrocytes (red blood cells), along with depletion of >97% of the inlet leukocytes is possible at a flow rate of 5 microl min(-1). Simple modifications to the geometry and dimensions of the sieves can be made to obtain isolation of plasma. PMID- 16372074 TI - Monitoring of fluid motion in a micromixer by dynamic NMR microscopy. AB - The velocity distribution of liquid flowing in a commercial micromixer has been determined directly by using pulsed-field gradient NMR. Velocity maps with a spatial resolution of 29 microm x 43 microm were obtained by combining standard imaging gradient units with a homebuilt rectangular surface coil matching the mixer geometry. The technique provides access to mixers and reactors of arbitrary shape regardless of optical transparency. Local heterogeneities in the signal intensity and the velocity pattern were found and serve to investigate the quality and functionality of a micromixer, revealing clogging and inhomogeneous flow distributions. PMID- 16372075 TI - Digital microfluidics using soft lithography. AB - Although microfluidic chips have demonstrated basic functionality for single applications, performing varied and complex experiments on a single device is still technically challenging. While many groups have implemented control software to drive the pumps, valves, and electrodes used to manipulate fluids in microfluidic devices, a new level of programmability is needed for end users to orchestrate their own unique experiments on a given device. This paper presents an approach for programmable and scalable control of discrete fluid samples in a polydimethylsiloxane (PDMS) microfluidic system using multiphase flows. An immiscible fluid phase is utilized to separate aqueous samples from one another, and a novel "microfluidic latch" is used to precisely align a sample after it has been transported a long distance through the flow channels. To demonstrate the scalability of the approach, this paper introduces a "general-purpose" microfluidic chip containing a rotary mixer and addressable storage cells. The system is general purpose in that all operations on the chip operate in terms of unit-sized aqueous samples; using the underlying mechanisms for sample transport and storage, additional sensors and actuators can be integrated in a scalable manner. A novel high-level software library allows users to specify experiments in terms of variables (i.e., fluids) and operations (i.e., mixes) without the need for detailed knowledge about the underlying device architecture. This research represents a first step to provide a programmable interface to the microfluidic realm, with the aim of enabling a new level of scalability and flexibility for lab-on-a-chip experiments. PMID- 16372076 TI - A microfabricated electrical SPLITT system. AB - A growing need for methods to analyze and prepare monodisperse nanoparticles on an industrial scale exists and may be solved by the application of split flow thin fractionation (SPLITT) at the microscale. Microfluidic systems of this type have the ability to separate nanoparticles with high precision in a continuous manner. A miniaturized SPLITT system can be fabricated using standard microfabrication technologies, works in a continuous mode, and can be used as a sample preparation instrument in a micro-total-analysis-system (micro-TAS). In this paper, a miniaturized electrical SPLITT system, which separates particles continuously based on electrophoretic mobility, has been characterized. The advantages of miniaturization have been elucidated. The various aspects of the micro SPLITT system discussed in this paper can be broadly classified into: micro SPLITT system design, fluidics modeling to refine the splitter arrangements, and experimental characterization of the SPLITT system. The design of the micro SPLITT system has been elucidated focusing on the two designs that were implemented. Fluid modeling, used to arrive at a new SPLITT design, was done using a commercially available CFD package to investigate behavior of the fluid in the microchannel with various splitter arrangements. Testing was done with nanoparticles of varying diameter and electrophoretic mobilities to verify the modeling results and demonstrate functionality of the SPLITT system. Particles eluted from both outlets of the SPLITT system were characterized using AFM and SEM to verify the function of the system. PMID- 16372077 TI - Integrated microfluidic UV absorbance detector with attomol-level sensitivity for BSA. AB - An integrated UV absorbance detection system employing a novel silicon-in-plastic technology to seamlessly integrate bare UV photodiode chips into polymer microfluidic systems has been developed. Detection platforms fabricated using this approach exhibit exceptionally low concentration and mass detection limits down to 15 nM and 9.8 amol, respectively, for bovine serum albumin (BSA) as a model protein. In addition to providing high sensitivity, sub-nanoliter detection volumes are enabled by the use of direct photodiode integration. The fabrication methodology is detailed, and system performance metrics including detection limits, detection volume, dynamic range, and linearity are reported. PMID- 16372078 TI - Electronic sorting and recovery of single live cells from microlitre sized samples. AB - Sorting and recovering specific live cells from samples containing less than a few thousand cells have become major hurdles in rare cell exploration such as stem cell research, cell therapy and cell based diagnostics. We describe here a new technology based on a microelectronic chip integrating an array of over 100,000 independent electrodes and sensors which allow individual and parallel single cell manipulation of up to 10,000 cells while maintaining viability and proliferation capabilities. Manipulation is carried out using dynamic dielectrophoretic traps controlled by an electronic interface. We also demonstrate the capabilities of the chip by sorting and recovering individual live fluorescent cells from an unlabeled population. PMID- 16372079 TI - Reflective thermal lens detection device. AB - A reflective thermal lens detection device was developed for realizing a portable and sensitive detector for a microsystem. An aluminum mirror was formed on the main plate of a microchip, and a reflected probe beam was detected with a single pick-up unit. The background signal due to light absorption of the aluminum mirror was 60 times reduced when the microchannel and the mirror were separated with an interval of 600 microm. The tilt angle of the microchip significantly affected the precision of the measurement. Then a quadrant photodiode was used to detect the center of gravity of the reflected probe beam to regulate the tilt angle within +/-0.05 degrees , and this value was enough to achieve 1% CV (coefficient of variance) precision in the measurements. The limit of detection (LOD) was 60 nM for xylene cyanol solution, and the absorbance was 9.4 x 10(-6) AU. About 40 times higher sensitivity was obtained in comparison with a spectrophotometer. PMID- 16372080 TI - Shear force induced monodisperse droplet formation in a microfluidic device by controlling wetting properties. AB - Perpendicular flow is used to induce oil droplet breakup by using a capillary as water phase flow channel. It is a new route to produce monodisperse emulsions. The wetting properties of the fluids on the walls are exceedingly important parameters. Depending on the oil and water flow rates, different spatial distributions of the two phases as laminar, plugs, cobbles and drops, are obtained. The effects of two-phase flow rates on plugs and drop size are studied, and the different droplet formation mechanisms of plug flow and drop flow are discussed. Two quantitative equations utilized to predict the droplet size are developed. PMID- 16372081 TI - Microparticle sampling by electrowetting-actuated droplet sweeping. AB - This paper describes a new microparticle sampler where particles can be efficiently swept from a solid surface and sampled into a liquid medium using moving droplets actuated by the electrowetting principle. We successfully demonstrate that super hydrophilic (2 microm and 7.9 microm diameter glass beads of about 14 degrees contact angle), intermediate hydrophilic (7.5 microm diameter polystyrene beads of about 70 degrees contact angle), and super hydrophobic (7.9 microm diameter Teflon-coated glass beads and 3 microm size PTFE particles of over 110 degrees contact angles) particles on a solid surface are picked up by electrowetting-actuated moving droplets. For the glass beads as well as the polystyrene beads, the sampling efficiencies are over 93%, in particular over 98% for the 7.9 microm glass beads. For the PTFE particles, however, the sampling efficiency is measured at around 70%, relatively lower than that of the glass and polystyrene beads. This is due mainly to the non-uniformity in particle size and the particle hydrophobicity. In this case, the collected particles staying (adsorbing) on the air-to-water interface hinder the droplet from advancing. This particle sampler requires an extremely small amount of liquid volume (about 500 nanoliters) and will thus be highly compatible and easily integrated with lab-on a-chip systems for follow-up biological/chemical analyses. PMID- 16372082 TI - Fabrication of poly(methyl methacrylate) capillary electrophoresis microchips by in situ surface polymerization. AB - A novel method based on in situ surface polymerization of methyl methacrylate (MMA) has been developed for the rapid fabrication of poly(methyl methacrylate) (PMMA) capillary electrophoresis (CE) microchips. MMA containing both thermal and ultraviolet (UV) initiators was allowed to prepolymerize in a water bath to form a fast curing molding solution that was subsequently sandwiched between a nickel template and a PMMA plate. The images of the raised microchannels on the nickel template were precisely replicated into the synthesized PMMA substrates during the UV-initiated polymerization of the molding solution within 30 min under ambient temperature. The attractive performances of the novel PMMA microchips have been demonstrated in connection with amperometric detection for the separation and detection of several model analytes. The new approach significantly simplifies the process for fabricating PMMA devices and could be applied to other materials that undergo light-initiated polymerization. PMID- 16372083 TI - Handheld recirculation system and customized media for microfluidic cell culture. AB - A palm-sized microfluidic recirculation system and customized media enable simplified long-term culture and imaging of cells. The combination of bare Braille display modules, a leveled monolithic surface for complete chip mounting, and a transparent heater improved portability, mechanical stability and optical accessibility. Modification of basal culture media with Leibovitz's L-15 medium enabled an incubator-free culture of carbonate-dependent cells by eliminating the need for exogenous carbon dioxide. This capability is demonstrated through time lapse recording of proliferation of C2C12 myoblasts and MC3T3-E1 osteoblasts for over 2 weeks in ambient atmosphere without medium exchange. The method opens up new possibilities for portable cell culture and for long-term continuous visual monitoring of cells. PMID- 16372084 TI - Organic nonlinear optical materials: where we have been and where we are going. AB - Nonlinear optical (NLO) materials can be useful for a variety of applications varying from modulation of optical signals facilitated by the electro-optic effect-the effect whereby the refractive index of a material changes in response to an applied electric field-to microfabrication, sensing, imaging, and cancer therapy facilitated by multiphoton absorption, wherein molecules simultaneously absorb two or more photons of light. This short Focus article is a brief personal perspective of some of the key advances in second-order NLO materials and in multiphoton-absorbing materials, and of how and why these advances have led to renewed interest in organic NLO materials. PMID- 16372085 TI - Lanthanide-transition metal coordination polymers based on multiple N- and O donor ligands. AB - Lanthanide-transition metal (Ln-M) coordination polymers have attracted extensive interest because they exhibit novel physical properties originating from the interactions between distinct metal ions. This review mainly describes our recent work in the design of Ln-M coordination polymers through the assembly of different metal ions and organic ligands, especially the ligands with multiple N- and O-donor atoms. Many of these crystalline Ln-M materials exhibit intriguing structural motifs and interesting magnetic properties. PMID- 16372086 TI - Quantifying the working stroke of tetrathiafulvalene-based electrochemically driven linear motor-molecules. AB - A highly constrained [2]rotaxane, constructed in such a way that the tetracationic cyclobis(paraquat-p-phenylene) ring is restricted to reside on a monopyrrolotetrathiafulvalene unit, has been synthesised and characterised. This design allows the deslipping free energy barrier for the tetracationic ring in all three redox states of the rotaxane to be determined. PMID- 16372087 TI - High-capacity functional protein encapsulation in nanoengineered polypeptide microcapsules. AB - Addition of polyethylene glycol to aqueous assembly solutions of oppositely charged polypeptides enables high-capacity "loading" of functional protein in biocompatible microcapsules by template-supported layer-by-layer nanoassembly. PMID- 16372088 TI - Probing the B-to-Z-DNA duplex transition using terminally stacking ethynyl pyrene modified adenosine and uridine bases. AB - Pyrene-modified adenosine and uridine bases located in the dangling positions of G,C-alternating oligodeoxynucleotides undergo pi-stacking in their B-DNA duplexes, but not in their Z-DNA duplexes; fluorescence quenching in the former, through photoinduced electron transfer, but not in the latter, allows the state of the B-to-Z-DNA transition to be characterized visually. PMID- 16372089 TI - Synthesis and functionalization of monodisperse poly(ethylene glycol) hydrogel microspheres within polyelectrolyte multilayer microcapsules. AB - Polyelectrolyte multilayer microcapsules were used as templates to prepare monodisperse poly(ethylene glycol) (PEG) hydrogel microspheres, which can react with amine-bearing molecules. PMID- 16372090 TI - Modular assembly of a preorganised, ditopic receptor for dicarboxylates. AB - Two types of calix[4]arene derived hosts for anions with, respectively, 1,3 alternate and cone conformations have been prepared; the 1,3-alternate system binds dicarboxylate anions in a ditopic manner while the cone compounds are deprotonated by carboxylates. PMID- 16372091 TI - Skeletal modification of a non-planar phthalocyanine analogue under very mild conditions. AB - A sterically congested tetraphenyl-substituted dibenzo-tetraaza isobacteriochlorinato nickel derivative unexpectedly reacts with methanol at room temperature to release internal molecular strains, leading to the formation of a ligand having a skeleton bearing two cis-methoxy substituents. PMID- 16372092 TI - Porphyrin-carborane organometallic assemblies based on 1, 2-dicarba-closo dodecaborane (12) ligands. AB - The assembly of soluble, air-stable, supramolecular structures {(Zn TPyP)[Cp*Ir{S2C2(B10H10)}]4(THF)2}(2), {(Cu-TPyP)[Cp*Ir{S2C2(B10H10)}]4(THF)2}(3) and {(Zn-TPyP)[Cp*Ir{S2C2(B10H10)}]2.6(CHCl3)}n (4), based on metal-containing moieties [Cp*Ir{S2C2(B10H10)}] (1) bridged by nitrogen-based organic spacers, are described. PMID- 16372093 TI - Coexistence of spin frustration and long-range magnetic ordering in a triangular Co(II)3(mu3-OH)-based two-dimensional compound. AB - A two-dimensional compound [Co3(mu3-OH)2(1,2-chdc)2]infinity (1,2-chdc = trans 1,2-cyclohexane-dicarboxylate) comprising triangular arrays of Co(II)3(mu3-OH) affording a Kagome-like lattice exhibits the coexistence of spin frustration and long-range magnetic ordering. PMID- 16372094 TI - Pressure induced octahedral tilting distortion in Ba2YTaO6. AB - Herein we communicate the first example of a pressure induced octahedral tilting distortion in a double perovskite phase, which was observed during the structural characterization of Ba2YTaO6 using high-pressure synchrotron X-ray powder diffraction. PMID- 16372095 TI - Sterically non-hindering endocyclic ligands of the bi-isoquinoline family. AB - Bi-isoquinoline can be used as a building block to prepare a new family of non sterically hindering chelates, including a macrocyclic system; the endocyclic nature of the ligands has been confirmed by the X-ray structure of an octahedral tris-chelate iron(II) complex, which shows that the three chelates are easily accommodated in the coordination sphere of the metal in spite of their crescent shape. PMID- 16372096 TI - Crystal structure of an NPNA-repeat motif from the circumsporozoite protein of the malaria parasite Plasmodium falciparum. AB - A crystal structure is reported of the peptide Ac-Ala-Asn-Pro-Asn-Ala-NH2, representing the immunodominant region of the major surface protein on the malaria parasite; the NPNA motif adopts a type-I beta-turn, which is stabilized by hydrogen bonding between the CO of Asn2 and the NH of Ala5 as well as between the O(delta) of Asn2 and the NH of Asn4. PMID- 16372097 TI - Unusual carbon-sulfur bond cleavage in the reaction of a new type of bulky hexathioether with a zerovalent palladium complex. AB - The reaction of a bulky hexathioether, TbtS(o-Phen)S(o-Phen)SS(o-Phen)S(o Phen)STbt (o-Phen = o-phenylene, Tbt = 2,4,6 tris[bis(trimethylsilyl)methyl]phenyl) (1), with 3 molar amounts of Pd(PPh3)4 afforded trinuclear palladium complex bridged by two benzenedithiolato ligands via a three-step palladium insertion reaction into one sulfur-sulfur and two carbon-sulfur bonds of 1. PMID- 16372098 TI - Highly regioselective terminal alkynes hydroformylation and Pauson-Khand reaction catalysed by mesoporous organised zirconium oxide based powders. AB - Zirconia-silica mesoporous powders act as very efficient heterogeneous catalysts for both alkyne hydroformylation and Pauson-Khand reaction and yield regioselectivities opposite to those usually observed. PMID- 16372099 TI - pH-Dependent rectification in self-assembled monolayers based on electrostatic interactions. AB - Asymmetric electrostatic interactions dependent on pH between the redox molecules and the terminal group on the top of the self-assembled monolayer (SAM) afford control of the electron transfer property of the SAM having the imidazole terminal group. PMID- 16372100 TI - Fluorescence modulation in anion sensing by introducing intramolecular H-bonding interactions in host-guest adducts. AB - Fluorescence signaling in anion binding is modulated from quenching to enhancement by intramolecular H-bonding stabilization of anion-ionophore adducts; the intramolecular H-bonding is suggested to suppress the quenching processes otherwise possible and increase the conformational rigidity of the anionic adducts, leading to fluorescence enhancement in a selective fashion towards cyanide ion, among the various anions examined. PMID- 16372101 TI - Core/shell silica-based in-situ microencapsulation: a self-templating method. AB - Core/shell SiO2 and (RSiO1.5)(1-x)-(SiO2)x (R = alkyl) microcapsules were synthesized via a single-step O/W emulsion system using a self-templating method; the facile synthetic process provides an in-situ encapsulation route for a wide range of lipophilic functional compounds. PMID- 16372102 TI - A highly selective synthesis of 3-hydroxy-2-methylpropionamide involving a one pot tandem hydroformylation-hydrogenation sequence. AB - 3-Hydroxy-2-methylpropionamide, an important intermediate in the synthesis of methyl methacrylate, has been obtained with excellent conversion and high selectivity from acrylamide by a tandem hydroformylation-hydrogenation sequence catalysed by Rh/PPh3 and Raney Ni, respectively. PMID- 16372103 TI - Electrochemical annulation of five-membered rings through dearomatization of furans and thiophenes. AB - A new methodology for the annulation of five-membered carbocyclic rings onto enones through the dearomatizing electrochemical cyclization of furans and thiophenes has been developed. PMID- 16372104 TI - Skeletal change in the PNP pincer ligand leads to a highly regioselective alkyne dimerization catalyst. AB - A Rh complex of a bulky diarylamino-based PNP pincer ligand is a robust catalyst for the dimerization of terminal alkynes and highly selective for the trans-enyne product. PMID- 16372105 TI - Determining chemical exchange rates of the uracil labile protons by NMR diffusion experiments. AB - The exchange rates of the amido-protons of uracil with water were determined by NMR diffusion experiments and the results showed a factor 2 difference in lability between them, which was confirmed by more classical 2D-NMR exchange experiments. PMID- 16372106 TI - Flavothionato metal complexes: implications for the use of hydroxyflavothiones as green pesticides. AB - Transition-metal complexes of 3-hydroxyflavothiones have been prepared and structurally characterized; the photochemical properties of these complexes have been examined and are discussed in the context of the use of these compounds as photodegradable pesticides. PMID- 16372107 TI - A novel heterocyclic atom exchange reaction with Lawesson's reagent: a one-pot synthesis of dithiomaltol. AB - A one-pot reaction of maltol with Lawesson's reagent generates dithiomaltol, a thiopyran-4-thione, via an unusual heterocyclic atom exchange (HCAE) reaction; only pyrones with proton or aliphatic substituents undergo the HCAE substitution. PMID- 16372108 TI - Novel highly selective catalytic oxychlorination of phenols. AB - The highly selective oxychlorination of various phenols catalyzed by CuCl2 under mild conditions, in which chloride ions are used as chlorinating agents and dioxygen as a final oxidant, has been developed. PMID- 16372109 TI - Crystal orientation-ordered ZnO nanorod bundles on hexagonal heads of ZnO microcones: epitaxial growth and self-attraction. AB - We demonstrate a preferential nucleation, epitaxial growth, and self-attraction of crystal orientation-ordered ZnO nanorod bundles on (0001) plane of single crystal ZnO microcones. PMID- 16372110 TI - Protonated N'-benzyl-N'-prolyl proline hydrazide as highly enantioselective catalyst for direct asymmetric aldol reaction. AB - Protonated N'-benzyl-N'-l-prolyl-l-proline hydrazide has been developed as a highly enantioselective catalyst for the asymmetric direct aldol reaction of aromatic aldehydes with ketones. PMID- 16372111 TI - No aromaticity of P(6)4- observed via solid state 31P-NMR spectroscopy. AB - The solid state NMR spectra of the binary alkali hexaphosphides Rb4P6 and Cs4P6 unambiguously show the P(6)4- anion not to be aromatic. PMID- 16372112 TI - Layered titanate-zinc oxide nanohybrids with mesoporosity. AB - Zinc oxide-layered titanate nanohybrids with a 1 ratio 1 ordered heterostructure have been successfully synthesized by reassembling exfoliated titanate nanosheets in the sol solution of zinc acetate under hydrothermal conditions. PMID- 16372113 TI - A novel transformation of a zirconium imido compound and the development of a new class of N3 donor heteroscorpionate ligand. AB - Reaction of the dimeric zirconium imido compound [Zr2(mu-NAr)2Cl4(THF)4] with tris(3,5-dimethylpyrazolyl)methyl silane very selectively gave [Zr{(Me2pz)2Si(Me)NAr}Cl3] (1), a highly active pre-catalyst for ethylene polymerisation; a more general and versatile route to N3 donor heteroscorpionate compounds was achieved via the protio ligand (Me2pz)2CHSi(Me)2N(H)iPr for which neutral and cationic organometallic Group 3 and 4 derivatives are reported (Ar = 2,6-C6H(3)iPr2). PMID- 16372114 TI - Recent insights into androgen action on the anatomical and physiological substrate of penile erection. AB - Erectile response is centrally and peripherally regulated by androgens. The original insights into the mechanisms of action of androgens were that androgens particularly exert effects on libido and that erections in response to erotic stimuli were relatively androgen-independent. It was shown that sexual functions in men required androgen levels at the low end of reference values of testosterone. So it seemed that testosterone was not useful treatment for men with erectile difficulties, particularly following the advent of the phosphodiesterase type 5 (PDE5) inhibitors. However, approximately 50% of those treated with PDE5 inhibitors discontinue their treatment. A number of recent developments shed new light on testosterone treatment of erectile dysfunction (ED) in aging men. (1) A recent insight is that, in contrast to younger men, elderly men might require higher levels of testosterone for normal sexual functioning. (2) Several studies have indicated that PDE5 inhibitors are not always sufficient to restore erectile potency in men, and that testosterone improves the therapeutical response to PDE5 inhibitors considerably. (3) There is growing insight that testosterone has profound effects on tissues of the penis involved in the mechanism of erection and that testosterone deficiency impairs the anatomical and physiological substrate of erectile capacity, reversible upon androgen replacement. The synthesis of PDE5 is upregulated by androgens, and the arterial inflow into the penis is improved by giving androgen. The above invites a re-examination of the merits of giving testosterone to aging men with ED. The beneficial effects of PDE5 inhibitors may only be optimally expressed in a eugonadal environment. PMID- 16372115 TI - Sperm chromatin structure and male fertility: biological and clinical aspects. AB - Sperm chromatin/DNA integrity is essential for the accurate transmission of paternal genetic information, and normal sperm chromatin structure is important for sperm fertilizing ability. The routine examination of semen, which includes sperm concentration, motility and morphology, does not identify defects in sperm chromatin structure. The origin of sperm DNA damage and a variety of methods for its assessment are described. Evaluation of sperm DNA damage appears to be a useful tool for assessing male fertility potential both in vivo and in vitro. The possible impact of sperm DNA defects on the offspring is also discussed. PMID- 16372116 TI - Virtual endoscopy of the urinary tract. AB - Technological breakthroughs have advanced the temporal and spatial resolutions of diagnostic imaging, and 3 dimensional (3-D) reconstruction techniques have been introduced into everyday clinical practice. Virtual endoscopy (VE) is a non invasive technique that amplifies the perception of cross-sectional images in the 3-D space, providing precise spatial relationships of pathological regions and their surrounding structures. A variety of computer algorithms can be used to generate 3-D images, taking advantage of the information inherent in either spiral computed tomography or magnetic resonance imaging (MRI). VE images enable endoluminal navigation through hollow organs, thus simulating conventional endoscopy. Several clinical studies have validated the diagnostic utility of virtual cystoscopy, which has high sensitivity and specificity rates in the detection of bladder tumor. Published experience in the virtual exploration of the renal pelvis, ureter and urethra is encouraging but still scarce. VE is a safe, non-invasive method that could be applied in the long-term follow-up of patients with ureteropelvic junction obstruction, urinary bladder tumors and ureteral and/or urethral strictures. Its principal limitations are the inability to provide biopsy tissue specimens for histopathologic examination and the associated ionizing radiation hazards (unless MRI is used). However, in the case of endoluminal stenosis or obstruction, VE permits virtual endoluminal navigation both cephalad and caudal to the stenotic segment. To conclude, VE provides a less invasive method of evaluating the urinary tract, especially for clinicians who are less familiar with cross-sectional imaging than radiologists. PMID- 16372117 TI - Y-chromosomal microdeletions and partial deletions of the Azoospermia Factor c (AZFc) region in normozoospermic, severe oligozoospermic and azoospermic men in Sri Lanka. AB - AIM: To assess for the first time the occurrence of Y chromosomal microdeletions and partial deletions of the Azoospermia Factor c (AZFc) region in Sri Lankan men and to correlate them with clinical parameters. METHODS: In a retrospective study, we analyzed 96 infertile men (78 with non-obstructive azoospermia) and 87 controls with normal spermatogenesis. AZFa, AZFb, AZFc and partial deletions within the AZFc region were analyzed by multiplex polymerase chain reaction (PCR) according to established protocols. RESULTS: No AZFa, AZFb or AZFc deletions were found in the control group. Seven patients in the group of infertile men were found to have deletions as following: one AZFa, two AZFc, two AZFbc and two AZFabc. The relative distribution of these patterns was significantly different compared with that found in the German population. Extension analysis confirmed that the deletions occurred according to the current pathogenic model. gr/gr deletions were found to be equally present both in the patients (n=4) and in the control group (n=4). One b2/b3 deletion was found in the patient group. CONCLUSION: These results suggest that the frequency and pattern of microdeletions of the Y chromosome in Sri Lankan men are similar to those found in other populations and confirm that gr/gr deletions are not sufficient to cause spermatogenetic failure. PMID- 16372118 TI - Application of surface-enhanced laser desorption/ionization time-of-flight-based serum proteomic array technique for the early diagnosis of prostate cancer. AB - AIM: To identify the serum biomarkers of prostate cancer (PCa) by protein chip and bioinformatics. METHODS: Serum samples from 83 PCa patients and 95 healthy men were taken from a mass screening in Changchun, China. Protein profiling was carried out using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). The data of spectra were analyzed using two bioinformatics tools. RESULTS: Eighteen serum differential proteins were identified in the PCa group compared with the control group (P < 0.01). There were four proteins at the higher serum level and 14 proteins at the lower serum level in the PCa group. A decision tree classification algorithm that used an eight-protein mass pattern was developed to correctly classify the samples. A sensitivity of 92.0% and a specificity of 96.7% for the study group were obtained by comparing the PCa and control groups. CONCLUSION: We identified new serum biomarkers of PCa. SELDI-TOF MS coupled with a decision tree classification algorithm will provide a highly accurate and innovative approach for the early diagnosis of PCa. PMID- 16372119 TI - Gene functional research using polyethylenimine-mediated in vivo gene transfection into mouse spermatogenic cells. AB - AIM: To study polyethylenimine (PEI)-mediated in vivo gene transfection into testis cells and preliminary functional research of spermatogenic cell-specific gene NYD-SP12 using this method. METHODS: PEI/DNA complexes were introduced into the seminiferous tubules of mouse testes using intratesticular injection. Transfection efficiency and speciality were analyzed on the third day of transfection with fluorescent microscopy and hematoxylin staining. The long lasting expression of the GFP-NYD-SP12 fusion protein and its subcellular localization in spermatogenic cells at different stages were analyzed with fluorescent microscopy and propidium iodide staining. RESULTS: With the mediation of PEI, the GFP-NYD-SP12 fusion gene was efficiently transferred and expressed in the germ cells (especially in primary spermatocytes). Transfection into Sertoli cells was not observed. The subcellular localization of the GFP-NYD-SP2 fusion protein showed dynamic shifts in spermatogenic cells at different stages during spermatogenesis. CONCLUSION: PEI can efficiently mediate gene transfer into spermatocytes. Thus, it might be useful for the functional research of spermatogenic-cell specific genes such as the NYD-SP12 gene. In our study, the NYD-SP12 protein was visualized and was involved in the formation of acrosome during spermatogenesis. Our research will continue into the detailed function of NYD-SP12 in spermatocytes. PMID- 16372120 TI - A differentially methylated region of the DAZ1 gene in spermatic and somatic cells. AB - AIM: To investigate the methylation status of the deleted in azoospermia 1 (DAZ1) gene promoter region in different cell types. METHODS: Using CpG island Searcher software, a CpG island was found in the promoter region of the DAZ1 gene. The methylation status of this region was analyzed in sperm and leukocytes by bisulfited sequencing. RESULTS: The methylation status of the CpG island in the DAZ1 gene promoter region differed in leukocytes and sperm: it was methylated in leukocytes, but unmethylated in sperm. CONCLUSION: A differentially methylated region of the DAZ1 gene exists in spermatic and somatic cells, suggesting that methylation of this region may regulate DAZ1 gene expression in different tissues. PMID- 16372121 TI - Erectile dysfunction after transurethral prostatectomy for lower urinary tract symptoms: results from a center with over 500 patients. AB - AIM: To identify possible risk factors for erectile dysfunction (ED) after transurethral resection of prostate (TURP) for benign prostatic hyperplasia (BPH). METHODS: Between March 1999 and March 2004, 629 patients underwent TURP in our department for the treatment of symptomatic BPH. All patients underwent transrectal ultrasound examination. In addition, the flow rate, urine residue, International Prostate Symptom Score (IPSS) and quality of life (QOL) were recorded for those who presented without a catheter. Finally, the erectile function of the patient was evaluated according to the International Index of Erectile Function Instrument (IIEF-5) questionnaire. It was determined that ED existed where there was a total score of less than 21. The flow rate, IPSS and QOL assessment were performed at 3 and 6 months post-treatment. The IIEF-5 assessment was repeated at a 6-month follow-up. A logistic regression analysis was used to identify potential risk factors for ED. RESULTS: At baseline, 522 (83%) patients answered the IIEF-5 questionnaire. The mean patient age was (63.7+/-9.7) years. The ED rate was 65%. After 6 months, 459 (88%) out of the 522 patients returned the IIEF questionnaire. The rest of the group was excluded from the statistical analysis. Six months after TURP, the rate of patients reporting ED increased to 77%. Statistical analysis revealed that the only important factors associated with newly reported ED after TURP were diabetes mellitus (P = 0.003, r = 3.67) and observed intraoperative capsular perforation (P = 0.02, r = 1.12). CONCLUSION: The incidence of postoperative, newly reported ED after TURP was 12%. Risk factors for its occurrence were diabetes mellitus and intraoperative capsular perforation. PMID- 16372122 TI - Peyronie's disease: a silent consequence of diabetes mellitus. AB - AIM: To investigate the clinical characteristics of patients with Peyronie's disease (PD) and diabetes mellitus (DM). METHODS: During an 8-year period, a total of 307 men seen at our outpatient clinic were diagnosed with PD. Clinical characteristics, penile deformities and the erectile status of patients with PD and DM together (n=102) were retrospectively analyzed and compared to patients with PD alone with no risk factors for systemic vascular diseases (n=97). RESULTS: The prevalence of PD among men with DM and sexual dysfunction was 10.7%. The mean age of diabetic patients with PD was (55.9+/-8.9) years; in the no risk factor group it was (48.5+/-9.0) years (P < 0.05). The median duration of DM was 5 years. The majority of diabetic patients with PD (56.0%) presented in the chronic phase (P < 0.05), and they were more likely to have a severe penile deformity (>60 degrees) than the no risk factor group (P < 0.05). In the diabetic group, the most common presenting symptom was penile curvature (81.4%), followed by a palpable nodule on the shaft of the penis (22.5%) and penile pain with erection (14.7%). A total of 19.6% of patients were not aware of their penile deformities in the diabetic group. Erectile function, provided by history and in response to intracavernosal injection and a stimulation test, was significantly diminished in patients with PD and DM (P < 0.05). CONCLUSION: DM probably exaggerates the fibrotic process in PD. Diabetic patients with PD have a higher risk of severe deformity and erectile dysfunction (ED). PD seems to be a silent consequence of DM and should be actively sought in diabetic men. PMID- 16372123 TI - Y chromosome microdeletions in azoospermic patients with Klinefelter's syndrome. AB - AIM: To study the occurrence of Y chromosome microdeletions in azoospermic patients with Klinefelter's syndrome (KFS). METHODS: Blood and semen samples were collected from azoospermic patients with KFS (n = 14) and a control group of men of proven fertility (n = 13). Semen analysis was done according to World Health Organization (WHO) guidelines. Blood samples were processed for karyotyping, fluorescent in situ hybridization (FISH) and measurement of plasma follicle stimulating hormone (FSH) by radioimmunoassay. To determine Y chromosome microdeletions, polymerase chain reaction (PCR) of 16 sequence tagged sites (STS) and three genes (DFFRY, XKRY and RBM1Y) was performed on isolated genomic DNA. Testicular fine needle aspiration cytology (FNAC) was done in selected cases. RESULTS: Y chromosome microdeletions spanning the azoospermia factor (AZF)a and AZFb loci were found in four of the 14 azoospermic patients with KFS. Karyotype and FISH analysis revealed that, of the four cases showing Y chromosome microdeletion, three cases had a 47,XXY/46,XY chromosomal pattern and one case had a 46,XY/47,XXY/48,XXXY/48,XXYY chromosomal pattern. The testicular FNAC of one sample with Y chromosome microdeletion revealed Sertoli cell-only type of morphology. However, no Y chromosome microdeletions were observed in any of the 13 fertile men. All patients with KFS had elevated plasma FSH levels. CONCLUSION: Patients with KFS may harbor Y chromosome microdeletions and screening for these should be a part of their diagnostic work-up, particularly in those considering assisted reproductive techniques. PMID- 16372124 TI - Prevalences of oligozoospermia and azoospermia in male partners of infertile couples from different parts of India. AB - AIM: To determine whether there was any regional variation in the prevalence of azoospermia, oligozoospermia and mean sperm counts in male partners of infertile couples from different parts of India. METHODS: Data on 16,714 semen samples analyzed over the past five years from six different laboratories located in five cities of India were collated and evaluated. RESULTS: There was a regional variation in the prevalence of azoospermia. The prevalence of azoospermia was extremely high in Kurnool and Jodhpur (38.3% and 37.4%, respectively). There was also a regional variation in the prevalence of oligozoospermia (51%) in Kurnool. There was no significant difference in the mean sperm counts in normospermic men. CONCLUSION: There is a regional variation in the prevalence of azoospermia and oligozoospermia in the male partners of infertile couples from different regions of India. The prevalence of azoospermia in Kurnool and Jodhpur is higher than any other worldwide reported literature. Further studies need to be carried out to determine the cause of this. PMID- 16372125 TI - Testicular expression of survivin and human telomerase reverse transcriptase (hTERT) associated with spermatogenic function in infertile patients. AB - AIM: To characterize the coexpression of survivin, an inhibitor of apoptosis (IAF), and human telomerase reverse transcriptase (hTERT) in human testes with varying spermatogenic function. METHODS: Transcript levels of survivin mRNA and hTERT mRNA were determined in normal testes (n=11) and testes with defective spermatogenesis (n=28) using real-time reverse-transcription polymerase chain reaction (RT-PCR). The histological work-up was performed according to a modified Johnsen score. RESULTS: Expressions of both survivin and hTERT were highest at median levels of 96.8 and 709 in normal spermatogenesis and dropped to 53.3 and 534 in testes with postmeiotic spermatogenic arrest (n=10). In severe spermatogenic failure (n=18), survivin expression was lacking in most specimens (n=16), whereas at least low levels of testicular hTERT expression were largely detectable with a normalized expression of 73 in premeiotic spermatogenic arrest (n=7) and 45 in patients with Sertoli cell-only syndrome (SCOS) (n=3). Both survivin and hTERT expressions increased with a progressing Johnsen score (P for trend=0.001). CONCLUSION: Although both survivin and hTERT are correlated with spermatogenic function, they show different expression patterns in testes of infertile patients. These findings substantiate results from studies in the rodent testis suggesting a predominant expression of survivin in meiotically dividing germ cells. PMID- 16372126 TI - Benign intratesticular schwannoma: a rare finding. AB - Schwannoma is a peripheral nerve tumour, occasionally located in the genitourinary tract. We described an extremely rare case of intratesticular neurinoma in a 79-year-old patient. PMID- 16372127 TI - Prolonged penile strangulation with metal clamps. AB - Various different objects have been reported to strangulate the penis. We reported on a patient who used metal radiator clamps for an extended period of time. Workup included history, physical examination and urinalysis. The patient was taken to the operating room for further evaluation with cystourethroscopy and orthopedic wire cutters were used to break the metal bands. PMID- 16372128 TI - Left testicular artery arching over the ipsilateral renal vein. AB - AIM: To report two cases of the left testicular artery arching over the left renal vein (LRV) before running downward to the testis. METHODS: The subjects were obtained from two Japanese cadavers. During the student course of gross anatomical dissection, the anatomical relationship between the testicular vessels and the renal vein was specifically observed. RESULTS: The arching left testicular artery arose from the aorta below the LRV and made a loop around the LRV, which appeared to be mildly compressed between the arching artery and the psoas major muscle. CONCLUSION: Clinically, compression of the LRV between the abdominal aorta and the superior mesenteric artery occasionally induces LRV hypertension, resulting in varicocele, orthostatic proteinuria and hematuria. Considering that the incidence of a left arching testicular artery is higher than that of a right one, an arching left artery could be an additional cause of LRV hypertension. PMID- 16372129 TI - Effects of extracts from Hibiscus macranthus and Basella alba mixture on testosterone production in vitro in adult rat testes slices. AB - AIM: To find an in vitro system for the measurement of the androgenic effects of different extracts of Hibiscus macranthus (Malvaceae) and Basella alba (Basellaceae). METHODS: The production of testosterone from testes slices incubated in two media, either Krebs-Henseleit buffer containing 0.5% Bovine serum albumin (BSA) or Dubecco's Modified Eagle's medium-F12 Ham nutrient mixture (DME/Ham F12), under a mixture of 5% CO2 in 95% air was determined either in the presence or absence of cofactors and Hibiscus macranthus plus Basella alba (HMBA) extracts. RESULTS: The testosterone production was increased in testes slices incubated in DME/Ham F12 medium in response to the cofactors (49%) and aqueous extracts (34%-60% according to dilutions). Under the same atmospheric conditions, there was no positive response of the testes slices to either cofactor or HMBA extract stimulation in Krebs-Henseleit buffer containing 0.5% BSA. In further investigations related to the effect of HMBA, the DME/Ham F12 medium was used. The results obtained from the in vitro test showed that the activity was present mainly in methylene chloride and methanol, since these extracts induced an increase in testosterone production by testes slices. CONCLUSION: The testes slice system is suitable to be used for further in vitro investigations of the isolation of androgenic bioactive components of plants. PMID- 16372130 TI - Effect of methanol extract of Ricinus communis seed on reproduction of male rats. AB - AIM: To investigate the effect of methanol extract of Ricinus communis seed (RCE) on male rats reproductive functions. METHODS: Thirty-two male albino rats were divided into four groups. Groups 1, 2 and 3 were gavaged with 0.2 mL of 2.5% tween 80 (RCE vehicle; control) or 20 mg/(kg x d) and 40 mg/(kg x d) of RCE, respectively, for 30 days, and group 4 was also gavaged with 40 mg/(kg x d) of RCE, but was allowed a recovery period of 30 days. Five untreated female rats were cohabited with male rats in each group from day 25 of RCE treatment for 5 days, except group 4, where cohabitation began on day 25 of the recovery period. All male rats were sacrificed 24 h after the experiments. The female rats were laparotomized on day 19 of pregnancy and the number and weight of litters were recorded. RESULTS: There was a significant decrease (P<0.01) in the weight of the reproductive organs, sperm functions and serum levels of testosterone in RCE treated rats. There was disorganization in the cytoarchitecture of the testes, disruption of the seminiferous tubules and erosion of the germinal epithelium. The number and weight of litters of rats in groups 2 and 4 decreased significantly (P<0.05) but no changes were observed in group 3. RCE caused no changes in liver, kidney, heart or body weights in male rats. CONCLUSION: RCE has a reversible negative impact on male reproductive functions, which appears to be mediated via gonadal disruption in testosterone secretion. PMID- 16372131 TI - Endotoxins in the prostatic secretions of chronic prostatitis patients: a need for further biomarkers through the use of proteomics. PMID- 16372132 TI - The CAG repeat at the Huntington disease gene in the Portuguese population: insights into its dynamics and to the origin of the mutation. AB - Huntington disease (HD) is caused by an expansion of a CAG repeat. This repeat is a dynamic mutation that tends to undergo intergenerational instability. We report the analysis of the CAG repeat in a large population sample (2,000 chromosomes) covering all regions of Portugal, and a haplotype study of (CAG)n and (CCG)n repeats in 140 HD Portuguese families. Intermediate class 2 alleles represented 3.0% of the population; and two expanded alleles (36 and 40 repeats, 0.11%) were found. There was no evidence for geographical clustering of the intermediate or expanded alleles. The Portuguese families showed three different HD founder haplotypes associated with 7-, 9- or 10-CCG repeats, suggesting the possibility of different origins for the HD mutation among this population. The haplotype carrying the 7-CCG repeat was the most frequent, both in normal and in expanded alleles. In general, we propose that three mechanisms, occurring at different times, may lead to the evolution from normal CAGs to full expansion: first, a mutation bias towards larger alleles; then, a stepwise process that could explain the CAG distributions observed in the more recent haplotypes; and, finally, a pool of intermediate (class 2) alleles more prone to give rise to expanded HD alleles. PMID- 16372133 TI - Comparison of the effects of agalsidase alfa and agalsidase beta on cultured human Fabry fibroblasts and Fabry mice. AB - We compared two recombinant alpha-galactosidases developed for enzyme replacement therapy for Fabry disease, agalsidase alfa and agalsidase beta, as to specific alpha-galactosidase activity, stability in plasma, mannose 6-phosphate (M6P) residue content, and effects on cultured human Fabry fibroblasts and Fabry mice. The specific enzyme activities of agalsidase alfa and agalsidase beta were 1.70 and 3.24 mmol h(-1) mg protein(-1), respectively, and there was no difference in stability in plasma between them. The M6P content of agalsidase beta (3.6 mol/mol protein) was higher than that of agalsidase alfa (1.3 mol/mol protein). The administration of both enzymes resulted in marked increases in alpha galactosidase activity in cultured human Fabry fibroblasts, and Fabry mouse kidneys, heart, spleen and liver. However, the increase in enzyme activity in cultured fibroblasts, kidneys, heart and spleen was higher when agalsidase beta was used. An immunocytochemical analysis revealed that the incorporated recombinant enzyme degraded the globotriaosyl ceramide accumulated in cultured Fabry fibroblasts in a dose-dependent manner, with the effect being maintained for at least 7 days. Repeated administration of agalsidase beta apparently decreased the number of accumulated lamellar inclusion bodies in renal tubular cells of Fabry mice. PMID- 16372134 TI - Quantitative trait loci in ABCA1 modify cerebrospinal fluid amyloid-beta 1-42 and plasma apolipoprotein levels. AB - The ATP-binding cassette transporter A1 encoded by ABCA1 plays an integral role in the efflux of cellular cholesterol and phospholipids, but may also be a central mediator of beta-amyloid (Abeta) processing. Here, genetic association of the common R219K variant of ABCA1 is shown with cerebrospinal fluid (CSF) Abeta 1 42 levels, reinforcing emerging evidence of a connection between lipid and Abeta metabolism. In support of this finding we demonstrate for the first time that CSF cholesterol and Abeta 1-42 are correlated. To affirm the plausible impact of ABCA1 variation on cholesterol and related traits as well as to empower a survey of possible interactions (e.g. age, gender, and smoking), a large Swedish population consisting of over 2,700 individuals was enlisted and extensive measures of plasma lipid parameters carried out. These analyses revealed that R219K has a strong effect on apolipoprotein B (APOB) and LDL-cholesterol (LDL-C) among smokers (P = 0.000055 and P = 0.00059, respectively), but not among non smokers. In contrast, no effect was evident with apolipoprotein A (APOA1) or HDL cholesterol (HDL-C) levels. Plasma APOB and LDL-C, but not APOA1 and HDL-C, were shown to be markedly elevated in smokers versus non-smokers, affirming that smoking may selectively impact the former pathway. No other genetic markers in ABCA1 exhibit effects as large as R219K, although a modest independent effect of R1587K was observed. Our data illuminate a possible genetic link between Abeta and cholesterol metabolism, but also provide an intriguing example of an environmental exposure that may modify a genotype-phenotype relationship. PMID- 16372135 TI - Mutation analysis of the GNE gene in Korean patients with distal myopathy with rimmed vacuoles. AB - Distal myopathy with rimmed vacuoles (DMRV; MIM 605820) is an autosomal recessive neuromuscular disorder characterized by weakness of the anterior compartment of the lower limbs, sparing the quadriceps muscles. Recently, mutations in the UDP-N acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE) gene have been identified as the genetic basis of DMRV. To investigate the mutation spectrum of the GNE gene in Korean patients with DMRV, we performed clinical and genetic analysis of nine unrelated patients suspected to have DMRV. Direct sequencing analysis revealed that eight out of nine patients (88.9%) were either homozygous or compound heterozygous for GNE gene mutations, including three known (C13S, R129Q, and V572L) and two novel mutations (M29T and A591T) [corrected] The allelic frequencies of the V572L and C13S mutations were 68.8% (11/16) and 12.5% (2/16), respectively. These results suggest that screening for GNE gene mutations in patients suspected to have DMRV would be helpful for molecular diagnosis of DMRV in the Korean population. PMID- 16372136 TI - Association study of semaphorin 7a (sema7a) polymorphisms with bone mineral density and fracture risk in postmenopausal Korean women. AB - Bone mineral density (BMD), the major factor determining bone strength, is closely related to osteoporotic fracture risk and is determined largely by multiple genetic factors. Semaphorin 7a (SEMA7A), a recently described member of the semaphorin family, has been shown to play a critical role in the activation of monocyte/macrophages that share progenitors with bone-resorbing osteoclasts and thus might contribute to osteoclast development. In the present study, we directly sequenced the SEMA7A gene in 24 Korean individuals, and identified 15 sequence variants. Five polymorphisms (+15667G > A, +15775C > G, +16285C > T, +19317C > T, +22331A > G) were selected and genotyped in postmenopausal Korean women (n = 560) together with measurement of the areal BMD (g/cm2) of the anterior-posterior lumbar spine and the non-dominant proximal femur using dual energy X-ray absorptiometry. We found that polymorphisms of the SEMA7A gene were associated with the BMD of the lumbar spine and femoral neck. SEMA7A + 15775C > G and SEMA7A+22331A > G were associated with low BMD of the femoral neck (P = 0.02) and lumbar spine (P = 0.04) in a recessive model. SEMA7A-ht4 also showed an association with risk of vertebral fracture (OR = 1.87-1.93, P = 0.02-0.03). Our results suggest that variations in SEMA7A may play a role in decreased BMD and risk of vertebral fracture. PMID- 16372137 TI - A novel Wiskott-Aldrich syndrome protein (WASP) complex mutation identified in a WAS patient results in an aberrant product at the C-terminus from two transcripts with unusual polyA signals. AB - Wiskott-Aldrich syndrome (WAS) is an X-linked recessive disorder characterized by immunodeficiency, thrombocytopenia and eczema. A broad spectrum of mutations in the WASP gene has been identified as causing the disease. In the present paper, we report on a patient affected by WAS with a novel complex mutation, characterized by a small 9 bp deletion followed by an inversion of 151 bp and a gross deletion of 4.3 kb within the Xp11.23 region. The small deletion and the inverted fragment are found in intron 11. The large deletion initiates downstream of exon 11 of the WASP gene, including exon 12, and a genomic region upstream of the promoter of the contiguous SUV39H1 gene. Expression studies of the mRNA of the patient's sample showed the presence of two aberrant transcripts that code for a protein of 519 amino acids. We demonstrate that these two transcripts differ in the 3' UTR region, and result from the use of two alternative polyadenylation signals. The severe phenotype of the patient correlates with the presence of an aberrant protein. PMID- 16372138 TI - [Biologics for treatment of juvenile idiopathic arthritis. Consensus statement of the 7th Worlitzer Expertengesprache 2004 for the German Arbeitsgemeinschaft Kinder- und Jugendrheumatologie]. AB - The group of biologics for the treatment of rheumatic diseases is continuously growing. They have become an important option not only for treatment of so far untreatable chronic inflammatory or rheumatic disease, but also for juvenile idiopathic arthritis. In addition, the velocity and the degree of improvement is better than with to conventional therapies. Furthermore, toxicity and risks seem to be lower with higher safety and compatibility. Although the data are scarce, they are widely used. Therefore, the German Arbeitsgemeinschaft Kinder- und Jugendrheumatologie is updating the current recommendation for the treatment of juvenile idiopathic arthritis using biologics. PMID- 16372140 TI - Single coronary artery presenting with cardiogenic shock due to acute myocardial infarction. AB - Percutaneous coronary revascularization plays an important role in the management of acute coronary syndrome. Unpredictable angiographic findings of anomalous coronary arteries may, however, compromise the otherwise high and predictable success rates of this intervention. We report a case of failed coronary angioplasty of the left anterior descending artery through an anomalous left main coronary artery originating from the right coronary sinus in a 33-year-old man with acute myocardial infarction complicated by cardiogenic shock. Subsequently the patient performed successful emergency coronary artery bypass graft. PMID- 16372141 TI - Percutaneous myocardial ablation for left mid-ventricular obstructive hypertrophic cardiomyopathy. AB - Mid-ventricular obstructive hypertrophic cardiomyopathy (MVOHC) is a rare type of cardiomyopathy. The diagnosis is based on the hourglass appearance on the left ventriculogram and the presence of pressure gradient between apical and basal chamber of the ventriculum on the hemodynamic assessment. Atrio-ventricular sequential pacing and alcohol ablation have been proposed as less invasive alternatives to surgery for patients who fail to respond to pharmacologic therapy. The present case represents successful percutaneous treatment with alcohol ablation to patient with MVOHC and apical aneurysm. Alcohol ablation to distal segment of the left anterior descending artery resulted with relief of clinical symptoms and intra-ventricular pressure gradient. PMID- 16372139 TI - Imaging techniques in nuclear cardiology for the assessment of myocardial viability. AB - The assessment of myocardial viability has become an important aspect of the diagnostic and prognostic work-up of patients with ischemic cardiomyopathy. Although revascularization may be considered in patients with sufficient viable myocardium, patients with predominantly scar tissue should be treated medically. Patients with left ventricular dysfunction who have viable myocardium are the patients at highest risk because of the potential for ischemia but at the same time benefit most from revascularization. It is important to identify viable myocardium in these patients, and radionuclide myocardial scintigraphy is an excellent tool for this. Single-photon emission computed tomography perfusion scintigraphy (SPECT), whether using (201)thallium, (99m)Tc-sestamibi, or (99m)Tc tetrofosmin, in stress and/or rest protocols, has consistently been shown to be an effective modality for identifying myocardial viability and guiding appropriate management. Metabolic and perfusion imaging with positron emission tomography (PET) radiotracers frequently adds additional information and is a powerful tool for predicting which patients will have an improved outcome from revascularization. New techniques in the nuclear cardiology field, like attenuation corrected SPECT, dual isotope simultaneous acquisition (DISA) SPECT and gated FDG PET are promising and will further improve the detection of myocardial viability. Also the combination of multislice computed tomography scanners with PET opens possibilities of adding coronary calcium scoring and non invasive coronary angiography to myocardial perfusion imaging and quantification. Evaluation of the clinical role of these creative new possibilities warrants investigation. PMID- 16372142 TI - Suicidal behaviour in national and international adult adoptees: a Swedish cohort study. AB - BACKGROUND: Previous studies have shown an elevated risk for suicidal behaviour in adolescent and young adult international adoptees. Comparisons between national and international adoptees in this respect have been inconclusive. METHODS: A total of 6,065 international adoptees were compared to 7,340 national adoptees and 1,274,312 non-adopted study subjects, all born between 1963 and 1973 and followed up until 2002 using the National Swedish Registers. Cox regression of person years was used in multivariate analyses to compare risks for suicide death and suicide attempt. RESULTS: International adoptees had clearly increased risks for suicide attempt (risk ratio 4.5 [95% confidence interval 3.7-5.5]) and suicide death (3.6 [2.6-5.2]) after adjustments for sex, age and socio-economic factors. National adoptees had lower risks than international adoptees, but had increased risks compared to non-adoptees (suicide attempt, 2.8 [2.2-3.5]; suicide death, 2.5 [1.8-3.3]). Biological parents' morbidity explained approximately one third of the increased risk for national adoptees. Female international adoptees' risk for suicide attempt was elevated to an even greater extent than in male international adoptees, when compared to the general population. CONCLUSIONS: Clinicians should be aware that an increased risk for suicide and suicide attempts in international adoptees is a topic that is equally relevant to child and adult psychiatry. PMID- 16372143 TI - Helping alliance and early dropout from psychiatric out-patient care: the influence of patient factors. AB - BACKGROUND: The study examined client factors of relevance in the establishment of helping alliance and in the prediction of dropout from a routine psychiatric setting admitting a variety of diagnoses and staffed with a multiprofessional team. METHOD: Newly admitted patients (n=122) and staff completed questionnaires regarding helping alliance, and the patients also completed questionnaires regarding motivation, symptoms and interpersonal problems. The patients were also diagnosed according to ICD-10 and were followed up concerning early dropout. RESULTS: Several variables correlated with helping alliance, and multivariate analyses showed that cold/distant factor, motivation and interpersonal sensitivity factor were the most important factors in establishing helping alliance. Moreover, it was the alliance as perceived by the patients (and not by the staff) that proved to be the most essential variable. A logistic regression analysis showed that early dropout was predicted by low helping alliance, low age and cold/distant factor. CONCLUSION: The most important client factors for establishing helping alliance and for predicting early dropout seem to be those relevant to interpersonal processes. Furthermore, the therapists'/staff's responsiveness to these client factors seems to be of decisive importance. PMID- 16372144 TI - Midazolam inhibits neophobia-induced Fos expression in the rat hippocampus. AB - The effect of midazolam on expression of c-Fos protein was examined in the rat hippocampus, following the open field test of neophobia. It was found that pretreatment of rats with midazolam, at the dose of 0.5 mg/kg, enhanced rat exploratory behavior, and inhibited neophobia related stimulation of c-Fos in the CA-1 and CA-3 areas of the hippocampus. The presented results provide new immunocytochemical data on the involvement of hippocampus in emotional processes related to neophobia, and indicate a possible site of action of benzodiazepines. PMID- 16372145 TI - The effects of exogenous melatonin on the morphology of thyrocytes in pinealectomized and irradiated rats. AB - We investigated the effects of exogenous melatonin on the thyrocytes morphology in gamma-irradiated rats under condition where the pineal gland, as a main physiological source of endogenous melatonin, was removed. Three months after pinealectomy animals were divided into two groups: one group of animals was treated with 0.5 ml of vehicle (ethanol diluted in water) and other group was injected intraperitoneally 2 mg/kg of melatonin dissolved in 0.5 ml of vehicle daily during the period of fourteen days. After this treatment all the animals were irradiated with a single dose of 8 Gy gamma rays. Ionising radiation induced apoptosis, hydropic swelling or/and necrosis in both groups of animals, however these changes were less discerned in the thyrocytes of melatonin-treated animals. Our findings demonstrate that administration of exogenous melatonin prior to irradiation reduces radiation-induced thyrocytes damage. PMID- 16372146 TI - Lisuride treatment of restless legs syndrome: first studies with monotherapy in de novo patients and in combination with levodopa in advanced disease. AB - In two 4-week polysomnography pilot studies with 10 patients each, we investigated the efficacy of oral lisuride as monotherapy in de novo RLS patients as well as in combination with levodopa in advanced RLS. Daily doses at study end were 0.3 mg lisuride, plus 150 mg levodopa in the combination study. Marked improvements occurred in both studies in different PLM indexes and in the CGI. Levodopa dose could be decreased by 27%. Lisuride might be an efficacious treatment for RLS in general, and in combination with levodopa in advanced stage. PMID- 16372147 TI - Ultrasonography and fine-needle aspiration cytology can spare breast cancer patients unnecessary sentinel lymph node biopsy. AB - BACKGROUND: Some 30% to 40% of the breast cancer patients scheduled for sentinel node biopsy have axillary metastasis. Pilot studies suggest that ultrasonography is useful in the preoperative detection of such nodes. The aims of this study were to evaluate the sensitivity of preoperative ultrasonography and fine-needle aspiration cytology for detecting axillary metastases and to assess how often sentinel node biopsy could be avoided. METHODS: Between October 1999 and December 2003, 726 patients with clinically negative lymph nodes were eligible for sentinel node biopsy. A total of 732 axillae were examined. Preoperative ultrasonography with subsequent fine-needle aspiration cytology in case of suspicious lymph nodes was performed in all patients. The sentinel node procedure was omitted in patients with tumor-positive axillary lymph nodes in lieu of axillary lymph node dissection. RESULTS: Ultrasound and fine-needle aspiration cytology established axillary metastases in 58 (8%) of the 726 patients. These 58 were 21% of the total of 271 patients who were proven to have axillary metastasis in the end. Of the patients with ultrasonographically suspicious lymph nodes and negative cytology, 31% had tumor-positive sentinel nodes. Patients with preoperatively established metastases by ultrasonography and fine-needle aspiration cytology had more tumor-positive lymph nodes (P < .001) than patients with metastases established later on. CONCLUSIONS: The sensitivity of ultrasonography and fine-needle aspiration cytology is 21%, and unnecessary sentinel node biopsy is avoided in 8% of the patients. This approach improves the selection of patients eligible for sentinel node biopsy. PMID- 16372148 TI - Predicting extensive nodal disease in women with breast cancer. PMID- 16372149 TI - Does prior adjuvant chemoradiotherapy lead to a safer pancreatoduodenectomy? PMID- 16372150 TI - Decision making for patients with colorectal cancer liver metastases. PMID- 16372151 TI - Unresectable hepatic colorectal metastases: need for new surgical strategies. PMID- 16372152 TI - Method of presenting oncology treatment outcomes influences patient treatment decision-making in metastatic colorectal cancer. AB - BACKGROUND: The methods used to communicate relevant outcomes in oncology to patients will likely influence treatment decisions. The purpose of this study was to examine the influence of three different methods of describing the efficacy of therapy on treatment decisions regarding management of metastatic colorectal cancer. METHODS: Participants reviewed a clinical scenario and randomly received one of three ways of describing efficacy of chemotherapy in metastatic colorectal cancer: (1) relative risk reduction, (2) tumor response rate, and (3) median overall survival. They received the same clinical scenario but were presented four treatment options: (1) observation and supportive care, (2) chemotherapy, (3) surgery, and (4) surgery and chemotherapy and the accompanying median overall survival estimate. RESULTS: Participants included 102 preclinical medical students. In the first scenario, 85% chose chemotherapy in the relative risk reduction group, as did 88% of the tumor response rate group, but significantly fewer participants did so in the median overall survival group (35%; P < .001). In the second scenario, there was a significant difference in treatment preferences, with 4% of participants choosing observation/supportive care. None chose chemotherapy only, 19% chose surgery only, and 77% chose surgery plus chemotherapy (P < .001). CONCLUSIONS: This study demonstrated that different methods of describing oncology treatment outcomes associated with therapy for metastatic colorectal cancer to the liver can have a dramatic effect on patient treatment decisions. PMID- 16372153 TI - Inhibition of host signal transducer and activator of transcription factor 6 results in cure with cyclophosphamide and interleukin 12 immunotherapy. AB - BACKGROUND: Interleukin (IL)-12 immunotherapy is highly effective against established immunogenic tumors. However, nonimmunogenic tumors fail to respond to IL-12 therapy. Analysis of tumor rejection of the immunogenic tumors shows that a preexisting antitumor immune response is required for an effective IL-12 response. It is not known whether this lack of a preexisting host antitumor immune response is a limiting factor for the lack of response to IL-12 therapy by nonimmunogenic tumors. METHODS: Experiments were done using the spontaneously arising nonimmunogenic metastatic murine breast 4T1 carcinoma in normal and STAT6 knockout BALB/c mice. RESULTS: 4T1 is nonimmunogenic in normal mice, and established subcutaneous tumors are resistant to immunotherapy with cyclophosphamide (Cy) plus IL-12. However, in STAT6 knockout mice, 4T1 becomes immunogenic, and established 4T1 tumors are eradicated by Cy plus IL-12. Adoptive transfer of spleen cells from normal mice into STAT6 knockout mice before tumor inoculation reduces both the immunogenicity and response to Cy plus IL-12 immunotherapy of 4T1 in the recipient mice. CONCLUSIONS: Cy plus IL-12 immunotherapy can eradicate nonimmunogenic tumors as long as a preexisting immunity is established in the tumor-bearing host. Furthermore, the STAT6 pathway is likely involved in the suppression of the development of host antitumor immunity. PMID- 16372154 TI - Effect of neoadjuvant chemoradiation on operative mortality and morbidity for pancreaticoduodenectomy. AB - BACKGROUND: Neoadjuvant chemoradiotherapy (neo-CRT) is being used with increasing frequency for periampullary tumors, but how it alters the complication rate of pancreaticoduodenectomy (PD) is unclear. METHODS: A retrospective analysis was conducted of 79 patients with periampullary malignancies who received 5 fluorouracil-based neo-CRT followed by PD. RESULTS: There was no difference in mortality between PD after neo-CRT (3.8%) and conventional PD for either malignant (4.5%) or benign (2.2%) disease. Focusing only on patients with malignancy, the neo-CRT group had a significantly lower pancreatic leak rate than the conventional group (10% vs. 43%; P < .001). Intra-abdominal abscesses were less common in the neo-CRT group (8.8% vs. 21%; P = .019), and there was one (1.2%) amylase-rich abscess in neo-CRT group, compared with eight (12%) in the conventional group. In addition, two patients in the conventional group died of leak-associated sepsis, compared with none in the neo-CRT group. Multivariate analysis revealed that neoadjuvant chemoradiation (odds ratio, .15) was the most significant factor associated with a reduced risk of pancreatic leak. CONCLUSIONS: Neo-CRT does not increase the mortality or morbidity of PD. In contrast, neo-CRT was associated with a marked reduction in the incidence of pancreatic leak, as well as leak-associated morbidity and mortality. PMID- 16372155 TI - Lymphatic mapping and sentinel lymph node biopsy for patients with local recurrence after breast-conservation therapy. AB - BACKGROUND: Local recurrence (LR) after breast-conservation therapy for breast cancer occurs in 10% to 15% of cases. A subset of these represents biologically aggressive disease, yet prognostic features for identifying this high-risk category are lacking. We hypothesized that lymphatic mapping and sentinel lymph node biopsy would provide useful information regarding dominant lymphatic drainage patterns of patients with LR. METHODS: Breast cancer case records involving surgery for LR at the University of Michigan from 2002 to 2004 were reviewed. The lymphatic drainage patterns were compared with those of 117 patients who underwent mapping for primary breast cancer. RESULTS: Fourteen LR cases were identified (10 with initial axillary lymph node dissection, 2 with initial sentinel lymph nodes, and 2 with no axillary surgery at the time of primary cancer treatment); lymphatic mapping was performed in 10. The sentinel lymph node identification rate was 90%, the median number of lymph nodes retrieved was 3, and no metastases were detected. Significantly more cases of nonipsilateral axillary sentinel node drainage were observed in mapping procedures performed for LR compared with those for primary breast cancer (67% vs. 15%; P = .001). CONCLUSIONS: Lymphatic mapping is feasible in patients undergoing mastectomy for LR and is likely to identify aberrantly located sentinel lymph nodes that would otherwise be overlooked with a conventional completion mastectomy. PMID- 16372156 TI - Prognostic effect of re-excision in adult soft tissue sarcoma of the extremity. AB - BACKGROUND: We explored the outcome of patients with primary adult soft tissue sarcoma (STS) of the extremities undergoing re-excision after previous unplanned surgery. METHODS: A total of 597 consecutive adult patients with primary extremity STS were treated with conservative surgery at our institution over a 20 year time span. A total of 318 patients were referred after unplanned excisions, and the remaining 279 underwent primary resection at our center. The two groups significantly differed in tumor size and depth. The assessed end points were sarcoma-specific mortality, local recurrence, and distant metastasis. Univariable and multivariable analyses, adjusted for other prognostic factors, were performed in the competing risks framework. RESULTS: The adjusted 10-year cumulative incidences in re-excised and primarily operated patients were, respectively, 18.7% and 16.4% (P = .535) for local relapse, 17.6% and 20.2% (P = .541) for metastasis, and 20.4% and 22.4% (P = .645) for mortality. Among patients who underwent re-excision, evidence of microscopic residual disease on pathologic examination had a significant prognostic effect on multivariable analysis for distant metastases (P = .002). A trend for survival was detected as well. CONCLUSIONS: At a referral center with a liberal policy of re-excisions in adult primary STS of the extremities, the outcome of patients who underwent re-excision was similar to that of patients who had primary resections. Evidence of microscopic residual disease at re-excision was a marker of clinical aggressiveness. PMID- 16372157 TI - Solitary and multiple isolated metastases of clear cell renal carcinoma to the pancreas: an indication for pancreatic surgery. AB - BACKGROUND: Isolated pancreatic metastases (isPMs) of clear cell renal carcinoma are rare. Most of them are solitary; some are multiple. The reported rates and the clinical implications of multiple isPMs from clear cell renal cancer vary. Therefore, the available literature was analyzed to shed light on the clinical significance of these extremely rare metastatic lesions. METHODS: A literature search brought to light 236 cases of isPMs (both solitary and multiple) from renal cell carcinoma. These were analyzed. RESULTS: A total of 12% of the metastases were synchronous with the primary tumor, and 88% were metachronous, occurring 10.0 +/- 6.5 years (mean +/- SD) after nephrectomy. A predilection for a specific part of the pancreas was not identifiable. The localization of the renal cell cancer (left or right kidney) did not have any effect on the site of the metastases. Seventy-four (39%) of the metastases to the pancreas were multiple (3.2 +/- 1.5). Their epidemiology did not differ from that of solitary metastatic lesions. Actuarial 3- and 5-year survival rates after radical resection were 78% and 78%, respectively, for multiple versus 75% and 64% for solitary metastases. CONCLUSIONS: The epidemiological data do not support a direct local lymphogenous or venous spread from the primary tumor to the pancreas. They rather suggest a systemic spread. Because of the positive outcome, radical removal of both solitary and multiple metastases should be attempted in eligible patients. PMID- 16372158 TI - Treatment with 5-fluorouracil/folinic acid, oxaliplatin, and irinotecan enables surgical resection of metastases in patients with initially unresectable metastatic colorectal cancer. AB - BACKGROUND: The prognosis of unresectable metastatic colorectal cancer might be improved if a radical surgical resection of metastases could be performed after a response to chemotherapy. METHODS: We treated 74 patients with unresectable metastatic colorectal cancer (not selected for a neoadjuvant approach) with irinotecan, oxaliplatin, and 5-fluorouracil/leucovorin (FOLFOXIRI and simplified FOLFOXIRI). Because of the high activity of these regimens (response rate, 72%), a secondary curative operation could be performed in 19 patients (26%). RESULTS: Four patients underwent an extended hepatectomy, nine patients underwent a right hepatectomy, three patients underwent a left hepatectomy, and three patients had a segmental resection. In five patients, surgical removal of extrahepatic disease was also performed. In seven patients, surgical resection was combined with intraoperative radiofrequency ablation. The median overall survival of the 19 patients who underwent operation is 36.8 months, and the 4-year survival rate is 37%. The median overall survival of the 34 patients who were responsive to chemotherapy, but who did not undergo operation, is 22.2 months (P = .0114). CONCLUSIONS: The FOLFOXIRI regimens we studied have significant antitumor activity and allow a radical surgical resection of metastases in patients with initially unresectable metastatic colorectal cancer not selected for a neoadjuvant approach and also those with extrahepatic disease. The median survival of patients with resected disease is promising. PMID- 16372159 TI - Production of eicosapentaenoic and docosahexaenoic acid-containing oils in transgenic land plants for human and aquaculture nutrition. AB - A large body of evidence suggests that there is a significant underconsumption of omega-3, long-chain, polyunsaturated fatty acids (LC-PUFAs) and that this is the cause of multiple chronic diseases and developmental aberrations. The scope for increasing omega-3 LC-PUFA consumption from seafood is limited because global wild fisheries are unable to increase their harvests, and aquaculture fisheries currently rely on wild fisheries as a source of LC-PUFAs. Agricultural production of oils is highly efficient and has the potential to be sustainable. The transfer of genes from marine microalgae and other microorganisms into oilseed crops has shown that the production of terrestrial omega-3 LC-PUFA oils is indeed possible. The specifications of these oils or whole seeds for use in human and Atlantic salmon (Salmo salar) aquaculture nutrition are discussed. PMID- 16372160 TI - Isolation, characterization, and expression analysis of three actin genes in the New Zealand black-footed abalone, Haliotis iris. AB - Three actin genes -- H. irisA1, H. irisA2, and H. irisA3 -- from the mollusc Haliotis iris (New Zealand black-footed abalone) were isolated by polymerase chain reaction (PCR). The genes were similar to molluscan (84.1% to 94.9%) and vertebrate (84.5% to 86.6%) actins. The sequence similarity between the genes ranged from 88.5% to 93.2%. The greatest disparity, 32.3%, was found over a 99-nt region located at nt 808-906 of H. irisA1, corresponding to amino acids 212-244 of the three actins. The H. iris actin gene family contains at least eight members. Reverse transcription (RT)-PCR analysis of the three genes showed H. irisA1 and H. irisA2 were expressed at low levels in fertilized eggs and blastula stages and at high levels in trochophore and veliger larvae. H. irisA3 was detected in fertilized eggs; it was not detected in the blastula stages and at high levels in the trochophore and veliger larvae. The structure and expression of the three actin genes are discussed. PMID- 16372162 TI - Global left ventricular function in cardiac CT. Evaluation of an automated 3D region-growing segmentation algorithm. AB - The purpose was to evaluate a new semi-automated 3D region-growing segmentation algorithm for functional analysis of the left ventricle in multislice CT (MSCT) of the heart. Twenty patients underwent contrast-enhanced MSCT of the heart (collimation 16 x 0.75 mm; 120 kV; 550 mAseff). Multiphase image reconstructions with 1-mm axial slices and 8-mm short-axis slices were performed. Left ventricular volume measurements (end-diastolic volume, end-systolic volume, ejection fraction and stroke volume) from manually drawn endocardial contours in the short axis slices were compared to semi-automated region-growing segmentation of the left ventricle from the 1-mm axial slices. The post-processing-time for both methods was recorded. Applying the new region-growing algorithm in 13/20 patients (65%), proper segmentation of the left ventricle was feasible. In these patients, the signal-to-noise ratio was higher than in the remaining patients (3.2+/-1.0 vs. 2.6+/-0.6). Volume measurements of both segmentation algorithms showed an excellent correlation (all P 1,500 g. In regard to gestation, ROP was found in 27 (86.2%) babies born at < 27 weeks of gestation, in 87 (35.6%) born at 27-30 weeks of gestation and in 19 infants (12.7%) born at > 30 weeks gestation. There were 75 babies with stage III disease and 73.3% needed treatment. Retinal ablation was performed at 62.9 days. CONCLUSIONS: ROP incidence is inversely related to birth weight and gestational age. Three out of four babies who reached stage III disease needed treatment. The effectiveness of photocoagulation was over 90% when performed at 2 months of age. PMID- 16372215 TI - [Spatial summation: its topography in the central visual field]. AB - PURPOSE: Luminance (L) and area (A) of the stimulus are related by the equation LxAk=constant. We evaluated the k value in 66 positions of the central visual field. METHODS: Ten eyes of 10 healthy subjects were examined for conventional luminous thresholds at 66 positions of the central visual field with the TOP strategy using Goldmann sizes 4, 3.5, 3, 2.5 and 1.9 in the PULSAR perimeter. RESULTS: The k value increased in a lineal manner from the centre towards the periphery (slope=0.01 per degree, average k=0.616, r=0.98, p<0.01) but with unequal slopes at the different meridians. In the inferior hemi-fields it was higher (k=0.657) than in the superior hemi-fields (k=0.574). The k value at the supero-nasal hemifields is quite similar to that found at the paracentral region, and maximum at the temporal inferior region. CONCLUSIONS: Spatial summation in the central visual field has specific values for different positions. PMID- 16372216 TI - [Ocular infiltration in a patient with Multiple Myeloma]. AB - CASE REPORT: We report the case of a patient with Multiple Myeloma Ig G Kappa stage IIIA and hepatic infiltration, who presented with ocular infiltration. The initial sign was a rapidly growing sub-conjunctival mass that caused ophthalmoplegia and increased intraocular pressure secondary to progressive infiltration of the orbit. DISCUSSION: Multiple Myeloma is a malignant tumor of plasma cells that can affect the eye and orbit, not only as an infiltration secondary to the disease but also primary as an isolated tissue proliferation (extramedullary plasmacytoma). Chemotherapy is the usual treatment given. Radiotherapy with or without excisional surgery is the first option for plasmacytomas. PMID- 16372217 TI - [Treatment with topical interferon alone in severe conjunctivo-corneal neoplasia]. AB - CASE REPORT: A 78-year-old man was referred for evaluation of a limbic lesion in his left eye. This had a gelatinous appearance and an extension of 240 degrees . The lesion was clinically diagnosed to be a conjunctival and corneal intraepithelial neoplasia (CIN). Topical treatment with interferon alpha2b (INFalpha2b) four times daily was started and continued until the lesion had completely resolved at 4 months. DISCUSSION: Topical INFalpha2b alone may be a safe and effective treatment of CIN instead of the other classic alternatives such as surgical excision with cryotherapy, which could induce the development of severe limbic deficiency in cases with extensive disease. PMID- 16372218 TI - [Optic nerve pilocytic astrocytoma with retinal involvement]. AB - INTRODUCTION: This is an atypical case of a pilocytic astrocytoma that involved the optic nerve (ON) and the retina. CLINICAL CASE: The patient was a 30-year-old male, who had attended ONCE since his early childhood because of the suspicion of an intraocular tumor. The ophthalmology exploration showed an ON and retinal coloboma in the right eye and microphthalmy, shutting of the pupil, retinal detachment and proof of an intraocular tumor in the left eye. The MR revealed an ON tumor that involved the retina. The histopathological study after enucleation was pilocytic astrocytoma. DISCUSSION: Long standing congenital pathology in a blind patient does not exclude the possibility of the patient suffering from other pathology as well. PMID- 16372219 TI - [Proptosis as the first sign of recurrent scirrhous breast carcinoma]. AB - OBJECTIVE: To inform clinicians of a metastatic orbital neoplasm that frequently masquerades as an orbital inflammatory syndrome. CASE REPORT: The case of a 61 year-old woman, diagnosed and treated seven years ago for a scirrhous breast carcinoma is presented. She complained of blurred vision and proptosis of the left eye and these symptoms were the first manifestation of tumor relapse. DISCUSSION: When orbital inflammation is detected in an adult female patient, a full medical history must be obtained and physical examination performed, as orbital metastases have been reported to be the initial manifestation of malignancy in 30 to 50% of cases. PMID- 16372220 TI - [Spontaneous resolution of an idiopathic epiretinal membrane in a young patient]. AB - CASE REPORT: Idiopathic epiretinal membranes are uncommon in young patients. The characteristics and evolution of these membranes are different from those in adults. We present the case of a young patient with an idiopathic epiretinal membrane, that several months later, showed spontaneous resolution with an associated posterior vitreous detachment. DISCUSSION: Idiopathic epiretinal membranes in young patients resolve spontaneously more frequently than in adults; however in many cases are associated with posterior vitreous detachment. We discuss the evolution of these membranes. PMID- 16372221 TI - [Fermin Munoz Urra and the experimental cataract. (1915)]. PMID- 16372222 TI - [Valentin Hauy (Saint Just-en-Chaussee, 1745-Paris, 1822)]. PMID- 16372223 TI - Diagnosis and management of tumors of the adrenal medulla. AB - The adrenal medulla consists of chromaffin cells, the site of catecholamine biosynthesis. Pheochromocytomas are chromaffin-cell tumors; 80-85 % arise from the adrenal medulla and 15-20 % arise from extra-adrenal chromaffin tissues (paragangliomas). Neuroblastomas are primitive tumors that derive from the same blastic precursor as in pheochromocytomas, and are distributed along the sympathetic nervous system. Pheochromocytomas account for 6.5 % of incidentally discovered adrenal tumors; they are found in 50 % of patients with multiple endocrine neoplasia 2A (MEN 2A) and 5-25 % of patients with von Hippel-Lindau (VHL) syndrome. Neuroblastomas are the most common solid extra-cranial tumors in children, and account for 7-10 % of all tumors. The diagnosis of pheochromocytoma should first be established biochemically by measuring plasma free metanephrines (the measurement of urinary fractionated metanephrines is the second choice). Measurements of homovanillic acid (HVA), norepinephrine and vanilmandelic acid (VMA) in urine are a necessity in patients with suspected neuroblastoma. Anatomical (radiological) imaging with computed tomography (CT) or magnetic resonance imaging (MRI) is necessary for both pheochromocytomas and neuroblastomas. Functional (nuclear medicine) methods are useful for both tumors. Scintigraphy with [123I]-metaiodobenzylguanidine is the specific functional imaging test of first choice; if this is not available, scintigraphy with [131I] MIBG is the second choice. Other newer specific modalities that have been used for evaluating pheochromocytomas include positron emission tomography (PET) with [18F]-F-fluorodopamine (F-DA) and [18F]-F-dihydroxyphenylalanine (DOPA). These should be used when MIBG scintigraphy is negative. Primary treatment for both types of tumor is surgical; chemotherapy is used for inoperable disease. After successful surgery, survival of patients with benign, sporadic pheochromocytomas is believed to be equal to that of the general population. Depending on the extent of disease and age, patients with neuroblastomas have cure rates of 15-90 %. PMID- 16372224 TI - Differential expression of the human somatostatin receptor subtypes sst1 to sst5 in various adrenal tumors and normal adrenal gland. AB - Somatostatin (SRIF) is a widely distributed peptide with growth-inhibiting effects in various tumors. So far, five distinct human SRIF receptor subtypes (sst1-sst5) have been identified. We investigated expression of the five ssts in various adrenal tumors and in normal adrenal gland. Tissue was obtained from ten pheochromocytomas (PHEOs), nine cortisol-secreting adenomas (CPAs), eleven aldosterone secreting adenomas (APAs) and eight non-functional adenomas (NFAs) after retroperitoneoscopic surgery, and used for RNA extraction. Adrenal tissue surrounding the tumor was available for analysis in twenty-seven cases. Receptor expression was studied by RT-PCR using sst-specific primers and subsequently confirmed by Southern blotting. Expression of all five receptor subtypes was observed in RNA obtained from normal adrenal gland. Furthermore, each receptor subtype was expressed in more than 50 % of all tumors analyzed. No sst5 expression was found in PHEOs, while sst1 was present in nearly all of these tumors. Only a few of the CPAs expressed subtypes sst1 and sst4. Expression of all five subtypes was distributed equally in APAs. No sst4 was found in any of the NFAs. Differential expression of ssts in various adrenal tumors may point to new aspects in the pathogenesis of these adenomas. Furthermore, the presence of specific ssts could expand the diagnostic and therapeutic strategies during management. New subtype specific analogues of SRIF may be used in the future depending on the type of adrenal tumor and receptor subtype expressed. PMID- 16372225 TI - Influence of oxygen concentration on redox cycling of alloxan and dialuric acid. AB - Alloxan, a chemical diabetogen, decays in the absence of reductants into alloxanic acid. In the presence of glutathione, it is reduced via the alloxan radical into dialuric acid, which autoxidizes back to alloxan. During this redox cycling process, reactive oxygen species are formed that destroy beta-cells in islets of Langerhans. Previous experiments were conducted with oxygen concentrations about ten times as high as within cells. The aim of our in vitro study was to evaluate the impact of different oxygen concentrations (0, 25, 250 micromol/l) at a given initial ratio of glutathione and alloxan on this redox cycling. Reduction of alloxan, oxidation of glutathione, and the formation of glutathiol (GSSG) were continuously recorded by HPLC for 90 minutes at 25 degrees C in air, calibration gas, or argon. In the absence of reductants, alloxan irreversibly decomposed into alloxanic acid regardless of oxygen presence. When the reaction system contained glutathione, decomposition was significantly retarded and therefore influenced by oxygen. In argon, decay could not be observed due to its reduction and the absence of oxygen. Increasing oxygen concentration enabled a redox cycling and therefore an ongoing decay. The highest decomposition along with the highest consumption of glutathione occurred at 250 micromol/l oxygen. The lower the oxygen, the more dialuric acid could be detected. After calculation, about 33 redox cycles per hour generates an amount of reactive oxygen species sufficient to damage pancreatic beta cells and induce insulin deficiency. PMID- 16372226 TI - No difference in lipolysis or glucose transport of subcutaneous fat cells between moderate-fat and low-fat hypocaloric diets in obese women. AB - The objective of the present study was to evaluate the effect of two different diets on lipolysis and lipogenesis in subcutaneous fat cells from obese women. In a ten-week nutritional intervention study, forty women were randomly assigned to a hypoenergetic-2,514 kJ (- 600 kcal/day) diet of either moderate-fat/moderate carbohydrate or low-fat/high-carbohydrate content. Body weight was equally reduced by approximately 7.5 % in both diet groups (p = 0.58). A subcutaneous adipose tissue biopsy was obtained for subsequent measurement of triglyceride breakdown (lipolysis) using drugs active at different steps of the lipolytic signaling cascade, and lipid synthesis (glucose transport) before and after intervention. No difference was found between the two diet groups at the maximum rate of either lipolysis or adrenoceptor sensitivity (p-values: 0.14 - 0.97). Inhibition of lipolysis by insulin was also similar in both diet groups before and after intervention. Finally, insulin-stimulated glucose transport did not show any changes that could be attributed to the type of diet. In conclusion, our data suggest that macronutrient diet composition has no major influence on glucose transport or mobilization of triglycerides in human subcutaneous fat cells of obese women. PMID- 16372227 TI - Relevance of TSH receptor stimulating and blocking autoantibody measurement for the prediction of relapse in Graves' disease. AB - Recently, we demonstrated that higher levels of autoantibodies to the human TSH receptor (TBII) predict relapse of hyperthyroidism in Graves' disease (GD). The aim of this study was to extend this outcome prediction by dividing TBII into stimulating (TSAb) and blocking (TBAb) TSH receptor autoantibodies. Altogether, ninety patients (81 female, 9 male) were retrospectively analyzed; sixty-four patients (71 %) did not go into remission or relapsed, whereas twenty-six patients (29 %) went into remission (median follow-up: 17.5 months). TSAb and TBAb measurement was performed in a CHO cell bioassay with cAMP readout at the time of their first visit in our outpatient clinic (single point measurement in median 6.5 months after initial diagnosis). In the remission group, eighteen of twenty-six patients (69 %) were TSAb-positive, whereas fifty-three of sixty-four patients (83 %) were TSAb-positive in the relapse group (p = ns). The mean stimulation indices (SI) were 4.1 in the remission group and 12.9 in the relapse group, respectively (p = 0.015). By using a threshold of 10 SI, the specificity for relapse was 96.0 %, as only one in twenty patients with an SI above 10 went into remission during follow-up (PPV 95 %). Most TSAb-positive patients also had high levels of TBII. Neither group showed any difference with respect to blocking type autoantibodies, which were mostly negative in both groups. In summary, high TSAb levels are similar but not superior to TBII for predicting relapse in GD patients. In contrast, TBAb measurement does not add any valuable information in this context. In the clinical routine, TSAb/TBAb measurement may not play an important role for diagnosis or outcome prediction of GD, since sensitive 2 (nd) generation TBII assays are easier to perform and offer similar information to the clinician. Bioassays should be reserved for special clinical questions such as Graves' disease in pregnancy. PMID- 16372228 TI - Binding, stimulating and blocking TSH receptor antibodies to the thyrotropin receptor as predictors of relapse of Graves' disease after withdrawal of antithyroid treatment. AB - TSH-receptor autoantibodies (TRAbs) are a valuable diagnostic tool for confirming a diagnosis of Graves' disease (GD). While there is evidence that high TRAb levels are associated with relapse of GD, whether a discrimination of TRAb into stimulating (TSAb) and blocking (TBAb) autoantibodies would benefit the clinician in terms of outcome prediction remains unclear. To address this issue, we have determined TRAb, TSAb and TBAb levels in serum samples of ninety-six euthyroid patients with GD taken four weeks after antithyroid drug withdrawal (ATDT). Forty seven patients (49 %) underwent relapse of GD within two years. Amongst those, forty-one (87 %) had been positive for TRAb and thirty-five (74 %) for TSAb after treatment. All patients except one were negative for TBAb. The correlation between TRAb and TSAb in those treated GD patients was relatively weak (r = 0.268, p < 0.001). Based on a cut-off limit of 1.5 IU/l, the positive and negative predictive values with respect to prediction of relapse were too low for any clinical relevance (TRAb: 49 % and 54 %; TSAb: 51 % and 55 %). However, when a cut-off level above 10 IU/l was used, the positive and negative predictive values increased to 83 % and 62 %. The additional measurement of TSAb or TBAb in those samples after therapy did not add additional information, even at higher decision thresholds. In conclusion, differentiation of TRAb into TSAb and TBAb is of no help in the prediction of relapse of GD in euthyroid patients at the end of ATDT, and only high TRAb levels are associated with relapse. PMID- 16372229 TI - Changes of thyroid function during long-term hGH therapy in GHD children. A possible relationship with catch-up growth? AB - BACKGROUND: Growth hormone (GH) treatment in patients with GH deficiency (GHD) can determine changes in the thyroid function. The clinical significance of these changes remains controversial, and all studies have so far covered rather a short period--usually no longer than one year. OBJECTIVE: To determine the effect of long-term recombinant hGH treatment in children with idiopathic GHD on the thyroid function. PATIENTS AND METHODS: Nineteen prepubertal children (12 boys and 7 girls, mean age 9.2 +/- 3.1 years) with idiopathic GHD were studied and followed for twenty-four months. None of the patients showed multiple pituitary hormone deficiencies. Nineteen healthy children matched for age and sex acted as controls. RESULTS: Patients with GHD showed a significant increase in TT (3) at twelve months and in FT (3) at six and twelve months after starting GH treatment, with a significant decrease at eighteen and twenty-four months. TT (4) level decreased significantly at twelve months and increased significantly at eighteen and twenty-four months. FT (4) also decreased, but only slightly, after twelve months of hGH treatment, and then increased significantly at twenty-four months. TSH levels did not vary significantly during the course of therapy. TT (3)/TT (4) and FT (3)/FT (4) ratios increased significantly after six and twelve months of therapy and significantly decreased later, approaching pre-therapy values. The SDS of Growth Velocity (SDS-GV) increased remarkably during the first year of therapy and then decreased significantly during the second year, although it remained significantly higher than the pre-therapy values. TT (3) and TT (3)/TT (4) ratio displayed a significant correlation with SDS-GV at twelve months of therapy. In a multiple regression analysis with age, bone age, parental height, GH dose, TT (3,) TT (3)/TT (4), and the SDS of IGF-I, only the TT (3)/TT (4) ratio at twelve months of therapy (p < 0.001) was identified as a significant predictor of SDS-GV. CONCLUSION: Our data confirm that changes in thyroid function are present in GHD children during long-term hGH therapy. These changes probably resulted from the effect of hGH on the peripheral metabolism of thyroid hormones and appear to be transitory, disappearing during the second year of hGH treatment. We speculate on the functional significance of these changes, and in particular, on their role in catch-up growth after hGH therapy. PMID- 16372230 TI - The intellectual capacity of patients with Laron syndrome (LS) differs with various molecular defects of the growth hormone receptor gene. Correlation with CNS abnormalities. AB - The correlation between the molecular defects of the GH receptor (R), psychosocial development and brain abnormalities were evaluated in 10 patients with Laron syndrome (LS), in whom all data were available. The findings revealed that the intelligence quotient (IQ) and abnormalities in the brain of the patients with LS differ with various molecular defects of the GH-receptor. The most severe mental deficits and brain pathology occurred in patients with 3, 5, 6 exon deletion. Patients with point mutations in exons 2, 4 and 7 presented various degrees of medium to mild CNS abnormalities that correlated with the IQ. Notably, the patient with the E180 splice mutation in exon 6 had a normal IQ, which fits the report on normal IQ in a large Ecuadorian cohort with the same mutation. This is the first report to support a correlation between IQ, brain abnormalities and localization of the molecular defects in the GH-R gene. As all patients with LS are IGF-I-deficient, it must be assumed that other as yet unknown factors related to the molecular defects in the GH-R are the major cause of the differences in intellect and brain abnormalities. PMID- 16372231 TI - Adipokine expression profile in adipocytes of different mouse models of obesity. AB - Adipose tissue produces and secretes multiple adipokines. Most studies on adipokine production/expression have been performed on whole adipose tissue. In addition, data concerning an overall of adipokine expression are scarce and can be heterogeneous depending on the obesity model studied. Our first aim was to compare the expression of adipokines involved in the interplay between obesity and insulin resistance in isolated adipocytes from different mouse models of obesity displaying different levels of weight gain and insulin sensitivity. The second aim was to determine perigonadal/subcutaneous ratio of each adipokine. Only resistin expression was decreased in obese mice without modifications in glucose and insulin blood levels. In addition to decreased levels of resistin, obesity models associated with hyperglycemia and hyperinsulinemia presented an increased expression of leptin and tumor necrosis factor-alpha (TNFalpha). Obese and diabetic mice were the only animals to exhibit high expression of plasminogen activator inhibitor type-1 and interleukin-6. All adipokines except TNFalpha were more heavily expressed in perigonadal than in subcutaneous adipocytes. Interestingly, fat-enriched diet and overweight on their own did not modify the distribution of adipokines between the two fat depots. However, severe obesity modified the distribution of proinflammatory adipokines. In conclusion, the level and number of adipokines with altered expression increased with obesity and hyperinsulinemia in mice. The physiopathological impact of depot-specific differences of adipokine expression in adipocytes remains to be clarified. PMID- 16372232 TI - Free fatty acids do not acutely increase asymmetrical dimethylarginine concentrations. AB - Concentrations of asymmetrical dimethylarginine (ADMA) and free fatty acids (FFAs) are elevated in insulin resistance which is associated with impaired vascular function. We hypothesized that FFAs could alter vascular tone by affecting ADMA concentrations. Plasma FFA levels were increased in seventeen healthy male volunteers by Intralipid/heparin infusion; hemodynamic and biochemical parameters were measured after 90 minutes. Plasma collected before and during Intralipid/heparin or equivalent synthetic FFAs was incubated with human umbilical vein endothelial cells (HUVECs) in vitro. Intralipid/heparin infusion resulted in an approximately seven-fold increase in plasma FFA levels to 1861 +/- 139 micromol/l, which was paralleled by increased systemic blood pressure and forearm blood flow. Intralipid/heparin did not affect ADMA (baseline mean 0.59 [95 % confidence interval [CI]: 0.54; 0.64] and 0.56 [CI: 0.51; 0.59] after 90 minutes), but slightly decreased SDMA (from 0.76, [CI: 0.70; 0.83] to 0.71 [CI: 0.64; 0.74], p < 0.05), and had no effect on ADMA/SDMA ratio. There was no correlation between ADMA and FFA concentrations or forearm blood flow. Incubation of HUVECs with FFA-rich plasma or synthetic FFAs induced an ADMA release after 24 hours, but not after 90 minutes. Acutely increased FFA levels caused hemodynamic effects but did not affect ADMA. Prolonged elevation of FFA levels might influence vascular function by increasing ADMA levels. PMID- 16372233 TI - Acute effects of high-protein and high-fat isoenergetic meals on total ghrelin plasma concentrations in lean and obese women. PMID- 16372234 TI - Reciprocal glycerolipid syntheses in skeletal muscle of obese rats. PMID- 16372235 TI - A correlational study on health training program participation and influences upon communities volunteers in Taipei City. AB - This is a cross-sectional study that purposely selected healthy community centers (HCC) in Taipei City to explore factors affecting volunteers' health training program participation (HTPP). The major objectives were to: (1) examine volunteer's HTPP; (2) explore relationships among volunteers' HTPP, self-efficacy (SE) in healthy community building, and community activities participation (CAP); and (3) identify key factors affecting volunteers' HTPP. A self-developed instrument with validity (content validity index > .91) and reliability (alpha = .63-.87) was used. A total of 250 participants were recruited. Study results revealed that a majority of the volunteers were middle-aged women who held at least a high school education, worked part-time, held Buddhist religious beliefs, were married and were middle class in terms of income. The average HTPP value was 19, with Xin Yi, Zhong Zhen and Bei Tou HCCs exhibiting higher HTPP values. Those not currently employed and housewives volunteers had lower HTPP values. 66.4% of volunteers participated at community activities after training and contributed 60 120 service hours a year. Volunteers' SE in healthy community building was low (M = 2.01, SD = 1.01), which, however, could explain 21% variance of HTPP. Factors affecting HTPP included HCC site, SE and CAP, which accounted for 57% of variance. Study results indicated that SE and CAP could serve as training evaluation indicators and a well-organized training program could increase volunteers' SE. Standard health training programs should incorporate three programs, including volunteerism perception, health promotion, and middle aged and elderly care to thus further improve volunteers' participation in community health activities. PMID- 16372236 TI - Clinical follow-up study on diabetes patients participating in a health management plan. AB - The "Health Management Plan"(HMP) for caring diabetic patient was begun by the National Health Insurance (NHI) in Taiwan in order to maximize the effectiveness of limited medical resources. This study describes the clinical experience of the nurse-directed diabetic HMP program and analyzes factors associated with the outcome. One hundred and thirty-six patients, all of whom had participated in the diabetic HMP program at least 5 times, were enrolled in the study. The effect of the HMP was evaluated by comparing hemoglobin A1C status before and after %) maintained their hemoglobin A1C status over the course of HMP participation. Diabetic patients with regular exercise habits showed a 2.8-fold increased chance of outcome improvement compared with those who did not exercise regularly. The chance of outcome improvement in patients with complications was found to be one third that of patients who had no complications (Odds ratio: 0.3, 95% CI: 0.1 1.0). This study found that specially trained nurses, following agreed upon protocols and algorithms and collaborating with medical team members, can effectively concentrate on providing comprehensive and effective diabetes care. PMID- 16372237 TI - Enema prior to labor: a controversial routine in Taiwan. AB - While taking an enema to induce labor is a controversial issue worldwide, in Taiwan it remains a routine procedure in many hospitals in preparation for birth. Episiotomy is also a prevalent procedure performed during the birthing process. Some physicians believe that enemas help reduce the risk of feces contamination of the episiotomy incision and, therefore, are justified as a routine procedure. This study compared the neonatal infection rates, times to appearance of fetal head, times to first post-labor bowel movement, and rates of episiotomy dehiscence of women receiving a pre-labor enema against those who did not. A total of 534 women classified with low-risk pregnancies were recruited from a medical center in central Taiwan and assigned randomly into one of two groups for a six-month period. The first group (264 subjects) received routine enema procedures prior to delivery in the first 6 months. The second group (270 subjects) did not receive enemas. Study results revealed no significant difference between enema and non-enema groups in terms of infection rates in mothers or infants or in terms of average time to fetal head appearance. While labor duration was the same for the two groups in the first and third stages of labor, the enema group experienced a relatively shorter second stage. No significant difference was observed in times to first post-labor bowel movement or episiotomy dehiscence rates. The results of this study indicate that the administration of enemas as a routine practice prior to labor is not substantiated by medical necessity. However, limitations of the research design suggest that a randomized clinical trial be adopted in the future to explore further the scientific validity of study results. PMID- 16372238 TI - Psychological abuse behavior exhibited by caregivers in the care of the elderly and correlated factors in long-term care facilities in Taiwan. AB - This study explored the state and the correlated factors of psychological abuse behavior among caregivers in long-term care facilities in Taiwan. A cross sectional design utilizing a descriptive-correlational approach was conducted. A sociodemographic sheet, Caregiver Psychological Elder Abuse Behavior Scale (CPEAB), and Caregiver's Burden Scale (CBS) were employed to collect data. A total of 114 caregivers completed the study, of whom 16.1% returned an CPEAB score greater than 40. The potential range of CPEAB scores ranged from 20 to 80, with the highest score indicating the greatest tendency toward psychological abuse behavior. Among the total of 20 CPEAB indicators, "Accuse him/her verbally", "Ignore his/her request", and "Insult him/her" received the highest item mean scores (M = 2.18, 2.12, 2.02; SD = 0.83, 0.84, 0.91). Significant relationships between the CPEAB and subject age, professional status, educational level, geriatric care training, and CBS score were found (p < .05 - .001). These results indicate that caregivers who are younger in age, less educated and lacking in geriatric training had a greater burden and displayed a tendency toward more abusive behavior. Another important finding was that nurses displayed more abusive behavior than their care attendant counterparts. Results of this study provide important information that may be used in medical and social interventions and policies aimed at improving quality of life for the institutionalized elderly. PMID- 16372239 TI - Physical activities and correlates of clinical nurses in Taipei municipal hospitals. AB - The purpose of this study was to investigate the physical activity profiles of clinical nurses in Taipei municipal hospitals. This study also explored the factors that influence physical activity, including personal background and social psychological factors. A total of 400 Taipei municipal hospital clinical nurses were selected using a randomized procedure to participate in a 3-day physical activity record assignment and fill in a structured questionnaire. Study results indicate that energy expenditure for clinical nurses on working days are higher than that on non-working days. Non-working days of clinical nurses are characterized by a lack of physical activity. Nurses who were required to take turns working in three shift rotation cycles, those with education at the junior college level or below, and those who were married tended to spend more energy on physical activity. Social psychological factors affecting the physical activities of clinical nurses include mainly exercise self-efficacy. Heavy work loading, poor health conditions, laziness, and lack of time tended to lower exercise self efficacy. On non-working days, perceived barriers to exercise are the main issue area affecting moderate-to-vigorous physical activity. Nurses with greater perceived barriers to exercise spend less energy on physical activity. The main influencing factors for perceived barriers to exercise include laziness and lack of time. PMID- 16372240 TI - Constructing and testing a model of trustworthiness, trust behavior and organizational identification. AB - This study, in which staff nurses in Taiwan participated, was aimed to construct a model for relationships among trustworthiness, trust behavior and organizational identification. One hundred and ninety five anonymously completed questionnaires were collected. Superiors' trustworthiness, staff nurses' trust behavior, and organizational identification were identified as latent variables to which the following indicators were to be applied: integrity, benevolence, ability, communication, consistency, compliance, sharing, teamwork, subordination, membership, loyalty and similarity. A logical trust and organizational identification model was constructed and tested with LISREL (Linear Structural Relations) analysis. Benevolence was eliminated due to excessive residual value. The adjusted model passed pertinent tests. It was also revealed that trust behavior acts as a mediator between trustworthiness and organizational identification. The results of this study may be taken as a reference for the training of supervisory level personnel, as well as for strengthening the organizational identification of staff nurses, and as a beginning phase in the construction and testing of a relationship model for the relationship between trust and organizational identification. PMID- 16372241 TI - The health status and health promotion behavior of low-income elderly in the Taipei area. AB - The purpose of this study was to understand health promotion behaviors and their correlated factors among low-income elderly in the Taipei area. This study used a cross-sectional design with a descriptive correlation approach. A total of 89 low income elderly from Peitou District of Taipei participated in the study. Structured questionnaires, the primary sources of measurement, were read to study subjects by a trained research assistant. The mean age of low-income elderly in the study was 76.85 year-old. A plurality were single (41.6%) and 31.5% lived alone. Subject IADLs (instrumental activities of daily living) were not as good as their ADLs (activities of daily living). Their psychosocial health status was worse than their physical health status. Among health promotion behaviors, regular exercise appears to be the most prevalent. The most significant factor among subjects for failing to follow health-promoting behaviors was their IADLs. Homebound elderly who lived alone and had poor IADL were less likely to have health promoting behaviors. A proposal to encourage health promotion behaviors for low-income elderly derived from study results is the initiation of formal personal assistance programs to remedy the inability of many elderly, especially those living alone, to leave the home due to ADL-related or other limitations. PMID- 16372242 TI - Clarification of the antecedents of hospital nurse organizational citizenship behavior -- an example from a Taiwan regional hospital. AB - Organizational citizenship behaviors (OCB) are deemed indispensable due to their importance in promoting positive relationships among employees and involving employees in the organization's activities. OCB is believed to facilitate attainment of a hospital's goals and enhance a hospital's performance. The aim of this study is to extend the work of previous studies by providing an integrated model of OCB and demonstrate its applicability in a non-Western culture. Questionnaires were completed by 314 non-supervisory hospital nurses, each of whom was given a supervisor rating for their OCB. Exploratory factor analysis was used to assess the convergent and discriminant validity of the variables in the model. Multiple regression analysis was used because it provided estimates of net effects and explanatory power. The results showed that job satisfaction, supervisor support, job involvement, and procedural justices had significant effects on the nurses' citizenship behaviors. Since the study used a non-Western site in one of Taiwan's health care institutions, its overall findings should contribute to further understanding of the cross-cultural aspects of OCB. PMID- 16372243 TI - Microplicae-like structures of the fallopian tube in postmenopausal women as shown by electron microscopy. AB - Female reproductive aging is associated with several morphological changes of the genital tract with a subsequent decline in fertility; however, ultrastructural changes occurring after menopause have still not been well illustrated. Our aim was to characterize the three-dimensional microanatomy of the luminal surface of the human fallopian tube in perimenopause and postmenopause. Twenty bioptic samples of fallopian tubes were obtained after surgery under the informed consent of the patients. Samples were processed for transmission electron microscopy (TEM) and field-emission scanning electron microscopy (FE-SEM). As age increases the surface epithelium of the fallopian tube appeared somewhat flattened. Correlated TEM/FE-SEM observations showed gradual shortening of microvilli and deciliation. The most interesting finding was the gradual formation of microplicae-like structures on the surface epithelium, particularly from late perimenopause to postmenopause. Microplicae-like structures, associated with other regressive changes, represent an important adaptation of the epithelium of the fallopian tube; these are likely induced by the physiological process of aging, thus better withstanding hormonal changes associated with the advent of the menopause. PMID- 16372244 TI - HER-2 status determination in breast carcinomas. A practical approach. AB - Accurate evaluation of HER-2 status is crucial in the selection of breast carcinoma patients for trastuzumab (Herceptin) treatment. Various laboratory methods have been used for this purpose. The aim of the present work was to analyse the results obtained in the routine practice by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in determination of HER-2 status. Five hundred and three cases of breast invasive ductal carcinoma were selected to analyse the HER-2 overexpression by immunohistochemistry (HercepTest, Dako). HercepTest 2+ equivocal cases (60) were studied by FISH (PathVysion, Vysis) to determine HER-2 gene amplification. HER-2 overexpression determined by Herceptest was shown in 97/503 cases (19%). FISH performed on equivocal cases demonstrated HER-2 amplification in 11/60 tumours (18%). IHC and FISH together showed HER-2 overexpression/gene amplification in 21% of breast invasive carcinomas. Immunohistochemical determination of HER-2 status represents an easy and standardized method that (in contrast to FISH) can be performed in all pathology laboratories without need of any special microscope and enabling to check the morphologic features of the cells analysed. However, in order to assure the reliability of the results, standardization of fixation protocols, automation of the immunohistochemical procedure, and training of pathologists in the interpretation of the results (scoring criteria) should be a priority. Equivocal HercepTest cases must be analysed by FISH preferably in a reference laboratory. PMID- 16372245 TI - Basement membrane heterogeneity during chick development as shown by tomato (Lycopersicon esculentum) lectin binding. AB - Basement membranes (BMs) constitute a distinct compartment of the extracellular matrix (ECM). All BMs show a similar structural appearance but differ in molecular composition. These variations have critical functional implications. The aim of this study is to establish the pattern of the tomato lectin (Lycopersicon esculentum agglutinin--LEA) binding sites in the BMs of the developing chick embryo (stages 4-21, Hamburger and Hamilton, 1951) in order to achieve a better understanding of the molecular heterogeneity of BMs. The study was performed with transmission electron microscopy (TEM) histochemistry, and confocal laser microscopy. TEM showed that LEA bound to the lamina densa and to the lamina fibroreticularis of the BMs. Through the period studied, most of the LEA binding appeared in the ectodermal BM and its derivatives. In the limb bud, LEA binding to the ectoderm BM was more intense in the ventral half than in the dorsal half. Furthermore, LEA allowed the early (HH16) detection of the transverse fibrillar tracts. In the lens and in the inner ear primordium, the BMs were LEA positive through the placode and cup stages. The binding was progressively reduced through the vesicle stage. The BMs of the olfactory primordium, and of the Rathke's pouch were positive. In contrast, the BMs of the developing central nervous system were negative. The BMs of both the paraxial and the lateral plates of the mesoderm were negative, whereas the notochord and the BM of the Wolffian duct were positive. The endodermal BM and its derivatives were negative. The ECM located between the fusing endocardial tubes, and the BM of the fusion zone of the paired aortae, were positive. This suggested an active role of the LEA-positive glycoproteins in the fusion of endothelia. Our results show the heterogeneity of the chick embryo BMs during development. In addition, LEA constitutes an excellent marker for the primordial germ cells. PMID- 16372246 TI - Immunoexpression of the CD30 ligand/CD30 and IL-6/IL-6R signals in thyroid autoimmune diseases. AB - To elucidate the role of Th2 cytokines in autoimmune thyroid diseases, we have studied by immunohistochemistry the expression of two Th2 ligand/receptor systems (CD30-L/CD30 and IL-6/IL-6R) in goitrous Graves' disease (GD) and Hashimoto's thyroiditis (HT). A total number of 50 nodular goiters (NG), including 10 GD showing a lymphoid aggregate grade I, 30 HT 8 of which had a lymphoid aggregate of grade I, 12 of grade II and 10 grade III, and 10 colloid goiters have been evaluated. In addition, 5 normal thyroids were included in the study as controls. Reactivity of ligand and cognate receptor of both CD30-L/CD30 and IL-6/IL-6R pathways was observed in a greater proportion of GD, compared to HT (P<0.005). In HT, the expression of CD30-L/CD30 system was detected more frequently than IL 6/IL-6R (P<0.05) and showed an inverse correlation with the grade of lymphoid aggregate, whereas IL-6/IL-6R correlated positively with lymphocyte infiltration (P<0.05). Based on our results concerning a dominance of Th2 cytokines in GD, we postulate that CD30-L/CD30 and IL-6/IL-6R systems could play a major role in the pathogenesis of GD. However, the expression of CD30L/CD30 and IL-6/IL-6R in HT suggests that Th2 mechanisms are involved also in tissue damage of HT. The two systems could contribute to drive the autoimmune response skewing toward a Th2 phenotype and this appears to be correlated with the lymphoid aggregate grade. PMID- 16372247 TI - Histopathological changes in the brain of mouse fetuses by etoposide administration. AB - Etoposide (VP-16), a topoisomerase II inhibitor, is an anti-tumor agent which is also known to show embryotoxicity, and teratogenicity when administered to pregnant rodents. We examined VP-16-induced histopathological changes in the brain of mouse fetuses. Pregnant mice were intraperitoneally injected with VP-16 (4 mg/kg) on day 12 of gestation (GD 12), and fetuses were collected from 1 to 48 hours after treatment (HAT). Mitotic neuroepithelial cells in the telencephalic wall prominently decreased at 2 HAT, and were hardly observed at 4 HAT. The number of pyknotic neuroepithelial cells in the fetal brain began to increase at 4 HAT, and became prominent from 8 to 24 HAT. These pyknotic cells were also positively stained by TUNEL method, which can detect fragmented DNA, and showed ultrastructural characteristics of apoptosis. Additionally, these cells were also positive for cleaved caspase-3, an essential executioner of apoptosis. This indicated that excessive neuroepithelial cell apoptosis was induced in the brain of mouse fetuses following VP-16 treatment on GD 12. PMID- 16372248 TI - Application of in vivo cryotechnique to the examination of cells and tissues in living animal organs. AB - When all biological materials in cells and tissues of living animal organs are quickly and promptly frozen, immunolocalization of their components and structural features in situ is necessary to understand their in vivo functioning states. However, these direct morphological analyses were difficult to achieve by conventional chemical fixation methods during the last century. A new cryofixation method, named the "in vivo cryotechnique", in which the normal blood circulation in living animals is always retained at the moment of freezing, has become a powerful tool to visualize the real native morphology of cells and tissues with functional meaning. The "in vivo cryotechnique" can usually be combined with a wide range of subsequent preparation techniques, and can thereby enable us to perform various direct analyses on biological samples, reflecting the physiological functions of living animal organs. PMID- 16372249 TI - Gut-trophic feed additives and their effects upon the gut structure and intestinal metabolism. State of the art in the pig, and perspectives towards humans. AB - The correct functional development of the gastrointestinal tract is of special importance during the neonatal and weaning phases of reared piglets. Nutrition is obviously a critical determinant in the growth of the gut in the young swine. The mucosal epithelium of the small intestine is reputed anatomically and functionally immature in neonatal pigs, a feature that appears to be exacerbated at weaning, when a colonization of the gut occurs by "new" microorganisms entering the alimentary canal with the solid feed. This frequently exposes piglets to diarrhoeic syndromes and other intestinal disturbances. Functional feed additives, also called nutraceuticals, appear as promising alternative substances to the use of chemotherapeutics as growth promoters in the rearing farm, above all considering the near banning of them by the European Parliament in the view of reducing antibiotic resistance phenomena in human therapies. Several feed additives are available that may play a role in the pig nutritional plan because of their trophic and cyto-protective effects on the gastrointestinal apparatus. Paying special attention to the quantitative consequences (histometry) upon the gut of the examined dietary supplements, this review, even if not fully exhaustive, will focus on the function (and possibly the mechanism/s of action) of certain gut-trophic nutrient substrates. This in turn will sustain the potential use of these substances in human therapy, especially the one directed at resolving intestinal diseases, both in adult and infant ages. In nutritional studies as well as in other biomedical research fields, the swine is an excellent animal model. PMID- 16372250 TI - Suppression of growth of tumour cell lines in vitro and tumours in vivo by mistletoe lectins. AB - A variety of studies have shown that incubation of different tumour cell lines with mistletoe lectins (MLs) in vitro has a marked cytotoxic effect. In the concentration range of low cytotoxicity cell death induced by ML-I is quantitatively due to apoptotic processes. The first events observed being membrane perforation and protusions. Simultaneous treatment of certain tumour cells with MLs rendered them more sensitive to induction of apoptosis by TNFalpha. The immunomodulatory activity of ML-I was investigated by measuring cytokine release and the results confirmed that cytokine induction by the lectin is regulated at the transcriptional level. ML-I has been shown to potentiate the effect of chemotherapeutic drugs. In addition to an in vitro effect a number of workers have demonstrated that MLs suppress tumour growth in vivo. Mistletoe lectins have been administered to animals locally to the tumour, systemic, subcutaneously or by the oral route via the diet. In many cases apoptosis was observed in the tumour and instances where complete tumour ablation has occurred have been reported. It has been hypothesized that the anticancer efficacy of tumour necrosis factor-alpha (TNFalpha) is potentiated by MLs isolated from both European and Korean mistletoe. There is accumulating evidence that both types of MLs are able to induce an anti-angiogenic response in the host suggesting that the anti-metastatic effect observed on a series of tumour cell lines in mice is in part due to an inhibition of tumour-induced angiogenesis and in part due to an induction of apoptosis. PMID- 16372251 TI - Exploring cyclosporine A-side effects and the protective role-played by antioxidants: the morphological and immunohistochemical studies. AB - Cyclosporine A (CsA) is the immunosuppressor most frequently used in transplant surgery and in the treatment of autoimmune diseases, because of its specific inhibiting effect on the signal transduction pathways of cell T receptor. It has been shown that CsA is able to generate reactive oxygen species and lipid peroxidation, which are directly involved in the CsA nephrotoxicity, hepatotoxicity and cardiotoxicity. So, the use of antioxidants seems to be a useful tool in attempting to reduce CsA adverse effects. The aim of this review is to summarise the general aspect of CsA, the classification of antioxidants, their mechanism of action and their administration for improving CsA side effects. The protective role of different antioxidants has been evaluated on CsA induced nephrotoxicity. It has been shown that the antioxidants, improved the morphological renal cytoarchitecture, increased the antioxidant enzyme content, decreased lipid peroxidation and reactive species oxygen (ROS). The protective role of antioxidants was also found in CsA hepatotoxicity and was related to the increase in antioxidant capacity of hepatic tissue, which was responsible for ameliorating hepatic morphology. Recently, it has been demonstrated that CsA induces side effects on the heart but the data to this purpose are very few and also the number of results on the protective role played by antioxidants it is very limited. In conclusion, not only do these observations provide insight into the intricate mechanism of CsA adverse effects, but they also present novel opportunities for the design and development of more effective therapeutic strategies against negative effects. PMID- 16372252 TI - Mechanisms underlying estrogen-induced sexual differentiation in the hypothalamus. AB - Estrogen plays critical roles in the sexual differentiation of the developing brain and gender-specific regulation of reproductive neuroendocrinology. Of the different regions of the brain, it is well known that hypothalamic areas contain key sexually differentiated neuronal circuits. Estrogen receptor (ER) proteins localized in the nucleus affect the expression of target genes when bound to their ligand estrogen. However, recent studies suggest that this may not be the only mechanism of estrogen action. Instead, estrogen can influence various cellular events through regulating different signaling pathways. Cross-talk between direct effects of estrogen on gene transcription and its effects on signaling pathways should be examined in future to elucidate mechanisms underlying sexual differentiation in the hypothalamus. PMID- 16372253 TI - Advances in molecular classification of renal neoplasms. AB - Kidney neoplasms are classified by light microscopy using the World Health Organization (WHO) system. The WHO system defines histopathologic tumor subtypes with distinct clinical behavior and underlying genetic mutations. In adults, the common malignant subtypes are variants of renal cell carcinoma (RCC). Histopathologic classification is critical for clinical management of RCC, but is becoming more complex with recognition of novel tumor subtypes, development of procedures yielding small diagnostic biopsies, and emergence of molecular therapies directed at tumor gene activity. Therefore, classification systems based on gene expression are likely to become essential for diagnosis, prognosis and treatment of kidney tumors. Recent DNA microarray studies have shown that clinically relevant renal tumor subtypes are characterized by distinct gene expression profiles, which are useful for discovery of novel diagnostic and prognostic biomarkers. In this review, we summarize the WHO classification system for renal tumors, general applications of microarray technology in cancer research, and specific microarray studies that have advanced knowledge of renal tumor diagnosis, prognosis, therapy and pathobiology. PMID- 16372254 TI - Radiological findings do not support lateral residual tumour as a major cause of local recurrence of rectal cancer. AB - BACKGROUND: The aim of this study was to determine the sites of local recurrence following radical (R0) total mesorectal excision (TME) for rectal cancer in an effort to elucidate the reasons for recurrence. METHODS: Thirty-seven patients with recurrence following curative resection for rectal cancer were identified from a population of 880 patients operated on by surgeons trained in the TME procedure. Two radiologists independently examined 33 available computed tomograms and magnetic resonance images taken when the recurrence was detected. RESULTS: Twenty-nine of the 33 recurrences were found in the lower two-thirds of the pelvis. Two recurrent tumours appeared to originate from lateral pelvic lymph nodes. Evidence of residual mesorectal fat was identified in 15 patients. Fourteen of the recurrent tumours originated from primary tumours in the upper rectum; all of these tumours recurred at the anastomosis and 12 of the 14 patients had evidence of residual mesorectal fat. CONCLUSION: Lateral pelvic lymph node metastases are not a major cause of local recurrence after TME. Partial mesorectal excision may be associated with an increased risk of local recurrence from tumours in the upper rectum. PMID- 16372255 TI - Letter: Randomized clinical trial of the effects of oral preoperative carbohydrates on postoperative nausea and vomiting after laparoscopic cholecystectomy (Br J Surg 2005; 92: 415-421). PMID- 16372256 TI - Affective judgment and beneficial decision making: ventromedial prefrontal activity correlates with performance in the Iowa Gambling Task. AB - Damasio proposes in his somatic marker theory that not only cognitive but also affective components are critical for decision making. Since affective judgment requires an interplay between affective and cognitive components, it might be considered a key process in decision making that has been linked to neural activity in ventromedial prefrontal cortex (VMPFC). Using functional magnetic resonance imaging (fMRI), we examined the relationship between VMPFC, emotionally (unexpected)- and cognitively (expected)-accentuated affective judgment, and beneficial decision making (Iowa Gambling Task; IGT) in healthy subjects. Neuronal activity in the VMPFC during unexpected affective judgment significantly correlated with both global and final performance in the IGT task. These findings suggest that the degree to which subjects recruit the VMPFC during affective judgment is related to beneficial performance in decision making in gambling. PMID- 16372257 TI - Detection of phosphorylation patterns in rat cortical neurons by combining phosphatase treatment and DIGE technology. AB - Although protein phosphorylation is probably the most studied post-translational modification occurring in cells, the number of proteins, which are the target of this modification, is still largely unknown. Increasing the coverage of the phosphoproteome as well as the detection of variation at the phosphorylation level would be very helpful for understanding the mechanisms of cell life and the modifications of the cell state leading to pathological conditions such as neurodegeneration. In order to further investigate variations occurring at the phosphorylation level, we have initiated the creation of a reference map of phosphorylated proteins in rat cortical neurons, employing a combination of phosphatase treatment and 2-DE/differential in gel electrophoresis technology. About 131 spots were recognized as phosphorylated proteins as they showed different migration behaviour after phosphatase treatment. The analysis of 42 selected spots was carried out by LC/MS/MS technology resulting in the identification of two new phosphoproteins. PMID- 16372258 TI - Chemical derivatization of phosphoserine and phosphothreonine containing peptides to increase sensitivity for MALDI-based analysis and for selectivity of MS/MS analysis. AB - Protein phosphorylation is one of the most important and common ways of regulating protein function in cells. However, phosphopeptides are difficult to analyse, ionising poorly under standard MALDI conditions. Several methods have been developed to deal with the low sensitivity and specificity of phosphopeptide analysis. Here, we show an approach using a simple one-step beta elimination/Michael addition reaction for the derivatization of phosphoserine and phosphothreonine. The substitution of the negatively charged phosphate group by a positively charged S-ethylpyridyl group greatly improves the ionisation of the modified peptides, especially in MALDI MS, increasing the sensitivity of the analysis. The modification allows the formation of a unique fragment ion at m/z 106 under mild collisional activation conditions, which can be used for parent (precursor) ion scanning in order to improve both the sensitivity and the selectivity of the analysis. The optimisation of the approach is described for a standard model peptide and protein and then applied to phosphorylation analysis in two biologically derived proteins purified from different experimental systems. PMID- 16372259 TI - DIGE compatible labelling of surface proteins on vital cells in vitro and in vivo. AB - Efficient methods for profiling of the cell surface proteome are desirable to get a deeper insight in basic biological processes, to localise proteins and to uncover proteins differentially expressed in diseases. Here we present a strategy to target cell surface exposed proteins via fluorescence labelling using CyDye DIGE fluors. This method has been applied to human cell lines in vitro as well as to a complex biological system in vivo. It allows detection of fluorophore-tagged cell surface proteins and visualisation of the accessible proteome within a single 2-D gel, simplifying subsequent UV MALDI-MS analysis. PMID- 16372260 TI - New insights into the pathological mechanisms of cerebrotendinous xanthomatosis in the Taiwanese using genomic and proteomic tools. AB - Cerebrotendinous xanthomatosis (CTX) is an autosomal recessive lipid-storage disorder caused by a deficiency of the mitochondrial sterol 27-hydroxylase. Genetic analysis utilizing SSCP and direct DNA sequencing identified a new mutation. One base-pair of cytosine was deleted at codon 326 on exon 2 of CYP27 in all CTX patients while their father was heterozygotic. This novel point deletion predicts a frameshift in mRNA (Pro(102) -->Leu) and results in the appearance of a premature termination codon (TGA) to substitute for Val(106) (GTG). To characterize the pathological mechanism of CTX patients, the protein profiles of serum and leukocytes extracted from these subjects were presented by means of proteomic technologies including 2-DE and MALDI-TOF analysis. According to the results, the amount of vinculin, ABP-280, talin and vimentin in leukocytes of CTX patients had changed significantly, reflecting the changes in membrane dynamics concerning cholestanol accumulation. The expression of target proteins in CTX patients and control was further verified by western blotting which indicated the same tendency as 2-DE data. This is the first paper to integrate both genomic and proteomic concepts for analyzing the possible mechanism of CTX and provides more information for related study in the future. PMID- 16372261 TI - Proteome analysis identifies novel protein candidates involved in regeneration of the cerebellum of teleost fish. AB - In contrast to mammals, adult teleost fish exhibit an enormous potential to regenerate neuronal tissue after injuries to the CNS. By combining a well-defined cerebellar lesion paradigm with differential proteome analysis at a post-lesion survival time of 3 days, we screened for protein candidates involved in repair of the fish brain. Out of nearly 900 protein spots detected on 2-D gels, spot intensity was significantly increased at least twofold in 30 spots and decreased to at least half the intensity of control tissue in 23 spots. The proteins associated with 24 of the spots were identified by PMF and MS/MS fragmentation. The cellular localization and the spatiotemporal patterns of two of these proteins, beta-actin and beta-tubulin, were further characterized through immunohistochemistry. Comparison of the observed changes in protein abundance with the previously characterized events underlying regeneration of the cerebellum suggests that the proteins identified are especially involved in cellular proliferation and survival, as well as axonal sprouting. PMID- 16372262 TI - Redox regulation of cyclophilin A by glutathionylation. AB - Using redox proteomics techniques to characterize the thiol status of proteins in human T lymphocytes, we identified cyclophilin A (CypA) as a specifically oxidized protein early after mitogen activation. CypA is an abundantly expressed cytosolic protein, target of the immunosuppressive drug cyclosporin A (CsA), for which a variety of functions has been described. In this study, we could identify CypA as a protein undergoing glutathionylation in vivo. Using MALDI-MS we identified Cys52 and Cys62 as targets of glutathionylation in T lymphocytes, and, using bioinformatic tools, we defined the reasons for the susceptibility of these residues to the modification. In addition, we found by circular dichroism spectroscopy that glutathionylation has an important impact on the secondary structure of CypA. Finally, we suggest that glutathionylation of CypA may have biological implications and that CypA may play a key role in redox regulation of immunity. PMID- 16372263 TI - Effects of oral vitamin C supplementation in hemodialysis patients: a proteomic assessment. AB - Evidence indicates that oxidative stress is present in dialysis patients, and is associated with vitamin C deficiency. Limited data are available regarding the effects of vitamin C supplementation on oxidative stress and inflammation markers in these patients. Moreover, there are no data available on plasma polypeptide fingerprints by proteome analysis before and after vitamin C supplementation. Therefore, we analyzed plasma samples from a prospective, randomized, open labeled trial to assess the effects of oral vitamin C supplementation (250 mg three times per week), to define the plasma polypeptide pattern in hemodialysis patients. Our results reveal that more than 30 polypeptides show significant changes in the dialysis patients in comparison to controls with normal renal function, and that several polypeptides are affected/normalized by oral vitamin C supplementation. These results underline the remarkable potential for proteomics to recognize specific peptide profiles in different pathological situations, which might not be detected by classical methods. PMID- 16372264 TI - Identification of vaccine candidate antigens of an ESBL producing Klebsiella pneumoniae clinical strain by immunoproteome analysis. AB - Klebsiella pneumoniae is an opportunistic pathogen which causes pneumoniae, urinary tract infections and septicemia in immunocompromised patients. Hospital outbreaks of multidrug-resistant K. pneumoniae, especially those in neonatal wards, are often caused by strains producing the extended-spectrum-beta lactamases (ESBLs). An immunoproteome based approach was developed to identify candidate antigens of K. pneumoniae for vaccine development. Sera from patients with acute K. pneumoniae infections (n = 55) and a control group of sera from healthy individuals (n = 15) were analyzed for reactivity by Western blot against ESBL K. pneumoniae outer membrane proteins separated by 2-DE. Twenty highly immunogenic protein spots were identified by immunoproteomic analysis. The immunogenic proteins that are most frequently recognized by positive K. pneumoniae sera were OmpA, OmpK36, FepA, OmpK17, OmpW, Colicin I receptor protein and three novel proteins. Two of the vaccine candidate genes, OmpA (Struve et al. Microbiology 2003, 149, 167-176) and FepA (Lai, Y. C. et al.. Infect Immun 2001, 69, 7140-7145), have recently been shown to be essential in colonization and infection in an in vivo mouse model. Hence, these two immunogenic proteins could serve as potential vaccine candidates. PMID- 16372265 TI - Analysis of outer membrane proteome of Escherichia coli related to resistance to ampicillin and tetracycline. AB - The elucidation of the molecular details of antibiotic resistance will lead to improvements in extending the efficacy of current antimicrobials. In the current study, proteomic methodologies were applied to characterize functional outer membrane proteins (Omps) of E. coli K-12 responded to tetracycline and ampicillin resistance for understanding of universal pathways that form barriers for antimicrobial agents. For this purpose, E. coli K-12 expressional outer membrane proteome was characterized and identified with the use of 2-DE and MALDI-TOF/MS methods. Then, differential Omps due to tetracycline or ampcilin resistance were determined by comparison between tetracycline minimum inhibitory concentration (MIC)10, ampicillin MIC10, control0 and control10, showing 9 proteins with 11 spots for tetracycline and 8 protein with 9 spots for ampicillin, showing a difference in only 1 protein (decreased LamB in tetracyclin) between the two antibiotics. Among the proteins, 3 were known as antibiotic-resistant proteins, including TolC, OmpC and YhiU, while FimD precursor, LamB, Tsx, YfiO, OmpW, NlpB were first reported here to be antibiotic-resistance-related proteins. Our findings will be helpful for further understanding of antibiotic-resistant mechanism(s). This study also shows that the combination of Omp purification methods certainly contributes the sensitivity of Omp detection. PMID- 16372266 TI - Expressed proteome analysis of human hepatocellular carcinoma in nude mice (LCI D20) with high metastasis potential. AB - We report for the first time an expressed proteome for human hepatocellular carcinoma (HCC) in nude mice model. Most cases of human liver cancer are HCC with highly metastatic ability. Therefore, the early prediction or diagnosis and effective treatment are the key points of research. We have previously successfully established a human HCC nude mice model (LCI-D20) with high metastasis potential. To understand better the tumor biology of HCC it is worth to explore the relativity of all expressed protein profiles in the LCI-D20 HCC nude mice model. With advanced proteomics technologies, we have carried out a proteomic analysis with following stages: protein sample preparation of cancer tissue, including total cellular extraction and sequential fractionation, 2-DE and 2-D LC separation, ESI/MALDI-MS/MS identification, as well as data-dependent bioinformatics. The identified proteins were classified bioinformatically respective to their function, biological process and intracellular localization. Some important proteins found in HCC, e.g. metabolism enzymes, proteins regulating cell motility, signaling proteins, and heat shock proteins, are discussed in terms of their metastasis. PMID- 16372267 TI - Proteomic analysis of the cerebrospinal fluid of patients with lumbar disk herniation. AB - To better understand the pathophysiologic mechanisms underlying spinal nerve root injury induced by lumbar disk herniation (LDH), comparative proteomic analysis of cerebrospinal fluid (CSF) between patients with LDH (the experiment group) and the otherwise healthy patients who had had implants removed from healed fractures in the lower limbs (the control group) was carried out using 2-DE followed by LC IT-MS and database searching. Image analysis of silver-stained 2-DE gels revealed that 15 protein spots showed significant differential expression between the two groups of CSF samples (p < 0.05). After searching the database we found that in CSF of LDH patients, the expression of cystatin C, apolipoprotein A-IV, vitamin D binding protein, neurofilament triplet L protein, IgG, tetranectin, and hemoglobin were elevated. However, ProSAAS, prostagladin D2 synthase, creatine kinase B, superoxide dismutase 1 and peroxiredoxin 2 were decreased. The subsequent ELISA measured the concentration of tetranectin, vitamin D-binding protein and cystatin C and confirmed the results of proteomic analysis. These identified proteins are involved in the pathophysiological process of spinal nerve root injury caused by herniated lumbar disk. The functional implications of the alterations in the levels of these proteins are discussed in this paper. PMID- 16372268 TI - Proteome analysis of early post-mortem changes in two bovine muscle types: M. longissimus dorsi and M. semitendinosis. AB - To study early post-mortem changes in muscle tissues from bull calves, cytosole proteins from two muscles: M. longissimus dorsi (LD) and M. semitendinosis (ST) at 0 and 24 h after slaughter were analysed by 2-DE. Principal component analysis (PCA) and rotation testing were used to analyse the protein patterns in the two muscles in order to select protein spots that were significantly different at the two time-points. Selected proteins were identified by MALDI-TOF/TOF. Five proteins, namely cofilin, lactoylglutathione lyase, substrate protein of mitochondrial ATP-dependent proteinase SP-22, HSP 27 and HSP20, were changed in both LD and ST muscles during post-mortem storage. Fifteen additional protein changes were observed in either LD or ST muscles, and some of these changes have not previously been observed to change during post-mortem storage of bovine muscles. Further studies will reveal the relevance of these biomarkers for meat quality. PMID- 16372269 TI - Proteome analysis of up-regulated proteins in the rat spinal cord induced by transection injury. AB - The inability of the CNS to regenerate in adult mammals propels us to reveal associated proteins involved in the injured CNS. In this paper, either thoracic laminectomy (as sham control) or thoracic spinal cord transection was performed on male adult rats. Five days after surgery, the whole spinal cord tissue was dissected and fractionated into water-soluble (dissolved in Tris buffer) and water-insoluble (dissolved in a solution containing chaotropes and surfactants) portions for 2-DE. Protein identification was performed by MS and further confirmed by Western blot. As a result, over 30 protein spots in the injured spinal cord were shown to be up-regulated no less than 1.5-fold. These identified proteins possibly play various roles during the injury and repair process and may be functionally categorized as several different groups, such as stress responsive and metabolic changes, lipid and protein degeneration, neural survival and regeneration. In particular, over-expression of 11-zinc finger protein and glypican may be responsible for the inhibition of axonal growth and regeneration. Moreover, three unknown proteins with novel sequences were found to be up regulated by spinal cord injury. Further characterization of these molecules may help us come closer to understanding the mechanisms that underlie the inability of the adult CNS to regenerate. PMID- 16372270 TI - Massspectrometrical analysis of recombinant human growth hormone Norditropin reveals amino acid exchange at M14_V14 rhGH. AB - Recombinant human growth hormone (rhGH) is used for the treatment of several disorders. Structural integrity of rhGH is of critical importance for its clinical use and modifications thereof may act as markers in situations such as rhGH doping, as illegal rhGH-abuse in sports is of increasing interest. In the current study we investigated homogeneity of Norditropin, a recombinant human growth hormone frequently used in medicine, expressed in E. coli, strain MC1061. The most recent proteomics technologies including 2-DE, MALDI-MS followed by MALDI-MS/MS and LC-MS followed by LC-MS/MS were used for the characterisation of rhGH. MALDI-TOF-TOF and electrospray LC-MS analysis revealed one major protein with an average molecular mass of 22 126.0 Da and some additional minor components. Electrospray LC-MS/MS of the enzymatically digested Norditropin sample showed deamidation of N(12)N(149) and N(159), oxidation of M(14), M(125) and M(170) and one amino acid exchange V(14) for M(14) present in <1% of Norditropin. While deamidation and oxidation may be due to technical reasons, the single amino acid exchange may reflect infidelity of translation rather than codon usage and copy editing by E. coli. PMID- 16372271 TI - Analysis of the Pasteurella multocida outer membrane sub-proteome and its response to the in vivo environment of the natural host. AB - This study describes the identification of outer membrane proteins (OMPs) of the bacterial pathogen Pasteurella multocida and an analysis of how the expression of these proteins changes during infection of the natural host. We analysed the sarcosine-insoluble membrane fractions, which are highly enriched for OMPs, from bacteria grown under a range of conditions. Initially, the OMP-containing fractions were resolved by 2-DE and the proteins identified by MALDI-TOF MS. In addition, the OMP-containing fractions were separated by 1-D SDS-PAGE and protein identifications were made using nano LC MS/MS. Using these two methods a total of 35 proteins was identified from samples obtained from organisms grown in rich culture medium. Six of the proteins were identified only by 2-DE MALDI-TOF MS, whilst 17 proteins were identified only by 1-D LC MS/MS. We then analysed the OMPs from P. multocida which had been isolated from the bloodstream of infected chickens (a natural host) or grown in iron-depleted medium. Three proteins were found to be significantly up-regulated during growth in vivo and one of these (Pm0803) was also up-regulated during growth in iron-depleted medium. After bioinformatic analysis of the protein matches, it was predicted that over one third of the combined OMPs predicted by the bioinformatics sub-cellular localisation tools PSORTB and Proteome Analyst, had been identified during this study. This is the first comprehensive proteomic analysis of the P. multocida outer membrane and the first proteomic analysis of how a bacterial pathogen modifies its outer membrane proteome during infection. PMID- 16372272 TI - The expression of ketohexokinase is diminished in human clear cell type of renal cell carcinoma. AB - For identification and targeting of tumor-associated marker proteins, the proteome of clear cell type of renal cell carcinoma (RCC) and normal kidney tissues was analyzed by 2-DE. Ketohexokinase (also called fructokinase), which catalyzes the phosphorylation of fructose to fructose 1-phosphate, was identified by MALDI-TOF MS and found to be expressed at low rates in the renal tumor tissues. We found a decreased amount of ketohexokinase mRNA in RCC compared to that observed in the normal kidney tissues by Northern blot. The activity of ketohexokinase in 20 clear cell RCC specimens and the 20 corresponding normal kidneys was investigated, and its activity was shown to be approximately 1.4-fold lower in the RCC specimens than in the normal kidney. Ketohexokinase activity in tumor stage pT3 RCC was 1.5-fold lower than in pT1 RCC. The level of ketohexokinase activity in histological grade 3 RCC was 1.8-fold lower than that in grade 1 cancer. In addition, using in situ hybridization, it was revealed that ketohexokinase in the normal kidney tissue was confined to the proximal tubular epithelial cells, while the expression of ketohexokinase in RCC tissues was extremely low. Our research results show that the expression of human ketohexokinase was diminished in clear cell RCC. PMID- 16372273 TI - Proteomic evaluation of cadmium toxicity on the midge Chironomus riparius Meigen larvae. AB - Heavy-metal pollution of aquatic ecosystems is a widespread phenomenon after industrial consumption. Whether aquatic organisms are adapted to the heavy-metal pollutants or not, such environmental stress causes changes in physiological responses. In this study, the aquatic midge, Chironomus riparius Meigen, was used to find changes of expression of proteins in relation to cadmium exposure. Dose response relationships between cadmium concentrations and mortality of 3rd instar midge larvae were observed and the protein levels were compared using PD-Quest after 2-DE. Comparing the intensity of protein spots, 21 proteins decreased and 18 proteins increased in response to cadmium treatment. With increased proteins, three enzymes such as S-adenosylmethionine decarboxylase, O-methyltransferase, and aspartokinase were involved in the glutathione biosynthesis and a key enzyme regulating fatty acid biosynthesis, oleyl-acyl carrier protein thioesterase was also identified. According to the functional classification of decreased levels of proteins, they were involved in energy production, protein fate, nucleotide biosynthesis, cell division, transport and binding, signal transduction, and fatty acid and phospholipid metabolism in the cell. In addition, phenol hydroxylase, thioesterase, zinc metalloprotease, and aspartate kinase were newly expressed after cadmium exposure at the concentration of the LC(10 )value. Therefore, these proteins seem to be potential biomarkers for cadmium exposure in the aquatic ecosystems. PMID- 16372274 TI - Establishment of a 2-D human urinary proteomic map in IgA nephropathy. AB - Immunoglobulin A nephropathy (IgAN) is the most common form of immune complex mediated glomerulonephritis worldwide. Although chronic renal failure develops in considerable numbers of IgAN patients, the exact etiology has not yet been clearly elucidated. To establish the urinary protein map of IgAN, we performed a urinary proteomic analysis. Thirteen patients with IgAN and 12 normal controls were recruited. Morning midstream spot urine samples were used with Centriprep ultrafiltration for concentration and desalting. 2-DE was performed and compared between IgAN and normal control, and urinary proteins were identified by MALDI TOF MS. A large number of protein spots were identified in IgAN and normal control samples, with means of 311 spots and 174 spots, respectively. Approximately 216 protein spots were detected as differentially expressed in IgAN. Among these, 82 spots were over-expressed, and 134 spots were under expressed compared to normal controls. A total of 84 differentially expressed spots, representing 59 different proteins, were finally identified in IgAN. We have established a urinary proteomic map of IgAN and this result helps in the identification. Further study is needed to determine the potential pathogenic role of these proteins. PMID- 16372275 TI - Alignment and statistical difference analysis of complex peptide data sets generated by multidimensional LC-MS. AB - A method for high-resolution proteomics analyses of complex protein mixtures is presented using multidimensional HPLC coupled to MS (MDLC-MS). The method was applied to identify proteins that are differentially expressed during fruit ripening of tomato. Protein extracts from red and green tomato fruits were digested by trypsin. The resulting highly complex peptide mixtures were separated by strong cation exchange chromatography (SCX), and subsequently analyzed by RP nano-LC coupled to quadrupole-TOF MS. For detailed quantitative comparison, triplicate RP-LC-MS runs were performed for each SCX fraction. The resulting data sets were analyzed using MetAlign software for noise and data reduction, multiple alignment and statistical variance analysis. For each RP-LC-MS chromatogram, up to 7000 mass components were detected. Peak intensity data were compared by multivariate and statistical analysis. This revealed a clear separation between the green and red tomato samples, and a clear separation of the different SCX fractions. MS/MS spectra were collected using the data-dependent acquisition mode from a selected set of differentially detected peptide masses, enabling the identification of proteins that were differentially expressed during ripening of tomato fruits. Our approach is a highly sensitive method to analyze proteins in complex mixtures without the need of isotope labeling. PMID- 16372276 TI - Identification of glycoconjugates in the urine of a patient with congenital disorder of glycosylation by high-resolution mass spectrometry. AB - More than 150 molecular species were detected in a single glycoconjugate fraction obtained from urine of a congenital disorders of glycosylation (CDG) patient by use of high-resolution FT-ICR MS. With respect to its high-mass accuracy and resolving power, FT-ICR MS represents an ideal tool for analysis of single components in complex glycoconjugate mixtures obtained from body fluids. The presence of overlapping nearly isobaric ionic species in glycoconjugate mixtures obtained from CDG patient's urine was postulated from fragmentation data of several precursor ions obtained by nanoESI Q-TOF CID. Their existence was confirmed by high-resolution/high-mass accuracy FT-ICR MS detection. High resolution FT-ICR mass spectra can, therefore, be generally considered for glycoscreening of complex mixture samples in a single stage. From the accurate molecular ion mass determinations the composition of glycoconjugate species can be identified. Particular enhancement of identification is offered by computer assisted calculations in combination with monosaccharide building block analysis, which can be extended by considerations of non-carbohydrate modifications, such as amino acids, phosphates and sulfates. Taking advantage of this strategy, the number of compositions assigned to mass peaks was significantly increased in a fraction obtained from urine by size exclusion and anion exchange chromatography. PMID- 16372277 TI - Protein sites of attack of N-chlorotaurine in Escherichia coli. AB - N-Chlorotaurine sodium (NCT) is a promising microbicidal agent for topical treatment of infections. Its targets of attack in Escherichia coli have been investigated by proteomics. Incubation in 1% NCT for 10 and 30 min revealed a change of the charge and a separation of numerous proteins into a series of spots with a different pI. Charge differences could be related to oxidation of cysteine residues to their corresponding sulfonic acids. Heat shock protein 60 appeared, while ribosome-releasing factor, d-ribose periplasmic binding protein, and malonyl-CoA transacylase spots decreased. These results indicate penetration of oxidation capacity into the bacteria and destruction of essential proteins by NCT. PMID- 16372278 TI - Sensitivity-enhanced 13C MR spectroscopy of the human brain at 3 Tesla. AB - A new coil design for sensitivity-enhanced 13C MR spectroscopy (MRS) of the human brain is presented. The design includes a quadrature transmit/receive head coil optimized for 13C MR sensitivity. Loss-less blocking circuits inside the coil conductors allow this coil to be used inside a homogeneous circularly polarized 1H B1 field for 1H decoupled 13C MRS. A quadrature 1H birdcage coil optimized for minimal local RF heating makes broadband 1H decoupling in the entire human brain possible at 3 Tesla while remaining well within international safety guidelines for RF absorption. Apart from a substantial increase in sensitivity compared to conventional small linear coils, the quadrature 13C coil combined with the quadrature 1H birdcage coil allows efficient cross polarization (CP) in the brain, resulting in an additional 3.5-fold sensitivity improvement compared to direct 13C measurements without nuclear Overhauser enhancement (NOE) or polarization transfer. Combined with the gain in power efficiency, this setup allows broadband 1H to 13C CP over large areas of the brain. Clear 13C resonances from glutamate (Glu), glutamine (Gln), aspartate (Asp), lactate (Lac), and gamma aminobutyrate (GABA) carbon spins in the human brain demonstrate the quality of 13C MR spectra obtained in vivo with this coil setup. PMID- 16372279 TI - Single acquisition water-fat separation: feasibility study for dynamic imaging. AB - Water-fat separation can be challenging in the presence of field inhomogeneities. Three-point (3-pt) water-fat separation methods achieve robust performance by measuring and compensating for field inhomogeneities; however, they triple the scan time. The "1+-pt" water-fat separation method proposed in this article for dynamic or repetitive imaging situations combines 3-pt methods' ability to correct for field inhomogeneities with the scan efficiency of a single acquisition method to achieve high temporal and spatial resolutions and robust water-fat separation. Single-echo data are collected with water and fat at a relative phase shift of an odd multiple of pi/2. To correct for undesired phase modulation, phase maps are estimated from a 3-pt calibration scan acquired prior to dynamic imaging. The phase maps are assumed to be slowly varying in time, so they may be used for correcting the phase of the subsequent single-echo signals at the same imaging location. Noise performance was investigated and shown to be equivalent to a single excitation acquisition. The 1+-pt method can also be used in conjunction with parallel imaging. In this situation, the calibration scans required by both methods can be integrated into a shared calibration scan. Promising results were obtained in breast, abdominal, and cardiac imaging applications. PMID- 16372280 TI - Inverse relation between in vivo amyloid imaging load and cerebrospinal fluid Abeta42 in humans. AB - OBJECTIVES: Amyloid-beta(42) (Abeta(42)) appears central to Alzheimer's disease (AD) pathogenesis and is a major component of amyloid plaques. Mean cerebrospinal fluid (CSF) Abeta(42) is decreased in dementia of the Alzheimer's type. This decrease may reflect plaques acting as an Abeta(42) "sink," hindering transport of soluble Abeta(42) between brain and CSF. We investigated this hypothesis. METHODS: We compared the in vivo brain amyloid load (via positron emission tomography imaging of the amyloid-binding agent, Pittsburgh Compound-B [PIB]) with CSF Abeta(42) and other measures (via enzyme-linked immunosorbent assay) in clinically characterized research subjects. RESULTS: Subjects fell into two nonoverlapping groups: those with positive PIB binding had the lowest CSF Abeta(42) level, and those with negative PIB binding had the highest CSF Abeta(42) level. No relation was observed between PIB binding and CSF Abeta(40), tau, phospho-tau(181), plasma Abeta(40), or plasma Abeta(42). Importantly, PIB binding and CSF Abeta(42) did not consistently correspond with clinical diagnosis; three cognitively normal subjects were PIB-positive with low CSF Abeta(42), suggesting the presence of amyloid in the absence of cognitive impairment (ie, preclinical AD). INTERPRETATION: These observations suggest that brain amyloid deposition results in low CSF Abeta(42), and that amyloid imaging and CSF Abeta(42) may potentially serve as antecedent biomarkers of (preclinical) AD. PMID- 16372281 TI - BOLD and CBV-weighted functional magnetic resonance imaging of the rat somatosensory system. AB - A multislice spin echo EPI sequence was used to obtain functional MR images of the entire rat brain with blood oxygenation level dependent (BOLD) and cerebral blood volume (CBV) contrast at 11.7 T. Maps of activation incidence were created by warping each image to the Paxinos rat brain atlas and marking the extent of the activated area. Incidence maps for BOLD and CBV were similar, but activation in draining veins was more prominent in the BOLD images than in the CBV images. Cerebellar activation was observed along the surface in BOLD images, but in deeper regions in the CBV images. Both effects may be explained by increased signal dropout and distortion in the EPI images after administration of the ferumoxtran-10 contrast agent for CBV fMRI. CBV-weighted incidence maps were also created for 10, 20, and 30 mg Fe/kg doses of ferumoxtran-10. The magnitude of the average percentage change during stimulation increased from 4.9% with the 10 mg Fe/kg dose to 8.7% with the 30-mg Fe/kg dose. Incidence of activation followed a similar trend. PMID- 16372282 TI - Emergence of human G9 rotavirus with an exceptionally high frequency in children admitted to hospital with diarrhea in Chiang Mai, Thailand. AB - Among 315 fecal specimens collected from children hospitalized with diarrhea in Chiang Mai, Thailand, in 2000-2001, group A rotavirus was detected in 107 (34.0%). Of these, 98 (91.6%) were G9, 6 (5.6%) were G3 and 3 (2.8%) were G2, respectively. Identification of their P-types demonstrated that 103 (96.3%) were P[8], 3 (2.8%) were P[4], and 1 (0.9%) was P[3] genotypes. Determination of G- and P-type combination revealed that all of G9 isolates were associated with P[8]. G9P[8] was the most predominant genotype and accounted for the majority (91.6%) of rotaviruses detected in this study. Molecular characterization of these G9 isolates demonstrated that all had long electropherotype, 96 of 98 (98.0%) belonged to subgroup II, one belonged to subgroup I and the other one was subgroup unidentifiable. All of G9 isolates possessed NSP4 genetic group B except for one isolate that showed dual genetic group specificities, B and C. The full length VP7 gene nucleotide sequences among 15 representatives of these G9 strains were found to be highly homologous with percent identities of 99.3%-100%. Comparison with other G9 strains recently isolated showed that their nucleotide sequences were closely related to those of the US strain, US1205 (98.7%-99.0%) and Thai strain, 97CM108 (98.1%-99.0%). Interestingly, they were most closely related to the Japanese strain, 00-SG2509VP7, isolated in the same epidemic season, with percent nucleotide sequence identity of 99.4%-99.8%. The data imply that G9 strains isolated in this study and a G9 strain isolated in Japan in the year 2000 might have descended from the same ancestor. PMID- 16372284 TI - Long-term monitoring of genotypic and phenotypic resistance to T20 in treated patients infected with HIV-1. AB - The aim of this study was to investigate the susceptibility to T20 and the dynamics of amino acid changes in HR1 and HR2 of gp41 of HIV-1 obtained from plasma, peripheral blood mononuclear cells (PBMC), and primary isolates (PI) in four highly antiretroviral-experienced patients. These patients received T20 plus an antiretroviral regimen and were followed-up over a period of 40-72 weeks. In one non-responder patient, N43D substitution was detected at 12 weeks of treatment, in association with a value of T20-IC50 of 10 microg/ml (10-fold increase). Double mutations N42T + N43D were observed in plasma RNA at 32 weeks and remained detectable up to 16 weeks after the withdrawal of the drug. The S138A substitution in HR2 was observed in plasma RNA at 32 weeks, and both in plasma RNA and in PI DNA at 40 weeks, associated with an increase of the T20-IC50 to 25 microg/ml (25-fold increase). Mutations V101G and E137K, not reported previously, were also observed in the HR2 region. Whether these new substitutions play a role in T20 resistance needs to be examined. In three temporary responders, coinciding with viral load rebound, G36D, and N42T substitutions were observed at 12, 24, and 40 weeks. G36D mutation was associated with a value of T20-IC50 of 5 microg/ml. The HR2 S138A mutation was detected after the detection of HR1 substitutions and was associated with an increase in the level of T20-IC50 to 125 microg/ml (125-fold increase) All these data reinforce the role of gp41 amino acids 36-45 and the potential influence of the HR2 S138A mutation in the genotypic/phenotypic resistance to T20. PMID- 16372285 TI - Protective efficacy, immunotherapeutic potential, and safety of hepatitis B vaccines. AB - Hepatitis B vaccines are highly effective and safe and have been incorporated into national immunization programs in over 150 countries. The major humoral immune response is to the common a determinant of the surface antigen protein of the virus. Approximately 5-10% of healthy immunocompetent subjects do not mount an antibody response (anti-HBs). Non-response is associated with different HLA-DR alleles and impaired Th cell response, among other factors such as route of injection, age, gender, body mass, and other factors. Important hepatitis B surface antigen variants have also been identified, which may have a potential impact on immunization and routine screening of blood, blood products and tissues, and organs for transplantation. Strategies for hepatitis B immunization are reviewed. Over 1,000 million doses of hepatitis B vaccine have been used with an outstanding record of safety. There is no evidence of an association between hepatitis B vaccines and the sudden infant death syndrome, chronic fatigue syndrome, and multiple sclerosis (MS). Several studies are in progress on treatment of chronic hepatitis B infection by immunization with multiple antigenic components, combination of vaccine with antiviral drugs and cytokines, T cell vaccines, DNA vaccines alone or with DNA encoded immunomodulatory cytokines, and direct genetic manipulation of antigen presenting cells. PMID- 16372283 TI - Soochong virus: an antigenically and genetically distinct hantavirus isolated from Apodemus peninsulae in Korea. AB - Hantaan (HTN) virus, the etiologic agent of clinically severe hemorrhagic fever with renal syndrome (HFRS), was first isolated in 1976 from lung tissue of a striped-field mouse (Apodemus agrarius) captured in Songnae-ri, Gyeonggi Province, Republic of Korea. Found primarily in mountainous areas, the Korean field mouse (A. peninsulae) is the second-most dominant field rodent species found throughout Korea. A new hantavirus, designated Soochong (SOO), was isolated in Vero E6 cells from four A. peninsulae captured in August 1997 at Mt. Gyebang in Hongcheon-gun, Mt. Gachil, Inje-gun, Gangwon Province, and in September 1998 at Mt. Deogyu, Muju-gun, Jeollabuk Province. The entire S, M, and L genomic segments of SOO virus, amplified by RT-PCR from lung tissues of seropositive A. peninsulae and from virus-infected Vero E6 cells, diverged from HTN virus (strain 76-118) by 15.6%, 22.8%, and 21.7% at the nucleotide level and 3.5%, 9.5%, and 4.6% at the amino acid level, respectively. Phylogenetic analyses of the nucleotide and deduced amino acid sequences, using the maximum parsimony and neighbor-joining methods, indicated that SOO virus was distinct from A. agrarius borne HTN virus. SOO virus shared a common ancestry with Amur virus from Far East Russia, as well as with H5 and B78 hantaviruses, previously isolated from HFRS patients in China. Cross-focus-reduction neutralizating antibody tests showed that SOO virus, which is the first hantavirus isolated in cell culture from A. peninsulae, could be classified as a new hantavirus serotype. PMID- 16372286 TI - Intrahepatic cytokine expression is downregulated during HCV/HIV co-infection. AB - HIV co-infection is associated with reduced HCV treatment response rates and accelerated HCV-related liver disease. Cytokines play an important role in regulating hepatic inflammation and fibrogenesis during chronic HCV infection, yet the roles of HIV and/or its therapies on cytokine expression are unknown. Total RNA was extracted from liver biopsies of 12 HCV mono-infected and 14 HCV/HIV co-infected persons. We used real-time PCR to quantify cytokines that contribute to innate and adaptive immune responses, including IFNalpha, IFNgamma, TNFalpha, TGFbeta(1), IL-2, IL-4, IL-8, IL-10, and IL-12p40. Positive- and negative-strand HCV RNA levels were quantified using a molecular beacon approach. Detection of positive-strand HCV RNA was 100% in both groups; negative-strand HCV RNA was detected in four (33%) HCV mono-infected persons and in nine (64%) HCV/HIV co-infected persons. Median strand-specific HCV RNA levels were not significantly different between the two groups. Detection rates of cytokine mRNAs were lower for the HCV/HIV co-infected group compared to the HCV mono-infected group; the detection rates for TNFalpha, IL-8, and IL-10 were statistically significant. Overall, cytokine mRNA quantities were lower for HCV/HIV co-infected compared to HCV mono-infected persons, with the exception of TGFbeta1. These data suggest that a defect in cytokine activation may occur in HCV/HIV co-infected persons that limits efficient clearance of HCV from the liver. PMID- 16372287 TI - Molecular detection and characterization of human enteroviruses directly from clinical samples using RT-PCR and DNA sequencing. AB - Enteroviruses are common human pathogens associated with a wide spectrum of symptoms ranging from asymptomatic infection to acute flaccid paralysis and neonatal multi-organ failure. Molecular methods that provide rapid diagnosis and increased sensitivity have been developed for the diagnosis of enterovirus infection using oligonucleotide primers complementary to conserved sequences located in the 5' untranslated region (UTR), but data generated from these regions are not sufficiently discriminatory for typing due to the lack of correlation between their nucleic acid sequence and serotype specificity. Sequences derived from the gene encoding the capsid VP1 correlate with serotype, and therefore provide the opportunity for the development of molecular typing methods consistent with present serogical methods. In this study, oligonucleotide primers that amplify a region of the 5'UTR to detect enterovirus RNA, and the region encoding the enterovirus VP1 N-terminus to characterize virus strains were used in nested and semi-nested RT-PCRs, respectively. The ability of the VP1 RT PCR to amplify diverse viruses within genotypes and genogroups was confirmed by the correct identification of both prototype strains, and strains circulating currently of the same genotypes. The molecular methods proved their utility through the detection of enteroviruses that failed to grow in cell culture, their subsequent characterization and the characterization of strains that failed to serotype in neutralization assays. Molecular methods increased significantly the sensitivity of detection (P < 0.001) and of characterization (P < 0.01) of enteroviruses when compared to classical methods. PMID- 16372288 TI - Molecular analysis of VP4, VP7, and NSP4 genes of P[6]G2 rotavirus genotype strains recovered from neonates admitted to hospital in Belem, Brazil. AB - This investigation describes the molecular characterization of P[6]G2 rotavirus strains from hospitalized neonates with community-acquired diarrhea (CAD), nosocomial diarrhea (ND), and asymptomatic nosocomial infection (ANI) in Belem, Brazil. Twenty-six rotavirus strains with P[6]G2 genotype were sequenced to genes coding for VP4, VP7, and NSP4 proteins. Phylogenetic analysis of the VP4 gene, including prototype strains RV3, ST3, M37, and U1205, showed that local P[6]G2 strains clustered forming a distinct lineage (bootstrap of 99%). Brazilian P[6]G2 strains had the highest homology (ranging from 96.0%-98.3%) with the African strain GR1107, G4P[6]. Phylogenetic tree for VP7 gene was constructed including old and new G2 African strains SA3958GR/97, SA356PT/96, SA514GR/87, SA4476PT/97, BF3676/99, GH1803/99, and representative strains of G1, G3, G4, G5, G8, and G9 genotypes. The Brazilian P[6]G2 samples fell into a distinct group (bootstrap value of 97%) and showed homology rates ranging from 92.1% to 93.5% with P[6]G2 African strains BF3676/99, GH1803/99, and SA3958GR/97. Nucleotide sequence analysis of the NSP4 gene, including human prototype strains S2, KUN, DS-1, RV5, RV3 and ST3, and animal prototype OSU, showed that all neonatal isolates fell into genotype A and clustered with a bootstrap value of 100%, with in-group similarities ranging from 99.3% to 100%. In this study no significant differences in nucleotide sequences of the VP4, VP7, and NSP4 genes could be observed when comparing diarrheic (CAD and ND) and non-diarrheic (ANI) babies. Monitoring of rotavirus strains in hospital environments is of particular importance, since it is claimed currently that an efficacious rotavirus vaccine, when available for routine use, will determine an impact on hospital-acquired rotavirus disease. PMID- 16372289 TI - Hepatitis C virus infection among dialysis patients in Tunisia: incidence and molecular evidence for nosocomial transmission. AB - In order to study the incidence of hepatitis C virus (HCV) infection in Tunisian haemodialysis patients and detect its nosocomial transmission, 395 patients were enrolled in a prospective study (November 2001-2003). HCV serological and virological status was determined initially using, respectively a third generation ELISA and an RT-PCR qualitative assay. The genotype of the HCV isolates was determined by sequencing NS5B region. The issue of nosocomial transmission was addressed by sequencing the HVR-1 region of the E2 gene. About 20% of the patients had anti-HCV antibodies and HCV-RNA was detected in 73% of the anti-HCV positive patients. Two cases of de novo HCV infection were identified in two dialysis centers, during virological follow-up of patients susceptible to HCV infection. The incidence of de novo HCV infection was 0.5%. Determining the genotypes in the first center disclosed that all HCV-positive patients were infected with genotype 1b; sequencing of the HVR-1 region of the E2 gene provided strong evidence that the isolate from the newly infected patient and another infected dialysis patient were closely related, confirming nosocomial contamination. The investigation of the second center is pending. Besides, one patient with negative HCV serology had detectable HCV-RNA at the beginning of the study. This case had HCV genotype 1b, two other infected dialysis patients in the same unit had HCV genotypes 4k and 3a; thus precluding nosocomial transmission. Thanks to molecular and phylogenetic methods, one case of nosocomial HCV transmission in haemodialysis was confirmed. Epidemiological investigation suggested nosocomial transmission via the medical and/or nursing staff. PMID- 16372290 TI - A variable region in the Crimean-Congo hemorrhagic fever virus L segment distinguishes between strains isolated from different geographic regions. AB - Alignment of Crimean-Congo hemorrhagic fever virus (CCHFV) L genome segment full length sequences reveals an overall high level of conservation among strains, with greater than 90% of translated amino acid residues strictly conserved. However, a region of marked variability identified previously, corresponding to L polyprotein amino acid positions 760-810, shares only 40% overall identity between strains. The variable regions sequences of 16 laboratory-adapted CCHFV strains were determined, including 11 strains from European Russia, one strain from Bulgaria, and four strains from the Central Asian countries of Tajikistan, Turkmenistan, and Uzbekistan. Phylogenetic analysis demonstrates this L segment variable region sequence divides CCHFV strains into similar geographically defined groupings observed for S segment-derived trees, but with higher bootstrap support and a much smaller character set required for analysis. PMID- 16372291 TI - Rotavirus surveillance in the city of Rio de Janeiro-Brazil during 2000-2004: detection of unusual strains with G8P[4] or G10P[9] specificities. AB - Rotavirus diarrhea is a potentially life-threatening disease that affects millions of children annually around the world. Because protection against rotavirus disease is thought to be type specific, continuous rotavirus surveillance before and after implementation of a vaccine is still of essential importance. Rotavirus surveillance has been conducted in the city of Rio de Janeiro, Brazil since 1982. In the present study, we report rotavirus surveillance data in Rio de Janeiro city from 2000 to 2004. One hundred twenty nine of 1,568 (8.2%) stool samples, collected from children with acute diarrhea between January 2000 and July 2004 were rotavirus-positive. One hundred twenty eight of the 129 (99.2%) rotavirus-positive samples were genotyped for G and/or P specificity. G1 was the predominant strain (49.6%, 64/129) followed by G9 (30.2%, 39/129), and G4 (17.8%, 23/129); G2 and G3 viruses were not detected. One sample (0.8%) was non-typeable. P genotypes were determined for 124 of the 129 (96%) samples, and P[8] was the predominant genotype (90.6%, 117/129). Genotypes P[4] and P[9] were detected in two (1.6%) samples each; one (0.8%) sample presented P[6] genotype; and five (3.8%) samples were non-typeable. Two samples (1.6%) presented mixed P genotypes (P[6] + P[8]). Two unusual strains were isolated: a G8P[4] strain isolated from a non-hospitalized child with diarrhea and a G10P[9] strain isolated from a hospitalized child with diarrhea. PMID- 16372292 TI - Distribution and genetic analysis of TTV and TTMV major phylogenetic groups in French blood donors. AB - TTV and TTMV (recently assigned to the floating genus Anellovirus) infect human populations (including healthy individuals) at high prevalence (>80%). They display notably high levels of genetic diversity, but very little is known regarding the distribution of Anellovirus genetic groups in human populations. We analyzed the distribution of the major genetic groups of TTV and TTMV in healthy voluntary blood donors using group-independent and group-specific PCR amplifications systems, combined with sequence determination and phylogenetic analysis. Analysis of Anellovirus groups revealed a non-random pattern of group distribution with a predominant prevalence of TTV phylogenetic groups 1, 3, and 5, and of TTMV group 1. Multiple co-infections were observed. In addition, TTMV sequences exhibiting a high genetic divergence with reference sequences were identified. This study provided the first picture of the genetic distribution of the major phylogenetic groups of members of the genus Anellovirus in a cohort of French voluntary blood donors. Obtaining such data from a reference population comprising healthy individuals was an essential step that will allow the subsequent comparative analysis of cohorts including patients with well characterized diseases, in order to identify any possible relationship between Anellovirus infection and human diseases. PMID- 16372293 TI - Accuracy of HCV-RNA PCR tests for diagnosis or exclusion of vertically acquired HCV infection. AB - The aim of the study was to estimate the sensitivity, specificity, positive (PPV) and negative (NPV) predictive values, and likelihood ratios for HCV-RNA PCR tests for the early diagnosis or exclusion of HCV infection in vertically exposed children. Data were included for children with confirmed HCV infection status from a European multi-center study. Confirmation was dependent on antibody status at or beyond 18 months, the 'gold standard' measure of infection status against which the use of qualitative HCV-RNA PCR tests was assessed. Of the 547 children included in this analysis, 193 were HCV-infected and 354 were not. Sensitivity of the HCV-RNA PCR test was low at birth (22%), but increased to 85% by 6 months. Specificity of RNA PCR was constant over age at 98%. The PPV of the PCR test rose from 33% at birth to 78% at 9 months of age, while NPV ranged from 96% to 99%. The high positive likelihood ratios from 1 month of age indicate strong evidence to diagnose infection but the negative likelihood ratios were consistent with weak evidence to exclude infection. The results suggest that the first qualitative HCV-RNA PCR test should be delayed until after the first month of life given the low sensitivity in the first few weeks. Although a negative test result after this time indicates probable absence of infection, this should be confirmed with a negative anti-HCV antibody test between 9 and 15 months of age as negative PCR results can be observed in infected children with fluctuations in viremia. PMID- 16372294 TI - Reduction of mother-to-child transmission of HIV at Saint Camille Medical Centre in Burkina Faso. AB - One thousand three hundred and twenty-eight pregnant women with less than 32 weeks of amenorrhea received voluntary counseling and testing at Saint Camille Medical Center from May 1, 2002 to December 30, 2004. Following informed consent and pre-test counseling, HIV screening was performed in 1,202 women. According to the prevention protocol, HIV-positive women received a single dose of Nevirapine (200 mg) during their labor, while their newborn received a single dose of Nevirapine (2 mg/kg) within 72 hr from birth. HIV seroprevalence (11.2%) was higher than in the overall population. One hundred and ninety-three children were born at the end of December 2004; 53 children (27.5%) followed a short breastfeeding protocol for 4 months, while 140 (72.5%) were fed artificially. All the children underwent RT-PCR test for HIV 5-6 months after their birth: 173 (89.6%) were HIV negative whilst 20 children (10.4%) were HIV positive. Out of the 20 positive children 5/53 (9.4%) had received breast milk for 4 months, while the remaining 15/140 (10.7%) had been fed artificially (P = 0.814). Artificially fed babies (3/140 (2.1%)) and 1/53 (1.9%) of those breast fed for 4 months deceased according to mortality rate of HIV-positive children. This shows that there is no statistically significant difference (P = 0.648) between the mortality of artificially fed (3/140 or 2.1%) and breast-fed (1/53 or 1.9%) children. Artificially fed children (20/140 (14.3%)) and 5/53 (9.4%) of breast fed children died within 6-10 months. This figure indicates that there is no significant difference between the mortality rate of artificially and that of breast-fed children (P = 0.427). Although the HIV prevention program reduced significantly the vertical transmission of HIV at Saint Camille Medical Center, the mortality of artificially fed children was still high due to gastrointestinal diseases. The HIV diagnosis by RT-PCR technique was of great help in the early identification of HIV-infected children. PMID- 16372295 TI - Reverse transcriptase activity for quantitation of HIV-1 subtype C in plasma: relation to RNA copy number and CD4 T-cell count. AB - The present study monitored the changes in human immunodeficiency virus (HIV) viral load using a reverse transcriptase (RT) assay and an HIV-1 RNA based assay, and relates these data to the dynamics of CD4 cell counts. The samples examined originate from a prospective study of HIV-1 subtype C infected, untreated Ethiopians followed twice yearly over a period of up to 5 years. The ExaVir Load test, version 1, was used for isolation and quantitation of HIV-1 RT in plasma. The RT activities recovered were compared to the HIV-1 RNA copy numbers, which had been determined previously by the NucliSens HIV-1 QT Test. There was a significant correlation between the data obtained in the two tests (r = 0.65, P < 0.0001). During follow-up, the median RT and RNA levels increased more or less in parallel up to approximately four times the values at admittance. CD4 cell counts, which had also been determined previously, decreased slowly but continuously from approximately 310 to 190 CD4 cells/ml. In the majority of individual patients, there was an inverse correlation between CD4 T-cell counts and RT activity, and with the RNA copy number, and the data obtained by either test could be used to predict CD4 T-cell counts. The ExaVir Load test thus provides data equivalent to the estimation of the number of HIV-1 RNA copies for the prediction of CD4 T-cell counts. It is based on a simple technique, can be run in any routine diagnostic laboratory, and is a competitive alternative for use in resource limited settings. PMID- 16372296 TI - Characteristics of hepatitis B virus in Ghana: full length genome sequences indicate the endemicity of genotype E in West Africa. AB - Hepatitis B virus (HBV) genotypes from A to H have distinct geographical distributions and have been shown to affect the clinical features as well as the course of the HBV infection. HBV genotype E has been found only in Africa. However, the complete genomes of this genotype, which were isolated mainly from West Africa, were available only for a few samples. In this study, five full length genomes and seven other small S genes of HBV strains from Ghanaian blood donors were sequenced and investigated. Following phylogenetic analysis, all of the Ghanaian HBV strains were clustered closely in genotype E. All of the 12 small S genes showed the same characteristic of subtype ayw4. The complete genomes of the five Ghanaian strains showed marked similarity with each other and with the reported genotype E strains (96.7%-99.1%). Genotype E strains showed low intra-genotypic diversity (1.8%) and carried the conserved signature pattern in pre-S1 as well as in the full genome sequence. Of note, the finding of the G145R escape mutant in an unvaccinated Ghanaian blood donor might raise concern as to the ongoing nation-wide hepatitis B vaccination program in Ghana. PMID- 16372297 TI - Transmission in vitro of hepatitis C virus from persistently infected human B cells to hepatoma cells by cell-to-cell contact. AB - Virus cell-to-cell spread has been reported for many different viruses and may contribute to pathogenesis of viral disease. The role played by cell-to-cell contact in hepatitis C virus (HCV) transmission was studied in vitro by cell co cultivation experiments. A human lymphoblastoid B-cell line, infected persistently with HCV in vitro (TO.FE(HCV)), was used as HCV donor [Serafino et al., 2003]; recipient cells were the human hepatoma HepG2 cell line. Both cell types were co-cultured for 48 hr to allow the cell-to-cell contacts. The hepatoma HepG2 cells are not permissive to free-virus infection, but they were infected successfully using TO.FE(HCV) cells as source of virus. The kinetics of viral RNA synthesis and the percentage of infected cells were compared in cell-mediated-and cell-free-viral infection. After co-cultivation, a consistent proportion of hepatoma cells replicated HCV and stably expressed viral antigens. Virus produced was infectious as demonstrated by the ability to reinfect fresh B-cells. This cell model shows that permissiveness to HCV infection can be achieved in vitro in non-permissive hepatoma cells by direct cell-to-cell contacts with infected human B-cells. This mechanism of virus spread may also play a pathogenic role in vivo. PMID- 16372298 TI - Assessment of early virological response to antiviral therapy by comparing four assays for HCV RNA quantitation using the international unit standard: implications for clinical management of patients with chronic hepatitis C virus infection. AB - WHO International Standards for nucleic acid tests are used widely to compare the different assays used in HCV RNA quantitation. The aim of the study was to assess the impact of the international unit standard for measuring HCV RNA in the management of patients with chronic hepatitis C virus (HCV) infection. Twenty seven naive patients infected chronically by HCV were treated with ribavirin plus PEG-interferon-alfa-2b for 48 weeks. SVR was obtained for 16 patients (the other were non-responders). For HCV RNA quantitation, four assays were undertaken: Versant HCV RNA 3.0 (Bayer), Real time PCR (TaqMan, Roche), LCx HCV RNA (Abbott), and Cobas Amplicor-Monitor v2 (Roche). Considering a 2-log decline at Week 12 after the beginning of therapy, discordant results were found with the four HCV RNA methods in predicting SVR or non-response. At Week 4 and Week 12, significant differences were observed between Versant HCV RNA 3.0 versus PCR HCV Taqman, Versant HCV RNA 3.0 versus LCx HCV RNA, Cobas Monitor Amplicor HCV 2.0 versus LCx HCV RNA, and Cobas Monitor Amplicor HCV 2.0 versus PCR HCV Taqman (P < 0.001). The HCV RNA cutoff, given a 100% negative predictive value at Week 4 and Week 12, differed with the assays used to quantify HCV RNA, despite the use of the IU/ml units. Eighty-nine percent of serum values for HCV RNA were concordant by the IU standard. All assays, however, failed to detect HCV RNA in some cases. Despite the use of the IU standard HCV-infected patients might be monitored with only one assay. PMID- 16372299 TI - Type I interferon inhibits Crimean-Congo hemorrhagic fever virus in human target cells. AB - Crimean-Congo hemorrhagic fever virus (CCHFV) is a causative agent of severe hemorrhagic fever occurring sporadically in parts of Africa, Asia, Southeast Europe, and the Middle East. Its recent recognition as a potential agent of bioterrorism/biowarfare highlights the need for effective antiviral therapy. In this study, it is shown that human endothelial cells are permissive to CCHFV. It is also shown that interferon-alpha inhibits the growth of CCHFV in human endothelial and hepatoma cells, reducing virus yields by a factor of 100-1,000. By using a siRNA approach, it was demonstrated that the interferon-induced MxA GTPase is a major factor mediating the antiviral effect against CCHFV, in agreement with previous findings showing that recombinant MxA inhibits CCHFV replication by interacting with the viral nucleocapsid protein. The identification of intrinsic cellular resistance factors that block CCHFV replication may help in designing novel antiviral agents. PMID- 16372300 TI - Repeated HIV-1 resistance genotyping external quality assessments improve virology laboratory performance. AB - The performance of French virology laboratories belonging to the ANRS network has been assessed annually for 3 years. The performance of these laboratories was compared between the years 2002 and 2003. Ten and 7 coded samples were sent to 38 virology laboratories in 2002 and 45 virology laboratories in 2003, respectively. Each panel of coded samples included at least one HIV-negative control, a pair of duplicate specimens, samples with a wide range of viral loads, and samples with a large number of resistance mutations. The laboratories used their standard sequencing procedures and were asked to report the amino acids at codons associated with resistance mutations, based on the IAS-USA expert panel list. The reference amino acid sequences were defined as those most frequently reported by the participants. The specificity of detection of RT mutations was significantly better in 2003 (99.9%) than in 2002 (99.7%) (P = 0.05). There was no difference between 2002 and 2003 in the specificity of detection of protease mutations (99.6% and 99.8%) or the sensitivity of detection of RT mutations (98.8% and 98.2%). The sensitivity of detection of protease mutations improved significantly between 2002 and 2003 (97.6% and 99.0%, respectively; P = 0.037). The proportion of laboratories reporting fully accurate results, in terms of amplification, specificity, sensitivity, and reproducibility, tended to increase between 2002 and 2003 (P = 0.077). No errors were made by 19% of laboratories in 2002, compared to 42% in 2003. These results show the value of repeated external quality assessments. PMID- 16372301 TI - Dengue virus infection of human microvascular endothelial cells from different vascular beds promotes both common and specific functional changes. AB - Dengue shock syndrome (DSS), the major life threatening outcome of severe dengue disease, which occurs in some patients in the course of dengue infection, is the consequence of plasma leakage in the microvascular territories. Data from clinical and in vitro studies suggest that an inadequate immunological response is partly responsible for the pathophysiology of DSS, but few is known concerning the consequences of direct infection of endothelial cells by dengue virus per se. In this study, an attempt was made to study the response of two microvascular human cell lines originating, respectively, from liver and dermis to infection by a dengue type 2 virus, by analyzing the virus-induced modulation of functional markers. It is shown that the two microvascular cell lines exhibit both common and specific behaviors upon infection. In particular, LSEC and HMEC-1 replicate efficiently the low-passage virus and respond to infection by over-producing inflammatory mediators involved in the cross talk with circulating immune cells. However, direct infection modulates differently the cell surface expression of molecules critically involved in the interactions between endothelial and inflammatory cells. ICAM-1 and HLA-I are up regulated as a consequence of infection in LSEC whereas direct infection results in downregulation of ICAM-1 in HMEC-1. The present results show that infection of human microvascular cells by unadapted dengue virus results in both common and specific activation patterns depending likely on the tissue origin of the cells, thus suggesting that endothelia from different territories may contribute differently to the pathophysiological events in the course of dengue infection. PMID- 16372302 TI - Evolution of EV71 genogroup in Taiwan from 1998 to 2005: an emerging of subgenogroup C4 of EV71. AB - In Taiwan, enterovirus 71 (EV71) has played an important role in severe enterovirus-related cases every year since the devastating outbreak in 1998. Three genogroups A, B, C occur worldwide; with the B and C genogroups being subdivided into B1-B4 and C1-C4 subgenogroups respectively. To understand the mutation of the EV71 genogroup in Taiwan before and after 1998, a total of 54 worldwide strains were studied including 41 Taiwanese strains obtained in 1986 and 1998-2004. A fragment of 207 bp of the VP4 region was amplified and sequenced. Genetic analysis was performed using MEGA software (version 3.0) for the nucleotide sequence alignment and phylogenetic analysis. In Taiwan, the subgenogroup B1 was predominant before 1998 while subgenogroup C2 was the major etiologic group in 1998 outbreak. A minor etiologic group outbreak in 1998, subgenogroup B4, became predominant during the period from 1999 to 2003. In this study, subgenogroup C4 emerged and became predominant in 2004 in Taiwan. The nucleotide differences between B1 and C2, C2 and B4, B4 and C4 were 20%-26%, 19% 27%, 18%-22%, respectively. Nucleotide sequence alignment revealed 67 substitutions. Most of the substitutions (62/67) were silent mutations. This is the first report about the emergence of EV71 subgenogroup C4 in Taiwan. PMID- 16372303 TI - Conformation analysis of aspartame-based sweeteners by NMR spectroscopy, molecular dynamics simulations, and X-ray diffraction studies. AB - We report here the synthesis and the conformation analysis by 1H NMR spectroscopy and computer simulations of six potent sweet molecules, N-[3-(3-hydroxy-4 methoxyphenyl)-3-methylbutyl]-alpha-L-aspartyl-S-tert-butyl-L-cysteine 1 methylester (1; 70 000 times more potent than sucrose), N-[3-(3-hydroxy-4 methoxyphenyl)-3-methylbutyl]-alpha-L-aspartyl-beta-cyclohexyl-L-alanine 1 methylester (2; 50 000 times more potent than sucrose), N-[3-(3-hydroxy-4 methoxyphenyl)-3-methylbutyl]-alpha-L-aspartyl-4-cyan-L-phenylalanine 1 methylester (3; 2 000 times more potent than sucrose), N-[3,3-dimethylbutyl] alpha-L-aspartyl-(1R,2S,4S)-1-methyl-2-hydroxy-4-phenylhexylamide (4; 5500 times more potent than sucrose), N-[3-(3-hydroxy-4-methoxyphenyl)propyl]-alpha-L aspartyl-(1R,2S,4S)-1-methyl-2-hydroxy-4-phenylhexylamide (5; 15 000 times more potent than sucrose), and N-[3-(3-hydroxy-4-methoxyphenyl)-3-methylbutyl]-alpha-L aspartyl-(1R,2S,4S)-1-methyl-2-hydroxy-4-phenylhexylamide (6; 15 000 times more potent than sucrose). The "L-shaped" structure, which we believe to be responsible for sweet taste, is accessible to all six molecules in solution. This structure is characterized by a zwitterionic ring formed by the AH- and B containing moieties located along the +y axis and by the hydrophobic group X pointing into the +x axis. Extended conformations with the AH- and B-containing moieties along the +y axis and the hydrophobic group X pointing into the -y axis were observed for all six sweeteners. For compound 5, the crystal-state conformation was also determined by an X-ray diffraction study. The result indicates that compound 5 adopts an L-shaped structure even in the crystalline state. The extraordinary potency of the N-arylalkylated or N-alkylated compounds 1-6, as compared with that of the unsubstituted aspartame-based sweet taste ligands, can be explained by the effect of a second hydrophobic binding domain in addition to interactions arising from the L-shaped structure. In our examination of the unexplored D zone of the Tinti-Nofre model, we discovered a sweet-potency enhancing effect of arylalkyl substitution on dipeptide ligands, which reveals the importance of hydrophobic (aromatic)-hydrophobic (aromatic) interactions in maintaining high potency. PMID- 16372304 TI - Effect of pH and ionic strength on the physical stability of adenovirus type 5. AB - The thermal stability of adenovirus type 5 (Ad5) was investigated over the pH range 3-8 employing a variety of biophysical techniques under conditions of low and high ionic strength. Analysis of the structural stability of Ad5 by dynamic light scattering, intrinsic and extrinsic fluorescence, and second derivative UV absorption spectroscopies suggest that the capsid stability of Ad5 increases with decreasing pH under both ionic strength conditions. Significant aggregation, however, was observed at pH < or = 5 under conditions of low ionic strength. These studies also suggest that the physical stability of Ad5 is significantly enhanced under acidic conditions in the presence of 1 M NaCl. Evaluation of the quaternary structural stability of Ad5 by dynamic light scattering and extrinsic fluorescence spectroscopy suggest that the Ad5 capsid undergoes a two-step dismantling process wherein the viral particles initially expand in size near 50 degrees C and the DNA core is at least partially exposed to the surrounding solvent. Complete capsid disassembly and total exposure of the DNA core follows at higher temperatures. Data generated during these studies were combined employing a multidimensional eigenvector approach that combines data from numerous techniques into a colored representation. This picture, or "empirical phase diagram," provides an intuitive representation of the physical stability of Ad5 over the pH range 4-8 from 10 degrees C to 85 degrees C. PMID- 16372305 TI - Microscopic image analysis techniques for the morphological characterization of pharmaceutical particles: Influence of process variables. AB - This study investigated the effects of various process variables on some of the shape factors most widely used in the morphological characterization of pharmaceutical particles (including circularity, aspect ratio, er, and the recently proposed Vr and Vp). After optimization of the illumination system and greyscale thresholds for discrimination of particle background, we evaluated the effects of process variables within the image capture and analysis system, including the image capture device (video or digital camera), image color information (24-bits-per-pixel RGB or 8-bits-per-pixel black and white), file type (JPG or TIFF), and JPG file compression ratio. A key aspect was evaluation of the effect of scaling factor (microm/pixel), dependent on real pixel size and optical magnification, on shape factor estimates. The results obtained indicate that accurate estimation of shape factors requires use of a scaling factor below a certain maximum; however, use of very low scaling factors will mean that the field of view is very small, so that it will contain very few particles, implying a time-consuming increase in the number of images that must be analyzed. Finally, we use statistical procedures to estimate the minimum number of particles that must be analyzed in order to provide accurate estimates of shape factors. PMID- 16372306 TI - Phase solubility and inclusion complex of itraconazole with beta-cyclodextrin using supercritical carbon dioxide. AB - Phase-solubility techniques were used to assess the formation of inclusion complex between itraconazole and beta-cyclodextrin. The stability constant and free energies of transfer of itraconazole from aqueous solution to the cavity of beta-cyclodextrin were calculated. Itraconazole solubility in supercritical carbon dioxide (SC CO(2)) was measured at different temperatures and pressures. Drug formulations of itraconazole were prepared by complexation of the drug into beta-cyclodextrin using SC CO(2). Effects of temperature and pressure on inclusion yield of the prepared complexes were studied. The solvent-free inclusion complexes obtained from this method were characterized by UV spectroscopy, differential scanning calorimetry, powder X-ray diffraction, and scanning electron microscopy and compared to those obtained from physical mixing and coprecipitation methods. Results showed that beta-cyclodextrin significantly improved solubility of itraconazole in aqueous solutions. The free energies of transfer of itraconazole from aqueous solution to the cavity of beta-cyclodextrin increased negatively with increasing beta-cyclodextrin concentration. Higher inclusion yields were obtained in the SC CO(2) method compared to physical mixing and coprecipitation methods. Both temperature and pressure had significant effects on itraconazole solubility in SC CO(2) and the inclusion yield of the complex prepared by SC CO(2) method. PMID- 16372307 TI - Measuring biomolecular binding events with a compact disc player device. PMID- 16372308 TI - Multivariate statistical tools applied to the characterization of the proteomic profiles of two human lymphoma cell lines by two-dimensional gel electrophoresis. AB - Mantle cell lymphoma (MCL) cell lines have been difficult to generate, since only few have been described so far and even fewer have been thoroughly characterized. Among them, there is only one cell line, called GRANTA-519, which is well established and universally adopted for most lymphoma studies. We succeeded in establishing a new MCL cell line, called MAVER-1, from a leukemic MCL, and performed a thorough phenotypical, cytogenetical and molecular characterization of the cell line. In the present report, the phenotypic expression of GRANTA-519 and MAVER-1 cell lines has been compared and evaluated by a proteomic approach, exploiting 2-D map analysis. By univariate statistical analysis (Student's t test, as commonly used in most commercial software packages), most of the protein spots were found to be identical between the two cell lines. Thirty spots were found to be unique for the GRANTA-519, whereas another 11 polypeptides appeared to be expressed only by the MAVER-1 cell line. A number of these spots could be identified by MS. These data were confirmed and expanded by multivariate statistical tools (principal component analysis and soft-independent model of class analogy) that allowed identification of a larger number of differently expressed spots. Multivariate statistical tools have the advantage of reducing the risk of false positives and of identifying spots that are significantly altered in terms of correlated expression rather than absolute expression values. It is thus suggested that, in future work in differential proteomic profiling, both univariate and multivariate statistical tools should be adopted. PMID- 16372311 TI - Bi-3H-diazirin-3-yls as precursors of highly strained cycloalkynes. PMID- 16372313 TI - Detailed methylation analysis of CpG islands on human chromosome region 9p21. AB - Deletion of 9p21 is the most commonly reported chromosomal abnormality in pediatric acute lymphoblastic leukemia, and published data suggest that the maternal chromosome is preferentially deleted. Preferential maternal deletion of 9p21 and reports of a differentially methylated region (DMR) and of parental effects in mice with lymphoma suggest there may be an unrecognized imprinted locus in this region. To screen for DMRs, we used the mcrBC/HpaII screening method and peripheral-blood DNA. Of 36 CpG islands within an 8.5-Mb region of 9p21, seven were identified as putative DMRs and were further analyzed by bisulfite sequencing. Neither any of the CpG islands nor a previously published putative DMR nearby showed evidence of differential parental methylation; however, the published DMR did demonstrate sequence-dependent differential methylation. Our data, which showed heterogeneous and low-level methylation of CpG islands, have obvious implications for methylation studies. PMID- 16372315 TI - Molecular mobility of nifedipine-PVP and phenobarbital-PVP solid dispersions as measured by 13C-NMR spin-lattice relaxation time. AB - Amorphous nifedipine-PVP and phenobarbital-PVP solid dispersions with various drug contents were prepared by melting and subsequent rapid cooling of mixtures of PVP and nifedipine, or phenobarbital. Chemical shifts and spin-lattice relaxation times (T(1)) of PVP, nifedipine, and phenobarbital carbons were determined by (13)C-CP/MAS NMR to elucidate drug-PVP interactions and the localized molecular mobility of drug and PVP in the solid dispersions. The chemical shift of the PVP carbonyl carbon increased as the drug content increased, appearing to reach a plateau at a molar ratio of drug to PVP monomer unit of approximately 1:1, suggesting hydrogen bond interactions between the PVP carbonyl group and the drugs. T(1) of the PVP carbonyl carbon in the solid dispersions increased as the drug content increased, indicating that the mobility of the PVP carbonyl carbon was decreased by hydrogen bond interactions. T(1) of the drug carbons increased as the PVP content increased, and this increase in T(1) became less obvious when the molar ratio of PVP monomer unit to drug exceeded approximately 1:1. These results suggest that the localized motion of the PVP pyrrolidone ring and the drug molecules is reduced by hydrogen bond interactions. Decreases in localized mobility appear to be one of the factors that stabilize the amorphous state of drugs. PMID- 16372314 TI - Influence of aggregation on immunogenicity of recombinant human Factor VIII in hemophilia A mice. AB - Recombinant human factor VIII (rFVIII), a multidomain glycoprotein is used in replacement therapy for treatment of hemophilia A. Unfortunately, 15%-30% of the treated patients develop inhibitory antibodies. The pathogenesis of antibody development is not completely understood. The presence of aggregated protein in formulations is generally believed to enhance the immune response. rFVIII has a tendency to aggregate but the effect of such aggregation on the immunogenicity of rFVIII is not known. We have, therefore, characterized aggregated rFVIII produced by thermal stress and evaluated its effect on the immunogenicity of rFVIII in hemophilia A mice. Aggregated rFVIII alone and mixtures of rFVIII with aggregated rFVIII were less immunogenic than native rFVIII. In vitro Th-cell proliferation studies and cytokine analyses conducted on splenocytes obtained from immunized animals suggest that aggregated rFVIII behaves as a unique antigen compared to native monomeric rFVIII. The antigenic properties of the aggregated and native rFVIII were compared using ELISAs (epitope availability) and cathepsin-B (an antigen processing enzyme) digestion. The data suggest significant differences in the antigenic properties of rFVIII and aggregated rFVIII. Overall it appears that aggregated rFVIII does not enhance the immunogenicity (inhibitor development) of rFVIII in hemophilia A mice but rather acts as a distinct antigen. PMID- 16372316 TI - Systemic oxybutynin decreases afferent activity of the pelvic nerve of the rat: new insights into the working mechanism of antimuscarinics. AB - AIMS: In a rat model, intravesical oxybutynin was recently shown to suppress pelvic afferent nerves. This study evaluates if a similar effect exists after systemic administration of oxybutynin. METHODS: Twenty-four single afferent bladder nerves were identified in 15 rats. Based on their conduction velocities they were grouped as C or Adelta fibers. Bladder filling parameters and afferent nerve spike rate were simultaneously recorded 30 min before administration of saline (nine fibers) or oxybutynin (15 fibers, 1 mg/kg), and again 30, 60, 90, 120, and 150 min after systemic saline or drug administration. RESULTS: No change in C or Adelta afferent spike rate was observed after saline injection (P > 0.90). In the study group, a decrease in afferent activity was noted after systemic administration of oxybutynin for C fibers, which were statistically significant 90 (P < 0.004) and 120 min (P < 0.028) after drug delivery. After 150 min, the spike rate was still lower compared to the baseline filling, without reaching the level of significance (P > 0.09). For the Adelta fibers the decrease in afferent spike rate was already significant at 30 min (P < 0.005) and remained significant during all subsequent fillings (P < 0.012). To avoid a possible confounding influence of the bladder compliance, which increased significantly after injection of oxybutynin (P < 0.011), afferent activity during bladder filling was recalculated. Normalized afferent sensitivity of C and Adelta fibers decreased significantly after injection of oxybutynin. This means that the decrease in afferent spike rate is not the result of an increased compliance. CONCLUSIONS: The findings of this study strongly suggest that oxybutynin directly or indirectly influences bladder sensory nerves, inhibiting the afferent part of the micturition reflex. PMID- 16372317 TI - Does the cystometric filling rate affect the afferent bladder response pattern? A study on single fibre pelvic nerve afferents in the rat urinary bladder. AB - AIMS: To study the effect of physiological and supraphysiological filling rates on the response pattern of single fibre pelvic nerve afferents of the rat urinary bladder. MATERIALS AND METHODS: A total of 37 single afferent bladder units were identified electrophysiologically and their mechanosensitive properties studied. Afferent activity of 13 units was studied at filling rates of 40 and 200 microl/min. Afferent activity of the other units was studied at 200 and 400 microl/min. RESULTS: At the physiological filling rate of 40 microl/min two clearly different types of response pattern were noted. However, at higher supraphysiological filling rates, all units exhibited nearly the same pattern. The difference in mechanosensitive properties of the units that showed a change in response pattern at the supraphysiological filling rate, was characterized by a decrease in pressure at which afferent firing rate peaked. For all units it was found that an increase in filling rate induced an increase in the activation pressure threshold for afferent units, whereas the afferent firing rate at all pressures decreased. CONCLUSIONS: In rats supraphysiological filling rates delay afferent activation, lower afferent firing activity and even change the characteristics of some afferents completely. These data may elucidate some of the differences between ambulatory and conventional urodynamics observed in man. PMID- 16372318 TI - Novel Lamp-2 gene mutation and successful treatment with heart transplantation in a large family with Danon disease. AB - Lysosome-associated membrane protein-2 deficiency (LAMP-2 deficiency), or Danon disease, is a rare X-linked lysosomal disease characterized by cardiomyopathy, vacuolar myopathy, and mental retardation. Less than 20 families with mutations of the Lamp-2 gene have been reported. We describe a family from Sardinia with eight affected patients (4 females and 4 males) and a novel mutation in exon 2 of the Lamp-2 gene (c.102_103delAG). Females developed isolated cardiomyopathy in adulthood, whereas males presented with cardiomyopathy, myopathy, and mental retardation before the age of 20 years. Cardiomyopathy was lethal in three females in their 40s and in three males before the age 20 years. One patient was successfully treated by heart transplantation with more than 5-year follow-up. This study demonstrates that Danon disease is a frequently fatal condition that is potentially treatable with heart transplantation. PMID- 16372319 TI - Effects of temperature on vibration-induced damage in nerves and arteries. AB - Vasospastic episodes in hand-arm vibration syndrome are more prevalent among power-tool workers in cold climates. To test whether cold enhances vibration induced damage in arteries and nerves, tails of Sprague-Dawley rats were vibrated at room temperature (RT) or with tail cooling (<15 degrees C). Cold vibration resulted in a colder tail than either treatment alone. Vibration at both temperatures reduced arterial lumen size. RT vibration generated more vacuoles in arteries than cold vibration. Vibration and cold induced nitration of tyrosine residues in arteries, suggesting free-radical production. Vibration and cold generated similar percentages of myelinated axons with disrupted myelin. Cold with and without vibration caused intraneural edema and dilation of arterioles and venules with blood stasis, whereas vibration alone did not. The similarities, differences, and interactive effects of cold and vibration on nerve and artery damage indicate that temperature is involved mechanistically in the pathophysiology of hand-arm vibration syndrome. PMID- 16372320 TI - Analysis of histopathologic and molecular pathologic findings in Czech LGMD2A patients. AB - Limb-girdle muscular dystrophy type 2A (LGMD2A) is an autosomal-recessive disorder characterized by selective atrophy and progressive weakness of proximal girdle muscles. LGMD2A, the most prevalent form of LGMD, is caused by mutations in the CAPN3 gene that encodes the skeletal muscle-specific member of the calpain family, calpain-3 (p 94). We examined the histopathologic and molecular pathologic findings in 14 Czech LGMD2A patients. Analysis of the CAPN3 gene was performed at the mRNA level, using reverse transcription-polymerase chain reaction (RT-PCR) and sequencing, and/or DNA level, using PCR and denaturing high performance liquid chromatography (DHPLC). Our results confirm that mutation 550 delA is the most frequent CAPN3 defect in Czech LGMD2A patients (9 alleles of 28). Furthermore, we established that, in a patient with the 550 delA/R490W genotype, mRNA carrying frameshift mutation 550 delA was not detected, probably due to its degradation by nonsense-mediated mRNA decay. In muscle biopsies of two LGMD2A patients, a neurogenic pattern simulating a neurogenic lesion was observed. Immunoblot analysis revealed the deficiency of p 94 in all genetically confirmed cases of LGMD2A, and secondary dysferlin deficiency was demonstrated on muscle membranes in 6 patients using immunofluorescence. Thus, we find a combination of DNA and mRNA mutational analysis to be useful in the diagnosis of LGMD2A. Moreover, our study expands the spectrum of calpainopathies to cases that simulate a neurogenic lesion in muscle biopsies, and the knowledge of possible secondary deficiencies of muscular proteins also contributes to a diagnosis of LGMD2A. PMID- 16372321 TI - Age-related morphological changes in skeletal muscle cells of acid alpha glucosidase knockout mice. AB - Glycogen storage disease type II (GSDII), caused by a genetic defect in acid alpha-glucosidase (AGLU), leads to a decline in muscle contractility caused by both muscle wasting and a decrease in muscle quality, i.e., force generated per unit muscle mass. A previous study has shown that loss of muscle mass can only explain one-third of the decrease in contractile performance. Here we report on changes in the intramyocellular structural organization in a mouse knockout model (AGLU(-/-) mice) as a possible cause for the decline in muscle quality. Swollen, glycogen-filled lysosomes and centrally localized cores with cellular debris partially contribute to the decline in muscle quality. Altered localization and deposition of cytoskeletal proteins desmin and titin may reflect adaptive mechanisms at the age of 13 months, but a decline in quality at 20 months of age. The early deposition of lipofuscin in AGLU-deficient myocytes (13 months) is most likely a reflection of enhanced oxidative stress, which may also affect muscle quality. These collective findings, on the one hand, may explain the decrease in tissue quality and, on the other, may represent markers for efficacy of therapeutic interventions to restore muscle function in patients suffering from GSDII. PMID- 16372322 TI - Aerobic exercise and muscle metabolism in patients with mitochondrial myopathy. AB - Exercise therapy improves mitochondrial function in patients with mitochondrial myopathy (MM). We undertook this study to determine the metabolic abnormalities that are improved by exercise therapy. This study identified metabolic pathology using (31)P-magnetic resonance spectroscopy and magnetic resonance imaging (MRI) in a group of patients with MM compared to a control group matched for age, gender, and physical activity. We also observed the effect of exercise therapy for 12 weeks on muscle metabolism and physical function in the MM group. During muscle activity, there was impaired responsiveness of the mitochondria to changes in cytosolic adenosine diphosphate concentration, increased dependence on anaerobic energy pathways, and an adaptive increase in proton efflux in patients with MM. Following exercise therapy, mitochondrial function and muscle mass improved without any change in proton efflux rate. These metabolic findings were accompanied by improvements in functional ability. We conclude that there are significant metabolic differences between patients with MM and a control population, independent of age, gender, and physical activity. Exercise therapy can assist in improving mitochondrial function in MM patients. PMID- 16372323 TI - Acute hypoxia limits endurance but does not affect muscle contractile properties. AB - Acute hypoxia causes skeletal muscle dysfunction in vitro, but little is known about its effect on muscle function in vivo. In 10 healthy male subjects, isometric contractile properties and fatigue resistance of the quadriceps muscle were determined during normoxia and hypoxia using electrically evoked and voluntary contractions. The oxygen saturation (SaO(2); 96.9 +/- 0.7 vs. 79.9 +/- 3.0%; P < 0.001) was reduced during hypoxia. The maximal voluntary contraction (MVC), force-frequency relation, and contraction and relaxation times were unaffected by hypoxia. The endurance time of a sustained 30% MVC was reduced in hypoxia (248 +/- 104 vs. 217 +/- 76 s; P < 0.05), but not that of a sustained 70% MVC. Fatigue induced by electrically evoked intermittent contractions was unaltered. Thus, acute hypoxia has no significant impact on contractile properties of skeletal muscle in vivo but causes reduced endurance during low level sustained voluntary contractions. This indicates that skeletal muscle dysfunction during conditions associated with prolonged hypoxemia, except for limited endurance, is not due to acute effects of hypoxemia. PMID- 16372324 TI - Trigeminal sensory neuropathy related to interferon-alpha treatment. PMID- 16372325 TI - Mitochondrial dysfunction and amyotrophic lateral sclerosis. AB - The causes of motor neuron death in amyotrophic lateral sclerosis (ALS) are still unknown. Several lines of evidence suggest that mitochondrial dysfunction may be involved in the pathogenesis of ALS. Biochemical and morphological mitochondrial abnormalities have been demonstrated in postmortem spinal cords of ALS patients. Furthermore, in transgenic mice expressing mutant Cu,Zn-superoxide dismutase (SOD1), the antioxidant enzyme associated with familial ALS (FALS), mitochondrial abnormalities precede the disease onset, suggesting that mitochondrial dysfunction is causally involved in the pathogenesis of SOD1-FALS. Despite this evidence, it is not yet fully understood how mutant SOD1 damages mitochondria. Recent work has demonstrated that a portion of mutant SOD1 is localized in mitochondria, both in transgenic mice and in FALS patients, where it forms proteinaceous aggregates. These findings have opened new avenues of investigation addressing the hypothesis that mutant SOD1 may directly damage mitochondria. Major future challenges will be to better understand the mechanisms and the consequences of mitochondrial dysfunction in ALS. If mitochondrial dysfunction is convincingly involved in ALS pathogenesis, either as a primary cause or as contributing factor, it is likely to become a novel target for therapeutic intervention. PMID- 16372326 TI - Strength training in old age: adaptation of antagonist muscles at the ankle joint. AB - The purpose of this study was to determine whether strength training could reduce the deficit in plantarflexion (PF) maximal voluntary contraction (MVC) torque observed in previous studies in older subjects relative to young adults. Accordingly, the effects of a 6-month strength training program on the muscle and neural properties of the major muscle groups around the ankle were examined. PF and dorsiflexion (DF) isometric MVC torques were measured and surface electromyographic activity of the triceps surae and tibialis anterior muscles was recorded. The strength training program was very effective in improving strength in PF (+24.5%), and it thus reduced the DF-to-PF MVC torque ratio; in addition, it also induced gains in DF (+7.6%). Thus, there must be an improvement in ankle joint stability. In PF, gains were due particularly to a modification of the agonist neural drive; in DF, the gains appeared to be the consequence of a reduction in antagonist coactivation. Our findings indicate that the investigation of one muscle group should always be accompanied by examination of its antagonist muscle group. PMID- 16372327 TI - Murine androgen-independent neuroendocrine carcinoma promotes metastasis of human prostate cancer cell line LNCaP. AB - BACKGROUND: Although neuroendocrine (NE) cells in prostate cancer have been speculated to accelerate the growth and progression of surrounding cancer cells, the evidence is as yet inconclusive. We investigated the effect of an NE allograft (NE-10) and its cell line, NE-CS, which were established from the prostate of the LPB-Tag 12T-10 transgenic mouse, on human prostate cancer cell line LNCaP. METHODS: The proliferation and pulmonary metastasis of LNCaP xenografts in athymic mice with and without NE-10 allografts were evaluated. Boyden chamber assay and microarray analysis were performed to investigate changes in invasion/migration and mRNA of LNCaP cells under the influence of the NE cells, respectively. RESULTS: NE-10 did not influence the proliferation of LNCaP. The pulmonary metastasis of LNCaP with NE-10 significantly increased compared to mice without it. The NE-CS cells accelerated the in vitro invasion/migration of adenocarcinoma cells. Increased expression of mRNA of gelsolin was observed in LNCaP cells incubated with the supernatant of NE-CS cells. CONCLUSIONS: The NE-10 allograft promotes pulmonary metastasis of subcutaneously inoculated LNCaP cells by facilitating cell invasion. Secretions from NE cells upregulate the expression of gelsolin, which is an actin-binding protein, resulting in acceleration of the migration of LNCaP cells. PMID- 16372328 TI - Prostatic fluid concentrations of isoflavonoids in soy consumers are sufficient to inhibit growth of benign and malignant prostatic epithelial cells in vitro. AB - BACKGROUND: The differential intestinal metabolism of the soy isoflavones is likely to influence the ability of soy to prevent prostate cancer. While daidzein, genistein, and equol have direct antiproliferative effects on prostatic epithelial cells in vitro, there are no such data for the isoflavone glycitein, or seven metabolites: O-desmethylangolensin (ODMA), 6-hydroxyODMA (6H-ODMA), dihydrodaidzein (DHD), cis-4-hydroxyequol (C4HE), 3'-hydroxydaidzein (3HD), 6 hydroxydaidzein (6HD), and 8-hydroxydaidzein (8HD). In the current study, the in vitro activities of these compounds were elucidated, and the active ranges of concentrations were compared to that found in Caucasian prostatic fluid (PF) and plasma samples. METHODS: The effects of isoflavonoids on cell growth, cell cycle distribution, and apoptosis (active Caspase 3) were examined on benign prostatic epithelial cells (PrEC), and the prostate cancer cell line LNCaP. RESULTS: PF concentrations of genistein, equol, and daidzein (but not ODMA or DHD) were often within the ranges that reduce PrEC growth in vitro. Profound differences in sensitivities were observed with LNCaP. The hydroxydaidzeins, C4HE, and 6H-ODMA had significant inhibitory effects at 10(-5)M on PrEC growth (but not LNCaP). Glycitein had significant effects on both. Reductions in cell growth were typically associated with both changes in cell cycle distribution and Caspase 3 activation. When five isoflavonoids were used in combination at concentrations present in PF samples, synergistic effects were observed. CONCLUSION: The profound differences in sensitivities of prostatic epithelial cells to these compounds along with their synergistic effects suggest that multiple metabolites in vivo may be optimal for preventing prostate cancer. PMID- 16372329 TI - Superoxide dismutase in the prostate lobes of aging Brown Norway rats. AB - BACKGROUND: Spontaneous age-dependent epithelial cell hyperplasia occurs in the lateral and dorsal, but not the ventral, lobes of aging Brown Norway (BN) rats. Diminished antioxidant enzyme activities and increased formation of reactive oxygen species (ROS) promote the pathology of many aging disorders. We investigated the hypothesis that prostatic epithelial cell hyperplasia in the BN rat was related to age-dependent and/or lobe-specific changes in superoxide dismutase (SOD). MATERIALS AND METHODS: Using Western blots, immunohistochemistry and enzyme activity assays we determined the levels of protein expression, subcellular localization, and activities, respectively, of the three SOD isoforms, cytoplasmic SOD1, mitochondrial SOD2, and extracellular SOD3 in the ventral, lateral, and dorsal prostate lobes of 4-month-old rats with normal prostate morphology, in 24-month-old rats with lobe-specific hyperplasia and in older 30-month-old rats. RESULTS: We observed little change in SOD activities as a function of age, although expression of SOD3 increased in the prostatic lobes of older rats. SOD2 levels were higher in the lateral lobe of 4- and 24-month-old rats, but declined by 30 months of age to levels in the ventral and dorsal lobes. SOD1 was localized by immunohistochemistry to the nuclei of epithelial cells in all lobes, but the number of immunopositive nuclei increased in the lateral and dorsal lobes of 24-month-old animals. The concentration of zinc was highest in the prostate lobes of 24-month-old animals. CONCLUSION: Based upon our data, superoxide dismutase is not significantly altered in the rat prostate during aging and thus is unlikely to be an important factor in the evolution of epithelial cell hyperplasia. PMID- 16372330 TI - Enhancement of intermittent androgen ablation by "off-cycle" maintenance with finasteride in LNCaP prostate cancer xenograft model. AB - BACKGROUND: Intermittent androgen ablation (IAA) was developed with the intention of delaying progression of prostate cancer to androgen-independence and improving quality of life. Our previous studies suggest that relative to dihydrotestosterone (DHT), testosterone (T) is a weak inducer of proliferation and a more potent inducer of differentiation. We hypothesize that administration of finasteride (F), a type-II 5-alpha-reductase inhibitor that increases T and decreases DHT, during the IAA "off-cycle" would enhance the efficacy. METHODS: After LNCaP tumor establishment, nude mice were castrated and randomized to continuous androgen ablation (CAA), continuous androgen ablation plus finasteride (CAA + F), intermittent androgen ablation (IAA), or intermittent androgen ablation plus finasteride (IAA + F). RESULTS: After one cycle of therapy, mice treated with IAA + F had significantly less tumor growth than the other treatment groups (P = 0.002). Mice treated with IAA + F had the best survival (P = 0.048) and were 3-5 times more likely to be alive 70 days following treatment initiation. CONCLUSIONS: IAA with finasteride provides the most favorable tumor growth kinetics and survival compared to both CAA and standard IAA. PMID- 16372331 TI - Prognostic value of PINP, bone alkaline phosphatase, CTX-I, and YKL-40 in patients with metastatic prostate carcinoma. AB - BACKGROUND: To examine the prognostic value of markers of bone metabolism (serum PINP, BAP, and CTX-I) and serum YKL-40 in metastatic prostate carcinoma (PC). METHODS: The biomarkers were determined by ELISAs in 153 metastatic PC patients before treatment with parenteral estrogen or total androgen ablation. The median follow-up was 4.9 years. One hundred fifteen patients died. RESULTS: The biomarkers were increased in the patients compared to controls (P < 0.001), and related to performance status and Soloway score (except YKL-40), but not to T category and WHO tumor grade. PINP was elevated in 87%, BAP (55%), CTX-I (33%), and YKL-40 (43%). Univariate analysis showed an association to survival: PINP (HR = 1.6, P < 0.0001), BAP (HR = 1.4, P < 0.0001), CTX-I (HR = 1.7, P < 0.0001), and YKL-40 (HR = 1.4, P = 0.004). In multivariate Cox analysis performance status, WHO grade, Soloway score, PINP, and YKL-40 were independently predictive factors. CONCLUSIONS: High serum PINP, BAP, CTX-I, and YKL-40 are associated with poor outcome of metastatic PC patients. PMID- 16372332 TI - Piloting the zebrafish genome browser. AB - This correspondence is a primer for the zebrafish research community on zebrafish tracks available in the UCSC Genome Browser at http://genome.ucsc.edu based on Sanger's Zv4 assembly. A primary capability of this facility is comparative informatics between humans (as well as many other model organisms) and zebrafish. The zebrafish genome sequencing project has played important roles in mutant mapping and cloning, and comparative genomic research projects. This easy-to-use genome browser aims to display and download useful genome sequence information for zebrafish mutant mapping and cloning projects. Its user-friendly interface expedites annotation of the zebrafish genome sequence. PMID- 16372333 TI - PAC1-R null isoform expression in human prostate cancer tissue. AB - BACKGROUND: PACAP is a member of the VIP/GHRH family of neuropeptides and has important effects on prostate cell proliferation. Here we analyze the expression and localization of PACAP and its specific receptor variants (PAC(1)-R) in tissues collected from patients undergoing prostate biopsy and surgery for benign prostatic hyperplasia (BPH) and prostate cancer (PCa). METHODS: Reverse transcriptase (RT)-polymerase chain reaction (PCR), DNA sequencing, and immunohistochemistry. RESULTS: PACAP and PAC(1)-R were localized by immunohistochemistry in the prostate tissue. While in healthy and BPH tissues PAC(1)-R positive staining is present in all the epithelial cells lining the lumen of the acini and in some stromal cells (mostly in the apical portion of the cells), in PCa tissues, anti-PAC(1)-R antibody stained the apical portion of the cells. We provide evidence that PAC(1)-R null and SV(1)/SV(2) variants are all present in normal and hyperplastic tissues, while in PCa tissue PAC(1)-R null is the most relevant receptor variant expressed. CONCLUSIONS: Our data demonstrates that the PAC(1)-R null variant is the most relevant isoform expressed in human PCa tissue being suggestively related with the events determining the outcome of prostate cancer. PMID- 16372334 TI - E-cadherin polymorphisms and haplotypes influence risk for prostate cancer. AB - BACKGROUND: The E-cadherin (CDH1) gene has been implicated in prostate cancer (PCA) risk, however, the exact mechanism is unknown. Several polymorphisms, such as the C/A variant -160 base pairs from the transcription start site, in the CDH1 gene promoter region have been associated with cancer risk, mainly in European descent populations. METHODS: We screened the entire coding region and 3.0 kilobases of the CDH1 promoter for polymorphisms in 48 African Americans using dHPLC. Twenty-one (21) polymorphisms were observed. Four polymorphisms, including -160C/A, were genotyped in a genetic association study using incident PCA cases (N = 427) and unaffected controls (N = 337) of similar age from three different ethnic groups consisting of African Americans, Jamaicans, and European Americans. RESULTS: We observed a significantly higher frequency of the -160A allele among European American PCA patients (27.5%) compared to the control group (19.7%) (P = 0.04). More importantly, among men of European ancestry under the age of 65 who possess the -160 A allele there was over three times increased risk for prostate cancer (P = 0.05). Also, the AACT haplotype bearing the -160A allele was significantly associated with PCA in European Americans (P = 0.04). CONCLUSIONS: Our data indicate that CDH1 likely is a low-penetrant PCA susceptibility gene, however, population differences in linkage disequilibrium within the CDH1 gene region may influence the effect of susceptibility alleles such as -160A. PMID- 16372335 TI - Characterization of an antibody that can detect the Kai1/CD82 murine metastasis suppressor. AB - BACKGROUND: Kai1, also known as CD82, is a member of the tetraspanin family (TM4SF). The human homolog, KAI1, is an activation antigen of T-cells and is a metastasis suppressor for prostate and other cancers. Little is known about the mouse protein because of the lack of antibody reagents. METHODS: Peptide immunized rabbits were used to generate polyclonal antibody to Kai1. The antibody was analyzed using immunoblotting, flow cytometry, and immunohistochemistry. RESULTS: This antibody specifically recognizes murine Kai1 protein, crossreacts with rat Kai1 but not with human KAI1. The normal tissue distribution of this protein in mice is shown to be similar to that of the human homolog. Interestingly, mouse prostatic epithelium showed differential expression within the lobes. CONCLUSION: This antibody, the first described that can specifically detect murine Kai1/CD82, should be very useful in addressing the mechanism of action of Kai1 in metastatic suppression. PMID- 16372336 TI - Start me up: cell signaling and the journey from oocyte to embryo in C. elegans. AB - Intercellular communication plays a pivotal role in regulating and coordinating oocyte meiosis and fertilization, key triggers for embryonic development. The nematode Caenorhabaditis elegans has emerged as an important experimental paradigm for exploring these fundamental reproductive processes and their regulation. The oocytes of most animal species arrest during meiotic prophase and complete meiosis in response to intercellular signaling in the process of meiotic maturation. Oocyte meiotic maturation is defined by the transition between diakinesis and metaphase of meiosis I and is accompanied by nuclear envelope breakdown and meiotic spindle assembly. As such, the meiotic maturation process is essential for completing meiosis and a prerequisite for successful fertilization. In C. elegans, the processes of meiotic maturation, ovulation, and fertilization are temporally coupled: sperm utilize the major sperm protein as a hormone to trigger oocyte meiotic maturation, and, in turn, the maturing oocyte signals its own ovulation, leading to fertilization. The powerful genetic screens possible in C. elegans have led to the identification of several sperm cell surface proteins that are required for the interaction and fusion of gametes at fertilization. The study of these proteins provides fundamental insights into fertilization mechanisms, their role in speciation, and their potential conservation across phyla. Signaling processes sparked by fertilization are required for meiotic chromosome segregation and initiating the embryonic program. Here we review recent advances in understanding how signaling mechanisms contribute to the oocyte-to-embryo transition in C. elegans. PMID- 16372337 TI - In vitro culture of embryos of the guppy, Poecilia reticulata. AB - The rich variation in adult color patterns of male guppies (Poecilia reticulata) has attracted the attention of geneticists and ecologists for almost a century. Studies on their embryogenesis, however, have been limited by the fact that guppies are live bearers. We have observed normal development after explantation of guppy embryos from the ovary of pregnant females at various times after last parturition, and found that development of each batch of eggs is slightly asynchronous, most likely due to asynchronous fertilization. We have cultured explanted embryos in vitro and continuously observed their development. Although embryos explanted a few days after fertilization survived up to 4 weeks in culture, they did not complete their development. In contrast, embryos explanted at late stages of gestation could hatch and develop to fertile adults. Our embryo culture techniques overcome some of the limitations of using livebearers as study objects, and they allow continuous observation of and accessibility to live embryos at all stages. PMID- 16372338 TI - A new device for cricothyroidotomy. AB - A new instrument for coniotomy was developed and tested with cadaver experiments. The instrument should be simple, easy to handle, and should not have the various drawbacks of the existing devices. The new instrument was tested by 17 medical students and medical doctors on fresh human cadavers. We measured the time necessary for performing a successful experiment. It lasted 5-28 sec (mean value 22 sec) depending on the experience of the people carrying out the tests. This time is much shorter than with other existing instruments tested under experimental conditions. The instrument consists of a tube and a trocar with a small knife on the tip. The knife is used to incise the skin over the cricothyroid ligament and to perforate this ligament in sagittal position. The tube is provided with a plastic stripe, which limits the depth of the entrance of the instrument into the larynx and can also be used to attach the tube to the neck of the patient. After entering the larynx, the tube can be connected to an inflating bag. The correct seat of the tube was tested by inflation, with a calibrated probe or with laryngoscopy. The results of our tests encourage the development of this new instrument for industrial production. PMID- 16372339 TI - Seven renal arteries. PMID- 16372340 TI - Tissue inhibitor of metalloproteinase 1 regulates matrix metalloproteinase activity during newt limb regeneration. AB - Matrix metalloproteinase (MMP) activity is important for newt limb regeneration. In most biological processes that require MMP function, MMP activity is tightly controlled by a variety of mechanisms, including the coexpression of natural inhibitors. Here, we show that gene expression of one such inhibitor, tissue inhibitor of metalloproteinase 1 (NvTIMP1), is upregulated during the wound healing and dedifferentiation stages of regeneration when several MMPs are at their maximal expression levels. Newt MMPs and NvTIMP1 also exhibit similar spatial expression patterns during the early stages of limb regeneration. NvTIMP1 inhibits the proteolytic activity of regeneration-related newt MMPs and, like human TIMP1, can induce a weak mitogenic response in certain cell types. These results suggest that NvTIMP1 may be functioning primarily to maintain optimal levels of MMP activity during the early stages of limb regeneration, while possibly serving a secondary role as a mitogen. PMID- 16372341 TI - Quantitative evaluation of the accuracy of micro-computed tomography in tooth measurement. AB - Micro-computed tomography (MCT) is a noninvasive technique for visualizing morphological characteristics of teeth in a detailed and accurate manner, without causing any tooth destruction. A large amount of information can be obtained from such scans. The slices can be recreated in any plane, and the data can be represented as either two-dimensional (2D) or three-dimensional (3D) images. In addition, the internal and external anatomy can be demonstrated simultaneously or separately, and the images can be assessed qualitatively and quantitatively. However, only the qualitative accuracy of MCT had been evaluated previously. The aim of the present study was to evaluate the accuracy of MCT quantitatively by comparing the values obtained by MCT with those of other imaging methods. Twelve teeth were scanned using a desktop X-ray micro-CT scanner, and the images were reconstructed and measured. Values obtained by direct measurement served as the primary reference for linear measurements. Measurements made by a 3D scanner and by photography were additional references. MCT was found to be a reliable method of making linear measurements and may be a useful device for measuring distance and for observing both internal and external tooth structure using the reconstructed 3D form. PMID- 16372342 TI - A novel interactive anatomic atlas of the hand. AB - Classical anatomic atlases cannot provide the spectrum of views and the detail required in modern diagnostic and surgical techniques. Computer modeling opens the possibility to choose any view from one single model. A computerized model of the hand is presented, which has been obtained by segmentation and graphic modeling of the Visible Human dataset. In addition to being able to choose arbitrary viewpoints, it allows interrogation of the chosen views by mouse click. We believe the functions of these new kinds of atlases are superior to the classical ones. PMID- 16372343 TI - Persistent left superior vena cava. AB - Congenital venous anomalies are relatively common and some have clinical implications. An example of persistent left superior vena cava was found during a routine dissection. This vein was carefully dissected and followed to its termination in the right atrium. The same cadaver also presented four branches from the arch of the aorta. A left superior vena cava occurs in early development but disappears later. The clinical significance of such a persistent left superior vena cava assumes importance during cardiac catheterization via the left subclavian vein. PMID- 16372344 TI - Surgically important variations of the jugular veins. AB - Knowledge of variations of veins of head and neck in relation to external jugular, anterior jugular, internal jugular, and facial veins is important to surgeons doing head and neck surgery as well as to radiologists doing catheterization and to clinicians in general. In the current case, multiple variations in the veins of the left side of neck are reported. The anterior division of retromandibular vein was absent. The facial vein continued as anterior jugular vein. The internal jugular vein was duplicated above the level of hyoid bone. There was a large communicating vein between the anterior jugular vein and anterior division of internal jugular vein. Lingual vein drained into the communicating vein. Jugular venous arch was abnormally large, doubled, and highly placed. The veins of the right side were normal. PMID- 16372345 TI - The axillary sheath and single-injection axillary block. AB - Failure to block the terminal nerves of the brachial plexus, the circumferential type of incomplete axillary brachial plexus block, is the main problem of the single-injection technique. Two studies were carried out to observe the internal anatomy of the axillary sheath and the effect of different volumes of dye injected into the sheath in cadavers. In our first study, the internal arrangement of the axillary sheath and its septae were examined microscopically by slicing the sheath longitudinally and transversely. In the second study, boluses of 10, 20, and 40 cc of methylene-blue were injected into one of the compartments of the axillary sheath. The axillary sheath was dissected out and sliced transversely to observe the spread of the dye in the injected and in the adjacent compartments. The specimens of the axillary sheath were then opened longitudinally and the septae excised and examined at x10 magnification to see the effect of the various volumes of the injection bolus. This study shows that septae from the deep surface of the axillary sheath form compartments for each nerve. The septae function as barriers under physiologic conditions. By increasing the injected volume of solutions, bubble-like defects are produced in the septae in the compartments into which leakage was demonstrated. PMID- 16372346 TI - Mass-dependent decline of skeletal muscle function in cancer cachexia. AB - CD2F1 mice were inoculated with C26 adenocarcinoma cells, followed by assessment of ex vivo muscular function. Muscles from tumor-bearing mice had a significantly lower force output during a single maximal contraction and during repeated contractions than control muscles. The relative force output, however, did not differ when corrected for muscle mass. Thus, cachexia significantly reduces absolute skeletal muscle function, but muscle "quality" appears unaltered. PMID- 16372347 TI - A novel MSH2 germline mutation in homozygous state in two brothers with colorectal cancers diagnosed at the age of 11 and 12 years. AB - Hereditary non-polyposis colorectal cancer (HNPCC) syndrome is caused by heterozygous germline mutations in DNA mismatch repair genes (MMR), (MSH2, MLH1, MSH6, and PMS2) and it is inherited in an autosomal dominant pattern with high penetrance. Several patients have been reported carrying bi-allelic MMR gene mutations and whose phenotype resembled a syndrome with childhood malignancies including hematological malignancies, brain, and colorectal tumors. This phenotype is similar to the tumor spectrum of MMR knockout mice. Herein we describe two brothers of healthy consanguineous parents from Pakistan, who had developed two and three colorectal cancers at the ages of 11 and 12 years, respectively, and less than 30 polyps. Tumor specimens were microsatellite instable (MSI-H), and expression of MSH2 and MSH6 was lost. Mutation analyses of DNA samples from both patients revealed a novel homozygous c.2006-5T > A mutation in intron 12 of the MSH2 gene. This phenotype of the brothers is unusual as they neither develop hematological malignancies nor brain tumors at an older age of presentation than other patients with homozygous MSH2 mutations. The milder phenotype may be due to the expression of low amounts of MSH2 protein with reduced activity. PMID- 16372348 TI - Dpp signaling down-regulates the expression of VM32E eggshell gene during Drosophila oogenesis. AB - Among the members of the Drosophila melanogaster vitelline membrane protein gene family, VM32E has the unique feature of being a component of both the vitelline and the endochorion layers. The VM32E gene is expressed at stage 10 of egg chamber development in the main body follicle cells, and it is repressed in the anterior and posterior follicle cells. Here, we show that this spatial restriction of VM32E gene expression is conserved in the D. pseudoobscura orthologous gene, suggestive of a conserved function of VM32E protein. The VM32E gene is not expressed in the centripetal migrating follicle cells, where the Decapentaplegic (Dpp) pathway is active in patterning the anterior eggshell structures. By analyzing the native VM32E gene and the activity of specific VM32E regulatory regions, in genetic backgrounds altering the Dpp pathway, we show that VM32E gene is negatively regulated by the Dpp signaling. Therefore, it appears that the Dpp signaling pathway executes its control on eggshell morphogenesis also by controlling the expression of eggshell structural genes. PMID- 16372349 TI - Accounting for loop flexibility during protein-protein docking. AB - Although reliable docking can now be achieved for systems that do not undergo important induced conformational change upon association, the presence of flexible surface loops, which must adapt to the steric and electrostatic properties of a partner, generally presents a major obstacle. We report here the first docking method that allows large loop movements during a systematic exploration of the possible arrangements of the two partners in terms of position and rotation. Our strategy consists in taking into account an ensemble of possible loop conformations by a multi-copy representation within a reduced protein model. The docking process starts from regularly distributed positions and orientations of the ligand around the whole receptor. Each starting configuration is submitted to energy minimization during which the best-fitting loop conformation is selected based on the mean-field theory. Trials were carried out on proteins with significant differences in the main-chain conformation of the binding loop between isolated form and complexed form, which were docked to their partner considered in their bound form. The method is able to predict complexes very close to the crystal complex both in terms of relative position of the two partners and of the geometry of the flexible loop. We also show that introducing loop flexibility on the isolated protein form during systematic docking largely improves the predictions of relative position of the partners in comparison with rigid-body docking. PMID- 16372350 TI - A dynamic N-capping motif in cytochrome b5: evidence for a pH-controlled conformational switch. AB - Apocytochrome b5 is a marginally stable protein exhibiting under native conditions a slow conformational exchange in its C-terminal region. The affected elements of secondary structure include a 3(10)-helix containing at its N terminus a histidine Ncap and a subsequent proline. Participation of the neutral histidine side-chain in backbone amide capping lowers the imidazole pKa. To explore the nature of the conformational exchange in the protein and determine whether it is related to cis-trans isomerization of the His-Pro bond, three octapeptides encompassing the helix were synthesized and studied by NMR spectroscopy. One corresponded to the wild-type sequence, the second was the D histidine epimer, and the third contained an alanine in place of the proline. It was found that the rates of cis-trans interconversion in the proline-containing peptides were slower than the rates of the conformational exchange in the protein. In addition, the wild-type peptide hinted at a predisposition for Ncap formation when in the trans configuration. Analysis of the pH response of the peptides and protein suggested that at pH near neutral, the conformational exchange detected in the protein involved only species with a trans His-Pro bond and could be approximated with a three-state model by which the terminal helix sampled a locally unfolded state. This state, which contained an uncapped histidine with a normal pKa, partitioned into neutral and protonated populations according to pH. The intrinsic conformational bias of the wild-type peptide and the pH-driven equilibria illustrated how a 3(10)-element could serve as a nucleation site for structural rearrangement. PMID- 16372351 TI - HRAS mutations in Costello syndrome: detection of constitutional activating mutations in codon 12 and 13 and loss of wild-type allele in malignancy. AB - Costello syndrome (CS) is a complex developmental disorder involving characteristic craniofacial features, failure to thrive, developmental delay, cardiac and skeletal anomalies, and a predisposition to develop neoplasia. Based on similarities with other cancer syndromes, we previously hypothesized that CS is likely due to activation of signal transduction through the Ras/MAPK pathway [Tartaglia et al., 2003]. In this study, the HRAS coding region was sequenced for mutations in a large, well-characterized cohort of 36 CS patients. Heterogeneous missense point mutations predicting an amino acid substitution were identified in 33/36 (92%) patients. The majority (91%) had a 34G --> A transition in codon 12. Less frequent mutations included 35G --> C (codon 12) and 37G --> T (codon 13). Parental samples did not have an HRAS mutation supporting the hypothesis of de novo heterogeneous mutations. There is phenotypic variability among patients with a 34G --> A transition. The most consistent features included characteristic facies and skin, failure to thrive, developmental delay, musculoskeletal abnormalities, visual impairment, cardiac abnormalities, and generalized hyperpigmentation. The two patients with 35G --> C had cardiac arrhythmias whereas one patient with a 37G --> T transversion had an enlarged aortic root. Of the patients with a clinical diagnosis of CS, neoplasia was the most consistent phenotypic feature for predicating an HRAS mutation. To gain an understanding of the relationship between constitutional HRAS mutations and malignancy, HRAS was sequenced in an advanced biphasic rhabdomyosarcoma/fibrosarcoma from an individual with a 34G --> A mutation. Loss of the wild-type HRAS allele was observed, suggesting tumorigenesis in CS patients is accompanied by additional somatic changes affecting HRAS. Finally, due to phenotypic overlap between CS and cardio-facio-cutaneous (CFC) syndromes, the HRAS coding region was sequenced in a well-characterized CFC cohort. No mutations were found which support a distinct genetic etiology between CS and CFC syndromes. PMID- 16372353 TI - Cough-induced hemiplegic migraine with impaired consciousness in cystic fibrosis. AB - The coughing paroxysms of patients with cystic fibrosis may occasion neurological symptoms. Although cough syncope is well-known, and is associated with headache and paralysis, a migrainous mechanism has not been reported. We reviewed the medical records, autonomic testing results, and responses to treatment in two cystic fibrosis patients with similar presentations of cough-induced impairment of consciousness followed by headache and paralysis. A 24-year-old woman and an unrelated 38-year-old man, both with cystic fibrosis, developed post-tussive neurologic deficits. Both patients reported infrequent dramatic spells, always preceded by major hemoptysis, and associated with left-sided paralysis, transient blindness, nausea, and severe pulsating headaches. Autonomic testing demonstrated only postural tachycardia and a near-vasodepressor episode in the woman, and mild, generalized sympathetic dysfunction in the man. Treatment for presumptive migraine with aura with verapamil nearly eradicated symptoms in both patients. Discontinuation of verapamil in the woman was associated with symptom recurrence and a stroke, with significant persistent residual left hemiparesis. In conclusion, cough-induced neurologic deficits were previously reported with cystic fibrosis, without clear understanding of the mechanism of impairment of consciousness. Based on the hemiparesis, nausea, and throbbing headache, which repeatedly followed the events in both patients, and based on the response to verapamil, we hypothesize a migrainous mechanism in both of our patients. The pathophysiology that links the hemoptysis to the spells deserves further investigation. PMID- 16372352 TI - Safety and efficacy of recombinant alpha(1)-antitrypsin therapy in cystic fibrosis. AB - Neutrophil elastase (NE) is thought to be the most important protease which damages the cystic fibrosis (CF) lung. Attempts have been made to suppress this activity using the plasma-derived inhibitor, alpha(1)-antitrypsin (AAT). In this pilot study, the safety and efficacy of inhaled recombinant human AAT (rAAT) as a treatment for CF were investigated. Thirty-nine patients participated in a prospective, double-blinded, randomized, placebo-controlled phase II trial to examine the effect of rAAT (500, 250, and 125 mg) on sputum NE activity. Sputum myeloperoxidase (MPO), interleukin-8, tumor necrosis factor receptors, sputum and plasma NE/AAT complexes, and safety parameters were also measured. Subjects were randomized to receive nebulized treatment once a day for 4 weeks, followed by 2-4 weeks with no study treatment, and then a 2-week rechallenge phase. Trends toward a reduction in NE activity were observed in patients treated with 500 mg and 250 mg of rAAT compared to placebo. Sputum NE/AAT complex and MPO levels were lower on rAAT compared to placebo. No major adverse events and, in particular, no allergic reactions to rAAT were observed. Although significant differences between rAAT and placebo for sputum NE activity were not observed, some improvements were found for secondary efficacy variables. This study demonstrated that nebulized rAAT is safe and well-tolerated, but has a limited effect on NE activity and other markers of inflammation. PMID- 16372354 TI - Measuring exhaled breath condensates in infants. AB - There is growing interest in investigating compounds of exhaled breath condensates (EBC) as potential noninvasive markers of airways disease processes. Some of these markers have the potential to provide information on the early stages of disease. In this paper, we present a method for collecting EBC during both oral and nasal breathing in infants. Fifty-four infants (mean age, 13.3 months; range, 1-30 months) undergoing infant lung-function testing were recruited for this study. Breath condensates were collected during sedated sleep, using a custom-made collection device. Collections were made for 10 min during normal tidal breathing. Nasal measurements were attempted in all children by placing a face-mask over the nose and mouth and keeping the mouth closed. In 14 infants, oral measurements were made by placing a face-mask over the mouth only and occluding the nose. Condensates were collected successfully in all but one child. The collected volume ranged from 50-550 microl (mean +/- SD, 281.8 +/- 145.8 microl). The volume of EBC collected was correlated to age, length, weight, and minute ventilation. Significantly more EBC was collected during oral compared to nasal breathing (354.3 vs. 277.5 microl, P=0.03). There were no significant changes in heart rate, respiratory rate, or oxygen saturation during collection. The collection of EBC in young children and infants is feasible and safe, and the method used here allows the successful collection of reasonable amounts of exhaled condensate. PMID- 16372355 TI - Edge-count probabilities for the identification of local protein communities and their organization. AB - We present a computational approach based on a local search strategy that discovers sets of proteins that preferentially interact with each other. Such sets are referred to as protein communities and are likely to represent functional modules. Preferential interaction between module members is quantified via an analytical framework based on a network null model known as the random graph with given expected degrees. Based on this framework, the concept of local protein community is generalized to that of community of communities. Protein communities and higher-level structures are extracted from two yeast protein interaction data sets and a network of published interactions between human proteins. The high level structures obtained with the human network correspond to broad biological concepts such as signal transduction, regulation of gene expression, and intercellular communication. Many of the obtained human communities are enriched, in a statistically significant way, for proteins having no clear orthologs in lower organisms. This indicates that the extracted modules are quite coherent in terms of function. PMID- 16372356 TI - Prediction of protein stability changes for single-site mutations using support vector machines. AB - Accurate prediction of protein stability changes resulting from single amino acid mutations is important for understanding protein structures and designing new proteins. We use support vector machines to predict protein stability changes for single amino acid mutations leveraging both sequence and structural information. We evaluate our approach using cross-validation methods on a large dataset of single amino acid mutations. When only the sign of the stability changes is considered, the predictive method achieves 84% accuracy-a significant improvement over previously published results. Moreover, the experimental results show that the prediction accuracy obtained using sequence alone is close to the accuracy obtained using tertiary structure information. Because our method can accurately predict protein stability changes using primary sequence information only, it is applicable to many situations where the tertiary structure is unknown, overcoming a major limitation of previous methods which require tertiary information. The web server for predictions of protein stability changes upon mutations (MUpro), software, and datasets are available at http://www.igb.uci.edu/servers/servers.html. PMID- 16372358 TI - Pressure perturbation calorimetry of helical peptides. AB - Pressure perturbation calorimetry quantifies the temperature dependence of a solute's thermal expansion coefficient, providing information about solute solvent interactions. We tested the idea that pressure perturbation calorimetry can provide information about solvent-accessible surface area by studying peptides with different secondary structures. The peptides comprised two host guest series: one predominately an alpha-helix, the other predominately a polyproline II helix. In aqueous buffer, we find a correlation between the amount of secondary structure as assessed by circular dichroism spectropolarimetry and the pressure perturbation calorimetry data. We conclude that pressure perturbation calorimetry can provide information about the exposure of polar and nonpolar surface area. Data acquired in a buffered urea solution, however, are not as easily interpreted. PMID- 16372359 TI - Experimental evaluation of starch utilization mechanism by activated sludge. AB - The study aimed to explore the conversion processes of hydrolysable substrates by activated sludge. Experimental data were collected from a sequencing batch reactor (SBR) and from batch tests using activated sludge acclimated to native potato starch (NPS). Parallel batch tests were run with NPS (particulate), soluble starch (SolS), maltose, and glucose for comparative evaluation. The fate of organic carbon in the reactor was followed directly by measuring substrate, poly-glucose, and oxygen uptake rate. Results indicated that adsorption was the dominant mechanism for starch removal with subsequent enzymatic hydrolysis inside the flocs. The role of bulk liquid enzyme activity was minimal. Starch was observed to hydrolyze to maltose rather than glucose. The behavior of NPS and SolS was quite similar to maltose in terms of poly-glucose formation and oxygen uptake. Since the simplest hydrolysis product was maltose, the biomass was not acclimated to glucose and thus, glucose exhibited a significantly different removal and storage pattern. The study also showed that differentiation of readily biodegradable and slowly biodegradable COD should better be based on the kinetics of their utilization rather than simple physical characterization. PMID- 16372357 TI - Multipass membrane protein structure prediction using Rosetta. AB - We describe the adaptation of the Rosetta de novo structure prediction method for prediction of helical transmembrane protein structures. The membrane environment is modeled by embedding the protein chain into a model membrane represented by parallel planes defining hydrophobic, interface, and polar membrane layers for each energy evaluation. The optimal embedding is determined by maximizing the exposure of surface hydrophobic residues within the membrane and minimizing hydrophobic exposure outside of the membrane. Protein conformations are built up using the Rosetta fragment assembly method and evaluated using a new membrane specific version of the Rosetta low-resolution energy function in which residue residue and residue-environment interactions are functions of the membrane layer in addition to amino acid identity, distance, and density. We find that lower energy and more native-like structures are achieved by sequential addition of helices to a growing chain, which may mimic some aspects of helical protein biogenesis after translocation, rather than folding the whole chain simultaneously as in the Rosetta soluble protein prediction method. In tests on 12 membrane proteins for which the structure is known, between 51 and 145 residues were predicted with root-mean-square deviation <4 A from the native structure. PMID- 16372360 TI - Determination of diffusion coefficients of proteins in stationary phases by frontal chromatography. AB - A strategy to determine effective diffusion coefficients of proteins in chromatographic gels is presented in this article. An experimental methodology based on frontal liquid chromatography was combined with a numerical methodology based on a mathematical model describing the chromatographic process including the extra-column dispersion, the dispersion due to the packed bed, the external mass transfer from the bulk phase to the stationary phase, and the diffusive transport within the stationary phase. The methodology has several advantages compared to previously reported methods to determine diffusion coefficients in that no other equipment than an HPLC is required, any class of stationary phases can be investigated as long as the experiments are performed under non-binding conditions, and no modification, e.g., moulding of slabs or membranes, to the stationary phase is required. To show the applicability of the methodology, the effective diffusion coefficients of lysozyme, bovine serum albumin, and immunoglobulin gamma in Sepharose CL-4B were determined and shown to be comparable with those determined with other methods. PMID- 16372361 TI - The expression of a Pichia stipitis xylose reductase mutant with higher K(M) for NADPH increases ethanol production from xylose in recombinant Saccharomyces cerevisiae. AB - Xylose fermentation by Saccharomyces cerevisiae requires the introduction of a xylose pathway, either similar to that found in the natural xylose-utilizing yeasts Pichia stipitis and Candida shehatae or similar to the bacterial pathway. The use of NAD(P)H-dependent XR and NAD(+)-dependent XDH from P. stipitis creates a cofactor imbalance resulting in xylitol formation. The effect of replacing the native P. stipitis XR with a mutated XR with increased K(M) for NADPH was investigated for xylose fermentation to ethanol by recombinant S. cerevisiae strains. Enhanced ethanol yields accompanied by decreased xylitol yields were obtained in strains carrying the mutated XR. Flux analysis showed that strains harboring the mutated XR utilized a larger fraction of NADH for xylose reduction. The overproduction of the mutated XR resulted in an ethanol yield of 0.40 g per gram of sugar and a xylose consumption rate of 0.16 g per gram of biomass per hour in chemostat culture (0.06/h) with 10 g/L glucose and 10 g/L xylose as carbon source. PMID- 16372362 TI - Pi topology and spin alignment in unique photoexcited triplet and quintet states arising from four unpaired electrons of an organic spin system. AB - Syntheses, electronic structures in the ground state, unique photoexcited states, and spin alignment are reported for novel biradical 1, which was designed as an ideal model compound to investigate photoinduced spin alignment in the excited state. Electron spin resonance (ESR), time-resolved ESR (TRESR), and laser excitation pulsed ESR experiments were carried out. The magnetic properties were examined with a SQUID magnetometer. In the electronic ground state, two radical moieties interact very weakly (almost no interaction) with each other through the closed-shell diphenylanthracene spin coupler. On photoirradiation, a novel lowest photoexcited state with the intermediate spin (S = 1) arising from four unpaired electrons with low-lying quintet (S = 2) photoexcited state was detected. The unique triplet state has an interesting electronic structure, the D value of which is reduced by antiferromagnetic spin alignment between two radical spins through the excited triplet spin coupler. The general theoretical predictions of the spin alignment and the reduction of the fine-structure splitting of the triplet bis(radical) systems are presented. The fine-structure splitting of the unique photoexcited triplet state of 1, as well as the existence of the low-lying quintet state, is interpreted well on the basis of theoretical predictions. Details of the spin alignment in the photoexcited states are discussed. PMID- 16372363 TI - Antianxiety effects of riparin I from Aniba riparia (Nees) Mez (Lauraceae) in mice. AB - This work presents the behavioral effects of riparin I (methyl ether of N-benzoyl tyramine) from unripe fruit of Aniba riparia (Lauraceae) on the elevated plus maze, open field, rota rod and hole board tests in mice. Riparin I was administered acutely by intraperitoneal (i.p.) and oral routes to male mice at doses of 25 and 50 mg/kg. The results showed that riparin I (25 and 50 mg/kg, i.p. and per os) increased the number of entries and the time of permanence in the open arms in the plus maze test. Similarly, in the hole board test, riparin I in both routes increased the number of head dips. Riparin I with both doses and routes had no effects on spontaneous motor activity in mice or in the rota rod test, but decreased the number of groomings. These results showed that riparin I by both administration routes has effects on the central nervous system with antianxiety effects on the plus maze and hole board tests. The substance is devoid of myorelaxant effects. PMID- 16372364 TI - Assessment of a bioactive compound for its potential antiinflammatory property by tight junction permeability. AB - Lactobacillus probiotic strains are proving to be abundant sources of bioactive components, including antiinflammatory components. Lifree was made of fruits fermented by Lactobacillus paracasei, Lactobacillus reuterrii and Saccharomyces cerevisiae. This study was designed to test these compounds in cell assays measuring epithelial barrier function and proliferation in the first instance. Cell proliferation was measured in mouse fibroblasts cells (3T3NIH) and rat intestinal epithelial cells (IEC-6), and tight junction activity in the kidney epithelial cell line (MDCK). Tight junction permeability was assessed by measuring transepithelial electrical resistance (TER) across confluent monolayers, following the addition of Lifree with or without a challenge with EGTA. Lifree promoted tight junction formation and recovery following loss of TER from challenge with EGTA. On the other hand, Lifree did not stimulate cell growth in either 3T3NIH and IEC-6 cells. Lifree stimulates tight junction maintenance and formation, suggesting it may have potential antiinflammatory properties. PMID- 16372365 TI - Antibacterial activity of polygodial. AB - Polygodial was found to possess moderate antibacterial activity against gram positive bacteria including Bacillus subtilis, Staphylococcus aureus and gram negative bacteria including Escherichia coli and Salmonella choleraesuis with minimum bactericidal concentrations (MBC) of 100 and 100 microg/mL (0.34 mm) and 100 and 50 microg/mL, respectively. The time kill curve study showed that polygodial was bactericidal against B. subtilis and S. choleraesuis. However, there was a difference in its bactericidal action against endospore-forming B. subtilis and food-borne S. choleraesuis. PMID- 16372366 TI - Panax ginseng reduces adriamycin-induced heart failure in rats. AB - The purpose of this study was to investigate the protective effects of Panax ginseng on adriamycin-induced heart failure. Wistar rats were divided into four groups: control, adriamycin, ginseng and adriamycin with ginseng. Adriamycin (cumulative dose, 15 mg/kg) was administered to rats in six equal intraperitoneal injections over a period of 2 weeks. Ginseng was administered via an oral feeding tube once a day for 30 days (cumulative dose, 150 g/kg). At the end of the 5 week post-treatment period, the hearts of the rats were used to study the synthesis rates of DNA, RNA and protein, myocardial antioxidants and lipid peroxidation. At the end of 3 weeks treatment, heart failure was characterized by ascites, congested liver and depressed cardiac function. Nucleic acid as well as protein synthesis was inhibited, lipid peroxidation was increased and myocardial glutathione peroxidase activity was decreased indicating adriamycin-induced heart failure. In contrast, the administration of ginseng, before and concurrent with adriamycin, significantly attenuated the myocardial effects, lowered the mortality as well as the amount of ascites, increased in myocardial glutathione peroxidase, macromolecular biosynthesis and superoxide dismutase activities, with a concomitant decrease in lipid peroxidation. These findings indicated that ginseng may be partially protective against adriamycin-induced heart failure. PMID- 16372367 TI - Analgesic and anticonvulsant properties of Tetrapleura tetraptera (Taub) (Fabaceae) fruit aqueous extract in mice. AB - Previous studies in our laboratories and elsewhere have shown that the fruit of Tetrapleura tetraptera (Taub) (family: Fabaceae) is widely used in African traditional medicine for the management and/or control of an array of human ailments, including schistosomiasis, asthma, epilepsy, hypertension and so on. The present study was designed to investigate the analgesic and anticonvulsant effects of Tetrapleura tetraptera (Taub) fruit aqueous extract (TTE) in mice. Morphine (MPN, 10 mg/kg i.p.), diclofenac (DIC, 100 mg/kg i.p.), phenobarbitone (20 mg/kg i.p.) and diazepam (0.5 mg/kg i.p.) were used, respectively, as reference analgesic and anticonvulsant agents for comparison. T. tetraptera fruit aqueous extract (TTE, 50-800 mg/kg i.p.) produced dose-dependent, significant (p < 0.05-0.001) analgesic effects against thermally and chemically induced pain in mice. Like the standard anticonvulsant agents (phenobarbitone and diazepam) used, T. tetraptera fruit aqueous extract (TTE, 50-800 mg/kg i.p.) significantly (p < 0.05-0.001) delayed the onset of, and antagonized, pentylenetetrazole (PTZ) induced seizures. Aqueous extract of the fruit (TTE, 50-800 mg/kg i.p.) also profoundly antagonized picrotoxin (PCT)-induced seizures, but only partially and weakly antagonized bicuculline (BCL)-induced seizures. However, the results of this experimental animal study indicate that Tetrapleura tetraptera (Taub) fruit aqueous extract (TTE) possesses analgesic and anticonvulsant properties. These findings lend pharmacological support to the suggested folkloric uses of the plant's fruit in the management and/or control of painful, arthritic inflammatory conditions, as well as for the management and/or control of epilepsy and childhood convulsions in some tropical African countries. PMID- 16372368 TI - Protective effect of Spirulina against doxorubicin-induced cardiotoxicity. AB - The generation of reactive oxygen species and mitochondrial dysfunction has been implicated in doxorubicin (DOX)-induced cardiotoxicity. The aim of the present study was to determine whether Spirulina, a blue-green algae, could serve as a cardioprotective agent during DOX treatment in a mouse model. Mice were treated with DOX (4 mg/kg bw, intraperitoneally), weekly, for 4 weeks. Spirulina was administered orally for 3 days twice daily, then for 7 weeks along with the four equal injections of DOX. Cardiotoxicity was assessed, at 3 weeks after the end of the DOX-treatment period, by mortality, volume of ascites, liver congestion, oxidative stress and ultrastructural changes of heart tissue. The DOX-treated animals showed higher mortality (53%) and more ascites. Myocardial damage, as assessed by ultrastructural changes, showed loss of myofibrils, cytoplasmic vacuolization and mitochondrial swelling. Myocardial superoxide dismutase and glutathione peroxidase activities were decreased and lipid peroxidation was increased. Pretreatment with Spirulina significantly protected the mice from DOX induced cardiotoxic effects as evidenced from lower mortality (26%), less ascites, lower levels of lipid peroxidation, normalization of antioxidant enzymes and ultrastructural studies showing minimal damage to the heart. In vitro cytotoxic studies using ovarian cancer cells demonstrated that Spirulina did not compromise the anti-tumor activity of doxorubicin. These results suggest that Spirulina has a protective effect against cardiotoxicity induced by DOX and it may, therefore, improve the therapeutic index of DOX. PMID- 16372369 TI - Gastroprotective activity of sesquiterpene derivatives from Fabiana imbricata. AB - An infusion or decoction of the aerial parts of Fabiana imbricata (Solanaceae) is used in traditional medicine in Chile and Argentina as a digestive and diuretic agent. The main sesquiterpene of the plant was identified as 11-hydroxy-4 amorphen-15-oic acid. At doses of 25, 50 and 100 mg/kg, the compound showed a dose-dependent gastroprotective effect in HCl/EtOH-induced gastric lesions in mice reducing the lesions by 68% at 100 mg/kg. Seven derivatives of the terpene were prepared and their gastroprotective effect was assessed in HCl/EtOH-induced gastric lesions in mice. The cytotoxicity of the products was evaluated in fibroblasts and AGS cells. At 100 mg/kg, 11-hydroxy-4-amorphen-15-p-toluidinamide presented the best gastroprotective effect reducing the gastric lesions by 80%, showing a similar effect to lansoprazole at 20 mg/kg. The compound, however, presented higher cytotoxicity than other derivatives with the IC50 ranging between 110 and 145 microm in AGS cells and fibroblasts, respectively. Most compounds proved to be non-toxic showing cytotoxicity values higher than 1000 microm. The spectroscopic data of six 11-hydroxy-4-amorphen-15-oic acid derivatives are presented here for the first time. PMID- 16372370 TI - Antioxidant and free radical scavenging activities of Misodendrum punctulatum, myzodendrone and structurally related phenols. AB - The peroxyl radical scavenging activity of a dry methanol extract of Misodendrum punctulatum was determined by means of luminol-enhanced chemiluminescence assay, allowing to calculate the total reactive antioxidant potential (TRAP) index equal to 239 +/- 26 microm, expressed in Trolox equivalents. The flavan-3-ol catechin (1) and the phenylbutanone derivative myzodendrone (2) were identified through assay-guided fractionation as active metabolites present in the extract, and their structures were elucidated by chemical and spectroscopic analysis. Three other structurally related synthetic phenols, dehydrozingerone (3), zingerone (4) and myzodendrone aglycone (5), were also analysed using this method. Compounds 1 and 2 were highly effective as free radical scavengers (TRAP = 1257 microm and 1018 microm, respectively) when compared with Trolox (TRAP = 144 microm), used as a standard. Compounds 3 and 5 were also active showing TRAP values of 229 microm and 219 microm, respectively, similarly to that observed for the dry extract. On the other hand, 4 was inactive. Catechin (1) also reduced the production of thiobarbituric acid reactive substances (TBARS) in rat liver homogenates, with IC50 = 26 microg/mL, superior to that obtained for Trolox, IC50 = 73 microg/mL. Compounds 2 and 5 showed IC50 values > 1000 microg/mL, while no activity could be observed for 3, 4, or the extract. PMID- 16372371 TI - Synergistic antiinflammatory effects of pinitol and glucosamine in rats. AB - This study evaluated the antiinflammatory activities of pinitol and glucosamine either alone or in combination against carrageenan- and cotton pellet-induced acute and subacute inflammation in rats. Five groups were included in each of the acute and subacute inflammation studies: the vehicle control group, positive control group (aminopyrine 100 mg/kg), pinitol group (20 mg/kg), glucosamine group (25 mg/kg) and a pinitol (20 mg/kg) and glucosamine (25 mg/kg) combination group. When 20 mg/kg of pinitol was administered to the rats, paw edema induced by the carrageenan injection was significantly suppressed and the level of granuloma formation induced by the cotton pellet implantation was slightly reduced. When 25 mg/kg of glucosamine was administered, paw edema caused by the acute inflammation was slightly reduced and the level of granuloma formation caused by the subacute inflammation was strongly suppressed. Although the combined application of pinitol and glucosamine did not have an additional antiinflammatory effect on the paw edema caused by acute inflammation, it did have an increased antiinflammatory effect on the formation of granuloma induced by subacute inflammation. Therefore, pinitol and glucosamine have an antiinflammatory effect on acute and subacute conditions. Moreover, a synergistic antiinflammatory effect against subacute inflammation was observed when the two chemicals were administered in combination. PMID- 16372372 TI - Immunostimulating activity of pectic polysaccharide from Bergenia crassifolia (L.) Fritsch. AB - The pectic polysaccharide named bergenan BC was obtained using extraction of the green leaves of Siberian tea Bergenia crassifolia (L.) Fritsch. by an aqueous ammonium oxalate. The polysaccharide obtained was proved to comprise mainly d galacturonic acid, galactose, rhamnose, arabinose and glucose residues and appeared to be pectin. Delayed type hypersensitivity (DTH) reaction to aggregated ovalbumin (agOVA) was found to increase in mice that received bergenan solution (2 mg/mL) for 3 weeks. Bergenan BC was observed to enhance the uptake capacity of human neutrophils at a concentration 100 microg/mL and to stimulate the generation of oxygen radicals by mouse peritoneal macrophages in vitro. Bergenan BC was found to increase the spontaneous adhesion of peritoneal leukocytes and failed to influence adhesion stimulated by PMA or adhesion of peritoneal leukocytes incubated in the presence of 5 mm EDTA. Bergenan failed to show cytotoxic action. The viability of peritoneal leukocytes was estimated to be equal to 91% +/- 8% and 90% +/- 7%% in the control and in the pectin solution at a concentration of 1 mg/mL.Thus, bergenan was shown to possess immunostimulating activity in relation to DTH response in vivo and phagocytic activity in vitro. PMID- 16372373 TI - Antidiabetic oligocyclitols in seeds of Mucuna pruriens. AB - Using a combination of chromatographic and NMR techniques, the presence of D chiro-inositol and its two galacto-derivatives is demonstrated in Mucuna pruriens seeds. The quantities detected explain the well-established antiglycaemic effect of Mucuna pruriens seeds. PMID- 16372374 TI - Antiplatelet and antithrombotic activities of methanol extract of Usnea longissima. AB - The antiplatelet and antithrombotic activities of a methanol extract of a medicinal lichen, Usnea longissima, were investigated on platelet aggregation in vitro and on pulmonary thrombosis in vivo. The extract showed concentration dependent inhibitory effects on ADP-induced platelet aggregation, with an IC50 value of 3.6 mg/mL. Using an in vivo mouse thrombotic model in which mice were challenged with an intravenous injection of collagen and epinephrine mixture, oral administration of the extract prior to the injection produced a significant inhibition of thrombotic death or paralysis at 100-200 mg/kg body weight. Aspirin, a representative antiplatelet drug, produced a significant inhibition of thrombotic death at 10-20 mg/kg body weight. The mouse tail bleeding time was significantly prolonged by the addition of the extract. On the other hand, the extract did not show any fibrinolytic activity or alter the coagulation parameters such as activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT) in rat platelets in vitro. These results suggested that the antithrombotic activity of U. longissima extract might be due to antiplatelet activity rather than anticoagulant activity. PMID- 16372375 TI - Effects of Carica papaya (paw paw) seeds extract on the morphology of pituitary gonadal axis of male Wistar rats. AB - The effects of oral administration of Carica papaya seeds extract on the morphology of pituitary, testes and sex accessory glands were studied. The C. papaya extract (50 and 200 mg/kg/day) was administered orally to sexually mature male Wistar rats for 1 and 8 weeks, respectively. The control group received corn oil (vehicle) only. The animals were killed after 1 and 8 weeks treatment for histological preparation. The microscopic examination of the sections of pituitary gonadotrophs (FSH and LH cells) treated with 200 mg/kg of C. papaya extract showed pronounced hypertrophy, while section of rats treated with 50 mg/kg showed mild hypertrophy and hyperplasia. Whereas the testes of rats treated with C. papaya extract at 50 and 200 mg/kg revealed gradual degeneration of germ cells, Sertoli cells and Leydig cells as well as germinal epithelium. However, tubules of epididymes of rats treated with extract, 200 mg/kg, appeared empty indicating the degeneration of sperm cells in the lumina. The sections of prostate glands of rats treated with 200 mg/kg extract showed coagulation of secretion in the lumina as well as empty tubules with cell debris, while the effect was reduced in rats treated with 50 mg/kg. The seminal vesicles showed progressive collapse and shrinkage of villi. These results suggest that C. papaya extract interfered with the pituitary - gonadal axis to influence male reproductive functions, which confirmed its antifertility property as reported previously. PMID- 16372377 TI - Phytochemical investigation and antioxidant activity of Duranta repens. AB - The ethylacetate fraction of the methanol extract of Duranta repens L. (Verbenaceae) showed radicalscavenging activity in the 1,1-diphenyl-2 picrylhydrazyl (DPPH) assay. Three compounds were isolated from this fraction, i.e. the phenylethanoid glycoside acteoside, the iridoid lamiide and the saponin pseudo-ginsenoside-RT1. Acteoside showed an IC50 of 3.05 +/- 0.09 microg/mL in the DPPH assay, while lamiide and pseudo-ginsenoside-RT1 were not active. PMID- 16372376 TI - Antiviral properties of ent-labdene diterpenes of Andrographis paniculata nees, inhibitors of herpes simplex virus type 1. AB - Andrographolide, neoandrographolide and 14-deoxy-11,12-didehydroandrographolide, ent-labdene diterpenes isolated from Andrographis paniculata showed viricidal activity against herpes simplex virus 1 (HSV-1). None of these compounds exhibited significant cytotoxicity at viricidal concentrations. PMID- 16372378 TI - Antioxidant activity of Plantago bellardii All. AB - The aim of this study was to evaluate the in vitro antioxidant activity of the methanol extract of Plantago bellardii All. aerial parts. This was assessed by two different tests, scavenging of 1,1-diphenyl-2-picrylhydrazil (DPPH) radical, and inhibition of lipid peroxidation on liposomes prepared from bovine brain extract. In both tests the extract showed a potent antioxidant effect. The characterization of the major compounds in the extract as rutin, geniposide and verbascoside was performed by isolation and HPLC comparison with authentic samples. They were quantified by HPLC for the flavonoids and colorimetry for iridoids. The compounds that contribute most to the antioxidant activity were shown to be verbascoside and rutin. PMID- 16372379 TI - Effect of Alkanna albugam root on LDL oxidation. A comparative study with species of the Lamiaceae family. AB - The role of natural products in preventing diseases, such as atherosclerosis, is being recognized increasingly. In this study, the in vitro antioxidant activity of the polar extract obtained from Alkanna albugam root, belonging to the Boraginaceae family, against LDL oxidation, was investigated and compared with the respective activity of Greek Mentha piperita, Rosmarinus officinalis and Sideritis clandestina, belonging to the Lamiaceae family. Of the extracts tested, only those from Alkanna albugam root and Mentha piperita exhibited statistically significant activity. Weaker activity was shown by the extracts obtained from Rosmarinus officinalis and Sideritis clandestina. PMID- 16372380 TI - The effect of ciprofloxacin and clarithromycin on sildenafil oral bioavailability in human volunteers. AB - Sildenafil is the first oral therapeutic agent for the management of male erectile dysfunction. Its oral bioavailability is only 40% due to extensive presystemic elimination, mainly by CYP3A4. This study examined the effect of coadministration of ciprofloxacin or clarithromycin, which inhibit CYP3A4, on the bioavailability and pharmacokinetics of sildenafil. Twelve healthy male volunteers received sildenafil alone or after pretreatment with the inhibitors in a balanced three-way crossover design. The pharmacokinetic analysis showed that ciprofloxacin coadministration with sildenafil significantly increased the AUC from 1407 +/- 380 to 2986 +/- 917 microg h/l (90% confidence interval 119%-159%) and the Cmax from 287 +/- 67 to 623 +/- 192 microg/l (90% confidence interval 127%-152%). Similarly, clarithromycin coadministration increased sildenafil AUC from 1407 +/- 380 to 3209 +/- 762 microg h/l (90% confidence interval 127%-161%) and Cmax from 287 +/- 67 to 694 +/- 259 microg/l (90% confidence interval 132% 157%). Ciprofloxacin coadministration and clarithromycin coadministration with sildenafil did not affect the rate of sildenafil absorption significantly. These results indicate that coadministration of ciprofloxacin and clarithromycin significantly increased sildenafil bioavailability which can be attributed to the inhibitory effect of ciprofloxacin and clarithromycin on CYP3A4. Dose adjustment of sildenafil is thus necessary when administered with such drugs. PMID- 16372381 TI - Differentiation of fluoronitroaniline isomers by negative-ion electrospray mass spectrometry. AB - Tetra- and trifluoronitroanilines were studied by electrospray ionization mass spectrometry. These compounds gave signals only in the negative-ion mode. It was found that the so-called 'in-source' fragmentation, induced by cone voltage increase, enables differentiation of isomers. For para-nitroanilines, in contrast to ortho derivatives, the loss of NO(2) was the most favored process and other fragment ions were characterized by low abundances. For trifluoro conjugates the substitution pattern of aromatic ring by fluorine atoms also affected their fragmentation patterns. For example, in 2,3,6-trifluoro-4-nitroaniline, in contrast to 2,3,5-trifluoro-4-nitroaniline, efficient NO loss, followed by HF loss, took place. PMID- 16372382 TI - Characterization of N-glycans of recombinant human thyrotropin using mass spectrometry. AB - Thyroid-stimulating hormone is a vital component of the regulatory mechanism that maintains the structure and function of the thyroid gland and governs thyroid hormone release. In this paper we report the first detailed structural characterization of the N-linked oligosaccharides of recombinant human thyroid stimulating hormone (rhTSH). Using a strategy combining mass spectrometric analysis and sequential exoglycosidase digestion, we have defined the structures of the N-glycans released from recombinant human thyrotropin by peptide N glycosidase F. All glycans are complex-type glycans and are mainly of the bi- and triantennary type with variable degrees of fucosylation and sialylation. The major non-reducing epitope in the complex-type glycans is: NeuAcalpha2-3Galbeta1 4GlcNAc (sialylated LacNAc). The carbohydrate microheterogeneity at the three glycosylation sites was studied using reversed-phase high-performance liquid chromatography (RP-HPLC), concanavalin A affinity chromatography and mass spectrometric techniques, including both matrix-assisted laser desorption/ionization (MALDI) and electrospray. rhTSH was reduced, carboxymethylated and then digested with trypsin. The mixture of peptides and glycopeptides was subjected to RP-HPLC and the structures of the glycopeptides were determined by MALDI in conjunction with on-target exoglycosidase digestions. After PNGase F digestion, the peptide moiety of the glycopeptide was determined by the presence of the b- and y-series ions derived from its amino acid sequence in the quadrupole time-of-flight tandem mass (QTOF-MS/MS) spectrum. Glycosylation sites Asn-alpha52 and Asn-alpha78 contain mainly bi- and triantennary complex type glycans. Only glycosylation site Asn-alpha52 bears fucosylated N-glycans. Minor tetraantennary complex structures were also observed on both glycosylation sites. Profiling of the carbohydrate moieties of Asn-beta23 indicates a large heterogeneity. Bi-, tri-, and tetraantennary N-glycans were present at this site. These data demonstrate site-specificity of glycosylation in the alpha subunit but not in the beta subunit of rhTSH with Asn-alpha52 bearing essentially di- and triantennary glycans with or without core fucosylation and bi- and triantennary glycans with no core fucosylation being attached to Asn-alpha78. PMID- 16372383 TI - Static secondary ion mass spectrometry for nanoscale analysis: surface characterisation of electrospun nanofibres. AB - The viability of static secondary ion mass spectrometry (S-SIMS) for selected applications of nanoscale analysis has been investigated, focusing on nanofibres produced by electrospinning (ES) as a test case. The samples consist of non-woven nanowebs of which the individual fibres have diameters in the range of 100 nm. Use of solutions with functionalised polymers or polar additives potentially allows the surface composition to be tailored as a function of the application. So far nanowebs are primarily characterised by morphological examination. This paper describes the first detailed characterisation of molecular composition at the surface of nanofibres electrospun from poly(epsilon-caprolactone) (PCL) solutions in acetone containing 0-15 mol% (relative to PCL) of cetyltrimethylammonium bromide (CTAB). Application of S-SIMS to nanowebs has allowed mass spectra to be recorded containing the major diagnostic ions of both components. Their relative intensities point to surface enrichment and depletion of the polar CTAB additive relative to the PCL matrix for samples electrospun from solution containing low and high CTAB concentrations, respectively. PMID- 16372384 TI - Identification of an unusual naturally occurring apolar fatty acid amide in mammalian brain and a method for its quantitative determination. AB - Fatty acid amides (FAAs), such as the N-acylamides, N-acylethanolamides, N acyldopamines and N-acylamino acids, are now emerging as an important new class of lipid-signalling molecules. This paper provides evidence, based on high performance liquid chromatography/electrospray ionisation mass spectrometry (HPLC/ESI-MS/MS), gas chromatography/mass spectrometry (GC/MS) and 1H-NMR, of the occurrence in mouse and bovine brain extracts of a compound characterised by a mass spectrum attributable to a FAA not previously described, namely, the isopropyl-amide of stearic acid (SIPA). A highly sensitive GC/MS method was developed for quantification of naturally occurring SIPA and, also, for purposes of comparison, that of palmitoylethanolamide (PEA), a structurally related compound commonly determined in animal tissues. The results obtained show that SIPA levels in mouse brain are 8-10-fold higher than those of PEA. Moreover, SIPA was found in human neuroblastoma cell (SHSY-5Y) extracts, at significantly higher levels following exposure of the cells to the mitochondrial inhibitor rotenone. All this evidence not only shows surprisingly that SIPA may be found naturally in mammalian biological extracts despite the unusual functional group (i.e. isopropylamide) implicated, but also raises many important questions concerning its biological origin. PMID- 16372385 TI - Validation of a gas chromatography/triple quadrupole mass spectrometry based method for the quantification of pesticides in food commodities. AB - A new multiresidue method has been validated in cucumber matrix for the routine analysis of 130 multiclass pesticide residues by gas chromatography/triple quadrupole mass spectrometry. The pesticides were extracted with ethyl acetate. A first identification of the pesticides was based on a tandem mass spectrometric (MS/MS) screening method, which monitors a single transition for each target compound, in less than 12 min. After that, potentially non-negative samples were analyzed again by the MS/MS confirmation/quantification method, which monitors two or three MS/MS transitions for each compound, also in less than 12 min. Performance characteristics, such as trueness, precision, linear range, detection limit (LOD) and quantification limit (LOQ), for each pesticide were calculated. The average recoveries obtained ranged between 70 and 120% at three different fortification levels (25, 200 and 500 microg/kg) with precision, expressed as relative standard deviation (RSD), values lower than 15%. The calculated LOD and LOQ were typically <3.2 and 9.6 microg/kg, respectively. Such limits were much lower than the maximum residue levels (MRLs) established by European legislation. The proposed methodology was applied to the determination of pesticides in real vegetable samples from Almeria (Spain). PMID- 16372386 TI - Semi-parametric accelerated failure time regression analysis with application to interval-censored HIV/AIDS data. AB - This paper demonstrates a way to investigate a potentially non-linear relationship between an interval-censored response variable and a continuously distributed explanatory variable. A potentially non-linear effect of a continuous explanatory variable on the response is incorporated into an accelerated failure time model, forming a partial linear model. A sieve maximum likelihood estimator (MLE) is suggested to simultaneously estimate all the parameters. The sieve MLE is shown to be asymptotically efficient and normally distributed. Simulation studies show that the proposed estimators for the scale and regression parameters are robust and efficient, and the estimator for the non-linear function is able to capture the shape of a variety of smooth non-linear functions. The model is applied to observational HIV data, where the response variable is the time to suppression of HIV viral load after initiation of antiretroviral therapy, and baseline viral load is investigated as a potentially non-linear effect. PMID- 16372387 TI - A hierarchical non-homogenous Poisson model for meta-analysis of adenoma counts. AB - We use a hierarchical model for a meta-analysis that combines information from autopsy studies of adenoma prevalence and counts. The studies we included reported findings using a variety of adenoma prevalence groupings and age categories. We use a non-homogeneous Poisson model for multinomial bin probabilities. The Poisson model allows risk to depend on age and sex, and incorporates extra-Poisson variability. We evaluate model fit using the posterior predicted distribution of adenoma prevalence reported by the studies included in our analyses and validate our model using adenoma prevalence reported by more recent colonoscopy studies. For 1990, the estimated adenoma prevalence among Americans at age 60 is 40.3 per cent for men compared to 29.2 per cent for women. PMID- 16372388 TI - Functional ANOVA with random functional effects: an application to event-related potentials modelling for electroencephalograms analysis. AB - The differential effects of basic visual or auditory stimuli on electroencephalograms (EEG), named event related potentials (ERPs), are often used to evaluate the impact of treatments on brain performances. In the present paper, we propose a P-splines based model that can be used to evaluate treatment effect on the timing and the amplitude of some peaks of the ERPs curves. Functional ANOVA is an adaptation of linear model or analysis of variance to analyse functional observations. The changes in the functional of interest effects are generally described using smoothing splines. Eilers and Marx proposed to work with P-splines, a combination of B-splines and difference penalties on coefficients. We define a P-splines model for ERPs curves combined with random effects. In particular, we show that it is a useful alternative to classical strategies requiring the visual and usually imprecise localization of specific ERP peaks from curves with a low signal-to-noise ratio. PMID- 16372389 TI - Sample size estimation for the van Elteren test--a stratified Wilcoxon-Mann Whitney test. AB - The van Elteren test is a type of stratified Wilcoxon-Mann-Whitney test for comparing two treatments accounting for strata. In this paper, we study sample size estimation methods for the asymptotic version of the van Elteren test, assuming that the stratum fractions (ratios of each stratum size to the total sample size) and the treatment fractions (ratios of each treatment size to the stratum size) are known in the study design. In particular, we develop three large-sample sample size estimation methods and present a real data example to illustrate the necessary information in the study design phase in order to apply the methods. Simulation studies are conducted to compare the performance of the methods and recommendations are made for method choice. Finally, sample size estimation for the van Elteren test when the stratum fractions are unknown is also discussed. PMID- 16372390 TI - Estimation of genetic and environmental factors for melanoma onset using population-based family data. AB - Estimation of genetic and environmental contributions to cancers falls in the framework of generalized linear mixed modelling with several random effect components. Computational challenges remain, however, in dealing with binary or survival phenotypes. In this paper, we consider the analysis of melanoma onset in a population of 2.6 million nuclear families in Sweden, for which none of the current survival-based methodologies is feasible. We treat the disease outcome as a binary phenotype, so that the standard proportional hazard model leads to a generalized linear model with the complementary-log link function. For rare diseases this link is very close to the probit link, and thus allows the use of marginal likelihood for the estimation of the variance components. We correct for the survival length bias by censoring the parent generation within each family at the time they attain the same cumulative hazard as the child generation, thus improving the validity of the estimates. Our finding that childhood shared environment in addition to genetic factors had a considerable effect on the development of melanoma is consistent with epidemiological studies. PMID- 16372391 TI - Sample sizes for self-controlled case series studies. AB - We derive several formulae for the sample size required for a study designed using the self-controlled case series method without age effects. We investigate these formulae by simulation, and identify one based on the signed root likelihood ratio statistic which performs well. We extend this method to allow for age effects, which can have a big impact on the sample size needed. This more general sample size formula is also found to perform well in a broad range of situations. PMID- 16372392 TI - Minimally invasive surgery of the thyroid and parathyroid glands. PMID- 16372393 TI - Dietary fibre. PMID- 16372394 TI - Apoptosis, ageing and redox homeostasis in yeasts. PMID- 16372395 TI - 10th EASD/JDRF Oxford Workshop. What is a beta-cell and can we improve it? PMID- 16372396 TI - Acute compartment syndrome of the limb. PMID- 16372397 TI - Psychometric properties of the Grandparent Perceptions of Family Scale (GPFS). PMID- 16372398 TI - Visits to the emergency department as transactional data. AB - Patients arrive at the hospital emergency department for treatment on a random basis. The amount of time required for treatment is a function of the triage level, the patient diagnosis, and the congestion that exists in the emergency department (ED) at the time of patient arrival. The implementation of electronic medical records in the ED permits the accurate tracking and examination of time to allow for improved scheduling of personnel and for the development of protocols for diagnoses that occur on a daily basis in the ED. The SAS Institute in Cary, North Carolina, has developed a method called High Performance Forecasting System that allows for the prediction of time series with random time points. The target variable is the amount of time needed to treat individual patients from the time they enter the system through triage to the time they are discharged. Variability in treatment time by ED personnel can also be examined. Results indicate that there is a "cut point" during the day when the time needed to treat each patient increases. In addition, some specific diagnoses can be anticipated on a regular basis, and the amount of time needed on a daily basis can be predicted by using the forecasting system. Personnel can be scheduled in accordance with the peak treatment time. PMID- 16372399 TI - Brain development. PMID- 16372401 TI - Sustained release of PEG-g-chitosan complexed DNA from poly(lactide-co glycolide). AB - Chitosan and its derivatives have emerged as promising gene-delivery vehicles because of their capability to form polyplexes with plasmid DNA and enhance its transport across cellular membranes through endocytosis. Evidently, polyplexes of chitosan and DNA significantly improve transfection efficiency; however, these polyplexes are not capable of sustained DNA release and, thus, prolong gene transfer. In order to achieve prolonged delivery of DNA/chitosan polyplexes, we have formulated microspheres by physically combining poly(ethylene glycol) grafted chitosan (PEG-g-CHN) with poly(lactide-co-glycolide) (PLGA) using a modified conventional in-emulsion solvent evaporation method. Electrophoretic analysis of materials released from these microspheres suggests the presence of PEG-g-CHN complexed DNA and these microspheres are capable of sustained release of DNA/PEG-g-CHN for at least 9 weeks. The rate of DNA release can be modulated by varying the amount of PEG-g-CHN. The release products from these microspheres are bioactive and show enhanced transfection in vitro over DNA released from conventional PLGA microspheres containing no PEG-g-CHN. In vivo experiments also show that these microspheres are capable of achieving gene transfer in a rat hind limb muscle model. PMID- 16372402 TI - The stem cell continuum: a new model of stem cell regulation. AB - Most models of hematopoiesis have been hierarchical in nature. This is based on a large volume of correlative data. Recent work has indicated that, at least at the stem/progenitor level, hematopoiesis may, in fact, be a continuum of transcriptional opportunity. The most primitive hematopoietic stem cells are either continually cycling at a slow rate or entering and exiting cell cycle. Associated with this cycle passage are changes in functional phenotype including reversible alterations in engraftment, adhesion protein expression, cytokine receptor expression, homing to marrow, and progenitor cell numbers. Global gene expression, as measured in one point in cycle, is also markedly altered. The differentiation potential of the marrow as it transits cell cycle in response to a set differentiation stimulus also shows marked variations. This cycle-related plasticity has been clearly established for hematopoiesis. It also holds for the ability of murine marrow stem cells to home to lung and to convert to pulmonary cells. These data indicate that bone marrow stem cells can probably not be defined as discrete entities but must rather be studied on a population basis. They also indicate that mathematical modeling will become progressively more important in this field. PMID- 16372400 TI - Cancer. PMID- 16372403 TI - Small molecules that delay S phase suppress a zebrafish bmyb mutant. AB - Bmyb is a ubiquitously expressed transcription factor involved in cellular proliferation and cancer. Loss of bmyb function in the zebrafish mutant crash&burn (crb) results in decreased cyclin B1 expression, mitotic arrest and genome instability. These phenotypic observations in crb mutants could be attributed to the decreased expression of cyclin B1, a cell-cycle regulatory protein that is responsible for driving cell progression from G2 through mitosis. To identify small molecules that interact with the bmyb pathway, we developed an embryo-based suppressor screening strategy. In 16 weeks we screened a diverse approximately 16,000 compound library, and discovered one previously unknown compound, persynthamide (psy, 1), that suppressed bmyb-dependent mitotic defects. Psy-treated embryos showed an S-phase delay, and knockdown of the cell-cycle checkpoint regulator ataxia telangiectasia--and Rad-related kinase (ATR) abrogated the suppression of crb. The DNA synthesis inhibitors aphidicolin (2) and hydroxyurea (3) also suppressed crb. S-phase inhibition upregulated cyclin B1 mRNA, promoting the progression of cells through mitosis. Our study demonstrates that chemical suppressor screening in zebrafish can identify compounds with cell cycle activity and can be used to identify pathways that interact with specific cell-cycle phenotypes. PMID- 16372405 TI - Alzheimer's disease cerebrospinal fluid biomarker discovery: a proteomics approach. AB - There is significant interest in the identification of effective biomarkers for Alzheimer's disease. Such biomarkers could aid in the clinical diagnosis of the disease and may be useful in assessing the efficacy of various treatment strategies. The search for biomarkers often includes the analysis of changes in cerebrospinal fluid protein expression that correlate with disease. These changes can be measured using a variety of technologies for protein expression profiling. Although there is great promise in the application of these methods to biomarker discovery based on some preliminary observations, there are significant issues in the capabilities of most of these technologies that have limited their effective application. The most recent literature involving proteomic discovery of new cerebrospinal fluid biomarkers for Alzheimer's disease is reviewed. PMID- 16372404 TI - Actin is the primary cellular receptor of bistramide A. AB - Bistramide A (1) is a marine natural product with broad, potent antiproliferative effects. Bistramide A has been reported to selectively activate protein kinase C (PKC) delta, leading to the view that PKCdelta is the principal mediator of antiproliferative activity of this natural product. Contrary to this observation, we established that bistramide A binds PKCdelta with low affinity, does not activate this kinase in vitro and does not translocate GFP-PKCdelta. Furthermore, we identified actin as the cellular receptor of bistramide A. We report that bistramide A disrupts the actin cytoskeleton, inhibits actin polymerization, depolymerizes filamentous F-actin in vitro and binds directly to monomeric G actin in a 1:1 ratio with a Kd of 7 nM. We also constructed a fully synthetic9 bistramide A-based affinity matrix and isolated actin as a specific bistramide A binding protein. This activity provides a molecular explanation for the potent antiproliferative effects of bistramide A, identifying it as a new biochemical tool for studies of the actin cytoskeleton and as a potential lead for development of a new class of antitumor agents. PMID- 16372406 TI - Integrating genomics and transcriptomics with geo-ethnicity and the environment for the resolution of complex cardiovascular diseases. AB - One of the crucial steps on the way to individualized medicine to treat cardiovascular disease (CVD) is to better understand the identities, roles, extent and at least the major patterns of interaction between influential genomic and environmental factors. It is clear that such a bold goal can hardly be achieved without a major upgrade of our conceptualization of the phenomena studied, taking advantage of recent developments of novel technological and computational tools. Firstly, the search for the genomic components of the most common multifactorial CVDs is no longer restricted to protein-coding genes; truly genome-wide investigations should replace them in both humans and animal models. Secondly, the 'environment' has also undergone semantic expansion, incorporating such remote constituents as developmental plasticity and epigenetics on one side, and socioeconomic status on the other. To elucidate and analyze the resulting complex picture, appropriate statistical models and approaches need to be designed to tackle issues such as population stratification and admixture, multiple testing, and multidimensionality reduction in models involving multiple genes and environmental factors. Eventually, an integrated platform bringing together all of the above will probably be necessary to secure relevant information specific to a particular combination of conditions and settings (age, geo-ethnicity and exposure), which may perhaps become visible only after a step back, through systems (network) biology. PMID- 16372407 TI - Developing new agents for the treatment of childhood cancer. AB - The development of new drugs for the treatment of childhood cancer presents novel challenges, and pediatric clinical trials of a new agent are often initiated many years after testing in adults. It is estimated that there are over 400 new entities being developed as cancer treatments, although few of these have been developed specifically for the treatment of childhood malignancies. This raises the question of how agents can be prioritized for pediatric clinical testing. In this review, the molecular characteristics of childhood cancers that may be valuable in steering choices for rational or molecularly targeted treatments are described, and the Pediatric Preclinical Testing Program, a National Cancer Institute initiative to identify agents that should be prioritized for clinical evaluation against childhood cancer, is presented. PMID- 16372408 TI - Treatment delivery software for a new clinical grade ultrasound system for thermoradiotherapy. AB - A detailed description of a clinical grade Scanning Ultrasound Reflector Linear Array System (SURLAS) applicator was given in a previous paper [Med. Phys. 32, 230-240 (2005)]. In this paper we concentrate on the design, development, and testing of the personal computer (PC) based treatment delivery software that runs the therapy system. The SURLAS requires the coordinated interaction between the therapy applicator and several peripheral devices for its proper and safe operation. One of the most important tasks was the coordination of the input power sequences for the elements of two parallel opposed ultrasound arrays (eight 1.5 cm x 2 cm elements/array, array 1 and 2 operate at 1.9 and 4.9 MHz, respectively) in coordination with the position of a dual-face scanning acoustic reflector. To achieve this, the treatment delivery software can divide the applicator's treatment window in up to 64 sectors (minimum size of 2 cm x 2 cm), and control the power to each sector independently by adjusting the power output levels from the channels of a 16-channel radio-frequency generator. The software coordinates the generator outputs with the position of the reflector as it scans back and forth between the arrays. Individual sector control and dual frequency operation allows the SURLAS to adjust power deposition in three dimensions to superficial targets coupled to its treatment window. The treatment delivery software also monitors and logs several parameters such as temperatures acquired using a 16-channel thermocouple thermometry unit. Safety (in particular to patients) was the paramount concern and design criterion. Failure mode and effects analysis (FMEA) was applied to the applicator as well as to the entire therapy system in order to identify safety issues and rank their relative importance. This analysis led to the implementation of several safety mechanisms and a software structure where each device communicates with the controlling PC independently of the others. In case of a malfunction in any part of the system or a violation of a user-defined safety criterion based on temperature readings, the software terminates treatment immediately and the user is notified. The software development process consisting of problem analysis, design, implementation, and testing is presented in this paper. Once the software was finished and integrated with the hardware, the therapy system was extensively tested. Results demonstrated that the software operates the SURLAS as intended with minimum risk to future patients. PMID- 16372409 TI - Dose rate constant of a cesium-131 interstitial brachytherapy seed measured by thermoluminescent dosimetry and gamma-ray spectrometry. AB - The aim of this work was to conduct an independent determination of the dose rate constant of the newly introduced Model CS-1 131Cs seed. A total of eight 131Cs seeds were obtained from the seed manufacturer. The air-kerma strength of each seed was measured by the manufacturer whose calibration is traceable to the air kerma strength standard established for the 131Cs seeds at the National Institute of Standards and Technology (1 sigma uncertainty < 1%). The dose rate constant of each seed was measured by two independent methods: One based on the actual photon energy spectrum emitted by the seed using gamma-ray spectrometry and the other based on the dose-rate measured by thermoluminescent dosimeter (TLD) in a Solid Water phantom. The dose rate constant in water determined by the gamma-ray spectrometry technique and by the TLD dosimetry are 1.066 +/- 0.064 cGyh(-1)U(-1) and 1.058 +/- 0.106 cGyh(-1)U(-1), respectively, showing excellent agreement with each other. These values, however, are approximately 15% greater than a previously reported value of 0.915 cGyh(-1)U(-1) [Med. Phys. 31, 1529-1538 (2004)]. Although low-energy fluorescent x rays at 16.6 and 18.7 keV, originating from niobium present in the seed construction, were measured in the energy spectrum of the 131Cs seeds, their yields were not sufficient to lower the dose rate constant to the value of 0.915 cGyh(-1)U(-1). Additional determinations of the dose rate constant may be needed to establish an AAPM recommended consensus value for routine clinical use of the 131Cs seed. PMID- 16372410 TI - Reduction of motion blurring artifacts using respiratory gated CT in sinogram space: a quantitative evaluation. AB - Techniques have been developed for reducing motion blurring artifacts by using respiratory gated computed tomography (CT) in sinogram space and quantitatively evaluating the artifact reduction. A synthetic sinogram was built from multiple scans intercepting a respiratory gating window. A gated CT image was then reconstructed using the filtered back-projection algorithm. Wedge phantoms, developed for quantifying the motion artifact reduction, were scanned while being moved using a computer-controlled linear stage. The resulting artifacts appeared between the high and low density regions as an apparent feature with a Hounsfield value that was the average of the two regions. A CT profile through these regions was fit using two error functions, each modeling the partial-volume averaging characteristics for the unmoving phantom. The motion artifact was quantified by determining the apparent distance between the two functions. The blurring artifact had a linear relationship with both the speed and the tangent of the wedge angles. When gating was employed, the blurring artifact was reduced systematically at the air-phantom interface. The gated image of phantoms moving at 20 mm/s showed similar blurring artifacts as the nongated image of phantoms moving at 10 mm/s. Nine patients were also scanned using the synchronized respiratory motion technique. Image artifacts were evaluated in the diaphragm, where high contrast interfaces intercepted the imaging plane. For patients, this respiratory gating technique reduced the blurring artifacts by 9%-41% at the lung diaphragm interface. PMID- 16372411 TI - Exact BPF and FBP algorithms for nonstandard saddle curves. AB - A hot topic in cone-beam CT research is exact cone-beam reconstruction from a general scanning trajectory. Particularly, a nonstandard saddle curve attracts attention, as this construct allows the continuous periodic scanning of a volume of-interest (VOI). Here we evaluate two algorithms for reconstruction from data collected along a nonstandard saddle curve, which are in the filtered backprojection (FBP) and backprojection filtration (BPF) formats, respectively. Both the algorithms are implemented in a chord-based coordinate system. Then, a rebinning procedure is utilized to transform the reconstructed results into the natural coordinate system. The simulation results demonstrate that the FBP algorithm produces better image quality than the BPF algorithm, while both the algorithms exhibit similar noise characteristics. PMID- 16372412 TI - Effect of correlation on combining diagnostic information from two images of the same patient. AB - We have shown previously, in the context of computer-aided diagnosis (CAD), that information derived from multiple images of the same patient can be used to improve diagnostic performance. In that work, we ignored the correlation among multiple images of the same patient. In the present study, we investigate theoretically, within the framework of receiver operating characteristic (ROC) analysis, the effect of correlation on three methods for combining quantitative diagnostic information from two images: taking the average, the maximum, and the minimum of a pair of normally distributed decision variables. We assume, as in our previous work, that the quantitative diagnostic information obtained from the two images of a given patient can be transformed monotonically to two latent decision variables that are normally distributed. Similar to the situation of uncorrelated images, we found that (1) the average always improves the area under the ROC curve (AUC) compared to the single-view image; (2) the maximum and the minimum can also, but not always, improve the AUC; and (3) each method can be the best method in certain situations. In addition, as the correlation strength increases, the average performs the best less often, whereas the maximum and the minimum perform the best more often. These theoretical results are illustrated with analysis of a mammography study. PMID- 16372413 TI - 3D MRI-based tumor delineation of ocular melanoma and its comparison with conventional techniques. AB - The aim of this study is to (1) compare the delineation of the tumor volume for ocular melanoma on high-resolution three-dimensional (3D) T2-weighted fast spin echo magnetic resonance imaging (MRI) images with conventional techniques of A- and B-scan ultrasound, transcleral illumination, and placement of tantalum markers around tumor base and (2) to evaluate whether the surgically placed marker ring tumor delineation can be replaced by 3D MRI based tumor delineation. High-resolution 3D T2-weighted fast spin echo (3D FSE) MRI scans were obtained for 60 consecutive ocular melanoma patients using a 1.5 T MRI (GE Medical Systems, Milwaukee, WI), in a standard head coil. These patients were subsequently treated with proton beam therapy at the UC Davis Cyclotron, Davis, CA. The tumor was delineated by placement of tantalum rings (radio-opaque markers) around the tumor periphery as defined by pupillary transillumination during surgery. A point light source, placed against the sclera, was also used to confirm ring agreement with indirect ophthalmoscopy. When necessary, intraoperative ultrasound was also performed. The patients were planned using EYEPLAN software and the tumor volumes were obtained. For analysis, the tumors were divided into four categories based on tumor height and basal diameter. In order to assess the impact of high-resolution 3D T2 FSE MRI, the tumor volumes were outlined on the MRI scans by two independent observers and the tumor volumes calculated for each patient. Six (10%) of 60 patients had tumors, which were not visible on 3D MRI images. These six patients had tumors with tumor heights < or = 3 mm. A small intraobserver variation with a mean of (-0.22 +/- 4)% was seen in tumor volumes delineated by 3D T2 FSE MR images. The ratio of tumor volumes measured on MRI to EYEPLAN for the largest to the smallest tumor volumes varied between 0.993 and 1.02 for 54 patients. The tumor volumes measured directly on 3D T2 FSE MRI ranged from 4.03 to 0.075 cm3. with a mean of 0.87 +/- 0.84 cm3. The tumor shapes obtained from 3D T2 FSE MR images were comparable to the tumor shapes obtained using EYEPLAN software. The demonstration of intraocular tumor volumes with the high-resolution 3D fast spin echo T2 weighted MRI is excellent and provides additional information on tumor shape. We found a high degree of accuracy for tumor volumes with direct MRI volumetric measurements in uveal melanoma patients. In some patients with extra large tumors, the tumor base and shape was modified, because of the additional information obtained from 3D T2 FSE MR images. PMID- 16372414 TI - An alternative solution to the nonuniform noise propagation problem in fan-beam FBP image reconstruction. AB - It has been observed that the variances in reconstructed images from stationary noisy projection data by fan-beam filtered backprojection (fFBP) algorithm with ramp in the filtering step and linear interpolation in the backprojection step are nonuniform across the field-of-view. This is believed to be caused by the distance-dependent 1/L2 factor in the fFBP reconstruction formula. Shift-variant filtration approach in the filtering step has been investigated to address the nonuniform noise propagation problem in fFBP. In this work, we present an alternative solution by the use of spatially variant weighting, instead of spatially invariant interpolation, in the backprojection step, while maintaining the homogeneous ramp filtering. Both theoretically predicted and empirically determined results concur and demonstrate the effectiveness of the presented alternative solution. PMID- 16372415 TI - Dual-energy digital mammography for calcification imaging: scatter and nonuniformity corrections. AB - Mammographic images of small calcifications, which are often the earliest signs of breast cancer, can be obscured by overlapping fibroglandular tissue. We have developed and implemented a dual-energy digital mammography (DEDM) technique for calcification imaging under full-field imaging conditions using a commercially available aSi:H/CsI:Tl flat-panel based digital mammography system. The low- and high-energy images were combined using a nonlinear mapping function to cancel the tissue structures and generate the dual-energy (DE) calcification images. The total entrance-skin exposure and mean-glandular dose from the low- and high energy images were constrained so that they were similar to screening-examination levels. To evaluate the DE calcification image, we designed a phantom using calcium carbonate crystals to simulate calcifications of various sizes (212-425 microm) overlaid with breast-tissue-equivalent material 5 cm thick with a continuously varying glandular-tissue ratio from 0% to 100%. We report on the effects of scatter radiation and nonuniformity in x-ray intensity and detector response on the DE calcification images. The nonuniformity was corrected by normalizing the low- and high-energy images with full-field reference images. Correction of scatter in the low- and high-energy images significantly reduced the background signal in the DE calcification image. Under the current implementation of DEDM, utilizing the mammography system and dose level tested, calcifications in the 300-355 microm size range were clearly visible in DE calcification images. Calcification threshold sizes decreased to the 250-280 microm size range when the visibility criteria were lowered to barely visible. Calcifications smaller than approximately 250 microm were usually not visible in most cases. The visibility of calcifications with our DEDM imaging technique was limited by quantum noise, not system noise. PMID- 16372416 TI - Exact fan-beam and 4pi-acquisition cone-beam SPECT algorithms with uniform attenuation correction. AB - This paper presents analytical fan-beam and cone-beam reconstruction algorithms that compensate for uniform attenuation in single photon emission computed tomography. First, a fan-beam algorithm is developed by obtaining a relationship between the two-dimensional (2D) Fourier transform of parallel-beam projections and fan-beam projections. Using this relationship, 2D Fourier transforms of equivalent parallel-beam projection data are obtained from the fan-beam projection data. Then a quasioptimal analytical reconstruction algorithm for uniformly attenuated Radon data, developed by Metz and Pan, is used to reconstruct the image. A cone-beam algorithm is developed by extending the fan beam algorithm to 4pi solid angle geometry. The cone-beam algorithm is also an exact algorithm. PMID- 16372417 TI - Intra- and interfractional patient motion for a variety of immobilization devices. AB - The magnitude of inter- and intrafractional patient motion has been assessed for a broad set of immobilization devices. Data was analyzed for the three ordinal directions--left-right (x), sup-inf (y), and ant-post (z)--and the combined spatial displacement. We have defined "rigid" and "non-rigid" immobilization devices depending on whether they could be rigidly and reproducibly connected to the treatment couch or not. The mean spatial displacement for intrafractional motion for rigid devices is 1.3 mm compared to 1.9 mm for nonrigid devices. The modified Gill-Thomas-Cosman frame performed best at controlling intrafractional patient motion, with a 95% probability of observing a three-dimensional (3D) vector length of motion (v95) of less than 1.8 mm, but could not be evaluated for interfractional motion. All other rigid and nonrigid immobilization devices had a v95 of more than 3 mm for intrafractional patient motion. Interfractional patient motion was only evaluated for the rigid devices. The mean total interfractional displacement was at least 3.0 mm for these devices while v95 was at least 6.0 mm. PMID- 16372419 TI - Abstracts from the 2nd Annual World Congress on the Insulin Resistance Syndrome. November 18-20, 2004. Los Angeles, California, USA. PMID- 16372421 TI - [Abstracts of the 114th Scientific Meeting of Japan Prosthodontic Society, October 1-2, 2005, Niigata, Japan]. PMID- 16372418 TI - Matching and reconstruction of brachytherapy seeds using the Hungarian algorithm (MARSHAL). AB - Intraoperative dosimetric quality assurance in prostate brachytherapy critically depends on discerning the three-dimensional (3D) locations of implanted seeds. The ability to reconstruct the implanted seeds intraoperatively will allow us to make immediate provisions for dosimetric deviations from the optimal implant plan. A method for seed reconstruction from segmented C-arm fluoroscopy images is proposed. The 3D coordinates of the implanted seeds can be calculated upon resolving the correspondence of seeds in multiple x-ray images. We formalize seed matching as a combinatorial optimization problem, which has salient features: (a) extensively studied solutions by the computer science community; (b) proof for the nonexistence of any polynomial time exact algorithm; and (c) a practical pseudo-polynomial algorithm that mostly runs in O(N3) time using any number of images. We prove that two images are insufficient to correctly match the seeds, while a third image renders the matching problem to be of nonpolynomial complexity. We utilize the special structure of the problem and propose a pseudopolynomial time algorithm. Using three presegmented images, matching and reconstruction of brachytherapy seeds using the Hungarian algorithm achieved complete matching in simulation experiments; and 98.5% in phantom experiments. 3D reconstruction error for correctly matched seeds has a mean of 0.63 mm, and 0.9 mm for incorrectly matched seeds. The maximum seed reconstruction error in each implant was typically around 1.32 mm. Both on synthetic data and in phantom experiments, matching rate and reconstruction error achieved using presegmented images was found to be sufficient for prostate brachytherapy. The algorithm is extendable to deal with arbitrary number of images without any loss in speed or accuracy. The algorithm is sufficiently generic to provide a practical solution to any correspondence problem, across different imaging modalities and features. PMID- 16372422 TI - Inferior oblique myectomy, disinsertion and recession in rhesus monkeys. 1977. PMID- 16372423 TI - The National Health Educator Competencies Update Project: celebrating a milestone and recommending next steps to the profession. PMID- 16372425 TI - [Abstracts of the 33rd Annual Meeting of the Austrian Diabetes Association, Baden, 1-3 December 2005]. PMID- 16372424 TI - NO-dependent mechanism of adrenergic reaction of systemic hemodynamics. AB - Blockade of NO synthesis in narcotized rats potentiated pressor effects of phenylephrine by 55% and increased total peripheral resistance by 153%. Vasodilation caused by enhanced NO secretion modulated pressor shifts evoked by stimulation of alpha(1)-adrenoceptors with phenylephrine. PMID- 16372426 TI - MedNet 2005. Abstracts of the 10th World Congress on the Internet in Medicine, Prague, Czech Republic, December 4-7, 2005. PMID- 16372427 TI - UNAIDS statement on South African trial findings regarding male circumcision and HIV. PMID- 16372428 TI - Access to HIV treatment continues to accelerate in developing countries, but bottlenecks persist, says WHO/UNAIDS report. PMID- 16372429 TI - Conversation with Joseph V. Brady. PMID- 16372430 TI - HIV/AIDS information and services: the situation experienced by adolescents with disabilities in Rwanda and Uganda. AB - PURPOSE: The study had two aims: (2) To explore whether knowledge about HIV and AIDS was similar among adolescents with disabilities compared with their non disabled peers; and (2) To determine factors which may increase vulnerability of disabled adolescents to HIV infection and/or inappropriate access to HIV-related services. METHOD: A qualitative study using focus group discussions and sem structured interviews was conducted with purposefully selected participants in Rwanda and Uganda. The participants included disabled adolescents, non-disabled adolescents, parents, teachers, members of disabled people's organisations and representatives of HIV/AIDS organisations. Interviews explored issues of HIV/AIDS knowledge, access to HIV/AIDS services and perceptions of personal risk. RESULTS: Barriers preventing adequate access to information about HIV and AIDS experienced by adolescents with disabilities depended on the nature and severity of the impairment. For example, parents and health workers were unable to communicate with deaf adolescents using sign language, adolescents with physical impairments were often unable to access community meetings about HIV and print material was not adapted for those with visual impairments. Further, assumptions by health workers and community members that people with disabilities were not sexually active led to the marginalisation of disabled people from HIV services. Adolescents with disability described low self-esteem and issues of self-efficacy affecting control of safer sexual relationships. A high level of targeted abuse, rape and exploitation was reported leading to vulnerability among this population. CONCLUSION: The impact of the HIV epidemic among people with disabilities is a neglected area. This study supports the need to develop strategies in HIV prevention programmes that include people with disabilities. PMID- 16372431 TI - Six-minute walk distance as a measure of functional exercise capacity in multiple sclerosis. AB - PURPOSE: We hypothesised that six-minute walk (6MWT) distance of patients with ambulatory multiple sclerosis (MS)would differ from age-matched healthy control subjects. We also investigated the contribution of demographic, physical and physiological factors to impaired functional capacity in MS. METHOD: Thirty MS patients and 30 healthy subjects participated in this study. Respiratory muscle strength was measured. Pulmonary function test and 6MWT were performed. The Barthel Index (BI) was used to assess activities of daily living, and the Modified Ashworth Scale was used to determine spasticity. Symptomatic fatigue was measured using the Fatigue Severity Scale (FSS). RESULTS: Pulmonary function and respiratory muscle strength of ambulatory MS patients were significantly lower, and baseline heart rate and fatigue perception were significantly higher than were healthy controls (p < 0.05). MS patients reached a significantly higher exercise heart rate, and walked significantly shorter distance than did healthy subjects (p < 0.05). The BI score, baseline heart rate and FSS score together accounted for 81 percent variance in 6MWT distance of MS patients (p < 0.05). CONCLUSION: The shorter distance covered during a 6MWT is determined by the limitations in activities of daily living,resting heart rate and subjective symptomatic fatigue in ambulatory patients with MS. Respiratory muscle weakness, lung function and level of neurological impairment do not contribute to impaired functional exercise capacity in these patients. PMID- 16372432 TI - Disability in the upper extremity and quality of life in hand-arm vibration syndrome. AB - PURPOSE: To investigate whether hand-arm vibration syndrome (HAVS) leads to disability in the upper extremity or deficit in quality of life (QoL) using validated questionnaire tools, and to establish whether these effects are related to the Stockholm Workshop Staging (SWS). METHOD: This was a postal cross sectional questionnaire study with a 50% response rate. Four hundred and forty four males, who had been diagnosed and staged according to the SWS were sent the Disability in the Arm, Shoulder and Hand (DASH) and the SF-36v2 QoL questionnaires. RESULTS: HAVS cases had significantly greater DASH disability scores and reduced QoL physical and mental component scores compared to published normal values. Those HAVS cases with a presumptive diagnosis of Carpal Tunnel Syndrome(CTS) had even higher disability scores. There was a clear, linear relationship between both the DASH disability score and the physical component of the QoL and sensorineural SWS, but not with the vascular SWS. CONCLUSIONS: HAVS has a significant effect on an individual's perceived ability to perform everyday tasks involving the upper extremity, and their quality of life. Physical capability may be further compromised in those individuals who have a presumptive diagnosis of CTS. These findings may have important implications regarding management of the affected worker. PMID- 16372433 TI - Abstracts of the Annual Meeting of the Royal Belgian Society for Ear, Nose and Throat, Head and Neck Surgery, 17-19 November 2005. PMID- 16372434 TI - Abstracts of the 95th Annual Meeting of the Poultry Science Association, July 31 August 3, 2005, Auburn, Alabama, USA. PMID- 16372435 TI - Abstracts of the 2005 International Poultry Scientific Forum, January 24-25, 2005, Atlanta, Georgia, USA. PMID- 16372436 TI - United States v. Jacobson. PMID- 16372437 TI - Adams v. State. PMID- 16372438 TI - 66 Federal Credit Union v. Tucker. PMID- 16372439 TI - [Grigorii Viktorovich Gershuni (on his 100th birthday)]. PMID- 16372440 TI - Using the Internet for clinical instruction. PMID- 16372441 TI - The free clinic as a service learning opportunity. PMID- 16372442 TI - Answering the call for abstracts: spontaneous action or deliberate choice? PMID- 16372443 TI - Rating students' technology generated clinical decision making scores. PMID- 16372444 TI - Levels of prevention: restructuring a curriculum to meet future healthcare needs. PMID- 16372445 TI - A decade of distance education: RN to BSN. AB - TELETECHNET represents a major commitment on the part of the university. The university's strategic plan and resource allocations indicate that this educational strategy is a growing enterprise. Although challenges exist in implementing a nursing education program by television, the TELETECHNET program provides opportunities for the place-bound student to achieve advanced education. PMID- 16372446 TI - Lessons from Hurricane Floyd. PMID- 16372447 TI - An icebreaker to introduce nursing theory. PMID- 16372448 TI - Thinking critically about your assignments. PMID- 16372449 TI - Organizational strategies for clinical teaching. PMID- 16372450 TI - A comparison of NLNAC and CCNE accreditation. AB - Baccalaureate and graduate nursing education programs now have a choice of professional accrediting bodies and must make a decision about which best suits the needs of their individual programs. The authors describe the accreditation process and procedures of the National League for Nursing Accrediting Commission and the Commission on Collegiate Nursing Education. The purpose, philosophy, standards, accreditation process, fees, review cycle, and length of accreditation are compared. PMID- 16372451 TI - Dealing with the unsafe student in clinical practice. AB - How do social justice issues affect the clinical evaluation of students? An emotionally charged issue, clinical evaluation can become a mine field when it becomes entangled with issues of social justice. The authors discuss the issues of justice and fairness in relation to situations that arise when a student repeatedly fails to meet minimum expectations for clinical performance. They discuss policies developed to deal with such situations and describe the application of these policies in a case study. PMID- 16372452 TI - Designing and facilitating class discussion in an Internet class. AB - Designing a replacement for face-to-face classroom discussion is a major consideration in the development of an Internet course, whether the course is completely new or a redesign of an existing course. The experiences of four faculty members who were involved in developing courses for a baccalaureate completion program for RNs taught entirely on the Internet will be used to illustrate key issues in designing andfacilitating on-line discussion. PMID- 16372453 TI - Legal issues related to dismissing students for clinical deficiencies. AB - Clinical faculty have a legal, ethical, and professional obligation to evaluate students' clinical performance to ensure safe practitioners. However, many clinical faculty members fear litigation and hesitate to give an unsatisfactory grade to students who do not meet established standards. If the decision to assign a failing clinical grade was "careful and deliberate" and the student was afforded due process, the principle of judicial deference emerges to protect the clinical instructor and the educational institution. PMID- 16372454 TI - Medication administration: a student-centered interactive approach. PMID- 16372455 TI - Developing critical thinking with active learning strategies. AB - The development of students' critical thinking abilities is one of the greatest challenges facing contemporary nurse educators, especially those teaching graduate nursing students. Active learning techniques are suggested to promote critical thinking development. The authors describe how active learning strategies nurtured critical thinking processes in a group of adult nurse practitioner students. PMID- 16372456 TI - Teaching students a pattern of reversals eases the care plan process. PMID- 16372457 TI - Roll the camera: artistic production in the classroom. PMID- 16372458 TI - Oklahoma Nurses Association 2005 House of Delegates resolution. Dangers of leaving children unattended in and around motor vehicles. AB - There is an escalation in the number of fatalities and injuries to children left unattended in and around motor vehicles. Since 1996, more than 225 children, most of them age 3 and under, have died of heat stroke after being trapped inside a parked car. In Oklahoma, SAFE KIDS Coalition has worked to educate parents and the public about leaving children in cars especially when temperatures reach 70 degrees or higher. Currently 12 states have laws that prohibit leaving children unattended in automobiles, but those states do not include Oklahoma. In states where no specific law exists, state and local prosecutors have the discretion to criminally charge adults under existing child endangerment laws. At the 2005 ANA House of Delegates a resolution to adopted so that the American Nurses Association would work with other entities to bring this issue to the public's attention to reduce and prevent future injuries and harm to children. PMID- 16372459 TI - Oklahoma Nurses Association 2005 House of Delegates resolution. Intimate partner violence assessment, intervention and documentation. AB - Intimate Partner Violence (IPV), physical and/or sexual assault of a spouse or intimate partner, has been highly associated with severe health problems such as chronic pain, recurring central nervous system symptoms, injuries, chronic disabilities, gastrointestinal disorders, sexually transmitted diseases, gynecological disorders, unintended pregnancies, depression, substance abuse, and post traumatic stress disorder. Statistically, 40-50% of women in IPV experience both physical and sexual assault. IPV is also associated with 3-13% of pregnancies, posing a threat to health and risk of death for the mother or fetus or both from trauma. Further, 40-60% of female murders in North America are committed by intimate partners. Oklahoma ranks 13th in the nation for women murdered by men in single victim, single offender incidents. IPV associated mortality also includes suicide of women with chronic depression related to violence. Women involved with IPV utilize health care providers and the health care system 3 times more frequently than non-battered women' and generate 92% more health plan costs per year as compared to non-battered women. There is sufficient research concurrence to demonstrate a significant number of women in Oklahoma, the nation and world have severe short and long term physical and mental health effects from IPV. Some victims of IPV may turn to health care professionals for help while other victims are fearful of asking for such assistance. Therefore, it is critical that nurses, as the largest group of health care providers and often the first to come in contact with a victim of IPV, do not miss an opportunity for accurate assessment, appropriate intervention and documentation of IPV in order to provide for a safe, timely and healthy resolution. PMID- 16372460 TI - Oklahoma Nurses Association 2005 House of Delegates resolution. The Nightingale tribute. AB - Nursing is a calling, a way of life. Nursing is a service profession that can not be lived in isolation. Nurses rely on each other for the synergistic effect of teamwork in our efforts of care giving. It is appropriate that we honor our colleagues not only during their career, but also at the end of life's journey. The Nightingale Tribute is a way of honoring our colleagues at the end of life's journey. The Tribute has been fully developed and piloted by the Kansas State Nurses Association. A brochure describing The Nightingale Tribute for use by funeral homes and those presenting the Tribute is available free online at http://www.nursingworld.org/snas/ks/index.htm The purpose of bringing this action report to the ONA HOD is to encourage the use of The Nightingale Tribute in Oklahoma. PMID- 16372461 TI - What is the "taxpayers bill of rights" (TABOR)? PMID- 16372462 TI - Hurricane katrina response: Camp Gruber. PMID- 16372463 TI - Overview of the health and medical response to hurricanes Katrina and Rita. PMID- 16372464 TI - Road trip to Camp Gruber: a nursing experience. PMID- 16372465 TI - Transforming the bitter to better after the storm. PMID- 16372466 TI - Healthy environments for nurses, patients, and the communities where we live. PMID- 16372467 TI - The Clinical Nurse Leader (CNL): staying at the bedside; filling in the gap; and impacting patient and staff outcomes. PMID- 16372468 TI - Nurse manager response to the clinical nurse leader initiative. PMID- 16372469 TI - The DNP: a practicing NP's perspective. PMID- 16372470 TI - Nursing and adult asthma--a matter of life and breath. PMID- 16372471 TI - KICCS: a successful strategy to promote smoking cessation in women during and post pregnancy. PMID- 16372472 TI - Collagen structure in skin from fibromyalgia patients. AB - The distribution and amount of collagen in skin from a non-tender-point area from fibromyalgia patients was assessed by quantitative analysis of amino acids and by electron and light microscopy. Skin biopsies were obtained from the front of the thigh of 27 females who fulfilled the American College of Rheumatology criteria of fibromyalgia and from eight control subjects who were matched for gender, age and physical activity. Amino acids were determined by high-performance liquid chromatography. Electron and light microscopic investigations were carried out to examine tissue structure. Among the collagen-related amino acids, the mean number of hydroxyproline residues per 1,000 residues was 52.5 and 63.4 in fibromyalgia patients and control subjects, respectively (p = 0.050); proline residues were 81.7 and 110.0 (p = 0.006); and hydroxylysine residues were 14.7 and 10.1 (p = 0.002). The total amount of skin protein in proportion to dry tissue weight was 83.4% and 72.6% in fibromyalgia and controls, respectively (p = 0.037). The overall microscopic picture was normal. The lamellar structure of the perineurium and a deficiency in collagen packing in the endoneurium was observed more frequently and to a larger extent in fibromyalgia patients than in controls. In conclusion, there are some differences between the amino acid composition of skin proteins in fibromyalgia patients compared with controls. The amount of collagen may be lower in skin from fibromyalgia patients, and collagen packing in the endoneurium may be less dense. PMID- 16372473 TI - Healing effect of a spray containing Rhealba oat colloidal extract in an in vitro reconstitution model of skin. AB - By using a reconstituted skin model, we aimed to evaluate the efficacy of a spray containing Rhealba oat extract on the rapidity of healing and the extension of the newly formed epithelium. A dermal equivalent was first made in a petri dish by combining skin fibroblasts with collagen type I. Then a punch biopsy as a source of epidermal cells was implanted on this dermal equivalent, where a multilayered epidermis developed. The spray containing Rhealba oat extract was added to the culture medium to evaluate epidermal growth by immunohistochemical analysis of mitotic activity (5-bromo-2'-deoxyuridine [BrDu] incorporation). The extension of the neoepithelium in comparison with untreated reconstituted skin over 22 days was evaluated histologically. On day 12, 16% of positive BrDu basal cells was detected after spray treatment in comparison with 4.2% positive cells in untreated reconstituted skin (p < 0.05). During epidermal differentiation between days 12 and 22, we observed a significant increase in the number of cellular epithelial layers after 16 and 18 days of spray treatment. Moreover the extension of re-epithelialization was also significantly increased after spray treatment on days 16 and 18. In conclusion, our results demonstrate a positive modulation of re-epithelialization on a newly formed epithelium by a spray containing Rhealba oat extract displaying a healing effect. PMID- 16372474 TI - Comparative study of the anti-aging effect of retinaldehyde alone or associated with pretocopheryl in a surviving human skin model submitted to ultraviolet A and B irradiation. AB - In the past few years, the cellular effects of ultraviolet (UV) irradiation induced on skin have become increasingly recognized. Indeed, it is now well known that UV irradiation induces structural and cellular changes in all the compartments of skin tissue. Our aim was to study the anti-aging efficacy of a cosmetic cream containing 0.05% retinaldehyde associated with an antioxidant such as pretocopheryl in comparison with a cream containing only 0.05% retinaldehyde. For this purpose, an ex vivo technique using human skin was used to approximate in vivo metabolic conditions. In this model, human skin was maintained alive by organ culture for 14 days and skin aging was simulated with UV irradiation. Creams were applied to the surface of the epidermis and were compared with nontreated skin. After 14 days, free radical modulation was analyzed by hydroperoxide dosage. Epidermal (laminin) and dermal changes (elastic fibers and collagen) were studied by a histological method. Moreover, to examine collagen synthesis, tritiated proline was added to the culture medium and its incorporation in the newly synthesized collagen was evaluated by Webster's method. The formula containing 0.05% retinaldehyde and pretocopheryl significantly decreased UV-generated free radicals. Repair of laminin, elastic fiber and collagen network was significant and the results were better than those obtained with retinaldehyde alone. An increase of collagen synthesis was also shown with the two creams. PMID- 16372475 TI - Results of the clinical examination of an ointment with marigold (Calendula officinalis) extract in the treatment of venous leg ulcers. AB - The aim of this study was to determine the therapeutic efficacy of marigold (Calendula officinalis) extract on the epithelialization of lower leg venous ulcers. The experiment was carried out in 34 patients with venous leg ulcers. The patients were divided into two groups. In the first (experimental) group, patients were treated with an ointment containing marigold extract, which was prepared in an apparatus devised by Soxleth and was incorporated into a neutral base. Twenty-one patients with 33 venous ulcers were treated. Therapy was applied twice a day for 3 weeks. The second group was a control group that consisted of 13 patients with 22 venous ulcers. In the control group, saline solution dressings were applied to ulcers for 3 weeks. In the experimental group the total surface of all the ulcers at the beginning of the therapy was 67,544 mm2. After the third week the total surface of all the ulcers was 39,373 mm2 (a decrease of 41.71%). In seven patients, complete epithelialization was achieved. In the control group the total surface of all the ulcers at the beginning of the therapy was 69,722 mm2. After the third week the total surface of all the ulcers was 58,743 mm2 (a decrease of 14.52%). In four patients, complete epithelialization was achieved. There was a statistically significant acceleration of wound healing in the experimental group (p < 0.05). The results obtained are preliminary, but they suggest the positive effects of the ointment with marigold extract on venous ulcer epithelialization. PMID- 16372476 TI - Oral administration of cyclosporin A in patients with severe alopecia areata. AB - Alopecia areata is a chronic, nonscarring hair loss condition with an unpredictable course that may cause emotional stress in affected patients. Regarding its pathogenesis, the most accepted theory is that alopecia areata is a T-cell-mediated autoimmune condition that is most likely to occur in genetically predisposed individuals. Cyclosporin A is an immunosuppressive agent that has provided new approaches in the treatment of autoimmune diseases. Hypertrichosis, one of the common side effects of orally administered cyclosporin A, encouraged a number of investigators to use the drug in the treatment of alopecia areata, but the reports on this subject have been controversial. We present a small series of patients with severe alopecia areata treated systemically with cyclosporin A at a dose of 3-5 mg/kg for 6 months as well as their 3-month follow-up after cessation of the drug. PMID- 16372477 TI - Treatment of an extensive superficial basal cell carcinoma of the face with imiquimod 5% cream. AB - The efficacy and safety of imiquimod, an immune-response modifier approved for the treatment of anogenital warts that has antiviral and antitumor activity, in the management of an extensive superficial basal cell carcinoma (sBCC) of the face as an alternative to surgical treatment was evaluated in a 75-year-old male with a 4-year history of a progressively enlarging lesion located on the right temporal region. Imiquimod 5% cream was applied daily until clinical resolution. Histopathological confirmation of clinical diagnosis and of tumor clearance were performed before starting treatment and at the end of treatment, respectively. Moreover, monthly post-treatment follow-up visits were planned. At physical examination, an ovalar, erythematous and slightly infiltrated plaque of 5 x 4 cm in size (approximately 20 cm2), partly eroded and crusted, with a sharp, raised, pearly edge, was evident on the right temporal region of the patient. Histopathological examination of a biopsy specimen showed the typical features of sBCC. Imiquimod 5% cream applied daily for 5 months produced complete clinical and histological clearance. No adverse events but considerable irritation were reported during treatment and no relapses were clinically observed at the 6-month follow-up visit. Our findings confirm current reports from the literature showing imiquimod 5% cream to be an effective treatment for sBCC that is especially valuable in avoiding disfigurement in cases of single large lesions located on the face or in those patients who may not be surgical candidates. PMID- 16372478 TI - Angiopoietin-1 reduces iopromide-induced endothelial cell apoptosis through activation of phosphatidylinositol 3'-kinase/p70 S6 kinase. AB - Radiocontrast media can induce vascular endothelial cell apoptosis. Apoptotic damage to the vascular endothelium is an important mechanism in vascular disease. Several growth factors with anti-apoptotic effects may help protect the vascular endothelium from apoptosis. The present study evaluated whether the radiocontrast agent iopromide induces apoptosis in human umbilical vein endothelial cells and also whether angiopoietin-1 (Ang1) protects against iopromide-induced apoptosis through the p70 S6 kinase-dependent signaling pathway. Iopromide induced apoptosis in vascular endothelial cells in a dose-dependent manner. Ang1 reduced iopromide-induced apoptosis in a dose-dependent manner. Wortmannin and LY294002, phosphatidylinositol 3'-kinase inhibitors, decreased the Ang1-induced anti apoptotic effect. Ang1 mediates the activation of mTOR/ribosomal protein p70 S6 kinase through phosphatidylinositol-3' kinase. Wortmannin and rapamycin, an inhibitor of mTOR, suppressed Ang1-induced p70 S6 kinase phosphorylation and partially inhibited the Ang1-induced anti-apoptotic effect. These results suggest that Ang1 may protect vascular endothelial cells from iopromide-induced apoptosis through phosphatidylinositol 3'-kinase and mTOR/S6 kinase. Pretreatment with Ang1 could help maintain normal vascular endothelial cell integrity before and during systemic radiocontrast administration. PMID- 16372479 TI - Pericyte biology and diseases. AB - Microvessels are composed of two types of cells: endothelial cells and pericytes. Pericytes are elongated cells of mesodermal origin that partially surround the endothelial cells of small vessels. As pericytes contain contractile muscle filaments on their endothelial cell side, they have long been regarded as just microvascular counterparts of smooth muscle cells, thus being implicated in the regulation of capillary tone. However recent understanding of pericyte biology suggests that pericytes play an important role in the maintenance of microvascular homeostasis. Indeed, loss or dysfunction of pericytes has been considered to play an active part in the pathogenesis of various types of disorders. In this study, we review the biology of pericytes and the pathological role of pericyte loss or dysfunction in various devastating disorders such as diabetic retinopathy, atherosclerosis and tumor angiogenesis PMID- 16372480 TI - Renoprotective effects of azelnidipine, a dihydropyridine-based calcium antagonist in advanced glycation end product (AGE)-injected rats. AB - Diabetic nephropathy is a leading cause of end-stage renal disease in industrialized countries. Although the molecular mechanisms for the development and progression of diabetic nephropathy are not fully understood, the formation and accumulation of advanced glycation end products (AGEs) have been considered to play a major role in the pathogenesis of diabetic nephropathy. Hypertension is also an independent risk factor for the progression of diabetic nephropathy. However, functional cross-talk between AGEs and blood pressure and their involvement in diabetic nephropathy remain to be elucidated. In this study, we examined the effects of oral administration of azelnidipine, a commercially available dihydropyridine-based calcium antagonist, on renal injury in AGE treated rats. Administration of azelnidipine inhibited the increase of systolic and diastolic blood pressure levels and urinary N-acetyl-beta-D-glucosaminidase activity in exogenously AGE-injected rats. Furthermore, azelnidipine treatment also prevented glomerulosclerosis in AGE-treated rats. These results indicate that renal damage in AGE-injected rats could be mediated, at least in part, by the elevation of blood pressure. Our present study suggests that azelnidipine would represent a valuable drug for the treatment of diabetic nephropathy by blocking the deleterious effects of AGEs. PMID- 16372481 TI - The effects of alpha-lipoic acid on nitric oxide synthetase dispersion in penile function in streptozotocin-induced diabetic rats. AB - Diabetes-induced erectile dysfunction is one of the most prevalent complications of diabetes in males. alpha-Lipoic acid (ALA) and its reduced form, dihydrolipoic acid, are powerful antioxidants. Data strongly suggest that, because of its antioxidant properties, ALA is particularly suited to the prevention and/or treatment of diabetic complications that arise from overproduction of reactive oxygen and nitrogen. The aim of this study was to investigate the localization of nitric oxide synthetase (NOS) in normal and diabetic rat cavernous smooth muscles and to examine the effects of ALA on them. Rats were divided into four groups: control, diabetic, diabetic plus ALA, and ALA only. Penile tissues were taken 15 days after drug application and examined histochemically and immunohistochemically. Comparison of the control and diabetic groups revealed that the axons of nerve cells were not identified with Masson trichrome in the diabetic group, whereas in the control group NOS localization and immunostaining (endothelial NOS [eNOS]) were normal. Diabetic rats administered ALA showed improvement in Masson trichrome, nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) and eNOS localization compared with untreated diabetic rats. Although there was no difference between the control group and the group administered ALA only, we observed an increase in NADPH-d and eNOS. In erection, eNOS and neuronal NOS (nNOS) may have a significant role. In pathologic conditions, erectile dysfunction may occur as a result of an increase in inducible macrophage-type NOS (iNOS). ALA plays an important role in treatment of erectile dysfunction by decreasing iNOS and increasing other isoforms of NOS. PMID- 16372482 TI - Diffusion tensor imaging: possible implications for radiotherapy treatment planning of patients with high-grade glioma. AB - AIMS: Radiotherapy treatment planning for high-grade gliomas (HGG) is hampered by the inability to image peri-tumoural white-matter infiltration. Diffusion tensor imaging (DTI) is an imaging technique that seems to show white-matter abnormalities resulting from tumour infiltration that cannot be visualised by conventional computed tomography or magnetic resonance imaging (MRI). We propose a new term, the image-based high-risk volume (IHV) for such abnormalities, which are distinct from the gross-tumour volume (GTV). For IHV based on DTI, we use the term IHVDTI. This study assesses the value of DTI for the individualisation of radiotherapy treatment planning for patients with HGG. METHODS: Seven patients with biopsy-proven HGG were included in a theoretical planning exercise, comparing standard planning techniques with individualised plans based on DTI. Standard plans were generated using a 2.5 cm clinical target volume (CTV) margin added to the GTV. For DTI-based plans, the CTV was generated by adding a 1 cm margin to the IHVDTI. Estimates of normal tissue complication probability (NTCP) were calculated and used to estimate the level of dose escalation that could be achieved using the DTI-based plans. RESULTS: The use of DTI resulted in non uniform margins being added to the GTV to encompass areas at high risk of tumour involvement, but, in six out of seven cases, the IHVDTI was encapsulated by the standard CTV margin. In all cases, DTI could be used to reduce the size of the planning-target volume (PTV) (mean 35%, range 18-46%), resulting in escalated doses (mean 67 Gy, range 64-74 Gy), with NTCP levels that matched the conventional treatment plans. CONCLUSION: DTI can be used to individualise radiotherapy target volumes, and reduction in the CTV permits modest dose escalation without an increase in NTCP. DTI may also be helpful in stratifying patients according to the degree of white-matter infiltration. PMID- 16372483 TI - Thoracic radiotherapy for limited-stage small-cell lung cancer: controversies and future developments. AB - Thoracic radiotherapy has an established role in the management of limited disease small-cell lung cancer (LD SCLC). However, essential questions relating to the optimisation of thoracic radiotherapy remain unanswered, including volume of irradiation, optimal total dose, fractionation, timing and sequencing of radiation. This review highlights the need for well-designed multi-national trials aimed at the optimisation and standardisation of radiotherapy for LD SCLC. PMID- 16372484 TI - The use of lung dose-volume histograms in predicting post-radiation pneumonitis after non-conventionally fractionated radiotherapy for thoracic carcinoma. AB - AIMS: To assess the use of lung dose-volume histogram (DVH) parameters (specifically V20Gy) in the prediction of radiation pneumonitis for non conventional fraction sizes used in the treatment of lung cancer. MATERIALS AND METHODS: Patients requiring computed tomography planning for thoracic radiotherapy between January 1999 and January 2002 were identified. The patients receiving radical or high-dose palliative radiotherapy had DVH produced routinely during planning. These were retrospectively reviewed and the case notes accessed for additional pre-treatment parameters, demographics and evidence of radiation pneumonitis. The severity of the pneumonitis was then scored using Radiation Therapy Oncology Group criteria. Data were analysed using the SPSS computer program. RESULTS: One hundred and sixty consecutive patients were reviewed. Ninety patients received hypofractionated treatment (fraction size > 2.5 Gy) and 57 continuous hyperfractionated accelerated radiation therapy (CHART) (fraction size 1.5 Gy). Lung V20Gy values ranged from 3% to 53%, with a median value of 24%. Only six patients reported grade 2, and 16 patients grade 3 pneumonitis. Two patients developed fatal, grade 5 pneumonitis. No correlation between pneumonitis score and V20Gy or other possible predictive factors was found. CONCLUSION: The 15% grade 2-5 pneumonitis rate we document is at the lower end of the spectrum reported in other studies. This suggests that using published data on limiting V20Gy values to reduce the risk of radiation pneumonitis can be extrapolated to planning treatment with non-conventionally fractionated radiotherapy. PMID- 16372485 TI - An audit of clinical resources available for the care of head and neck cancer patients in England. AB - AIMS: Thirty-four cancer networks are now responsible for the delivery of head and neck cancer services in England. A survey of all cancer networks was carried out to define the services available for the diagnosis, treatment and support of patients with head and neck cancer. MATERIALS AND METHODS: Fifty-two cancer units were identified in England from the Cancer Services Collaborative 'Improvement Partnership', and approached through a variety of methods. One questionnaire was circulated to all 34 network leads to establish the services and resources in each network. A second questionnaire was circulated to all known head and neck teams in the UK. RESULTS: Multidisciplinary team (MDT) working was evident in all centres, although frequently the MDT was not fully staffed, especially in areas of national staff shortages, such as in pathology, radiology, dietetics and speech therapy. Rapid referral pathways were present for initial assessment of patients, but clinically significant delays were identified in complex investigations, access to intensive care beds and oncological care, especially radiotherapy. CONCLUSION: Major changes in service configuration are likely to be required if the current national targets of time to diagnosis and treatment are to be met. PMID- 16372486 TI - Parotid-sparing radiotherapy: does it really reduce xerostomia? AB - AIMS: Parotid-sparing radiotherapy (PSRT) was introduced for patients with selected head and neck cancer requiring bilateral upper-neck irradiation at our centre in 2000. The aim of this study was to compare the subjective degree of xerostomia in patients treated with PSRT between January 2000 and June 2003 with patients treated using conventional techniques (radiotherapy) over the same period. MATERIALS AND METHODS: Eligible patients were required to have completed treatment 6 months previously and be recurrence-free at the time of interview. PSRT was defined as conformal radiotherapy, in which the mean dose to at least one parotid gland was 33 Gy or less, as determined by the dose-volume histogram. Patients receiving radiotherapy were treated with standard parallel-opposed fields, such that both parotids received a minimum of 40 Gy. Xerostomia was assessed using a validated questionnaire containing six questions with a rating between 0 and 10. Lower scores indicated less difficulty with xerostomia. RESULTS: Thirty-eight eligible patients treated with PSRT were identified: 25 with oropharyngeal cancer and 13 with nasopharyngeal cancer (NPC). The mean overall questionnaire score (Q1-5) for this group was 4.20 (standard error = 0.33). Forty-four patients (24 oropharyngeal, 21 NPC) treated with radiotherapy over the same period were eligible. The mean overall questionnaire score (Q1-5) for this group was 5.86 (standard error = 0.35). The difference in mean overall scores between the two groups of patients was statistically significant (P < 0.001), as were the scores for four of the six individual questions. CONCLUSION: These results suggest that PSRT offers improved long-term xerostomia-related quality of life compared with conventional radiotherapy. PMID- 16372487 TI - A randomised study of whole-breast vs tumour-bed irradiation after local excision and axillary dissection for early breast cancer. AB - AIMS: Whole-breast radiotherapy (WBRT) after conservative surgery for early breast cancer is a routine standard of care. Despite this, a number of uncertainties in management still exist. Over recent years, a number of new technologies have allowed the development of partial-breast irradiation, with the intention of improving the risk-benefit relationship of routine breast radiotherapy. We report the results of a trial comparing partial- with WBRT, with prolonged follow-up. MATERIALS AND METHODS: Between 1986 and 1990, 174 women were randomised to receive conventional whole-breast radiotherapy (WBRT) (40 Gy in 15 fractions), with a tumour-bed boost or partial-breast irradiation by a variety of techniques. Recruitment was problematic, and the trial closed prematurely well before meeting its recruitment target. RESULTS: A trend was observed towards higher local recurrence and a higher locoregional recurrence rate after irradiation of the tumour bed alone. Distant recurrence and survival were the same. CONCLUSIONS: Conclusions are limited in view of the failure to complete accrual of the target of 400 participants, and in the context of the techniques of partial-breast radiotherapy used during this study, which would not compare with those in current use. Tumour-bed irradiation alone cannot currently be recommended as routine treatment outside the context of clinical trial. PMID- 16372488 TI - Postmastectomy radiotherapy reduces locoregional recurrence in elderly women with high-risk breast cancer. AB - AIMS: Clinical trials of adjuvant radiotherapy after mastectomy have largely excluded women aged 70 years or over, even though they comprise 30% of the breast cancer population. This study examined outcomes in elderly women with high-risk breast cancer treated with or without postmastectomy radiotherapy (PMRT). MATERIALS AND METHODS: Data were analysed for 233 women aged 70 years or over with high-risk breast cancer (tumours > 5 cm or > or = 4 positive axillary nodes) treated with mastectomy and referred to the British Columbia Cancer Agency from 1989 to 1997. Tumour and treatment characteristics were compared between two cohorts: women treated with PMRT (n = 147) vs women treated without PMRT (n = 86). Univariate and multivariate analyses of 10-year Kaplan-Meier locoregional recurrence (LRR), distant recurrence, breast cancer-specific survival and overall survival were carried out. RESULTS: Median follow-up time was 5.5 years. The distribution of tumour sizes was similar in the two groups. Compared with women treated without PMRT, higher proportions of women who underwent PMRT had four or more positive nodes (83% vs 67%, P = 0.01) and positive surgical margins (14% vs 4%, P = 0.02). Systemic therapy, used in 94% of women, was comparable in the two cohorts (P = 0.63). Elderly women treated with PMRT had significantly lower LRR compared with women treated without PMRT (16% vs 28%, P = 0.03). No differences in distant recurrence, breast cancer-specific survival or overall survival were observed in the two treatment groups (all P > 0.05). On multivariate analysis, the omission of PMRT and the presence of high-grade histology were significant predictors of LRR, whereas an increasing number of positive nodes was significantly associated with distant recurrence and overall survival. CONCLUSIONS: In women aged 70 years or over with tumours greater than 5 cm or four or more positive nodes, significantly lower LRR was observed in women treated with radiotherapy compared with women treated without radiotherapy. PMRT should be considered in the management of elderly women with these high-risk characteristics. PMID- 16372489 TI - High efficacy of gemcitabine and cisplatin plus trastuzumab in patients with HER2 overexpressing metastatic breast cancer: a phase II study. AB - AIMS: Effective and tolerable regimens are sought specifically in women who have been pre-treated with anthracyclines and taxanes. Gemcitabine and cisplatin plus trastuzumab has shown synergistic activity in vitro, and provides a new mechanism of drug interaction. This multicentre phase II study aimed to evaluate the efficacy and tolerability of gemcitabine and cisplatin plus trastuzumab in previously treated patients with metastatic breast cancer (MBC). MATERIALS AND METHODS: Previously treated patients with human epidermal growth factor receptor 2 (HER2) overexpressing MBC were enrolled in a multicentre phase II study (DAKO Hercep Test 3+). Treatment consisted of gemcitabine (750 mg/m2), cisplatin (30 mg/m2) given on days 1 and 8 every 3 weeks, and trastuzumab (4 mg/kg loading dose, 2 mg/kg weekly). RESULTS: Twenty patients were recruited, all of whom had previously received chemotherapy (12 pre-treated with taxanes, 18 pre-treated with anthracyclines seven pre-treated with taxanes and trastuzumab). A median of six cycles of the study treatment was delivered. There were eight partial responses, for an overall response rate of 40% (95% confidence interval 16.5 63.5%). The clinical benefit rate (complete response plus partial response plus stable disease) was 80% (95% CI 54.2-95.8%). The response rate in patients who had already received a trastuzumab-based regimen for MBC was 57.1% (95% CI 7.7 100%). Median time to progression was 10.2 months, and median overall survival was 18.8 months. Main toxicities were leukopenia (grade 3 in 55% of patients) and thrombocytopenia (grade 3 in 35% and grade 4 in 5% of patients). Non haematological toxicity was rarely severe. CONCLUSIONS: Combination chemotherapy with gemcitabine and cisplatin plus trastuzumab is well tolerated and active in patients with HER2 overexpressing MBC, even after prior exposure to anthracyclines and taxanes. PMID- 16372490 TI - Human-immunodeficiency-virus-negative, human-herpes-virus-8-negative abdominal cavity primary effusion lymphoma. AB - Primary effusion lymphoma (PEL) is an unusual class of non-Hodgkin's lymphoma, which is characterised by lymphomatous effusions in body cavities, but no associated mass lesions. It is usually associated with an immunodeficient state most often with the human immunodeficiency virus (HIV). We describe a case of a man with HIV-negative, human-herpes-virus-8 (HHV8)-negative PEL, with a history of heavy alcohol intake. The abdominal cavity was the only area involved; no solid tumour masses were observed on scanning, and the bone-marrow investigations were normal. The ascites contained numerous pleomorphic lymphoid, lymphoplasmacytoid cells of B-cell origin. The immunophenotyping was moderately positive for CD 38 and 138, and strongly positive for Ki 67. It is postulated that the immunosuppressed state in this patient may have been caused by the long history of heavy alcohol intake. PMID- 16372491 TI - Nutrition support improves patient outcomes, treatment tolerance and admission characteristics in oesophageal cancer. AB - AIMS: Patients with oesophageal cancer undergoing chemoradiation with curative intent are at high risk of malnutrition and its complications, including increased side effects of treatment. We have developed a nutrition pathway (NP), involving the early then periodic nutrition assessment of all patients presenting to the multidisciplinary oesophageal clinic who were planned to receive definitive chemoradiation. MATERIALS AND METHODS: Patients were assessed as at 'low', 'moderate' or 'severe' nutrition risk, and were provided with appropriate nutrition intervention ranging from preventative advice (low risk), oral nutrition support (moderate risk) to enteral feeding (severe risk). Outcomes for 24 patients treated before implementation of the NP were compared with those of 24 patients treated using the NP. RESULTS: Patients managed using the NP experienced less weight loss (mean weight change -4.2 kg +/-6.4 cf. -8.9 kg +/- 5.9, P = 0.03), greater radiotherapy completion rates (92% cf. 50%, P = 0.001), fewer patients had an unplanned hospital admission (46% cf. 75%, P = 0.04), and those that did had a shorter length of stay (3.2 days +/- 5.4 cf. 13.5 days +/- 14.1, P = 0.002). CONCLUSION: Early and regular nutrition assessment/intervention and a multidisciplinary approach to nutrition care results in improved treatment tolerance for patients with oesophageal cancer receiving chemoradiation. PMID- 16372492 TI - Intravenous irinotecan plus oral ciclosporin. AB - We previously reported a phase I study of intravenous irinotecan plus oral ciclosporin, in which dose-limiting diarrhoea was not observed, supporting the hypothesis that pharmacokinetic modulation of irinotecan by ciclosporin may improve its therapeutic index. We now report results of a further 34 patients treated at the recommended dose. A low rate of diarrhoea of grade 3 or above (3%) was again seen, with useful anti-tumour activity. The regimen is to be formally evaluated as part of a future phase III trial. PMID- 16372493 TI - Genetic predisposition to cancer: the consequences of a delayed diagnosis of Gorlin syndrome. AB - This report outlines a case of Gorlin syndrome, the diagnosis of which was delayed for many years, and raises a number of important issues. These are the spectrum of late radiotherapy effects, particularly after treatment for benign disease, and the importance of considering the possibility of the presence of a genetic syndrome predisposing to cancer in all individuals before starting any treatment. As our knowledge of genetic syndromes expands, this will become increasingly important. Finally, if a genetic predisposition to cancer is suspected, consideration should be given to obtaining a blood sample from the affected patient for DNA storage, particularly if their prognosis is limited. Currently, genetic testing can only be instituted in most families by first obtaining DNA from an individual affected by cancer, as most genetic mutations are unique to a family. If all relatives with cancer have died, then, at this time, genetic testing cannot usually be attempted, unless such samples have previously been stored. PMID- 16372494 TI - Gleason scoring varies among pathologists and this affects clinical risk in patients with prostate cancer. AB - AIMS: To investigate whether our practice of specialist review of all diagnostic biopsies was necessary to prevent misgrading of referred prostate cancer patients, and whether this misclassification, if any, would have resulted in misclassification of clinical risk grouping (Seattle Risk Grouping [SRG]) and subsequent treatment strategy and prognosis. MATERIALS AND METHODS: Important prognostic indicators for prostate cancer include the presenting prostate specific antigen (PSA), clinical stage and Gleason sum of the tumour. These three variables are incorporated into the SRG cohorts to establish treatment strategy. Patients with prostate cancer referred for brachytherapy had their prostate biopsies reviewed by a reference pathologist (PD) with a special interest in prostate cancer. We compared the agreement between the scoring of the referring pathologists with that of PD, and evaluated if any differences changed the SRG and therefore the clinical risk and treatment strategy for the patients. RESULTS: In only 52% (43/83) of cases, was there total agreement between the two sets of pathologists. The inter-rater agreement was statistically 'fair' (unweighted kappa statistic 0.27). In 90% (36/40) of cases with disagreement, PD assigned higher Gleason sums. In 40% (16/40) of cases with disagreement, the change in Gleason sum altered the SRG; in one out of 16 cases, the SRG was downgraded from 'intermediate' to 'low' risk disease; in six out of 16 cases, it was upgraded from 'low' to 'intermediate' risk, and, in nine out of 16, from 'intermediate' to 'high' risk. CONCLUSION: Our findings confirm previous reports of only limited correlation between pathologists in reporting Gleason sums. In this study, 19% (16/83) of cases had their grading changed to a level that altered clinical risk, almost always (94%; 15/16) to one that worsened prognosis. This would have significantly affected treatment strategy for these patients, and thus we recommend that all centres ensure accurate Gleason grading by the use of pathologists with special interests in prostate cancer. PMID- 16372495 TI - A method for assessing accurate application of the Partin Tables in the pre therapy evaluation of patients with prostate cancer. AB - AIMS: The treatment of prostate cancer is frequently influenced by the Partin Tables. This predictive model has been internally and externally validated since it was conceived, and has proved to be remarkably reliable and consistent. This paper proposes that, by using the statistical programme for the social sciences (SPSS) and receiver-operator characteristic curves, it is possible to detect institutions that apply this model sub-optimally. MATERIALS AND METHODS: This theory was supported by a PUBMED search using relevant search words. RESULTS: This is a novel technique with the potential to allow retrospective and prospective accrual of results. CONCLUSIONS: A systematic institutional review of how accurately a hospital assesses the clinical stage, Gleason score and PSA has the potential to increase an institution's predictive accuracy when it uses the Partin Tables. The proposed method allows for quantitation of the level of error and comparison of predictive accuracy between institutions. It also may be used as an internal outcome measure to assess improvement in a hospital's investigative procedures over time. PMID- 16372496 TI - Chemotherapy in a patient with small-cell lung cancer undergoing haemodialysis. PMID- 16372497 TI - Intra-arterial chemotherapy in elderly patients with invasive bladder cancer. PMID- 16372498 TI - Use of orfit to reduce the irradiated lung volume in patients receiving radiotherapy after breast-conserving surgery. PMID- 16372499 TI - Neurogenic pulmonary oedema mimicking an anaphylactic reaction to interferon alpha-2b. PMID- 16372500 TI - Renal cell carcinoma presenting as an isolated neck mass. PMID- 16372501 TI - Irinotecan with bolus and infusional 5-flurouracil and folinic acid for patients with advanced or metastatic colorectal cancer previously treated with 5 flurouracil: a possible alternative to single-agent irinotecan in a 'real-life' setting. PMID- 16372502 TI - Administration of cisplatin and etoposide during chronic ambulatory peritoneal dialysis. PMID- 16372503 TI - Non-oncology specialists may incorrectly attribute symptoms to previous radiotherapy. PMID- 16372504 TI - Multiple malignancies and immunological diseases after radiotherapy: a new tumour suppressor gene disorder? PMID- 16372505 TI - Role of risk factors for erectile dysfunction in patients undergoing transurethral resection of the prostate: early impact on sexual function. AB - INTRODUCTION: Transurethral resection of the prostate (TURP) has been long debated as a possible cause of erectile dysfunction (ED). We investigated the role of common risk factors for ED in patients aged 60 to 70 undergoing TURP Factors related to the treatment were also considered. MATERIALS AND METHODS: Ninety patients underwent TURP for benign prostate hyperplasia (BPH) from June 2002 to February 2003. Fourty-two of them, sexually active aged 60 to 70, were administered preoperatively and 3-month postoperatively the International Prostate Symptom Score (IPSS) and International Index of Erectile Function (IIEF 5) questionnaires. A complete assessment of risk factors for ED was performed in the preoperative setting (treated hypertension, diabetes, cigarette smoking, hypercolesterolemy, cardiovascular disease). IIEF score was related to age, comorbidities, operative time, resected tissue weight, retrograde ejaculation, IPSS score. RESULTS: Nine (21.4%) patients reported worsened IIEF-5 score after TURP, and 33 (78.6%) unchanged/improved score. Cardiovascular disease was present in 56% of patients with worsened IIEF-5 score and in 12% of patients with improved/unchanged IIEF-5 score; it was the only factor that correlated significantly in the regression model. CONCLUSION: In general, most patients report a stable sexual function after TURP. Patients with known cardiovascular disease undergoing TURP had an increased risk of sexual impairment after this procedure. PMID- 16372506 TI - Our experience in endoscopic treatment of vesico-ureteral reflux in children. AB - AIM OF THE STUDY: To define the guidelines of endoscopic treatment of vesico ureteral reflux (VUR) in children in relation to grade of VUR, of the location of the ureteral orifice, of the stage of reflux nephropathy and of the association with other urinary tract malformations. MATERIALS AND METHODS: 498 children with vesico-ureteral reflux (VUR) were observed in 702 ureters. All the patients were submitted to a complete urological evaluation and to endoscopic treatment (ET) of the VUR with the implant of a stable polyacrylamid gel (DAM+). RESULTS: The reflux was degree I in 53 ureters, II in 174, III in 301, IV in 165 and V in 9 ureters. VUR was primary in 149 children, secondary to a neurogenic dysfunction of the bladder in 271 patients, complex in the remaining cases. An overall 90.5% success rate of endoscopic treatment with "DAM+" implantation was observed. Success of VUR endoscopic treatment was obtained in 80% of cases with- orifice lateralisation, in 60% of cases with high and low intravesical orifice ectopy, and in 10% of cases with extravesical orifice. In 151 ureters (21.5%) endoscopical treatment of VUR had to be repeated, while a third procedure was necessary in 42 ureters (5.9% of cases). In total 895 endoscopic procedures were performed. Open surgical ureteral reimplantation was performed in the cases with persisting VUR after the third attempt of endoscopic procedure. CONCLUSION: These results of the endoscopic treatment of VUR in children confirm the high efficacy and safety of this method. In any case, before choosing the method of VUR correction, it is necessary to inform the parents of the child about the potential rate of success of the endoscopic treatment in function of the peculiarity of the individual patient. PMID- 16372507 TI - Detection rate of ultrasound vs CT scan in clinical staging accuracy of renal tumors pT1NxMx. AB - OBJECTIVE: Renal ultrasound is very important in the diagnosis of renal tumors. MATERIAL AND METHODS: From January 2000 to Jannuary 2005 we retrospectively examined the records of 116 patients, 37 women and 79 men (mean age 54 years, range 22-77), who underwent radical nephrectomy for kidney cancer in clinical stage CT1N0M0. 2.5 cm was the tumor dimension limit between the nephron sparing surgical technique and radical nephrectomy. We subdivided the sample into 2 groups, the first of 45 patients with tumor lesions smaller than 2.5 cm and the second with tumor lesions between 2.5 cm and 7 cm in diameter all patients underwent preoperative staging including ultrasound scan (ETG) and computer tomography scan (CT). RESULTS: Ultrasound has showed 35% sensitivity and 49% specificity for lesions under 2.5 cm in diameter, and 65% sensitivity and 75% specificity and 80% specificity for lesions under 2.5 and 80% sensitivity and 95% specificity for lesions between 2.5 and 7 cm. CONCLUSIONS: Ultrasound is the first step in the staging of renal cancer before surgery; to assess. 2.5 and 7 in diameter CT scan is the gold standard for solid and cystic lesions. PMID- 16372508 TI - A case report of a "smouldering" uretero-vascular fistula. AB - Uretero-vascular fistulae are rare. As compared to aortic, uretero-iliac fistulae are by far more frequent according to a 1:8 ratio. We present one case of uretero prosthetic fistula in a man operated upon the resection of an aortobisiliac aneurysm and Dacron prosthetic graft; six years later, a blunt trauma to the body was followed by a threatening shock, resistant to medical measures. Two subsequent emergency operations allowed to a difficult diagnosis and a life saving repair of the fistula. Some hypotheses on the pathogenesis of the condition are discussed as well as its clinical problems. PMID- 16372509 TI - A rare case of penile fracture with complete urethral rupture during sexual intercourse. AB - Fracture of the penis during intercourse is a relatively uncommon condition. We report a rare case with laceration of bilateral corpora cavernosa and associated complete urethral rupture. The patient underwent immediate surgical repair of the penile fracture with primary urethroplasty. After 1 year follow-up he presents excellent results with normal sexual function and normal postoperative urethrogram with no voiding problems. PMID- 16372510 TI - Sepsis due to asymptomatic Candida prostatitis. AB - A case of a asymptomatic prostatitis due to Candida Albicans that caused a sepsis is presented. Up today in literature only 3 cases of Candida infections of the prostate gland without general illness were described. In this case the transurethral electro-resection of prostate was the adequate treatment. PMID- 16372511 TI - Cyproterone acetate in the therapy of prostate carcinoma. AB - Prostatic carcinoma is one of the most commonly neoplasm in men, with the strongest incidence around the age of 70 years. In consideration of the high hormonodependence of prostatic neoplasm the reduction of testosterone levels represents the choice of treatment for the patients with metastatic disease, and has an application also in the treatment of patients with a more limited disease, but not elegible to local treatment with a curative aim. The circulating testosterone can be maintained to the lowest levels by the use of drugs that can obtain, with different mechanism of action, a "medical castration". Cyproterone acetate (CPA) is a steroideal antiandrogen which has affinity with progesteron and with glucocorticoidal receptors. It centrally inhibits the release of lutehinizing hormon blocking in this way the secretion of testosterone of testicular origin. Besides, it inhibits the action of the androgens of surrenalic and testicular origin to the cellular level through a competitive direct interaction with the cellular receptors. This review illustrates the main evidences of efficacy and safety of CPA in the treatment of prostatic carcinoma. PMID- 16372512 TI - Trends in prostate cancer epidemiology in the year 2000. AB - In North America and Europe during the last 20 years prostate cancer incidence rates progressively increased, peaked in early 1990s and then declined. Newly diagnosed prostate cancers are being detected at an early stage in men presenting with no symptoms with abnormal prostate specific antigen (PSA) level. The proportion of patients treated by radical prostatectomy has increased and the proportion of those treated with hormonal therapy has decreased. The widespread of PSA testing seems to be the cause of the changes in the prostate cancer incidence rates and of the stage migration towards localized disease, but the effective benefit of prostate cancer screening in terms of reduction of cancer related mortality is still to be demonstrated. Definitive evidence on the effectiveness of prostate cancer screening with PSA should come from the results of large-scale screening trials that are still ongoing. In Italy the total number of newly diagnosed cases has been steadily increasing from the early 1980s to the mid 1990s both in Lombardia and Umbria regions. Survival rates dramatically improved from 37-48% to 74-76%, but no trendfor mortality was observed. PMID- 16372513 TI - The natural history: how has prostate cancer trend modified? AB - The patients' number submitted to prostate fine-needle biopsy has constantly increased in the last years, posing technical and diagnostic problems to the pathologist. The use of immunohistochemical tests can help to solve some questions. In other cases, when only a few atypical glands are present, a very cautious behaviour is the rule. The acronym "ASAP" (atypical small acinar proliferation) has been suggested in such cases. PMID- 16372514 TI - Early diagnosis of prostatic cancer: disease-related survival improvement or extension of observation time? AB - It is well recognized that prostate cancer represents the most common cancer in men in western countries and it is the second leading cause of cancer death. Although prostate cancer detection has improved, we still have to establish to what extent our diagnostic tools are correlated with the clinical prognosis. This task of distinguishing more aggressive disease from relatively benign cases remains a major goal. The findings of ongoing randomised screening studies should aid us in this task, so that we can avoid the treatment of those who do not need it and tailor our treatment strategy more closely to the individual prognosis. PMID- 16372515 TI - Detection and diagnosis of prostate cancer: what's new. AB - OBJECTIVE: Prior the widespread use of PSA screening in asymptomatic men, prostate cancer was historically detected by a simple digital rectal examination. Although the gold standard for prostate cancer still remains prostate biopsy, current researches in the area of detection and diagnosis of prostate carcinoma are focusing on identification of better sampling protocols, biologic markers and imaging strategies in order to detect disease at an earlier stage. We reviewed all the recent literature on the detection of clinically meaningful prostate cancer. METHODS: A systematic review of the literature using Medline up to 2005 was performed. Electronic searches were limited to the English language using the keywords prostate cancer, diagnosis, transrectal ultrasound, prostate biopsy. Unpublished information known by the authors and that were considered of interest to the readers were also included. RESULTS: The prostate biopsy technique has extremely changed from the original Hodge's sextant biopsy protocol. Several authors have already reported high rates of false negative biopsy using sextant protocols. The optimal protocol should, nowadays, include six standard sextant biopsies with additional biopsies weighted more laterally (anterior horn) and medially to the apex. Repeat biopsies should also be based on an extended scheme and should include the transition zone especially in patient with at initial negative biopsy. To increase accuracy of prostatic biopsy and reduce unnecessary prostate biopsy, TRUS, power Doppler imaging (PDI), colour Doppler TRUS (CDUS), and 3-dimensional Doppler (3DD) can be successfully adopted, but their routine use is still controversial. Several types of local anaesthesia are now available and can be safely performed to reduce the pain of multi-sites biopsy protocol. CONCLUSION: Extended biopsy schemes should be performed not only at first biopsy but especially at repeated biopsy for premalignancy lesions. The widespread use of local anesthesia makes the procedure more comfortable. PMID- 16372516 TI - The ability of statins to protect low density lipoprotein from oxidation in hypercholesterolemic patients. AB - OBJECTIVE: It is unclear at the present time whether hydroxy-methylglutaryl coenzyme A reductase inhibitors (HMG-CoA reductase inhibitors; statins) exert a protective effect on low-density lipoproteins (LDL) oxidation in vivo. In addition, it is speculated that pharmacological differences between statins may account for differences in their antioxidative capacities. This is of clinical relevance, because there is strong evidence that oxidized LDL initiates the atherosclerosis process. MATERIAL AND METHODS: In a controlled, randomized, double-blind study we compared the effects of three different statins (simvastatin, pravastatin and atorvastatin) on the ability to protect LDL from oxidation in 70 hypercholesterolemic but otherwise healthy subjects. Statins were administered in doses which were nearly equi-effective in lowering LDL cholesterol. Changes in LDL oxidation were measured using diene conjugation (DIENES) and thiobarbituric acid reactive substances (TBARS) at entry and three months after beginning therapy with the statins. RESULTS: Levels of DIENES, usually generated during the early phases of lipid peroxidation, were significantly reduced by 10.2 +/- 5.5% (mean +/- SEM; p < 0.03), 6.0 +/- 2.0% (p < 0.005) versus baseline in the case of pravastatin and atorvastatin but simvastatin had no significant effect with a mean reduction of 5.5 +/- 6.4% (p > 0.23). Levels of TBARS, reflecting late phases of LDL oxidation, showed no significant changes against baseline (p > 0.34). Pooled data (n = 70) indicated that statins reduce DIENES levels by approximately 9% versus baseline (p < 0.005) but had no significant effect on TBARS levels (p > 0.29) after three months of therapy. CONCLUSION: This study showed that atorvastatin and pravastatin were capable of protecting LDL from oxidation in vivo in the early treatment phase. Pooled data levels of DIENES were significantly affected by statin therapy over a period of 3 months. No protective effect appeared to be present in the late phases of oxidation evaluated using measurement of TBARS but it should be noted that the clinical impact of such observations are currently discussed controversially in the literature. PMID- 16372517 TI - Acute pancreatitis associated with omeprazole. AB - Since their introduction in the late 1980s, proton pump inhibitors (PPI) have demonstrated gastric acid suppression superior to that of histamine H2-receptor blockers. This class of drugs has improved the treatment of various acid-peptic disorders, including gastroesophageal reflux disease, peptic ulcer disease, and nonsteroidal anti-inflammatory drug-induced gastropathy. PPIs have minimal side effects and few significant drug interactions. They are generally considered safe for long-term treatment. We present a rare side effect, acute pancreatitis, occurring in a patient who was treated with the proton pump inhibitor omeprazole. PMID- 16372518 TI - The pharmacokinetics of zinc from zinc gluconate: a comparison with zinc oxide in healthy men. AB - OBJECTIVE: Zinc supplementation is beneficial in some clinical conditions such as age-related macula degeneration (AMD). It has been suggested that zinc absorption is influenced by the form in which zinc is ingested. Therefore, the pharmacokinetics of zinc gluconate (organic) were compared with those of zinc oxide (inorganic). METHODS: 12 healthy male subjects aged between 21 and 31 years (24 years median) orally received daily doses of 20 mg metal zinc as zinc gluconate and 17.4 mg metal zinc as zinc oxide under randomized crossover conditions for 14 days each with at least 14 days as a washout. Zinc plasma concentrations were measured by means of inductively coupled plasma-atomic emission spectroscopy. RESULTS: C(max) was found 18.3% (10.3 - 26.3%) higher following multiple-dose administration of zinc gluconate as compared to zinc oxide (mean; 0.95% confidence interval of the relative differences between both treatment conditions; p < 0.05). AUC(0-24h) was noted 8.1% (1.9 - 14.3%) higher after zinc was given as zinc gluconate when compared to zinc oxide (p < 0.05) whereas t(max) did not differ between both treatment conditions. CONCLUSIONS: Zinc absorption in humans could be improved by zinc complexation with gluconate. PMID- 16372519 TI - Pemetrexed: mRNA expression of the target genes TS, GARFT and DHFR correlates with the in vitro chemosensitivity of human solid tumors. PMID- 16372520 TI - In vitro investigation on the selectin binding mechanisms in tumor cell metastasis and their inhibition by heparin. PMID- 16372521 TI - Individual variation in factors affecting the steps between dose application and effects of antineoplastic agents. PMID- 16372522 TI - Rational development of oxaliplatin analogues --synthesis and preliminary structure-activity relationships. PMID- 16372523 TI - Current pharmacogenetics in colorectal cancer--do they have an impact on our clinical treatment strategies? PMID- 16372524 TI - Identification of new genes involved in cisplatin resistance and their expression profile in 18 human tumor cell lines. PMID- 16372525 TI - Targeting of immunoliposomes to endothelial cells expressing VCAM: a future strategy in cancer therapy. PMID- 16372526 TI - Interactions of a novel ruthenium-based anticancer drug (KP1019 or FFC14a) with serum proteins--significance for the patient. PMID- 16372527 TI - A new oxygen-enriched solution enables human tumor tissue transport without cell devitalization. PMID- 16372528 TI - Pharmacokinetics (PK) of a liposomal encapsulated fraction containing doxorubicin and of doxorubicin released from the liposomal capsule after intravenous infusion of Caelyx/Doxil. PMID- 16372529 TI - Early results from a phase I study on orally administered tris(8 quinolinolato)gallium(III) (FFC11, KP46) in patients with solid tumors--a CESAR study (Central European Society for Anticancer Drug Research--EWIV). PMID- 16372530 TI - Molecular targets of indirubins. PMID- 16372531 TI - KP1019 (FFC14A) from bench to bedside: preclinical and early clinical development -an overview. PMID- 16372532 TI - Expression profile of copper transporters in sensitive and cisplatin/oxaliplatin resistant tumor cell lines. PMID- 16372533 TI - Proteomic analysis of chemoresistance to 5-fluorouracil in colon cancer in vitro. PMID- 16372534 TI - Flat dose (175 mg/weekly) paclitaxel: pharmacokinetics and clinical implications. PMID- 16372535 TI - Enzastaurin and pemetrexed exert synergistic antitumor activity in thyroid cancer cell lines in vitro. PMID- 16372536 TI - Contrast-enhanced MR imaging in the indirect monitoring of drug release from an interstitial depot. PMID- 16372537 TI - Searching for cellular targets of novel pteridines downstream of cAMP PDE inhibition. PMID- 16372539 TI - The Katrina effort. PMID- 16372540 TI - Disclosing information to parents of minor patients. PMID- 16372538 TI - Anticoagulants: another option. PMID- 16372541 TI - When it comes to morale, you get what you give. PMID- 16372542 TI - "Engage" your patients for a better dental office experience. PMID- 16372543 TI - Blue Cross rate renewal numbers looking good. PMID- 16372544 TI - Post-menopause, periodontal disease and tooth loss. PMID- 16372546 TI - Members want to know...we answer your most-asked questions. PMID- 16372547 TI - Jaw osteonecrosis related to bisphosphonates: a new concern for dentistry. PMID- 16372548 TI - Adverse oral sequelae to bisphosphonate administration. AB - Osteonecrosis of the jaws following the administration of bisphosphonate class drugs, both intravenous and oral, has recently been reported in the literature. A case series of five patients is presented. The clinical presentation, management, and proposed etiology of this adverse reaction are reviewed. Recommendations for identifying patients who may be at risk via medical history, present and past medications, specific drugs involved and recommended precautions involving various dental interventions are also discussed. PMID- 16372549 TI - Deference is dead. PMID- 16372550 TI - Review of the year. Gutsiness, originality and a willingness to fight for justice were behind many of this year's triumphs in nursing. PMID- 16372551 TI - 'Ordinary people can make a difference'. PMID- 16372552 TI - New cases show HIV/AIDS needs a dual approach. PMID- 16372553 TI - Routes to success. AB - New routes into nursing are enticing more people into the profession. PMID- 16372554 TI - Prescription for love. AB - Harlequin Mills & Boon Limited is looking to recruit nurses to write realistic medical romances for one of its highly popular fiction series. PMID- 16372555 TI - Sick and tired. AB - Severe anaemia can blight the lives of people with kidney disease. While effective treatment is available, its provision is patchy across the country. It has been found that: In people with renal anaemia, haemoglobin can fall to 4g/dl. Injections of erythropoietin (EPO) can increase haemoglobin levels. Patients are often unaware of what their haemoglobin levels are and should be. Early treatment with EPO can slow progression of renal disease. PMID- 16372556 TI - The gift of Christmas presence. PMID- 16372557 TI - Patient safety: improving incident reporting. AB - The aim of this article, the fourth of seven in this series, is to discuss the reporting of patient safety incidents, including near miss events. This article is based on step four of the National Patient Safety Agency's (2004) Seven Steps to Patient Safety. Barriers to reporting are discussed and the associated benefits and limitations are highlighted. PMID- 16372558 TI - Key issues in setting up and running a nurse-led cardiology clinic. AB - This article discusses the key issues involved in setting up and running an effective nurse-led cardiology clinic. These include: developing clear aims and objectives for the service, techniques to reduce resistance from colleagues who are affected by the change in service provision, professional development, audit and evaluation. PMID- 16372559 TI - Familial hypercholesterolaemia: clinical features and management. AB - Coronary heart disease is the leading cause of mortality and morbidity in the UK, for which hypercholesterolaemia is a risk factor. This article concentrates on the management of patients with familial hypercholesterolaemia (FH). It discusses FH, how it is diagnosed, and the treatments that are available. PMID- 16372560 TI - Neurological assessment. PMID- 16372562 TI - Rethink Christmas. PMID- 16372563 TI - Time is not on my side. PMID- 16372564 TI - Mentoring learners in practice: No. 16. Promoting health assessment learning. PMID- 16372565 TI - Water resources management in soft drink industry-water use and wastewater generation. AB - Water is used in most process industries for a wide range of applications. Processes and systems using water today are being subjected to increasingly stringent environmental regulations on effluents and there is growing demand for fresh water. These changes have increased the need for better water management and wastewater minimisation. In Morocco, water use in the food and drink industry is extensive at approximately 24 million m3 per year including 14% of drinking water in 1994. This study was conducted in a carbonate soft drink industry plant, during two years, 2001 and 2002. We have investigated the state of consumption and use of fresh water and the generation of the effluent in the factory. The aim of the study is to identify potential opportunities for reducing fresh water intake and minimising wastewater production by studying the posibility of reuse, recycling and treatment. PMID- 16372566 TI - Treatment of petrochemical wastewater by rotating biological contactor. AB - A laboratory scale study has been conducted to assess the efficiency of rotating biological contactor (RBC) to treat the synthetic wastewater from a petrochemical industry producing acrylonitrile. The attached biomass was acclimatised by gradually increasing cyanide concentration from 5 to 40 mg l(-1) with simultaneous increase in the concentration of chemical oxygen demand (COD) and ammonium nitrogen (NH4+-N). During acclimatization COD removal varied between 80 88 percent (%) while cyanide removal was more than 99%. The RBC was operated at varying hydraulic loading rates from 0.011 to 0.027 m3 m(-2) d(-1). The performance of the RBC was monitored for various parameters like COD, 5-day biochemical oxygen demand at 20-degree centigrade (degrees C) [BOD5], cyanide, ammonium nitrogen etc., for all hydraulic loadings. At all hydraulic loadings the cyanide removal remained more than 99%. The removals obtained in terms of percentage, for cyanide, COD, BOD5 and NH4+-N were greater than (>) 99, 95.2, 99.1 and 77, respectively at hydraulic loading of 0.011 m3 m(-2) d(-1). The effect of substrate/cyanide ratio on the performance of the process at five different ratios, 100/1, 80/1, 60/1, 40/1 and 20/1, showed more than 99% cyanide removal at ratio 20/1. The effect of COD/nitrogen (N) ratio was studied at four different ratios, 12/1, 10/1, 8/1 and 6/1, showed cyanide removal remained unaffected. The variation of biomass concentration within the system was also studied. PMID- 16372567 TI - Application of aerobic granular sludge in polishing the UASB effluent. AB - Stable aerobic granules were successfully developed in a sequencing batch reactor (SBR) fed with glucose and sodium acetate as the main carbon sources, and then they were acclimatized to polish the effluent from an up-flow anaerobic sludge blanket (UASB) reactor. It was found that more than 95% chemical oxygen demand (COD) and 85% suspended solids (SS) in the UASB effluent could be removed by an aerobic granular SBR as a post treatment unit. The performance of the SBR was stable over the whole experimental period. At an organic loading rate (OLR) of 4.5 to 5.4 g l(-1) d(-1), the acclimatized aerobic granules had an average diameter of 3.5 to 6.0 mm and a minimal settling velocity of 72 m h(-1). The biomass concentration in the SBR was as high as 8.4 g (dry weight) l(-1). This study showed for the first time that aerobic granular sludge in SBR would be an effective means to polish the effluent from UASB reactor. PMID- 16372568 TI - Simulation of pulp mill wastewater recycling after tertiary treatment. AB - The aim of this work is to study the possibilities of effluent recycling in a bleached Kraft pulp mill, for a better water management. To avoid problems associated with effluent recycling (corrosion, odors, loss in pulp and paper quality), wastewaters have to be treated before recycling. This study is particularly focused on organic matter removal. Several treatments are applied on a biological secondary effluent: adsorption on activated carbon, coagulation with ferric chloride or alum sulfate, precipitation with lime, ozonation and catalytic ozonation. These techniques are compared in terms of COD (Chemical Oxygen Demand) removal. Catalytic ozonation is finally chosen as the most effective solution to achieve 50% of COD removal in the effluent. The characteristics of the effluent treated according to this technique are then used to simulate the impact of its reuse in the process for pulp production. The study is focused on the changes in these parameters in the various stages of bleaching and final washing when water is replaced by the wastewater treated or directly issued from the wastewater treatment plant. The simulation demonstrates the need of a tertiary treatment to eliminate COD in order to avoid possible reactant overconsumption and decrease in pulp brightness. Chloride and sulfate ions which could cause corrosion should also be removed. PMID- 16372569 TI - Heavy metal removal from aqueous solution in sequential fluidized-bed reactors. AB - In this study, the sequential fluidized bed reactors (FBRs), were used to remove heavy metals including, Cu, Pb, and Ni, from synthetic wastewater. Heavy metals were removed through crystallization of metal carbonate and hydroxide precipitates on the surface of sand grains. The results showed that the influent metal concentration limits in the sequential FBRs were higher than those in the vertical FBRs. The removal efficiency for Cu, Pb, and Ni reached 96%, 93%, and 98% when the influent concentrations were 250 mg l(-1), 130 mg l(-1) and 130 mg l(-1), respectively. The pH value in the effluent of the FBR ranged from 8.7 to 9.1. The amount of metal coated onto the sand surface was determined and it was found that most of the metal ions were collected in the first reactor. The mechanism of heavy metal removal in the sequential FBRs concluded crystallization and filtration. PMID- 16372570 TI - Treatment of wastewater containing a large amount of suspended solids by a novel multi-staged UASB reactor. AB - Treatment of artificial wastewater containing a large amount of suspended solids comprised of soybean processing waste and pig fodder was studied using a novel multi-staged upflow anaerobic sludge blanket reactor. The reactor consisted of three compartments, each containing a gas solid separator. The wastewater had chemical oxygen demand of approximately 21600 mg l(-1), suspended solids of 12800 mg l(-1), and an ammonia concentration of 945 mg l(-1). A continuous experiment without effluent circulation showed that the multi-staged reactor was not that effective for the treatment of wastewater containing a large amount of suspended solids. However, operation of the reactor with circulation of effluent enabled the reactor to achieve organic removal of 85% and approximately 70% methane conversion at loading rates of between 4.0 to 5.4 kg-chemical oxygen demand per cubic meter per day, meaning that the reactor was more effective when effluent was circulated. Morphological investigation revealed that the crude fiber in the sludge was partially degraded and that it had many small depressions on its surface. Evolved biogas may have become caught in these depressions of the fibers and caused washout of the sludge. PMID- 16372571 TI - Startup of pilot-scale aerobic granular sludge reactor by stored granules. AB - The startup of a pilot-scale aerobic granular sludge reactor was investigated by seeding with 4-month stored aerobic granules. Two liters of granules were inoculated into the reactor (5.9% of reactor volume), which gave a biomass concentration of 1.03 g l(-1). Experimental results showed that seeding granules could be successfully maintained in the reactor. The microbial activity of seeding granules could be fully recovered to that of fresh granules after 2 days of operation, and new granules started to grow after day 5. Newly developed aerobic granules at stable period had similar size and morphology as seeding granules, and a biomass concentration of 6.0 g l(-1) was achieved in the reactor. The experiment demonstrated for the first time that stored aerobic granules could be used for easy and quick startup of aerobic granular sludge reactor. PMID- 16372572 TI - Competitive and hindering effects of natural organic matter on the adsorption of MTBE onto activated carbons and zeolites. AB - Equilibrium and kinetic adsorption of methyl tert-butyl ether (MTBE) onto three coal-based activated carbons, one coconut-based activated carbon, and two zeolites are elucidated in this study. Natural organic matter (NOM) and MTBE competed for the adsorption of activated carbons to different extents. The ideal adsorbed solution theory (IAST) combined with the equivalent background compound (EBC) model can adequately describe the NOM competition and predict the isotherms of MTBE onto the activated carbons. No competitive adsorption was observed for one of the zeolites, mordenite, due to the molecular effect. Besides, the aperture size, and the SiO2/Al2O3 ratio of the zeolite may also play an important role in the adsorption of MTBE from the aqueous phase. The surface diffusion model accurately simulated the transport of MTBE within the adsorbents employed in different water matrices. For all the activated carbons tested, the surface diffusivity of MTBE in natural water was nearly equal to that in deionized water, indicating that no apparently hindering effect occurs. A much slower adsorption kinetic of mordenite in natural water was observed since the opening apertures on mordenite may be appreciably hindered and blocked by NOM. PMID- 16372573 TI - Effect of sulfidogenic and methanogenic inhibitors on reductive dehalogenation of 2-chlorophenol. AB - The potential for reductive dehalogenation of 2-CP in anaerobic batch cultures of fresh-water digested sludge under sulfidogenic and methanogenic conditions was investigated in the presence or absence of respective inhibitors: molybdate and BESA at various concentrations (0 to 10 mM). Triplicate cultures (50% vol/vol) were set-up under an atmosphere of 20% CO2 and 80% N2 in 160 ml serum bottles using anaerobic digester sludge and a mineral medium containing 0.1% yeast extract. The dehalogenation of 2-CP, as well as methanogenesis, occurred at the same rate in the presence or absence of sulphate. Sulphate reduction did not inhibit 2-CP degrading populations. The presence of BESA--a known inhibitor of methane producers partially inhibited methanogenesis and slowed 2-CP dehalogenation at even 1 mM concentration with phenol and acetate accumulation in the cultures. The accumulation was proportional to the increase in concentration of BESA in the system. Molybdate on the other hand completely inhibited both sulphate reduction and 2-CP dehalogenation at a concentration of 10 mM. The dehalogenation of 2-CP continued in the presence of 1 mM molybdate even after the cessation of sulphate reduction indicating that sulphate-reducing bacteria were not directly involved in the dehalogenation of 2-CP in this study. Inhibition of 2-CP dehalogenation and sulphate reduction along with accumulation of propionate at 10 mM molybdate in the cultures strongly suggests that the dehalogenation of 2 CP was more directly linked to syntrophic activity of the mixed culture compared to sulphate reduction. PMID- 16372574 TI - Control strategies for a combined anaerobic (UASB)-aerobic (Activated Sludge) wastewater treatment system. AB - This paper presents a comparative analysis among four control strategies for the control of a combined anaerobic-aerobic wastewater treatment configuration. The anaerobic stage is an Upflow Anaerobic Sludge Blanket (UASB) reactor, whereas the aerobic stage consists of the Activated Sludge (AS) process. The control variables are solids concentration in the effluent and sludge wastage rate. The proposed control strategy is considered the most reliable among them. It is based on two cascaded PI-controllers for the solids concentration in the effluent and a look-up table for the sludge wastage rate control. Experimental and simulated results are presented. PMID- 16372575 TI - Equilibrium and kinetics studies for As(III) adsorption on activated alumina and activated carbon. AB - Equilibrium adsorption isotherms of As(III) were obtained from the experimentally generated data on activated alumina and activated carbon at temperature 303 K and pH 7.4. The isotherm for As(III) on activated alumina was typically of Brunauer Type-I but in the case of activated carbon equilibrium capacity seems to be increasing with increase in concentration. Langmuir isotherm model best represents the experimental data for As(III) in activated alumina and Freundlich model in activated carbon. Model parameters such as saturation capacity of adsorbent qs, Langmuir constant b, and Freundlich constants K and n were determined. Saturation adsorption capacity of As(III) in activated carbon was much higher than that in activated alumina. Overall effective mass transfer coefficient k was determined using uptake rate method. It is observed that k was independent of concentration of As(III). Values of k for both the adsorbents are of the order of 10(-2) min(-1). PMID- 16372576 TI - The use of pulsed ultrasound technology to improve environmental remediation: a comparative study. AB - Relatively little is known about the effects of pulsed ultrasound on the facilitation of chemical reactivity and remediation. Previous studies have indicated that sonochemistry using pulses may be either more or less effective than continuous wave (CW) ultrasonic irradiation. However, because of the time modulated nature of the pulses used in these studies, less acoustic energy in general was transmitted to the solutions compared to CW sonication. The effectiveness of ultrasound when the pulse is adjusted so that the same amount of acoustic energy is input compared to continuous irradiation over a given time has not been previously explored. In this study we have embarked on a comparison of the efficacy of power-modulated pulsed (PMP) sonochemistry with more traditional time-modulated pulsing. As a prototypical reaction, we have explored the effects of pulse type on the degradation of acid orange, a common industrial colorant. An increase in the degradation rate by a factor of three was observed using PMP ultrasound compared with continuous irradiation under conditions of equivalent acoustic input power, while the use of time-modulated pulsed ultrasound from a commercially available direct-immersion (DI) horn-type sonicator exhibited a rate decrease compared to CW sonication. Possible mechanisms for the enhancement are discussed. PMID- 16372577 TI - Closer to the gold standard: an appraisal of formulae available in Italy for use in formula-fed infants. AB - Infant formulae are the only alternatives to breast milk for infants who are unable to continue breastfeeding through the first year of life. They aim to provide formula-fed infants with the same structural and functional benefits observed in breastfed infants. To achieve this, bioactive nutrients have been added to infant formulae in recent years: long-chain polyunsaturated fatty acids for neurodevelopment; probiotics and prebiotics for local gastrointestinal defence; and nucleotides for promoting the immune response. Changes in protein quantity and quality allow infant formulae to achieve a balance between providing the correct plasma amino acid profile and reducing the protein intake, which could prevent obesity in later life. Hydrolysed proteins may help prevent atopic disorders. Many short-term trials have been published but long-term follow-up data are needed in infants who have been fed the newer infant formulae, to fully understand the role of bioactive nutrients. PMID- 16372578 TI - Effects of trimetazidine on lipid peroxidation, antioxidant enzyme activities and plasma brain natriuretic peptide levels in patients with chronic cor pulmonale. AB - An oxidant/antioxidant imbalance in favour of oxidants appears to occur in chronic cor pulmonale (CCP). Oxidative stress could also be a critical event in the pathogenesis of this condition. Trimetazidine (TMZ) has antioxidant properties and may affect the utilization of oxygen radicals. We investigated the effect of TMZ (20 mg three times daily, orally) on activities of erythrocyte malondialdehyde (MDA) and catalase (eCAT), erythrocyte and plasma glutathione peroxidase (GSH-Px) and plasma superoxide dismutase (pSOD) in CCP patients. We also assessed changes in plasma levels of brain natriuretic peptide (BNP) with TMZ therapy. Sixty CCP patients with significantly higher MDA and markedly lower pSOD, eCAT and GSH-Px (erythrocyte) activities than 24 healthy controls were randomly allocated to receive routine treatment or routine treatment plus TMZ. After 3 months' therapy, greater pSOD, eCAT and GSH-Px (erythrocyte and plasma) activities and lower MDA activity were found with TMZ treatment compared with routine treatment. Plasma BNP levels were significantly lower in TMZ-treated patients and higher in the routine treatment group than in the control group. TMZ improved antioxidant levels, decreased oxidative stress and decreased plasma BNP levels in CCP patients. PMID- 16372579 TI - Ambulatory blood pressure comparison of the anti-hypertensive efficacy of fixed combinations of irbesartan/hydrochlorothiazide and losartan/hydrochlorothiazide in patients with mild-to-moderate hypertension. AB - This study examined whether the greater anti-hypertensive efficacy of irbesartan monotherapy over losartan monotherapy extends to the respective fixed-dose combinations with hydrochlorothiazide (HCTZ) in patients with mild-to-moderate hypertension. Patients were treated with either irbesartan 150 mg/HCTZ 12.5 mg or losartan 50 mg/HCTZ 12.5 mg over a 4-week period. Twenty-four hour daytime and night-time mean blood pressure (BP), BP load and duration of action were assessed using ambulatory BP monitoring. Both treatment regimens significantly reduced BP from baseline for all efficacy variables assessed. A significant difference was noted in adjusted mean changes from baseline in 24-h ambulatory diastolic BP with irbesartan/HCTZ versus losartan/HCTZ. Reduction in diastolic load was significantly greater with irbesartan/HCTZ than with losartan/HCTZ as was mean ambulatory systolic BP during the last 4 h of the dosing interval. Both regimens were well tolerated, with no significant differences in terms of adverse event profile observed. Irbesartan 150 mg/HCTZ 12.5 mg resulted in greater reductions in ambulatory BP than losartan 50 mg/HCTZ 12.5 mg. PMID- 16372580 TI - Is the remarkable contradiction between histology and 14C urea breath test in the detection of Helicobacter pylori due to false-negative histology or false positive 14C urea breath test? AB - We assessed the diagnostic value of the 14C urea breath test (UBT) in the detection of Helicobacter pylori compared with histology and the rapid urease test (RUT). The study included 68 patients (22 men and 46 women) with dyspeptic symptoms. H. pylori status was evaluated by 14C UBT, RUT and histology. Sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy were determined for 14C UBT and for RUT. Histology revealed dense yeast like micro-organisms in the biopsy specimens in all patients with false-positive results by 14C UBT (n = 8), a significantly higher proportion than in patients with negative 14C UBT (five of 31). The low specificity of the H. pylori 14C UBT should not be neglected by accepting histology results as false-negative. Gastric mucosal colonization by yeast-like micro-organisms with urease activity can account for the high frequency of false-positive results for 14C UBT. PMID- 16372581 TI - Seroprevalence of hepatitis C virus infection and evaluation of serum aminotransferase levels among haemodialysis patients in Izmir, Turkey. AB - The seroprevalence of hepatitis C virus (HCV) infection was investigated among haemodialysis (HD) patients. Mean serum aminotransferase levels were also compared over 3 months in HCV-seropositive patients with and without viraemia, as well as in HCV-seronegative HD patients and HCV-seropositive, non-uraemic, viraemic patients. Seroprevalence of HCV infection was 19% among the 437 HD patients tested. Of the 61 HD HCV-seropositive, hepatotoxic medication- and alcohol-free patients, 38 (62%) were found to be viraemic, using quantitative HCV RNA, on at least one occasion. Mean serum aminotransferase levels were significantly higher in viraemic HD patients (compared with non-viraemic patients), suggesting that HCV-RNA positivity is an important predictor of increased enzyme activity in these patients. As expected, aminotransferase levels in HCV-seropositive HD patients tended to be lower than levels in HCV seropositive non-uraemic patients. PMID- 16372582 TI - DNA vaccine against hamster oral papillomavirus-associated oral cancer. AB - Previously we developed a carcinogenesis model involving the combination of 9,10 dimethyl-1,2-benzanthracene (DMBA) application with physical wounding of hamster lingual mucosa. The presence of a novel hamster oral papillomavirus (HOPV) was demonstrated and its genome sequenced. In the present study, this HOPV hamster model was used to test whether vaccination with the L1 gene could prevent the development of oral carcinoma. DNA plasmids encoding the L1 gene or the vector alone were injected intramuscularly into 20 vaccinated and 20 control hamsters, respectively. The lingual tips of the hamsters were painted with DMBA for 8 weeks. A portion of the lingual tips was excised, and the tips were then painted daily with DMBA until the animals were killed 13 days later. All control hamsters developed lingual carcinoma, whereas 12 of the L1-vaccinated hamsters showed no lesions. These results suggest that immunization with L1 DNA vaccines may prevent the development of papillomavirus-associated oral cancer. PMID- 16372583 TI - Amyloid beta protein deposition in osteopetrotic (op/op) mice is reduced by injections of macrophage colony stimulating factor. AB - The deposition of amyloid beta (Abeta) protein is a neuropathological change that characterizes Alzheimer's disease. Animals with the osteopetrosis (op/op) mutation suffer from a general skeletal sclerosis, a significantly reduced number of macrophages and osteoclasts in various tissues, and have no systemic macrophage colony stimulating factor (M-CSF). This study examined the effect that M-CSF injections had on Abeta deposition and microglial cell distribution in the brains of normal and op/op mice. Abeta-positive plaques were detected in the cerebral cortex of op/op mice, but not in normal mice. M-CSF reduced the numbers of Abeta-positive plaques in op/op mice. The microglial cell population was reduced in op/op mice compared with normal mice, and M-CSF increased the numbers to 65.8% of that observed in normal mice. Our results suggest that a clearer understanding of the role that microglial cells play in Abeta deposition may help determine the mechanisms involved in the pathogenesis of Alzheimer's disease. PMID- 16372584 TI - Enhanced expression of the local haematopoietic bone marrow renin-angiotensin system in polycythemia rubra vera. AB - Local bone marrow (BM) renin-angiotensin system (RAS) affects physiological and pathological haematopoiesis, including erythropoiesis. In this study, quantitative expression of the messenger RNAs of the major RAS components- angiotensin-converting enzyme (CD143), renin and angiotensinogen--were measured in BM samples by quantitative real-time polymerase chain reaction, to evaluate the activity of local BM RAS in polycythemia rubra vera (PV) in comparison with normal erythropoiesis. The presence of CD143 was also investigated in the same BM samples by flow cytometry. Increased local synthesis of the major RAS components has been identified by demonstrating corresponding mRNAs in the BM of the patients with PV. Our findings indicate up-regulation of local BM RAS, together with down-regulation of the cell surface angiotensin-converting enzyme receptors, in the autonomous neoplastic clonal erythropoiesis of PV. PMID- 16372585 TI - Usefulness of sparfloxacin against Chlamydia pneumoniae infection in patients with bronchial asthma. AB - The efficacy of sparfloxacin (SPFX) for the control of bronchial asthma was evaluated in 26 patients with suspected Chlamydia pneumoniae infection. Patients were randomly allocated to receive SPFX 200 mg/day (n = 14) or control treatment (n = 12) for 21 days. Significant improvements in serum C-reactive protein levels, and significant decreases in peripheral eosinophil counts, serum eosinophil cationic protein (ECP) and sputum ECP were observed in the SPFX treated group at day 21. SPFX-treated patients also had a significantly reduced frequency of asthma symptoms, reduced inhalant beta2-stimulant use, and significant increases in morning peak expiratory flow. At the end of the study, C. pneumoniae was undetectable in two SPFX-treated patients who underwent polymerase chain reaction testing, but one control patient who was tested still had detectable levels of C. pneumoniae. These results suggest that SPFX could be used to control bronchial asthma in patients with suspected persistent C. pneumoniae infection. PMID- 16372586 TI - The telmisartan renoprotective study from incipient nephropathy to overt nephropathy--rationale, study design, treatment plan and baseline characteristics of the incipient to overt: angiotensin II receptor blocker, telmisartan, Investigation on Type 2 Diabetic Nephropathy (INNOVATION) Study. AB - We planned the INNOVATION study to determine whether telmisartan, an angiotensin 2-receptor blocker, delays the progression of renal disease from incipient nephropathy to overt nephropathy in hypertensive or normotensive Japanese patients with type 2 diabetes mellitus. The INNOVATION study is a randomized, double-blind, placebo-controlled trial. Eligible patients must have incipient nephropathy (defined as a urinary albumin to creatinine ratio of 100-300 mg/g creatinine) and a serum creatinine concentration of < 1.5 mg/dl for men and < 1.3 mg/dl for women. Patients who need treatment with angiotensin II receptor blockers or angiotensin-converting enzyme inhibitors are excluded. Eligible patients are randomly assigned to three groups: telmisartan titrated to 40 mg; telmisartan titrated to 80 mg; or placebo. The primary endpoint is the time from baseline visit to first detection of overt nephropathy (defined by a urinary albumin to creatinine ratio that is > 300 mg/g creatinine and 30% higher than the baseline on at least two consecutive visits). A total of 1855 patients have been enrolled from 160 study centres. In 527 randomized patients (28.4% of the enrolled patients), mean (SD) urinary albumin to creatinine ratio and serum creatinine concentration at baseline were 173.3 (47.2) mg/g creatinine and 0.78 (0.19) mg/dl. Sixty-eight per cent of the patients had hypertension at baseline. Mean (SD) systolic and diastolic blood pressures at baseline were 137.1 (14.6) and 77.5 (10.3) mmHg. The INNOVATION study will determine whether telmisartan, an angiotensin II receptor blocker, provides clinical benefits in hypertensive or normotensive patients with diabetes mellitus and diabetic nephropathy. PMID- 16372587 TI - Oxidant/antioxidant parameters and their relationship with chemotherapy in Hodgkin's lymphoma. AB - This study investigated changing levels of serum oxidant/antioxidant with chemotherapy and their relation to treatment in 34 Hodgkin's lymphoma patients. The patient population consisted of 19 males and 15 females. Mean age was 30.41 +/- 12.08 years. All patients received the adriamycin, bleomycin, vincristine and dexamethasone (ABVD) treatment protocol. Blood samples were taken before treatment, and on days 1 and 7 during treatment for measurement of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), malondialdehyde (MDA), nitric oxide (NO) and enzyme activities. After ABVD treatment, mean free radical levels were increased and antioxidant levels were significantly decreased in the serum. ABVD treatment results in an increase of free radical levels and a decrease of antioxidant levels in the serum of patients with Hodgkin's lymphoma. PMID- 16372588 TI - Effect of pre-operative angiotensin-converting enzyme inhibitors on haemodynamic parameters and vasoconstrictor requirements in patients undergoing off-pump coronary artery bypass surgery. AB - The effects of pre-operative angiotensin-converting enzyme inhibitor (ACEI) treatment on haemodynamic status and vasoconstrictor requirements during off-pump coronary artery bypass surgery (OPCAB) were studied. Eighty patients selected for OPCAB were divided into those who had been treated with ACEIs for 4 weeks or more pre-operatively (ACEI group) (n = 43) and those who had not been treated with ACEIs (control group) (n = 37). Noradrenaline was infused during the operation when the mean systemic arterial pressure (SAP) fell below 60 mmHg. No significant differences in the haemodynamic parameters measured were detected between the two groups, except for cardiac output, which was found to be significantly greater in the control group. During anastomosis of the obtuse marginal branch of the left circumflex artery (OM), a significantly larger amount of noradrenaline was required by the ACEI group compared with the control group. In conclusion, pre operative treatment with ACEIs significantly increased the amount of vasoconstrictor necessary to maintain the target SAP during OM anastomosis during OPCAB. PMID- 16372589 TI - Primary hydatid disease in femoral muscles. AB - Usually, intramuscular hydatid cysts are secondary, resulting from the spread cysts from other areas either spontaneously or after operations for hydatidosis in other regions. We present an unusual case of a primary hydatid cyst found in the left thigh of a 35-year-old woman, presenting as an enlarging soft-tissue tumour. Ultrasound, computed tomography and magnetic resonance imaging examinations revealed a multilocular intramuscular cyst in the anterior aspect of her left thigh, and no disease at any other location. We removed the entire cyst surgically, and macroscopic and microscopic histopathological examinations confirmed the diagnosis of muscular hydatidosis. Three years after the operation there had been no recurrence. In regions where hydatidosis is endemic, a tumour in any part of the body should be considered a hydatid cyst until proven otherwise. PMID- 16372590 TI - Retrograde hydrostatic irrigation enema-induced perforation of the sigmoid colon in a chronic renal failure patient before colonoscopy. AB - We present a rare case of colon perforation caused by hydrostatic irrigation enema in a patient with chronic renal failure. A 76-year-old woman was admitted to our hospital because of an exacerbation of lumbar pain and increased difficulty in walking. She had a medical history of traumatic neck pain and chronic lower back pain, which had been treated with non-steroidal anti inflammatory drugs (NSAIDs) for 8 years. On admission, the C-reactive protein level was 6.8 mg/dl, so we planned to do a colonoscopy to determine the cause of inflammation. The patient developed abdominal pain approximately 3.5 h after a pre-procedural enema was administered. An emergency operation was performed and a small perforation was found in the sigmoid colon. We conclude that the cause of the colon perforation was a combination of the use of a hydrostatic retrograde irrigation enema in a patient with chronic renal failure who had been treated with long-term NSAIDs. PMID- 16372591 TI - Cerebral venous thrombosis initially considered as a complication of spinal epidural anaesthesia. AB - Cerebral venous thrombosis is an unusual condition characterized by headache, nausea, vomiting, focal deficits and epileptic seizures. In this case report we describe a patient who presented with headache and focal motor deficits after an uneventful Caesarean section, performed using combined spinal-epidural anaesthesia. Magnetic resonance imaging was performed because of the pronounced neurological symptoms, and a diagnosis of cerebral venous thrombosis was made. The patient was treated with anticoagulant agents and made a complete recovery. This case emphasizes the importance of considering cerebral venous thrombosis in the differential diagnosis of headache in the post-partum period prior to instituting conventional therapy. PMID- 16372592 TI - Concepts of renormalization in physics. AB - A non technical introduction to the concept of renormalization is given, with an emphasis on the energy scale dependence in the description of a physical system. We first describe the idea of scale dependence in the study of a ferromagnetic phase transition, and then show how similar ideas appear in particle physics. This short review is written for non-particle physicists and/or students aiming at studying particle physics. PMID- 16372593 TI - Molecular engineering of surfaces using self-assembled monolayers. AB - The self-assembly of molecules into structurally organized monolayers (SAMs) uses the flexibility of organic chemistry and coordination chemistry to generate well defined, synthetic surfaces with known molecular and macroscopic properties. The process of designing monolayers with a specified structure gives a high level of control over the molecular-level composition in the direction perpendicular to a surface; soft lithographic technique gives useful (if lower) resolution in the plane of the surface. Alkanethiolates adsorbed on gold, silver, mercury, palladium and platinum are currently the best-defined systems of SAMs. They provide substrates for a number of applications-from studies of wetting and electron transport to patterns for growing mammalian cells. SAMs have made organic surfaces a central part of surface science. Understanding the principles by which they form, and connecting molecular-level structure with macroscopic properties, opens a wide range of areas to study and exploitation. PMID- 16372594 TI - An introduction to RNA-mediated gene silencing. AB - Careful analysis of cases where introduction of additional copies of endogenous genes caused coordinate silencing of both the transgene and the endogenous gene laid the ground work for the discovery of RNA-mediated silencing. Silencing begins with the expression and recognition of double-stranded RNA, which is cleaved into short RNAs that recognize, by complementarity, sequences that are targets for down regulation. An RNA target can be regarded (post-transcriptional gene silencing), but the small RNAs can also direct the sequence-specific modification of DNA and chromatin. RNA-mediated gene silencing in eukaryotes may have originated as surveillance mechanism to protect the organism from transposable elements and viruses and then evolved to specify chromosomal modifications and to regulate expression of a significant fraction of endogenous genes by microRNAs. This review seeks to furnish the student and non-expert with some idea of how RNA-mediated silencing was discovered and a broad overview of the present state of knowledge. PMID- 16372595 TI - Diabetes: lessons for midwives. AB - This study is larger than any other in describing pregnancy outcomes for women with pre-gestational diabetes. From the report, the three main messages for midwives are: Women with type 2 diabetes have a risk of poor outcome similar to those with type 1 diabetes. The same support and management of women with type 1 diabetes should be applied to women with type 2 diabetes pre-conceptionally and throughout the pregnancy and neonatal period. Pre-pregnancy care and advice should be made more flexible and advertised well in a variety of settings where diabetic women of childbearing age are most likely to see it. Women with diabetes should be encouraged and supported to breastfeed their babies from birth by giving them an understanding of the general and specific benefits this will provide. PMID- 16372596 TI - Don't play down role of emergency obstetric care. PMID- 16372597 TI - Pregnant adolescents: who cares? PMID- 16372598 TI - Delivering care over a cuppa. PMID- 16372599 TI - Domestic violence: learning to ask the question. PMID- 16372600 TI - Giving birth in the dust: the Palestinian experience. PMID- 16372601 TI - Midwifery basics: complications in pregnancy (3). Neonatal resuscitation. PMID- 16372602 TI - Maternal breastfeeding positions: have we got it right? (2). PMID- 16372603 TI - Dealing with antenatal domestic abuse in Wales. PMID- 16372604 TI - The birth of water embolism. PMID- 16372605 TI - The Christmas crib. PMID- 16372606 TI - T4 rectal cancer: analysis of patient outcome after surgical excision. AB - Locally advanced rectal cancer dictates a major surgical undertaking, which includes en bloc resection of the rectum and all involved organs. The aim of this study was to evaluate patient outcome and compare multimodality treatment options after various surgical approaches from one institution for T4 rectal cancer. A retrospective chart review identified 24 patients who were operated on for advanced primary rectal cancer invading adjacent structures (T4) over a 5(1/2) year period. The types of treatment and outcome were analyzed. From these 24 patients, the most frequently involved organ was the bladder (33%). A total of 16 patients underwent chemoradiotherapy. There were 12 complications (50%), the most common being wound infection (33% of complications, 17% overall). Nine patients had nodal disease. Disease-free survival was 54 per cent, and overall survival was 75 per cent. However, disease-free survival in node-negative patients was 67 per cent versus 33 per cent in node-positive individuals. Out of the six patients who died in this review, five (83%) received chemoradiotherapy. Operations for advanced primary rectal cancer with involvement of adjacent organs are major procedures associated with high morbidity. Patients without nodal disease may have long-term survival despite the locally advanced tumor. Interestingly, neoadjuvant therapy, adjuvant, or both, did not increase survival. PMID- 16372607 TI - The results of superficial femoral, popliteal, and tibial artery stenting for peripheral vascular occlusive disease. AB - The purpose of this study was to evaluate the patentcy rates and physiological effectiveness of angioplasty and stenting as a primary therapy for superficial femoral (SFA), popliteal (POP), and tibial (TIB) arterial occlusive disease. Seventy-eight patients had stents placed in the infra-inguinal vessels between January 1, 2001, and July 31, 2004. We collected data on patient demographics, symptoms, as well as pre- and postprocedure ankle-brachial index (ABI) and angiographic findings. Patency rates at 6 months, 12 months, and 24 months were analyzed by life table methods. Thirty-one men and 47 women had a mean age of 68 years (range 36-94 years). Risk factors included diabetes in 50 per cent, hypertension in 79 per cent, smoking in 41 per cent, and end-stage renal disease in 10 per cent. The indications for intervention were claudication in 52 per cent and limb salvage in 48 per cent of patients. Stents were placed in the SFA in 54 patients (69%), in the POP in 15 patients (18%), and in the TIB artery in 6 patients (8%). Average follow-up was 11.2 months. The mean postprocedural increase in ABI was 0.29. The 6-month, 1-year, and 2-year primary patency rates were 83 per cent, 58 per cent, and 47 per cent, respectively. Limb salvage was achieved in 66 per cent of patients treated for limb-threatening ischemia. There was one major and three minor complications. Stenting of the infra-inguinal vessels has a low morbidity, high success rate, and acceptable patency and limb salvage rates. PMID- 16372608 TI - Unexpected findings during thyroid surgery in a regional community hospital: a 5 year experience of 738 consecutive cases. AB - Unexpected findings during thyroid surgery in a nonuniversity setting have rarely been reported in large series. Our goal was to describe the unexpected findings during thyroid surgery in a busy regional community hospital. All thyroid operations conducted by the teaching staff at Greenville Memorial Hospital, a 735 bed nonuniversity regional hospital, from December 1998 through December 2003 were reviewed. Pre- and post-operative diagnoses, surgical procedure, and specimen histopathology were examined. Unexpected findings were defined as either thyroid pathology not anticipated based on preoperative diagnosis or as unsuspected nonthyroidal disease found during cervicotomy. During the 5-year study period, 738 patients presented with thyroid disease requiring surgery. Incidental thyroid cancer was discovered in 28 cases (3.8%), the predominance being papillary microcarcinoma. Synchronous benign thyroid disease, separate from the indication from surgery, was observed in 56 patients (7.6%). Forty patients had unexpected nodular goiter and 16 had lymphocytic thyroiditis. Primary hyperparathyroidism was observed in 33 patients (4.5%). Both solitary adenomas (22 cases) and multigland parathyroid disease (11 cases) were seen. Unexpected nonendocrine findings were less common, including solitary cases of large cell carcinoma, metastatic endometrial carcinoma, and tracheal duplication (bronchogenic cyst). In conclusion, unexpected findings during thyroid surgery at a busy community hospital are fairly common. Indeed, an unanticipated finding is encountered in one out of seven operations on the thyroid gland. Although most are of unclear clinical significance, there is a surprisingly high incidence of hyperparathyroidism. This underscores the need for preoperative screening, as the "thyroid patient" may also be the "parathyroid patient." PMID- 16372609 TI - Water-soluble upper GI based on clinical findings is reliable to detect anastomotic leaks after laparoscopic gastric bypass. AB - Anastomotic leak after laparoscopic Roux-en-Y gastric bypass (LGB) is a major complication that must be recognized and treated early for best results. There is controversy in the literature regarding the reliability of upper GI series (UGI) in diagnosing leaks. LGB was performed in patients meeting NIH criteria for the surgical treatment of morbid obesity. All leaks identified at the time of surgery were repaired with suture and retested. Drains were placed at the surgeon's discretion. Postoperatively, UGI was performed by an experienced radiologist if there was a clinical suspicion of leak. From September 2001 until October 2004, a total of 553 patients (age 40.4 +/- 9.2 years, BMI 48.6 +/- 7.2) underwent LGB at UAB. Seventy-eight per cent (431 of 553) of patients had no clinical evidence suggesting anastomotic leak and were managed expectantly. Twenty-two per cent (122 of 553) of patients met at least one inclusion criteria for leak and underwent UGI. Four of 122 patients (3.2%) had a leak, two from anastomosis and two from the perforation of the stapled end of the Roux limb. No patient returned to the operating room without a positive UGI. High clinical suspicion and selectively performed UGI based on clinical evidence is reliable in detecting leaks. PMID- 16372610 TI - Impact of length of stay on functional outcomes of TBI patients. AB - The purpose of this study was to compare the functional outcomes of two groups of patients with traumatic brain injury (TBI) with attention to the impact of reduced length of stay (LOS) in the trauma center (TC) and rehabilitation hospital (RH). From 1991 to 1994, 55 patients, Group 1, with serious TBI (Abbreviated Injury Scale score > or = 3) were admitted to a level 1 TC and subsequently transferred to a comprehensive inpatient RH. These results have been previously published. From 1996 to 2002, 64 similarly injured patients, Group 2, received inpatient care at the same TC and RH. These patients had a marked decrease in length of stay. Functional Independence Measures (FIM) were obtained at admission (Adm), discharge (D/C), and at 1 year follow-up for both groups. The average length of stay at the TC dropped from 36 days in Group 1 to 26 days in Group 2. In addition, the average length of stay at the RH dropped from 46 days (Group 1) to 25 days (Group 2); overall, an average reduction of 31 days of inpatient care. Group 2 had significantly lower FIM scores at the time of RH discharge for self-care, locomotion, and mobility compared to Group 1. At the 1 year follow-up, however, there were no significant differences between Groups 1 and 2 in these FIM scores. FIM scores at 1 year were higher in Group 2 for communication (90% vs 71%) and social cognition (77% vs 49%) compared to Group 1. Over one-fourth of each group returned to work by the 1 year follow-up. Socially disruptive behavior occurred at least weekly in 28 per cent (Group 1) and 23 per cent (Group 2) of patients. The outcome for serious TBI is better than generally perceived. Reduction of inpatient LOS did not adversely affect the ultimate functional outcome. The decreased LOS placed a greater demand on outpatient rehabilitative services as well as a greater burden on the family of the brain injured patient. PMID- 16372611 TI - Laparoscopic ventral hernia repair: postoperative antibiotics decrease incidence of seroma-related cellulitis. AB - Seroma formation has been documented as a common complication in laparoscopic ventral herniorraphy. However, there are no recent studies documenting the incidence of or protective strategies against seroma-related cellulitis. The purpose of this study was to evaluate 65 laparoscopic ventral herniorraphies and to determine if seroma-related cellulitis can be prevented by the routine use of postoperative prophylactic antibiotics. A retrospective case review of 65 laparoscopic ventral herniorraphies was done at our institution from February 2002 to January 2004. All were performed using either Gore-Tex DualMesh or Bard Composix mesh and performed under the direct supervision of a single surgeon. Twenty patients received only preoperative third-generation cephalosporins or fluoroquinolones. All other patients received either 7 days of postoperative oral cephalosporins or fluoroquinolones in addition to preoperative antibiotics. Sixty five patients underwent laparoscopic ventral hernia repair. There were 45 patients in the postoperative antibiotic group and 20 patients in the preoperative-only antibiotic group. Twenty-one patients developed seromas. Twelve of these developed cellulitis. The rates of seroma formation were similar in the two groups with 30 per cent in the preoperative only group and 33 per cent in the postoperative antibiotic group. However, 100 per cent of the seromas in the preoperative antibiotic group developed seroma-related cellulitis. Only 40 per cent of seromas in the postoperative antibiotic group developed cellulitis. In addition, two seromas in the preoperative antibiotics-only group progressed to frank mesh infection necessitating operative removal. There were no complications related to antibiotic administration. Laparoscopic ventral hernia repair is a safe and effective procedure. Our seroma rate is 30 per cent and compares equally with prior reported studies. Seroma-related cellulitis is a common problem that can lead to mesh infection, postoperative morbidity, and further need for operative care. The administration of 7 days of postoperative prophylactic antibiotics appears to be a safe and effective means to limit seroma-related cellulitis. PMID- 16372612 TI - All-terrain vehicle injuries: are they dangerous? A 6-year experience at a level I trauma center after legislative regulations expired. AB - All-terrain vehicles (ATVs) have increased in popularity and sales since 1971. This rise in popularity led to an increase in injuries resulting in voluntary industry rider safety regulations in 1988, which expired without renewal in 1998. Our purpose was twofold, to determine the incidence and severity of ATV injuries in our patient population and what, if any impact the safety regulations had. To further characterize the risk of ATV use, we compared them to a vehicle generally recognized as dangerous, the motorcycle (MC). Our trauma registry was reviewed from January 1998 through August 2004 for ATV or MC injured. Data collected included age, gender, mortality, Injury Severity Score (ISS), helmet use, and injury distribution. These were compared to our data from the decade of regulation. There were 352 MC and 221 ATV patients. ATV injured demonstrated a higher proportion of pediatric and female patients (P < 0.001 and P < 0.01, respectively), a decrease in helmet use (8.6% vs 64.7%, P < 0.001), and increased closed head injuries (CHI) (54.2% vs 44.9%, P < 0.05) compared with MC injured. ISS and mortality were similar. The average number of patients from 1988 to 1998 was 6.9/yr compared to 31.6/yr (P < 0.001) during 1998-2004 with equal ISS. Our data show that there has been a dramatic and progressive increase in the number of ATV crashes since expiration of industry regulations. ATVs are as dangerous as MCs based on patient ISS and mortality. There are significantly more children and women injured on ATVs. The lower rate of helmet use in ATVs may account for the significantly greater incidence of CHI. These data mandate the need for injury prevention efforts for ATV riders, in particular children, through increased public awareness and new legislation. PMID- 16372613 TI - Is outcome after blunt splenic injury in adults better in high-volume trauma centers? AB - An association between outcome and case volume has been demonstrated for selected complex operations. The relationship between trauma center volume and patient outcome has also been examined, but no clear consensus has been established. The American College of Surgeons (ACS) has published recommendations on optimal trauma center volume for level 1 designation. We examined whether this volume criteria was associated with outcome differences for the treatment of adult blunt splenic injuries. Using a state trauma database, ACS criteria were used to stratify trauma centers into high-volume centers (>240 patients with Injury Severity Score >15 per year) or low-volume centers, and outcome was evaluated. There were 1,829 patients treated at high-volume centers and 1,040 patients treated at low-volume centers. There was no difference in age, gender, emergency department pulse, emergency department systolic blood pressure, or overall mortality between high- and low-volume centers. Patients at low-volume centers were more likely to be treated operatively, but the overall success rate of nonoperative management between high- and low-volume centers was similar. These data suggest that ACS criteria for trauma centers level designation are not associated with differences in outcome in the treatment of adult blunt splenic injuries in this regional trauma system. PMID- 16372614 TI - Amelioration of gastroesophageal reflux symptoms following Roux-en-Y gastric bypass for clinically significant obesity. AB - Symptoms of gastroesophageal reflux disease (GERD) are frequent in patients with clinically significant obesity and are reported to improve after Roux-en-Y gastric bypass (RYGB). The purpose of this study is to determine timing and duration of improvement of GERD symptoms in patients undergoing RYGB. Prospectively collected data from patients who underwent RYGB from January 1998 to August 2004 were analyzed. Patients answered a standardized questionnaire pre- and postoperatively inquiring about frequency of GERD symptoms (none, one episode/ week, one episode/day, more than one episode/day) and medication use. Of 606 patients undergoing RYGB, 239 patients (39%) reported GERD symptoms preoperatively (mean age 43 +/- 1 years; body mass index 51 +/- 1 kg/m2). Of these, 89 per cent of patients reported improved at 3 months post-op and 94 per cent of patients 9 months post-op (P < 0.001). Medication usage decreased from 30 per cent to 3 per cent by 3 months and 5 per cent beyond 9 months (P < 0.001). Percentage of excess weight loss was 18 +/- 1 per cent and 75 +/- 2 per cent at 3 and 9 months, respectively. Symptoms of GERD significantly improve and use of antireflux medications is reduced after RYGB independent of weight loss. RYGB may be the treatment of choice for GERD in obese patients. PMID- 16372615 TI - Selective use of intraoperative touch prep analysis of sentinel nodes in breast cancer. AB - Imprint cytology (touch prep) is often used for intraoperative examination of sentinel nodes in breast cancer. This allows axillary lymph node dissection (ALND) to be performed immediately for tumor-positive nodes. We evaluated the accuracy of touch prep examination of sentinel nodes and its role in the surgical treatment of breast cancer. We analyzed 169 breast cancer patients who underwent 170 lymphatic mapping procedures with intraoperative touch prep examination. Results from the touch prep were correlated with histopathology and clinical variables. There were 115 true-negative, 35 true-positive, 15 false-negative, and 5 false-positive results. Touch prep had a sensitivity of 70 per cent and specificity of 96 per cent. Positive predictive value, negative predictive value, and diagnostic accuracy were all 88 per cent. The false-negative rate was 30 per cent and correlated with the size of the nodal metastasis and number of involved nodes, but not other patient factors. Touch prep is useful for the evaluation of sentinel nodes in breast cancer, but it has a lower sensitivity than initially reported, particularly in patients with micrometastases. False positive results occur, although they may be reduced after experience with the technique. We recommend that suspicious findings on touch prep should be confirmed by frozen section and that ALND only be performed for histologically documented metastases. We currently perform touch prep only in patients who are at high risk of nodal metastasis or will undergo mastectomy. This improves operative efficiency and limits the impact of false positive and negative results without dramatically increasing the number of patients who require a second surgical procedure. PMID- 16372616 TI - Obesity status and use of general surgery resources. AB - Obesity is a rapidly growing epidemic. This study assesses the impact of obesity on surgeon workload for general surgical services. A retrospective study of patients undergoing cholecystectomy, unilateral mastectomy, and colectomy between January 2000 and December 2003 was undertaken. Obesity was defined as body mass index > or = 30. The proportion of obese patients was compared to the 2002 BRFSS obesity prevalence data for Alabama. Data were adjusted to control for potential confounders. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. A total of 1,385 patients were included in analysis. The prevalence of obesity in the study population was 35.5 per cent compared to the statewide prevalence of 25.2 per cent (OR = 1.73, 95% CI = 1.51, 1.98). These data were stratified by procedure, age, and gender. The cholecystectomy group had a significantly higher proportion of obese for all age groups and female gender. The mastectomy group had a higher proportion of obese in the 45-64 age group. The stratified colectomy group did not reach statistical significance. There was no evidence of referral bias to explain these findings. This study demonstrates there is a greater use of general surgery services, particularly cholecystectomy and mastectomy, in obese patients than predicted by the prevalence of obesity in the population. PMID- 16372617 TI - Personal satisfaction and mentorship are critical factors for today's resident surgeons to seek surgical training. AB - The specific aim of this study was to summarize the viewpoints of the Resident and Associate Society of the American College of Surgeons (RAS-ACS) membership regarding current training and quality of life-related issues prior to implementation of the new duty-hour guidelines. The goal was to gain insight of the members that may be useful to recruit and guide the future training of surgical residents. An Internet-based survey was developed to evaluate the viewpoints of RAS-ACS. The survey was administered by Esurveymaker.com via the ACS Web page from 2000 to 2003. RAS-ACS member participation was voluntary and anonymous. Analyses were performed to determine the frequency of response for each survey item. Two hundred thirty-five members completed the survey representing 5 per cent of RAS-ACS. Eighty-four per cent were general surgery residents. Personal satisfaction (64%) and mentorship (49%) were top factors for respondents to pursue surgical training; discussion with colleagues and future income was less important. Forty-five per cent reported that job performance was their most important concern during residency. A rewarding surgical career and family life were ranked as the most important expectations. Eighty-six per cent reported that they were satisfied with their residency, and 66 per cent reported that work hours should be limited. Personal satisfaction and mentorship were critical factors for members of the RAS-ACS to seek surgical training. Although most of the members report that work hours should be limited, an overwhelming majority reports satisfaction with surgical training prior to institution of the new duty-hour guidelines. Further emphasis on mentorship and work-hour reform may be beneficial in recruiting medical students into surgical residencies. PMID- 16372618 TI - Hyperacute abdominal compartment syndrome: an unrecognized complication of massive intraoperative resuscitation for extra-abdominal injuries. AB - Primary and secondary abdominal compartment syndrome (ACS) are well-recognized entities after trauma. The current study describes a "hyperacute" form of secondary ACS (HACS) that develops intraoperatively while repair of extra abdominal injuries is being carried out simultaneous with massive resuscitation for shock caused by those injuries. The charts of patients requiring abdominal decompression (AD) for HACS at time of extra-abdominal surgery at our level I trauma center were reviewed. The following data was gathered: age, Injury Severity Score (ISS), mechanism, resuscitation details, time to AD, time to abdominal closure, and outcome. All continuous data are presented as mean +/- standard error of mean. Hemodynamic and ventilatory data pre- and post-AD was compared using paired t test with significance set at P < 0.05. Five (0.13%) of 3,750 trauma admissions developed HACS during the 15-month study period ending February 2004. Mean age was 32 +/- 7 years, and mean ISS was 19 +/- 2. Four of five patients arrived in hemorrhagic shock (blunt subclavian artery injury, 1; chest gunshot, 1; gunshot to brachial artery, 1; stab transection of femoral vessels, 1) and were immediately operated upon. One of five patients (70% burn) developed HACS during burn wound excision on day 2. HACS developed after massive crystalloid (15 +/- 1.7 L) and blood (11 +/- 0.4 units) resuscitation during prolonged surgery (4.8 +/- 0.8 hours). Pre- versus post-AD comparisons revealed significant (P < 0.05) improvements in mean arterial pressure (55 +/- 6 vs 88 +/- 3 mm Hg), peak airway pressure (44 +/- 5 vs 31 +/- 2 mm Hg), tidal volume (432 +/ 96 vs 758 +/- 93 mL), arterial pH (7.16 +/- 0.0 vs 7.26 +/- 0.04), and PaCO2 (52 +/- 6 vs 45 +/- 6 mm Hg). There was no mortality among the group, and all patients underwent abdominal closure by fascial reapproximation in 2-5 days. Two (40%) of the five patients required extremity fasciotomy for compartment syndrome. HACS is a rare complication of massive resuscitation for extra abdominal injuries. It should be considered in such patients in the face of unexplained hemodynamic and/or ventilatory decompensation. Prompt AD is life saving. Early abdominal closure is usually possible. Vigilance for compartment syndromes elsewhere in the body is warranted in any patient with HACS.) PMID- 16372619 TI - Secondary abdominal compartment syndrome: risk factors and outcomes. AB - Secondary abdominal compartment syndrome (ACS), defined as intra-abdominal hypertension with associated pulmonary, renal, or hemodynamic compromise in the absence of preceding abdominal operation or injury, can markedly increase surgical morbidity and mortality. We performed a retrospective chart review of the physiologic parameters and outcomes of 10 patients with secondary ACS. Ten patients developed secondary ACS after aggressive resuscitation, at an average of 20.2 hours. Four of the patients sustained burns greater than 40 per cent, three of the patients had penetrating extremity trauma, one patient had blunt abdominal trauma, one patient was struck by lightning, and one patient developed a retroperitoneal bleed while on heparin. The average bladder pressure was 40.6. The average volume given in the first 24 hours was 33,001 cc (range, 12,400 to 69,000). The average base deficit at admission was -12 (range, +1 to -25). Seven of the 10 patients had decreased urine output. Nine of the 10 patients had decreased tidal volumes on pressure control ventilation. All 10 patients were hypotensive, with 7 of the 10 requiring vasopressors. Overall mortality was 60 per cent, with 43 per cent mortality for those decompressed. Prompt recognition and treatment are mandatory for survival of ACS. We recommend routine bladder pressure monitoring for patients with ongoing resuscitation greater than 500 cc/hr. PMID- 16372620 TI - Protocol for bedside laparotomy in trauma and emergency general surgery: a low return to the operating room. AB - Bedside laparotomy (BSL) was introduced as a heroic procedure in trauma patients too unstable for safe transport to the operating room (OR). We hypothesize a BSL protocol would maintain patient safety while reducing OR use. Patients were prospectively entered into a BSL protocol from July 2002 to June 2003 and retrospectively reviewed. Protocol indications for BSL were abdominal compartment syndrome, decompensation due to hemorrhage, washout/closure, and sepsis in a patient too unstable for safe transport to the OR. Primary outcomes were mortality, emergent return to OR, and primary fascial closure (PFC). Trauma operating room charges and OR time were analyzed. One hundred thirty-three BSL were performed on 60 patients with an overall mortality of 23.3 per cent (14/60). There was an average of 2.2 BSL per patient (range 1-8). Indications for BSL were 1) explore/washout (n = 100, 75.2%), 2) decompression (n = 14, 10.5%), 3) infection/abscess (n = 12, 9.0%), 4) hemorrhage (n = 7, 5.3%). Five of 133 BSL (5.8%) were emergently returned to the OR because of perforation or compromised bowel. Trauma OR charges were dollar 5,300 per cases with 2.12 hours per cases. The protocol standardized the conduct of BSL procedure to allow for a low return to OR rate of 5.8 per cent and had an overall in-hospital mortality rate of 23.3 per cent. Primary fascial closure of the abdomen had a significantly reduced hospital stay. BSL allowed trauma OR charges of dollar 5,300 per cases with 2.12 hours per cases savings. PMID- 16372621 TI - BSN by 2010: a California initiative. PMID- 16372623 TI - Fundamentals of grant writing: lessons learned from the process. PMID- 16372622 TI - Review of leadership and management literature. PMID- 16372624 TI - Latex allergy: implications for nurse educators. PMID- 16372625 TI - Peer learning: a clinical teaching strategy to promote active learning. PMID- 16372626 TI - The seven habits for highly effective curriculum revision. PMID- 16372627 TI - Teaching student parents. PMID- 16372628 TI - Transforming pathophysiology instruction through narrative pedagogy and Socratic questioning. AB - Pathophysiology, heavily content driven, has typically been taught through the use of traditional behavioral pedagogy and a reliance on the formal lecture. The author describes the limitations of this approach to teaching pathophysiology and describes the use of narrative pedagogy and Socratic questioning as alternative methods of instruction to augment lecture methods. Specific strategies for transforming traditional classroom teaching by using Socratic questions in a pathophysiology course for nurse practitioners are described. Student and faculty reactions to the initial efforts to transform pathophysiology instruction are also described. PMID- 16372629 TI - Developing a community nursing center. AB - With the move to community-based nursing education, schools of nursing are seeking ways to implement high-quality clinical experiences for undergraduate and graduate students in partnership with the community. The author presents ten tips for the development of a community nursing center that accomplishes this goal. PMID- 16372630 TI - Who wants to be a nurse? PMID- 16372631 TI - Mind-mapped care plans: a remarkable alternative to traditional nursing care plans. AB - Nursing care plans have helped students learn problem solving for nursing practice, but creativity and the interrelationship of patient problems are not stimulated by their linear nature. Joining mind mapping with care planning forces connections, engages whole brain thinking, and stimulates creativity. The authors describe mind mapping, infusion of mind-mapped care plans into the curriculum, the teaching/learning process of mapped care plans, and the positive outcomes of mind mapping nursing care plans. PMID- 16372632 TI - Promoting clinical scholarship through scholarly writing. AB - To promote clinical scholarship, critical thinking, problem-solving ability, and effective writing and research skills in their students, faculty replaced their major care plan with a capstone scholarly paper. The authors discuss how faculty, who serve as mentors, guide their students through the development of the scholarly paper. PMID- 16372633 TI - Faculty practice in a small liberal arts college. AB - Small liberal arts colleges are challenged to find ways to participate in faculty practice. With limited access to large healthcare centers, small liberal arts colleges must create innovative faculty practice models. The author shares one college's experience in faculty practice that can be used by others, detailing practice development, funding, developmental challenges, and future goals. PMID- 16372634 TI - Fostering interaction through multimedia. AB - Multimedia is a vehicle for fostering interaction and reinforcing the connection between curriculum and learning. The author discusses ways to energize, reshape, and enrich educational experiences by integrating multimedia into instruction. Examples of teaching-learning strategies and processes that promote learner content, learner-learner, and learner-instructor interactions are offered to demonstrate how multimedia use can enhance student self-reflection, problem solving, critical analysis, cultural sensitivity, and acquisition and development of new knowledge. PMID- 16372635 TI - It takes two to deliver. PMID- 16372636 TI - A pediatric learning experience: respite care for families with children with special needs. AB - Respite care provides temporary relief from the demands of constant caregiving for primary care givers. While better known in geriatrics, respite care can also promote family well-being and enhance the coping abilities of parents with children with special needs. The authors describe how a respite care service and a department of nursing collaborated to provide respite care to parents of children with special needs, thus providing a much needed service in the community as well as a unique clinical experience for nursing students. PMID- 16372637 TI - Enhancing nursing students' experiences with health promotion and preventative care. PMID- 16372638 TI - Journal of Undergraduate Nursing Scholarship. PMID- 16372639 TI - Use of primary trait analysis to develop: research prospectus guidelines. PMID- 16372640 TI - Polanyi's theory of knowledge: the mandate for hands-on clinical learning. PMID- 16372641 TI - Faculty Development: an approach to scholarship. AB - This consultation to promote scholarship resulted in the development of two projects, a study of the relationship between power and teaching-learning and the integration of holistic concepts into the curriculum. These projects were developed because the consultant was able to promote faculty discussion of organizational issues that had been felt but not verbalized. The discussion of these concerns permitted insight into group dynamics and clarified the concerns that had hampered effective group functioning and the pursuit of scholarship. The faculty became empowered to more effectively communicate with the administration faculty values and the importance of creating an environment to enhance scholarly activities. The consultant on scholarship was the catalyst who promoted the change in attitudes and activities. PMID- 16372642 TI - Helping expand nurse practitioner students' clinical skills repertoire: learning minor procedures. AB - This course is immensely popular with students. Many express a sense of accomplishment in knowing that they have performed a specific procedure at least once before facing the requirement in the clinical setting. Students who complete the course have a firm foundation in minor procedures and are knowledgeable about indications, contraindications, and methods to perform the procedure before facing that first patient in the clinical setting. They have also accumulated a set of procedures, patient discharge instructions, and a bibliography that can be reviewed before performing a procedure for the first time in the clinical setting. Combining clinical hands-on skills with the experience of writing a step by-step procedure reinforces learning and is a valuable skill that can be used in clinical practice after graduation. Although many of the students will only apply a portion of the skills they learn in this course, they express a significant boost in self-confidence, a decrease in anxiety level, a sense of accomplishment in their skills, and a definite edge in securing a nurse practitioner position in a competitive healthcare marketplace. PMID- 16372643 TI - Using Web CT to determine competency in medication dosage calculation for nursing students. PMID- 16372644 TI - Mistakes, I've made a few: student medication errors. PMID- 16372645 TI - Simulated clinical scenarios: faculty-student collaboration. PMID- 16372646 TI - Decreasing anxiety for the nursing student entering the acute-care clinical setting. PMID- 16372647 TI - Predicting NCLEX-PN performance for practical nursing students. AB - Success on the licensure examination, NCLEX-PN, is a fundamental outcome criterion for practical nursing programs as well as a state requirement for professional practice. Nurse educators and administrators have an interest in identifying factors that predict success on the licensure examination. This study examined demographic, pre-admission, and programmatic variables to determine their predictive value for NCLEX-PN success in a statewide 2-year college system. PMID- 16372648 TI - Involving nursing students in continuous improvement projects. AB - Reducing medication errors is a topic of national concern and action. Nursing students participated in a healthcare organization's continuous quality improvement project targeting patient safety. Students were actively involved in chart review and became acutely aware of safety issues related to medication administration, order transcription and implementation, and documentation. Both the students and the hospital realized expected and unexpected benefits. PMID- 16372649 TI - Providing leadership opportunities for associate degree nursing students. PMID- 16372650 TI - Building a case for promotion of clinical faculty. AB - A newly created and approved clinical faculty track was without a well-defined path to promotion. The authors describe how the school of nursing clinical track faculty integrated the 4-part model of scholarship defined by Boyer and Rice into the traditional promotion triad of research, teaching, and service. The authors' challenge was defining promotion guidelines to achieve respect, reward, and promotion. PMID- 16372651 TI - Weaving undergraduate research into practice-based experiences. AB - Most nursing education programs fail to link research to practice-based courses, thus contributing to the research-to-practice-gap. To better prepare graduates for evidence-based practice, a project that linked research to practice-based learning was developed. The author discusses how the Collaborative Learning Project (CLP) was designed and implemented, the learning activities that linked research to practice-based experiences in an adult health course, and the outcomes of the strategy. PMID- 16372652 TI - Implementing a nursing program via live interactive video: lessons learned. AB - Flexibility in scheduling and sensitivity to meeting the educational needs of today's diverse student population is a must in nursing education. The authors present an overview of how a college of nursing implemented an entire generic BSN program via live interactive television for nursing students 45 miles from the main campus. The authors describe the experience of initiating the program, facilitating faculty and student adaptation to the challenges of live interactive video education, and a summary of the first five years. PMID- 16372653 TI - Online critical thinking: a case study analysis. AB - Educating nursing students using Internet technology, in conjunction with case studies, is rewarding for both faculty and the learner The authors describe an Internet journal discussion format used as an adjunct assignment in a senior clinical nursing course. Adapting a traditional teaching strategy such as case studies proved successful online. Results showed students were able to think critically about a clinical situation and recognize the importance of peer communication. PMID- 16372654 TI - Leadership and empowerment: the value of the National Student Nurses Association for beginning students. AB - Leadership development has long been emphasized in nursing curricula, but how can it be effectively taught and learned? The authors discuss how beginning student experiences with the National Student Nurses Association (NSNA) promotes colleagueship, research utilization, policy development, political activity, and advocacy. Student reflections on their experience illuminate how they were empowered to effect change and understand their social responsibility. PMID- 16372655 TI - Physician advisors can help case managers do their job better. PMID- 16372656 TI - Physician-case manager team cuts LOS, denials. PMID- 16372658 TI - Get involved in your insurance contracts. PMID- 16372657 TI - Don't rely on Condition Code 44 to fix mistakes. PMID- 16372659 TI - Hospital's DM program cuts admissions, ED visits. PMID- 16372660 TI - Lean production efforts help save dollar 7.5M in 1 year. PMID- 16372661 TI - JCAHO to look closely at patient handoffs. PMID- 16372662 TI - System achieves 'right patient, right level of care'. PMID- 16372664 TI - Progress towards wild poliovirus containment in Russian Federation. PMID- 16372663 TI - CHF project aims to bridge gap between providers. PMID- 16372665 TI - Ten things you need to know about pandemic influenza (update of 14 October 2005). PMID- 16372666 TI - Health conditions for travellers to Saudi Arabia pilgrimage to Mecca (Hajj). PMID- 16372667 TI - [Iacob Iacobovici]. PMID- 16372668 TI - [Central pancreatectomy--indications, technique, outcomes]. AB - Central pancreatectomy is a conservative resectional procedure indicated for benign and low malignant tumors located in the neck and/or body of the pancreas. We report our experience on 5 patients and some considerations about this surgical technique from medical literature. PMID- 16372669 TI - [Conversion in laparoscopic cholecystectomy]. AB - Laparoscopic cholecystectomy (LC) has become the gold standard in the treatment of symptomatic cholelithiasis. Some patients require conversion to open surgery and several preoperative variables have been identified as risk factors that are helpful in predicting the probability of conversion. The aim of this study was to analyze the factors that make LC difficult and determine conversion to open approach: Our study includes: 6985 cases which underwent LC and 1430 cases with open cholecystectomy, between March 1993 and April 2005 in our clinic of general surgery. The overall conversion rate was 5.1% (deliberate conversion--299 cases, conversion of necessity--62 cases). The conversion rate has decreased from 17.5% in 1993 to 3.2% in recent years. The most conversion happen after a simple inspection or a minimal dissection caused by the existence of perforation (105 cases), the discovery of a difficult anatomic situation (63 cases) or of another pathology (14 cases); more rarely, the conversion was necessary in the principal time, doing to hemorrhage (26 cases), impossible dissection (41 cases), visceral injury (1 case) or even at the end of the operation, doing to hemorrhage, loss piece or stone (10 cases), and other situations (101 cases). Significant predictors of conversion were acute cholecystitis , choledocholithiasis, past history of acute cholecystitis, male gender, gall bladder wall thickness exceeding 6 mm. In conclusion, based on our experience, we suggest limiting OC to patients with proven contraindications to LC (i.e., Mirizzi syndrome or systemic illness incompatible with general anesthesia or pneumoperitoneum), attempting LC in all other cases. Decision to convert to open approach is a proven of surgical maturity. Conversion must be decided from the beginning, in the moment of the recognition of a difficult situation and not after the occurrence of a complication. PMID- 16372670 TI - [Surgical treatment in severe acute pancreatitis. Last 15 years of experience in Emergency County Hospital of Baia Mare]. AB - The aim of this study is to investigate the particular course of the patients operated for severe acute pancreatitis in a period of 15 years in surgical department of Emergency County Hospital of Baia Mare. Medical records of 202 patients admitted and operated for severe acute pancreatitis, were studied. Follow-up parameters were: age, gender, etiology, moment of operation, the type of operations and postoperative evolution of this patients. In the group of deceased patients alcoholic etiology of pancreatitis was prevailing. Almost a half of patients were operated in the first day of admission. A high number of patients were operated for diagnosis of acute abdomen with intention of exploratory laparotomy. In the last years, besides the usual closed drainage, open drainage and planning drainage were performed. Postoperative mortality is still high. The diagnose of severe acute pancreatitis is difficult in emergency. Global mortality in pancreatitis remains high, especially in the period of enzymatic shock, and is correlated with masculine gender, alcoholic etiology and somewhat with precocity of operation. PMID- 16372671 TI - Expression of HLA-DR antigen and characterization of the lymphocytic infiltrate in normal mucosa, tubulo-villous adenoma and invasive carcinoma of the colon. AB - Prognosis of colonic carcinoma is poor. The two most important factors having the greatest effect on survival are pathologic stage of disease and histologic grade of the tumor. Our study points towards the value of HLA-DR antigen in the prognosis of colonic carcinoma. We studied 31 cases of normal colonic mucosa, 12 cases of tubulo-villous adenoma, and 39 cases of invasive carcinoma for the detection of HLA-DR monoclonal antigen. Yet, we investigated the association of HLA-DR and DQ genes and adenoma and carcinoma by PCR. We also studied the T helper (TH) marker (CD4) in the lamina propria of the relevant cases, given that the dependence of immune responsiveness on the class II antigens reflects the central role of these molecules in presenting antigen to TH cells. HLA-DR was expressed in 20 of 31 normal colonic mucosa (64.5%), 4 of 12 adenomas (33.3%), and in 10 of 39 invasive carcinomas (25.6%). No significant correlation between HLA-DR and DQ genes and adenoma or cancer of the colon was found. CD4 was expressed in 9 of 31 normal colonic mucosa (29%), 5 of 12 adenomas (42%), and in 26 of 39 invasive carcinomas (67%). The results showed decreased expression of HLA-DR and increased expression of CD4 as the lesion progressed to malignancy. HLA-DR and DQ genes do not contribute to a susceptibility to adenoma or carcinoma. The immune attract mechanism by low HLA-DR signaling seems to be of minor importance in the malignant and metastatic potential of the colonic carcinoma. PMID- 16372672 TI - [The main utility of trans-anastomosis external drainage in the hydatid liver cysts treatment by peri-cystic-jejunostomy]. AB - We purpose to make a metanalysis of a external drainage and his role in the treatment of the hydatid liver cysts when it was made a anastomosis between peri cystic cavity and a isolated loop of jejunum. The paper watch 74 cases which were operated in "I. Juvara" surgical clinic and in which it was used the external drainage of peri-cystic cavity. The drainage roles consist in follow up of peri cystic cavity evolution, but, more than that, in early diagnosis and treatment of secondary infection of cavity, the main postoperative complication. The drainage excludes the possibility of unfavourable evolution of postoperative infection, in which case the result of operation may be compromised. PMID- 16372673 TI - [Liver trauma. Experience of Surgical Department, City Hospital Timisoara]. AB - The present paper analyzes the liver trauma, underlying efficient diagnostic methods and treatment approaches. Liver trauma is frequent in Surgical Emergency Units and it is a challenge for the surgeon who has to deal with diagnosis and treatment problems and also to treat the associated lesions. The study analyzes 49 cases of liver trauma admitted in Emergency Surgery Clinic from Timisoara City Hospital between January 1995 and April 2005. A very good trained team specialized in liver surgery but also a multidisciplinary team is very important in liver trauma. PMID- 16372674 TI - [Surgical methods of covering head soft tissues defects]. AB - Posttraumatic and post-excisional head soft tissue defects represent a challenge for the plastic surgeon due to the functional and esthetic impact of this body area. Choosing the simplest way of covering a defect is many times the most efficient way, but it must deliver a good tissue cover and their aspect and function must simulate the characteristics of that area. This article is proposing to be a brief description of all these procedures, informing and being necessary for every general surgeon. The methods of covering head soft tissues defects specific to plastic surgery will be presented in a ladder way from simple to complex, that means from the simple skin graft to island flaps and the tissues transfers. Choosing a right procedure refers to the etiopathogeny, and the length and width of the defect. We shall present advantages and disadvantages of each procedure, indications and their limits, giving specific clinical cases. We shall present the results for each type of procedure also, giving specific arguments for our option. We'll insist on the functional reconstruction of the lids, nose and cheek, as single defects and on the surgical limits which come upon very large soft tissue defects. Due to the complexity of the head structures, either single or complex, and to the functional and esthetic impact of these soft tissue defects, we consider this paper to be a real benefit. It is very important to choose the right procedure, considering all the factors who are implicated (sex, age, general condition, etiopathogeny, the length and width of the defect, previous treatments, patient wish, etc). PMID- 16372675 TI - [Small intestinal hemorrhage due to rare etiology. Diagnostic difficulties and therapeutic approaches]. AB - Understanding of the pathogenesis, diagnosis, and treatment of lower GI bleeding has drastically changed during the last 50 years, but it continues to be a frequent cause of hospital admission and also a factor in hospital morbidity and mortality. Acute lower gastrointestinal hemorrhage (ALGIH) represents only 20% of the GI bleeding and the small intestine is the site of hemorrhage in about 1% of cases. Although in the last four decades, diagnostic methods for locating the precise bleeding point improved greatly, still the adequate localization of the lesion is very difficult to achieve through algorithmic approaches. We performed a clinical study and we retrospectively analyzed 5 patients (mean age = 59,8 years, 1 female) who had a surgical intervention for acute lower GI hemorrhage in the last decade, in order to emphasize diagnosis difficulties when the bleeding arises from small lesions in the small bowel that is not easily accessible for direct visualisation. Surgery was the treatment of choice in every case consisting in segmental resections of the involved small bowel, along with viscerolysis and exploratory gastro-jejunostoma. The mortality rate was 20% (1 postoperative death of cardiac etiology). There have been no specific postoperative complications in the other four patients and a good outcome was reported. The diagnosis is particularly difficult and when colonic and upper gastrointestinal evaluations fail to identify a source of bleeding, a small intestinal source should be considered. We can conclude that the most important factor in the management of ALGIH is determination of specific localization of the lesion. PMID- 16372676 TI - [Subtotal abdomino-cervical esophagectomy (transhiatal, without thoracotomy, or Orringer's technique)]. PMID- 16372677 TI - [Double localization of pancreatic insulinoma. Diagnostic and therapeutic difficulties]. AB - Insulinomas are the most common cause of hypoglycemia resulting from endogenous hyperinsulinism. Because most of insulinomas are less than 2 cm in size and rarely they not may be visible by CT scan or transabdominal ultrasonography. Intraoperative ultrasonography may be a solution. Although as surgical method is preferred enucleation because operative time is shorter and easier and the low frequency postoperative complications, pancreaticoduodenectomy Whipple is indicated in selected cases. We report a case of double insulinoma located in the head of the pancreas in which the diagnosis and surgical treatment presented difficulties which determined a particular clinical evolution. PMID- 16372678 TI - [Crohn's disease. Clinical and therapeutical considerations]. AB - Crohn's disease is a chronic granulomatous inflammatory condition of the intestinal tract of unknown etiology. Most commonly the disease affects the small bowel, the colon and the rectum. The acute and aggressive forms can evolve fast, mimicking an acute surgical illness, requiring surgical intervention in emergency. Surgical therapeutical option, in this condition, must be determined strictly by establishing a correct intraoperative diagnosis, through macroscopic features and histologic evidence. Because it is an incurable disease with variable evolution, marked by recurrence, that involves repeated surgical intervention, the surgical treatment (often resection), must be most conservative from the small bowel. We present 3 cases of surgical interventions with emergency characteristics (bowel obstruction through fitobezoar, colonic tumors obstruction of colon splenic angle, urachal infected tumors). In these cases the diagnosis was established intraoperatively and the surgical intervention was adapted to the particular cases. PMID- 16372679 TI - [Aneurysm of the splenic artery]. AB - The work is presenting the case of a young patient with splenic aneurysm, a more and more frequent diagnosis in these last years. The peculiarity consist in the biphase and bi-directional evolution, first toward the gastric lumen, with hematemesis and consequently, after about a month, in the peritoneal cavity, with hemoperitoneum. The positive diagnosis and the final surgical solution was obtained after successive procedures, due to the lack of an appropriate technical equipment (angiographic exploration) due to the lack of the medical information at one time (inefficient anamnesis and the lack of the documentation) and finally, due to some elements that disturbed the surgical reasoning. PMID- 16372680 TI - [Abdominal compartment syndrome in multiple trauma patients with concomitant abdominal and head lesions --mechanisms and therapeutical aspects]. AB - The acute and persistent increase of the intra-abdominal pressure (IAP) above 25 cm H2O represents a pathologic entity called the abdominal compartment syndrome (ACS). This intra-abdominal high pressure is responsible of dysfunctions in cardiovascular, respiratory and renal systems and also dysfunctions in brain and abdomen. In polytrauma patients with combined head and abdominal lesions, increases of IAP are transmitted via thorax and superior venous caval system to brain generating intracranial hypertension, reducing cerebral blood flow (CBF) and cerebral perfusion pressure (CPP) thus worsening neurological status and increasing mortality. A better prognosis for this polytrauma patients is possible only by elucidating the mechanisms which produces the dysfunctions in ACS and by promoting an aggressive therapeutical surgical approach. PMID- 16372682 TI - How much should we inform our patients? PMID- 16372681 TI - [The 9th National Conference of the Bulgarian Society of Surgery ( October 6-8, 2005, Varna)]. PMID- 16372683 TI - Screen-film versus full-field digital mammography: comparison in breast cancer screening. PMID- 16372684 TI - A retrospective evaluation of patient acceptance of computed tomography colonography ("virtual colonoscopy") in comparison with conventional colonoscopy in an average risk screening population. AB - PURPOSE: To evaluate patient acceptance of computed tomography (CT) colonography compared with conventional colonoscopy by means of a self-assessed questionnaire. MATERIAL AND METHODS: Four-hundred-and-one patients with valid address information from our patient collective were preselected. Patient acceptance was evaluated retrospectively using a self-assessed questionnaire. The patients underwent CT colonography in our institution using 4x or 16x multi-detector row (MDR) CT. Two-hundred-and-forty-six patients returned a completed questionnaire, 157 of these indicating that they had undergone both virtual and conventional colonoscopy. RESULTS: One-hundred-and-twenty (76.4%) of the 157 patients would undergo another CT colonography if necessary, while only 14 patients would not. One-hundred-and-sixteen (73.9%) patients favored the actual examination procedure of CT colonography (P<0.0001), while only 6.4% preferred the conventional method. Preparation prior to CT colonography was experienced as more convenient than preparation prior to conventional colonoscopy (52.2% versus 14%). CONCLUSION: CT colonography was preferred in terms of both preparation and the actual procedure itself. PMID- 16372685 TI - Computed tomographic colonography: comparison of two workstations. AB - PURPOSE: To compare two commercially available computed tomography (CT) colonography systems with respect to interobserver variability, the influence of level of expertise, and the gradual reduction of reviewing time for each system. MATERIAL AND METHODS: Two residents and two radiologists using Siemens CTAPP Colography software and Viatronix V3D-Colon software reviewed supine and prone CT acquisitions from 24 patients in a primary 3D endoluminal view. The observers graded each case with respect to technical quality and diagnostic value, assessed the presence of pathology, and indicated the time spent on the viewing. RESULTS: Significant differences were found in technical quality (P < 0.001) and diagnostic value (P<0.001) depending on which system was used, with higher scores for the Viatronix software. The agreement between specialists tended to be higher than that between residents (kappa=0.63 (0.30-0.95) vs. kappa=0.51 (0.21-0.81)), and the residents gave significantly (P < 0.001) higher scores of technical quality. However, the level of expertise had no significant impact on the assessments. We noted extensive variability in pathological lesions found by the different observers. The number of findings did not differ between workstations, but the viewers tended to report larger polyp sizes with the Viatronix software. The time needed for viewing decreased significantly from the first to the last examination viewed by each observer. CONCLUSION: Both the evaluated systems present trustworthy images of the human colon, but in a primary 3D setting the Viatronix software is favored owing to the user-friendly interface, higher experienced technical quality, and better diagnostic value. PMID- 16372686 TI - Follow-up and final results of the Oslo I Study comparing screen-film mammography and full-field digital mammography with soft-copy reading. AB - PURPOSE: To compare cancer detection rates of screen-film (SFM) and full-field digital mammography (FFDM) with soft-copy reading in a screening program including the initial positive scores for interval cancers and cancers in the subsequent screening round, and to analyze the false-negative FFDM interpretations. MATERIAL AND METHODS: Using a paired study design, 3683 women underwent SFM and FFDM in a population-based screening program. Two standard views of each breast were acquired. The images were interpreted without previous films for comparison. Independent double reading using a 5-point rating scale for probability of cancer was used for each modality. An examination was defined as positive if at least one of the two independent readers scored 2 or higher on the 5-point rating scale. SFM-positive cases were discussed in a SFM consensus meeting and FFDM-positive cases in a separate FFDM consensus meeting before recall. The study population was followed for more than 2 years so that interval cancers and screen-detected cancers in the subsequent screening round could be included. Cancer detection rates were compared using the McNemar test for paired proportions. The kappa statistic and Wilcoxon signed-rank test for matched pairs were used for comparing rating scores. The reading time was recorded for all FFDM interpretations. RESULTS: A total of 31 cancers (detection rate 0.84%) were diagnosed initially, of which SFM detected 28 and FFDM 23 (McNemar test P=0.23, discordant pair 8 and 3). Two cancers with a positive score at initial SFM reading and three with a positive score at initial FFDM reading were dismissed at SFM and FFDM consensus meetings, respectively. The difference in cancer detection after recall (discordant pair 11 and 5) was not significant (McNemar test, P=0.21). Of the 10 interval cancers and 16 screen-detected cancers in the subsequent round, 3 had true-positive SFM scores while 4 had true-positive FFDM scores in the initial reading session. A total of 38 cancers therefore had a positive result at double reading at one or both modalities, 31 at SFM and 27 at FFDM (McNemar test, P=0.48). Comparison of SFM and FFDM interpretations using the mean score for each case revealed no statistically significant difference between the two modalities (Wilcoxon signed-rank test for matched pairs; P-value=0.228). Two initial round cancers (one tumor found incidentally at work-up for a mass proved to be a simple cyst with a positive score at FFDM but a negative score at SFM, and one tumor with positive score at SFM but negative score at FFDM due to positioning failure) were excluded from the further analysis. Excluding these two cancers from comparison, there were 31% (22 of 72) false-negative SFM and 47% (34 of 72) false-negative FFDM individual interpretations. The overall mean interpretation time for normal FFDM examinations was 45 s. For most false negative FFDM results, the reading time was shorter or longer than for normal examinations. The recorded FFDM interpretation time was noticeably short for several overlooked cancers manifesting as microcalcifications (ductal carcinoma in situ). CONCLUSION: There is no statistically significant difference in cancer detection rate between SFM and FFDM with soft-copy reading in a mammography screening program. Analysis of cancers missed at FFDM with soft-copy reading indicates that close attention has to be paid to systematic use of image display protocols. PMID- 16372687 TI - Ultrasound-guided core needle biopsy of non-palpable breast lesions: a prospective analysis in 204 cases. AB - PURPOSE: To assess the diagnostic value of ultrasound (US)-guided 14 G core needle breast biopsy in non-palpable suspicious breast lesions. MATERIAL AND METHODS: From August 1997 to April 2001, 198 patients with 204 suspicious non palpable breast lesions underwent US-guided large core needle biopsy. Biopsies were performed with a free-hand technique using US equipment with a 7.5 MHz linear-array transducer; a minimum of three cores were obtained from each lesion. Pathological findings in US-guided core biopsy were correlated to findings in subsequent surgery or long-term (more than 2 years) imaging follow-up. RESULTS: Among the 204 non-palpable breast lesions for which histopathological findings were obtained by US-guided core biopsy, 118 were malignant (114 carcinoma, 2 metastasis, 1 lymphoma, and 1 malignant phyllodes tumor) and 86 were benign (4 carcinoma and 82 benign lesions confirmed at surgery or after at least 2 years of follow-up). Sensitivity, specificity, positive predictive value, and negative predictive value for diagnosis of malignancy in our series were 97%, 100%, 100%, and 95%, respectively. Diagnostic yield with 1, 2, 3, and 4 specimens per lesion was 73.5%, 88%, 94%, and 97.5%, respectively. CONCLUSION: US-guided core needle biopsy is a sensitive percutaneous biopsy method for diagnosing non-palpable breast lesions. To achieve a high diagnostic yield, a minimum number of three cores per lesion is advisable. PMID- 16372688 TI - In situ central pulmonary artery thrombosis in primary pulmonary hypertension. AB - A rare case of extensive in situ central pulmonary artery thrombosis in primary pulmonary hypertension (PPH) is presented. The differentiation from chronic thromboembolic pulmonary arterial hypertension (CTEPH) is of paramount importance because of different therapeutic strategies. In this case, the presence of mural thrombus in the central pulmonary arteries on computed tomography made the distinction difficult. However, the possibility of in situ thrombosis was suggested on the basis of absence of other findings of CTEPH (abrupt narrowing/truncation of segmental arteries, variation in size of segmental vessels, arterial webs, mosaic attenuation, pulmonary infarcts, and dilated bronchial arteries), and this was confirmed on final pathology. PMID- 16372689 TI - Effect of informed consent for intravascular contrast material on the level of anxiety: how much information should be given? AB - PURPOSE: To compare the effect of two different informed consent forms on patients' anxiety level prior to intravenous contrast material (IVCM) injection. MATERIAL AND METHODS: Two different informed consent forms were randomly given to 265 consecutive patients referred for either intravenous pyelography or computed tomography requiring IVCM injections. Form 1 had brief information, including only the common risks and risk factors associated with IVCM, while Form 2 had more comprehensive information. Before being presented with the two different informed consent forms, 191 of the 265 patients filled out two other forms evaluating: 1: How they felt in that particular situation/moment (STAI-T); and 2: How they felt independently of the situation or condition at that moment (STAI S). After the patients were informed, the STAI-T (measuring how they felt in that particular situation) was filled out once more to see if the anxiety level had changed. RESULTS: The anxiety level for 88 patients receiving Form 1 decreased after they were given the informed consent (P=0.033). However, among the 103 patients receiving Form 2, the anxiety level showed a significant increase (P=0.001) compared to the values obtained before Form 2 was given. CONCLUSION: These results indicate that informed consent, including brief information about the risk factors and potential adverse reactions of IVCM, reduces anxiety level, while detailed information before the procedure increases the anxiety level. PMID- 16372690 TI - Reactive sclerosis: hyperintense appearance on T2-weighted magnetic resonance imaging. AB - Osteosclerosis is defined as increased density of bone on X-ray imaging studies. It is known that osteosclerosis appears hypointense on both T1- and T2-weighted magnetic resonance imaging sequences. In this review, we present our experience in various sclerotic skeletal pathologies that appear hyperintense on T2-weighted magnetic resonance imaging. We emphasize the possible pathophysiological mechanisms that may cause this appearance, such as bone marrow edema and/or composition of newly formed bone. PMID- 16372691 TI - Imaging techniques used in the diagnosis, staging, and follow-up of patients with myeloma. AB - Radiologists play a central role in the diagnosis, initial staging, follow-up, and restaging of patients with myeloma. This review article attempts to familiarize the reader with all the various types of myeloma, their imaging appearances and useful imaging strategies. The staging system for myeloma patients has been updated and now includes findings from advanced imaging modalities. Radiologists have a vast array of imaging modalities at their disposal to aid them in diagnosis, staging, and follow-up. Currently, conventional radiographic skeletal surveys, magnetic resonance imaging, and F-18 FDG PET/CT examinations are the most useful instruments. PMID- 16372692 TI - Computed tomography for evaluation of rotation dislocation of supracondylar elbow fractures in children. AB - We describe a simple method for evaluation rotation dislocation of supracondylar elbow fractures. PMID- 16372693 TI - Magnetic resonance imaging of anterior cruciate ligament tears: evaluation of standard orthogonal and tailored paracoronal images. AB - PURPOSE: To evaluate the three standard orthogonal imaging planes and a paracoronal imaging plane for anterior cruciate ligament (ACL) tears. MATERIAL AND METHODS: Ninety patients (91 knees; 29 F and 61 M) aged between 15 and 84 years (mean 36.9 +/- 16.4 years) underwent magnetic resonance imaging (MRI) of the knee prior to arthroscopy. At surgery, 32 knees had an intact ACL, 4 a partial tear, and 55 a complete ACL tear. In all patients, axial, sagittal, coronal, and paracoronal T2-weighted turbo-SE images were acquired. The ACL was classified as intact, partially, or completely torn. Partial and complete tears were combined for statistical evaluation. RESULTS: Partial ACL tears (four cases) were not correctly diagnosed at MRI except in one knee by one observer on coronal images. Sensitivity in detecting ACL tears was 95%/63% (reader1/reader2) in the axial, 93%/95% in the sagittal, 93%/86% in the coronal, and 100%/93% in the paracoronal plane. Specificity was 75%/81% in the axial, 72%/81% in the sagittal, 78%/94% in the coronal, and 78%/88% in the paracoronal plane. CONCLUSION: ACL tears can be diagnosed accurately with each of the standard orthogonal planes. Based on reader confidence and interobserver agreement paracoronal images may be useful in equivocal cases. PMID- 16372694 TI - Transient splenial lesion of corpus callosum associated with antiepileptic drug: conventional and diffusion-weighted magnetic resonance images. AB - Transient focal lesions of splenium of corpus callosum can be seen as a component of many central nervous system diseases, including antiepileptic drug toxicity. The conventional magnetic resonance (MR) findings of the disease are characteristic and include ovoid lesions with high signal intensity at T2 weighted MRI. Limited information exists about the diffusion-weighted MRI characteristics of these lesions vanishing completely after a period of time. We examined the conventional, FLAIR, and diffusion-weighted MR images of a patient complaining of depressive mood and anxiety disorder after 1 year receiving antiepileptic medication. PMID- 16372695 TI - Posterior cranial fossa crowdedness is related to age and sex: an magnetic resonance volumetric study. AB - PURPOSE: To measure 3-dimensional (3D) posterior cranial fossa (PCF) crowdedness and to evaluate the effect of age, sex, and body height on PCF. MATERIAL AND METHODS: Fifty-two healthy volunteers (24 M and 28 F; mean age 55.4 +/- 17.2 years; range 24-82 years) were recruited. Using a semi-automated magnetic resonance technique, we calculated a PCF crowdedness index (CI) as the ratio of hindbrain (HB) volume to PCF volume x 100% and correlated this index with age, sex, body height, and other crowdedness parameters. RESULTS: The mean PCF CI was 93.7 +/- 2.7%. Women had a more crowded PCF than men (95.0+1.7% versus 92.1 +/- 2.7%; P<0.001). PCF CI declined with age for both men (r = -0.61; P=0.002) and women (r = -0.68; P<0.001). The association with age--but not HB volume was maintained after we controlled for sex and body height. On multiple regression, both age and sex accounted for 57.5% of the PCF CI variance. CONCLUSION: Our study shows that PCF CI is associated with age and sex, and can therefore be used as a surrogate to assess hindbrain atrophy in a cross-sectional sample. Moreover, sex- and age-specific normal ranges may be needed to evaluate the PCF CI in clinical practice. PMID- 16372696 TI - Ultrasound guidance in intracranial tumor resection: correlation with postoperative magnetic resonance findings. AB - PURPOSE: To determine the inter-method agreement between intraoperative ultrasonography and postoperative contrast-enhanced magnetic resonance imaging (MRI) in detecting tumor residue. MATERIAL AND METHODS: After resection was completed, the cavity borders of 32 tumors were examined with a 7 MHz intraoperative probe. Any echogenic region >5 mm in thickness extending from the surgical cavity into the brain substance was taken as the sonographic criterion for residual tumor. A continuous echogenic rim< 5 mm was considered normal. Results were correlated with gadolinium-enhanced MRI obtained within 48 h after surgery. RESULTS: The kappa value for inter-method agreement was 0.72. There were four cases in whom MRI showed residue despite a negative sonography: extensive edema or Surgicel along the cavity borders (three cases with glioblastoma multiforme) and the cystic component in the vicinity of cerebrospinal fluid (a case with pituitary macroadenoma) may be the reason for the residue going undetected. In a case with glioblastoma multiforme, residual enhancement was < 5 mm in thickness. CONCLUSION: Intraoperative ultrasound is an effective tool for maximizing the extent of intracranial tumor resection. Surgical use has to be minimized if intraoperative ultrasound is to be used as an adjunct to surgery. Tumors with preoperatively detected cystic components in the proximity of CSF containing spaces have to be carefully evaluated with intraoperative ultrasound if residual cystic components are to be detected. A low-thickness echogenic rim should not be considered a reliable sign of the absence of residue. PMID- 16372697 TI - Comparison of adrenal vein sampling value between aldosterone producing adrenal adenoma and non-functioning adrenal adenoma: evaluation using receiver operating characteristic analysis. AB - PURPOSE: To evaluate adequate criteria for adrenal vein sampling values in patients with aldosterone producing adrenal adenoma (APA), retrospectively. MATERIAL AND METHODS: Between 1988 and 2002, 59 hypertensive patients (15 M and 44 F, aged 47.58 +/- 9.45 years) were referred to our hospital and diagnosed with APA based on established criteria. During the same period, 23 patients with non functioning adrenal adenoma (11 M and 12 F aged 53.56 +/- 11.76 years) were diagnosed based on computed tomography and laboratory data. RESULTS: All 82 patients were enrolled in the present study. Bilateral adrenal vein sampling (AVS) for measurement of plasma aldosterone (A) and cortisol (C) was performed, and a receiver operating characteristics (ROC) analysis was conducted to establish the best criteria from the AVS-derived index in patients with APA. A (APA side)/A (contralateral side) was confirmed to provide the best diagnostic accuracy {(>2.5: right APA, sensitivity 83.3%, specificity 79.5%), (> 3: left APA, sensitivity 84.2%, specificity 76.9)}. The Az values for A (APA side)/A (contralateral side) were 0.8948 and 0.9260 for right and left APA, respectively. CONCLUSION: The A (APA side)/A (contralateral side) value was the best compromise for sensitivity and false-positive rate for lateralization of APA. PMID- 16372698 TI - Low-dose and standard-dose unenhanced helical computed tomography for the assessment of acute renal colic: prospective comparative study. AB - PURPOSE: To compare the efficacy of low-dose and standard-dose computed tomography (CT) for the diagnosis of ureteral stones. MATERIAL AND METHODS: Unenhanced helical CT was performed with both a standard dose (260 mAs, pitch 1.5) and a low dose (50 mAs, pitch 1.5) in 121 patients suspected of having acute renal colic. The two studies were prospectively and independently interpreted for the presence and location of ureteral stones, abnormalities unrelated to stone disease, identification of secondary signs, i.e. hydronephrosis and perinephric stranding, and tissue rim sign. The standard-dose CT images were interpreted by one reviewer and the low-dose CT images independently by two reviewers unaware of the standard-dose CT findings. The findings of the standard and low-dose CT scans were compared with the exact McNemar test. Interobserver agreements were assessed with kappa analysis. The effective radiation doses resulting from two different protocols were calculated by means of commercially available software to which the Monte-Carlo phantom model was given. RESULTS: The sensitivity, specificity, and accuracy of standard-dose CT for detecting ureteral stones were 99%, 93%, and 98%, respectively, whereas for the two reviewers the sensitivity of low-dose CT was 93% and 95%, specificity 86%, and accuracy 92% and 94%. We found no significant differences between standard-dose and low-dose CT in the sensitivity and specificity for diagnosing ureter stones (P >0.05 for both). However, the sensitivity of low-dose CT for detection of 19 stones less than or equal to 2 mm in diameter was 79% and 68%, respectively, for the two reviewers. Low-dose CT was comparable to standard-dose CT in visualizing hydronephrosis and the tissue rim sign. Perinephric stranding was far less clear on low-dose CT. Low-dose CT had the same diagnostic performance as standard-dose CT in diagnosing alternative diseases. Interobserver agreement between the two low-dose CT reviewers in the diagnosis of ureter stones and alternative diseases, the identification of secondary signs, and tissue rim sign were high, with kappa values ranging from 0.769 to 0.968. On standard-dose CT scans, the calculated mean effective radiation dose was 7.30 mSv for males and 10.00 mSv for females. On low-dose CT scans, the calculated mean effective radiation dose was 1.40 mSv for males and 1.97 mSv for females. CONCLUSION: Compared with standard scans using 260 mAs, low dose unenhanced helical CT using a reduced tube current of 50 mAs results in a concomitant decrease in the radiation dose of 81%. Although low-dose CT was limited in its ability to depict small-sized calculi less than or equal to 2 mm, it is still comparable to standard-dose CT for the diagnosis of ureter stones and alternative disease. PMID- 16372699 TI - Computed tomography of the urinary tract: optimalization of low-dose stone protocol in a clinical setting. AB - PURPOSE: To establish whether information would be lost if slice reconstruction thickness was increased from 3 to 5 mm, and whether this altered how difficult it was to interpret the examinations. MATERIAL AND METHODS: Twenty-three consecutive patients referred with suspected or known urinary stones were included. All examinations were performed without intravenous contrast media. The original series, with effective mAs 50, were reconstructed with slice thickness 3 and 5 mm, respectively. All demographic and examination data were removed and the series reviewed in PACS by two independent radiologists. Objective findings, i.e. number and size of stones, signs of obstruction, and evaluation of interpretation difficulty, were registered. RESULTS: Identical findings were registered in 18 of the series of 3 mm (n=23) and 19 of the series of 5 mm (n=23). In two series reconstructed with 3 mm slice thickness and in one series with 5 mm slice thickness, the observers disagreed on the presence of urinary stones. Main reasons for interpretation difficulties were given as "lack of intra-abdominal fat" and "many phleboliths in the pelvic region", but never "disturbing noise". CONCLUSION: To determine the presence and size of urinary stones at low-dose computed tomography, 5 mm reconstruction algorithm seems equal to 3 mm. Patient related factors influence the interpretation more than image quality. PMID- 16372700 TI - Polyorchidism: sonographic and magnetic resonance image findings. AB - Polyorchidism is a rare congenital anomaly frequently associated with maldescent testis, hernia, and torsion. Reports in the literature show an increased risk of testicular malignancy in the presence of polyorchidism. This entity has characteristic sonographic features and the diagnosis is often made on the basis of sonography. Magnetic resonance imaging might also be used for the diagnosis, but is more helpful in cases associated with cryptorchism or neoplasia. A conservative approach is the treatment of choice in uncomplicated cases. PMID- 16372702 TI - The evidence is there--let's use it. PMID- 16372703 TI - Parents' management of their child's postoperative pain. AB - In spite of its benefits, day or minor surgery is not without complications; pain is a significant issue for someatients (Kotiniemi et al 1997, Grenier et al 1998). Having witnessed considerable variation in the discharge advice given to parents about pain control we were prompted to discover what evidence there is to suggest 4 hat parents can effectively manage their child's postoperative pain at home following day or minor surgery. In this artide cle review the evidence and describe how review findings were used to inform practice. PMID- 16372704 TI - Dependency scoring for child and adolescent inpatient mental health services. AB - Child and adolescent mental health services (CAMHS) inpatient units provide multi disciplinary treatment for young people with a wide range of complex needs, many of whom have co-morbid presentations. There is no reliable, validated measure of CAMHS patient dependency. This article reports the first stages of a project to develop a properly researched dependency measure for use before (i.e. by those referring the young person) and during the young person's stay. Expert opinion was sought to develop a list of statements regarding a young person's care which could be rated according to level of need. Initial piloting demonstrated face and concurrent validity; next steps include exploring issues of item ambiguity and weighting. PMID- 16372705 TI - Noise levels in PICU: an evaluative study. AB - High levels of noise in the hospital environment can have an impact on patients and staff increasing both recovery time and stress respectively. When our seven bedded paediatric intensive care unit (PICU) is full, noise levels seem to increase significantly. This study measured noise levels at various times and places within a PICU using Tenma sound level meter which simulates the subjective response of a human ear. Noise levels were often excessive, exceeding international guidelines. Staff conversation was responsible for most of the noise produced; medical equipment, patient interventions, telephones, doorbell and the air shoot system were also responsible for causing high levels of noise. More can be done to reduce noise and its effects on patients and staff. PMID- 16372706 TI - The bully as victim? AB - Bullying in schools is now widely acknowledged and the detrimental effect on children who are bullied is well recognised. A literature review aimed at exploring the links between bullying and depression in primary school children uncovered evidence of depression in the children who bully. Further analysis of the literature identified conflicting research findings. This article summarises the causes and implications of aggression and bullying behaviour, including future outcomes for children who bully. Further research is necessary as the evidence suggesting that bullies are depressed contradicts indications that they are confident and have a high self-esteem. PMID- 16372707 TI - Multicentre research: lessons from the field. AB - Multicentre research can be used to explore and generate significant data in aspects of care that affect small numbers of children. This article describes the problems and benefits encountered by a group of nurses from 11 hospitals undertaking a multicentre study of pressure ulcers in children and young people in England and Wales. Multicentre research can generate a large amount of useful data contributing to high quality evidence-based care and can provide nurses with a valuable learning and networking experience. PMID- 16372708 TI - The Integrated Children's System: challenges of information sharing. AB - Information has to be shared within and between agencies if Every Child Matters (DH 2003a) is to bring real benefits to children in need and their families. The Integrated Children's System is a theoretical framework that offers a single approach to assessing, planning, intervention and reviewing based on an understanding of children's developmental needs in the context of their families and communities. It should not be seen as an IT project, but is closely linked to the electronic social care record and to NHS care record service developments. Health and social care professionals attempting to improve information management as part of better interagency working face a number of challenges including: confidentiality; a single identifier for children; common language and information exchange; system, organisational and staff issues. Despite many benefits, implementation of the integrated children's system presents managerial and organisational challenges which will require additional support. PMID- 16372709 TI - Living with disability: part 1. AB - This is the first of three articles which consider the holistic care required by children with a disability and their families. Although the articles mainly address care of children with disabilities in the acute hospital setting, the concepts and knowledge can be transferred to care in other settings. The articles aim to provide a summary of the main subject areas while prompting readers to revisit their own opinions and experiences. This article provides an overview of: commonly used terminolgy; receiving the news that your child has a disability; impact on family members; and relevant social policy. The second and third articles cover: the therapeutic use of self; the value of communication; meeting complex needs on a paediatric ward and multi-diciplinary working. PMID- 16372710 TI - Evaluation: part II: evaluating a provider unit. PMID- 16372711 TI - Melanoma. PMID- 16372712 TI - Teaching for higher order learning. PMID- 16372713 TI - Nursing competency assessment across the continuum of care. AB - Establishing a thorough and effective competency assessment program is essential to meeting standards of regulatory bodies and providing quality care. Nursing education specialists continue to strive to provide a competency assessment process that is meaningful and uncomplicated. This article describes the implementation of a nursing competency assessment program in a large midwestern medical center that spans the continuum of care and incorporates all nursing roles that require nursing licensure, including advanced practice nurses, into one centralized nursing competency program. Competency program infrastructure, topic identification, documentation to support the evaluation and validation process, and continuous improvement opportunities are described. PMID- 16372714 TI - Models, measurement, and strategies in developing critical-thinking skills. AB - Health care professionals must use critical-thinking skills to solve increasingly complex problems. Educators need to help nurses develop their critical-thinking skills to maintain and enhance their competence. This article reviews various models of critical thinking, as well as methods used to evaluate critical thinking. Specific educational strategies to develop nurses' critical-thinking skills are discussed. Additional research studies are needed to determine how the process of nursing practice can nurture and develop critical-thinking skills, and which strategies are most effective in developing and evaluating critical thinking. PMID- 16372715 TI - Continuing education needs assessment of acute care and long-term-care nurses in a Veterans Affairs Medical Center. AB - BACKGROUND: This study was done to identify nurses' priorities for continued learning and to examine the priorities in relation to age, educational level, location in the organization, experience, position in the organization, and shift worked. METHODS: A random sample of nurses at a Veterans Affairs Medical Center completed surveys consisting of 58 educational topics and demographic information. The results were analyzed using descriptive statistics, analysis of variance, and Tukey's HSD method to identify differences within the various subgroups of nurses. RESULTS: Of the 58 educational topics, 21 were determined to be high priority educational needs and were significantly correlated with one or more of the independent variables. CONCLUSION: Conducting a needs assessment can provide valuable information that will improve the delivery of high quality, relevant continuing education and improve staff satisfaction. PMID- 16372716 TI - Nursing education and service collaboration: making a difference in the clinical learning environment. AB - This article focuses on innovative collaborative steps that were identified in recent research conducted by these authors on the relationship between academia and service. These steps are currently being implemented in the hope of improving the important role that the clinical environment plays in student nurses' education. Few factors in nursing education are as important as the clinical environment in which students do their training. This article elaborates on these steps and offers practical suggestions for improving the relationship between academia and service. PMID- 16372718 TI - The coming drug bust? PMID- 16372717 TI - Patient care delivery model improves nurse job satisfaction. AB - BACKGROUND: Nurse job satisfaction was identified as an outcome measure for a project to develop and implement a patient care delivery model in an acute psychiatric setting. METHOD: Seventy registered nurses were surveyed during 3 years, utilizing the Index of Work Satisfaction. RESULTS: Nurse job satisfaction has improved significantly (14%) since model development began. Analysis of variance showed significant (p < .05) increases in all component mean scores in the second and third surveys. CONCLUSION: The relationship-based nursing model empowers nurses who know the patient best to decide how to provide care. It affirms the values that are the foundation of nursing practice, creating an environment where nurses feel they make a difference. PMID- 16372719 TI - Sex, drugs and hope. PMID- 16372720 TI - Who gets the organs? PMID- 16372721 TI - Charles Darwin: evolution of a scientist. PMID- 16372722 TI - The things they carry. PMID- 16372723 TI - What caused the flu vaccine crisis? PMID- 16372724 TI - Alcohol compromises mental health care. PMID- 16372725 TI - A nursing dynasty. PMID- 16372726 TI - Liberating patients with HIV. PMID- 16372727 TI - How to improve the safety of patients treated in the NHS. AB - The NHS relies on a variety of different staff, medicines and equipment in order to provide health care. Coupled with the volume of patients being treated, this can make it difficult to maintain safety. A new report from the National Audit Office looks at what can be done to make the NHS a safer environment for patients. PMID- 16372728 TI - Scleroderma. PMID- 16372729 TI - The liver. Part 2: Physiology. PMID- 16372730 TI - Detecting hyperprolactinaemia in mental health patients. AB - Hyperprolactinaemia is associated with drugs used in mental health to treat psychosis (antipsychotics) and affects a significant number of service users. The condition causes endocrine disturbances, including sexual dysfunction, amenorrhoea and infertility. In the longer term, service users are also at risk of disabling conditions such as osteoporosis. Early detection of hyperprolactinaemia enables safer use of antipsychotic drugs. This article focuses on neuroendocrine adverse effects and how this problem can be addressed. PMID- 16372731 TI - Implementing a patient-led service for chronic conditions. AB - Many chronic conditions with fluctuating levels of disease activity are traditionally managed by lifelong regular medical reviews. However, this means appointments do not always coincide with patient need, while the volume of reviews makes it difficult to respond quickly to requests for help. Research in rheumatoid arthritis suggests that hospital-initiated reviews can be replaced by patient-initiated reviews, supported by nurse-led initiatives. PMID- 16372732 TI - Clopidogrel. PMID- 16372733 TI - Importance of early referral in sudden loss of hearing. AB - AIM: To examine the personal and clinical characteristics of patients suffering from sudden hearing loss. METHOD: The participants were 45 Israeli patients, and the data was collected retrospectively from their medical files. RESULTS: In most cases the injury was sensory, and in one ear. The common symptom was tinnitus, followed by vertigo. In 20 per cent of the cases no preliminary signs were documented. As many as 60 per cent of the participants suffered from high levels of cholesterol. This finding might indicate a link between the events of hearing loss and patients' vascular background. A total of 42 per cent of the participants delayed their appeal to treatment, despite the appearance of symptoms. CONCLUSION: A significant link was found between quick referral to treatment and recovery. The nurse holds a central role in identifying this group of patients and ensuring their referral to further treatment. PMID- 16372734 TI - How to excel at giving appraisals. PMID- 16372735 TI - Extending the boundaries of care. PMID- 16372736 TI - Rear view. In this modern age it is possible for us to have knowledge and not feel obliged to do something. PMID- 16372738 TI - Nothing to smile about. A cautionary tale for dentists about electronic communication. AB - We've all heard the adage "good news travels, but bad news travels faster." In the age of the Internet, bad news has the ability to reach millions of people in a matter or minutes. While many dentists know how to address disgruntled patients, most will never have to experience a full-fledged crisis as a result of one patient's bad experience. Dr. Rodney Shivers of Oklahoma City was the victim of a spurious e-mail smear campaign that led to his receiving thousands of harassing phone calls, threatening letters and media attention. By sharing his experience in addressing this situation, he hopes to help other dentists avoid his fate. PMID- 16372739 TI - Small business owner: retirement plans. PMID- 16372740 TI - The atrophic mandible. PMID- 16372741 TI - Medicare drug plan may not be living up to savings promises. PMID- 16372742 TI - 10 initiatives may ensure uninterrupted drug coverage for dual eligibles. PMID- 16372743 TI - Budget reconciliation proposals shift Medicaid costs to states, NGA says. PMID- 16372744 TI - Did nurses have duty to tell pt. what happened? PMID- 16372745 TI - 'Unlicensed nurse' fails to consult Dr.: quadraplegic infant. PMID- 16372746 TI - MN:LPN fired for yelling obscenities at pt., etc.: unemployment benefits denied- firing for cause. PMID- 16372747 TI - Nurse terminated for abusive conduct toward mental patient. PMID- 16372748 TI - [Hematuria and acanthocyturia in patients with diabetes]. AB - It has been suggested that the finding of acanthocyturia in patients with hematuria points to the presence of glomerulonephritis. However, little attention has been paid to the finding of acanthocyturia in diabetic patients with hematuria. Here we studied 93 consecutive diabetic patients and explored the prevalence of microscopic hematuria and acanthocyturia in association with normoalbuminuria (NO; urinary albumin excretion (UAE) of < 30 mg/g x creatinine), microalbuminuria (MI; UAE of 30-299mg/g x creatinine), macroalbuminuriaMA; UAE of > or = 300mg/g x creatinine), or chronic renal failure (CRF; serum creatinine levels of > or = 1.1mg/dl for male and > or = 0.9mg/dl for female). We defined microscopic hematuria as > or = 5 erythrocytes per high-power field and acanthocyturia as > or = 5% acanthocytes (erythrocytes of doughnut-like appearance with vesicle-shaped protrusions) among 100 erythrocytes in the centrifuged urinary sediment. Microscopic hematuria was found in 12 (24%) out of 49 patients with NO, in 9 (43%) out of 21 patients with MI, in 6 (75%) out of 8 patients with MA, and in 7 (47%) out of 15 patients with CRF. Patients with microscopic hematuria showed a significant increase in urinary albumin excretion as compared to those without (836 +/- 265 vs. 135 +/- 56, p < 0.01). Of patients with microscopic hematuria, acanthocyturia was observed only in 2 (22%) out of 9 patients with MI and in 2 (33%) out of 6 patients with MA. Two of 4 patients with acanthocyturia had elevated serum levels of IgA and chronic tonsillitis, which indicated the occurrence of IgA nephropathy in these patients. Thus, microscopic hematuria was common and associated with elevated UAE, while acanthocyturia was rare and observed only in patients with elevated UAE. We propose that more attention should be paid to the finding of acanthocyturia in diabetic patients with hematuria and albuminuria. PMID- 16372749 TI - The usefulness of immunostaining in washed cell cytological smears for differential diagnosis of B cell type malignant lymphoma. AB - We investigated the usefulness of immunostaining on washed cell cytological smears for the differential diagnosis of B-cell type malignant lymphoma. Twenty eight cases with possible malignant lymphoma were examined. The tissues were squashed in a test tube of isotonic saline. The cell suspensions were then washed to remove non-specific soluble immmunoglobulin, and were smeared. Immunocytochemical staining for IgG, IgA, IgM, kappa and lambda was performed on the smears. As seen in the results, all cases of B cell lymphoma have a positive rate of 20% or over for kappa or lambda, and a kappa/lambda ratio of over 4.5 or a ratio of over 2.0 lambda/kappa, in the washed cell smears. Thus, it is possible to distinguish B cell lymphoma from the others by means of the results use as cut off scores. In addition, the rapid staining protocol in the present study was able to provide the results within 15 min. Therefore, it might be the most suitable method for the detection of immunoglobulin monoclonality in general laboratories. PMID- 16372750 TI - [Alcohol dehydrogenase interference in clinical chemical laboratory tests]. AB - Alcohol dehydrogenase (AD, EC 1.1.1.1 ) activity in serum markedly increases in cases of hypoxic hepatic injury and acute hepatitis. Furthermore, AD can reduce oxidized coenzyme NAD+ to NADH even without substrate such as ethanol. Therefore, AD interferes in clinical chemical tests using the reducing reaction of coenzyme NAD+. We found that AD caused a false positive in serum glucose determination using an enzymatic procedure (hexokinase/glucose-6-phosphate dehydrogenase/NAD+ coupled assay) and increased the blank value of the sample on lactate dehydrogenase (LD) assay using the method recommended method by JSCC (Japanese Society of Clinical Chemistry). We should approve the correction of sample blank reaction in LD activity assay using the method recommended by JSCC as well as the IFCC (International Federation of Clinical Chemistry) method. Also, we should adopt the enzymatic procedures using the reducing of coenzyme NADP+ or without the influence of AD. PMID- 16372751 TI - [Detection of antibodies specific for blood group antigens A and B]. AB - The measurement of anti-blood group A/B (anti-A/B) IgG antibody levels is important for ABO unmatched-organ recipients because the effective removal of the antibodies improves their prognosis. Living-donor liver transplantation (LDLT) into ABO-unmatched patients tends to have a very poor outcome due to major complications such as intrahepatic bile duct complications and hepatic necrosis. Sustained bile duct complications are associated with high preoperative IgM type anti-A/B Ab titers, while patients with high preoperative IgG type anti-A/B Ab titers frequently develop sustained hepatic necrosis. There are several existing methods by which anti-A/B Ab levels can be measured, including the standard tube (TT) method, an enzyme-linked immunosorbent assay, and a flow cytometry method. Anti-A/B IgG Ab is difficult to identify by the TT method, which is the most popular method and is based on the detection of hemagglutination, because the major isotype that facilitates red cell agglutination is the pentameric IgM molecule. Therefore, we have developed a method based on surface plasmon resonance (SPR) that detects the presence of the antigen-antibody complex without any labeling. This method allows us to rapidly quantitate anti-A/B IgG Ab levels. PMID- 16372752 TI - [Trend on medical economy and its relation to laboratory testing]. PMID- 16372753 TI - [Progress in assays for and new developments in autoimmune pathophysioirmen's introductory remarks]. AB - We examined the influence of a revision of health insurance in 2004 in Teikyo University Hospital in comparison with 2003. Out to the returns of most hematological tests, especially HbA1c and TK, being included in the treatment charges of outpatients, gross laboratory test sales decreased more than expected. Increased income from judgment charges and management charges could not make up for the deficit in test sales. Therefore, total laboratory test revenues decreased substantially. In 2006, the new revision of health insurance will worsen the revenues further. Hospital laboratory test departments should take appropriate and aggressive management steps in response to these revisions in health insurance. PMID- 16372754 TI - [Activity of clinical laboratory physicians in national health insurance]. AB - As places of activity of clinical laboratory physicians in national health insurance, there are committees of the Japan Medical Association and the Social Insurance Union of Societies Related to Internal Medicine. In 1996, additional management fee was approved due to the efforts of the then President Kinya Kono, and subsequently, fees were increased after every revision, markedly contributing to the income at the department of clinical laboratory in hospitals. Another activity of clinical laboratory physicians is work in the Social Insurance Union of Societies Related to Internal Medicine and that Related to Surgery toward the 2006 revision of medical insurance fees. The former union presented a request paper in cooperation with each scientific society for the revision of medical insurance fees, and clinical laboratory physicians played the central role in summarizing requests associated with laboratory tests. We describe the contents of the request paper for the 2006 revision of medical insurance fees presented by this union. In the previous revisions (2004), after the Ministry of Health, Labour, and Welfare counseled with the Japan Medical Association, the Central Social Insurance Medical Council made the final decisions. In the future, the power of the Ministry of Health Labour, and Welfare may increase. In activities of clinical laboratory physicians in medical insurance fees, clinical laboratory tests and physiological function tests should be evaluated according to each items, and the income/expenditure balance should be improved. In the 2006 revision of medical insurance fees, a marked reduction in medical costs for the increase in medical insurance for the elderly is expected. For this reduction, laboratory tests for which a reduction is easy may become targets. The government reported the principle that reimbursement in 2006 will be reduced by 2-5% to put the brakes on the rise in national medical costs due to the aging of society. The prospective payment system mainly by the diagnosis procedure combination (DPC) is performed mainly at university hospitals at present, and attention is paid to its future progress. PMID- 16372755 TI - [Medical technologist as a member of infection control team]. AB - For the prevention of infection at institutions, an Anti-nosocomial Infection Committee or an Infection Control Team (ICT) is organized at each institution according to its scale. We report the present status of the ICT managed mainly by medical technologists engaged in microbiological examination (certified medical microbiological technologists) at Dokkyo University School of Medicine. Since this hospital is an educational hospital, the department of clinical laboratory medicine cooperates with the microbiological laboratory of the clinical laboratory in infection control education of medical workers (such as medical students, nursing students, physicians and nurses) in infection diagnosis, infection control/infection management. Since infection control is achieved by improvement in hygiene knowledge and its practice in all citizens, we also attached importance to publicity activities associated with microbiology for patients, their families, and all medical workers. PMID- 16372756 TI - [Participation of medical technologists in the nutrition support team (NST)]. AB - In the nutrition support team (NST), medical technologists take a very important role in carrying out nutrition screening, nutrition assessment and nutrition monitoring. Medical technologists can look at the laboratory data of patients earlier than other NST staff. Using this advantage, the medical technologists can contribute to the NST. In addition, medical technologists join ward rounds together with other NST staff on a weekly basis. After discussion with all NST staff, medical technologists make a report on the nutrition assessment of patients and then sign their names to the report. The NST is a representative of medical treatment teams. All NST members should have equal levels of understanding of nutrition therapy. Therefore, medical technologists should not only look at the laboratory data of the patients, but also manage the patients through nutrition assessment and nutrition monitoring. PMID- 16372757 TI - [Participation of medical technologists in educational hospitalization of diabetics--involvement in short-term hospitalization of diabetics on weekends]. AB - Since diabetic mellitus often causes few symptoms, the evaluation and control of the condition by clinical examinations are important. Patients require long-term self-control, and therefore, should understand the importance of various examinations, appropriate ways of undergoing examinations, and their own physical condition based on examination values for the continuation of good control. However, it is often difficult to provide adequate information, give instructions, evaluate the degree of understanding by patients, or confirm changes in their detailed feelings within the limited consultation time at the outpatient clinic. One of the purposes of educational hospitalization of diabetics is to overcome these problems. To make good use of educational hospitalization of diabetics, staff members concerned should establish an effective cooperation system based on common information and recognition, and give appropriate instructions. Clinical technologists evaluate samples in their work and are the first to know measurement values. Therefore, it may be of great value in the establishment/continuation of good diabetic care that medical technologists well-informed about examination work become involved in educational hospitalization of diabetics, understand the condition of patients and requests by staff members in other departments, and rapidly provide information. In addition, consideration should be given to the educational hospitalization period so as to minimize patient's burden. PMID- 16372758 TI - [Participation of medical technologists in the liver disease seminar]. AB - Since September 2002, the Koshigaya Hospital Dokkyo University School of Medicine (KHDUSM) has been holding a liver disease seminar every three months. The members are consisted of gastroenterologists and co-medicals (nurses, pharmacists, nutritionists, and medical technologists) from various departments. Aims of this seminar offer the accurate medical information of liver disease to help the patients to understand their diseases, facilitating their active involvement in the treatment. Technical and easy-to-understand descriptions by the co-medicals are necessary to improve the QOL of patients. The seminar has been promoting the collaboration and the cooperation between physicians and co-medicals with the relationship to patients. I hope that the seminar can further contribute to the optimal treatment of liver diseases. This paper introduces the liver disease seminars at KHDUSM, from its preparation to the opening, especially by the clinical laboratory department. PMID- 16372759 TI - [Diet counseling through "Shoku-dietary Coaching"]. AB - "Shoku-dietary Coaching" is a skill under development by Kageyama, who applies "coaching," widely used in the business field, to diet counseling. This counseling aims at improving conventional "nutritional guidance-type diet counseling" and promoting self-motivation so that healthy clients eagerly improve their own health, and clients with obesity or lifestyle-related diseases can learn self-control. In Shoku-dietary Coaching, the basis for the differentiation between healthy and unhealthy conditions is not only the parameters measured by medical devices. In Shoku-dietary Coaching, attention is directed to clients' assessment of their own lifestyle, dietary goals they have, and actions they will take to achieve them. To increase the health level of clients, we are developing techniques to enhance their motivation by showing sympathy with and support for their dietary behavior and health awareness. In addition, we give guidance through both theory and the practice of such things as having three meals a day at regular hours, knowing the kinds and daily amounts of foods appropriate for each client, and clarifying the percentages of seasonings necessary for cooking. The habit of having meals at regular hours alleviates stress, promotes communication with people sitting at the same table, and increases the health level of both the client and the others. These are important elements in the theory of Shoku-dietary Coaching. Putting the above into practice should not be limited to clients, but should include the clinic staff so as to deepen their own understanding and communication. Enhanced communication reinforces team medical care in the clinic. Communication skills which involve respect for others, continuous motivation of individuals, and achievement of purposes that may even require a long time may be useful for all people. PMID- 16372760 TI - [Studies on gene and structure and function of Clostridium botulinum toxin]. PMID- 16372761 TI - [New paradigm of membrane transport: ER-to-Golgi trafficking of the lipid ceramide by a molecular extraction and transfer mechanism]. PMID- 16372762 TI - [Protein crystal structures for analysis of protein functions and drug design]. PMID- 16372763 TI - [Topics in Quorum-sensing research: from inter-species, inter-kingdom communication to recognition of host responses by bacteria]. PMID- 16372764 TI - [Exploration of the breeding ground for increasing medical errors]. PMID- 16372765 TI - [Diagnosis for GERD--up to date]. PMID- 16372766 TI - [Endoscopic treatment for gastroesophageal reflux disease]. PMID- 16372767 TI - [A case of chronic hepatitis C complicating acute cholecystitis with blood clots in the gallbladder due to hemobilia after percutaneous liver biopsy]. PMID- 16372768 TI - [Gastrointestinal stromal tumor successfully treated with re-administration of STI571 despite of serious skin rash: a case report]. PMID- 16372769 TI - [A case of AFP-producing gastric cancer with multiple liver metastasis responding to TS-1 and cisplatin combination chemotherapy]. PMID- 16372770 TI - [A case of focal fatty liver occurring after gastrectomy with difficulty in distinction from metastatic liver cancer ]. PMID- 16372771 TI - [A case of phlegmonous esophagogastritis improved with conservative therapy]. PMID- 16372772 TI - [Case report: primary adenocarcinoma of the thoracic esophagus that probably originated in the esophageal gland]. PMID- 16372775 TI - Agenda for chaos. PMID- 16372776 TI - Hospital acquired infections what's going wrong? PMID- 16372778 TI - Dealing with the media. PMID- 16372777 TI - Razors versus clippers. A randomised controlled trial. AB - The purpose of this randomised controlled trial was to determine if patients showed a preference for preoperative hair removal with razors or clippers and to identify if one method was associated with more trauma or postoperative infections. The trial took place in a day surgery unit with patients who were having a range of surgical procedures including hernias and varicose veins. This study was sponsored by an award from the NATN/3M Clinical Fellowship. PMID- 16372779 TI - Walk or be driven? A study on walking patients to the operating theatre. AB - The objective of this study was to determine patients' perception on the possibility of changing a tradition-based practice to a more patient-empowering service, thus demonstrating that nurses can make a difference in providing a more patient-centred environment. The study revealed that the patients themselves wanted to be included in the decision-making process and actively embrace change. PMID- 16372780 TI - Waiting lists. What are we doing right? AB - According to The Sunday Times Good Hospital Guide (2004), the Mater Hospital scored highest for waiting lists, admitting 84% of inpatients within six months of referral. To achieve this result the Trust was originally invited by commissioners to submit a bid to develop a service which ensures that patients currently waiting for elective surgery receive it as quickly as possible. PMID- 16372781 TI - Waiting lists? What waiting lists? Not nursing's problem. PMID- 16372783 TI - Judith Shamian: be true to yourself and your values. PMID- 16372784 TI - The legacy of leaders. PMID- 16372785 TI - Health human resources planning in an interdisciplinary care environment: to dream the impossible dream? PMID- 16372786 TI - Enhancing skills for evidence-based healthcare leadership: the Executive Training for Research Application (EXTRA) program. AB - The Executive Training for Research Application (EXTRA) is a new training program that aims to increase the skills of health services executives and their organizations to use research evidence in healthcare management and decision making. This paper describes the goals and rationale of the EXTRA program and its learning objectives and curriculum, and reports on some early baseline evaluative research. In particular, the authors address the opportunities that EXTRA offers to leaders in the nursing profession to transform the practice of nursing and patient care, and the unique opportunities that the program offers for collaboration across the healthcare professions and disciplines. While the EXTRA training program requires substantive investment of time and commitment by healthcare leaders and their organizations, it offers great potential for increasing research application in healthcare leadership decision-making. It is therefore a potential long-term lever of cultural decision-making change within healthcare organizations. PMID- 16372788 TI - Are skills learned in nursing transferable to other careers? AB - OBJECTIVE: To determine the influence of skills gained in nursing on the transition to a non-nursing career. BACKGROUND: Little is known about the impact that nursing skills have on the transition to new careers or about the transferability of nursing skills to professions outside nursing. METHOD: A postal questionnaire was mailed to respondents who had left nursing. The questionnaire included demographic, nursing education and practice information, reasons for entering and leaving nursing, perceptions of the skills gained in nursing and the ease of adjustment to a new career. Data analysis included exploratory and confirmatory factor analysis, Pearson product moment correlations and linear and multiple regression analysis. RESULTS: Skills learned as a nurse that were valuable in acquiring a career outside nursing formed two factors, including "management of self and others" and "knowledge and skills learned," explaining 32% of the variation. The highest educational achievement while working as a nurse, choosing nursing as a "default choice," leaving nursing because of "worklife/homelife balance" and the skills of "management of self and others" and "knowledge and skills" had a significant relationship with difficulty adjusting to a non-nursing work role and, overall, explained 28% of the variation in this difficulty adjusting. CONCLUSION: General knowledge and skills learned in nursing prove beneficial in adjusting to roles outside nursing. PMID- 16372787 TI - Nurses' use of research in practice at three US Army hospitals. AB - Translation of research into clinical nursing practice has been extensively studied with nurses in civilian institutions. However, limited examination has been made of research utilization in US military facilities. A quantitative survey approach, using Estabrooks's adapted Research Utilization Survey and Mylle's Organizational Climate Survey, was employed to determine the extent that registered nurses in selected US Army Medical Treatment Facilities (MTFs) use research findings for their own practice and to describe the factors, both professional and organizational, that enhance or hinder research utilization. This study found that nurses at the MTFs reported using research in their practice less than half the time. Variables most significantly correlated with research utilization were belief suspension and attitudes toward research. The variable of support correlated significantly with all types of research utilization except indirect. Other organizational factors influencing nurses' use of research findings in practice were time, accessibility to research and a champion to assist their efforts. Interestingly, the number of years worked in nursing and in the facilities correlated inversely with research utilization, suggesting that over time, nurses may begin to rely on past experience rather than seek out new knowledge for practice. PMID- 16372789 TI - A span of control tool for clinical managers. AB - During the second half of the 1990s, healthcare in Canada experienced significant downsizing and reform. One of the consequences of these reorganizations has been a reduction in the number of clinical managers and a significant increase in their span of control, to the point that often their abilities to fulfil their role as clinical managers are hindered (Altaffer 1998; Counsell et al. 2001; Pabst 1993). The first-line manager plays a critical role in the delivery of healthcare, in particular, within nursing services. Therefore, providing support for the professional practice of clinical managers should become a priority. PMID- 16372790 TI - Hippocratic oath in the modern times. PMID- 16372791 TI - Cytomorphometric analysis of the buccal mucosa of tobacco users. AB - OBJECTIVE: This study has been carried out to assess the effect of tobacco smoking and of betel quid chewing with tobacco on buccal mucosa by cytomorphometry, in a south Indian population. STUDY DESIGN: Cellular diameter (CD) and nuclear diameter (ND) of exfoliated buccal squames obtained from clinically normal appearing buccal mucosa of tobacco smokers, betel quid with tobacco chewers, and those with a combined habit, stained by the Papanicolaou method, were measured. Non-users served as negative controls and oral squamous cell carcinomas in tobacco users served as positive controls. One way ANOVA test of the values obtained followed by multiple range comparison with Tukey-HSD procedure (at p=0.05) was carried out. RESULTS: A statistically significant reduction in CD and increase in ND in smokers and those with a combined habit were observed. CONCLUSION: The use of tobacco in the form of smoking influences the cytomorphology of buccal mucosa. PMID- 16372792 TI - Comparative efficacy of aceclofenac and etoricoxib in post extraction pain control: randomized control trial. AB - The study is a prospective randomized double blind clinical trial comparing the efficacy of nonselective NSAIDs (Aceclofenac) and highly selective COX-2 inhibitors (Etoricoxib) in post extraction pain control. The primary efficacy was judged by overall assessment of onset and duration of analgesic effect and rate of decrease in pain intensity by a visual analogue scale over a 3-day investigation period. 100 patients were enrolled in the study (50 patients in aceclofenac group and 50 patients in etoricoxib group). Twice-daily dosage of aceclofenac 100 mg and etoricoxib 60 mg were recommended for the double blind study. 64 patients completed the study. Efficacy of pain control over baseline data documented in both the treatment groups were statistically significant (p<0.05). There was no significant difference between the two drugs. PMID- 16372793 TI - Palatal rugae pattern in Mysorean and Tibetan populations. AB - OBJECTIVES: To compare the palatine rugae pattern of Mysorean and Tibetan subjects, to assess the predominant palatine rugae pattern in Mysorean and Tibetan population, and to determine the uniqueness of palatine rugae pattern in an individual. MATERIALS: Maxillary study models of 60 subjects each (30 males and 30 females) of age group 17-23 years, of Mysorean population and Tibetan population were assessed for the length, shape, and unification of rugae based on the classification by Lysell et al. The palatine rugae were compared for the above mentioned parameters. 2 way ANOVA and Chi square tests were used for statistical analysis. RESULTS: Statistical analysis showed that males had significantly more number of total rugae on the right compared to the left side in both populations. Indian males had more primary rugae on the left side as compared to females and vice versa for the Tibetan population. Also, Indian males had more number of curved rugae on both right and left sides than Tibetan males and Tibetan females had more wavy rugae on right and left sides than Indian females. In other aspects, no significant difference or association was found. CONCLUSIONS: A statistically significant association between the total number of rugae and the gender in both the study populations exists. Parameters like the length and shape of the rugae show racial differences. PMID- 16372794 TI - Effects of elevated temperatures on various restorative materials: an in vitro study. AB - In cases of mass disasters associated with fire, identification of the burnt victims can be a real challenge to the forensic team. Teeth and their restorations play a significant role to aid in the identification process, as various restorative materials have varying resistance to high temperatures. A study was undertaken to evaluate the changes taking place on teeth restored with amalgam, composites, glass ionomers, heat cure acrylic, and ceramics. The specimens were placed in a furnace and heated to predetermined temperatures of 200, 400, 600, 800, and 1000 degrees C and the changes were examined using a digital camera and stereomicroscope. Our observations show that while some restorations were able to withstand elevated temperatures, others were reduced to an unrecognizable mass at relatively low temperatures. PMID- 16372795 TI - Toothbrush age, wear, and plaque control. AB - OBJECTIVE: This study aims to assess the effect of toothbrush age and wear on plaque control. MATERIALS AND METHODS: 36 dental students were recruited for the 3-month double blind trail. Each participant received commercially purchased soft toothbrush and nonfluoridated toothpaste. Group I (n= 18) used the same brush for 3 months and Group II (n=18) replaced the brush every month. Toothbrush wear was recorded using Rawls formula (1989) and Plaque score was recorded using Patient Hygiene Performance index (1968). Recalls for the participants were scheduled after 1 (T1), 2 (T2) and 3 months (T3). OBSERVATIONS: Results indicated that the toothbrush wear varied widely amongst the participants. 25-62% wear was observed in Group I and 6-24% in Group II. The participants using the same toothbrush for 3-months did not exhibit statistically significant different plaque scores compared to those who replaced their toothbrush every month. Toothbrush age and wear was not related to plaque control. PMID- 16372796 TI - Treatment of gingival recession by modified Langer and Langer technique. AB - In recent years, stronger need for esthetics and growing cosmetic demand for a pleasing smile has made many individuals more conscious of the recession on facial aspect of the teeth. Here we report a case illustrating treatment for facial gingival recession using modified Langer and Langer technique. The clinical results showed good soft tissue coverage of the root surfaces and improving esthetics. PMID- 16372797 TI - Bilateral mandibular canines with type two canals. AB - Presence of two canals with type II pattern in mandibular canine is around 15%. Even though the incidence rate is high, multiple angled diagnostic radiographs are not regularly taken. This article highlights the importance of multiple angled radiographs and indicates that the dentist should have a mind set to look out for extra canals in all root canal cases. PMID- 16372798 TI - [Skin and teeth]. PMID- 16372799 TI - Pyoderma gangrenosum. AB - Pyoderma gangrenosum (PG) is a non-infectious reactive neutrophilic dermatosis which typically starts with pustules which rapidly evolve to painful ulcers of variable size and depth with undermined violaceous borders. Since its first description in 1930, the pathogenesis of PG has remained elusive even as an ever widening range of systemic diseases has been described in association with it. The diagnosis of PG is based on clinical and pathologic features and requires exclusion of other conditions that produce ulcerations, since misdiagnosis exposes patients to risks associated with treatment. Critical to proper management are correct diagnosis, identification and treatment of any underlying disorder, and the appropriate choice of topical and systemic therapy. PG has four distinctive clinical and histologic variants, and the specific clinical features of the lesion may provide a clue to the associated disease. The most common associated diseases are inflammatory bowel disease, rheumatological or hematological disease or malignancy. Although there is no single successful treatment for PG, certain type of PG lesions are recognized to respond more readily to accepted therapies than others. Local treatment may be sufficient for mild disease, while systemic immunosuppressive therapy is necessary for severe cases. The treatments with the best clinical evidence are oral or pulse intravenous corticosteroids, and cyclosporine. Surgical therapy is useful in selected cases in conjunction with immunosuppression. Wound stabilization is obtained only through control of the systemic and local inflammatory process. Emerging therapies include use of platelet-derived growth factor and cell culture grafts when re-epithelialization is slow, and the TNF-alpha blocking agent infliximab for refractory disease. Despite advances in therapy, the long-term outcome for patients with PG remains unpredictable, because relapses are common. PMID- 16372800 TI - Baseline serum levels of mast cell tryptase are raised in hemodialysis patients and associated with severity of pruritus. AB - BACKGROUND: The pathogenesis of pruritus in renal disease is not yet understood. Evidence suggests that mast cells play a role; for example, the number of dermal mast cells is increased in patients on hemodialysis. PATIENTS AND METHODS: To investigate a possible role of mast cell tryptase in pruritus of patients undergoing chronic hemodialysis, serum mast cell tryptase concentrations were measured in blood samples taken from 93 such patients, 53 of whom also recorded the severity of their pruritus on a visual analogue scale. RESULTS: Serum mast cell tryptase levels were above 11.4 microg/l (95th percentile) in 84 of 93 hemodialysis patients (90.3 %). The intensity of pruritus correlated significantly (p = 0.014) with the tryptase levels, an associated not yet shown for other mast cell-related parameters. CONCLUSIONS: Mast cells or even tryptase itself may be involved in the pathogenesis of pruritus of hemodialysis patients. PMID- 16372801 TI - [Comparison of activity of different topical corticosteroid creams and ointments using a vasoconstriction assay: superiority of hydrocortisone butyrate over hydrocortisone]. AB - BACKGROUND: Topical corticosteroids are the treatment of choice for numerous inflammatory or hyperproliferative skin diseases. The vasoconstriction assay is suitable to determinate corticosteroid activity. PATIENTS AND METHODS: 60 healthy volunteers were studied. Three corticosteroids in both cream and ointment bases, as well as one corresponding vehicle, were investigated in a double-blind, randomised, intra-individual clinical study, using a vasoconstriction assay. The potencies of hydrocortisone, hydrocortisone-17-butyrate and mometasone furoate were measured, while hydrocortisone-17-butyrate was also compared to the vehicle. RESULTS: The sum score of the clinical evaluation was for Hydrocortisone 34 (cream) and 18 (ointment), for Hydrocortisone butyrate 47 (cream) and 55 (ointment) and for Mometasone furoate 57 (cream) and 50 (ointment). The chromametric values for blanching with hydrocortisone were 1.73 (cream) and 1.48 (ointment), hydrocortisone butyrate 2.87 (cream) and 3.26 (ointment) and mometasone furoate 2.98 (cream) and 2.84 (ointment). CONCLUSIONS: The clinical and chromametric evaluation of vasoconstriction showed activity for all corticosteroid formulations. Hydrocortisone butyrate was shown to be superior to hydrocortisone. PMID- 16372802 TI - [Erythrokeratodermia variabilis (EKV)--a disorder due to altered epidermal expression of gap junction proteins]. AB - Erythrokeratodermia variabilis (EKV) is a rare autosomal dominant genodermatosis with disturbed epidermal differentiation. Its clinical picture varies from transient, fast moving erythema to persistent brown hyperkeratoses. The gene defect in EKV was recently located on the short arm of chromosome 1 encoding the gap junction protein connexin 31. We report on a 48-year-old patient with sharply circumscribed, scaling erythema on the extremities, buttocks and trunk starting since 30 years of age. Histological investigation showed orthokeratotic hyperkeratosis with focal parakeratosis overlying an acanthotic epidermis. Immunohistochemistry revealed a decreased expression of the gap junction protein connexin 31 as well as increased expression of connexin 43. At the ultrastructural level, widened intercellular spaces in the upper epidermis were present with regular desmosomes, adherens junctions and gap junctions. Epidermal cell proliferation and differentiation are regulated by gap junctions. The mutation in connexin 31 is regarded therefore as causal for the clinical picture of the EKV. The unique upregulation of connexin 43 may occur as a consequence of the Cx31 mutation and temporarily compensate for this defect. PMID- 16372803 TI - [Dental alterations in junctional epidermolysis bullosa--report of a patient with a mutation in the LAMB3-gene]. AB - During early odontogenesis the basement membrane is known to be important in epithelio-mesenchymal interactions. Mutations in the gene of one of the major structural proteins of the basement membrane such as laminin 5 might therefore be expected either to seriously compromise ameloblast differentiation and/ or interfere with normal basement-membrane formation and degradation and thus the binding of the ameloblasts to their underlying matrix. Teeth of patients suffering from junctional epidermolysis bullosa (JEB) can be severely affected by abnormal dental development and generalized or focal enamel hypoplasia. Those changes are found in 100% of individuals with JEB but the expression is variable. Beside the quantitative alterations, changes in the prismatic structure and orientation of enamel crystals are described. In addition JEB is associated with an increased risk for dental caries, caused by developmentally compromised enamel and external factors such as difficulties in maintaining oral hygiene because of oral lesions or a softer and more refined high caloric diet. Dental care includes three main strategies: Prevention by consequent oral hygiene and reduction of cariogenic nutrition is of paramount importance to minimize caries development; the restoration of enamel and dentin defects with fillings and stainless steel crowns to guarantee structure and function of teeth; and extractions of most severely affected teeth with osteolytic foci to remove continuous sources of oral infections. PMID- 16372804 TI - [Multiple fibromas in systemic mastocytosis]. AB - An adult man with systemic mastocytosis developed multiple fibromas within his involved skin, predominantly in the intertriginous areas. Friction in the intertriginous areas and scratching due to severe itch may have induced the release of mast cell factors which subsequently resulted in fibroma formation. PMID- 16372805 TI - [Systemic acne therapy]. PMID- 16372806 TI - [Painful ulcerations of both lower limbs]. PMID- 16372807 TI - [Therapy of atopic eczema with calcineurin inhibitors]. PMID- 16372808 TI - [New guidelines for rehabilitation--chances and risks for dermatology]. PMID- 16372809 TI - [Reader's letter concerning K. Kreutzer, B.Bonnekoh, I. Franke, H. Gollnick. Photodynamic therapy with methylaminooxopentanoate (Metvix) and a broad band light source (Photdyn 501): practical experiences in problem patients with actinic keratoses and basal cell carcinomas. JDDG 2004,12:992-999]. PMID- 16372810 TI - [Local anesthesia with adrenaline addition on the ear and nose]. PMID- 16372811 TI - [G-dRG version 2005: relevant aspects for dermatology]. AB - One year after the obligatory implementation of a new hospital funding system based on Diagnosis Related Groups (DRG) the third version of a German DRG-system has been published. It differs significantly from the previous version. Modifications in the classification system and the introduction of further procedure-based payments contribute to a better depiction of specialized clinical services. A number of acknowledged problems which caused great discussion in 2004 have been solved, yielding a more appropriate mapping of clinical services. The algorithms of the major diagnostic categories have been modified and complications, comorbidities and/or multiple procedures will be considered more precisely in selecting a group. However, some inconsistencies concerning highly specialized medical procedures and special features of impatient dermatological care remain. Nevertheless, a great improvement over the previous versions is apparent. The crucial aspects of the G-DRG version 2005 and the accompanying rules and regulations of payment are reviewed in detail with special relevance to dermatology. PMID- 16372812 TI - [The clinical spectrum of dermatomyositis]. AB - Dermatomyositis is characterized by marked clinical heterogeneity. Many classification schemes are available, based on internal organ involvement, presence of circulating autoantibodies and additional symptoms characteristic for other connective tissue diseases. Viral infections and underlying malignancies are possible mechanisms to trigger this autoimmune disease. Intensive clinical investigations to detect such malignancies are therefore required. The prognosis of the individual patient depends on the involvement of internal organs and the characterization of circulating autoantibodies. There are patients with rapidly developing subsets of dermatomyositis and those with slowly progressive disease. Therefore treatment has to be adjusted for the disease severity. Sometimes high dosages of corticosteroids in combination with immunosuppressive agents or immunoglobulins are required. PMID- 16372813 TI - Epinephrine-supplemented local anesthetics for ear and nose surgery: clinical use without complications in more than 10,000 surgical procedures. AB - INTRODUCTION: Local anesthetics supplemented with epinephrine are generally regarded as contraindicated for surgical procedures involving the fingers, toes, penis, outer ear and the tip of the nose [1], but epinephrine is essential if automated tumescence local anesthesia (Auto-TLA) is used. MATERIALS AND METHODS: Infiltration anesthesia supplemented with 1:200,000 epinephrine was used from 1985-1997 in our department, while Auto-TLA supplemented with 1:1.000,000 epinephrine was introduced in 1997 for all surgical procedures involving the ear or nose. During this period, 10,201 patients underwent surgery at these locations. In addition, dermal blood flow was analyzed by acral photoplethysmography (APPG) and laser Doppler flowmetry (LDF) in the right ear lobe of five normal volunteers and during epinephrine supplemented Auto-TLA. RESULTS: Epinephrine-induced complications were not observed in a single patient. Cosmetic skin flap surgery was performed in 4,953 of these patients. Even in patients with extended surgical procedures that took up to one to two hours and that included extensive skin flaps or skin grafts, we observed no increase in complications when compared to procedures performed either under general anesthesia or local anesthesia without epinephrine supplementation. Measuring blood perfusion of the earlobe showed a 69% reduction of LDF and a 42% reduction of arterial inflow (APPG) immediately following anesthesia. CONCLUSION: Epinephrine supplementation of local anesthetics does not block blood perfusion in the ear and did not induce organ, tissue or flap necrosis. Local anesthesia with epinephrine supplementation is therefore safe for acral areas such as the ear or nose. Despite the relatively small influence on blood perfusion, epinephrine supplementation results in a relatively bloodless operating field and longer effectiveness of local anesthesia. The relative absence of blood in the operating field of the ear and nose significantly reduces the duration of surgery and increases the healing rate, as less electrocautery is needed. PMID- 16372814 TI - Intravenous dexamethasone-cyclophosphamide pulse therapy in comparison with oral methylprednisolone-azathioprine therapy in patients with pemphigus: results of a multicenter prospectively randomized study. AB - BACKGROUND: Pemphigus is a potentially life-threatening autoimmune blistering skin disease usually treated with high-dose corticosteroids in combination with immunosuppressive drugs. In a multicenter, prospectively randomized study we compared efficacy and side effects of a dexamethasone-cyclophosphamide (D/C) pulse therapy with a methylprednisolone-azathioprine (M/A) therapy in 22 patients with newly diagnosed pemphigus vulgaris and pemphigus foliaceus. PATIENTS AND METHODS: The 11 patients of the M/A group were treated with daily doses of methylprednisolone (initially 2 mg/kg body weight) and azathioprine (2-2,5 mg/kg body weight) which were subsequently tapered. D/C pulse therapy in 11 patients consisted of intravenous administration of 100 mg dexamethasone/d on 3 consecutive days along with cyclophosphamide (500 mg) on day one. Pulses were initially repeated every 2-4 weeks and then at increasing intervals. In between the pulses, oral cyclophosphamide (50 mg) was given daily for 6 months. RESULTS: Within 24 months after treatment initiation, 5/11 patients of the D/C group had a remission (complete remissions after discontinuation of therapy in 3 patients) and 6/11 patients had a progression. In the M/A group, there were remissions in 9/11 patients (complete remissions after discontinuation of therapy in 3 patients) and progression in 1/11 patients. There were more relapses in M/A therapy after remission than in D/C therapy. Side effects were more common in the M/A group. These differences were not significant (p > 0,05). CONCLUSION: Because of the high number of progressions in patients treated with D/C therapy, we can not confirm the encouraging results of earlier reports about pulse D/C therapy. Nevertheless D/C therapy seemed to be better tolerated and, in case of primary efficacy, was associated with fewer recurrences than M/A therapy. PMID- 16372815 TI - [Topical treatment of bullous pemphigoid with tacrolimus. Case report with brief literature review]. AB - Bullous pemphigoid (BP) is a cutaneous autoimmune disease predominantly affecting older patients which can cause death either due to severe clinical manifestations or due to the side effects of systemic immunosuppressive treatment. Topical treatment with corticosteroids is an established alternative to systemic treatment. However, prolonged application is accompanied by side effects such as skin atrophy. Recently, the immunomodulatory calcineurin antagonists tacrolimus and pimecrolimus have been introduced for topical treatment of skin diseases. Tacrolimus has been reported to be effective in several inflammatory skin disorders such as atopic dermatitis, psoriasis, lichen planus, lupus erythematosus and pyoderma gangraenosum. Efficacy has also been described in the topical treatment of BP in some cases. Here we present the case of a 89 year old patient with BP. He was treated with 0.1% tacrolimus ointment, which was able to control the disease. We briefly review the literature and discuss the potential role of tacrolimus as a novel option for the topical treatment of BP. PMID- 16372816 TI - [Acute localized exanthematous pustulosis (ALEP)]. AB - Acute generalized exanthematic pustulosis (AGEP) is characterized by a generalized rash and sterile disseminated, sometimes coalescing subcorneal pustules. It occurs in body flexures such as the inguinal folds and intertriginous areas. The acute onset of disease is accompanied by malaise, fever > 38 degrees C and peripheral granulocytosis. We report on a female patient who according to the criteria of AGEP was diagnosed as having acute localized exanthematic pustulosis (ALEP) on the face. PMID- 16372817 TI - [Pharmacogenomics in dermatology]. PMID- 16372818 TI - [A 74-year-old lady with periorbital swelling]. PMID- 16372819 TI - [Patient information: pilonidal sinus (hair nidus dimple- or "coccyx fistula"]. PMID- 16372821 TI - Effect of genetic polymorphisms in cytochrome p450 (CYP) 2C9 and CYP2C8 on the pharmacokinetics of oral antidiabetic drugs: clinical relevance. AB - Type 2 diabetes mellitus affects up to 8% of the adult population in Western countries. Treatment of this disease with oral antidiabetic drugs is characterised by considerable interindividual variability in pharmacokinetics, clinical efficacy and adverse effects. Genetic factors are known to contribute to individual differences in bioavailability, drug transport, metabolism and drug action. Only scarce data exist on the clinical implications of this genetic variability on adverse drug effects or clinical outcomes in patients taking oral antidiabetics. The polymorphic enzyme cytochrome P450 (CYP) 2C9 is the main enzyme catalysing the biotransformation of sulphonylureas. Total oral clearance of all studied sulphonylureas (tolbutamide, glibenclamide [glyburide], glimepiride, glipizide) was only about 20% in persons with the CYP2C9*3/*3 genotype compared with carriers of the wild-type genotype CYP2C9*1/*1, and clearance in the heterozygous carriers was between 50% and 80% of that of the wild-type genotypes. For reasons not completely known, the resulting differences in drug effects were much less pronounced. Nevertheless, CYP2C9 genotype-based dose adjustments may reduce the incidence of adverse effects. The magnitude of how doses might be adjusted can be derived from pharmacokinetic studies. The meglitinide-class drug nateglinide is metabolised by CYP2C9. According to the pharmacokinetic data, moderate dose adjustments based on CYP2C9 genotypes may help in reducing interindividual variability in the antihyperglycaemic effects of nateglinide. Repaglinide is metabolised by CYP2C8 and, according to clinical studies, CYP2C8*3 carriers had higher clearance than carriers of the wild-type genotypes; however, this was not consistent with in vitro data and therefore further studies are needed. CYP2C8*3 is closely linked with CYP2C9*2. CYP2C8 and CYP3A4 are the main enzymes catalysing biotransformation of the thiazolidinediones troglitazone and pioglitazone, whereas rosiglitazone is metabolised by CYP2C9 and CYP2C8. The biguanide metformin is not significantly metabolised but polymorphisms in the organic cation transporter (OCT) 1 and OCT2 may determine its pharmacokinetic variability. In conclusion, pharmacogenetic variability plays an important role in the pharmacokinetics of oral antidiabetic drugs; however, to date, the impact of this variability on clinical outcomes in patients is mostly unknown and prospective studies on the medical benefit of CYP genotyping are required. PMID- 16372823 TI - Clinical pharmacology of lumiracoxib: a selective cyclo-oxygenase-2 inhibitor. AB - Lumiracoxib (Prexige) is a selective cyclo-oxygenase (COX)-2 inhibitor developed for the treatment of osteoarthritis, rheumatoid arthritis and acute pain. Lumiracoxib possesses a carboxylic acid group that makes it weakly acidic (acid dissociation constant [pKa] 4.7), distinguishing it from other selective COX-2 inhibitors. Lumiracoxib has good oral bioavailability (74%). It is rapidly absorbed, reaching maximum plasma concentrations 2 hours after dosing, and is highly plasma protein bound. Lumiracoxib has a short elimination half-life from plasma (mean 4 hours) and demonstrates dose-proportional plasma pharmacokinetics with no accumulation during multiple dosing. In patients with rheumatoid arthritis, peak lumiracoxib synovial fluid concentrations occur 3-4 hours later than in plasma and exceed plasma concentrations from 5 hours after dosing to the end of the 24-hour dosing interval. These data suggest that lumiracoxib may be associated with reduced systemic exposure, while still reaching sites where COX-2 inhibition is required for pain relief. Lumiracoxib is metabolised extensively prior to excretion, with only a small amount excreted unchanged in urine or faeces. Lumiracoxib and its metabolites are excreted via renal and faecal routes in approximately equal amounts. The major metabolic pathways identified involve oxidation of the 5-methyl group of lumiracoxib and/or hydroxylation of its dihaloaromatic ring. Major metabolites of lumiracoxib in plasma are the 5 carboxy, 4'-hydroxy and 4'-hydroxy-5-carboxy derivatives, of which only the 4' hydroxy derivative is active and COX-2 selective. In vitro, the major oxidative pathways are catalysed primarily by cytochrome P450 (CYP) 2C9 with very minor contribution from CYP1A2 and CYP2C19. However, in patients genotyped as poor CYP2C9 metabolisers, exposure to lumiracoxib (area under the plasma concentration time curve) is not significantly increased compared with control subjects, indicating no requirement for adjustment of lumiracoxib dose in these subjects. Lumiracoxib is selective for COX-2 compared with COX-1 in the human whole blood assay with a ratio of 515 : 1 in healthy subjects and in patients with osteoarthritis or rheumatoid arthritis. COX-2 selectivity was confirmed by a lack of inhibition of arachidonic acid and collagen-induced platelet aggregation. COX 2 selectivity of lumiracoxib is associated with a reduced incidence of gastroduodenal erosions compared with naproxen and a lack of effect on both small and large bowel permeability. Lumiracoxib does not exhibit any clinically meaningful interactions with a range of commonly used medications including aspirin (acetylsalicylic acid), fluconazole, an ethinylestradiol- and levonorgestrel-containing oral contraceptive, omeprazole, the antacid Maalox, methotrexate and warfarin (although, as in common practice, routine monitoring of coagulation is recommended when lumiracoxib is co-administered with warfarin). As such, dose adjustments are not required when co-administering these agents with lumiracoxib. In addition, moderate hepatic impairment and mild to moderate renal impairment do not appear to influence lumiracoxib exposure. PMID- 16372822 TI - Comparative pharmacokinetics of vitamin K antagonists: warfarin, phenprocoumon and acenocoumarol. AB - Vitamin K antagonists belong to the group of most frequently used drugs worldwide. They are used for long-term anticoagulation therapy, and exhibit their anticoagulant effect by inhibition of vitamin K epoxide reductase. Each drug exists in two different enantiomeric forms and is administered orally as a racemate. The use of vitamin K antagonists is complicated by a narrow therapeutic index and an unpredictable dose-response relationship, giving rise to frequent bleeding complications or insufficient anticoagulation. These large dose response variations are markedly influenced by pharmacokinetic aspects that are determined by genetic, environmental and possibly other yet unknown factors. Previous knowledge in this regard principally referred to warfarin. Cytochrome P450 (CYP) 2C9 has clearly been established as the predominant catalyst responsible for the metabolism of its more potent S-enantiomer. More recently, CYP2C9 has also been reported to catalyse the hydroxylation of phenprocoumon and acenocoumarol. However, the relative importance of CYP2C9 for the clearance of each anticoagulant substantially differs. Overall, the CYP2C9 isoenzyme appears to be most important for the clearance of warfarin, followed by acenocoumarol and, lastly, phenprocoumon. The less important role of CYP2C9 for the clearance of phenprocoumon is due to the involvement of CYP3A4 as an additional catalyst of phenprocoumon hydroxylation and significant excretion of unchanged drug in bile and urine, while the elimination of warfarin and acenocoumarol is almost completely by metabolism. Consequently, the effects of CYP2C9 polymorphisms on the pharmacokinetics and anticoagulant response are also least pronounced in the case of phenprocoumon; this drug seems preferable for therapeutic anticoagulation in poor metabolisers of CYP2C9. In addition to these vitamin K antagonists, oral thrombin inhibitors are currently under clinical development for the prevention and treatment of thromboembolism. Of these, ximelagatran has recently gained marketing authorisation in Europe. These novel drugs all feature some major advantages over traditional anticoagulants, including a wide therapeutic interval, the lack of anticoagulant effect monitoring and a low drug-drug interaction potential. However, they are also characterised by some pitfalls. Amendments of traditional anticoagulant therapy, including self-monitoring of international normalised ratio values or prospective genotyping for individual dose-tailoring may contribute to the continuous use of warfarin, phenprocoumon and acenocoumarol in the future. PMID- 16372825 TI - Pharmacokinetics and dose proportionality of fentanyl effervescent buccal tablets in healthy volunteers. AB - BACKGROUND AND OBJECTIVES: Fentanyl effervescent buccal tablets (FEBT) are designed to enhance the rate and efficiency of fentanyl absorption through the buccal mucosa. This study was undertaken to characterise the pharmacokinetics and assess the dose proportionality of FEBT in healthy volunteers within the potential therapeutic dose range. METHODS: Twenty-five healthy adults (mean age 33 years) completed a single-dose, randomised, open-label, four-dose, four period, crossover study of FEBT. They were administered FEBT 200, 500, 810 or 1080microg. The subjects in this study were not opioid tolerant; therefore, naltrexone was administered to block any opioid receptor-mediated effects of fentanyl. Venous blood samples for measurement of serum fentanyl concentrations were obtained over 36 hours following dosing. Adverse events were recorded throughout the study. RESULTS: The pharmacokinetics of FEBT were characterised by an absorption phase with a median time to reach maximum serum concentration (tmax) of 0.75-0.99 hours that was consistent irrespective of dose. Mean serum fentanyl concentrations exhibited a biexponential decline from peak after FEBT 200 and 500microg doses and a triexponential decline after FEBT 810 and 1080microg doses. The maximum serum concentration (Cmax) of fentanyl was proportional up to and including the 810microg dose. The increase in Cmax was 20% less than proportional at the 1080microg dose. Systemic exposure to fentanyl, as measured by the area under the serum concentration-time curve from time zero to infinity (AUCinfinity), increased proportionally with increasing doses of FEBT (200-1080microg). No serious adverse events were reported during the study. CONCLUSION: The pharmacokinetics of FEBT were characterised by a high early fentanyl concentration as a result of absorption across the buccal mucosa of the oral cavity, which results in bypassing first-pass metabolism. This high early tmax contributed to enhanced early systemic fentanyl exposure. Maximum concentration and AUCinfinity of FEBT increased in a dose-proportional manner from 200 to 810microg. This study provides preliminary pharmacokinetic data for FEBT across the potential therapeutic dose range. PMID- 16372824 TI - High variability of indinavir and nelfinavir pharmacokinetics in HIV-infected patients with a sustained virological response on highly active antiretroviral therapy. AB - OBJECTIVES: To describe plasma concentrations of indinavir alone or combined with ritonavir, and of nelfinavir and its active metabolite M8, and to measure their variabilities in HIV-infected patients treated with a stable antiretroviral regimen and experiencing a sustained virological response for at least 12 months. PATIENTS AND METHODS: In this prospective trial, blood samples were drawn during a 6-hour time interval between two doses at enrolment to assess protease inhibitor (PI) pharmacokinetic parameters, and 4 months later to assess plasma trough and peak concentrations. Safety and adherence assessments and laboratory data were collected during an 8-month period. PI pharmacokinetic characteristics were analysed using a non-compartmental approach. Inter- and intrapatient variabilities were estimated using a linear mixed-effect model. The impact of different covariates on plasma trough concentrations was investigated. Eighty eight patients were analysed: 42 treated with indinavir and 46 with nelfinavir. RESULTS: The interquartile range (IQR) of the plasma trough concentration corrected for the sampling time (Ccalc) was 116-374 microg/L for indinavir alone and 163-508 microg/L for indinavir/ritonavir. Ritonavir significantly increased indinavir elimination half-life and plasma exposure. For nelfinavir, the IQR of Ccalc was 896-2059 microg/L for three-times-daily administration and 998-2124 microg/L for twice-daily administration. Variabilities were high for both PIs. Intrapatient variability for indinavir alone (and indinavir + ritonavir) was 76% (107%) and interpatient variability was 58% (10%) in adherent patients. Intrapatient variability for nelfinavir three times daily (and twice daily) was 41% (74%) and interpatient variability was 62% (50%). Intrapatient variability was lowered in patients with a high adherence level. CONCLUSION: Although performed in a homogeneous population, this study documented a high interpatient but also intrapatient variability of indinavir and nelfinavir pharmacokinetics, which should be taken into account when interpreting therapeutic drug monitoring. Once patients have reached a sustained virological response, plasma PI monitoring may have a limited impact. PMID- 16372826 TI - Penetration of orally administered prulifloxacin into human lung tissue. AB - OBJECTIVE: To evaluate the distribution in lung tissue of ulifloxacin, the active metabolite of prulifloxacin, a new once-daily fluoroquinolone administered orally in a single 600mg dose. DESIGN: Open-label, randomised study. PATIENTS: Twenty seven patients (25 males, 2 females; mean age 65.7 years [range 49-79 years]) with a lung neoplasm requiring lobectomy or pneumonectomy. METHODS: Patients were randomly assigned to five treatment groups and received a single oral dose of prulifloxacin 600mg at 2, 4, 6, 12 or 24 hours preoperatively. During surgery, blood and healthy lung (based on macroscopic appearance) samples were collected at the same time. Ulifloxacin concentrations in plasma and lung tissue were determined by a validated reversed-phase high-performance liquid chromatography assay. Lung tissue ulifloxacin concentrations were adjusted for blood contamination, by measuring haemoglobin in the supernatant of each tissue sample and applying a corrective equation. RESULTS: Ulifloxacin concentration in lung tissue exceeded plasma concentration at every timepoint. Following administration of prulifloxacin 600mg, the overall mean corrected lung/plasma ratio over the 24 hour period was 6.9 (range 1.2-14.1). When sampling intervals were assessed, the corrected lung/plasma ratios were 7.5 (2 hours after dosing), 6.3 (4 hours), 4.3 (6 hours), 7.0 (12 hours) and 9.2 (24 hours). The mean corrected lung/plasma area under the concentration-time curve ratio was 6.3, demonstrating the ability of the drug to penetrate lung tissue and confirming the high exposure of this target tissue to ulifloxacin. However, the limitation of the lung tissue sampling method and the high interpatient variability should be considered. Over the 24-hour period, the concentrations of ulifloxacin in lung tissue were higher than the minimum inhibitory concentration (MIC) values for pathogens frequently involved in community-acquired respiratory tract infections. CONCLUSION: Lung tissue penetration data may have a supportive value when considered jointly with MICs and efficacy results. The findings from this lung penetration study could explain the efficacy of once-daily prulifloxacin 600mg observed in clinical trials conducted in patients with exacerbation of chronic bronchitis. PMID- 16372827 TI - Population pharmacokinetics of onercept in healthy subjects. AB - OBJECTIVE: To develop a population pharmacokinetic model and to determine the covariates affecting the pharmacokinetics of onercept (recombinant human tumour necrosis factor [TNF] receptor-1) in healthy subjects. SUBJECTS AND METHODS: Onercept pharmacokinetics data were obtained from 48 healthy male and female subjects (four phase I studies). In study A, 12 subjects received increasing single intravenous doses of onercept either 5 and 50mg or 15 and 150mg. In study B, 12 subjects received single intravenous, subcutaneous and intramuscular doses of onercept 50mg. Study C investigated the pharmacokinetics of onercept following repeat subcutaneous administration of six doses of 50mg every 48 hours in 12 subjects. Study D investigated the pharmacokinetics of onercept following repeat subcutaneous administration of six doses of 100mg and 150mg over 2 weeks in 12 subjects. Nonlinear mixed-effects modelling software NONMEM was used to build a base model, while the final model was determined after selection of the covariates. RESULTS: The disposition of onercept was described using a two compartment model with two absorption processes (a first-order followed by a zero order) and included a constant baseline, accounting for the endogenous TNF receptor-1 levels. Slow absorption of onercept following subcutaneous and intramuscular administration was observed and suggested that absorption was the rate-limiting process. The population mean (coefficient of variation %) values for clearance, absorption rate constant, volume of distribution of the central compartment, bioavailability of onercept and baseline TNF receptor-1 levels were 4.03 L/h (13.3%), 0.04 h-1 (29.1%), 4.42L (6.2%), 0.90 (23.8%) and 1.68 microg/L (20.4%), respectively. The only significant covariates were found to be dose (which affected clearance), and day (which affected absorption rate constant); however, the effects were small (10-15%) and are unlikely to be of any clinical relevance. CONCLUSION: The proposed population pharmacokinetic model characterises well the overall pharmacokinetic profile of onercept after intramuscular, subcutaneous and intravenous administration in healthy subjects. The pharmacokinetics of onercept showed modest intersubject variability. PMID- 16372828 TI - Cystatin C as a new covariate to predict renal elimination of drugs: application to carboplatin. AB - BACKGROUND AND OBJECTIVE: The individual dosing of drugs that are mainly eliminated unchanged in the urine is made possible by assessing renal function. Most of the methods used are based on serum creatinine (SCr) levels. Cystatin C (CysC) has been proposed as an alternative endogenous marker of the glomerular filtration rate (GFR). Carboplatin is one of the drugs for which elimination is most dependent on the GFR. A prospective clinical trial including 45 patients was conducted to assess the value of serum CysC as a predictor of carboplatin clearance (CL). METHODS: The patients were receiving carboplatin as part of established protocols. Carboplatin was administered as a daily 60-minute infusion at doses ranging from 290 to 1700mg. A population pharmacokinetic analysis was performed using the nonlinear mixed effect modelling NONMEM program according to a two-compartment pharmacokinetic model. RESULTS: Data from 30 patients were used to test the relationships between carboplatin CL and morphological, biological and demographic covariates previously proposed for prediction of the GFR. The interindividual variability of carboplatin CL decreased from 31% (no covariate) to 14% by taking into account five covariates (SCr, CysC, bodyweight [BW], age and sex). Prospective evaluation of these relationships using the data from the other 15 patients confirmed that the best equation to predict carboplatin CL was based on these five covariates, with a mean absolute percentage error of 13% as an assessment of precision. NONMEM analysis of the whole dataset (n = 45 patients) was performed. The best covariate equation corresponding to the overall analysis was: CL (mL/min) = 110 x (SCr/75)-0.512 x (CysC/1.0)-0.327 x (BW/65)0.474 x (age/56)-0.387 x 0.854sex, with SCr in micromol/L, CysC in mg/L, BW in kilograms, age in years and sex = 0 if male and 1 if female. To put the value of CysC as an endogenous marker of the GFR into perspective, covariate equations without SCr were also evaluated; a better prediction was obtained by considering CysC together with age and BW (interindividual variability of 16.6% vs 23.3% for CysC alone). CONCLUSION: CysC is a marker of drug elimination that is at least as good as SCr for predicting carboplatin CL. The model based on five covariates was superior to those based on only four covariates (with BW, age and sex combined with either SCr or CysC), indicating that CysC and SCr are not completely redundant to each other. Further pharmacokinetic evaluation is needed to determine whether SCr or CysC is the better marker of renal elimination of other drugs. PMID- 16372829 TI - Pharmacokinetic study of tacrolimus in cystic fibrosis and non-cystic fibrosis lung transplant patients and design of Bayesian estimators using limited sampling strategies. AB - OBJECTIVES: To: (i) test different pharmacokinetic models to fit full tacrolimus concentration-time profiles; (ii) estimate the tacrolimus pharmacokinetic characteristics in stable lung transplant patients with or without cystic fibrosis (CF); (iii) compare the pharmacokinetic parameters between these two patient groups; and (iv) design maximum a posteriori Bayesian estimators (MAP-BE) for pharmacokinetic forecasting in these patients using a limited sampling strategy. METHODS: Tacrolimus blood concentration-time profiles obtained on three occasions within a 5-day period in 22 adult lung transplant recipients (11 with CF and 11 without CF) were retrospectively studied. Three different one compartment models with first-order elimination were tested to fit the data: one with first-order absorption, one convoluted with a gamma distribution to describe the absorption phase, and one convoluted with a double gamma distribution able to describe secondary concentration peaks. Finally, Bayesian estimation using the best model and a limited sampling strategy was tested in the two groups of patients for its ability to provide accurate estimates of the main tacrolimus pharmacokinetic parameters and exposure indices. RESULTS: The one-compartment model with first-order elimination convoluted with a double gamma distribution gave the best results in both CF and non-CF lung transplant recipients. The patients with CF required higher doses of tacrolimus than those without CF to achieve similar drug exposure, and population modelling had to be performed in CF and non-CF patients separately. Accurate Bayesian estimates of area under the blood concentration-time curve from 0 to 12 hours (AUC12), AUC from 0 to 4 hours, peak blood concentration (Cmax) and time to reach Cmax were obtained using three blood samples collected at 0, 1 and 3 hours in non-CF patients (correlation coefficient between observed and estimated AUC12, R2 = 0.96), and at 0, 1.5 and 4 hours in CF patients (R2 = 0.91). CONCLUSION: A particular pharmacokinetic model was designed to fit the complex and highly variable tacrolimus blood concentration-time profiles. Moreover, MAP-BE allowing tacrolimus therapeutic drug monitoring based on AUC12 were developed. PMID- 16372830 TI - Population pharmacokinetics of olmesartan following oral administration of its prodrug, olmesartan medoxomil: in healthy volunteers and hypertensive patients. AB - BACKGROUND: Olmesartan medoxomil (CS-866) is a new orally active angiotensin II receptor antagonist that is highly selective for the AT1 receptor subtype. OBJECTIVE: To develop a population pharmacokinetic model for olmesartan (RNH 6270), the active metabolite of olmesartan medoxomil, in healthy volunteers and hypertensive patients, and to evaluate effects of covariates on the apparent oral clearance (CL/F), with particular emphasis on the effect of race. DESIGN: Retrospective analysis of data from 12 phase I-III trials in the US, Europe and Japan. PARTICIPANTS: Eighty-nine healthy volunteers and 383 hypertensive patients. METHODS: Nonlinear mixed-effects modelling was used to evaluate 7911 olmesartan plasma sample concentrations. The covariates included age, bodyweight, sex, race (Westerners [including Caucasians and Hispanics] versus Japanese), patient status (hypertensive patients versus healthy volunteers), serum creatinine level as an index of renal function and serum chemistry data as indices of hepatic function. RESULTS: The pharmacokinetic data of olmesartan were well described by a two-compartment linear model with first-order absorption and an absorption lag-time, parameterised in terms of CL/F (6.66 L/h for a typical male Western hypertensive patient), absorption rate constant (1.46h-1), elimination rate constant (0.193h-1), rate constant from the central to peripheral compartment (0.061h-1), rate constant from the peripheral to central compartment (0.079h-1) and absorption lag-time (0.427h). Analysis of covariates showed that age, bodyweight, sex, patient status and renal function were factors influencing the clearance of olmesartan. CONCLUSION: The population pharmacokinetic analysis of olmesartan showed that: (i) severe renal impairment (serum creatinine >265 micromol/L [approximately 3 mg/dL]) could cause a clearance decrease of > or =30%; (ii) older age, lower bodyweight and being female were determinants of lower clearance but their effects on olmesartan clearance were within 20%; (iii) no statistically significant difference in clearance was found between Westerners and Japanese. PMID- 16372831 TI - "Europe goes alternative"--what's all the fuss about? PMID- 16372832 TI - Heterologous co-expression of human cytochrome P450 1A2 and polymorphic forms of N-acetyltransferase 2 for studies on aromatic amines in V79 Chinese hamster cells. AB - V79 Chinese hamster cells were genetically engineered for the stable co expression of human cytochrome P450 1A2 and the polymorphic N-acetyltransferase 2 alleles *4, *5B, *6A and *13, in order to generate an in vitro tool for studying the metabolism-dependent toxicity of aromatic amines. N-acetyltransferase 2*4 encoding cDNA was generated by the polymerase chain reaction (PCR) with defined primers from the genomic DNA of a human liver donor homozygous for *4, and served as a template to generate the *5B, *6A and *13 isoforms by site-directed mutagenesis. Human cytochrome P450 (CYP) 1A2-encoding cDNA was generated by the PCR from genomic DNA of the recombinant V79MZh1A2 cell line. All the cDNAs were inserted into a CMV promoter-containing plasmid in conjunction with the selectable marker genes, neomycin and hydromycin. The recombinant expression plasmids were transfected for stable integration into the genomic DNA of the V79 cells. Several cellular clones were obtained and checked for the genomic integration of intact cDNAs with the PCR on the genomic DNA of the recombinant cells. Stable expression was confirmed by the reverse transcriptase PCR (RT-PCR) on RNA preparations. Metabolic function was tested with ethoxyresorufin as a marker substrate for CYP1A2, and 2-aminofluorene and N-sulphametazine for N acetyltransferase activity, and compared to data obtained from biological samples. 7-Ethoxyresorufin-O-deethylase activities ranged from 0.2 to 4 pmol resorufin/min/mg total protein. The N-acetylation of sulphametazine ranged from 0.07 to 1.7 nmol N-acetyl-sulphametazine/mg total protein/min. Selected clones showing activities in the range of physiological activities were submitted to metabolism dependent mutagenicity studies. In particular, the polymorphism dependent N-acetylation of 2-aminofluorene and the role of CYP1A2 and N acetyltransferase in the mutagenicity of 2-aminofluorene, were investigated. Surprisingly, the mutagenicity of 2-aminofluorene is dramatically reduced in V79 cells co-expressing CYP1A2 and N-acetyltransferase, compared to V79 cells expressing CYP1A2 only, pointing to a significant species-dependent difference in the metabolic activation of aromatic amines between rats and humans. PMID- 16372833 TI - Detection of electrophysiological indicators of neurotoxicity in human and rat brain slices by a three-dimensional microelectrode array. AB - Electrophysiological techniques for the assessment of in vitro neurotoxicology have several advantages over other currently-used methods (for example, morphological techniques), including the ability to detect damage at a very early stage. Novel recording techniques based on microelectrode arrays are available, and could improve recording power. In this study, we investigated how a three dimensional microelectrode array detects the electrophysiological endpoints of neurotoxicity. We conclude that electrophysiology sensitively reveals neurotoxic actions, and that three-dimensional microelectrode arrays could be proposed for use in neurotoxicology as recording tools that allow easy and sensitive multisite recording, from both rodent and human brain tissue. PMID- 16372834 TI - An evaluation of the in vitro cytotoxicities of 50 chemicals by using an electrical current exclusion method versus the neutral red uptake and MTT assays. AB - According to the 2001 National Institutes of Health guidance document on using in vitro data to estimate in vivo starting doses for acute toxicity, the performance of the electrical current exclusion method (ECE) was studied for its suitability as an in vitro cytotoxicity test. In a comparative study, two established in vitro assays based on the quantification of metabolic processes necessary for cell proliferation or organelle integrity (the MTT/WST-8 [WST-8] assay and the neutral red uptake [NRU] assay), and two cytoplasm membrane integrity assays (the trypan blue exclusion [TB] and ECE methods), were performed. IC50 values were evaluated for 50 chemicals ranging from low to high toxicity, 46 of which are listed in Halles Registry of Cytotoxicity (RC). A high correlation was found between the IC50 values obtained in this study and the IC50 data published in the RC. The assay sensitivity was highest for the ECE method, and decreased from the WST-8 assay to the NRU assay to the TB assay. The consistent results of the ECE method are based on technical standardisation, high counting rate, and the ability to combine cell viability and cell volume analysis for detection of the first signs of cell necrosis and damage of the cytoplasmic membrane caused by cytotoxic agents. PMID- 16372835 TI - An inter-laboratory study to evaluate the effects of medium composition on the differentiation and barrier function of Caco-2 cell lines. AB - Differentiated human intestinal Caco-2 cells are frequently used in toxicology and pharmacology as in vitro models for studies on intestinal barrier functions. Since several discrepancies exist among the different lines and clones of Caco-2 cells, comparison of the results obtained and optimisation of models for use for regulatory purposes are particularly difficult, especially with respect to culture conditions and morphological and biochemical parameters. An inter laboratory study has been performed on the parental cell line and on three clonal Caco-2 cell lines, with the aim of standardising the culture conditions and identifying the best cell line with respect to parameters relevant to barrier integrity, namely, trans-epithelial electrical resistance (TEER) and mannitol passage, and of epithelial differentiation (alkaline phosphatase activity). Comparison of the cell lines maintained in traditional serum-supplemented culture medium or in defined medium, containing insulin, transferrin, selenium and lipids, showed that parameter performance was better and more reproducible with the traditional medium. The maintenance of the cell lines for 15 days in culture was found to be sufficient for the development of barrier properties, but not for full epithelial differentiation. Caco-2/TC7 cells performed better than the other three cell lines, both in terms of reproducibility and performance, exhibiting low TEER and mannitol passage, and high alkaline phosphatase activity. PMID- 16372836 TI - An inter-laboratory collaborative study by the Non-Genotoxic Carcinogen Study Group in Japan, on a cell transformation assay for tumour promoters using Bhas 42 cells. AB - The Bhas promotion assay is a cell culture transformation assay designed as a sensitive and economical method for detecting the tumour-promoting activities of chemicals. In order to validate the transferability and applicability of this assay, an inter-laboratory collaborative study was conducted with the participation of 14 laboratories. After confirmation that these laboratories could obtain positive results with two tumour promoters, 12-O tetradecanoylphorbol-13-acetate (TPA) and lithocholic acid (LCA), 12 coded chemicals were assayed. Each chemical was tested in four laboratories. For eight chemicals, all four laboratories obtained consistent results, and for two of the other four chemicals, only one of the four laboratories showed inconsistent results. Thus, the rate of consistency was high. During the study, several issues were raised, each of which were analysed step-by-step, leading to revision of the protocol of the original assay. Among these issues were the importance of careful maintenance of mother cultures and the adoption of test concentrations for toxic chemicals. In addition, it is suggested that three different types of chemicals show positive promoting activity in the assay. Those designated as T-type induced extreme growth enhancement, and included TPA, mezerein, PDD and insulin. LCA and okadaic acid belonged to the L-type category, in which transformed foci were induced at concentrations showing growth-inhibition. In contrast, M-type chemicals, progesterone, catechol and sodium saccharin, induced foci at concentrations with little or slight growth inhibition. The fact that different types of chemicals similarly induce transformed foci in the Bhas promotion assay may provide clues for elucidating mechanisms of tumour promotion. PMID- 16372837 TI - Opportunities to replace the use of animals in sepsis research. The report and recommendations of a Focus on Alternatives workshop. AB - Sepsis and multiple organ failure are common causes of death in patients admitted to intensive care units. The incidence of sepsis and associated mortalities has been steadily increasing over the past 20 years. Sepsis is a complex inflammatory condition, the precise causes of which are still poorly understood. Animal models of sepsis have the potential to cause substantial suffering, and many of them have been poorly representative of the human syndrome. However, a number of non animal approaches, including in vitro, in silico and clinical studies, show promise for addressing this situation. This report is based on discussions held at an expert workshop convened by Focus on Alternatives and held in 2004 at the Wellcome Trust, London. It provides an overview of some non-animal approaches to sepsis research, including their strengths and weaknesses, and argues that they should be prioritised for further development. PMID- 16372839 TI - Borderline defenses and Rorschach responses: a critique of Lerner, Albert, and Walsh. AB - Collapsing DSM-III schizotypal and borderline groups to form a borderline population is unwarranted when the results are then related to borderline personality organization. In a comparison of overlapping scoring systems for inferring defensive operations, Lerner, Albert, and Walsh intertwine fact and theory extravagantly and are not justified by their chosen populations nor their assumptions about the theories involved. PMID- 16372840 TI - Toward a four-factor theory of temperament and/or personality. AB - This article makes a case for a four-factor theory of temperament/personality, tracing its origin to Hippocrates, who first postulated a four-factor theory of temperament in 400 B.C. Historical developments are discussed as they relate to the postulation of a four-factor mode, continuing from Galen in the 2nd century A.D. to the present day. Theoretical formulations are discussed along with considerations of applications in the field of personality assessment. PMID- 16372841 TI - Personality characteristics of Hare Krishnas. AB - Personality characteristics of 132 males and 94 females of the Hare Krishna movement were assessed using the Comrey Personality Scales (CPS). Subjects were sampled from eight U.S. sites. Their age averaged about 30 years, and their time in the movement averaged 8.6 years. The most prominent and surprising finding is the hallmark characteristic of the Hare Krishna personality, a strong compulsivity trait common to both genders, which is slightly stronger in males. Reduced trust in society was exhibited by both gender averages, with slightly less trust among females. Both these averages, however, were within the normal range for individuals. With the exception of compulsivity, average male CPS scores were within the normal psychological range but differed significantly from the normative male group indicating idiosyncratic traits. Average female CPS scores did not differ significantly from the normative female group except for compulsivity and trust. Study of CPS score distributions about group means supported these findings. PMID- 16372842 TI - The Spanish version of the Clinical Analysis Questionnaire: a precautionary note. AB - The Spanish version of the Clinical Analysis Questionnaire (CAQ) was examined and found to have numerous grammatical and syntactical errors that seriously limit its usefulness for the psychological evaluation of Spanish-speaking clients. At least 36% of the items were found to be poorly translated from the original English version. PMID- 16372843 TI - A redundancy analysis for the Inwald Personality Inventory and the MMPI. AB - A redundancy analysis between two psychological preemployment screening inventories was conducted with 2,438 male correction officer candidates. One canonical variate accounted for the majority of overlapping variance. When cross validated, redundancy indices were. 17 for the Inwald Personality Inventory (IPI) and. 15 for the Minnesota Multiphasic Personality Inventory (MMPI). Three areas of overlap-control over actions, external control, and restlessness-were suggested. Less pathological behavioral patterns predictive of future job performance appeared measured only by the IPI. These results, combined with results from predictive validity studies, provide evidence that pathology is not sufficient for identifying "high risk" candidates. PMID- 16372844 TI - The convergent and discriminant validity of the Need for Cognition Scale. AB - This article reports two studies that examined the convergent and discriminant validity of the Need for Cognition Scale (Cacioppo & Petty, 1982). Using samples of college students and prison inmates, need for cognition scores were found to be positively associated with measures of self-esteem, masculine sex role attitudes, absorption, and private self-consciousness. Modest negative associations between need for cognition and measures of public self-consciousness and social anxiety were also uncovered. Need for cognition scores were generally unrelated to measures of feminine and androgynous sex role attitudes, shyness, sociability, and loneliness. These findings add further evidence supporting the construct validity of the Need for Cognition Scale and expand our understanding of the construct of need for cognition. PMID- 16372846 TI - A scale for measuring persistence in children. AB - Persistence and its assessment have been studied since the 1920s. One common approach in measuring persistence has been through testing the subject's ability to solve difficult or monotonous problems. Another approach has been through questionnaires. Although persistence is very important in academic achievement and in the general development of personality, no questionnaires have been developed for measuring persistence in children. The present research concerns the development of such a scale. It includes 40 items and was tested on 322 Israeli children aged 7-13. The internal reliability of the scale (Cronbach alpha) was .66, and the test-retest reliability after 6 months was .77. The scale differentiated active gymnasts from non-gymnasts as well as persistent gymnasts from dropout gymnasts. In addition, significant correlations were found between the scale and the need-persistence measure of the Rosenzweig Picture Frustration Study, Locus of Control, and other measures. We discuss these findings along with the significance of persistence in everyday life. PMID- 16372847 TI - The Belief in Personal Control Scale: a measure of God-Mediated and Exaggerated Control. AB - Combining and extending previous conceptions of the construct of perceived control, a self-report measure based on a proposed multidimensional model was developed and validated. The Belief in Personal Control Scale (BPCS), measures of locus of control, depression, mania, anxiety, self-esteem, and social desirability were administered to several samples. A factor analysis revealed three interpretable components of the control construct: a general External Control factor, an Exaggerated Internal Control factor, and a God-Mediated Control factor. The results of this article provide preliminary support for the model and for the BPCS as a multidimensional measure of perceived control with reliable and valid subscales. Consistent with the proposed model of control, the three factors correlated differentially with the various measures of psychological adjustment. Of particular interest is the positive correlation between the Exaggerated Internal Control factor and a measure of mania. PMID- 16372848 TI - Development of a short-form Sex Role Behavior Scale. AB - This article describes the construction of a short-form of the Sex Role Behavior Scale (SRBS). The SRBS is a self-report inventory that samples the interest/behavior domain of sex roles comprehensively. It consists of male-valued (M), female-valued (F), and sex-specific (MF) interests and behaviors in four areas: leisure activity preferences; vocational interests; social interaction; and marital, or primary relationship, behaviors. We administered the SRBS to 411 college men and women along with measures of sex role personality traits and attitudes. Items were selected from the SRBS for inclusion in the short form. We examined male-female differences on the short-form scales, internal consistency, intercorrelations among the scales, and relations of the short-form scales with the long form and with measures of sex role traits and attitudes. The results indicate that the short-form SRBS is a satisfactory substitute for the lengthy inventory from which it was drawn. However, limited internal consistency for some of the separate behavior area subscales suggests that the short form is most suitable for global assessments of sex role interests/behaviors, whereas the long form is more appropriate when more specific assessments are required in the separate interest/behavior areas represented on the SRBS. PMID- 16372851 TI - Norms for college undergraduates for the Bem Sex-Role Inventory and the Wiggins Interpersonal Behavior Circle. AB - Norms based on large random samples of introductory psychology college students are presented for the Bem Sex-Role Inventory (N = 1,133) and the Wiggins Interpersonal Behavior Circle (N = 564). The use of normative data, as compared with raw data or data standardized separately within studies, should result in more precise and meaningful measurement of subjects' locations on sex-role dimensions (i.e., instrumentality and expressiveness) by future researchers using these instruments, although norms should be updated to reflect future research results. Conventional definitions of moderately extreme and statistically extreme scores are presented for these instruments based on these norms, encouraging standardization of sample characteristics of future studies. PMID- 16372852 TI - Ego identity and desperate love. AB - The relationships between ego-identity consolidation, a romantic attitude toward love, and a desperate style of fusional love relations were examined using several questionnaire measures: Tan, Kendis, Fine, and Porac's (1977) Ego Identity Scale, Sperling's (1985) Desperate Love Scale, and Knox and Sporakowski's (1968) Attitude Toward Love Scale. Results based on an initial undergraduate sample of 84 men and 87 women, and a follow-up sample of 116 men, supported the primary hypothesis of a negative correlation between ego-identity consolidation and desperate love, as well as the secondary hypotheses. PMID- 16372853 TI - An Authority Behavior Inventory. AB - This article describes the development and validation of an Authority Behavior Inventory (ABI) to assess acceptance of authority. Each of the 24 items on the self-report ABI correlated significantly with corresponding peer ratings. In two studies (n = 200 and n = 150), the ABI was shown to have high internal consistency and high degrees of concurrent validity. Validity was assessed by means of correlations with the General Attitude Towards Institutional Authority Scale (Rigby, 1982, 1984a) using a student and a nonstudent sample; and, among nonstudents, with ratings of attitude to authority provided independently by both the subjects and their raters. In addition, the ABI discriminated significantly between matched subjects judged by peers as differing in attitude to authority, and between supporters of two Australian political parties differing in conservatism. PMID- 16372857 TI - Distinguishing thinking-feeling preferences through the content analysis of natural language. AB - This article investigates the relationship between natural language and personality characteristics. In particular, an effort was made to determine whether "thinkers" and "feelers," as defined by Jung's typologies, differ in terms of the language they use to express themselves. First, a linguistic content analysis scale was developed. Next, 5-min verbal samples were obtained from subjects and content analyzed. Content analysis scores were correlated with subjects' self-evaluations and their scores on the Thinking-Feeling scale of the Myers-Briggs Type Indicator (MBTI). Gender differences relating to thinking feeling preferences were also investigated. Significant correlations between content analysis and MBTI scores were obtained, suggesting that valid information regarding individual preferences for thinking versus feeling may be obtained through the analysis of natural language. Males and females differed significantly in terms of their MBTI scores and self-evaluations, but not in their content analysis scores. PMID- 16372858 TI - Testing the dimensionality of the self-consciousness scales. AB - The self-consciousness scale of Fenigstein, Scheier, and Buss (1975) was subjected to internal and external consistency tests based on the classical test theory model. The scale was found to have five underlying dimensions: two for private self-consciousness (viz., self-reflectiveness and internal state awareness), two for public self-consciousness, and one for social anxiety. The confirmatory factor analysis procedures employed by Burnkrant and Page (1984) are shown to be fallible as indicators of unidimensionality. Theoretical implications of newly identified dimensions in the public self-consciousness subscale are discussed. PMID- 16372859 TI - Computer-generated psychological reports: a look at the modern psychometric machine. AB - The proliferation of computer-generated psychological reports prompts an examination of the presuppositions of such reports and what they offer in the way of understanding individual clients. This topic is approached primarily through a review of some basics of psychological testing and psychological assessment. Major differences are noted between these two approaches to evaluating clients: They are different processes, and they yield different products. Assessment is a complex set of activities that seeks solutions to specific problems, whereas testing fits a straightforward actuarial paradigm. Testing is readily adapted to an automated data processing system, whereas the complex of assessment procedures cannot be so managed. Serious questions about the validity of computer reports are raised, however, and ethical concerns are expressed over the use of these reports by persons without adequate psychological training to understand them correctly. Limited uses for computer reports are suggested along with the need for further studies of validity and ethics. PMID- 16372860 TI - Keep 'Gestalt' in the name of the test: the curious name permutations of the Bender-Gestalt Test. AB - The Bender-Gestalt Test is often referred to in the literature by several different names, or just as the Bender Test, without the term Gestalt. As a consequence, the figures of the test are frequently called Bender figures or Bender designs, terms which are unjustified and misleading because the figures have been taken from Wertheimer's classical study of Gestalt laws. In this article, I trace the beginnings of the name of the test and its curious permutations, and I advocate the retention of the term Gestalt in the name of the test for logical and historical reasons. PMID- 16372861 TI - The interpreter's personality in Draw-A-Person interpretation: a study of interpersonal style. AB - This study investigated the relationship between accuracy in figure drawing interpretation and interpersonal style. Subjects were 66 undergraduates who completed the Minnesota Multiphasic Personality Inventory (MMPI), the Interpersonal Check List (ICL), and a figure drawing interpretation task. The MMPI and ICL were scored and interpreted using Leary's Interpersonal System, which provides for a multilevel assessment of interpersonal functioning. Interpretive skill was found to be associated with "affiliative" interpersonal styles (p < .05), and lack of interpretive skill was associated with "disaffiliative" interpersonal styles (p < .05) at two of the three levels examined. The role of unconscious processes in figure drawing interpretation, the nature of the interpretive process, and implications for the selection of skilled interpreters are discussed with reference to the interpersonal characteristics of the interpreter. PMID- 16372862 TI - Assessing the person reliability of an individual MMPI protocol. AB - This study investigated various measures commonly employed to assess the person reliability of an individual Minnesota Multiphasic Personality Inventory (MMPB protocol. Specifically, relationships among indices of person reliability and the standard MMPI validity scales were examined using the responses of 82 subjects who completed the MMPI on two occasions separated by 1 week. Person reliability indices were based on within-occasion responses to identical and to psychologically similar items, and on three across-occasion response consistency measures. The validity scales, namely, the L, F, K, and Cannot Say scales, showed higher test-retest stability than the within-occasion person reliability indices. Further, the validity scales and person reliability indices appeared to reflect multiple facets of dependable responding. Interestingly, an individual's tendency to change responses to MMPI items from the test to the retest was significantly predictable. Clinical implications of these findings were derived. PMID- 16372863 TI - SPECIAL SERIES: advances in statistical methods for personality assessment research. PMID- 16372864 TI - Statistical methods in personality assessment research. AB - Emerging models of personality structure and advances in the measurement of personality and psychopathology suggest that research in personality and personality assessment has entered a stage of advanced development, in this article we examine whether researchers in these areas have taken advantage of new and evolving statistical procedures. We conducted a review of articles published in the Journal of Personality, Assessment during the past 5 years. Of the 449 articles that included some form of data analysis, 12.7% used only descriptive statistics, most employed only univariate statistics, and fewer than 10% used multivariate methods of data analysis. We discuss the cost of using limited statistical methods, the possible reasons for the apparent reluctance to employ advanced statistical procedures, and potential solutions to this technical shortcoming. PMID- 16372865 TI - Logistic regression. AB - Logistic regression has probably been underutilized in clinical investigations of personality because of its relatively recent development (dictated by the need for computer programs to obtain maximum likelihood estimates), and the fact that use has been largely confined to the fields of biostatistics, epidemiology, and economics Its use should be given serious consideration when the outcome of interest is dichotomous (or polychotomous) in nature and the predictors of interest may be categorical or continuous. The logit transformation is quite tractable mathematically, and it embodies the notion of threshold, which may have relevance for many of the variables that are of interest to investigators in the field of personality. Furthermore, investigators with experience in multiple linear regression or contingency table analysis should have little trouble in transitioning to logistic regression. Logistic regression programs are readily available in the major statistical packages, all of which provide fairly standard output. PMID- 16372866 TI - Exploratory factor analysis: its role in item analysis. AB - The special characteristics of items-low reliability, confounds by minor, unwanted covariance, and the likelihood of a general factor-and better understanding of factor analysis means that the default procedure of many statistical packages (Little Jiffy) is no longer adequate for exploratory item factor analysis. It produces too many factors and precludes a general factor even when that means the factors extracted are nonreplicable. More appropriate procedures that reduce these problems are presented, along with how to select the sample, sample size required, and how to select items for scales. Proposed scales can be evaluated by their correlations with the factors; a new procedure for doing so eliminates the biased values produced by correlating them with either total or factor scores. The role of exploratory factor analysis relative to cluster analysis and confirmatory factor analysis is noted. PMID- 16372867 TI - Factor analytic approaches to personality item-level data. AB - Factor analysis models have played a central role in formulating conceptual models in personality and personality assessment, as well as in empirical examinations of personality measurement instruments. Yet, the use of item-level data presents special problems for factor analysis, applications. In this article, we review recent developments in factor analysis that are appropriate for the type of item-level data often collected in personality. Included in this review are discussions of how these developments have been addressed in the context of two different (but formally related) statistical models item response theory (IRT: Hambleton, Swaminathan, & Rogers, 1991) and structural, equation modeling (Bollen 1989) for item-level data. We also discuss the relevance of item scaling in the context of these models. Using the restandardization data for the Minnesota Multiphasic Personality Inventory-2 Scale (cf. Butcher, Dahlstrom, Graham, Tellegen, & Kaemmer, 1989), we show brief examples of the utility of these approaches to address basic questions about responses to personality scale items regarding: (a) scale, dimensionality and general item properties, (b) the "appropriateness" of the observed responses, and (c) differential item functioning across subsamples. implications for analyses of personality item level data in the IRT and factor analytic traditions are discussed. PMID- 16372868 TI - Problems in Rorschach research and what to do about them. AB - This article identifies many interrelated problems that interfere with progress in Rorschach research. These include vague hypotheses, confounded comparisons, failure to control extraneous variables; misuse of statistics, and distorted interpretation of results. Two crucial solutions to address many of these problems are offered: knowing the data and paling out rival hypotheses. Underlying conceptual confusions are addressed and prototypical examples derived from the literature, are discussed is the hope of improving the scientific foundation of the Rorschach. PMID- 16372869 TI - The Washington University Sentence Complete Test: are the two halves alternate forms? Are the female and male forms comparable? AB - Evidence from 85 adult medical outpatients supported psychometric comparability of the 2 halves of the Washington University Sentence Completion Test (SCT) Form 81 and of the female and male forms of the SCT. There was slightly stronger internal consistency for the first versus the second half of the SCT. Each half correlated highly with the ogive total protocol rating and 36-item-sum rating. Intercorrelations of the 2 halves with external measures also suggested essentially equivalent relations. For the 30 identical items across gender, the median correlation between individual item ratings with the item-sum ratings was nearly equal for women and men. When the 6 nonidentical items were considered with the identical items, the median item-total correlation was slightly higher for men (45) than women (41). This difference was accounted for by the slightly larger variability in the mate subsample. Practically speaking, the 2 halves and the female and male forms may be used with minimal concern regarding psychometric comparability in similar medical outpatient settings. PMID- 16372872 TI - Beneficial and adverse side effects of vaccination: perception in developing countries. PMID- 16372875 TI - A combined diphtheria, tetanus, five-component acellular pertussis, poliovirus and Haemophilus influenzae type b vaccine. AB - Ideally, combination vaccines should not only be safe and effective, but also integrate smoothly into the vaccination schedule and provide advantages over the use of separately administered vaccines. Pentaceltrade mark (Sanofi Pasteur Ltd., Toronto, Canada), a combination vaccine first licensed in Canada and subsequently in other countries, is immunogenic against diphtheria, tetanus, pertussis, polio and Haemophilus influenzae type b when administered at 2, 4, 6 and 15-18 months of age. In published studies, the safety, immunogenicity and effectiveness of this combination vaccine were comparable with those of separately administered vaccines, with the advantage of a simplified dosing schedule. PMID- 16372876 TI - New vaccines against enteric bacteria for children in less developed countries. AB - Diarrheal diseases represent a major threat to infant survival in less developed countries. A real opportunity now exists to help alleviate this problem through the development of safe and effective multicomponent whole-bacterial cell vaccine(s) against enterotoxigenic Escherichia coli and Shigella, two important pathogens for which no licensed vaccine exists. What is preventing realization of this achievement is a lack of focus on the unique needs of children in less developed countries, along with committed and sufficient funding directed toward this goal. Live-attenuated and inactivated whole-cell vaccine candidates, some of which have languished too long, are available for testing, which, if performed in a coordinated fashion, can answer key unresolved issues concerning mucosal vaccination against enteric diseases. These candidate vaccines potentially provide a relatively simple intervention which could, if implemented, reduce the impact of these diseases upon the life and productivity of children. PMID- 16372877 TI - Maternal immunization to protect the mother and neonate. AB - The concept of maternal immunization to protect the mother against vaccine preventable disease and the neonate against illness for the first 3-6 months of life is one that is simple, straightforward and safe. Neonatal immunization is largely unsuccessful due to immaturity of the infant's immune system. Therefore, appropriate maternal immunization and passive transferred antibodies to the fetus can protect the neonate until infant vaccination is more efficacious. PMID- 16372878 TI - Prevention of Haemophilus influenzae type b disease: past success and future challenges. AB - Haemophilus influenzae type b (Hib) is responsible for significant morbidity and mortality worldwide, particularly in children under 5 years of age. In countries where the Hib conjugate vaccine is not routinely used, Hib is a leading cause of childhood pneumonia and meningitis. Routine use of the Hib conjugate vaccines has resulted in a remarkable decline in Hib disease in developed and developing countries. However, Hib conjugate vaccines are not routinely available in most developing countries, many of which have high burdens of Hib disease. This review outlines the pathogenesis and epidemiology of Hib disease, and the various options for prevention. PMID- 16372879 TI - Towards a therapeutic breast cancer vaccine: the next steps. AB - The genomic revolution has advanced our understanding of breast cancer biology and the molecular basis of antitumor immunity. Trastuzumab, the first monoclonal antibody for breast cancer, is now a routine part of clinical care. Breast cancer vaccines may be more effective by actively recruiting both humoral and cellular immunity to the therapeutic effort. However, immunization alone is unlikely to have significant activity against established breast cancers, where it is limited by potent mechanisms of immune tolerance and the immunobiology of breast cancer itself. The next generation of clinical studies should integrate breast cancer vaccines with standard breast cancer drugs or novel immunotherapeutics in strategic doses and schedules that abrogate immune tolerance and groom the tumor microenvironment for a productive immune response. PMID- 16372880 TI - Vaccine development for non-typeable Haemophilus influenzae and Moraxella catarrhalis: progress and challenges. AB - An urgent need exists for vaccines to prevent infections caused by nontypeable Haemophilus influenzae and Moraxella catarrhalis. These bacteria cause otitis media in children, a clinical problem associated with enormous morbidity and cost. H. influenzae and M. catarrhalis also cause lower respiratory tract infections in adults with chronic lung disease. Infections in this clinical setting are associated with disability and death. Recent progress in identifying potential vaccine antigens in both bacteria raises great promise in developing effective vaccines. This paper reviews the key issues in vaccine development for H. influenzae and M. catarrhalis, including areas where progress has been stalled, and proposes areas that deserve investigation in the next 5 years. PMID- 16372881 TI - Ensuring the quality, potency and safety of vaccines during preclinical development. AB - There is an abundance of vaccines currently in development, with most of them exploring novel mechanisms, adjuvants and/or delivery systems not only for traditional prophylactic use, but also for therapeutic uses. As vaccines are generally administered to healthy individuals, ensuring their quality, potency and safety becomes crucial, especially prior to evaluation in humans. To ensure these key attributes, vaccine developers need to incorporate them as early in the development program as possible, starting in basic research and continuing through preclinical, clinical and postmarketing development. Fortunately for vaccine developers, ample guidance is available from various regulatory agencies to enlighten the long and arduous path of vaccine development. This review will highlight these regulatory expectations, and provide some clarity as to why they are in place. PMID- 16372882 TI - Current status and future prospects for a vaccine against American trypanosomiasis. AB - The clinically relevant pathognomonic consequences of human infection by Trypanosoma cruzi are dilation and hypertrophy of the left ventricle walls and thinning of the apex. The major complications and debilitating evolutionary outcomes of chronic infection include ventricular fibrillation, thromboembolism and congestive heart failure. American trypanosomiasis (Chagas disease) poses serious public healthcare and budgetary concerns. The currently available drugs, although effective against acute infection, are highly toxic and ineffective in arresting or attenuating clinical disease symptoms in chronic patients. The development of an efficacious prophylactic vaccine faces many challenges, and progress is slow, despite several years of effort. Studies in animal models and human patients have revealed the pathogenic mechanisms during disease progression, pathology of disease and features of protective immunity. Accordingly, several antigens, antigen-delivery vehicles and adjuvants have been tested in animal models, and some efforts have been successful in controlling infection and disease. This review will summarize the accumulated knowledge about the parasite and disease, as well as pathogenesis and protective immunity. The authors will discuss the efforts to date, and the challenges faced in achieving an efficient prophylactic vaccine against human American trypanosomiasis, and present the future perspectives. PMID- 16372883 TI - Peptide-based immunotherapy: a novel strategy for allergic disease. AB - The T-cell component of the antigen-specific immune response is the target of various novel interventions to modify chronic immunologic disorders, such as allergic diseases. Recent clinical trials have evaluated the safety and efficacy of therapeutic vaccines consisting of short, synthetic, allergen-derived peptides, corresponding to T-cell epitopes from the eliciting antigen. The main advantage of such an approach is the reduction in systemic, immunoglobulin E mediated adverse events compared with existing whole allergen immunotherapy, often referred to as 'allergy shots'. T-cell peptide epitopes, although capable of inducing immunologic tolerance, are short linear structures that have reduced ability to cross-link mast cell- and basophil-bound immunoglobulin E. The precise mechanism of tolerance induction remains incompletely defined. However, recent data indicate that peptide therapy induces/expands a population of antigen specific regulatory T-cells. A novel form of treatment combining efficacy with a substantially decreased occurrence of adverse events is likely to have a major impact on the management and prevalence of allergic diseases. Furthermore, the principles of epitope-specific therapy hold promise for the development of therapeutic vaccines for the treatment of autoimmune diseases. PMID- 16372884 TI - Vaccines for bovine tuberculosis: current views and future prospects. AB - Bovine tuberculosis, caused by Mycobacterium bovis, is rapidly increasing in cattle herds in developed countries such as the UK, New Zealand and the USA. In addition, persistence of M. bovis in other parts of the world may account for up to 10% of cases of human tuberculosis. Thus, a rise in the number of M. bovis infections poses an increased human health risk and is also a major economic problem. In the UK, the incidence of bovine tuberculosis continues to rise despite the use of a skin test and slaughter control policy, highlighting the need for an effective vaccination strategy to control the spread of disease. The only vaccine currently available for human, (and bovine), tuberculosis is Bacillus Calmette-Guerin, which is known to have variable efficacy for both species. In this article, the authors discuss potential strategies by which Bacillus Calmette-Guerin vaccination may be improved to allow highly efficacious vaccination of cattle. These strategies are also highly applicable to the fight against tuberculosis in humans. PMID- 16372885 TI - Evolving perception on the benefits of vaccination as a foot and mouth disease control policy: contributions of South America. AB - Within the past decade, changes in perceptions on the benefits of vaccination as an appropriate tool to achieve complete foot and mouth disease eradication have become evident. The former negative view was derived from misconceptions, resulting mainly from the belief that vaccines are not entirely effective and that vaccination masks asymptomatic viral circulation. The advent in the 1990s of vaccination policies implemented within a strategic eradication plan in South America, and during recurrence of the disease in disease-free regions contributed towards generating more reliable and visible outcomes of vaccination programs, paving the way towards a new perception. Particularly relevant was the development and application of novel serodiagnostic approaches to assess silent viral circulation, irrespective of vaccination. The use in South America of vaccination allied to serosurveys to accompany viral clarification during eradication campaigns and after emergencies clearly established the importance of this control tool to stop the spread of viral infection. This alliance gave input to break many myths associated with the use of vaccines, including the belief that immunized carrier animals pose an epidemiologic risk. This experience launched new concepts that supported the internationally recognized status of foot and mouth disease-free regions with vaccination and the 'vaccination to live' policy as an alternative to 'stamping out'. PMID- 16372886 TI - Essential role of vaccines in brucellosis control and eradication programs for livestock. AB - Brucellosis, in particular infections with Brucella abortus, Brucella melitensis or Brucella suis, remains a significant human health threat in many areas of the world. The persistence of pathogenic Brucella spp. in domestic livestock or free ranging wildlife remains unresolved, despite decades of regulatory efforts worldwide. Although vaccination is probably the most economic control measure, administration of currently available vaccines alone is not sufficient for elimination of brucellosis in any host species. Complacency in brucellosis control programs usually results in failure, or at best, limited reductions in disease prevalence or incidence of human infections. New brucellosis vaccines with high efficacy and safety are needed that address the diversity in host species and can be more widely applied under field conditions. Development of safer and more efficacious vaccines alone, or combined with enhancements or increased emphasis on other regulatory program components, could have tremendous impact on reducing the worldwide prevalence of brucellosis and the associated zoonotic infections. PMID- 16372888 TI - Oral health during pregnancy: current research. AB - This report describes recent efforts by the Centers for Disease Control and Prevention (CDC), Division of Oral Health, to understand more fully women's knowledge and attitudes regarding oral health and dental visits during pregnancy. Using data from the CDC Pregnancy Risk Assessment Monitoring System (PRAMS), investigators are conducting both quantitative and qualitative research on these issues. PRAMS is an ongoing state-based and population-based surveillance survey of women's attitudes, experiences, and behaviors before, during, and after pregnancy. Findings have shown that most mothers did not make a dental visit during pregnancy, and of those who reported having oral problems, one-half did not seek care. Preliminary analysis of qualitative results shows that some women may believe that poor oral health status during pregnancy is normal; also, they may fear certain aspects of dental care during pregnancy. For example, some women may believe that they or their fetus could be harmed by treatment. If pregnancy modifies perceptions of oral health and dental care in women, it may contribute to women's avoidance of dental treatment while pregnant. Therefore, researchers and health program planners should give increased attention to the oral health needs and behaviors of pregnant women. PMID- 16372889 TI - Toward optimal health: Scott Grundy, M.D., Ph.D. discusses metabolic syndrome ( interviewed by Jodi R. Godfrey). PMID- 16372890 TI - Interpreting studies of nutritional prevention: a perspective using calcium as a model. PMID- 16372891 TI - Impact of street drug use, HIV infection, and highly active antiretroviral therapy on reproductive hormones in middle-aged women. AB - OBJECTIVE: To assess the impact of street drug use and HIV infection on reproductive hormones in 82 women aged 28?56 and 15 HIV-uninfected, regularly cycling premenopausal historical controls. METHODS: Prospective, pilot cohort study. Baseline blood samples were assayed for follicle stimulating hormone (FSH), human chorionic gonadotropin (hCG), prolactin (PRL), thyroid stimulating hormone (TSH), and estradiol (E(2)). Menopausal status was defined as premenopause: age<40, not amenorrheic; perimenopause: age>40, not amenorrheic; menopause: age>40, with> or =12 months' amenorrhea. Kruskal-Wallis testing was used to compare groups of women sorted by menopausal status and separated by drug use and HIV serostatus. Controls were regularly cycling premenopausal women. RESULTS: Thirty-eight of the 82 women (46%) reported substance abuse, and 47 of the 82 (57%) were HIV infected. TSH did not differ by HIV serostatus or drug use. PRL was elevated in drug users compared with nonusers and healthy volunteers (10.3, 5.9 vs. 6.2 ng/ml, respectively, p = 0.002), with no effect of HIV serostatus. FSH was reduced in each menstrual category related to drug use and in postmenopausal women associated with positive HIV serostatus. Highly active antiretroviral therapy (HAART) use was not related to PRL or E(2) but was associated with higher FSH. FSH was greater in cohort participants compared with controls. CONCLUSIONS: Drug use, not HIV, relates to increased PRL. Both drug use and HIV infection are associated with decreased FSH. Women in this socioeconomic stratum at high risk for HIV may be at risk for early menopause. Increased PRL may falsely reduce FSH, necessitating a more careful hormonal characterization of menopausal status in this sample of women. PMID- 16372892 TI - Urinary incontinence: proper assessment and available treatment options. AB - Urinary incontinence is a common condition that imposes significant burdens on the health and well-being of the patient. Urinary incontinence is underreported because people who suffer from the disorder are often reluctant to discuss it with healthcare professionals. It is undertreated because of misperceptions regarding its etiology and amenability to therapy. Failure to distinguish among the different types of urinary incontinence can hinder optimal treatment. This summary paper reviews the types of urinary incontinence, its prevalence, its impact on medical costs and patient quality of life, and approaches to diagnosis and treatment. PMID- 16372894 TI - Postpartum depression assessments at well-baby visits: screening feasibility, prevalence, and risk factors. AB - BACKGROUND: Postpartum depression (PPD) is a disorder with broad public health implications and consequences that impact almost every aspect of child development. METHODS: In this pilot study, study participants were 96 women who brought their babies to the University of Arizona Pediatrics Clinic for their 8 week well-baby visit. Participants completed a packet that consisted of questions about demographics, potential correlates of PPD, and the Edinburgh Postpartum Depression Scale (EPDS). English and Spanish versions were available. RESULTS: Of a total of 172 women who brought their babies in for their 8-week well-baby visit, 96 women completed the packets, for an overall response rate of 56.9%. Observed EPDS scores ranged from 0 to 18, with a mean of 5.44 and a standard deviation (SD) of 4.83. Using the cutoff of EPDS > or = 12, 14.6% of participants were likely suffering from clinically significant depression. Higher EPDS scores and also categorical depression classification were statistically associated with reported smoking and a family history of mental health problems. CONCLUSIONS: We conclude that screening for mothers at well-baby visits is feasible and that the data collected are of sufficient quality to identify reliable predictors even with small sample sizes. PMID- 16372893 TI - Do long-term HDL-C declines associated with a first birth vary by apo E phenotype? The Coronary Artery Risk Development in Young Adults (CARDIA) study. AB - BACKGROUND: High-density lipoprotein cholesterol (HDL-C) levels in premenopausal and postmenopausal women are differentially affected by exogenous sex hormones depending on their apolipoprotein E (apo E) genotype. Because endogenous sex hormones markedly increase during pregnancy, we examined whether HDL-C declines after a first birth varied by apo E polymorphisms. METHODS: In 1147 nulliparas (416 black, 731 white), fasting blood samples (nonpregnant) were drawn at baseline and at follow-up years 5, 7, and 10. Time-dependent pregnancy groups included 0 pregnancies (P0), 1+ short pregnancy (P1+), 1 birth (B1), 2 or more births (B2+). ApoE groups by alleles identified with a phenotype method included E4 (4/3 and 4/4), E3 (3/3), and E2 (2/2 and 3/2). Differences in adjusted mean HDL-C changes among pregnancy groups and ApoE groups were examined using repeated measures multiple linear regression. RESULTS: HDL-C declines associated with parity (one or more births) depended on ApoE group (ApoE*Pregnancy Interaction; p < 0.002). For B1 and B2+ vs. P0, HDL-C declines were -2.4 to -2.7 mg/dl in E4 and -3.4 to -4.1 mg/dl in E3. In E2, HDL-C declines were -6.6 mg/dl for one birth, and -11.5 mg/dl for two or more births, each relative to the 0 pregnancies (P0) group (linear trend, p < 0.001). CONCLUSIONS: The degree to which childbearing adversely affects long-term HDL-C declines varies by apo E phenotype, based on a method that accurately classifies genotype. Our findings show that 2/2 and 3/2 genotypes are associated with larger parity-related HDL-C declines than 3/3, 4/3, and 4/4 genotypes. PMID- 16372895 TI - Evaluation of web-based osteoporosis educational materials. AB - BACKGROUND: Many women are unaware of risk factors for and consequences of osteoporosis. Hence, patient education is an essential step in preventing and managing osteoporosis. Unfortunately, numerous studies have demonstrated a mismatch between the reading difficulty of typical patient education materials and the reading ability of many American adults. METHODS: We examined the readability and quality of web-based information on osteoporosis using the Suitability Assessment of Materials (SAM) and DISCERN instruments. The three most widely used Internet search engines, Google, Yahoo, and MSN, were selected based on popularity according to Nielsen/Net Ratings. The search term osteoporosis was entered to generate the first 30 websites listed by each of the three search engines. Several websites appeared on all three search engines; we identified 27 unique websites. RESULTS: Overall, 51.9% of materials were rated by the SAM as not suitable. Most materials scored poorly for their reading level (82.6% were rated not suitable), with an average reading level at grade 11.5 +/- 2.8. The mean DISCERN score for overall description of treatment options was 35.7 +/- 18.0, indicating inadequate quality. Most materials had low quality in a number of indicators, including accuracy and biased presentation of information. CONCLUSIONS: Web-based osteoporosis information is written above the reading ability of most American adults, and much of it lacks adequate quality. PMID- 16372896 TI - An exploratory study of mother-daughter physicians: an intergenerational comparison of professional and personal experiences. AB - OBJECTIVES: Physician daughters of physician mothers may experience unique advantages in their career development and in combining career and family. The objective of this exploratory study, the first on mother-daughter physicians, was to compare the professional and personal characteristics of physician mothers and their physician daughters. METHODS: Two hundred fourteen families with at least one mother-daughter physician pair were identified through a nationwide search; 84% of the mothers and 87% of the daughters contacted returned a 56-item questionnaire. Data were analyzed to compare professional and personal characteristics of the subset of mothers and daughters who had both responded (n = 136 pairs). RESULTS: Compared to their physician mothers, physician daughters were less satisfied with their career, less likely to feel in control of their work environment, more likely to report moderate or severe daily stress at work, and less likely to want their children to become physicians or to want to become physicians again if they were to relieve their lives. They were also less likely to want to change specialties. Physician daughters were more likely than their physician mothers to be 30 years or older when they had their first child, to have taken parental leave, and to have a spouse who took parental leave and less likely to experience their caregiving as a hindrance to their professional work and advancement. CONCLUSIONS: Despite the availability of a physician mother role model, physician daughters reported less career satisfaction and more work related stress than their physician mothers. These findings warrant further exploration and support the need for new work-family paradigms in medicine. PMID- 16372898 TI - Functional properties of ryanodine receptors carrying three amino acid substitutions identified in patients affected by multi-minicore disease and central core disease, expressed in immortalized lymphocytes. AB - More than 80 mutations in the skeletal muscle ryanodine receptor gene have been found to be associated with autosomal dominant forms of malignant hyperthermia and central core disease, and with recessive forms of multi-minicore disease. Studies on the functional effects of pathogenic dominant mutations have shown that they mostly affect intracellular Ca2+ homoeostasis, either by rendering the channel hypersensitive to activation (malignant hyperthermia) or by altering the amount of Ca2+ released subsequent to physiological or pharmacological activation (central core disease). In the present paper, we show, for the first time, data on the functional effect of two recently identified recessive ryanodine receptor 1 amino acid substitutions, P3527S and V4849I, as well as that of R999H, another substitution that was identified in two siblings that were affected by multi minicore disease. We studied the intracellular Ca2+ homoeostasis of EBV (Epstein Barr virus)-transformed lymphoblastoid cells from the affected patients, their healthy relatives and control individuals. Our results show that the P3527S substitution in the homozygous state affected the amount of Ca2+ released after pharmacological activation with 4-chloro-m-cresol and caffeine, but did not affect the size of the thapsigargin-sensitive Ca2+ stores. The other substitutions had no effect on either the size of the intracellular Ca2+ stores, or on the amount of Ca2+ released after ryanodine receptor activation; however, both the P3527S and V4849I substitutions had a small but significant effect on the resting Ca2+ concentration. PMID- 16372899 TI - Myosin heavy-chain kinase A from Dictyostelium possesses a novel actin-binding domain that cross-links actin filaments. AB - Myosin heavy-chain kinase A (MHCK A) catalyses the disassembly of myosin II filaments in Dictyostelium cells via myosin II heavy-chain phosphorylation. MHCK A possesses a 'coiled-coil'-enriched domain that mediates the oligomerization, cellular localization and actin-binding activities of the kinase. F-actin (filamentous actin) binding by the coiled-coil domain leads to a 40-fold increase in MHCK A activity. In the present study we examined the actin-binding characteristics of the coiled-coil domain as a means of identifying mechanisms by which MHCK A-mediated disassembly of myosin II filaments can be regulated in the cell. Co-sedimentation assays revealed that the coiled-coil domain of MHCK A binds co-operatively to F-actin with an apparent K(D) of approx. 0.5 muM and a stoichiometry of approx. 5:1 [actin/C(1-498)]. Further analyses indicate that the coiled-coil domain binds along the length of the actin filament and possesses at least two actin-binding regions. Quite surprisingly, we found that the coiled coil domain cross-links actin filaments into bundles, indicating that MHCK A can affect the cytoskeleton in two important ways: (1) by driving myosin II-filament disassembly via myosin II heavy-chain phosphorylation, and (2) by cross linking/bundling actin filaments. This discovery, along with other supporting data, suggests a model in which MHCK A-mediated bundling of actin filaments plays a central role in the recruitment and activation of the kinase at specific sites in the cell. Ultimately this provides a means for achieving the robust and highly localized disruption of myosin II filaments that facilitates polarized changes in cell shape during processes such as chemotaxis, cytokinesis and multicellular development. PMID- 16372900 TI - Structure and stability of 2S albumin-type peanut allergens: implications for the severity of peanut allergic reactions. AB - Resistance to proteolytic enzymes and heat is thought to be a prerequisite property of food allergens. Allergens from peanut (Arachis hypogaea) are the most frequent cause of fatal food allergic reactions. The allergenic 2S albumin Ara h 2 and the homologous minor allergen Ara h 6 were studied at the molecular level with regard to allergenic potency of native and protease-treated allergen. A high resolution solution structure of the protease-resistant core of Ara h 6 was determined by NMR spectroscopy, and homology modelling was applied to generate an Ara h 2 structure. Ara h 2 appeared to be the more potent allergen, even though the two peanut allergens share substantial cross-reactivity. Both allergens contain cores that are highly resistant to proteolytic digestion and to temperatures of up to 100 degrees C. Even though IgE antibody-binding capacity was reduced by protease treatment, the mediator release from a functional equivalent of a mast cell or basophil, the humanized RBL (rat basophilic leukaemia) cell, demonstrated that this reduction in IgE antibody-binding capacity does not necessarily translate into reduced allergenic potency. Native Ara h 2 and Ara h 6 have virtually identical allergenic potency as compared with the allergens that were treated with digestive enzymes. The folds of the allergenic cores are virtually identical with each other and with the fold of the corresponding regions in the undigested proteins. The extreme immunological stability of the core structures of Ara h 2 and Ara h 6 provides an explanation for the persistence of the allergenic potency even after food processing. PMID- 16372901 TI - Avoiding inconsistencies over time and tracking difficulties in Applied Biosystems AB1700/Panther probe-to-gene annotations. AB - BACKGROUND: Significant inconsistencies between probe-to-gene annotations between different releases of probe set identifiers by commercial microarray platform solutions have been reported. Such inconsistencies lead to misleading or ambiguous interpretation of published gene expression results. RESULTS: We report here similar inconsistencies in the probe-to-gene annotation of Applied Biosystems AB1700 data, demonstrating that this is not an isolated concern. Moreover, the online information source PANTHER does not provide information required to track such inconsistencies, hence, even correctly annotated datasets, when resubmitted after PANTHER was updated to a new probe-to-gene annotation release, will generate differing results without any feedback on the origin of the change. CONCLUSION: The importance of unequivocal annotation of microarray experiments can not be underestimated. Inconsistencies greatly diminish the usefulness of the technology. Novel methods in the analysis of transcriptome profiles often rely on large disparate datasets stemming from multiple sources. The predictive and analytic power of such approaches rapidly diminishes if only least-common subsets can be used for analysis. We present here the information that needs to be provided together with the raw AB1700 data, and the information required together with the biologic interpretation of such data to avoid inconsistencies and tracking difficulties. PMID- 16372902 TI - An automated, broad-based, near real-time public health surveillance system using presentations to hospital Emergency Departments in New South Wales, Australia. AB - BACKGROUND: In a climate of concern over bioterrorism threats and emergent diseases, public health authorities are trialling more timely surveillance systems. The 2003 Rugby World Cup (RWC) provided an opportunity to test the viability of a near real-time syndromic surveillance system in metropolitan Sydney, Australia. We describe the development and early results of this largely automated system that used data routinely collected in Emergency Departments (EDs). METHODS: Twelve of 49 EDs in the Sydney metropolitan area automatically transmitted surveillance data from their existing information systems to a central database in near real-time. Information captured for each ED visit included patient demographic details, presenting problem and nursing assessment entered as free-text at triage time, physician-assigned provisional diagnosis codes, and status at departure from the ED. Both diagnoses from the EDs and triage text were used to assign syndrome categories. The text information was automatically classified into one or more of 26 syndrome categories using automated "naive Bayes" text categorisation techniques. Automated processes were used to analyse both diagnosis and free text-based syndrome data and to produce web-based statistical summaries for daily review. An adjusted cumulative sum (cusum) was used to assess the statistical significance of trends. RESULTS: During the RWC the system did not identify any major public health threats associated with the tournament, mass gatherings or the influx of visitors. This was consistent with evidence from other sources, although two known outbreaks were already in progress before the tournament. Limited baseline in early monitoring prevented the system from automatically identifying these ongoing outbreaks. Data capture was invisible to clinical staff in EDs and did not add to their workload. CONCLUSION: We have demonstrated the feasibility and potential utility of syndromic surveillance using routinely collected data from ED information systems. Key features of our system are its nil impact on clinical staff, and its use of statistical methods to assign syndrome categories based on clinical free text information. The system is ongoing, and has expanded to cover 30 EDs. Results of formal evaluations of both the technical efficiency and the public health impacts of the system will be described subsequently. PMID- 16372903 TI - Understanding non-compliance to colorectal cancer screening: a case control study, nested in a randomised trial [ISRCTN83029072]. AB - BACKGROUND: The major limit to colorectal cancer screening effectiveness is often low compliance. We studied the reasons for non compliance and determinants of compliance to faecal occult blood tests in Lazio, Italy. METHODS: This is a case control study nested within a trial that tested the effect of type of test and provider on colorectal cancer screening compliance. Non compliant trial subjects were classified as cases, and compliant subjects were classified as controls. We sampled 600 cases and 600 controls matched by their general practitioner, half were invited for screening at the hospital, and the other half directly at their general practitioner's office. Cases and controls answered questions on: distance from test provider, logistical problems, perception of colorectal cancer risk, confidence in screening efficacy, fear of results, presence of colorectal cancer in the family, and gastrointestinal symptoms. RESULTS: About 31% of cases never received the letter offering free screening, and 17% of the sampled population had already been screened. The first reported reason for non-compliance was "lack of time" (30%); the major determinant of compliance was the distance from the test provider: odds ratio > 30 minutes vs < 15 minutes 0.3 (95% CI = 0.2-0.7). The odds ratio for lack of time was 0.16 (95% IC 0.1-0.26). The effect was stronger if the hospital (0.03 95% CI = 0.01-0.1) rather than the general practitioner (0.3 95% CI = 0.2-0.6) was the provider. Twenty-two percent of controls were accompanied by someone to the test. CONCLUSION: To increase compliance, screening programmes must involve test providers who are geographically close to the target population. PMID- 16372904 TI - Mediterranean diet or extended fasting's influence on changing the intestinal microflora, immunoglobulin A secretion and clinical outcome in patients with rheumatoid arthritis and fibromyalgia: an observational study. AB - BACKGROUND: Alterations in the intestinal bacterial flora are believed to be contributing factors to many chronic inflammatory and degenerative diseases including rheumatic diseases. While microbiological fecal culture analysis is now increasingly used, little is known about the relationship of changes in intestinal flora, dietary patterns and clinical outcome in specific diseases. To clarify the role of microbiological culture analysis we aimed to evaluate whether in patients with rheumatoid arthritis (RA) or fibromyalgia (FM) a Mediterranean diet or an 8-day fasting period are associated with changes in fecal flora and whether changes in fecal flora are associated with clinical outcome. METHODS: During a two-months-period 51 consecutive patients from an Integrative Medicine hospital department with an established diagnosis of RA (n = 16) or FM (n = 35) were included in the study. According to predefined clinical criteria and the subjects' choice the patients received a mostly vegetarian Mediterranean diet (n = 21; mean age 50.9 +/-13.3 y) or participated in an intermittent modified 8-day fasting therapy (n = 30; mean age 53.7 +/- 9.4 y). Quantitative aerob and anaerob bacterial flora, stool pH and concentrations of secretory immunoglobulin A (sIgA) were analysed from stool samples at the beginning, at the end of the 2-week hospital stay and at a 3-months follow-up. Clinical outcome was assessed with the DAS 28 for RA patients and with a disease severity rating scale in FM patients. RESULTS: We found no significant changes in the fecal bacterial counts following the two dietary interventions within and between groups, nor were significant differences found in the analysis of sIgA and stool ph. Clinical improvement at the end of the hospital stay tended to be greater in fasting vs. non-fasting patients with RA (p = 0.09). Clinical outcome was not related to alterations in the intestinal flora. CONCLUSION: Neither Mediterranean diet nor fasting treatments affect the microbiologically assessed intestinal flora and sIgA levels in patients with RA and FM. The impact of dietary interventions on the human intestinal flora and the role of the fecal flora in rheumatic diseases have to be clarified with newer molecular analysis techniques. The potential benefit of fasting treatment in RA and FM should be further tested in randomised trials. PMID- 16372905 TI - The feasibility of Whole Body Vibration in institutionalised elderly persons and its influence on muscle performance, balance and mobility: a randomised controlled trial [ISRCTN62535013]. AB - BACKGROUND: Fatigue or lack of interest can reduce the feasibility of intensive physical exercise in nursing home residents. Low-volume exercise interventions with similar training effects might be an alternative. The aim of this randomised controlled trial was to investigate the feasibility of Whole Body Vibration (WBV) in institutionalised elderly, and its impact on functional capacity and muscle performance. METHODS: Twenty-four nursing home residents (15 female, 9 male; mean age 77.5 +/- 11.0 years) were randomised (stratification for age, gender and ADL category) to 6 weeks static WBV exercise (WBV+, N = 13) or control (only static exercise; N = 11). Outcome measures were exercise compliance, timed up-and-go, Tinetti-test, back scratch, chair sit-and-reach, handgrip strength and linear isokinetic leg extension. RESULTS: At baseline, WBV+ and control groups were similar for all outcome variables. Twenty-one participants completed the program and attended respectively 96% and 86% of the exercise sessions for the WBV+ and control groups. Training-induced changes in timed up-and-go and Tinetti-test were better for WBV+ compared to control (p = 0.029 for timed up-and-go, p = 0.001 and p = 0.002 for Tinetti body balance and total score respectively). In an alternative analysis (Worst Rank Score & Last Observation Carried Forward) the differences in change remained significant on the Tinetti body balance and total score. No other significant differences in change between both groups were observed. CONCLUSION: In nursing home residents with limited functional dependency, six weeks static WBV exercise is feasible, and is beneficial for balance and mobility. The supplementary benefit of WBV on muscle performance compared to classic exercise remains to be explored further. PMID- 16372906 TI - Failure to confirm influence of methyltetrahydrofolate reductase (MTHFR) polymorphisms on age at onset of Huntington disease. AB - BACKGROUND: Huntington disease (HD) is a fully penetrant, autosomal dominantly inherited disorder associated with abnormal expansions of a stretch of perfect CAG repeats in the 5' part of the IT15 gene. The number of repeat units is highly predictive for the age at onset (AO) of the disorder. But AO is only modestly correlated with repeat length when intermediate HD expansions are considered. Recently, suggestive association has been reported between a single nucleotide polymorphism (SNP; rs1801131, also known as A1298C) in the methyltetrahydrofolate reductase (MTHFR) gene and AO of HD. 5,10-MTHFR is a key enzyme in the folate metabolism, diverting metabolites toward methylation reactions or nucleotide synthesis. Using part of a previously established study cohort plus additional patients and appropriate statistical methods, we reinvestigated two polymorphisms in the MTHFR gene, C677T and A1298C, as well as their association with AO in 167 HD patients. RESULTS: There was no statistically significant impact on AO for HD patients, neither of MTHFR SNPs nor of the combinations thereof. CONCLUSION: Contrary to previously described evidence the A1298C polymorphism in the MTHFR gene does not appear to modulate AO of HD patients. PMID- 16372907 TI - MMP-9, TIMP-1 and inflammatory cells in sputum from COPD patients during exacerbation. AB - BACKGROUND: Irreversible airflow obstruction in Chronic Obstructive Pulmonary Disease (COPD) is thought to result from airway remodelling associated with aberrant inflammation. Patients who experience frequent episodes of acute deterioration in symptoms and lung function, termed exacerbations, experience a faster decline in their lung function, and thus over time greater disease severity However the mechanisms by which these episodes may contribute to decreased lung function are poorly understood. This study has prospectively examined changes in sputum levels of inflammatory cells, MMP-9 and TIMP-1 during exacerbations comparing with paired samples taken prior to exacerbation. METHODS: Nineteen COPD patients ((median, [IQR]) age 69 [63 to 74], forced expiratory volume in one second (FEV1) 1.0 [0.9 to 1.2], FEV1% predicted 37.6 [27.3 to 46.2]) provided sputa at exacerbation. Of these, 12 were paired with a samples collected when the patient was stable, a median 4 months [2 to 8 months] beforehand. RESULTS: MMP-9 levels increased from 10.5 microg/g [1.2 to 21.1] prior to exacerbation to 17.1 microg/g [9.3 to 48.7] during exacerbation (P < 0.01). TIMP-1 levels decreased from 3.5 microg/g [0.6 to 7.8] to 1.5 microg/g [0.3 to 4.9] (P = 0.16). MMP-9/TIMP-1 Molar ratio significantly increased from 0.6 [0.2 to 1.1] to 3.6 [2.0 to 25.3] (P < 0.05). Neutrophil, eosinophil and lymphocyte counts all showed significant increase during exacerbation compared to before (P < 0.05). Macrophage numbers remained level. MMP-9 levels during exacerbation showed highly significant correlation with both neutrophil and lymphocyte counts (Rho = 0.7, P < 0.01). CONCLUSION: During exacerbation, increased inflammatory burden coincides with an imbalance of the proteinase MMP-9 and its cognate inhibitor TIMP-1. This may suggest a pathway connecting frequent exacerbations with lung function decline. PMID- 16372908 TI - Pain management procedures used by dental and maxillofacial surgeons: an investigation with special regard to odontalgia. AB - BACKGROUND: Little is known about the procedures used by German dental and maxillofacial surgeons treating patients suffering from chronic orofacial pain (COP). This study aimed to evaluate the ambulatory management of COP. METHODS: Using a standardized questionnaire we collected data of dental and maxillofacial surgeons treating patients with COP. Therapists described variables as patients' demographics, chronic pain disorders and their aetiologies, own diagnostic and treatment principles during a period of 3 months. RESULTS: Although only 13.5% of the 520 addressed therapists returned completely evaluable questionnaires, 985 patients with COP could be identified. An orofacial pain syndrome named atypical odontalgia (17.0 %) was frequent. Although those patients revealed signs of chronification, pain therapists were rarely involved (12.5%). For assessing pain the use of Analogue Scales (7%) or interventional diagnostics (4.6%) was uncommon. Despite the fact that surgical procedures are cofactors of COP therapists preferred further surgery (41.9%) and neglected the prescription of analgesics (15.7%). However, most therapists self-evaluated the efficacy of their pain management as good (69.7 %). CONCLUSION: Often ambulatory dental and maxillofacial surgeons do not follow guidelines for COP management despite a high prevalence of severe orofacial pain syndromes. PMID- 16372909 TI - Cytochrome P450 aromatase expression in human seminoma. AB - BACKGROUND: The enzyme cytochrome P450 aromatase, catalysing the conversion of androgens into estrogens, has been detected in normal human testicular cells suggesting a physiological role of local estrogen biosynthesis on spermatogenesis control. Estrogens, regulating cell growth and apoptosis, can also be involved in tumorigenesis process, but the possible link between estrogens and testicular neoplastic process is, up to now, scarcely known. This study examined aromatase expression in human seminoma, which is the most common germ cell tumour of the testis. METHODS: The tumour-bearing testes were obtained from 20 patients with classic seminoma undergoing to therapeutic orchidectomy. Paraffin embedded tissues were processed for immunohistochemistry using a mouse monoclonal antibody generated against human placental cytochrome P450 arom, as primary antibody, and a biotinylated goat-anti-mouse IgG, as secondary antibody. Furthermore, Western blot analysis of seminoma extracts was carried out. RESULTS: Intense P450 arom immunoreactivity was observed in the seminoma cells and Western blot analysis confirmed the immunodetection. A strong immunostaining was also detected in cells of intratubular germ cell neoplasia (IGCN), adjacent to seminoma. CONCLUSION: The present study demonstrated, for the first time in human, aromatase expression in neoplastic cells of seminoma suggesting a relation between local estrogen biosynthesis and germ cell tumorigenesis. The P450 arom immunolocalization in the cells of IGCN, representing the common precursor of most germ cell tumors, seems to support these findings. PMID- 16372910 TI - Risk factors associated with mental illness in Oyo State, Nigeria: a community based study. AB - BACKGROUND: The main objective of this study was to determine the prevalence and factors associated with mental illness in Oyo State at community level using the general health questionnaire as a screening tool. METHOD: This cross-sectional, community- based survey was carried out among adults in three randomly selected LGAs using multi-stage sampling technique. RESULTS: A total of 1105 respondents were assessed in all. The overall prevalence of psychiatric morbidity in Oyo state Nigeria was found to be 21.9%, (18.4% in the urban areas and 28.4% in the rural areas, p = 0.005). Young age < or = 19 yrs (X2 = 20.41, p = 0.00013), Unemployment (X2 = 11.86 p = 0.0005), living condition below average (X2 = 12.21, p = 0.00047), physical health (X2 = 6.07, p = 0.014), and large family size (X2 = 14.09 p = 0.00017) were associated with increase risk for psychiatric morbidity. Following logistic regression analysis, Unemployment (C.I = 1.18-3.70, OR -2.1) and living conditions perceived to be above average (C.I = 1.99-5.50, OR-3.3) were significant predictors of mental illness while family size less than 6 (C.I = 0.86-0.97, OR-0.91) was protective. CONCLUSION: The teenagers and the rural populations are in greater need of mental health promotional services. Family planning should be made freely available in order to reduce the family size and hence incidence of mental illness in the African population. PMID- 16372911 TI - Please understand when I cry out in pain: women's accounts of maternity services during labour and delivery in Ghana. AB - BACKGROUND: This study was undertaken to investigate women's accounts of interactions with health care providers during labour and delivery and to assess the implications for acceptability and utilisation of maternity services in Ghana. METHODS: Twenty-one individual in-depth interviews and two focus group discussions were conducted with women of reproductive age who had delivered in the past five years in the Greater Accra Region. The study investigated women's perceptions and experiences of care in terms of factors that influenced place of delivery, satisfaction with services, expectations of care and whether they would recommend services. RESULTS: One component of care which appeared to be of great importance to women was staff attitudes. This factor had considerable influence on acceptability and utilisation of services. Otherwise, a successful labour outcome and non-medical factors such as cost, perceived quality of care and proximity of services were important. Our findings indicate that women expect humane, professional and courteous treatment from health professionals and a reasonable standard of physical environment. Women will consciously change their place of delivery and recommendations to others if they experience degrading and unacceptable behaviour. CONCLUSION: The findings suggest that inter-personal aspects of care are key to women's expectations, which in turn govern satisfaction. Service improvements which address this aspect of care are likely to have an impact on health seeking behaviour and utilisation. Our findings suggest that user-views are important and warrant further investigation. The views of providers should also be investigated to identify channels by which service improvements, taking into account women's views, could be operationalised. We also recommend that interventions to improve delivery care should not only be directed to the health professional, but also to general health system improvements. PMID- 16372912 TI - Portable light transmission measuring system for preserved corneas. AB - BACKGROUND: The authors have developed a small portable device for the objective measurement of the transparency of corneas stored in preservative medium, for use by eye banks in evaluation prior to transplantation. METHODS: The optical system consists of a white light, lenses, and pinholes that collimate the white light beams and illuminate the cornea in its preservative medium, and an optical filter (400-700 nm) that selects the range of the wavelength of interest. A sensor detects the light that passes through the cornea, and the average corneal transparency is displayed. In order to obtain only the tissue transparency, an electronic circuit was built to detect a baseline input of the preservative medium prior to the measurement of corneal transparency. The operation of the system involves three steps: adjusting the "0 %" transmittance of the instrument, determining the "100 %" transmittance of the system, and finally measuring the transparency of the preserved cornea inside the storage medium. RESULTS: Fifty selected corneas were evaluated. Each cornea was submitted to three evaluation methods: subjective classification of transparency through a slit lamp, quantification of the transmittance of light using a corneal spectrophotometer previously developed, and measurement of transparency with the portable device. CONCLUSION: By comparing the three methods and using the expertise of eye bank trained personnel, a table for quantifying corneal transparency with the new device has been developed. The correlation factor between the corneal spectrophotometer and the new device is 0,99813, leading to a system that is able to standardize transparency measurements of preserved corneas, which is currently done subjectively. PMID- 16372913 TI - Long-term air pollution exposure and living close to busy roads are associated with COPD in women. AB - BACKGROUND: Lung function and exacerbations of chronic obstructive pulmonary disease (COPD) have been associated with short-term exposure to air pollution. However, the effect of long-term exposure to particulate matter from industry and traffic on COPD as defined by lung function has not been evaluated so far. Our study was designed to investigate the influence of long-term exposure to air pollution on respiratory symptoms and pulmonary function in 55-year-old women. We especially focused on COPD as defined by GOLD criteria and additionally compared the effects of air pollution on respiratory symptoms by questionnaire data and by lung function measurements. METHODS: In consecutive cross sectional studies conducted between 1985-1994, we investigated 4757 women living in the Rhine-Ruhr Basin of Germany. NO2 and PM10 exposure was assessed by measurements done in an 8 km grid, and traffic exposure by distance from the residential address to the nearest major road using Geographic Information System data. Lung function was determined and COPD was defined by using the GOLD criteria. Chronic respiratory symptoms and possible confounders were defined by questionnaire data. Linear and logistic regressions, including random effects were used to account for confounding and clustering on city level. RESULTS: The prevalence of COPD (GOLD stages 1-4) was 4.5%. COPD and pulmonary function were strongest affected by PM10 and traffic related exposure. A 7 microg/m3 increase in five year means of PM10 (interquartile range) was associated with a 5.1% (95% CI 2.5%-7.7%) decrease in FEV1, a 3.7% (95% CI 1.8%-5.5%) decrease in FVC and an odds ratio (OR) of 1.33 (95% CI 1.03-1.72) for COPD. Women living less than 100 m from a busy road also had a significantly decreased lung function and COPD was 1.79 times more likely (95% CI 1.06-3.02) than for those living farther away. Chronic symptoms as based on questionnaire information showed effects in the same direction, but less pronounced. CONCLUSION: Chronic exposure to PM10, NO2 and living near a major road might increase the risk of developing COPD and can have a detrimental effect on lung function. PMID- 16372914 TI - Targeting the active site of the placental isozyme of alkaline phosphatase by phage-displayed scFv antibodies selected by a specific uncompetitive inhibitor. AB - BACKGROUND: The isozymes of alkaline phosphatase, the tissue non-specific, intestinal and placental, have similar properties and a high degree of identity. The placental isozyme (PLAP) is an oncofetal antigen expressed in several malignancies including choriocarcinoma, seminoma and ovarian carcinoma. We had earlier attempted to isolate PLAP-specific scFv from a synthetic human immunoglobulin library but were unable to do so, presumably because of the similarity between the isozymes. In this work, we have employed a PLAP-specific uncompetitive inhibitor, L-Phe-Gly-Gly, to select isozyme specific scFvs. An uncompetitive inhibitor binds to the enzyme in the presence of substrate and stabilizes the enzyme-substrate complex. Several uncompetitive inhibitors have varying degrees of isozyme specificity for human alkaline phosphatase isozymes. A specific uncompetitive inhibitor would be able to unmask conformational differences between the otherwise very similar molecules. Also, such inhibitors would be directed to regions at/close to the active site of the enzyme. In this work, the library was first incubated with PLAP and the bound clones then eluted by incubation with L-Phe-Gly-Gly along with the substrate, para-nitro phenyl phosphate (pNPP). The scFvs were then studied with regard to the biochemical modulation of their binding, isozyme specificity and effect on enzyme activity. RESULTS: Of 13 clones studied initially, the binding of 9 was inhibited by L-Phe Gly-Gly (with pNPP) and 2 clones were inhibited by pNPP alone. Two clones had absolute and 2 clones had partial specificity to PLAP. Two clones were cross reactive with only one other isozyme. Three scFv clones, having an accessible His6-tag, were purified and studied for their modulation of enzyme activity. All the three scFvs inhibited PLAP activity with the kinetics of competitive inhibition. Cell ELISA could demonstrate binding of the specific scFvs to the cell surface expressed PLAP. CONCLUSION: The results demonstrate the biochemical modulation of scFv binding. Also, the scFvs bound to the active site and denied the access to the substrate. The selection strategy could generate specific anti enzyme antibodies to PLAP that can potentially be used for targeting, for modulating enzyme activity in in vitro and in vivo and as probes for the active site. This strategy also has a general application in selecting antibodies from combinatorial libraries to closely related molecules and conformations. PMID- 16372916 TI - Report of a pre ICN workshop on negotiating the future of nutrition, Johannesburg, South Africa, 18 September 2005. PMID- 16372917 TI - Out of the Christmas box. PMID- 16372918 TI - Challenges in translating scientific evidence into mandatory food fortification policy: an antipodean case study of the folate-neural tube defect relationship. AB - OBJECTIVE: To identify challenges in translating scientific evidence of a nutrient and health relationship into mandatory food fortification policy. DESIGN: A case study approach was used in which available evidence associated with the folate-neural tube defect relationship was reviewed against the Australia New Zealand Food Regulation Ministerial Council's Policy Guideline for mandatory food fortification. RESULTS: Three particular challenges were identified. The first is knowing when and how to act in the face of scientific uncertainty. The second is knowing how to address the special needs of at-risk individuals without compromising the health and safety of the population as a whole. The third is to ensure that a policy is sufficiently monitored and evaluated. CONCLUSIONS: Despite the availability of compelling evidence of a relationship between a particular nutrient and a health outcome, a definitive policy response may not be apparent. Judgement and interpretation inevitably play significant roles in influencing whether and how authorities translate scientific evidence into mandatory food fortification policy. In relation to the case study, it would be prudent to undertake a risk-benefit analysis of policy alternatives and to implement nutrition education activities to promote folic acid supplement use among the target group. Should mandatory folate fortification be implemented, comprehensive monitoring and evaluation of this policy will be essential to know that it is implemented as planned and does more good than harm. In relation to mandatory food fortification policy-making around the world, ongoing national nutrition surveys are required to complement national policy guidelines. PMID- 16372919 TI - Taxing food: implications for public health nutrition. AB - AIM: To set out a policy analysis of food taxes as a way of influencing food consumption and behaviour. DESIGN: The study draws on examples of food taxes from the developed world imposed at national and local levels. Studies were identified from a systemised search in six databases with criteria designed to identity articles of policy relevance. RESULTS: The dominant approach identified from the literature was the imposition of food taxes on food to raise general revenue, such as Value Added Tax in the European Union. Food taxes can be applied in various ways, ranging from attempts to directly influence behaviour to those which collect taxes for identified campaigns on healthy eating through to those applied within closed settings such as schools. There is a case for combining taxes of unhealthy foods with subsidies of healthy foods. The evidence from the literature concerning the use and impact of food taxes on food behaviour is not clear and those cases identified are mainly retrospective descriptions of the process. Many food taxes have been withdrawn after short periods of time due to industry lobbying. CONCLUSIONS FOR POLICY: Small taxes with the clear purpose of promoting the health of key groups, e.g. children, are more likely to receive public support. The focus of many tax initiatives is unclear; although they are generally aimed at consumers, another focus could be food manufacturers, using taxes and subsidies to encourage the production of healthier foods, which could have an effect at a population level. Further consideration needs to be given to this aspect of food taxes. Taxing food (and subsidies) can influence food behaviour within closed systems such as schools and the workplace. PMID- 16372920 TI - The impact of a national mass media campaign in The Netherlands on the prevention of weight gain. AB - OBJECTIVE: A 5-year nationwide mass media campaign aimed at prevention of overweight was organised from 2002 onwards. The present study evaluates the first campaign, which was aimed primarily at increasing awareness of weight gain. DESIGN AND SUBJECTS: Data were collected by telephone interview in four independent cross-sectional surveys among non-obese Dutch adults aged 25-35 years (total n = 1,949) for statistical analyses. Awareness of personal body-weight status, overweight-related risk perceptions, attitudes towards weight-gain prevention, motivation to prevent weight gain and self-reported body mass index (BMI) were measured in each survey. Campaign exposure was assessed in the post intervention surveys. To identify intervention effects over time multiple linear and logistic regression analyses were used, adjusted for secular time effects and age. RESULTS: After the campaign about 65% of the respondents knew about the campaign. The campaign was associated with more positive attitudes towards the prevention of weight gain (beta = 0.16; P < 0.01) and higher self-reported BMI (beta = 0.14; P < 0.01). CONCLUSIONS: The results suggest that the first campaign reached a large proportion of the population and initiated some positive change in attitudes, but did not achieve significant improvements in other determinants of weight-gain prevention among non-obese young adults. PMID- 16372921 TI - Why do adolescents eat what they eat? Personal and social environmental predictors of fruit, snack and breakfast consumption among 12-14-year-old Dutch students. AB - OBJECTIVE: The aim of the study was to investigate the relative importance of personal and social environmental predictors of the consumption of fruit, high fat snacks and breakfast. DESIGN: A school-based cross-sectional survey. Data were collected through written questionnaires. SETTING: Students from eight schools in the southern part of The Netherlands. SUBJECTS: Six hundred and one students from preparatory secondary vocational education schools. RESULTS: About a quarter of the variation in actual behaviours and intentions to change the behaviours could be explained. Stepwise multiple regression analysis revealed that, for all three behaviours, higher intention to change was especially associated with a more positive attitude and subjective norm, and higher intentions to increase fruit intake with more positive self-efficacy expectations. With regard to actual consumption, a more positive attitude towards eating fruit was the only significant correlate of a higher consumption of fruit. A more positive attitude towards eating high-fat snacks, perceived lower intake of the mother, and higher food availability and accessibility were associated with consumption of high-fat snacks, and a more positive attitude to breakfast more frequently was associated with more frequent breakfast consumption. CONCLUSION: The results indicate that adolescents' attitudes are the most important determinants of different health-related eating behaviours and intentions to change. Interventions promoting a healthy diet for adolescents should include creative strategies to achieve positive associations with healthy dietary changes. PMID- 16372922 TI - Effects of retirement on lifestyle in relation to changes in weight and waist circumference in Dutch men: a prospective study. AB - OBJECTIVE: To study changes in lifestyle in relation to changes in body weight and waist circumference associated with occupational retirement in men. DESIGN: A prospective cohort study with 5 years of follow-up. At baseline and at follow-up, questionnaires were completed and body weight and waist circumference were measured. SETTING: The Doetinchem Cohort Study, consisting of inhabitants of Doetinchem, a town in a rural area of The Netherlands. SUBJECTS: In total 288 healthy men aged 50-65 years at baseline, who either remained employed or retired over follow-up. RESULTS: The effect of retirement on changes in weight and waist circumference was dependent on type of former occupation. Increase in body weight and waist circumference was higher among men who retired from active jobs (0.42 kg year(-1) and 0.77 cm year(-1), respectively) than among men who retired from sedentary jobs (0.08 kg year(-1) and 0.23 cm year(-1), respectively). Weight gain and increase in waist circumference were associated with a decrease in fruit consumption and fibre density of the diet, with an increase in frequency of eating breakfast, and with a decrease in several physical activities, such as household activities, bicycling, walking and doing odd jobs. CONCLUSION: Retirement was associated with an increase in weight and waist circumference among those with former active jobs, but not among those with former sedentary jobs. Retirement may bring opportunities for healthy changes in diet and physical activity, which could be used in health promotion programmes. PMID- 16372923 TI - Is the prevalence of dehydration among community-dwelling older adults really low? Informing current debate over the fluid recommendation for adults aged 70+years. AB - OBJECTIVE: The fluid recommendation for adults aged 70+ years has been criticised on the basis of a low prevalence of dehydration in community-dwelling older adults. This study explores whether the low prevalence might reflect limitations of individual dehydration indices. DESIGN: Cross-sectional data on plasma sodium, blood urea nitrogen (BUN), creatinine, glucose and potassium were used to classify 1,737 participants of the 1992 Established Populations for Epidemiologic Studies of the Elderly (EPESE) (70+ years) according to multiple dehydration indices. Associations between dehydration indices, health and functional status were evaluated. RESULTS: Depending on the indicator used, the prevalence of dehydration ranged from 0.5% for hypotonic hypovolaemia only (plasma tonicity < 285 mOsm l(-1) with orthostatic hypotension) to 60% with dehydration defined as either plasma sodium >or=145 mEq l(-1), BUN/creatinine ratio >or=20, tonicity >or=295 mOsm l(-1), or hypotonic hypovolaemia. Elevated tonicity and BUN/creatinine ratio were respectively associated with chronic disease and functional impairment. CONCLUSIONS: The true prevalence of dehydration among community-dwelling adults may be low or high, depending on the indicator(s) used to define dehydration. Before we can pinpoint a generalisable prevalence of dehydration for community-dwelling seniors and draw conclusions about fluid recommendations, validation studies of dehydration indices and longitudinal studies of dehydration, health and functional status are needed. PMID- 16372924 TI - Factors determining the nutrition transition in two Mediterranean islands: Sardinia and Malta. AB - OBJECTIVE: To examine and analyse consumption changes over time of 24 food items between Sardinia and Malta. SETTING: The data were collected in 2001 in Sardinia and 2002 in Malta. DESIGN: A structured qualitative questionnaire, articulated around four main themes: food supply, transformation, preparation and consumption habits, was administered by face-to-face interviews with the help of a local person. It encompassed mainly open-ended questions, which allowed us to measure factors contributing to change. SUBJECTS: Thirty mother-daughter pairs were interviewed in each insular territory. RESULTS: Despite a common trend revealing a shift away from cereals, pulses and potatoes to the benefit of meat products, fats and sugar, our results showed contrasting evolutions in food consumption between both insular societies. Fruit and vegetables, olive oil and fish, which are part of the main features of the Mediterranean diet, were among the top foods for which consumption frequency has increased in Sardinia. In Malta, besides an increase in olive oil and vegetable consumption, cheeses and desserts showed the highest increase. Along with modernity and improved living conditions, enhanced commercial availability and increased diversity of food preparation were also identified as factors contributing to food consumption changes. CONCLUSIONS: Although the Sardo-Mediterranean model is evolving under the impact of modernisation, it is not disappearing. In Malta, however, modernity has led to a more sudden shift from a state of food shortage to one of affluence, but in a cultural context where the identity is no longer Mediterranean but Anglo-Saxon. PMID- 16372925 TI - Sources of variation in nutrient intakes among men in Shanghai, China. AB - BACKGROUND AND OBJECTIVE: Random errors, from any source, will attenuate epidemiological risk estimates. Before we launched the Shanghai Men's Health Study (SMHS), a large population-based cohort study investigating the diet-cancer association among Chinese men, a dietary calibration study was conducted among 96 men aged 40-75 years (mean age 56.5 years), with biweekly 24-hour dietary recalls (24HDRs) implemented over a 1-year period. Data from this study were analysed to evaluate the nature and magnitude of variances for intake of 26 nutrients among SMHS participants, to compare variance ratios of 26 nutrients among Chinese men and women and individuals in other studies, and to estimate the number of 24HDRs required for future dietary calibration studies in similar populations. DESIGN: Ninety-six healthy, free-living men in Shanghai were administered biweekly 24HDR interviews 24 times over a 1-year period. To assess between-individual and within individual contributions to variance, a mixed effects model was fitted and ratios of within-individual to between-individual (sigma(w)(2)/sigma(b)(2)) dietary intake variances were computed. SETTING: Shanghai, China. RESULTS: In agreement with reports from studies conducted in the USA and many other countries, we found that within-individual variances were usually larger than between-individual variances in dietary intake for all nutrients. The sum of all other variation (e.g. weekday and weekend, seasonal, interviewer) accounted for less than 5% of total variation. Ratios of within- to between-individual variances (for log transformed data) ranged from 1.25 for carbohydrate intake to near 8 for delta tocopherol intake. CONCLUSIONS: The results of this study suggest that among middle-aged and elderly Chinese men in Shanghai, within- and between-individual variation account for more than 95% of the total variation for 26 nutrients. Further dietary validation studies in the same population could be adequately carried out with only 12 days of dietary recalls, if 100 participants were enrolled. PMID- 16372926 TI - Green tea consumption in everyday life and mental health. AB - OBJECTIVE: Green tea has been widely acknowledged in Japan to induce a pleasurable mental feeling. Recent laboratory studies have suggested positive psychological effects as a result of consuming green tea. The present study examined whether green tea consumption in everyday life in Japan is associated with positive mental health. DESIGN: A cross-sectional study was performed in February-March 2002. SETTING AND SUBJECTS: The subjects of the study consisted of a general population of 600 Japanese aged 20-69 years. Responses of 380 subjects, obtained by home-visit interview, were analysed. The questionnaire inquired about consumption of brewed green tea and other beverages, perceived mental health status, lifestyle and others. The 12-item General Health Questionnaire (GHQ 12) was used for the assessment of mental ill-health (GHQ score >or=4). RESULTS: After adjustments for age, area, perceived mental stress, lifestyle and daily caffeine intake, the consumption of brewed green tea was not statistically associated with any decrease in risk of mental ill-health among either males or females (odds ratio (OR)=0.78, 95% confidence interval (CI)=0.47-1.29 for males; OR=0.77, 95% CI=0.51-1.14 for females). Daily caffeine intake (100 mg) inclusive of green tea, black tea, coffee and other caffeine-containing beverages was associated with a higher risk of mental ill-health among females (OR=1.26, 95% CI=1.01-1.56). CONCLUSIONS: The results provide population-based evidence on the consumption of brewed green tea in everyday life and mental health, together with information on consumption patterns of various beverages and lifestyles. PMID- 16372927 TI - A population-based case-control study of diet and melanoma risk in northern Italy. AB - OBJECTIVE: We aimed at examining the association between dietary constituents and risk of cutaneous melanoma. DESIGN: In an area of northern Italy we recruited 59 newly diagnosed melanoma patients and 59 age- and sex-matched population controls, to whom we administered a validated semi-quantitative food-frequency questionnaire. RESULTS: We found an excess risk of melanoma in subjects with a higher energy-adjusted intake of total polyunsaturated fatty acids and, in particular, of linoleic acid (relative risk = 2.16 for intake in the highest tertile compared with the lowest tertile, P for linear trend = 0.061). Conversely, disease risk was inversely associated with the consumption of soluble carbohydrates (relative risk = 0.34 for intake in the upper vs. the lowest tertile adjusting for total energy intake, P for linear trend = 0.046). No other dietary factors, including alcohol, vitamins and trace elements, correlated with melanoma risk. The association of melanoma risk with linoleic acid and soluble carbohydrates intakes was further strengthened in multivariate analysis, and when analysis was limited to females. CONCLUSIONS: Overall, these results indicate that an excess energy-adjusted intake of linoleic acid and a lower consumption of soluble carbohydrates may increase melanoma risk. PMID- 16372928 TI - Validation of a pre-coded food diary used among 13-year-olds: comparison of energy intake with energy expenditure. AB - OBJECTIVE: To validate energy intake (EI) estimated from pre-coded food diaries against energy expenditure (EE) measured with a validated position-and-movement monitor (ActiReg) in groups of 13-year-old Norwegian schoolchildren. DESIGN: Two studies were conducted. In study 1 the monitoring period was 4 days; participants recorded their food intake for four consecutive weekdays using food diaries and wore the ActiReg during the same period. In study 2 the monitoring period was 7 days; participants recorded their food intake for four consecutive days but wore the ActiReg for a whole week. SETTINGS: Participants were recruited from grade 8 in a school in and one outside Oslo (Norway). SUBJECTS: Forty-one and 31 participants from study 1 and 2, respectively, completed the study. RESULTS: The group average EI was 34% lower than the measured EE in study 1 and 24% lower in study 2. The width of the 95% confidence limits of agreement in a Bland-Altman plot for EI and EE varied from -0.2 MJ to 8.2 MJ in study 1 and from -2.3 MJ to 6.9 MJ in study 2. The Pearson correlation coefficients between reported energy intake and expenditure were 0.47 (P=0.002) in study 1 and 0.74 (P<0.001) in study 2. CONCLUSION: The data showed that there was substantial variability in the accuracy of the food diary at the individual level. Furthermore, the diary underestimated the average energy intake. The ability of the food diary to rank individuals according to energy intake was found to be good in one of the studies and moderate in the other. PMID- 16372929 TI - Can adverse effects of dietary fat intake be overestimated as a consequence of dietary fat underreporting? AB - OBJECTIVE: To describe the consequences of systematic reporting bias by the obese for diet-disease relationships. DESIGN: The present report used 24-hour urinary nitrogen and estimates of 24-hour energy expenditure to assess error in diet reporting, and examined the consequence of accounting for this error for associations between dietary fat intake and serum low-density lipoprotein (LDL) cholesterol. SETTING: Sub-study to the Danish MONICA (Monitoring Trends and Determinants in Cardiovascular Disease) project, carried out in 1987-1988. SUBJECTS: A random sub-sample of the adult Danish male population (n = 152). RESULTS: Correcting dietary fat for underreporting error weakened, rather than strengthened, the association between dietary fat intake and LDL-cholesterol by reducing the slope of the regression from beta = 3.4, P = 0.02 to beta = 2.7, P = 0.04. CONCLUSION: This example illustrates that systematic underreporting of dietary fat by high-risk groups such as the obese may produce an overestimated association. These results imply that previous epidemiological studies showing a positive association between percentage of energy from fat and other health outcomes, e.g. cancer and heart disease, may have overestimated the negative effects of a high-fat diet. If we were able to correctly assess dietary fat intake in general populations, recommendations for fat intake may be more liberal than the 30% suggested today. Improved assessment of fat intake in epidemiological studies is necessary for future development of evidence-based recommendations for diet and health . PMID- 16372930 TI - Obesity and related medical conditions: a role for functional foods. PMID- 16372931 TI - Why people see things that are not there: a novel Perception and Attention Deficit model for recurrent complex visual hallucinations. AB - As many as two million people in the United Kingdom repeatedly see people, animals, and objects that have no objective reality. Hallucinations on the border of sleep, dementing illnesses, delirium, eye disease, and schizophrenia account for 90% of these. The remainder have rarer disorders. We review existing models of recurrent complex visual hallucinations (RCVH) in the awake person, including cortical irritation, cortical hyperexcitability and cortical release, top-down activation, misperception, dream intrusion, and interactive models. We provide evidence that these can neither fully account for the phenomenology of RCVH, nor for variations in the frequency of RCVH in different disorders. We propose a novel Perception and Attention Deficit (PAD) model for RCVH. A combination of impaired attentional binding and poor sensory activation of a correct proto object, in conjunction with a relatively intact scene representation, bias perception to allow the intrusion of a hallucinatory proto-object into a scene perception. Incorporation of this image into a context-specific hallucinatory scene representation accounts for repetitive hallucinations. We suggest that these impairments are underpinned by disturbances in a lateral frontal cortex ventral visual stream system. We show how the frequency of RCVH in different diseases is related to the coexistence of attentional and visual perceptual impairments; how attentional and perceptual processes can account for their phenomenology; and that diseases and other states with high rates of RCVH have cholinergic dysfunction in both frontal cortex and the ventral visual stream. Several tests of the model are indicated, together with a number of treatment options that it generates. PMID- 16372952 TI - "Economic man" in cross-cultural perspective: behavioral experiments in 15 small scale societies. AB - Researchers from across the social sciences have found consistent deviations from the predictions of the canonical model of self-interest in hundreds of experiments from around the world. This research, however, cannot determine whether the uniformity results from universal patterns of human behavior or from the limited cultural variation available among the university students used in virtually all prior experimental work. To address this, we undertook a cross cultural study of behavior in ultimatum, public goods, and dictator games in a range of small-scale societies exhibiting a wide variety of economic and cultural conditions. We found, first, that the canonical model - based on self-interest - fails in all of the societies studied. Second, our data reveal substantially more behavioral variability across social groups than has been found in previous research. Third, group-level differences in economic organization and the structure of social interactions explain a substantial portion of the behavioral variation across societies: the higher the degree of market integration and the higher the payoffs to cooperation in everyday life, the greater the level of prosociality expressed in experimental games. Fourth, the available individual level economic and demographic variables do not consistently explain game behavior, either within or across groups. Fifth, in many cases experimental play appears to reflect the common interactional patterns of everyday life. PMID- 16372980 TI - Audit of specialist registrar training in tympanomastoid surgery for chronic otitis media. AB - BACKGROUND: A prospective audit of specialist registrars' (SRs') training in tympanomastoid surgery for chronic otitis media within the Anglia Regional Training Scheme is described. This audit recorded the surgical activity of the trainees and their contribution to operative procedures, and assessed the results of the procedures. This type of systematic approach to the audit of surgical training is important in light of the current shortened training programmes and increased accountability of trainers. OBJECTIVES: The study aimed to establish the levels of exposure to, supervision of and outcome of ear operations for chronic otitis media performed by ENT trainees in the East Anglia region. METHOD: A prospective, region-wide, minimum otology dataset-based proforma audit was undertaken, with compulsory SR participation. Proformas were completed at the time of operation (form one) and at a minimum interval of nine months post operatively (form two). Data on form one included hospital, supervising consultant, name and training year of SR, contribution of SR (based on England Royal College of Surgeons guidelines interpreted by the SR), pre-operative audiology average (air conduction/bone conduction over 0.5, 1, 2 and 4 kHz), the pathology and the state of the ear at the time of surgery, and a breakdown of the procedure(s) undertaken. Form two recorded data relevant to form one as well as information regarding patient satisfaction and the operative result obtained, graded as 'gold' (no disease, dry ear and hearing average < 25 dB), 'silver' (two of these three) and 'bronze' (one of these three). All completed forms were analysed using Microsoft Access software. RESULTS: Completed copies of 409 form ones and 156 form twos were analysed. With advancing years, SRs' contributions to procedures increased without significant effect on the graded outcome, which appeared to be independent of SR year of training. Different regional hospitals were compared. Data collected also provided an otology training portfolio for SRs, forming part of their registrar in-training assessment (RITA). CONCLUSION: The East Anglia SR audit of SRs' training in tympanomastoid surgery for chronic otitis media was a powerful training tool. It demonstrated the safe progression of SR training in supervised ear surgery, with SRs' results being comparable to those for consultant-performed procedures. PMID- 16372982 TI - Reviewing the reviews: some thoughts from the JLO statistical advisor. PMID- 16372985 TI - Extracranial internal carotid artery aneurysm presenting as pharyngeal mass with dysphagia. AB - Aneurysms of the extracranial portion of the internal carotid artery are rare. Generally, they occur just at the level of, or above, the bifurcation. Here we report a case of a left internal carotid artery aneurysm presenting as an oropharyngeal mass causing dysphagia. PMID- 16372986 TI - Coexistence of laryngeal squamous cell carcinoma and non-Hodgkin's lymphoma with nasopharyngeal involvement. AB - Laryngeal squamous cell carcinoma (SCC) is one of the most frequent malignancies in the head and neck region. The risk of multiple malignancies is reported as 2 11 per cent and most of the second primaries are SCCs. Lymphogenic tumours as second primaries are extremely rare. In this paper, we report a case of laryngeal SCC with synchronous non-Hodgkin's lymphoma and review the literature on the clinical and histopathological aspects of these malignancies. PMID- 16372987 TI - Reconstruction of the tensor tympani tendon. AB - We describe a case in which reconstruction of the tendon of the tensor tympani muscle was necessary for the successful restoration of sound conduction. The right ear of a nine-year-old boy was treated for cholesteatoma with staged surgery. During the first operation, the tendon was cut to ensure good visibility in the tympanic cavity. Post-operatively, maintenance of aeration of the middle ear required ventilation tubes at first and Valsalva manoeuvres later on. The position of the reconstructed tympanic membrane varied a great deal, moving between the medial wall of the tympanic cavity and extreme bulging. This made exact measurement of a columella for ossicular reconstruction impossible. The preserved handle of the malleus was bound to the cochleariform process with ionomer cement, using a piece of surgical suture material as a substitute for the tendon. This arrangement prevented the tympanic membrane from undergoing excessive lateral movement after inflation and the ossicular chain was replaced with a successful ossiculoplasty with an autogenous bone 'drum to footplate' columella. The pre-operative 55.0 dB air-bone gap decreased immediately to 3.3 dB, widening after three years to 15.0 dB. PMID- 16372988 TI - Interdigitating dendritic cell sarcoma of the parotid gland. AB - Interdigitating dendritic cell sarcomas (IDCSs) are extremely uncommon tumours that arise predominantly in lymphoid tissue. We report a case of an IDCS arising in the parotid gland of a 73-year-old man. Clinically, a primary salivary gland tumour was suspected but fine needle aspiration cytology suggested a soft tissue tumour. A diagnosis of IDCS was made on histopathological examination of the resection specimen, with subsequent confirmation by electron microscopy. Given the extreme rarity of this tumour at this site, it is unlikely to be a common diagnostic problem, but the importance of multiple diagnostic modalities is emphasized. The findings of cytology, histology, immunohistochemistry and electron microscopy have not previously been described together in a single case report of this tumour. PMID- 16372989 TI - Inflammatory nasal polyps: an unusual late complication of Silastic sheet repair of orbital floor fracture. AB - Silastic implants are very widely used in surgical practice and are considered to be relatively inert. They do however present with complications, including infection, local foreign body inflammatory response,calcification, migration and failure of repair of the defect, which sometimes may necessitate explantation. Head and neck implants do present a special case, as complications can cause obstruction and disruption of function in small cavities. A pertinent history, clinical review and computed tomography scan are usually invaluable in obtaining a diagnosis. We present a rare case of migrated Silastic orbital sheet, presenting as a nasal polyp and causing maxillary antral pain and infection. A detailed search of the medical literature revealed no other such case. PMID- 16372990 TI - Effect of nasal septal deviation on total ethmoid cell volume. AB - BACKGROUND: The aim of this study was to evaluate the effect of nasal septal deviation (NSD) on ethmoid cell volume and to determine whether there was any correlation between NSD grade and ethmoid cell volume. METHODS: Forty computerized tomography (CT) scans from patients with rhinosinusitis symptoms with NSD were evaluated. Septal deviations were classified into three groups according to the degree of deviation on CT. Ethmoid cell volumes were measured and the relationship between NSD and ethmoid cell volume was investigated. RESULTS: There was a moderate but significant negative correlation between the septal deviation angle and the percentage of the ethmoid cell volumes (p = 0.001, r = -0.5152, r2 = 0.2654). Total ethmoid cell volume on the ipsilateral side compared with the contralateral side was found to decrease as the degree of NSD increased. CONCLUSIONS: Nasal septal deviation affects the total ethmoid cell volume of the nasal cavity. The results of our study underline the role of ethmoid cell volume in the compensation mechanism equalizing the nasal cavity airflow changes due to NSD. PMID- 16372991 TI - Laryngeal dysplasia: aetiology and molecular biology. AB - Laryngeal premalignancy is a common clinical concern. While tobacco has long been established as the principal identifiable aetiological factor, the last two decades have seen publications investigating a potential role for gastroesophageal reflux, gastrectomy and human papilloma virus. Furthermore, there have been major advances in our understanding of the molecular biology of cancer and premalignancy. Accompanying this increased understanding, significant efforts have been made to correlate the expression of molecular markers with the clinical course of premalignant laryngeal lesions. This review summarizes current knowledge of the aetiological factors and molecular biology of laryngeal premalignancy. All aetiological factors, including molecular markers, are discussed separately and their possible role in the clinical course of these lesions is discussed. PMID- 16372992 TI - Acute haemorrhage in patients with advanced head and neck cancer: value of endovascular therapy as palliative treatment option. AB - AIMS: Acute or subacute haemorrhage is one of the most frightening complications in patients suffering from advanced head and neck cancer. Few articles report experience with superselective endovascular therapy for this purpose. Is endovascular therapy underestimated in the field of palliative head and neck cancer therapy? This study set out to investigate this question. PATIENTS AND METHODS: A review was undertaken of the clinical courses of seven patients (six men, one woman) suffering from incurable, advanced head and neck cancer (four pharyngeal, two laryngeal, one neck) and treated with superselective endovascular strategies as an emergency procedure for acute bleeding. RESULTS: All patients were successfully treated without evidence of neurological complication. Patients reached a median survival of 20 weeks (range eight-168 weeks). Following endovascular treatment all patients were discharged from the hospital within several days. Three patients survived almost free of symptoms for several weeks and were able to stay at home with their families until their death. CONCLUSION: We conclude that in the field of palliative care, superselective endovascular therapy deserves to be considered alongside standard treatment options for the management of acute haemorrhage from advanced head and neck cancer. PMID- 16372993 TI - Leiomyosarcoma of the larynx. AB - Smooth muscle tumours of the head and neck are rare. An unusual case of laryngeal leiomyosarcoma in a 69-year-old man is presented. Histological diagnosis of this tumour is difficult, and immunohistochemical studies are vital. Treatment is discussed, and should include local control with debulking, radiotherapy and long term follow up. PMID- 16372998 TI - Prevention and treatment of HIV infection in the future. PMID- 16373000 TI - Current situation in the development of a preventive HIV vaccine. AB - The uncontrolled progression of the AIDS epidemic has made the development of an efficacious human immunodeficiency virus (HIV) vaccine a major objective of scientific research. No effective preventive vaccine against HIV is currently available and sterilizing immunity has not yet been achieved in animal models. This review analyses the major challenges in developing an AIDS vaccine, in particular the mechanisms involved in viral escape from the immune response, and summarizes the results obtained with the different prototypes of therapeutic and preventive vaccines. Finally, social, economic and healthcare aspects of research into HIV vaccines and current controversies regarding the development of clinical trials are discussed. PMID- 16373002 TI - New targets and new drugs in the treatment of HIV. AB - Antiretroviral treatment has modified the course of human immunodeficiency virus (HIV) infection transforming it into a chronic disease. However, as treatment is conceived "for life", more effective and safety drugs, overcoming the growing resistance of the virus are required. New molecules may block the known viral targets or other new ones. The mechanism of the virus union and entrance to the cell includes the new therapeutic targets that are studied more frequently. Although studies with substances that efficiently block the virus-CD4 receptors union are in very early phases, other studies of molecules capable to block the entrance co-receptors are in more advanced phases (II or III), and enfuvirtide, a substance that blocks membrane fusion, the last phase of virus entrance, has been recently marketed. Another very promising pharmacological target is the integration of the proviral DNA as we know some substances that in vitro block HIV integrase. Besides this, new drugs are increasing the three classic antiretroviral families. Among nucleoside analogs emtricitabine (recently marketed) and amdoxovir are the more prominent. Capravirine and TMC-125 are the non-nucleoside analogs whose studies are more advanced. And atazanavir, fos amprenavir, tipranavir and TMC-114 are the new protease inhibitors recently marketed or near to be. PMID- 16373004 TI - Therapeutic drug monitoring and the inhibitory quotient of antiretroviral drugs: can they be applied to the current situation? AB - Therapeutic drug monitoring is attracting growing interest as a means of increasing the effectiveness of antiretroviral therapy and of decreasing its toxicity, although data supporting this strategy are still scarce. Currently, nucleoside analog reverse-transcriptase inhibitors (NARTI) are not candidates because their effect depends on their active intracellular form and not on plasma concentration. Protease inhibitors (PI) and non-nucleoside analogue reverse transcriptase inhibitors (NNARTI) meet the criteria for therapeutic drug monitoring. The main limitations are that the parameters to be monitored in order to measure exposure to the drug and the effective concentration of the drug have not been well defined. The few studies performed in treatment-naive patients have demonstrated that monitoring improves therapeutic efficacy. This strategy will be particularly useful when the risk of subtherapeutic or toxic concentrations is especially high (pharmacokinetic interactions, intestinal malabsorption, adverse effects, virological failure without obvious cause, pregnancy, children). Although it remains to be standardized, the inhibitory quotient integrates pharmacological and virological parameters and is useful in patients with prior virological failure. Any therapeutic drug monitoring program should be accompanied by measures to monitor and improve treatment adherence. There are good reasons to believe that therapeutic drug monitoring can be useful to improve treatment in specific circumstances. However, before its widespread use as a routine method can be recommended, the parameters to be used should be standardized and studies with appropriate methodology should be performed to define the role of therapeutic drug monitoring in distinct clinical situations. PMID- 16373006 TI - The future of HIV infection: gene therapy and RNA interference. AB - The description of the mechanism of RNA interference (RNAi) has generated enormous interest in the biomedical field. A previously unrecognized pathway in which small interfering, 21 to 23 mer, double-stranded RNA (siRNA) mediates sequence-specific degradation of mRNA is becoming one the most useful techniques in cell biology and genetics research. Based on the potency, specificity and physiology of RNAi to silence gene expression, much is expected from its use as a therapeutic tool. The first evidence of RNAi as a suppressor of HIV replication has already been reported, thus providing a new impetus to the development of molecular or gene therapy approaches to HIV infection. PMID- 16373008 TI - Immunotherapy and therapeutic vaccines in HIV infection. AB - Resistance to medication, adverse effects in the medium-long term, and cost are important limitations to lifelong adherence to highly active antiretroviral therapy (HAART). The combination of HAART with immune therapy to restore and/or boost immune-specific responses to HIV has been proposed, with the ultimate aim of controlling viral replication in the absence of HAART over long periods. The functional defects of the cellular and humoral responses would explain the lack of control of the immune system over viral replication. Different types of immune mediated therapy have been investigated to solve these problems, including passive immune therapy, cytokines, structured treatment interruptions, immunosuppressors and therapeutic vaccines. Our still limited knowledge of immune mechanisms which can control HIV viral replication and of the causes of the deterioration of cellular and humoral immunity have produced only modest benefits in immune-mediated therapy, and are therefore confined to research for the time being. The availability of an optimal therapeutic vaccine would be an important scientific advance which could be compared with the arrival of protease inhibitors in clinical practice. Therefore, priority should be given to research in this field. PMID- 16373009 TI - Cyclosporine in the treatment of severe attack of ulcerative colitis: a systematic review. AB - INTRODUCTION: Intravenous steroid therapy is the standard treatment in severe attacks of ulcerative colitis (UC), but 20% to 60% of patients fail to respond and require colectomy. Cyclosporine (CyA) has shown efficacy in steroid failures and could avoid surgery, but controversy remains. AIM: The objective of this study was to conduct a systematic review to evaluate the effectiveness and safety of CyA in inducing remission in patients with a severe attack of UC. METHODS: We did a systematic review using Cochrane methodology, including data from published (in English, French, Spanish or German) clinical trials done in adults using intravenous or oral CyA in UC. Data on efficacy are obtained from controlled and observational clinical trials, and for safety issues case reports are also considered. RESULTS: 31 studies were identified which met the inclusion criteria, 22 (18 uncontrolled, 4 controlled) with intravenous CyA, and 9 (all uncontrolled) using oral CyA. Only 4 controlled trials (one in abstract form) are available, and only one compares CyA to placebo. However, efficacy results are very consistent in these 4 trials, and very similar to those in observational studies. CyA achieves remission in 91,4% and 71.4% of patients in controlled and uncontrolled studies using intravenous route, and in 71,2% using oral route. Two mg/kg/day seems so efficacious and safer as previous standard 4 mg/kg/day dose. Minor side effects are rather common but do not seriously limit therapy. Severe side effects, specially infections, are uncommon but clinically relevant with several deaths reported. CONCLUSION: CyA (intravenous, 2 mg/kg/day) constitutes an efficacious and relatively safe alternative in the treatment of severe, steroid-refractory, attack of UC. To optimize treatment, the correct selection of patients, a standardized protocol and clinical surveillance are recommended. PMID- 16373010 TI - Evaluation of 3 different tests for the detection of stool antigens to confirm Helicobacter pylori eradication after treatment. A pilot study. AB - INTRODUCTION: Recently, several new diagnostic methods aimed to detect Helicobacter pylori stool antigens have been developed. Our aim was to evaluate the accuracy of 3 different stool tests to confirm H. pylori eradication. PATIENTS AND METHODS: Twenty-six patients received H. pylori eradication treatment. Eradication was confirmed with 13C-urea breath test 6-8 weeks later, when stool samples were analyzed by polyclonal (Premier-Platinum-HpSATM), monoclonal (Amplified-IDEIATM-HpStARTM), and rapid test (ImmunoCard-STAT-HpSATM). RESULTS: H. pylori was eradicated in 85% of the cases. Sensitivity, specificity, positive predictive value and negative predictive value with the polyclonal test were: 25%, 91%, 33% and 87%. Corresponding results with the monoclonal test, using the cut-off point recommended by the manufacturer, were 100%, 46%, 25% and 100%. However, the best cut-off point in our study had 100% sensitivity and 91% specificity. The area under ROC curve for the polyclonal and the monoclonal tests was 0.65 and 0.95. Diagnostic accuracy with the rapid test was 75%, 90%, 60% and 95%. CONCLUSION: Neither the polyclonal stool antigen test nor the rapid stool antigen test can be recommended to confirm H. pylori eradication after treatment. The monoclonal test has better diagnostic accuracy, although more studies are necessary to definitively recommend its use for the confirmation of H. pylori eradication success. PMID- 16373011 TI - Rapidly-progressive liver failure secondary to melanoma infiltration. AB - We describe the case of a 51-year-old man with a history of intraocular melanoma treated with radiotherapy 2 years previously. The patient was diagnosed with mild hypertransaminasemia that progressed to acute liver failure and death in a period of one month. Radiological investigations such as spiral computed tomography and abdominal ultrasonography failed to give an etiologic diagnosis. Autopsy revealed melanoma with diffuse infiltration of the hepatic parenchyma. Because diagnosis is usually delayed, the prognosis of intraocular melanoma is poor. In 40% of cases metastases are present at diagnosis, and the most frequently affected organ is the liver (93-95%). Presentation as acute liver failure can appear after a long disease-free period. For this reason, periodic laboratory tests and hepatic ultrasound examination are recommended in patients diagnosed with this malignancy. PMID- 16373012 TI - [Type III glycogen storage disease associated with hepatocellular carcinoma]. AB - Type III glycogen storage disease is a hereditary disorder with autosomal recessive transmission. It is characterized by accumulation of abnormal glycogen in the liver and, in 80% of patients, in muscle. The liver can also show fibrosis and sometimes cirrhosis. Until 2000, 9 cases of cirrhosis had been published, 3 of which showed associated hepatocarcinoma. We present the case of a 31-year-old woman, diagnosed in childhood with type III glycogen storage disease, who 30 years after onset developed a hepatocellular carcinoma with portal thrombosis in the context of advanced cirrhosis. This is the first case to be reported in the Spanish literature of type III glycogen storage disease associated with hepatocellular carcinoma. PMID- 16373013 TI - [Atypical presentation of distant metastases from hepatocarcinoma]. AB - Hepatocarcinoma (HCC) is the most frequent primary malignant hepatic tumour. These tumours usually develop in cirrhotic liver; for this reason, periodic screening using alphafetoprotein determination and abdominal ultrasonography is considered in cirrhotic patients with preserved hepatocellular function. This strategy allows early detection of HCC, increasing the proportion of curable tumours. The most frequent metastasic dissemination is portal vein neoplasic thrombosis, being unusual the occurrence of spread metastases in other organs. We present 3 cases of atypical HCC metastasis with non specific clinical manifestations which initial diagnosis was wrong. Because of a longer survival of these patients in recent years, spread metastasis might be considered in patients with known HCC and non specific symptoms. PMID- 16373014 TI - [Cyclosporine in ulcerative colitis flares]. PMID- 16373015 TI - [Liver disease in erythropoietic protoporphyria]. PMID- 16373016 TI - [Crigler-Najjar syndrome]. PMID- 16373017 TI - [Emerging endoscopic techniques. The arrival of virtual histology]. AB - Endoscopic technology has evolved in such a way that gastroenterologists can now visualize and store high-resolution images of the gastrointestinal tract. This has improved the approach to precancerous and cancerous lesions of the gastrointestinal tract and biliary tree. However, certain difficulties remain, especially in relation to diagnosis. In the last few years, multiple techniques have been developed that, using the properties of light, enable an instantaneous histologic diagnosis to be made while endoscopy is being performed. What has been called the "optical biopsy" allows highly exact information to be obtained, both from the morphological and functional point of view. Some of these techniques, such as chromoendoscopy and magnification, are already being performed in clinical practice while others are still under investigation. The aim of the present article is to review the underlying principles and applications of these emerging techniques. PMID- 16373018 TI - [Diastolic dysfunction in liver cirrhosis]. AB - The presence of a hyperdynamic circulation in cirrhotic liver is currently a well established concept. The first studies of cardiac function in patients with cirrhosis suggested the existence of an alcoholic cardiomyopathy. More recently, altered left ventricular response to physiological and/or pharmacological stimuli in patients with post-viral liver cirrhosis has been established, and clinically insignificant diastolic cardiac function has also been observed. Neurohumoral hyperactivity and hyperdynamic circulation, which are associated with chronic exposure to the cardiodepressant substances present in advanced liver disease, play a decisive role in the genesis of this process. The lack of relaxation of the left ventricle and alteration in the pattern of transmitral flow, both of which are characteristics of this clinical entity, are easily detected by echocardiography. The growing evidence of diastolic dysfunction in liver cirrhosis, particularly in decompensated cirrhosis, suggests the clinical importance of the problem, thus introducing the concept of "cirrhotic cardiomyopathy". Greater insight into this phenomenon could help to decrease cardiovascular risk, especially during maneuvers commonly used in the treatment of the complications of liver cirrhosis, such as paracentesis, transjugular intrahepatic portosystemic shunt stent implantation, and liver transplantation. PMID- 16373019 TI - [Hepatitis due to Chinese herbs]. PMID- 16373020 TI - [Autoimmune hepatitis in a patient with uveitis]. PMID- 16373021 TI - [Pancreatic panniculitis in a patient with metastases from pancreatic carcinoma]. PMID- 16373022 TI - [Hemodynamic response to a single dose of intravenous propranolol in the prophylaxis of variceal bleeding]. PMID- 16373023 TI - [Gastric pseudolipomatosis: an under-diagnosed entity]. PMID- 16373024 TI - [Serum C-reactive protein as a marker of outcome and infection in critical care patients]. AB - BACKGROUND AND OBJECTIVE: C-reactive protein (CRP) has been considered a marker for infection and an aid for diagnosing sepsis. We analyze the relation of CRP to infection and outcome in intensive care units (ICU) patients. PATIENTS AND METHOD: Prospective study on 77 ventilated patients. Expected short ICU stay or (suspected or confirmed) infection at admission were excluding criteria. 55 admissions after elective surgery were the controls. CRP measurement the first (CRP-1), third (CRP-3) and sixth (CRP-6) day of stay. APACHE II (Acute Physiology Score and Chronic Health Evaluation), SOFA (Sepsis-related Organ Failure Assessment), shock, respiratory or renal failure, leucocytes, platelets and albumin were registered. Follow-up until day 9 for infection and ICU discharge for outcome. RESULTS: CRP-1 in controls was 5.3 (3.9) mg/l and cases 67.8 (77.4) (p < 0.001). Shock on admission was related to CRP-1: patients in shock had higher CRP-1 levels (118.6 [82.8] vs 62.8 [75.6]; p = 0.06). 40.25% of cases developed infection, and CRP-1 levels were higher in this patients (88.8 [93.9] vs 53.8 [60.9]; p < 0.05). ROC area under curve was 0.6 with a sensibility of 23% and a specificity of 89% for a level of CRP-1 > 100. Mortality was 23.4% in cases and 1.8% in controls. Age, shock, APACHE II and SOFA were related to mortality, but CRP-1 did not. ROC area under curve for CRP-1 as mortality predictor in all patients was 0.62 (0.76 for APACHE II and 0.77 for SOFA) but only in cases was of 0.49 (0.69 for APACHE II and 0.67 for SOFA). CONCLUSIONS: CRP level on admission is an useful marker for early infection but not for outcome in critically ill patients admited to the ICU. PMID- 16373025 TI - [C-reactive protein in the acute phase of ischemic stroke]. AB - BACKGROUND AND OBJECTIVE: We aimed to assess the prognostic importance of C reactive protein (CRP) in the acute phase of ischemic stroke in-patients. PATIENTS AND METHOD: One hundred and seventeen patients within 48 h after index ischemic stroke were included. CRP levels and blood samples were obtained at this time, and a brain computerized tomography or magnetic resonance imaging were performed. Neurological and functional disability were evaluated and patients were divided according to the outcome into the following categories: transient ischemic attack, favorable stroke, and non-favorable stroke. RESULTS: 32 in patients were classified as transient ischemic attack, 31 as favourable stroke, and 54 as non-favorable stroke. There was a worsening in neurological (p < 0.0001) and functional (p < 0.005) disabilities from the TIA group to non favorable stroke. The CRP mean, by category, was 1.7, 1.07 and 3.6 mg/dl, respectively (p < 0.0001). CONCLUSIONS: We found increased levels of CRP in the non-favorable stroke category, that was related with neurological and functional disabilities, and with radiological findings, mainly when levels were greater than 3.6 (0.49) mg/dl. PMID- 16373027 TI - [C-reactive protein in the era of molecular medicine]. PMID- 16373026 TI - [Carotid ultrasound in the assessment of preclinical atherosclerosis. Distribution of intima-media thickness values and plaque frequency in a Spanish community cohort]. AB - BACKGROUND AND OBJECTIVE: High-resolution B-mode ultrasound measurements of carotid intima-media thickness (IMT) and determination of plaque presence are useful to assess preclinical atherosclerosis. Normal IMT values have not been reported in Spanish subjects. Our aim was to define normality data of carotid ultrasound by sex and age. SUBJECTS AND METHODS: We studied 250 healthy, normolipidemic subjects, 125 men and 125 women, with mean age 49 years (range, 20 81). We assessed cardiovascular risk factors and performed ultrasound determination of mean and maximum IMT in the far wall of the common carotid artery, plaque occurrence, and maximum plaque height. RESULTS: Reference values for carotid IMT, expressed as 25th, 50th, and 75th percentiles by sex and 5 age groups, have been obtained. The 50th percentiles of mean IMT ranged from 0.50 to 0.74 mm in men in the age groups 35 years or younger and 65 years or older, respectively. For women, corresponding IMT values ranged from 0.40 to 0.65 mm. IMT was strongly related (p < 0.001) to age, both in men (r = 0.57) and women (r = 0.61). From the regression equations, the estimated yearly increase in IMT was 0.005 mm in men and 0.007 mm in women. More than 50% of men aged 55 years and older, and of women aged 65 years and older, had carotid plaques. CONCLUSIONS: Both IMT and plaque frequency were associated with age in men and women. Carotid IMT values in a Spanish community cohort were lower than those reported for countries with higher cardiovascular risk, such as Northern European countries and the US. PMID- 16373028 TI - [Fibromyalgia]. PMID- 16373029 TI - [Neuraminidase inhibitors and their potential use in the avian influenza pandemic]. PMID- 16373030 TI - [Systematic review of pharmacologic treatment in fibromyalgia]. PMID- 16373031 TI - [Vitamin D and bone health: is there a need to review supplementation in osteoporosis risk population?]. AB - Vitamin D is an essential hormone for achieving an optimal bone physiology. There is no universal consensus nowadays on the definition of hypovitaminosis D and cut off values have been refined in the last years. The aim of this review is to analyze vitamin D deficiency among osteoporosis risk populations, including elderly and postmenopausal women, in Spain and other countries. We also review vitamin D supplementation: current clinical guidelines, last clinical studies, safety, and prescription schemes. PMID- 16373032 TI - [Mortality among patients admitted to hospitals on weekends]. PMID- 16373033 TI - [Geographical distribution of HFE C282Y and H63D mutation in Spain]. PMID- 16373034 TI - [Factors influencing plasma concentrations of C-reactive protein in patients with type 2 diabetes mellitus]. PMID- 16373035 TI - [Erythropoietin to treat anemia in cancer patients]. PMID- 16373036 TI - [Association and control of cardiovascular risk factors in the primary health care setting]. PMID- 16373037 TI - [Is homocysteine an independent risk factor for cardiovascular disease?]. PMID- 16373038 TI - [Transferring to the practice the results of the clinical trials with statins: from the relative risk to the absolute risk]. PMID- 16373039 TI - [New directions in the diagnosis of sleep apnea-hypopnea syndrome]. PMID- 16373040 TI - [Comparison between the 1993 and 2002 Guidelines of the Spanish Society of Pulmonology and Thoracic Surgery (SEPAR) for identifying respiratory events in polysomnography tests]. AB - OBJECTIVE: To compare the results of applying both the 1993 and 2002 guidelines of the Spanish Society of Pulmonology and Thoracic Surgery (SEPAR) to identify respiratory events during nighttime polysomnography tests. PATIENTS AND METHODS: One hundred twenty consecutive patients with medium to high suspicion of sleep apnea-hypopnea syndrome (SAHS) were included in the study. The 1993 guidelines recommended the use of a thermistor and the evaluation of only apneas and hypopneas. The 2002 guidelines, on the other hand, recommended the use of a thermistor, nasal pressure cannula, and thoracoabdominal bands so that respiratory effort related to arousals could be studied along with apneas and hypopneas. In our study we did not use an esophageal pressure catheter. We calculated the apnea index, hypopnea index, and apnea-hypopnea index (AHI) and determined the number of patients who would be diagnosed with SAHS (AH I > or =10) and the number for whom initiation of continuous positive airway pressure treatment would be recommended (AHI > or =30) according to the 2 sets of guidelines. RESULTS: Polysomnographic tests were valid for 118 of the 120 patients (80% men). The mean (SD) age was 51 (11.6) years and the mean body mass index 31.2 (4.3). Using the 1993 guidelines, the AHI was less than 10 in 25 patients, between 20 and 29 in 38, and 30 or more in 50. In the group overall, mean apnea and hypopnea indices and AHI were all significantly higher with the 2002 guidelines than with the 1993 criteria. With the 1993 criteria, the mean AHI was 33.16 and with 2002 criteria, 45.02 (P<.05). Sixty-four percent of the studies considered normal according to the 1993 SEPAR guidelines were considered apneic according to the 2002 guidelines. Of the patients considered not to need continuous positive airway pressure according to the 1993 SEPAR guidelines, 47.61% did need therapy according to the 2002 guidelines. CONCLUSIONS: There are significant differences in AHI, and in both apnea and hypopnea indices depending on whether the 1993 or the 2002 SEPAR guidelines are applied. PMID- 16373041 TI - [Smoking prevention intervention among secondary school students in the Spanish province of Malaga]. AB - OBJECTIVE: To assess the prevalence of smoking among 13- to 18-year-old students, analyze smoking-related factors in this population, and design an intervention program to reduce smoking. POPULATION AND METHODS: A 6-month smoking prevention intervention was designed for students at a public secondary school in the Spanish province of Malaga. The objective of the intervention was to inform students of the gravity of smoking and its deleterious effects on health. The efficacy of the intervention was subsequently assessed. RESULTS: Of the 337 students who completed the questionnaire, 27% declared they were smokers. Smoking prevalence was significantly higher among girls (36.1%) than boys (18.1%) (P<.001) and progressively increased with age. Initiation occurred at 13 to 14 years of age. The most common reason for starting was to try something new (52.2% of students). The greatest risk factor in the family environment was having a smoking sibling. After the program, 78% of smokers admitted that the intervention had not affected their smoking but had made them more aware of its detrimental effects, and 66.8% planned to quit in the future. CONCLUSIONS: The study shows the high prevalence of smoking among adolescents, particularly among girls. It also shows that health education increases understanding of the problems related to smoking, causing changes of attitude with regard to whether they will smoke in the future. PMID- 16373042 TI - [Geographic variation in the prevalence of asthma symptoms in Spanish children and adolescents. International Study of Asthma and Allergies in Childhood (ISAAC) Phase 3, Spain]. AB - OBJECTIVE: To analyze geographic variations in the prevalence of symptoms related to asthma in Spanish children and adolescents. POPULATION AND METHODS: In 2001 and 2002, the Spanish arm of the International Study of Asthma and Allergies in Childhood (ISAAC) Phase 3 collected information on 28 445 children in the age bracket of 6-7 years in 10 metropolitan areas (A Coruna, Asturias, Barcelona, Bilbao, Cartagena, Castellon, Madrid, Pamplona, San Sebastian, and Valencia) and on 31 257 adolescents in the bracket 13-14 years in 11 areas (the previously named areas plus Valladolid). An asthma symptom questionnaire was filled in by parents or the adolescents themselves. Differences in symptoms between geographic areas were analyzed by fitting a logistic regression model. The relationship between symptoms and age was analyzed by linear correlation. RESULTS: The prevalence of recent wheezing (last 12 months) ranged from 7.1% to 12.9% among 6 7-year-olds and from 7.1% to 15.3% among the 13-14-year-olds. The greatest risk of recent wheezing was observed for children in A Coruna (odds ratio [OR] =1.96 in comparison with the area of lowest prevalence; 95% confidence interval [CI], 1.65-2.33) and Bilbao (OR=1.83; 95% CI, 1.54-2.18) and for adolescents in A Coruna (OR=2.38; 95% CI, 2.04-2.79) and Asturias (OR=2.37; 95% CI, 2.03-2.77). A strong correlation (r=0.72) was observed between the prevalence of recent wheezing and age in each of the geographic areas. CONCLUSIONS: Considerable geographic variation in the prevalence of asthma symptoms can be seen in Spain even among young children. Symptoms are more frequent in children and adolescents who live on the Spain s northern Atlantic coast. PMID- 16373043 TI - [Variability in walk test conditions in pulmonary rehabilitation programs in Latin America and on the Iberian peninsula]. AB - OBJECTIVE: 1) To determine the frequency of use of the 6-minute walk test in pulmonary rehabilitation programs in Latin America and on the Iberian Peninsula; 2) to identify how the test is performed and possible variations from center to center. MATERIALS AND METHODS: A questionnaire was sent to 55 pulmonary rehabilitation centers in Latin America, Portugal, and Spain. RESULTS: Forty-nine (89.1%) centers answered the questionnaire. Forty-seven (95.9%) perform a walk test lasting 6 minutes; 22 (46.8%) take the longest distance of 2 tests as the real one; and 35 (74.5%) carry out the test in a corridor. The course distance ranged from 17 to 90 meters, but in 21 (44.7%) centers, the corridor was between 17 and 30 meters long. In 29 (61.7%) centers, the patients are routinely informed about the time that had elapsed during the test. Verbal encouragement is used in 44 (93.6%) centers. Thirty-eight (80.8%) use supplemental oxygen when a patient needs it. An increase in absolute values in the distance covered is used as a parameter indicating improvement at 21 (46.7%) sites while at 15 (33.3%) other centers the percentage increase is taken as a measure of improvement. CONCLUSIONS: The 6-minute walk test is widely used for the evaluation of the exercise capacity at the pulmonary rehabilitation centers of Latin America and the Iberian Peninsula. However, there is great variability in the way the test is performed. PMID- 16373044 TI - [Scientific impact of the Asthma Awareness Year, 2003: analysis of publications in Archivos de Bronconeumologia]. PMID- 16373045 TI - [Guidelines for the evaluation of surgical risk in bronchogenic carcinoma]. PMID- 16373046 TI - [Description of a new procedure for fiberoptic bronchoscopy during noninvasive ventilation through a nasal mask in patients with acute respiratory failure]. AB - A new method is described for performing oral fiberoptic bronchoscopy during noninvasive ventilation through the nose. The technique was successfully applied in 2 patients suffering from acute respiratory failure. The bronchoscope was inserted through a glove finger fitted into a mouth guard. The system works as a valve and does not affect performance of the bronchoscopy procedure or the pressures administered during noninvasive ventilation. We conclude that the procedure has potential advantages over bronchoscopy through the nose and face masks or helmets, particularly for the management of secretions or in special clinical circumstances (hemoptysis or presence of foreign bodies). This method can be used to substitute for or complement other bronchoscopy techniques performed with other interfaces. PMID- 16373047 TI - [Acute pneumonitis and adult respiratory distress syndrome after subcutaneous injection of liquid silicone]. PMID- 16373048 TI - [Malignant pleural mesothelioma: further options]. PMID- 16373049 TI - Clinical trials: bad combinations or bad designs. PMID- 16373050 TI - Cytoreductive surgery and intraoperative intraperitoneal hyperthermic chemotherapy in patients with peritoneal carcinomatosis of colorectal origin. AB - The median survival in patients with peritoneal carcinomatosis from colorectal adenocarcinoma is,with conventional approaches, only about six months. Combined treatment consisting of maxi-mum cytoreductive surgery plus intraoperative intraperitoneal hyperthermic chemotherapy has been shown, albeit in small non comparative series, to increase disease-free survival and overall survival, compared with previous series. Further, a randomized trial has demonstrated better results (a median survival of 22.4 months) with cytoreduction plus intraperitoneal chemotherapy compared with conventional chemotherapy. Technical considerations, infrastructure requirements and possible complications imply specialized centres and staff. Surgery consists of peritonectomy of affected areas and fulguration of all macroscopic lesions. Intraperitoneal chemotherapy must reach all parts of the peritoneal cavity and the temperature of the hyperthermic procedure must be maintained between 42-44 degrees C. Three prognostic factors associated with this procedure are: pathologic tumour grade, peritoneal carcinomatosis index, and cytoreductive surgery grade. PMID- 16373051 TI - Interdisciplinary clinical evaluation of 58 patients with lumbar-vertebral metastases from cervico-uterine cancer. AB - INTRODUCTION: Metastases in the vertebrae of patients with cervical cancer (CeCa) can be difficult to diagnose, and the treatment is palliative in many cases. OBJECTIVES: The aim of this study was to assess the ti-me required for diagnosis, the lesion's locoregional extent and the therapeutic schemes applied, in a retrospective series of 58 patients with CeCa and with lumbar spinal metastases. METHODS: The cases were studied using an updated interdisciplinary analysis to determine the clinical and radiological variables. This study evaluated the site and extent of bone lesions and correlated these variables with instability of the spine and cord compression. RESULTS: The diagnosis of vertebrae metastases of Ce Ca required more than 3 months in most cases. Lumbar vertebrae L4 and L5 and specifically the vertebral body were the most-frequently affected si-tes. Systemic and/or extra-compartmental-extended metastases (MosV4) were observed in 44/58 patients. Radiotherapy was the only option in this group and the palliative effect achieved was minimal, or null. In 14/58 patients there was intra compartmental-extended (MosV2) and extra-compartmental limited (MosV3) single vertebral metastases and the 3 different treatment schemes were administered. In the cases treated with marginal resection of metastases, vertebroplasty plus adjuvant radiotherapy achieved significant palliative effect. CONCLUSIONS: In the present series of patients, the diagnosis of metastases of the lumbar vertebrae was late, and the disease was advanced. The results obtained with radiotherapy in advanced stage disease did not improve the quality of life of patients. Metastasectomy was the therapeutic scheme in cases with intermediate stage disease and was the basis of the integrated treatment We believe that it is necessary to shorten the diagnostic time and to apply a staging system for vertebral metastases so that appropriate individualised selection of interdisciplinary treatment would be facilitated. PMID- 16373052 TI - [Effectiveness of 5-fluoruracil and vinorelbine in patients who had received multi-treatments for metastatic breast cancer]. AB - PURPOSE: In this study we evaluated the activity and toxicity of a combination of 5-fluorouracil continuous infusion and vinorelbine as second or third line chemotherapy in metastatic breast cancer (MBC). PATIENTS AN METHODS: A total 24 patients who had received doxorrubicin and/or paclitaxel were included in this study. The regimen consisted in 5-fluorouracil 1 gr/m(2) BSA continuous infusion for 3 days and vinorelbine 30 mg/m(2) intravenous (IV) on day 1. The cycles were repeated every 21 days for 6 cycles. RESULTS: Objective responses were recorded in 37.5% (12.5% complete remission). The median disease-free survival was calculated 6.33 +/- 8.12 months (CI 95% of 3.43 months). Toxicity was observed in 12.5% of the patients and no treatment related deaths were recorded. CONCLUSION: The combination of 5-fluorouracil/vinorelbine at the dose administered is an effective regime in patients with MBC, with low toxicity and cost. PMID- 16373053 TI - [Fractionated stereotactic-guided radiotherapy in the treatment of pituitary adenomas]. AB - AIMS: To evaluate the survival rates, prognostic factors and adverse events in patients with pituitary adenomas following fractionated stereotactic-guided radiotherapy (FSRT). MATERIAL AND METHODS: Fifty-six patients with pituitary adenomas were treated with FSRT; 23 patients (41.1%) had primary adenomas, 33 had recurrent disease; 24 (42.9%) with non-functional and 32 (57.1%) with functional adenomas. Using conventional fractionation, median total dose administered was 54 Gy (range: 24-56 Gy). RESULTS: The median follow-up was 51 months (range: 9-102) and, at the time of analysis, 49 patients were alive and disease-free, 1 patient was alive with reduced visual acuity and biochemical indications of recurrence, 2 patients had died from the disease and 1 patient had died from unrelated causes. Overall survival was 94% (50/53) and overall local tumour control was 92% (49/53). Univariate analysis indicated hormonal secretion (ACTH) and previous radiotherapy as being statistically significant. Fourteen patients (25%) had minor side-effects during treatment and 3 patients (5.4%) had late-onset events; 2 with optical neuropathy (both patients had other relevant co-existing diseases) and 1 patient had brain necrosis (re-irradiation). CONCLUSION: Fractionated stereotactic-guided radiotherapy is an effective modality for the treatment of pituitary adenomas. Care is required in patients with co-morbidities and/or previously-irradiated recurrent tumour so as to minimise late-onset secondary effects. PMID- 16373054 TI - [Breast metastases]. AB - Metastases in the breast resulting from extra-mammary tumours are infrequent, and the prognosis is poor. The main challenge in diagnosis is to differentiate between this condition and that of primary breast cancer so as to avoid unnecessary surgery. We present two cases that we encountered recently. PMID- 16373055 TI - Gastrointestinal bleeding as the first manifestation of a burned-out tumour of the testis. AB - We discuss the clinical presentation and course of the disease of a 25-year-old male who had gastrointestinal (GIT) symptoms secondary to retroperitoneal lymph node proliferation of a germ-cell tumour of the testis. The pathology evaluation of the orchiectomy specimen classified it as a burned-out tumour of the testis, given the lack of tumour elements and the presence of typical scarring tissue. Biological is-sues leading to tumour regression are discussed, as well. PMID- 16373056 TI - Lumbar region intra-spinal primitive neuroectodermal tumour (PNET) combined with neurofibromatosis type 1. AB - Primitive neuroectodermal tumours (PNET) are aggressive neoplasias that are diagnosed, usually, in infancy. Their appearance in adulthood is rare and, exceptionally, in association with neurofibromatosis type I (NF-1). We present a case of a 37 year-old man with NF-1 combined with PNET in the intra-arachidial lumbar region. Diagnosis was by Nuclear Magnetic Resonance (NMR) and biopsy of soft tissue mass which showed a PNET with undifferentiated round cells and immunohistochemically positive for CD99, neurone-specific enolase, synaptophys in and LEU-7. Surgery was performed with spine decompression and resection of 80% of the tumour, with symptoms improvement. Radiotherapy was administered on the lumbosacral column, but only up to 30 Gy because of severe actinic enteritis and pan-cytopenia grade III. Six months later, the patient was hospitalized with deterioration in his overall clinical status with multi-organ involvement. The patient died and an autopsy was performed. The initial treatment of the PNET is surgery and, if possible, the radical extirpation of the tumour. Administration of radiotherapy and chemotherapy appears to increase survival. We comment on the clinical, histological, cytological and immunohistochemical aspects together with a review of the literature. To the best of our knowledge this is the first documentation of such a case. PMID- 16373057 TI - [Hypoxia: a therapeutic target]. PMID- 16373059 TI - Once-weekly dose of epoetinum alfa in cancer patients with anemia receiving radiotherapy. AB - INTRODUCTION: Anaemia is present in 30%-90% of all patients with cancer, and its origin is multifactorial. Human recombinant erythropoietin has been shown to be useful in treating anemia in patients with cancer. The aim of this study was to evaluate the effectiveness of treatment of anaemia with epoetin alfa(EPO) given as a single weekly dose, and its repercussions on quality of life (QoL). MATERIALS AND METHODS: From January to October 2002, a total of 139 patients referred to our service for radiotherapy (RT) had anemia and received treatment with EPO as a single weekly dose of 40,000 IU subcutaneously, with oral iron supplement If haemoglobin (Hb) values after 1 month of treatment did not increase by > or =1 g/dl, the dose was increased to 60,000 IU/week. Treatment with EPO ended when Hb values reached > or =14 g/dl or one month after the end of RT regardless of Hb values. QoL was evaluated with the Functional Assessment of Cancer Therapy-Anaemia subscale (FACT-An) and the Cancer Linear Analogue Scale (CLAS). RESULTS: Mean Hb at the start of treatment with EPO was 11.49 +/- 1.08 g/dl, and the mean value at the end of treatment was 14.52 +/- 1.41 g/dl (p < 0.001). The mean increase in Hb was 2.97 +/- 1.65 g/dl. Mean duration of treatment was 7.13 +/- 2.91 weeks. In 11 patients (7.9%) the dose was increased after 4 weeks. In 84 patients (60.4%) EPO treatment was implemented before the commencing of RT. Mean Hb values in this group was 11.34 +/- 1.11 g/dl at the start of EPO treatment, 12.69 +/- 1.56 g/dl at the start of RT, 13.96 +1.54 g/dl at the end of RT and 14.68 +/- 1.3 g/dl at the end of EPO treatment (p < 0.001). In 55 patients(39.6%) anaemia developed during RT and, therefore, EPO treatment was implemented after commencing of RT. In this group the mean Hb values were 12.29 +/- 1.6 g/dl at the start of RT, 11.72 +/- 1.01 g/dl at the start of EPO treatment, 13.97 +/- 1.53 g/dl at the end of RT and 14.28 +/- 1.54 g/dl at the end of EPO treatment (p < 0.001). Hemoglobin levels at the start of EPO were lower in patients who commenced EPO before RT (p < 0.05). In 60 patients who received combined RT and chemotherapy, mean Hb values were 11.42 +/- 1.16 g/dl at the start of EPO and 13.98 +1.55 g/dl at the end of EPO (p < 0.005). In 75 patients who had received RT alone, the mean Hb values was 11.53 +/- 1.05 g/dl at the start of EPO and 14.98 +/- 1.17 g/dl at the end of treatment (p < 0.001). Patients treated with RT alone had higher Hb levels at the end of RT and at the end of EPO treatment than did patients who had received combined treatment(p < 0.005). The duration of EPO treatment was shorter in the group treated with RT alone than in the combined treatment group (6.41 +/- 2.99 weeks versus 7.96 +/- 2.67 weeks; p < 0.005). No significant differences were observed in FACT-An and CLAS scores at the beginning and the end of the study. CONCLUSIONS: Treatment with epoetin alfa as a single weekly dose significantly increased Hb levels in patients with cancer who were undergoing radiotherapy. The response was greater in patients treated with radiotherapy alone than in those receiving combined therapy. The duration of EPO treatment was shorter in the group treated with radiotherapy alone than in the combined treatment group. PMID- 16373058 TI - Double minutes, cytogenetic equivalents of gene amplification, in human neoplasia - a review. AB - Double minutes are tiny spherical chromatin bodies of a few mega-base pairs of size which are found occasionally in hematopoietic neoplasia and more or less often in human solid tumors. They have been associated with worse prognosis and poor outcome of the malignancies where present. With the beginning era of molecular cytogenetics they could be defined as cytogenetic equivalents of amplified DNA sequences. The identification of involved chromosomal segments and their molecular nature led to the development of molecular genetic techniques for a rapid and reliable detection of prognostically important oncogene amplifications in human tumors and,as a consequence, to gene-targeted therapy. PMID- 16373060 TI - A phase II trial of gemcitabine and weekly high-dose 5-fluorouracil in a 48-hour continuous-infusion schedule in patients with advanced pancreatic carcinoma. A study of the Spanish Cooperative Group for Gastrointestinal Tumour Therapy (TTD). AB - AIM: A multi-centred, open-labelled, phase 11 study containing 46 patients was conducted to evaluate the clinical benefit of gemcitabine (1,400 mg/m(2)) combined with 5-FU (3 g/m(2)) in a 48 h continuous infusion (CI). METHODS: Both drugs were administered on days 1, 8 and 15 of every 4 week cycle in chemotherapy naive patients with locally advanced unresectable metastatic pancreatic carcinoma. The minimum follow-up was 6 months. RESULTS: Clinical benefit response was the primary endpoint and this was achieved by 24.4% of the patients. Quality of life (QoL) improved in 16.6% of patients. Objective response was observed in 7% of the patients. The median progression-free survival (PFS) was 14.4 weeks and the median overall survival (OS) time was 22.7 weeks. One-year survival was 25%. The most frequent grade 3-4 toxicities were neutropenia (45%), mucositis (7.5%) and hyperbilirubinaemia (10.5%). CONCLUSIONS: This schedule was not superior in terms of clinical benefit, response rate, PFS and OS than standard gemcitabine treatment PMID- 16373061 TI - Psychosocial effect of mastectomy versus conservative surgery in patients with early breast cancer. AB - PURPOSE: To compare the quality-of-life (QoL) and psychosocial changes in a group of patients with early breast cancer who underwent conservative surgery (BCS) or modified radical mastectomy(MRM). METHODS: Self-administered questionnaire assessing body image perception, social habits, sexual attraction and self consciousness with relatives/friends, was randomly assigned to 125 patients (61 BCS, 64 MRM; aged 53 +/- 8 and 50 + 9 years, respectively, p = NS). RESULTS: MRM patients reported a significantly higher frequency of changes in body image perception and other related social behaviour such as avoiding going to the beach or using low-cut clothes, and reticence with friends. Conversely, no differences were found regarding sexuality, denial of the disease by the husband/partner, or concealing the disease from family members. Also, no significant differences were found between patients above and below the age of 50 years, for all variables studied after adjustment for surgical procedure. CONCLUSIONS: Modified radical mastectomy has a negative effect on body image perception and in social behaviour patterns of patients and with a concomitant decrease in QoL. The sexuality of the patient is not significantly affected. PMID- 16373062 TI - [Molecular protocol for HER2/neu analysis in breast carcinoma]. AB - BACKGROUND: The HER2/neu proto-oncogene is frequently over-expressed in breast cancer and serves as a biological target for trastuzumab therapy. However, there is no consensus regarding the technical aspects to be used to define HER2/neu status in clinical practice. METHODS: The present study was conducted to address this critical issue by prospectively analysing a large cohort of breast cancer patients (n = 222) and using a variety of methods. To define HER2/neu expression, detection of its encoded protein (p185) was performed by comparative immunohistochemical (IHC) analysis using two mouse monoclonal antibodies (mAb CB11 and mAb TAB250). To assess HER2/neu gene amplification, fluorescent in situ hybridisation (FISH) assays with gene-specific probes were conducted. All procedures were applied to de-paraffinised tissue sections of breast tumour samples. RESULTS: Results showed that mAb CB11 had increased sensitivity and specificity (62.5% and 93.4%, respectively) compared to mAb TAB250 (40% and 76.4%, respectively) in defining HER2/neu amplification. We conclude that HER2/neu measurement by IHC using mAb CB11 is an appropriate strategy which provides a high negative predictive value (95.5%) for HER2/neu amplification in cases with low or undetectable p185 expression. Conversely, mAb CB11 has a high positive predictive value (96.2%) for HER2/neu amplification in cases with p185 overexpression. However, cases with moderate p185 expression need to be considered as inconclusive. In such cases, it is necessary to use FISH measurement to evaluate HER2/neu amplification. It is also advisable to conduct FISH if there is discordance between p185 expression and the histopathology classification of the lesion, or molecular profile of the tumour. Finally, even though the false positive rate of IHC assay is <5%, the toxicity and cost of trastuzumab therapy suggest that FISH be used systematically prior to implementation of treatment. CONCLUSION: We suggest the use of a molecular protocol for HER2/neu analysis in this type of tumor. PMID- 16373063 TI - Pontine metastases as a cause of dysphagia in lung carcinoma. AB - Dysphagia is an unusual symptom in the clinical course of lung carcinoma. When it appears, it is necessary to differentiate between regional dissemination, drug toxicity, opportunistic infection and, most rarely, metastatic dissemination to the brain stem. Magnetic resonance imaging (MRI) is the best diagnostic option to exclude this last possibility. We present a male patient with progressive dysphagia 15 months after the diagnosis of an oat-cell lung carcinoma. Cerebral MRI revealed a pontine lesion, probably of metastatic origin. PMID- 16373064 TI - Interstitial pneumonitis after oxaliplatin treatment in colorectal cancer. AB - Progressive respiratory failure developed in a 68 year-old female who was treated with single-agent oxaliplatin for colorectal cancer. Only one cycle of 5 fluorouracil had been previously administered. Computed tomography of the chest showed lesions that suggested pulmonary fibrosis. There was an unfavourable response to treatment with corticosteroids, antimicrobial and antifungical agents. Lung biopsy findings were compatible with interstitial pneumonitis. The patient died 20 days after admission due to irreversible respiratory failure. This is the first case reported in the literature of interstitial pneumonitis related to single-agent oxaliplatin administration. PMID- 16373065 TI - Esophageal squamous cell carcinoma after liver transplantation. AB - The occurrence of an oesophageal squamous cell carcinoma following liver transplantation is very infrequent. Such an event has been related to a history of alcohol-induced cirrhosis, as in other squamous cell tumours of the oropharynx. We report the case of a 64-year-old male patient diagnosed as having oesophageal squamous cell carcinoma six years after having had a liver transplant due to alcohol-induced cirrhosis. The tumour was treated surgically and consisted of an Ivor-Lewis oesophagectomy. The patient is disease-free 17 months after surgery. A review of the cases reported in the literature indicated treatment with chemotherapy and radiation therapy, and with excision in some cases. Generally, despite aggressive treatment the prognosis is poor. PMID- 16373066 TI - A randomized, placebo-controlled trial of OROS methylphenidate in adults with attention-deficit/hyperactivity disorder. AB - BACKGROUND: The objective of this study was to evaluate the safety and efficacy of once-daily OROS methylphenidate (MPH) in the treatment of adults with DSM-IV attention-deficit/hyperactivity disorder (ADHD). METHODS: We conducted a randomized, 6-week, placebo-controlled, parallel-design study of OROS MPH in 141 adult subjects with DSM-IV ADHD, using standardized instruments for diagnosis. OROS MPH or placebo was initiated at 36 mg/day and titrated to optimal response, depending on efficacy and tolerability, up to 1.3 mg/kg/day. RESULTS: Treatment with OROS MPH was associated with clinically and statistically significant reductions in DSM-IV symptoms of inattention and hyperactivity/impulsivity relative to subjects treated with placebo. At endpoint, 66% of subjects (n = 44) receiving OROS MPH and 39% of subjects (n = 29) [corrected] receiving placebo attained our a priori definition of response of much or very much improved on the Clinical Global Impression-Improvement scale plus a >30% reduction in Adult ADHD Investigator System Report Scale score. OROS MPH was associated with small but statistically significant increases in systolic blood pressure (3.5 +/- 11.8 mm Hg), diastolic blood pressure (4.0 +/- 8.5 mm Hg), and heart rate (4.5 +/- 10.5 bpm). CONCLUSIONS: These results show that treatment with OROS MPH in daily doses of up to 1.3 mg/kg/day was effective in the treatment of adults with ADHD. Because of the potential for increases in blood pressure and heart rate, subjects receiving treatment with MPH should be monitored for changes in blood pressure parameters during treatment. PMID- 16373067 TI - Opposing influence of basolateral amygdala and footshock stimulation on neurons of the central amygdala. AB - BACKGROUND: The basolateral complex (BLA) and the central nucleus of the amygdala (CeA) are believed to mediate the expression of affective responses. After affective learning, conditioned stimulus-related information is thought to be conveyed from the BLA to the CeA; the medial CeA (Cem), in turn, projects to hypothalamic and brainstem structures involved with induction of affective responses. Although the conditioned stimulus and unconditioned stimulus both evoke affective responses, the precise response often differs. It is unknown whether this difference is represented by distinct activity patterns of single Cem neurons. Furthermore, the nature of the interaction between the BLA and Cem is unknown. METHODS: Using in vivo extracellular and intracellular recordings, we examined how the BLA affects the Cem and compared this with effects induced by footshock (unconditioned stimulus) in the same neurons. RESULTS: Our results demonstrate that, contrary to conventional views, BLA stimulation primarily inhibits Cem neurons by a polysynaptic circuit, and show that single Cem neurons respond to both BLA input and footshock in an opposite manner. CONCLUSIONS: These results demonstrate the predominantly inhibitory nature of the BLA-Cem interaction. These data further demonstrate the distinct cellular events that might lead to differential modulation of conditioned and unconditioned affective responses. PMID- 16373068 TI - High-affinity states of human brain dopamine D2/3 receptors imaged by the agonist [11C]-(+)-PHNO. AB - BACKGROUND: The high-affinity states of dopamine D2-receptors (D2(high)) are postulated to be functionally responsible for signal transduction. At present, no useful in vivo method exists to selectively measure D2(high) in humans, as current D2 radioligands for positron emission tomography (PET) are either not D2 selective or do not differentiate between D2 high- and low-affinity states. METHODS: The D2-agonist (+)-PHNO [(+)4-propyl-9-hydroxynaphthoxazine] was labeled with carbon-11 and studied with PET. Eight [11C]-(+)-PHNO scans were acquired in four healthy volunteers. RESULTS: We observed greatest [11C]-(+)-PHNO accumulation in caudate, putamen, and globus pallidus [binding potentials (BPs): 3.00 +/- .4, 3.10 +/- .2, and 4.17 +/- 1.2]. Small but detectable binding was identified in the substantia nigra/ventral tegmental area. Preliminary test retest data in two subjects suggests BP-estimates to be reliable. Pre-treatment with haloperidol reduced BPs in regions showing specific binding with no detectable changes in cerebellum. Parallel imaging with [11C]-raclopride showed substantial differences in the globus pallidus. CONCLUSIONS: [11C]-(+)-PHNO proved to be a D2/3-receptor agonist-radioligand with good brain uptake and favorable kinetics for PET in humans. [11C]-(+)-PHNO delineated D2/3-receptor rich brain regions with high signal-to-noise ratio. This is the first demonstration of a viable agonist-radioligand for D2 receptors in humans and opens the door for investigating D2(high) in health and disease. PMID- 16373069 TI - Biochemical characterization of the glutamate transport in Trypanosoma cruzi. AB - The role of amino acids in trypanosomatids goes beyond protein synthesis, involving processes such as differentiation, osmoregulation and energy metabolism. The availability of the amino acids involved in those functions depends, among other things, on their transport into the cell. Here we characterize a glutamate transporter from the human protozoan parasite Trypanosoma cruzi. Kinetic data show a single saturable system with a Km of 0.30 mM and a maximum velocity of 98.34 pmoles min(-1) per 2 x 10(7) cells for epimastigotes and 20 pmoles min(-1) per 2 x 10(7) cells for trypomastigotes. Transport was not affected by parasite nutrient starvation for up to 3h. Aspartate, alanine, glutamine, asparagine, methionine, oxaloacetate and alpha ketoglutarate competed with the substrate in 10-fold excess concentrations. Glutamate uptake was strongly dependent on pH, but not on Na+ or K+ concentrations in the extracellular medium. These data were consistent with the sensitivity of the system to the H+ ionophore carbonyl cyanide p trifluoromethoxyphenylhydrazone, suggesting that transport is driven by H+ concentration gradient across the cytoplasmic membrane. The glutamate transport increased linearly with temperature in a range from 15 to 40 degrees C, allowing the calculation of an activation energy of 52.38 kJ/mol. PMID- 16373070 TI - Transmission of freshwater myxozoans during the asexual propagation of invertebrate hosts. AB - The phylum Myxozoa contains over 1350 species almost all of which are considered to be obligate parasites of aquatic animals. The phylum is composed of two classes, the Myxosporea and the Malacosporea, species of which are important pathogens responsible for severe economic losses in cultured fisheries. The life cycles of freshwater Myxozoa are believed to involve horizontal, indirect transmission, involving an invertebrate (oligochaetes or bryozoans) and a vertebrate host (fish or amphibians). Here, we describe myxozoan propagation through the fragmentation of invertebrate hosts to form new infected individuals. The two hosts examined are an oligochaete Lumbriculus variegatus infected with an unidentified myxosporean (Triactinomyxon sp.) and the bryozoan Fredericella sultana infected with the malacosporean Tetracapsuloides bryosalmonae which causes proliferative kidney disease, a major constraint of the European rainbow trout industry. Such intra-clonal propagation is a novel form of vertical transmission that is likely to be widespread within the Myxozoa and could form an important method by which some of these parasites maintain and proliferate within the aquatic environment. PMID- 16373071 TI - Solution structure of a two-repeat fragment of major vault protein. AB - Major vault protein (MVP) is the main constituent of vaults, large ribonucleoprotein particles implicated in resistance to cancer therapy and correlated with poor survival prognosis. Here, we report the structure of the main repeat element in human MVP. The approximately 55 amino acid residue MVP domain has a unique, novel fold that consists of a three-stranded antiparallel beta-sheet. The solution NMR structure of a two-domain fragment reveals the interdomain contacts and relative orientations of the two MVP domains. We use these results to model the assembly of 672 MVP domains from 96 MVP molecules into the ribs of the 13MDa vault structure. The unique features include a thin, skin like structure with polar residues on both the cytoplasmic and internal surface, and a pole-to-pole arrangement of MVP molecules. These studies provide a starting point for understanding the self-assembly of MVP into vaults and their interactions with other proteins. Chemical shift perturbation studies identified the binding site of vault poly(ADP-ribose) polymerase, another component of vault particles, indicating that MVP domains form a new class of interaction-mediating modules. PMID- 16373072 TI - Unlocking of the filamentous bacteriophage virion during infection is mediated by the C domain of pIII. AB - Protein III (pIII) of filamentous phage is required for both the beginning and the end of the phage life cycle. The infection starts by binding of the N terminal N2 and N1 domains to the primary and secondary host receptors, F pilus and TolA protein, respectively, whereas the life cycle terminates by the C terminal domain-mediated release of the membrane-anchored virion from the cell. It has been assumed that the role of the C-terminal domain of pIII in the infection is that of a tether for the receptor-binding domains N1N2 to the main body of the virion. In a poorly understood process that follows receptor binding, the virion disassembles as its protein(s) become integrated into the host inner membrane, resulting in the phage genome entry into the bacterial cytoplasm. To begin revealing the mechanism of this process, we showed that tethering the functional N1N2 receptor-binding domain to the virion via termination-incompetent C domain abolishes infection. This infection defect cannot be complemented by in trans supply of the functional C domain. Therefore, the C domain of pIII acts in concert with the receptor-binding domains to mediate the post receptor binding events in the infection. Based on these findings, we propose a model in which binding of the N1 domain to the periplasmic portion of TolA, the secondary receptor, triggers in cis a conformational change in the C domain, and that this change opens or unlocks the pIII end of the virion, allowing the entry phase of infection to proceed. To our knowledge, this is the first virus that uses the same protein domain both for the insertion into and release from the host membrane. PMID- 16373073 TI - Histopathological changes induced by the venom of the snake Vipera raddei (Armenian adder). AB - The venom of the Armenian adder (Vipera raddei Boettger, 1890) was tested for its ability to induce histopathological changes in rabbits after long-term (once every 6 days for 30 days) intramuscular injection of the venom (0.35 mg/kg approx. 0.5 LD 50), by light microscopic examination of some organs (liver, heart, kidney, adrenal, lung, spleen). V. raddei (Vr) venom induces changes including necrosis and edematous appearance with cellular infiltration and vacuolation. The injury of kidneys includes significant changes of the glomerular apparatus. Lung sections showed infiltration of erythrocytes within bronchial alveoli and dilation of bronchi. Liver sections showed vascular damage, significant quantities of hemosiderin and also presence of lymphohistiocyte elements. In the heart, Armenian adder venom produced slight disruption of the structure of cardiomyocytes, fibrosis and congestion of blood vessels. The histological structure of the spleen was not disturbed by the venom but there were lymphoblasts and numerous polynuclear macrophages within the central zone of follicules, which may indicate a development of moderate inflammatory reactions and activation of the complement system. Our results suggest that the venom of V. raddei induces moderate histopathological changes in vital organs. PMID- 16373074 TI - Lipoxygenase-derived eicosanoids are involved in the inhibitory effect of Crotalus durissus terrificus venom or crotoxin on rat macrophage phagocytosis. AB - Crotalus durissus terrificus snake venom and its major toxin, crotoxin or type II PLA2 subunit of this toxin, induce an inhibitory effect on spreading and phagocytosis in 2h incubated macrophages. The involvement of arachidonate-derived mediators on the inhibitory action of the venom or toxins on rat peritoneal macrophage phagocytosis was presently investigated. Peritoneal cells harvested from naive rats and incubated with the venom or toxins or harvested from the peritoneal cavity of rats pre-treated with the toxins were used. Zileuton, a 5 lipoxygenase inhibitor but not indomethacin, a cyclooxygenase inhibitor, given in vivo and in vitro abolished the inhibitory effect of venom or toxins on phagocytosis. Resident peritoneal macrophages incubated with the venom or toxins showed increased levels of prostaglandin E2 and lipoxin A4, with no change in leukotriene B4. These results suggest that lipoxygenase-derived eicosanoids are involved in the inhibitory effect of C.d. terrificus venom, crotoxin or PLA2 on macrophage phagocytosis. PMID- 16373075 TI - Molecular cloning of novel serine proteases and phospholipases A2 from green pit viper (Trimeresurus albolabris) venom gland cDNA library. AB - Green pit viper (Trimeresurus albolabris) is the most common venomous snake responsible for bites in Bangkok. It causes local edema and systemic hypofibrinogenemia resulted from the thrombin-like, as well as the fibrinolytic effects of the venom. However, the amino acid sequences of these venom proteins have never been reported. In this study, we have cloned five novel serine proteases from the Thai T. albolabris venom gland cDNA library. They were all closely homologous to the corresponding serine proteases from Chinese green viper (Trimeresurus stejnegeri), suggesting the evolutionary proximity of the two species. In addition, their functional activities could be deduced. There were predicted to be two thrombin-like enzymes (GPV-TL1 and GPV-TL-2), two isoforms of a fibrinogenolytic enzyme (albofibrase) and a plasminogen activator (GPV-PA), suggesting that defibrination syndrome in patients is a combination of these enzymatic effects. By multiple sequence alignment, no conserved residue or motif responsible for distinct functions of snake venom serine proteases could be observed. Moreover, one Lys 49 and one Asn 49 phospholipase A2 (PLA2) genes were cloned. Lys 49 PLA2 was predicted to devoid of catalytic activity, but showed a carboxy terminal cytotoxic region. No Asp 49 PLA2 was found in 150 clones screened. This explains the marked limb edema but no hemolysis in patients. These novel serine proteases have potentials to be therapeutic anti-thrombotic and thrombolytic agents in the future. PMID- 16373076 TI - Sex-based individual variation of snake venom proteome among eighteen Bothrops jararaca siblings. AB - Variation of venom proteome is relevant to basic research, to management of envenoming, and to studies on the evolution of poisonous snakes. In this study, we explored the venom proteomes of eighteen Bothrops jararaca specimens of a single litter born and raised in laboratory. Using electrophoretic techniques and various protocols for measuring the proteolytic activities of these venoms we have detected individual variability and highlighted sex-specific proteomic similarities and differences among sibling snakes. SDS-polyacrylamide gel electrophoresis under non-reducing conditions showed protein bands of approximately 100 kDa specific of male venoms. 2D-electrophoresis showed regions with varying spot complexity between pooled female and male venoms as well as spots that were gender specific. Gelatin zymography showed that female venoms contained proteinases of approximately 25 kDa absent from male venoms. Female venoms were more active than male venoms in degrading fibrinogen whereas on fibrin no significant differences were detected. Among various chromogenic peptide substrates tested, male venoms showed higher amidolytic activity than female venoms on D-Val-Leu-Lys-pNA and D-Phe-Pip-Arg-pNA. Taken together, these results show sex-based differences in the venom proteome of sibling snakes of a single litter raised under controlled conditions which seem to be genetically inherited and imposed by evolutionary forces. PMID- 16373077 TI - Regional variations in the distributions of small intestinal intraepithelial lymphocytes (IELs) in outbred laboratory mice (Mus musculus domesticus) and the inbred strain of mice established from Japanese fancy mice (Mus musculus molossinus). AB - Previously, we reported regional variations in small intestinal IELs of mice. In this study, we examined the regional variations of IELs in outbred laboratory mice (ddY: Mus musculus domesticus) and the inbred strain of mice established from Japanese fancy mice (JF1: Mus musculus molossinus). IELs were isolated from the proximal, middle and distal parts of the small intestine and analyzed by flow cytometry. The percentages of gammadelta T cells and alphabeta T cell subset of extrathymic origin were higher in the proximal part while the percentages of alphabeta T cell subset(s) of thymic origin were higher in the distal part in both ddY and JF1 mice. Such trends in regional variations of IELs were almost the same as those found in the inbred strains of laboratory mice in our previous reports. This strongly suggests that these regional variations of IELs may be common phenomena in Mus musculus species. PMID- 16373078 TI - Decommissioning of offshore oil and gas facilities: a comparative assessment of different scenarios. AB - A material and energy flow analysis, with corresponding financial flows, was carried out for different decommissioning scenarios for the different elements of an offshore oil and gas structure. A comparative assessment was made of the non financial (especially environmental) outcomes of the different scenarios, with the reference scenario being to leave all structures in situ, while other scenarios envisaged leaving them on the seabed or removing them to shore for recycling and disposal. The costs of each scenario, when compared with the reference scenario, give an implicit valuation of the non-financial outcomes (e.g. environmental improvements), should that scenario be adopted by society. The paper concludes that it is not clear that the removal of the topsides and jackets of large steel structures to shore, as currently required by regulations, is environmentally justified; that concrete structures should certainly be left in place; and that leaving footings, cuttings and pipelines in place, with subsequent monitoring, would also be justified unless very large values were placed by society on a clear seabed and trawling access. PMID- 16373079 TI - Respirometry-based evaluation of the fate and possible effects of antifreeze on activated sludge. AB - This paper focuses on environmental concerns regarding used antifreeze. It shows the results of laboratory respirometric tests conducted to evaluate the potential impact of antifreeze on activated sludge. In this first step of the study, only the components of virgin antifreeze were addressed, i.e. ethylene glycol, and antifoaming and corrosion inhibitory substances. The plateau resulting from oxygen uptake rate measurements of mixed liquor taken at the end of an aeration tank is used as a reference to evaluate the response of biomass to pulse injection of increasing dosages of new antifreeze, compared to that of formaldehyde, a known toxic compound. While 200 mg/L of formaldehyde steadily decreased the biomass activity (7 mg O2/Lh), there were no toxicity effects for doses up to 5000 mg/L of ethylene glycol from new antifreeze coolant (23 mg O2/Lh, compared to the initial 15 mg O2/Lh for the mixed liquor). It is estimated that ethylene glycol concentration in municipal wastewaters should be several order of magnitude lower then the 1000 mg/L shown to be tolerated by the biomass in this work, but at the present stage, the research does not provide enough information to recommend a definitive management scheme for antifreeze discharge. Many other management issues for used antifreeze must be addressed in future studies before any decision on tolerating sewer discharge in Mexico, as a transitory management option that will conciliate the actual practices with legality, can be made. PMID- 16373080 TI - Stereotactic linac-based radiosurgery in the treatment of cerebral arteriovenous malformations located deep, involving corpus callosum, motor cortex, or brainstem. AB - PURPOSE: To evaluate patient outcome and obliteration rates after radiosurgery (RS) for cerebral arteriovenous malformations (AVM) located deep, in the motor cortex or brainstem and those involving corpus callosum. METHODS AND MATERIALS: This analysis is based on 65 patients. AVM classification according to Spetzler Martin was 13 patients Grade 2, 39 Grade 3, 12 Grade 4, and 1 Grade 5. Median RS based AVM score was 1.69. Median single dose was 18 Gy. Mean treatment volume was 5.2 cc (range, 0.2-26.5 cc). Forty patients (62%) experienced intracranial hemorrhage before RS. Median follow-up was 3.0 years. RESULTS: Actuarial complete obliteration rate (CO) was 50% and 65% after 3 and 5 years, respectively. CO was significantly higher in AVM <3 cm (p < 0.02) and after doses >18 Gy (p < 0.009). Annual bleeding risk after RS was 4.7%, 3.4%, and 2.7% after 1, 2, and 3 years, respectively. AVM >3 cm (p < 0.01), AVM volume >4 cc (p < 0.009), and AVM score >1.5 (p < 0.02) showed a significant higher bleeding risk. Neurologic dysfunction improved, completely dissolved, or remained stable in 94% of patients. CONCLUSIONS: Surgically inaccessible AVM can be successfully treated using RS with acceptable obliteration rates and low risk for late morbidity. The risk of intracranial hemorrhage is reduced after RS and depends on RS-based AVM score. PMID- 16373081 TI - Spinal cord gliomas: A multi-institutional retrospective analysis. AB - PURPOSE: To determine the impact of postoperative radiation therapy (POXRT) on outcome in spinal cord gliomas. PATIENTS AND METHODS: Data from 242 patients were collected retrospectively from six institutions using a standardized data sheet. Pathology specimens, when available, were centrally reviewed. RESULTS: A total of 183 patients were analyzed: 82 received surgery alone as initial treatment, whereas 101 had surgery and POXRT. Demographic, diagnostic, and treatment factors were analyzed for impact on progression-free (PFS) and overall survival (OS). PFS in ependymoma patients was 74%, 60%, and 35% at 5, 10, 15 years, respectively, and was significantly influenced by treatment type, race, age, tumor grade, and type of surgery on univariate analysis, with age being the only significant factor on multivariate analysis (MVA) (p = 0.01). OS of ependymoma patients was 91%, 84%, and 75% at 5, 10, and 15 years, respectively, and was significantly influenced by both complete resection (p = 0.04) and age (p = 0.03) on MVA. In astrocytomas, PFS was 42%, 29%, and 15% at 5, 10, and 15 years, and was significantly influenced by POXRT in low- and intermediate-grade tumors on MVA (p = 0.02). OS at 5, 10, and 15 years was 59%, 53%, and 32%, respectively, and was significantly influenced by grade on MVA (p < 0.01). CONCLUSION: Postoperative radiation therapy reduced disease progression in low- and moderate-grade astrocytomas. In ependymomas, complete resection significantly influenced OS. PMID- 16373082 TI - Radiation pneumonitis following concurrent accelerated hyperfractionated radiotherapy and chemotherapy for limited-stage small-cell lung cancer: Dose volume histogram analysis and comparison with conventional chemoradiation. AB - PURPOSE: The aim of this study was twofold: to determine whether the dose-volume metrics are valuable in predicting radiation pneumonitis (RP) in small-cell lung cancer (SCLC) patients treated with accelerated hyperfractionated radiotherapy and chemotherapy (AHFRT + CT); and to clarify how AHFRT influences the risk of RP in comparison to conventional once-daily radiotherapy and chemotherapy (QDRT + CT). METHODS AND MATERIALS: Study subjects were 43 patients with SCLC treated with AHFRT + CT. Radiotherapy was delivered at 1.5 Gy/fraction (fr) twice daily to 45 Gy/30 fr/3 weeks. We analyzed the relation between RP incidence and several dosimetric factors. We also compared this series data with our previously published data from lung cancer patients treated with QDRT + CT. RESULTS: Radiation pneumonitis Grades 1, 2, and 3 were observed in 28 patients, 7 patients, and 1 patient, respectively. Univariate analysis revealed that the percentage of lung volume receiving more than 15 Gy, 20 Gy, and 30 Gy (V15, V20, V30) and normal tissue complication probability were of predictive value for the development of RP. The 12-month cumulative incidences of RP greater than Grade 2 were 0%, 7.1%, 25%, and 42.9% in patients with a V20 of < or =20%, 21-25%, 26 30%, and > or =31%, respectively. These incidences were lower than that of our patients treated with QDRT + CT. CONCLUSIONS: Dosimetric factors are valuable in predicting RP in SCLC patients treated with AHFRT + CT. Regarding the incidence of RP, AHFRT appears to have some advantage over QDRT. PMID- 16373083 TI - Brain responses to tonal changes in the first two years of life. AB - Maturation of auditory perceptual and discrimination process within the first two years of life is investigated in healthy infants by examining event-related potentials (ERPs). High-density EEG signals were recorded from the scalp monthly between 3 and 24 months of age. Two types of stimuli (100 vs. 100 Hz for standard stimuli; 100 vs. 300 Hz for deviant stimuli; occurrence rate: 85:15%) were presented using an oddball paradigm. Latencies and amplitudes were compared across development. The results showed that latencies of P150, N250, P350, and N450 components gradually decreased with increasing age. Amplitudes of the N250 and P350 components gradually increased and reached the maximum at 9 months, and then gradually decreased with the increase of age. Mismatch negativity was not obvious at 3 months of age, but was seen at 4-5 months and became robust after 6 months. Robust late positivity was recorded at all ages. These mismatch responses were noticeable in the frontal, central, and parietal areas, and the maximal MMN amplitude distribution gradually moved from the parietal area to the frontal area across the age range. Two important periods--one around 6 months and the other around 9 months are suggested in the maturation of auditory central system. Dynamical changes in the underlying source strengths and orientations may be principal contributors to ERP morphological changes in infants within the first 24 months. PMID- 16373084 TI - Chemoocclusion vs chemoperfusion for treatment of advanced hepatocellular carcinoma: a randomised trial. AB - AIMS: Transarterial chemoembolization (TACE) can be associated with considerable toxicity and treatment-associated mortality. Transient transarterial chemoocclusion (TACO) using degradable starch microspheres (DSM) has been proposed as a potentially safer alternative while maintaining anti-tumour efficiency. In a randomised phase II trial TACO was compared to transarterial chemoperfusion without DSM (TACP). METHODS: Seventy-four patients with advanced HCC were randomised to two treatment arms: (i) TACO (600-1200 mg DSM) and (ii) TACP. In both arms regional chemotherapy consisted of cisplatin (100 mg/m2) and doxorubicin (60 mg/m2). Both arms were corresponding in terms of age, gender, liver performance state, and tumour-stage. A maximum of six treatment cycles was applied in monthly intervals. Follow-up was performed in terms of tumour response, time to progression, survival and quality of life. RESULTS: Tumour response rates did not differ significantly between the two treatment arms, however, there was a tendency towards higher response rates in the TACO arm (TACO vs TACP): partial response: 26 vs 9%, stable disease: 41 vs 55%, progressive disease: 33 vs 36%. Time to tumour progression (32 vs 27 weeks), and overall survival (60 vs 69 weeks) were not significantly different. Grade 4 adverse events were rare in both arms and treatment-associated mortality was not observed. In addition, there was no significant difference in terms of quality of life under therapy (EORTC). CONCLUSION: TACO with DSM did not improve response or survival significantly compared to TACP in advanced non-resectable HCC. PMID- 16373085 TI - Temporal and spatial mRNA expression patterns of TGF-beta1, 2, 3 and TbetaRI, II, III in skeletal muscles of mdx mice. AB - To address potential regulatory roles of TGF-beta1 in muscle inflammation and fibrosis associated with dystrophin deficiency, we performed quantitative RT-PCR and in situ hybridization to characterize the temporal and spatial mRNA expression patterns of TGF-beta1 and other TGF-beta subfamily members, TGF-beta2 and TGF-beta3, as well as their receptors, in quadriceps and diaphragm muscles of mdx mice. TGF-beta1 mRNA was markedly upregulated in the endomysial inflammatory cells and regenerating fibers of mdx quadriceps and diaphragm, with the mRNA levels correlated with the degree of endomysial inflammation. Upregulation of TGF beta2, beta3, and their receptors was also appreciated but to a much lesser degree. While high levels of TGF-beta1 mRNA remained in the aging mdx quadriceps but not the diaphragm, progressive fibrosis only occurred in the diaphragm. Our data support a regulatory role for TGF-beta1 in muscle inflammation in mdx mice. It also suggests different susceptibility of quadriceps and diaphragm muscles to fibrosis induced by TGF-beta1 signaling pathway. PMID- 16373086 TI - Novel frameshift and splice site mutations in the neurotrophic tyrosine kinase receptor type 1 gene (NTRK1) associated with hereditary sensory neuropathy type IV. AB - Congenital insensitivity to pain with anhidrosis or hereditary sensory and autonomic neuropathy type IV (HSAN IV) is the first human genetic disorder implicated in the neurotrophin signal transduction pathway. HSAN IV is characterized by absence of reaction to noxious stimuli, recurrent episodes of fever, anhidrosis, self-mutilating behavior and often mental retardation. Mutations in the neurotrophic tyrosine kinase, receptor, type 1 (NTRK1) are associated with this disorder. Here we report four homozygous mutations, two frameshift (p.Gln626fsX6 and p.Gly181fsX58), one missense (p.Arg761Trp) and one splice site (c.359+5G>T) mutation in four HSAN IV patients. The splice site mutation caused skipping of exons 2 and 3 in patient's mRNA resulting in an in frame deletion of the second leucine-rich motif. NTRK1 mutations are only rarely reported in the European population. This report extends the spectrum of NTRK1 mutations observed in patients diagnosed with HSAN IV. PMID- 16373087 TI - Clinical, electrophysiological and molecular genetic studies in a family with X linked dominant Charcot-Marie-Tooth neuropathy presenting a novel mutation in GJB1 Promoter and a rare polymorphism in LITAF/SIMPLE. AB - Charcot-Marie-Tooth disease is a genetically heterogeneous group of neuropathies. In the demyelinating form of Charcot-Marie-Tooth disease with dominant inheritance, five genes have been incriminated: PMP22, MPZ, LITAF/SIMPLE, EGR2 (CMT1A to D), and GJB1 (CMTX). Here, we report clinical, electrophysiological and molecular genetic studies in a family with a Charcot-Marie-Tooth disease variable phenotype, ranging from asymptomatic to moderately affected. The absence of male to-male transmission as well as the results of systematic electrophysiological studies suggested a CMTX secondary to a GJB1 mutation. Screening for mutations in the coding regions of PMP22, MPZ, EGR2 and GJB1 was negative. We identified (1) a LITAF/SIMPLE substitution (T49M), absent in 1000 control chromosomes, but which was thought to be a polymorphism because of discrepancies of segregation when considering the results of electrophysiology; and (2) a novel substitution T>C in the P2 promoter of GJB1 at position -529, in the SOX10 binding site S2. The transmission of this second mutation was consistent with the electrophysiological data. We emphasise the role of electrophysiological studies that help to discriminate between asymptomatic subjects and that bring some additional valuable data to the genetic approach. PMID- 16373088 TI - Recognising F-response interference as a source of increased jitter in stimulated single fibre EMG. AB - OBJECTIVE: To determine the source of an abnormal pattern of latency shifts leading to falsely high jitters in single fibre electromyography (SFEMG). METHODS: We observed a sudden shortening of the latency to an individual single fibre spike component followed by a gradual return to baseline values during stimulation single fibre electromyography (SFEMG) of the facial muscle. The pattern could be reproduced in healthy controls. RESULTS: The sudden decrease in latency proved to follow an additional discharge of the muscle fibre, not due to the external stimulus. This additional discharge was identified as an F-response. CONCLUSIONS: The mechanism is thought to be a higher muscle fibre conduction velocity resulting from a temporary increase in stimulus frequency, in the form of an extra impulse along the muscle fibre represented by the F-response. SIGNIFICANCE: The typical abnormal pattern should be recognised because it can falsely increase the mean jitter. We advice to increase the time base to 50 ms if this pattern is observed and to exclude the affected potentials from jitter measurements. PMID- 16373089 TI - Restating the importance of bipolar recording in subcortical nuclei. PMID- 16373091 TI - Pseudo-iron deficiency in a French population living near high-voltage transmission lines: a dilemma for clinicians. PMID- 16373090 TI - Laparoscopic surgery. AB - The generic advantages of avoiding a large laparotomy incision are now well established. For the patient, a laparoscopic procedure is invariably less painful, and recovery and return to full normal activities is more rapid. There are also significant gains in short-term quality of life measures associated with the laparoscopic approach. For the surgeon, improved visualisation offers the opportunity of more precise and accurate surgery. These advantages are usually offset by longer operating times, the use of complex and expensive equipment, and the possibility of new types of complications and increased risk of standard operative morbidity. The aim of this chapter is to identify areas of general technique in which the risks associated with laparoscopic surgery can be minimised while retaining all the advantages of the approach. This is being achieved partly by improved and simplified instrumentation, partly by refinement in techniques, and partly by an increasing awareness of the potential pitfalls of the approach and by adopting strategies to avoid these problems. The majority of gynaecological procedures are already performed endoscopically and all gynaecologists who operate will need to become proficient in these techniques. This chapter outlines techniques for safe laparoscopic entry and safe bipolar diathermy techniques, and describes how to undertake some of the simpler laparoscopic procedures; the evidence supporting these approaches is also presented. Surgical proficiency in safe laparoscopic entry and laparoscopic tubal surgery should lead to the confidence to then undertake more complex procedures. PMID- 16373092 TI - Post-earthquake illness and disease after the Chi-Chi earthquake. PMID- 16373093 TI - EMS is a competitive sport. PMID- 16373095 TI - Get rid of this guy. PMID- 16373094 TI - Prepped & ready. PMID- 16373096 TI - Monsters at the gate: are you making people barf? PMID- 16373097 TI - The enemy of excellence. PMID- 16373100 TI - Katrina exposes known weaknesses. PMID- 16373102 TI - What did you do today? PMID- 16373104 TI - All that wheezes is not asthma. PMID- 16373109 TI - Trapped on urban islands. PMID- 16373110 TI - A straight-shooting leader. PMID- 16373111 TI - Dangerous care? PMID- 16373112 TI - Quality check. PMID- 16373113 TI - Teach airway, not tubes. PMID- 16373114 TI - Pen tricks. PMID- 16373116 TI - Time is brain: causes, imitators & prehospital measures of stroke. PMID- 16373117 TI - The Chaos of Katrina: EMS maintains composure in the midst of anarchy. PMID- 16373118 TI - When the rules change. PMID- 16373119 TI - Behind Katrina's eye with New Orleans EMS. PMID- 16373120 TI - Surviving the superdome. PMID- 16373121 TI - 72 hours: what rescuers can anticipate before help arrives. PMID- 16373122 TI - Air ambulance crews evacuate hundreds. PMID- 16373123 TI - EMS response on the Mississippi Coast. PMID- 16373124 TI - Mississippi welcomes an assist. PMID- 16373125 TI - Houston comes through for evacuees. PMID- 16373127 TI - Cric strikes a nerve. PMID- 16373128 TI - Conflict can be a good thing. PMID- 16373129 TI - STOP! I see dead people. PMID- 16373130 TI - Fried rice syndrome. PMID- 16373132 TI - Cardiac care on the street: how today's street medicine evolved from the Columbus Heartmobile & other pioneering projects. PMID- 16373135 TI - Opening the black box of post-stroke rehabilitation: stroke rehabilitation patients, processes, and outcomes. AB - DeJong G, Horn SD, Conroy B, Nichols D, Healton EB. Opening the black box of post stroke rehabilitation: stroke rehabilitation patients, processes, and outcomes. This article introduces the journal's supplement devoted to the methods and findings of the 7-site Post-Stroke Rehabilitation Outcomes Project (PSROP), a study designed to provide a very granular in-depth understanding of stroke rehabilitation practice and how practice is related to outcomes. The article summarizes current knowledge about the effectiveness of post-stroke rehabilitation, outlines where the PSROP fits into the broader traditions of stroke rehabilitation outcomes research, underscores the study's methodologic innovations, and summarizes the scope of the articles that follow. PMID- 16373136 TI - Another look at observational studies in rehabilitation research: going beyond the holy grail of the randomized controlled trial. AB - Horn SD, DeJong G, Ryser DK, Veazie PJ, Teraoka J. Another look at observational studies in rehabilitation research: going beyond the holy grail of the randomized controlled trial. This commentary compares randomized controlled trials (RCTs) and clinical practice improvement (CPI) approaches to study design, evaluates their relative advantages and disadvantages, and discusses their implications for rehabilitation research and evidence-based practice. Many argue that observational cohort studies are not sufficient as scientific evidence for practice change. We challenge this assertion by introducing the concept of a CPI study: a comprehensive observational paradigm structured to decrease biases generally associated with observational research. One strength of CPI studies is their attention to defining and characterizing the "black box" of clinical practice. CPI studies require demanding data collection, but by using bivariate and multivariate associations among patient characteristics, process steps, and outcomes, they can uncover best practices more quickly while achieving many of the presumed advantages of RCTs. PMID- 16373137 TI - Applying the clinical practice improvement approach to stroke rehabilitation: methods used and baseline results. AB - Gassaway J, Horn SD, DeJong G, Smout RJ, Clark C, James R. Applying the clinical practice improvement approach to stroke rehabilitation: methods used and baseline results. OBJECTIVES: To describe the methods used and baseline data for the Post Stroke Rehabilitation Outcomes Project (PSROP). DESIGN: Prospective observational cohort study. SETTING: Seven inpatient rehabilitation facilities (IRFs) in the United States and New Zealand. PARTICIPANTS: Consecutive convenience sample of 1291 poststroke rehabilitation patients, age older than 18, who were treated between 2001 and 2003 in 7 IRFs (1161 patients in 6 U.S. IRFs). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Change in FIM score, change in severity of illness, and discharge destination. RESULTS: For the U.S. sample, the average age was 66 years, 52% were men, 60% were white, and 23% were black. Medicare was the most frequent payer. Seventy-seven percent of strokes were ischemic, with 43% in the left brain, 44% in the right brain, and 11% bilateral. Mean admission total FIM score was 61, with a mean motor FIM score of 40 and mean cognitive FIM score of 21. Lower FIM scores are associated with higher severity-of-illness scores. Mean rehabilitation length of stay was 18.6 days; 78% of patients were discharged home. At discharge, the average increase in total FIM score was 26, in motor FIM score was 22, and in cognitive FIM score was 4. CONCLUSIONS: This article outlines methods used in the PSROP, provides an overview of participating IRFs, describes the database, and summarizes key characteristics to enable readers of subsequent articles to better interpret study findings and determine generalizability. PMID- 16373138 TI - Timing of initiation of rehabilitation after stroke. AB - Maulden SA, Gassaway J, Horn SD, Smout RJ, DeJong G. Timing of initiation of rehabilitation after stroke. OBJECTIVE: To study associations between days from stroke symptom onset to rehabilitation admission and rehabilitation outcomes, controlling for a variety of confounding variables. DESIGN: Observational cohort study of 200 consecutive post-stroke rehabilitation patients in each of 6 inpatient rehabilitation facilities. SETTING: Six U.S. inpatient rehabilitation hospitals. PARTICIPANTS: Patients (N=969) with moderate or severe strokes who had days from stroke symptom onset to rehabilitation admission recorded in their medical records. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Discharge total FIM, discharge motor FIM, discharge activities of daily living (ADL) FIM, and discharge mobility FIM scores, as well as rehabilitation length of stay (LOS). RESULTS: Fewer days from stroke symptom onset to rehabilitation admission was associated significantly with better functional outcomes: higher total, motor, mobility, and ADL discharge FIM scores, controlling for confounding variables. For severely impaired patients with stroke in case-mix groups (CMGs) 108-114, the relation was strongest, with F statistics greater than 24.1 for each functional outcome. For patients with moderately severe stroke in CMGs 104-107, fewer days from stroke symptom onset to rehabilitation admission was associated significantly with shorter rehabilitation LOS. CONCLUSIONS: Fewer days from stroke symptom onset to rehabilitation admission is associated with better functional outcomes at discharge and shorter LOS. PMID- 16373139 TI - Physical therapy during stroke rehabilitation for people with different walking abilities. AB - Latham NK, Jette DU, Slavin M, Richards LG, Procino A, Smout RJ, Horn SD. Physical therapy during stroke rehabilitation for people with different walking abilities. OBJECTIVE: To describe how physical therapy (PT) activities during post-stroke inpatient rehabilitation vary by admission walking ability and over time. DESIGN: Observational cohort study. SETTING: Six inpatient rehabilitation hospitals in the United States. PARTICIPANTS: People receiving post-stroke PT (N=715) who were classified as walking at admission. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Percentage of time spent in 11 activities, percentage of patients who participated in each activity, and the FIM instrument scores. RESULTS: The majority of PT time was spent in gait activities. Even people with the most limited mobility spent 25% to 38% of PT time in gait activities during the first 6-hour treatment block. Treatment progression was evident, and a shift to more advanced activities occurred over time (eg, less bed mobility and more advanced gait). However, even in the final 6-hour block, a small proportion of time was spent on community mobility activities (1.2%-5.2%), and most people received no community mobility training. CONCLUSIONS: PT activities focused on specific functional tasks at the ability level of each individual patient and provided higher-level activities as patients improved their function. However, although there is increasing recognition that the environment influences task performance, little time was spent in community mobility activities before discharge. PMID- 16373140 TI - Characterizing occupational therapy practice in stroke rehabilitation. AB - Richards LG, Latham NK, Jette DU, Rosenberg L, Smout RJ, DeJong G. Characterizing occupational therapy practice in stroke rehabilitation. OBJECTIVES: To describe how occupational therapy (OT) activities during stroke inpatient rehabilitation vary by admission functional status and over time and how time spent in these various activities relates to functional status at discharge. DESIGN: Observational cohort study. SETTING: Six inpatient rehabilitation hospitals in the United States. PARTICIPANTS: People (N=713) receiving 4 to 19 hours of post stroke OT. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Patients were categorized by number of 4-hour blocks of OT received and by admission upper extremity (UE) dressing score on the FIM instrument. In each group, the percentage of time spent in 16 activities and the percentage of patients who received each activity were calculated. The amount of time in activities was compared for those patients scoring 1 or 2 at admission who achieved at least a level of supervision for UE dressing (a score of >or=5) using Wilcoxon 2-sample tests. RESULTS: The majority of OT time was spent in impairment-focused activities (37.5%) or training basic activities of daily living (31.9%). Treatment progressed to more advanced activities over time (eg, less bed mobility, more home management), yet little time was spent on community integration or leisure activities and with very few patients. Successful patients received more higher-level activities, whereas unsuccessful patients received larger amounts of basic-level activities. CONCLUSIONS: OT activities focused on a combination of remediating impairments and retraining specific functional tasks, at the ability level of each individual patient, and provided higher-level activities as patients improved their function. More time in higher-level activities was related to greater success in rehabilitation. However, higher level activities remain the least common activities provided during inpatient rehabilitation. PMID- 16373141 TI - Characterizing speech and language pathology outcomes in stroke rehabilitation. AB - Hatfield B, Millet D, Coles J, Gassaway J, Conroy B, Smout RJ. Characterizing speech and language pathology outcomes in stroke rehabilitation. OBJECTIVES: To describe a subset of speech-language pathology (SLP) patients in the Post-Stroke Rehabilitation Outcomes Project and to examine outcomes for patients with low admission FIM levels of auditory comprehension and verbal expression. DESIGN: Observational cohort study. SETTING: Five inpatient rehabilitation hospitals. PARTICIPANTS: Patients (N=397) receiving post-stroke SLP with admission FIM cognitive components at levels 1 through 5. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Increase in comprehension and expression FIM scores from admission to discharge. RESULTS: Cognitively and linguistically complex SLP activities (problem-solving and executive functioning skills) were associated with greater likelihood of success in low- to mid-level functioning communicators in the acute post-stroke rehabilitation period. CONCLUSIONS: The results challenge common clinical practice by suggesting that use of high-level cognitively and linguistically complex SLP activities early in a patient's stay may result in more efficient practice and better outcomes regardless of the patient's functional communication severity level on admission. PMID- 16373142 TI - An exploration of central nervous system medication use and outcomes in stroke rehabilitation. AB - Conroy B, Zorowitz R, Horn SD, Ryser DK, Teraoka J, Smout RJ. An exploration of central nervous system medication use and outcomes in stroke rehabilitation. OBJECTIVE: To study associations between neurobehavioral impairments, use of neurotropic medications, and outcomes for inpatient stroke rehabilitation, controlling for a variety of confounding variables. DESIGN: Observational cohort study of post-stroke rehabilitation. SETTING: Six inpatient rehabilitation hospitals in the United States. PARTICIPANTS: Patients with moderate or severe strokes (N=919). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Discharge disposition, FIM score change, and rehabilitation length of stay (LOS). RESULTS: Neurobehavioral impairments and use of many medications, including first generation selective serotonin reuptake inhibitors, older traditional antipsychotic medications, and anti-Parkinsonian neuro-stimulants, have a statistical association with poorer outcomes, whereas use of the atypical antipsychotic medications has a positive association with improvement in motor FIM scores. Counter-intuitively, use of opioid analgesics is associated with a larger motor FIM score change but not an increase in LOS or reduced percentage of discharge to community. There was significant variation in use of neurotropic medications among the 6 study sites during inpatient stroke rehabilitation. CONCLUSIONS: There are many opportunities to enhance a stroke survivor's ability to benefit from acute inpatient stroke rehabilitation through improved understanding of associations of neurotropic medications with outcomes for different patient groups. PMID- 16373143 TI - Nutrition support (tube feeding) as a rehabilitation intervention. AB - James R, Gines D, Menlove A, Horn SD, Gassaway J, Smout RJ. Nutrition support (tube feeding) as a rehabilitation intervention. OBJECTIVE: To describe site variation in use of enteral feeding and its association with stroke rehabilitation outcomes, controlling for a variety of confounding variables. DESIGN: Prospective observational cohort study. SETTING: Six inpatient rehabilitation facilities in the United States. PARTICIPANTS: Patients (N=919) from the Post-Stroke Rehabilitation Outcomes Project database with moderate or severe stroke who were discharged to home, community, or skilled nursing facility. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Change in total, motor, and cognitive FIM instrument scores and change in severity of illness. RESULTS: Monitoring of nutritional status and the frequency of tube-feeding interventions for patients with moderate and severe stroke varied significantly among sites. Patients with tube feeding had higher severity of illness and lower functioning on admission compared with patients who did not receive tube feeding. However, when we controlled for severity of illness, admission FIM score, and other important covariates, we found that patients with severe strokes who were tube fed for more than 25% of their stay had greater increases in total, motor, and cognitive FIM scores and greater improvement in severity of illness by discharge. CONCLUSIONS: Nutrition support (tube feeding) is an effective therapy in rehabilitation service for patients with severe strokes and is associated with greater motor and cognitive improvements, even in patients with the most severe strokes. PMID- 16373144 TI - The early impact of the inpatient rehabilitation facility prospective payment system on stroke rehabilitation case mix, practice patterns, and outcomes. AB - DeJong G, Horn SD, Smout RJ, Ryser DK. The early impact of the inpatient rehabilitation facility prospective payment system on stroke rehabilitation case mix, practice patterns, and outcomes. OBJECTIVE: To determine the early effects of the inpatient rehabilitation facility (IRF) prospective payment system (PPS) on stroke rehabilitation case mix, practice patterns, and outcomes. DESIGN: Prospective observational cohort study. SETTING: Three IRFs in the United States. PARTICIPANTS: Consecutively enrolled convenience sample of 539 stroke rehabilitation patients treated between 2001 and 2003 in 3 IRFs. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Length of stay (LOS), therapy utilization, FIM instrument gain, and discharge destination. RESULTS: The IRF-PPS had no material short-term effect on stroke rehabilitation case mix and LOS for the study facilities. Facilities shifted physical and occupational therapy resources from those in the most severe case-mix groups (CMGs) to those in the moderate CMGs. Those in the more severe CMGs also experienced a noticeable decline in FIM score gain over the course of the rehabilitation stay. Using multivariate analyses, the authors discerned no major role for the IRF-PPS in explaining pre- and post-PPS differences in utilization and outcome among study facilities. CONCLUSIONS: For the 3 study facilities, IRF-PPS did not materially reshape stroke rehabilitation case mix, utilization, and outcome in the early stages of PPS implementation, apart from the shift in therapy resources from more severely involved stroke patients to moderately involved patients. The study's findings are limited to 3 facilities, and a longer time horizon is needed to more fully determine the effects of the IRF-PPS. PMID- 16373145 TI - Stroke rehabilitation patients, practice, and outcomes: is earlier and more aggressive therapy better? AB - Horn SD, DeJong G, Smout RJ, Gassaway J, James R, Conroy B. Stroke rehabilitation patients, practice, and outcomes: is earlier and more aggressive therapy better? OBJECTIVE: To examine associations of patient characteristics, rehabilitation therapies, neurotropic medications, nutritional support, and timing of initiation of rehabilitation with functional outcomes and discharge destination for inpatient stroke rehabilitation patients. DESIGN: Prospective observational cohort study. SETTING: Five U.S. inpatient rehabilitation facilities. PARTICIPANTS: Post-stroke rehabilitation patients (N=830; age, >18 y) with moderate or severe strokes, from the Post-Stroke Rehabilitation Outcomes Project database. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Discharge total, motor, and cognitive FIM scores and discharge destination. RESULTS: Controlling for patient differences, various activities and interventions were associated with better outcomes including earlier initiation of rehabilitation, more time spent per day in higher-level rehabilitation activities such as gait, upper extremity control, and problem solving, use of newer psychiatric medications, and enteral feeding. Several findings part with conventional practice, such as starting gait training in the first 3 hours of physical therapy, even for low level patients, was associated with better outcomes. CONCLUSIONS: Specific therapy activities and interventions are associated with better outcomes. Earlier rehabilitation admission, higher-level activities early in the rehabilitation process, tube feeding, and newer medications are associated with better stroke rehabilitation outcomes. PMID- 16373146 TI - A comparison of stroke rehabilitation practice and outcomes between New Zealand and United States facilities. AB - McNaughton H, DeJong G, Smout RJ, Melvin JL, Brandstater M. A comparison of stroke rehabilitation practice and outcomes between New Zealand and United States facilities. OBJECTIVE: To compare stroke rehabilitation practice and outcomes between New Zealand (NZ) and the United States. DESIGN: Prospective observational cohort study. SETTING: Seven inpatient rehabilitation facilities (IRFs) in the United States and NZ. PARTICIPANTS: Consecutive convenience sample of 1161 patients in 6 U.S. IRFs and 130 in 1 NZ IRF (age, >18 y) after acute stroke. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Change in FIM score and discharge destination. RESULTS: NZ participants were older than U.S. participants (mean: 74.1 y vs 66.0 y, respectively; P<.001). Measures of initial stroke severity were higher for U.S. participants. Mean rehabilitation length of stay (LOS) was shorter for U.S. participants (18.6d vs 30.0 d, P<.001), but physical and occupational therapy time per patient was considerably higher despite the shorter LOS. U.S. therapists were involved in more active therapies for more of the time. Outcomes were better for U.S. participants, with fewer discharged to institutional care (13.2% vs 21.5%, P=.006) and larger changes in FIM scores. CONCLUSIONS: U.S. participants with acute stroke who were selected for rehabilitation had better outcomes than NZ participants, despite shorter stays in the rehabilitation facility. U.S. participants had more intensive input from physiotherapists and occupational therapists, which may explain some of the larger increases in FIM scores. This suggests that further studies with tighter controls on case mix may add additional information on the effects of therapy intensity on patients with stroke. PMID- 16373147 TI - The post-stroke rehabilitation outcomes project. AB - Ottenbacher KJ. The Post-Stroke Rehabilitation Outcomes Project. The Post-Stroke Rehabilitation Outcomes Project (PSROP) examined a large sample of patients from multiple facilities receiving inpatient stroke rehabilitation services. This commentary describes strengths and potential limitations of the investigation including selection bias, observation bias, confounds, and interpretation. The PSROP is an important study that will advance our understanding of effective treatment for persons with stroke. PMID- 16373148 TI - The post-stroke rehabilitation outcomes project. AB - Jette AM. The Post-Stroke Rehabilitation Outcomes Project. The Post-Stroke Rehabilitation Outcomes Project (PSROP) provides an important example of the value of observational study designs in rehabilitation. The strength of the PSROP lies in the extensive, in-depth data collected on the specific rehabilitation interventions provided to patients and their relationship to short-term outcomes as well as the wide generalizability of the study's findings. Although providing valuable insights, one has to be extremely cautious in drawing direct practice recommendations from the PSROP given several internal validity threats inherent in the PSROP design. PMID- 16373151 TI - Simulation training for surgical residents: cool computers or core curriculum? PMID- 16373152 TI - PET scans in tumor staging: hope or hype? PMID- 16373153 TI - Cowden syndrome: report of two cases and review of clinical presentation and management of a rare colorectal polyposis. AB - BACKGROUND/AIMS: Cowden syndrome (CS) is a rare and complex disease inherited through an autosomal dominant trait associated with germline mutations of the PTEN gene. OBJECTIVE: This article reports 2 female patients with classic features of the syndrome and reviews the current guidelines regarding diagnosis and surveillance. REVIEW: Although it exhibits variable clinical expressivity, the diagnosis is based on characteristic mucocutaneous alterations such as multiple facial trichilemmomas, oral mucosal papillomatosis, and acral and palmoplantar keratoses. These manifestations often precede systemic involvement. Extracutaneous lesions include fibrocystic disease of the breast, thyroid goiters or adenomas, multiple polyposis of the gastrointestinal tract, and ovarian cysts. Gastrointestinal polyps are usually asymptomatic, and the risk of gastrointestinal cancer is not greatly increased. Otherwise, an important feature of Cowden's disease is the greater risk of breast and thyroid cancer. CONCLUSIONS: Because of the potentially serious associations with internal malignancy, early and accurate diagnosis of CS is essential. For this reason, all patients must be screened for occult malignancies and undergo close surveillance throughout lifetime. PMID- 16373154 TI - The management of bile duct injuries occurring during laparoscopic cholecystectomy. PMID- 16373155 TI - Lateral neck mass in a young woman. PMID- 16373156 TI - Jerome P. Webster, MD (1888-1974). PMID- 16373157 TI - Continuity of care in the 80-hour workweek era? PMID- 16373158 TI - Intraabdominal pulmonary sequestration. AB - Bronchopulmonary sequestration is an uncommon developmental abnormality that frequently presents as an incidental mass. Differential diagnosis includes malignancy. Reports of extralobar abdominal pulmonary sequestration in adults are sparse. Only 2.5% of all pulmonary sequestrations are detected below the diaphragm. The authors present a case of a 45-year-old woman and a literature review. Computed tomography, magnetic resonance imaging, and operative and pathologic findings are provided. A review of diagnosis and therapy is presented. PMID- 16373159 TI - Partial thyroidectomy: illustrated reflections for surgical residents. PMID- 16373160 TI - Radiofrequency ablation and plication--a new technique for prolapsing hemorrhoidal disease. AB - BACKGROUND: The author describes a modified procedure of ablation with a radiofrequency device and plication of the hemorrhoidal mass for prolapsing hemorrhoids. The study is aimed at ascertaining if this procedure provides any advantages over the conventional hemorrhoid surgery. MATERIALS AND METHODS: Two different studies are included. The first study describes 600 serial patients with prolapsing hemorrhoids treated with this technique over a period of 18 months. An Ellman radiofrequency generator was used for the ablation of the hemorrhoids. The operative technique and postoperative outcome is reported. The second study compares this technique with standard Milligan-Morgan hemorrhoidectomy in a randomized trial of 100 patients. RESULTS: With this new procedure, the post-defecation pain and pain at rest were within tolerable limits (pain scores 1 to 4 on visual analog scale). Post-defecation bleeding was present in 60% of the patients. Pruritus and perianal thrombosis were complained by few others. No patient encountered any incontinence, prolapse, or stenosis. The comparative study showed definite advantages of this modified technique over Milligan-Morgan hemorrhoidectomy. CONCLUSION: The procedure of radiofrequency ablation and plication of hemorrhoids restricts the hospital stay to only a few hours and provides rapid physical recovery. It does seem to be a better alternative to the conventional surgical procedures in terms of postoperative pain, return to work, and complications. PMID- 16373161 TI - Surgical experience and cosmetic outcomes in children with sacrococcygeal teratoma. AB - To determine the extent of progress in the treatment of sacrococcygeal teratomas (SCTs), the experience of 25 patients over a period of 30 years between 1972 and 2002 was evaluated. The medical records of the children were reviewed for demographics, histology, clinical course, bladder, anorectal and sexual function, and outcome. In particular, analysis of long-term cosmetic results after surgery for SCT was performed. The follow-up period ranged from 1 day to 396 months (median, 212 months). In more than half of the cases, unsatisfactory cosmetic long-term results after surgery for SCT were found. This study demonstrates that poor cosmetic results in the buttock region were the most common long-term complication after surgery for SCT, possibly leading to further corrective operations in later life. A better surgical procedure for closing the buttock region in the treatment of SCT should be discussed. Early assessment of bladder, anorectal, and sexual function along with long-term cosmetic results should be integrated into the well-established long-term oncology follow-up program. Moreover one has to keep in mind that cesarean section could become necessary as the primary delivery technique in female patients who have had surgery for SCT. More evaluations concerning this aspect are necessary. PMID- 16373163 TI - Medical malpractice matters: organization of the medical record notes. PMID- 16373162 TI - The why and how of coordinator certification. PMID- 16373164 TI - My surgical mentor. PMID- 16373165 TI - My surgery mentor... a tapestry. PMID- 16373166 TI - My mentors. PMID- 16373167 TI - Impact of resident duty-hour reform on faculty clinical productivity. AB - OBJECTIVE: Prior data have shown that resident duty-hour reform has not affected faculty work hours; yet the preservation of faculty hours may have been at the expense of productivity. We sought to examine change in clinical productivity. DESIGN: Anonymous survey and analysis of faculty relative value units (RVU) database. SETTING: A single, large academic medical center. PARTICIPANTS: All clinical faculty in the Department of Surgery. METHODS: An anonymous survey was distributed to surgical faculty 18 months after reform and compared with surveys taken before and after reform. Opinions regarding productivity and working hours were solicited. P values were determined by chi-square or Student t-tests. Relative value unit data, reflecting clinical productivity, were compared before and after reform. Regression was performed with dependent variable "lnRVU" and independent variables "calendar month," "pre/post" July 2003, and "surgeon." The coefficient on "pre/post" reflected average change in RVUs. RESULTS: A total of 49 of 73 surveys were returned (67% response). Faculty reported an average of 68.0+/-7.0 weekly work hours (p=NS compared with previous survey). In the current survey, 35% felt their overall productivity had fallen due to reform. Among these, 83% felt academic productivity had suffered, 11% were unsure, and 1 person (6%) believed academic productivity was preserved. The majority (82%) reported preserved clinical productivity, 6% reported a decrease, and 12% were unsure. Overall, 60% reported doing work previously done by residents. When RVU data were examined, the coefficient on change pre- and post-reform indicated a 5.7% increase in productivity (p=0.005). However, this effect was driven by 5 surgeons with a greater than 75% increase in productivity, all young faculty, early in practice. Excluding these, there was no significant change (0.6% increase, p=0.77). CONCLUSIONS: Faculty have preserved work hours and clinical productivity, despite a tendency to take on work previously done by residents. This suggests that academic activities may have suffered. PMID- 16373168 TI - Ongoing faculty evaluations: developmental gain or just more pain? AB - PURPOSE: We continue to increase the amount of evaluations to improve the outcomes of our residency programs. Although ongoing faculty evaluations clearly are an important part of faculty development, their value in terms of improving the program needs to be evaluated. The questions asked were as follows: (1) Do faculty evaluations continue to improve the faculty over the course of successive evaluation periods? (2) Are there groups of faculty who would benefit the most from faculty evaluation feedback? (3) Are there any specific objective categories within the evaluation that carry more value and may help to shorten this form? METHODS: Forty-two faculty members were evaluated by 40 surgical residents with an assessment form developed by surgical residents that assessed faculty members by 10 different criteria. The initial set of data was collected, and attending surgeons were given an intervention in the form of a letter detailing how they had been assessed in each of the 10 categories. The attending surgeons were evaluated again 6 months later and were given an intervention in the form of verbal feedback regarding their evaluations. The attending faculty members were then assessed 1 year after that. One way analyses of variance and Fisher Protected Least Significant Difference (PLSD) were used to analyze the resulting data to determine if there were significant differences in the faculty evaluations. A part-whole correlation was performed that correlated the 10 evaluation criteria against the mean score on each evaluation, and partial eta squared analysis was used to determine which criterion had the largest effect on the overall means. RESULTS: The means for the 42 faculty members as a whole continued to improve from the first to the final evaluation period, with 30 faculty members increasing their mean score (18 significantly) and 12 decreasing their mean score (4 significantly). Seven of the 10 evaluation criteria's means improved sequentially by feedback session. These were (1) Didactic Teaching, (2) Teaching Rounds, (3) Attendance at Didactic Activities, (4) Allows [Resident] Autonomy to Make Independent Decisions, (5) Provides Feedback, (6) Stimulates Critical Thinking with Use of Literature, and (7) Encourages and Maintains an Atmosphere of Professional Mutual Respect for All Members of Health Care Team (Role Model). The faculty group with the lowest evaluations improved significantly more than those of both the middle and the role model group, with the middle and the role model groups improving, but not significantly differently from each other. Of the criteria that improved, only Provides Feedback improved significantly from the first to second and the second to third evaluation periods. The three criteria with the highest correlation coefficients were Role Model (0.76), Provides Feedback (0.75), and Stimulates Critical Thinking (0.74). The results from the partial eta-squared test showed that the criterion with the largest effect size was Provides Feedback (0.28). These analyses indicate that the criterion Provides Feedback was both highly correlated with the average score on a faculty member's evaluation and was more responsible than any other criteria for the overall improvement in the mean evaluation score of the faculty members. CONCLUSIONS: (1) Ongoing faculty evaluations indeed are a powerful tool to improve the faculty as a whole. (2) The faculty members with the lowest evaluations showed the largest amount of improvement. (3) Providing feedback to the residents seems to be the most valued factor by the residents for faculty evaluations and perhaps could become the basis of the evaluation for the most accomplished faculty. PMID- 16373169 TI - Preliminary general surgery residents: indentured servitude or golden opportunity? AB - BACKGROUND: One-year, preliminary general surgery (GS) positions are viewed by medical student applicants as among the least desirable positions offered through the match. Hundreds of preliminary GS positions go unfilled each year, and international medical graduates (IMGs) fill a substantial portion of the spots filled. "Prelims" foster diversity and add manpower to surgical training programs, but do trainees eventually benefit from this 1-year effort? METHODS: Nondesignated (ND) preliminary GS residents at the Mayo Clinic, Rochester, Minnesota, were identified from 1993 to 2003. Trainees were followed (telephone, e-mail, Internet registries) looking at subsequent training, specialty choice, and eventual practice location. RESULTS: From 118 ND preliminary GS residents, the current data of 105 (46 U.S. medical graduates, 59 IMGs) trainees are known. Thirteen trainees (11%) progressed on to a second year of training (location: United States=10, international=3), but they could not be located thereafter. Two additional prelims chose another profession. Seventy one of the remaining 103 prelims (70%) advanced to other non-Mayo residency programs to continue or finish their training in general surgery (n=22), a surgical subspecialty (n=26), or some other field (n=23). Thirty-two prelims remained at Mayo Clinic-Rochester in GS (n=16), a surgical subspecialty (n=12), or a nonsurgical field (n=4). From the 59 IMGs, 52 remained in the United States, whereas 7 left for another country to practice medicine. CONCLUSION: Most of our ND preliminary GS residents progress toward completion of surgical or medicine residencies. Although initially described as a finite period of training, the 1-year preliminary GS residency offers both U.S. and international students opportunity for career advancement. PMID- 16373170 TI - Mutations in vimentin disrupt the cytoskeleton in fibroblasts and delay execution of apoptosis. AB - To get new insights into the function of the intermediate filament (IF) protein vimentin in cell physiology, we generated two mutant cDNAs, one with a point mutation in the consensus motif in coil1A (R113C) and one with the complete deletion of coil 2B of the rod domain. In keratins and glia filament protein (GFAP), analogous mutations cause keratinopathies and Alexander disease, respectively. Both mutants prevented filament assembly in vitro and inhibited assembly of wild-type vimentin when present in equal amounts. In stably transfected preadipocytes, these mutants caused the complete disruption of the endogenous vimentin network, demonstrating their dominant-negative behaviour. Cytoplasmic vimentin aggregates colocalised with the chaperones alphaB-crystallin and HSP40. Moreover, vimR113C mutant cells were more resistant against staurosporine-induced apoptosis compared to controls. We hypothesise that mutations in the vimentin gene, like in most classes of IF genes, may contribute to distinct human diseases. PMID- 16373171 TI - Transdifferentiation of mesenchymal stem cells into Schwann cell-like myelinating cells. AB - Bone marrow stromal cells (MSC) are multipotent stem cells that differentiate into cells of the mesodermal lineage. Although adult, their differentiation potential is remarkable, and they are able to transdifferentiate. Transdifferentiated cultivated rat MSC (tMSC) changed morphologically into cells resembling typical spindle-shaped Schwann cells (SC) with enhanced expression of LNGF receptor, Krox-20, CD104 and S100beta protein and decreased expression of bone morphogenetic protein receptor-1A compared to untreated rat MSC (rMSC). Transdifferentiation was reversible and repeatable. To evaluate the myelinating capacity, rMSC, tMSC, or SC cultured from male rats were grafted into an autologous muscle conduit bridging a 2-cm gap in the female rat sciatic nerve. The presence of the male-specific SRY gene (as revealed by PCR analysis) and S100 immunoreactivity of pre-labeled tMSC confirmed the presence of the implanted cells in the grafts. Three weeks after grafting, an appropriate regeneration was noted in the SC and in the tMSC groups, while regeneration in the rMSC group and in the control group without any cells was impaired. In contrast to SC, in some cases, single tMSC were able to myelinate more than one axon. Our findings demonstrate that it may be possible to differentiate MSC into therapeutically useful cells for clinical applications. PMID- 16373172 TI - Post-translational amino acid racemization in the frog skin peptide deltorphin I in the secretion granules of cutaneous serous glands. AB - The dermal glands of the South American hylid frog Phyllomedusa bicolor synthesize and expel huge amounts of cationic, alpha-helical, 24- to 33-residue antimicrobial peptides, the dermaseptins B. These glands also produce a wide array of peptides that are similar to mammalian hormones and neuropeptides, including a heptapeptide opioid containing a D-amino acid, deltorphin I (Tyr-DAla Phe-Asp-Val-Val-Gly NH2). Its biological activity is due to the racemization of L Ala2 to D-Ala. The dermaseptins B and deltorphins are all derived from a single family of precursor polypeptides that have an N-terminal preprosequence that is remarkably well conserved, although the progenitor sequences giving rise to mature opioid or antimicrobial peptides are markedly different. Monoclonal and polyclonal antibodies were used to examine the cellular and ultrastructural distributions of deltorphin I and dermaseptin B in the serous glands by immunofluoresence confocal microscopy and immunogold-electron microscopy. Preprodeltorphin I and preprodermaseptins B are sorted into the regulated pathway of secretion, where they are processed to give the mature products. Deltorphin I, [l-Ala2]-deltorphin I and dermaseptin B are all stored together in secretion granules which accumulate in the cytoplasm of all serous glands. We conclude that the L- to D-amino acid isomerization of the deltorphin I occurs in the secretory granules as a post-translational event. Thus the specificity of isomerization depends on the presence of structural and/or conformational determinants in the peptide N-terminus surrounding the isomerization site. PMID- 16373173 TI - Characterization of the endogenous GIT1-betaPIX complex, and identification of its association to membranes. AB - G protein-coupled receptor kinase interactors (GITs) are adaptor proteins with ADP-ribosylating factor--GTPase-activating protein (ARF-GAP) activity that form complexes with the p21-activated kinase-interacting exchange factor (PIX) guanine nucleotide exchanging factors for Rac and Cdc42. In this study we have characterized the endogenous GIT1/p95-APP1/Cat1 (GIT1)- PIX complexes in neuronal and non-neuronal cells. In COS7 cells, immunocytochemical analysis shows the localization of endogenous GIT1 in the perinuclear region of the cell, as well as at the cell periphery, where GIT1 co-localizes with filamentous actin. The perinuclear localization of endogenous GIT1 was confirmed in avian fibroblasts. In COS7 cells, immunoprecipitation and microsequencing experiments with either anti-GIT1 or anti-betaPIX antibodies unequivocally show that betaPIX is uniquely associated with GIT1 in lysates from these cells, while GIT2/PKL/p95-APP2/Cat2 (GIT2) is undetectable in the endogenous complexes. Moreover, this analysis demonstrates that betaPIX is the limiting factor for the formation of the endogenous complexes, since a small fraction of GIT1 can be co-immunoprecipitated with most betaPIX from these cells. Saponin treatment of unfixed cells indicates that betaPIX-bound GIT1 is preferentially retained in the saponin-resistant fraction when compared to betaPIX-free GIT1. Moreover, analysis by tissue fractionation shows that a significant fraction of the endogenous GIT1-betaPIX complex is firmly associated to membranes from brain homogenates. Our findings show the specific localization of the complex at intracellular membranes, and indicate a correlation between the association of GIT1 to betaPIX, and the localization of the endogenous complex at membranes. PMID- 16373177 TI - Physical activity--the "new" nutrition guideline. PMID- 16373174 TI - Characterization of the oligosaccharide component of alpha3beta1 integrin from human bladder carcinoma cell line T24 and its role in adhesion and migration. AB - Malignant transformation is highly associated with altered expression of cell surface N-linked oligosaccharides. These changes concern integrins, a family of cell surface glycoproteins involved in the attachment and migration of cells on various extracellular matrix proteins. The integrin alpha3beta1 is particularly interesting because of its role in migration and invasion of several types of metastatic tumours. In this study, alpha3beta1 from human bladder T24 carcinoma cells was purified and treated with peptide N-glycosidase F. Then the N-glycans of the alpha3 and beta1 subunits were characterized using matrix-assisted laser desorption ionization mass spectrometry (MALDI MS). In alpha3beta1 integrin the presence of high-mannose, hybrid and predominantly complex type N oligosaccharides was shown. Unlike to normal epithelium cells, in both subunits of alpha3beta1 integrin from cancer cells, the sialylated tetraantennary complex type glycan Hex7HexNAc6FucSia4 was present. In a direct ligand binding assay, desialylated alpha3beta1 integrin exhibited significantly higher fibronectin binding capability than untreated integrin, providing evidence that sialic acids play a direct role in ligand-receptor interaction. Moreover, alpha3beta1 integrin was shown to take part in T24 cell migration on fibronectin: anti-alpha3 antibodies induced ca 30% inhibition of wound closure. Treatment of T24 cells with swainsonine reduced the rate of bladder carcinoma cell migration by 16%, indicating the role of beta1,6 branched complex type glycans in this process. Our data show that alpha3beta1 integrin function may be altered by glycosylation, that both subunits contribute to these changes, and that glycosylation may be considered a newly found mechanism in the regulation of integrin function. PMID- 16373178 TI - Trials and tribulations: a small pilot telehealth home care program for medicare patients. AB - This article describes a home care agency's experience initiating the technology of a telehealth program for a selected view of its home care patients. The goal of the telehealth program was to improve patient outcomes by augmenting patients' regularly scheduled in-home skilled nursing visits with video-conferencing encounters. Patient selection, costs, projected savings, patient satisfaction, and the technical, clinical, and patient problems with the telehealth system are discussed. PMID- 16373180 TI - Challenges of doing intervention research with the elderly taiwanese population: example of a tai chi/movement therapy. AB - The purpose of this article is to address the challenges of conducting intervention research with an elderly population by using the example of a tai chi intervention study. Various difficulties can arise while conducting interventions with the elderly; these can involve factors related to long-term care facilities, health care providers, and the research team, as well as the institutionalized elders themselves. Any difficulty may influence research results. As a consequence, challenges must be dealt with in a positive manner to conduct a valid geriatric study. Modifications and strategies that the research team engaged to manage the challenges in this study are discussed. PMID- 16373181 TI - Health and social care policy for the elderly in Belgium. AB - Current Belgian health care policy for the elderly can be classified broadly into institutional care, community care, and home care. Economic restrictions, smaller families, increased employment among women, geographic distances between family members, and the preference of the elderly to remain at home are factors challenging Belgian policy makers and health professionals to restructure the current health care system and provide integrated holistic health and social care to the elderly. These challenges are being addressed through initiatives to customize care for the elderly at home, in the community, or in care environments by improving coordination of care, programs of support for geriatric care and dementia, and research into new accommodation and health solutions. PMID- 16373182 TI - Integrative review of research related to meditation, spirituality, and the elderly. AB - Recently, increased attention has been given to meditation-relaxation strategies to improve physical health, reduce pain, enhance immune response, improve emotional well-being, and foster spiritual growth. This article reviews research conducted in the last 25 years about meditation and spirituality, in particular as it relates to the health of the elderly. This review supports the hypothesis that meditation can be taught to the elderly, even those with dementia. The results also support the hypothesis that meditation and spiritual practices could promote significant social and emotional benefits for those in social isolation. Specific treatment plan interventions for nursing homes are discussed. Future research should investigate the effectiveness of various types of meditative and spiritual practices to nursing home residents. PMID- 16373183 TI - Effects of a comprehensive educational program on quality of life and emotional issues of dementia patient caregivers. AB - The aim of this community-based pre-post interventional study was to investigate the effectiveness of a comprehensive educational program reinforced by an individualized component (CEPRIC) on problems likely to be experienced by caregivers of persons with Alzheimer's disease, as defined by the North American Nursing Diagnosis Association (NANDA). The Beck Depression Scale, Beck Anxiety Inventory, and the Duke scales were used to measure depression, anxiety, and quality of life, respectively. Sixty-four participants (32 caregiver-patient pairs) took part in the program. This study suggests that caregiver problems (disturbed sleep and eating patterns, trauma risk, anxiety) were reduced and caregiver depression and anxiety scores were diminished; quality of life of caregivers was increased by the intervention. In conclusion, the CEPRIC is a viable option for Alzheimer's caregivers' education programs, particularly in an environment with limited respite care options. PMID- 16373184 TI - Cell membrane microparticles in blood and blood products: potentially pathogenic agents and diagnostic markers. AB - Cell membrane microparticles (MPs) circulate in the blood of healthy donors, and their elevated counts have been documented in various pathologies. Microparticles are phospholipid microvesicles of 0.05 to 1.5 microm in size, containing certain membrane proteins of their parental cells. Thus, different phenotypes of MPs derived from platelets, blood cells, endothelial cells, and some other cell types have been identified in plasma. Microparticles are released by various stimuli including shear stress, complement attack, or proapoptotic stimulation. Microparticle release is a highly controlled process and likely independent from metabolic energy. Elevated MPs in various diseases indicate their diagnostic importance, particularly in vascular pathologies. Moreover, MPs in blood possess a broad spectrum of biologic activities. Microparticles may facilitate cell-to cell interactions, induce cell signaling, or even transfer receptors between different cell types. The physiological roles of MPs in various tissue defense processes have been suggested and the pathophysiologic implications of MPs in thrombosis, inflammation, cancer metastasis, or response to pathogens have been proposed. This is important for transfusion medicine because MPs are present in both plasma and cellular blood products. Thus, the investigation of potentially pathogenic effects of MPs in blood products and of MP release associated with blood product processing and storage have yet to come. PMID- 16373185 TI - The role of erythropoietin therapy in the critically ill. AB - Critically ill patients receive an extraordinarily large number of blood transfusions. Between 40% and 50% of all patients admitted to intensive care units (ICUs) receive at least one allogeneic red blood cell (RBC) unit and average close to 5 U of RBCs during their ICU admission. RBC transfusion is not risk-free, and there is little evidence that "routine" transfusion of stored allogeneic RBCs is beneficial to critically ill patients. It is clear that most critically ill patients can tolerate hemoglobin levels as low as 7 g/dL, and therefore, a more conservative approach to RBC transfusion is warranted. Anemia of critical illness is a distinct clinical entity characterized by blunted erythropoietin (EPO) production and abnormalities in iron metabolism identical to what is commonly referred to as anemia of chronic disease. As such, the bone marrow in many of these patients responds to the administration of exogenous EPO, in spite of their underlying critical illness. The efficacy of perioperative recombinant human erythropoietin (rHuEPO) has been demonstrated in a variety of elective surgical settings. Similarly, in critically ill patients, rHuEPO therapy will also stimulate erythropoiesis. In randomized placebo-controlled trials, therapy with rHuEPO resulted in a significant reduction in allogeneic RBC transfusions. Strategies to increase the production of RBCs are complementary to other approaches to reduce blood loss in the ICU and decrease the transfusion threshold in the management of all critically ill patients. PMID- 16373186 TI - Controversies in the management of the anemia of prematurity using single-donor red blood cell transfusions and/or recombinant human erythropoietin. AB - Many controversial questions regarding the practice of neonatal red blood cell (RBC) transfusions exist, so that practices and policies vary widely. This article will critically assess information pertaining to two of these controversies, namely, the transfusion of RBCs stored for up to 42 days after collection vs the transfusion of fresh RBCs stored 7 days or less after donation and the use of recombinant human erythropoietin (rHuEPO) in attempts to either diminish the severity of or to treat the anemia of prematurity. Based on both theoretical considerations and several published clinical trials, RBCs from one donor stored up to 42 days in extended storage preservative solutions can safely provide all RBCs needed by most infants for small-volume transfusions. Based on a large number of clinical trials and a meta-analysis of these trials, it is impossible to provide firm guidelines for the use of rHuEPO in the treatment of the anemia of prematurity. Clearly, rHuEPO has efficacy in stimulating erythropoiesis in preterm infants, but success in the elimination or marked reduction in the need for RBC transfusions has not been definitively demonstrated. PMID- 16373187 TI - The role of photochemical treatment with amotosalen and UV-A light in the prevention of transfusion-transmitted cytomegalovirus infections. AB - Primary cytomegalovirus (CMV) infection is usually asymptomatic in immunocompetent patients but can cause serious life-threatening complications in immunocompromised CMV-seronegative patients, including patients receiving a bone marrow or peripheral blood stem cell transplant, recipients of some solid-organ transplants, and low-birth-weight neonates. Current recommendations for preventing transfusion-transmitted CMV (TT-CMV) infection in these patients include exclusive use of CMV-seronegative and/or leukoreduced cellular blood components (red blood cells and platelets) for transfusion. However, breakthrough cases of TT-CMV still occur. Despite improving the safety of blood components, testing remains a reactive approach to blood safety. In contrast, pathogen inactivation technologies offer a proactive approach with the potential to further improve blood safety. To reduce the risks associated with platelet transfusions, a photochemical treatment (PCT) process using a combination of the psoralen amotosalen HCl and long-wavelength UV light has been developed and introduced into clinical practice in Europe. PCT has been shown to result in greater than 5.9-log reductions in infectivity of human CMV in platelet concentrates and to prevent the transfusion transmission of murine CMV in a mouse transfusion model. Thus, PCT pathogen inactivation may play a role in further reducing the incidence of TT-CMV infection in patients who are at risk for serious CMV disease. Because PCT is a technology that targets nucleic acids, it also offers a proactive process for the inactivation of a broad range of viral, bacterial, and protozoan pathogens in addition to CMV. PMID- 16373188 TI - Clearance of prions during plasma protein manufacture. AB - Protein products isolated from human plasma are an important class of therapeutics that are used to treat patients afflicted with hereditary deficiencies, trauma, and severe infections. Because of the human origin of the starting material for the production of these biological products, there is a risk of transmitting infectious agents, including viruses and the infectious agents that cause transmissible spongiform encephalopathies (TSEs). The agent that is thought to cause TSEs is a disease-associated, misfolded form of the prion protein or PrP(Sc). Unlike viruses, there are no donor screening tests for TSEs available, and PrP(Sc) is resistant to traditional viral inactivation methods. Therefore, manufacturers of plasma products are faced with special challenges to ensure product safety with respect to TSEs. Fortunately, a growing body of evidence supports the capacity of manufacturing processes to remove infectious prions from the product stream during the purification of plasma products. This can be attributed in part to the unusual physicochemical nature of PrP(Sc), which is distinct from that of soluble therapeutic proteins. Although there is no reported TSE transmission through the use of plasma products to date, many unknowns remain to be addressed through long-term epidemiologic monitoring and further experimental studies. PMID- 16373189 TI - Proteomics: applications relevant to transfusion medicine. AB - With the completion of the human genome sequence, it is now possible to analyze the many individual components that comprise complex biologic systems. Despite this sequence data, understanding the biologic relationships of all proteins of a given cell or biologic sample (the proteome) is still an exceedingly difficult task. However, new technology developments mean that proteomics research can be used to investigate a variety of biologic systems. Already, these studies have given valuable insight for the development of improved diagnostic and therapeutic products. The present review aims to provide a basic understanding of proteomics research by discussing the methods used to study large numbers of proteins and by reviewing the application of proteomics methods to transfusion medicine. PMID- 16373190 TI - Electronic crossmatching. AB - The crossmatch (XM) is an important part of routine pretransfusion testing. It is used to detect ABO incompatibility and other clinically significant antibodies. The XM has been modified many times and, in recent years, has been abbreviated. The 2 common types of XM currently being done are the immediate spin XM for antibody-negative patients and the antiglobulin XM for the rest. The antiglobulin phase of the XM is generally considered optional, unless a clinically significant antibody is present. Originally, the XM was intended to be a final check for ABO compatibility and for the detection of unexpected antibodies. Electronic crossmatching was first introduced at the University of Michigan Medical Center in 1992. This paper summarizes the basic principles, current guidelines, regulatory requirements, and some recommendations that may prove useful for the implementation of electronic crossmatching. PMID- 16373191 TI - Kurt Stern (1909-2003). PMID- 16373192 TI - David B. Pall, PhD (1914-2004). AB - In this edition of the Pioneers and Pathfinders Series, the contributions of David B. Pall, PhD, to transfusion medicine are discussed. With the aid of Dr Pall's unpublished personal history and assistance from family members and the company he founded, we are able to provide perspective to several remarkable scientific advances. For those of us in transfusion medicine, the discovery of the world's first leukoreduction filter prevails as his most significant invention. However, to the rest of the world, Dr Pall pioneered filtration with applications in aerospace, microelectronics, general industry, and most recently, contamination control. The almost 60-year-old Pall Corporation continues to preserve his legacy. PMID- 16373193 TI - The road ahead with beta-blockers: expanding treatment options in cardiovascular disease. PMID- 16373194 TI - The role of beta-blockers as a cornerstone of cardiovascular therapy. AB - The beta-adrenergic blockade as a therapeutic approach first emerged in the 1950s. During the past five decades, the total number of indications that have been suggested, and the remarkable number approved by regulatory agencies, places beta-blockade far ahead of all competing treatments, not only in the cardiovascular area, but in all of therapeutics. Differentiation of beta adrenergic blocking agents has been made on the basis of beta1 selectivity, duration of action, intrinsic sympathomimetic activity, lipophilicity, and whether or not the beta-adrenergic blocking action is accompanied by an alpha adrenergic blocking action. With the development of nebivolol, a beta-adrenergic blocking agent that also activates nitric oxide synthase in blood vessels, comes a new therapeutic option. Endothelial dysfunction with loss of nitric oxide production is a common feature in many cardiovascular diseases. This fascinating class of drugs continues to provide us with new and important therapeutic opportunities. PMID- 16373195 TI - The role of the new beta-blockers in treating cardiovascular disease. AB - The beta-adrenergic receptor blockers play an important role in the management of cardiovascular disease, including hypertension and chronic heart failure. However, concerns regarding safety and tolerability with currently available agents can limit their use. The beta-blockers vary with regard to several pharmacologic properties, including beta1/beta2 selectivity, intrinsic sympathomimetic activity, and, with the newest beta-blockers, vasodilation. These pharmacologic differences may result in clinically important differences in tolerability and hemodynamic properties. Nebivolol is a novel beta-blocker with both a greater degree of selectivity for beta1-adrenergic receptors than other agents in this class and an ability to stimulate endothelial nitric oxide production, leading to vasodilation and other potential clinical effects. Published randomized, controlled, multicenter studies with nebivolol have shown that once-daily treatment significantly reduces systolic and diastolic blood pressure in patients with mild-to-moderate hypertension, compared with placebo, in a dose-dependent manner, and is well tolerated, with an adverse event profile similar to that of placebo. When compared with other beta-blockers as well as with other antihypertensive classes of agents in head-to-head trials, nebivolol demonstrated similar antihypertensive efficacy and a lower incidence of adverse events. Nebivolol has also been shown to significantly reduce morbidity and mortality in a large population of elderly patients with chronic heart failure, independent of left ventricular ejection fraction. Nebivolol is currently available in Europe for the management of hypertension and is expected to be available soon in the United States. PMID- 16373196 TI - Exploring vascular benefits of endothelium-derived nitric oxide. AB - Although the regulation of arterial blood flow has been a subject of intensive medical research, the precise circulatory mechanisms involved are still not fully understood. It has been increasingly recognized that the endothelium plays a vital role in regulating vascular tone, structure, and function. A seminal discovery was made with the identification of endothelium-derived relaxing factor, a key mediator of vasodilation, which was later identified as nitric oxide (NO). Nitric oxide is synthesized from the amino acid L-arginine in the endothelium. Decreased bioavailability of NO is associated with arterial stiffness, hypertension, atherosclerosis, and cardiovascular disease (CVD). Nebivolol is a novel beta-blocker that is highly selective for beta1-adrenergic receptors. Nebivolol also causes vasodilation through a mechanism involving endothelium-derived NO. In clinical studies in hypertensive subjects, nebivolol significantly improves vasodilator responses to endothelium-dependent agonists such as acetylcholine. In addition, nebivolol significantly reduces pulse wave velocity (PWV), a measure of arterial stiffness, whereas the beta-blocker atenolol has no effect on PWV. Because endothelial dysfunction and arterial stiffness play an integral part in the early atherosclerotic process and are associated with poor outcomes and increased mortality, independent of blood pressure, the ability of nebivolol to enhance release of endothelium-derived NO may have significant clinical implications for the use of this agent in the treatment of hypertension and CVD. PMID- 16373197 TI - Towards the world-wide ban of indoor cigarette smoking in public places. AB - In 1984, Dr. C. Everett Koop, then Surgeon General of the US, presented an important speech on the hazards of smoking. In his speech, he stated "The ultimate goal should be a smoke-free society by the year 2000." In addition, the World Health Organization (WHO) has initiated a process to ban smoking globally; on 21 May 2003, at the 56th World Health Assembly, WHO's 192 Member States unanimously adopted the world's first public health treaty, the WHO Framework Convention on Tobacco Control. Although good progress has been made, reaching the ultimate goal is far from certainty. Therefore, it is time to re-visit this crucial public health activity and re-energize the effort to reach this goal. Since numerous reports have been written on the ban of smoking based on benefits to the smokers, the emphasis of our report is on benefits to non-smokers from their exposure to environmental tobacco smoke (ETS). We provide a concise review of the impact of ETS on health and economy. In addition, we examined the different interest groups on supporting and opposing the ban, the role of the government, private citizens and medical professionals on this activity, and certain constraints on implementing the global ban. We also provide some recommendations on how to promote the ban globally. Since cigarette smoking is an unnecessary habit that has devastating consequences around the world, banning of cigarette smoking should be a global mission. A global ban on indoor smoking in public places is an important first step in an international effort to prevent morbidity and mortality caused by tobacco smoking and ETS. PMID- 16373198 TI - Cytogenetic effect of technical glyphosate on cultivated bovine peripheral lymphocytes. AB - A technical herbicide containing isopropyl amine salt of glyphosate was tested for induction of chromosome aberrations (CA) and sister chromatid exchanges (SCE) in cultured bovine peripheral lymphocytes. Cultures were exposed to a glyphosate formulation at concentrations ranging from 28 to 1120 micromol/l without and with metabolic activation. No clastogenic effect of the herbicide was found. Its genotoxic effect was confirmed in the SCE assay after 24 h of incubation. A statistically significant elevation in SCE induction was observed in each of the donors after application of the product at doses ranging from 56 to 1120 micromol/l. The highest concentrations (560 and 1120 micromol/l) also caused reduction of mitotic and proliferation indices. In the 2 h-assay with metabolic activation a statistically significant frequency of SCE was observed only in cultures treated with the agent at a concentration of 140 micromol/l. PMID- 16373199 TI - Polymorphism in DNA repair genes and oral squamous cell carcinoma in Thailand. AB - DNA repair capacity is essential in maintaining cellular functions and homeostasis. However, the repair capacity can be altered based on DNA sequence variations in DNA repair genes and thus may cause cancer susceptibility. We investigated associations between polymorphisms in DNA repair genes and oral squamous cell carcinoma (OSCC) in a Thai population. Nine known single nucleotide polymorphisms (SNPs) in five common DNA repair genes were investigated: XRCC1 (Arg194Trp and Arg399Gln); XRCC3 (Thr241Met); XPC (PAT and Lys939Gln); XPD (exon 6, and Lys751Gln); and MGMT (Trp65Cys and Leu84Phe). We studied 106 cases and 164 healthy controls that were frequency-matched by age (+/-5 years), gender, and cigarette smoking and alcohol drinking habits. The genotype assays were performed using the polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) method. The R version 2.0.1 statistical software was applied for statistical analysis of association. Based on multivariate analyses, we found that the variant genotypes with XRCC3 241Met exhibited a >3-fold elevated risk (OR = 3.3, 95% CI = 1.31-8.36, p = 0.01) for OSCC. There was a marginally significant risk observed in variants with XRCC1 194Trp (OR = 1.81, 95% CI = 0.91 3.63, p = 0.09) and XPD exon 6 (OR = 1.71, 95% CI = 0.93-3.16, p = 0.09). Combination of the variant genotypes of these three susceptibility genes was associated with a highly significant risk for OSCC (OR = 9.43, 95% CI = 1.98 44.9, p < 0.01). From further multivariate analyses, the variants with XRCC1 194Trp and possibly XRCC3 241Met interacted with tobacco and alcohol to further increase the risk (OR = 3.37 95% CI = 1.41-8.02, p < 0.01; OR = 2.92, 95% CI = 0.94-9.04, p = 0.06). On the other hand, increased risk was detected in non-betel chewers (OR = 2.88, 95% CI = 1.31-6.31, p < 0.01; OR = 2.61, 95% CI = 0.97-7.11, p = 0.06) who carry the two variant genotypes, respectively. Males with the variants XRCC1 194Trp or XRCC3 241Met had a higher risk of developing OSCC than males with the corresponding wild-type genotypes (OR = 2.72, 95% CI = 1.34-5.52, p < 0.01; OR = 2.95, 95% CI = 1.12-7.75, p < 0.05). Such association was not detected in females. Interestingly, the risk increased in female carriers of XPD exon 6 (OR = 3.93, 95% CI = 1.14-13.6, p < 0.05). We could not demonstrate a significant interaction of these SNPs with age in this study. Our data indicate that the variant genotypes with XRCC3 241Met and possibly XRCC1 194Trp and XPD exon 6 contribute to OSCC development in a Thai population. In addition, these SNPs influence the repair of DNA damage that is caused by environmental risk factors for oral cancer. PMID- 16373200 TI - Environmental exposure to residential pesticides, chemicals, dusts, fumes, and metals, and risk of childhood germ cell tumors. AB - We examined relationships between exposure to residential pesticides, chemicals, dusts, fumes, and metals, and childhood germ cell tumors (GCTs) in the largest case-control study to date on the topic. We recruited 272 children under 15 years old who had GCT diagnosed between January 1, 1993 and December 31, 2001. Controls were selected by random-digit dialing and were frequency matched to cases by sex, age, and geographic area. Telephone interviews and self-administered questionnaires of parents were used to collect exposure information. We used unconditional logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI). Maternal and paternal exposure from 6 months before pregnancy to during breastfeeding and children's postnatal exposure to residential pesticides were generally unrelated to risk of childhood GCT. Elevated OR were observed for mothers' exposure to hair dyes 1 month before pregnancy for boys (OR = 1.7, 95% CI = 1.0-2.8) and during breastfeeding for boys and girls combined, and for girls (OR = 1.5, 95% CI = 1.0-2.2 and OR = 1.7, 95% CI = 1.1-2.6, respectively). Among boys, OR for paternal exposure to insecticides more often than four times or exposure to indoor insecticides more often than three times were 0.5 (95% CI = 0.2-0.9, trend test p = 0.05) and 0.3 (95% CI = 0.2-0.8, trend test p = 0.02) during the index pregnancy. Overall this study produced no strong evidence linking parental and child residential exposure to pesticides, certain chemicals, dusts, fumes, and metals to increased risk of childhood GCT. Statistically significant associations need to be confirmed in future studies. PMID- 16373201 TI - Aircraft disinsection: exposure assessment and evaluation of a new pre embarkation method. AB - A new "pre-embarkation" method for aircraft disinsection was investigated using two different 2% d-phenothrin containing aerosols. Five experiments in aircrafts of the type Airbus 310 (4x) and Boeing 747-400 (1x) were performed. In the absence of passengers and crew the d-phenothrin aerosol was sprayed under the seat rows and in a second step at the height of approximately 1.60 m by moving from one end of the cabin to the other. Concentration levels of d-phenothrin were determined at different time periods after application of the aerosol spray. In a B 747-400 with the air conditioning system operating the concentrations ranged between 853 and 1753 microg/m3 during and till 5 min after the beginning of spraying at different locations in the cabin. Within 5-20min after the end of the spraying concentrations of 36-205 microg/m3 and 20-40 min thereafter only ca. 1 microg d-phenothrin/m3 were detectable (average values in relation to each period of measurement). On cabin interior surfaces the median values for mainly horizontal areas ranged from 100 to 1160 ng d-phenothrin/cm2. d-Phenothrin concentrations in the air were sufficient to kill flying insects like house flies and mosquitoes within 20 min. Horizontal surfaces were 100% effective against insects up to 24 h after spraying. Doses inhaled by sprayers determined by personal measurements were calculated to be 30-235 microg d-phenothrin per 100 g spray applied (30% in the respirable fraction for Arrow Aircraft Disinsectant; 10% for Aircraft Disinsectant Denka). If passengers will board, e.g., 20 min after the end of the disinsection operation, inhalation exposure is estimated to be practically negligible. Also possible dermal exposure from residues in seats and headrests is very low for passengers during the flight. Therefore any health effects for passengers and crew members are very unlikely. PMID- 16373202 TI - Wilms' tumor and exposures to residential and occupational hazardous chemicals. AB - This case-control study examines the association between residential and occupational exposures to hazardous chemicals and the risk of Wilms' tumor. The study included 303 cases recruited from six state cancer registries, who were diagnosed between January 1, 1992 and December 31, 1995. A total of 575 controls selected through random digit dialing were frequency matched to the cases. A standard questionnaire was administered to participants during a telephone interview. Parental residential addresses and locations of US Environmental Protection Agency National Priority List (NPL) sites were geocoded and analyzed, along with occupational exposure information. There were no cases of Wilms' tumor found in individuals living within one-half mile distance of a hazardous waste site. However, elevated odds ratios were found for using hairdressing chemicals, motor oil, paint, paint stripper, and pesticides during the pregnancy term and during the 2-year period prior to birth. The findings do not support the hypothesis that Wilms' tumor is associated with residing near an NPL site. PMID- 16373203 TI - Health complaints and annoyances after moving into a new office building: a multidisciplinary approach including analysis of questionnaires, air and house dust samples. AB - After moving into a new office building, employees complained about eye irritations, sore throats and unspecific symptoms. They were concerned about visible dust as a potential cause of adverse health effects. An external working group was appointed to investigate indoor air pollution and health complaints and to suggest measures to improve the situation. Air samples and floor dust samples for analysis of organic compounds were collected in three offices. Bimonthly during 8 months, measurement campaigns were conducted to assess the trend of air pollutants. A questionnaire was administered concerning environmental conditions at the work place and complaints before and after moving into the new office building. Overall the concentrations of volatile organic compounds (VOC) and formaldehyde were fairly low. However, initially high concentrations (4300-7800 mg/kg) of tris-(2-butoxyethyl)-phosphate (TBEP) and diethylhexylphthalate (DEHP: 980-3000 mg/kg) were found in dust samples. The coating of the rubber floor was identified as the source of TBEP, while no single predominant source of DEHP was found. Results of the questionnaire demonstrated an increased irritation of the mucous membranes and a reduction of well-being after the employees had moved into the new building. Perception of low relative humidity and high temperature as well as unpleasant odors were associated with respiratory complaints. After removal of the coating of the rubber floor throughout the whole building, a reduction up to 90% of TBEP in the dust samples was found. In spite of several attempts, no such marked reduction was achieved with the concentration of phthalates. Although there was no significant association between visible dust exposure and increase of complaints after moving in, room climate conditions that could increase the deposition of dust in the airways were associated with the complaints. Hence it cannot be ruled out that fine dust containing TBEP together with unfavorable indoor factors were responsible for the development of the complaints. PMID- 16373204 TI - Body weight and water ingestion estimates for women in two communities in the Philippines: the importance of collecting site-specific data. AB - In this paper, we report data on exposure factors relevant to assessing health risks of contaminant exposures to women of childbearing age (age 15-49) in two communities in the Philippines. We collected exposure factor data through an interview survey of 182 women conducted between January and May 2002 and we present distributions of self-reported body weight and water ingestion rates. A simple comparison of our results to those from large studies of US women suggests that these small subpopulations may both weigh less and consume more water than might be expected based on the extensive national US data, and it suggests that exposure analysts focused on a particular geographic area should consider the value of obtaining site-specific data to characterize exposure and risk. The lack of a comparable large study of water ingestion rates and body weights of Filipino women makes it difficult to determine whether the communities studied are typical of the population of the Philippines or represent unique subgroups. We suggest that Philippine health authorities consider the possibility of including questions about water ingestion rates in future national health or nutrition surveys. PMID- 16373205 TI - Detection of genetically modified DNA sequences in milk from the Italian market. AB - The possible transfer and accumulation of novel DNA and/or proteins in food for human consumption derived from animals receiving genetically modified (GM) feed is at present the object of scientific dispute. A number of studies failed to identify GM DNA in milk, meat, or eggs derived from livestock receiving GM feed ingredients. The present study was performed in order to: (i) develop a valid protocol by PCR and multicomponent analysis for the detection of specific DNA sequences in milk, focused on GM maize and GM soybean; (ii) assess the stability of transgenic DNA after pasteurization treatment and (iii) determine the presence of GM DNA sequences in milk samples collected from the Italian market. Results from the screening of 60 samples of 12 different milk brands demonstrated the presence of GM maize sequences in 15 (25%) and of GM soybean sequences in 7 samples (11.7%). Our screening methodology shows a very high sensitivity and the use of an automatic identification of the amplified products increases its specificity and reliability. Moreover, we demonstrated that the pasteurization process is not able to degrade the DNA sequences in spiked milk samples. The detection of GM DNA in milk can be interpreted as an indicator of fecal or airborne contamination, respectively, with feed DNA or feed particles, although an alternative source of contamination, possibly recognizable in the natural environment can be suggested. Further studies, performed on a larger number of milk samples, are needed to understand the likely source of contamination of milk collected from the Italian market. PMID- 16373206 TI - Influence of growth media on the sensitivity of Staphylococcus aureus and Pseudomonas aeruginosa to cationic biocides. AB - In this study, the influence of culturing Staphylococcus aureus and Pseudomonas aeruginosa under different growth conditions on their inactivation by the cationic active compounds benzalkonium chloride, chlorhexidine digluconate and octenidine dihydrochloride was investigated. Cells were grown in non-agitated tryptone soya broth as well as on tryptone soya agar according to national and international standards for evaluating chemical disinfectants. In quantitative suspension tests, cells of both test organisms grown on agar were significantly more sensitive to all three biocides than cells grown in broth. The differences in antimicrobial activity were greater in the case of S. aureus than in the case of P. aeruginosa. With S. aureus cultures, differences in the reduction factor of up to 5 log steps were found, with P. aeruginosa up to 2.5 log steps. The results of our uptake tests performed with S. aureus and octenidine dihydrochloride indicated that the growth conditions and the associated different stress factors either had an influence on the composition of the cell surface of this test organism or induced the formation of an efflux system. Cells of S. aureus cultured in broth took up only one-fifth of the amount of biocide molecules compared to cells from agar cultures. These data correlated with the results of the suspension tests. A low uptake of biocides apparently led to a reduced killing rate. In contrast to S. aureus, no significant differences in the uptake of octenidine dihydrochloride by cells of P. aeruginosa could be observed. These cells took up the same amount of the antimicrobial substance, whether on agar or in broth. In view of these results, possible consequences should be considered prior to changing test regulations. PMID- 16373207 TI - Rotaviral RNA found in wastewaters from hospital laundry. AB - The aim of this prospective study was to determine the presence of rotaviral RNA in water from a hospital laundry. Since rotaviruses are known as major causal agents of diarrhoea in humans, it is necessary that laundering hospital textiles results in efficient chemo-thermal disinfection, thus minimizing the possibility of transmission of rotaviruses to immune-compromised patients in hospitals. RT PCR and second round PCR for gene amplification using specific primers, succeeding ultra-filtration and RNA isolation, was used to determine the presence of rotaviral RNA in water samples. The results show that rotaviral RNA was found in wastewater after the washing process, thus confirming an inadequate disinfecting effect of the examined laundering procedures. PMID- 16373208 TI - Isolation of Salmonella serotypes in wastewater and effluent: Effect of treatment and potential risk. AB - The characteristics of strains of Salmonella isolated in wastewater were compared before and after treatment with activated sludge (conventional sewage treatment). Of the 21 strains studied, 11 came from raw wastewater and 10 from treated water. Our objective was to determine if the treatment affected all serotypes to the same extent, and whether it had an effect on the plasmidic composition and selection of resistances to antibiotics and disinfectants. The serotypes of all strains were identified, and we also determined the sensitivity to antibiotics, the minimal bactericidal concentration of three disinfectants (peracetic acid, glutaraldehyde, and sodium hypochlorite), and plasmidic content. Results showed no significant differences in the isolated strains before versus after treatment, thus indicating that the conventional treatment of sewage does not constitute a risk factor in the selection of Salmonella strains with a greater pathogenic potential. PMID- 16373209 TI - Evaluation, evidence and effectiveness. PMID- 16373210 TI - Information sharing and children's services reform in England: can legislation change practice? AB - The Laming Report into the death of Victoria Climbie reiterated the long-standing critique of inadequate communication and coordination amongst the key professions and agencies. It led directly to the Green Paper, Every Child Matters in 2003 and the subsequent Children Act 2004. Amongst other things the Act proposes the establishment of a database on every child, which would be accessible to a range of practitioners - a measure that has been hotly contested. This article examines the reasons for this contestation and explores the extent to which a compromise might be judged to have dislodged a key policy objective. PMID- 16373211 TI - Health professionals in multi-disciplinary and multi-agency teams: changing professional practice. AB - The article draws on an Economic and Social Research Council (ESRC)-funded research project that aimed to investigate the reality behind the rhetoric of "joined up thinking". The research project was a qualitative, multi-method study involving three phases, including observation and documentary analysis; interviews; and focus groups around decision making and knowledge sharing. The article reflects on the perspectives and experiences of health professionals and their colleagues in multi-agency teams about the impact of multi-agency teamwork on their professional knowledge and learning, and on their ways of working. Actual and potential conflicts between professionals are explored about models of understanding, about roles, identities, status and power, about information sharing, and around links with other agencies. Dilemmas of team building and of conflicting values and knowledge are exemplified from health professionals' accounts, using theoretical models of "communities of practice" and "activity theory". The article presents groups of strategies that health professionals and their colleagues in multi-agency, multi-professional teams use to overcome barriers and to strengthen team cohesion. The conclusion reflects on some implications of our findings in theory and practice for professionalism within integrated, multi-professional teams that are building new ways of working. PMID- 16373212 TI - Palliative care teams: effective through moral reflection. AB - Working as a multidisciplinary or interdisciplinary team is an essential condition to provide good palliative care. This widespread assumption is based on the idea that teamwork makes it possible to address the various needs of the patient and family more effectively. This article is about teamwork and about the effectiveness of teams working in palliative care. First, the nature of teamwork will be highlighted. Second, attention will be paid to team effectiveness; what exactly is team effectiveness and with what parameters can it be measured? Third, the nature of moral reflection and moral deliberation in palliative care will be highlighted. A concrete process of moral deliberation will be described. In conclusion, we shall argue that the capacity for moral reflection is a feature of a team working effectively. PMID- 16373213 TI - Interprofessional postgraduate education in primary health care: is it making a difference? AB - This paper explores attitudes to, and perceptions of, the impact of interprofessional postgraduate education for primary health care professionals, based on a postal survey of 153 primary health care professionals undertaking postgraduate qualifications in New Zealand. The response rate was 75% (114/153 responses); comprising 79 doctors, 28 nurses, 7 other health professionals. As a result of their postgraduate education, 92% (104/113) reported improvement in their own practice; 68% (72/106) reported a positive influence on their workplace practice. Forty-eight percent (53/111) increased their understanding of their own professional role; 79% (77/98) increased their understanding of another professional groups' skills and competencies. Twenty-two percent (25/114) perceived increased career opportunities within a year; 56% (64/114) in the longer term. Only 12% (14/114) perceived future increases in income as a result of their study. Interprofessional postgraduate qualification study for primary health care professionals in New Zealand resulted in personal and professional benefit for individuals and their clinical practice, and increased understanding about their own and other health professionals' roles. The interprofessional nature of the education was seen as positive, contributing to a modest increase in collaboration between health professional groups. Barriers to furthering participation in interprofessional learning and increasing intersectorial collaboration in the workplace are identified and discussed. PMID- 16373214 TI - Australian evidence for interprofessional education contributing to effective teamwork preparation and interest in rural practice. AB - A pilot interprofessional education (IPE) placement for undergraduate health care professional students was undertaken in rural Victoria, Australia from 2001 to 2003. Medical, nursing, physiotherapy and pharmacy students were involved, and the project is ongoing. This paper briefly outlines the educational model, then focuses on the evaluation methods and results obtained from student evaluations. The placement experience improved self-reported teamwork skills and knowledge, and supported participating students' belief in the value of interprofessional practice. Placements strengthened nursing and allied health students' intention to work in rural health settings after graduation. The rural interprofessional educational experience improved interprofessional abilities in a group of students who have the potential to influence change towards collaborative practices in their future workplaces. The results obtained provide sufficiently strong evidence to justify the continuation and expansion of this educational model in the Australian setting. Pedagogical and evaluation modifications are discussed that may benefit future IPE programs. PMID- 16373215 TI - The Readiness for Interprofessional Learning Scale: a possible more stable sub scale model for the original version of RIPLS. AB - The original version of the Readiness for Interprofessional Learning Scale (RIPLS) was published by Parsell and Bligh (1999). Three sub-scales with acceptable or high internal consistencies were suggested, however two publications suggested different sub-scales. An investigation into how to improve the reliability for use of the RIPLS instrument with undergraduate health-care students commenced. Content analysis on the original 19 items involving experienced health-care staff resulted in four sub-scales. These sub-scales were then used to formulate a possible model within a structural equation model. The goodness of fit was assessed using a sample (n = 308) of new first year undergraduate students from 8 different health and social care programmes. The same data was fitted to each of the two original sub-scale models suggested by Parsell and Bligh (1999) and the results compared. The fit of the new four sub scale model appears superior to either of the original models. The new four factor model was then tested on subsequent data (n = 247) obtained from the same students at the end of their first year. The fit was seen to be even better at the end of the academic year. PMID- 16373216 TI - An e-resource to coordinate research activity with the Readiness for Interprofessional Learning Scale (RIPLS). AB - Research into interprofessional education is often performed by professionals in practice alongside their other "core" duties, thus additional help and support to achieve quality research outcomes would be beneficial. This study aimed to create a virtual community of researchers with shared interest in quantitative studies of interprofessional learning using the Readiness for Interprofessional Learning Scale. An e-resource was built which provided access to relevant resources, helped to coordinate research effort, and promoted communication. Use of the resource was monitored over a 14-month period and evaluated at the end of this time. Forty-eight researchers across six different countries were assigned usernames and passwords. According to a self-report measure, 23/27 (85%) respondents to the evaluation questionnaire had accessed the resource and 95% planned to use it in the coming year. This paper shares our experiences and observations from the study. PMID- 16373217 TI - Methodological understandings and misunderstandings in interprofessional research: experiences of researching transitional rehabilitation for older people. AB - Drawing on our shared experience of a mixed-methods evaluation of a rehabilitation scheme for older people in six residential care homes in Nottingham, this paper examines the challenges and rewards of interprofessional research. A social anthropologist, social work academic and medical academic respectively, we explore understandings and misunderstandings that arose during the two-year research process as we sought to integrate quantitative and qualitative findings from a randomized controlled trial (RCT) and an ethnographic study conducted in parallel. We identify ways in which it was possible to overcome the danger of fragmentation, which we argue may be inherent in interprofessional research. This includes a recognition that researchers from different disciplinary backgrounds may use different "key symbols" to view the world but that professional knowledge and expertise may enable researchers to transcend such disciplinary boundaries and to hear what each is saying to the other for the benefit of improved collaboration. PMID- 16373223 TI - Lymphoproliferative disorders. PMID- 16373224 TI - The epidemiology of non-Hodgkin lymphoma. AB - Non-Hodgkin lymphoma (NHL) includes a group of more than 20 different malignant lymphoproliferative diseases that originate from lymphocytes. Rates of NHL have increased dramatically over the past few decades, although the rate of increase has recently slowed. It is now the sixth most common cancer in Australia. Globally, it is somewhat more common in men than in women, and rates are highest in North America and Australia. The causes of the increase in NHL rates are largely unknown. The best described risk factor for NHL is immune deficiency; rates of NHL are greatly increased, with relative risks of 10-100 or more, in people with immune deficiency associated with immune suppressive therapy after transplantation, HIV/AIDS, and congenital conditions. In addition, some NHL subtypes are associated with specific infections. These include immune-deficiency associated central nervous system NHL (Epstein-Barr virus); gastric mucosa associated lymphoid tissue NHL (Helicobacter pylori); adult T-cell leukemia/lymphoma (human T-lymphotrophic virus type 1) and body cavity-based lymphoma (human herpesvirus 8). However, these specific infections account for a very small proportion of total NHL incidence. In addition to immune deficiency and infection, other immune-related conditions are increasingly being recognised as related to NHL risk. Specific autoimmune conditions, including rheumatoid arthritis, systemic lupus erythema, Sjogren's syndrome, psoriasis and coeliac disease are associated with moderately increased risk of NHL. On the other hand, allergic and atopic conditions and their correlates such as early birth order, appear to be associated with a decreased risk of NHL.A variety of other exposures are less strongly related to NHL risk. These include occupational exposures, including some pesticides, herbicides, and solvents. Recently, two studies have reported that sun exposure is associated with a decreased risk of NHL. Smoking appears to be weakly positively associated with risk of follicular NHL, and alcohol intake is associated with a decreased risk of NHL. The pooled analysis of several case-control studies of NHL risk that are currently in the field promises to help clarify which of these risk factors are real, and will contribute to the elucidation of the mechanisms of how disorders of the immune system, and other factors, are related to NHL risk. PMID- 16373225 TI - Viruses and lymphoma. AB - The aetiology of lymphomas is poorly understood and the striking increase in its incidence rate in developed societies remains unexplained. The concept of lymphoma as a virally-induced malignancy is not surprising since viruses are implicated in approximately 15% of all cancers. However, lymphoma represents a complex multistep process and, although viral associations have been identified, integration of the available epidemiological and scientific data poses substantial questions. The study of oncogenic viruses has and will continue to yield major insights into the pathogenesis of lymphoma. Further research is likely to uncover new lymphoma associations between both known and as yet unidentified viruses, may provide cellular and pharmacological targeted antiviral therapy strategies for the treatment of malignant lymphoma, and ultimately may generate the most promising avenue for lymphoma prevention. PMID- 16373226 TI - Diagnostic pathology of lymphoproliferative disorders. AB - The last 20 years have seen a dramatic change in the way we classify, and therefore diagnose, lymphoma. Two decades ago, the International Working Formulation enabled diagnosis and management on the basis of H&E sections alone, with no mandatory requirement for immunophenotyping, molecular studies or any other ancillary investigations. The concept of categorisation by 'clinicopathological entities' defined by clinical features, morphology, immunophenotype and more recently, genotype, began with the Kiel, and Lukes and Collins classifications in the late 1970s, becoming fully expressed in the REAL and subsequently WHO classifications. The current, multidisciplinary approach to categorisation adds significantly to the task facing the anatomical pathologist, since it requires distribution of biopsy material to all the appropriate specialised laboratories, the gathering of a range of cross-disciplinary information, the correlation of all diagnostic findings, deduction of a definitive diagnosis and, finally, integration of all the above into a single multiparameter report. In this review, we summarise the contemporary approach to the biopsy, diagnosis and reporting of lymphoproliferative disorders. PMID- 16373227 TI - Immunophenotyping of lymphoproliferative disorders: state of the art. AB - Immunophenotyping was introduced into diagnostic pathology over 30 years ago to assist in the diagnosis and classification of lymphoproliferative disorders. Today the role of immunophenotyping has been expanded beyond this to include the detection of markers of prognosis, determination of disease phenotypes associated with specific chromosomal abnormalities, detection of targets for immunotherapy and to monitor residual disease. Immunoperoxidase detection methods remain the most popular in histopathology, whilst flow cytometry is most commonly applied for haematological samples. The range of monoclonal antibodies available, including those which work in routinely performed tissue specimens, continues to increase. This is in part a result of gene expression studies identifying precise genetic signatures for certain lymphoproliferative disorders and the generation of new protein markers to gene products of upregulated genes. This review summarises the current status and applications of immunophenotyping in the assessment of many of the lymphoid malignancies. PMID- 16373228 TI - Organisation of neoplastic haematopathology services: a UK perspective. AB - The diagnosis of haematological malignancies has begun to emerge as a distinct pathological discipline in the United Kingdom. This has been driven by the recommendation of the National Institute for Clinical Excellence that diagnosis of leukaemia and lymphoma should take place in a specialist laboratory and in most cases this should be organised on a regional basis. The reason for this guidance was the perception that there was a considerable level of diagnostic inaccuracy and that this could be improved by better integration of the currently available technologies. This is one of a number of major changes in the way that services to patients are being delivered, all of which are centred on the development of multidisciplinary teams responsible for the provision of local services. The introduction of the WHO classification of haematological malignancy provides a structure for the development of integrated haemtopathology laboratories, with its emphasis on definition of disease entities based on clinical, morphological, phenotypical and molecular features. This means that these diagnostic modalities can be used systematically and in parallel to provide effective cross validation of a diagnosis. One of the challenges raised by this approach is the selection of the most informative panels of investigations both at presentation and subsequent follow up from the wide range of options that are now available. The introduction of specialist haematopathology services in the United Kingdom has highlighted a number of scientific and organisational issues that in time may have a wider impact on diagnostic laboratories in general. These include the relationship between size and cost effectiveness and the future role of clinical scientists and medically trained pathologists. Integrated laboratories of the type being developed challenge the prevailing model for delivery of pathological services in the United Kingdom, which is based around the traditional pathology disciplines. These speciality boundaries will become less relevant as long established diagnostic techniques are replaced by the new generation of diagnostic technologies and it is important to establish frameworks of service delivery that can deploy these developments for the benefit of patients. PMID- 16373229 TI - Cytogenetics of lymphomas. AB - Cytogenetic analysis is now a routine part of the diagnosis and management of a significant number of lymphoid malignancies. Whilst conventional cytogenetics remains the most comprehensive method for assessing chromosome abnormalities, the technical difficulties associated with conventional cytogenetics in most lymphomas has resulted in increased use of fluorescence in situ hybridisation (FISH) to identify specific abnormalities that are useful in either the diagnosis or management of these disorders. The finding of one of the Burkitt's translocations is of major importance in the diagnosis of Burkitt's and Burkitt's like lymphomas, whereas the t(14;18), although seen in most follicular lymphomas (FL), is not usually required to make a diagnosis. Thus, whilst cytogenetics may be of interest in FL, it is not an essential part of the diagnostic work-up. Conventional cytogenetics may be useful for identifying markers of resistance to Helicobacter pylori therapy in MALT lymphomas. In disorders such as Hodgkin lymphoma, hairy cell leukaemia and lymphoplasmacytoid lymphoma, although many cytogenetic abnormalities have been observed, no consistent or specific abnormalities have been identified and so, at this point in our knowledge of the genetics of these disorders, cytogenetics cannot be considered a useful test for either diagnosis or prognosis. In contrast, the diagnosis of mantle cell lymphoma is now dependent upon the identification of the 11;14 translocation that results in cyclin D1 up-regulation. It is widely acknowledged that FISH is the most consistently useful test to identify the juxtaposition of the CCND1 and IGH genes in mantle cell lymphoma and is regarded as the 'gold standard'. FISH also has a role in identifying genetic abnormalities of prognostic significance in chronic lymphocytic leukaemia. Given the wealth of genetic and cytogenetic abnormalities that are continuing to be found in chronic lymphoid malignancies, it will be some time before the optimal use of both conventional cytogenetics and FISH is established in the diagnosis and management of lymphomas. PMID- 16373230 TI - Molecular biology of lymphoma in the microarray era. AB - This review will focus on the molecular biology of lymphoproliferative disorders with emphasis on lymphomas. The spectrum of known recurrent gene rearrangements found in lymphomas will be outlined and their relevance to diagnosis and subclassification of disease will be discussed. Finally, a survey of the current trends in gene expression profiling of lymphomas by microarray technology will be presented with reference to implications for diagnosis, classification, prognosis and treatment. PMID- 16373231 TI - Lymphoproliferative disorders: prospects for gene therapy. AB - The lymphoproliferative disorders represent a large group of diseases with a significant variation in presentation and clinical course. There has been a trend of increasing incidence for some of these disorders, and despite advances in therapies, a significant number of patients either respond poorly or have early relapses. For this reason there is a need to investigate novel therapies to be used either alone or as adjunct treatment in combination with conventional therapies. Gene therapy is a relatively new field that takes advantage of our increased understanding of molecular biology with the aim of treating a variety of diseases including cancer. It is defined as the introduction of genetic material into cells for therapeutic intent. Methods to improve gene delivery efficiency have been the focus of a large amount of research and to date the optimal procedure uses viruses such as oncoretroviruses, lentiviruses, adenoviruses, adeno-associated viruses and herpes simplex viruses. There are four main gene therapy strategies that might be used for the treatment of lymphoproliferative disorders. First, immunotherapy using tumour vaccines or techniques to enhance the function of immune effector cells has been investigated with some success in patients with B-cell malignancies. Second, the introduction of prodrug-activated 'suicide' genes into cells has been explored, in particular in patients with post-transplantation lymphoproliferative disease. Third, direct lysis of tumour cells using viruses shows some early promise, especially in the treatment of B-cell disorders by manipulating the measles virus to target the CD20 antigen. Finally, anti-gene strategies such as anti-sense therapy, ribozymes, and most recently RNA interference, could be used to suppress expression of specific target genes. RNA interference in particular has tremendous potential and has been studied in the context of anaplastic large cell lymphoma as well as Epstein-Barr virus-associated malignancies. Whilst we are still in the early days of this field and to date results have been modest, there is still a significant potential for gene therapy to play a role in the future treatment of these disorders. PMID- 16373232 TI - Vaccine strategies to treat lymphoproliferative disorders. AB - Lymphoproliferative disorders, including follicular lymphoma (FL), multiple myeloma (MM) and chronic lymphatic leukaemia (CLL), are slowly progressive malignancies which remain incurable despite advances in therapy. Harnessing the immune system to recognise and destroy tumours is a promising new approach to treating these diseases. Dendritic cells (DC) are unique antigen-presenting cells that play a central role in the initiation and direction of immune responses. DC loaded ex vivo with tumour-associated antigens and administered as a vaccine have already shown promise in early clinical trials for a number of lymphoproliferative disorders, but the need for improvement is widely agreed. Recent advances in the understanding of basic DC biology and lessons from early clinical trials have provided exciting new insights into the generation of anti tumour immune responses and the design of vaccine strategies. In this review we provide an overview of our current understanding of DC biology and their function in patients with lymphoproliferative disorders. We discuss the current status of clinical trials and new approaches to exploit the antigen presenting capacity of DC to design vaccines of the future. PMID- 16373233 TI - Is haemolytic staining of the aortic root a sign of fresh water drowning? PMID- 16373234 TI - Intraosseous embryonal rhabdomyosarcoma as a second neoplasm following retinoblastoma. PMID- 16373235 TI - Primary transitional cell carcinoma of the endometrium. PMID- 16373236 TI - Adenomyoepithelioma of the breast with squamous and sebaceous metaplasia. PMID- 16373237 TI - Juvenile temporal arteritis: a case report. PMID- 16373238 TI - Basaloid squamous cell carcinoma versus basal cell adenocarcinoma of the oral cavity. PMID- 16373239 TI - Autosomal recessive adult-onset hypophosphatasia. PMID- 16373240 TI - Screening for colorectal cancer using faecal blood testing: varying the positive cut-off value. PMID- 16373241 TI - Neuroendocrine carcinoma of the prostate gland: authors' reply. PMID- 16373242 TI - Drospirenone in the treatment of severe premenstrual cerebral edema in a woman with antiphospholipid syndrome, lateral sinus thrombosis, situs inversus and epileptic seizures. AB - We report herein the case of 32-year-old woman with situs inversus, thrombophilia, antiphospholipid syndrome and severe premenstrual syndrome (PMS) with cerebral edema and epileptic seizures prior to menstruation. Seven days prior to regular menstruation she developed severe PMS, including headache, blurred vision, epileptic seizures, urinary incontinence, craving for food, depression and irritability. Papilledema was detected. Daily hormone analyses prior to and during menstruation confirmed an ovulatory cycle with extremely high progesterone, prolactin and insulin levels in the late luteal phase. From day 29 to day 31, progesterone and insulin decreased sharply and the estradiol/progesterone ratio changed, leading to epileptic seizures and the peak of her symptoms. Diuretic treatment was administered. All symptoms disappeared during the first few days of menstruation. A novel oral contraceptive, containing ethinyl estradiol and drospirenone, an antimineralocorticoid progestogen, was given during the next cycle and hormone analyses were repeated. All symptoms were reduced significantly and no cerebral edema and epileptic seizures occurred. This is the first report of a woman with severe PMS and cerebral edema being treated successfully with an oral contraceptive containing drospirenone. PMID- 16373243 TI - Where are we with postmenopausal hormone therapy in 2005? AB - Recently, two large randomized placebo-controlled studies on long-term postmenopausal hormone replacement therapy, the Heart and Estrogen/progestin Replacement Study (HERS) and the Women's Health Initiative (WHI), have raise a lot of controversies, especially on the cardiovascular aspects. We briefly review these two trials and discuss what we know and do not know about postmenopausal hormone therapy, including the evidence on novel choices such as raloxifene and tibolone. PMID- 16373244 TI - The effect of red clover isoflavones on menopausal symptoms, lipids and vaginal cytology in menopausal women: a randomized, double-blind, placebo-controlled study. AB - BACKGROUND: The unexpected results of the Women's Health Initiative study have decreased the use of conventional hormone therapy (HT), changing physicians' and patients' attitudes towards HT and increasing their interest in alternative options. OBJECTIVE: The present study aimed to evaluate the effect of isoflavones contained in red clover extracts (Trifolium pratense) on menopausal symptoms, lipids and vaginal cytology in menopausal women. METHODS: Sixty postmenopausal women aged >40 years, non-users of HT, with Kupperman index score 15, were double blindly randomized to receive either a commercially available red clover isoflavone supplement (80 mg/day) or placebo for 90 days. Subsequently, after a 7 day washout period, subjects switched to receive the opposite treatment for a further 90 days. Kupperman index score was determined and fasting blood and vaginal cytologic sampling performed at baseline, 90 and 180 days. RESULTS: Fifty three women (88.3%) completed the trial. Mean age was 51.3 +/- 3.5 years, 69.7% of the women were aged 50 years or more. There was no significant effect on body mass index, weight or blood pressure after either treatment phase. Baseline Kupperman index score decreased significantly after each treatment phase, with the decrease more pronounced after the isoflavone phase (baseline: 27.2 +/- 7.7; after isoflavone: 5.9 +/- 3.9; after placebo: 20.9 +/- 5.3, p < 0.05). Red clover isoflavone supplementation significantly decreased the rate of menopausal symptoms and had a positive effect on vaginal cytology as expressed by improvement in karyopyknotic, cornification and basal cell maturation indices. Mean total cholesterol, low-density lipoprotein-cholesterol and triglyceride levels also decreased; however, only the latter was significantly lower compared with placebo. CONCLUSIONS: Compared with placebo, red clover isoflavone supplementation in postmenopausal women significantly decreased menopausal symptoms and had a positive effect on vaginal cytology and triglyceride levels. PMID- 16373245 TI - Efficacy and safety of 17alpha-hydroxyprogesterone caproate in hormone replacement therapy. AB - The aim of the present study was to evaluate the efficacy (in terms of induction of uterine bleeding) and safety (in terms of absence of endometrial hyperplasia) of 17alpha-hydroxyprogesterone caproate (17alpha-HPC) in a therapeutic regimen for hormonal replacement after menopause. Fifty postmenopausal patients received hormone replacement therapy (HRT) for 24 weeks. The treatment regimen consisted of standard estrogen replacement therapy at commonly prescribed doses for the prevention of climacteric symptoms and 341 mg of 17alpha-HPC every 30 days. Enrolled women were told to expect withdrawal bleeding 7-10 days after the administration of 17alpha-HPC. Forty-eight patients completed the trial. In 91.7% of cases, patients experienced the expected pattern, i.e., strict withdrawal bleeding exclusive of any other form of bleeding. Breakthrough bleeding and/or other forms of abnormal bleeding affected only four women. At the 6th month none of the endometrial samplings motivated by endometrial thickness >10 mm and evidence of heterogeneous echogenicity (two cases) was positive for carcinoma. No biopsies had to be performed at the end of the 12th month of treatment. No serious adverse effect where recorded during the study period. In conclusion, our data show the efficacy and safety of 17alpha-HPC in HRT. PMID- 16373246 TI - 3alpha-reduced neuroactive steroids and their precursors during pregnancy and the postpartum period. AB - Allopregnanolone (ALLO) and pregnanolone (PREG), the 3alpha-reduced metabolites of progesterone (PROG), are potent modulators of gamma-aminobutyric acid type A receptors that may function as endogenous anxiolytics. They are purported to be involved in the etiology or expression of clinical depression. In the present study we quantified ALLO and PREG, as well as PROG, 5alpha-dihydroprogesterone (5alpha-DHP), 5beta-dihydroprogesterone (5beta-DHP), epiallopregnanolone and pregnenolone (PREGNEN), in plasma from healthy women at five time points during pregnancy and the postpartum period. Analysis was by gas chromatography/electron capture - negative chemical ionization - mass spectrometry. Neuroactive steroids increased significantly from 10 to 36 weeks of pregnancy, except for 5beta-DHP and PREGNEN which did not change significantly. PROG was the most abundant steroid throughout pregnancy, followed by 5alpha-DHP and ALLO. Metabolite to precursor ratios differed depending on the enzyme and substrate: the turnover of PROG to 5alpha-DHP (catalyzed by 5alpha-reductase) was stable while the conversion of PROG to 5beta-DHP (catalyzed by 5beta-reductase) decreased later in pregnancy. 3alpha-Hydroxysteroid oxidoreductase-mediated turnover of 5alpha- and 5beta-DHP to their metabolites ALLO and PREG, respectively, rose during pregnancy, but the turnover of 5alpha-DHP to ALLO dropped at the late prenatal visit. At 6 weeks postpartum all steroids were significantly reduced compared with late prenatal values, with 5alpha-DHP being the most abundant postpartum steroid. These results provide the basis for further study of neuroactive steroids in psychiatric conditions of pregnancy and the postpartum period. PMID- 16373247 TI - Consequences of menopause in women with diabetes mellitus - a clinical problem. AB - Human life was prolonged by 30 years in the past century, with the result that about 40% of a woman's life falls within the postmenopausal period. The consequences, both early and remote, in the form of cardiovascular disease, osteoporosis and neoplastic disease are most pronounced in women suffering from one of the most common diseases, i.e., diabetes mellitus and the metabolic syndrome preceding it. These patients are problematic for physicians, and for this reason a study of diagnostic and therapeutic management was undertaken on the basis of our own experience as well as examination based on evidence-based medicine. Prior to making therapeutic decisions it is necessary to determine cardiovascular, thromboembolic and breast cancer risk factors. Hormonal therapy may be helpful in young postmenopausal women who are free of risk factors, and its composition and route of administration are significant considerations. Women with risk factors and who are more than 10 years after menopause should be administered alternative therapy depending on the diagnosed pathology. PMID- 16373248 TI - Cerebral artery hemodynamics in polycystic ovary syndrome. AB - OBJECTIVE: The aim of the present study was to investigate hemodynamic changes in the medial cerebral artery and also the internal carotid artery in young women with polycystic ovary syndrome (PCOS) and polycystic ovaries (PCO). METHODS: Twenty-eight patients with PCOS, 16 patients with PCO and 24 healthy control subjects were included in the study. Blood flow rate, pulsatility index and back pressure of both the medial cerebral artery and the internal carotid artery were determined by transcranial Doppler ultrasonography and the results compared between groups. RESULTS: There were no significant differences between the groups in bilateral medial cerebral artery and internal carotid artery blood flow rate, pulsatility index and back pressure. CONCLUSION: Our results do not indicate whether the risk of cerebrovascular events will increase for PCOS patients in middle and advanced age, but do show that changes in cerebral hemodynamics are not likely in PCOS at an early stage. PMID- 16373249 TI - Does plasma insulin level affect ovarian response to exogenous administration of follicle-stimulating hormone in women without polycystic ovary syndrome? AB - BACKGROUND: Raised insulin levels have been shown to contribute to ovarian overproduction of androgens. Hyperinsulinemia, usually associated with polycystic ovary syndrome (PCOS), brings about greater ovarian endocrine and morphological responses to ovulation induced by follicle-stimulating hormone (FSH). This indicates that elevated levels of insulin play a role in the endocrine and paracrine control of the ovaries. OBJECTIVE: The aim of the present study was to investigate whether basal insulin levels influence ovarian response to FSH in healthy women (non-PCOS) undergoing assisted reproduction by in vitro fertilization-embryo transfer (IVF-ET). METHODS: The study included 36 consecutive women, 27-45 years old, undergoing IVF-ET for tubal-factor or male factor infertility. Serum insulin levels were determined on the day of administration of gonadotropin-releasing hormone analog (GnRHa) and on the first day of FSH administration. RESULTS: Mean insulin levels were 6 +/- 3 and 7 +/- 3 microU/ml on the day of GnRHa and FSH administration, respectively. No correlations were found between basal insulin level, days of treatment, total FSH dose, estradiol level and the number of oocytes retrieved. CONCLUSIONS: The results of the present study show that normal levels of insulin do not seem to influence ovarian response to FSH in non-PCOS women. In all patients included in our study, serum insulin levels did not correlate with IVF stimulation data (days of stimulation, total FSH dose) nor with IVF-ET outcome. Thus the study demonstrates that, in patients who are not insulin-resistant, insulin does not correlate with ovarian response to FSH administration. PMID- 16373250 TI - Relationship of body fat distribution by waist circumference, dual-energy X-ray absorptiometry and ultrasonography to insulin resistance by homeostasis model assessment and lipid profile in obese and non-obese postmenopausal women. AB - OBJECTIVE: To correlate body fat distribution evaluated by waist circumference, dual-energy X-ray absorptiometry and ultrasonography to insulin resistance and lipid profile in obese and non-obese postmenopausal women. METHODS: We studied 40 obese and 47 non-obese postmenopausal women, assessing obesity by measuring waist circumference and fat tissue using dual-energy X-ray absorptiometry and ultrasonography, and examining their correlation with metabolic parameters: insulin resistance as determined by the homeostasis model assessment technique (HOMA-IR) and lipid profile including triglycerides (TG), total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), very-low-density lipoprotein, lipoprotein(a) (Lp(a)) and apoplipoprotein A-I (Apo A-I). RESULTS: There was no difference in lipid profile between the two groups. Insulin resistance was the metabolic disturbance of highest prevalence in the obese group, evaluated by HOMA-IR (obese: 3.38 +/- 2.2; non-obese: 1.20 +/- 0.7; p < 0.001). Obesity was not a confounding factor in linear regression analyses among HOMA-IR, HDL-C, TG, Lp(a), Apo A-I and the methods used to measure body fat distribution. Waist circumference was the method that best explained HOMA-IR (R(2) = 34.9%, p < 0.001) and TG concentration (R(2) = 10.9%, p = 0.002). HDL-C presented a positive association with subcutaneous fat evaluated by ultrasonography (R(2) = 12.5%, p < 0.001). Obesity was a confounding factor in multiple regression analyses between TC and LDL-C, when related to abdominal fat evaluated by ultrasonography, and resulted in a positive association among the obese and a negative association among the non-obese women. The sensibility of this method was related to the quantity of fat in the visceral region. CONCLUSIONS: Waist circumference showed the highest association with insulin resistance. Fat distribution evaluated by dual-energy X-ray absorptiometry and ultrasound was also associated with insulin resistance, but with lower intensity. The relationship of visceral fat distribution evaluated by ultrasound to TC cholesterol and LDL-C showed opposed results between obese and non-obese menopausal women. PMID- 16373251 TI - Environment and systemic lupus erythematosus: an overview. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease of unknown etiology that manifests as a pleomorphic systemic disease mainly affecting females. The variety of autoantibodies found in the serum of patients indicate that SLE is an autoimmune disease, but the mechanisms leading to the aberrant responses are not clearly understood although it is thought that a number of genetic and environmental factors may be involved. Environmental (or non-genetic) exposures could include infectious agents, chemicals or other compounds capable of modulating immune responses such as occupational/environmental pollutants or drugs, and behavioural factors such as smoking and diet. Environmental exposures may lead to the production of autoreactive T cells and autoantibodies, the stimulation of pro- and antiinflammatory cytokines, and target end-organ damage, but are not so convincing as agents causing SLE. Exposure to viruses increases antibody titres, but these may be the result of polyclonal B cell activation. The amount and timing of exposure to different environmental factors may play a significant and complex role in the pathogenesis of SLE and other autoimmune diseases. A better understanding of the etiopathogenetic mechanism of SLE is required in order to clarify the multiple interactions between environmental exposures and genetic factors. PMID- 16373252 TI - Infections and SLE. AB - Viral and bacterial infections may serve as an environmental trigger for the development or exacerbation of systemic lupus erythematosus (SLE) in the genetically predetermined individual. In addition, SLE patients are more prone to develop common (pneumonia, urinary tract infection, cellulitis, sepsis), chronic (tuberculosis), and opportunistic infections possibly due to inherit genetic and immunologic defects (complement deficiencies, mannose-binding lectin [MBL] polymorphisms, elevated Fcgamma III and GM-CSF levels, osteopontion polymorphism), but also due to the broad spectrum immunosuppressive agents that are part of therapy for severe manifestations of the disease. Hence, SLE patients are considered a high-risk population, where identification and treatment of chronic infections such as tuberculosis, hepatitis B or human immunodeficiency virus, are important prior to the institution of immunosuppression so as to prevent reactivation or exacerbation of the infection. Infections in SLE patients remain a source of morbidity and mortality. A caveat often encountered is to distinguish between a lupus flare and an acute infection; in such cases parameters including elevated CRP (and adhesion molecules) may aid in the diagnosis of infection. Recent research has provided convincing evidence that EBV infection may play a major role not only in molecular mimicry but also in aberrations of B cells and apoptosis leading to a state of perpetual heightened immune response in SLE. PMID- 16373253 TI - Human cytomegalovirus in patients with systemic lupus erythematosus. AB - The objective of this study was to determine the frequencies of human cytomegalovirus (HCMV) infection and HCMV genome copy number in blood of consecutive (treated from several months to several years) systemic lupus erythematosus (SLE) patients (22 women). The obtained results were compared to the healthy controls (15 women). All patients fulfilled at least four of the 1982 revised American rheumatism association (ARA) classification criteria for SLE. Our patients demonstrated three or four of the nine possible organ systems involved and most of them had mild SLE with SLE disease activity index (SLEDAI) score < 10 at time when blood samples were collected to detect HCMV. Quantitative analysis of HCMV genome was performed with aid of sequence analyzer ABI PRISM 7,700 Perkin Elmer. Primers and probe were constructed on the basis of IE4 region of HCMV genome. The viral load was expressed as log(10) of calculated HCMV genome copy number. Qualitative analysis revealed that 100% of our SLE patients were infected with HCMV, whereas in the control group only 73% of persons were HCMV positive. Statistically significant difference was demonstrated when the strength of the association between SLE or controls and infection of HCMV was calculated (estimated by Fisher's exact test, P value=0.02). Higher viral DNA copy number was observed in whole blood of SLE patients than in the control group (338.45+/- 221.76 and 229.00+/- 405.61 copies/ml respectively) but did not reach statistical significance level (95% confidence interval from 170.41 to 249.32, P=0.71). Furthermore percentage of patients with HCMV-DNA copy number >2.0 x 10(2) copies/ml was statistically significantly higher than this one in controls. The data show association between HCMV infection and SLE, which should be taken into account during the course of SLE. PMID- 16373254 TI - Pneumococcal vaccination of patients with systemic lupus erythematosus: effects on generation of autoantibodies. AB - OBJECTIVES: To assess the effect of vaccination against streptococcus pneumoniae on the generation of autoantibodies in patients with SLE. MATERIALS AND METHODS: Twenty-four consecutive patients with SLE were vaccinated against streptococcus pneumoniae. Assessment was performed the day of vaccination and 2 months later and included evaluation of disease activity using the SLEDAI, serum levels of ESR, CRP, C3 and C4. The sera of the patients were tested by ELISA for anti dsDNA, anticardiolipin (IgG and IgM), anti-Sm, anti-nRNP, anti-Ro/SSA, and anti La/SSB. RESULTS: The mean age at enrollment into the study was 39, mean disease duration 6.9 years. The SLEDAI score (mean +/- SD) was 4.41 +/- 2.92 at the time of vaccination and 4.47 +/- 3.11, 2 months apart. At the time of vaccination, 10 patients had anti-dsDNA, 2 patients had anti-Sm, 5 had anti-nRNP, and 9 had anti Ro/SSA, 4 had anti-La/SSB, 4 had anticardiolipin IgG and IgM. Two months after vaccination, no change was observed in the proportion of patients with anti-Sm, anti-dsDNA, anti-RNP, anti-Ro/SSA and anticardiolipin IgM. A single patient developed anticardiolipin IgG and another one turned anti-RNP negative. CONCLUSIONS: Vaccination against streptococcus pneumoniae did not trigger the generation of autoantibodies and confirms the clinical safety of this vaccine in SLE patients. PMID- 16373255 TI - Occupational exposures and risk of systemic lupus erythematosus. AB - This review summarizes the growing body of epidemiologic and experimental research pertaining to the relationship between SLE and occupational exposures, such as crystalline silica, solvents, and pesticides. Epidemiologic studies, using different designs in different settings, have demonstrated moderate to strong associations between occupational silica exposure and SLE. Recent experimental studies of silica in lupus-prone mice provide support for the idea that, in addition to its known adjuvant effect, silica exposure increases the generation of apoptotic material, an important source of self-antigen. Despite compelling experimental studies of the organic solvent trichloroethylene (TCE) in lupus-prone mice, there is little evidence of an overall association of SLE and occupational exposure to a broad classification of solvents in humans. However, there is a lack of data on SLE in occupational cohorts with exposures to TCE or other specific solvents. One epidemiologic study reported an association of pesticide mixing and SLE, while a recent experimental study reported accelerated disease in pesticide-treated lupus-prone mice. Other occupational exposures worth investigating include asbestos, metals, and UV radiation. Attention should also be given to the role of gene-environment interactions, which may require large, multi-site studies that collect both genetic material and occupational exposure data. The quality of exposure assessment is an important consideration in designing and evaluating these studies. The use of pre-clinical endpoints (e.g. high-titer autoantibodies) in occupational cohorts with well-characterized exposure histories may reveal occupational risk factors for autoimmunity, and may also provide baseline data for studies of determinants of progression to SLE. PMID- 16373256 TI - Drug-induced lupus erythematosus. AB - Drug-induced lupus is a syndrome which share symptoms and laboratory characteristics with idiopathic systemic lupus erythematosus (SLE). The terms drug-induced lupus (DIL) and drug-induced lupus erythematosus (DILE) are preferred, but other ones are also used-drug-related lupus, lupus-like syndrome and lupus erythematosus medicamentosus. The first case of DILE was reported in 1945 and associated with sulfadiazine. In 1953, it was reported that DILE was related to the use of hydralazine. More than 80 drugs have been associated with DILE. The average age of patients with DILE is nearly twice that of patients with idiopathic SLE. Approximately half the patients with drug-induced SLE are women, compared with 90% of patients with idiopathic SLE. Similarly to idiopathic lupus, DILE can be divided into systemic, sub-acute cutaneous and chronic cutaneous lupus. The syndrome is characterised by arthralgia, myalgia, pleurisy, rash and fever in association with antinuclear antibodies in the serum. The clinical and laboratory manifestations of drug-induced SLE are similar to those of idiopathic SLE, but central nervous system and renal involvement are rare in DILE. Recognition of DILE is important because it usually reverts within a few weeks after stopping the drug. This review discusses the general issues in DILE, such as pathogenic mechanisms, clinical forms and diagnostic criteria, and provides more detailed information for some of the most recent implicated drugs: minocycline, statins, anti-TNF-alpha agents. PMID- 16373257 TI - Photosensitivity in lupus erythematosus. AB - Lupus erythematosus (LE) is an autoimmune disease which can be triggered by environmental factors such as solar irradiation. It has long been observed that especially ultraviolet (UV) exposure can induce and exacerbate skin lesions in patients with this disease. However, despite the frequency of photosensitivity in LE, the mechanisms by which UV irradiation activates autoimmune responses is only now becoming increasingly unfolded by advanced molecular and cellular biological investigations. Phototesting, according to a standardized protocol with UVA and UVB irradiation has proven to be a valid model to study photosensitivity in various subtypes of LE and to evaluate the underlying pathomechanisms of this disease. Detailed analysis of the molecular events that govern lesion formation in experimentally photoprovoced LE showed increased accumulation of apoptotic keratinocytes and impaired expression of the inducible nitric oxide synthase (iNOS). In the near future, gene expression profiling and proteomics will further increase our knowledge on the complexity of the "UV response" in LE. This review summarizes the current understanding of the clinical and molecular mechanisms that initiate photosensitivity in this disease. PMID- 16373258 TI - Effects of dehydroepiandrosterone supplement on health-related quality of life in glucocorticoid treated female patients with systemic lupus erythematosus. AB - The objective of this study was to evaluate the efficacy of low dose dehydroepiandrosterone (DHEA) on health-related quality of life (HRQOL) in glucocorticoid treated female patients with systemic lupus erythematosus (SLE). Forty one women ( >or= 5 mg prednisolone/day) were included in a double-blind, randomized, placebo-controlled study for 6 months where DHEA was given at 30 mg/20 mg ( or= 46 years) daily, or placebo, followed by 6 months open DHEA treatment to all patients. HRQOL was assessed at baseline, 6 and 12 months, using four validated questionnaires and the patients' partners completed a questionnaire assessing mood and behaviour at 6 months. DHEA treatment increased serum levels of sulphated DHEA from subnormal to normal. The DHEA group improved in SF-36 "role emotional" and HSCL-56 total score (both p<0.05). During open DHEA treatment, the former placebo group improved in SF-36 "mental health" (p<0.05) with a tendency for improvement in HSCL-56 total score (p=0.10). Both groups improved in McCoy's Sex Scale during active treatment (p<0.05). DHEA replacement decreased high-density lipoprotein (HDL) cholesterol and increased insulin-like growth factor I (IGF-I) and haematocrit. There were no effects on bone density or disease activity and no serious adverse events. Side effects were mild. We conclude that low dose DHEA treatment improves HRQOL with regard to mental well being and sexuality and can be offered to women with SLE where mental distress and/or impaired sexuality constitutes a problem. PMID- 16373259 TI - Cigarette smoking and systemic lupus erythematosus: a smoking gun? AB - Cigarette smoking is one of many environmental exposures, including infectious agents, silica exposure, hormonal and dietary factors, hypothesized to be linked to the development of SLE. Cigarette smoke contains hundreds of potentially toxic components, including tars, nicotine, carbon monoxide, polycyclic aromatic hydrocarbons among others. Whether cigarette smoking increases the risk of developing SLE remains controversial. To date, three case-control studies have reported significantly increased odds ratios for the development of SLE in smokers, while six others have not found a clear association. A meta-analysis statistically combining the effect estimates from the available studies, revealed a modestly increased risk posed by current smoking (RR 1.5 [95% CI 1.09, 2.08]), but no increased risk associated with past smoking. The strengths and weaknesses of these data and potential implications are discussed. PMID- 16373260 TI - Positive and negative effects of thalidomide on refractory cutaneous lupus erythematosus. AB - BACKGROUND: Thalidomide is used in cutaneous lupus erythematosus (CLE) refractory to conventional therapies. Peripheral neuropathy (PN) is the most severe side effect, but the incidence of PN and its relation to thalidomide dose are still unclear. OBJECTIVE: To prospectively evaluate the efficacy as well as the occurrence of PN in CLE patients treated with thalidomide, and to assess whether PN, when occurs, correlates with thalidomide dose and/or length of treatment. METHODS: Fourteen female patients with CLE in low-dose thalidomide therapy were followed for up to 24 months. Prior to, and regularly during treatment patients underwent rheumatological, dermatological, neurological and electrophysiological evaluations. A decline in sural SNAP of 50% or more from baseline value was considered as criterion of sensory axonal PN. RESULTS: All patients showed a dramatic improvement of skin manifestations. Ten patients (71.4%) developed a sensory axonal PN. The median time free from this complication was 14 months. No correlations were found between age of the patients nor thalidomide cumulative dose and occurrence of PN (Mann-Whitney U Test; p>0.16). Other adverse effects were: tremor, paresthesias, somnolence, amenhorrea, constipation and thoracic pain. CONCLUSIONS: Low does thalidomide is efficacious in treating CLE, but PN is a common complication whose occurrence does not seem to correlate with total thalidomide dose, whereas with the duration of therapy. A closer electrophysiological follow-up is therefore recommended in the long-term treatment. PMID- 16373261 TI - MRI-guided myocardial cell therapy. PMID- 16373262 TI - Drug-eluting versus third-generation bare metal stents: the US strategy. AB - Drug-eluting stents have rapidly come to dominate the field of coronary intervention, constituting 85% of the US market just one year after introduction. The current utilization of bare metal stents in coronary intervention in the US is quite limited. This article will review technological advances in third generation bare metal stents, which optimize procedural performance by means of improved flexibility, lower crossing profile, and thinner struts while maintaining fluoroscopic visibility and may portend a continued role for bare metal stents in some settings. This article will review data pertaining to outcomes with new generation bare metal stents and discuss current options for utilization of drug eluting stents versus bare metal stents in the US. PMID- 16373263 TI - Embolic protection: limitations of current technology and novel concepts. AB - Distal embolic event is one of the major limitations of coronary and non-coronary vascular interventions. Balloon and filter-based Embolic Protection Devices (EPDs) are a new class of interventional devices, used to prevent consequential morbidity and mortality of the distal embolic events. Data from first generation EPD supply proof of concept and show approximately 40% reduction in mortality and morbidity, when EPDs are used during saphenous vein grafts (SVGs) interventions. Current limitations of all first generation EPD technology taper their penetration. With breakthroughs in embolic protection technology, it is estimated that, in the near future, EPDs will be used with stenting in all high-risk lesions (SVGs, carotid arteries and acute coronary syndromes), become the standard of care and even be used in low risk cases. PMID- 16373264 TI - The ACIST power injection system reduces the amount of contrast media delivered to the patient, as well as fluoroscopy time, during diagnostic and interventional cardiac procedures. AB - The ACIST injection system is an automatic power injection device that allows for online control of injection rate and volume of contrast. Limited data is available whether this technology allows reducing use of contrast and fluoroscopy time. Accordingly, we compared the use of this system to manual injection among 450 consecutive patients who underwent diagnostic coronary angiography and/or angioplasty who were randomly assigned to either manual contrast injection (control; n=198) or to the ACIST system (study group; n=252). The amount of contrast, fluoroscopy and total procedural times were recorded for each patient. In the diagnostic group, the mean total amount of contrast (including wasted) was reduced by 63% when the ACIST was used compared to control (100+/-42 ml versus 163+/-56 ml; P<0.001, respectively). When only the net amount of contrast delivered to the patient was considered, the differences were smaller (20%, P=0.004). During angioplasty, the amount of contrast was also lower in the ACIST group (206+/-65 versus 230+/-69, P=0.008), whereas no difference were noted in net amount of contrast. Fluoroscopy time was significantly shorter in the ACIST group compared to control both during diagnostic catheterization (4.7+/-3.5 min versus 6.3+/-5.5 min, respectively; P=0.014), and angioplasty (16.7+/-9.1 min versus 19.6+/-12.4 min, respectively; P=0.05). Routine utilization of the ACIST system during diagnostic and interventional procedure significantly reduced the total amount of contrast media used and fluoroscopy time. PMID- 16373265 TI - Drug-eluting stents for the treatment of in-stent restenosis--'real world' double centre experience in consecutive patients. AB - OBJECTIVES: We evaluated clinical outcome of consecutive patients with in stent restenosis (ISR) treated with drug-eluting stents (DES) at two intervention centres from April 2002 to April 2004, reflecting 'real world' practice. BACKGROUND: ISR is the major limitation to successful long-term outcome after implantation of bare metal stents during percutaneous coronary intervention (PCI). The optimal strategy for the treatment of ISR has yet to be determined. METHODS: 121 consecutive patients with significant ISR were treated with DES. Sirolimus DES were used in 60 patients and paclitaxel DES in the remainder. All patients were followed up to evaluate the incidence of major adverse cardiac events (MACE), angina class and clinically driven angiography. Data were collected between 7 and 27 months after the procedure (mean follow up of 16.5 months). RESULTS: Overall MACE rate at 16.5 months was 13.2% (16 patients) including 4 deaths (3%). Fifteen (12.3%) patients underwent clinically driven angiography. Eight patients (6.6%) developed ISR within the treated segment, of whom, four underwent further PCI and 4 CABG. Mean Canadian angina class decreased from 2.46+/-0.7 pre-procedure to 0.69+/-0.6 at follow up. All patients achieved an improvement in angina, with 59% being rendered angina free and 87% free of MACE. There were no differences in clinical outcome in those who received a paclitaxel and sirolimus DES. CONCLUSIONS: The use of DES implantation for the treatment of ISR is safe, effective and associated with low recurrence rates in a 'real world' large cohort of patients with a complex mix of anatomical and clinical factors. PMID- 16373266 TI - Characteristics and clinical outcomes of patients with cardiogenic shock complicating acute myocardial infarction treated by emergent coronary angioplasty. AB - BACKGROUND: Cardiogenic shock (CS) is a dreadful complication of acute myocardial infarction (AMI) associated with a poor prognosis. Percutaneous coronary intervention (PCI) is widely recommended by current treatment guidelines. AIM: To evaluate the in-hospital and 30-day mortality rate and to determine independent predictors of mortality in a cohort of unselected consecutive patients with CS. METHODS AND RESULTS: Rabin Medical Center cardiac catheterization laboratory database was analyzed between 1/2000 and 8/2003. Fifty of the 472 patients (10.6%) treated using emergent PCI for AMI had cardiogenic shock on presentation. Patients with cardiogenic shock were older, more likely to be female and with higher frequency of co-morbidities. The time from symptom onset until seeking medical treatment was longer in cardiogenic shock patients. In-hospital mortality rate was 48.0% in the cardiogenic shock group as compared to 3.3% in the non cardiogenic shock group (P<0.0001). In patients with shock, total mortality after 30 days was 52% (26/50). Most of these patients (25/26) died within 48 hours following admission because of refractory cardiogenic shock. A multivariate analysis adjusted for baseline differences showed that age 75 years (odds ratio [OR]: 11; 95% confidence interval [CI]: 1.0-1.24, P=0.05), and the use of GP 2b/3a antagonist (OR: 0.97; 95% CI: 0.95-1.0, P=0.05), were independent predictors of all cause mortality at 30 days. CONCLUSION: Cardiogenic shock remains an important cause of mortality in AMI. Younger age and the use of GP 2b/3a antagonists during primary PCI for cardiogenic shock patients seems to be associated with better clinical outcomes. PMID- 16373267 TI - Management of calcified lesions in 2004. AB - Calcified lesions are encountered with increasing frequency in the catheterization laboratory. Percutaneous coronary interventions of calcified lesions are associated with a higher complication rate than percutaneous intervention on non-calcified lesions. The review focuses on current management strategies in the drug eluting stent era of this complex lesion subset. Evidence based approaches to treat calcified coronary lesions with balloon based and atherectomy devices as adjunctive modalities in the drug eluting stent era are discussed in this article. PMID- 16373270 TI - Endogenous opioids and liver disease. PMID- 16373271 TI - Dietary supplementation of carbonate promotes spontaneous tumorigenesis in a rat gastric stump model. AB - OBJECTIVE: Food supplements are known to affect the development of gastric adenocarcinoma. In this study, an animal model of gastric resection was used to investigate the effects of calcium carbonate on spontaneous development of gastric adenocarcinoma. MATERIAL AND METHODS: Ninety-two Wistar rats with gastric resections (performed to induce spontaneous gastric cancer) and 60 without resections (controls) were used to analyse the carcinogenic potential of different ion supplements in food. RESULTS: Among the resected rats, cancer developed in 3 out of 18 (17%, NS) given NaCl but in 11 out of 18 (61%, p<0.01) exposed to calcium carbonate. No tumours were found in the unresected (unoperated) animals. These findings were further analysed by separately investigating the effects of calcium and carbonate ions on tumorigenesis in the gastric stump model. Cancer developed in one of 26 (4%) resected animals given a diet supplemented with CaHPO(4), which was lower than the rate observed in the resected control group fed a normal diet, although this difference was not statistically significant. However, tumour development increased significantly in the resected animals given a diet supplemented with NaHCO(3) (tumours in 13 out of 24 rats, 54%; p<0.01). CONCLUSIONS: The present results reveal a significant role for carbonate in the induction of gastric carcinoma in the rat. The relevance of this finding is underlined by the fact that carbonate is a major constituent of intestinal reflux into the stomach, and that such reflux is considered to be one of the major causes of gastric cancer. PMID- 16373272 TI - Cost-effectiveness of Helicobacter pylori "test and treat" for patients with typical reflux symptoms in a population with a high prevalence of H. pylori infection: a Markov model analysis. AB - OBJECTIVE: Patients with typical reflux symptoms may have underlying peptic ulcer disease (PUD) in a population with a high prevalence of Helicobacter pylori infection. In the present study we sought to compare the cost-effectiveness of the H. pylori "test and treat" strategy with that of empirical proton-pump inhibitor (PPI) therapy and endoscopy in patients with typical reflux and a high prevalence of H. pylori infection. MATERIAL AND METHODS: A Markov model was designed to compare the outcomes of three treatment strategies over 12 months among a hypothetical cohort of patients presenting with typical reflux symptoms. There were four exclusive underlying H. pylori- and PUD-related comorbidities in the hypothetical cohort: (1) H. pylori-related PUD, (2) H. pylori infection without PUD, (3) PUD without H. pylori infection and (4) absence of both PUD and H. pylori infection. The transition probabilities and resource utilization were derived from the literature. Percentage of PUD patients treated, total number of symptom-free patient-years gained and total direct medical cost were estimated. RESULTS: By comparing each strategy individually with the no therapy strategy, it was found that the incremental costs per ulcer treated for H. pylori "test and treat", endoscopy and empirical PPI therapy were USD 1778, USD 1797 and USD 2158, respectively. The results of the model were sensitive to the prevalence of H. pylori infection. CONCLUSIONS: Both the H. pylori "test and treat" and initial endoscopic strategies were shown to be more cost-effective than empirical PPI therapy for treating undiagnosed PUD among patients presenting with typical reflux symptoms. The H. pylori "test and treat" strategy appeared to be only slightly more cost-effective than initial endoscopy in a population with a high prevalence of H. pylori infection. PMID- 16373273 TI - Reverse gastric pacing reduces food intake without inducing symptoms in dogs. AB - OBJECTIVE: To investigate the therapeutic potential of reverse gastric pacing (RGP) for obesity by studying the effects of RGP on food intake and symptoms in dogs. MATERIAL AND METHODS: The study was performed in 9 dogs in 3 sessions (control, strong RGP and moderate RGP). Gastric myoelectrical activity and food intake were measured in each session. RGP was performed using serosal electrodes implanted in the distal stomach at the physiological frequency of the intrinsic gastric myoelectrical activity with a pulse width of 550-950 ms. The amplitude of the stimulus was set at a level maximally tolerable by the animals in the strong RGP session and 50% of the maximum level in the moderate session. RESULTS: 1) Compared with the control, strong RGP (p<0.001) and moderate RGP (p<0.01) significantly reduced the amount of food intake by 62.9% and 31.7%, respectively (p<0.05, ANOVA). 2) Whereas strong RGP induced significant symptoms, moderate RGP did not induce any significant symptoms in comparison with the control session. 3) The regularity and coupling of gastric myoelectrical activity were significantly impaired with both strong RGP and moderate RGP in the fasting state. CONCLUSIONS: RGP impairs intrinsic gastric myoelectrical activity and substantially and acutely reduces food intake. The reduced food intake and freedom from symptoms resulting from moderate RGP are indicative of the therapeutic potential of RGP in obesity. PMID- 16373274 TI - Low lactase activity in a small-bowel biopsy specimen: should dietary lactose intake be restricted in children with small intestinal mucosal damage? AB - OBJECTIVE: Small intestinal mucosal damage can result in decreased lactase activity (LA). When LA is low in a small-bowel biopsy (SBB) specimen, a reduction of dietary lactose intake is usually advised. This is often done by reducing dietary dairy products, which also reduces the intake of calcium, protein and vitamins. Since intestinal damage can have a patchy character and LA varies along the horizontal axis of the small intestine, the relevance of SBB measurement for intestinal LA could be questioned. We compared LA in the SBB with the in vivo capacity to digest lactose using the Lactose Digestion Index (LDI). MATERIAL AND METHODS: LA was measured in 18 children aged 0.8-10.9 years (mean 3.9, SD 2.4) undergoing SBB for various indications. In all children the LDI was determined using the (13)C-lactose/(2)H-glucose test. RESULTS: In 9/18 biopsy specimens LA was low (<10 U/g protein). LDI was normal in 14/18 patients. In 8 out of 9 patients with normal lactase activity, LDI was also normal, while in 6 out of 9 patients LDI was normal despite low LA in the biopsy. In patients with normal LDI, histology was normal in 6/14, in 4/14 mild histological changes (Marsh II) were seen and in 4 patients histological damage was severe (grade III). CONCLUSIONS: In children with small-bowel mucosal damage, lactose digestive capacity can remain high despite low LA and histological changes in an SBB. Extrapolation of LA in SBB specimens to overall lactose digestive capacity may not be reliable. The advice concerning the restriction of intake of dairy products cannot be based on the data of the SBB only. PMID- 16373275 TI - Coeliac children on a gluten-free diet with or without oats display equal anti avenin antibody titres. AB - OBJECTIVE: Recent studies report negligible toxicity of oats in the majority of coeliac disease (CD) patients. It has previously been shown that children with untreated CD have circulating antibodies to oats avenin. In this study we performed serial assessments of anti-avenin antibodies in children under investigation for CD on a gluten-free diet with or without oats. MATERIAL AND METHODS: The study involved 116 children, randomized to a standard gluten-free diet or a gluten-free diet supplemented with oats. Sera were obtained from 86 children, 48 in the standard gluten-free group and 38 in the gluten-free oats group, of which 33 consumed at least 10 g of oats daily. IgA and IgG anti-avenin antibodies were monitored at 0, 3, 6 and 12 months. Nitric oxide metabolites were measured in 7 patients, with deviating antibody results. RESULTS: There was a significant decrease in anti-avenin antibodies in both groups at the end as compared to the beginning of the study, (p<0.001), but no difference was found between the two groups. IgA titres already declined after 3 months. IgG titres, although significantly decreased, remained high in the majority of patients in both groups. Nitric oxide levels were high in four of the analysed samples. CONCLUSIONS: Oats per se, do not seem to produce a humoral immune reaction in children with CD when given in an otherwise gluten-free diet, indicating that the reaction requires gluten challenge. Anti-avenin antibodies were equal in the two study groups, and these findings strengthen the clinical impression that oats can be tolerated by the majority of patients with CD. PMID- 16373276 TI - Lack of association between IBD5 and Crohn's disease in Japanese patients demonstrates population-specific differences in inflammatory bowel disease. AB - OBJECTIVE: Population-specific differences in the genetic susceptibility to inflammatory bowel disease (IBD) are indicated by the fact that Crohn's disease (CD) in Japanese patients does not have any of the common CARD15 variants that are associated with CD in Caucasians. Recently, the disease-causing mutation in the IBD5 haplotype was identified. The TC haplotype, composed of L503F in SLC22A4 and -207G/C in SLC22A5 promoters, was reported to alter the function of the organic cation transporter and to be associated with CD in Caucasians. To determine whether the TC haplotype is also associated with IBD in a Japanese population, we genotyped L503F and -207G/C variants in Japanese subjects. Furthermore, we also performed a case-control association study with all representative single nucleotide polymorphisms (SNPs) in IBD5 using previous information of linkage disequilibrium extension reported in Japanese patients to determine whether there were variants in IBD5 specifically associated with IBD in Japanese patients. MATERIAL AND METHODS: A total of 758 Japanese individuals, 241 patients with CD, 247 patients with ulcerative colitis (UC) and 270 healthy controls, were analyzed in this study. Genotyping for SNPs was determined by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: We found L503F and -207G/C to be very rare (<1% frequency) in CD, UC and HC in the Japanese population. Furthermore, we also found that none of the representative SNPs in IBD5 was associated with CD or UC in the Japanese subjects. CONCLUSIONS: In contrast to Caucasians, IBD5 is not a major component of the susceptibility to IBD in the Japanese population. PMID- 16373277 TI - Detection of inflammatory markers in stools from patients with irritable bowel syndrome and collagenous colitis. AB - OBJECTIVE: Irritable bowel syndrome (IBS) and collagenous colitis (CC) share chronically recurring symptoms of altered bowel habits associated with abdominal pain or discomfort. The aims of the present study were to investigate whether inflammatory markers could be detected in faeces from patients with IBS and CC, and to elucidate whether such analyses could be used as non-invasive tools to distinguish between these disorders. MATERIAL AND METHODS: Stool samples were obtained from 18 patients with CC, 46 patients with IBS and 20 healthy controls (HC). Eosinophil protein X (EPX), myeloperoxidase (MPO), tryptase, interleukin-1 beta (IL-1beta) and tumour necrosis factor alpha (TNFalpha) were measured in supernatants from processed faeces using immunoassays. RESULTS: EPX levels were enhanced in faeces from CC patients (median 3.8 microg/g (0.47-16.2)) compared to patients with IBS (0.44 microg/g (0.25-1.8)), p<0.001, and HC (0.46 microg/g (0.21-1.3)), p<0.001. In addition, MPO was increased in CC patients (11.7 microg/g (2.0-124)) compared to IBS patients (1.7 microg/g (0.81-5.2)), p<0.01, and HC (2.5 microg/g (1.1-6.3)), p<0.05. Tryptase was found in 9/18 patients with CC, 6/46 with IBS and 1/19 HC. IL-1beta was only enhanced in 2/11 CC patients and TNFalpha was not detected in any sample. CONCLUSIONS: Increased levels of EPX, MPO and tryptase were observed in stools from collagenous colitis patients, whereas the levels in IBS patients did not differ from healthy controls. Our data suggest that faecal markers could be used as part of the clinical work-up to determine which patients should be biopsied and evaluated for collagenous colitis. PMID- 16373278 TI - Expression and activation of MMP -2, -3, -9, -14 are induced in rat colon after abdominal X-irradiation. AB - OBJECTIVE: Colonic response to single-dose irradiation is characterized by epithelial denudation followed by restitution. Extracellular matrix (ECM) remodeling is involved in both of these phases. The aim of this study was to characterize the contribution of matrix metalloproteinases (MMPs) and of their stimulatory and inhibitory pathways in radiation-induced ecm remodeling in colonic tissue. MATERIAL AND METHODS: Rats were irradiated with single-dose 10 Gy X-rays to the abdomen. Activity, localization, and mRNA levels of MMPs and molecules involved in their activation and inhibition (plasmin/plasminogen; TIMPs), of inflammatory mediators (IL-1beta, TNF-alpha) in the distal colon, 1, 3, and 7 days after irradiation were analyzed using a combination of approaches including zymography, immunohistochemistry, and real-time reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: The main finding of this study is that radiation-induced alteration of the mucosal structure is concomitant with local increased expression and activation of MMP subtypes involved in basement membrane degradation (MMP-2, -3, and -9). We investigated MMP-2 activation pathways and found an early increase in mRNA levels of soluble inflammatory mediators (TNF-alpha and IL-1beta). Furthermore, transcription and activity of MMP-2 activating molecules, such as MMP-14, and molecules involved in the plasminogen/plasmin system were found to increase during the denudation phase. Interestingly, induction of MMP inhibitors TIMP-1 and PAI-1 was observed during the restitution phase. MMP inhibitors may be able to stop acute wound healing response by inhibiting ECM degradation. CONCLUSIONS: This study brings new insights into ECM remodeling in the colon after exposure to ionizing radiation and highlights the role of MMP subtypes specialized in basement membrane degradation. PMID- 16373279 TI - Estimated prevalence of hereditary cancers and the need for surveillance in a Norwegian county, Telemark. AB - OBJECTIVE: The aim of the study was to estimate the prevalence of hereditary cancers and the need for surveillance in Telemark county, Norway. MATERIAL AND METHODS: All persons attending the Norwegian Colorectal Cancer Prevention (NORCCAP) trial in Telemark were interviewed about cases of cancer in the family. Diagnoses were verified, pedigrees constructed and families classified according to preset criteria aiming at identifying hereditary cancer. Mutation analyses were performed in kindreds at risk for breast cancers when possible. Immunohistochemistry of tumors in assumed inherited colorectal cancer families was undertaken. RESULTS: The screening examination was attended by 7,224 persons among whom 2,866 had cancer in the family. Of these, 2,479 had no suspicion of any known inherited cancer syndrome. Family information questionnaires were mailed to 387 persons and returned by 191. Sixty-four of these 191 met the clinical criteria for familial cancer by family history after verification of diagnoses. Observed prevalences for being at risk for hereditary breast and breast-ovarian cancer (HBOC) or hereditary non-polyposis colorectal cancer (HNPCC) were 2.8 per thousand and 0.77 per thousand, respectively. CONCLUSIONS: The number of colonoscopies and mammograms obtained per year serving those who needed them was limited and reduced by clinical genetic work-up from 2,866 with a family history of cancer to 64 proven cases. Continued surveillance of an unnecessarily high number leads to unjustified cancer worry, is costly and uses up health-care facilities. Genetic work-up is a one-time job that reduces input numbers to surveillance programs, provides a starting-point for mutation testing and is economically cost beneficial if inherited cancers are prevented or cured by the health-care programs offered. PMID- 16373280 TI - Acetaldehyde inhibits the formation of retinoic acid from retinal in the rat esophagus. AB - OBJECTIVE: It has already been demonstrated that the rat esophagus produces retinoic acid from retinol. In this study, this process is further characterized and the effect of acetaldehyde examined to elucidate the possible mechanisms behind the epidemiological evidence that the incidence of esophageal cancer is higher in alcoholics. MATERIAL AND METHODS: Rat esophageal samples were incubated with all-trans retinal and newly formed all-trans retinoic acid (ATRA) was quantified using high-performance liquid chromatography (HPLC). Furthermore, beta nicotinamide adenine dinucleotide (NAD)-dependent acetaldehyde oxidation by the rat esophagus was examined by tracing NAD reduction using a spectrophotometer. RESULTS: Rat esophageal samples produced ATRA from all-trans retinal in a NAD dependent manner and the potential was significantly attenuated by phenetyl isothiocynate, an ALDH inhibitor, or acetaldehyde depending on the concentration used. Rat esophageal samples also oxidized acetaldehyde of various concentrations NAD dependently. The ATRA formation potential that was temporarily inhibited by acetaldehyde was recovered to the control level by dialysis when the specimen was incubated with up to 50 microM of acetaldehyde. CONCLUSIONS: The rat esophagus produces retinoic acid from retinal. An ALDH isoform(s) is responsible for this process and physiological concentration of acetaldehyde hampers the process, probably in a competitive manner. Since the disturbance of retinoic acid supply has been implicated in carcinogenicity, this finding may, at least in part, explain the high incidence of esophageal cancer in alcoholics, especially in those with inactive ALDH 2 whose blood acetaldehyde levels become higher than those with active ALDH 2. PMID- 16373281 TI - Meal-induced changes in splanchnic blood flow and oxygen uptake in middle-aged healthy humans. AB - OBJECTIVE: For decades, the determination of changes in splanchnic blood flow and oxygen uptake after a meal has been used in the management of patients with suspected chronic intestinal ischaemia. However, little is known about the normal meal-induced responses. The aim of the present study was therefore to measure the splanchnic blood flow and oxygen uptake before and after a standardized meal in a group of middle-aged normal volunteers. MATERIAL AND METHODS: Splanchnic blood flow and oxygen uptake were determined at baseline and after a 3600-kJ mixed meal in 8 healthy women (50-70 years) and 10 healthy men (52-76 years). Splanchnic blood flow was measured during hepatic vein catheterization by indirect Fick principle with indocyanine green as the indicator. Splanchnic oxygen uptake was calculated from splanchnic blood flow and the arteriovenous oxygen difference. RESULTS: The meal induced a significant peak increase in splanchnic blood flow of 0.60 (0.26-1.07) l x min(-1) (mean, range) from a baseline level of 1.05 (0.66 1.33) l x min(-1). Splanchnic oxygen uptake showed a significant peak increase of 1.40 (0.44-4.13) mmol x min(-1) from a baseline level of 2.18 (1.41-3.31) mmol x min(-1). A close association was found between the meal-induced peak increases in splanchnic blood flow and oxygen uptake, but the variables were not related to gender or body surface area of the subjects. CONCLUSIONS: A 3,600-kJ mixed meal induces a significant increase in splanchnic blood flow and oxygen uptake in middle-aged healthy humans. Our data may be relevant for the evaluation of corresponding data from patients with suspected chronic intestinal ischaemia. PMID- 16373282 TI - Pain attacks in non-complicated and complicated gallstone disease have a characteristic pattern and are accompanied by dyspepsia in most patients: the results of a prospective study. AB - OBJECTIVE: The cardinal indication for surgical treatment of gallstones is pain attacks. However, following cholecystectomy, 20% of patients remain symptomatic. It is unclear to what extent post-cholecystectomy symptoms can be ascribed to persistence of preoperative symptoms or to new pathology. The pain and digestive pattern in gallstone patients has not been defined in a recent setting with ultrasonography as the diagnostic method. The aim of this study was to characterize a pain pattern that is typical for gallstone disease and to describe the extent of associated dyspepsia. MATERIAL AND METHODS: A total of 220 patients with symptomatic gallstone disease including complicated disease (acute cholecystitis and common bile duct stones) were interviewed using detailed questionnaires to disclose pain patterns and symptoms of indigestion. RESULTS: All patients had pain in the right upper quadrant (RUQ) including the upper midline epigastrium. The pain was localized to the right subcostal area in 20% and to the upper epigastrium in 14%, and in the rest (66%) it was more evenly distributed. An area of maximal pain could be defined in 90%. Maximal pain was located under the costal arch in 51% of patients and in the epigastrium in 41%, but in 3% behind the sternum and in 5% in the back. The pain was referred to the back in 63% of the patients. The mean visual analogue scale (VAS) score was very high: 90 mm on a 0-100 scale. A pattern of incipient or low-grade warning pain with a subsequent relatively steady state until subsiding in the same fashion was present in 90% of the patients. An urge to walk around was experienced by 71%. Pain attacks usually occurred in the late evening or at night (77%), with 85% of the attacks lasting for more than one hour and almost never less than half an hour. Sixty-six percent of the patients were intolerant to at least one kind of food, but only 48% to fatty foods. Symptoms of functional indigestion (gastroesophageal reflux, dyspepsia or irritable bowel symptoms) were seen in the vast majority in association with attacks. CONCLUSIONS: Gallstone-associated pain follows a certain pattern in the majority of patients. The pain is located in a defined area with a point of maximum intensity, is usually referred, and occurs mainly at night with duration of more than one hour. The majority of patients experience functional indigestion, mainly of the reflux type or dyspepsia. PMID- 16373283 TI - Serum osteopontin levels in patients with acute liver dysfunction. AB - OBJECTIVE: Fulminant hepatic failure (FHF) is a clinical syndrome of sudden and severe liver dysfunction accompanied by encephalopathy in a previously healthy person. In FHF, hepatocytes are severely damaged and ordinary liver regeneration is impaired. We demonstrated that the expression of osteopontin (OPN), a multifunctional cytokine, was up-regulated in mouse oval cell (a stem-cell progenitor) induction models. MATERIAL AND METHODS: Based on this finding, serum OPN levels were examined in 43 patients with FHF and in 45 patients with acute self-limited hepatitis (AH). To determine the cellular source of OPN, the expression of OPN was studied in a liver specimen derived from an FHF patient. RESULTS: The mean OPN level of patients with FHF was 2.80+/-0.48 ng/ml (log(10), +/-SD), which was significantly higher than that of the patients with AH (2.42+/ 0.39 ng/ml) (p=0.003, unpaired t-test). Patients with elevated serum OPN levels had a significantly poorer prognosis than patients whose serum OPN levels were not elevated. In the FHF patient's liver, OPN protein was expressed not only in inflammatory cells but also in regenerating hepatocytes and bile ductular structures. CONCLUSIONS: Our current study indicates that serum OPN levels increased in patients with FHF and that OPN might play an important role in liver regeneration due to activation of hepatic stem cells. PMID- 16373284 TI - Interventional endoscopic therapy in chronic pancreatitis including temporary stenting: a definitive treatment? AB - OBJECTIVE: In the past 15 years there have been tremendous advances in endoscopic management of chronic pancreatitis (CP). However, the value of endoscopic pancreatic stenting is still debatable. MATERIAL AND METHODS: In 98 patients suffering from symptomatic CP (84 M, 14 F, 49+/-12, age range 23-83 years) endotherapy including temporary stenting of the pancreatic duct was performed. After final stent removal, indicating the primary end-point of endotherapy, 96 patients were followed for 35+/-28 (8 days-111) months. All data were assessed retrospectively. RESULTS: As well as other endoscopic procedures, a total of 358 prostheses were inserted in the pancreatic duct and left in place for 3+/-1 (1 day-11) months. Total stent treatment time was 10+/-10 (6 days-49) months. At 46+/-27 (4-111) months after limited endotherapy, 57 patients had no need for secondary intervention, two-thirds were even without further pain sensations. In 22 patients, surgical treatment and in 17 patients further endoscopic therapy became necessary, which was significantly correlated with continued alcohol consumption. CONCLUSIONS: Temporary stent placement as a part of interventional endoscopic therapy in CP shows a high rate of technical and long-term clinical success, with no need for secondary treatment in a remarkable number of patients. Continued cessation of alcohol consumption supports the treatment benefit significantly. PMID- 16373285 TI - Octreotide in the treatment of refractory ascites of cirrhosis. AB - Dietary sodium restriction and diuretic treatment have been shown to be effective in the treatment of ascites in the majority of cirrhotic patients. However, approximately 5 to 10% of patients develop refractory ascites, which is defined as ascites that does not respond to intensive diuretic therapy (diuretic resistant) or ascites that cannot be controlled because the patient develops diuretic-induced complications that prevent the use of an effective diuretic dose (diuretic-intractable). Current therapeutic approaches for refractory ascites include repeated large-volume paracentesis and transjugular intrahepatic portosystemic shunting. In the present report, subcutaneous octreotide treatment improved renal function and hemodynamics and diuretic response in two patients with refractory ascites in line with a marked decrease in renin and aldosterone secretion. We consider that octreotide could be of value in the management of refractory ascites in cirrhotic patients. PMID- 16373286 TI - Gastrin vaccination against gastrointestinal and pancreatic cancer. PMID- 16373288 TI - The Nobel Prize in 2005 for the discovery of Helicobacter pylori: implications for child health. AB - The Nobel Prize in Physiology or Medicine in 2005 has been awarded to B. Marshall and R. Warren for their discovery that peptic ulcer disease is caused by an infection with Helicobacter pylori. This infection, which affects about half of the world's population and is already extremely prevalent in adolescents in developing countries, starts as an asymptomatic gastritis which, under certain conditions, is followed by gastric or duodenal ulcer disease. CONCLUSION: No proven benefit has yet been found by treating H. pylori-infected children with gastritis unless they have a peptic ulcer. Vaccination against H. pylori infection during early childhood is considered a means of preventing peptic ulcer disease and also possibly adenocarcinoma. PMID- 16373289 TI - Diagnosis and early management of inborn errors of metabolism presenting around the time of birth. AB - Inherited metabolic diseases often present around the time of birth. They are responsible for some cases of hydrops fetalis and a number of dysmorphic syndromes. Patients with inborn errors may also present at (or shortly after) birth with seizures or severe hypotonia. Most affected babies, however, appear normal at birth and subsequently deteriorate, with hypoglycaemia, acidosis, neurological or cardiac problems, or liver disease. Treatment often involves measures to reduce catabolism and to remove toxic metabolites. It should not be delayed for a definitive diagnosis. CONCLUSION: In the newborn period, inborn errors can easily be misdiagnosed as sepsis or birth asphyxia; prompt detection requires vigilance and the early measurement of biochemical markers, such as plasma ammonia. PMID- 16373290 TI - Randomized controlled trial of skin-to-skin contact from birth versus conventional incubator for physiological stabilization in 1200 g to 2199 g newborns. AB - Publication of the report of Bergman et al. should stimulate replications by others. If these investigators' findings are confirmed, and clear guidelines for the care of premature infants in the kangaroo method in the first 6 h and beyond are published, with particular attention to warm coverings for the babies' backs, this research could benefit large numbers of premature infants in the Third World. CONCLUSION: As the authors indicate, skin-to-skin contact from the birth of premature infants may also be a good alternative in First World settings. PMID- 16373291 TI - Early detection of podiatric anomalies in children with Down syndrome. AB - AIMS: To verify the importance of podiatric evaluation in patients with Down syndrome for the early diagnosis and treatment of minor orthopaedic problems. METHODS: Case-control study of 50 children affected by Down syndrome (aged 4-10 y) without major orthopaedic malformations compared to 100 healthy children. A complete podiatric examination was performed on all patients and controls. RESULTS: Children with Down syndrome showed several orthopaedic anomalies including bony deformity of the forefoot (90%), flat foot (60%), isolated calcaneal valgus (24%), knee valgus (22%) and pronated flat foot (16%). These abnormalities were responsible for postural alterations as confirmed by baropodometric examination. CONCLUSION: The data demonstrated a greater incidence of minor orthopaedic alterations and suggest the necessity of regular podiatric examinations in the follow-up of this syndrome. PMID- 16373292 TI - Assessment of total body fat using the skinfold technique in full-term and preterm infants. AB - BACKGROUND: Assessment of body composition may be of interest when the nutritional status of infants is evaluated but is often difficult since simple and valid methods are lacking. With appropriate validation, measurements based on skinfold thickness (SFT) may be useful for this purpose. AIMS: To evaluate the potential of a published method, based on measurements of SFT, to assess total body fat (TBF) of infants; and to calculate the fat content of adipose tissue (AT) in infants using previously published information regarding AT volume and total body water. SUBJECTS AND METHODS: Forty-five full-term infants and eight infants born in gestational weeks 31-33 were studied at a postnatal age of 4-131 and 44-75 d, respectively. The body water dilution method was used to obtain reference estimates of TBF (TBF-BWD). RESULTS: In full-term infants, TBF assessed using the skinfold method (TBF-SFT) minus TBF-BWD was 1.5+/-10.8% (mean+/-2 SD). Furthermore, TBF-SFT minus TBF-BWD (%, y) was correlated (p<0.0001) with the average of TBF-SFT and TBF-BWD (%, x), showing that TBF-SFT was too high in lean infants and too low in infants with more TBF. In the full-term infants, AT contained 0.68+/-0.14 g fat/ml. In the premature infants, TBF-SFT (%), TBF-BWD (%) and the AT fat content were similar to the corresponding figures in nine full term newborns. CONCLUSION: The results indicate that the SFT method produced inaccurate and biased estimates of TBF in infants. A considerable variation between infants regarding their AT fat content may be an important reason for these findings. PMID- 16373293 TI - Quality of care of the preterm infant--the parent and nurse perspective. AB - AIM: To study the subjective opinions about what is important in care at neonatal units and child health centres (CHCs) for premature newborns, and to compare these opinions with the care actually given. SUBJECTS: 21 mothers, 20 fathers and 15 nurses at the neonatal unit, and 21 mothers, 14 fathers and 18 nurses at CHCs. METHODS: A questionnaire on quality of Care from the Patient's Perspective was used. It contained three dimensions: identity-oriented approach, medical technical competence and socio-cultural atmosphere. Each dimension was evaluated in terms of subjective importance and perceived reality of given care. RESULTS: In general, subjective importance was rated higher than perceived reality both for neonatal care and care at CHCs for the dimensions identity-oriented approach and medical-technical competence. However, higher ratings were given to neonatal care compared to CHCs for medical-technical competence. High-risk diagnoses and very low gestational age in the newborn did not affect the answers. Mothers rated medical-technical competence higher than nurses for neonatal care. Mothers and nurses rated identity-oriented approach higher than fathers for CHCs. CONCLUSION: Although both neonatal care and care at CHCs were highly rated, improvements can be made to fulfil the expectations of parents and nurses. Neonatal units seem to be more efficient in taking care of the special needs of these newborns compared to CHCs. The need for an optimal identity-oriented approach, medical-technical competence and socio-cultural atmosphere could strengthen the possibilities of parents to be confident in their parental role. PMID- 16373294 TI - Increase of interleukin-6 in tracheal aspirate at birth: a predictor of subsequent bronchopulmonary dysplasia in preterm infants. AB - AIM: We tested whether interleukin-6 (IL-6) in tracheal aspirate (TA) at birth, as a marker of fetal pulmonary inflammation, can be a predictor of bronchopulmonary dysplasia (BPD) in preterm infants. METHODS: A total of 75 preterm (< or = 32 wk) infants who were intubated in the delivery room were prospectively enrolled. Multivariate logistic regression analysis was done to determine whether IL-6 in TA at birth is an independent risk factor for BPD, and a receiver-operating characteristic curve was constructed to determine the accuracy of IL-6 in TA for predicting the risk of BPD. RESULTS: IL-6 in TA at birth was an independent risk factor for BPD. Fetal pulmonary inflammation defined as IL-6 in TA at birth > or = 316 pg/ml together with patent ductus arteriosus (PDA) additively predicted the risk of BPD. The sensitivity, specificity, and positive and negative predictive values of fetal pulmonary inflammation for the identification of BPD were 73%, 71%, 58% and 83%, respectively. CONCLUSION: IL-6 in TA at birth can be used as a predictor of BPD in combination with the presence of PDA. PMID- 16373295 TI - Young children non-immunized against measles: characteristics and programmatic implications. AB - AIM: To examine the presenting characteristics, including nutritional status, of young children without measles immunization and to suggest appropriate public health measures to improve immunization status. METHODS: In this retrospective case-control analysis, we studied 4075 children aged 12-23 mo of either sex, who attended ICDDR,B's Dhaka hospital during 1994-2003. Cases included children who reported to this facility without receiving measles vaccine, and the control children were those who received the vaccine. RESULTS: 3181 of 4075 (78%) children, including 1227 (39%) girls and 1954 (61%) boys, received measles immunization. The proportion of vaccinated children increased from 74% in 1997 to 82% in 2001. Some non-specific effects of measles immunization were observed. Fifty-one per cent of the children without measles immunization were stunted, 76% were underweight, and 48% were wasted. The non-immunized children were twice as likely to be stunted, underweight, and wasted than the immunized children; they were more often dehydrated (some or severe dehydration) (28% vs 22%, p<0.001), required longer duration (>72 h) of hospitalization (15% vs 10%, p<0.001), did not receive vitamin A capsule in the previous 6 mo (56% vs 36%, p<0.001), and had more frequent abnormal lung auscultation indicative of acute lower respiratory tract infections (8% vs 5%, p<0.001). Female children, illiterate mother, lack of vitamin A supplementation, and history of measles were significantly associated with non-immunization against measles after controlling for co-variables. Results were similar when different nutritional indicators (underweight, stunting, or wasting) were added separately to logistic regression models. CONCLUSION: Intervention strategies to enhance immunization coverage in infants should target illiterate mothers and their children, particularly the females and malnourished ones, provide them with measles immunization and vitamin A capsule, and encourage their periodic follow-up visits as part of a preventive nutritional programme. PMID- 16373296 TI - The relation between inferior vena cava oxygen saturation, superior vena cava flow, fractional oxygen extraction and haemoglobin affinity in sick newborns: a pilot study. AB - AIM: To determine whether inferior vena cava oxygen saturation (UvO2) or lower body fractional oxygen extraction (FOE) could detect poor cardiac output in newborns. METHODS: UvO2 and arterial oxygen saturation (SaO2) were measured simultaneously with echocardiographic determination of superior vena cava blood flow (SVC flow) at <12, 12-24 and >24 h. Haemoglobin concentration ([Hb]), haemoglobin oxygen affinity (HOA) and lactate were measured and FOE calculated. RESULTS: 56 studies in 17 infants, gestational age (median (range)) 26 wk 4 d (23 wk 2 d-42 wk 3 d): UvO2 (mean (SD)) was 84.9% (5.0), 77.6% (9.2) and 81.7% (12.9) at <12, 12-24 and >24 h, respectively; SVC flow (mean (SD)) increased from 71.7 (33) to 85 (66) and 123 (88) ml/kg/min at <12, 12-24 and >24 h, respectively. Despite a fall in mean [Hb], mean upper-body oxygen delivery increased due to increases in both SVC flow and arteriovenous content difference. There was a negative correlation between [Hb] and FOE. Infants with high HOA had significantly lower FOE. CONCLUSION: Measurement of UvO2 is feasible in newborns. Changes to SVC flow and arteriovenous content difference lead to improvements in oxygen delivery. The interaction of HOA warrants further study. PMID- 16373297 TI - Bronchial responsiveness to methacholine and adenosine 5'-monophosphate (AMP) in young children with post-infectious bronchiolitis obliterans. AB - AIM: Bronchial hyperresponsiveness (BHR) is a characteristic feature of asthma, but it is also frequently demonstrated by children and adults with chronic obstructive lung diseases. BHR is usually measured by bronchial challenges using direct or indirect stimuli. The aim of this study was to compare these two types of bronchial challenge in young children with post-infectious bronchiolitis obliterans (BO). METHODS: Methacholine and adenosine 5'-monophosphate (AMP) bronchial challenges were performed on preschool children with post-infectious BO (n=18), those with asthma (n=23) and in controls (n=20), using a modified auscultation method. The endpoint was defined as the appearance of wheezing and/or oxygen desaturation. RESULTS: A positive response to methacholine (an endpoint concentration < or = 8 mg/ml) was observed in 88.9% (16/18) of the patients with post-infectious BO, but a positive response to AMP (an endpoint concentration < or = 200 mg/ml) was observed in only 22.2% (4/18). All patients with asthma responded positively to methacholine, and most (21/23, 91.3%) of them also responded positively to AMP. The majority of the controls were insensitive to both challenges. CONCLUSION: BHR to methacholine is a frequent, but by no means universal, finding in young children with post-infectious BO, but is usually not accompanied by BHR to AMP. PMID- 16373298 TI - Hospitalized Swedish children with hand and forearm injuries: a retrospective review. AB - AIM: To analyse incidences and find risk groups in children hospitalized with hand injuries in a national retrospective study. METHODS: All children with a primary hand or forearm injury admitted to Swedish hospitals during 1987-2001 were retrieved from the Swedish Hospital Discharge Register and analysed as to incidence and characteristics. RESULTS: Among 9855 children included, the median age was 7.0 y and two-thirds were boys. The incidence increased in both younger (0-6 y) and older (7-14 y) children. Wounds and fractures almost doubled, while muscle/tendon injuries decreased. In younger children, wounds and burns were the most frequent diagnoses. In older children, fractures and muscle/tendon injuries were common. Sharp objects caused most of the injuries. University hospitals treated almost half of the children. CONCLUSION: The number of children with hand injuries admitted to hospitals in Sweden increased annually. Analyses of causes behind the increase are important to counter this trend. PMID- 16373299 TI - Umbilical artery catheter blood sampling volume and velocity: impact on cerebral blood volume and oxygenation in very-low-birthweight infants. AB - AIM: Blood sampling from umbilical artery catheters decreases cerebral blood volume and cerebral oxygenation. The aim of this study was to assess the impact of sampling volume and velocity. METHODS: Forty-eight infants, median birthweight 965 g (480-1500 g), median gestational age 27 wk (23-34 wk), were studied during routine blood sampling from umbilical artery catheters. The sampling procedure was performed following a strict protocol for draw-up volume (1.6 ml), sampling volume (1.7 ml or 0.2 ml), re-injection volume (1.6 ml) and flushing volume (0.6 ml), time of aspiration (40 s or 80 s), re-injection (30 s) and flushing (6 s). In each infant, sampling volume and aspiration time were subject to sequential variation in a randomized fashion (1.7 ml/40 s, 1.7 ml/80 s, 0.2 ml/30 s). Using near-infrared spectroscopy, changes in concentrations of cerebral oxygenated and deoxygenated haemoglobin were measured, and changes in cerebral blood volume and cerebral oxygenation were calculated. RESULTS: During all three sampling procedures, oxygenated haemoglobin decreased significantly from baseline, whereas deoxygenated haemoglobin did not change. Correspondingly, a decrease in cerebral blood volume and cerebral oxygenation occurred. This decrease was not affected significantly by extending the sampling time from 40 s to 80 s, whereas it was blunted by reducing the amount of blood withdrawn. CONCLUSION: Blood sampling from umbilical artery catheters induces a decrease in cerebral blood volume and cerebral oxygenation. The magnitude of the decrease depends on the blood volume withdrawn but not on sampling velocity. PMID- 16373300 TI - Dexamethasone reduces reintubation rate due to postextubation stridor in a high risk paediatric population. AB - OBJECTIVE: To study the effect of dexamethasone on postextubation stridor (PS) incidence and reintubation rate due to PS in a high-risk paediatric intensive care population. PATIENTS AND METHODS: All children aged between 4 wk and 6 y, who were intubated for at least 24 h and extubated between August 1999 and May 2002, were retrospectively included (n=60). Medical records of the included patients were studied; records of patients treated with dexamethasone prior to and following extubation (n=23) were compared with control patients who had not received prophylactic medication (n=37). RESULTS: Nine patients in the control group developed significant postextubation stridor, necessitating nebulized epinephrine or glucocorticosteroids. In six of these children, reintubation as a result of postextubation stridor was indicated. None of the patients treated with dexamethasone developed severe postextubation stridor or required reintubation. CONCLUSIONS: The risk of postextubation stridor is relatively high in the group of children aged between 4 wk and 6 y with intubation exceeding 24 h. We found dexamethasone to be effective in preventing reintubation due to postextubation stridor in this paediatric high-risk group. PMID- 16373301 TI - Developmental changes in infant heart rate responses to head-up tilting. AB - AIM: Newborn infants produce significant heart rate responses to both head-up and head-down tilting: heart rate increases with head-up tilting and decreases with head-down tilting. However, previously we found that, at 2-4 mo of age, heart rate increases were no longer significant following slow head-up tilting. This study was designed to determine if 2-4-mo-old infants have significant increases in heart rate when tilted rapidly. METHODS: Fifty-four infants were tested as newborns or at 2-4 mo of age. Heart rate was measured while infants were tilted to a 30 degrees head-up angle either slowly over a period of 30 s or rapidly in 5 s. RESULTS: Newborns exhibited increases in heart rate using both tilt speeds; however, at 2-4 mo of age, heart rate did not change significantly using either speed of tilting. CONCLUSION: There are significant early developmental changes in cardiac responses to hypotensive challenge. Newborns react like adults, mounting sustained increases in heart rate in response to head-up tilting, but at 2-4 mo of age sustained heart rate responses are no longer significant. Tilt tests may provide a standardized method for assessing autonomic competence during the period of maximum vulnerability to sudden infant death syndrome. PMID- 16373302 TI - Central/diffuse abdominal perception of urological pain in children. AB - AIM: In children with recurrent abdominal pain (RAP), the location of pain to the flank may suggest the involvement of the urinary apparatus. We aimed to evaluate the location of pain in different age groups of children with idiopathic hypercalciuria (HC) and/or hyperuricosuria (HU) and RAP. METHODS: Location of pain was assessed in 180 consecutive HC/HU children with RAP, aged 3-17.6 y, presenting to a single centre. RESULTS: Considering four age groups, central/diffuse was more prevalent than lateral location of pain in youngsters, with a progressive and significant (p<0.0001, chi2 test for trend) decrease of the prevalence of central/diffuse location with rising age, and with the most relevant drop after age 8 y. Location of pain was central/diffuse in 81% of patients under and lateral in 79% of those over 8 y of age (p<0.0001). The location of pain changed from central/diffuse to lateral in 16/18 children followed at least 1 y, within 1 to 2 y by age 8. CONCLUSION: Only after 8 y of age did the lateral location of abdominal pain become the rule in children with HC/HU and RAP. This has to be kept in mind in order to avoid overlooking the urologic origin of pain. PMID- 16373303 TI - The relationship between socio-economic status, parental support and adolescent physical activity. AB - AIM: To examine: 1) the association between socio-economic status and adolescent physical activity; 2) to assess links between mothers' and fathers' social support and adolescents' physical activity. METHODS: Participants were 326 urban adolescents and their parents. Physical activity was measured using a 7-d physical activity recall. Socio-economic status and parents' social support was assessed using questionnaires. RESULTS: Boys were significantly more active than girls, and fathers' explicit modelling towards boys was higher compared to girls. In contrast, fathers' and mothers' logistic support was significantly higher towards their daughters. Social class and fathers' and mothers' social support were significantly related to adolescent self-reported physical activity. Fathers' explicit modelling was the strongest predictor of adolescent physical activity, predicting 13.5% of the total variance. Social class, fathers' logistic support and mothers' explicit modelling were also significant predictors, accounting for additional 14% of the variance in adolescents' weekly physical activity. CONCLUSION: Parental support and social class are positively related to physical activity in Estonian adolescents. PMID- 16373304 TI - Thiamine-responsive megaloblastic anaemia syndrome: long-term follow-up and mutation analysis of seven families. AB - AIM: Thiamine-responsive megaloblastic anaemia syndrome (TRMA) is the association of diabetes mellitus, anaemia and deafness, due to mutations in SLC19A2, encoding a thiamine transporter protein. This is a unique monogenic form of vitamin dependent diabetes for which there is limited long-term data. We aimed to study genotype-phenotype relationships and long-term follow-up in our cohort. METHODS: We have studied 13 patients from seven families and have follow-up data for a median of 9 y (2-30 y). RESULTS: All patients originated from Kashmir or Punjab, and presented with non-immune, insulin-deficient diabetes mellitus, sensorineural deafness and a variable anaemia in the first 5 y of life, the anaemia progressing to megaloblastic and sideroblastic changes in the bone marrow. The anaemia and diabetes mellitus responded to oral thiamine hydrochloride 25 mg/d, but during puberty thiamine supplements became ineffective, and almost all patients require insulin therapy and regular blood transfusions in adulthood. All patients are homozygous for mutations in the SLC19A2 gene. We have identified a novel missense mutation (T158R) that was excluded in 100 control alleles. CONCLUSION: Diabetes in this syndrome is due to an insulin insufficiency that initially responds to thiamine supplements; however, most patients become fully insulin dependent after puberty. A mutation screening strategy is feasible and likely to identify mutations in almost all cases. PMID- 16373305 TI - Hemoglobin A1c in obese children and adolescents who participated in a weight management program. AB - AIM: The objective of this study was to compare hemoglobin A1c values in non diabetic obese children and adolescents before enrollment and after completion of a 12-wk weight management program. METHODS: Seventeen children and adolescents, age 10.8+/-2.5 y (mean+/-1 SD), joined a multidisciplinary weight management program. Hemoglobin A1c and body mass index were measured at the start and at the completion of the program in each participant. RESULTS: Body mass index at the start of the program was 34.3+/-6.4 kg/m2. Body mass index at the end of the program was 33+/-6.6 kg/m2 (p<0.05). Hemoglobin A1c at the beginning of the program was 5.3+/-0.3%. Hemoglobin A1c at the completion of the program was 5+/ 0.2% (p<0.05). CONCLUSION: In this group of overweight children, hemoglobin A1c decreased significantly after participating in a multi-disciplinary weight loss program. These findings further point to the beneficial effect of lifestyle changes on the metabolic status of obese children. PMID- 16373306 TI - A Quisling on neonatal jaundice. AB - BACKGROUND/METHODS: Nils Andreas Quisling was born in Telemark, Norway, in 1854 and became an obstetrician. In 1893 he published a monograph on neonatal jaundice. This monograph was studied and compared with other published sources from the same time period. RESULTS: Quisling discussed four theories on the cause of neonatal jaundice. "Suppression jaundice" held that jaundice was the result of suppressed liver function. "Haematogenous jaundice" claimed that bilirubin was formed from haemoglobin outside the liver, and had nothing to do with the liver. "Polycholia" held that neonatal jaundice was due to an increased secretion of all the constituents of bile. Quisling rejected these and supported the "hepatogenous" theory-jaundice was always a liver-related phenomenon. He described 50 cases of neonatal jaundice. The incidence of neonatal jaundice at the Kristiania Maternity Foundation was 26%. In Quisling's cases, gastrointestinal disturbances were common, and he felt this to be an under reported complication of neonatal jaundice. He found that neonatal jaundice was often accompanied by fever. CONCLUSION: Gastrointestinal disturbances were causally involved in neonatal jaundice, resulting in impeded bile drainage. At the time of Quisling's study (1887), this "hepatogenous" theory of neonatal jaundice probably still had majority support in the European literature. PMID- 16373307 TI - Gallbladder cancer in a patient with Miller-Dieker syndrome. AB - We report the case of a male patient with Miller-Dieker syndrome (MDS) and gallbladder cancer. Chromosome analysis by fluorescence in situ hybridization revealed a deletion in the 17p13.3 region, an area thought to contain tumour suppressor genes, including the hypermethylated in cancer 1 gene. Considering the rarity of gallbladder cancer in children, we propose that MDS as the genetic background of this patient may have played a role in the occurrence of gallbladder cancer. CONCLUSION: Our present report indicates that the emergence of cancers should be taken into consideration during the long-term follow-up of patients with MDS. PMID- 16373308 TI - Postnatal rupture of an antenatally diagnosed choledochal cyst: first case report. AB - A 7-mo-old girl with an asymptomatic antenatally diagnosed choledochal cyst was referred to the regional paediatric surgical centre for advice on management. Following further radiological investigation and discussion with the supra regional centre, a date for elective surgery was arranged. However, in the interim, she was admitted as an emergency following rupture of the cyst. She made an uneventful recovery following urgent laparotomy. CONCLUSION: We recommend prompt paediatric surgical referral following antenatal diagnosis of a presumed choledochal cyst. Delay in surgery may permit expansion and rupture of the cyst, even after a period of asymptomatic stability. PMID- 16373309 TI - Rheumatic heart disease and coronary vasculitis in children. AB - Rheumatic fever is a systemic inflammatory disease which may also cause vasculitis in multiple organ systems. CONCLUSION: Two children with rheumatic mitral valve disease and mitral valve replacement developed myocardial ischaemia due to coronary vasculitis. One of them had neurological findings that may be attributed to cerebral vasculitis. PMID- 16373310 TI - Acute pancreatitis in paediatric systemic lupus erythematosus. AB - Acute pancreatitis (AP) rarely complicates the clinical course of systemic lupus erythematosus (SLE). AP as the initial manifestation of SLE is exceptional, but its outcome is often fatal. Corticosteroids have been suspected to play a role in the development of AP, but the therapeutic benefit seems to be far above the risk of exacerbation of pancreatic lesions. We report a 13-y-old girl presenting with arthralgia and malaise, followed by abdominal pain, generalized oedema and haemodynamic instability. Increased CRP (325 ng/ml), serum amylase (14,000 IU/l) and lipase (2500 IU/l) levels suggested AP. Acute anuric renal failure required haemodialysis. Multiorgan involvement suggested SLE, which was confirmed 3 d later by increased anti-ds-DNA levels. Three methylprednisolone pulses were administered promptly, followed by oral prednisone (1.5 mg/kg/d) and six pulses of cyclophosphamide (500 mg/1.73 m2/2 wk). Mycophenolate mofetil was introduced for long-term disease control. Amylase and lipase levels decreased over 4 wk. Renal function was normal after 3 wk and proteinuria negative after 6 wk. This case suggests that steroid pulse therapy should be promptly administered if clinical and biochemical investigations suggest SLE to be responsible for AP. Aggressive treatment may be life saving. PMID- 16373313 TI - Characteristics of job rotation in the Midwest US manufacturing sector. AB - Job rotation has been advocated as a suitable intervention to control work related musculoskeletal disorders. However, little is known regarding the prevalence of job rotation, methods used to identify jobs for rotation or the benefits or limitations of job rotation. A web-based questionnaire was developed to survey job rotation practices from Midwest US manufacturing companies. Results indicated that 42.7% of the companies contacted used job rotation, where the median time for which they had used job rotation was 5 years. Job rotation was used mainly to reduce exposure to risk factors for work-related injuries and to reduce work related injuries, whereas supervisor decisions and ergonomic analyses were used to select jobs for the rotation scheme. Major limitations to successful implementation of job rotation included rotation of individuals with medical restrictions, decreased product quality and lack of jobs to rotate to. These findings suggest that further study is needed to determine if exposure to risk factors is reduced through current efforts. PMID- 16373314 TI - Evaluating the effects of bilingual traffic signs on driver performance and safety. AB - Variable message signs (VMS) can provide immediate and relevant information to road users and bilingual VMS can provide great flexibility in countries where a significant proportion of the population speak an alternative language to the majority. The study reported here evaluates the effect of various bilingual VMS configurations on driver behaviour and safety. The aim of the study was to determine whether or not the visual distraction associated with bilingual VMS signs of different configurations (length, complexity) impacted on driving performance. A driving simulator was used to allow full control over the scenarios, road environment and sign configuration and both longitudinal and lateral driver performance was assessed. Drivers were able to read one- and two line monolingual signs and two-line bilingual signs without disruption to their driving behaviour. However, drivers significantly reduced their speed in order to read four-line monolingual and four-line bilingual signs, accompanied by an increase in headway to the vehicle in front. This implies that drivers are possibly reading the irrelevant text on the bilingual sign and various methods for reducing this effect are discussed. PMID- 16373315 TI - Interfering effects of the task demands of grip force and mental processing on isometric shoulder strength and muscle activity. AB - The purpose of this study was to examine the interfering effects of physical and mental tasks on shoulder isometric strength in different postures. Fifteen volunteers (seven women, eight men) performed a series of isometric shoulder exertions at 30 degrees , 60 degrees and 90 degrees of both shoulder flexion and abduction alone and with the addition of a 30% grip force, a mental task (Stroop test) and both additional tasks simultaneously. The shoulder tasks were completed either at maximal intensity, or while maintaining a shoulder posture without any additional effort. Surface electromyography (EMG) from seven muscles of the shoulder girdle and shoulder moment were collected for each 6 s shoulder exertion. When normalized to maximum exertion, no differences were found between genders and no differences existed between conditions when subjects maintained each posture without exerted force. In the maximal shoulder exertion trials, an increase in shoulder angle (in either plane) resulted in an increase in EMG in most muscles, while shoulder moment decreased in flexion and remained constant in abduction. Shoulder moments and muscle activation were greatest in the shoulder exertion alone condition followed by adding a 30% grip and the Stroop test, with the addition of both tasks further reducing the exerted shoulder moment and EMG. However, muscle activity did not always decrease with shoulder strength and remained elevated, indicating a complex coactivation pattern produced by an interfering role of the tasks. Overall, it was found that a mental task can have the same or greater effect as a concurrent grip and should be considered when assessing muscular loading in the workplace, as typical biomechanical modelling may underestimate internal loads. The results not only provide valuable shoulder strength data but also practical strength values, depending on additional tasks. PMID- 16373316 TI - Longitudinal changes in critical flicker fusion frequency: an indicator of human workload. AB - The aim of the study was to verify a hypothesis that critical flicker fusion frequency (CFFF) is season-related because of changes in the intensity and duration of sunlight. CFFF has a wide application in different fields, generally as an indicator of human workload. Therefore, confirmation of previous observations that CFFF is subject to a seasonal rhythm could make theoretical as well as practical sense. The analysis was based on data obtained from 23 male participants, aged 28-60 years. The study lasted 12 months. CFFF was measured with the flicker test. The results of statistical analysis indicate that CFFF changes over time. There are two forms of those changes: a linear trend and oscillations around the trend. Moreover, individual differences in the longitudinal course of CFFF (in trends as well as in oscillations) are observed. The conclusion is that CFFF is subject to longitudinal changes, whose pattern depends on individual differences. Additionally, the individual character of the CFFF changes over time allow the assumption that sunlight is probably not as significant a factor as expected, one that can modify the hypothetical pattern of season-related CFFF fluctuation. The obtained results are important for the methodology of experiments with CFFF and for the interpretation of the results of the flicker test. PMID- 16373317 TI - Effects of negatively sloped keyboard wedges on risk factors for upper extremity work-related musculoskeletal disorders and user performance. AB - Several changes to computer peripherals have been developed to reduce exposure to identified risk factors for musculoskeletal injury, notably in keyboard designs. Negative keyboard angles and their resulting effects on objective physiological measures, subjective measures and performance have been studied, although few angles have been investigated despite the benefits associated with their use. The objective of this study was to quantify the effects of negative keyboard angles on forearm muscle activity, wrist posture, key strike force, perceived discomfort and performance and to identify a negative keyboard angle or range of keyboard angles that minimizes exposure to risk factors for hand/wrist injuries. Ten experienced typists (four males and six females) participated in a laboratory study to compare keyboard angles ranging from 0 degrees to -30 degrees , at 10 degrees increments, and a keyboard with a 7 degrees slope, using a wedge designed for use with standard QWERTY keyboards. Repeatability of exposures was examined by requiring participants to complete two test sessions 1 week apart. Dependent variable data were collected during 10 min basic data entry tasks. Wrist posture data favoured negative keyboard angles of 0 degrees (horizontal) or greater, compared to a positive keyboard angle of 7 degrees , especially for the flexion/extension direction. In general, the percentage of wrist movements within a neutral zone and the percentages of wrist movements within +/-5 degrees and +/ 10 degrees increased as keyboard angle became more negative. Electromyography results were mixed, with some variables supporting negative keyboard angles whilst other results favoured the standard keyboard configuration. Net typing speed supported the -10 degrees keyboard angle, whilst other negative typing angles were comparable, if not better than, with the standard keyboard. Therefore, angles ranging from 0 degrees to -30 degrees in general provide significant reductions in exposure to deviated wrist postures and muscle activity and comparable performance. PMID- 16373318 TI - Bacterial species selective toxicity of two isomeric alpha/beta-peptides: role of membrane lipids. AB - We have studied how membrane interactions of two synthetic cationic antimicrobial peptides with alternating alpha- and beta-amino acid residues ("alpha/beta peptides") impact toxicity to different prokaryotes. Electron microscopic examination of thin sections of Escherichia coli and of Bacillus subtilis exposed to these two alpha/beta-peptides reveals different structural changes in the membranes of these bacteria. These two peptides also have very different effects on the morphology of liposomes composed of phosphatidylethanolamine and phosphatidylglycerol in a 2:1 molar ratio. Freeze fracture electron microscopy indicates that with this lipid mixture, alpha/beta-peptide I induces the formation of a sponge phase. 31P NMR and X-ray diffraction are consistent with this conclusion. In contrast, with alpha/beta-peptide II and this same lipid mixture, a lamellar phase is maintained, but with a drastically reduced d spacing. alpha/beta-Peptide II is more lytic to liposomes composed of these lipids than is I. These findings are consistent with the greater toxicity of alpha/beta-peptide II, relative to alpha/beta-peptide I, to E. coli, a bacterium having a high content of phosphatidylethanolamine. In contrast, both alpha/beta peptides display similar toxicity toward B. subtilis, in accord with the greater anionic lipid composition in its membrane. This work shows that variations in the selectivity of these peptidic antimicrobial peptides toward different strains of bacteria can be partly determined by the lipid composition of the bacterial cell membrane. PMID- 16373319 TI - Fluorescence and molecular dynamics studies of the acetylcholine receptor gammaM4 transmembrane peptide in reconstituted systems. AB - A combination of fluorescence spectroscopy and molecular dynamics (MD) is applied to assess the conformational dynamics of a peptide making up the outermost ring of the nicotinic acetylcholine receptor (AChR) transmembrane region and the effect of membrane thickness and cholesterol on the hydrophobic matching of this peptide. The fluorescence studies exploit the intrinsic fluorescence of the only tryptophan residue in a synthetic peptide corresponding to the fourth transmembrane domain of the AChR gamma subunit (gammaM4-Trp(6)) reconstituted in lipid bilayers of varying thickness, and combine this information with quenching studies using depth-sensitive phosphatidylcholine spin-labeled probes and acrylamide, polarization of fluorescence, and generalized polarization of Laurdan. A direct correlation was found between bilayer width and the depth of insertion of Trp(6). We further extend our recent MD study of the conformational dynamics of the AChR channel to focus on the crosstalk between M4 and the lipid belt region. The isolated gammaM4 peptide is shown to possess considerable orientational flexibility while maintaining a linear alpha-helical structure, and to vary its tilt depending on bilayer width and cholesterol (Chol) content. MD studies also show that gammaM4 also establishes contacts with the other TM peptides on its inner face, stabilizing a shorter TM length that is still highly sensitive to the lipid environment. In the native membrane the topology of the M4 ring is likely to exhibit a similar behavior, dynamically modifying its tilt to match the hydrophobic thickness of the bilayer. PMID- 16373320 TI - Structural and dynamic studies of the gamma-M4 trans-membrane domain of the nicotinic acetylcholine receptor. AB - A structural characterization of a synthetic peptide corresponding to the fourth transmembrane domain (M4-TMD) of the gamma-subunit of the nicotinic acetylcholine receptor from Torpedo californica has been undertaken. Solid-state NMR and CD spectroscopy studies indicate that upon reconstitution into lipid vesicles or magnetically aligned lipid bilayers, the synthetic M4-TMD adopts a linear alpha helical conformation with the helix aligned within 15 degrees of the membrane normal. Furthermore, analysis of the motional averaging of anisotropic interactions present in the solid-state NMR spectra of the reconstituted peptide, indicate that the dynamics of the peptide within the bilayer are highly sensitive to the phase adopted by the lipid bilayer, providing an insight into how the interaction of lipids with this domain may play a important role in the modulation of this receptor by its lipid environment. PMID- 16373321 TI - Functional characterization of P2X3 receptors fused with fluorescent proteins. AB - P2X receptor function in the CNS is poorly understood, and currently available data are partly inconsistent. In the presented study, we investigated P2X3 receptors stably expressed in HEK293 cells. Non-stationary noise analysis of whole cell currents and rapid ATP application through flash photolysis allowed for assessing the single channel conductance (6.6 pS) and the fast activation kinetics of the receptor (20 ms). The characteristics of channel desensitization and pharmacological properties matched previous findings. The properties of wild type receptors were compared with P2X3 constructs carrying a fluorescent tag (ECFP or DsRed2) at the C-terminus. These fluorescently labeled subunits formed functional receptors, with neither the affinity of the ligand binding site nor channel properties (ion selectivity, gating kinetics, single channel conductance) differing from wild type. We conclude that both fusion proteins tested here are suitable for generating transgenic mice, which can be expected to promote understanding of the physiological role of P2X3 receptors in CNS signaling. PMID- 16373322 TI - GPCR-induced dissociation of G-protein subunits in early stage signal transduction. AB - G-protein coupled receptors (GPCRs) form a ternary complex of agonist, receptor and G-proteins during primary signal transduction at the cell membrane. Downstream signalling is thought to be preceded by the process of dissociation of Galpha and Gbetagamma subunits, thus exposing new surfaces to interact with downstream effectors. We demonstrate here for the first time, the dissociation of heterotrimeric G-protein subunits (i.e., Galpha and Gbetagamma) following agonist induced GPCR (alpha(2A)-adrenergic receptor; alpha(2A)-AR) activation in a cell free assay system. alpha(2A)-AR membranes were reconstituted with the G-proteins (+/-hexahistidine-tagged) Galpha(i1) and Gbeta1gamma2 and functional signalling was determined following activation of the reconstituted receptor:G-protein complex with the potent agonist UK-14304, and [35S]GTPgammaS. In the presence of Ni(2+)-coated agarose beads, the activated his-tagged Galpha(i1)his [35S]GTPgammaS complex was captured on the Ni(2+)-presenting surface. When his tagged Gbeta1gamma2 (Gbeta1gamma2his) was used with Galpha(i1), the [35S]GTPgammaS-bound Galpha(i1) was not present on the Ni(2+)-coated beads, but rather, it was separated from the beta1gamma2(his)-beads, demonstrating receptor induced dissociation of Galpha and Gbetagamma subunits. Treatment of the reconstituted alpha(2A)-AR membranes containing Gbeta1gamma2his:Galpha(i1) with imidazole confirmed the specificity for the Ni2+:G-protein surface dissociation of Galpha(i1) from Gbeta1gamma2his. These data demonstrate for the first time, the complete dissociation of the G-protein subunits and extend observations on the role of G-proteins in the assembly and disassembly of the ternary complex in the primary events of GPCR signalling. PMID- 16373323 TI - Activation of a mechanosensitive BK channel by membrane stress created with amphipaths. AB - Some BK channels are activated in response to membrane stretch. However, it remains largely unknown which membrane component transmits forces to the channel and which part of the channel senses the force. Recently, we have shown that a BK channel cloned from chick heart (named SAKCa channel) is a stretch activated channel, while deletion of a 59 amino acids splice insert (STREX) located in the cytoplasmic side, abolishes its stretch-sensitivity. This finding raised a question whether stress in the bilayer is crucial for the mechanical activation of the channel. To address this question we examined the effects of membrane perturbing amphipaths on the stretch activation of the SAKCa channel and its STREX-deletion mutant. We found that both anionic amphipath trinitrophenol (TNP) and cationic amphipath chlorpromazine (CPZ) could dose-dependently activate the channel by leftward shifting the voltage activation curve when applied alone. In contrast, TNP and CPZ compensated each other's effect when applied sequentially. These results can be understood in the framework of the bilayer couple hypothesis, suggesting that stress in the plasma membrane can activate the SAKCa channel. Interestingly, the STREX-deletion mutant channel has much less sensitivity to the amphipaths, suggesting that STREX acts as an intermediate structure that can indirectly convey stress in the membrane to the gate of the SAKCa channel via an unidentified membrane associated protein(s) that can detect or transmit stress in the membrane. PMID- 16373324 TI - Thyroid hormones differentially regulate the distribution of rabbit skeletal muscle Ca(2+)-ATPase (SERCA) isoforms in light and heavy sarcoplasmic reticulum. AB - The sarcoplasmic reticulum (SR) is composed of two fractions, the heavy fraction that contains proteins involved in Ca2+ release, and the light fraction enriched in Ca(2+)-ATPase (SERCA), an enzyme responsible for Ca2+ transport from the cytosol to the lumen of SR. It is known that in red muscle thyroid hormones regulate the expression of SERCA 1 and SERCA 2 isoforms. Here we show the effects of thyroid hormone on SERCA expression and distribution in light and heavy SR fractions from rabbit white and red muscles. In hyperthyroid red muscle there is an increase of SERCA 1 and a decrease of SERCA 2 expression. This is far more pronounced in the heavy than in the light SR fraction. As a result, the rates of Ca(2+)- ATPase activity and Ca(2+)-uptake by the heavy vesicles are increased. In hypothyroidism we observed a decrease in SERCA 1 and no changes in the amount of SERCA 2 expressed. This promoted a decrease of both Ca(2+)-uptake and Ca(2+) ATPase activity. While the major differences in hyperthyroidism were found in the heavy SR fraction, the effects of hypothyroidism were restricted to light SR fraction. In white muscle we did not observe any significant changes in either hypo- or hyperthyroidism in both SR fractions. Thus, the regulation of SERCA isoforms by thyroid hormones is not only muscle specific but also varies depending on the subcellular compartment analyzed. These changes might correspond to the molecular basis of the altered contraction and relaxation rates detected in thyroid dysfunction. PMID- 16373325 TI - Membrane organization of the human serotonin(1A) receptor monitored by detergent insolubility using GFP fluorescence. AB - Insolubility in non-ionic detergents such as Triton X-100 at low temperature is a widely used biochemical criterion for characterization of membrane domains. In view of the emerging role of membrane organization in the function of G-protein coupled receptors, we have examined detergent insolubility of the 5-HT(1A) receptor in CHO cells using a novel GFP fluorescence approach developed by us. Using this approach, we have explored the membrane organization of the serotonin(1A) receptor tagged to enhanced yellow fluorescent protein (5-HT(1A)R EYFP) stably expressed in CHO-K1 cells under conditions of varying detergent concentration, reduced membrane cholesterol and agonist stimulation. Our results show that a small yet significant fraction of the 5-HT(1A) receptor exhibits detergent insolubility, which increases upon depletion of membrane cholesterol. Stimulation of 5-HT(1A)R-EYFP by its endogenous ligand, serotonin, did not cause a significant change in the detergent insolubility of the receptor. Taken together, our results on detergent insolubility of 5-HT(1A)R-EYFP provide new insights into the membrane organization of the 5-HT(1A) receptor and could be relevant in the analysis of membrane organization of other G-protein coupled receptors. PMID- 16373326 TI - Isolation and function of the amino acid transporter PAT1 (slc36a1) from rabbit and discrimination between transport via PAT1 and system IMINO in renal brush border membrane vesicles. AB - Reabsorption of amino acids is an important function of the renal proximal tubule. pH-dependent amino acid transport has been measured previously using rabbit renal brush-border membrane vesicles (BBMV). The purpose of this investigation was to determine whether this pH-dependent uptake represents H(+)/amino acid cotransport via a PAT1-like transport system. The rabbit PAT1 cDNA was isolated (2296bp including both 5' and 3' untranslated regions and poly(A) tail) and the open reading frame codes for a protein of 475 amino acids (92% identity to human PAT1). Rabbit PAT1 mRNA was found in all tissues investigated including kidney. When expressed heterologously in a mammalian cell line, rabbit PAT1 mediates pH-dependent, Na(+)-independent uptake of proline, glycine, l-alanine and alpha-(methylamino)isobutyric acid. Proline uptake was maximal at pH 5.0 (K(m) 2.2+/-0.7 mM). A transport system with identical characteristics (ion dependency, substrate specificity) was detected in rabbit renal BBMV where an overshoot was observed in the absence of Na+ but in the presence of an inwardly directed H+ gradient. In the presence of Na+ and under conditions in which PAT1 transport function was suppressed, a second proline uptake system was detected that exhibited functional characteristics similar to those of the IMINO system. The functional characteristics of rabbit PAT1 in either mammalian cells or renal BBMV suggest that PAT1 is the low-affinity transporter of proline, glycine and hydroxyproline believed to be defective in patients with iminoglycinuria. PMID- 16373327 TI - Atypical uses of atypical antipsychotics. AB - Atypical antipsychotic drugs are primarily indicated for the treatment of psychotic disorders such as schizophrenia and schizoaffective disorder. Recently, they have also been used for mood stabilization. This article reviews other, potentially therapeutically useful indications for these medications. In most cases, the evidence supporting these new uses is limited but provocative, and involves only case reports. It has not yet been determined whether the usefulness of atypical antipsychotics for nonpsychotic disorders outweighs their potential to cause serious side effects. PMID- 16373328 TI - Atypical antipsychotics in elderly patients with dementia or schizophrenia: review of recent literature. AB - Atypical antipsychotics have become a common pharmacologic option for the treatment of various psychiatric and behavioral symptoms in older adults, although these medications have been officially approved by the U.S. Food and Drug Administration for use only in schizophrenia and bipolar disorder. Despite the widespread use of these agents, there is a relative shortage of rigorously conducted trials. This review focuses on recently published randomized, blinded, controlled trials involving the use of atypical antipsychotics in elderly patients with dementia (n = 9) or schizophrenia (n = 3), with some discussion of published large, open-label studies and a few unpublished controlled trials. In general, the studies of patients with dementia reported modest efficacy of atypical antipsychotics when compared to placebo and conventional antipsychotics. In addition, an advantage in terms of motor side effects was consistently noted with atypical antipsychotics when compared to conventional antipsychotics. The studies have also shown, however, a greater risk of mortality and adverse cerebrovascular events with several of these agents than with placebo in individuals with dementia. There are insufficient data comparing atypical antipsychotics to one another. In the trials involving elderly persons with schizophrenia, atypical antipsychotics were associated with significant improvements in psychopathology; differences in efficacy among atypical antipsychotics were unclear. A careful consideration of the risk-benefit ratio of atypical antipsychotics, as well as that of available alternative treatments, is needed for each individual elderly patient. Clinical judgment, caution, and consent should be the watchwords in this area of psychopharmacology. PMID- 16373329 TI - Pharmacologic implications of neurobiological models of schizophrenia. AB - The dopamine model of schizophrenia has been supplanted by several additional models in order to account for genetic findings, risk factors, course of illness, and the diversity of symptom domains. The increasing number and complexity of potential models for this heterogeneous disorder offer new targets for pharmacologic treatment that differ in their appropriate time points for intervention and in their potential effects on the course of illness. This article reviews relevant neurodevelopmental, biochemical, and neurodegenerative models with respect to potential pharmacologic opportunities. PMID- 16373330 TI - Immunoglobulin G specifically binding plant N-glycans with high affinity could be generated in rabbits but not in mice. AB - Xylosylated and core alpha1,3-fucosylated N-glycans from plants are immunogenic, and they play a still obscure role in allergy and in the field of plant-made protein pharmaceuticals. We immunized mice to generate monoclonal antibodies (mAbs) binding plant N-glycans specifically via the epitope containing either the xylose or the core alpha1,3-fucose residue. Splenocytes expressing N-glycan specific antibodies derived from C57BL/6 mice previously immunized with plant glycoproteins were preselected by cell sorting to generate hybridoma lines producing specific antibodies. However, we obtained only mAbs unable to distinguish fucosylated from xylosylated N-glycans and reactive even with the pentasaccharide core Man3GlcNAc2. In contrast, immunization of rabbits yielded polyclonal sera selectively reactive with either fucosylated or xylosylated N glycans. Purification of these sera using glyco-modified neoglycoproteins coupled to a chromatography matrix provided polyclonal sera suitable for affinity determination. Surface plasmon resonance measurements using sensor chips with immobilized glyco-modified transferrins revealed dissociation constants of around 10(-9) M. This unexpectedly high affinity of IgG antibodies toward carbohydrate epitopes has repercussions on our conception of the binding strength and significance of antiglycan IgE antibodies in allergy. PMID- 16373331 TI - Shielding of the A1 domain by the D'D3 domains of von Willebrand factor modulates its interaction with platelet glycoprotein Ib-IX-V. AB - Soluble von Willebrand factor (VWF) has a low affinity for platelet glycoprotein (GP) Ibalpha and needs immobilization and/or high shear stress to enable binding of its A1 domain to the receptor. The previously described anti-VWF monoclonal antibody 1C1E7 enhances VWF/GPIbalpha binding and recognizes an epitope in the amino acids 764-1035 region in the N-terminal D'D3 domains. In this study we demonstrated that the D'D3 region negatively modulates the VWF/GPIb-IX-V interaction; (i) deletion of the D'D3 region in VWF augmented binding to GPIbalpha, suggesting an inhibitory role for this region, (ii) the isolated D'D3 region inhibited the GPIbalpha interaction of a VWF deletion mutant lacking this region, indicating that intramolecular interactions limit the accessibility of the A1 domain, (iii) using a panel of anti-VWF monoclonal antibodies, we next showed that the D'D3 region is in close proximity with the A1 domain in soluble VWF but not when VWF was immobilized; (iv) destroying the epitope of 1C1E7 resulted in a mutant VWF with an increased affinity for GPIbalpha. Our results support a model of domain translocation in VWF that allows interaction with GPIbalpha. The suggested shielding interaction of the A1 domain by the D'D3 region then becomes disrupted by VWF immobilization. PMID- 16373332 TI - Inhibition or activation of Apert syndrome FGFR2 (S252W) signaling by specific glycosaminoglycans. AB - Most Apert syndrome patients harbor a single amino acid mutation (S252W) in fibroblast growth factor (FGF) receptor 2 (FGFR2), which leads to abnormal FGF/FGFR2 signaling. Here we show that specific combinations of FGFs and glycosaminoglycans activate both alternative splice forms of the mutant but not of the wild-type FGF receptors. More importantly, 2-O- and N-sulfated heparan sulfate, prepared by a combined chemical and enzymatic synthesis, antagonized the over-activated FGFR2b (S252W) to basal levels at nanomolar concentrations. These studies demonstrated that specific glycosaminoglycans could be useful in treating ligand-dependent FGFR signaling-related diseases, such as Apert syndrome and cancer. PMID- 16373333 TI - HOXA1 is required for E-cadherin-dependent anchorage-independent survival of human mammary carcinoma cells. AB - Forced expression of HOXA1 is sufficient to stimulate oncogenic transformation of immortalized human mammary epithelial cells and subsequent tumor formation. We report here that the expression and transcriptional activity of HOXA1 are increased in mammary carcinoma cells at full confluence. This confluence dependent expression of HOXA1 was abrogated by incubation of cells with EGTA to produce loss of intercellular contact and rescued by extracellular addition of Ca2+. Increased HOXA1 expression at full confluence was prevented by an E cadherin function-blocking antibody and attachment of non-confluent cells to a substrate by homophilic ligation of E-cadherin increased HOXA1 expression. E cadherin-directed signaling increased HOXA1 expression through Rac1. Increased HOXA1 expression consequent to E-cadherin-activated signaling decreased apoptotic cell death and was required for E-cadherin-dependent anchorage-independent proliferation of human mammary carcinoma cells. HOXA1 is therefore a downstream effector of E-cadherin-directed signaling required for anchorage-independent proliferation of mammary carcinoma cells. PMID- 16373334 TI - Direct regulation of myelin protein zero expression by the Egr2 transactivator. AB - During myelination of the peripheral nervous system, the myelin protein zero (Mpz) gene is induced to produce the most abundant protein component (P(0)) of mature myelin. Although the basal embryonic expression of Mpz in Schwann cells has been attributed to regulation by Sox10, the molecular mechanism for the profound up-regulation of this gene during myelination has not been established. In this study, we have identified a highly conserved element within the first intron of the Mpz gene, which contains binding sites for the early growth response 2 (Egr2/Krox20) transcription factor, a critical regulator of peripheral nerve myelination. Egr2 can transactivate the intron element, and the induction is blocked by two known repressors of Egr2 activity. Using chromatin immunoprecipitation assays, we find that Egr2 binds in vivo to the intron element, but not to the Mpz promoter. Known inducers of Mpz expression such as forskolin and insulin-like growth factor-1 also activate the element in an Egr2 dependent manner. In addition, we found that Egr2 can act synergistically with Sox10 to activate this intron element, suggesting a model in which cooperative interactions between Egr2 and Sox10 mediate a large increase in Mpz expression to the high levels found in myelinating Schwann cells. PMID- 16373335 TI - RUNX3 cooperates with FoxO3a to induce apoptosis in gastric cancer cells. AB - The transcription factor RUNX3, which mediates apoptosis and cell growth inhibition in gastric epithelial cells, is a candidate tumor suppressor that is frequently lost in gastric cancer cells. Here, we found that restoration of RUNX3 expression in the cell line not expressing RUNX3 induced apoptosis and that it physically interacted with the Forkhead transcription factor FoxO3a/FKHRL1, known to be an important regulator of apoptosis and the cell cycle. Active unphosphorylated FoxO3a/FKHRL1 was expressed in the gastric cancer cell lines. RUNX3-induced apoptosis depended on the expression of Bim, a proapoptotic BH3 only protein, and both RUNX3 and FoxO3a/FKHRL1 were required for induction of Bim expression. Furthermore, we showed that interaction of RUNX3 and FoxO3a/FKHRL1 was also indispensable for Bim expression and apoptosis in mouse embryonic fibroblasts. In the Bim promoter, RUNX3 bound to two conserved RUNX-binding elements (RBE1 and RBE2), with RBE1 being immediately downstream of a FoxO binding element. The physical interaction of RUNX3 and FoxO3a/FKHRL1 on the Bim promoter activated transcription of Bim. These findings show that RUNX3 cooperates with FoxO3a/FKHRL1 to participate in the induction of apoptosis by activating Bim and may play an important role in tumor suppression in gastric cancer. PMID- 16373336 TI - Endogenous and exogenous Ca2+ buffers differentially modulate Ca2+-dependent inactivation of Ca(v)2.1 Ca2+ channels. AB - Voltage-gated Ca2+ channels undergo a negative feedback regulation by Ca2+ ions, Ca2+-dependent inactivation, which is important for restricting Ca2+ signals in nerve and muscle. Although the molecular details underlying Ca2+-dependent inactivation have been characterized, little is known about how this process might be modulated in excitable cells. Based on previous findings that Ca2+ dependent inactivation of Ca(v)2.1 (P/Q-type) Ca2+ channels is suppressed by strong cytoplasmic Ca2+ buffering, we investigated how factors that regulate cellular Ca2+ levels affect inactivation of Ca(v)2.1 Ca2+ currents in transfected 293T cells. We found that inactivation of Ca(v)2.1 Ca2+ currents increased exponentially with current amplitude with low intracellular concentrations of the slow buffer EGTA (0.5 mm), but not with high concentrations of the fast Ca2+ buffer BAPTA (10 mm). However, when the concentration of BAPTA was reduced to 0.5 mm, inactivation of Ca2+ currents was significantly greater than with an equivalent concentration of EGTA, indicating the importance of buffer kinetics in modulating Ca2+-dependent inactivation of Ca(v)2.1. Cotransfection of Ca(v)2.1 with the EF-hand Ca2+-binding proteins, parvalbumin and calbindin, significantly altered the relationship between Ca2+ current amplitude and inactivation in ways that were unexpected from behavior as passive Ca2+ buffers. We conclude that Ca2+ dependent inactivation of Ca(v)2.1 depends on a subplasmalemmal Ca2+ microdomain that is affected by the amplitude of the Ca2+ current and differentially modulated by distinct Ca2+ buffers. PMID- 16373337 TI - Selective permeability of different connexin channels to the second messenger cyclic AMP. AB - Gap junctions are intercellular conduits that are formed in vertebrates by connexin proteins and allow diffusion exchange of intracellular ions and small molecules. At least 20 different connexin genes in the human and mouse genome are cell-type specifically expressed with overlapping expression patterns. A possible explanation for this diversity could be different permeability of biologically important molecules, such as second messenger molecules. We have recently demonstrated that cyclic nucleotide-gated channels can be used to quantify gap junction-mediated diffusion of cyclic AMP. Using this method we have compared the relative permeability of gap junction channels composed of connexin 26, 32, 36, 43, 45, or 47 proteins toward the second messenger cAMP. Here we show that cAMP permeates through the investigated connexin channels with up to 30-fold different efficacy. Our results suggest that intercellular cAMP signaling in different cell types can be affected by the connexin expression pattern. PMID- 16373338 TI - A novel receptor activator of NF-kappaB (RANK) cytoplasmic motif plays an essential role in osteoclastogenesis by committing macrophages to the osteoclast lineage. AB - Receptor activator of NF-kappaB (RANK) ligand (RANKL) and its receptor RANK play an essential role in osteoclastogenesis and osteoclast function by activating several signaling pathways. However, several lines of evidence suggest that RANK also transmits an unidentified signaling pathway(s) essential for osteoclastogenesis. To identify the novel pathway(s), we carried out a detailed structure/function study of the RANK cytoplasmic domain. A series of studies using numerous deletion/point mutants elucidated a specific 4-amino acid motif (535IVVY538) essential for osteoclastogenesis. This novel motif plays a crucial role in committing macrophages to the osteoclast lineage but is not implicated in osteoclast function or survival. Moreover, this motif does not activate the known RANK signaling pathways, indicating that it initiates a novel pathway(s). The identification of the novel motif not only provides critical insight into RANK signaling in osteoclastogenesis, but more importantly, the RANK motif and its downstream signaling pathways may represent specific therapeutic targets for various bone diseases, including postmenopausal osteoporosis. PMID- 16373339 TI - OAZ regulates bone morphogenetic protein signaling through Smad6 activation. AB - The intensity and duration of activation of a signal transduction system are important determinants of the specificity of the cellular response to the stimulus. It is unclear how different cells can generate a signal of varying intensity and duration in response to the same cytokine. We investigated the role of the transcriptional activator and Smad1/4 cofactor OAZ in regulating bone morphogenetic protein (BMP) signaling. We demonstrate that upon BMP4 stimulation, an OAZ-Smad1/4 complex binds to and activates the gene encoding Smad6, a specific inhibitor of the BMP pathway. Removal of endogenous OAZ from pluripotent embryonal carcinoma cells prevents the induction of Smad6 by BMP4 and extends the period of detection of phosphorylated Smad1 after BMP stimulation. Conversely, in cells that do not normally express OAZ, such as myoblasts and smooth muscle cells, forced OAZ expression leads to faster and higher Smad6 induction in response to BMP4, decrease of Smad1 phosphorylation, and attenuation of BMP mediated responses. Our results demonstrate that OAZ can alter the intensity and duration of the BMP stimulus through Smad6 and indicate that the tissue-specific expression of OAZ is a critical determinant of the cellular response to the BMP signal. PMID- 16373340 TI - Forskolin-induced cell shrinkage and apical translocation of functional enhanced green fluorescent protein-human alphaENaC in H441 lung epithelial cell monolayers. AB - Elevation of intracellular cAMP increases fluid re-absorption in the lung by raising amiloride-sensitive Na+ transport through the apically localized epithelial, amiloride-sensitive Na+ channel (ENaC). However, the signaling pathways mediating this response are still not fully understood. We show that inhibition of protein-tyrosine kinase (PTK) with Genistein and protein kinase A (PKA) with KT5720, decreased forskolin-stimulated amiloride-sensitive short circuit current (I(sc)) across H441 adult human lung epithelial cell monolayers. KT5720 also decreased basal I(sc). Stable expression of green fluorescent protein (GFP)-labeled human alphaENaC in H441 cells was used to investigate dynamic changes in the cellular localization of this protein in response to forskolin. Reverse transcription-PCR and immunoblotting analysis revealed two clones expressing a truncated (alphaC3-5) and full-length (alphaC3-3) EGFP-halphaENaC protein. Only the alphaC3-3 clone displayed dome formation and exhibited a 50% increase in basal and forskolin-stimulated amiloride-sensitive I(sc) indicating that the full-length protein was required for functional activity. Apical surface biotinylation and real-time confocal microscopy demonstrated that EGFP-halphaENaC (alphaC3-3) translocated to the apical membrane in response to forskolin in a Brefeldin A-sensitive manner. This effect was completely inhibited by Genistein but only partially inhibited by KT5720. Forskolin also induced a reduction in the height of cells within alphaC3-3 monolayers, indicative of cell shrinkage. This effect was inhibited by KT5720 but not by Genistein or Brefeldin A. These data show that forskolin activates PKA-sensitive cell shrinkage in adult human H441 lung epithelial cell monolayers, which induces a PTK-sensitive translocation of EGFP-halphaENaC subunits to the apical membrane and increases amiloride-sensitive Na+ transport. PMID- 16373341 TI - CREB-AP1 protein complexes regulate transcription of the collagen XXIV gene (Col24a1) in osteoblasts. AB - Collagen XXIV is a newly discovered and poorly characterized member of the fibril forming family of collagen molecules, which displays unique structural features of invertebrate fibrillar collagens and is expressed predominantly in bone tissue. Here we report the characterization of the proximal promoter of the mouse gene (Col24a1) and its regulation in osteoblastic cells. Using well characterized murine models of osteoblast differentiation, we found that the Col24a1 gene is activated sometime before onset of the late differentiation marker osteocalcin. Additional analyses revealed that Col24a1 produces equal amounts of two alternatively spliced products with different 5'-untranslated sequences that originate from distinct transcriptional start sites. Cell transfection experiments in combination with DNA binding assays demonstrated that Col24a1 promoter activity in ROS17/2.8 osteosarcoma cells is under the control of an upstream cis-acting element, which is shared by both transcripts and is recognized by specific combinations of c-Jun, CREB1, ATF1, and ATF2 dimers. Consistent with these results, overexpression of c-Jun, ATF1, ATF2, or CREB1 in transiently transfected osteoblastic cells stimulated transcription from reporter gene constructs driven by the Col24a1 promoter to different degrees. Moreover, chromatin immunoprecipitation experiments showed that these nuclear factors bind the same upstream sequence of the endogenous Col24a1 gene. Collectively these data provide new information about transcriptional control of collagen fibrillogenesis, in addition to implicating for the first time CREB-AP1 protein complexes in the regulation of collagen gene expression in osteoblasts. PMID- 16373342 TI - Tumor necrosis factor promotes Runx2 degradation through up-regulation of Smurf1 and Smurf2 in osteoblasts. AB - Tumor necrosis factor (TNF) plays an important role in the pathogenesis of inflammatory bone loss through stimulation of osteoclastic bone resorption and inhibition of osteoblastic bone formation. Compared with the well established role of TNF in osteoclastogenesis, mechanisms by which TNF inhibits osteoblast function have not been fully determined. Runx2 is an osteoblast-specific transcription factor whose steady-state protein levels are regulated by proteasomal degradation, mediated by the E3 ubiquitin ligases, Smurf1 and Smurf2. We hypothesized that TNF inhibits osteoblast function through Smurf-mediated Runx2 degradation. We treated C2C12 and 2T3 osteoblast precursor cell lines and primary osteoblasts with TNF and found that TNF, but not interleukin-1, significantly increased Smurf1 and Smurf2 expression. TNF increased the degradation of endogenous or transfected Runx2 protein, which was blocked by treating cells with a proteasomal inhibitor or by infecting cells with small interfering (si)RNA against Smurf1 or Smurf2. TNF inhibited the expression of bone morphogenetic protein and transforming growth factor-beta signaling reporter constructs, and the inhibition of each was blocked by Smurf1 siRNA and Smurf2 siRNA, respectively. Overexpression of Smurf1 and/or Smurf2 siRNAs prevented the inhibitory effect of TNF on Runx2 reporter. Consistent with these in vitro findings, bones from TNF transgenic mice or TNF-injected wild type mice had increased Smurf1 and decreased Runx2 protein levels. We propose that one of the mechanisms by which TNF inhibits bone formation in inflammatory bone disorders is by promoting Runx2 proteasomal degradation through up-regulation of Smurf1 and Smurf2 expression. PMID- 16373343 TI - Uncoupled redox systems in the lumen of the endoplasmic reticulum. Pyridine nucleotides stay reduced in an oxidative environment. AB - The redox state of the intraluminal pyridine nucleotide pool was investigated in rat liver microsomal vesicles. The vesicles showed cortisone reductase activity in the absence of added reductants, which was dependent on the integrity of the membrane. The intraluminal pyridine nucleotide pool could be oxidized by the addition of cortisone or metyrapone but not of glutathione. On the other hand, intraluminal pyridine nucleotides were slightly reduced by cortisol or glucose 6 phosphate, although glutathione was completely ineffective. Redox state of microsomal protein thiols/disulfides was not altered either by manipulations affecting the redox state of pyridine nucleotides or by the addition of NAD(P)+ or NAD(P)H. The uncoupling of the thiol/disulfide and NAD(P)+/NAD(P)H redox couples was not because of their subcompartmentation, because enzymes responsible for the intraluminal oxidoreduction of pyridine nucleotides were distributed equally in smooth and rough microsomal subfractions. Instead, the phenomenon can be explained by the negligible representation of glutathione reductase in the endoplasmic reticulum lumen. The results demonstrated the separate existence of two redox systems in the endoplasmic reticulum lumen, which explains the contemporary functioning of oxidative folding and of powerful reductive reactions. PMID- 16373344 TI - Dynamic ligation properties of the Escherichia coli heme chaperone CcmE to non covalently bound heme. AB - The cytochrome c maturation protein CcmE is an essential membrane-anchored heme chaperone involved in the post-translational covalent attachment of heme to c type cytochromes in Gram-negative bacteria such as Escherichia coli. Previous in vitro studies have shown that CcmE can bind heme both covalently (via a histidine residue) and non-covalently. In this work we present results on the latter form of heme binding to a soluble form of CcmE. Examination of a number of site directed mutants of E. coli CcmE by resonance Raman spectroscopy has identified ligands of the heme iron and provided insight into the initial steps of heme binding by CcmE before it binds the heme covalently. The heme binding histidine (His-130) appears to ligate the heme iron in the ferric oxidation state, but two other residues ligate the iron in the ferrous form, thereby freeing His-130 to undergo covalent attachment to a heme vinyl group. It appears that the heme ligation in the non-covalent form is different from that in the holo-form, suggesting that a change in ligation could act as a trigger for the formation of the covalent bond and showing the dynamic and oxidation state-sensitive ligation properties of CcmE. PMID- 16373345 TI - Structure-function studies of two synthetic anti-vascular endothelial growth factor Fabs and comparison with the Avastin Fab. AB - In the quest to discover new research tools and to develop better agents in the fight against cancer, two antibodies, G6 and B20-4, were isolated from synthetic antibody phage libraries. Unlike the AVASTINtrade mark antibody, a recently approved agent for the treatment of patients with colorectal cancer, B20-4 and G6 bind and block both human and murine vascular endothelial growth factor (VEGF). Here we have analyzed and compared the binding epitopes on VEGF for these three antibodies using alanine-scanning mutagenesis and structural analyses. The epitopes recognized by both synthetic antibodies are conserved between human and mouse VEGF, and they match closely to the receptor epitopes both structurally and functionally. In contrast, the Avastin epitope overlaps minimally with the receptor binding surface and centers around a residue that is not conserved in mouse. Our structural and functional analyses elucidate the cross-species reactivity of all three antibodies and emphasize the potential advantages of antibody generation using phage display as the resulting antibodies do not depend on sequence differences across species and preferentially target natural protein protein interaction surfaces. PMID- 16373346 TI - The binding of chondroitin sulfate to pleiotrophin/heparin-binding growth associated molecule is regulated by chain length and oversulfated structures. AB - Pleiotrophin is an 18-kDa heparin-binding growth factor, which uses chondroitin sulfate (CS) proteoglycan, PTPzeta as a receptor. It has been suggested that the D-type structure (GlcA(2S)beta1-3GalNAc(6S)) in CS contributes to the high affinity binding between PTPzeta and pleiotrophin. Here, we analyzed the interaction of shark cartilage CS-D with pleiotrophin using a surface plasmon resonance biosensor to reveal the importance of D-type structure. CS-D was partially digested with chondroitinase ABC, and fractionated using a Superdex 75pg column. The > or =18-mer CS fractions showed significant binding to pleiotrophin, and the longer fractions had stronger affinity for pleiotrophin than the shorter ones. The approximately 46-mer CS fraction bound to densely immobilized pleiotrophin with high affinity (K(D) = approximately 30 nM), and the binding reactions fitted the bivalent analyte model. However, when the density of the immobilized pleiotrophin was lowered, the strength of affinity remarkably decreased (K(D) = approximately 2.5 microM), and the reactions no longer fitted the model and were considered to be monovalent binding. The 20 approximately 24 mer fractions showed low affinity binding to densely immobilized pleiotrophin (K(D) = 3 approximately 20 microM), which seemed to be monovalent. When approximately 22-mer CS oligosaccharides were fractionated by strong anion exchange HPLC, each fraction differed in affinity for pleiotrophin (K(D) = 0.36 approximately >10 microM), and the affinity correlated with the amounts of D- and E- (GlcAbeta1-3GalNAc(4S,6S)) type oversulfated structures. These results suggest that the binding of pleiotrophin to CS is regulated by multivalency with CS approximately 20 mer as a unit and by the amounts of oversulfated structures. PMID- 16373347 TI - Identification and functions of chondroitin sulfate in the milieu of neural stem cells. AB - The behavior of cells is generally considered to be regulated by environmental factors, but the molecules in the milieu of neural stem cells have been little studied. We found by immunohistochemistry that chondroitin sulfate (CS) existed in the surroundings of nestin-positive cells or neural stem/progenitor cells in the rat ventricular zone of the telencephalon at embryonic day 14. Brain-specific chondroitin sulfate proteoglycans (CSPGs), including neurocan, phosphacan/receptor-type protein-tyrosine phosphatase beta, and neuroglycan C, were detected in the ventricular zone. Neurospheres formed by cells from the fetal telencephalon also expressed these CSPGs and NG2 proteoglycan. To examine the structural features and functions of CS polysaccharides in the milieu of neural stem cells, we isolated and purified CS from embryonic day 14 telencephalons. The CS preparation consisted of two fractions differing in size and extent of sulfation: small CS polysaccharides with low sulfation and large CS polysaccharides with high sulfation. Interestingly, both CS polysaccharides and commercial preparations of dermatan sulfate CS-B and an E-type of highly sulfated CS promoted the fibroblast growth factor-2-mediated proliferation of neural stem/progenitor cells. None of these CS preparations promoted the epidermal growth factor-mediated neural stem cell proliferation. These results suggest that these CSPGs are involved in the proliferation of neural stem cells as a group of cell microenvironmental factors. PMID- 16373348 TI - Mechanism of chain selection in the assembly of collagen IV: a prominent role for the alpha2 chain. AB - Collagens comprise a large superfamily of extracellular matrix proteins that play diverse roles in tissue function. The mechanism by which newly synthesized collagen chains recognize each other and assemble into specific triple-helical molecules is a fundamental question that remains unanswered. Emerging evidence suggests a role for the non-collagenous domain (NC1) located at the C-terminal end of each chain. In this study, we have investigated the molecular mechanism underlying chain selection in the assembly of collagen IV. Using surface plasmon resonance, we have determined the kinetics of interaction and assembly of the alpha1(IV) and alpha2(IV) NC1 domains. We show that the differential affinity of alpha2(IV) NC1 domain for dimer formation underlies the driving force in the mechanism of chain discrimination. Given its characteristic domain recognition and affinity for the alpha1(IV) NC1 domain, we conclude that the alpha2(IV) chain plays a regulatory role in directing chain composition in the assembly of (alpha1)(2)alpha2 triple-helical molecule. Detailed crystal structure analysis of the [(alpha1)(2)alpha2](2) NC1 hexamer and sequence alignments of the NC1 domains of all six alpha-chains from mammalian species revealed the residues involved in the molecular recognition of NC1 domains. We further identified a hypervariable region of 15 residues and a beta-hairpin structural motif of 13 residues as two prominent regions that mediate chain selection in the assembly of collagen IV. To our knowledge, this report is the first to combine kinetics and structural data to describe molecular basis for chain selection in the assembly of a collagen molecule. PMID- 16373349 TI - 3-O-sulfated oligosaccharide structures are recognized by anti-heparan sulfate antibody HS4C3. AB - Antibodies against heparan sulfate (HS) are useful tools to study the structural diversity of HS. They demonstrate the large sequential variation within HS and show the distribution of HS oligosaccharide sequences within their natural environment. We analyzed the distribution and the structural characteristics of the oligosaccharide epitope recognized by anti-HS antibody HS4C3. Biosynthetic and synthetic heparin-related oligosaccharide libraries were used in affinity chromatography, immunoprecipitation, and enzyme-linked immunosorbent assay to identify this epitope as a 3-O-sulfated motif with antithrombin binding capacity. The antibody binds weakly to any N-sulfated, 2-O- and 6-O-sulfated hexa- to octasaccharide fragment but strongly to the corresponding oligosaccharide when there is a 3-O-sulfated glucosamine residue present in the sequence. This difference was highlighted by affinity interaction and immunohistochemistry at salt concentrations from 500 mm. At physiological salt conditions the antibody strongly recognized basal lamina of epithelia and endothelia. At 500 mm salt conditions, when 3-O sulfation is required for binding, antibody recognition was more restricted and selective. Antibody HS4C3 bound similar tissue structures as antithrombin in rat kidney. Furthermore, antithrombin and antibody HS4C3 could compete with one another for binding to heparin. Antibody HS4C3 was also able to inhibit the anti-coagulant activities of heparin and Arixtra as demonstrated using the activated partial thromboplastin time clotting and the anti-factor Xa assays. In summary, antibody HS4C3 selectively detects 3-O-sulfated HS structures and interferes with the coagulation activities of heparin by association with the anti-thrombin binding pentasaccharide sequence. PMID- 16373350 TI - Structural interactions between FXYD proteins and Na+,K+-ATPase: alpha/beta/FXYD subunit stoichiometry and cross-linking. AB - Interactions of rat FXYD4 (corticosteroid hormone-induced factor (CHIF)), FXYD2 (gamma), or FXYD1 (phospholemman (PLM)) proteins with rat alpha1 subunits of Na(+),K(+)-ATPase have been analyzed by co-immunoprecipitation and covalent cross linking. In detergent-solubilized membranes from HeLa cells expressing both gamma and CHIF or CHIF and hemagglutinin A-tagged CHIF, mixed complexes of CHIF and gamma or CHIF and hemagglutinin A-tagged CHIF with alpha/beta subunits are undetectable. This implies that the alpha/beta/FXYD protomer is the major species in detergent solution. A lipid-soluble cysteine-cysteine bifunctional reagent, dibromobimane, cross-links CHIF to alpha in colonic membranes but not gamma or PLM to alpha in kidney or heart membranes, respectively. Sequence comparisons of the FXYD proteins suggested that Cys-49 in the trans-membrane segment of CHIF could be involved. In detergent-solubilized HeLa cell membranes, dibromobimane cross-links wild-type CHIF to alpha but not the C49F mutant, and also the corresponding F36C mutant but not wild-type gammab, and F48C but not wild-type PLM. C140S, C338A, C804A, and C966S mutants of the alpha subunit have been expressed. Only the C140S mutant prevents cross-linking with CHIF. The data demonstrated the proximity of trans-membrane segments of CHIF, gamma, and PLM to M2 of alpha. Molecular modeling is consistent with location of the trans-membrane segment of all FXYD proteins between M2, M6, and M9 and the proximity of Cys-49 of CHIF or Phe-36 of gamma with Cys-140 of M2. Cross-linking also demonstrated CHIF-alpha and CHIF-beta proximities in extra-membrane regions, similar to the evidence for gamma-alpha and gamma-beta cross-links. PMID- 16373351 TI - Structure of microsomal cytochrome P450 2B4 complexed with the antifungal drug bifonazole: insight into P450 conformational plasticity and membrane interaction. AB - To better understand ligand-induced structural transitions in cytochrome P450 2B4, protein-ligand interactions were investigated using a bulky inhibitor. Bifonazole, a broad spectrum antifungal agent, inhibits monooxygenase activity and induces a type II binding spectrum in 2B4dH(H226Y), a modified enzyme previously crystallized in the presence of 4-(4-chlorophenyl)imidazole (CPI). Isothermal titration calorimetry and tryptophan fluorescence quenching indicate no significant burial of protein apolar surface nor altered accessibility of Trp 121 upon bifonazole binding, in contrast to recent results with CPI. A 2.3 A crystal structure of 2B4-bifonazole reveals a novel open conformation with ligand bound in the active site, which is significantly different from either the U shaped cleft of ligand-free 2B4 or the small active site pocket of 2B4-CPI. The O shaped active site cleft of 2B4-bifonazole is widely open in the middle but narrow at the top. A bifonazole molecule occupies the bottom of the active site cleft, where helix I is bent approximately 15 degrees to accommodate the bulky ligand. The structure also defines unanticipated interactions between helix C residues and bifonazole, suggesting an important role of helix C in azole recognition by mammalian P450s. Comparison of the ligand-free 2B4 structure, the 2B4-CPI structure, and the 2B4-bifonazole structure identifies structurally plastic regions that undergo correlated conformational changes in response to ligand binding. The most plastic regions are putative membrane-binding motifs involved in substrate access or substrate binding. The results allow us to model the membrane-associated state of P450 and provide insight into how lipophilic substrates access the buried active site. PMID- 16373352 TI - Assembly of active zone precursor vesicles: obligatory trafficking of presynaptic cytomatrix proteins Bassoon and Piccolo via a trans-Golgi compartment. AB - Neurotransmitter release from presynaptic nerve terminals is restricted to specialized areas of the plasma membrane, so-called active zones. Active zones are characterized by a network of cytoplasmic scaffolding proteins involved in active zone generation and synaptic transmission. To analyze the modes of biogenesis of this cytomatrix, we asked how Bassoon and Piccolo, two prototypic active zone cytomatrix molecules, are delivered to nascent synapses. Although these proteins may be transported via vesicles, little is known about the importance of a vesicular pathway and about molecular determinants of cytomatrix molecule trafficking. We found that Bassoon and Piccolo co-localize with markers of the trans-Golgi network in cultured neurons. Impairing vesicle exit from the Golgi complex, either using brefeldin A, recombinant proteins, or a low temperature block, prevented transport of Bassoon out of the soma. Deleting a newly identified Golgi-binding region of Bassoon impaired subcellular targeting of recombinant Bassoon. Overexpressing this region to specifically block Golgi binding of the endogenous protein reduced the concentration of Bassoon at synapses. These results suggest that, during the period of bulk synaptogenesis, a primordial cytomatrix assembles in a trans-Golgi compartment. They further indicate that transport via Golgi-derived vesicles is essential for delivery of cytomatrix proteins to the synapse. Paradigmatically this establishes Golgi transit as an obligatory step for subcellular trafficking of distinct cytoplasmic scaffolding proteins. PMID- 16373353 TI - Dynamic changes in histone H3 lysine 9 methylations: identification of a mitosis specific function for dynamic methylation in chromosome congression and segregation. AB - Histone methylation is unique among post-translational histone modifications by virtue of its stability. It is thought to be a relatively stable and heritable epigenetic mark for gene-specific regulation. In this study, we use quantitative in situ approaches to investigate the cell cycle dynamics of methylated isoforms of histone H3 lysine 9. Contrary to the expected stability of trimethylated lysines, our results for trimethylated lysine 9 (tMeK9) of H3 demonstrate that the genomic content of this methylation undergoes significant changes as cells progress through mitosis. Unexpectedly, there is a loss of tMeK9 that appears to reflect a robust demethylase activity that is active during the period between anaphase and cytokinesis. Subsequent investigations of mitoses in tMeK9-deficient cells revealed defects in chromosome congression and segregation that are distinct from the increased cohesion at centromeres previously reported in association with the loss of tMeK9. Collectively, these results identify a mitosis-specific trimethylation of Lys9 in pericentromeric heterochromatin that functions in the faithful segregation of chromosomes. PMID- 16373354 TI - Yeast GTB1 encodes a subunit of glucosidase II required for glycoprotein processing in the endoplasmic reticulum. AB - Glucosidase II is essential for sequential removal of two glucose residues from N linked glycans during glycoprotein biogenesis in the endoplasmic reticulum. The enzyme is a heterodimer whose alpha-subunit contains the glycosyl hydrolase active site. The function of the beta-subunit has yet to be defined, but mutations in the human gene have been linked to an autosomal dominant form of polycystic liver disease. Here we report the identification and characterization of a Saccharomyces cerevisiae gene, GTB1, encoding a polypeptide with 21% sequence similarity to the beta-subunit of human glucosidase II. The Gtb1 protein was shown to be a soluble glycoprotein (96-102 kDa) localized to the endoplasmic reticulum lumen where it was present in a complex together with the yeast alpha subunit homologue Gls2p. Surprisingly, we found that Deltagtb1 mutant cells were specifically defective in the processing of monoglucosylated glycans. Thus, although Gls2p is sufficient for cleavage of the penultimate glucose residue, Gtb1p is essential for cleavage of the final glucose. Our data demonstrate that Gtb1p is required for normal glycoprotein biogenesis and reveal that the final two glucose-trimming steps in N-glycan processing are mechanistically distinct. PMID- 16373355 TI - Proteoglycan mechanics studied by single-molecule force spectroscopy of allotypic cell adhesion glycans. AB - Early Metazoans had to evolve the first cell adhesion system addressed to maintaining stable interactions between cells constituting different individuals. As the oldest extant multicellular animals, sponges are good candidates to have remnants of the molecules responsible for that crucial innovation. Sponge cells associate in a species-specific process through multivalent calcium-dependent interactions of carbohydrate structures on an extracellular membrane-bound proteoglycan termed aggregation factor. Single-molecule force spectroscopy studies of the mechanics of aggregation factor self-binding indicate the existence of intermolecular carbohydrate adhesion domains. A 200-kDa aggregation factor glycan (g200) involved in cell adhesion exhibits interindividual differences in size and epitope content which suggest the existence of allelic variants. We have purified two of these g200 distinct forms from two individuals of the same sponge species. Comparison of allotypic versus isotypic g200 binding forces reveals significant differences. Surface plasmon resonance measurements show that g200 self-adhesion is much stronger than its binding to other unrelated glycans such as chondroitin sulfate. This adhesive specificity through multiple carbohydrate binding domains is a type of cooperative interaction that can contribute to explain some functions of modular proteoglycans in general. From our results it can be deduced that the binding strength/surface area between two aggregation factor molecules is comparable with that of focal contacts in vertebrate cells, indicating that strong carbohydrate-based cell adhesions evolved at the very start of Metazoan history. PMID- 16373356 TI - Membrane topology of the yeast endoplasmic reticulum-localized ubiquitin ligase Doa10 and comparison with its human ortholog TEB4 (MARCH-VI). AB - Quality control machinery in the endoplasmic reticulum (ER) helps ensure that only properly folded and assembled proteins accumulate in the ER or continue along the secretory pathway. Aberrant proteins are retrotranslocated to the cytosol and degraded by the proteasome, a process called ER-associated degradation. Doa10, a transmembrane protein of the ER/nuclear envelope, is one of the primary ubiquitin ligases (E3s) participating in ER-associated degradation in Saccharomyces cerevisiae. Here we report the membrane organization of the 1319 residue Doa10 polypeptide. The topology was determined by fusing a dual-topology reporter after 16 different Doa10 fragments. Our results indicate that Doa10 contains 14 transmembrane helices (TMs). Based on protease digestion of yeast microsomes, both the N-terminal RING-CH domain and the C terminus face the cytosol. Notably, the experimentally derived topology was not predicted correctly by any of the generally available TM prediction algorithms. Bioinformatic analysis and in silico mutagenesis guided the topological studies through problematic regions. The conserved TD domain in Doa10 includes three TMs. These TMs might function in cofactor binding or substrate recognition, or they might be part of a retrotranslocation channel. The Derlins were previously proposed to provide such channels, but we show that the two yeast Derlins are not required for degradation of Doa10 membrane substrates, as was found before for the Sec61 translocon. Finally, we provide evidence that the likely human Doa10 ortholog, TEB4 (MARCH-VI), adopts a topology similar to that of Doa10. PMID- 16373358 TI - Fourth Annual International Conference on Urban Health: Achieving Social Justice in Urban Communities. Toronto, Ontario, Canada, October 26-28, 2005. Abstracts. PMID- 16373359 TI - Acute myocardial infarction and cardiac perforation on non-gated CT. PMID- 16373360 TI - Barriers to accessing benefits in a community-based insurance scheme: lessons learnt from SEWA Insurance, Gujarat. AB - This paper seeks to examine barriers faced by members of a community-based insurance (CBI) scheme, which is targeted at poor women and their families, in accessing scheme benefits. CBI schemes have been developed and promoted as mechanisms to offer protection to poor families from the risks of ill-health, death and loss of assets. However, having voluntarily enrolled in a CBI scheme, poor households may find it difficult or impossible to access scheme benefits. The paper describes the results of qualitative research carried out to assess the barriers faced in accessing scheme benefits by members of the CBI scheme run by the Self-Employed Women's Association (SEWA) in Gujarat, India. The study finds that the members face a variety of different barriers, particularly in seeking hospitalization and in submitting insurance claims. Some of the barriers are rooted in factors outside the scheme's control, such as illiteracy and financial poverty amongst members, and inadequacies of the transportation and health care infrastructure. But other barriers relate to the scheme's design and management, for example, lack of clarity among scheme staff regarding the scheme's rules and processes, and requirements that claimants submit documents to prove the validity of their claims. The paper makes recommendations as to how SEWA Insurance can address some of the identified barriers and discusses the relevance of these findings to other CBI schemes in India and elsewhere. PMID- 16373361 TI - Lipoprotein is a predominant Toll-like receptor 2 ligand in Staphylococcus aureus cell wall components. AB - Lipoteichoic acid (LTA) derived from Staphylococcus aureus is reported to be a ligand of Toll-like receptor 2 (TLR2). In this study, we demonstrated that lipoproteins obtained from S. aureus are potent activators of TLR2. A fraction obtained by Triton X-114 phase partitioning activated cells through TLR2. The fraction contained proteins and LTA. The activity was detected in compounds in a mass range of 12-40 kDa. Proteinase K digested the active compounds into lower molecular weight active materials <10 kDa. In contrast, hydrofluoric acid treatment, which decomposes LTA, did not alter the molecular mass of the active compounds. Further, most of the activity was abrogated by lipoprotein lipase digestion. These results suggested that lipoproteins are predominant TLR2 ligands in S. aureus cell wall components. PMID- 16373362 TI - Differential expression of suppressors of cytokine signaling-1 and -3 and related cytokines in central nervous system during remitting versus non-remitting forms of experimental autoimmune encephalomyelitis. AB - SJL mice exhibit a relapsing-remitting course of experimental autoimmune encephalomyelitis (EAE), whereas C57BL/6 (B6) mice display a more chronic course without complete remissions. Suppressor of cytokine signaling (SOCS)-1 and SOCS-3 are members of a family of inducible intracellular proteins that negatively regulate cytokine signaling in cells of hematopoietic origin and may influence the Th1 to Th2 balance. SOCS-1 and SOCS-3 are induced by cytokines that are known to be up-regulated during EAE, including IFN-gamma (IFN-g) and IL-6, respectively. To test the hypothesis that the level of induction of SOCS-1 and SOCS-3 correlates with the course of EAE, mRNA levels were compared in spinal cords of SJL and B6 mice during discrete stages of disease. SOCS-1 and SOCS-3 were elevated throughout active disease in both strains. At peak EAE, SOCS-1 was higher and SOCS-3 was lower in B6 cords compared with SJL cords. This correlated with greater expression of the Th1 cytokine, IFN-g, and less of the Th2 cytokine, IL-10, in B6 cords relative to SJL cords during onset and peak disease. SOCS-3 inducers in the IL-6 family were expressed differentially between the strains. IL 6 and leukemia inhibitory factor were higher at onset in B6 cords whereas ciliary neurotrophic factor was increased in SJL cords during peak disease. Expression of fibroblast growth factor-2, which may be involved in remyelination, was higher in SJL cords at peak. Comparison of these models suggests that cytokine autoregulatory mechanisms involving SOCS may play a role in determining the course of EAE. PMID- 16373363 TI - Combined CD137 (4-1BB) and adjuvant therapy generates a developing pool of peptide-specific CD8 memory T cells. AB - In practice, vaccines should induce lasting and efficacious T cell immunity without promoting deleterious pathological consequences. To accomplish this goal we immunized mice with ovalbumin peptide, polyinosinic-polycytidylic and anti CD137. Vaccinated mice retained a massive functional CD8 T cell memory pool in lymphoid and non-lymphoid tissues for >1 year. The memory T cells clonally expanded, produced substantial amounts of IFNgamma, and responded vigorously to vesicular stomatitis virus infection. To understand how the vaccine might function, we showed that the antigen-specific T cells must bear CD137 in order for optimal priming to occur. Thus, anti-CD137 agonist mAb directly stimulated peptide-specific CD8 T cells and conditioned them to survive. In contrast, CD137 deficient CD8 T cells did not survive despite CD137 expression by antigen presenting cells. Taken together, the data indicate that CD137 and adjuvant combined therapy is an efficacious vaccine strategy for immunization with non replicating inert antigen. PMID- 16373364 TI - A role for Ets1, synergizing with AP-1 and GATA-3 in the regulation of IL-5 transcription in mouse Th2 lymphocytes. AB - IL-5 is a key regulator of eosinophilic inflammation and is selectively expressed by antigen-activated Th2 lymphocytes. An important role for the proximal AP-1 and GATA sites in regulating IL-5 transcription is generally accepted but the significance of an adjacent Ets/NFAT site has remained unclear. We have investigated its role using the mouse Th2 clone D10.G4.1. Transcription of IL-5 reporter gene plasmids could be induced in D10 cells by phorbol myristate acetate/cyclic adenosine monophosphate (PMA/cAMP) stimulation and significantly further enhanced by activation of the mitogen-activated protein (MAP) kinase pathways. Strong induction of IL-5 mRNA was also induced by PMA/cAMP. Mutagenesis showed that the Ets/NFAT site is of critical importance along with the AP-1 and GATA sites in regulating IL-5 transcription stimulated by PMA/cAMP and MAP kinase activation. Transactivation was used to investigate the transcription factors which could function at the three sites and possible synergistic interactions. AP 1 (c-Fos/c-Jun) strongly induced IL-5 transcription and dominant negative AP-1 constructs confirmed that AP-1 plays an important role in regulating IL-5 expression. Ets1, unlike other members of the Ets/NFAT family, synergized strongly with AP-1 suggesting that Ets1 is the family member which functions at the Ets/NFAT site. AP-1/Ets1 transactivation also stimulated IL-5 mRNA expression. Ets1 binding to the proximal promoter region, demonstrated by chromatin immunoprecipitation, was stimulated by PMA/cAMP. The absolute dependence on the binding sites for Ets1, AP-1 and GATA-3 together with the strong synergy between Ets1 and AP-1 suggest close cooperative interactions between the three transcription factors in the regulation of IL-5 expression in mouse T cells. PMID- 16373365 TI - Lymphoid B cells induce NF-kappaB activation in high endothelial cells from human tonsils. AB - Immune surveillance depends on still poorly understood lymphocyte-endothelium interactions required for lymphocyte transendothelial migration into secondary lymphoid organs. The nuclear factor kappaB (NF-kappaB) regulatory system and its inhibitory IkappaB proteins control the inducible expression of adhesion molecules, cytokines and chemokines involved in endothelial activation and lymphocyte transmigration. Here we present results showing the activation of this system in response to the interaction of high endothelial cells from human tonsils (HUTEC) with human B and T lymphoid cell lines and primary tonsillar lymphocytes. Western blot and electrophoretic mobility shift assays show that adhesion of different lymphoid cells induce varying levels of NF-kappaB activation in HUTEC, with Daudi cells, tonsil-derived B cell line 10 (TBCL-10) and primary tonsillar B lymphocytes causing the strongest activation. The main NF kappaB protein complexes translocated to the nucleus were p65/p50 and p50/p50. Results from reverse transcription-PCR and flow cytometry analysis of HUTEC indicate that the interaction with Daudi cells induce an increased expression of IL-6 and IL-8 mRNA and cell-surface expression of intercellular adhesion molecule 1, all of which were prevented by sodium salicylate, an inhibitor of NF-kappaB activation. Transwell experiments show that NF-kappaB activation and the response of HUTEC to the interaction of Daudi cells does not depend on direct cell-cell contact but rather on the production of soluble factors that require the presence of both cell types. These results suggest that lymphocytes and high endothelium establish a cross talk leading to NF-kappaB-mediated expression of cytokines and adhesion molecules, inducing endothelial cell activation. PMID- 16373366 TI - Maternal and offspring MTHFR gene C677T polymorphism as predictors of congenital atrial septal defect and patent ductus arteriosus. AB - To observe the association of MTHFR gene C677T locus polymorphism with occurrence of congenital heart defects (CHDs), 21 patients with atrial septal defect (ASD), 35 patients with patent ductus arteriosus (PDA), one patient with both conditions combined, and their biological parents were collected as the case group. Another 104 normal individuals and their biological parents without a family history of birth defects were selected as the control group. MTHFR C677T genotypes of each sample were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The results showed for the occurrence of ASD, the odds ratio (OR) of TT genotype was 4.08 [95% confidence interval (95% CI) = 1.28 13.24] compared with CT genotype. For the occurrence of PDA, the ORs of TT were 3.44 (95% CI = 0.89-16.13) and 2.38 (95% CI = 0.92-6.14) compared with CC and CT genotypes, respectively. Author as meant? Compared with CC + CT genotype combination, the ORs of TT were 3.95 (95% CI = 1.38-11.44) and 2.60 (95% CI = 1.02-6.36) for ASD and PSD respectively. The results also had sex differences and the statistical significance was only observed in male ASD and female PDA. The ORs of T allele carriers were 2.29 (95% CI = 1.08-4.92) and 1.88 (95% CI = 1.02 3.47) compared with C allele for the occurrences of ASD and PDA respectively. The analysis of parents genotype showed that the OR of TT mothers was 2.31 (95% CI = 0.96-5.59, P < 0.05) compared with (CC + CT) for the occurrence of PDA in offspring. So this study could give a clue that MTHFR C677T locus variation was related with occurrence of ASD and PDA, and the carriers of TT genotype and T allele had higher risk of diseases. The mother carrying TT genotype was associated with occurrence of PDA in offspring. PMID- 16373367 TI - Mitochondrial DNA deletions in rhesus macaque oocytes and embryos. AB - Mitochondria are the most abundant organelles in mammalian oocytes and early embryos. Mitochondrial DNA (mtDNA) mutations, including the common deletion, have been found in skeletal muscle fibres from aged rhesus macaques. The specific aims of this study were to determine whether the mitochondrial common deletion is present in rhesus oocytes after hormonal stimulation and in embryos generated by in vitro production, or whether this deletion is already present in the immature oocyte. Using a nested primer PCR strategy, we found a significant increase in the proportion of mtDNA deletions in stimulated oocytes and embryos from rhesus macaques, compared with mtDNA deletions in immature, unstimulated oocytes derived from necropsied ovaries of age-matched monkeys. The common deletion is larger in the rhesus (5704 bp) than in humans (4977 bp). Accumulation of mtDNA deletions in oocytes may contribute to mitochondrial dysfunction and impaired ATP production. We propose the rhesus to be an excellent model to assess the quality of gametes and embryos and their developmental competence in primates, including humans. PMID- 16373368 TI - Variants of the CTLA4 gene that segregate with autoimmune diseases are not associated with endometriosis. AB - An autoimmune etiology has been suggested for endometriosis mostly on the basis of an increased prevalence of autoimmune diseases in affected women. Cytotoxic T lymphocyte antigen (CTLA) 4 gene is recognized as a primary determinant for autoimmunity since specific polymorphisms have been associated with predisposition to most autoimmune disorders. This study was aimed to evaluate whether two variants of CTLA4 gene might be associated with endometriosis in an Italian population. We examined the +49A/G polymorphism and the CT60A/G dimorphism in n = 146 endometriosis subjects classified according to Holt and Weiss criteria. Controls were represented by n = 165 women without laparoscopic evidence of the disease. We found no statistically significant difference in the genotype frequencies between women with and without endometriosis. The proportion of the mutant G allele of the +49A/G polymorphism in the former and in the latter group resulted 34 and 30%, respectively. The proportion of the susceptible G allele of the CT60 A/G dimorphism resulted 51% in both groups. No association was demonstrated between the polymorphisms and the clinical forms of the disease and no susceptibility haplotypes were found. These findings suggest that endometriosis aetiology is not primarily associated with the development of CTLA4 linked autoimmunity. PMID- 16373369 TI - Influence of inorganic nitrogen and pH on the elongation of maize seminal roots. AB - BACKGROUND AND AIMS: Root absorption and assimilation of inorganic nitrogen usually alters rhizosphere pH, but the immediate influence of such pH changes on root elongation as well as that of exogenous inorganic nitrogen itself has been uncertain. METHODS: A differential extensiometer that monitored on a real-time, continuous basis root elongation in an intact 3-d-old maize plant was developed. Treatments included root media at pH 6.5 or 5.6 that lacked nitrogen and ones at pH 6.5 that contained 100 mmol m(-3) NH(4)(+) or NO(3)(-). KEY RESULTS: Acidifying the root medium from pH 6.5 to 5.6 nearly doubled the elasticity of the seminal root, but slightly decreased its elongation. Plasticity of the root apex was not detectable in all treatments. The presence of ammonium or nitrate in the medium stimulated elongation by 29 % or 14 %, respectively. Addition of an osmoticum to the medium had no effect on root elongation in the absence of inorganic nitrogen, but diminished the stimulation of elongation in the presence of ammonium and nitrate. This indicates that these ions or their by-products serve partially as osmolytes. CONCLUSIONS: In nutrient solution, root elongation of a maize seedling--even one with ample nitrogen reserves--depended most strongly on exogenous inorganic nitrogen, and less so, if at all, on either the pH of the bulk nutrient solution or the mechanical properties of cell walls. PMID- 16373370 TI - Nectar concentration and composition of 26 species from the temperate forest of South America. AB - BACKGROUND AND AIMS: Floral nectar concentration and chemical composition of 26 plant species native to the temperate forest of southern South America are reported and the relationships with the flower type are evaluated. METHODS: Nectar concentration was measured with a hand refractometer and sugar composition was analysed by gas-liquid chromatography. Plant species were classified into flower type categories based not only on floral features but also on data from the literature and field observations on their pollinators. KEY RESULTS: Most data on nectar are new reports at the generic and/or specific level. Plant species in which more than one population was studied showed significant among population variation in nectar sugar concentration and composition. Results showed a weak relationship between nectar traits and flower type. Many species had nectar containing 50 % or more sucrose (17 of 26 species), independent of the main pollinator. CONCLUSIONS: Considering that (a) nectar characteristics did not show a clear association with different flower types or with plant taxonomic membership, and (b) different populations of the same species showed large variability in sugar composition, the results suggest that other factors (e.g. historical and environmental) could be involved in determining the sugar composition of the highly endemic plant species from this region. PMID- 16373371 TI - Anther cap retention prevents self-pollination by elaterid beetles in the South African orchid Eulophia foliosa. AB - BACKGROUND AND AIMS: Pollination by insects that spend long periods visiting many flowers on a plant may impose a higher risk of facilitated self-pollination. Orchids and asclepiads are particularly at risk as their pollen is packaged as pollinia and so can be deposited on self-stigmas en masse. Many orchids and asclepiads have adaptations to limit self-deposition of pollinia, including gradual reconfiguration of pollinaria following removal. Here an unusual mechanism--anther cap retention--that appears to prevent self-pollination in the South African orchid Eulophia foliosa is examined. METHODS: Visits to inflorescences in the field were observed and pollinators collected. Visitation rates to transplanted inflorescences were compared between a site where putative pollinators were abundant and a site where they were rare. Anther cap retention times were determined for removed pollinaria and atmospheric vapour pressure deficit was recorded concurrently. Anther cap anatomy was examined using light microscopy. KEY RESULTS: Eulophia foliosa is pollinated almost exclusively by Cardiophorus obliquemaculatus (Elateridae) beetles, which remain on the deceptive inflorescences for on average 301 s (n = 18). The anther cap that covers the pollinarium is retained for an average of 512 s (n = 24) after pollinarium removal by beetles. In all populations measured, anther cap dimensions are greater than those of the stigmatic cavity, thus precluding the deposition of self-pollinia until after the anther cap has dropped. An anatomical investigation of this mechanism suggests that differential water loss from regions of the anther cap results in opening of the anther cap flaps. This is supported by observations that as atmospheric vapour pressure deficits increased, the duration of anther cap retention was reduced. CONCLUSIONS: Flowers of E. foliosa are specialized for pollination by elaterid beetles. Retention of anther caps for a period exceeding average visit times by beetles to inflorescences appears to prevent facilitated self-pollination in E. foliosa effectively. PMID- 16373372 TI - Physical activity and change in body mass index from adolescence to mid-adulthood in the 1958 British cohort. AB - BACKGROUND: Prevention of obesity has focused on childhood as a target period. Our aim was to assess whether frequency of adolescent physical activity affected subsequent body mass index (BMI) gain through to mid-adulthood. METHODS: The British birth cohort of all births in 1 week in March 1958, includes information on physical activity frequency and BMI for several ages, 11-45 years. We examined relationships between activity in adolescence and trajectories of BMI between 16 years (or 23 years) and 45 years using multi-level models. Effects of change in activity on BMI and on change in BMI were tested using ANOVA. RESULTS: Physical activity at 11 years had no effect on the BMI trajectories, in males or females. More active females at 16 years gained BMI more slowly than others, by 0.007 kg/m2/year per activity category over the period 16-45 years, whereas the most active males gained BMI faster than others, by 0.005 kg/m2/year per activity category. This effect in males was not evident on the BMI trajectory from 23 to 45 years. Consistent with these analyses, change in activity was associated with change in BMI in females, e.g. females active at 16 and 42 years gained less BMI than inactive females (2.1 vs 2.5 kg/m2/10 years). Results for males were inconsistent over the time periods examined. CONCLUSIONS: Physical activity may lessen the gains in BMI from adolescence onwards, but relationships vary with age, and in later adolescence show opposite effects for males and females. Decreasing activity between adolescence and mid-adulthood in males, and inactivity in both life stages in females may increase BMI gain. PMID- 16373374 TI - Genes underlying common complex diseases. PMID- 16373375 TI - Cohort profile: the 1982 Pelotas (Brazil) birth cohort study. PMID- 16373376 TI - Mother's consent to linkage of survey data with her child's birth records in a multi-ethnic national cohort study. AB - BACKGROUND: The increased use of computer-based records has facilitated linkage of routine data with that obtained for research. When children are involved, parental consent for linkage is usually required. The Millennium Cohort Study, of 18,819 UK babies born in 2000-02, over-sampled families from disadvantaged and ethnic wards, providing the opportunity to investigate factors associated with mother's consent to access her child's birth records. METHODS: Factors considered included ward type and mother's socioeconomic status, ethnicity, education, age, and language. Logistic regression was used to investigate the relationship of these factors with consent. RESULTS: Consent for linkage to birth register and/or hospital maternity data was obtained from 92% of the cohort mothers. The proportions consenting differed according to the mother's country of residence, age, and education, with consent being less likely among minority ethnic group mothers, lone parents, and those with higher degrees or no qualifications. Where interviews had been translated, consent was significantly less likely if the interpreter was a male. CONCLUSION: A large proportion of mothers who were interviewed gave permission for linkage. However, there were some groups who were less likely to do so, particularly those from minority ethnic groups. These sources of non-consent bias should be taken into account when analysing linked data from socially and ethnically mixed populations. Efforts should be made to understand the reasons for non-consent, which in turn will help determine the best ways to encourage more mothers to consent in future. PMID- 16373377 TI - Occupational exposures and risks of liver cancer among Shanghai female textile workers--a case-cohort study. AB - BACKGROUND: Liver cancer is the fifth most frequent malignancy worldwide. Viral hepatitis B and C, alcohol, and aflatoxin are the major established risk factors. Little is known about the aetiological contributions of occupational exposures, as previous occupational epidemiological studies of liver cancer suggest few agent-specific associations. We investigated associations of occupational exposures to dusts and chemicals in a cohort of female textile workers. METHODS: Cancer incidence was determined among 267,400 female textile workers in Shanghai, China, who had been enrolled in an intervention trial of breast self-exam efficacy during 1989-98. Subjects were interviewed at baseline regarding basic demographics, smoking habits, alcohol consumption, and contraceptive practices. A case-cohort study of 360 liver cancer cases and 3,186 age-stratified randomly chosen subcohort subjects was conducted within this cohort. Exposures to workplace dusts and chemicals were reconstructed from complete work history data, historical exposure monitoring data for selected agents, and a specially designed job-exposure matrix for the textile industry. Relative risks and dose-response trends were estimated by Cox proportional hazards modelling, adapted for the case cohort design. Latency analyses with different lag years were also applied. RESULTS: 2,095,904 person-years were contributed by this female cohort. The results of the case-cohort analysis revealed a protective effect of cotton fibre exposure years [adjusted hazards ratio (HR) = 0.64; 95% confidence interval (95% CI) 0.44-0.92] or endotoxin exposure (adjusted HR = 0.60; 95% CI 0.41-0.88) for the fourth quartile with significant trends for 20 year exposure lags. CONCLUSIONS: This study suggests that chronic exposure to endotoxin or some other component of cotton dust exposure may have reduced liver cancer risk in this population. PMID- 16373378 TI - Weight change and changes in the metabolic syndrome as the French population moves towards overweight: the D.E.S.I.R. cohort. AB - BACKGROUND: How weight change affects the metabolic syndrome (MS) and its parameters is unknown, particularly, in a leaner European population such as the French prospective D.E.S.I.R. cohort. METHODS: In 3770 D.E.S.I.R. participants (sex ratio=1) averaging 47.5 years (range 30-64), with measured weight and MS parameters at baseline (D0) and at 6 year follow-up (D6), we assessed this relationship across five weight-change classes, using stable weight as the referent group (-2 to +2 kg). We used analysis-of-covariance to assess changes in each MS parameter and logistic regression to assess incident MS, according to the National Cholesterol Education Program (NCEP). We also assessed weight-change effect on MS status between D0 and D6. RESULTS: At D0, average weight was 68.4 kg (SD 12.3); BMI was 24.8 kg/m2 (SD 3.5). From D0-D6, the cohort gained a mean 2.1 kg (median 2.0; SD 4.4). After adjustment for age and D0 weight, there was a strong linear relationship with weight change and worsening of the following MS parameters at D6: fasting insulin, waist girth, fasting glucose, fasting triglycerides, HDL cholesterol, and systolic and diastolic blood pressure (P<0.0001). After age adjustment, for every kilogram gained over 6 years, risk of developing the NCEP Syndrome increased 22% (OR 1.22; 95% CI 1.18-1.25). NCEP-MS was incident in 3% of those with stable weight compared with 21% among those gaining >9 kg; 10% of those who lost >2 kg reverted to non-NCEP-MS. CONCLUSIONS: All continuous MS measures are linearly related to weight change, and MS can resolve with modest weight loss, underscoring the importance of maintaining lifelong normal weight. PMID- 16373379 TI - Healthy nature healthy people: 'contact with nature' as an upstream health promotion intervention for populations. AB - Whilst urban-dwelling individuals who seek out parks and gardens appear to intuitively understand the personal health and well-being benefits arising from 'contact with nature', public health strategies are yet to maximize the untapped resource nature provides, including the benefits of nature contact as an upstream health promotion intervention for populations. This paper presents a summary of empirical, theoretical and anecdotal evidence drawn from a literature review of the human health benefits of contact with nature. Initial findings indicate that nature plays a vital role in human health and well-being, and that parks and nature reserves play a significant role by providing access to nature for individuals. Implications suggest contact with nature may provide an effective population-wide strategy in prevention of mental ill health, with potential application for sub-populations, communities and individuals at higher risk of ill health. Recommendations include further investigation of 'contact with nature' in population health, and examination of the benefits of nature-based interventions. To maximize use of 'contact with nature' in the health promotion of populations, collaborative strategies between researchers and primary health, social services, urban planning and environmental management sectors are required. This approach offers not only an augmentation of existing health promotion and prevention activities, but provides the basis for a socio ecological approach to public health that incorporates environmental sustainability. PMID- 16373380 TI - Enhancing vacuolar sucrose cleavage within the developing potato tuber has only minor effects on metabolism. AB - Modification of tuber carbohydrate metabolism by the tuber-specific expression of a yeast invertase targeted to the cytosol or apoplast has previously been demonstrated to have diverse effects on tuber growth and metabolism. In the current study, we generated plants exhibiting tuber-specific expression of the same enzyme targeted to the vacuole. Enzymatic analysis of the carbohydrate levels of the tuber revealed dramatic decreases in sucrose content coupled with large increases in the levels of glucose and hexose phosphates, but unaltered starch content in the transformants. Analysis of the key enzyme of glycolysis suggests that this pathway is down-regulated in the transformants. Despite these changes in metabolite pools and enzyme activity, few consistent changes could be observed in the estimated metabolic fluxes following incubation of isolated tuber discs in labelled glucose. The analysis of the relative levels of a wide range of metabolites using a gas chromatography-mass spectrometry (GC-MS)-based metabolite profiling method revealed large changes in the levels of fructose and decreases in a range of other sugars, but very few changes in the contents of organic and amino acids. This metabolic profile is remarkably consistent with that obtained following expression of the invertase in the apoplastic compartment, providing circumstantial evidence for the endocytotic trafficking of sugars within potato tuber parenchyma. Finally, the results of this study are compared with those from other plant species and the relative roles of the vacuolar isoform of the enzyme are contrasted. PMID- 16373381 TI - In vivo fragmentation of the large subunit of ribulose-1,5-bisphosphate carboxylase by reactive oxygen species in an intact leaf of cucumber under chilling-light conditions. AB - Previous studies have demonstrated that the large subunit (LSU) of ribulose-1,5 bisphosphate carboxylase (Rubisco) is site-specifically cleaved by a hydroxyl radical (*OH) generated in the illuminated chloroplast lysates or by an artificial *OH-generating system. However, it is not known whether such cleavage of the LSU by reactive oxygen species (ROS) actually occurs in an intact leaf. When leaf discs of chilling-sensitive cucumber (Cucumis sativus L.) were illuminated at 4 degrees C, five major fragments of the LSU were observed. This fragmentation was completely inhibited by ROS scavengers, such as n-propyl gallate (for *OH) and 1,2-dihydroxybenzene-3,5-disulfonic acid (Tiron) (for superoxide). FeSO4 stimulated this fragmentation, whereas an iron-specific chelator, deferoxamine, suppressed it. Furthermore, such fragments were identical to those generated from the purified Rubisco by an *OH-generating system in vitro on two-dimensional PAGE. These results indicate that the direct fragmentation of the LSU by reactive oxygen species also occurs in an intact leaf. PMID- 16373382 TI - Toward a unified hypothesis of interneuronal modulation. PMID- 16373383 TI - ATP-sensitive K+ channels: regulation of bursting by the sulphonylurea receptor, PIP2 and regions of Kir6.2. AB - ATP-sensitive K+ channels composed of the pore-forming protein Kir6.2 and the sulphonylurea receptor SUR1 are inhibited by ATP and activated by Phosphatidylinositol Bisphosphate (PIP2). Residues involved in binding of these ligands to the Kir6.2 cytoplasmic domain have been identified, and it has been hypothesized that gating mechanisms involve conformational changes in the regions of the bundle crossing and/or the selectivity filter of Kir6.2. Regulation of Kir6.2 by SUR1, however, is not well-understood, even though this process is ATP and PIP2 dependent. In this study, we investigated the relationship between channel regulation by SUR1 and PIP2 by comparing a number of single and double mutants known to affect open probability (P(o)), PIP2 affinity, and sulphonylurea and MgADP sensitivity. When coexpressed with SUR1, the Kir6.2 mutant C166A, which is characterized by a P(o) value close to 0.8, exhibits no sulphonylurea or MgADP sensitivity. However, when P(o) was reduced by combining mutations at the PIP2 sensitive residues R176 and R177 with C166A, sulphonylurea and MgADP sensitivities were restored. These effects correlated with a dramatic decrease in PIP2 affinity, as assessed by PIP2-induced channel reactivation and inhibition by neomycin, an antagonist of PIP2 binding. Based on macroscopic and single-channel data, we propose a model in which entry into the high-P(o) bursting state by the C166A mutation or by SUR1 depends on the interaction of PIP2 with R176 and R177 and, to a lesser extent, R54. In conjunction with this PIP2-dependent process, SUR1 also regulates channel activity via a PIP2-independent, but MgADP-dependent process. PMID- 16373384 TI - Effect of inspiratory muscle work on peripheral fatigue of locomotor muscles in healthy humans. AB - The work of breathing required during maximal exercise compromises blood flow to limb locomotor muscles and reduces exercise performance. We asked if force output of the inspiratory muscles affected exercise-induced peripheral fatigue of locomotor muscles. Eight male cyclists exercised at > or = 90% peak O2 uptake to exhaustion (CTRL). On a separate occasion, subjects exercised for the same duration and power output as CTRL (13.2 +/- 0.9 min, 292 W), but force output of the inspiratory muscles was reduced (-56% versus CTRL) using a proportional assist ventilator (PAV). Subjects also exercised to exhaustion (7.9 +/- 0.6 min, 292 W) while force output of the inspiratory muscles was increased (+80%versus CTRL) via inspiratory resistive loads (IRLs), and again for the same duration and power output with breathing unimpeded (IRL-CTRL). Quadriceps twitch force (Q(tw)), in response to supramaximal paired magnetic stimuli of the femoral nerve (1-100 Hz), was assessed pre- and at 2.5 through to 70 min postexercise. Immediately after CTRL exercise, Q(tw) was reduced -28 +/- 5% below pre-exercise baseline and this reduction was attenuated following PAV exercise (-20 +/- 5%; P < 0.05). Conversely, increasing the force output of the inspiratory muscles (IRL) exacerbated exercise-induced quadriceps muscle fatigue (Q(tw) = -12 +/- 8% IRL CTRL versus-20 +/- 7% IRL; P < 0.05). Repeat studies between days showed that the effects of exercise per se, and of superimposed inspiratory muscle loading on quadriceps fatigue were highly reproducible. In conclusion, peripheral fatigue of locomotor muscles resulting from high-intensity sustained exercise is, in part, due to the accompanying high levels of respiratory muscle work. PMID- 16373385 TI - Topographic organization in the auditory brainstem of juvenile mice is disrupted in congenital deafness. AB - There is an orderly topographic arrangement of neurones within auditory brainstem nuclei based on sound frequency. Previous immunolabelling studies in the medial nucleus of the trapezoid body (MNTB) have suggested that there may be gradients of voltage-gated currents underlying this tonotopic arrangement. Here, our electrophysiological and immunolabelling results demonstrate that underlying the tonotopic organization of the MNTB is a combination of medio-lateral gradients of low-and high-threshold potassium currents and hyperpolarization-activated cation currents. Our results also show that the intrinsic membrane properties of MNTB neurones produce a topographic gradient of time delays, which may be relevant to sound localization, following previous demonstrations of the importance of the timing of inhibitory input from the MNTB to the medial superior olive (MSO). Most importantly, we demonstrate that, in the MNTB of congenitally deaf mice, which exhibit no spontaneous auditory nerve activity, the normal tonotopic gradients of neuronal properties are absent. Our results suggest an underlying mechanism for the observed topographic gradient of neuronal firing properties in the MNTB, show that an intrinsic neuronal mechanism is responsible for generating a topographic gradient of time-delays, and provide direct evidence that these gradients rely on spontaneous auditory nerve activity during development. PMID- 16373386 TI - Differential polyamine sensitivity in inwardly rectifying Kir2 potassium channels. AB - Recent studies have shown that Kir2 channels display differential sensitivity to intracellular polyamines, and have raised a number of questions about several properties of inward rectification important to the understanding of their physiological roles. In this study, we have carried out a detailed characterization of steady-state and kinetic properties of block of Kir2.1-3 channels by spermine. High-resolution recordings from outside-out patches showed that in all Kir2 channels current-voltage relationships display a 'crossover' effect upon change in extracellular K+. Experiments at different concentrations of spermine allowed for the characterization of two distinct shallow components of rectification, with the voltages for half-block negative (V1(1/2)) and positive (V2(1/2)) to the voltage of half-block for the major steep component of rectification (V0(1/2)). While V1(1/2) and V2(1/2) voltages differ significantly between Kir2 channels, they were coupled to each other according to the equation V1(1/2)-V2(1/2) = constant, strongly suggesting that similar structures may underlie both components. In Kir2.3 channels, the V2(1/2) was approximately 50 mV positive to V0(1/2), leading to a pattern of outward currents distinct from that of Kir2.1 and Kir2.2 channels. The effective valency of spermine block (Z0) was highest in Kir2.2 channels while the valencies in Kir2.1 and Kir2.3 channels were not significantly different. The voltage dependence of spermine unblock was similar in all Kir2 channels, but the rates of unblock were approximately 7-fold and approximately 16-fold slower in Kir2.3 channels than those in Kir2.1 and Kir2.2 when measured at high and physiological extracellular K+, respectively. In all Kir2 channels, the instantaneous phase of activation was present. The instantaneous phase was difficult to resolve at high extracellular K+ but it became evident and accounted for nearly 30-50% of the total current when recorded at physiological extracellular K+. In conclusion, the data are consistent with the universal mechanism of rectification in Kir2 channels, but also point to significant, and physiologically important, quantitative differences between Kir2 isoforms. PMID- 16373388 TI - Bone morphogenic protein-7 and the kidney: current concepts and open questions. PMID- 16373387 TI - Expression and biophysical properties of Kv1 channels in supragranular neocortical pyramidal neurones. AB - Potassium channels are extremely diverse regulators of neuronal excitability. As part of an investigation into how this molecular diversity is utilized by neurones, we examined the expression and biophysical properties of native Kv1 channels in layer II/III pyramidal neurones from somatosensory and motor cortex. Single-cell RT-PCR, immunocytochemistry, and whole cell recordings with specific peptide toxins revealed that individual pyramidal cells express multiple Kv1 alpha-subunits. The most abundant subunit mRNAs were Kv1.1 > 1.2 > 1.4 > 1.3. All of these subunits were localized to somatodendritic as well as axonal cell compartments. These data suggest variability in the subunit complexion of Kv1 channels in these cells. The alpha-dendrotoxin (alpha-DTX)-sensitive current activated more rapidly and at more negative potentials than the alpha-DTX insensitive current, was first observed at voltages near action potential threshold, and was relatively insensitive to holding potential. The alpha-DTX sensitive current comprised about 10% of outward current at steady-state, in response to steps from -70 mV. From -50 mV, this percentage increased to approximately 20%. All cells expressed an alpha-DTX-sensitive current with slow inactivation kinetics. In some cells a transient component was also present. Deactivation kinetics were voltage dependent, such that deactivation was slow at potentials traversed by interspike intervals during repetitive firing. Because of its kinetics and voltage dependence, the alpha-DTX-sensitive current should be most important at physiological resting potentials and in response to brief stimuli. Kv1 channels should also be important at voltages near threshold and corresponding to interspike intervals. PMID- 16373389 TI - Long-term lamivudine therapy is not reasonable for HBV-associated nephropathy. PMID- 16373390 TI - Recurrent paroxysms of metabolic acidosis in a haemodialysis patient. PMID- 16373391 TI - Phenolphthalein and bisacodyl: assessment of genotoxic and carcinogenic responses in heterozygous p53 (+/-) mice and syrian hamster embryo (SHE) assay. AB - Phenolphthalein (800 and 2400 mg/kg/day by gavage and 2400 mg/kg/day by diet) and bisacodyl (800-500, 4000-2000, and 8000 mg/kg/day by gavage) were administered to 15 male and 15 female and 20 male and 20 female p53(+/-) mice respectively for 26 weeks to investigate the potential carcinogenicity of each compound. Toxicokinetic analyses confirmed systemic exposure. p-Cresidine was administered by gavage (400 mg/kg/day) and served as the positive control agent in each study. Dietary phenolphthalein reduced survival in both sexes and early deaths were attributed to thymic lymphoma. No bisacodyl-related neoplasms were observed. Regardless of route of administration to p53(+/-) mice, phenolphthalein but not bisacodyl was unequivocally genotoxic, causing increased micronuclei in polychromatic erythrocytes. In the Syrian hamster embryo (SHE) cell transformation assay, phenolphthalein caused increases in morphologically transformed colonies, thereby corroborating NTP's earlier reports, showing phenolophthalein has potential carcinogenic activity. Bisacodyl was negative in the SHE assay. Results of these experiments confirm an earlier demonstration that dietary phenolphthalein causes thymic lymphoma in p53(+/-) mice and show that (1) phenolphthalein causes qualitatively identical results in this transgenic model regardless of route of oral administration, (2) phenolphthalein shows evidence of micronucleus induction in p53(+/-) mice for up to 26 weeks, (3) phenolphthalein induced transformations in the in vitro SHE assay, and (4) bisacodyl in p53(+/-) mice induces neither drug-related neoplasm, nor micronuclei in polychromatic erythrocytes, and did not induce transformations in the in vitro SHE assay. PMID- 16373392 TI - Periodic extinctions of transposable elements in bacterial lineages: evidence from intragenomic variation in multiple genomes. AB - Most previous work on the evolution of mobile DNA was limited by incomplete sequence information. Whole genome sequences allow us to overcome this limitation. I study the nucleotide diversity of prominent members of five insertion sequence families whose transposition activity is encoded by a single transposase gene. Eighteen among 376 completely sequenced bacterial genomes and plasmids carry between 3 and 20 copies of a given insertion sequence. I show that these copies generally show very low DNA divergence. Specifically, more than 68% of the transposase genes are identical within a genome. The average number of amino acid replacement substitutions at amino acid replacement sites is Ka = 0.013, that at silent sites is Ks = 0.1. This low intragenomic diversity stands in stark contrast to a much higher divergence of the same insertion sequences among distantly related genomes. Gene conversion among protein-coding genes is unlikely to account for this lack of diversity. The relation between transposition frequencies and silent substitution rates suggests that most insertion sequences in a typical genome are evolutionarily young and have been recently acquired. They may undergo periodic extinction in bacterial lineages. By implication, they are detrimental to their host in the long run. This is also suggested by the highly skewed and patchy distribution of insertion sequences among genomes. In sum, one can think of insertion sequences as slow-acting infectious diseases of cell lineages. PMID- 16373393 TI - Acute myelogenous leukemia in elderly patients not eligible for intensive chemotherapy: the dark side of the moon. AB - BACKGROUND: Acute Myelogenous Leukemia (AML) is a common disease in people aged>60 years. About 50% of the patients are not eligible for aggressive chemotherapy (CT) and are only managed with conservative approaches. Results in this subset of patients have not been reported so far. PATIENTS AND METHODS: We retrospectively evaluated 244 consecutive elderly AML patients (M/F 143/101, median age 72 years, range 60-90) diagnosed at our institution from January 1989 to December 1998 and not eligible for intensive CT. Eighty-nine patients (36.5%) had evolved from previous myelodysplasia (sAML). Fifty-three out of 192 (26.4%) patients with available bone marrow (BM) analysis had oligoblastic leukaemia (blasts<40% and WBC<15x10(9)/l). RESULTS: Sixty-seven patients (27.5%) were managed with supportive treatment only. One hundred seventy-seven patients (72.5%), in order to control disease, received conservative CT, consisting of Hydroxyurea (HU) (127 patients, 71.7%), Cytarabine and 6-Thioguanine (39 patients, 22%) or low-dose cytarabine (11 patients, 6.3%). Median overall survival was 179 days (1-3278) with 50 patients (20.5%) surviving>12 months. Older age (>75 years), poor WHO PS (>2), lower PLT levels (<50x10(9)/l) and higher absolute peripheral blast count (>5x10(9)/l) showed a negative prognostic impact on survival in multivariate analysis. CONCLUSIONS: Our data outline the great heterogeneity of elderly AML patients not eligible for intensive CT. A simple scoring system including easily evaluable parameters, which could distinguish subjects with different prognosis, is proposed. Moreover, randomized studies in order to establish best conservative approaches are warranted. PMID- 16373394 TI - Glutamic acid 709 substitutions highlight the importance of the interaction between androgen receptor helices H3 and H12 for androgen and antiandrogen actions. AB - The mutation of a single amino acid in the ligand binding domain of the human androgen receptor (AR) can induce functional abnormalities; for example, in androgen binding or interactions with coregulators. We report here on the structure/function analysis of the ARE709K substitution that is associated with partial androgen insensitivity syndrome. We introduced several mutations at position 709 and tested the consequences of these changes on AR structure and activity in the presence of androgen and antiandrogens. Our results demonstrate that a strong interaction between helix H12 and residue 709 in H3 is required to obtain a fully functional AR. We show that glutamic acid 709 can be replaced by a bulky tyrosine residue without significant effect on the activation by agonists. In contrast, smaller or linear residues that are unable to maintain a tight interaction with H12 induce a substantial loss of androgen-induced AR activity. We also show that the agonist activity of partial antiandrogens is dependent on the side-chain residue at position 709. Strikingly, the ARE709Y substitution causes the conversion of cyproterone acetate into a pure antiandrogen and bicalutamide into a partial agonist. Together, our structural and functional data reveal the key role of glutamic acid 709 in androgenic and antiandrogenic activities. PMID- 16373395 TI - The corepressors silencing mediator of retinoid and thyroid hormone receptor and nuclear receptor corepressor are involved in agonist- and antagonist-regulated transcription by androgen receptor. AB - We have investigated the role of corepressors SMRT (silencing mediator of retinoid and thyroid hormone receptor) and N-CoR (nuclear receptor corepressor) in transcriptional regulation by androgen receptor (AR) in the LNCaP prostate cancer cell line. Using specific small interference RNAs to knock down SMRT and/or N-CoR in LNCaP cells, we found that SMRT and N-CoR not only mediate antagonist-dependent inhibition of AR activation but also have a widespread role in suppressing agonist-dependent activation of several AR target genes we have tested, including PSA (prostate-specific antigen), TSC22 (TSC22 domain family member 1), NKX3-1 (NK3 transcription factor locus 1), and B2M(beta-2 microglobulin). By sequencing analysis followed by analysis of physical association by chromatin immunoprecipitation assay, we mapped the putative androgen response elements in the NKX3-1 and B2M. Consistent with a role in both antagonist- and agonist-regulated transcription by AR, chromatin immunoprecipitation analysis revealed that both SMRT and N-CoR were recruited by AR to these genes in the presence of either flutamide or R1881. Knocking down SMRT and N-CoR enhanced the recruitment of the coactivators steroid receptor coactivator 1 and p300 by agonist-bound AR and led to increased hyperacetylation of histone H3 and H4, suggesting that the corepressors actively compete with coactivators for binding to agonist-bound AR. Taken together, our data indicate that SMRT and N-CoR corepressors are involved in transcriptional regulation by both agonist- and antagonist-bound AR and regulate the magnitude of hormone response, at least in part, by competing with coactivators. PMID- 16373396 TI - Intronic polymorphisms within TFAP2B regulate transcriptional activity and affect adipocytokine gene expression in differentiated adipocytes. AB - We have identified a gene encoding transcription factor activating enhancer binding protein-2beta (TFAP2B) as a candidate for conferring susceptibility to type 2 diabetes. Although we have also found that TFAP2B was preferentially expressed in adipose cells in a differentiation-dependent manner, the mechanisms by which the gene and gene polymorphisms contribute to conferring susceptibility to the disease have not yet been elucidated. The aim of this study was to evaluate the impact of the polymorphisms within the TFAP2B gene on conferring susceptibility to type 2 diabetes. We identified that a 300-bp DNA fragment in intron 1 of TFAP2B had significant enhancer activity, and the variations of this region affected this enhancer activity in differentiated adipocytes. In an experiment using adenovirus vectors encoding TFAP2B, the expression of TNF-alpha gene was shown to be elevated in the TFAP2B overexpressing cells compared with those in control cells. Furthermore, we demonstrated that the expression of TFAP2B was increased in the adipose tissues of subjects with the disease susceptibility allele, and the plasma levels of TNF-alpha and high sensitivity C reactive peptide were significantly elevated in the patients with the disease susceptibility allele. These results suggest that TFAP2B may contribute to the pathogenesis of type 2 diabetes through regulation of adipocytokine gene expression, and that TFAP2B may be a promising target for treatment or prevention of this disease. PMID- 16373397 TI - A role of the amino-terminal (N) and carboxyl-terminal (C) interaction in binding of androgen receptor to chromatin. AB - The N-terminal domain of AR is known to engage a hormone-dependent interaction with its C-terminal ligand-binding domain, and this N/C interaction is known to modulate AR transcriptional activity. Using Xenopus oocytes as a model system to study transcriptional regulation in chromatin, we found that two previously reported N/C interaction-defective AR mutants, one with deletion of 23FQNLF27(ARDeltaF) and one with a Gly 21 to Glu mutation (ARG21E), were surprisingly inactive in activating transcription from various reporters assembled into chromatin. Further study using chromatin immunoprecipitation assay revealed that these mutants failed to bind both mouse mammary tumor virus-long terminal repeat and prostate-specific antigen enhancer assembled into chromatin. This defect is specific to chromatin because both mutants could bind to a consensus AR response element in vitro and activate transcription driven by mouse mammary tumor virus-long terminal repeat in transient transfection as effective as the wild-type AR. To further substantiate this novel finding, we established 293 cell lines that stably expressed either AR or ARDeltaF mutant in an inducible manner. Using these cell lines, we confirmed by using chromatin immunoprecipitation assay that AR but not ARDeltaF could bind to the endogenous prostate-specific antigen enhancer. Furthermore, we found that the ARDeltaF mutant interacts poorly with Brg1, the ATPase subunit of the chromatin-remodeling factor SWI/SNF. Taken together, our study reveals a novel role of AR N/C interaction in control of AR chromatin binding and suggests a working model that the proper N/C interaction is required for AR to recruit SWI/SNF complex, which in turn remodels chromatin to allow AR to bind to AR response elements in chromatin. PMID- 16373398 TI - Dehydroepiandrosterone mimics acute actions of insulin to stimulate production of both nitric oxide and endothelin 1 via distinct phosphatidylinositol 3-kinase- and mitogen-activated protein kinase-dependent pathways in vascular endothelium. AB - Dehydroepiandrosterone (DHEA) is an adrenal steroid and nutritional supplement that may improve insulin sensitivity. Although steroid hormones classically act by regulating transcription, they may also signal through cell surface receptors to mediate nongenomic actions. Because DHEA may augment insulin sensitivity, we hypothesized that DHEA mimics vascular actions of insulin to acutely activate signaling pathways in endothelium-mediating production of nitric oxide (NO) and endothelin 1 (ET-1). Treatment of bovine aortic endothelial cells with either insulin or DHEA (100 nm, 5 min) stimulated significant increases in NO production (assessed with NO-selective fluorescent dye diaminofluorescein 2). These responses were abolished by pretreatment of cells with L-NAME (nitro-L-arginine methyl ester; NO synthase inhibitor) or wortmannin [phosphatidylinositol (PI) 3 kinase inhibitor]. Under similar conditions, insulin- or DHEA-stimulated phosphorylation of Akt (Ser473) and endothelial nitric oxide synthase (Ser1179) was inhibited by pretreatment of cells with wortmannin (but not MAPK kinase inhibitor PD98059). Acute DHEA treatment also caused phosphorylation of MAPK (Thr202/Tyr204) that was inhibitable by PD98059 (but not wortmannin). DHEA treatment of bovine aortic endothelial cells (100 nM, 5 min) stimulated a 2-fold increase in ET-1 secretion that was abolished by pretreatment of cells with PD98059 (but not wortmannin). We conclude that DHEA has acute, nongenomic actions in endothelium to stimulate production of the vasodilator NO via PI 3-kinase dependent pathways and secretion of the vasoconstrictor ET-1 via MAPK-dependent pathways. Altering the balance between PI 3-kinase- and MAPK-dependent signaling in vascular endothelium may determine whether DHEA has beneficial or harmful effects relevant to the pathophysiology of diabetes. PMID- 16373399 TI - A novel partial agonist of peroxisome proliferator-activated receptor-gamma (PPARgamma) recruits PPARgamma-coactivator-1alpha, prevents triglyceride accumulation, and potentiates insulin signaling in vitro. AB - Partial agonists of peroxisome proliferator-activated receptor-gamma (PPARgamma), also termed selective PPARgamma modulators, are expected to uncouple insulin sensitization from triglyceride (TG) storage in patients with type 2 diabetes mellitus. These agents shall thus avoid adverse effects, such as body weight gain, exerted by full agonists such as thiazolidinediones. In this context, we describe the identification and characterization of the isoquinoline derivative PA-082, a prototype of a novel class of non-thiazolidinedione partial PPARgamma ligands. In a cocrystal with PPARgamma it was bound within the ligand-binding pocket without direct contact to helix 12. The compound displayed partial agonism in biochemical and cell-based transactivation assays and caused preferential recruitment of PPARgamma-coactivator-1alpha (PGC1alpha) to the receptor, a feature shared with other selective PPARgamma modulators. It antagonized rosiglitazone-driven transactivation and TG accumulation during de novo adipogenic differentiation of murine C3H10T1/2 mesenchymal stem cells. The latter effect was mimicked by overexpression of wild-type PGC1alpha but not its LXXLL deficient mutant. Despite failing to promote TG loading, PA-082 induced mRNAs of genes encoding components of insulin signaling and adipogenic differentiation pathways. It potentiated glucose uptake and inhibited the negative cross-talk of TNFalpha on protein kinase B (AKT) phosphorylation in mature adipocytes and HepG2 human hepatoma cells. PGC1alpha is a key regulator of energy expenditure and down regulated in diabetics. We thus propose that selective recruitment of PGC1alpha to favorable PPARgamma-target genes provides a possible molecular mechanism whereby partial PPARgamma agonists dissociate TG accumulation from insulin signaling. PMID- 16373400 TI - RvE1 protects from local inflammation and osteoclast- mediated bone destruction in periodontitis. AB - Periodontitis is a well-appreciated example of leukocyte-mediated bone loss and inflammation that has pathogenic features similar to those observed in other inflammatory diseases such as arthritis. Resolvins are a new family of bioactive products of omega-3 fatty acid transformation circuits initiated by aspirin treatment that counter proinflammatory signals. Because it is now increasingly apparent that local inflammation plays a critical role in many diseases, including cardiovascular disease, atherosclerosis, and asthma, experiments were undertaken to evaluate the actions of the newly described EPA-derived Resolvin E1 (RvE1) in regulation of neutrophil tissue destruction and resolution of inflammation. The actions of an aspirin-triggered lipoxin (LX) analog and RvE1 in a human disease, localized aggressive periodontitis (LAP), were determined. Results indicate that neutrophils from LAP are refractory to anti-inflammatory molecules of the LX series, whereas LAP neutrophils respond to RvE1. In addition, RvE1 specifically binds to human neutrophils at a site that is functionally distinct from the LX receptor. Consistent with these potent actions, topical application of RvE1 in rabbit periodontitis conferred dramatic protection against inflammation induced tissue and bone loss associated with periodontitis. PMID- 16373401 TI - Antigen targeting to MHC class II-enriched late endosomes in colonic epithelial cells: trafficking of luminal antigens studied in vivo in Crohn's colitis patients. AB - In Crohn's disease (CD), colonic epithelial cells (CECs) are suggested to stimulate pro-inflammatory CD4+ T cells. However, the endocytic pathways of luminal antigens involved in underlying MHC class II presentation by CECs remain unknown. Our aim was to elucidate antigen trafficking and associated MHC class II expression in CECs of CD patients in vivo. In CD patients (Crohn's colitis and remission) and healthy controls undergoing colonoscopy, ovalbumin (OVA) was sprayed onto inflamed or healthy mucosa. The subcellular localization of OVA and MHC class II was visualized in biopsies taken from OVA-incubated mucosa using fluorescence and cryoelectron microscopy. Targeting of OVA into late endosomes of CECs was found in healthy (controls and CD in remission) and inflamed mucosa (Crohn's colitis). MHC class II expression in CECs was not detected in healthy mucosa but strongly up-regulated during CD inflammation. Induced MHC class II in CECs was predominantly seen at basolateral membranes and in late endosomes, which were efficiently accessed by internalized OVA. Our data provide in vivo evidence that the endocytic pathway of luminal antigens in CECs of Crohn's colitis patients intersects MHC class II-enriched late endosomes and support the postulated role of CECs in MHC class II-associated antigen presentation during CD. PMID- 16373402 TI - Dimerization and the signal transduction pathway of a small in-frame deletion in the epidermal growth factor receptor. AB - A short, in-frame deletional mutant (E746-A750del) is one of the major mutant forms of epidermal growth factor receptor (EGFR) and has been reported to be a determinant of response to EGFR tyrosine kinase inhibitors such as gefitinib and erlotinib. However, the biological and pharmacological functions of mutational EGFR remain unclear. To clarify these biological functions of deletional EGFR, we examined the cellular response to EGF ligand stimulation. Dimerization and phosphorylation of EGFR were observed without any ligand stimulation in the 293(D) cells transfected with deletional EGFR as compared with those transfected with wild-type EGFR (293(W) cells). When the 293(D) cells were exposed to gefitinib, an immunoblotting analysis revealed remarkable inhibition of AKT phosphorylation but not phospho-p44/42 MAPK. To examine the cellular response in a lung cancer cell line intrinsically expressing deletional EGFR, phospho-EGFR, and downstream reactions were monitored under EGF stimulation with a beads-based mulitiplex assay. EGFR and its downstream proteins were constitutively phosphorylated in the PC-9 cells without any ligand stimulation as compared with A549 lung cancer cells expressing wild-type EGFR. In conclusion, deletional EGFR is constitutively active and phosphorylates p44/42 MAPK and AKT in the cells, although the fact that the EGFR phosphorylation in the PC-9 cells is still modulated by EGF stimulation cannot be ignored. Gefitinib-inhibited phospho-AKT predominantly in deletional EGFR expressing cells. PMID- 16373403 TI - Prostasomes--their effects on human male reproduction and fertility. AB - The prostate is a glandular male accessory sex organ vital for normal fertility. It provides the prostatic component of seminal plasma which nourishes and protects sperm following ejaculation. Prostasomes are small (40-500 nm) membrane bound vesicles produced by epithelial cells lining the prostate acini and are a component of prostatic secretions. Although the existence of these particles has been known for many years, their full function and relevance to reproductive health are largely unknown. Proteomic studies have shown a wide range of proteins (enzymes, structural proteins and novel, unannotated proteins) present in or on the surface of prostasomes providing them with a diverse nature. Interestingly prostasomes are able to fuse with sperm, this event and the associated transfer of proteins lies at the heart of many of their proposed functions. Sperm motility is increased by the presence of prostasomes and their fusion prevents premature acrosome reactions. Prostasomes have been shown to aid protection of sperm within the female reproductive tract because of immunosuppressive, antioxidant and antibacterial properties. Clinically these functions imply a role for prostasomes in male factor infertility. However, the very functions that promote fertility may have negative connotations in later life; recent work has suggested that prostasomes are involved in prostate cancer. Clearly more work is needed to clarify the role of these novel particles and their impact on men's health. PMID- 16373404 TI - A cross-sectional study of the forearm bone density of long-term users of levonorgestrel-releasing intrauterine system. AB - BACKGROUND: There are concerns about the effect of hormonal contraceptives on bone mineral density (BMD), but there is currently no information available on the effect of the levonorgestrel-releasing intrauterine system (LNG-IUS) on BMD. The objective of this study was to compare the BMD of LNG-IUS users with that of controls using the TCu380A intrauterine device (IUD). MATERIALS AND METHODS: A cross-sectional study paired 53 women, aged 25-51 years, who had been using the LNG-IUS for 7 years, with 53 IUD users, according to age (+/-1 year) and body mass index (BMI; kg/m2) (+/-1). BMD was evaluated at the midshaft of the ulna and the distal radius of the nondominant forearm using double X-ray absorptiometry. RESULTS: Mean age of women was 34 years. BMI was slightly over 25 in both groups. Estradiol was normal. Mean BMD was 0.469 +/- 0.008 and 0.467 +/- 0.009 and 0.409 +/- 0.009 and 0.411 +/- 0.009 at the midshaft of the ulna and distal radius in LNG-IUS and IUD users, respectively, without significant differences. CONCLUSIONS: Women aged 25-51 years, using the LNG-IUS for 7 years, had a mean BMD similar to that of the control group of TCu380A IUD users. PMID- 16373405 TI - Cyclic and characteristic expression of phosphorylated Akt in human endometrium and decidual cells in vivo and in vitro. AB - BACKGROUND: Akt is activated by phosphorylation and plays an important role in cell survival and maintenance of structure. METHODS: We investigated whether phosphorylated Akt was characteristically expressed in human endometrium in vivo and whether insulin-like growth factor-I (IGF-I) can activate Akt using cultured decidualized human stromal cells in vitro, using immunohistochemistry and Western blotting analysis. RESULTS: The levels of phosphorylated Akt protein increased markedly in the decidual tissues from ectopic pregnancy. The expression of phosphorylated Akt protein in stromal cells increased with the decidualization. The decidual cells showed strong cytoplasmic staining for phosphorylated Akt. However, cultured decidualized human stromal cells diminished phosphorylated Akt expression compared to control cells. IGF-I administration to decidualized human stromal cells significantly recovered pAkt expression. The effect of IGF-I on decidualized human stromal cells was blocked by an inhibitor of phosphatidylinositol-3 kinase (PI3K) (LY294,002). These results suggest that IGF I may activate Akt via PI3K in human endometrium and decidua. The expression of phosphorylated Akt in stromal cells was only detected in the functional layer, where tissue remodelling occurs during menstruation or implantation. CONCLUSIONS: Akt activation may be involved in cell survival and extracellular matrix remodelling in human endometrium and decidua. PMID- 16373406 TI - Factors affecting endometrial and subendometrial blood flow measured by three dimensional power Doppler ultrasound during IVF treatment. AB - BACKGROUND: No information exists in the literature regarding the factors affecting the blood flow towards the endometrial and subendometrial regions during IVF treatment. METHODS: We examined the effect of women's age, their smoking habits, their type of infertility (i.e. primary or secondary) and parity, causes of infertility and serum estradiol (E2) concentration on endometrial and subendometrial blood flows as measured by a three-dimensional (3D) power Doppler ultrasound during IVF treatment. All patients received a standard long protocol of ovarian stimulation and serum E2 concentration was determined on the day of hCG. 3D ultrasound examination with power Doppler was performed on the day of oocyte collection to determine vascularization index (VI), flow index (FI) and vascularization flow index (VFI) of endometrial and subendometrial regions. RESULTS: The age of women, their smoking habits, their types of infertility and parity and causes of infertility had no effect on the endometrial and subendometrial 3D power Doppler flow indices. There was a negative correlation between serum E(2) concentration and endometrial FI (r = -0.109; P = 0.006). CONCLUSIONS: Endometrial blood flow in IVF treatment was negatively affected by serum E2 concentration only. PMID- 16373407 TI - Clinical use of the perifollicular vascularity assessment in IVF cycles: a pilot study. AB - BACKGROUND: In Italy, a recent law has imposed a ban on the fertilization of more than three oocytes at one time, and all resulting embryos produced must be transferred simultaneously. The aim of the present controlled study was to assess the clinical feasibility and efficacy of the perifollicular vascularity assessment for oocyte selection in IVF cycles. METHODS: Fifty-four young primary infertile non-obese women (27 cases and 27 age- and BMI-matched controls) underwent IVF cycles. The choice of the oocytes to fertilize was performed according to perifollicular vascularization in the experimental group, whereas in the control group, the standard morphologic criteria alone were used. The dose of gonadotrophins used, the dominant follicles obtained, the duration of the ovarian stimulation, the number of oocytes retrieved, the number/quality of oocytes fertilized and of cleaved embryos, cycle cancellation, implantation, clinical pregnancy, ongoing pregnancy, multiple pregnancies and ovarian hyperstimulation syndrome rates were assessed in each group. RESULTS: The assessment of perifollicular vascularity was feasible in 88.9% of cases. No difference between groups was detected in any parameter evaluated. CONCLUSION: Power Doppler assessment of perifollicular vascularity seems to have no clinical utility for oocyte selection in IVF cycles for young infertile women. PMID- 16373408 TI - Warming Echovist contrast medium for hysterocontrastsonography and the effect on the incidence of pelvic pain. A randomized controlled study. AB - BACKGROUND: Hysterocontrastsonography (HyCoSy) is a simple effective investigation of tubal patency. One common side effect is pelvic pain at the time of the procedure. The temperature of the contrast media used may be a causative agent for pelvic pain. This study was designed to assess the effect of warming the contrast to body temperature compared to room temperature on the incidence of pelvic pain. METHODS: A randomized controlled trial was carried out at the Cardiff Assisted Reproduction Unit. Following ethical approval, all women having HyCoSy for tubal assessment were asked if they would be willing to participate in the study. Computer-generated numbers were placed in opaque sealed envelopes, which were opened by the fertility nurse at the time of procedure. The women were randomized into two arms to receive the contrast medium at either room temperature or at body temperature. Randomization was done according to CONSORT guidelines. The pain experienced was then recorded by the patient on a 10-cm visual analogue scale. RESULTS: The mean pain scores in the room temperature and the body temperature arms were 5.1 and 3.86, respectively. This was statistically significant (P = 0.006). CONCLUSION: Warming of Echovist contrast media to body temperature is a simple and effective intervention in reducing discomfort at the time of HyCoSy. PMID- 16373409 TI - Timing luteal phase support in GnRH agonist down-regulated IVF/embryo transfer cycles. AB - BACKGROUND: The aim of this study was to compare the effect of three different times of onset of luteal phase support on ongoing pregnancy rate in infertile patients undergoing treatment with GnRH down-regulated IVF and embryo transfer (IVF/ET). MATERIALS AND METHODS: All consecutive eligible patients planned to undergo their first IVF treatment cycle were randomly allocated to receive vaginal progesterone as luteal support at three different time points, that is, after HCG administration for final oocyte maturation (HCG group), at the day of oocyte retrieval (OR group) or at the day of ET (ET group). The primary endpoint of this study was ongoing pregnancy rate. RESULTS: A total of 385 women were randomized, 130 were allocated to the HCG group, 128 to the OR group and 127 to the ET group. An ongoing pregnancy rate of 20.8% was found in the HCG group versus 22.7 and 23.6% in the OR group and ET group, respectively. The mean number and quality of the retrieved oocytes and the transferred embryos did not differ. CONCLUSION: Based on this data, an 18% difference in ongoing pregnancy rate between the three different times of onset of luteal phase support in GnRH agonist down-regulated IVF/ET cycles can be refuted. Smaller clinically meaningful differences may be present. PMID- 16373410 TI - Do young women with polycystic ovary syndrome show early evidence of preclinical coronary artery disease? AB - BACKGROUND: It is thought that women with polycystic ovary syndrome (PCOS) are at increased risk of developing cardiovascular diseases. METHODS: In this study, we used transthoracic echocardiography to measure coronary flow reserve (CFR) in 28 women with PCOS and in 26 healthy women. RESULTS: The PCOS and the control groups were similar in terms of age (27.1 +/- 4.5 versus 28.8 +/- 4.4 years) and BMI (26.6 +/- 5.7 versus 24.7 +/- 4.4 kg/m2). Fasting insulin levels and homeostasis model assessment insulin resistance index were higher in the PCOS group. LH, the LH/FSH ratio, total testosterone, free testosterone and androstenedione were higher in the PCOS group. FSH, estradiol, prolactin, progesterone, cholesterol, triglyceride and high-sensitive C-reactive protein were similar between the two groups, but homocysteine levels were higher in the PCOS group. Baseline diastolic peak f low velocity (DPFV) (25.0 +/- 4.6 versus 23.3 +/- 2.7 cm/s, P > 0.05), hyperaemic DPFV (71.2 +/- 12.8 versus 73.0 +/- 12.9 cm/s, P > 0.05) and CFR (2.8 +/- 0.8 versus 3.2 +/- 0.8 cm/s, P > 0.05) of the left anterior descending coronary artery were similar between the two groups. CONCLUSION: We conclude that in young women with PCOS and without cardiovascular risk factors, CFR is preserved. PMID- 16373411 TI - Reduced recombination associated with the production of aneuploid sperm in an infertile man: a case report. AB - Studies using gene-linkage analysis have suggested that abnormal recombination during meiosis may lead to the production of aneuploid gametes; however, there is little direct evidence of a link between the two in human males. We analysed spermatocytes in the pachytene stage from a man with extremely high aneuploidy rates in his sperm. Testicular tissue specimens of the infertile man and two vasectomy reversals were processed with immuofluorescent techniques to visualize synaptonemal complex and recombination foci and fluorescent in situ hybridization on spermatocytes and sperm with probes for chromosomes 13, 21, 18, X and Y. We observed no recombination between sex chromosomes in the infertile man, while in two controls, we observed recombination rates of 79.3 and 81.0% between the sex chromosomes. This was associated with a total sex aneuploidy rate of 41.61% in testicular sperm of the infertile man (0.44 and 0.62% in two controls). Recombination on chromosome 21 was reduced in the infertile man, with 10.62% of spermatocytes showing no recombination (0 and 1.67% in two controls), as well as chromosome 13, with 53.98% having < or =1 recombination foci (22.05 and 21.67% in two controls). This was associated with increased aneuploidy for those chromosomes. Chromosome 18 aneuploidy was slightly increased, although there was no apparent decrease in recombination. These results provide the first evidence of both recombination and non-disjunction abnormalities in the same individual. This is also the only reported case of an infertile man who shows no recombination between the sex chromosomes, despite the formation of the sex body. PMID- 16373412 TI - Minireview: timely ovulation: circadian regulation of the female hypothalamo pituitary-gonadal axis. AB - The preovulatory surge in the secretion of LH is timed by a neuroendocrine integrative mechanism that involves ovarian estradiol levels and the endogenous circadian system. Studies in female rats and hamsters have established that the clock in the hypothalamic suprachiasmatic nucleus has a preeminent role in setting the LH surge, and anatomical, physiological, and pharmacological data are revealing the responsible connections between suprachiasmatic nucleus neurons and GnRH and estradiol-receptive areas. Recent investigations show that GnRH and pituitary cells express circadian clock genes that might play a role in the release and reception of the GnRH signal. Analysis of the circadian regulation of the LH surge may provide a model for understanding how multiple neural oscillators function within other neuroendocrine axes. PMID- 16373413 TI - Minireview: recent progress in gonadotropin-releasing hormone neuronal migration. AB - Neurons that synthesize GnRH are critical brain regulators of the reproductive axis, yet they originate outside the brain and must migrate over long distances and varied environments to get to their appropriate positions during development. Many studies, past and present, are providing clues for the types of molecules encountered and movements expected along the migratory route. Recent studies provide real-time views of the behavior of GnRH neurons in the context of in vitro preparations that model those in vivo. Live images provide direct evidence of the changing behavior of GnRH neurons in their different environments, showing that GnRH neurons move with greater frequency and with more alterations in direction after they enter the brain. The heterogeneity of molecular phenotypes for GnRH neurons likely ensures that multiple external factors will be found that regulate the migration of different portions of the GnRH neuronal population at different steps along the route. Molecules distributed in gradients both in the peripheral olfactory system and basal forebrain may be particularly influential in directing the appropriate movement of GnRH neurons along their arduous migration. Molecules that mediate the adhesion of GnRH neurons to changing surfaces may also play critical roles. It is likely that the multiple external factors converge on selective signal transduction pathways to engage the mechanical mechanisms needed to modulate GnRH neuronal movement and ultimately migration. PMID- 16373414 TI - Prostacyclin receptor up-regulates the expression of angiogenic genes in human endometrium via cross talk with epidermal growth factor Receptor and the extracellular signaling receptor kinase 1/2 pathway. AB - Prostacyclin (PGI) is a member of the prostanoid family of lipid mediators that mediates its effects through a seven-transmembrane G protein-coupled receptor (IP receptor). Recent studies have ascertained a role for prostanoid-receptor signaling in angiogenesis. In this study we examined the temporal-spatial expression of the IP receptor within normal human endometrium and additionally explored the signaling pathways mediating the role of IP receptor in activation of target angiogenic genes. Quantitative RT-PCR analysis demonstrated the highest endometrial expression of the IP receptor during the menstrual phase compared with all other stages of the menstrual cycle. Immunohistochemical analysis localized the site of IP receptor expression to the glandular epithelial compartment with stromal and perivascular cell immunoreactivity. Expression of the immunoreactive IP receptor protein was greatest during the proliferative and early secretory phases of the menstrual cycle. To explore the role of the IP receptor in glandular epithelial cells, we used the Ishikawa endometrial epithelial cell line. Stimulation of Ishikawa cells and human endometrial biopsy explants with 100 nm iloprost (a PGI analog) rapidly activated ERK1/2 signaling and induced the expression of proangiogenic genes, basic fibroblast growth factor, angiopoietin-1, and angiopoietin-2, in an epidermal growth factor receptor (EGFR)-dependent manner. Furthermore, EGFR colocalized with IP receptor in the glandular epithelial compartment. These data suggest that PGI-IP interaction within glandular epithelial cells can promote the expression of proangiogenic genes in human endometrium via cross talk with the EGFR. PMID- 16373415 TI - Dendritic processing of excitatory synaptic input in hypothalamic gonadotropin releasing-hormone neurons. AB - The activity of hypothalamic GnRH neurons results in the intermittent release of GnRH required for reproductive function. This intermittent neurosecretory activity has been proposed to reflect integration of intrinsic properties of and synaptic input to GnRH neurons. Determining the relative impact of synaptic inputs at different locations on the GnRH neuron is difficult, if not impossible, using only experimental approaches. Thus, we used electrophysiological recordings and neuronal reconstructions to generate computer models of GnRH neurons to examine the effects of synaptic inputs at varying distances from the soma along dendrites. The parameters of the models were adjusted to duplicate measured passive and active electrophysiology of cells from mouse brain slices. Our morphological findings reinforce the emerging picture of a complex dendritic structure of GnRH neurons. Furthermore, analysis of reduced morphology models indicated that this population of cells is unlikely to exhibit low-frequency tonic spiking in the absence of synaptic input. Finally, applying realistic patterns of synaptic input to modeled GnRH neurons indicates that synapses located more than about 30% of the average dendrite length from the soma cannot drive firing at frequencies consistent with neuropeptide release. Thus, processing of synaptic input to dendrites of GnRH neurons is probably more complex than simple summation. PMID- 16373416 TI - Fetal programming: prenatal testosterone treatment leads to follicular persistence/luteal defects; partial restoration of ovarian function by cyclic progesterone treatment. AB - Prenatal testosterone (T) excess during midgestation leads to estrous cycle defects and polycystic ovaries in sheep. We hypothesized that follicular persistence causes polycystic ovaries and that cyclic progesterone (P) treatment would overcome follicular persistence and restore cyclicity. Twice-weekly blood samples for P measurements were taken from control (C; n = 16) and prenatally T treated (T60; n = 14; 100 mg T, im, twice weekly from d 30-90 of gestation) Suffolk sheep starting before the onset of puberty and continuing through the second breeding season. A subset of C and T60 sheep were treated cyclically with a modified controlled internal drug-releasing device for 13-14 d every 17 d during the first anestrus (CP, 7; TP, 6). Transrectal ovarian ultrasonography was performed for 8 d in the first and 21 d in the second breeding season. Prenatal T excess reduced the number, but increased the duration of progestogenic cycles, reduced the proportion of ewes with normal cycles, increased the proportion of ewes with subluteal cycles, decreased the proportion of ewes with ovulatory cycles, induced the occurrence of persistent follicles, and reduced the number of corpora lutea in those that cycled. Cyclic P treatment in anestrus, which produced one third the P concentration seen during luteal phase of cycle, did not reduce the number of persistent follicles, but increased the number of progestogenic cycles while reducing their duration. These findings suggested that follicular persistence might contribute to the polycystic ovarian morphology. Cyclic P treatment was able to only partially restore follicular dynamics, but this may be related to the low replacement concentrations of P achieved. PMID- 16373417 TI - The transcription factor AP-2beta causes cell enlargement and insulin resistance in 3T3-L1 adipocytes. AB - We have reported the association of variations in the activating protein-2beta (AP-2beta) transcription factor gene with type 2 diabetes. This gene was preferentially expressed in 3T3-L1 adipocytes in a differentiation stage dependent manner, and preliminary experiments showed that subjects with the disease-susceptible allele showed stronger expression in adipose tissue than those without the susceptible allele. Thus, we overexpressed the AP-2beta gene in 3T3-L1 adipocytes to clarify whether AP-2beta might play a crucial role in the pathogenesis of type 2 diabetes through dysregulation of adipocyte function. In cells overexpressing AP-2beta, cells increased in size by accumulation of triglycerides accompanied by enhanced glucose uptake. On the contrary, suppression of AP-2beta expression by small interfering RNA inhibited glucose uptake. Enhancement of glucose uptake by AP-2beta overexpression was attenuated by inhibitors of phospholipase C (PLC) and atypical protein kinase Czeta/lambda (PKCzeta/lambda), but not by a phosphatidylinositol 3-kinase (PI3-K) inhibitor. Consistently, we found activation of PLC and atypical PKC, but not PI3-K, by AP 2beta expression. Furthermore, overexpression of PLCgamma enhanced glucose uptake, and this activation was inhibited by an atypical PKC inhibitor, suggesting that the enhanced glucose uptake may be mediated through PLC and atypical PKCzeta/lambda, but not PI3-K. Moreover, we observed the increased tyrosine phosphorylation of Grb2-associated binder-1 (Gab1) and its association with PLCgamma, indicating that Gab1 may be involved in AP-2beta-induced PLCgamma activation. Finally, AP-2beta overexpression was found to relate to the impaired insulin signaling. We propose that AP-2beta is a candidate gene for producing adipocyte hypertrophy and may relate to the abnormal characteristics of adipocytes observed in obesity. PMID- 16373418 TI - Minireview: kisspeptin neurons as central processors in the regulation of gonadotropin-releasing hormone secretion. AB - The Kiss1 gene encodes a family of peptides called kisspeptins, which bind to the G protein-coupled receptor GPR54. Kisspeptin(s) and its receptor are expressed in the forebrain, and the discovery that mice and humans lacking a functional GPR54 fail to undergo puberty and exhibit hypogonadotropic hypogonadism implies that kisspeptin signaling plays an essential role in reproduction. Studies in several mammalian species have shown that kisspeptins stimulate the secretion of gonadotropins from the pituitary by stimulating the release of GnRH from the forebrain after the activation of GPR54, which is expressed by GnRH neurons. Kisspeptin is expressed abundantly in the arcuate nucleus (Arc) and the anteroventral periventricular nucleus (AVPV) of the forebrain. Both estradiol and testosterone regulate the expression of the Kiss1 gene in the Arc and AVPV; however, the response of the Kiss1 gene to these steroids is exactly opposite between these two nuclei. Estradiol and testosterone down-regulate Kiss1 mRNA in the Arc and up-regulate its expression in the AVPV. Thus, kisspeptin neurons in the Arc may participate in the negative feedback regulation of gonadotropin secretion, whereas kisspeptin neurons in the AVPV may contribute to generating the preovulatory gonadotropin surge in the female. Hypothalamic levels of Kiss1 and GPR54 mRNA increase dramatically at puberty, suggesting that kisspeptin signaling could mediate the neuroendocrine events that trigger the onset of puberty. Together, these observations demonstrate that kisspeptin-GPR54 signaling in the brain serves as an important conduit for controlling GnRH secretion in the developing and adult animal. PMID- 16373420 TI - Minireview: the neuroendocrine regulation of puberty: is the time ripe for a systems biology approach? AB - The initiation of mammalian puberty requires an increase in pulsatile release of GnRH from the hypothalamus. This increase is brought about by coordinated changes in transsynaptic and glial-neuronal communication. As the neuronal and glial excitatory inputs to the GnRH neuronal network increase, the transsynaptic inhibitory tone decreases, leading to the pubertal activation of GnRH secretion. The excitatory neuronal systems most prevalently involved in this process use glutamate and the peptide kisspeptin for neurotransmission/neuromodulation, whereas the most important inhibitory inputs are provided by gamma-aminobutyric acid (GABA)ergic and opiatergic neurons. Glial cells, on the other hand, facilitate GnRH secretion via growth factor-dependent cell-cell signaling. Coordination of this regulatory neuronal-glial network may require a hierarchical arrangement. One level of coordination appears to be provided by a host of unrelated genes encoding proteins required for cell-cell communication. A second, but overlapping, level might be provided by a second tier of genes engaged in specific cell functions required for productive cell-cell interaction. A third and higher level of control involves the transcriptional regulation of these subordinate genes by a handful of upper echelon genes that, operating within the different neuronal and glial subsets required for the initiation of the pubertal process, sustain the functional integration of the network. The existence of functionally connected genes controlling the pubertal process is consistent with the concept that puberty is under genetic control and that the genetic underpinnings of both normal and deranged puberty are polygenic rather than specified by a single gene. The availability of improved high-throughput techniques and computational methods for global analysis of mRNAs and proteins will allow us to not only initiate the systematic identification of the different components of this neuroendocrine network but also to define their functional interactions. PMID- 16373419 TI - Direct effects of aldosterone on cardiomyocytes in the presence of normal and elevated extracellular sodium. AB - It is now recognized that aldosterone is potentially cardiotoxic, although its local effects in the heart are not well understood. We examined the effects of aldosterone on cultured neonatal rat cardiomyocytes in the presence of normal and elevated extracellular Na+ ([Na+]o). We evaluated the intracellular volume of cardiomyocytes in the presence of normal (141 mEq/liter) and elevated (146 mEq/liter) [Na+]o by measuring cell size. Intracellular Na+ was measured using sodium-binding-benzofuran-isophthalate as a fluorescent sodium indicator, and cardiac hypertrophy was assessed using B-type natriuretic peptide transcription and (3)H-leucine incorporation. Cardiomyocytes shrank in the presence of 146 mEq/liter Na+ due to the increased extracellular osmolarity at early phase. Aldosterone (10(-7) mol/liter) mitigated the shrinkage by stimulating Na+ uptake by the cells. This effect of aldosterone was blocked by SM 20220, a Na+/H+ exchanger 1 (NHE1) inhibitor, but not by eplerenone, a mineralocorticoid receptor blocker. Seventy-two hours of exposure to aldosterone in the presence of 146 mEq/liter Na+ led to increases in cardiomyocyte size, 3H-leucine incorporation, and B-type natriuretic peptide and NHE1 transcription that were significantly greater than were seen in the presence of 141 mEq/liter Na+. All but the last were blocked by either eplerenone or SM 20220; the increase in NHE1 transcription was blocked only by eplerenone. Aldosterone exerts a beneficial effect via NHE1 to block cardiomyocyte shrinkage in the presence of elevated [Na+]o at early phase, but long-time exposure to aldosterone in the presence of elevated [Na+]o leads to cardiomyocyte hypertrophy via genomic effects mediated by the mineralocorticoid receptor. PMID- 16373421 TI - Calcitonin gene-related peptide receptor activation by receptor activity modifying protein-1 gene transfer to vascular smooth muscle cells. AB - The neuropeptide calcitonin gene-related peptide (CGRP) is a potent vasodilator that plays a protective role in the cardiovascular system. The receptor for CGRP is an unusual complex of the G protein-coupled calcitonin-like receptor and an obligate receptor activity modifying protein-1 (RAMP1). In this report we provide the first evidence that RAMP1 is rate limiting in vascular smooth muscle cells. Although cultured rat aorta smooth muscle cells express calcitonin like-receptor and RAMP1, we found that CGRP is not a potent activator of the receptor. After overexpression of RAMP1 by adenoviral gene transfer, there was a striking increase in CGRP-induced production of cAMP, with a 75-fold decrease in the EC(50) and a 1.5-fold increase in the maximal response. The biological consequence of this increased receptor activity was observed in three different paradigms. First, RAMP1 gene transfer caused a CGRP-dependent decrease in cell proliferation. Second, RAMP1 and CGRP treatment led to a 3-fold greater free radical-induced reduction in cell number. Finally, RAMP1 gene transfer resulted in a 5-fold CGRP-dependent increase in terminal deoxynucleotidyltransferase mediated deoxyuridine triphosphate nick end labeling-positive apoptotic cells upon serum withdrawal. The mechanisms underlying these effects involved cAMP dependent pathways. We propose that RAMP1 gene transfer may be an effective strategy for increasing the effectiveness of CGRP-induced decrease in restenosis after aortic angioplasty. PMID- 16373422 TI - Redundant and synergistic effect of Cdx-2 and Brn-4 on regulating proglucagon gene expression. AB - Cdx-2 and Brn-4 are recognized as transcriptional activators for the proglucagon gene. These two homeodomain (HD) proteins are able to interact with the correspondent motifs on the G1 enhancer element of proglucagon promoter, separated by only 8 bp. We have examined Brn-4 expression in proglucagon producing cells, isolated hamster Brn-4 cDNA, and localized its activation domain. Ectopic expression of either Cdx-2 or Brn-4 in the pancreatic B cell line In111 provoked it to express proglucagon mRNA, whereas ectopically expressing both of them further stimulated proglucagon mRNA expression in this cell line. Furthermore, Brn-4 was found to synergize with Cdx-2 in activating proglucagon promoter, and the Brn-4 activation domain was not required for this synergistic activation. When the binding site for either Cdx-2 or Brn-4 was mutated, the synergistic activation by these two HD proteins was significantly attenuated, but not abolished. We propose that both cooperative DNA binding and mutual recruitment between Cdx-2 and Brn-4 are involved in this synergistic activation and have detected physical interaction between Cdx-2 and Brn-4 by glutathione-S transferase-fusion protein pull-down assay. Our observations suggest that Cdx-2 and Brn-4, two HD proteins that belong to two different families, exert a synergistic and redundant effect on proglucagon gene expression. PMID- 16373423 TI - Sex-dependent effect of melatonin on systemic erythematosus lupus developed in Mrl/Mpj-Faslpr mice: it ameliorates the disease course in females, whereas it exacerbates it in males. AB - In this study, the effect of chronic administration of melatonin on MRL/MpJ Fas(lpr) mice has been studied. These mice spontaneously develop an autoimmune disease that has many features resembling human systemic lupus erythematosus. In fact, histological studies showed that all female mice and most male mice exhibited glomerular abnormalities, arteritic lesions, and cellular interstitial inflammatory infiltrate ranging from mild to severe patterns. Treatment with melatonin improved the histological pattern in females and worsened it in males. Moreover, female mice treated with melatonin showed a diminution of titers of total serum IgG, IgM, and anti-double-stranded DNA and anti-CII autoantibodies; a decrease in proinflammatory cytokines (IL-2, IL-6, interferon-gamma, TNF-alpha, and IL-1beta), an increase in antiinflammatory cytokines (IL-10), and a decrease in nitrite/nitrate. In male mice, treatment with melatonin exhibited the opposite effect, worsening all the immunological parameters with an elevation of titers of autoantibodies and a prevalence of proinflammatory vs. antiinflammatory cytokines. Similar results were obtained when lymphocytes from spleen and lymph nodes were cultured. Again, melatonin treatment in females decreased proinflammatory cytokines and increased antiinflammatory cytokines produced by lymphocytes; in males, the effect was the opposite. These findings suggest that melatonin action in MRL/MpJ-Fas(lpr) mice is gender dependent, probably through modulation and inhibition of sex hormones. PMID- 16373424 TI - Galectin-1 induces chemokine production and proliferation in pancreatic stellate cells. AB - Galectin-1 is a beta-galactoside-binding lectin. Previous studies have shown that galectin-1 was expressed in fibroblasts of chronic pancreatitis and of desmoplastic reaction associated with pancreatic cancer. These fibroblasts are now recognized as activated pancreatic stellate cells (PSCs). Here, we examined the role of galectin-1 in cell functions of PSCs. PSCs were isolated from rat pancreatic tissue and used in their culture-activated phenotype unless otherwise stated. Expression of galectin-1 was assessed by Western blot analysis, RT-PCR, and immunofluorescent staining. The effects of recombinant galectin-1 on chemokine production and proliferation were evaluated. Activation of transcription factors was assessed by EMSA. Activation of MAPKs was examined by Western blot analysis using anti-phosphospecific antibodies. Galectin-1 was strongly expressed in culture-activated but not freshly isolated PSCs. Recombinant galectin-1 increased proliferation and production of monocyte chemoattractant protein-1 and cytokine-induced neutrophil chemoattractant-1. Galectin-1 activated ERK, JNK, activator protein-1, and NF-kappaB, but not p38 MAPK or Akt. Galectin-1 induced proliferation through ERK and chemokine production mainly through the activation of NF-kappaB and in part by JNK and ERK pathways. These effects of galectin-1 were abolished in the presence of thiodigalactosie, an inhibitor of beta-galactoside binding. In conclusion, our results suggest a role of galectin-1 in chemokine production and proliferation through its beta-galactoside binding activity in activated PSCs. PMID- 16373425 TI - Ileal oxalate absorption and urinary oxalate excretion are enhanced in Slc26a6 null mice. AB - Intestinal oxalate transport, mediated by anion exchange proteins, is important to oxalate homeostasis and consequently to calcium oxalate stone diseases. To assess the contribution of the putative anion transporter (PAT)1 (Slc26a6) to transepithelial oxalate transport, we compared the unidirectional and net fluxes of oxalate across isolated, short-circuited segments of the distal ileum of wild type (WT) mice and Slc26a6 null mice [knockout (KO)]. Additionally, urinary oxalate excretion was measured in both groups. In WT mouse ileum, there was a small net secretion of oxalate (J(net)(Ox) = -5.0 +/-5.0 pmol.cm(-2).h(-1)), whereas in KO mice J(net)(Ox) was significantly absorptive (75 +/- 10 pmol.cm( 2)h.h(-1)), which was the result of a smaller serosal-to-mucosal oxalate flux (J(sm)(Ox)) and a larger mucosal-to-serosal oxalate flux (J(ms)(Ox)). Mucosal DIDS (200 microM) reduced J(sm)(Ox) in WT mice, leading to reversal of the direction of net oxalate transport from secretion to absorption (J(net)(Ox) = 15.0 +/- 5.0 pmol.cm(-2).h(-1)) , but DIDS had no significant effect on KO ileum. In WT mice in the absence of mucosal Cl(-), there were small increases in J(ms)(Ox) and decreases in J(sm)(Ox) that led to a small net oxalate absorption. In KO mice, J(net)(Ox) was 1.5-fold greater in the absence of mucosal Cl(-), due solely to an increase in J(ms)(Ox). Urinary oxalate excretion was about fourfold greater in KO mice compared with WT littermates. We conclude that PAT1 is DIDS sensitive and mediates a significant fraction of oxalate efflux across the apical membrane in exchange for Cl(-); as such, PAT1 represents a major apical membrane pathway mediating J(sm)(Ox). PMID- 16373426 TI - Naturally arising CD4+CD25+ regulatory T cells suppress the expansion of colitogenic CD4+CD44highCD62L- effector memory T cells. AB - Naturally arising CD4+CD25+ regulatory T (T(R)) cells have been shown to prevent and cure murine T cell-mediated colitis. However, their exact mechanism of controlling colitogenic memory CD4+ T cells in in vivo systems excluding the initial process of naive T cell activation and differentiation has not been examined to date. Using the colitogenic effector memory (T(EM)) CD4+ cell mediated colitis model induced by adoptive transfer of colitogenic CD4+CD44(high)CD62L(-) lamina propria (LP) T cells obtained from colitic CD4+CD45RB(high) T cell-transferred mice, we have shown in the present study that CD4+CD25+ T(R) cells are able not only to suppress the development of colitis, Th1 cytokine production, and the expansion of colitogenic LP CD4+ T(EM) cells but also to expand these cells by themselves extensively in vivo. An in vitro coculture assay revealed that CD4+CD25+ T(R) cells proliferated in the presence of IL-2-producing colitogenic LP CD4+ T(EM) cells at the early time point (48 h after culture), followed by the acquisition of suppressive activity at the late time point (96 h after culture). Collectively, these data suggest the distinct timing of the IL-2-dependent expansion of CD4+CD25+ T(R) cells and the their suppressive activity on colitogenic LP CD4+ T(EM) cells. PMID- 16373427 TI - Role of the different isoforms of cyclooxygenase and nitric oxide synthase during gastric ulcer healing in cyclooxygenase-1 and -2 knockout mice. AB - Traditional NSAIDs, selective cyclooxygenase (COX)-2 inhibitors, and inhibitors of nitric oxide synthase (NOS) impair the healing of preexisting gastric ulcers. However, the role of COX-1 (with or without impairment of COX-2) and the interaction between COX and NOS isoforms during healing are less clear. Thus we investigated healing and regulation of COX and NOS isoforms during ulcer healing in COX-1 and COX-2 deficiency and inhibition mouse models. In this study, female wild-type COX-1(-/-) and COX-2(-/-) mice with gastric ulcers induced by cryoprobe were treated intragastrically with vehicle, selective COX-1 (SC-560), COX-2 (celecoxib, rofecoxib, and valdedoxib), and unselective COX (piroxicam) inhibitors. Ulcer healing parameters, mRNA expression, and activity of COX and NOS were quantified. Gene disruption or inhibition of COX-1 did not impair ulcer healing. In contrast, COX-2 gene disruption and COX-2 inhibitors moderately impaired wound healing. More severe healing impairment was found in dual (SC-560 + rofecoxib) and unselective (piroxicam) COX inhibition and combined COX impairment (in COX-1(-/-) mice with COX-2 inhibition and COX-2(-/-) mice with COX 1 inhibition). In the ulcerated repair tissue, COX-2 mRNA in COX-1(-/-) mice, COX 1 mRNA in COX-2(-/-) mice, and, remarkably, NOS-2 and NOS-3 mRNA in COX-impaired mice were more upregulated than in wild-type mice. This study demonstrates that COX-2 is a key mediator in gastric wound healing. In contrast, COX-1 has no significant role in healing when COX-2 is unimpaired but becomes important when COX-2 is impaired. As counterregulatory mechanisms, mRNA of COX and NOS isoforms were increased during healing in COX-impaired mice. PMID- 16373428 TI - Genotypic resistance to lamivudine among hepatitis B virus isolates in Mexico. AB - BACKGROUND: Drug resistance of hepatitis B virus (HBV) is an increasing clinical problem. Resistance to lamivudine in HBV isolates in Mexico has been poorly explored. OBJECTIVES: To characterize the mutation patterns associated with genotypic resistance to lamivudine and their prevalence among HBV isolates in Mexico. MATERIAL AND METHODS: Thirty-nine Mexican HBV isolates were analysed by PCR and line probe assay for detection of genetic variants in the polymerase open reading frame domains B and C (INNO-LiPA HBV DR; INNOGENETICS N. V., Ghent, Belgium). This assay detects wild-type and mutations at codons 180, 204 and 207 of the HBV polymerase gene, and at codon positions 171, 172, 195, 196, 198 and 199 of the HBV surface antigen (HBsAg). HBV isolates were obtained from HBsAg positive serum samples of 15 chronic hepatitis patients, two haemodialysis patients with chronic HBV carriage, 20 men found positive for HBsAg when seeking HIV testing and two AIDS patients with chronic HBV infection. None of the participants had received antiviral therapy. RESULTS: Overall, HBV wild-type was found in 37 (94.9%) out of the 39 isolates studied. Two (5.1%) out of the 39 isolates showed mixed wild-type and mutant populations. These mutations occurred in isolates from one hepatitis patient and one haemodialysis patient. The isolate from the hepatitis patient showed a double mutation at codon positions 180 (L180M) and 204 (M204V), thus a 2.6% prevalence of genotypic resistance to lamivudine was found. The isolate from the haemodialysis patient showed a single mutation at codon position 180 (L180M). The two HBV mutant isolates were further analysed for genotype and both isolates were genotype H. CONCLUSIONS: HBV genotypic resistance to lamivudine exists in Mexican isolates. The results highlight the importance of testing for HBV resistance before treatment and have implications for a more rational use of drugs. PMID- 16373429 TI - Characterization of tetracycline resistance mediated by the efflux pump Tap from Mycobacterium fortuitum. AB - OBJECTIVES: The aim of this study was to characterize the efflux pump Tap from Mycobacterium fortuitum, to test its sensitivity to well known efflux inhibitors, to study the interaction between tetracycline and these compounds and to test the ability of these compounds to overcome efflux pump-mediated tetracycline resistance. For all these studies, we produced Tap protein in Mycobacterium smegmatis. METHODS: Antibiotic susceptibility tests, tetracycline uptake/efflux experiments and checkerboard synergy tests. RESULTS: Tetracycline uptake/efflux experiments showed that Tap protein from M. fortuitum uses the electrochemical gradient across the cytoplasmic membrane to extrude tetracycline from the cell. This efflux activity is inhibited by carbonyl cyanide m-chlorophenylhydrazone (CCCP) and reserpine, consistent with the decrease in MIC observed in antibiotic susceptibility testing in the presence of these inhibitors. Accumulation was not inhibited in experiments in which o-vanadate and chlorpromazine (CPZ) were tested. Inhibitor-treated cells used glycerol as a carbon source to re-establish the electrochemical gradient across the membrane and to restore efflux activity. CCCP, reserpine and CPZ reduced the MIC of tetracycline in the M. smegmatis strain expressing the Tap protein, whereas o-vanadate increased the MIC. We also observed synergy between tetracycline and CPZ or reserpine, and antagonism with o vanadate. CONCLUSIONS: The Tapfor efflux pump uses the electrochemical gradient to extrude tetracycline from the cell. This efflux activity can be inhibited by several compounds. This suggests that similar compounds could be used to overcome antibiotic resistance mediated by efflux pumps. PMID- 16373430 TI - Novel synthetic molecules targeting the bacterial RNA polymerase assembly. AB - OBJECTIVES: Despite extensive functional screening of the bacterial RNA polymerase (RNAP) over the past years, very few novel inhibitors have been reported. We have, therefore, decided to screen with a radically different, non enzymic, protein-protein interaction assay. Our target is the highly conserved RNAP-sigma interaction that is essential for transcription. METHODS: Small molecule inhibitors of the RNAP-sigma interaction were tested for their activity on transcription and on bacteria. RESULTS: These compounds have antibacterial activity against Gram-positive bacteria including multiresistant clinical isolates. CONCLUSIONS: This is, to our knowledge, the first example of a small molecule inhibitor of this interaction. PMID- 16373431 TI - Matrix-dependent structure of GeSi nanocrystals in SiC. AB - It is shown experimentally that GeSi nanocrystals in SiC created after high-dose Ge ion implantation and high-temperature annealing are hexagonal in a hexagonal 4H-SiC matrix and are of cubic structure in a cubic 3C-SiC matrix. This interesting fact could be explained by molecular dynamics as the force of the system nanocrystal-matrix to minimize its interface energy. PMID- 16373432 TI - Future of family medicine workforce in the United States. AB - In response to the Future of Family Medicine Leadership Committee's recommendations(1) The Ohio State University Department of Family Medicine convened 10 faculty development sessions covering the following strategic objectives: (1) Promoting a Sufficient Family Medicine Workforce, (2) the Role of Family Medicine in Academic Health Centers, (3)The New Model of Family Medicine, (4) Electronic Medical Records, (5) Family Medicine Education, (6) Lifelong Learning, (7) Enhancing the Science of Medicine, (8) Quality of Care, (9) Communications, and (10) Leadership and Advocacy. The focus of this editorial is on initiatives and programs to promote a sufficient family medicine workforce. In comparison to other industrialized countries, the United States ranked lowest in primary care functions and lowest in health care outcomes, but highest in health care expenditures. Despite this fact, the trend for United States medical school graduates to select subspecialty careers continues upward. Through collaboration and advocacy, we can all ensure a continued enthusiasm for the selection and retention of family medicine as a career. PMID- 16373433 TI - Are large amounts of sodium stored in an osmotically inactive form during sodium retention? Balance studies in freely moving dogs. AB - Alterations in total body sodium (TBSodium) that covered the range from moderate deficit to large surplus were induced by 10 experimental protocols in 66 dogs to study whether large amounts of Na+ are stored in an osmotically inactive form during Na+ retention. Changes in TBSodium, total body potassium (TBPotassium), and total body water (TBWater) were determined by 4-day balance studies. A rather close correlation was found between individual changes in TBSodium and those in TBWater (r2 = 0.83). Changes in TBSodium were often accompanied by changes in TBPotassium. Taking changes of both TBSodium and TBPotassium into account, the correlation with TBWater changes became very close (r2 = 0.93). The sum of changes in TBSodium and TBPotassium was accompanied by osmotically adequate TBWater changes, and plasma osmolality remained unchanged. Calculations reveal that even moderate TBSodium changes often included substantial Na+/K+ exchanges between extracellular and cellular space. The results support the theory that osmocontrol effectively adjusts TBWater to the body's present content of the major cations, Na+ and K+, and do not support the notion that, during Na+ retention, large portions of Na+ are stored in an osmotically inactive form. Furthermore, the finding that TBSodium changes are often accompanied by TBPotassium changes and also include Na+/K+ redistributions between fluid compartments suggests that cells may serve as readily available Na+ store. This Na+ storage, however, is osmotically active, since osmotical equilibration is achieved by opposite redistribution of K+. PMID- 16373434 TI - Postprandial increase of oleoylethanolamide mobilization in small intestine of the Burmese python (Python molurus). AB - Oleoylethanolamide (OEA) is an endogenous lipid mediator that inhibits feeding in rats and mice by activating the nuclear receptor peroxisome proliferator activated receptor-alpha (PPAR-alpha). In rodents, intestinal OEA levels increase about threefold upon refeeding, a response that may contribute to the induction of between-meal satiety. Here, we examined whether feeding-induced OEA mobilization also occurs in Burmese pythons (Python molurus), a species of ambush hunting snakes that consume huge meals after months of fasting and undergo massive feeding-dependent changes in gastrointestinal hormonal release and gut morphology. Using liquid chromatography/mass spectrometry (LC/MS), we measured OEA levels in the gastrointestinal tract of fasted (28 days) and fed (48 h after feeding) pythons. We observed a nearly 300-fold increase in OEA levels in the small intestine of fed compared with fasted animals (322 +/- 121 vs. 1 +/- 1 pmol/mg protein, n = 3-4). In situ OEA biosynthesis was suggested by the concomitant increase of N-acyl phosphatidylethanolamine species that serve as potential biosynthetic precursors for OEA. Furthermore, we observed a concomitant increase in saturated, mono- and diunsaturated, but not polyunsaturated fatty acid ethanolamides (FAE) in the small intestine of fed pythons. The identification of OEA and other FAEs in the gastrointestinal tract of Python molurus suggests that this class of lipid messengers may be widespread among vertebrate groups and may represent an evolutionarily ancient means of regulating energy intake. PMID- 16373435 TI - Purinergic receptors in human placenta: evidence for functionally active P2X4, P2X7, P2Y2, and P2Y6. AB - Appropriate regulation of ion transport by the human placental syncytiotrophoblast is important for fetal growth throughout pregnancy. In nonplacental tissues, ion transport can be modulated by extracellular nucleotides that raise intracellular calcium ([Ca2+]i) via activation of purinergic receptors. We tested the hypothesis that purinergic receptors are expressed by human placental cytotrophoblast cells and that their activation by extracellular nucleotides modulates ion (K+) efflux and [Ca2+]i. P2X/P2Y receptor agonists 5 bromouridine 5'-triphosphate (5-BrUTP), ADP, ATP, 2',3'-O-(4-benzoyl benzoyl)adenosine 5'-triphosphate (BzATP), and UTP stimulated 86Rb (K+ tracer) efflux from cultured cytotrophoblast cells at early (mononuclear) or later (multinucleate syncytiotrophoblast-like) stages of differentiation, with ATP and UTP particularly potent. 2-Methylthioadenosine 5'-triphosphate (2-MeS-ATP), and UDP elevated 86Rb efflux only from multinucleated cells. All agonists caused a significant peak and plateau increase in [Ca2+]i, although the magnitude of responses was variable. The effect of BzATP, UTP, and UDP in multinucleated cells was unaffected, and that of ATP partially inhibited, by removal of extracellular Ca2+, implicating P2Y receptor activation. mRNA encoding P2X1, P2X2, P2X4, and P2X7 and P2Y1, P2Y2, P2Y4, P2Y6, and P2Y11 were identified in mono- and multinucleated cells, whereas P2X3 and P2X5 mRNA were absent from all samples. Western blot analysis revealed P2X4, P2X7, P2Y2, and P2Y6 protein in cytotrophoblast cells, but P2Y4 was not detected. On the basis of published agonist selectivity, the data indicate the presence of functionally active P2X4, P2X7, P2Y2, and P2Y6 receptors in cytotrophoblast cells. We propose that activation of these receptors, and subsequent elevation of [Ca2+]i, modulates syncytiotrophoblast homeostasis and/or maternofetal ion exchange in response to extracellular nucleotides. PMID- 16373436 TI - Penile erection requires association of soluble guanylyl cyclase with endothelial caveolin-1 in rat corpus cavernosum. AB - Erectile dysfunction is caused by a variety of pathogenic factors, particularly impaired formation and action of nitric oxide (NO). NO released from nerve endings and corpus cavernosum endothelial cells plays a crucial role in initiating and maintaining increased intracavernous pressure, penile vasodilatation, and penile erection. Classically, these effects are dependent on cGMP synthesized during activation of soluble guanylyl cyclase (sGC) by NO in smooth muscle cells. The enzyme NO synthase in endothelial cells has been localized to caveolae, small invaginations of the plasma membrane rich in cholesterol. Membrane cholesterol depletion impairs acetylcholine-induced relaxation in arteries attributed to an alteration in caveolar structure. It has been shown that sGC may be activated in endothelial caveolae contributing to vasodilation. We hypothesized that caveolae are the platform for sGC/cGMP signaling in cavernosum smooth muscle eliciting erection. Methyl-beta cyclodextrin, a pharmacological tool to deplete membrane cholesterol and disassemble caveolae, impaired rat erectile responses in vivo and cavernosum smooth muscle relaxation induced by the NO donor sodium nitroprusside and the sGC activator 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole in vitro. Methyl-beta cyclodextrin had no effect on cavernosum smooth muscle relaxation induced by NO released upon nerve stimulation or by exogenous cGMP. Furthermore, sGC and caveolin-1, the major coat protein of caveolae, were colocalized in rat corpus cavernosum sinusoidal endothelium. Electron microscopy indicated caveolae disruption in corpus cavernosum treated with methyl-beta-cyclodextrin. In summary, our results provide evidence of compartmentalization of sGC in the caveolae of cavernosal endothelial cells contributing to NO signaling mediating smooth muscle relaxation and erection. PMID- 16373437 TI - Abdominal muscle activity during voiding in female rats with normal or irritated bladder. AB - The aims of the present study were to determine in female rats whether abdominal muscle discharges during normal voiding and to describe the effect of bladder irritation on this visceromotor activity. The sensory pathway of this reflex was also determined. Electromyograms (EMGs) indicated that in awake rats, the abdominal muscle was consistently activated during spontaneous voiding and during voiding induced by saline infusion. Similarly, in anesthetized animals, the muscle discharged during urine expulsion. The abdominal EMG activity was not abolished by hypogastric (Hgnx) or sensory pudendal neurectomy (SPdnx). SPdnx dramatically decreased the intercontraction interval and voided volume. Acetic acid infusion reduced the intercontraction interval and increased bladder contraction duration. It also reduced the pressure threshold for evoking the abdominal EMG response and increased the EMG duration and amplitude. Although SPdnx and Hgnx modified some urodynamic parameters, they did not reverse the acetic acid effect on EMG activity. Thus the afferents activating the visceromotor reflex during normal voiding and the increased reflex in response to acetic acid are probably both carried by the pelvic nerve. Abdominal muscle activity induced by bladder distension has been considered to be a pain marker. However, we conclude that in female rats, the abdominal muscle is reflexively activated during physiological urine expulsion. On the other hand, bladder irritation is marked by an exaggeration of this abdominal visceromotor reflex. PMID- 16373439 TI - Social defeat increases food intake, body mass, and adiposity in Syrian hamsters. AB - Overeating and increases in body and fat mass are the most common responses to day-to-day stress in humans, whereas stressed laboratory rats and mice respond oppositely. Group housing of Syrian hamsters increases body mass, adiposity, and food intake, perhaps due to social confrontation-induced stress. In experiment 1 we asked, Does repeated social defeat increase food intake, body mass, and white adipose tissue (WAT) mass in Syrian hamsters? Male hamsters subjected to the resident-intruder social interaction model and defeated intermittently 15 times over 34 days for 7-min sessions significantly increased their food intake, body mass, and most WAT masses compared with nondefeated controls. Defeat significantly increased terminal adrenal norepinephrine, but not epinephrine, content. In experiment 2 we asked, Are 15 intermittent resident-intruder interactions necessary to increase body mass and food intake? Body mass and food intake of subordinate hamsters defeated only once were similar to those of nondefeated controls, but four or eight defeats similarly and significantly increased these responses. In experiment 3 we asked, Do intermittent defeats increase adiposity and food intake more than consecutive defeats? Four intermittent or consecutive defeats similarly and significantly increased food intake and body mass compared with nondefeated controls, but only intermittent defeats significantly increased all WAT masses. Consecutive defeats significantly increased mesenteric and inguinal WAT masses. Plasma leptin, but not insulin, concentrations were similarly and significantly increased compared with nondefeated controls. Collectively, social defeat, a natural stressor, significantly increased food intake, body mass, and adiposity in Syrian hamsters and may prove useful in determining mechanisms underlying human stress-induced obesity. PMID- 16373438 TI - Anatomical and functional characterization of clock gene expression in neuroendocrine dopaminergic neurons. AB - Oscillations of gene expression and physiological activity in suprachiasmatic nucleus (SCN) neurons result from autoregulatory feedback loops of circadian clock gene transcription factors. In the present experiment, we have determined the pattern of PERIOD1 (PER1), PERIOD2 (PER2), and CLOCK expression within neuroendocrine dopaminergic (DAergic) neurons (NDNs) of ovariectomized (OVX) rats. We have also determined the effects of per1, per2, and clock mRNA knockdown in the SCN with antisense deoxyoligonucleotides (AS-ODN) on DA release from NDNs. Diurnal rhythms of PER1 and PER2 expression in tuberoinfundibular DAergic (TIDA) and periventricular hypophyseal DAergic (PHDA) neurons, peaked at circadian time (CT)18 and CT12, respectively. Rhythms of PER1 expression in tuberhypophyseal neuroendocrine DAergic (THDA) neurons were undetectable. Rhythms of PER2 expression were found in all three populations of NDNs, with greater levels of PER2 expression between CT6 and CT12. AS-ODN injections differentially affected DA turnover in the axon terminals of the median eminence (ME), neural lobe (NL) and intermediate lobe (IL) of the pituitary gland, resulting in a significant decrease in DA release in the early subjective night in the ME (TIDA), a significant increase in DA release at the beginning of the day in the IL (PHDA), and no effect in the NL (THDA). AS-ODN-treatment induced a rhythm of DA concentration in the anterior lobe, with greater DA levels in the middle of the day. These data suggest that clock gene expression, particularly PER1 and PER2, within NDNs may act to modulate diurnal rhythms of DA release from NDNs in the OVX rat. PMID- 16373440 TI - Tissue distribution of UT-A and UT-B mRNA and protein in rat. AB - Mammalian urea transporters are facilitated membrane transport proteins belonging to two families, UT-A and UT-B. They are best known for their role of maintaining the renal inner medullary urinary concentrating gradient. Urea transporters have also been identified in tissues not typically associated with urea metabolism. The purpose of this study was to survey the major organs in rat to determine the distribution of UT-A and UT-B mRNA transcripts and protein forms and determine their cellular localization. Five kidney subregions and 17 extrarenal tissues were screened by Northern blot analysis using two UT-A and three UT-B probes and by Western blot analysis using polyclonal COOH-terminal UT-A and UT-B antibodies. Immunohistochemistry was performed on 16 extrarenal tissues using the same antibodies. In kidney, we detected mRNA transcripts and protein bands consistent with previously-identified UT-A and UT-B isoforms, as well as novel forms. We found that UT-A mRNA and protein are widely expressed in extrarenal tissues in various forms that are different from the known isoforms. We determined the cellular localization of UT-A and UT-B in these tissues. We found that both UT-A and UT-B are ubiquitously expressed as numerous tissue-specific mRNA transcripts and protein forms that are localized to cell membranes, cytoplasm, or nuclei. PMID- 16373441 TI - Time-of-day-dependent effects of bright light exposure on human psychophysiology: comparison of daytime and nighttime exposure. AB - Bright light can influence human psychophysiology instantaneously by inducing endocrine (suppression of melatonin, increasing cortisol levels), other physiological changes (enhancement of core body temperature), and psychological changes (reduction of sleepiness, increase of alertness). Its broad range of action is reflected in the wide field of applications, ranging from optimizing a work environment to treating depressed patients. For optimally applying bright light and understanding its mechanism, it is crucial to know whether its effects depend on the time of day. In this paper, we report the effects of bright light given at two different times of day on psychological and physiological parameters. Twenty-four subjects participated in two experiments (n = 12 each). All subjects were nonsmoking, healthy young males (18-30 yr). In both experiments, subjects were exposed to either bright light (5,000 lux) or dim light <10 lux (control condition) either between 12:00 P.M. and 4:00 P.M. (experiment A) or between midnight and 4:00 A.M. (experiment B). Hourly measurements included salivary cortisol concentrations, electrocardiogram, sleepiness (Karolinska Sleepiness Scale), fatigue, and energy ratings (Visual Analog Scale). Core body temperature was measured continuously throughout the experiments. Bright light had a time-dependent effect on heart rate and core body temperature; i.e., bright light exposure at night, but not in daytime, increased heart rate and enhanced core body temperature. It had no significant effect at all on cortisol. The effect of bright light on the psychological variables was time independent, since nighttime and daytime bright light reduced sleepiness and fatigue significantly and similarly. PMID- 16373444 TI - Exercise-induced reversal of insulin resistance in obese elderly is associated with reduced visceral fat. AB - Exercise improves glucose metabolism and delays the onset and/or reverses insulin resistance in the elderly by an unknown mechanism. In the present study, we examined the effects of exercise training on glucose metabolism, abdominal adiposity, and adipocytokines in obese elderly. Sixteen obese men and women (age = 63 +/- 1 yr, body mass index = 33.2 +/- 1.4 kg/m2) participated in a 12-wk supervised exercise program (5 days/wk, 60 min/day, treadmill/cycle ergometry at 85% of heart rate maximum). Visceral fat (VF), subcutaneous fat, and total abdominal fat were measured by computed tomography. Fat mass and fat-free mass were assessed by hydrostatic weighing. An oral glucose tolerance test was used to determine changes in insulin resistance. Exercise training increased maximal oxygen consumption (21.3 +/- 0.8 vs. 24.3 +/- 1.0 ml.kg(-1).min(-1), P < 0.0001), decreased body weight (P < 0.0001) and fat mass (P < 0.001), while fat-free mass was not altered (P > 0.05). VF (176 +/- 20 vs. 136 +/- 17 cm2, P < 0.0001), subcutaneous fat (351 +/- 34 vs. 305 +/- 28 cm2, P < 0.03), and total abdominal fat (525 +/- 40 vs. 443 +/- 34 cm2, P < 0.003) were reduced through training. Circulating leptin was lower (P < 0.003) after training, but total adiponectin and tumor necrosis factor-alpha remained unchanged. Insulin resistance was reversed by exercise (40.1 +/- 7.7 vs. 27.6 +/- 5.6 units, P < 0.01) and correlated with changes in VF (r = 0.66, P < 0.01) and maximal oxygen consumption (r = -0.48, P < 0.05) but not adipocytokines. VF loss after aerobic exercise training improves glucose metabolism and is associated with the reversal of insulin resistance in older obese men and women. PMID- 16373445 TI - Effect of eccentric exercise on position sense at the human forearm in different postures. AB - This is a study of the ability of blindfolded human subjects to match the position of their forearms before and after eccentric exercise. The hypothesis tested was that the sense of effort contributed to forearm position sense. The fall in force after the exercise was predicted to alter the relationship between effort and force and thereby induce position errors. In the arms-in-front posture, subjects had their unsupported reference arm set to one of two angles from the horizontal, 30 or 60 degrees , and they matched its position by voluntary placement of their other arm. Matching errors were compared with a task where the arms were counterweighted, so could be moved in the vertical plane with minimal effort, and where the arms were moved in the horizontal plane. In these latter two tasks, the intention was to test whether removal of an effort sensation from holding the arm against gravity influenced matching performance. It was found that, although absolute errors for counterweighted and horizontal matching were no larger than for unsupported matching, their standard deviations, 6.1 and 6.8 degrees , respectively, were significantly greater than for unsupported matching (4.6 degrees ), indicating more erratic matching. The eccentric exercise led, the next day, to a fall in maximum voluntary muscle torque of >or=15%. This was accompanied by a significant increase in matching errors for the unsupported matching task from 2.7 +/- 0.5 to 0.8 +/- 0.7 degrees but not for counterweighted (1.4 +/- 0.2 to -0.2 degrees +/- 1.1 degrees ) or horizontal matching (-1.3 +/- 0.7 degrees to -1.8 +/- 0.7 degrees ). This, it is postulated, is because the reduced voluntary torque after exercise was accompanied by a greater effort required to support the arms, leading to larger matching errors. However, effort is only able to provide positional information for unsupported matching where gravity plays a role. In gravity-neutral tasks like counterweighted or horizontal matching, a change in the effort-force relationship after exercise leaves matching accuracy unaffected. PMID- 16373446 TI - Aging-sensitive cellular and molecular mechanisms associated with skeletal muscle hypertrophy. AB - Sarcopenia is an age-related loss of muscle mass and strength. The aged can increase various measures of muscle size and strength in response to resistance exercise (RE), but this may not normalize specific tension. In rats, aging reduces the hypertrophy response and impairs regeneration. In this study, we measured cellular and molecular markers, indicative of muscle hypertrophy, that also respond to acute increases in loading. Comparing 6- and 30-mo-old rats, the aims were to 1) determine whether these markers are altered with age and 2) identify age-sensitive responses to acute RE. The muscles of old rats exhibited sarcopenia involving a deficit in contractile proteins and decreased force generation. The RNA-to-protein ratio was higher in the old muscles, suggesting a decrease in translational efficiency. There was evidence of reduced signaling via components downstream from the insulin/insulin-like growth factor (IGF)-I receptors in old muscles. The mRNA levels of myostatin and suppressor of cytokine signaling 2, negative regulators of muscle mass, were lower in old muscles but did not decrease following RE. RE induced increases in the mRNAs for IGF-I, mechano-growth factor, cyclin D1, and suppressor of cytokine signaling 3 were similar in old and young muscles. RE induced phosphorylation of the IGF-I receptor, and Akt increased in young but not old muscles, whereas that of S6K1 was similar for both. The results of this study indicate that a number of components of intracellular signaling pathways are sensitive to age. As a result, key anticatabolic responses appear to be refractory to the stimuli provided by RE. PMID- 16373447 TI - Automated drug delivery system to control systemic arterial pressure, cardiac output, and left heart filling pressure in acute decompensated heart failure. AB - Pharmacological support with inotropes and vasodilators to control decompensated hemodynamics requires strict monitoring of patient condition and frequent adjustments of drug infusion rates, which is difficult and time-consuming, especially in hemodynamically unstable patients. To overcome this difficulty, we have developed a novel automated drug delivery system for simultaneous control of systemic arterial pressure (AP), cardiac output (CO), and left atrial pressure (Pla). Previous systems attempted to directly control AP and CO by estimating their responses to drug infusions. This approach is inapplicable because of the difficulties to estimate simultaneous AP, CO, and Pla responses to the infusion of multiple drugs. The circulatory equilibrium framework developed previously (Uemura K, Sugimachi M, Kawada T, Kamiya A, Jin Y, Kashihara K, and Sunagawa K. Am J Physiol Heart Circ Physiol 286: H2376-H2385, 2004) indicates that AP, CO, and Pla are determined by an equilibrium of the pumping ability of the left heart (SL), stressed blood volume (V), and systemic arterial resistance (R). Our system directly controls SL with dobutamine, V with dextran/furosemide, and R with nitroprusside, thereby controlling the three variables. We evaluated the efficacy of our system in 12 anesthetized dogs with acute decompensated heart failure. Once activated, the system restored SL, V, and R within 30 min, resulting in the restoration of normal AP, CO, and Pla. Steady-state deviations from target values were small for AP [4.4 mmHg (SD 2.6)], CO [5.4 ml x min(-1) x kg(-1) (SD 2.4)] and Pla [0.8 mmHg (SD 0.6)]. In conclusion, by directly controlling the mechanical determinants of circulation, our system has enabled simultaneous control of AP, CO, and Pla with good accuracy and stability. PMID- 16373448 TI - Reduced exercise-associated response of the GH-IGF-I axis and catecholamines in obese children and adolescents. AB - Obesity blunts catecholamine and growth hormone (GH) responses to exercise in adults, but the effect of obesity on these exercise-associated hormonal responses in children is unclear. Therefore, the aim of the present study was to asses the effect of childhood obesity on the counterregulatory hormonal response to acute exercise. Twenty-five obese children (Ob; body mass index > 95%), and 25 age, gender, and maturity-matched normal-weight controls (NW) participated in the study. Exercise consisted of ten 2-min bouts of constant-cycle ergometry above the anaerobic threshold, with 1-min rest intervals between each bout. Pre-, post , and 120-min postexercise blood samples were collected for circulating components of the GH-IGF-I axis and catecholamines. There were no differences in peak exercise heart rate, serum lactate, and peak O2 uptake normalized to lean body mass between the groups. Obesity attenuated the GH response to exercise (8.9 +/- 1.1 vs. 3.4 +/- 0.7 ng/ml in NW and Ob participants, respectively; P < 0.02). No significant differences in the response to exercise were found for other components of the GH-IGF-I axis. Obesity attenuated the catecholamine response to exercise (epinephrine: 52.5 +/- 12.7 vs. 18.7 +/- 3.7 pg/ml, P < 0.02; norepinephrine: 479.5 +/- 109.9 vs. 218.0 +/- 26.0 pg/ml, P < 0.04; dopamine: 17.2 +/- 2.9 vs. 3.5 +/- 1.9 pg/ml, P < 0.006 in NW and Ob, respectively). Insulin levels were significantly higher in the obese children and dropped significantly after exercise in both groups. Despite the elevated insulin levels and the blunted counterregulatory response, none of the participants developed hypoglycemia. Childhood obesity was associated with attenuated GH and catecholamine response to acute exercise. These abnormalities were compensated for, so that exercise was not associated with hypoglycemia, despite increased insulin levels in obese children. PMID- 16373449 TI - Fos- and Jun-related transcription factors are involved in the signal transduction pathway of mechanical loading in condylar chondrocytes. AB - The chondrocytes of the articular condylar cartilage proliferate, hypertrophy and ultimately undergo apoptosis (programmed cell death), being replaced by osteoblasts. Converging results consolidate activator protein-1 (AP-1) transcription factor as the pivotal downstream effector in the early response of stress-sensitive cells to mechanical loading, and the Fra-1, Fra-2, JunB and JunD members of the AP-1 transcription factor family, as mediators in bone remodelling and apoptotic phenomena. The aim of the present study was to examine the involvement of the Fra-1, Fra-2, JunB and JunD proteins in the biochemical response of functionally loaded mandibular condylar cartilage, and the subsequent initiation of cartilage maturation and apoptotic phenomena. Thirty, female, 14 day-old Wistar rats were assigned to two groups: one group was fed a soft diet and the other a hard diet. At day 21 after weaning, experimental animals from both groups were killed at 6, 12 and 48 hours and their condyles harvested. The condylar cartilage of both groups was immunostained using specific antibodies against Fra-1, Fra-2, JunB and JunD. Statistical analysis of the data revealed over-expression of Fra-1, Fra-2, JunB and JunD proteins in all stages of differentiation of chondrocytes derived from the mandibular condylar cartilage of animals fed on a hard diet. Moreover, the involvement of these proteins significantly increased with time in both groups. Since the aforementioned proteins play key roles in remodelling phenomena of bone and cartilage tissue, influencing pivotal cellular functions such as maturation, differentiation and apoptosis, the results of the present study suggest that mandibular condylar chondrocytes sense functional loading changes and respond by induction of proteins associated with biological phenomena that ultimately influence the growth of the condylar cartilage. PMID- 16373450 TI - Relationship between substance P and interleukin-1beta in gingival crevicular fluid during orthodontic tooth movement in adults. AB - Metabolism by peptidases plays an important role in modulating the levels of biologically-active neuropeptides, while that of substance P (SP), a component of gingival crevicular fluid (GCF), may potentiate the inflammatory process in orthodontic tooth movement. The aim of this study was two-fold: (1) to investigate GCF levels of SP and interleukin-1beta (IL-1beta) during human orthodontic tooth movement, and (2) to determine the correlation coefficients between SP and IL-1beta levels in the GCF. The subjects were 3 males, with a mean age of 21.3 +/- 2.8 years old, and 6 females, with a mean age of 23.1 +/- 2.4 years, undergoing orthodontic movement of a single tooth, with the contralateral tooth used as the control. GCF was sampled at the control and treatment (compression) sites before and 1, 4, 8, 24, 72, 120, and 168 hours after initiation of orthodontic treatment. Prevention of plaque-induced inflammation allowed assessment of the dynamics of mechanically stimulated SP and IL-1beta levels in the GCF, which were determined using enzyme-linked immunosorbent assay (ELISA) kits. GCF levels of SP and IL-1beta for the treated teeth were significantly higher (P < 0.001) than for the corresponding control teeth from 8 to 72 hours, and peaked at 24 hours. These results show that the amounts of SP and IL-1beta in GCF increase with orthodontic tooth movement, and indicate that such increases may be involved in inflammation in response to mechanical stress. PMID- 16373452 TI - Dental transposition as a disorder of genetic origin. AB - A sample of 85 dental transpositions in 75 subjects (27 male, 48 female; mean age at diagnosis 12.25 years) involving both maxillary and mandibular arches was analysed using dental panoramic radiographs and clinical records. Transposition affected the maxillary dentition (76 per cent) more frequently than the mandibular dentition (24 per cent). Unilateral transposition accounted for 88 per cent of cases, with the maxilla being involved more commonly than the mandible. Overall, the most common transposition involved the maxillary canine and first premolar (58 per cent). Considering the jaws in isolation, the canine and first premolar were the most commonly affected teeth in the maxilla (83 per cent) whilst in the mandible, the canine and lateral incisor teeth were most commonly transposed (73 per cent). No significant difference in symmetrical distribution of the unilateral transposition sample occurred. There was evidence of associated hypodontia in 41 per cent of the sample; however, if third molars were excluded, this figure decreased to 25 per cent. Peg-shaped maxillary lateral incisors were judged to be present in 27 per cent of subjects, whilst 41 per cent had retained primary teeth; all of these, except one, were primary canines. Overall, the majority of the sample (76 per cent) demonstrated at least one of the dental anomalies under investigation. Multivariate analysis showed associations between unilateral transposition, gender, and the presence of peg-shaped maxillary lateral incisors; whilst bilateral transposition was more closely associated with gender and the presence of retained primary teeth. There was a poor association between both unilateral and bilateral transposition and hypodontia. Together, these results suggest a mutifactorial aetiology to this disorder, with both genetic and environmental factors playing an important role. PMID- 16373451 TI - Early biofilm formation and the effects of antimicrobial agents on orthodontic bonding materials in a parallel plate flow chamber. AB - Decalcification is a commonly recognized complication of orthodontic treatment with fixed appliances. A technology, based on a parallel plate flow chamber, was developed to investigate early biofilm formation of a strain of Streptococcus sanguis on the surface of four orthodontic bonding materials: glass ionomer cement (Ketac Cem), resin-modified glass ionomer cement (Fuji Ortho LC), chemically-cured composite resin (Concise) and light-cured composite resin (Transbond XT). S. sanguis was used as it is one of the primary colonizers of dental hard surfaces. Artificial saliva was supplied as a source of nutrients for the biofilms. The effects of two commercially available mouthrinses (i.e. a fluoride containing rinse and chlorhexidine) were evaluated. Initial colonization of the bacterium was assessed after 6 hours of growth by the percentage surface coverage (PSC) of the biofilm on the disc surfaces. There were statistically significant differences in bacterial accumulation between different bonding materials (P < 0.05), Concise being the least colonized and Transbond XT being the most colonized by S. sanguis biofilms. All materials pre-treated with 0.05 per cent sodium fluoride mouthrinse showed more than 50 per cent reduction in biofilm formation. The 0.2 per cent chlorhexidine gluconate mouthrinse caused significant reduction of biofilm formation on all materials except Ketac Cem. This in vitro study showed that the use of a chemically-cured composite resin (Concise) reduced early S. sanguis biofilm formation. Also, fluoride had a greater effect in reducing the PSC by S. sanguis biofilms than chlorhexidine. Rinsing with 0.05 per cent sodium fluoride prior to placement of orthodontic appliances is effective in reducing early biofilm formation. PMID- 16373453 TI - Root resorption associated with orthodontic force in inbred mice: genetic contributions. AB - Root resorption (RR) is an unwanted sequela of orthodontic treatment. Despite rigorous investigation, no single factor or group of factors that directly causes RR has been identified. The purpose of this study was to examine the effect of the genotype on susceptibility or resistance to develop RR secondary to orthodontic force. Nine-week-old male mice from eight inbred strains were used and randomly distributed into control (C) or treatment (T) groups as follows: A/J (C = 9,T = 9), C57BL/6J (C = 7,T = 8), C3H/HeJ (C = 8,T = 6), BALB/cJ (C = 8,T = 6), 129P3/J (C = 6,T = 8), DBA/2J (C = 8,T = 9), SJL/J (C = 8,T = 10), and AKR/J (C = 9,T = 8). Each of the treated mice received an orthodontic appliance to tip the maxillary left first molar mesially for 9 days. Histological sections of the tooth were used to determine RR and tartrate resistant acid phosphatase (TRAP) activity. The Wilcoxon ranked-sum non-parametric test was used to evaluate differences between the groups. The results showed that the DBA/2J, BALB/cJ, and 129P3/J inbred mouse strains are highly susceptible to RR, whereas A/J, C57BL/6J and SJL/J mice are much more resistant. The variation in the severity of RR associated with orthodontic force among different inbred strains of mice when age, gender, food, housing, and orthodontic force magnitude/duration are controlled support the hypothesis that susceptibility or resistance to RR associated with orthodontic force is a genetically influenced trait. PMID- 16373454 TI - Determination of Bolton tooth-size ratios by digitization, and comparison with the traditional method. AB - The Bolton Index is one of the most useful calculations for precise orthodontic diagnosis as it shows if there is a correct ratio between dental proportions. However, at times, this calculation is not applied because it is a long and time consuming procedure compared with digital methods. A new digital method for measuring tooth sizes and for calculating the Anterior (ABI) and the Overall (OBI) Bolton Index was tested on 100 sets of study dental casts of the permanent dentition in a Spanish sample and compared with the traditional method. The reproducibility of this digital method versus the traditional one was analysed to determine intra- and inter-examiner measurement errors by calculating the coefficients of variation. The results demonstrated that the digital method provided results comparable with those of the traditional technique, since the regression parameters for each index showed that the correlation coefficients of the two methods were very high and similar to each other: r = 0.976 and r = 0.979 for the ABI and OBI, respectively. The results also showed more discrepancies in the ABI than in the OBI using both methods in this sample. PMID- 16373455 TI - Dynamics of frontal, striatal, and hippocampal systems during rule learning. AB - We examined interactions between frontal, striatal, and hippocampal systems during a rule-learning task. Nineteen healthy young adults solved multiple rule learning problems requiring hypothesis testing while functional magnetic resonance images were obtained. Activity in the head of the caudate peaked early after the beginning of each problem and then dropped rapidly. In contrast, activity in prefrontal cortex areas reached peak values later. These results are in accordance with theories suggesting that the striatum identifies the behavioral context necessary for the frontal lobe to select an appropriate strategy. Striatal and hippocampal systems showed antagonistic patterns of activity: Activation in the anterior hippocampus decreased, whereas caudate activity increased. Good learners showed higher activity in the body and tail of the caudate than poor learners, whereas learning success correlated negatively with activity in the hippocampus. Activation in the head of the caudate correlated negatively with hippocampal activation, indicating a potential mechanism for hippocampal activity reduction. PMID- 16373456 TI - Extraclassical receptive field phenomena and short-range connectivity in V1. AB - Extraclassical receptive field phenomena in V1 are commonly attributed to long range lateral connections and/or extrastriate feedback. We address 2 such phenomena: surround suppression and receptive field expansion at low contrast. We present rigorous computational support for the hypothesis that the phenomena largely result from local short-range (< 0.5 mm) cortical connections and lateral geniculate nucleus input. The neural mechanisms of surround suppression in our simulations operate via (A) enhancement of inhibition, (B) reduction of excitation, or (C) action of both simultaneously. Mechanisms (B) and (C) are substantially more prevalent than (A). We observe, on average, a growth in the spatial summation extent of excitatory and inhibitory synaptic inputs for low contrast stimuli. However, we find this is neither sufficient nor necessary to explain receptive field expansion at low contrast, which usually involves additional changes in the relative gain of these inputs. PMID- 16373457 TI - Basal ganglia functional connectivity based on a meta-analysis of 126 positron emission tomography and functional magnetic resonance imaging publications. AB - The striatum receives projections from the entire cerebral cortex. Different, but not mutually exclusive, models of corticostriatal connectivity have been proposed, including connectivity based on proximity, parallel loops, and a model of a tripartite division of the striatum into motor, associative, and limbic areas. All these models were largely based on studies of anatomic connectivity in nonhuman mammals and lesion studies in animals and humans. Functional neuroimaging has the potential to discern patterns of functional connectivity in humans in vivo. We analyzed the functional connectivity between the cortex and the striatum in a meta-analysis of 126 published functional neuroimaging studies. We mapped the peak activations listed in each publication into stereotaxic space and used standard functional imaging statistical methods to determine which cortical areas were most likely to coactivate with different parts of the striatum. The patterns of functional connectivity between the cortex and the different striatal nuclei are broadly consistent with the predictions of the parallel loop model. The rostrocaudal and dorsoventral patterns of corticostriatal functional connectivity are consistent with the tripartite division of the striatum into motor, associative, and limbic zones. PMID- 16373458 TI - Persistent problems plague 340B program. PMID- 16373459 TI - Genotyping systems for drug-metabolizing enzymes go clinical. PMID- 16373460 TI - Some Medicare Part D medication management programs will use pharmacists. PMID- 16373462 TI - Pharmacology and therapeutic uses of theanine. PMID- 16373463 TI - Complementary and alternative medicine for upper-respiratory-tract infection in children. AB - PURPOSE: Evidence on the efficacy and safety of complementary and alternative medicine (CAM) for the prevention and treatment of upper-respiratory-tract infection (URTI) in children is reviewed. SUMMARY: A search of the literature to June 2005 identified six clinical trials examining the use of herbal medicines and nine trials of other CAM therapies. All articles were critically evaluated for adherence to standards of efficacy and safety research. Echinacea did not reduce the duration and severity of URTI. Andrographis paniculata or echinacea decreased nasal secretions (p < 0.01) but not URTI symptoms. A combination of echinacea, propolis, and ascorbic acid decreased the number of URTI episodes, the duration of symptoms, and the number of days of illness (p < 0.001). Echinacea was associated with a higher frequency of rash compared with placebo (p = 0.008). Neither ascorbic acid nor homeopathy was effective. The efficacy of zinc was not clear, and zinc may be associated with adverse effects in children. Osteopathic manipulation decreased episodes of acute otitis media (p = 0.04) and the need for tympanostomy tube insertion (p = 0.03) in children with recurrent acute otitis media. Stress-management therapy reduced the duration of URTI compared with relaxation therapy with guided imagery or standard care (p < 0.05). CONCLUSION: Current data are generally inadequate to support CAM for the prevention or treatment of URTI in children. PMID- 16373464 TI - Eszopiclone. AB - PURPOSE: The pharmacology, pharmacokinetics, indications, clinical efficacy, adverse effects, drug interactions, dosing, and administration of eszopiclone are discussed. SUMMARY: The pharmacology of eszopiclone is not well understood. Eszopiclone is the S-isomer of racemic zopiclone. The relative bioavailability of oral racemic zopiclone is about 80%. Eszopiclone is rapidly absorbed after oral administration, with peak serum concentrations ranging from 1 to 1.3 hours. The efficacy of eszopiclone has been evaluated in healthy adults, including elderly patients, for the treatment of transient and chronic insomnia. Compared with placebo, eszopiclone has been shown to considerably reduce sleep induction and improve sleep maintenance, duration, quality, and depth, as well as next-day functioning. The most common adverse effects reported are unpleasant taste, headache, and dry mouth. Dosing should be individualized, and the lowest effective dose should be used to minimize the risk of adverse events. The recommended starting dosage for nonelderly patients is 2 mg immediately before bedtime, with adjustment to 3 mg if clinically indicated. Dosage adjustment is necessary in patients with severe hepatic disease and in those receiving concomitant potent cytochrome P-450 isoenzyme 3A4 inhibitors. No dosage adjustment is required for patients with renal dysfunction. The cost of eszopiclone is 3.70 dollars per tablet for all dosage strengths (1-, 2-, and 3-mg tablets). CONCLUSION: Its favorable adverse-effect profile and approved labeling for the treatment of chronic insomnia makes eszopiclone a viable alternative for insomnia treatment. Published data are limited, however, and more clinical trials, including comparator studies, are needed to further evaluate the use of this drug. PMID- 16373465 TI - Aldosterone antagonists in the treatment of heart failure. AB - PURPOSE: The clinical benefits, adverse effects, pharmacokinetics, and recommendations for the appropriate use of the aldosterone antagonists spironolactone and eplerenone in patients with heart failure are reviewed. SUMMARY: Heart failure is a clinical syndrome characterized by the functional inability of the ventricle to meet the metabolic demands of the body. Renal hypoperfusion occurs as a result of reduced cardiac output, resulting in the activation of the renin-angiotensin-aldosterone system, which compensates for the hypoperfusion. However, this contributes to the pathology of the disease by, among other actions, increasing the release of aldosterone. Aldosterone has been shown to cause coronary inflammation, cardiac hypertrophy, myocardial fibrosis, ventricular arrhythmias, and ischemic and necrotic lesions. There are currently two aldosterone antagonists commercially available in the United States, spironolactone and eplerenone. Spironolactone is a nonselective aldosterone antagonist, and eplerenone is selective to the aldosterone receptor. Although numerous clinical trials have evaluated the efficacy of each drug, no studies have directly compared spironolactone and eplerenone. Both have been shown to improve morbidity and mortality in patients with advanced heart failure. Adverse effects of both spironolactone and eplerenone include potentially life threatening hyperkalemia, which can be induced by renal insufficiency, diabetes mellitus, advanced heart failure, advanced age, and concurrent drug therapy. CONCLUSION: Spironolactone and eplerenone are life-saving agents in patients with advanced heart failure and may benefit patients with mild heart failure. Potassium and renal function must be routinely assessed to minimize the risk of life-threatening hyperkalemia. PMID- 16373466 TI - Improving the safety of medication administration using an interactive CD-ROM program. AB - PURPOSE: The impact of an interactive CD-ROM program on the rate of medication administration errors made by nurses was studied. METHODS: This randomized, controlled, nonblinded study was conducted at three community hospitals. Study participants included 30 registered nurses who had at least one year of nursing experience in acute care and who worked on medical or medical-surgical units. Nurses were randomized to an intervention group that completed an interactive CD ROM program on safe medication practices or to a control group. Direct observation was used to determine the baseline (preintervention) and postintervention error rates for both study and control groups. Three categories of errors were defined: deviation from safe administration practices (core 1), preparation and administration errors (core 2), and deviations from prescribed therapy (core 3). An error rate was calculated for each nurse, and the error rates for the study and control groups were based on the average error rate for the nurses in each group. RESULTS: The majority of errors made were core 1 errors. The nurse-level data showed a significant decrease in core 1 error rates between baseline and postintervention periods. Core 2 error rates were higher in the postintervention period, but the increase was not significant. Very few core 3 errors were made by either group during either period. CONCLUSION: An interactive CD-ROM enabled nurses to apply the information learned to identify errors in medication administration and improved adherence to safe medication administration practices. PMID- 16373467 TI - National survey of dietary supplement policies in acute care facilities. AB - PURPOSE: Policies on the use of dietary supplements in acute care facilities were studied. METHODS: A national online survey was conducted in 2004 to determine institutional policies and practices related to the use of dietary supplements. Directors of pharmacy in acute care settings were asked about the role of the pharmacy and therapeutics committee, the use of a nonformulary request process, the use of informed consent, requirements for written orders, mechanisms to identify dietary supplements, recording of supplement use in the pharmacy database, recording of supplement use in the medication administration record, prohibitions on dietary supplement use, storage of dietary supplements, reporting of adverse events, and the use of published references. RESULTS: Usable responses were received from 302 (25.4%) of 1189 pharmacy directors. Policies on dietary supplements were developed and implemented in 62% of facilities, with 38% of respondents indicating that no such policy existed. Policies most commonly required a written order by an authorized prescriber, documentation of use in the medication administration record, and a mechanism for identification of dietary supplements by a health care practitioner prior to use. Thirty percent of respondents with policies reported that they prohibited dietary supplement use, and 9% of those without policies prohibited use in practice. Most institutions allowed the use of a patient's own supply of supplements if ordered by an authorized prescriber. Supplements were most commonly stored at the nurses' station or in a patient drawer. The most commonly used reference was the Internet. Pharmacists had concerns about the consistency of dietary supplement formulations, the lack of FDA review of supplements, and the difficulty of identifying supplements and distinguishing reputable manufacturers. CONCLUSION: A survey of pharmacy directors in acute care facilities revealed that many had no formal policy regarding dietary supplements and had not implemented planning for such a policy. PMID- 16373468 TI - Suitability of written supplemental materials available on the Internet for nonprescription medications. AB - PURPOSE: The suitability, readability, and cultural appropriateness of written supplemental materials available on the Internet for nonprescription medications were examined. METHODS: We videotaped 48 hours of television programming, recording a total of 152 advertisements highlighting 37 unique nonprescription medications. The supplemental materials corresponding to each advertised medication were downloaded and printed in their entirety from each product specific Web site. These materials were assessed using the Suitability Assessment of Materials (SAM) instrument. Total SAM scores were grouped as follows: not suitable (0-39%), adequate (40-69%), and superior (70-100%). The Fry readability formula was used to determine the reading grade level for the materials assessed with the SAM instrument. RESULTS: The mean +/- S.D. SAM score of all materials was 54.9% +/- 0.1% (range, 38-76%). Materials for the majority of drugs (86.5%, n = 32) were rated adequate. Materials for four drugs (10.8%) were rated superior, and the material for one drug was not suitable. While the total SAM scores were adequate for most of the materials evaluated, the majority of materials scored particularly poorly for their reading level (the materials for 81.1% of drugs were not suitable). The materials for 40.9% of drugs used uncommon words. CONCLUSION: Evaluation of the suitability, readability, and cultural appropriateness of written supplemental materials for nonprescription medications available on manufacturer-sponsored Web sites and intended for consumers or patients revealed that SAM scores were adequate for most of the materials; however, many scored poorly in the areas of reading level and used uncommon words. PMID- 16373469 TI - Physicochemical stability of highly concentrated total nutrient admixtures for fluid-restricted patients. AB - PURPOSE: The physicochemical stability of highly concentrated total nutrient admixtures (TNAs) for fluid-restricted patients was studied. METHODS: Five TNAs made from lipid injectable emulsions (50:50 mixture of medium-chain and long chain triglycerides) designed to meet the full nutritional needs of adults with body weights of 40-80 kg were chosen. Protein was included in the TNAs at 1.5 g/kg for each body weight and was supplied from a concentrated 16% mixture containing the essential and non-essential amino acids. All admixtures were contained in ethylene vinyl acetate bags and were aseptically prepared. Triplicate preparations of each TNA were investigated over 30 hours at room temperature by dynamic light scattering (DLS) and light extinction with single particle optical sensing (LE-SPOS). RESULTS: No significant changes in the physicochemical stability of the TNAs were observed by DLS (mean droplet size) or LE-SPOS (large-diameter tail) from time 0 (immediately after compounding) to 30 hours. All TNAs met the mean-droplet-size criteria outlined by USP for 20% lipid injectable emulsions. CONCLUSION: Concentrated TNA formulations made from lipid injectable emulsions were stable for 30 hours at room temperature. PMID- 16373470 TI - Regulation of homeopathic drug products. AB - PURPOSE: The regulation of homeopathic drug products is discussed. SUMMARY: Homeopathy is a system of medicine based on the observation that high doses of pharmacologically active substances cause symptoms when administered to healthy individuals. These same substances, when prepared in very dilute form, may relieve similar symptoms in conditions resulting from different etiologies. Unlike dietary supplements, homeopathic drugs are subject to the Food, Drug, and Cosmetic Act and regulations issued by FDA. Instead of the new-drug-approval process, premarket approval for homeopathic drugs is by way of monograph approval by the Homeopathic Pharmacopoeia Convention of the United States (HPCUS). Monographs are published in the Homeopathic Pharmacopoeia of the United States (HPUS). The criteria for inclusion in the HPUS require that a homeopathic drug product be determined by HPCUS to be safe and effective and to be prepared according to the specifications of the HPUS general pharmacy section. The long history and established nature of homeopathic drug regulation may provide a model for the regulation of dietary supplements. CONCLUSION: Homeopathic drugs in the United States are subject to well-defined regulatory processes that more closely resemble those that apply to allopathic medications than to dietary supplements. PMID- 16373471 TI - Time dependence of the radiation-induced EPR signal in sucrose. AB - Sucrose and common household sugars (e.g. cane) have been studied as dosemeters for a wide variety of applications. However, previous studies of the post irradiation time dependence of irradiated sugar did not include an electron paramagnetic resonance (EPR) reference material. This work employs synthetic ruby as an EPR reference material to remove significant spectrometer/environmental influences on the measured time-dependent changes in the EPR spectral amplitude of irradiated sucrose. As such, these more accurate measurements should replace the previously published data. PMID- 16373472 TI - Computer based determination of source terms for emergency preparedness. PMID- 16373473 TI - Breaking symmetry in protein dimers: designs and functions. AB - Symmetry, and in particular point group symmetry, is generally the rule for the global arrangement between subunits in homodimeric and other oligomeric proteins. The structures of fragments of tropomyosin and bovine fibrinogen are recently published examples, however, of asymmetric interactions between chemically identical chains. Their departures from strict twofold symmetry are based on simple and generalizable chemical designs, but were not anticipated prior to their structure determinations. The current review aims to improve our understanding of the structural principles and functional consequences of asymmetric interactions in proteins. Here, a survey of >100 diverse homodimers has focused on the structures immediately adjacent to the twofold axis. Five regular frameworks in alpha-helical coiled coils and antiparallel beta-sheets accommodate many of the twofold symmetric axes. On the basis of these frameworks, certain sequence motifs can break symmetry in geometrically defined manners. In antiparallel beta-sheets, these asymmetries include register slips between strands of repeating residues and the adoption of different side-chain rotamers to avoid steric clashes of bulky residues. In parallel coiled coils, an axial stagger between the alpha-helices is produced by clusters of core alanines. Such simple designs lead to a basic understanding of the functions of diverse proteins. These functions include regulation of muscle contraction by tropomyosin, blood clot formation by fibrin, half-of-site reactivity of caspase 9, and adaptive protein recognition in the matrix metalloproteinase MMP9. Moreover, asymmetry between chemically identical subunits, by producing multiple equally stable conformations, leads to unique dynamic and self-assembly properties. PMID- 16373474 TI - A new generation of protein display scaffolds for molecular recognition. AB - Engineered antibodies and their fragments are invaluable tools for a vast range of biotechnological and pharmaceutical applications. However, they are facing increasing competition from a new generation of protein display scaffolds, specifically selected for binding virtually any target. Some of them have already entered clinical trials. Most of these nonimmunoglobulin proteins are involved in natural binding events and have amazingly diverse origins, frameworks, and functions, including even intrinsic enzyme activity. In many respects, they are superior over antibody-derived affinity molecules and offer an ever-extending arsenal of tools for, e.g., affinity purification, protein microarray technology, bioimaging, enzyme inhibition, and potential drug delivery. As excellent supporting frameworks for the presentation of polypeptide libraries, they can be subjected to powerful in vitro or in vivo selection and evolution strategies, enabling the isolation of high-affinity binding reagents. This article reviews the generation of these novel binding reagents, describing validated and advanced alternative scaffolds as well as the most recent nonimmunoglobulin libraries. Characteristics of these protein scaffolds in terms of structural stability, tolerance to multiple substitutions, ease of expression, and subsequent applications as specific targeting molecules are discussed. Furthermore, this review shows the close linkage between these novel protein tools and the constantly developing display, selection, and evolution strategies using phage display, ribosome display, mRNA display, cell surface display, or IVC (in vitro compartmentalization). Here, we predict the important role of these novel binding reagents as a toolkit for biotechnological and biomedical applications. PMID- 16373475 TI - Enthalpic and entropic contributions mediate the role of disulfide bonds on the conformational stability of interleukin-4. AB - The role of disulfide bridges in the structure, stability, and folding pathways of proteins has been the subject of wide interest in the fields of protein design and engineering. However, the relative importance of entropic and enthalpic contributions for the stabilization of proteins provided by disulfides is not always clear. Here, we perform a detailed analysis of the role of disulfides in the conformational stability of human Interleukin-4 (IL4), a four-helix bundle protein. In order to evaluate the contribution of two out of the three disulfides to the structure and stability of IL4, two IL4 mutants, C3T-IL4 and C24T-IL4, were used. NMR and ANS binding experiments were compatible with altered dynamics and an increase of the nonpolar solvent-accessible surface area of the folded state of the mutant proteins. Chemical and thermal unfolding experiments followed by fluorescence and circular dichroism revealed that both mutant proteins have lower conformational stability than the wild-type protein. Transition temperatures of unfolding decreased 14 degrees C for C3T-IL4 and 10 degrees C for C24T-IL4, when compared to WT-IL4, and the conformational stability, at 25 degrees C, decreased 4.9 kcal/mol for C3T-IL4 and 3.2 kcal/mol for C24T-IL4. Interestingly, both the enthalpy and the entropy of unfolding, at the transition temperature, decreased in the mutant proteins. Moreover, a smaller change in heat capacity of unfolding was also observed for the mutants. Thus, disulfide bridges in IL4 play a critical role in maintaining the thermodynamic stability and core packing of the helix bundle. PMID- 16373476 TI - Stereoelectronic effects on polyproline conformation. AB - The polyproline type II (PPII) helix is a prevalent conformation in both folded and unfolded proteins, and is known to play important roles in a wide variety of biological processes. Polyproline itself can also form a type I (PPI) helix, which has a disparate conformation. Here, we use derivatives of polyproline, (Pro)10, (Hyp)10, (Flp)10, and (flp)10, where Hyp is (2S,4R)-4-hydroxyproline, Flp is (2S,4R)-4-fluoroproline, and flp is (2S,4S)-4-fluoroproline, to probe for a stereoelectronic effect on the conformation of polyproline. Circular dichroism spectral analyses show that 4R electron-with-drawing substituents stabilize a PPII helix relative to a PPI helix, even in a solvent that favors the PPI conformation, such as n-propanol. The stereochemistry at C4 ordains the relative stability of PPI and PPII helices, as (flp)10 forms a mixture of PPI and PPII helices in water and a PPI helix in n-propanol. The conformational preferences of (Pro)10 are intermediate between those of (Hyp)10/(Flp)10 and (flp)10. Interestingly, PPI helices of (flp)10 exhibit cold denaturation in n-propanol with a value of T(s) near 70 degrees C. Together, these data show that stereoelectronic effects can have a substantial impact on polyproline conformation and provide a rational means to stabilize a PPI or PPII helix. PMID- 16373477 TI - Frequencies of hydrophobic and hydrophilic runs and alternations in proteins of known structure. AB - Patterns of alternation of hydrophobic and polar residues are a profound aspect of amino acid sequences, but a feature not easily interpreted for soluble proteins. Here we report statistics of hydrophobicity patterns in proteins of known structure in a current protein database as compared with results from earlier, more limited structure sets. Previous studies indicated that long hydrophobic runs, common in membrane proteins, are underrepresented in soluble proteins. Long runs of hydrophobic residues remain significantly underrepresented in soluble proteins, with none longer than 16 residues observed. These long runs most commonly occur as buried alpha helices, with extended hydrophobic strands less common. Avoiding aggregation of partially folded intermediates during intracellular folding remains a viable explanation for the rarity of long hydrophobic runs in soluble proteins. Comparison between database editions reveals robustness of statistics on aqueous proteins despite an approximately twofold increase in nonredundant sequences. The expanded database does now allow us to explain several deviations of hydrophobicity statistics from models of random sequence in terms of requirements of specific secondary structure elements. Comparison to prior membrane-bound protein sequences, however, shows significant qualitative changes, with the average hydrophobicity and frequency of long runs of hydrophobic residues noticeably increasing between the database editions. These results suggest that the aqueous proteins of solved structure may represent an essentially complete sample of the universe of aqueous sequences, while the membrane proteins of known structure are not yet representative of the universe of membrane-associated proteins, even by relatively simple measures of hydrophobic patterns. PMID- 16373478 TI - Protein stabilization by specific binding of guanidinium to a functional arginine binding surface on an SH3 domain. AB - Guanidinium hydrochloride (GuHCl) at low concentrations significantly stabilizes the Fyn SH3 domain. In this work, we have demonstrated that this stabilizing effect is manifested through a dramatic (five- to sixfold) decrease in the unfolding rate of the domain with the folding rate being affected minimally. This behavior contrasts to the effect of NaCl, which stabilizes this domain by accelerating the folding rate. These data imply that the stabilizing effect of GuHCl is not predominantly ionic in nature. Through NMR studies, we have identified a specific binding site for guanidinium, and we have determined a dissociation constant of 90 mM for this interaction. The guanidinium-binding site overlaps with a functionally important arginine-binding pocket on the domain surface, and we have shown that GuHCl is a specific inhibitor of the peptide binding activity of the domain. A different SH3 domain possessing a similar arginine-binding pocket is also thermodynamically stabilized by GuHCl. These data suggest that many proteins that normally interact with arginine-containing ligands may also be able to specifically interact with guanidinium. Thus, some caution should be used when using GuHCl as a denaturant in protein folding studies. Since arginine-mediated interactions are often important in the energetics of protein-protein interactions, our observations could be relevant for the design of small molecule inhibitors of protein-protein interactions. PMID- 16373479 TI - Capture of monomeric refolding intermediate of human muscle creatine kinase. AB - Human muscle creatine kinase (CK) is an enzyme that plays an important physiological role in the energy metabolism of humans. It also serves as a typical model for studying refolding of proteins. A study of the refolding and reactivation process of guanidine chloride-denatured human muscle CK is described in the present article. The results show that the refolding process can be divided into fast and slow folding phases and that an aggregation process competes with the proper refolding process at high enzyme concentration and high temperature. An intermediate in the early stage of refolding was captured by specific protein molecules: the molecular chaperonin GroEL and alpha(s)-casein. This intermediate was found to be a monomer, which resembles the "molten globule" state in the CK folding pathway. To our knowledge, this is the first monomeric intermediate captured during refolding of CK. We propose that aggregation is caused by interaction between such monomeric intermediates. Binding of GroEL with this intermediate prevents formation of aggregates by decreasing the concentration of free monomeric intermediates, whereas binding of alpha(s)-casein with this intermediate induces more aggregation. PMID- 16373480 TI - Evidence of reciprocal tertiary interactions between conserved motifs involved in organizing RNA structure essential for internal initiation of translation. AB - Internal ribosome entry site (IRES) elements consist of highly structured RNA regions that determine internal initiation of translation. We have previously shown that the foot-and-mouth disease virus (FMDV) IRES contains a GNRA tetraloop spanning residues G178UAA181. Here we show that tertiary RNA interactions dependent on the GNRA motif determine the structural organization of the central domain. By using mutational analysis in combination with RNA probing, we have identified distant reciprocal interactions between the GNRA motif and the invariant region G240CACG244, termed motif A. Mutations in motif A caused a decrease in IRES activity as severe as the GUAG substitution in the GNRA motif. Substitutions in either GNRA or motif A sequences induced a common reorganization around the conserved R199AAA202 stem-loop, suggesting that the latter contributes to stabilize the GNRA-motif A interaction. This finding was also consistent with a significant increase in the efficiency of RNA-RNA interactions determined in gel shift assays using as probe the hairpin that contains the GNRA motif compared to a transcript encompassing the entire apical region of the central domain. Thus, we propose that the central domain of the FMDV IRES contains a structural conformation essential for IRES activity stabilized by a tertiary contact involving residues in the GNRA tetraloop and motif A conserved sequences. PMID- 16373481 TI - Recombinant viral RdRps can initiate RNA synthesis from circular templates. AB - The crystal structure of the recombinant hepatitis C virus (HCV) RNA-dependent RNA polymerase (RdRp) revealed extensive interactions between the fingers and the thumb subdomains, resulting in a closed conformation with an established template channel that should specifically accept single-stranded templates. We made circularized RNA templates and found that they were efficiently used by the HCV RdRp to synthesize product RNAs that are significantly longer than the template, suggesting that RdRp could exist in an open conformation prior to template binding. RNA synthesis using circular RNA templates had properties similar to those previously documented for linear RNA, including a need for higher GTP concentration for initiation, usage of GTP analogs, sensitivity to salt, and involvement of active-site residues for product formation. Some products were resistant to challenge with the template competitor heparin, indicating that the elongation complexes remain bound to template and are competent for RNA synthesis. Other products were not elongated in the presence of heparin, indicating that the elongation complex was terminated. Lastly, recombinant RdRps from two other flaviviruses and from the Pseudomonas phage phi6 also could use circular RNA templates for RNA-dependent RNA synthesis, although the phi6 RdRp could only use circular RNAs made from the 3'-terminal sequence of the phi6 genome. PMID- 16373482 TI - Protein tagging at rare codons is caused by tmRNA action at the 3' end of nonstop mRNA generated in response to ribosome stalling. AB - It has been believed that protein tagging caused by consecutive rare codons involves tmRNA action at the internal mRNA site. We demonstrated previously that ribosome stalling either at sense or stop codons caused by certain arrest sequences could induce mRNA cleavage near the arrest site, resulting in nonstop mRNAs that are recognized by tmRNA. These findings prompted us to re-examine the mechanism of tmRNA tagging at a run of rare codons. We report here that either AGG or CGA but not AGA arginine rare-codon clusters inserted into a model crp mRNA encoding cAMP receptor protein (CRP) could cause an efficient protein tagging. We demonstrate that more than three consecutive AGG codons are needed to induce an efficient ribosome stalling therefore tmRNA tagging in our system. The tmRNA tagging was eliminated by overproduction of tRNAs corresponding to rare codons, indicating that a scarcity of the corresponding tRNA caused by the rare codon cluster is an important factor for tmRNA tagging. Mass spectrometry analyses of proteins generated in cells lacking or possessing tmRNA encoding a protease-resistant tag sequence indicated that the truncation and tmRNA tagging occur within the cluster of rare codons. Northern and S1 analyses demonstrated that nonstop mRNAs truncated within the rare-codon clusters are detected in cells lacking tmRNA but not in cells expressing tmRNA. We conclude that a ribosome stalled by the rare codon induces mRNA cleavage, resulting in nonstop mRNAs that are recognized by tmRNA. PMID- 16373483 TI - The antisense strand of small interfering RNAs directs histone methylation and transcriptional gene silencing in human cells. AB - To determine mechanistically how siRNAs mediate transcriptional gene silencing (TGS) in human cells, we have measured histone methylation at targeted promoters, the dependency on active transcription, and whether or not both strands of the siRNA are required for siRNA-mediated TGS. We report here that siRNA treatment increases both H3K9 and H3K27 methylation of the targeted EF1A promoter and that this increase is dependent on nuclear specific delivery of the siRNA. We also find that TGS can be directed by the antisense strand alone, and requires active transcription by RNA polymerase II in human cells as evidenced by sensitivity to alpha-amanatin. The observation of antisense strand-specific siRNA-mediated TGS of EF1A was substantiated by targeting the U3 region of the HIV-1 LTR/promoter. Furthermore, we show that the antisense strand of siRNA EF52 associates with the transiently expressed Flag-tagged DNMT3A, the targeted EF1A promoter, and trimethylated H3K27. The observations reported here implicate a functional link between siRNA-mediated targeting of genomic regions (promoters), RNA Pol II function, histone methylation, and DNMT3A and support a paradigm in which the antisense strands of siRNAs alone can direct sequence-specific transcriptional gene silencing in human cells. PMID- 16373484 TI - TarBase: A comprehensive database of experimentally supported animal microRNA targets. AB - MicroRNAs (miRNAs) are approximately 22-nt RNA segments that are involved in the regulation of protein expression primarily by binding to one or more target sites on an mRNA transcript and inhibiting translation. MicroRNAs are likely to factor into multiple developmental pathways, multiple mechanisms of gene regulation, and underlie an array of inherited disease processes and phenotypic determinants. Several computational programs exist to predict miRNA targets in mammals, fruit flies, worms, and plants. However, to date, there is no systematic collection and description of miRNA targets with experimental support. We describe a database, TarBase, which houses a manually curated collection of experimentally tested miRNA targets, in human/mouse, fruit fly, worm, and zebrafish, distinguishing between those that tested positive and those that tested negative. Each positive target site is described by the miRNA that binds it, the gene in which it occurs, the nature of the experiments that were conducted to test it, the sufficiency of the site to induce translational repression and/or cleavage, and the paper from which all these data were extracted. Additionally, the database is functionally linked to several other useful databases such as Gene Ontology (GO) and UCSC Genome Browser. TarBase reveals significantly more experimentally supported targets than even recent reviews claim, thereby providing a comprehensive data set from which to assess features of miRNA targeting that will be useful for the next generation of target prediction programs. TarBase can be accessed at http://www.diana.pcbi.upenn.edu/tarbase. PMID- 16373485 TI - RAKE and LNA-ISH reveal microRNA expression and localization in archival human brain. AB - microRNAs (miRNAs) are small (approximately 22 nucleotide) regulatory RNAs which play fundamental roles in many biological processes. Recent studies have shown that the expression of many miRNAs is altered in various human tumors and some miRNAs may function as oncogenes or tumor suppressor genes. However, with the exception of glioblastoma multiforme, the expression of miRNAs in brain tumors is unknown. Furthermore, methods to profile miRNAs from formalin-fixed, paraffin embedded (FFPE) archival tissues or to study their cellular and subcellular localization in FFPE tissues have been lacking. Here we report the coordinated miRNA expression analysis from the tissue level to the subcellular level, using the RAKE (RNA-primed, array-based, Klenow Enzyme) miRNA microarray platform in conjunction with Locked Nucleic Acid (LNA)-based in situ hybridization (LNA-ISH) on archival FFPE human brains and oligodendroglial tumors. The ability to profile miRNAs from archival tissues at the tissue level, by RAKE microarrays, and at the cellular level by LNA-ISH, will accelerate studies of miRNAs in human diseases. PMID- 16373486 TI - A general two-step strategy to synthesize lariat RNAs. AB - We describe a general and efficient two-step strategy for lariat RNA synthesis. In the first step, a deoxyribozyme synthesizes 2',5'-branched RNA. In the second step, T4 RNA ligase closes the loop that completes the lariat. The loop-closure reaction can form either a natural or unnatural lariat isomer, depending on which of the two 3'-termini of the branched RNA reacts with the lone 5'-end. We demonstrate two approaches to control formation of either lariat isomer. In conjunction with other routes for lariat RNA synthesis, the two-step strategy described here will facilitate biochemical studies that require lariat RNAs of varying nucleotide sequence. PMID- 16373487 TI - Prp8p dissection reveals domain structure and protein interaction sites. AB - We describe a novel approach to characterize the functional domains of a protein in vivo. This involves the use of a custom-built Tn5-based transposon that causes the expression of a target gene as two contiguous polypeptides. When used as a genetic screen to dissect the budding yeast PRP8 gene, this showed that Prp8 protein could be dissected into three distinct pairs of functional polypeptides. Thus, four functional domains can be defined in the 2413-residue Prp8 protein, with boundaries in the regions of amino acids 394-443, 770, and 2170-2179. The central region of the protein was resistant to dissection by this approach, suggesting that it represents one large functional unit. The dissected constructs allowed investigation of factors that associate strongly with the N- or the C terminal Prp8 protein fragments. Thus, the U5 snRNP protein Snu114p associates with Prp8p in the region 437-770, whereas fragmenting Prp8p at residue 2173 destabilizes its association with Aar2p. PMID- 16373488 TI - 40LoVe interacts with Vg1RBP/Vera and hnRNP I in binding the Vg1-localization element. AB - Localizing mRNAs within the cytoplasm gives cells the ability to spatially restrict protein production, a powerful means to regulate gene expression. Localized mRNA is often visible in microscopically observable particles or granules, and the association of mRNA localization with these structures is an indication that particles or granules may be essential to the localization process. Understanding how such structures form will therefore be important for understanding the function of localization RNPs (L-RNPs). We previously identified a novel component of an L-RNP from the Vg1 mRNA from Xenopus oocytes called 40LoVe. 40LoVe interaction with the Vg1-localization element (Vg1LE) was previously shown to be dependent on the VM1 and E2 sequence motifs within the Vg1LE that cross-link to hnRNP I and Vg1RBP/Vera, respectively. We report interaction of these motif-binding proteins with 40LoVe and identify a 40LoVe Xenopus hnRNP D/AUF1 interaction. We further demonstrate that titration of VM1 and E2 motif binding activity in vivo surprisingly suggests that the motif binding proteins have differing roles during Vg1LE-dependent mRNA localization. PMID- 16373489 TI - Natural selection is not required to explain universal compositional patterns in rRNA secondary structure categories. AB - We have encountered an unexpected property of rRNA secondary structures that may generalize to all RNAs. Analysis of 8892 ribosomal RNA sequences and structures from a wide range of species revealed unexpected universal compositional trends. First, different categories of rRNA secondary structure (stems, loops, bulges, and junctions) have distinct, characteristic base compositions. Second, the observed patterns of variation are similar among sequences from large and small rRNA subunits and all domains of life, despite extensive evolutionary divergence. Surprisingly, these differences do not seem to be related to selection for different compositions in different structural categories, but rather relate to the overall composition of the molecule: Randomized RNAs with no evolutionary history show the same structure-dependent compositional biases as rRNAs. These compositional trends may improve the accuracy of RNA secondary structure prediction, because they allow us to compare predicted structures against known compositional preferences. They also suggest caution in interpreting differences in the rate of change of the GC content in different parts of the molecule as evidence of differential selection. PMID- 16373490 TI - A computational screen for mammalian pseudouridylation guide H/ACA RNAs. AB - The box H/ACA RNA gene family is one of the largest non-protein-coding gene families in eukaryotes and archaea. Recently, we developed snoGPS, a computational screening program for H/ACA snoRNAs, and applied it to Saccharomyces cerevisiae. We report here results of extending our method to screen for H/ACA RNAs in multiple large genomes of related species, and apply it to the human, mouse, and rat genomes. Because of the 250-fold larger search space compared to S. cerevisiae, significant enhancements to our algorithms were required. Complementing extensive cloning experiments performed by others, our findings include the detection and experimental verification of seven new mammalian H/ACA RNAs and the prediction of 23 new H/ACA RNA pseudouridine guide assignments. These assignments include four for H/ACA RNAs previously classified as orphan H/ACA RNAs with no known targets. We also determined systematic syntenic conservation among human and mouse H/ACA RNAs. With this work, 82 of 97 ribosomal RNA pseudouridines and 18 of 32 spliceosomal RNA pseudouridines in mammals have been linked to H/ACA guide RNAs. PMID- 16373491 TI - Contributions of Trf4p- and Trf5p-dependent polyadenylation to the processing and degradative functions of the yeast nuclear exosome. AB - The nuclear exosome is involved in a large number of RNA processing and surveillance pathways. RNase III cleavage intermediates destined to be 3' processed or degraded can be detected when the Rrp6p subunit of the nuclear exosome is absent. Here we show that these processing and degradation intermediates are polyadenylated, and that their polyadenylation is dependent on the activity of Trf4p and Trf5p, two variant poly(A) polymerases. Polyadenylation of cleavage intermediates was inhibited when Trf4p was absent, and reduced to various extents in the absence of Trf5p, suggesting that these two poly(A) polymerases play functionally distinct roles in the polyadenylation of these RNA species. Finally, in the absence of Trf4p, we observed 3'-extended forms of the U4 snRNA that are similar to those observed in the absence of Rrp6p. These results suggest that polyadenylation of RNA processing intermediates plays a functional role in RNA processing pathways and is not limited to RNA surveillance functions. PMID- 16373492 TI - The interaction between C75 of tRNA and the A loop of the ribosome stimulates peptidyl transferase activity. AB - Ribosomal variants carrying mutations in active site nucleotides are severely compromised in their ability to catalyze peptide bond formation (PT) with minimal aminoacyl tRNA substrates such as puromycin. However, catalysis of PT by these same ribosomes with intact aminoacyl tRNA substrates is uncompromised. These data suggest that these active site nucleotides play an important role in the positioning of minimal aminoacyl tRNA substrates but are not essential for catalysis per se when aminoacyl tRNAs are positioned by more remote interactions with the ribosome. Previously reported biochemical studies and atomic resolution X-ray structures identified a direct Watson-Crick interaction between C75 of the A-site substrate and G2553 of the 23S rRNA. Here we show that the addition of this single cytidine residue (the C75 equivalent) to puromycin is sufficient to suppress the deficiencies of active site ribosomal variants, thus restoring "tRNA like" behavior to this minimal substrate. Studies of the binding parameters and the pH-dependence of catalysis with this minimal substrate indicate that the interaction between C75 and the ribosomal A loop is an essential feature for robust catalysis and further suggest that the observed effects of C75 on peptidyl transfer activity reflect previously reported conformational rearrangements in this active site. PMID- 16373493 TI - Structural study of the H/ACA snoRNP components Nop10p and the 3' hairpin of U65 snoRNA. AB - The H/ACA small nucleolar ribonucleoprotein (snoRNP) complexes guide the modification of uridine to pseudouridine at conserved sites in rRNA. The H/ACA snoRNPs each comprise a target-site-specific snoRNA and four core proteins, Nop10p, Nhp2p, Gar1p, and the pseudouridine synthase, Cbf5p, in yeast. The secondary structure of the H/ACA snoRNAs includes two hairpins that each contain a large internal loop (the pseudouridylation pocket), one or both of which are partially complementary to the target RNA(s). We have determined the solution structure of an RNA hairpin derived from the human U65 box H/ACA snoRNA including the pseudouridylation pocket and adjacent stems, providing the first three dimensional structural information on these H/ACA snoRNAs. We have also determined the structure of Nop10p and investigated its interaction with RNA using NMR spectroscopy. Nop10p contains a structurally well-defined N-terminal region composed of a beta-hairpin, and the rest of the protein lacks a globular structure. Chemical shift mapping of the interaction of RNA constructs of U65 box H/ACA 3' hairpin with Nop10p shows that the beta-hairpin binds weakly but specifically to RNA. The unstructured region of Nop10p likely interacts with Cbf5p. PMID- 16373494 TI - Topology of three-way junctions in folded RNAs. AB - The three-way junctions contained in X-ray structures of folded RNAs have been compiled and analyzed. Three-way junctions with two helices approximately coaxially stacked can be divided into three main families depending on the relative lengths of the segments linking the three Watson-Crick helices. Each family has topological characteristics with some conservation in the non-Watson Crick pairs within the linking segments as well as in the types of contacts between the segments and the helices. The most populated family presents tertiary interactions between two helices as well as extensive shallow/minor groove contacts between a linking segment and the third helix. On the basis of the lengths of the linking segments, some guidelines could be deduced for choosing a topology for a three-way junction on the basis of a secondary structure. Examples and prediction bas'ed on those rules are discussed. PMID- 16373495 TI - Virus-like particles of the Ty3 retrotransposon assemble in association with P body components. AB - Retroviruses and retrotransposons assemble intracellular immature core particles around a RNA genome, and nascent particles collect in association with membranes or as intracellular clusters. How and where genomic RNA are identified for retrovirus and retrotransposon assembly, and how translation and assembly processes are coordinated is poorly understood. To understand this process, the subcellular localization of Ty3 RNA and capsid proteins and virus-like particles was investigated. We demonstrate that mRNAs, proteins, and virus-like particles of the yeast Ty3 retrotransposon accumulate in association with cytoplasmic P bodies, which are sites of mRNA translation repression, storage, and degradation. Deletions of genes encoding P-body proteins decreased Ty3 transposition and caused changes in the pattern of Ty3 foci, underscoring the biological significance of the association of Ty3 virus-like protein components and P bodies. These results suggest the hypothesis that P-bodies may serve to segregate translation and assembly functions of the Ty3 genomic RNA to promote assembly of virus-like particles. Because Ty3 has features of a simple retrovirus and P-body functions are conserved between yeast and metazoan organisms, these findings may provide insights into host factors that facilitate retrovirus assembly. PMID- 16373496 TI - p54nrb is a component of the snRNP-free U1A (SF-A) complex that promotes pre-mRNA cleavage during polyadenylation. AB - The U1 snRNP-A (U1A) protein has been known for many years as a component of the U1 snRNP. We have previously described a form of U1A present in human cells in significant amounts that is not associated with the U1 snRNP or U1 RNA but instead is part of a novel complex of non-snRNP proteins that we have termed snRNP-free U1A, or SF-A. Antibodies that specifically recognize this complex inhibit in vitro splicing and polyadenylation of pre-mRNA, suggesting that this complex may play an important functional role in these mRNA-processing activities. This finding was underscored by the determination that one of the components of this complex is the polypyrimidine-tract-binding protein-associated splicing factor, PSF. In order to further our studies on this complex and to determine the rest of the components of the SF-A complex, we prepared several stable HeLa cell lines that overexpress a tandem-affinity-purification-tagged version of U1A (TAP-tagged U1A). Nuclear extract was prepared from one of these cell lines, line 107, and affinity purification was performed along with RNase treatment. We have used mass spectrometry analysis to identify the candidate factors that associate with U1A. We have now identified and characterized PSF, p54(nrb), and p68 as novel components of the SF-A complex. We have explored the function of this complex in RNA processing, specifically cleavage and polyadenylation, by performing immunodepletions followed by reconstitution experiments, and have found that p54(nrb) is critical. PMID- 16373497 TI - Non-snRNP U1A levels decrease during mammalian B-cell differentiation and release the IgM secretory poly(A) site from repression. AB - A regulated shift from the production of membrane to secretory forms of Immunoglobulin M (IgM) mRNA occurs during B cell differentiation due to the activation of an upstream secretory poly(A) site. U1A plays a key role in inhibiting the expression of the secretory poly(A) site by inhibiting both cleavage at the poly(A) site and subsequent poly(A) tail addition. However, how the inhibitory effect of U1A is alleviated in differentiated cells, which express the secretory poly(A) site, is not known. Using B cell lines representing different stages of B cell differentiation, we show that the amount of U1A available to inhibit the secretory poly(A) site is reduced in differentiated cells. Undifferentiated B cells have more total U1A than differentiated cells and a greater proportion of this is not associated with the U1snRNP. We show that this is available to inhibit poly(A) addition at the secretory poly(A) site using cold competitor RNA oligos to de-repress poly(A) addition in nuclear extracts from the respective cell lines. In addition, endogenous non-snRNP associated U1A immunopurified from the different cell lines-inhibits poly(A) polymerase activity proportional to U1A recovered, suggesting that available U1A level alone is responsible for changes in its inhibitory effect at the secretory IgM poly (A) site. PMID- 16373498 TI - PTEN negatively regulates neural stem cell self-renewal by modulating G0-G1 cell cycle entry. AB - Previous studies have demonstrated that a small subpopulation of brain tumor cells share key characteristics with neural stem/progenitor cells in terms of phenotype and behavior. These findings suggest that brain tumors might contain "cancer stem cells" that are critical for tumor growth. However, the molecular pathways governing such stem cell-like behavior remain largely elusive. Our previous study suggests that the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) tumor suppressor gene, one of the most frequently mutated genes in glioblastomas, restricts neural stem/progenitor cell proliferation in vivo. In the present study, we sought to determine the role of PTEN in long-term maintenance of stem cell-like properties, cell cycle entry and progression, and growth factor dependence and gene expression. Our results demonstrate an enhanced self-renewal capacity and G(0)-G(1) cell cycle entry and decreased growth factor dependency of Pten null neural/stem progenitor cells. Therefore, loss of PTEN leads to cell physiological changes, which collectively are sufficient to increase the pool of self-renewing neural stem cells and promote their escape from the homeostatic mechanisms of proliferation control. PMID- 16373500 TI - Mapping of orthologous genes in the context of biological pathways: An application of integer programming. AB - Mapping biological pathways across microbial genomes is a highly important technique in functional studies of biological systems. Existing methods mainly rely on sequence-based orthologous gene mapping, which often leads to suboptimal mapping results because sequence-similarity information alone does not contain sufficient information for accurate identification of orthology relationship. Here we present an algorithm for pathway mapping across microbial genomes. The algorithm takes into account both sequence similarity and genomic structure information such as operons and regulons. One basic premise of our approach is that a microbial pathway could generally be decomposed into a few operons or regulons. We formulated the pathway-mapping problem to map genes across genomes to maximize their sequence similarity under the constraint that the mapped genes be grouped into a few operons, preferably coregulated in the target genome. We have developed an integer-programming algorithm for solving this constrained optimization problem and implemented the algorithm as a computer software program, p-map. We have tested p-map on a number of known homologous pathways. We conclude that using genomic structure information as constraints could greatly improve the pathway-mapping accuracy over methods that use sequence-similarity information alone. PMID- 16373499 TI - Inositols prevent and reverse endothelial dysfunction in diabetic rat and rabbit vasculature metabolically and by scavenging superoxide. AB - Endothelial dysfunction (ED) is an early feature of cardiovascular risk and diabetes. Hyperglycemia and hyperlipidemia are causative factors. Excessive endothelial mitochondrial superoxide (ROS) production with hyperglycemia and hyperlipidemia is a key mechanism. Inositol components of an insulin inositol glycan mediator, d-chiro-inositol (DCI) and 3-O-methyl DCI (pinitol), decrease hyperglycemia and hyperlipidemia. We tested whether these, myoinositol and dibutyryl DCI (db-DCI), would prevent or reverse ED in diabetic rats and rabbits. Oral inositols reduced hyperglycemia and hypertriglyceridemia with different potencies and prevented ED in rat aortic rings and mesenteric beds. Inositols added in vitro to five diabetic tissues reversed ED. Relaxation by Ach, NO, and electrical field stimulation was potentiated by inositols in vitro in rabbit penile corpus cavernosa. Inositols in vitro restored impaired contraction by the eNOS inhibitor l-NAME and increased NO effectiveness. DCI and db-DCI decreased elevated ROS in endothelial cells in high glucose and db-DCI reduced PKC activation, hexosamine pathway activity, and advanced glycation end products to basal levels. Xanthine/xanthine oxidase generated superoxide was reduced by superoxide dismutase or inositols, with db-DCI efficacious in a mechanism requiring chelated Fe(3+). Histochemical examination of rat aortic rings for protein SNO demonstrated a decrease in diabetic rings with restoration by inositols. In summary, inositols prevented and reversed ED in rat and rabbit vessels, reduced elevated ROS in endothelial cells, potentiated nitrergic or vasculo-myogenic relaxations, and preserved NO signaling. These effects are related to their metabolic actions, direct superoxide scavenging, and enhancing and protecting NO signaling. Of the inositols tested, db-DCI was most effective. PMID- 16373501 TI - Carbohydrate microarray for profiling the antibodies interacting with Globo H tumor antigen. AB - Understanding the specificity of cell-surface carbohydrates interaction with antibodies and receptors is important for the development of new therapeutics and high-sensitivity diagnostics. This approach is, however, limited to the availability of natural and truncated sequences of the oligosaccharides and the sensitivity of the assay system. Reported here is the synthesis of the cancer antigen Globo H hexasaccharide, an epitope found on the cell surface of breast, prostate, and ovarian cancers, and its truncated sequences by using the programmable one-pot synthesis strategy. The saccharides were then arrayed covalently on glass slides with different density and used for the fluorencense based binding analysis of two monoclonal antibodies against Globo H and the serum from breast cancer patients, to define the specificity of these antibodies. It was shown that the terminal tetrasaccharide binds the monoclonal antibodies equally well as does the hexasaccharide and the fucose residue is required for effective binding. The serum binds both the defucosylated pentasaccharide and the fucosylated hexasaccharide without a significant difference, perhaps because of the polyclonal nature of the serum or the presence of diverse immune responses to different sugar epitopes at various stages. This method requires very small amounts of materials and is more effective and sensitive than the traditional ELISA method, and thus provides another platform to monitor the immune response to carbohydrate epitopes at different stages during differentiation, metastasis, or treatment. PMID- 16373502 TI - Mouse model for noninvasive imaging of HIF prolyl hydroxylase activity: assessment of an oral agent that stimulates erythropoietin production. AB - Many human diseases are characterized by the development of tissue hypoxia. Inadequate oxygenation can cause cellular dysfunction and death. Tissues use many strategies, including induction of angiogenesis and alterations in metabolism, to survive under hypoxic conditions. The heterodimeric transcription factor hypoxia inducible factor (HIF) is a master regulator of genes that promote adaptation to hypoxia. HIF activity is linked to oxygen availability because members of the EGLN family hydroxylate HIFalpha subunits on specific prolyl residues when oxygen is present, which marks them for ubiquitination and proteasomal degradation. We created a mouse that ubiquitously expresses a bioluminescent reporter consisting of firefly luciferase fused to a region of HIF that is sufficient for oxygen dependent degradation. Our validation studies suggest that this mouse will be useful for monitoring hypoxic tissues and evaluating therapeutic agents that stabilize HIF. One such agent, the HIF prolyl hydroxylase inhibitor FG-4383, was active in the liver and kidney after systemic administration as determined by bioluminescence imaging, transcription profiling, and production of erythropoietin, indicating that the HIF transcriptional program can be manipulated in vivo with orally active organic small molecules. PMID- 16373503 TI - Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8+ T cells in vivo. AB - The lifespan and survival of dendritic cells (DC) in vivo are potentially critical to the expansion of T cell immune responses. We have previously reported that DC loaded with specific antigen are rapidly eliminated by cytotoxic T lymphocytes (CTL) in vivo, but the site, mechanism, and consequences of DC elimination were not defined. In this article we show that DC elimination in vivo occurs in a perforin-dependent manner and does not require IFN-gamma or the presence of CD4(+)CD25(+) regulatory T cells. Most importantly, failure to eliminate DC had profound consequences on the CTL immune response. Perforin deficient mice showed a progressive increase in the numbers of antigen-specific CD8(+) T cells after repeated immunizations with DC. In contrast, in control mice the number of antigen-specific CD8(+) T cells did not notably increase with repeated immunizations. Lastly, we also show that CTL-mediated elimination of DC occurs in peripheral tissues but not in the lymph node. Our data suggest that CTL act as "gatekeepers" that control access of antigen-loaded DC into the lymph node, thereby preventing continued expansion of antigen-specific T cells. PMID- 16373504 TI - Stimulation of muscarinic receptors mimics experience-dependent plasticity in the honey bee brain. AB - Honey bees begin life working in the hive. At approximately 3 weeks of age, they shift to visiting flowers to forage for pollen and nectar. Foraging is a complex task associated with enlargement of the mushroom bodies, a brain region important in insects for certain forms of learning and memory. We report here that foraging bees had a larger volume of mushroom body neuropil than did age-matched bees confined to the hive. This result indicates that direct experience of the world outside the hive causes mushroom body neuropil growth in bees. We also show that oral treatment of caged bees with pilocarpine, a muscarinic agonist, induced an increase in the volume of the neuropil similar to that seen after a week of foraging experience. Effects of pilocarpine were blocked by scopolamine, a muscarinic antagonist. Our results suggest that signaling in cholinergic pathways couples experience to structural brain plasticity. PMID- 16373505 TI - Tyrosine phosphorylation acts as a molecular switch to full-scale activation of the eIF2alpha RNA-dependent protein kinase. AB - Phosphorylation of the alpha-subunit of translation eukaryotic initiation factor 2 (eIF2) leads to the inhibition of protein synthesis in response to diverse conditions of stress. Serine/threonine RNA-dependent protein kinase (PKR) is an eIF2alpha kinase family member induced by type I IFN and activated in response to dsRNA or virus infection. Herein, we demonstrate that human PKR is a dual specificity kinase phosphorylated at Y101, Y162 and Y293 in vitro and in vivo. Site-specific tyrosine phosphorylation is essential for efficient dsRNA-binding, dimerization, kinase activation and eIF2alpha phosphorylation of PKR. Biologically, tyrosine phosphorylation of PKR mediates the antiviral and antiproliferative properties of the kinase through its ability to control translation. Our data demonstrate an important role of tyrosine phosphorylation in biochemical and biological processes caused or mediated by the activation of the eIF2alpha kinase PKR. PMID- 16373506 TI - Multiple sensors control reciprocal expression of Pseudomonas aeruginosa regulatory RNA and virulence genes. AB - The opportunistic pathogen Pseudomonas aeruginosa is responsible for a wide range of acute and chronic infections. The transition to chronic infections is accompanied by physiological changes in the bacteria favoring formation of biofilm communities. Here we report the identification of LadS, a hybrid sensor kinase that controls the reciprocal expression of genes for type III secretion and biofilm-promoting polysaccharides. Domain organization of LadS and the range of LadS-controlled genes suggest that it counteracts the activities of another sensor kinase, RetS. These two pathways converge by controlling the transcription of a small regulatory RNA, RsmZ. This work identifies a previously undescribed signal transduction network in which the activities of signal-receiving sensor kinases LadS, RetS, and GacS regulate expression of virulence genes associated with acute or chronic infection by transcriptional and posttranscriptional mechanisms. PMID- 16373507 TI - Silent plateau potentials, rhythmic bursts, and pacemaker firing: three patterns of activity that coexist in quadristable subthalamic neurons. AB - Subthalamic neurons display uncommon intrinsic behaviors that are likely to contribute to the motor and cognitive functions of the basal ganglia and to many of its disorders. Here, we report silent plateau potentials in these cells. These plateau responses start with a transient burst of action potentials that quickly diminish in amplitude because of spike inactivation and current shunt. The resulting interruption of spiking reveals a stable depolarization (up state) that clamps the cell membrane potential near -40 mV for several seconds. These plateau potentials coexist in single subthalamic neurons with more familiar patterns of burst and pacemaker firing. Within a narrow range of baseline membrane potentials (-67 to -60 mV), depolarization abruptly switches single cells from bistable to rhythmic bursts or tonic firing modes, thus selecting entirely distinct algorithms for integrating cortical and pallidal synaptic inputs. PMID- 16373508 TI - Premature ovarian failure in androgen receptor-deficient mice. AB - Premature ovarian failure (POF) syndrome, an early decline of ovarian function in women, is frequently associated with X chromosome abnormalities ranging from various Xq deletions to complete loss of one of the X chromosomes. However, the genetic locus responsible for the POF remains unknown, and no candidate gene has been identified. Using the Cre/LoxP system, we have disrupted the mouse X chromosome androgen receptor (Ar) gene. Female AR(-/-) mice appeared normal but developed the POF phenotype with aberrant ovarian gene expression. Eight-week-old female AR(-/-) mice are fertile, but they have lower follicle numbers and impaired mammary development, and they produce only half of the normal number of pups per litter. Forty-week-old AR(-/-) mice are infertile because of complete loss of follicles. Genome-wide microarray analysis of mRNA from AR(-/-) ovaries revealed that a number of major regulators of folliculogenesis were under transcriptional control by AR. Our findings suggest that AR function is required for normal female reproduction, particularly folliculogenesis, and that AR is a potential therapeutic target in POF syndrome. PMID- 16373509 TI - Complementary base-pair-facilitated electron tunneling for electrically pinpointing complementary nucleobases. AB - Molecular tips in scanning tunneling microscopy can directly detect intermolecular electron tunneling between sample and tip molecules and reveal the tunneling facilitation through chemical interactions that provide overlap of respective electronic wave functions, that is, hydrogen-bond, metal-coordination bond, and charge-transfer interactions. Nucleobase molecular tips were prepared by chemical modification of underlying metal tips with thiol derivatives of adenine, guanine, cytosine, and uracil and the outmost single nucleobase adsorbate probes intermolecular electron tunneling to or from a sample nucleobase molecule. We found that the electron tunneling between a sample nucleobase and its complementary nucleobase molecular tip was much facilitated compared with its noncomplementary counterpart. The complementary nucleobase tip was thereby capable of electrically pinpointing each nucleobase. Chemically selective imaging using molecular tips may be coined "intermolecular tunneling microscopy" as its principle goes and is of general significance for novel molecular imaging of chemical identities at the membrane and solid surfaces. PMID- 16373511 TI - Multiple stepwise refolding of immunoglobulin domain I27 upon force quench depends on initial conditions. AB - Mechanical folding trajectories for polyproteins starting from initially stretched conformations generated by single-molecule atomic force microscopy experiments [Fernandez, J. M. & Li, H. (2004) Science 303, 1674-1678] show that refolding, monitored by the end-to-end distance, occurs in distinct multiple stages. To clarify the molecular nature of folding starting from stretched conformations, we have probed the folding dynamics, upon force quench, for the single I27 domain from the muscle protein titin by using a C(alpha)-Go model. Upon temperature quench, collapse and folding of I27 are synchronous. In contrast, refolding from stretched initial structures not only increases the folding and collapse time scales but also decouples the two kinetic processes. The increase in the folding times is associated primarily with the stretched state to compact random coil transition. Surprisingly, force quench does not alter the nature of the refolding kinetics, but merely increases the height of the free-energy folding barrier. Force quench refolding times scale as tau(F) approximately tau(F)(0)exp(f(q)Deltax(f)/k(B)T), where Deltax(f) approximately 0.6 nm is the location of the average transition state along the reaction coordinate given by end-to-end distance. We predict that tau(F) and the folding mechanism can be dramatically altered by the initial and/or final values of force. The implications of our results for design and analysis of experiments are discussed. PMID- 16373510 TI - Internalization and phagosome escape required for Francisella to induce human monocyte IL-1beta processing and release. AB - Macrophage responses to Francisella infection have been characterized previously by subdued proinflammatory responses; however, these studies have generally focused on macrophage cell lines or monocyte-derived macrophages. Therefore, we studied the ability of fresh human blood monocytes to engulf and respond to Francisella by using the live vaccine strain variant and Francisella novicida. Because Francisella organisms have been reported to escape from the phagolysosome into the cytosol, we hypothesized that this escape may trigger the activation of caspase-1. Francisella tularensis variants were readily taken up by fresh human CD14(+) monocytes, inducing the release of IL-1beta, as well as IL-8, in a time- and dose-dependent fashion. Importantly, whereas live and dead Escherichia coli, F. novicida, and live vaccine strain, as well as the LPS of E. coli, were able to induce abundant IL-1beta mRNA synthesis and intracellular pro-IL-1beta production, only live Francisella induced enhanced IL-1beta processing and release (51 +/- 10 vs. 7.1 +/- 2.1 ng/ml, for F. novicida vs. E. coli LPS; P = 0.0032). Cytochalasin D blocked the Francisella internalization and the Francisella-induced monocyte IL-1beta processing and release but not that induced by the exogenous stimulus E. coli LPS. Also, killing bacteria did not block uptake but significantly diminished the IL-1beta processing and release that was induced by Francisella. Blocking bacterial escape from the phagosome into the cytosol also decreased IL-1beta but not IL-8 release. These findings demonstrate that Francisella organisms efficiently induce IL-1beta processing and release in fresh monocytes by means of a sensing system that requires the uptake of live bacteria capable of phagosome escape. PMID- 16373513 TI - Medical professionalism and ideological symbols in doctors' rooms. PMID- 16373512 TI - Protein fucosylation regulates synapsin Ia/Ib expression and neuronal morphology in primary hippocampal neurons. AB - Although fucose-alpha(1-2)-galactose [Fucalpha(1-2)Gal] carbohydrates have been implicated in cognitive processes such as long-term memory, the molecular mechanisms by which these sugars influence neuronal communication are not well understood. Here, we present molecular insights into the functions of Fucalpha(1 2)Gal sugars, demonstrating that they play a role in the regulation of synaptic proteins and neuronal morphology. We show that synapsins Ia and Ib, synapse specific proteins involved in neurotransmitter release and synaptogenesis, are the major Fucalpha(1-2)Gal glycoproteins in mature cultured neurons and the adult rat hippocampus. Fucosylation has profound effects on the expression and turnover of synapsin in cells and protects synapsin from degradation by the calcium activated protease calpain. Our studies suggest that defucosylation of synapsin has critical consequences for neuronal growth and morphology, leading to stunted neurite outgrowth and delayed synapse formation. We also demonstrate that Fucalpha(1-2)Gal carbohydrates are not limited to synapsin but are found on additional glycoproteins involved in modulating neuronal architecture. Together, our studies identify important roles for Fucalpha(1-2)Gal sugars in the regulation of neuronal proteins and morphological changes that may underlie synaptic plasticity. PMID- 16373514 TI - Choosing Health and the inner citadel. AB - It is argued in this paper that the latest UK government white paper on public health, Choosing Health, is vulnerable to a charge of paternalism. For some years libertarians have levelled this charge at public health policies. The white paper tries to avoid it by constant reference to informed choice and choice related terms. The implication is that the government aims only to inform the public of health issues; how they respond is up to them. It is argued here, however, that underlying the notion of informed choice is a Kantian, "inner citadel" view of autonomy. According to this view, each of us acts autonomously only when we act in accord with reason. On such a view it is possible to justify coercing, cajoling, and conning people on the basis that their current behaviour is not autonomous because it is subject to forces that cause irrational choice, such as addiction. "Informed choice" in this sense is compatible with paternalism. This paternalism can be seen in public health policies such as deceptive advertising and the treatment of "bad habits" as addictions. Libertarians are bound to object to this. In the concluding section, however, it is suggested that public health can, nonetheless, find ethical succour from alternative approaches. PMID- 16373515 TI - What should we say? AB - Abstract ethics mostly focuses on what we do. One form of action is a speech act. What we say can have profound effects. We can and should choose our words and how we speak wisely. When someone close to us suffers an injury or serious illness, a duty of beneficence requires that we support that person through beneficial words or actions. Though our intentions are most often benign, by what we say we often make the unfortunate person feel worse. Beginning with two personal accounts, this article explains what can go wrong in the compassionate speech of wellwishers, and uncovers some of the reasons why people say things that are hurtful or harmful. Despite a large body of clinical evidence, there is no perfect strategy for comforting a friend or relative who is ill, and sometimes even the best thing to say can still be perceived as insensitive and hurtful. In some cases, we may have good reason to knowingly say a hurtful or insensitive thing. Saying these 'wrong' things can sometimes be the best way to help a person in the long term. To complicate matters, there can be moral reasons for overriding what is good for the patient. What kind of admonishments should we make to a badly behaved patient? What is the value of authenticity in our communication with the people we love? These questions demand an ethical defence of those speech acts which are painful to hear but which need to be said, and of those which go wrong despite the best efforts of the wellwisher. We offer an ethical account, identifying permissible and impermissible justifications for the things we say to a person with a serious injury or illness. PMID- 16373516 TI - Body art and medical need. AB - A company called Biojewelry has proposed to take a sample of bone tissue from a couple and to grow this sample into wedding rings. One of the ethical problems that such a proposal faces is that it implies surgery without medical need. To this end, only couples with a prior need for surgery are being considered. This paper examines the question of whether such a stipulation is necessary. It is suggested that, though medical need and the provision of health and wellbeing is overwhelmingly the warrant for surgical intervention, there is no reason in principle why other, non-medical, projects such as jewelry creation might not also warrant surgical intervention. Implicitly, this line of thought forces us to consider the proper place of surgical intervention--that is, to ask what surgeons are for. PMID- 16373517 TI - Bioethics and health and human rights: a critical view. AB - Recent decades have seen the emergence of two new fields of inquiry into ethical issues in medicine. These are the fields of bioethics and of health and human rights. In this critical review of these fields, the author argues that bioethics, partly because it has been construed so broadly, suffers from quality control problems. The author also argues that the field of health and human rights is superfluous because it does nothing that cannot be done by either bioethics of the law. PMID- 16373518 TI - Should children's autonomy be respected by telling them of their imminent death? AB - Respect for an individual's autonomy determines that doctors should inform patients if their illness is terminal. This becomes complicated when the terminal diagnosis is recent and death is imminent. The authors examine the admission to paediatric intensive care of an adolescent with terminal respiratory failure. While fully ventilated, the patient was kept sedated and comfortable but when breathing spontaneously he was capable of non-verbal communication and understanding. Once resedated and reintubated, intense debate ensued over whether to wake the patient to tell him he was going to die. The authors discuss the ethical arguments that surrounded their decision. PMID- 16373519 TI - Commentary: Should children's autonomy be respected by telling them of their imminent death? PMID- 16373520 TI - A profession selling out: lamenting the paradigm shift in physician advertising. PMID- 16373521 TI - Mr Marty's muddle: a superficial and selective case for euthanasia in Europe. AB - In April 2004 the Parliamentary Assembly of the Council of Europe debated a report from its Social, Health and Family Affairs Committee (the Marty Report), which questioned the Council of Europe's opposition to legalising euthanasia. This article exposes the Report's flaws, not least its superficiality and selectivity. PMID- 16373522 TI - Commentary: Euthanasia in Europe: a critique of the Marty's report. PMID- 16373524 TI - Informing research participants of research results: analysis of Canadian university based research ethics board policies. AB - BACKGROUND: Despite potential benefits of the return of research results to research participants, the TriCouncil Policy Statement (TCPS), which reflects Canadian regulatory ethical requirements, does not require this. The policies of Canadian research ethics boards (REBs) are unknown. OBJECTIVES: To examine the policies of Canadian university based REBs regarding returning results to research participants, and to ascertain if the presence/absence of a policy may be influenced by REB member composition. DESIGN: Email survey of the coordinators of Canadian university based REBs to determine the presence/absence of a policy on return of research results to research participants both during an ongoing study and at conclusion. REB coordinators were asked to return a copy of the policy or guidelines and to describe the member composition of their REB. FINDINGS: Of 50 REBs that were contacted 34 (68%) responded and 22 (64.7%) met the inclusion criteria. Two (9.1%) had a policy that governed the return of research results while on a study, and seven (31.8%) following the completion of a study. Presence of an ethicist or a lawyer on the REB did not influence the presence/absence of such policies. No REBs had specific guidelines describing how participants should be informed of results. CONCLUSIONS: Most REBs did not require researchers to disclose study results to research participants either during or following a study. Thus this study identifies an ethical shortcoming in the conduct of human research in Canada. It has also demonstrated that there are no clear recommendations by REBs to facilitate the return of results to participants following research projects. PMID- 16373523 TI - What's in a name? Embryos, entities, and ANTities in the stem cell debate. AB - This paper discusses two proposals to the US President's Council on Bioethics that try to overcome the issue of killing embryos in embryonic stem (ES) cell research and argues that neither of them can hold good as a compromise solution. The author argues that (1) the groups of people for which the compromises are intended neither need nor want the two compromises, (2) the US government and other governments of countries with restrictive regulation on ES cell research have not provided a clear and sound justification to take into account minority views on the protection of human life to such a considerable extent as to constrain the freedom of research in the area of stem cell research, and (3) the best way to deal with these issues is to accept that many people and most governments adopt a gradualist and variable viewpoint on the human embryo which implies that embryos can be sacrificed for good reasons and to try to find other, less constraining, ways to take into account minority views on the embryo. Finally, another more efficient and time and money sparing compromise will be proposed for those who accept IVF, a majority in most societies. PMID- 16373525 TI - Equitable treatment for HIV/AIDS clinical trial participants: a focus group study of patients, clinician researchers, and administrators in Western Kenya. AB - OBJECTIVES: To describe the concerns and priorities of key stakeholders in a developing country regarding ethical obligations held by researchers and perceptions of equity or "what is fair" for study participants in an HIV/AIDS clinical drug trial. DESIGN: Qualitative study with focus groups. SETTING: Teaching and referral hospital and rural health centre in Western Kenya. PARTICIPANTS: Potential HIV/AIDS clinical trial participants, clinician researchers, and administrators. RESULTS: Eighty nine individuals participated in a total of 11 focus groups over a four month period. The desire for continued drug therapy, most often life long, following an HIV/AIDS clinical trial was the most common priority expressed in all focus groups. Patients with and without HIV/AIDS also thought subsidizing of drug therapies and education were critical forms of compensation for clinical trial participation. Financial incentives were considered important primarily for purchasing drug therapy as well as obtaining food. Patients noted a concern for the potential mismanagement of any money offered. Clinician researchers and administrators felt strongly that researchers have a moral obligation to participants following a trial to provide continued drug therapy, adverse event monitoring, and primary care. Finally, clinician researchers and administrators stressed the need for thorough informed consent to avoid coercion of study participants. CONCLUSIONS: Kenyan patients, clinician researchers, and administrators believe that it would be unfair to stop antiretroviral therapy following an HIV/AIDS clinical trial and that researchers have a long term obligation to participants. PMID- 16373526 TI - Brief report on the experience of using proxy consent for incapacitated adults. AB - The Medicines for Human Use (Clinical Trials) Regulations 2004, which came into force in the UK in May 2004, cover the conduct of clinical trials on medicinal products. They allow a legal representative (a person not connected with the conduct of the trial) to consent to the participation of incompetent adults in medical research. Currently, very little is known about how such representatives will make their decisions. We have experience with proxy consent for older adults in a large, national trial. From 2445 potentially eligible but incapacitated patients, proxy, relative assent resulted in trial participation of only 87 (3.6%) patients. The reasons for this were that a large number of incapacitated patients had no relative available for assent (2286), but also a high proportion of relatives approached refused to provide assent (72/159, 45.3%). In comparison, 17.7% of patients declined participation in the trial.Proxy consent allowed only a small increase in trial recruitment of incapacitated patients. The fact that a greater proportion of relatives than patients refused to provide assent implies that they were more cautious than the patients themselves, or perhaps used different criteria, when making their decision. In future research involving incapacitated older patients there is likely to be heavy reliance on proxy consent provision by legal representatives. Our findings imply that consent decisions of legal representatives will not necessarily reflect those of patients themselves. PMID- 16373527 TI - The predictive role of blood glucose for mortality in subjects with cardiovascular disease. AB - Using the Framingham Heart Study data (United States, 1948-1978), the authors examined the association of blood glucose with 2-year all-cause, cardiovascular, and noncardiovascular mortality in subjects with documented cardiovascular disease. After adjustment for systolic blood pressure, cholesterol, body mass index, cigarette smoking, and use of antihypertensive agents, they found that glucose was a strong, independent predictor of mortality. However, the relations for men and women were qualitatively different. For men, adjusted mortality risk increased very rapidly through the normal range (from 4.12% at 3.89 mmol/liter (70 mg/dl) to 12.26% at 5.55 mmol/liter (100 mg/dl)) and was flat at 12.26% thereafter. For women, risk was flat at 3.65% through the normal range and then increased rapidly, reaching 8.34% at 6.99 mmol/liter (126 mg/d), but increased much more slowly thereafter. Exactly analogous relations held for cardiovascular mortality. For men and women combined, noncardiovascular mortality increased from 1.82% at 3.89 mmol/liter to 2.06% at 5.55 mmol/liter to 2.29% at 6.99 mmol/liter (p for trend = 0.009). These findings suggest that although 5.55 mmol/liter (normal) may be a useful mortality risk division (albeit with different implications for the two sexes), 6.99 mmol/liter (diabetic) is not, especially for men. PMID- 16373528 TI - Resistance to the antineoplastic agent gallium nitrate results in marked alterations in intracellular iron and gallium trafficking: identification of novel intermediates. AB - Gallium (Ga) shows significant antitumor activity by markedly interfering with iron (Fe) metabolism, and (67)Ga is used as a radio-imaging agent for cancer detection. Therefore, the mechanisms involved in (67)Ga uptake, metabolism, and resistance are critical to understand. The development of tumor lines that are gallium-resistant suggests (67)Ga uptake may be different in these cells, providing an opportunity for understanding intracellular (67)Ga and (59)Fe transport and gallium resistance. In this study, gallium-resistant cells were used to assess (67)Ga and (59)Fe uptake using native polyacrylamide gel electrophoresis autoradiography. In contrast to the common view that (67)Ga and (59)Fe use the same uptake pathways, we show that the trafficking of these two metal ions is different in cells either resistant (R) or sensitive (S) to gallium. Indeed, in contrast to (59)Fe, little (67)Ga is incorporated into ferritin, with most present as a labile (67)Ga pool. We also report unique changes in (67)Ga and (59)Fe trafficking between R and S cells. In particular, in R cells, there was a distinct transferrin-transferrin receptor 1-hemochromatosis protein (HFE) complex (band B) not observed in S cells. Furthermore, because HFE regulates iron and gallium uptake, the two Tf-TfR1-HFE complexes in R cells may be involved in reduced (67)Ga and (59)Fe uptake compared with S cells. In S cells, a novel iron-binding intermediate (band D) was identified that was not present in R cells and may be a "sensitivity factor" to gallium. In contrast to the general view that (67)Ga and (59)Fe use the same or similar uptake pathways, we show that their distribution and trafficking is markedly different in R and S cells. PMID- 16373529 TI - S-adenosyl-L-methionine attenuates hepatotoxicity induced by agonistic Jo2 Fas antibody following CYP2E1 induction in mice. AB - S-Adenosyl-l-methionine (SAM) has been shown to be hepatoprotective against many toxic agents. Its possible effectiveness in protecting against CYP2E1-dependent toxicity is not known. We recently reported that treatment of mice with pyrazole to induce CYP2E1 increased hepatotoxicity produced by Fas agonistic Jo2 antibody. The current study was designed to investigate the effect of exogenous administration of SAM on the synergistic hepatotoxicity produced by Fas agonistic Jo2 antibody plus CYP2E1 following pyrazole pretreatment in C57BL/6 mice. Suboptimal administration of Jo2 Fas antibody combined with pyrazole pretreatment caused severe hepatotoxicity as determined by elevations in serum transaminase levels and histopathology. Exogenous administration of SAM (50 mg i.p./kg body weight every 12 h for 3 days) significantly decreased serum transaminases and ameliorated morphological changes of the liver. Addition of SAM elevated hepatic SAM and total reduced glutathione levels and inhibited CYP2E1 activity. SAM also lowered the elevated oxidative stress (lipid peroxidation, protein carbonyls, and superoxide production) and nitrosative stress (induction of inducible nitric oxide synthase and 3-nitrotyrosine adducts) and increases in caspase-8 and -3 activation produced by the pyrazole plus Jo2 treatment. SAM did not prevent the increase in serum TNF-alpha levels or the decrease in catalase activity in this model. These results indicate that SAM can have an important hepatoprotective role as an effective reagent against Fas plus CYP2E1-induced hepatotoxicity by lowering oxidative and nitrosative stress. PMID- 16373530 TI - In vitro and in vivo anti-inflammatory activity of a seed preparation containing phenethylisothiocyanate. AB - Winter cress (Barbarea verna) seed preparations rich in phenethylisothiocyanate (PEITC) had strong in vivo and in vitro anti-inflammatory activity, significantly reducing the size of carrageenan-induced rat paw edema. This in vivo effect was comparable with that of the nonsteroidal anti-inflammatory drug aspirin. The seed preparation, in a concentration-dependent manner, reduced the mRNA levels of inflammation-related genes such as the inducible forms of cyclooxygenase and nitric-oxide synthase and the proinflammatory cytokine interleukin in lipopolysaccharide-stimulated mouse macrophage cell line RAW 264.7. Activity of the seed preparation was similar to that of the synthetic PEITC. PEITC was the most active of five different forms of isothiocyanate tested for their effects on in vitro proinflammatory gene expression. In vitro activity of the seed preparation was also compared with that of two known anti-inflammatory drugs. We conclude that Barbarea verna seed preparation may function as a potent anti inflammatory agent, interfering with the transcription of proinflammatory genes. PMID- 16373531 TI - Editorial expression of concern. PMID- 16373532 TI - Magnetosomes are cell membrane invaginations organized by the actin-like protein MamK. AB - Magnetosomes are membranous bacterial organelles sharing many features of eukaryotic organelles. Using electron cryotomography, we found that magnetosomes are invaginations of the cell membrane flanked by a network of cytoskeletal filaments. The filaments appeared to be composed of MamK, a homolog of the bacterial actin-like protein MreB, which formed filaments in vivo. In a mamK deletion strain, the magnetosome-associated cytoskeleton was absent and individual magnetosomes were no longer organized into chains. Thus, it seems that prokaryotes can use cytoskeletal filaments to position organelles within the cell. PMID- 16373533 TI - The second ring-moon system of Uranus: discovery and dynamics. AB - Deep exposures of Uranus taken with the Hubble Space Telescope reveal two small moons and two faint rings. All of them orbit outside of Uranus's previously known (main) ring system but are interior to the large, classical moons. The outer new moon, U XXVI Mab, orbits at roughly twice the radius of the main rings and shares its orbit with a dust ring. The second moon, U XXVII Cupid, orbits just interior to the satellite Belinda. A second ring falls between the orbits of Portia and Rosalind, in a region with no known source bodies. Collectively, these constitute a densely packed, rapidly varying, and possibly unstable dynamical system. PMID- 16373534 TI - Pairing and phase separation in a polarized Fermi gas. AB - We report the observation of pairing in a gas of atomic fermions with unequal numbers of two components. Beyond a critical polarization, the gas separates into a phase that is consistent with a superfluid paired core surrounded by a shell of normal unpaired fermions. The critical polarization diminishes with decreasing attractive interaction. For near-zero polarization, we measured the parameter beta = -0.54 +/- 0.05, describing the universal energy of a strongly interacting paired Fermi gas, and found good agreement with recent theory. These results are relevant to predictions of exotic new phases of quark matter and of strongly magnetized superconductors. PMID- 16373535 TI - Fermionic superfluidity with imbalanced spin populations. AB - We established superfluidity in a two-state mixture of ultracold fermionic atoms with imbalanced state populations. This study relates to the long-standing debate about the nature of the superfluid state in Fermi systems. Indicators for superfluidity were condensates of fermion pairs and vortices in rotating clouds. For strong interactions, near a Feshbach resonance, superfluidity was observed for a broad range of population imbalances. We mapped out the superfluid regime as a function of interaction strength and population imbalance and characterized the quantum phase transition to the normal state, known as the Pauli limit of superfluidity. PMID- 16373536 TI - A bacterial inhibitor of host programmed cell death defenses is an E3 ubiquitin ligase. AB - The Pseudomonas syringae protein AvrPtoB is translocated into plant cells, where it inhibits immunity-associated programmed cell death (PCD). The structure of a C terminal domain of AvrPtoB that is essential for anti-PCD activity reveals an unexpected homology to the U-box and RING-finger components of eukaryotic E3 ubiquitin ligases, and we show that AvrPtoB has ubiquitin ligase activity. Mutation of conserved residues involved in the binding of E2 ubiquitin conjugating enzymes abolishes this activity in vitro, as well as anti-PCD activity in tomato leaves, which dramatically decreases virulence. These results show that Pseudomonas syringae uses a mimic of host E3 ubiquitin ligases to inactivate plant defenses. PMID- 16373537 TI - Breakthrough of the year. PMID- 16373538 TI - Breakthrough of the year: evolution in action. PMID- 16373540 TI - Breakthrough of the year: scorecard 2004. PMID- 16373539 TI - Breakthrough of the year: the runners-up. PMID- 16373541 TI - Breakdown of the year: U.S. particle physics. PMID- 16373542 TI - Disasters: searching for lessons from a bad year. PMID- 16373543 TI - Breakthrough of the year: areas to watch in 2006. PMID- 16373544 TI - Stem cells. Cloning researcher says work is flawed but claims results stand. PMID- 16373545 TI - Ancient DNA. New methods yield mammoth samples. PMID- 16373546 TI - Drug testing. Massive trial of Celebrex seeks to settle safety concerns. PMID- 16373548 TI - U.S. science policy. Bill seeks billions to bolster research. PMID- 16373547 TI - Deep-sea drilling. Scientific drill ship to be reborn. PMID- 16373549 TI - Condensed-matter physics. Mismatched cold atoms hint at a stellar new superfluid. PMID- 16373550 TI - Immunology. Jawless fish have form of adaptive immunity. PMID- 16373551 TI - Satellite navigation. Europe's answer to GPS could be a boon for research. PMID- 16373552 TI - Gene therapy. Putting the fingers on gene repair. PMID- 16373553 TI - Tree growth. The sky is not the limit. PMID- 16373554 TI - Meeting: American Geophysical Union. San Andreas drillers find a strangely weak fault. PMID- 16373556 TI - Meeting: American Geophysical Union. Mars saucer mystery baffles the experts. PMID- 16373555 TI - Meeting: American Geophysical Union. An early, muddy Mars just right for life. PMID- 16373557 TI - Meeting: American Geophysical Union. Snapshots from the meeting. PMID- 16373558 TI - Retraction. PMID- 16373560 TI - Highlighting the STAR collaboration. PMID- 16373559 TI - Inka accounting practices. PMID- 16373561 TI - Communication. Social values and the governance of science. PMID- 16373562 TI - Evolution. Is the "Big Bang" in animal evolution real? PMID- 16373563 TI - Physiology. The tick-tock of aging? PMID- 16373564 TI - Chemistry. Nuclear spin conversion in molecules. PMID- 16373565 TI - Geochemistry. A tale of early Earth told in zircons. PMID- 16373566 TI - Retrospective: Richard E. Smalley (1943-2005). PMID- 16373567 TI - Appendage regeneration in adult vertebrates and implications for regenerative medicine. AB - The regeneration of complex structures in adult salamanders depends on mechanisms that offer pointers for regenerative medicine. These include the plasticity of differentiated cells and the retention in regenerative cells of local cues such as positional identity. Limb regeneration proceeds by the local formation of a blastema, a growth zone of mesenchymal stem cells on the stump. The blastema can regenerate autonomously as a self-organizing system over variable linear dimensions. Here we consider the prospects for limb regeneration in mammals from this viewpoint. PMID- 16373568 TI - Torus-margo pits help conifers compete with angiosperms. AB - The unicellular conifer tracheid should have greater flow resistance per length (resistivity) than the multicellular angiosperm vessel, because its high resistance end-walls are closer together. However, tracheids and vessels had comparable resistivities for the same diameter, despite tracheids being over 10 times shorter. End-wall pits of tracheids averaged 59 times lower flow resistance on an area basis than vessel pits, owing to the unique torus-margo structure of the conifer pit membrane. The evolution of this membrane was as hydraulically important as that of vessels. Without their specialized pits, conifers would have 38 times the flow resistance, making conifer-dominated ecosystems improbable in an angiosperm world. PMID- 16373569 TI - Animal evolution and the molecular signature of radiations compressed in time. AB - The phylogenetic relationships among most metazoan phyla remain uncertain. We obtained large numbers of gene sequences from metazoans, including key understudied taxa. Despite the amount of data and breadth of taxa analyzed, relationships among most metazoan phyla remained unresolved. In contrast, the same genes robustly resolved phylogenetic relationships within a major clade of Fungi of approximately the same age as the Metazoa. The differences in resolution within the two kingdoms suggest that the early history of metazoans was a radiation compressed in time, a finding that is in agreement with paleontological inferences. Furthermore, simulation analyses as well as studies of other radiations in deep time indicate that, given adequate sequence data, the lack of resolution in phylogenetic trees is a signature of closely spaced series of cladogenetic events. PMID- 16373570 TI - Separation and conversion dynamics of four nuclear spin isomers of ethylene. AB - Molecules with three or more nuclei of nonzero spin exist as discrete spin isomers whose interconversion in the gas phase is generally considered improbable. We have studied the interconversion process in ethylene by creating a sample depleted in the B2u nuclear spin isomer. The separation was achieved through spatial drift of this isomer induced by resonant absorption of narrow band infrared light. Evolution of the depleted sample revealed conversion between B2u and B3u isomers at a rate linearly proportional to pressure, with a rate constant of 5.5 (+/-0.8) x 10(-4) s(-1) torr(-1). However, almost no change was observed in the Ag isomer populations. The results suggest a spin conversion mechanism in C2H4 via quantum relaxation within the same inversion symmetry. PMID- 16373571 TI - Synthesis of imido analogs of the uranyl ion. AB - Here we describe the synthesis of two imido analogs of the uranyl ion, UO(2+)2, in which the oxygens are replaced by divalent alkyl or aryl nitrogen groups: U(NtBu)2I2(THF)2 (1) and U(NPh)2I2(THF)3 (2) (where tBu is tert-butyl and THF is tetrahydrofuran). Both compounds have been fully characterized by standard analytical techniques, including x-ray crystallography, and the chemical bonding between the metal center and the nitrogen ligands was quantified by using hybrid density functional theory calculations. As expected for a uranyl analog, these complexes exhibit linear N-U-N linkages and very short U-N bonds. In addition, the theoretical calculations show strong involvement of the 5f and 6d electrons in the U-N bonding. PMID- 16373572 TI - Trading water for carbon with biological carbon sequestration. AB - Carbon sequestration strategies highlight tree plantations without considering their full environmental consequences. We combined field research, synthesis of more than 600 observations, and climate and economic modeling to document substantial losses in stream flow, and increased soil salinization and acidification, with afforestation. Plantations decreased stream flow by 227 millimeters per year globally (52%), with 13% of streams drying completely for at least 1 year. Regional modeling of U.S. plantation scenarios suggests that climate feedbacks are unlikely to offset such water losses and could exacerbate them. Plantations can help control groundwater recharge and upwelling but reduce stream flow and salinize and acidify some soils. PMID- 16373573 TI - X-ray structure of the EmrE multidrug transporter in complex with a substrate. AB - EmrE is a prototype of the Small Multidrug Resistance family of efflux transporters and actively expels positively charged hydrophobic drugs across the inner membrane of Escherichia coli. Here, we report the x-ray crystal structure, at 3.7 angstrom resolution, of one conformational state of the EmrE transporter in complex with a translocation substrate, tetraphenylphosphonium. Two EmrE polypeptides form a homodimeric transporter that binds substrate at the dimerization interface. The two subunits have opposite orientations in the membrane and adopt slightly different folds, forming an asymmetric antiparallel dimer. This unusual architecture likely confers unidirectionality to transport by creating an asymmetric substrate translocation pathway. On the basis of available structural data, we propose a model for the proton-dependent drug efflux mechanism of EmrE. PMID- 16373574 TI - A developmental timing microRNA and its target regulate life span in C. elegans. AB - The microRNA lin-4 and its target, the putative transcription factor lin-14, control the timing of larval development in Caenorhabditis elegans. Here, we report that lin-4 and lin-14 also regulate life span in the adult. Reducing the activity of lin-4 shortened life span and accelerated tissue aging, whereas overexpressing lin-4 or reducing the activity of lin-14 extended life span. Lifespan extension conferred by a reduction in lin-14 was dependent on the DAF-16 and HSF-1 transcription factors, suggesting that the lin-4-lin-14 pair affects life span through the insulin/insulin-like growth factor-1 pathway. This work reveals a role for microRNAs and developmental timing genes in life-span regulation. PMID- 16373575 TI - fgf20 is essential for initiating zebrafish fin regeneration. AB - Epimorphic regeneration requires the presence or creation of pluripotent cells capable of reproducing lost organs. Zebrafish fin regeneration is mediated by the creation of blastema cells. Here, we characterize the devoid of blastema (dob) mutant that fails fin regeneration during initial steps, forms abnormal regeneration epithelium, and does not form blastema. This mutation has no impact on embryonic survival. Dob results from an fgf20a null mutation, Y148S. Fgf20a is expressed during initiation of fin regeneration at the epithelial-mesenchymal boundary and later overlaps with the blastema marker msxb. Thus, fgf20a has a regeneration-specific requirement, initiating fin regeneration, and controlling blastema formation. PMID- 16373576 TI - Protein synthesis upon acute nutrient restriction relies on proteasome function. AB - The mechanisms that protect mammalian cells against amino acid deprivation are only partially understood. We found that during an acute decrease in external amino acid supply, before up-regulation of the autophagosomal-lysosomal pathway, efficient translation was ensured by proteasomal protein degradation. Amino acids for the synthesis of new proteins were supplied by the degradation of preexisting proteins, whereas nascent and newly formed polypeptides remained largely protected from proteolysis. Proteasome inhibition during nutrient deprivation caused rapid amino acid depletion and marked impairment of translation. Thus, the proteasome plays a crucial role in cell survival after acute disruption of amino acid supply. PMID- 16373577 TI - Category-specific cortical activity precedes retrieval during memory search. AB - Here we describe a functional magnetic resonance imaging study of humans engaged in memory search during a free recall task. Patterns of cortical activity associated with the study of three categories of pictures (faces, locations, and objects) were identified by a pattern-classification algorithm. The algorithm was used to track the reappearance of these activity patterns during the recall period. The reappearance of a given category's activity pattern correlates with verbal recalls made from that category and precedes the recall event by several seconds. This result is consistent with the hypothesis that category-specific activity is cueing the memory system to retrieve studied items. PMID- 16373578 TI - Inducible nitric oxide synthase binds, S-nitrosylates, and activates cyclooxygenase-2. AB - Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are two major inflammatory mediators. Here we show that iNOS specifically binds to COX-2 and S nitrosylates it, enhancing COX-2 catalytic activity. Selectively disrupting iNOS COX-2 binding prevented NO-mediated activation of COX-2. This synergistic molecular interaction between two inflammatory systems may inform the development of anti-inflammatory drugs. PMID- 16373580 TI - Vascular adrenergic tone and structural narrowing constrain reactive hyperemia in skeletal muscle of obese Zucker rats. AB - Previous studies have demonstrated that skeletal muscle perfusion is impaired in obese Zucker rats (OZR) under control conditions and with elevated metabolic demand versus responses in lean Zucker rats (LZR). To further our understanding of processes contributing to impaired perfusion, we determined whether hyperemic responses following periods of occlusion were altered in skeletal muscle of OZR versus LZR. In isolated hindlimbs, basal blood flow in OZR was less than in LZR, and total perfusion responses after 30, 90, and 180 s of occlusion were reduced. Treatment of animals with an antioxidant (polythethylene glycol-superoxide dismutase) had no effect on reactive hyperemia, although blockade of alpha adrenoreceptors (alpha1 > alpha2) improved responses to 30 and 90 s of occlusion; responses to 180 s of occlusion were unaltered. Pump perfusion of a dilated distal hindlimb demonstrated that increased volume flow elicited a greater increase in perfusion pressure in OZR versus LZR, suggesting structural contributions to an increased vascular resistance. Responses were comparable for in situ cremaster muscle because reactive hyperemia following serial arteriolar occlusion was attenuated in OZR versus LZR, treatment with polythethylene glycol superoxide dismutase was ineffective, and hyperemic responses were improved following inhibition of alpha-adrenoreceptors (alpha1 > alpha2). Treatment of cremaster muscle with adenosine (10(-3) M) caused flow to increase to a level comparable to that following 180 s of occlusion in both strains, although this level was reduced in OZR versus LZR. These results suggest that increased adrenergic tone may constrain reactive hyperemia in OZR with brief occlusion, although structural increases in vascular resistance can contribute to constrained perfusion after longer periods of occlusion. PMID- 16373579 TI - Diversity and function of adaptive immune receptors in a jawless vertebrate. AB - Instead of the immunoglobulin-type antigen receptors of jawed vertebrates, jawless fish have variable lymphocyte receptors (VLRs), which consist of leucine rich repeat (LRR) modules. Somatic diversification of the VLR gene is shown here to occur through a multistep assembly of LRR modules randomly selected from a large bank of flanking cassettes. The predicted concave surface of the VLR is lined with hypervariable positively selected residues, and computational analysis suggests a repertoire of about 10(14) unique receptors. Lamprey immunized with anthrax spores responded with the production of soluble antigen-specific VLRs. These findings reveal that two strikingly different modes of antigen recognition through rearranged lymphocyte receptors have evolved in the jawless and jawed vertebrates. PMID- 16373581 TI - Inositol phospholipids localized to caveolae in rat heart are regulated by alpha1 adrenergic receptors and by ischemia-reperfusion. AB - Postischemic reperfusion of rat or mouse hearts causes generation of inositol (1,4,5)trisphosphate [Ins(1,4,5)P3] and the initiation of arrhythmias. In the current study we investigated the possibility that the enhanced Ins(1,4,5)P3 generation in postischemic reperfusion was associated with an increased availability of the precursor lipid phosphatidylinositol(4,5)bisphosphate (PIP2) for alpha1-adrenergic receptor-activated phospholipase C (PLC). Isolated-perfused rat hearts were labeled with [3H]inositol and subjected to ischemia-reperfusion or stimulation with norepinephrine under normoxic conditions. Caveolar fractions were prepared by buoyant density sucrose gradient centrifugation. [3H]PIP2 was concentrated in caveolae, along with Galphaq and PLCbeta1b. Caveolae contained only 27.3 +/- 6.9% (means +/- SE, n = 6) of the total alpha1-adrenergic receptor complement of the heart. These did not migrate to PIP2-containing caveolar fractions with norepinephrine stimulation under normoxic conditions, even though caveolar PIP2 was depleted. In contrast, [3H]PIP2 in caveolae increased during 2 min of reperfusion, independently of norepinephrine release and thus of alpha1 adrenergic receptor activation. The increased PIP2 in the caveolar fractions where signaling proteins are concentrated may be critical for the heightened generation of Ins(1,4,5)P3 in early reperfusion. PMID- 16373582 TI - Cardiac microimpedance measurement in two-dimensional models using multisite interstitial stimulation. AB - We analyzed central interstitial potential differences during multisite stimulation to assess the feasibility of using those recordings to measure cardiac microimpedances in multidimensional preparations. Because interstitial current injected and removed using electrodes with different proximities allows modulation of the portion of current crossing the membrane, we hypothesized that multisite interstitial stimulation would give rise to central interstitial potential differences that depend on intracellular and interstitial microimpedances, allowing measurement of those microimpedances. Simulations of multisite stimulation with fine and wide spacing in two-dimensional models that included dynamic membrane equations for guinea pig ventricular myocytes were performed to generate test data ( partial differentialphio). Isotropic interstitial and intracellular microimpedances were prescribed for one set of simulations, and anisotropic microimpedances with unequal ratios (intracellular to interstitial) along and across fibers were prescribed for another set of simulations. Microimpedance measurements were then obtained by making statistical comparisons between partial differentialphio values and interstitial potential differences from passive bidomain simulations (Deltaphio) in which a wide range of possible microimpedances were considered. Possible microimpedances were selected at 25% increments. After demonstrating the effectiveness of the overall method with microimpedance measurements using one-dimensional test data, we showed microimpedance measurements within 25% of prescribed values in isotropic and anisotropic models. Our findings suggest that development of microfabricated devices to implement the procedure would facilitate routine measurement as a component of cardiac electrophysiological study. PMID- 16373583 TI - C-phycocyanin protects against ischemia-reperfusion injury of heart through involvement of p38 MAPK and ERK signaling. AB - We previously showed that C-phycocyanin (PC), an antioxidant biliprotein pigment of Spirulina platensis (a blue-green alga), effectively inhibited doxorubicin induced oxidative stress and apoptosis in cardiomyocytes. Here we investigated the cardioprotective effect of PC against ischemia-reperfusion (I/R)-induced myocardial injury in an isolated perfused Langendorff heart model. Rat hearts were subjected to 30 min of global ischemia at 37 degrees C followed by 45 min of reperfusion. Hearts were perfused with PC (10 microM) or Spirulina preparation (SP, 50 mg/l) for 15 min before the onset of ischemia and throughout reperfusion. After 45 min of reperfusion, untreated (control) hearts showed a significant decrease in recovery of coronary flow (44%), left ventricular developed pressure (21%), and rate-pressure product (24%), an increase in release of lactate dehydrogenase and creatine kinase in coronary effluent, significant myocardial infarction (44% of risk area), and TdT-mediated dUTP nick end label-positive apoptotic cells compared with the preischemic state. PC or SP significantly enhanced recovery of heart function and decreased infarct size, attenuated lactate dehydrogenase and creatine kinase release, and suppressed I/R-induced free radical generation. PC reversed I/R-induced activation of p38 MAPK, Bax, and caspase-3, suppression of Bcl-2, and increase in TdT-mediated dUTP nick end label positive apoptotic cells. However, I/R also induced activation of ERK1/2, which was enhanced by PC treatment. Overall, these results for the first time showed that PC attenuated I/R-induced cardiac dysfunction through its antioxidant and antiapoptotic actions and modulation of p38 MAPK and ERK1/2. PMID- 16373584 TI - Contactless magnetocardiographic mapping in anesthetized Wistar rats: evidence of age-related changes of cardiac electrical activity. AB - Magnetocardiography (MCG) is the recording of the magnetic field (MF) generated by cardiac electrophysiological activity. Because it is a contactless method, MCG is ideal for noninvasive cardiac mapping of small experimental animals. The aim of this study was to assess age-related changes of cardiac intervals and ventricular repolarization (VR) maps in intact rats by means of MCG mapping. Twenty-four adult Wistar rats (12 male and 12 female) were studied, under anesthesia, with the same unshielded 36-channel MCG instrumentation used for clinical recordings. Two sets of measurements were obtained from each animal: 1) at 5 mo of age (297.5 +/- 21 g body wt) and 2) at 14 mo of age (516.8 +/- 180 g body wt). RR and PR intervals, QRS segment, and QTpeak, QTend, JTpeak, JTend, and Tpeak-end were measured from MCG waveforms. MCG imaging was automatically obtained as MF maps and as inverse localization of cardiac sources with equivalent current dipole and effective magnetic dipole models. After 300 s of continuous recording were averaged, the signal-to-noise ratio was adequate for study of atrial and ventricular MF maps and for three-dimensional localization of the underlying cardiac sources. Clear-cut age-related differences in VR duration were demonstrated by significantly longer QTend, JTend, and Tpeak-end in older Wistar rats. Reproducible multisite noninvasive cardiac mapping of anesthetized rats is simpler with MCG methodology than with ECG recording. In addition, MCG mapping provides new information based on quantitative analysis of MF and equivalent sources. In this study, statistically significant age-dependent variations in VR intervals were found. PMID- 16373585 TI - Aging and nonlinear heart rate control in a healthy population. AB - In recent years more studies are using nonlinear dynamics to describe cardiovascular control. Because of the large dispersion of physiological data, it is important to have large studies with both male and female participants to establish a range of physiological healthy values. This study investigated the effect of gender and age on nonlinear indexes. Nonlinear scaling properties were studied by using 1/f slope (where f is frequency), fractal dimension, and detrended fluctuation analysis short- and long-term correlations (DFAalpha(1) and DFAalpha(2), respectively). Nonlinear complexity was described with correlation dimension (CD), Lyapunov exponent (LE), and approximate entropy (ApEn). The population consisted of 135 women and 141 men (age, 18-71 yr). Twenty-four hour ECG recordings were obtained by using Holter monitoring. The recordings were split into daytime (8 AM-9 PM) and nighttime (11 PM-6 AM). A day-night variation was present in all nonlinear heart rate variability (HRV) indexes, except for the CD in the female population. During the night the percentage of CD values of surrogate data files differing from the CD value of the original data increased. All nonlinear indexes were significantly correlated with age. Deeper analysis per age category of 10 yr showed a stabilization in the age decline of the fractal dimension and ApEn at the age of > or =40 yr. The vagal pathways seemed to be more involved in the generation of nonlinear fluctuations. Higher nonlinear behavior was evident during the night. No clear difference between men and women was found in the nonlinear indexes. Nonlinear indexes decline with age. This can be related to the concept of decreasing autonomic modulation with advancing age. PMID- 16373586 TI - Calpains: a physiological regulator of the endothelial barrier? AB - The intracellular protease calpain, abundant in endothelial cells (EC), is assumed to be inactive under physiological conditions but may account for Ca2+ linked pathophysiological events. However, nonstimulated EC contained autolyzed, activated calpain. Adding 12-48 microM calpain inhibitor I (CI) or 0.5-1 microM of the novel, membrane-permeable conjugate of calpastatin peptide-penetratin (CPP) caused rapid rounding and retraction of cultured EC (phase contrast, capacitance) and translocation of Syk, Rac, and Rho to the membrane, signifying activation upon inhibition of calpain. Isolated hearts (guinea pig) perfused with 12 microM CI or 0.5 muM CPP developed coronary leak. We conclude that calpain is constitutively active in EC and regulates vascular permeability by governing cell cell attachment. PMID- 16373587 TI - Nuclear compartmentalization of FAK and FRNK in cardiac myocytes. AB - Focal adhesion kinase (FAK) and FAK-related non-kinase (FRNK) accumulate in the nucleus of cardiac myocytes during hypertensive hypertrophy. Nuclear FAK and FRNK are phosphorylated on different serines and form distinct bright spots. The subnuclear distribution of serine-phosphorylated FAK and FRNK was examined in this study by double labeling with fibrillarin, a component of nucleoli, and Sam68, a constituent of Sam68 nuclear bodies. We also investigated the role of protein kinase C (PKC)-mediated phosphorylation of FAK and FRNK on nuclear translocation. PKC activation by 12-O-tetradecanoylphorbol 13-acetate treatment increased serine phosphorylation of FAK and FRNK. Specifically, FAK was phosphorylated on serine 722 but not serine 910. On the other hand, FRNK was phosphorylated on serine 217, the equivalent site of FAK serine 910, but not serine 30, the homologous site of FAK serine 722. Serine-phosphorylated FAK and FRNK redistributed into the nucleus and formed distinct patterns. FAK with phosphorylation on serine 722 colocalized with Sam68 but not fibrillarin. On the contrary, FRNK phosphorylated on 217 coexisted with fibrillarin but not Sam68. Immunoprecipitation also confirmed that FAK associated with Sam68 and FRNK interacted with fibrillarin, respectively. These results suggest that FAK and FRNK target different nuclear subdomains by their association with distinct nuclear proteins. PMID- 16373588 TI - Changes in plasma protein extravasation in rat skin during inflammatory challenges evaluated by microdialysis. AB - Docetaxel and prostaglandin E1 (PGE1) increase transcapillary albumin extravasation and reduce interstitial fluid pressure in the skin. In this study the microdialysate concentration (Cm) of 125I-labeled human serum albumin (125I HSA) and different-sized endogenous plasma proteins (EPP) was compared to evaluate changes in transcapillary extravasation of plasma proteins. 125I-HSA was also used to estimate changes in the specific activity of albumin. Extravasation of 125I-HSA and EPP from plasma to interstitium in the rat skin was compared during continuous administration of docetaxel and PGE1 by using microdialysis in anesthetized rats. Also, 20 ml of Ringer solution (RS) were injected intravenously during 10 min in a separate group. Two hollow plasmapheresis fibers (3 cm, cut off 3,000 kDa), one acting as control, were placed subcutaneously on the back skin and perfused with RS (5 microl/min, 140 min, collected every 10 min). The size of the different EPP was estimated to be 73, 65, 56, 47, and 39 A, separated by a size-exclusion high-performance liquid chromatography column and quantified by UV detection (280 nm). Docetaxel (0.5 mg/ml, n = 5) increased Cm of 125I-HSA and EPP of sizes 73, 65, 56, and 39 A significantly (P < 0.05) compared with control. PGE1 (20 microg/ml, n = 6) increased Cm of 125I-HSA significantly (P < 0.05) but none of the different-sized EPP was increased compared with control. Intravenous RS (20 ml, n = 6) increased Cm of 125I-HSA and increased all the different-sized EPP significantly (P < 0.05) compared with control. Although the microdialysis method is able to monitor qualitative changes in capillary permeability, a quantitative determination of the capillary reflection coefficient or permeability-surface area product was not possible, because steady state between plasma and dialysate was not achieved during the measurement period. The different pattern of extravasation of EPP and 125I-HSA after docetaxel, PGE1, and RS indicates increased interstitial transport rate and/or increased capillary permeability after docetaxel and RS, whereas PGE1 seems to increase transcapillary fluid flux without altering the permeability. PMID- 16373589 TI - The circadian clock within the heart: potential influence on myocardial gene expression, metabolism, and function. AB - It is becoming increasingly clear that the intrinsic properties of both the heart and vasculature exhibit dramatic oscillations over the course of the day. Diurnal variations in the responsiveness of the cardiovascular system to environmental stimuli are mediated by a complex interplay between extracellular (i.e., neurohumoral factors) and intracellular (i.e., circadian clock) influences. The intracellular circadian clock is composed of a series of transcriptional modulators that together allow the cell to perceive the time of day, thereby enabling preparation for an anticipated stimulus. These molecular timepieces have been characterized recently within both vascular smooth muscle cells and cardiomyocytes, giving rise to a multitude of hypotheses relating to the potential role(s) of the circadian clock as a modulator of physiological and pathophysiological cardiovascular events. For example, evidence strongly supports the hypothesis that the circadian clock within the heart modulates myocardial metabolism, which in turn facilitates anticipation of diurnal variations in workload, substrate availability, and/or the energy supply-to-demand ratio. The purpose of this review is therefore to summarize our current understanding of the molecular events governing diurnal variations in the intrinsic properties of the heart, with special emphasis on the intramyocardial circadian clock. Whether impairment of this molecular mechanism contributes toward cardiovascular disease associated with hypertension, diabetes mellitus, shift work, sleep apnea, and/or obesity will be discussed. PMID- 16373590 TI - Cardiac endocrine function is an essential component of the homeostatic regulation network: physiological and clinical implications. AB - The discovery of cardiac natriuretic hormones required a profound revision of the concept of heart function. The heart should no longer be considered only as a pump but rather as a multifunctional and interactive organ that is part of a complex network and active component of the integrated systems of the body. In this review, we first consider the cross-talk between endocrine and contractile function of the heart. Then, based on the existing literature, we propose the hypothesis that cardiac endocrine function is an essential component of the integrated systems of the body and thus plays a pivotal role in fluid, electrolyte, and hemodynamic homeostasis. We highlight those studies indicating how alterations in cardiac endocrine function can better explain the pathophysiology of cardiovascular diseases and, in particular of heart failure, in which several target organs develop a resistance to the biological action of cardiac natriuretic peptides. Finally, we emphasize the concept that a complete knowledge of the cardiac endocrine function and of its relation with other neurohormonal regulatory systems of the body is crucial to correctly interpret changes in circulating natriuretic hormones, especially the brain natriuretic peptide. PMID- 16373591 TI - Diversion of blood flow from noncompliant to compliant vasculature in awake dogs: mechanical impact on right atrial pressure. AB - The distribution of cardiac output between compliant vasculature (e.g., splanchnic organs and skin) and noncompliant vasculature (e.g., skeletal muscle) is proposed to constitute an important determinant of the amount of blood available to the heart (central blood volume and pressure). The aim here was to directly test the hypothesis that diversion of blood flow from a relatively noncompliant vasculature (muscle) to compliant vasculature (splanchnic organs and skin) acts to reduce right atrial pressure. The approach was to inflate an occluder cuff on the terminal aorta for 30 s in one of two modes of ventricular pacing in five awake dogs with atrioventricular block and autonomic blockade. In one trial, cardiac output was maintained constant, meaning cuff inflation caused a portion of terminal aortic flow (a noncompliant circulation) to be diverted to the splanchnic and skin circulations (compliant circulations). In the other trial, arterial pressure was maintained constant, meaning blood flow to these other regions did not change. The response of right atrial pressure (corrected for differences in arterial pressure between the two trials) fit our hypothesis, being lower when blood flow was diverted to compliant regions. We conclude that a small (4% of cardiac output) diversion of blood flow from a noncompliant region to a compliant region reduces right atrial pressure by 0.7 mmHg. PMID- 16373592 TI - Calcium paradox of aldosteronism and the role of the parathyroid glands. AB - The hypercalciuria and hypermagnesuria that accompany aldosteronism contribute to a fall in plasma ionized extracellular Ca2+ and Mg2+ concentrations ([Ca2+]o and [Mg2+]o). Despite these losses and the decline in extracellular levels of these cations, total intracellular and cytosolic free Ca2+ concentration ([Ca2+]i) is increased and oxidative stress is induced. This involves diverse tissues, including peripheral blood mononuclear cells (PBMC) and plasma. The accompanying elevation in plasma parathyroid hormone (PTH) and reduction in bone mineral density caused by aldosterone (Aldo)-1% NaCl treatment (AldoST) led us to hypothesize that Ca2+ loading and altered redox state are due to secondary hyperparathyroidism (SHPT). Therefore, we studied the effects of total parathyroidectomy (PTx). In rats receiving AldoST, without or with a Ca2+ supplemented diet and/or PTx, we monitored urinary Ca2+ and Mg2+ excretion; plasma [Ca2+]o, [Mg2+]o, and PTH; PBMC [Ca2+]i and H2O2 production; plasma alpha1 antiproteinase activity; total Ca2+ and Mg2+ in bone, myocardium, and rectus femoris; and gp91(phox) labeling in the heart. We found that 1) the hypercalciuria and hypermagnesuria and decline (P < 0.05) in plasma [Ca2+]o and [Mg2+]o that occur with AldoST were not altered by the Ca2+-supplemented diet alone or with PTx; 2) the rise (P < 0.05) in plasma PTH with AldoST, with or without the Ca2+-supplemented diet, was prevented by PTx; 3) increased (P < 0.05) PBMC [Ca2+]i and H2O2 production, increased total Ca2+ in heart and skeletal muscle, and fall in bone Ca2+ and Mg2+ and plasma alpha1-antiproteinase activity with AldoST were abrogated (P < 0.05) by PTx; and 4) gp91(phox) activation in right and left ventricles at 4 wk of AldoST was attenuated by PTx. AldoST is accompanied by SHPT, with parathyroid gland-derived calcitropic hormones being responsible for Ca2+ overload in diverse tissues and induction of oxidative stress. SHPT plays a permissive role in the proinflammatory vascular phenotype. PMID- 16373593 TI - Effect of phosphodiesterase 5 inhibitor on alteration in vascular smooth muscle sensitivity and renal function in rats with liver cirrhosis. AB - Previous studies suggested that increased activity of phosphodiesterase (PDE)5 in the kidneys of cirrhotic rats contributes to sodium retention. This study examined the role of PDE5 in the changes in vascular reactivity, hemodynamics, and sodium excretion in rats with liver cirrhosis. Four weeks after bile duct ligation (BDL) or sham operation (SO), in vitro reactivity of aortic rings to various agents and in vivo effects of a PDE5-selective inhibitor [1,3-dimethyl-6 (2-propoxy-5-methanesulfonylamidophenyl)pyrazolo[3,4d]-pyrimidin-4-(5H)-one, DMPPO] were studied. The vasodilator responses to nitroglycerin and S-nitroso-N acetyl-penicillamine (SNAP) in phenylephrine-precontracted rings without endothelium were attenuated in BDL compared with SO rats. Pretreatment with DMPPO (0.1 microM) enhanced these responses and eliminated the differences between the two groups. Vasodilation to DMPPO itself was also less in BDL rats. The responses to phenylephrine were attenuated in endothelium-rich aorta from BDL relative to SO rats, but they were similar in endothelium-denuded aorta and remained similar despite preincubation with SNAP (0.1 microM) alone or with SNAP and DMPPO. In vivo, BDL rats were vasodilated relative to SO rats; DMPPO (5 mg/kg i.v.) decreased arterial pressure and vascular resistance in both groups equally and caused significant increase in sodium excretion in BDL rats only. In conclusion, the results are in accordance with a possible increase in PDE5 activity in aorta and kidney of cirrhotic rats that results in reduced responses to NO donors and contributes to the increase in sodium retention. PDE5 inhibitors may ameliorate sodium retention in cirrhosis but may worsen vasodilation. Examining the effect of PDE5 inhibitors after chronic administration will be more revealing. PMID- 16373594 TI - ANG II and cardiac myocyte contractility: p38 is not stressed out! PMID- 16373595 TI - Ang-1: Tie-ing up endothelial adhesion? PMID- 16373596 TI - In vivo dispersion in repolarization and arrhythmias in the human heart. PMID- 16373597 TI - Adherent platelets recruit and induce differentiation of murine embryonic endothelial progenitor cells to mature endothelial cells in vitro. AB - The homing and differentiation mechanisms of endothelial progenitor cells (EPCs) at sites of vascular lesions are unclear. To investigate whether platelets play a role in the recruitment and differentiation of EPCs, we made use of a robust mouse embryonic EPC (eEPC) line that reliably differentiates to a mature endothelial phenotype. We found that platelets stimulate chemotaxis and migration of these murine eEPCs. Further, the substantial adhesion of murine eEPCs on immobilized platelets that occurs under dynamic flow conditions is inhibited by neutralizing anti-P-selectin glycoprotein ligand-1 and anti-VLA-4 (beta1 integrin) monoclonal antibodies but not by anti-CD11b (aM-integrin; macrophage antigen-1). Coincubation of murine eEPCs with platelets for 5 days induced differentiation of EPCs to mature endothelial cells as verified by positive von Willebrand factor immunofluorescence and detection of Weibel Palade bodies through electron microscopy. We conclude that platelets may play a critical part in the capture and subsequent differentiation of murine eEPCs at sites of vascular lesions, revealing a possible new role of platelets in neoendothelization after vascular injury. PMID- 16373598 TI - H11 kinase prevents myocardial infarction by preemptive preconditioning of the heart. AB - Ischemic preconditioning confers powerful protection against myocardial infarction through pre-emptive activation of survival signaling pathways, but it remains difficult to apply to patients with ischemic heart disease, and its effects are transient. Promoting a sustained activation of preconditioning mechanisms in vivo would represent a novel approach of cardioprotection. We tested the role of the protein H11 kinase (H11K), which accumulates by 4- to 6 fold in myocardium of patients with chronic ischemic heart disease and in experimental models of ischemia. This increased expression was quantitatively reproduced in cardiac myocytes using a transgenic (TG) mouse model. After 45 minutes of coronary artery occlusion and reperfusion, hearts from TG mice showed an 82+/-5% reduction in infarct size compared with wild-type (WT), which was similar to the 84+/-4% reduction of infarct size observed in WT after a protocol of ischemic preconditioning. Hearts from TG mice showed significant activation of survival kinases participating in preconditioning, including Akt and the 5'AMP activated protein kinase (AMPK). H11K directly binds to both Akt and AMPK and promotes their nuclear translocation and their association in a multiprotein complex, which results in a stimulation of survival mechanisms in cytosol and nucleus, including inhibition of proapoptotic effectors (glycogen synthase kinase 3beta, Bad, and Foxo), activation of antiapoptotic effectors (protein kinase Cepsilon, endothelial and inducible NO synthase isoforms, and heat shock protein 70), increased expression of the hypoxia-inducible factor-1alpha, and genomic switch to glucose utilization. Therefore, activation of survival pathways by H11K preemptively triggers the antiapoptotic and metabolic response to ischemia and is sufficient to confer cardioprotection in vivo equally potent to preconditioning. PMID- 16373599 TI - Impaired angiogenesis in glutathione peroxidase-1-deficient mice is associated with endothelial progenitor cell dysfunction. AB - Several vascular disease are characterized by elevated levels of reactive oxygen species (ROS). Vascular endothelium is protected from oxidant stress by expressing enzymes such as glutathione peroxidase type 1 (GPx-1). In this study, we investigated the effect of vascular oxidant stress on ischemia-induced neovascularization in a murine model of homozygous deficiency of GPx-1. GPx-1 deficient mice showed impaired revascularization following hindlimb ischemic surgery based on laser Doppler measurements of blood flow and capillary density in adductor muscle. GPx-1-deficient mice also showed an impaired ability to increase endothelial progenitor cell (EPC) levels in response to ischemic injury or subcutaneous administration of vascular endothelial growth factor protein. EPCs isolated from GPx-1-deficient mice showed a reduced ability to neutralize oxidative stress in vitro, which was associated with impaired migration toward vascular endothelial growth factor and increased sensitivity to ROS-induced apoptosis. EPCs isolated from GPx-1-deficient mice were impaired in their ability to promote angiogenesis in wild-type mice, whereas wild-type EPCs were effective in stimulating angiogenesis in GPx-1-deficient mice. These data suggest that EPC dysfunction is a mechanism by which elevated levels of ROS can contribute to vascular disease. PMID- 16373600 TI - Increased sarcoplasmic reticulum calcium leak but unaltered contractility by acute CaMKII overexpression in isolated rabbit cardiac myocytes. AB - The predominant cardiac Ca2+/calmodulin-dependent protein kinase (CaMK) is CaMKIIdelta. Here we acutely overexpress CaMKIIdeltaC using adenovirus-mediated gene transfer in adult rabbit ventricular myocytes. This circumvents confounding adaptive effects in CaMKIIdeltaC transgenic mice. CaMKIIdeltaC protein expression and activation state (autophosphorylation) were increased 5- to 6-fold. Basal twitch contraction amplitude and kinetics (1 Hz) were not changed in CaMKIIdeltaC versus LacZ expressing myocytes. However, the contraction-frequency relationship was more negative, frequency-dependent acceleration of relaxation was enhanced (tau(0.5Hz)/tau(3Hz)=2.14+/-0.10 versus 1.87+/-0.10), and peak Ca2+ current (ICa) was increased by 31% (-7.1+/-0.5 versus -5.4+/-0.5 pA/pF, P<0.05). Ca2+ transient amplitude was not significantly reduced (-27%, P=0.22), despite dramatically reduced sarcoplasmic reticulum (SR) Ca2+ content (41%; P<0.05). Thus fractional SR Ca2+ release was increased by 60% (P<0.05). Diastolic SR Ca2+ leak assessed by Ca2+ spark frequency (normalized to SR Ca2+ load) was increased by 88% in CaMKIIdeltaC versus LacZ myocytes (P<0.05; in an multiplicity-of-infection dependent manner), an effect blocked by CaMKII inhibitors KN-93 and autocamtide-2 related inhibitory peptide. This enhanced SR Ca2+ leak may explain reduced SR Ca2+ content, despite measured levels of SR Ca2+-ATPase and Na+/Ca2+ exchange expression and function being unaltered. Ryanodine receptor (RyR) phosphorylation in CaMKIIdeltaC myocytes was increased at both Ser2809 and Ser2815, but FKBP12.6 coimmunoprecipitation with RyR was unaltered. This shows for the first time that acute CaMKIIdeltaC overexpression alters RyR function, leading to enhanced SR Ca2+ leak and reduced SR Ca2+ content but without reducing twitch contraction and Ca2+ transients. We conclude that this is attributable to concomitant enhancement of fractional SR Ca2+ release in CaMKIIdeltaC myocytes (ie, CaMKII-dependent enhancement of RyR Ca2+ sensitivity during diastole and systole) and increased ICa. PMID- 16373601 TI - Differences in vascular bed disease susceptibility reflect differences in gene expression response to atherogenic stimuli. AB - Atherosclerosis occurs predominantly in arteries and only rarely in veins. The goal of this study was to test whether differences in the molecular responses of venous and arterial endothelial cells (ECs) to atherosclerotic stimuli might contribute to vascular bed differences in susceptibility to atherosclerosis. We compared gene expression profiles of primary cultured ECs from human saphenous vein (SVEC) and coronary artery (CAEC) exposed to atherogenic stimuli. In addition to identifying differentially expressed genes, we applied statistical analysis of gene ontology and pathway annotation terms to identify signaling differences related to cell type and stimulus. Differential gene expression of untreated venous and arterial endothelial cells yielded 285 genes more highly expressed in untreated SVEC (P<0.005 and fold change >1.5). These genes represented various atherosclerosis-related pathways including responses to proliferation, oxidoreductase activity, antiinflammatory responses, cell growth, and hemostasis functions. Moreover, stimulation with oxidized LDL induced dramatically greater gene expression responses in CAEC compared with SVEC, relating to adhesion, proliferation, and apoptosis pathways. In contrast, interleukin 1beta and tumor necrosis factor alpha activated similar gene expression responses in both CAEC and SVEC. The differences in functional response and gene expression were further validated by an in vitro proliferation assay and in vivo immunostaining of alphabeta-crystallin protein. Our results strongly suggest that different inherent gene expression programs in arterial versus venous endothelial cells contribute to differences in atherosclerotic disease susceptibility. PMID- 16373602 TI - Enhanced vascular responses to adrenomedullin in mice overexpressing receptor activity-modifying protein 2. AB - Adrenomedullin (AM) levels are elevated in cardiovascular disease, but little is known of the role of specific receptor components. AM acts via the calcitonin receptor-like receptor (CLR) interacting with a receptor-activity-modifying protein (RAMP). The AM1 receptor is composed of CLR and RAMP2, and the calcitonin gene-related peptide (CGRP) receptor of CLR and RAMP1, as determined by molecular and cell-based analysis. This study examines the relevance of RAMP2 in vivo. Transgenic (TG) mice that overexpress RAMP2 in smooth muscle were generated. The role of RAMP2 in the regulation of blood pressure and in vascular function was investigated. Basal blood pressure, acute angiotensin II-raised blood pressure, and cardiovascular properties were similar in wild-type (WT) and TG mice. However, the hypotensive effect of IV AM, unlike CGRP, was enhanced in TG mice (P<0.05), whereas a negative inotropic action was excluded by left-ventricular pressure-volume analysis. In aorta relaxation studies, TG vessels responded in a more sensitive manner to AM (EC50, 8.0+/-1.5 nmol/L) than WT (EC50, 17.9+/-3.6 nmol/L). These responses were attenuated by the AM receptor antagonist, AM(22 52), such that residual responses were identical in all mice. Remaining relaxations were further inhibited by CGRP receptor antagonists, although neither affected AM responses when given alone. Mesenteric and cutaneous resistance vessels were also more sensitive to AM in TG than WT mice. Thus RAMP2 plays a key role in the sensitivity and potency of AM-induced hypotensive responses via the AM1 receptor, providing evidence that this receptor is a selective target for novel therapeutic approaches. PMID- 16373603 TI - Lipoprotein-associated phospholipase A2 and measures of extracoronary atherosclerosis: the Rotterdam Study. AB - OBJECTIVE: Lipoprotein-associated phospholipase A2 (Lp-PLA2) may be a new and independent predictor of cardiovascular events. The effect of Lp-PLA2 may be exerted through effects of the enzyme on the development of atherosclerosis. Therefore, we investigated the association between Lp-PLA2 activity and measures of extracoronary atherosclerosis. METHODS AND RESULTS: Lp-PLA2 activity was determined in a random sample of 1820 participants from the Rotterdam Study, a population-based cohort study in men and women > or =55 years. Common carotid intima-media thickness, carotid plaques, ankle-arm index, and aortic calcification were examined. Atherosclerosis status could be assigned in 1609 participants. The age-adjusted odds ratio of having atherosclerosis at any site for the highest versus the lowest tertile of Lp-PLA2 activity was 1.86 (95% CI, 1.01 to 3.43) in men and 1.60 (95% CI, 1.08 to 2.37) in women. After additional adjustment for cholesterol, these associations attenuated or even disappeared. The odds ratios of having atherosclerosis at specific sites (carotid arteries and aortic-iliac-femoral tract) followed a similar pattern. CONCLUSIONS: Although Lp PLA2 has been found to be independently associated with cardiovascular events, the association with measures of subclinical extracoronary atherosclerosis found in this study strongly attenuated or even disappeared after adjustment for cholesterol. PMID- 16373604 TI - Apolipoprotein E recycling: implications for dyslipidemia and atherosclerosis. AB - After receptor-mediated endocytosis, the intracellular fate of triglyceride-rich lipoproteins (TRLs) is far more complex than the classical degradation pathway of low-density lipoproteins. Once internalized, TRLs disintegrate in peripheral endosomes, followed by a differential sorting of TRL components. Although core lipids and apolipoprotein B are targeted to lysosomes, the majority of TRL derived apolipoprotein E (apoE) remains in peripheral recycling endosomes. This pool of TRL-derived apoE is then mobilized by high-density lipoproteins (HDLs) or HDL-derived apoA-I to be recycled back to the plasma membrane, followed by apoE resecretion and the subsequent formation of apoE-containing HDL. The HDL-induced recycling of apoE is accompanied by cholesterol efflux and involves the internalization and targeting of HDL-derived apoA-I to endosomes containing both apoE and cholesterol. These findings point to a yet unknown intracellular link between TRL-derived apoE, cellular cholesterol transport, and HDL metabolism. Recent studies provide first evidence that impaired recycling of TRL-derived apoE4, but not apoE3, is associated with intracellular cholesterol accumulation, which might explain some well-documented effects of apoE4 on HDL metabolism. This review summarizes the current understanding of apoE recycling and its potential role in the regulation of plasma apoE levels in the postprandial state. PMID- 16373605 TI - Transforming growth factor beta 1 induces neointima formation through plasminogen activator inhibitor-1-dependent pathways. AB - OBJECTIVE: The mechanisms through which transforming growth factor (TGF)-beta1 promotes intimal growth, and the pathways through which TGF-beta1 expression is regulated in the artery wall, are incompletely understood. We used a mouse model to investigate mechanisms of TGF-beta1-induced intimal growth. METHODS AND RESULTS: Adenovirus-mediated overexpression of TGF-beta1 in uninjured carotid arteries of wild-type mice induced formation of a cellular and matrix-rich intima. Intimal growth appeared primarily due to cell migration and matrix accumulation, with only a negligible contribution from cell proliferation. Overexpression of TGF-beta1 also stimulated expression of plasminogen activator inhibitor type 1 (plasminogen activator inhibitor [PAI]-1) in the artery wall. To test the hypothesis that PAI-1 is a critical downstream mediator of TGF-beta1 induced intimal growth, we transduced carotid arteries of PAI-1-deficient (Serpine1(-/-)) mice with the TGF-beta1-expressing vector. Overexpression of TGF beta1 in Serpine1(-/-) arteries did not increase intimal growth, matrix accumulation, cell migration, or proliferation. Moreover, TGF-beta1-transduced arteries of Serpine1(-/-) mice secreted 6- to 10-fold more TGF-beta1 than did arteries of wild-type mice that were infused with the same concentration of the TGF-beta1-expressing vector. CONCLUSIONS: PAI-1 is both a critical mediator of TGF-beta1-induced intimal growth and a key negative regulator of TGF-beta1 expression in the artery wall. PMID- 16373606 TI - Pravastatin inhibits expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) in Watanabe heritable hyperlipidemic rabbits: a new pleiotropic effect of statins. AB - BACKGROUND: LOX-1, a receptor for oxidized low-density lipoprotein (OxLDL), seems to play a critical role in foam cell formation of macrophages (Mphis) and smooth muscle cells (SMC). Inhibition of LOX-1 expression reduces foam cell formation and might influence lipid core formation in atherosclerotic lesions. Because statins are able to downregulate LOX-1 expression in vitro, we examined if pravastatin can be used to reduce LOX-1 expression and lipid core formation in lesions of Watanabe heritable hyperlipidemic (WHHL) rabbits. METHODS AND RESULTS: Pravastatin downregulated LOX-1 expression in cultured human Mphis and in cultured human aortic SMCs. Homozygous WHHL rabbits were treated with 50 mg kg( 1) d(-1) pravastatin for 32 weeks. Immunohistochemical studies revealed that LOX 1 was expressed in intimal Mphis and SMCs of atherosclerotic lesions. The pravastatin-treated rabbits showed, compared with untreated rabbits, a significantly reduced LOX-1 protein and mRNA expression in the aortic arch. Lipid labeling of this aorta region also demonstrated a strong reduction of the ratio of lipid core area/total lesion area in pravastatin-treated rabbits. CONCLUSIONS: The in vivo inhibition of LOX-1 expression by pravastatin demonstrated here represents a new pleiotropic effect of pravastatin. This in vivo inhibition of LOX-1 might be one mechanism for the lipid core reducing effect of pravastatin in atherogenesis. PMID- 16373607 TI - Diet and murine atherosclerosis. AB - Lipid-enriched diets are often used to induce or accelerate the rate of atherosclerotic lesion development in murine models of atherosclerosis. It appears that the induction of persistent hypercholesterolemia to levels > or approximately to 300 mg/dL is required for the development of experimental atherosclerosis in the mouse. A variety of different diets have been used that vary in the level of cholesterol, the level and type of fatty acid, and the absence or presence of cholate. Each of these components as well as the protein source has been shown to influence lipoprotein level and/or atherosclerosis, with dietary cholesterol being the major proatherogenic component. In some instances the effects of these components on the expression of hepatic genes relevant to lipid homeostasis has been observed. An appreciation of the effect of the differences in diet composition on these processes is important to compare results from different atherosclerosis studies, so the composition of the diets used should always be reported or referenced. Cholate should not be used unless its effects are being specifically investigated. PMID- 16373608 TI - Counter-regulatory function of protein tyrosine phosphatase 1B in platelet derived growth factor- or fibroblast growth factor-induced motility and proliferation of cultured smooth muscle cells and in neointima formation. AB - OBJECTIVE: We have previously reported that vascular injury or treatment of cultured vascular smooth muscle cells with platelet-derived growth factor-BB (PDGF-BB) or fibroblast growth factor-2 (FGF2) increases the levels of protein tyrosine phosphatase (PTP)1B. The current study was designed to test the hypothesis that PTP1B attenuates PDGF- or FGF-induced motility and proliferation of cultured cells, as well as neointima formation in injured rat carotid arteries. METHODS AND RESULTS: Treatment of cultured cells with adenovirus expressing PTP1B decreased PDGF-BB- or FGF2-induced cell motility and blocked PDGF-BB- or FGF2-induced proliferation, whereas expression of dominant negative PTP1B (C215S-PTP1B) uncovered the motogenic effect of subthreshold levels of PDGF BB or FGF2, increased neointimal and medial cell proliferation, and induced neointimal enlargement after balloon injury. The inhibitory effect of PTP1B directed against PDGF in cultured cells was associated with dephosphorylation of the PDGFbeta receptor. CONCLUSIONS: PTP1B suppresses cell proliferation and motility in cultured smooth muscle cells treated with PDGF-BB or FGF2, and the phosphatase plays a counter-regulatory role in vascular injury-induced cell proliferation and neointima formation. Taken together with previous studies indicating increased PTP1B levels in cells treated with growth factors, the current findings are the first to report the existence of an inhibitory feedback loop involving PDGF or FGF, and PTP1B in blood vessels. PMID- 16373609 TI - Applications of nanotechnology to atherosclerosis, thrombosis, and vascular biology. AB - The role of nanotechnology in cardiovascular diagnosis is expanding rapidly. The goal of this brief review is to illustrate selected examples of nanosystems that have been applied to the arenas of atherosclerosis, thrombosis, and vascular biology. The technologies for producing targeted nanosystems are multifarious and reflect end uses in many cases. The results to date indicate rapid growth of interest and capability in the field. The future of cardiovascular diagnosis already is being impacted by nanosystems that can both diagnose pathology and treat it with targeted delivery systems. PMID- 16373610 TI - Effect of inhibiting cholesteryl ester transfer protein on the kinetics of high density lipoprotein cholesteryl ester transport in plasma: in vivo studies in rabbits. AB - OBJECTIVE: Inhibitors of cholesteryl ester transfer protein (CETP) have been developed as potential anti-atherogenic agents. Theoretically, however, they may be pro-atherogenic by blocking one of the pathways for removing high-density lipoprotein (HDL) cholesteryl esters (CE) from plasma in the final step of reverse cholesterol transport. Here we describe how CETP inhibition in rabbits impacts on the kinetics of HDL CE transport in plasma. METHODS AND RESULTS: Administration of a CETP inhibitor reduced CETP activity by 80% to 90% and doubled the HDL cholesteryl ester concentration. Multi-compartmental analysis was used to determine HDL CE kinetics in CETP-inhibited and control rabbits after injection of tracer amounts of both native and reconstituted HDL labeled with 3H in the CE moiety. In control rabbits, HDL CE was removed from plasma by both a direct pathway and an indirect pathway after transfer of HDL CE to the very-low density lipoprotein/low-density lipoprotein fraction. In CETP-inhibited rabbits there was an almost complete block in removal via the indirect pathway. This did not compromise the overall removal of HDL CE from plasma, which was not different in control and inhibited animals. CONCLUSIONS: Inhibiting CETP in rabbits does not compromise the removal of HDL CE from plasma. PMID- 16373611 TI - Uremia-specific effects in the arterial media during development of uremic atherosclerosis in apolipoprotein E-deficient mice. AB - OBJECTIVE: Uremia accelerates formation of atherosclerosis-like lesions in apolipoprotein E-deficient (apoE(-/-)) mice. In this study, we compared gene expression patterns in classical and uremic atherosclerosis. METHODS AND RESULTS: High-density oligonucleotide microarray analyses were performed with aortic RNA from 5/6 nephrectomized (NX) and sham-operated mice. After 12 weeks, NX apoE(-/-) mice had more atherosclerosis and 24 genes were differentially expressed as compared with sham apoE(-/-) mice. Nine genes expressed in muscle cells displayed reduced expression (3.3- to 142-fold, P<0.05), whereas osteopontin gene expression was increased 8.7-fold (P<0.05) in NX mice. Studies of NX wild-type mice suggested that the changes in NX apoE(-/-) mice were dependent on hypercholesterolemia. Nevertheless, lesioned versus nonlesioned areas of aortas from nonuremic apoE(-/-) mice with classical atherosclerosis displayed less pronounced reductions in expression of the muscle cell related genes than seen in NX apoE(-/-) mice even though the osteopontin gene expression was increased approximately 15-fold. Electron microscopy showed more vacuolized and necrotic smooth muscle cells within the media underneath both nonlesioned and lesioned intima in NX than in sham apoE(-/-) mice. CONCLUSIONS: The results suggest that uremic vasculopathy in apoE(-/-) mice, in addition to intimal atherosclerosis, is characterized by a uremia-specific medial smooth muscle cell degeneration, which appears to be accentuated by hypercholesterolemia. PMID- 16373612 TI - Atherosclerosis susceptibility loci identified from a strain intercross of apolipoprotein E-deficient mice via a high-density genome scan. AB - OBJECTIVE: Apolipoprotein (apo) E-deficient mice are hypercholesterolemic and develop atherosclerosis on low-fat chow diets; however, the genetic background strain has a large effect on atherosclerosis susceptibility. This study aimed to determine the genetic regions associated with strain effects on lesion area. METHODS AND RESULTS: We performed a strain intercross between atherosclerosis sensitive DBA/2 and atherosclerosis resistant AKR apoE-deficient mice. Aortic root lesion area, total cholesterol, body weights, and complete blood counts were ascertained for 114 male and 95 female F2 progeny. A high-density genome scan was performed using a mouse single nucleotide polymorphism chip yielding 1967 informative polymorphic markers. Quantitative trait locus (QTL) statistical analyses were performed. Novel loci associated with lesion or log lesion area were identified for the female and male F2 cohorts. The atherosclerosis QTLs in female mice reside on chromosomes 15, 5, 3, and 13, and in male mice on chromosomes 17, 18, and 2. QTL were also identified for body weight, total cholesterol, and blood count parameters. CONCLUSIONS: Loci were identified for atherosclerosis susceptibility in a strain intercross study. The identity of the responsible genes at these loci remains to be determined. PMID- 16373613 TI - Identification of the cAMP-responsive enhancer of the murine ABCA1 gene: requirement for CREB1 and STAT3/4 elements. AB - OBJECTIVE: To determine the mechanism by which expression of the murine ABCA1 gene is highly induced by cAMP analogues. METHODS AND RESULTS: ABCA1 mRNA turnover cannot account for its induction by cAMP. Thus cAMP induction of ABCA1 mRNA occurs at a transcriptional level. Shotgun cloning DNA fragments from the murine ABCA1 locus identified a strong cAMP responsive enhancer located in the first intron, which led to 25- to 100-fold cAMP-mediated induction of reporter gene activity. Deletions and mutations of this enhancer led to the identification a cAMP-responsive element (CRE) that was essential for the cAMP induction. Furthermore, the capacity of this CRE site to mediate the cAMP induction required the presence of a STAT3/4 element located 81 bp away. A dominant-negative CREB expression vector inhibited the cAMP induction of ABCA1, demonstrating that CREB was required for cAMP induction of ABCA1 expression in RAW264.7 cells. CONCLUSIONS: Phospho-CREB1 controls the cAMP-mediated induction of murine ABCA1 gene expression through a CRE site acting in cooperation with a nearby STAT element. This CRE site is not conserved in the human ABCA1 gene, explaining why human ABCA1 is not strongly stimulated by cAMP analogs. PMID- 16373614 TI - OxLDL-IgG immune complexes induce survival of human monocytes. AB - OBJECTIVE: Immune complexes containing oxidatively modified low-density lipoprotein (oxLDL) particles are deposited in human atherosclerotic lesions during atherogenesis. Here we studied whether OxLDL-IgG immune complexes (OxLDL IgG ICs) affect survival of human monocytes. METHODS AND RESULTS: As demonstrated by light microscopy, and analysis of cell proliferation, caspase-3 activity, and DNA fragmentation, OxLDL-IgG ICs promoted survival of cultured human monocytes by decreasing their spontaneous apoptosis. OxLDL-IgG ICs induced a concentration dependent production of the major monocyte growth factor, monocyte colony stimulating factor (M-CSF), by the monocytes, but its inhibition was without effect on OxLDL-IgG IC-induced monocyte survival. Rather, OxLDL-IgG ICs induced rapid phosphorylation of Akt, suggesting a direct anti-apoptotic effect mediated by cross-linking of Fcgamma receptors. Experiments with receptor blocking antibodies revealed that the OxLDL-IgG IC-induced monocyte survival was mediated by Fcgamma receptor I. CONCLUSIONS: The results show that OxLDL-IgG ICs promote survival of monocytes by cross-linking Fcgamma receptor I and activating Akt dependent survival signaling. The results reveal a novel mechanism by which an immune reaction toward oxLDL can play a role in the accumulation of macrophages in human atherosclerotic lesions. PMID- 16373615 TI - Stereospecific and redox-sensitive increase in monocyte adhesion to endothelial cells by homocysteine. AB - OBJECTIVE: Previous studies have shown that elevated homocysteine (Hcy) levels promote the development of atherosclerotic lesions in atherosclerosis-prone animal models. There is evidence that oxidant stress contributes to Hcy's deleterious effects on the vasculature. The accumulation and adhesion of monocytes to the vascular endothelium is a critical event in the development of atherosclerosis. We investigated the effects of Hcy on the interaction between human endothelial cells (EC) (EC line EA.hy 926 and primary human umbilical vein EC [HUVEC]) and the monocytic cell line THP-1, and the impact of vascular oxidant stress and redox-sensitive signaling pathways on these events. METHODS AND RESULTS: L-Hcy, but not D-Hcy, increases the production of reactive oxygen species inside EC, enhances nuclear factor(NF)-kappaB activation, and stimulates intercellular adhesion molecule-1 (ICAM-1) RNA transcription and cell surface expression. This leads to a time- and dose-dependent increase in monocyte adhesion to ECs. Pretreatment of ECs with superoxide scavengers (MnTBAP and Tiron) or with an inhibitor of NF-kappaB activation abolished Hcy-induced monocyte adhesion, ICAM-1 expression, and nuclear translocation of NF-kappaB. CONCLUSIONS: These findings suggest that reactive oxygen species produced under hyperhomocysteinemic conditions may induce a proinflammatory situation in the vessel wall that initiates and promotes atherosclerotic lesion development. PMID- 16373616 TI - Serum lipoprotein lipase concentration and risk for future coronary artery disease: the EPIC-Norfolk prospective population study. AB - BACKGROUND: Lipoprotein lipase (LPL) is associated with coronary artery disease (CAD) risk, but prospective population data are lacking. This is mainly because of the need for cumbersome heparin injections, which are necessary for LPL measurements. Recent retrospective studies, however, indicate that LPL concentration can be reliably measured in serum that enabled evaluation of the prospective association between LPL and future CAD. METHODS AND RESULTS: LPL concentration was determined in serum samples of men and women in the EPIC Norfolk population cohort who developed fatal or nonfatal CAD during 7 years of follow-up. For each case (n=1006), 2 controls, matched for age, sex, and enrollment time, were identified. Serum LPL concentration was lower in cases compared with controls (median and interquartile range: 61 [43-85] versus 66 [46 92] ng/mL; P<0.0001). Those in the highest LPL concentration quartile had a 34% lower risk for future CAD compared with those in the lowest quartile (odds ratio [OR] 0.66; confidence interval [CI], 0.53 to 0.83; P<0.0001). This effect remained significant after adjustment for blood pressure, diabetes, smoking, body mass index, and low-density lipoprotein (LDL) cholesterol (OR, 0.77; CI, 0.60 0.99; P=0.02). As expected from LPL biology, additional adjustments for either high-density lipoprotein cholesterol (HDL-C) or triglyceride (TG) levels rendered loss of statistical significance. Of interest, serum LPL concentration was positively linear correlated with HDL and LDL size. CONCLUSIONS: Reduced levels of serum LPL are associated with an increased risk for future CAD. The data suggest that high LPL concentrations may be atheroprotective through decreasing TG levels and increasing HDL-C levels. PMID- 16373617 TI - High plasma levels of osteopontin in patients with restenosis after percutaneous coronary intervention. PMID- 16373620 TI - Lipoprotein-associated phospholipase A2 and cardiovascular risk: state of the evidence and future directions. PMID- 16373621 TI - HIV protease inhibitors and hyperlipidemia: a fatty acid connection. PMID- 16373622 TI - Tightening the barrier: mechanical forces and the control of endothelial permeability. PMID- 16373623 TI - Impaired endothelium-dependent and -independent vasodilation in young female athletes with exercise-associated amenorrhea. PMID- 16373624 TI - Aldosterone promotes thrombosis formation after arterial injury in mice. PMID- 16373625 TI - Multilevel educational program for emergency medical services. PMID- 16373626 TI - Is there an interaction between pravastatin and clinical events other than vasospasm in patients with aneurysmal subarachnoid hemorrhage? PMID- 16373627 TI - Embolus detection and differentiation using multifrequency transcranial Doppler. PMID- 16373628 TI - Gender and carotid artery stenosis. PMID- 16373629 TI - The call for multicenter studies of pediatric stroke. PMID- 16373630 TI - Shunt-associated migraine responds favorably to atrial septal repair: a case control study. AB - BACKGROUND AND PURPOSE: Transcatheter closure of patent foramen ovale (PFO) has been reported to improve migraine in patients with cerebrovascular disorders in noncontrolled studies. The aim of the study was to compare the course of migraine assessed prospectively over a 12-month period in symptomatic (for cerebrovascular disease) and asymptomatic patients undergoing PFO closure and in patients with PFO treated medically. METHODS: Twenty-three stroke symptomatic (SS; 39+/-10 years of age; males/females [M/F] 5/18) and 27 stroke asymptomatic (SA; 40+/-12 years of age; M/F 5/22) patients with migraine underwent PFO closure. Twenty seven patients with migraine and PFO (controls [CTRLS]; 36+/-11 years of age; M/F 4/23) were followed up medically. Migraine severity was assessed at baseline with a scale that takes into account the frequency, duration, and intensity of the attacks and the occurrence of aura (score range 0 to 10). Six months later, the patients were given a structured diary to annotate monthly with the same scale the characteristics of the attacks for the next 6 months. By the end of 1 year, the migraine score was averaged for the last 6 months. RESULTS: Baseline severity of migraine did not differ between groups (6.3 to 6.1 and 6.7 in SS, SA, and CTRLS groups, respectively). At the 1-year assessment, the overall migraine score had significantly improved by 3.7 and 2.8 points in SS and SA, respectively (P<0.001 on repeated-measure ANOVA), whereas it had nonsignificantly worsened by 0.1 points in CTRLS. Multiple linear regression analysis showed that the improvement in SS and SA was independent of migraine type, age, and cerebrovascular risk factors. Twenty-one of 21 patients with migraine with aura in the CTRLS group still had aura at the end of follow-up, whereas only 3 of 14 among SA and 4 of 19 among SS continued to have migraine preceded by aura (P<0.0001 on Fisher exact test). CONCLUSIONS: Compared with medical treatment, closure of PFO brings about a significant overall improvement in migraine. This seems to occur irrespective of migraine type and of previous cerebrovascular disease. In addition to the overall improvement, in migraine with aura, the occurrence of aura is dramatically reduced. PMID- 16373631 TI - Metabolic syndrome as an independent risk factor of silent brain infarction in healthy people. AB - BACKGROUND AND PURPOSE: Metabolic syndrome (MetS) is associated with an increased risk of the subsequent development of cardiovascular disease or stroke. Moreover, a silent brain infarction (SBI) can predict clinical overt stroke or dementia. We examined the associations between SBI and MetS in apparently healthy individuals. METHODS: We evaluated 1588 neurologically healthy subjects (927 males and 661 females) who underwent brain MRI at Seoul National University Hospital Healthcare System Gangnam Center. MetS was defined using the criteria of the National Cholesterol Education Program Adult Treatment Panel III. We examined associations between full syndrome (> or =3 of the 5 conditions) as well as its components and SBI by controlling possible confounders. RESULTS: Eighty-eight (5.5%) were found to have > or =1 SBI on MRI. Age was found to be significantly related to SBI prevalence (odds ratio [OR], 1.06; 95% CI, 1.04 to 1.09). A history of coronary artery disease was associated with an elevated odds ratio of SBI (OR, 2.83; 95% CI, 1.38 to 5.82), and MetS was significantly associated with SBI (OR, 2.18; 95% CI, 1.38 to 3.44). The components model of MetS showed a strong significance between an elevated blood pressure (OR, 3.75; 95% CI, 2.05 to 6.85) and an impaired fasting glucose (OR, 1.74; 95% CI, 1.08 to 2.80) and the risk of SBI. CONCLUSIONS: MetS was found to be significantly associated with SBI. This finding has clinical utility in terms of identifying healthy people at increased risk of developing SBI. PMID- 16373632 TI - Microbubble administration accelerates clot lysis during continuous 2-MHz ultrasound monitoring in stroke patients treated with intravenous tissue plasminogen activator. AB - BACKGROUND AND PURPOSE: We sought to evaluate the effects of administration of microbubbles (MBs) on the beginning, speed, and degree of middle cerebral artery (MCA) recanalization during systemic thrombolysis and continuous 2-MHz pulsed wave transcranial Doppler (TCD) monitoring. METHODS: We evaluated 111 patients with acute stroke attributable to MCA occlusion treated with intravenous tissue plasminogen activator (tPA). Thirty-eight patients were treated with tPA plus continuous 2-hour TCD monitoring plus 3 doses of 2.5 g (400 mg/mL) of galactose based MBs given at 2, 20, and 40 minutes after tPA bolus (MB group). These patients were compared with 73 patients who were allocated to receive tPA plus continuous 2-hour TCD ultrasound (US) monitoring (tPA/US group) or tPA plus placebo monitoring (tPA group), most of whom were enrolled in a previous study of US-enhanced thrombolysis. The beginning, degree, and time to maximum completeness of recanalization during the first 2 hours of tPA bolus were recorded. RESULTS: Median prebolus National Institutes of Health Stroke Scale (NIHSS) score was 18. Eighty patients (72%) had a proximal and 31 (28%) a distal MCA occlusion on TCD. Thirty-seven patients (33%) received tPA/US, 38 (34%) received tPA/US/MB, and 36 (32%) were treated with tPA alone. Stroke severity, time to treatment, location of MCA occlusion, and presence of carotid artery disease were similar among groups. Two-hour recanalization was seen in 14 (39%), 25 (68%), and 27 patients (71%) in the tPA, tPA/US, and tPA/US/MB groups, respectively (P=0.004). Two-hour complete recanalization rate was significantly (P=0.038) higher in the tPA/US/MB group (54.5%) compared with tPA/US (40.8%) and tPA (23.9%) groups. The time to beginning of recanalization after tPA bolus was 26+/-18 minutes in the tPA/US group and 19+/-12 minutes in the tPA/US/MB group (P=0.12). Four patients (3.6%) experienced symptomatic intracranial hemorrhage: 2 (5.5%), 1 (2.7%), and 1 patient (2.6%) who received tPA only, tPA/US, and tPA/US/MB, respectively, experienced symptomatic intracranial hemorrhage. At 24 hours, 31%, 41%, and 55% of tPA, tPA/US, and tPA/US/MB improved >4 points in the NIHSS score. CONCLUSIONS: Administration of MBs induces further acceleration of US-enhanced thrombolysis in acute stroke, leading to a more complete recanalization and to a trend toward better short- and long-term outcome. PMID- 16373633 TI - Stroke and encephalopathy after cardiac surgery: the search for the holy grail. PMID- 16373634 TI - Combined effects of hemoglobin A1c and C-reactive protein on the progression of subclinical carotid atherosclerosis: the INVADE study. AB - BACKGROUND AND PURPOSE: Glycohemoglobin (hemoglobin A1c [HbA1c]) and high sensitivity C-reactive protein (hsCRP) are risk indicators for atherosclerosis. Limited information exists regarding the combined effects of inflammation and hyperglycemia. We investigated the joint effects of both parameters on early carotid atherosclerosis progression and major vascular events in diabetic and nondiabetic subjects. METHODS: We analyzed the data of INVADE (Intervention Project on Cerebrovascular Diseases and Dementia in the Community of Ebersberg, Bavaria), a prospective, population-based study conducted in 3534 subjects (mean age, 69 years). In addition to common risk factors, measurements of carotid intima-media thickness (IMT), hsCRP, and HbA1c were performed at baseline and after 2 years of follow-up. RESULTS: For the entire population, IMT progression was significantly related to HbA1c (P=0.003) but not to hsCRP (P=0.06) after risk factor adjustment. The interaction hsCRPxHbA1c was highly significant (P=0.001), and the most pronounced IMT progression was seen in subjects with both parameters in the fourth quartiles compared with subjects with both parameters in the first quartiles (0.028 [0.025, 0.031] versus 0.012 mm/year [0.007, 0.019]; P=0.0013). We observed a significant joint effect of HbA1c and hsCRP on IMT progression in the diabetic (n=882) as well as the nondiabetic subgroup (n=2652). Subjects with HbA1c and hsCRP in the upper 2 quartiles had an increased risk for new vascular events (adjusted hazard ratio in diabetics: 4.3 [1.8, 7.3]; P=0.001; nondiabetics: 2.9 [1.6, 4.7]; P=0.001). CONCLUSIONS: The combination of hyperglycemia and inflammation is associated with an advanced early carotid atherosclerosis progression and an increased risk of new vascular events in diabetic as well as nondiabetic subjects. PMID- 16373635 TI - Role of NAD(P)H oxidase in alcohol-induced impairment of endothelial nitric oxide synthase-dependent dilation of cerebral arterioles. AB - BACKGROUND AND PURPOSE: Our goal was to determine whether NAD(P)H oxidase is involved in impaired endothelial nitric oxide synthase (eNOS)-dependent reactivity of cerebral arterioles during chronic alcohol consumption. METHODS: Sprague-Dawley rats were fed with an alcohol diet for 2 to 3 months. We determined the effects of acute and chronic treatment with an NAD(P)H oxidase inhibitor, apocynin, on responses of pial arterioles to eNOS-dependent agonists (acetylcholine and ADP) and an eNOS-independent agonist (nitroglycerin). Expression of NAD(P)H oxidase in pial arterioles was measured with the use of real-time polymerase chain reaction and Western blot analysis, and superoxide production was measured with the use of lucigenin-enhanced chemiluminescence. RESULTS: Vasodilation in response to acetylcholine and ADP, but not nitroglycerin, was significantly less in alcohol-fed rats. Treatment with apocynin did not alter vasodilation in non-alcohol-fed rats but significantly improved impaired vasodilation in alcohol-fed rats. In addition, an upregulation of p47phox in pial arterioles was found in alcohol-fed rats. Furthermore, alcohol consumption increased superoxide production under basal conditions and in the presence of ADP and NAD(P)H. CONCLUSIONS: Our findings suggest that NAD(P)H oxidase plays a role in chronic alcohol consumption-induced impairment of eNOS dependent dilation of cerebral arterioles. PMID- 16373636 TI - Stroke and encephalopathy after cardiac surgery: an update. AB - BACKGROUND AND PURPOSE: As a result of advances in surgical, anesthetic, and medical management, cardiac surgery can now be performed on older, sicker patients, some of whom have had prior cardiac interventions. As surgical mortality has declined in recent years, attention has focused on the complications of stroke and encephalopathy after cardiac surgery. SUMMARY OF REVIEW: Patients with preexisting cerebrovascular disease are at increased risk for these untoward neurological outcomes, which are associated with longer lengths of hospital stay, higher costs, and greater mortality. The mechanisms underlying these neurological events may include microemboli and hypoperfusion during surgery, and postoperative atrial fibrillation. Predictive models, based on information available before surgery, allow identification of these "high risk" patients. CONCLUSIONS: Establishing the degree of functionally significant vascular disease of the brain before surgery should be an essential part of the preoperative evaluation, particularly when modifications in surgical technique or novel neuroprotective agents are being evaluated. PMID- 16373637 TI - Quantitative assessment of cerebral hemodynamics using perfusion-weighted MRI in patients with major cerebral artery occlusive disease: comparison with positron emission tomography. AB - BACKGROUND AND PURPOSE: Cerebrovascular reactivity (CVR) to acetazolamide is a key parameter in determining the severity of hemodynamic impairment in patients with major cerebral artery occlusive disease. Perfusion-weighted MRI (PW MRI) can measure the cerebral blood volume (CBV) as an indicator of hemodynamic impairment. CBV measured by PW MRI was compared with CVR measured by positron emission tomography (PET). METHODS: Twelve normal subjects and 17 patients with major cerebral artery occlusive disease underwent PW MRI and PET. The images were coregistered with 3-dimensional spoiled gradient-recalled acquisition images. Quantitative PW MRI-CBV maps were generated using the indicator dilution method with arterial input function. One large cortical region of interest for each unilateral middle cerebral artery territory was determined on each image. PET-CVR was calculated by measuring cerebral blood flow before and after acetazolamide challenge. RESULTS: A significant negative correlation was observed between PW MRI-CBV and PET-CVR (r=-0.713; P<0.0001). PW MRI-CBV higher than the mean +2 SD obtained in normal subjects (15.2 mL/100 g) was defined as elevated and PET-CVR lower than the mean -2 SD obtained in normal subjects (15.1%) was defined as reduced. Assuming the PET-CVR as the true determinant of hemodynamic impairment, PW MRI-CBV provided 80.0% sensitivity and 91.7% specificity, with 80.0% positive predictive value for detecting patients with reduced CVR. CONCLUSIONS: The PW MRI CBV method can simply and accurately identify patients with hemodynamic impairment without exposure to ionizing radiation. PMID- 16373638 TI - Ineffectiveness of prism adaptation on spatial neglect signs. AB - BACKGROUND AND PURPOSE: To evaluate the effect of prism adaptation (PA) on spatial neglect signs. METHODS: Ten patients (hemianopia in 6) and 8 control subjects were included. Tasks were reading single words, nonwords, and a text, bell cancellation, line bisection, and scene drawing, before and after adaptation, with either deviating or neutral prisms. RESULTS: Errors were more frequent in patients but without any specific effect of PA. We found partial improvement at the late sessions, independent of prisms. CONCLUSIONS: We did not confirm the efficacy of PA on spatial neglect. PMID- 16373639 TI - Is there any benefit from staged carotid and coronary revascularization using carotid stents? A single-center experience highlights the need for a randomized controlled trial. AB - BACKGROUND AND PURPOSE: To assess the benefits of carotid artery stenting before coronary artery bypass surgery to reduce the risk of stroke occurring during the cardiac procedure. METHODS: A prospective cohort study was performed in patients undergoing carotid artery stenting before coronary artery bypass surgery, or combined bypass and valve replacement procedures, to assess the procedures effectiveness in stroke prevention. Outcome measures including 30-day post stenting and cardiac surgery neurological complication and all-cause mortality rates were assessed. RESULTS: A total of 52 patients were included. Two patients underwent aortic valve replacements at the same time as coronary revascularization. No neurological complications occurred because of the stenting procedure. One cardiac death not related to coronary artery bypass surgery occurred in the 30-day follow-up period for the stent procedure. An additional 6 (11.5%) outcome events (3 strokes and 3 deaths) occurred in the 30-day follow-up period after the cardiac procedure. Three patients died of cardiac causes while awaiting their cardiac bypass procedure. CONCLUSIONS: Our results are comparable to those in patients that undergo staged or combined carotid endarterectomy before cardiac surgery. Our small cohort study adds to the limited world literature on the subject but is not sufficiently powered to recommend alterations in practice. PMID- 16373640 TI - Eliciting information on differential sensation of heat in those with and without poststroke aphasia using a visual analogue scale. AB - BACKGROUND AND PURPOSE: Aphasia can result in an inability to communicate the presence, location, or intensity of pain. Although visual analogue scales (VASs) exist, it is unknown whether they are useful in assessing pain in individuals with aphasia. The objective was to determine whether those with poststroke aphasia could respond differentially to thermal stimuli of varying intensities using a standardized VAS. METHODS: Five groups of participants were assessed: those without stroke, those with stroke but without aphasia, and 3 groups with varying degrees of aphasia. A 10-cm vertical VAS was used to measure responses to varying thermal intensities delivered on the participant's forearm. RESULTS: Across all 5 groups, a similar proportion demonstrated ability to discriminate between 2 temperatures (chi2=1.899; P=0.75). When presented with 4 temperatures, all groups performed more poorly, yet with similar success rates across groups (chi2=0.1267; P=0.88). The repeated-measures ANOVA revealed no effect of group but a significant effect of temperature (P<0.0001). CONCLUSIONS: A VAS may be useful in clinical identification of differing intensities of stimuli in a substantial proportion of those with aphasia. PMID- 16373641 TI - The significance of various blood pressure indices for long-term stroke, coronary heart disease, and all-cause mortality in men: the Israeli Ischemic Heart Disease study. AB - BACKGROUND AND PURPOSE: Studies of blood pressure (BP) indices as disease predictors have offered conflicting conclusions. We compare pulse pressure (PP), systolic BP (SBP) and diastolic BP (DBP), and mean arterial pressure (MAP) as risk markers for long-term mortality with emphasis on stroke. METHODS: Male civil servants (40 to 65 years of age; n=9611) were examined in 1963 and followed up until 1986. Multivariate analysis was used to assess the association between BP indices and subsequent mortality. Stroke mortality was analyzed separately for initially normotensive (SBP< or =140 and DBP< or =90 mm Hg), hypertensive (SBP>140 and DBP>90 mm Hg), and men with isolated systolic hypertension (ISH; SBP>140 and DBP< or =90 mm Hg). RESULTS: During follow-up, 3167 men died, including 932 of coronary heart disease and 339 of stroke. All 4 BP indices were predictive of fatal stroke among hypertensive and normotensive men (hazard ratios [HRs] fluctuated between 1.59 and 2.51). In models with 2 BP indices among normotensive men, SBP but not DBP remained a predictor of stroke mortality. MAP and PP were independent predictors of stroke mortality. Among hypertensive men, SBP and DBP were independent predictors of stroke mortality (HRs, 1.68 and 1.51, respectively). MAP but not PP remained a predictor of stroke mortality. In men with ISH, the 4 BP indices predicted fatal stroke, with HRs fluctuating between 1.24 and 2.04. CONCLUSIONS: All 4 BP indices were predictors of stroke mortality among hypertensive and normotensive men, with DBP possibly the weaker predictor among the latter. Models with 2 BP indices yielded complex associations. PMID- 16373642 TI - The impact of functional status at three months on long-term survival after spontaneous intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Few studies have assessed long-term prognosis and risk factors for death after spontaneous intracerebral hemorrhage (ICH). Patients who survive the acute phase may run different prognoses, depending on their disability, treatment, and lifestyle. The present study was performed to find out the predictors for long-term mortality after ICH. METHODS: We assessed 7-year prognosis in a population-based cohort of patients who had survived the first 3 months after ICH (n=140). Controls (n=206) living in the same geographical area were randomly drawn from the population register and followed up for the same time. RESULTS: Seven-year mortality was significantly higher in ICH patients than in controls (32.9 and 19.4%, respectively; P=0.0034). The annual risk for death in ICH patients was 5.6%, and the annual risk for fatal recurrent ICH was 1.3%. The ICH patients with good recovery at 3 months showed similar risk for death as controls. Recurrent ICH and pneumonia were the most common causes of death in ICH patients. Cigarette smoking, age, and diabetes seemed to increase the risk for death in patients and controls. CONCLUSIONS: Survivors of ICH run a higher long term risk for death than age- and sex-matched controls. However, those who show good recovery at 3 months run a similar outcome as controls. PMID- 16373643 TI - Xenon provides short-term neuroprotection in neonatal rats when administered after hypoxia-ischemia. AB - BACKGROUND AND PURPOSE: Brain injury after hypoxic-ischemic insults evolves via an apoptotic/necrotic cascade. Glutamate over release and N-methyl-d-aspartate (NMDA) receptor over activation (excitotoxicity) are believed to trigger this process. Xenon is a nontoxic anesthetic gas that reduces neurotransmitter release and functionally antagonizes NMDA receptors. Administering xenon to hypoxic ischemic newborns might be clinically effective if the neurotoxic processes continue evolving after delivery. We sought to determine whether xenon administration after the initial hypoxic-ischemic insult was neuroprotective. METHODS: Fifty 7-day-old rats received a 90-minute hypoxic insult after unilateral carotid ligation. They were then randomized to breathe 1 of 2 gas mixtures for 3 hours: 50% Xe/30% O2/20% N2 or 30% O2/70% N2. RESULTS: One week after hypoxic-ischemic survival, significant global protection was seen in the xenon group (80% less injury); cortex/white matter (88% versus 25%), hippocampus (62% versus 0%), basal ganglia (81% versus 25%), and thalamus (50% versus 0%; percentage of global damage score in nonxenon versus xenon groups, respectively). CONCLUSIONS: Three hours of xenon administration commenced after hypoxia-ischemia in neonatal rats provides short-term neuroprotection. This finding suggests that treatment with xenon after perinatal asphyxia would also be neuroprotective. Because xenon does not cause other neurotoxic effects and has demonstrated minimal side effects in extensive anesthesia studies, it would make an ideal candidate for the treatment after human perinatal hypoxia-ischemia. PMID- 16373644 TI - Association of Phosphodiesterase 4D with ischemic stroke: a population-based case control study. AB - BACKGROUND AND PURPOSE: The Phosphodiesterase 4D (PDE4D) gene was reported recently to be associated with ischemic stroke in an Icelandic population. The association was found predominately with large vessel and cardioembolic stroke. However, 2 recent reports were unable to confirm this association, although a trend toward association with cardioembolic stroke was reported. None of the reports included significant proportions of blacks. We tested for genotype and haplotype association of polymorphisms of the PDE4D gene with ischemic stroke in a population-based, biracial, case-control study. METHODS: A total of 357 cases of ischemic stroke and 482 stroke-free controls from the same community were examined. Single nucleotide polymorphisms (SNPs) were chosen based on significant associations reported previously. Linkage disequilibrium (LD), SNP, and haplotype association analysis was performed using PHASE 2.0 and Haploview 3.2. RESULTS: Although several univariate associations were identified, only 1 SNP (rs2910829) was found to be significantly associated with cardioembolic stroke among both whites and blacks. The rs152312 SNP was associated with cardioembolic stroke among whites after multiple comparison corrections. The same SNP was not associated with cardioembolic stroke among blacks. However, significant haplotype association was identified for both whites and blacks for all ischemic stroke, cardioembolic stroke, and stroke of unknown origin. Haplotype association was identified for small vessel stroke among whites. CONCLUSIONS: PDE4D is a risk factor for ischemic stroke and, in particular, for cardioembolic stroke, among whites and blacks. Further study of this gene is warranted. PMID- 16373645 TI - CCM2 expression parallels that of CCM1. AB - BACKGROUND AND PURPOSE: Mutations in CCM2 (MGC4607 or malcavernin) cause familial cerebral cavernous malformation (CCM), an autosomal dominant neurovascular disease. Both the function of this molecule and the pathogenesis of the disease remain elusive. METHODS: We analyzed the mRNA expression of Ccm1 and Ccm2 in the embryonic and postnatal mouse brain by in situ hybridization. Subsequently, we generated CCM2-specific polyclonal antibodies and tested their specificity using transient transfection experiments in various cell lines. We then investigated CCM2 protein expression in cerebral and extracerebral tissues by Western blot analysis as well as immunohistochemistry and compared these results with CCM1 (KRIT1) protein expression. RESULTS: In situ analysis shows similar temporal and spatial expression patterns for Ccm1 and Ccm2, although Ccm1 expression appears more widespread. Immunohistochemical analysis shows that CCM2 is expressed in various human organs, most noticeably in the arterial vascular endothelium. As is the case with CCM1, CCM2 is not expressed in other vascular wall elements such as smooth muscle cells or the venous circulation. Within cerebral tissue, it is also expressed in pyramidal neurons, astrocytes, and their foot processes. In extracerebral tissues, CCM2 is present in various epithelial cells necessary for blood-organ barrier formation. CONCLUSIONS: CCM1 and CCM2 have similar expression patterns during development and postnatally thereafter. Given the fact that the disease phenotypes caused by mutations in either gene are clinically and pathologically indistinguishable, the significant overlap in expression pattern supports the hypothesis that both molecules are involved in the same pathway important for central nervous system vascular development. PMID- 16373646 TI - Prognostic value of cerebral perfusion-computed tomography in the acute stage after subarachnoid hemorrhage for the development of delayed cerebral ischemia. AB - BACKGROUND AND PURPOSE: Delayed cerebral ischemia (DCI) is an important cause of death and disability after subarachnoid hemorrhage. We studied the additional prognostic value of brain perfusion to 3 established predictors (age, clinical condition on admission, and amount of subarachnoid blood) for the development of DCI. METHODS: We included 69 patients scanned with perfusion--computed tomography within 72 hours after subarachnoid hemorrhage. For each patient, we determined cerebral blood flow (CBF) ratios of prespecified opposite regions of interest and the 3 established predictors. We calculated adjusted hazard ratios (HRs) for the CBF ratio and the 3 established predictors by means of multivariate analysis (Cox regression). The additional prognostic value of CBF ratios was assessed by comparing the area under the receiver operating characteristic curve (AUC) of 2 models: 1 with and 1 without addition of the CBF ratio to the 3 established predictors. RESULTS: The CBF ratio was an independent predictor for the development of DCI (HR, 0.63; 95% CI, 0.46 to 0.86) as was clinical condition (HR, 1.47; 95% CI, 1.01 to 2.13). By adding the CBF ratio to the model with the 3 established predictors, the AUC of the receiver operating characteristic curve increased from 0.76 (95% CI, 0.65 to 0.89) to 0.81 (95% CI, 0.71 to 0.91). This trend toward an increased AUC suggests an improved predictive value. CONCLUSIONS: The CBF ratio is an independent predictor for the development of DCI and can contribute to a better identification of patients at high risk for DCI. PMID- 16373647 TI - Comprehensive transcriptome of proteases and protease inhibitors in vascular cells. AB - BACKGROUND AND PURPOSE: Smooth muscle cells, endothelial cells, and macrophages are essential components of the vasculature, of which the homeostatic gene expression participate importantly in the maintenance of vascular wall integrity. The pathogenesis of vascular diseases, such as cerebral ischemia, atherosclerosis, and abdominal aortic aneurysms, often associates with inflammation and altered gene expression, including proteolytic enzymes that play multiple and important roles in extracellular matrix degradation, cell proliferation and migration, and latent enzyme or growth factor activation. METHODS: Human saphenous vein smooth muscle cells, endothelial cells, and monocyte-derived macrophages from 3 independent donors were stimulated with interleukin 1beta, interferon gamma, tumor necrosis factor alpha, basic fibroblast growth factor, and vascular endothelial growth factor, 5 common proinflammatory mediators often found in diseased human microvessels and macrovessels. Quantitative real-time PCR was used to examine the mRNA levels of 49 proteolytic enzymes and their inhibitors, selected from 4 protease families, in these vascular cells. CONCLUSIONS: Although primary cultured cells from different donors may behave differently in response to these proinflammatory cytokines, data from this study revealed a broad view of vascular cell protease expression profiles under inflammatory conditions, critical to studies of inflammation-associated vascular tissue remodeling. PMID- 16373648 TI - Efficacy of B vitamins in lowering homocysteine in older men: maximal effects for those with B12 deficiency and hyperhomocysteinemia. AB - BACKGROUND AND PURPOSE: A higher plasma concentration of total homocysteine (tHcy) is associated with a greater risk of cardiovascular events. Previous studies, largely in younger individuals, have shown that B vitamins lowered tHcy by substantial amounts and that this effect is greater in people with higher tHcy and lower folate levels. METHODS: We undertook a 2-year, double-blind, placebo controlled, randomized trial in 299 men aged > or =75 years, comparing treatment with a daily tablet containing 2 mg of folate, 25 mg of B6, and 400 microg of B12 or placebo. The study groups were balanced regarding age (mean+/-SD, 78.9+/-2.8 years), B vitamins, and tHcy at baseline. RESULTS: Among the 13% with B12 deficiency, the difference in mean changes in treatment and control groups for tHcy was 6.74 micromol/L (95% CI, 3.94 to 9.55 micromol/L) compared with 2.88 micromol/L (95% CI, 0.07 to 5.69 micromol/L) for all others. Among the 20% with hyperhomocysteinaemia, the difference between mean changes in treatment and control groups for men with high plasma tHcy compared with the rest of the group was 2.8 micromol/L (95% CI, 0.6 to 4.9 micromol/L). Baseline vitamin B12, serum folate, and tHcy were significantly associated with changes in plasma tHcy at follow-up (r=0.252, r=0.522, and r=-0.903, respectively; P=0.003, <0.001, and <0.001, respectively) in the vitamin group. CONCLUSIONS: The tHcy-lowering effect of B vitamins was maximal in those who had low B12 or high tHcy levels. Community dwelling older men, who are likely to be deficient in B12 or have hyperhomocysteinemia, may be most likely to benefit from treatment with B vitamins. PMID- 16373649 TI - Markers of inflammation and infection influence the outcome of patients with baseline asymptomatic carotid lesions: a 5-year follow-up study. AB - BACKGROUND AND PURPOSE: It is still in debate whether the evaluation of markers of infection and inflammation may be of importance for cerebrovascular and cardiovascular prevention, and we aimed to investigate this field in a prospective 5-year clinical follow-up study in patients with early stages of atherosclerosis. METHODS: We studied 668 subjects divided in 3 groups according to the results of carotid ultrasound examination: (1) normal subjects, if intima media thickness (IMT) was <0.9 mm; (2) with IMT, if IMT was between 0.9 and 1.5 mm; and (3) with asymptomatic carotid plaque, if IMT was >1.5 mm. Traditional cardiovascular risk factors were investigated, and laboratory analysis included measurement of plasma lipids, fibrinogen, C-reactive protein, IgG antibodies for helicobacter pylori (HP), cytotoxic HP, cytomegalovirus, and chlamydia pneumoniae. RESULTS: Cerebrovascular or cardiovascular events were registered in 18% of patients during the follow-up, and at multivariate analysis we found that the high levels of fibrinogen (P<0.0001) and C-reactive protein (P=0.014), the seropositivity to cytotoxic HP (P=0.001) and chlamydia pneumoniae (P=0.026), the presence of IMT or asymptomatic carotid plaque (P<0.0001), and the total burden of infections (P<0.0001) were the variables predictive of the clinical events. CONCLUSIONS: Beyond traditional cardiovascular risk factors, markers of inflammation and infections seem to significantly influence the occurrence of cerebrovascular and cardiovascular events in patients with baseline asymptomatic carotid lesions. PMID- 16373650 TI - Accuracy of color duplex ultrasound diagnosis of spontaneous carotid dissection causing ischemia. AB - BACKGROUND AND PURPOSE: Spontaneous dissection of the cervical internal carotid artery (sICAD) is mainly assessed with MRI and magnetic resonance angiography (MRA), which are not always at hand. In contrast, color duplex sonography (CDS) is readily available. We undertook this prospective study to examine the accuracy of CDS to diagnose sICAD in patients with first carotid territory ischemia. METHODS: Consecutive patients with first carotid territory stroke or transient ischemic attack or retinal ischemia underwent clinical and laboratory examinations, ECG, CDS of the cerebral arteries, cranial computed tomography in case of stroke or transient ischemic attack, and echocardiography and 24-hour ECG in selected cases. Patients were included, if they were <65 years of age, CDS showed a probable sICAD (cervical internal carotid artery stenosed or occluded), or had no determined etiology of ischemia. All of the included patients underwent cervical MRI and MRA+/-cerebral catheter angiography. The sonographer was blinded to the results of MRI and angiography studies. RESULTS: We included 177 of 1652 screened patients. Excluded patients (n=1475) were > or =65 years old (n=818), had another determined cause of ischemia (n=1475), and had intracranial hemorrhage (n=58). CDS diagnosed sICAD in 77 of 177 patients, and the etiology of ischemia was undetermined in the remaining 100 patients. Cervical MRI and angiography showed 74 sICAD; there were 6 falsely positive and 3 falsely negative CDS findings. Thus, sensitivity, specificity, and positive and negative predictive values for CDS diagnosis of patients with sICAD causing carotid territory ischemia was 96%, 94%, 92%, and 97%, respectively. CONCLUSIONS: Color duplex ultrasound allows the reliable exclusion of sICAD in patients with carotid territory ischemia, whereas diagnosis of CDS of sICAD must be confirmed with cervical MRI and MRA. PMID- 16373651 TI - Distribution of carotid arterial lesions in Chinese patients with transient monocular blindness. AB - BACKGROUND AND PURPOSE: Asian patients with cerebrovascular diseases have more intracranial atherosclerosis and less extracranial carotid artery stenosis compared with white patients. We systemically evaluated the distribution of carotid arterial lesions in Chinese patients with transient monocular blindness (TMB), which was rarely reported. METHODS: We prospectively evaluated 105 consecutive patients with TMB. All of the patients received ocular and physical examinations, blood tests for coagulation function and autoimmune diseases, and ultrasonography of cervical and intracranial arteries. All of the carotid lesions were confirmed by magnetic resonance angiography or cerebral angiography. RESULTS: Of the 36 (34.3%) patients with significant carotid stenosis (> or =50%), 16 (15.2%) had extracranial carotid stenosis; 17 (16.2%) had carotid siphon stenosis; and 3 (2.9%) had both. The duration, onset, and patterns of visual loss were not different between patients with and without carotid arterial lesion. CONCLUSIONS: This study signified the importance of carotid siphon stenosis as a probable underlying etiology for TMB in Chinese patients. PMID- 16373652 TI - Endovascular recanalization therapy in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: To assess the outcome in acute ischemic stroke patients not eligible for systemic thrombolysis (outside the 3-hour time window, after surgery, or on anticoagulant) undergoing endovascular recanalization therapy (ERT) at the Columbia University Medical Center (CUMC) and to determine US nationwide usage and outcome of ERT in acute ischemic stroke. METHODS: Patients treated at CUMC from 2001 to 2004 and the Nationwide Inpatient Sample (NIS) comprising 20% of all admissions in the United States from 1999 to 2002 were analyzed retrospectively. RESULTS: Thirty-one patients underwent ERT. Mean age was 68+/-14 years, 68% were female, and 45% nonwhite (occlusion sites: internal carotid artery 29%; middle cerebral artery 39%; posterior circulation 32%). Pharmacological or mechanical ERT was initiated beyond 3 hours after symptom onset (median time 4.4 hours) in 61%, 29% had surgery, and 39% were on anticoagulant medication. At discharge, 32% had modified Rankin Scale scores of 0 to 2 (52% discharged home or to rehabilitation facilities); overall mortality was 29%, of which 19% were fatal intracerebral hemorrhages. From the NIS cohort, 477 patients (0.17% of all strokes and 14% of all thrombolysis cases) underwent ERT. Fifteen percent died, and approximately 50% were discharged home or to rehabilitation facilities. Intracerebral hemorrhage occurred in 6%. Fewer good outcomes of the CUMC cohort may be explained by more unfavorable premorbid patient characteristics compared with the NIS cohort. CONCLUSIONS: Despite significant variability in patient characteristics and treatment methods among 2 sources of data analyzed, ERT in stroke patients not eligible for intravenous thrombolysis appears to be a relatively safe and effective treatment alternative that is being used increasingly nationwide. PMID- 16373653 TI - Altered hemodynamics and regional cerebral blood flow in patients with hemodynamically significant stenoses. AB - BACKGROUND AND PURPOSE: Blood oxygen level-dependent (BOLD) contrast largely depends on changes in cerebral blood flow (CBF). Because cerebrovascular disease may result in altered CBF, we assessed the temporal dynamics and magnitude of the BOLD response in patients with major arterial stenoses. METHODS: Seven patients with hemodynamically significant stenoses affecting the anterior circulation (primarily left internal carotid and middle cerebral arteries) were compared with 7 neurologically healthy subjects. Continuous arterial spin-labeled perfusion MRI was used to measure resting CBF globally and within various vascular distributions. The BOLD response was acquired during a visually guided bilateral handball squeeze task while motor performance was recorded by a pressure transducer. RESULTS: Baseline CBF was reduced in bilateral middle cerebral artery and left anterior cerebral artery territories in patients. A prolonged BOLD hemodynamic response was observed in patients in bilateral primary motor cortices but not visual cortex. Patients also exhibited a larger early negative BOLD response, or "initial dip," in left primary motor cortex. There were no differences in motor performance between groups, suggesting behavioral differences were not primarily responsible for the characteristics of the BOLD response. CONCLUSIONS: An initial deoxygenation followed by a delayed hyperemic BOLD response was observed in patients, although resting flow values were not within an ischemic range. A simple visuomotor BOLD activation paradigm can reflect alterations in the hemodynamic response in patients with hemodynamically significant stenoses. PMID- 16373654 TI - Comparison of ABC/2 estimation technique to computer-assisted planimetric analysis in warfarin-related intracerebral parenchymal hemorrhage. AB - BACKGROUND AND PURPOSE: The ABC/2 formula is a reliable estimation technique of intracerebral hematoma volume. However, oral anticoagulant therapy (OAT)-related intracerebral hemorrhage (ICH) compared with primary ICH is based on a different pathophysiological mechanism, and various shapes of hematomas are more likely to occur. Our objective was to validate the ABC/2 technique based on analyses of the hematoma shapes in OAT-related ICH. METHODS: We reviewed the computed tomography scans of 83 patients with OAT-associated intraparenchymal ICH. Location was divided into deep, lobar, cerebellar, and brain stem hemorrhage. Shape of the ICH was divided into (A) round-to-ellipsoid, (B) irregular with frayed margins, and (C) multinodular to separated. The ABC/2 technique was compared with computer assisted planimetric analyses with regard to hematoma site and shape. RESULTS: The mean hematoma volume was 40.83+/-3.9 cm3 (ABC/2) versus 36.6+/-3.5 cm3 (planimetric analysis). Bland-Altman plots suggested equivalence of both estimation techniques, especially for smaller ICH volumes. The most frequent location was a deep hemorrhage (54%), followed by lobar (21%), cerebellar (14%) and brain stem hemorrhage (11%). The most common shape was round-to-ellipsoid (44%), followed by irregular ICH (31%) and separated and multinodular shapes (25%). In the latter, ABC/2 formula significantly overestimated volume by +32.1% (round shapes by +6.7%; irregular shapes by +14.9%; P ANOVA <0.01). Variation of the denominator toward ABC/3 in cases of irregularly and separately shaped hematomas revealed more a precise volume estimation with a deviation of -10.3% in irregular and +5.6% in separately shaped hematomas. CONCLUSIONS: In patients with OAT-related ICH, >50% of bleedings are irregularly shaped. In these cases, hematoma volume is significantly overestimated by the ABC/2 formula. Modification of the denominator to 3 (ie, ABC/3) measured ICH volume more accurately in these patients potentially facilitating treatment decisions. PMID- 16373655 TI - Lessons from nosocomial viral haemorrhagic fever outbreaks. AB - The outbreak of Marburg haemorrhagic fever in Angola in 2004-2005 shows once again the devastating and rapid spread of viral haemorrhagic fevers in medical settings where hygiene practices are poorly applied or ignored. The legacy of years of war and poverty in Angola has resulted in very poor medical education and services. The initial high rate of infection among infants in Angola may have been related to poor hospital practices, possibly administration of vaccines. Though the outbreak in Angola was in a part of Africa not previously known to have filovirus infection, prior ecological modelling had predicted this location and many others. Prevention of future outbreaks will not be easy. The urgent need is dissemination of knowledge and the training, discipline and resources for good clinical practice. Educating the public to demand higher standards could be a powerful tool. Good practices are difficult to establish and maintain on the scale needed. PMID- 16373656 TI - ADAM-9 (MDC-9/meltrin-gamma), a member of the a disintegrin and metalloproteinase family, regulates myeloma-cell-induced interleukin-6 production in osteoblasts by direct interaction with the alpha(v)beta5 integrin. AB - ADAM-9, a member of the a disintegrin and metalloproteinase family, contains both metalloproteinase and disintegrin domains. Myeloma cell lines express ADAM-9; however, its function and role in the pathophysiology of multiple myeloma is unknown. The aim of this study was to establish whether primary myeloma cells express ADAM-9, whether ADAM-9 regulates IL-6 production in human osteoblasts (hOBs), whether ADAM-9 interacts with specific integrin heterodimers, and the identity of downstream signaling pathways. Primary myeloma cells demonstrated increased expression of ADAM-9 (P < .01). ADAM-9 promoted a 5-fold increase in IL 6, but not IL-1beta mRNA, and a dose- and time-dependent increase in IL-6 production by hOBs (P < .01). IL-6 induction was inhibited by an antibody to the alpha(v)beta5 integrin (P < .01) but not by antibodies to other integrin heterodimers. ADAM-9 was shown to bind directly to the alpha(v)beta5 integrin on hOBs. Antibodies to ADAM-9 and alpha(v)beta5 integrin inhibited myeloma cell induced IL-6 production by hOBs (P < .01). Furthermore, inhibitors of p38 MAPK and cPLA2, but not NF-kappaB and JAK2, signaling pathways inhibited ADAM-9 induced IL-6 production by hOBs (P < .01). These data demonstrate that ADAM-9, expressed by myeloma cells, stimulates IL-6 production in hOBs by binding the alpha(v)beta5 integrin. This may have important consequences for the growth and survival of myeloma cells in bone. PMID- 16373657 TI - Relation between JAK2 (V617F) mutation status, granulocyte activation, and constitutive mobilization of CD34+ cells into peripheral blood in myeloproliferative disorders. AB - We studied the relationship between granulocyte JAK2 (V617F) mutation status, circulating CD34(+) cells, and granulocyte activation in myeloproliferative disorders. Quantitative allele-specific polymerase chain reaction (PCR) showed significant differences between various disorders with respect to either the proportion of positive patients (53%-100%) or that of mutant alleles, which overall ranged from 1% to 100%. In polycythemia vera, JAK2 (V617F) was detected in 23 of 25 subjects at diagnosis and in 16 of 16 patients whose disease had evolved into myelofibrosis; median percentages of mutant alleles in these subgroups were significantly different (32% versus 95%, P < .001). Circulating CD34(+) cell counts were variably elevated and associated with disease category and JAK2 (V617F) mutation status. Most patients had granulocyte activation patterns similar to those induced by administration of granulocyte colony stimulating factor. A JAK2 (V617F) gene dosage effect on both CD34(+) cell counts and granulocyte activation was clearly demonstrated in polycythemia vera, where abnormal patterns were mainly found in patients carrying more than 50% mutant alleles. These observations suggest that JAK2 (V617F) may constitutively activate granulocytes and by this means mobilize CD34(+) cells. This exemplifies a novel paradigm in which a somatic gain-of-function mutation is initially responsible for clonal expansion of hematopoietic cells and later for their abnormal trafficking via an activated cell progeny. PMID- 16373658 TI - Clinical implication of centrosome amplification in plasma cell neoplasm. AB - The mechanisms underlying aneuploidy in multiple myeloma (MM) are unclear. Centrosome amplification has been implicated as the cause of chromosomal instability in a variety of tumors and is a potential mechanism causing aneuploidy in MM. Using immunofluorescent (IF) staining, centrosome amplification was detected in 67% of monoclonal gammopathies, including monoclonal gammopathy of undetermined significance (MGUS). We also investigated the gene expression of centrosome proteins. Overall, gene expression data correlated well with IF detected centrosome amplification, allowing us to derive a gene expression-based centrosome index (CI) as a surrogate for centrosome amplification. Clinically, MM patients with high CI (> 4) are associated with poor prognostic genetic and clinical subtypes (chromosome 13 deletion, t(4; 14), t(14;16), and PCLI > 1%, P < .05) and are shown here to have short survival (11.1 months versus 39.1 months, P < .001). On multivariate regression, a high CI is an independent prognostic factor. Given that centrosome amplification is already observed in MGUS and probably integral to early chromosomal instability and myeloma genesis, and patients with more extensive centrosome amplification have shorter survival, the mechanisms leading to centrosome amplification should be investigated because these may offer new avenues for therapeutic intervention. PMID- 16373659 TI - Impact of vaccine-induced mucosal high-avidity CD8+ CTLs in delay of AIDS viral dissemination from mucosa. AB - Natural HIV transmission occurs through mucosa, but it is debated whether mucosal cytotoxic T lymphocytes (CTLs) can prevent or reduce dissemination from the initial mucosal site to the systemic circulation. Also, the role of CTL avidity in mucosal AIDS viral transmission is unknown. To address these questions, we used delay in acute-phase peak viremia after intrarectal challenge as an indicator of systemic dissemination. We found that a peptide-prime/poxviral boost vaccine inducing high levels of high-avidity mucosal CTLs can have an impact on dissemination of intrarectally administered pathogenic SHIV-ku2 in macaques and that such protection correlates better with mucosal than with systemic CTLs and particularly with levels of high-avidity mucosal CTLs. PMID- 16373660 TI - Tumor vascular targeting therapy with viral vectors. AB - Tumor angiogenesis is crucial for the progression and metastasis of cancer. The vasculature of tumor tissue is different from normal vasculature. Therefore, tumor vascular targeting therapy could represent an effective therapeutic strategy with which to suppress both primary tumor growth and tumor metastasis. The use of viral vectors for tumor vascular targeting therapy is a promising strategy based on the unique properties of viral vectors. In order to circumvent the potential problems of antiviral neutralizing antibodies, poor access to extravascular tumor tissue, and toxicities to normal tissue, viral vectors need to be modified to target the tumor endothelial cells. Viral vectors that could be used for tumor vascular targeting therapy include adenoviral vectors, adeno associated viral vectors, retroviral vectors, lentiviral vectors, measles virus, and herpes simplex viral vectors. In this review, we will summarize the strategies available for targeting viral vectors for tumor vascular targeting therapy. PMID- 16373661 TI - Use of all-trans retinoic acid plus arsenic trioxide as an alternative to chemotherapy in untreated acute promyelocytic leukemia. AB - We examined whether combining all-trans retinoic acid (ATRA) and arsenic trioxide (ATO) might be an alternative to ATRA plus chemotherapy in untreated acute promyelocytic leukemia (APL). Twenty-five low-risk patients (white blood cell [WBC] count less than 10 x 10(9)/L [10,000/microL]) received ATRA (45 mg/m(2) daily) and ATO (0.15 mg/kg daily, beginning day 10 of ATRA), and in complete remission (CR) received ATO plus ATRA, without chemotherapy, unless they were reverse transcriptase-polymerase chain reaction (RT-PCR)-positive 3 months from CR date or had molecular relapse. Nineteen high-risk patients were treated identically, but received chemotherapy, generally 9 mg/m(2) gemtuzumab ozogamycin (GO) on day 1 of induction. The CR rate was 39 of 44 (24 of 25 in low-risk, 15 of 19 in high-risk). Disease recurred at 9, 9, and 15 months, respectively, in 3 high-risk patients. The median follow-up time from CR date in the 36 patients alive in first CR is 16 months (15 months in low-risk, 20 months in high-risk), with 9 patients followed for at least 24 months. Each of the 36 patients was PCR negative at last follow-up. Thus, none of the low-risk patients has received chemotherapy, and only 3 high-risk patients (the 3 with relapsed disease) have received chemotherapy past induction. ATRA plus ATO may serve as an alternative to chemotherapy in low-risk untreated APL (eg, in older patients) and, when combined with GO, may improve outcome in high-risk patients. PMID- 16373662 TI - Thalidomide derivative CC-4047 inhibits osteoclast formation by down-regulation of PU.1. AB - CC-4047, an immunomodulatory analog of thalidomide, inhibits multiple myeloma with unknown effects on the human osteoclast lineage. Early osteoclast progenitors are of hematopoietic origin and differentiate into mature bone resorbing multinucleated osteoclasts. We investigated the effects of CC-4047 and thalidomide on human osteoclastogenesis, using in vitro receptor activator of NFkappa-B ligand/macrophage colony-stimulating factor-stimulated bone marrow cell cultures. Treating bone marrow cultures with CC-4047 for 3 weeks decreased osteoclast formation accompanied by complete inhibition of bone resorption. The inhibitory effect was similar when cultures were treated for 3 weeks or for only the first week (90% inhibition), indicating that CC-4047 inhibits early stages of osteoclast formation. Inhibition of osteoclastogenesis by CC-4047 was mediated by a shift of lineage commitment to granulocyte colony-forming units at the expense of granulocyte-macrophage colony-forming units. Further studies revealed that this shift in lineage commitment was mediated through down-regulation of PU.1. Treatment with thalidomide resulted in significantly less potent inhibition of osteoclast formation and bone resorption. These results provide evidence that CC 4047 blocks osteoclast differentiation during early phases of osteoclastogenesis. Therefore, CC-4047 might be a valuable drug for targeting both tumors and osteoclastic activity in patients with multiple myeloma and other diseases associated with osteolytic lesions. PMID- 16373663 TI - Cardiac morbidity and mortality in deferoxamine- or deferiprone-treated patients with thalassemia major. AB - Deferoxamine (DFO) therapy has been associated with improved survival of thalassemia patients. However, cardiac disease remains the main cause of death in those patients. In 1995, the oral chelator deferiprone became available for clinical use. We compared the occurrence of cardiac disease in patients treated only with DFO and in those whose therapy was switched to deferiprone during the period of observation, from January 31, 1995, to December 31, 2003. All patients with thalassemia major treated in 7 Italian centers who were born between 1970 and 1993 and who had not experienced a cardiac event prior to January 1995 were included. DFO only was given to 359 patients, and 157 patients received deferiprone for part of the time. A total of 3,610 patient-years were observed on DFO and 750 on deferiprone. At baseline, the 2 groups were comparable for age and sex, while ferritin levels were significantly higher in patients switched to deferiprone. Fifty-two cardiac events, including 10 cardiac deaths, occurred during therapy with DFO. No cardiac events occurred during deferiprone therapy or within at least 18 months after the end of it. In the setting of a natural history study, deferiprone therapy was associated with significantly greater cardiac protection than deferoxamine in patients with thalassemia major. PMID- 16373664 TI - Lymphomas are sensitive to perforin-dependent cytotoxic pathways despite expression of PI-9 and overexpression of bcl-2. AB - There is considerable interest in immunotherapeutic approaches for lymphoma. The expression of proteinase inhibitor 9 (PI-9), a molecule that inactivates granzyme B, is considered an immune escape mechanism in lymphoma. Further, lymphomas frequently overexpress the antiapoptotic molecule bcl-2, which is able to inhibit perforin-dependent cytotoxic pathways. In this study, the impact of PI-9 and bcl 2 expression on the sensitivity of lymphomas to T- and natural killer (NK) cell mediated cytotoxicity was analyzed. We found PI-9 expression in 10 of 18 lymphoma cell lines and in 9 of 14 primary lymphomas. Overexpression of bcl-2 was found in 8 of 18 cell lines and in 12 of 14 primary lymphomas. All lymphoma cells were sensitive to cytolysis by specific T cells and cytokine-activated NK cells, and no difference in sensitivity was observed with respect to PI-9 or bcl-2 expression. Cytolysis was mediated predominantly through perforin-dependent pathways despite expression of PI-9 and bcl-2. Interestingly, the majority of lymphoma cells were resistant to cytolysis by resting allogeneic NK cells. This was due to the failure of lymphomas to induce degranulation of resting NK cells. These results show that resistance to perforin-dependent pathways is not a relevant immune escape mechanism in lymphoma and therefore is unlikely to impair clinical outcome of immunotherapeutic approaches. PMID- 16373665 TI - Identification of the annexin A2 heterotetramer as a receptor for the plasmin induced signaling in human peripheral monocytes. AB - We have previously demonstrated that plasmin acts as a potent proinflammatory activator of human peripheral monocytes. Here we identify the annexin A2 heterotetramer, composed of annexin A2 and S100A10, as a receptor for the plasmin induced signaling in human monocytes. Monocytes express the annexin A2 heterotetramer on the cell surface as shown by flow cytometry, fluorescence microscopy, and coimmunoprecipitation of biotinylated cell surface proteins. Binding of plasmin to annexin A2 and S100A10 on monocytes was verified by biotin transfer from plasmin labeled with a trifunctional cross-linker. Antibodies directed against annexin A2 or S100A10 inhibited the chemotaxis elicited by plasmin, but not that induced by fMLP. Further, down-regulation of annexin A2 or S100A10 in monocytes by antisense oligodeoxynucleotides impaired the chemotactic response to plasmin, but not that to fMLP. Antisense oligodeoxynucleotides similarly decreased the TNF-alpha release by plasmin-stimulated, but not by LPS stimulated, monocytes. At the molecular level, stimulation with plasmin, but not with catalytically inactivated plasmin, induced cleavage of annexin A2 and dissociation of the heterotetramer complex. Substitution of lysine to alanine in position 27 abolished the cleavage of recombinant annexin A2 in vitro. Together, these data identify the annexin A2 heterotetramer as a signaling receptor activated by plasmin via proteolysis. PMID- 16373666 TI - Regulation of hematopoiesis in vitro and in vivo by invariant NKT cells. AB - Invariant natural killer T cells (iNKT cells) are a small subset of immunoregulatory T cells highly conserved in humans and mice. On activation by glycolipids presented by the MHC-like molecule CD1d, iNKT cells promptly secrete T helper 1 and 2 (Th1/2) cytokines but also cytokines with hematopoietic potential such as GM-CSF. Here, we show that the myeloid clonogenic potential of human hematopoietic progenitors is increased in the presence of glycolipid activated, GM-CSF-secreting NKT cells; conversely, short- and long-term progenitor activity is decreased in the absence of NKT cells, implying regulation of hematopoiesis in both the presence and the absence of immune activation. In accordance with these findings, iNKT-cell-deficient mice display impaired hematopoiesis characterized by peripheral-blood cytopenias, reduced marrow cellularity, lower frequency of hematopoietic stem cells (HSCs), and reduced early and late hematopoietic progenitors. We also show that CD1d is expressed on human HSCs. CD1d-expressing HSCs display short- and long-term clonogenic potential and can present the glycolipid alpha-galactosylceramide to iNKT cells. Thus, iNKT cells emerge as the first subset of regulatory T cells that are required for effective hematopoiesis in both steady-state conditions and under conditions of immune activation. PMID- 16373667 TI - Early breastfeeding cessation in rural Senegal: causes, modes, and consequences. AB - OBJECTIVES: We studied reasons for cessation of breastfeeding before the age of 15 months, replacement feeding modes, and child mortality in West Africa. METHODS: Data were gathered for 12208 children born between 1987 and 1997 in a rural area of Senegal. Interviews were conducted with caregivers of early-weaned children, and child mortality risks were assessed. RESULTS: Fewer than 1% of children had been weaned early. The main reasons for early weaning were maternal death and new pregnancy (in 41% and 27% of cases, respectively). Twenty percent of children had been relactated by a wet nurse, and 16% had received formula. Many early-weaned children died before the age of 2 years (26%), particularly those weaned early as a result of the mother's death (hazard ratio = 5.1; 95% confidence interval [CI] = 1.74, 15.0). Girls had a lower hazard ratio than boys (0.16; 95% CI=0.05, 0.41). CONCLUSIONS: Our results showed that early cessation of breastfeeding was rare but that associated mortality was high, especially when the mother had died. PMID- 16373668 TI - Socioeconomic status and cardiovascular disease among men: the Korean national health service prospective cohort study. AB - OBJECTIVES: We examined the association between socioeconomic status (SES) and myocardial infarction and stroke subtypes, including the possible mediating influence of cardiovascular risk factors. METHODS: We evaluated data on 578756 Korean male public servants aged 30 to 58 years from August 1, 1990, to July 31, 2001. RESULTS: SES had inverse associations with mortality because of myocardial infarction and stroke subtypes, which were not changed by an adjustment for, or stratification by, cardiovascular risk factors. For nonfatal events, SES had positive, null, and inverse associations with myocardial infarction, ischemic stroke, and hemorrhagic stroke, respectively. The association between SES and nonfatal myocardial infarction depended on the presence of risk factors and was positive only among men who had cardiovascular risk factors. Case-fatality after hospital admission for cardiovascular diagnoses was significantly lower among higher SES groups, even after risk factor adjustment. CONCLUSIONS: Inverse SES associations with cardiovascular diseases were not mediated by cardiovascular risk factors among men who were undergoing economic transition. Socioeconomically patterned access to medical care may partly explain these socioeconomic gradients. PMID- 16373669 TI - Zn2+-induced IL-8 expression involves AP-1, JNK, and ERK activities in human airway epithelial cells. AB - Exposure to zinc-laden particulate matter in ambient and occupational settings has been associated with proinflammatory responses in the lung. IL-8 is an important proinflammatory cytokine in the human lung and is induced in human airway epithelial cells exposed to zinc. In this study, we examined the cellular mechanisms responsible for Zn(2+)-induced IL-8 expression. Zn(2+) stimulation resulted in pronounced increases in both IL-8 mRNA and protein expression in the human airway epithelial cell line (BEAS-2B). IL-8 promoter activity was significantly increased by Zn(2+) exposure in BEAS-2B cells, indicating that Zn(2+)-induced IL-8 expression is transcriptionally mediated. Mutation of the activating protein (AP)-1 response element in an IL-8 promoter-enhanced green fluorescent protein construct reduced Zn(2+)-induced IL-8 promoter activity. Moreover, Zn(2+) exposure of BEAS-2B cells induced the phosphorylation of the AP 1 proteins c-Fos and c-Jun. We observed that Zn(2+) exposure induced the phosphorylation of ERK, JNK, and p38 MAPKs, whereas inhibition of ERK or JNK activity blocked IL-8 mRNA and protein expression in BEAS-2B cells treated with Zn(2+). In addition, we investigated the role of protein tyrosine phosphatases in the activation of signaling by Zn(2+). Zn(2+) treatment inhibited ERK- and JNK directed phosphatase activities in BEAS-2B cells. These results suggested that Zn(2+)-induced inhibition of phosphatase activity is an initiating event in MAPK and AP-1 activation that leads to enhanced IL-8 expression by human airway epithelial cells. PMID- 16373670 TI - Increased pulmonary responses to acute ozone exposure in obese db/db mice. AB - Epidemiological studies indicate the incidence of asthma is increased in obese and overweight humans. Responses to ozone (O(3)), an asthma trigger, are increased in obese (ob/ob) mice lacking the satiety hormone leptin. The long form of leptin receptor (Ob-R(b)) is required for satiety; mice lacking this receptor (db/db mice) are also substantially obese. Here, wild-type (WT) and db/db mice were exposed to air or O(3) (2 ppm) for 3 h. Airway responsiveness, measured by the forced oscillation technique, was greater in db/db than WT mice after air exposure. O(3)-induced increases in pulmonary resistance and airway responsiveness were also greater in db/db mice. BALF eotaxin, IL-6, KC, and MIP-2 increased 4 h after O(3) exposure and subsided by 24 h, whereas protein and neutrophils continued to increase through 24 h. For each outcome, the effect of O(3) was significantly greater in db/db than WT mice. Previously published results obtained in ob/ob mice were similar except for O(3)-induced neutrophils and MIP-2, which were not different from WT mice. O(3) also induced pulmonary IL 1beta and TNF-alpha mRNA expression in db/db but not ob/ob mice. Leptin was increased in serum of db/db mice, and pulmonary mRNA expression of short form of leptin receptor (Ob-R(a)) was similar in db/db and WT mice. These data confirm obese mice have innate airway hyperresponsiveness and increased pulmonary responses to O(3). Differences between ob/ob mice, which lack leptin, and db/db mice, which lack Ob-R(b) but not Ob-R(a), suggest leptin, acting through Ob-R(a), can modify some pulmonary responses to O(3). PMID- 16373671 TI - Shedding of soluble ICAM-1 into the alveolar space in murine models of acute lung injury. AB - Intercellular adhesion molecule-1 (ICAM-1; CD54) is an adhesion molecule constitutively expressed in abundance on the cell surface of type I alveolar epithelial cells (AEC) in the normal lung and is a critical participant in pulmonary innate immunity. At many sites, ICAM-1 is shed from the cell surface as a soluble molecule (sICAM-1). Limited information is available regarding the presence, source, or significance of sICAM-1 in the alveolar lining fluid of normal or injured lungs. We found sICAM-1 in the bronchoalveolar lavage (BAL) fluid of normal mice (386 +/- 50 ng/ml). Additionally, sICAM-1 was spontaneously released by murine AEC in primary culture as type II cells spread and assumed characteristics of type I cells. Shedding of sICAM-1 increased significantly at later points in culture (5-7 days) compared with earlier time points (3-5 days). In contrast, treatment of AEC with inflammatory cytokines had limited effect on sICAM-1 shedding. BAL sICAM-1 was evaluated in in vivo models of acute lung injury. In hyperoxic lung injury, a reversible process with a major component of leak across the alveolar wall, BAL fluid sICAM-1 only increased in parallel with increased alveolar protein. However, in lung injury due to FITC, there were increased levels of sICAM-1 in BAL that were independent of changes in BAL total protein concentration. We speculate that after lung injury, changes in sICAM-1 in BAL fluid are associated with progressive injury and may be a reflection of type I cell differentiation during reepithelialization of the injured lung. PMID- 16373672 TI - The neuropeptide neuromedin U activates eosinophils and is involved in allergen induced eosinophilia. AB - Neuromedin U (NMU) is a neuropeptide expressed not only in the central nervous system but also in various organs, including the gastrointestinal tract and lungs. NMU interacts with two G protein-coupled receptors, NMU-R1 and NMU-R2. Although NMU-R2 is expressed in a specific region of the brain, NMU-R1 is expressed in various peripheral tissues, including immune and hematopoietic cells. Our recent study demonstrated an important role of NMU in mast cell mediated inflammation. In this study, we showed that airway eosinophilia was reduced in NMU-deficient mice in an allergen-induced asthma model. There were no differences in the antigen-induced Th2 responses between wild-type and NMU knockout mice. NMU-R1 was highly expressed in the eosinophil cell line, and NMU directly induced Ca(2+) mobilization and extracellular/signal-regulated kinase phosphorylation. NMU also induced cell adhesion to components of the extracellular matrix (fibronectin and collagen type I), and chemotaxis in vitro. Furthermore, NMU-R1 was also expressed in human peripheral blood eosinophils, and NMU induced cell adhesion in a dose-dependent manner. These data indicate that NMU promotes eosinophil infiltration into inflammatory sites by directly activating eosinophils. Our study suggests that NMU receptor antagonists could be novel targets for pharmacological inhibition of allergic inflammatory diseases, including asthma. PMID- 16373673 TI - Regulated expression of hypoxia-inducible factors during postnatal and postpneumonectomy lung growth. AB - We previously found increased expression of erythropoietin receptor (EPO-R) in peripheral dog lung during postnatal and postpneumonectomy (PNX) lung growth. To study the upstream regulation of EPO-R, we analyzed the expression of hypoxia inducible factors (HIF)-1alpha, -2alpha, and -3alpha during postnatal lung growth in immature and mature (2.5 and 12 mo old, respectively) dogs and during compensatory lung growth 3 wk and 10 mo after right PNX. Relative to their respective controls, HIF-1alpha transcript was 52-95% higher in immature lungs and 284% higher in the remaining lung 3 wk post-PNX. HIF-2alpha transcript did not change during maturation but was 42% lower 3 wk post-PNX. HIF-3alpha transcript was 53-65% lower in both the immature lung and 3 wk post-PNX. Changes were no longer detectable 10 mo post-PNX. No change in HIF transcripts was observed in kidney and liver post-PNX. Consistent with the mRNA changes, HIF 1alpha protein was 120 and 196% higher in growing lungs and 3 wk post-PNX relative to their respective controls. Overexpression of HIF-1alpha in cultured HEK-293 cells increased endogenous expression of EPO-R protein. These results demonstrate regulated expression of the HIF system and parallel changes in HIF 1alpha and EPO-R expression during two types of lung growth. Because the normal growing lung is not hypoxic, the HIF system likely responds to other signals encountered during sustained lung strain. PMID- 16373674 TI - Microtensile testing, nanoleakage, and biodegradation of resin-dentin bonds. PMID- 16373675 TI - Diabetes-enhanced inflammation and apoptosis--impact on periodontal pathology. AB - Diabetes, particularly type 2 diabetes, is a looming health issue with many ramifications. Because diabetes alters the cellular microenvironment in many different types of tissues, it causes myriad untoward effects, collectively referred to as 'diabetic complications'. Two cellular processes affected by diabetes are inflammation and apoptosis. This review discusses how diabetes enhanced inflammation and apoptosis may affect the oral environment. In particular, dysregulation of tumor necrosis factor and the formation of advanced glycation products, both of which occur at higher levels in diabetic humans and animal models, potentiate inflammatory responses and induce apoptosis of matrix producing cells. The enhanced loss of fibroblasts and osteoblasts through apoptosis in diabetics could contribute to limited repair of injured tissue, particularly when combined with other known deficits in diabetic wound-healing. These findings may shed light on diabetes-enhanced risk of periodontal diseases. PMID- 16373676 TI - The role of matrix metalloproteinases (MMPs) in human caries. AB - The objective of this review is to summarize our understanding of the role of host matrix metalloproteinases (MMPs) in the caries process and to discuss new therapeutic avenues. MMPs hydrolyze components of the extracellular matrix and play a central role in many biological and pathological processes. MMPs have been suggested to play an important role in the destruction of dentin organic matrix following demineralization by bacterial acids and, therefore, in the control or progression of carious decay. Host-derived MMPs can originate both from saliva and from dentin. They may be activated by an acidic pH brought about by lactate release from cariogenic bacteria. Once activated, they are able to digest demineralized dentin matrix after pH neutralization by salivary buffers. Furthermore, the degradation of SIBLINGs (Small Integrin-binding Ligand N-linked Glycoproteins) by the caries process may potentially enhance the release of MMPs and their activation. This review also explores the different available MMP inhibitors, natural or synthetic, and suggests that MMP inhibition by several inhibitors, particularly by natural substances, could provide a potential therapeutic pathway to limit caries progression in dentin. PMID- 16373677 TI - Structural and mechanical properties of mandibular condylar bone. AB - The trabecular bone of the mandibular condyle is structurally anisotropic and heterogeneous. We hypothesized that its apparent elastic moduli are also anisotropic and heterogeneous, and depend on trabecular density and orientation. Eleven condyles were scanned with a micro-CT system. Volumes of interest were selected for the construction of finite element models. We simulated compressive and shear tests to determine the principal mechanical directions and the apparent elastic moduli. Compressive moduli were relatively large in directions acting in the sagittal plane, and small in the mediolateral direction. The degree of mechanical anisotropy ranged from 4.7 to 10.8. Shear moduli were largest in the sagittal plane and smallest in the transverse plane. The magnitudes of the moduli varied with the condylar region and were proportional to the bone volume fraction. Furthermore, principal mechanical direction correlated significantly with principal structural direction. It was concluded that variation in trabecular structure coincides with variation in apparent mechanical properties. PMID- 16373678 TI - Polymerization kinetics of pre-heated composite. AB - Temperature affects the polymerization behavior of dimethacrylate-based materials. This study describes the influence of pre-polymerization temperature and exposure duration on polymerization kinetics of a commercial dental photo activated composite at the top and at 2-mm depth. We used the temperature controlled stage of a diamond-attenuated-total-reflectance unit to pre-set composite temperature between 3 degrees and 60 degrees C. Composite was light exposed by a conventional quartz-tungsten-halogen curing unit for 5, 10, 20, or 40 sec. Real-time conversion, maximum conversion rate (R(p)(max)), time to achieve R(p)(max), and conversion at R(p)(max) were calculated from infrared spectra. Composite pre-warming enhanced maximal polymerization rate and overall monomer conversion (top significantly greater than 2 mm). Time when R(p)(max) occurred did not change with temperature, but occurred sooner at the top than at 2-mm depth. Conversion at R(p)(max) increased with temperature, allowing more of the reaction to occur prior to vitrification than at room temperature. PMID- 16373679 TI - Polyelectrolyte multilayer film coating and stability at the surfaces of oral prosthesis base polymers: an in vitro and in vivo study. AB - A new type of coating involving a layer-by-layer technique has been recently reported. This coating is composed of a polyelectrolyte multilayer film that confers specific properties on surfaces to which it is applied. Here, we studied the applicability of such a technique to the coating of oral prostheses, by first testing the construction of polyelectrolyte multilayer films on several polymers used in oral prosthesis bases, and, subsequently, by studying the stability of these coatings in vitro, in human saliva, and in vivo in a rat model. We demonstrated that the multilayered films are able to coat the surfaces of all tested polymers completely, thus increasing their wettability. We also showed that saliva does not degrade the film after 7 days in vitro and after 4 days in vivo. Taken together, our results establish that the layer-by-layer technique is suitable for the coating of oral devices. PMID- 16373680 TI - Trigeminal nociceptors express TLR-4 and CD14: a mechanism for pain due to infection. AB - Although certain bacterial species appear to be risk factors for pain due to odontogenic infections, comparatively little is known about the potential mechanisms mediating this effect. In this study, we tested the hypothesis that trigeminal nociceptive neurons express the TLR4 or CD14 receptors, thus enabling sensory neurons to detect and respond to tissue levels of bacterial substances such as lipopolysaccharide (LPS). Immunohistochemical analyses of human and rat trigeminal neurons demonstrated that a capsaicin-sensitive subclass of nociceptors (defined by expression of TRPV1, a capsaicin receptor) expresses both TLR4 and CD14. Moreover, human dental pulp collected from patients with caries lesions demonstrated co-localization of TLR4 and CD14, with markers of peripheral sensory neurons. Collectively, these studies indicate that the capsaicin sensitive subclass of trigeminal nociceptors expresses TLR4 and CD14. These results indicate that pain due to bacterial infections may result, in part, from direct activation of nociceptors by bacterial products such as LPS. PMID- 16373681 TI - Influence of biosurfactant on interactive forces between mutans Streptococci and enamel measured by atomic force microscopy. AB - Although interactive forces, influenced by environmental conditions, between oral bacteria and tooth surfaces are important for the development of plaque, they have never been estimated. It is hypothesized that interactive forces, as measured by atomic force microscopy, between enamel with or without a pellicle and two strains of mutans streptococci become less attractive by the application of a Streptococcus mitis BMS biosurfactant coating. Upon approach of each of the strains toward bare and pellicle-coated enamel, adsorbed biosurfactant increased the range of the repulsive forces. Upon retraction of the enamel surface, small adhesion forces (0.8-0.9 nN) were measured for bare enamel that almost disappeared after biosurfactant coating. The prevalence and magnitude of the adhesion forces also decreased upon pellicle-coating of the enamel, with a minor effect of adsorbed biosurfactant. These findings indicate that adsorbed S. mitis BMS biosurfactant changes the interactive forces between the mutans streptococci studied and enamel, explaining the effects of biosurfactant on adhesion. PMID- 16373682 TI - Gingival tissue and crevicular fluid co-operation in adult periodontitis. AB - Activated matrix metalloproteinase-3 (MMP-3) can contribute to periodontal ligament destruction in adult periodontitis. Since MMP-3 has been reported to activate proMMP-8 and -9, it was speculated that gingival tissue fibroblast derived MMP-3 might, in periodontitis, be responsible for activation of gingival crevicular fluid (GCF) neutrophil-derived proMMP-8 and -9. Immunohistochemistry disclosed MMP-3 in gingival fibroblasts in periodontitis. Cultured gingival fibroblasts released only pro-MMP-3 when stimulated with tumor necrosis factor alpha. However, Western blot revealed partially activated MMP-3, MMP-8, and MMP-9 in periodontitis GCF. Active MMP-8 (p < 0.05) and MMP-9 (p < 0.05) correlated with the presence of active MMP-3. It seems that resident gingival fibroblasts produce pro-MMP-3 in GCF, where it becomes activated, probably by cathepsin G or elastase released by neutrophils. Active MMP-3 then activates neutrophil-derived pro-MMP-8 and -9. Different tissue compartments/cells exert co-operative actions in mutual local MMP activation cascades. PMID- 16373683 TI - Pilocarpine-induced salivation and thirst in conscious rats. AB - The muscarinic receptor agonist pilocarpine is widely used as a sialogogue. It has been well-established that it also induces water intake in animals. However, the mechanisms underlying the relationships between these events are unknown. To address this problem, we examined water intake and parotid salivary secretion in conscious rats. Intraperitoneally injected pilocarpine increased both water intake and salivary secretion. Intracerebroventricularly injected pilocarpine also induced water intake, but not salivary secretion. Intracerebroventricularly applied atropine, a muscarinic receptor antagonist, suppressed the water intake produced by pilocarpine applied intraperitoneally and intracerebroventricularly. However, it did not affect the salivary secretion induced by pilocarpine applied peripherally. We conclude that peripherally applied pilocarpine affects the parotid glands and the thirst center in the central nervous system, while it may induce salivary secretion mainly via peripheral responses, but water intake mainly via the central nervous system. PMID- 16373684 TI - Overall activity of all masticatory muscles during lateral excursion. AB - Quantification of the overall activity of every masticatory muscle is requisite for the analysis of stomatognathic function, which has not been accomplished by conventional electromyography. We used positron emission tomography and 18F fluoro-deoxy-glucose to quantify the overall activity of every masticatory muscle during lateral excursion, and to evaluate the relative contribution of each masticatory muscle to lateral excursion. The present study suggested that lateral and medial pterygoid muscles are more responsible for lateral excursion than are masseter and temporal muscles. In particular, the contralateral lateral pterygoid muscle plays a major role, followed by the contralateral medial pterygoid muscle. PMID- 16373685 TI - Full-mouth tooth extraction lowers systemic inflammatory and thrombotic markers of cardiovascular risk. AB - Prior studies of a link between periodontal and cardiovascular disease have been limited by being predominantly observational. We used a treatment intervention model to study the relationship between periodontitis and systemic inflammatory and thrombotic cardiovascular indicators of risk. We studied 67 adults with advanced periodontitis requiring full-mouth tooth extraction. Blood samples were obtained: (1) at initial presentation, immediately prior to treatment of presenting symptoms; (2) one to two weeks later, before all teeth were removed; and (3) 12 weeks after full-mouth tooth extraction. After full-mouth tooth extraction, there was a significant decrease in C-reactive protein, plasminogen activator inhibitor-1 and fibrinogen, and white cell and platelet counts. This study shows that elimination of advanced periodontitis by full-mouth tooth extraction reduces systemic inflammatory and thrombotic markers of cardiovascular risk. Analysis of the data supports the hypothesis that treatment of periodontal disease may lower cardiovascular risk, and provides a rationale for further randomized studies. PMID- 16373686 TI - Welfare dental intervention improves employment and quality of life. AB - Chronic, untreated oral disease adversely affects one's systemic health, quality of life, and economic productivity. This study evaluated the effect of rehabilitative dental treatment on the oral-health-related quality of life and employment of welfare recipients. Three hundred and seventy-seven participants in a novel welfare dental program received oral examinations, questionnaires, and rehabilitative dental treatment. Seventy-nine percent of participants exhibited improvement in their oral-health-related quality-of-life scores following dental treatment. Improved OHIP-14 change scores were associated with being Caucasian or African-American, initial poor general health, severity of treatment urgency, worse baseline oral-health-related quality-of-life scores, subsequent patient satisfaction with the Dental Program, and resolution of their chief complaint (all p < 0.04). Those who completed their dental treatment were twice as likely to achieve a favorable/neutral employment outcome (OR = 2.01, 95%CI = 1.12, 3.62). Thus, oral health improved the quality of life and employment outcome for this welfare population. PMID- 16373687 TI - Risk factors of early childhood caries in a Southeast Asian population. AB - A retrospective cohort study on ECC and associated factors was conducted among mothers with 25- to 30-month-old infants in a community where prolonged breastfeeding was common practice. All infants who consumed sugary supplementary food or rice that was pre-chewed by the mother, or who fell asleep with the breast nipple in their mouths, had ECC. Infants without those habits, and who were breastfed up to 12 months, had no ECC. Breastfeeding during the day beyond the age of 12 months was not associated with ECC, but infants who were breastfed at night > 2 times had an OR for ECC of 35 (CI 6-186), and those who were exposed to > 15 min per nocturnal feeding had an OR for ECC of 100 (CI 10-995). The present study indicates that, in this population, besides the consumption of sugars and pre-chewed rice, nocturnal breastfeeding after the age of 12 months poses a risk of developing ECC. PMID- 16373688 TI - Radiographic assessment of dental health in middle-aged men following sudden cardiac death. AB - Poor oral health has been suggested to be a risk factor for myocardial infarction. To study if dental pathology might predispose to pre-hospital sudden cardiac death, and using a sum index of panoramic tomography findings, we compared the oral health of middle-aged (33-69 yrs) male victims (Helsinki Sudden Death Study) of sudden cardiac death (n = 117) with that of controls, who died of non-cardiac diseases (n = 63) or suffered unnatural sudden death (n = 120). The mean number of teeth was 15.2, and 17.4% of the men were edentulous. Frequent age associated findings in dentate victims were fillings (79.9%), horizontal bone loss (72.1%), periapical lesions (45.6%), residual roots (38.2%), and vertical pockets (30.9%). In multivariate analysis with coronary heart disease risk factors and number of teeth as covariates, poor oral health was associated (p = 0.053) with the risk of sudden cardiac death along with age, smoking, and body mass index. This association was especially strong (p = 0.009) among victims < 50 yrs. PMID- 16373689 TI - A novel cyclohexene derivative, ethyl (6R)-6-[N-(2-Chloro-4 fluorophenyl)sulfamoyl]cyclohex-1-ene-1-carboxylate (TAK-242), selectively inhibits toll-like receptor 4-mediated cytokine production through suppression of intracellular signaling. AB - Proinflammatory mediators such as cytokines and NO play pivotal roles in various inflammatory diseases. To combat inflammatory diseases successfully, regulation of proinflammatory mediator production would be a critical process. In the present study, we investigated the in vitro effects of ethyl (6R)-6-[N-(2-chloro 4-fluorophenyl)sulfamoyl]cyclohex-1-ene-1-carboxylate (TAK-242), a novel small molecule cytokine production inhibitor, and its mechanism of action. In RAW264.7 cells and mouse peritoneal macrophages, TAK-242 suppressed lipopolysaccharide (LPS)-induced production of NO, tumor necrosis factor-alpha (TNF-alpha), and interleukin (IL)-6, with 50% inhibitory concentration (IC50) of 1.1 to 11 nM. TAK 242 also suppressed the production of these cytokines from LPS-stimulated human peripheral blood mononuclear cells (PBMCs) at IC50 values from 11 to 33 nM. In addition, the inhibitory effects on the LPS-induced IL-6 and IL-12 production were similar in human PBMCs, monocytes, and macrophages. TAK-242 inhibited mRNA expression of IL-6 and TNF-alpha induced by LPS and interferon-gamma in RAW264.7 cells. The phosphorylation of mitogen-activated protein kinases induced by LPS was also inhibited in a concentration-dependent manner. However, TAK-242 did not antagonize the binding of LPS to the cells. It is noteworthy that TAK-242 suppressed the cytokine production induced by Toll-like receptor (TLR) 4 ligands, but not by ligands for TLR2, -3, and -9. In addition, IL-1beta-induced IL-8 production from human PBMCs was not markedly affected by TAK-242. These data suggest that TAK-242 suppresses the production of multiple cytokines by selectively inhibiting TLR4 intracellular signaling. Finally, TAK-242 is a novel small molecule TLR4 signaling inhibitor and could be a promising therapeutic agent for inflammatory diseases, whose pathogenesis involves TLR4. PMID- 16373690 TI - Differential mRNA processing in hematopoietic stem cells. AB - Hematopoietic stem cells (HSCs) maintain tissue homeostasis by rapidly responding to environmental changes. Although this function is well understood, the molecular mechanisms governing this characteristic are largely unknown. We used a sequenced-based strategy to explore the role of both transcriptional and post transcriptional regulation in HSC biology. We characterized the gene expression differences between HSCs, both quiescent and proliferating, and their differentiated progeny. This analysis revealed a large fraction of sequence tags aligned to intronic sequences, which we showed were derived from unspliced transcripts. A comparison of the biological properties of the observed spliced versus unspliced transcripts in HSCs showed that the unspliced transcripts were enriched in genes involved in DNA binding and RNA processing. In addition, levels of unspliced message decreased in a transcript-specific fashion after HSC activation in vivo. This change in unspliced transcript level coordinated with increases in gene expression of splicing machinery components. Combined, these results suggest that post-transcriptional regulation is important in HSC activation in vivo. PMID- 16373691 TI - Anatomical compartments modify the response of human hematopoietic cells to a mitogenic signal. AB - Methods for specifically regulating transplanted cells have many applications in gene and cell therapy. We examined the response of human cord blood CD34+ cells to a specific mitotic signal in vivo. Using a conditional signaling molecule (F36VMpl) that is specifically activated by an artificial ligand called a chemical inducer of dimerization (CID), human hematopoietic cells transplanted into immune deficient mice were induced to proliferate. Only differentiating erythroid precursors and multipotential and erythroid progenitors (colony-forming unit [CFU]-mix and burst forming unitserythroid [BFUe]) responded; however, the nature of the response differed markedly between bone marrow and spleen. In the marrow, F36VMpl induced a 12- to 17-fold expansion of differentiated erythroid precursors and a loss of CFU-mix and BFUe. In the spleen, F36VMpl induced a marked rise in BFUe and CFU-mix and, relative to marrow, a much less prominent rise in more mature red cells. Clonal analysis was most consistent with the interpretation that the spleen and bone marrow differentially regulate the response of human progenitors to a mitotic signal, possibly influencing progenitor expansion versus differentiation. These findings establish CIDs as in vivo growth factors for human hematopoietic cells. PMID- 16373692 TI - Thymidine analogs are transferred from prelabeled donor to host cells in the central nervous system after transplantation: a word of caution. AB - Thymidine analogs, including bromodeoxyuridine, chlorodeoxyuridine, iododeoxyuridine, and tritiated thymidine, label dividing cells by incorporating into DNA during S phase of cell division and are widely employed to identify cells transplanted into the central nervous system. However, the potential for transfer of thymidine analogs from grafted cells to dividing host cells has not been thoroughly tested. We here demonstrate that graft-derived thymidine analogs can become incorporated into host neural precursors and glia. Large numbers of labeled neurons and glia were found 3-12 weeks after transplantation of thymidine analog-labeled live stem cells, suggesting differentiation of grafted cells. Remarkably, however, similar results were obtained after transplantation of dead cells or labeled fibroblasts. Our findings reveal for the first time that thymidine analog labeling may not be a reliable means of identifying transplanted cells, particularly in highly proliferative environments such as the developing, neurogenic, or injured brain. PMID- 16373693 TI - Differential expression of alpha2 integrin separates long-term and short-term reconstituting Lin-/loThy1.1(lo)c-kit+ Sca-1+ hematopoietic stem cells. AB - Self-renewing, multipotent hematopoietic stem cells are highly enriched within the Lin- Thy1.1(lo)c-kit+ Sca-1+ subset of mouse bone marrow. However, heterogeneous expression within this population of certain cell surface markers raises the possibility that it may be further fractionated phenotypically and perhaps functionally. We previously identified alpha2-integrin (CD49b) as a surface marker with heterogeneous expression on Lin(-/lo)Thy1.1(lo)c-kit+ Sca-1+ stem cells. To determine whether differences in alpha2 expression were indicative of differences in stem cell function, we purified alpha2- and alpha2hi stem cells by fluorescence-activated cell sorting and analyzed their function in long- and short-term hematopoietic reconstitution assays. Both alpha2- and alpha2hi cells could give rise to mature lymphoid and myeloid cells after transplantation into lethally irradiated congenic recipients. However, alpha2hi cells supported hematopoiesis for only a short time (<4 weeks), whereas alpha2- cells reproducibly yielded robust, long-term (>20 weeks) reconstitution, suggesting that alpha2- cells represent a more primitive population than do alpha2hi cells. Consistent with this idea, alpha2- Lin(-/lo)Thy1.1(lo)c-kit+ Sca-1+ cells exhibited an approximately sixfold decreased frequency of spleen colony-forming units (day 12) versus alpha2hi cells. Furthermore, bone marrow cells isolated from animals transplanted >20 weeks previously with 20 alpha2- Lin( /lo)Thy1.1(lo)c-kit+ Sca-1+ cells included both alpha2- and alpha2hi stem cells of donor origin, indicating that alpha2hi cells are likely lineal descendents of alpha2- cells. Interestingly, alpha2 integrin expression is significantly reduced on lineage-restricted oligopotent progenitors in the marrow, suggesting that high level expression of alpha2 selectively marks a subset of primitive hematopoietic cells which retains multilineage reconstitution potential but exhibits reduced self-renewal capacity. PMID- 16373694 TI - Characterization and multipotentiality of human fetal femur-derived cells: implications for skeletal tissue regeneration. AB - To date, the plasticity, multipotentiality, and characteristics of progenitor cells from fetal skeletal tissue remain poorly defined. This study has examined cell populations from human fetal femurs in comparison with adult-derived mesenchymal cell populations. Real-time quantitative polymerase chain reaction demonstrated expression of mesenchymal progenitor cell markers by fetal-derived cells in comparison with unselected adult-derived and immunoselected STRO-1 enriched adult populations. Multipotentiality was examined using cells derived from femurs and single-cell clones, culture-expanded from explants, and maintained in basal medium prior to exposure to adipogenic, osteogenic, and chondrogenic conditions. Adipocyte formation was confirmed by Oil Red O lipid staining and aP2 immunocytochemistry, with expression of peroxisome proliferation activated receptor-gamma detected only in adipogenic conditions. In chondrogenic pellets, chondrocytes lodged within lacunae and embedded within dense proteoglycan matrix were observed using Alcian blue/Sirius red staining and type II collagen immunocytochemistry. Osteogenic differentiation was confirmed by alkaline phosphatase staining and type I collagen immunocytochemistry as well as by gene expression of osteopontin and osteocalcin. Single-cell clonal analysis was used to demonstrate multipotentiality of the fetal-derived populations with the formation of adipogenic, chondrogenic, and osteogenic populations. Mineralization and osteoid formation were observed after culture on biomimetic scaffolds with extensive matrix accumulation both in vitro and in vivo after subcutaneous implantation in severely compromised immunodeficient mice. These studies demonstrate the proliferative and multipotential properties of fetal femur-derived cells in comparison with adult-derived cells. Selective differentiation and immunophenotyping will determine the potential of these fetal cells as a unique alternative model and cell source in the restoration of damaged tissue. PMID- 16373695 TI - Multipotent neural stem cells from the adult tegmentum with dopaminergic potential develop essential properties of functional neurons. AB - Neurogenesis in the adult brain occurs within the two principal neurogenic regions: the hippocampus and the subventricular zone of the lateral ventricles. The occurrence of adult neurogenesis in non-neurogenic regions, including the midbrain, remains controversial, but isolation of neural stem cells (NSCs) from several parts of the adult brain, including the substantia nigra, has been reported. Nevertheless, it is unclear whether adult NSCs do have the capacity to produce functional dopaminergic neurons, the cell type lost in Parkinson's disease. Here, we describe the isolation, expansion, and in vitro characterization of adult mouse tegmental NSCs (tNSCs) and their differentiation into functional nerve cells, including dopaminergic neurons. These tNSCs showed neurosphere formation and expressed high levels of early neuroectodermal markers, such as the proneural genes NeuroD1, Neurog2, and Olig2, the NSC markers Nestin and Musashi1, and the proliferation markers Ki67 and BrdU (5-bromo-2 deoxyuridine). The cells showed typical propidium iodide-fluorescence-activated cell sorting analysis of slowly dividing cells. In the presence of selected growth factors, tNSCs differentiated into astroglia, oligodendroglia, and neurons expressing markers for cholinergic, GABAergic, and glutamatergic cells. Electrophysiological analyses revealed functional properties of mature nerve cells, such as tetrodotoxin-sensitive sodium channels, action potentials, as well as currents induced by GABA (gamma-aminobutyric acid), glutamate, and NMDA (N methyl-D-aspartate). Clonal analysis demonstrated that individual NSCs retain the capacity to generate both glia and neurons. After a multistep differentiation protocol using co-culture conditions with PA6 stromal cells, a small number of cells acquired morphological and functional properties of dopaminergic neurons in culture. Here, we demonstrate the existence of adult tNSCs with functional neurogenic and dopaminergic potential, a prerequisite for future endogenous cell replacement strategies in Parkinson's disease. PMID- 16373696 TI - Molecular profile and partial functional analysis of novel endothelial cell derived growth factors that regulate hematopoiesis. AB - Recent progress has been made in the identification of the osteoblastic cellular niche for hematopoietic stem cells (HSCs) within the bone marrow (BM). Attempts to identify the soluble factors that regulate HSC self-renewal have been less successful. We have demonstrated that primary human brain endothelial cells (HUBECs) support the ex vivo amplification of primitive human BM and cord blood cells capable of repopulating non-obese diabetic/severe combined immunodeficient repopulating (SCID) mice (SCID repopulating cells [SRCs]). In this study, we sought to characterize the soluble hematopoietic activity produced by HUBECs and to identify the growth factors secreted by HUBECs that contribute to this HSC supportive effect. Extended noncontact HUBEC cultures supported an eight-fold increase in SRCs when combined with thrombopoietin, stem cell factor, and Flt-3 ligand compared with input CD34(+) cells or cytokines alone. Gene expression analysis of HUBEC biological replicates identified 65 differentially expressed, nonredundant transcripts without annotated hematopoietic activity. Gene ontology studies of the HUBEC transcriptome revealed a high concentration of genes encoding extracellular proteins with cell-cell signaling function. Functional analyses demonstrated that adrenomedullin, a vasodilatory hormone, synergized with stem cell factor and Flt-3 ligand to induce the proliferation of primitive human CD34(+)CD38(-)lin(-) cells and promoted the expansion of CD34(+) progenitors in culture. These data demonstrate the potential of primary HUBECs as a reservoir for the discovery of novel secreted proteins that regulate human hematopoiesis. PMID- 16373697 TI - Targeting stathmin in prostate cancer. AB - Stathmin is the founding member of a family of microtubule-destabilizing proteins that regulate the dynamics of microtubule polymerization and depolymerization. Stathmin is expressed at high levels in a variety of human cancers and provides an attractive molecule to target in cancer therapies that disrupt the mitotic apparatus. We developed replication-deficient bicistronic adenoviral vectors that coexpress green fluorescent protein and ribozymes that target stathmin mRNA. The therapeutic potential of these recombinant adenoviruses was tested in an experimental androgen-independent LNCaP prostate cancer model. Adenovirus mediated transfer of anti-stathmin ribozymes resulted in efficient transduction and marked inhibition of stathmin expression in these cells. Cells that were transduced with the anti-stathmin adenoviruses showed a dramatic dose-dependent growth inhibition. This was associated with accumulation of LNCaP cells in the G2 M phases of the cell cycle. A similar dose-dependent inhibition of clonogenic potential was also observed in cells infected with anti-stathmin adenoviruses. Morphologic and biochemical analysis of infected cells showed a marked increase in apoptosis characterized by detachment of the cells, increased chromatin condensation, activation of caspase-3, and fragmentation of internucleosomal DNA. If these findings are confirmed in vivo, it may provide an effective approach for the treatment of prostate cancer. PMID- 16373698 TI - A dityrosyl-diiron radical cofactor center is essential for human ribonucleotide reductases. AB - Ribonucleotide reductase catalyzes the reduction of ribonucleotides to deoxyribonucleotides for DNA biosynthesis. A tyrosine residue in the small subunit of class I ribonucleotide reductase harbors a stable radical, which plays a central role in the catalysis process. We have discovered that an additional tyrosine residue, conserved in human small subunits hRRM2 and p53R2, is required for the radical formation and enzyme activity. Mutations of this newly identified tyrosine residue obliterated the stable radical and the enzymatic activity of human ribonucleotide reductases shown by electron paramagnetic resonance spectroscopy and enzyme activity assays. Three-dimensional structural analysis reveals for the first time that these two tyrosines are located at opposite sides of the diiron cluster. We conclude that both tyrosines are necessary in maintaining the diiron cluster of the enzymes, suggesting that the assembly of a dityrosyl-diiron radical cofactor center in human ribonucleotide reductases is essential for enzyme catalytic activity. These results should provide insights to design better ribonucleotide reductase inhibitors for cancer therapy. PMID- 16373699 TI - Knockdown of the cytoprotective chaperone, clusterin, chemosensitizes human breast cancer cells both in vitro and in vivo. AB - Clusterin is a stress-associated cytoprotective chaperone up-regulated by various apoptotic triggers in many cancers and confers treatment resistance when overexpressed. The objectives of this study were to evaluate clusterin expression levels in human breast cancer and to determine whether antisense oligonucleotides or double-stranded small interfering RNAs (siRNA) targeting the clusterin gene enhance apoptosis induced by paclitaxel. Clusterin immunostaining was evaluated in a tissue microarray of 379 spotted breast cancers. The effect of hormone withdrawal, paclitaxel treatment, clusterin antisense oligonucleotide (OGX-011), and siRNA treatments on clusterin expression was examined in MCF-7 and MDA-MB-231 cells. Northern, quantitative real-time PCR, and Western analyses were used to measure change in clusterin mRNA and protein levels. The effect of OGX-011 or siRNA clusterin treatment on chemosensitivity to paclitaxel was done in both cell lines in vitro, whereas the ability of OGX-011 to chemosensitize in vivo was evaluated in athymic mice bearing MCF-7 tumors. Clusterin was expressed in 62.5% of tumors within the tissue microarray. Clusterin expression increased after estrogen withdrawal and paclitaxel treatment in vitro in MCF-7 cells. OGX-011 or siRNA clusterin decreased clusterin levels by >90% in a dose-dependent, sequence specific manner and significantly enhanced chemosensitivity to paclitaxel in vitro. When combined, OGX-011 or siRNA clusterin reduced the IC50 by 2-log compared with paclitaxel alone. In vivo administration of OGX-011 enhanced the effects of paclitaxel to significantly delay MCF-7 tumor growth. These data identify clusterin as a valid therapeutic target and provides preclinical proof of-principle to test OGX-011 in multimodality therapies for breast cancer. PMID- 16373700 TI - Transcriptional targeting modalities in breast cancer gene therapy using adenovirus vectors controlled by alpha-lactalbumin promoter. AB - The breast-specific antigen alpha-lactalbumin is expressed in >60% of breast cancer tissues. To evaluate the effect of gene therapy for breast cancer by controlling adenovirus replication with human alpha-lactalbumin promoter, we investigated the activity of a 762-bp human alpha-lactalbumin promoter. Alpha lactalbumin promoter showed significantly higher activity in MDA-MB-435S and T47D breast cancer cells than in normal breast cell lines or other tumor cell lines. We then developed two novel breast cancer-restricted replicative adenoviruses, AdALAE1a and AdE1aALAE1b. In AdALAE1a, expression of adenoviral E1a gene is under the control of alpha-lactalbumin promoter, and in AdE1aALAE1b, expression of both E1a and E1b genes is under the control of a single alpha-lactalbumin promoter. Both breast cancer-restricted replicative adenoviruses showed viral replication efficiency and tumor cell-killing capability similar to wild-type adenovirus in MDA-MB-435S and T47D cells. The replication efficiency and tumor cell-killing capability of both viruses were attenuated significantly in cells that did not support alpha-lactalbumin promoter. AdE1aALAE1b showed better breast cancer restricted replication than AdALAE1a, suggesting that a transcriptional targeting modality with alpha-lactalbumin promoter controlling both E1a and E1b gene expression is superior to alpha-lactalbumin promoter controlling only E1a gene expression. Importantly, we found that AdE1aALAE1b could be used to target hormone-independent breast tumors in vivo by inhibiting the growth of MDA-MB-435S s.c. tumors. These data showed that alpha-lactalbumin promoter could regulate the replication of adenovirus to target hormone-independent breast cancers, suggesting that alpha-lactalbumin promoter can be used to develop a novel therapeutic modality for hormone-independent breast cancer. PMID- 16373701 TI - Homozygous deletions of methylthioadenosine phosphorylase in human biliary tract cancers. AB - The p16(INK4A)/CDKN2A gene on chromosome 9p21 is a site of frequent allelic loss in human cancers, and in a subset of cases, homozygous deletions at this locus encompass the telomeric methylthioadenosine phosphorylase (MTAP) gene. The MTAP gene product is the principal enzyme involved in purine synthesis via the salvage pathway, such that MTAP-negative cancers are solely dependent on de novo purine synthesis mechanisms. Inhibitors of the de novo pathway can then be used to selectively blockade purine synthesis in cancer cells while causing minimal collateral damage to normal cells. In this study, we determine that 10 of 28 (35%) biliary tract cancers show complete lack of Mtap protein expression. In vitro analysis using a selective inhibitor of the de novo purine synthesis pathway, L-alanosine, shows robust growth inhibition in MTAP-negative biliary cancer cell lines CAK-1 and GBD-1 accompanied by striking depletion of intracellular ATP and failure to rescue this depletion via addition of exogenous methylthioadenosine, the principal substrate of the MTAP gene product; in contrast, no significant effects were observed in MTAP-expressing HuCCT1 and SNU308 cell lines. Colony formation studies confirmed that L-alanosine reduced both number and size of CAK-1 colonies in soft agar assays. Knockdown of Mtap protein by RNA interference in L-alanosine-resistant HuCCT1 cells conferred sensitivity to this agent, confirming that intracellular Mtap protein levels determine response to L-alanosine. Inhibitors of de novo purine synthesis can be a potential mechanism-based strategy for treatment of biliary tract cancers, one third of which show complete loss of MTAP function. PMID- 16373702 TI - Transcript profiling in peripheral T-cell lymphoma, not otherwise specified, and diffuse large B-cell lymphoma identifies distinct tumor profile signatures. AB - To glean biological differences and similarities of peripheral T-cell lymphoma not otherwise specified [PTCL-NOS] to diffuse large B-cell lymphoma (DLBCL), a transcriptosome analysis was done on five PTCL-NOS and four DLBCL patients and validated by quantitative real-time reverse transcription-PCR on 10 selected genes. Normal peripheral blood T cells, peripheral blood B cells, and lymph node were used as controls. The resultant gene expression profile delineated distinct "tumor profile signatures" for PTCL-NOS and DLBCL. Several highly overexpressed genes in both PTCL-NOS and DLBCL involve the immune network, stroma, angiogenesis, and cell survival cascades that make important contributions to lymphomagenesis. Inflammatory chemokines and their receptors likely play a central role in these complex interrelated pathways: CCL2 and CXCR4 in PTCL-NOS and CCL5 and CCR1 in DLBCL. Highly overexpressed oncogenes unique to PTCL-NOS are SPI1, STK6, alpha-PDGFR, and SH2D1A, whereas in DLBCL they are PIM1, PIM2, LYN, BCL2A1, and RAB13. Oncogenes common to both lymphomas are MAFB, MET, NF-kappaB2, LCK, and LYN. Several tumor suppressors are also down-regulated (TPTE, MGC154, PTCH, ST5, and SUI1). This study illustrates the relevance of tumor-stroma immune trafficking and identified potential novel prognostic markers and targets for therapeutic intervention. PMID- 16373703 TI - Sensitization of DNA damage-induced apoptosis by the proteasome inhibitor PS-341 is p53 dependent and involves target proteins 14-3-3sigma and survivin. AB - Proteasome inhibition following DNA damage results in the synergistic induction of apoptosis via a nuclear factor-kappaB-independent mechanism. In this study, we identify the role of p53 in mediating apoptosis by the sequence-specific treatment involving the DNA-damaging, topoisomerase I-targeting drug SN-38 followed by the proteasome inhibitor PS-341 (SN-38-->PS-341). The p53-dependent sensitization of DNA damage-induced apoptosis by PS-341 is accompanied by persistent inhibition of proteasome activity and increased cytosolic accumulation of p53, including higher molecular weight forms likely representing ubiquitinated species. In contrast, pretreatment with PS-341 followed by treatment with SN-38 (PS-341-->SN-38), which leads to an antagonistic interaction, results in transient inhibition of proteasome activity and accumulation of significantly lower levels of p53 localized primarily to the nucleus. Whereas cells treated with PS-341-->SN-38 undergo G2 + M cell cycle arrest, cells treated with SN-38- >PS-341 exhibit a decreased G2 + M block with a concomitant increase in the sub G1 population. Decreased accumulation of cells in the G2 + M phase of the cell cycle in SN-38-->PS-341-treated cells compared with PS-341-->SN-38-treated cells correlates with enhanced apoptosis and reduced expression of two p53-modulated proteins, 14-3-3sigma and survivin, both of which play critical roles in regulating G2 + M progression and apoptosis. The functional role of 14-3-3sigma or survivin in regulating the divergent function of p53 in response to SN-38-->PS 341 and PS-341-->SN-38 treatment in inducing apoptosis versus G2 + M arrest/DNA repair, respectively, was confirmed by targeted down-regulation of these proteins. These results provide insights into the mechanisms by which inhibition of proteasome activity modulates DNA damage-induced apoptosis via a p53-dependent pathway. PMID- 16373704 TI - Regulation of signaling phosphoproteins by epidermal growth factor and Iressa (ZD1839) in human endometrial cancer cells that model type I and II tumors. AB - To understand how type I and II endometrial tumors uniquely respond to tyrosine kinase inhibitor treatments, we evaluated the signaling pathways of epidermal growth factor (EGF) receptor (EGFR) under the effects of EGF and Iressa (ZD1839, gefitinib) using Ishikawa H and Hec50co cells that model type I and II endometrial carcinomas, respectively. The cells were assayed for the expression of EGFR and both cell lines express an average of 100,000 EGFR per cell; however, Ishikawa H cells express higher levels of HER-2/neu compared with Hec50co cells (1.38 x 10(5) compared with 2.04 x 10(4), respectively). Using the Kinetworks multi-immunoblotting approach, which profiles 31 signaling phosphoproteins, the most striking result was that Hec50co cells show a higher number of basal phosphorylated sites compared with Ishikawa H cells. Furthermore, we identified targets of Iressa treatment in both cell lines. Iressa, at a dose of 1 micromol/L, blocked the autophosphorylation of EGFR in Ishikawa H and Hec50co cells with some distinctive effects on downstream effectors. Nevertheless, in both cell lines, EGF stimulated and Iressa blocked the major EGFR target mitogen activated protein kinases extracellular signal-regulated kinase 1 and 2 equally. The high basal phosphorylation of numerous signaling molecules in Hec50co cells that were not inhibited by Iressa indicates that other growth factor pathways are active in addition to EGFR. We conclude that endometrial cancer cells that model type I and II carcinomas have the capacity to respond to EGFR inhibition as a therapeutic strategy; however, the response of the more aggressive type II tumors may be limited by the constitutive activation of other signaling pathways. PMID- 16373705 TI - AP23846, a novel and highly potent Src family kinase inhibitor, reduces vascular endothelial growth factor and interleukin-8 expression in human solid tumor cell lines and abrogates downstream angiogenic processes. AB - c-Src is frequently activated in human malignancies, including colon, breast, and pancreatic carcinomas. Several recent studies have shown that activation of Src family kinases leads to tumor progression and metastasis by increasing cellular migration and invasion, promoting cell growth and survival, and deregulating expression of proangiogenic molecules. Therefore, selective inhibitors of Src are being developed for cancer therapy. In this study, we characterize the biological effects of the novel ATP-based Src family kinase inhibitor, AP23846, in tumor cells with high Src activity. As a lead compound, AP23846 is a potent c-Src kinase inhibitor (IC50 approximately 0.5 nmol/L in vitro, approximately 10-fold more potent than PP2, the most widely used commercially available Src family kinase inhibitor). At concentrations of 1 micromol/L, AP23846 led to complete Src inhibition for 48 hours in cells. No cytotoxicity was observed under these conditions, although proliferation rates were slower. Therefore, this was an excellent inhibitor to examine Src-regulated signaling pathways in tumor cells. AP23846 reduced cellular migration, vascular endothelial growth factor, and interleukin-8 in a dose-dependent fashion in pancreatic adenocarcinoma cells grown in vitro. Correspondingly, cell culture supernatants from L3.6pl pancreatic adenocarcinoma cells pretreated with AP23846 failed to promote migration of hepatic endothelial cells in vitro and failed to support angiogenesis into gel foams implanted s.c. in mice in vivo. These results suggest that Src inhibitors affect biological properties of tumor progression and may be useful as cancer therapeutic agents in more advanced disease. PMID- 16373706 TI - Valproic acid induces growth arrest, apoptosis, and senescence in medulloblastomas by increasing histone hyperacetylation and regulating expression of p21Cip1, CDK4, and CMYC. AB - Valproic acid is a well-tolerated anticonvulsant that has been identified recently as a histone deacetylase inhibitor. To evaluate the antitumor efficacy and mechanisms of action of valproic acid in medulloblastoma and supratentorial primitive neuroectodermal tumor (sPNET), which are among the most common malignant brain tumors in children with poor prognosis, two medulloblastoma (DAOY and D283-MED) and one sPNET (PFSK) cell lines were treated with valproic acid and evaluated with a panel of in vitro and in vivo assays. Our results showed that valproic acid, at clinically safe concentrations (0.6 and 1 mmol/L), induced potent growth inhibition, cell cycle arrest, apoptosis, senescence, and differentiation and suppressed colony-forming efficiency and tumorigenicity in a time- and dose-dependent manner. The medulloblastoma cell lines were more responsive than the sPNET cell line and can be induced to irreversible suppression of proliferation and significantly reduced tumorigenicity by 0.6 and 1 mmol/L valproic acid. Daily i.p. injection of valproic acid (400 mg/kg) for 28 days significantly inhibited the in vivo growth of DAOY and D283-MED s.c. xenografts in severe combined immunodeficient mice. With Western hybridization and real-time reverse transcription-PCR, we further showed that the antitumor activities of valproic acid correlated with induction of histone (H3 and H4) hyperacetylation, activation of p21, and suppression of TP53, CDK4, and CMYC expression. In conclusion, valproic acid possesses potent in vitro and in vivo antimedulloblastoma activities that correlated with induction of histone hyperacetylation and regulation of pathways critical for maintaining growth inhibition and cell cycle arrest. Therefore, valproic acid may represent a novel therapeutic option in medulloblastoma treatment. PMID- 16373707 TI - Alterations in the expression of the apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE/Ref-1) in human melanoma and identification of the therapeutic potential of resveratrol as an APE/Ref-1 inhibitor. AB - Apurinic/apyrimidinic endonuclease-1/redox factor-1 (APE/Ref-1) is a multifunctional protein involved in DNA base excision repair and redox regulation of many transcription factors. In different melanoma cell lines, we found that both nucleus and cytoplasm exhibited higher levels of Ref-1 compared with normal melanocytes. Similar increases of Ref-1 expression, detected by immunohistofluorescence, were also evident in nevi and malignant melanoma biopsies compared with normal skin, which were predominantly localized in the nucleus. Using recombinant adenovirus Adref-1, encoding full-length Ref-1, we transiently overexpressed APE/Ref-1 in human melanocytes, which protected these cells from UVB-induced apoptosis and increased foci formation in culture. Ref-1 overexpression also protected melanoma cells from cisplatin- or H2O2-induced apoptosis, whereas increased apoptosis was observed with Ref-1 antisense construct infection. These observations suggested that intracellular Ref-1 levels played an important role in sensitization of melanoma cells to apoptosis. Electrophoretic mobility shift assay results showed that in both cultured primary and metastatic melanomas DNA-binding activities of activator protein-1 and nuclear factor-kappaB were significantly diminished or shifted when anti-APE/Ref 1 antibody was added to deplete APE/Ref-1 from the binding complexes. Induced nuclear factor-kappaB transcriptional activities were also evident after Ref-1 overexpression. Furthermore, using three-dimensional molecular structure modeling and virtual screening, we found that resveratrol, a natural compound found in fruits and vegetables, docks into a druggable pocket of Ref-1 protein. In vitro studies revealed that resveratrol inhibited, in a dose-dependent manner, Ref-1 activated activator protein-1 DNA-binding activities as well as Ref-1 endonuclease activities and rendered melanoma cells more sensitive to dacarbazine treatment. PMID- 16373708 TI - Zinc ribbon domain-containing 1 (ZNRD1) mediates multidrug resistance of leukemia cells through regulation of P-glycoprotein and Bcl-2. AB - Here, we investigated the role of zinc ribbon domain-containing 1 (ZNRD1) in multidrug resistance (MDR) of leukemia cells and the possible underlying mechanisms. ZNRD1 was found overexpressed in the vincristine-induced MDR leukemia cell HL-60/vincristine moreso than its parental cell HL-60. Up-regulation of ZNRD1 expression could confer resistance of both P-glycoprotein (P-gp)-related and P-gp-nonrelated drugs on HL-60 cells and suppress Adriamycin-induced apoptosis accompanied by decreased accumulation and increased releasing amount of Adriamycin. ZNRD1 could significantly up-regulate the expression of P-gp, Bcl-2, and the transcription of the MDR1 gene but not alter the expression of MDR associated protein, glutathione S-transferase activity, or intracellular glutathione content in leukemia cells. In addition, inhibition of ZNRD1 expression by RNA interference or P-gp inhibitor could partially reverse ZNRD1 mediated MDR. The further study of the biological functions of ZNRD1 may be helpful for understanding the mechanisms of MDR of leukemia and developing possible strategies to treat leukemia. PMID- 16373709 TI - Simultaneous targeting of the epidermal growth factor receptor and cyclooxygenase 2 pathways for pancreatic cancer therapy. AB - The aims of this study were to determine the effects of (a) combining the epidermal growth factor receptor (EGFR) blocker (erlotinib) and the cyclooxygenase-2 inhibitor (celecoxib) on cell growth and apoptosis in human pancreatic cancer cell lines, (b) baseline EGFR expression on the potentiation of erlotinib-induced apoptosis by celecoxib, and (c) the effects of the combination on the expression of the COX-2, EGFR, HER-2/neu, and nuclear factor-kappaB (NF kappaB). Baseline expression of EGFR was determined by Western blot analysis in five human pancreatic cancer cell lines. BxPC-3, PANC-1, and HPAC had high EGFR and MIAPaCa had low EGFR. Cells were grown in culture and treated with erlotinib (1 and 10 micromol/L), celecoxib (1 and 10 micromol/L), and the combination. Growth inhibition was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide assay, and apoptosis was assayed by ELISA. Reverse transcriptase-PCR was used to evaluate COX-2 and EGFR mRNA. EGFR, COX-2, and HER 2/neu expression was determined by Western immunoblotting. Electrophoretic mobility shift assay was used to evaluate NF-kappaB activation. Growth inhibition and apoptosis were significantly (P < 0.05) higher in BxPC-3, HPAC, and PANC-1 cells treated with celecoxib and erlotinib than cells treated with either celecoxib or erlotinib. However, no potentiation in growth inhibition or apoptosis was observed in the MIAPaCa cell line with low expression of the EGFR. Significant down-regulation of COX-2 and EGFR expression was observed in the BxPC 3 and HPAC cells treated with the combination of erlotinib (1 micromol/L) and celecoxib (10 micromol/L) compared with celecoxib- or erlotinib-treated cells. Celecoxib significantly down-regulated HER-2/neu expression in BxPC-3 and HPAC cell lines. Significant inhibition of NF-kappaB activation was observed in BxPC-3 and HPAC cell lines treated with erlotinib and celecoxib. (a) Celecoxib can potentiate erlotinib-induced growth inhibition and apoptosis in pancreatic cell lines, (b) high baseline EGFR expression is a predictor of this potentiation, and (c) the down-regulation of EGFR, COX-2, and HER-2/neu expression and NF-kappaB inactivation contributes to the potentiation of erlotinib by celecoxib. PMID- 16373710 TI - Butyric acid prodrugs are histone deacetylase inhibitors that show antineoplastic activity and radiosensitizing capacity in the treatment of malignant gliomas. AB - Histone modification has emerged as a promising approach to cancer therapy. We explored the efficacy of a novel class of histone deacetylase inhibitors in the treatment of malignant gliomas. Treatment of glioma cell lines with two butyric acid derivatives, pivaloylomethyl butyrate (AN-9) and butyroyloxymethyl butyrate (AN-1), induced hyperacetylation, increased p21(Cip1) expression, inhibited proliferation, and enhanced apoptosis. Histone deacetylase inhibitor-induced apoptosis was mediated primarily by caspase-8. Treatment of cells with AN-1 or AN 9 for 24 hours before exposure to gamma-irradiation potentiated further caspase-8 activity and resultant apoptosis. Clonogenic survival curves revealed marked reductions in cell renewal capacity of U251 MG cells exposed to combinations of AN-1 and radiation. Preliminary in vivo experiments using human glioma cell lines grown as xenografts in mouse flanks suggest in vivo efficacy of AN-9. The data suggest that novel butyric acid prodrugs provide a promising treatment strategy for malignant gliomas as single agents and in combination with radiation therapy. PMID- 16373711 TI - Response of endothelial cells to a dual tyrosine kinase receptor inhibition combined with irradiation. AB - Recent studies suggest the possibility of a direct antiangiogenic effect of anti epidermal growth factor receptor (EGFR) drugs due to the presence of EGFR on endothelial cells. The aim of this study was to analyze the direct effect on endothelial cells of associating EGFR targeting, vascular endothelial growth factor receptor (VEGFR)-2 targeting, and irradiation. We examined both the cytotoxic effects and the effect on molecular markers resulting from the combined action of gefitinib (Iressa; anti-EGFR), ZM317450 [VEGFR tyrosine kinase inhibitor (VTKI); anti-VEGFR-2], and irradiation (radiation therapy) on HMME7 cells, an immortalized microvascular endothelial cell of human origin. The presence of a functional EGFR pathway sensitive to gefitinib was shown in HMME7 cells (gefitinib-induced decrease in phospho-EGFR, phospho-p42/p44, and phospho Akt). The stimulation of VEGFR-2 pathway led to an increase in Akt phosphorylation that was inhibited by VTKI. Of note, a post-radiation therapy induction of phospho-p42/p44 was observed on HMME7 cells, and this effect was inhibited by a pretreatment with gefitinib. Based on combination indexes (Chou and Talalay analyses), the associations gefitinib-radiation therapy, VTKI radiation therapy, VTKI-gefitinib, and gefitinib-VTKI-radiation therapy were found to be additive, slightly synergistic, and markedly synergistic, respectively, for the cytotoxicity on HMME7 cells. Among molecular explanatory factors that were examined, the combination gefitinib-radiation therapy totally abolishes DNA-dependent protein kinase expression, and gefitinib attenuates the radiation therapy-induced enhancement of ERCC1 and augments the VTKI-induced CD95 enhancement. The existence of a radiation therapy-dependent neoangiogenesis may be related to the induction of EGFR pathway in endothelial cells, a phenomenon that can be attenuated by anti-EGFR drugs like gefitinib. In complement to the direct antitumor effects of radiation therapy and anti-EGFR drugs, a strong antiangiogenic effect may be obtained with therapeutic strategies combining radiation therapy with EGFR and VEGFR-2 targeting agents. PMID- 16373712 TI - Down-regulation of c-FLIP contributes to the sensitization effect of 3,3' diindolylmethane on TRAIL-induced apoptosis in cancer cells. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor superfamily, which has been shown to preferentially induce apoptosis in cancer cells without adverse effects on normal cells. However, there are still some cancer cells, especially those with high malignancy, resistant to TRAIL-induced apoptosis, impeding the clinical anticancer efficiency of TRAIL. In this report, we showed that 3,3' diindolylmethane, an indole compound derived from cruciferous vegetables, is capable of overcoming TRAIL resistance by sensitizing TRAIL-induced apoptosis in human cancer cells. Noncytotoxic concentrations of 3,3'-diindolylmethane significantly enhanced TRAIL-resistant cancer cells to TRAIL-induced apoptosis via promoting the caspase cascade, a process independent of nuclear factor-kappaB activation and cell surface TRAIL receptor expression. In the search of the molecular mechanisms involved in the sensitization activity of 3,3' diindolylmethane, we found that combined treatment of 3,3'-diindolylmethane and TRAIL led to significant down-regulation of the cellular FLICE inhibitory protein expression (c-FLIP). Furthermore, we provided evidence showing that the reduced c FLIP level is predominately mediated by the ubiquitin-proteasome degradation system. These findings reveal a novel anticancer property of 3,3' diindolylmethane and suggest that this compound could have potential use in cancer therapy to overcome TRAIL resistance. PMID- 16373713 TI - Guggulsterones induce apoptosis and differentiation in acute myeloid leukemia: identification of isomer-specific antileukemic activities of the pregnadienedione structure. AB - In this study, the antileukemic effects of three isomeric pregnadienedione steroids [i.e., cis-guggulsterone, trans-guggulsterone, and 16 dehydroprogesterone] were investigated in HL60 and U937 cells as well as in primary leukemic blasts in culture. Our results show that all three compounds inhibited the proliferation of HL60 and U937 cells, with IC50s ranging from 3.6 to 10.9 micromol/L after treatment for 6 days. These growth inhibitory effects correlated with externalization of phosphatidylserine and loss of mitochondrial membrane potential, suggesting that these isomeric steroids induce apoptosis in leukemia cells. z-VAD-fmk prevented phosphatidylserine externalization but not mitochondrial membrane potential loss, indicating that mitochondrial dysfunction occurred in the absence of caspase activation. Interestingly, although all three compounds increased the generation of reactive oxygen species and decreased phosphorylation of extracellular signal-regulated kinase, only cis-guggulsterone induced a rapid depletion of reduced glutathione levels and oxidation of the mitochondrial phospholipid cardiolipin. 16-Dehydroprogesterone and trans guggulsterone induced differentiation of HL60 and NB4 cells as evidenced by increased surface expression of CD11b and/or CD14, and all three steroids rapidly induced mitochondrial dysfunction and phosphatidylserine externalization of CD34 positive blasts from primary leukemic samples. This study is the first to show that guggulsterones and 16-dehydroprogesterone exert antileukemic effects via the induction of apoptosis and differentiation and, more importantly, identifies the pregnadienedione structure as a potential chemotherapeutic scaffold. PMID- 16373714 TI - In vivo synergy between topoisomerase II and histone deacetylase inhibitors: predictive correlates. AB - Histone deacetylase inhibitors (HDACi) are a promising class of anticancer agents, yet the specific biological effects resulting in cell death are still poorly understood and clinically relevant markers of response are not adequately defined. The anticonvulsant valproic acid has recently emerged as an HDACi, and in vitro studies suggested that valproic acid may potentiate cytotoxic agents. We evaluated the pharmacologic and biological effects of valproic acid on histone acetylation, chromatin structure, and DNA damage induced by topoisomerase II inhibitors in mice bearing breast cancer tumors and developed an ex vivo methodology for response prediction using comet assays. The exposure of mice to valproic acid before exposure to epirubicin led to tumor regression when valproic acid was given for 48 hours at concentrations sufficient for histone hyperacetylation, down-regulation of heterochromatin maintenance proteins, and chromatin decondensation. Tumor response was accurately predicted by ex vivo comet moments. Valproic acid did not exacerbate epirubicin-related toxicity. Antitumor effects were not observed with valproic acid alone despite biologically active valproic acid concentrations. These findings suggest that exposure of tumor-bearing mice to valproic acid potentiated the antitumor effects of topoisomerase II inhibitors without enhancing toxicity. The HDACi-induced histone acetylation and modulation of heterochromatin correlated with potentiation of epirubicin-mediated DNA damage. However, these effects did not result in antitumor activity when using a HDACi alone and hence should not be considered a surrogate marker. Ex vivo comet assays may be useful as a predictive tool when tumor cells are limited and serial biopsies are difficult to obtain. PMID- 16373715 TI - Class III beta-tubulin expression in tumor cells predicts response and outcome in patients with non-small cell lung cancer receiving paclitaxel. AB - Both fundamental and clinical studies suggest that class III beta-tubulin expression is associated with resistance to taxanes and constitutes a prognostic factor in several solid tumors. In this study, we assessed the prognostic and predictive value of class III beta-tubulin in tumors of patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) treated with paclitaxel based or other regimens that did not include tubulin-binding agents. Expression of class III beta-tubulin was examined immunohistochemically in 91 tumor samples obtained before treatment from patients with stage III and IV NSCLC, including 47 who received paclitaxel-based regimens and 44 who received regimens without tubulin-binding agents. Response to chemotherapy, progression-free survival, and overall survival were correlated with the expression of class III beta-tubulin protein. The response rate was 37.5% (16 responses among 45 evaluable patients) among patients receiving paclitaxel. Patients whose tumors expressed low levels of class III beta-tubulin isotype had a better response rate, longer progression free survival, and overall survival (P < 0.001, 0.004, and 0.002, respectively), whereas this variable was not found to be predictive in patients receiving regimens without tubulin-binding agents. A multivariate analysis taking into account sex, age, histology, stage, and class III beta-tubulin confirmed that low level class III beta-tubulin expression was independently correlated with progression-free survival (P = 0.003) and overall survival (P = 0.003). These findings suggest that the expression levels of class III beta-tubulin in tumor cells is predictive of response to therapy and patient outcome in patients with NSCLC receiving paclitaxel-based chemotherapy but is not a general prognostic factor in this patient population. PMID- 16373716 TI - Imatinib binding and cKIT inhibition is abrogated by the cKIT kinase domain I missense mutation Val654Ala. AB - Several activating mutations in the cKIT receptor tyrosine kinase are associated with the development and progression of gastrointestinal stromal tumors (GIST). Treatment of GIST with the tyrosine kinase inhibitor imatinib (Gleevec, STI571; Novartis, Basel, Switzerland) increases patient survival. However, many patients develop resistance to imatinib following initial responses. We sequenced cKIT exons from two patients with GIST after the development of imatinib resistance, revealing a point mutation in kinase domain I (exon 13), Val654Ala, which has been associated previously with relapse and resistance. Molecular modeling of cKIT-imatinib complexes shows that this residue is located in the drug-binding site and that the Val654Ala mutation disrupts drug binding by removing hydrophobic contacts with the central diaminophenyl ring of imatinib. Loss of these contacts results in a destabilizing effect on two key hydrogen bonds between imatinib and Asp310 and Thr670 of cKIT. Calculations based on published crystallography data show an estimated destabilization energy of 2.25 kcal/mol in the Val654Ala cKIT compared with wild type. When present on the same cKIT allele as an oncogenic mutation, the Val654Ala mutation abolishes imatinib-mediated inhibition of cKIT phosphoactivation in vitro. These results highlight some of the structural and functional consequences of the Val654Ala mutation in relapsing imatinib-resistant GIST and emphasize the importance of tumor genetics in drug development and patient-specific cancer treatment regimens. PMID- 16373717 TI - Cross-talk between DNA damage and cell survival checkpoints during G2 and mitosis: pharmacologic implications. AB - In this study, we wanted to clarify the role of survivin-mediated survival signaling during G2 and M in tumor cells treated with DNA-damaging agents. As a cellular model, we selected MOLT-4 human T-cell lymphoblastic leukemia cells that overexpress survivin and nonfunctional p53. Treatment with melphalan, a classic DNA-damaging agent, led to the induction of the DNA damage checkpoint and growth arrest in the G2 phase of the cell cycle. Checkpoint abrogation by caffeine was accompanied by mitotic entry and rapid apoptotic cell death, whereas cells remaining in G2 remained viable during the same time interval. Unexpectedly, when the spindle checkpoint was activated following G2 abrogation, two different effects could be observed. If the microtubules of the melphalan-treated cells were destabilized by nocodazole, cells became arrested in prometaphase with low survivin levels and entered apoptosis. In contrast, if the microtubules of the melphalan-treated cells were stabilized by taxol, cells were still arrested in prometaphase, but apoptotic execution was inhibited. This effect is, most likely, directly mediated by survivin itself given its well-established antiapoptotic functions. In conclusion, depending on the way the spindle checkpoint was activated in cells with damaged DNA, cells could be either protected by survivin or die during mitosis. We suggest that the efficacy of DNA damage checkpoint abrogators used in combination with DNA-damaging agents may critically depend on whether DNA damage is able to invoke spindle checkpoint response and to activate survivin-associated survival signaling during mitosis. PMID- 16373718 TI - Chemotherapy and TRAIL-mediated colon cancer cell death: the roles of p53, TRAIL receptors, and c-FLIP. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has recently attracted attention as a potential therapeutic agent in the treatment of cancer. We assessed the roles of p53, TRAIL receptors, and cellular Fas-associated death domain-like interleukin-1beta-converting enzyme inhibitory protein (c-FLIP) in regulating the cytotoxic effects of recombinant TRAIL (rTRAIL) alone and in combination with chemotherapy [5-fluorouracil (5-FU), oxaliplatin, and irinotecan] in a panel of colon cancer cell lines. Using clonogenic survival and flow cytometric analyses, we showed that chemotherapy sensitized p53 wild-type, mutant, and null cell lines to TRAIL-mediated apoptosis. Although chemotherapy treatment did not modulate mRNA or cell surface expression of the TRAIL receptors death receptor 4, death receptor 5, decoy receptor 1, or decoy receptor 2, it was found to down-regulate expression of the caspase-8 inhibitor, c-FLIP. Stable overexpression of the long c-FLIP splice form but not the short form was found to inhibit chemotherapy/rTRAIL-induced apoptosis. Furthermore, siRNA-mediated down regulation of c-FLIP, particularly the long form, was found to sensitize colon cancer cells to rTRAIL-induced apoptosis. In addition, treatment of a 5-FU resistant cell line with 5-FU down-regulated c-FLIP expression and sensitized the chemotherapy-resistant cell line to rTRAIL. We conclude that TRAIL-targeted therapies may be used to enhance conventional chemotherapy regimens in colon cancer regardless of tumor p53 status. Furthermore, inhibition of c-FLIP may be a vital accessory strategy for the optimal use of TRAIL-targeted therapies. PMID- 16373719 TI - Anti-CD70 antibodies: a potential treatment for EBV+ CD70-expressing lymphomas. AB - A monoclonal antibody (Rituximab) directed against the B-cell surface antigen, CD20, is increasingly used as a therapy for B-cell lymphomas. However, CD20 is expressed on all normal mature B cells and hence is not a specific tumor target. In contrast, CD70 is expressed on highly activated lymphocytes as well as on many B-cell and T-cell lymphomas but is not expressed on the great majority of B cells and T cells. In this report, we have explored the potential utility of anti-CD70 monoclonal antibodies for treatment of CD70+ EBV+ B-cell lymphomas. Using two Burkitt's lymphoma lines (Raji and Jijoye) that express surface CD70 and a CD70- Burkitt's lymphoma line (Akata), we show that two different monoclonal antibodies directed against human CD70 allow rabbit and human complement to kill EBV+ B cells in a CD70-dependent manner in vitro. In the absence of complement, neither anti-CD70 antibody induced in vitro killing of CD70+ cell lines. Importantly, i.p. injection of anti-CD70 antibodies also inhibited the growth of CD70+ Burkitt's lymphoma cells in severe combined immunodeficient mice but did not inhibit the growth of CD70- Burkitt's lymphoma cells. These results suggest that anti-CD70 antibodies may be useful for the treatment of CD70+ B-cell lymphomas. PMID- 16373720 TI - Sharing evidence on humanitarian relief. PMID- 16373721 TI - How Islam changed medicine. PMID- 16373722 TI - How Google is changing medicine. PMID- 16373723 TI - Choice. PMID- 16373724 TI - Happiness. PMID- 16373725 TI - White coats and fingerprints: diagnostic reasoning in medicine and investigative methods of fictional detectives. PMID- 16373726 TI - Doctors, detectives, and common sense. PMID- 16373727 TI - The case of the disappearing teaspoons: longitudinal cohort study of the displacement of teaspoons in an Australian research institute. AB - OBJECTIVES: To determine the overall rate of loss of workplace teaspoons and whether attrition and displacement are correlated with the relative value of the teaspoons or type of tearoom. DESIGN: Longitudinal cohort study. SETTING: Research institute employing about 140 people. SUBJECTS: 70 discreetly numbered teaspoons placed in tearooms around the institute and observed weekly over five months. MAIN OUTCOME MEASURES: Incidence of teaspoon loss per 100 teaspoon years and teaspoon half life. RESULTS: 56 (80%) of the 70 teaspoons disappeared during the study. The half life of the teaspoons was 81 days. The half life of teaspoons in communal tearooms (42 days) was significantly shorter than for those in rooms associated with particular research groups (77 days). The rate of loss was not influenced by the teaspoons' value. The incidence of teaspoon loss over the period of observation was 360.62 per 100 teaspoon years. At this rate, an estimated 250 teaspoons would need to be purchased annually to maintain a practical institute-wide population of 70 teaspoons. CONCLUSIONS: The loss of workplace teaspoons was rapid, showing that their availability, and hence office culture in general, is constantly threatened. PMID- 16373728 TI - Stains on the carpet. PMID- 16373729 TI - The emergence of ringing vertigo. PMID- 16373730 TI - Barbering in mice: a model for trichotillomania. PMID- 16373731 TI - Harry Potter casts a spell on accident prone children. PMID- 16373732 TI - Revisiting the lot of the first incestuous family: the biblical origins of shifting the blame on to female family members. PMID- 16373733 TI - Sex symbols ancient and modern: their origins and iconography on the pedigree. PMID- 16373734 TI - Seen from the other side: visual experiences during cataract surgery under topical anaesthesia. PMID- 16373735 TI - Shape of glass and amount of alcohol poured: comparative study of effect of practice and concentration. AB - OBJECTIVE: To determine whether people pour different amounts into short, wide glasses than into tall, slender ones. DESIGN: College students practised pouring alcohol into a standard glass before pouring into larger glasses; bartenders poured alcohol for four mixed drinks either with no instructions or after being told to take their time. SETTING: University town and large city, United States. PARTICIPANTS: 198 college students and 86 bartenders. MAIN OUTCOME MEASURES: Volume of alcohol poured into short, wide and tall, slender glasses. RESULTS: Aiming to pour a "shot" of alcohol (1.5 ounces, 44.3 ml), both students and bartenders poured more into short, wide glasses than into tall slender glasses (46.1 ml v 44.7 ml and 54.6 ml v 46.4 ml, respectively). Practice reduced the tendency to overpour, but not for short, wide glasses. Despite an average of six years of experience, bartenders poured 20.5% more into short, wide glasses than tall, slender ones; paying careful attention reduced but did not eliminate the effect. CONCLUSIONS: To avoid overpouring, use tall, narrow glasses or ones on which the alcohol level is premarked. To avoid underestimating the amount of alcohol consumed, studies using self reports of standard drinks should ask about the shape of the glass. PMID- 16373736 TI - Interventions for preventing or treating alcohol hangover: systematic review of randomised controlled trials. AB - OBJECTIVE: To assess the clinical evidence on the effectiveness of any medical intervention for preventing or treating alcohol hangover. DATA SOURCES: Systematic searches on Medline, Embase, Amed, Cochrane Central, the National Research Register (UK), and ClincalTrials.gov (USA); hand searches of conference proceedings and bibliographies; contact with experts and manufacturers of commercial preparations. Language of publication was not restricted. STUDY SELECTION AND DATA EXTRACTION: All randomised controlled trials of any medical intervention for preventing or treating alcohol hangover were included. Trials were considered if they were placebo controlled or controlled against a comparator intervention. Titles and abstracts of identified articles were read and hard copies were obtained. The selection of studies, data extraction, and validation were done independently by two reviewers. The Jadad score was used to evaluate methodological quality. RESULTS: Fifteen potentially relevant trials were identified. Seven publications failed to meet all inclusion criteria. Eight randomised controlled trials assessing eight different interventions were reviewed. The agents tested were propranolol, tropisetron, tolfenamic acid, fructose or glucose, and the dietary supplements Borago officinalis (borage), Cynara scolymus (artichoke), Opuntia ficus-indica (prickly pear), and a yeast based preparation. All studies were double blind. Significant intergroup differences for overall symptom scores and individual symptoms were reported only for tolfenamic acid, gamma linolenic acid from B officinalis, and a yeast based preparation. CONCLUSION: No compelling evidence exists to suggest that any conventional or complementary intervention is effective for preventing or treating alcohol hangover. The most effective way to avoid the symptoms of alcohol induced hangover is to practise abstinence or moderation. PMID- 16373737 TI - Depictions of substance use in reality television: a content analysis of The Osbournes. AB - OBJECTIVE: To determine the source and slant of messages in a reality television programme that may promote or inhibit health related or risky behaviours. DESIGN: Coding visual and verbal references to alcohol, tobacco, and other drug (ATOD) use in The Osbournes. REVIEW METHODS: Three reviewers watched all 10 episodes of the first season and coded incidents of substance use according to the substance used (alcohol, tobacco, or drugs), the way use was portrayed (visually or verbally), the source of the message (the character in the show involved in the incident), and the slant of the incident (endorsement or rejection). MAIN OUTCOME MEASURES: The variation in number of messages in an average episode, the slant of messages, and message source. RESULTS: The average number of messages per episode was 9.1 (range 2-17). Most drug use messages (15, 54%) implied rejection of drugs, but most alcohol messages (30, 64%) and tobacco messages (12, 75%) implied endorsements for using these substances. Most rejections (34, 94%) were conveyed verbally, but most endorsements (36, 65%) were conveyed visually. Messages varied in frequency and slant by source. CONCLUSIONS: The reality television show analysed in this study contains numerous messages on substance use that imply both rejection and endorsement of use. The juxtaposition of verbal rejection messages and visual endorsement messages, and the depiction of contradictory messages about substance use from show characters, may send mixed messages to viewers about substance use. PMID- 16373738 TI - The wealth of distinguished doctors: retrospective survey. AB - OBJECTIVE: To assess changes in the wealth of distinguished doctors in the United Kingdom between 1860 and 2001. DESIGN: Retrospective survey. SETTING: The UK. PARTICIPANTS: 980 doctors of sufficient distinction to be included in the Oxford Dictionary of National Biography and who died between 1860 and 2001. MAIN OUTCOME MEASURES: Wealth at death, based on probate records and adjusted relative to average earnings in 2002. RESULTS: The wealth of distinguished doctors declined substantially between 1860 and 2001, and paralleled a decline in the relative income of doctors in general. The wealth of distinguished doctors also declined relative to other groups of distinguished individuals. CONCLUSIONS: In the 19th century, distinction in doctors was accompanied by substantial wealth, whereas by the end of the 20th century, the most distinguished doctors were less wealthy than their contemporaries who had achieved national distinction in other areas. PMID- 16373739 TI - Judging a book by its cover: descriptive survey of patients' preferences for doctors' appearance and mode of address. AB - OBJECTIVE: To document patients' preferred dress styles of their doctors and modes of address. DESIGN: Descriptive survey. SETTING: Inpatients and outpatients at a tertiary level hospital, New Zealand. PARTICIPANTS: 202 inpatients and 249 outpatients, mean age 55.9 (SD 19.3) years. MAIN OUTCOME MEASURES: Ranking of patients' opinions of photographs showing doctors wearing different dress styles. A five point Likert scale was used to measure patient comfort with particular items of appearance. RESULTS: Patients preferred doctors to wear semiformal attire, but the addition of a smiling face was even better. The next most preferred styles were semiformal without a smile, followed by white coat, formal suit, jeans, and casual dress. Patients were more comfortable with conservative items of clothing, such as long sleeves, covered shoes, and dress trousers or skirts than with less conservative items such as facial piercing, short tops, and earrings on men. Many less conservative items such as jeans were still acceptable to most patients. Most patients preferred to be called by their first name, to be introduced to a doctor by full name and title, and to see the doctor's name badge worn at the breast pocket. Older patients had more conservative preferences. CONCLUSIONS: Patients prefer doctors to wear semiformal dress and are most comfortable with conservative items; many less conservative items were, however, acceptable. A smile made a big difference. PMID- 16373740 TI - Patients bearing gifts: are there strings attached? PMID- 16373741 TI - Beatty's box. PMID- 16373742 TI - Painting the history of cardiology. PMID- 16373743 TI - Influenza, 1918. PMID- 16373744 TI - Epidemiology and prognosis of coma in daytime television dramas. AB - OBJECTIVE: To determine how soap operas portray, and possibly misrepresent, the likelihood of recovery for patients in coma. DESIGN: Retrospective cohort study. SETTING: Nine soap operas in the United States reviewed between 1 January 1995 and 15 May 2005. SUBJECTS: 64 characters who experienced a period of unconsciousness lasting at least 24 hours. Their final status at the end of the follow-up period was compared with pooled data from a meta-analysis. RESULTS: Comas lasted a median of 13 days (interquartile range 7-25 days). Fifty seven (89%) patients recovered fully, five (8%) died, and two (3%) remained in a vegetative state. Mortality for non-traumatic and traumatic coma was significantly lower than would be predicted from the meta-analysis data (non traumatic 4% v 53%; traumatic 6% v 67%; Fisher's exact test both P < 0.001). On the day that patients regained consciousness, most (49/57; 86%) had no evidence of limited function, cognitive deficit, or residual disability needing rehabilitation. Compared with meta-analysis data, patients in this sample had a much better than expected chance of returning to normal function (non-traumatic 91% v 1%; traumatic 89% v 7%; both P < 0.001). CONCLUSIONS: The portrayal of coma in soap operas is overly optimistic. Although these programmes are presented as fiction, they may contribute to unrealistic expectations of recovery. PMID- 16373745 TI - From Shakespeare to Star Trek and beyond: a Medline search for literary and other allusions in biomedical titles. AB - OBJECTIVES: To document biomedical paper titles containing literary and other allusions. DESIGN: Retrospective survey. SETTING: Medline (1951 to mid-2005) through Dialog Datastar. MAIN OUTCOME MEASURE: Allusions to Shakespeare, Hans Christian Andersen, proverbs, the Bible, Lewis Carroll, and movie titles, corrected and scaled for five year periods 1950-4 to 2000-4. RESULTS: More than 1400 Shakespearean allusions exist, a third of them to "What's in a name" and another third to Hamlet-mostly to "To be or not to be." The trend of increasing use of allusive titles, identified from Shakespeare and Andersen, is paralleled by allusions to Carroll and proverbs; the trend of biblical allusions is also upward but is more erratic. Trends for newer allusions are also upwards, including the previously surveyed "paradigm shift." Allusive titles are likely to be to editorial or comment rather than to original research. CONCLUSIONS: The similar trends are presumably a mark of a particular learnt author behaviour. Newer allusions may be becoming more popular than older ones. Allusive titles can be unhelpful to reviewers and researchers, and many are now cliches. Whether they attract readers or citations is unknown, but better ways of gaining attention exist. PMID- 16373746 TI - Cinderella revisited. AB - OBJECTIVE: To examine the references to Cinderella in medical literature. DESIGN: Analysis of papers published in the past 50 years that mention Cinderella. RESULTS: The trend for use of Cinderella as a metaphor in medical publications is increasing exponentially. Five separate themes emerged: neglect, identity, transformation, exhaustion, and the mixed metaphor. CONCLUSIONS: The medical use of the Cinderella fable is growing in popularity. PMID- 16373747 TI - Obesity: the elephant in the corner. PMID- 16373748 TI - What is successful ageing and who should define it? PMID- 16373749 TI - Slow tracking for BMJ papers. PMID- 16373750 TI - Science to practice: can fluoroscopic radiation dose be substantially reduced? PMID- 16373756 TI - Quality assurance: the missing link. PMID- 16373757 TI - Evolving and experimental technologies in medical imaging. AB - Medical images are created by detecting radiation probes transmitted through or emitted or scattered by the body. The radiation, modulated through interactions with tissues, yields patterns that provide anatomic and/or physiologic information. X-rays, gamma rays, radiofrequency signals, and ultrasound waves are the standard probes, but others like visible and infrared light, microwaves, terahertz rays, and intrinsic and applied electric and magnetic fields are being explored. Some of the younger technologies, such as molecular imaging, may enhance existing imaging modalities; however, they also, in combination with nanotechnology, biotechnology, bioinformatics, and new forms of computational hardware and software, may well lead to novel approaches to clinical imaging. This review provides a brief overview of the current state of image-based diagnostic medicine and offers comments on the directions in which some of its subfields may be heading. PMID- 16373758 TI - Diagnostic architectural and dynamic features at breast MR imaging: multicenter study. AB - PURPOSE: To prospectively determine the prevalence and predictive value of three dimensional (3D) and dynamic breast magnetic resonance (MR) imaging and contrast material kinetic features alone and as part of predictive diagnostic models. MATERIALS AND METHODS: The study protocol was approved by the institutional review board or ethics committees of all participating institutions, and informed consent was obtained from all participants. Although study data collection was performed before HIPAA went into effect, standards that would be compliant with HIPAA were adhered to. Data from the International Breast MR Consortium trial 6883 were used in the analysis. Women underwent 3D (minimum spatial resolution, 0.7 x 1.4 x 3 mm; minimal temporal resolution, 4 minutes) and dynamic two dimensional (temporal resolution, 15 seconds) MR imaging examinations. Readers rated enhancement shape, enhancement distribution, border architecture, enhancement intensity, presence of rim enhancement or internal septations, and the shape of the contrast material kinetic curve. Regression was performed for each feature individually and after adjustment for associated mammographic findings. Multivariate models were also constructed from multiple architectural and dynamic features. Areas under the receiver operating characteristic curve (Az values) were estimated for all models. RESULTS: There were 995 lesions in 854 women (mean age, 53 years +/- 12 [standard deviation]; range, 18-80 years) for whom pathology data were available. The absence of enhancement was associated with an 88% negative predictive value for cancer. Qualitative characterization of the dynamic enhancement pattern was associated with an Az value of 0.66 across all lesion architectures. Focal mass margins (Az = 0.76) and signal intensity (Az = 0.70) were highly predictive imaging features. Multivariate models were constructed with an Az value of 0.880. CONCLUSION: Architectural and dynamic features are important in breast MR imaging interpretation. Multivariate models involving feature assessment have a diagnostic accuracy superior to that of qualitative characterization of the dynamic enhancement pattern. PMID- 16373759 TI - High-performance wavelet compression for mammography: localization response operating characteristic evaluation. AB - PURPOSE: To evaluate the accuracy of a visually lossless, image-adaptive, wavelet based compression method for achievement of high compression rates at mammography. MATERIALS AND METHODS: The study was approved by the institutional review board of the University of South Florida as a research study with existing medical records and was exempt from individual patient consent requirements. Patient identifiers were obliterated from all images. The study was HIPAA compliant. An algorithm based on scale-specific quantization of biorthogonal wavelet coefficients was developed for the compression of digitized mammograms with high spatial and dynamic resolution. The method was applied to 500 normal and abnormal mammograms from 278 patients who were 32-85 years old, 85 of whom had biopsy-proved cancer. Film images were digitized with a charge-coupled device based digitizer. The original and compressed reconstructed images were evaluated in a localization response operating characteristic experiment involving three radiologists with 2-10 years of experience in reading mammograms. RESULTS: Compression rates in the range of 14:1 to 2051:1 were achieved, and the rates were dependent on the degree of parenchymal density and the type of breast structure. Ranges of the area under the receiver operating characteristic curve were 0.70-0.83 and 0.72-0.86 for original and compressed reconstructed mammograms, respectively. Ranges of the area under the localization response operating characteristic curve were 0.39-0.65 and 0.43-0.71 for original and compressed reconstructed mammograms, respectively. The localization accuracy increased an average of 6% (0.04 of 0.67) with the compressed mammograms. Localization performance differences were statistically significant with P = .05 and favored interpretation with the wavelet-compressed reconstructed images. CONCLUSION: The tested wavelet-based compression method proved to be an accurate approach for digitized mammography and yielded visually lossless high-rate compression and improved tumor localization. PMID- 16373760 TI - Multi-detector row CT coronary angiography: influence of reconstruction technique and heart rate on image quality. AB - PURPOSE: To prospectively evaluate to what extent image quality in 16-detector row computed tomographic (CT) coronary angiography is a function of the heart rate and the image reconstruction technique used. MATERIALS AND METHODS: A total of 70 patients (49 men, 21 women; mean age, 59.1 years +/- 5.8 [standard deviation]) consecutively underwent multi-detector row CT coronary angiography; 49 patients additionally underwent coronary angiography. Image reconstruction was based on both relative and absolute timing. A total of 20 equidistant relative and absolute image reconstructed intervals were assessed by applying a four-step grading scale. Cluster and discrimination analysis, Spearman correlation analysis, and Wilcoxon and chi2 tests were used for statistical analysis. Institutional review board approval and written informed consent were obtained. RESULTS: Though significantly (P < .001) better image quality was observed for image reconstruction based on absolute timing and in patients with lower heart rates, influence on diagnostic accuracy was not significant. Irrespective of the reconstruction technique used, best image quality was observed in patients with a low heart rate for middiastolic reconstruction intervals (starting points: 61% of R-R interval [range, 40%-75%] and 599.3 msec after R [range, 450-840 msec]) and in patients with a high heart rate for end-systolic or early-diastolic intervals (starting points: 27.3% of R-R interval [range, 10%-45%] and 202.3 msec after R [range, 82-336 msec]). With regard to the vessel section and reconstruction technique, cutoff heart rates of the intervals were 64.0-68.5 beats per minute. Patients with stenoses of more than 50% were identified with 86% sensitivity and specificity, and there was no significant difference between relative and absolute timing (P = .99). CONCLUSION: In multi-detector row CT coronary angiography, image quality depends on the choice of a suited reconstruction interval. In patients with high heart rates, the best image quality can be obtained with end-systolic and early-diastolic intervals; in patients with low heart rates, the best results are achieved with middiastolic intervals. PMID- 16373761 TI - Radiation exposure reduction during voiding cystourethrography in a pediatric porcine model of vesicoureteral reflux. AB - PURPOSE: To compare grid-controlled variable-rate pulsed fluoroscopy (GCPFL) and continuous fluoroscopy (CFL) for the reduction of radiation exposure during voiding cystourethrography (VCUG) in a pediatric porcine model of vesicoureteral reflux. MATERIALS AND METHODS: Institutional animal care and use committee approval was obtained. Vesicoureteral reflux was simulated in four pigs, and 48 VCUG studies were performed (24 with GCPFL, 24 with CFL). VCUG was performed at abdominal girths of 8-10 cm (group 1, simulates human newborn to 6-month-old infant), 12-13 cm (group 2, simulates 2-3-year-old child), and 15-17 cm (group 3, simulates 10-year-old child). An electronic device calculated total radiation exposure during fluoroscopy and image recording. With five-point ordinal scales, VCUG images were scored independently for anatomic conspicuity and overall diagnostic quality by two radiologists (radiologists A and B). An analysis of variance was used to compare radiation exposures and fluoroscopy times between GCPFL and CFL and to determine whether radiation exposure and fluoroscopy time were dependent on the pig's abdominal girth. The Pearson product-moment correlation coefficient was used to assess whether fluoroscopy time was correlated with radiation exposure. Anatomic conspicuity and diagnostic quality scores were compared by means of the Wilcoxon signed rank test. RESULTS: Results of analysis of variance revealed that GCPFL resulted in a significant reduction in total radiation exposure compared with CFL for each of the three groups (P < .05 for each comparison), and this reduction was most marked in the larger animals. There were no significant differences in diagnostic quality of the recorded VCUG images (P > .05). Anatomic conspicuity was not significantly different for groups 2 and 3, but there was a significantly higher score for GCPFL in group 1 for radiologist A (P = .04). CONCLUSION: By using GCPFL in the performance of VCUG in a pediatric porcine model of vesicoureteral reflux, total radiation exposure can be reduced by a factor of 4.6-7.5 lower than with CFL, and diagnostic-quality images can be obtained. PMID- 16373762 TI - Percutaneous MR imaging-guided transvascular access of mesenteric venous system: study in swine model. AB - PURPOSE: To determine if, with use of magnetic resonance (MR) imaging guidance alone, transcaval puncture of the superior mesenteric vein (SMV) and/or portal vein is feasible with a percutaneous femoral vein approach. MATERIALS AND METHODS: The Institutional Animal Care and Use Committee approved the animal studies. Ten inferior vena cava (IVC)-SMV punctures were performed in six pigs. An active MR intravascular needle system was used for all transvascular punctures, and all procedures were performed with a 1.5-T MR unit. The needle was introduced via a 12-F femoral vein sheath and advanced into the IVC by using a real-time gradient-recalled-echo sequence (3.4/1.2 [repetition time msec/echo time msec], 45 degrees flip angle, and six to eight frames per second). Fast transverse spoiled gradient-recalled acquisition in the steady state (SPGR) (6.0/1.5, 60 degrees flip angle, one frame per second) was performed to confirm needle trajectory. The needle system was advanced under real-time MR imaging to puncture the SMV. The location of the needle tip was confirmed with a fast spin echo sequence (1904/4.5, 36-cm field of view). A direct MR portogram was obtained after the administration of gadopentetate dimeglumine at a concentration of 25% with fast SPGR (6/1.3, 90 degrees flip angle, no section selection, three frames per second). Success was defined as entry into the mesenteric venous system without traversal of any retroperitoneal organs or adjacent vasculature. RESULTS: Successful MR imaging-guided IVC-SMV punctures were performed in all 10 procedures (100%). The needle was fully visualized as it traversed the retroperitoneum and entered the SMV. MR portograms were successfully obtained following all punctures through the needle. Conventional transverse MR imaging helped confirm that the needle did not traverse any retroperitoneal organs or vessels. CONCLUSION: With use of only MR imaging guidance and an active MR imaging intravascular needle system, the authors were able to successfully puncture the SMV from the IVC with direct visualization of the needle and all retroperitoneal structures. PMID- 16373763 TI - Roux-en-Y gastric bypass surgery for morbid obesity: evaluation of postoperative extraluminal leaks with upper gastrointestinal series. AB - PURPOSE: To retrospectively evaluate the radiographic features of extraluminal leak after Roux-en-Y gastric bypass (RYGBP) surgery at upper gastrointestinal (GI) examinations in a large series of patients and to determine morbidity and mortality in those patients with leak. MATERIALS AND METHODS: The investigational review board approved this HIPAA-compliant study, and the need for patient informed consent was waived. Radiologic database review revealed 1202 upper GI studies performed over a 4-year period in 906 patients after RYGBP. Extraluminal leak was identified in 50 patients. Two patients with leaks that occurred before the study period were excluded. Of the remaining 48 patients, 12 were men and 36 were women (mean age, 45 years; range, 26-64 years). Surgery had been laparoscopic in 23 patients and open in 25. Upper GI studies were analyzed by two radiologists in consensus for the origin, extent, and severity of leaks and associated findings. Chart review was performed to determine clinical course, treatment, and outcome. RESULTS: Fifty extraluminal leaks were detected in 48 of 904 patients (5.3%) at upper GI examinations. All leaks were identified within 28 days, and, in 37 of 48 patients (77%), leakage was diagnosed within 1 week of surgery. The majority of leaks (n = 37) originated from the gastrojejunal anastomosis. Leaks also occurred at the distal portion of the esophagus (n = 5), the gastric pouch (n = 5), the oversewn jejunum (n = 2), and the distal anastomosis (n = 1). Leaks extended into the left upper quadrant in 30 patients. Obstruction or ileus was present in 35 of 48 patients (73%). Leak into the excluded stomach was observed in 15 of 48 patients. The occurrence of extraluminal leak prolonged hospital stays; organ failure occurred in 14 (29%) and death in three (6%) of the 48 patients. CONCLUSION: Extraluminal leak was identified on upper GI series in 48 of 904 patients (5.3%) after RYGBP for morbid obesity. Extraluminal leak most commonly arises from the gastrojejunal anastomosis and extends into the left upper quadrant. Extraluminal leak affects morbidity and mortality. PMID- 16373764 TI - Crohn disease of the small bowel: preliminary comparison among CT enterography, capsule endoscopy, small-bowel follow-through, and ileoscopy. AB - PURPOSE: To prospectively compare four diagnostic small-bowel imaging techniques for depiction of abnormal findings in the same patients known to have or suspected of having Crohn disease. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. Patients known to have or suspected of having nonobstructive Crohn disease were recruited. Each patient underwent capsule endoscopy, computed tomographic (CT) enterography, colonoscopy with ileoscopy, and small-bowel follow-through (SBFT). Findings consistent with Crohn disease were tabulated for each imaging examination (diagnostic yield). The proportions of patients with positive findings at each examination were compared, and any significant differences between the tests were calculated by using the exact McNemar test. RESULTS: Seventeen patients (nine women, eight men; mean age, 49 years; range, 18-78 years) completed the study out of 20 patients enrolled. Crohn disease was depicted by capsule endoscopy in 12 patients (71%), ileoscopy in 11 (65%), CT enterography in nine (53%), and SBFT in four (24%). Ileoscopy was incomplete in four patients, and capsule endoscopy was incomplete in two patients. Capsule endoscopy had the highest diagnostic yield for Crohn disease, and SBFT had the lowest, but these differences were not statistically significant (P = .02). SBFT failed to depict a stricture in one patient, which resulted in surgical removal of the capsule. CT enterography and SBFT depicted extraintestinal findings (eg, mesenteric adenopathy in two patients, perianal and enterocolic fistulas in one patient) not detected endoscopically. CONCLUSION: This preliminary study demonstrates capsule endoscopy and CT enterography may depict nonobstructive Crohn disease when techniques such as ileoscopy and SBFT have negative or inconclusive findings. PMID- 16373765 TI - Types and frequencies of biliary tract variations associated with a major portal venous anomaly: analysis with multi-detector row CT cholangiography. AB - PURPOSE: To retrospectively determine whether major portal venous variation was more frequently associated with biliary variants, with consideration of the types and frequencies of biliary tract variations in the right and left liver lobes. MATERIALS AND METHODS: Before undergoing computed tomographic (CT) cholangiography, patients gave informed consent. The retrospective research protocol was approved, and informed consent was waived by the ethics committee. Forty-four patients aged 29-80 years who underwent multi-detector row CT cholangiography had a major portal vein variation in which the main portal vein diverged into the common trunk of the left portal vein and right anterior sectorial portal vein. One hundred fifty-eight consecutive patients aged 26-89 years who did not have this variation served as the control group. Three radiologists retrospectively evaluated the confluence pattern of the bile duct, the relationship between this pattern and the portal vein, and the major branching pattern of the portal vein. Pearson chi2 and Fisher exact tests were performed to identify significant differences between the two patient groups. RESULTS: The classic hilar confluence pattern, where the right posterior sectorial duct connects supraportally with the right anterior sectorial duct, was less frequent in the patients with the portal vein variation than in the control subjects (32% vs 73%, P < .05). The following biliary tract variations were identified more frequently in the variation group than in the control group (P < .05): right posterior sectorial duct joining left hepatic duct with a supraportal course (34% vs 12%), right posterior sectorial duct joining right anterior sectorial duct with an infraportal course (13% vs 4%), right posterior sectorial duct following an infraportal course (23% vs 8%), and left lateral segmental ducts caudal to the umbilical portion of the portal vein (14% vs 3%). The right hepatic duct, which receives all biliary ducts from the right lobe, was significantly less frequently developed in the variation group (46% vs 79%, P < .05). In addition, retroportal bile ducts were seen in four patients with the portal vein variation (P < .05). CONCLUSION: Bile duct configurations in patients with portal vein variation were significantly different from those in control subjects. PMID- 16373766 TI - Endorectal MR imaging before salvage prostatectomy: tumor localization and staging. AB - PURPOSE: To evaluate retrospectively the accuracy of endorectal magnetic resonance (MR) imaging for the depiction of tumor, extracapsular extension (ECE), and seminal vesicle invasion (SVI) before salvage prostatectomy in patients with locally recurrent prostate cancer after radiation therapy, by using pathologic analysis as the reference standard. MATERIALS AND METHODS: The Institutional Review Board granted exempt status for this HIPAA-compliant study, with a waiver of informed consent. Forty-five consecutive patients (age range, 43-76 years) were identified who underwent salvage radical prostatectomy for prostate cancer at Memorial Sloan-Kettering Cancer Center between December 1, 1998, and October 31, 2004, and who underwent endorectal MR imaging prior to surgery. Tumor localization and determination of local stage with MR imaging were performed independently by two radiologists. Interpretations were compared to pathologic findings from surgical specimens. Interrater variability was estimated with the kappa statistic. Areas under the receiver operating characteristic curve (AUCs) were used to assess the accuracy of endorectal MR imaging in tumor detection and determination of ECE and SVI. RESULTS: Findings of histologic examination showed that tumor was present in all patients. For tumor detection, the AUC value for reader 1 was 0.75 (95% confidence interval [CI]: 0.67, 0.84), whereas the AUC value for reader 2 was 0.61 (95% CI: 0.52, 0.71). The AUC values for prediction of ECE were 0.87 (95% CI: 0.80, 0.94) for reader 1 and 0.76 (95% CI: 0.67, 0.85) for reader 2. The AUC values for prediction of SVI were 0.76 (95% CI: 0.62, 0.90) for reader 1 and 0.70 (95% CI: 0.56, 0.85) for reader 2. For all variables, the kappa statistics used to assess interrater agreement between readers were fair (0.45, 0.52, and 0.47 for tumor location, ECE, and SVI, respectively). CONCLUSION: Endorectal MR imaging following radiation therapy can help identify tumor sites and depict ECE and SVI with reasonable accuracy in patients with recurrent prostate cancer. PMID- 16373767 TI - Prostate cancer: endorectal MR imaging and MR spectroscopic imaging--distinction of true-positive results from chance-detected lesions. AB - PURPOSE: To retrospectively investigate size criteria for the identification of chance-detected lesions at endorectal magnetic resonance (MR) imaging and MR spectroscopic imaging of prostate cancer. MATERIALS AND METHODS: Approval of the committee on human research and written informed consent were obtained. This study was HIPAA compliant. Endorectal MR imaging and MR spectroscopic imaging were performed with a 1.5-T MR imager in 48 men with a mean age of 59 years (age range, 47-75 years) prior to radical prostatectomy. Two independent readers recorded the size and location of all suspected peripheral zone tumor nodules on MR images alone and on images obtained with combined MR imaging and MR spectroscopic imaging. Nodules detected at MR imaging were classified as matched lesions if tumor was present in the same location at step-section histopathologic review. For all matched lesions, kappa values were calculated to examine agreement between measured and actual tumor size. Lesions that were overmeasured at MR imaging with a kappa value of less than 0.2 were considered chance-detected lesions. RESULTS: At MR imaging, two of 27 and four of 35 matched lesions for readers 1 and 2, respectively, were chance-detected lesions. The corresponding numbers of lesions at combined MR imaging and MR spectroscopic imaging were one of 21 and one of 31, respectively. In all but two cases, the measured diameter of chance-detected lesions was more than twice that of the diameter at histopathologic analysis. By using this diameter threshold to distinguish true positive results, the mean diameter of detected tumors at histopathologic analysis was 15 mm compared with 4 mm for both undetected and chance-detected tumors (P < .05). CONCLUSION: To ensure uniformity in the comparison of scientific studies, peripheral zone tumors detected at MR imaging and MR spectroscopic imaging of the prostate that are in the same location as tumors detected at histopathologic review should be considered chance-detected lesions if the MR transverse diameter is more than twice the histopathologic transverse diameter. PMID- 16373768 TI - MR evaluation of the glomerular homing of magnetically labeled mesenchymal stem cells in a rat model of nephropathy. AB - PURPOSE: To assess renal glomerular homing of intravenously injected superparamagnetic iron oxide (SPIO)-labeled mesenchymal stem cells (MSCs) at in vivo and ex vivo magnetic resonance (MR) imaging in an experimental rat model of mesangiolysis. MATERIALS AND METHODS: Animal procedures were performed in accordance with protocols approved by Institutional Animal Care and Use Committee. Fourteen rats were divided into two groups: one pathologic (n = 10), with persistent mesangiolysis following simultaneous injection of OX-7 monoclonal antibody and puromycin aminonucleoside in which 10(7) SPIO- and DiI-labeled MSCs were injected, and one control (n = 4). In vivo and ex vivo MR imaging examinations were performed with 4.7- and 9.4-T spectrometers, respectively, and T2*-weighted sequences. In vivo signal intensity variations were measured in the liver and kidney before and 6 days after MSC injection. Intrarenal signal intensity variations were correlated with histopathologic data by means of colocalization of DiI fluorescence, alpha-actin, and Prussian blue stain-positive cells. Histologic differences between the glomerular homing of MSCs in different kidney portions were correlated to the areas of MR signal intensity decrease with nonparametric statistical tests. RESULTS: On in vivo images, signal intensity measurements of pathologic kidneys following MSC injection did not show any signal intensity decrease (P = .7), whereas a 34% +/- 14 (mean +/- standard deviation) signal intensity decrease was observed in the liver (P < .01), where a substantial number of labeled cells were trapped. On ex vivo images, pathologic kidneys showed focal cortical (glomerular) areas of signal intensity loss, which was absent in controls. The areas of low signal intensity correlated well with alpha-actin and Prussian blue stain- and DiI-positive areas (P < .01), which indicates that MSCs specifically home to injured tissue. No MSCs were detected in the kidneys of control animals. CONCLUSION: Intravenously injected MSCs specifically home to focal areas of glomerular damage and can be detected at ex vivo MR imaging. PMID- 16373769 TI - Osteoporotic compression fractures: outcomes after single- versus multiple-level percutaneous vertebroplasty. AB - PURPOSE: To compare single- and multiple-level percutaneous vertebroplasty (PV) in terms of pain relief, activity level, and analgesic use in patients with osteoporotic vertebral compression fractures (VCFs). MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained, and the study was HIPAA compliant. One hundred seventy-three patients (mean age at treatment, 73.8 years +/- 11.9 [standard deviation]) with 422 symptomatic osteoporotic VCFs underwent 204 treatment sessions for over 4 years. Pain immediately before and after PV was measured by using a visual analogue scale (VAS). Pain degree, activity level, and analgesic use were assessed at 2 weeks and 1, 3, 6, 12, and 24 months after PV by using telephone interview questionnaires. Data were analyzed by using a combination of paired t tests, analysis of variance, contingency tables, and chi2 tests. RESULTS: Findings of 172 PV treatment sessions for 149 patients (mean age at treatment, 73.4 years +/- 12), 110 (74%) of whom were women, were assessed; 32 treatment cases were lost to follow-up or lost owing to death. A single fracture level was treated at 65 sessions; two fracture levels, at 52 sessions; and three or more fracture levels, at 55 sessions. The mean VAS pain score decreased significantly (P < .001), from 76 +/- 21 before to 19 +/- 27 immediately after PV. Of the outcomes reported at 24 months, 82% (64 of 78 treatment sessions) were marked to complete resolution of the initial pain, 51% were complete cessation of analgesic use, and 51% were increased activity levels. These results did not differ greatly over time or when stratified into groups according to the number of fracture levels treated. CONCLUSION: PV performed at a single fracture level and that performed at multiple fracture levels were equally effective in facilitating long-term pain relief, increased activity level, and decreased analgesic use in patients with osteoporotic VCFs. PMID- 16373770 TI - Characteristics of displaceable and nondisplaceable meniscal tears at kinematic MR imaging of the knee. AB - PURPOSE: To prospectively determine if kinematic magnetic resonance (MR) imaging of the knee may demonstrate displacement of menisci with tears and, if so, to characterize displaceable and nondisplaceable meniscal tears. MATERIALS AND METHODS: The study was approved by the hospital's review board, and informed consent was obtained. Forty-two patients (30 men, 12 women; mean age, 36.9 years) with 43 arthroscopically documented meniscal tears visible at 1.5-T MR imaging underwent kinematic MR imaging with an open-configuration 0.5-T MR imager with their knees in supine neutral, supine with 90 degrees flexion and external or internal rotation, and upright weight-bearing positions. Analysis of meniscal movement was performed in different knee positions in the coronal MR imaging plane. Meniscal displacement--that is, meniscal movement of 3 mm or more (in the medial direction for the medial meniscus, in the lateral direction for the lateral meniscus)--was compared with the patient's pain level as assessed with a visual analog scale by using analysis of variance. RESULTS: Between the different knee positions, meniscal displacement of 3 mm or more (displaceable meniscal tears) was noted in 18 (42%) of 43 menisci with tears. Simultaneous occurrence of grade II or III ipsilateral collateral ligament lesions was present in all 18 displaceable meniscal tears, whereas a normal-appearing collateral ligament or collateral ligament lesion (grade I) was present in 22 of 25 nondisplaceable tears (P < .05). Displaced menisci most commonly had complex, radial, or longitudinal tear configurations (16 of 18, 89%). Patients with displaceable meniscal tears had significantly more pain than did patients with nondisplaceable meniscal tears (P < .001), independent of the concomitant knee abnormalities. CONCLUSION: Displaceable meniscal tears usually have longitudinal, radial, or complex configurations; such tears are associated with substantial ipsilateral collateral ligament lesions and pain. PMID- 16373771 TI - Acute ischemic stroke: accuracy of diffusion-weighted MR imaging--effects of b value and cerebrospinal fluid suppression. AB - PURPOSE: To prospectively determine which diffusion-weighted magnetic resonance (MR) imaging technique (ie, conventional diffusion-weighted MR imaging [b = 1000 or 1500 sec/mm2] or fluid-inversion prepared diffusion [FLIPD] MR imaging [b = 1500 sec/mm2]) is most accurate in depicting acute ischemic stroke at 3 T. MATERIALS AND METHODS: The Health Research Ethics Board approved this study; written informed consent was provided by all participants or their surrogate. Diffusion-weighted MR imaging was performed in 75 consecutive patients (43 men, 32 women; mean age, 64.0 years) with acute ischemic stroke. Two experienced neuroradiologists determined the presence of hyperacute stroke lesions at diffusion-weighted MR imaging by locating areas of hyperintensity that corresponded to regions with a decreased diffusion coefficient. These findings were used as the reference standard. Four raters who were blinded to patient history assessed all images and apparent diffusion coefficient maps for the presence of changes that were consistent with acute ischemic stroke. Accuracy, sensitivity, specificity, negative predictive value, positive predictive value, and inter- and intrarater reliability scores were calculated for each technique. RESULTS: Specificity, positive predictive value, and accuracy were not significantly different among the techniques. FLIPD MR images obtained with a b value of 1500 sec/mm2 had decreased sensitivity for acute ischemic stroke (mean, 61.8%; 95% confidence interval [CI]: 55.4%, 67.9%) compared with conventional diffusion-weighted MR images obtained with a b value of either 1000 sec/mm2 (mean, 82.5%; 95% CI: 77.1%, 87.0%) or 1500 sec/mm2 (mean, 84.5%; 95% CI: 79.3%, 88.9%). FLIPD MR images also had decreased negative predictive value (mean, 96.5%; 95% CI: 95.7%, 97.2%) compared with conventional diffusion-weighted MR images obtained with a b value of either 1000 sec/mm2 (mean, 98.4%; 95% CI: 97.8%, 98.8%) or 1500 sec/mm2 (mean, 98.6%; 95% CI: 98.1%, 99.0%). Intra- and interrater reliability scores were generally excellent for all three techniques. CONCLUSION: FLIPD MR images obtained with a b value of 1500 sec/mm2 are less suitable for the detection of acute ischemic stroke owing to a decreased sensitivity and negative predictive value. The performance of the two conventional diffusion-weighted MR imaging techniques (b = 1000 and 1500 sec/mm2) was equivalent. PMID- 16373772 TI - Coexistence of microhemorrhages and acute spontaneous brain hemorrhage: correlation with signs of microangiopathy and clinical data. AB - PURPOSE: To evaluate prospectively with magnetic resonance (MR) imaging the coexistence of microhemorrhages (MHs) in white patients with acute spontaneous intraparenchymal hemorrhage (IPH) and acute ischemic stroke and to study the association with imaging findings of microangiopathy and various clinical data. MATERIALS AND METHODS: Before examinations, informed consents were signed by either the patient or a relative. The study was carried out with the approval of the local ethics committee. MR imaging was performed in 90 patients with acute stroke: 45 with acute spontaneous IPHs (24 men and 21 women; median age, 65 and 68 years, respectively) and 45 age-matched control subjects without intracranial hemorrhages (26 men and 19 women; median age for both, 67 years), as determined at computed tomography. MR imaging included transverse T1- and T2-weighted spin echo, transverse fluid-attenuated inversion recovery, transverse and coronal T2* weighted gradient-echo, and, in 50 patients, diffusion-weighted sequences. Presence of MHs and signs of microangiopathy, such as T2 hyperintensities or lacunae, were recorded in the white and deep gray matter. The relationships between MH and IPH and between MH and T2 hyperintensities were analyzed by means of regression analysis. Different clinical features, such as arterial hypertension or diabetes, were registered and correlated with the image findings by means of regression analysis. RESULTS: MHs were found in 64% of patients with IPH (29 of 45) and 18% of control subjects (eight of 45). A statistically significant relationship between MH and IPH was determined (P < .001). Among the 29 patients with IPH and MH, 24 (83%) had T2 hyperintensities and 13 (45%) had lacunae; among the 16 patients without MH, seven (44%) had T2 hyperintensities and three (19%) had lacunae. A relationship between MH and occurrence and extent of T2 hyperintensities was also identified (P < .001). There was no clear relationship with the clinical data studied. CONCLUSION: The results support a correlation between the presence of imaging signs of cerebral microangiopathy, clinically silent MHs, and acute IPHs. PMID- 16373773 TI - Depressive psychosis: clinical usefulness of MR spectroscopy data in predicting prognosis. AB - PURPOSE: To prospectively assess the usefulness of magnetic resonance (MR) spectroscopy data acquired before the initiation of medical therapy in predicting prognosis in patients with depressive psychosis. MATERIALS AND METHODS: All subjects gave written informed consent to an institutional committee for clinical research-approved study protocol. The clinical course after medication in 52 patients with depressive psychosis (age range, 52-78 years; 21 men, 31 women) was investigated. In all patients, MR spectroscopy was performed with a 1.5-T MR imaging unit before the initiation of medical therapy. Cerebrovascular lesions (CVLs), which appear as high-signal-intensity areas on T2-weighted MR images, were evaluated by using the Fazekas rating scale. Patients were classified into two groups on the basis of the ratio of N-acetylaspartate (NAA) to creatine and phosphocreatine (Cr): Patients in group A had an NAA/Cr ratio greater than 1.91, and patients in group B had an NAA/Cr ratio of 1.91 or less. To assess the response of the patients to medication, standard psychiatric tests--the Verbal Associative Fluency Test (VAFT), the Digit Symbol Test (DST), the Mini-Mental State Examination (MMSE), and the Hamilton Depression Rating Scale (HAM-D)--were administered before and after medical therapy was initiated. Mean test scores before and after medication were compared with paired t testing. P < .05 was considered to indicate a significant difference. RESULTS: There were 25 patients in group A and 27 in group B. In group A, the mean VAFT and DST scores increased and the mean HAM-D score decreased after medication. There was no significant difference in mean MMSE scores before and after medication (P = .945 for group A and P = .934 for group B). In group B, there were no significant differences in any of the psychiatric test scores before and after medication. The high-signal intensity area score in group B was significantly higher than that in group A (P = .004). CONCLUSION: MR spectroscopy data obtained before the initiation of medical therapy were useful in predicting prognosis in patients with depressive psychosis; this suggests that the combined burden of all CVLs may affect the response to antidepressant medication. PMID- 16373774 TI - Optimization of multiplanar reformations from isotropic data sets acquired with 16-detector row helical CT scanner. AB - Institutional review board approval and waiver of consent were obtained for the patient component of this retrospective HIPAA-compliant study. By using an anthropomorphic phantom and metal oxide semiconductor field effect transistor detectors, radiation dose was determined for one eight-detector row and two 16 detector row computed tomographic (CT) protocols. A custom phantom was scanned by using the three protocols to identify isotropy. Contrast-to-noise ratios (CNRs) were determined for the same protocols by using a third phantom. Seven patients had undergone isotropic 16-detector row CT of the abdomen and pelvis. Anonymized coronal reformations at various thicknesses were ranked qualitatively by three radiologists. Effective dose equivalents were similar for the eight- and 16 detector row protocols. When transverse and coronal reformations of data acquired in the custom phantom were compared, coronal reformations obtained with the 16 detector row and 0.625-mm section thickness protocol were found to be nearly identical to the transverse image for all sets of line pairs. CNRs were consistently highest on 5-mm-thick coronal reformations (CNR range, 1.2-3.3). For qualitative assessment, 2- and 3-mm-thick coronal reformations were consistently preferred. PMID- 16373775 TI - Carotid artery stent implantation: evaluation with multi-detector row CT angiography and virtual angioscopy--initial experience. AB - Approval for this HIPAA-compliant study was obtained from the institutional review board; informed consent was not required for retrospective review of patient studies that had been performed for clinical evaluation. The purpose of this study was to retrospectively compare the accuracy of intrastent luminal diameter, as measured on transverse computed tomographic (CT) angiograms and virtual angioscopic views, with the manufacturer's specifications for phantom diameter and with digital subtraction angiographic (DSA) measurements of stent diameter obtained in patients. Intrastent diameter was measured by using standard and stent-optimized reconstruction kernels with three window settings. Endoluminal virtual angioscopic views of the stent-containing vessels were also generated. Measurements at CT angiography were compared with known specifications for the phantom and with DSA measurements in patients. Erroneous measurements of intrastent diameter occurred when a standard kernel and nonoptimized window settings were used. A set of parameters that minimized error relative to measurements obtained at DSA was also identified. Virtual angioscopy helped demonstrate morphologic aspects of stenosis that were otherwise difficult to appreciate. PMID- 16373776 TI - Radiofrequency ablation for small hepatocellular carcinoma: prospective comparison of internally cooled electrode and expandable electrode. AB - PURPOSE: To prospectively compare the effectiveness of radiofrequency (RF) ablation performed by using an internally cooled electrode and an expandable electrode for the treatment of small (< or = 3.0 cm) hepatocellular carcinomas (HCCs). MATERIALS AND METHODS: The human subjects research review board at the study institution approved the protocol, and each patient provided informed consent. Seventy-four patients (58 men and 16 women; age range, 41-83 years) with 83 HCC nodules 3 cm or smaller were randomly divided into an internally cooled electrode group (38 patients with 41 nodules) and an expandable electrode group (36 patients with 42 nodules). RF ablation was performed by one individual. Primary technique effectiveness and rates of major complications were evaluated between the two groups with the Fisher exact test. Rates of local tumor progression, overall survival, local progression-free survival, and event-free survival were evaluated by using the Kaplan-Meier method. RESULTS: The primary technique effectiveness was 95% in the internally cooled electrode group and 93% in the expandable electrode group (P = .51); rates of major complications were 0% and 2.1% per session (P = .50) and 0% and 2.8% per patient (P = .49), respectively. Rates at 1, 2, and 3 years in the internally cooled electrode group versus the expandable electrode group were as follows: local tumor progression, 12% versus 17%, 20% versus 22%, and 20% versus 22% (P = .72, log-rank test); overall survival, 100% versus 94%, 94% versus 92%, and 94% versus 77% (P = .29, log-rank test); local progression-free survival, 87% versus 78%, 73% versus 66%, and 73% versus 46% (P = .27, log-rank test); and event-free survival, 47% versus 44%, 34% versus 22%, and 34% versus 22% (P = .40, log-rank test). CONCLUSION: On the basis of the study findings, RF ablation with an internally cooled electrode needle and an expandable electrode needle has equivalent effectiveness for the treatment of small HCCs. PMID- 16373777 TI - Percutaneous vertebroplasty for spinal metastases: complications. AB - PURPOSE: To retrospectively evaluate complications of percutaneous vertebroplasty (PV) performed with polymethylmethacrylate cement to treat pain in patients with metastases to the spine. MATERIALS AND METHODS: This study had institutional review board approval; patient informed consent for the review of records and images was not required. In 2 years, 117 patients (38 men [32.5%] and 79 women [67.5%]; mean age, 58.2 years) underwent 159 fluoroscopy-guided PV procedures to treat 304 vertebrae. Spinal metastases included osteolytic, osteoblastic, and mixed lesions. Complications were characterized as local or systemic. Evaluated data included immediate imaging findings (on radiographs and computed tomographic scans) and clinical findings at 30-day follow-up. Chi2 or Fisher exact testing was performed for univariate analysis of variables. RESULTS: The primary cancers were breast cancers (45.3%), lung cancers (14.5%), myeloma (7.7%), or other cancers (32.5%). Among the 423 cement leakages identified, 332 (78.5%) were vascular and 91 (21.5%) were nonvascular. Vascular leaks were classified as venous epidural leaks, paravertebral and foraminal plexus leaks, and leaks to the vena cava, while nonvascular leaks included puncture trajectory leaks, paravertebral soft tissue leaks, and diskal leaks. Patients with nonvascular leaks were asymptomatic. Eight (6.8%) patients experienced complications, and seven of these complications were symptomatic. Among these eight patients, six (5.1%) had local complications (puncture site hematoma in two patients and radicular pain [successfully treated with nonsteroidal anti-inflammatory drugs or corticosteroids] in four patients), and two (1.7%) had systemic complications (pulmonary embolism resulting from cement migration through the vena cava). One of the latter patients died. Univariate analyses revealed a significant association between cement migration through the vena cava and pulmonary embolism (P = .001) but not between foraminal venous leakage and radicular pain (P = .123). CONCLUSION: Despite numerous technical incidents (leaks), PV-induced complications were rare, leading to the hypothesis that systemic complications are a consequence of intravascular leakage while local complications are a consequence of cement-related irritation, compression and/or ischemia, and/or needle-induced trauma. PMID- 16373779 TI - Case 90: disseminated tuberculosis. PMID- 16373780 TI - The bridging vascular sign. PMID- 16373781 TI - Enhancement characteristics of pheochromocytomas. PMID- 16373782 TI - Prediction of pulmonary function in COPD on the basis of CT measurements of bronchial wall thickness. PMID- 16373783 TI - Regarding trends in recall, biopsy, and positive biopsy rates for screening mammography. PMID- 16373784 TI - Attenuation of acute and chronic clots. PMID- 16373786 TI - Vertebral artery dissection and cerebellar infarction following chiropractic manipulation. AB - Vertebral artery dissection (VAD) associated with chiropractic cervical manipulation is a rare but potentially disabling condition. In this report, we present a young patient manifesting with repeated vertigo. Owing to the initial misdiagnosis, the patient later developed cerebellar stroke with inability to stand or walk. Vertigo and disequilibrium are the usual presenting symptoms of this condition, which can result from inner ear or vestibular nerve dysfunction, vertebrobasilar insufficiency, and even lethal cerebellar infarction or haemorrhage; these last two, although rarely seen in young adults, can be caused by traumatic or spontaneous arterial injury, including injury secondary to chiropractic cervical manipulation. A number of cases of VAD associated with chiropractic cervical manipulation have been reported, but rarely in the emergency medicine literature. We present a case of this rare occurrence, and discuss the diagnostic pitfalls. PMID- 16373787 TI - How ECG can cause confusion in pulmonary embolism and how echocardiogram can help. AB - A case of pulmonary embolism in which the diagnosis was aided by transthorasic echocardiography is described. Echocardiography may be helpful in emergency presentations, as ECG changes can be neither sensitive nor specific for the diagnosis of acute massive pulmonary embolism. PMID- 16373788 TI - Aluminium phosphide poisoning. AB - We describe a lethal poisoning in a healthy woman caused by deliberate ingestion of aluminium phosphide (AlP), a pesticide used to kill rodents and insects. Toxicity of AlP and review of cases reported to the National Poisons Information Service (London) 1997-2003 are discussed. PMID- 16373789 TI - Factitious buccal lesion secondary to bruxism in a child with cerebral palsy. AB - Children with cerebral palsy are at greater risk of a whole range of oral conditions than their peers. These include bruxism (tooth grinding), oral skill dysfunction, gross malocclusion due to effects of the abnormal orofacial muscle tone on tooth eruption, drooling of saliva, and poor oral hygiene. A challenging case of a painful buccal lesion in a 2 year old girl with cerebral palsy (CP) that did not respond to antifungal, antiviral or antibiotic treatment is presented as a factitious lesion. The recognition and significance of self injurious behaviour and factitious lesions in children are discussed. PMID- 16373790 TI - Is the patient brain-dead? PMID- 16373791 TI - Noninvasive ventilation in acute neuromuscular respiratory failure due to myasthenic crisis: case report and review of literature. PMID- 16373792 TI - Hypopharyngeal perforation following minor trauma: a case report and literature review. AB - Hypopharyngeal perforation is mainly reported in association with high velocity road traffic accidents, or with low velocity direct blows to the neck. We report a case of hypopharyngeal perforation following a low velocity motorcycle accident where neither of these mechanisms of injury was apparent. A 52 year old man was referred from the emergency department (ED) with a sore throat and dysphagia, following a low speed side impact accident. A gastrograffin swallow demonstrated a posterior pharyngeal wall tear. After 11 days of conservative hospital treatment, he recovered and was discharged. The presumed mechanism of injury in this case was cervical spine hyperextension without cervical compression. PMID- 16373793 TI - Two cases of retroperitoneal haematoma caused by interaction between antibiotics and warfarin. AB - Several commonly prescribed antibiotics are known to interact with warfarin, increasing its anticoagulant effect by different mechanisms. Retroperitoneal bleeding with consequent haematoma is recognised as a complication of over anticoagulation. Consequences, which are potentially fatal, include hypovolaemic shock and compression of retroperitoneal structures such as the ureter and inferior vena cava. PMID- 16373794 TI - The four hour target; problems ahead. PMID- 16373795 TI - Potential cervical spine injury and difficult airway management for emergency intubation of trauma adults in the emergency department--a systematic review. AB - BACKGROUND: Emergency airway management for trauma adults is practised by physicians from a range of training backgrounds and with differing levels of experience. The indications for intubation and technique employed are factors that vary within EDs and between hospitals. OBJECTIVES: To provide practical evidence based guidance for airway management in trauma resuscitation: first for the trauma adult with potential cervical spine injury and second the management when a difficult airway is encountered at intubation. SEARCH STRATEGY AND METHODOLOGY: Full literature search for relevant articles in Medline (1966-2003), EMBASE (1980-2003), and the Cochrane Central Register of Controlled Trials. Relevant articles relating to adults and written in English language were appraised. English language abstracts of foreign articles were included. Studies were critically appraised on a standardised data collection sheet to assess validity and quality of evidence. The level of evidence was allocated using the methods of the Australian National Health and Medical Research Council. PMID- 16373796 TI - Tackling alcohol related violence in city centres: effect of emergency medicine and police intervention. AB - OBJECTIVES: To identify correlates of alcohol related assault injury in the city centre of a European capital city, with particular reference to emergency department (ED) and police interventions, and number and capacity of licensed premises. METHODS: Assaults resulting in ED treatment were studied using a longitudinal controlled intervention, a three stage design during a three year period of rapid expansion in the night-time economy, when ED initiated targeted police interventions were delivered. A controlled ED intervention targeted at high risk night-clubs was carried out. Main outcome measure was ED treatment after assault in licensed premises and the street. RESULTS: Targeted police intervention was associated with substantial reductions in assaults in licensed premises but unexpected increases in street assault were also observed (34% overall: 105% in the principal entertainment thoroughfare). Combined police/ED intervention was associated with a significantly greater reduction compared with police intervention alone (OR = 0.61, 95% CI 0.40 to 0.91). Street assault correlated significantly with numbers and capacity of premises. Risk of assault was 50% greater in and around licensed premises in the city centre compared with those in the suburbs, although dispersion of violence to more licensed premises was not observed. CONCLUSIONS: Marked decreases in licensed premises assaults resulting from targeted policing were enhanced by the intervention of ED and maxillofacial consultants. Capacity of licensed premises was a major predictor of assaults in the city centre street in which they are clustered. City centre assault injury prevention can be achieved through police/ED interventions targeted at high risk licensed premises, which should also target the streets around which these premises are clustered. PMID- 16373797 TI - A structured competency based training programme for junior trainees in emergency medicine: the "Dundee Model". AB - Recent changes in medical training prompted by Modernising Medical Careers and the New Deal requires a more structured, competency based training programme. This paper describes the development of such a programme in an emergency medicine department of a teaching hospital. It describes the process of design and the various aspects incorporated to develop a balanced system of training, appraisal, and assessment. PMID- 16373798 TI - Potential health promotion benefits of lipid testing for all patients presenting with chest pain to an emergency department. AB - OBJECTIVES: To establish the prevalence of previously undiagnosed dyslipidaemia in patients presenting to the emergency department (ED) with non-traumatic chest pain and, more particularly, the prevalence in the subgroup which was discharged home from the ED, the group that traditionally would not have received a lipid test. METHODS: Prospective, observational study of adult patients presenting to an ED with non-traumatic chest pain as the presenting complaint. RESULTS: A total of 185 eligible patients underwent lipid testing during their presentation: 96 in the ED and 89 in the wards. Overall 61% (n = 112) of patients had at least one abnormal lipid level. Of patients discharged from the ED, 62% had at least one abnormal lipid level. CONCLUSIONS: A moderate, but useful, increase in detection rates of dyslipidaemia is possible if lipid testing is offered to all patients presenting with chest pain, and not just to those who are admitted to wards for further investigation and management of suspected acute coronary syndromes. Testing of this group should be considered as a health promotion initiative in the ED, with appropriate follow up in the community. PMID- 16373799 TI - Motor vehicle trauma: analysis of injury profiles by road-user category. AB - BACKGROUND: Vehicle accidents in Greece are among the leading causes of death and the primary one in young people. The mechanism of injury influences the patterns of injury in victims of vehicle accidents. OBJECTIVE: Identification and analysis of injury profiles of motor-vehicle trauma patients in a Greek level I trauma centre, by road-user category. PATIENTS AND METHODS: The trauma registry data of Herakleion University Hospital of adult trauma patients admitted to the hospital after a vehicle accident between 1997 and 2000 were retrospectively examined. Patients were grouped based on the mechanism of injury into three road-user categories: car occupants, motorcyclists, and pedestrians. RESULTS: Of 730 consecutive patients, 444 were motorcyclists (60.8%), 209 were car occupants (28.7%), and 77 were pedestrians (10.5%). Young men constituted the majority of injured motorcyclists whereas older patients (p = 0.0001) and women (p = 0.0001) represented a substantial proportion of the injured pedestrians. With regard to the spectrum of injuries in the groups, craniocerebral injuries were significantly more frequent in motorcyclists and pedestrians (p = 0.0001); abdominal (p = 0.009) and spinal cord trauma (p = 0.007) in car occupants; and pelvic injuries (p = 0.0001) in pedestrians. Although the car occupants had the highest Injury Severity Score (ISS) (p = 0.04), the pedestrians had the poorest outcome with substantially higher mortality (p = 0.007) than the other two groups. CONCLUSIONS: The results reveal a clear association between different road-user categories and age and sex incidence patterns, as well as outcomes and injury profiles. Recognition of these features would be useful in designing effective prevention strategies and in comprehensive prehospital and inhospital treatment of motor-vehicle trauma patients. PMID- 16373800 TI - Streptococcus A in paediatric accident and emergency: are rapid streptococcal tests and clinical examination of any help? AB - BACKGROUND: Rapid streptococcal tests (RSTs) for streptococcal pharyngitis have made diagnosis at once simpler and more complicated. The American Academy of Pediatrics recommends that all RSTs be confirmed by a follow up throat culture unless local validation has proved the RST to be equally sensitive. AIMS: To evaluate (a) RST as a single diagnostic tool, compared with RST with or without throat culture; (b) clinical diagnosis and the relative contribution of different symptoms. METHODS: The study included 213 patients with clinical signs of pharyngitis. Throat swabs were analysed using Quickvue+ Strep A Test; negative RSTs were backed up by throat culture. Thirteen clinical features commonly associated with strep throat were analysed using backward stepwise logistic regression. RESULTS: Positive results (RST or throat culture) were obtained in 33 patients; RST correctly identified 21. Eleven samples were false negative on RST. At a strep throat prevalence of 15.9%, sensitivity of RST was 65.6% (95% CI 46.8% to 81.4%) and specificity 99.4% (96.7% to 99.9%). Sensitivity of clinical diagnosis alone was 57% (34% to 78%) and specificity 71% (61% to 80%). Clinically, only history of sore throat, rash, and pyrexia contributed to the diagnosis of strep throat (p<0.05). CONCLUSION: The high specificity of RST facilitates early diagnosis of strep throat. However, the low sensitivity of RST does not support its use as a single diagnostic tool. The sensitivity in the present study is markedly different from that reported by the manufacturer. Clinical examination is of limited value in the diagnosis of strep throat. It is important to audit the performance of new diagnostic tests, previously validated in different settings. PMID- 16373801 TI - Effectiveness of measures to reduce emergency department waiting times: a natural experiment. AB - OBJECTIVES: To determine what measures were introduced by emergency departments in response to the national monitoring week in March 2003, and which, if any, of these were most effective in reducing waiting times. METHODS: A postal survey of all emergency departments in England was undertaken to gather data on measures taken. Department waiting times before, during, and after monitoring week were determined from data held by the Department of Health and linked to the survey data for analysis. RESULTS: A total of 111/198 responses (56%) were received. Departments had taken a wide range of measures to improve waiting times. The commonest were additional senior doctor hours (39%), creation of a "four hour monitor" role (37%), improved access to emergency beds (36%), additional non clinical staff hours (33%), additional junior doctor hours (32%), additional nursing hours (29%), and triage by senior staff (28%). In 35 departments (32%) no changes were made at all to usual practice. The biggest influence on improved performance during monitoring week was the number of measures that a department took, rather than any specific measure, although there was weak evidence that additional junior medical and non-clinical staff time may have contributed more than other measures. CONCLUSIONS: Improved waiting time performance may depend, at least in the short term, more on the amount of effort expended than on introducing a single effective change. In addition, those measures most likely to be helpful are likely also to require additional resources. PMID- 16373802 TI - An evaluation of tympanic thermometry in a paediatric emergency department. AB - BACKGROUND AND OBJECTIVES: The consequences of missing fever in children can be grave. Body temperature is commonly recorded at the axilla but accuracy is a problem. This study aimed to evaluate the accuracy of a tympanic thermometer in the paediatric emergency setting. METHOD: In a total of 106 infants, the body temperature was measured in the daytime with an infrared tympanic thermometer, and at the axilla with an electronic thermometer and at the rectum (gold standard for measurement of body temperature). Fever was defined as a rectal temperature of 38.0 degrees C or greater, axillary temperature of 37.5 degrees C or greater, and tympanic temperature of 37.6 degrees C or greater. The temperature readings at the three sites were compared statistically. RESULTS: There was a greater correlation of the tympanic measurement with the rectal measurement than the axillary with the rectal in both febrile and afebrile infants. The mean difference between the tympanic and rectal measurements was lower than that between the axillary and rectal measurements in both groups of infants (tympanic 0.38 degrees C and 0.42 degrees C, and axillary 1.11 degrees C and 1.58 degrees C, respectively). Tympanic measurements had a sensitivity of 76% whereas axillary measurements had a sensitivity of only 24% with rectal temperatures of 38-38.9 degrees C. CONCLUSION: Tympanic thermometry is more accurate than measurement of temperature with an electronic axillary thermometer. It is also quick and safe, and thus it is recommended in the paediatric emergency setting. PMID- 16373803 TI - Validation of the Paediatric Triage Tape. AB - INTRODUCTION: The Paediatric Triage Tape (PTT) is an easy to use major incident primary triage tool, based upon a modification of the Triage Sieve. The purpose of this study was to prospectively validate the PTT for use in paediatric major incidents. METHODS: A database of children presenting the Trauma Unit of the Red Cross Children's Hospital, Cape Town, was developed over a nine month period. Each child was triaged using the PTT, and had an Injury Severity Score (ISS) calculated. Additionally, the New Injury Severity Score (NISS) was calculated, and the presence of interventions that may occur to the children ("Garner criteria") was documented. The sensitivity, specificity, overtriage, and undertriage rates were calculated. RESULTS: 3461 children were entered into the database. For identifying children with an ISS of over 15, the PTT had a sensitivity of 37.8%, specificity of 98.6%, overtriage rate of 38.8%, and an undertriage rate of 3.5%. Against the NISS and Garner criteria, the results were comparable. CONCLUSION: The PTT has poor sensitivity at identifying immediate priority children by these criteria. Specificity (the ability to identify non-T1 patients) is excellent, and the overtriage and undertriage rates are within the range deemed unavoidable by the American College of Surgeons. PMID- 16373804 TI - Biphasic cardioversion of acute atrial fibrillation in the emergency department. AB - INTRODUCTION: There is a trend towards accelerated management of acute atrial fibrillation (AF) in the emergency department (ED). We report our experience with biphasic cardioversion of acute AF. METHODS: This was a prospective, descriptive study at a tertiary hospital ED over a 6 month period. Acute AF was defined as symptoms that had been present for <48 hours. Patients who received biphasic cardioversion for acute AF in the ED were enrolled. Data collected included: patient demographics, past medical history, details of biphasic cardioversion, outcome, complications, disposition, and length of stay. RESULTS: There were 34 attempts at cardioversion in 33 patients. The mean (SD) age was 56 (16) years and 21 patients (64%) were men. Biphasic cardioversion was successful in 31 attempts (91%). In 24 attempts (71%), 100 J was selected as the initial energy level. This was successful in 21 attempts (88%). There were three minor complications related to sedation. The mean (SD) length of stay was 5.6 (2.8) hours in the ED and 15 (25) hours in the hospital. The three patients who failed to revert were older (mean age 64 years), had underlying cardiovascular disease, and spent longer in hospital (50 v 12 hours, p = 0.01). Telephone follow up was conducted with 32 patients (97%) at 3 months. Recurrence of AF occurred in 7 patients (22%). Most patients (31, 97%) were satisfied with the biphasic cardioversion. CONCLUSIONS: Biphasic cardioversion of acute AF is effective. The majority of patients can be managed as outpatients, and there is very high patient satisfaction with this approach. An initial shock energy level of 100 J is usually effective. PMID- 16373805 TI - Rapid sequence intubation in the emergency department: 5 year trends. AB - AIM: Airway management is a core aspect of emergency medicine. The technique of rapid sequence intubation (RSI) creates continuing debate between anaesthetists and emergency physicians in the UK, although similar complication rates for emergency department (ED) RSI have been shown for both specialties. This study examined prospectively collected data on every ED RSI performed in a university hospital in Glasgow over 5 years. METHODS: Data were prospectively recorded for every attempted RSI in the ED on a dedicated form (as used in previous studies) between January 1999 and December 2003. Immediate complications were specifically sought in the questionnaire, as was the immediate destination on leaving the ED. The chi2 test was used for categorical data. RESULTS: On average, 51 ED RSI were performed annually (range 42-60). Emergency physician RSI for trauma increased from 32% (7/22) in 1999 to 75% (21/28) in 2003 (chi2 = 9.32, df = 1, p = 0.002) and for non-trauma from 62% (18/29) in 1999 to 79% (23/29) in 2003 (chi2 = 2.08, df = 1, p = 0.15). Complication rates for emergency physician RSI decreased from 43% (3/7) to 14% (3/21) for trauma (chi2 = 2.55, df = 1, p = 0.11) and from 28% (5/18) to 4% (1/23) for non-trauma (chi2 = 4.44, df = 1, p = 0.035). This compares with mean complication rates for anaesthetists for trauma of 17% and for non-trauma of 22%. Incidence of hypotension decreased in all groups; however, oxygen desaturation is now the most common complication. The rate of ED RSI prior to computed tomography (CT) scans increased in both the trauma (79% v 42%; chi2 = 7.42, df = 1, p = 0.0065) and non-trauma (48% v 17%; chi2 = 5.85, df = 1, p = 0.016) groups. CONCLUSION: Emergency physician performed ED RSI is increasingly common but is not associated with overall higher numbers of RSIs being performed in the ED. Effective pre-oxygenation should be emphasised during training. PMID- 16373806 TI - Evaluation of propofol and remifentanil for intravenous sedation for reducing shoulder dislocations in the emergency department. AB - OBJECTIVES: To assess the combination of propofol and remifentanil for sedation to reduce shoulder dislocations in an ED. METHODS: Eleven patients with anterior glenohumeral dislocation were given propofol 0.5 mg/kg and remifentanil 0.5 microg/kg iv over 90 seconds and then further doses of 0.25 mg/kg and 0.25 microg/kg, respectively, if needed. Another practitioner attempted reduction using the Milch technique. RESULTS: Reduction was achieved in all patients within four minutes of giving sedation (range 0.3-4; mean 1.6). Seven required one attempt at shoulder reduction, three required two attempts, and one required three attempts. Mean time to recovery of alert status was three minutes (range 1 6). The mean pain score during the reduction was 1.7 out of 10 (range 0-5). Nine patients had full recall, one had partial recall, and one had no recall at all. Eight patients were "very satisfied" with the sedation and three were "satisfied". There were no respiratory or haemodynamic complications that required treatment. CONCLUSIONS: Propofol and remifentanil provide excellent sedation and analgesia for the reduction of anterior glenohumeral dislocation, enabling rapid recovery. PMID- 16373807 TI - Injury patterns in children with frequent emergency department visits. AB - OBJECTIVES: To compare injury patterns in children with many and few emergency department (ED) visits in order to reveal the causes for the frequent visits. METHODS: Three cohorts of Danish children (total 579 721 children) were followed for three years when their ages were 0-2, 6-8, and 12-14 years. Information on all ED visits was obtained from the Danish National Patient Registry. Injury type, place of accident, injury mechanism, admission, and distance to ED were compared between children with frequent ED visits (five or more during the three years) and children with only one visit. RESULTS: Children with frequent visits had a different injury pattern with 0-46% more superficial injuries and 25-82% more dislocations, sprains, and strains. There was 20-30% fewer fractures and 12% fewer falls from a higher level. 15-51% fewer were admitted. CONCLUSIONS: Children with many ED visits had less severe injuries and more dislocations, sprains, and strains. PMID- 16373808 TI - Best evidence topic report. Use of intravenous cyclizine in cardiac chest pain. AB - A short cut review was carried out to establish whether cyclizine adversely affected haemodynamic parameters in patients with cardiac disease. A total of 70 papers were found of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of this best paper are tabulated. The clinical bottom line is that cyclizine should be avoided in patients with acute coronary events. PMID- 16373810 TI - Best evidence topic report. Rapid sequence induction in the emergency department by emergency medicine personnel. AB - A short cut review was carried out to establish whether there are significant differences in the performance of emergency physicians and anaesthetists when carrying out rapid sequence intubation (RSI) in the emergency department. A total of 407 papers were found of which 12 presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. The clinical bottom line is that there is little or no difference in the rates of success and complications between emergency department clinicians and anaesthetists performing RSI. PMID- 16373811 TI - Best evidence topic report. Absorbable sutures in paediatric lacerations. AB - A short cut review was carried out to establish whether absorbable sutures offered any benefits over non-absorbable sutures in the treatment of childhood facial lacerations. A total of 31 papers were found, of which one presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of this best paper are tabulated. We conclude that absorbable sutures appear to be as good as, and show a trend towards benefit in the treatment of paediatric lacerations. PMID- 16373812 TI - Best evidence topic report. Smectite for acute diarrhoea in children. AB - A short cut review was carried out to establish whether smectite was a useful therapy in acute diarrhoea. A total of 21 papers were found of which five presented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. The clinical bottom line is that oral smectite appears to be effective at shortening the duration of the diarrhoea in children with acute diarrhoea rehydrated with oral rehydration solution. PMID- 16373813 TI - Use of a forensic technique to identify blood contamination of emergency department and ambulance trauma equipment. AB - Using a Kastle-Meyer (KM) technique, the following equipment from the emergency departments of six UK hospitals (four trusts) and three regional ambulance services was tested for blood contamination: extrication ("spinal") boards, cervical collars, straps, box splints, head blocks, and headboards. Only equipment ready for patient use was tested. Over half of trauma equipment (57%) tested positive for blood, including 15% of equipment that was visibly stained with blood. There have been no recorded cases of infection from contaminated trauma equipment but our study has identified the potential risk. Disposable covers for boards, disposable straps, and disposable radiolucent head blocks which are currently available provide a solution but have resource implications. PMID- 16373814 TI - Establishing a rural Emergency Medical Retrieval Service. AB - In 2004 the Argyll and Clyde health board established the Emergency Medical Retrieval Service to support its rural community hospitals. This article describes both why the service was established and its aims. This service covers a geographically extensive area, with approximately 85,000 people living in remote locations. Rural general practitioners in six community hospitals provide initial patient assessment and resuscitation. Providing emergency care and safe transfer of seriously ill and injured patients presenting to these community hospitals is a significant challenge. All parties involved felt that there was a need to provide a service to transport critically ill and injured patients from these remote locations to definitive care. The idea of the team is to bring the resuscitation room to the patient in the rural setting. With this aim and in order to implement the Intensive Care Society guidelines for the transport of critically ill patients, it was decided that consultants in Emergency Medicine and Anaesthetics with an interest in critical care would staff the service medically. This service is unique within the UK and the authors aim to report our findings from ongoing research and audit in future papers. PMID- 16373815 TI - Does initiation of an ambulance pre-alert call reduce the door to needle time in acute myocardial infarct? AB - OBJECTIVES: To assess the effect an ambulance pre-alert call for patients with suspected acute myocardial infarction (AMI) would have on door to needle (DTN) times. METHODS: We carried out back to back audits of DTN times following the initiation of the pre-alert calls. PARTICIPANTS: All patients thrombolysed within the emergency department between July 2003 and April 2004 (inclusive). STATISTICAL ANALYSIS: Mean DTN times and time to ECG pre-change and post-change were compared using the Two sample t test. The Fisher's exact test was used to compare pre-change and post-change proportions of patients seen within guideline times. RESULTS: In total, 73 patients were thrombolysed with 40 of these arriving by ambulance. Eighteen of these 40 were pre-change and 22 were post-change. Four patients were excluded. Fifty per cent of the pre-change group had a DTN time of <30 minutes compared with 91% of the post-change group (p = 0.005, Fisher's exact test). The phase one mean DTN time was found to be significantly greater than that for phase two (Two sample t test, p = 0.016; 95% CI 1.6 to 14.6). CONCLUSIONS: There was a significant reduction in DTN times after the introduction of the pre-alert call. PMID- 16373816 TI - Case of the month: rivastigmine (Exelon) toxicity with evidence of respiratory depression. AB - Rivastigmine, which has been approved by the US Food and Drugs Administration for the treatment of Alzheimer's disease, is a non-competitive reversible inhibitor of acetylcholinesterase. We present a case of rivastigmine toxicity at a dose of 90 mg, with evidence of respiratory depression. To our knowledge, this case report provides evidence of the highest rivastigmine ingestion recorded (90 mg) that caused respiratory depression but requiring only supportive intervention without the need for ralidoxime. Emergency physicians should strongly consider cholinesterase inhibitor (rivastigmine, galantamine, and tacrine) ingestion in patients who present with short and temporary organophosphate-like toxidromes. PMID- 16373818 TI - Images in emergency medicine: Acute severe coronary spasm associated with 5 fluorouracil chemotherapy. PMID- 16373819 TI - Phenotypic characterization of pulmonary arteries in normal and diseased lung. AB - Vascular endothelium is a continuous cell layer lining the cardiovascular system and serves as an interface between blood and the vascular wall tissue. Although the basic morphology of endothelial cells is similar in blood vessels of different organs and tissues, there is a great heterogeneity in endothelial cell types based on structural, metabolic, and developmental differences within each organ, particularly in the pulmonary vasculature. Current data about the usage of different markers for the immunohistochemical detection of endothelial cells in lung tissue are summarized, and functional aspects of caveolin expression after lung injury and in pulmonary hypertension are discussed. PMID- 16373820 TI - Serotonin transporter and receptors in various forms of human pulmonary hypertension. PMID- 16373821 TI - Alveolocapillary cross-talk: Giles F. Filley lecture. PMID- 16373822 TI - Suppression of type II bone morphogenic protein receptor in vascular smooth muscle induces pulmonary arterial hypertension in transgenic mice. PMID- 16373823 TI - Superoxide and chronic hypoxia-induced pulmonary hypertension in newborn piglets. PMID- 16373824 TI - Redox activation of intracellular calcium release channels (ryanodine receptors) in the sustained phase of hypoxia-induced pulmonary vasoconstriction. AB - Hypoxia-induced pulmonary vasoconstriction (HPV) is an important adaptive process that remains incompletely understood. In preconstricted rat pulmonary arteries (inner diameter, 250 to 400 microm), hypoxia (pO2 approximately 10 mm Hg) induces an initial transient phase and a more slowly developing sustained phase of vasoconstriction. Since the release of calcium ions (Ca2+) from intracellular stores by redox-sensitive intracellular Ca2+ release channels known as ryanodine receptors (RyRs) in pulmonary arterial smooth-muscle cells (PASMCs) may play a role in HPV, and considerable evidence now supports that levels of reactive oxygen species (ROS) are paradoxically increased in PASMC under hypoxia, we investigated whether redox activation of RyRs by ROS may transduce HPV. By reverse transcriptase-polymerase chain reaction, we found that all three RyR isoforms are expressed in rat pulmonary arteries and in PASMCs. The sustained phase, but not the transient phase, of HPV can be prevented by pretreating pulmonary arteries with RyR inhibitors ryanodine (200 micromol/L) or dantrolene (50 micromol/L). The addition of dantrolene, ryanodine or the thiol-reducing agent dithiothreitol (1 mmol/L) during the sustained phase of HPV reversed the hypoxic vasoconstriction. In contrast, the superoxide scavenger nitroblue tetrazolium (500 nmol/L) prevented further hypoxic pulmonary vasoconstriction during the sustained phase of HPV but did not reverse it. Taken together, our data suggest that redox activation of RyRs by ROS has an important role in transducing the sustained contraction of pulmonary arteries under hypoxia. PMID- 16373825 TI - Molecular and cellular determinants of lung endothelial cell heterogeneity. AB - Idiopathic pulmonary arterial hypertension is a progressive and potentially fatal disease with a limited number of therapeutic options. Two key lesions underlie the pathophysiology of this disease. The principal lesion is found in large- and intermediate-sized blood vessels and is characterized by medial and adventitial hypertrophy/hyperplasia, with distal extension of smooth-muscle layers into normally nonmuscularized vessels. The second lesion, found prominently in severe forms of pulmonary hypertension, originates in small precapillary vessel segments, commonly at blood vessel bifurcations. This "plexiform lesion" is a lumen-obliterative lesion comprised, at least in part, of cells that share endothelial cell attributes, but that have lost the "law of the monolayer." Indeed, the endothelial contribution to the (mal-)adaptive response in pulmonary hypertension is becoming increasingly apparent, with evidence that endothelium plays an important role in promoting the vasoconstriction and hyperproliferation of medial and adventitial cell layers in large- and intermediate-vessel sizes, and lumen obliteration in the plexiform lesion. Recent evidence indicates endothelial cells along the pulmonary artery and precapillary segments are phenotypically distinct and may fulfill different roles in these site-specific lesions. Thus, the present review summarizes our current understanding of pulmonary endothelial cell heterogeneity and discusses the potential role(s) of endothelial cell heterogeneity in the pathogenesis of pulmonary hypertension. PMID- 16373826 TI - Aberrant signal transduction in pulmonary hypertension. PMID- 16373827 TI - The extracellular matrix microenvironment specifies pulmonary endothelial cell identity: roles of tenascin-C and RhoA. PMID- 16373828 TI - Pulmonary hypertension: cellular and molecular mechanisms. PMID- 16373829 TI - The protective role of T-lymphocytes in pulmonary vascular remodeling. PMID- 16373830 TI - Definitive evidence of fundamental and inherent alteration in the phenotype of primary pulmonary hypertension endothelial cells in angiogenesis. PMID- 16373831 TI - Interleukin-6 causes mild pulmonary hypertension and augments hypoxia-induced pulmonary hypertension in mice. PMID- 16373832 TI - Matrix metalloproteinases in the urine of patients with pulmonary arterial hypertension. PMID- 16373833 TI - The role of p38 mitogen-activated protein kinase in hypoxia-induced vascular cell proliferation: an interspecies comparison. PMID- 16373834 TI - Effects of simvastatin on cigarette smoking-induced structural and functional changes in rat lungs. PMID- 16373835 TI - Pulmonary capillary hemangiomatosis: results of gene expression analysis. PMID- 16373836 TI - The expression of prostacyclin synthase is decreased in the small pulmonary arteries from patients with emphysema. PMID- 16373837 TI - Protein kinase C inhibits cyclic adenosine monophosphate-induced calcium activated and voltage-activated potassium channel activity in fawn-hooded rat pulmonary arterial smooth muscle via phosphodiesterases. PMID- 16373838 TI - Pulmonary capillary hemangiomatosis: an immunohistochemical analysis of vascular remodeling in a fatal case. PMID- 16373839 TI - Hypoxia down-regulates cyclic guanidine monophosphate-dependent protein kinase in fetal pulmonary vascular smooth muscle cell through generation of reactive oxygen species and promotes development of pulmonary hypertension. PMID- 16373840 TI - Mts1/S100A4 stimulates human pulmonary artery smooth muscle cell migration through multiple signaling pathways. PMID- 16373841 TI - Mechanisms of heme oxygenase-1-mediated cardiac and pulmonary vascular protection in chronic hypoxia: roles of carbon monoxide and bilirubin. PMID- 16373842 TI - Differential gene expression in chronic hypoxic pulmonary hypertension: effect of simvastatin treatment. PMID- 16373843 TI - Endothelin-B receptor overexpression prevents hypoxic pulmonary hypertension in cirrhotic rats. PMID- 16373844 TI - Attenuation of hypoxia-induced pulmonary artery remodeling by a peroxisome proliferator-activated receptor-gamma agonist. PMID- 16373846 TI - Role of rho in the increased migration of pulmonary artery smooth muscle cells observed in the neprilysin knockout mouse. PMID- 16373845 TI - The low-voltage-activated calcium channel CAV3.1 controls proliferation of human pulmonary artery myocytes. PMID- 16373847 TI - Hypoxia-induced alterations in protein kinase C zeta signaling result in augmented fibroblast proliferation. PMID- 16373848 TI - Circulating mononuclear cells with a dual, macrophage-fibroblast phenotype contribute robustly to hypoxia-induced pulmonary adventitial remodeling. PMID- 16373849 TI - Microarray analysis of peripheral blood cells in pulmonary arterial hypertension, surrogate to biopsy. PMID- 16373850 TI - Transforming growth factor-beta signal transduction in angiogenesis and vascular disorders. AB - Transforming growth factor (TGF)-beta is a multifunctional protein that initiates its diverse cellular responses by binding to and activating specific type I and type II serine/threonine kinase receptors. TGF-beta can act as a regulator of proliferation, migration, survival, differentiation, and extracellular matrix synthesis in endothelial cells and vascular smooth muscle cells, as well as in the maintenance of vascular homeostasis. Importantly, genetic studies in humans have revealed the pivotal role of TGF-beta as well as its signaling components in angiogenesis. Mutations in two TGF-beta receptors (ie, the activin receptor-like kinase (ALK) 1 and the accessory TGF-beta receptor endoglin) have been linked to vascular disorders named hereditary hemorrhagic telangiectasia. In addition, knockout mice for the different components of the TGF-beta signaling pathway have shown that TGF-beta is indispensable for angiogenesis. Recent studies have revealed that TGF-beta can regulate vascular homeostasis by balancing the signaling between two distinct TGF-beta type I receptors (ie, the endothelial restricted ALK1 and the broadly expressed ALK5 receptors). The activation of these receptors has been shown to induce opposite effects on endothelial cell behavior and angiogenesis. In this review, we will present recent advances in understanding the role of TGF-beta signaling in endothelial cells as well as the underlying molecular mechanisms by which perturbation of this pathway can lead to vascular disorders. PMID- 16373851 TI - Bone morphogenetic protein 4 promotes vascular remodeling in hypoxic pulmonary hypertension. PMID- 16373852 TI - Regulation and functions of the paired-related homeobox gene PRX1 in pulmonary vascular development and disease. PMID- 16373853 TI - Involvement of hypoxia-inducible factor 1 in pulmonary pathophysiology. AB - Hypoxia-inducible factor (HIF)-1 is a transcription factor that is activated in response to hypoxia and growth factor/cytokine signaling via regulation of the HIF-1alpha subunit. HIF-1 has been implicated in the pathogenesis of pulmonary hypertension based on both experimental and clinical data. In a mouse model of pulmonary hypertension, hypoxia-induced increases in right ventricular mass, right ventricular pressure, and medial wall thickness of pulmonary arterioles were impaired in mice that were heterozygous for a null allele at the locus encoding HIF-1alpha compared to wild-type littermates. Electrophysiologic analyses revealed that the hypoxia-induced hypertrophy and depolarization of pulmonary arterial smooth muscle cells from wild-type mice was significantly impaired in heterozygotes. In clinical studies, immunohistochemical analyses of plexiform lesions within the lungs of patients with severe pulmonary hypertension revealed dramatic overexpression of HIF-1alpha within proliferating endothelial cells. These cells also expressed vascular endothelial growth factor (VEGF), which is the product of a known HIF-1 target gene, indicating that autocrine VEGF VEGF receptor signaling may contribution to the pathogenesis of plexiform lesions. These studies implicate HIF-1 in pathophysiologic alterations of both smooth muscle and endothelial cell biology in patients with pulmonary hypertension. PMID- 16373854 TI - Chronic hypoxia-enhanced murine pulmonary vasoconstriction: role of superoxide and gp91phox. AB - Chronic hypoxia (CH) is a common cause of pulmonary hypertension (PH). Accumulating evidence suggests that changes in the activity of endothelin (ET)-1 receptors may play an important role in CH-induced PH. After 3 weeks of CH (10% O2) exposure, we found that the isolated intra-pulmonary artery (PA) constrictor response to ET-1 was significantly increased in wild-type (wt) mice. The administration of Cu/Zn superoxide dismutase (SOD) markedly reduced the CH enhanced maximal PA constrictor response to ET-1, demonstrating the contribution of superoxide to CH-enhanced PA constrictor responses. Using mice that are completely deficient in gp91phox (a subunit protein of the superoxide producing nicotinamide adenine dinucleotide phosphate [NADPH] oxidase), we found that CH enhanced PA constriction to ET-1 was completely blocked (decreases in mean [+/- SE] maximal isometric tension from 5.43 +/- 0.35 to 3.33 +/- 0.19 mN; n = 7; p < 0.01). Using a lucigenin-enhanced chemiluminescence technique to measure superoxide, we found that the 3 weeks of CH significantly increased superoxide levels in PA isolated from wt mice. The addition of ET-1 further increased superoxide production. To demonstrate that the increased chemiluminescence is due to superoxide generation, we added Cu/Zn SOD, which markedly decreased chemiluminescence, demonstrating the specificity of this assay. When gp91phox knockout mice were exposed to CH, they had significantly reduced levels of superoxide compared to CH-treated wt mice. Our results demonstrate that the CH enhanced PA constrictor response to ET-1 is mediated by NADPH oxidase (gp91phox) derived superoxide overproduction that may contribute to the pathogenesis of CH induced PH. PMID- 16373855 TI - Phenotypic diversity of the lung vasculature in experimental models of metastases. AB - In vivo phage display is a screening method in which peptides homing to specific vascular beds are selected after IV administration of a random peptide library. This strategy has revealed a vascular address system that allows tissue-specific targeting of normal blood vessels and angiogenesis-related targeting of tumor blood vessels by selected peptides. Many vascular receptors or "addresses" targeted by homing peptides have been identified. One such vascular receptor of normal lung endothelium is membrane dipeptidase (MDP), which was found by in vivo phage display to bind the tripeptide motif gly-phe-glu (GFE). Our studies with GFE peptide and lung vasculature suggest that MDP mediates cancer cell adhesion to lung vasculature and the development of lung metastases, but that MDP is not present in the vasculature of lung metastases. MDP appears to occupy a vascular distribution that is similar to the pulmonary artery circulation. These results demonstrate the promise of defining critical functional and anatomic characteristics of endothelial cells in lung and other organs by in vivo phage display. PMID- 16373856 TI - Hypoxia amplifies the proliferative capacity of distal human pulmonary artery smooth-muscle cells. PMID- 16373857 TI - Abnormal vascular phenotypes in patients with idiopathic pulmonary fibrosis and secondary pulmonary hypertension. PMID- 16373858 TI - Imaging of angiogenesis in inflamed airways and tumors: newly formed blood vessels are not alike and may be wildly abnormal: Parker B. Francis lecture. PMID- 16373859 TI - Does BMPR2 mutation disrupt pulmonary vasculogenesis? PMID- 16373860 TI - Extracellular adenosine triphosphate: a potential regulator of vasa vasorum neovascularization in hypoxia-induced pulmonary vascular remodeling. PMID- 16373861 TI - Vascular endothelial growth factor receptor blockade by SU5416 combined with pulsatile shear stress causes apoptosis and subsequent proliferation of apoptosis resistant endothelial cells. PMID- 16373862 TI - Rho guanosine triphosphatases in lung development and pulmonary hypertension. PMID- 16373863 TI - Genomic threats from physiological signals in lung vascular cells. PMID- 16373864 TI - Tissue factor is induced in a rodent model of severe pulmonary hypertension characterized by neointimal lesions typical of human disease. PMID- 16373865 TI - Prostacyclin synthase promoter regulation and familial pulmonary arterial hypertension. PMID- 16373866 TI - Inhaled nitric oxide reverses hypoxia induced lung hypoplasia in endothelial nitric oxide synthase-deficient mice. PMID- 16373867 TI - Vascular endothelial growth factor improves pulmonary vascular reactivity and structure in an experimental model of chronic pulmonary hypertension in fetal sheep. PMID- 16373868 TI - Ventilation/perfusion ratios in pulmonary arterial hypertension: effects of IV and inhaled prostacyclin derivatives. PMID- 16373869 TI - Comparative analysis of BMPR2 gene and its mutations in idiopathic pulmonary arterial hypertension. PMID- 16373870 TI - The impact of pulmonary hypertension on survival in patients with idiopathic pulmonary fibrosis. PMID- 16373871 TI - Effects of aerosol vs IV UT-15 on prostaglandin H2 analog-induced pulmonary hypertension in sheep. PMID- 16373872 TI - Evidence for systemic endothelial dysfunction in patients and first-order relatives with pulmonary arterial hypertension. PMID- 16373873 TI - Acute cardiopulmonary hemodynamic effects of brain natriuretic peptide in patients with pulmonary arterial hypertension. PMID- 16373874 TI - Bone morphogenetic protein receptor 2 mutations in adults and children with idiopathic pulmonary arterial hypertension: association with thyroid disease. PMID- 16373875 TI - Metastatic cancer while receiving continuous prostacyclin therapy. PMID- 16373876 TI - Serotonin transporter gene polymorphism in a cohort of German patients with idiopathic pulmonary arterial hypertension or chronic thromboembolic pulmonary hypertension. PMID- 16373877 TI - Divergent contractile and structural responses of the murine protein kinase C epsilon null pulmonary circulation to chronic hypoxia. PMID- 16373878 TI - Evidence for vascular remodeling in the lungs of macaques infected with simian immunodeficiency virus/HIV NEF recombinant virus. PMID- 16373879 TI - Osteopontin may be responsible for pulmonary vascular remodeling. PMID- 16373880 TI - Clinical challenges in pulmonary hypertension: Roger S. Mitchell lecture. AB - Despite major advances in our understanding of the pathophysiologic processes leading to pulmonary arterial hypertension and recent developments in therapeutic approaches, the long-term prognosis for patients with pulmonary arterial hypertension remains unsatisfactory. Early detection and adequate clinical classification of the disease, better assessment of patients' prognosis, and improved therapeutic strategies are important challenges for clinicians in coming years. PMID- 16373881 TI - Pulmonary endothelin-1 clearance in human pulmonary arterial hypertension. PMID- 16373882 TI - Interventricular mechanical asynchrony due to right ventricular pressure overload in pulmonary hypertension plays an important role in impaired left ventricular filling. PMID- 16373883 TI - Enhanced hypoxic pulmonary vasoconstriction in families of adults or children with idiopathic pulmonary arterial hypertension. PMID- 16373884 TI - MRI pulmonary angiograms. PMID- 16373885 TI - Role of angiopoietin-1 in experimental and human pulmonary arterial hypertension. AB - INTRODUCTION: The pulmonary microvasculature, consisting mainly of an endothelial cell (EC) monolayer and scant matrix support, is incompletely muscularized. Thus, the distal pulmonary arterioles may be predisposed to regression on exposure to environmental stresses (ie, hypoxia) and may be dependent on EC survival factors, like angiopoietin (Ang) 1, to attenuate the development of pulmonary arterial hypertension (PAH). In order to clarify the link between Ang1 expression and the development of PAH in patients, we also studied messenger RNA and protein expression in lung samples from healthy control subjects and patients with idiopathic PAH (IPAH) or PAH associated with other diseases (APAH). METHODS: Ang/Tie2 gene expression was assessed in rats that had been exposed to hypoxia (ie, 10% O2) for 1, 3, or 7 days. In a separate experiment, the cell-based gene transfer of Ang1/Ang2 was performed, and the effects were evaluated in rats with hypoxia-induced PAH. RESULTS: Hypoxia induced significant early increases in right ventricular systolic pressure (RVSP) and right ventricle/left ventricle plus-septum mass ratio (RV/[LV + S]), with a significant decrease in Tie2 expression. Hypoxic rats receiving Ang1 demonstrated significant improvements in RVSP and RV/(LV + S), with a partial normalization in Tie2 protein levels. Robust Ang1 expression was observed in healthy human lungs. Furthermore, there were no significant changes in the levels of Ang1 or Ang2 in IPAH or APAH samples vs those in control subjects. CONCLUSIONS: Decreased activity of the Tie2 pathway with hypoxia may contribute to PAH, possibly by loss of EC survival signaling, which can be overcome by Ang1 gene transfer. PMID- 16373886 TI - Cellular and molecular pathobiology of pulmonary hypertension conference summary. PMID- 16373887 TI - Impact of active versus usual algorithmic titration of basal insulin and point-of care versus laboratory measurement of HbA1c on glycemic control in patients with type 2 diabetes: the Glycemic Optimization with Algorithms and Labs at Point of Care (GOAL A1C) trial. AB - OBJECTIVE: The objective of this study was to assess the impact of active versus usual monitoring of algorithmic insulin titration and point-of-care (POC) versus laboratory HbA1c (A1C) measurement on glycemic control in primary care. RESEARCH DESIGN AND METHODS: The Glycemic Optimization with Algorithms and Labs at Point of Care (GOAL A1C) study was a 24-week, randomized, parallel-group, four-arm, open-label study of 7,893 adults with type 2 diabetes uncontrolled by oral antidiabetic agents and requiring insulin. Patients were randomly assigned by investigators from 2,164 sites in the U.S. to insulin glargine with either 1) usual (no unsolicited contact between visits) insulin titration using a simple algorithm with laboratory A1C testing, 2) usual titration with POC A1C testing, 3) active (weekly monitored) titration with laboratory A1C testing, or 4) active titration with POC A1C testing. Outcome measures included a change in A1C and fasting self-monitoring of blood glucose (SMBG) levels, percentage of patients achieving A1C <7.0%, and hypoglycemia frequency. RESULTS: Significant A1C and SMBG reductions were observed in all arms (P < 0.0001). Compared with usual insulin titration, active titration achieved greater A1C reduction (1.5 vs. 1.3%; P < 0.0001), SMBG reduction (88 vs. 79 mg/dl; P < 0.0001), and proportion of patients achieving A1C <7.0% (38 vs. 30%; P < 0.0001). Among patients receiving active titration, POC A1C testing was associated with an increase in the proportion achieving an A1C <7.0% (41% for POC vs. 36% for laboratory; P < 0.0001). Hypoglycemia rates were low (usual vs. active groups: 3.7 vs. 6.0 all confirmed episodes/patient-year [P < 0.001]; 0.09 vs. 0.14 severe episodes/patient-year [NS]). CONCLUSIONS: In a predominantly primary care setting, addition of insulin glargine using a simple algorithm achieved significant improvements in glycemic control in patients with type 2 diabetes in all four study arms. Active titration resulted in significant incremental improvements in glycemic control, and, among patients receiving active titration, POC A1C testing resulted in a greater portion achieving A1C <7.0%. PMID- 16373888 TI - Treatment disparities in the care of patients with and without diabetes presenting with non-ST-segment elevation acute coronary syndromes. AB - OBJECTIVE: The objective of this study was to characterize treatment patterns among patients with diabetes presenting with non-ST-segment elevation (NSTE) acute coronary syndromes (ACSs). RESEARCH DESIGN AND METHODS: We compared adherence to treatment recommendations from the American College of Cardiology (ACC)/American Heart Association (AHA) guidelines for NSTE ACS among 46,410 patients from 413 U.S. hospitals that were included in the Can Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes with Early Implementation of the ACC/AHA Guidelines (CRUSADE) quality improvement initiative. Patients were stratified as nondiabetic, non-insulin-dependent diabetic (type 2 diabetic), and insulin-treated diabetic. RESULTS: Insulin treated diabetic patients were less likely than nondiabetic patients to receive aspirin (adjusted odds ratio 0.83 [95% CI 0.74-0.93]), beta-blockers (0.89 [0.83 0.96]), heparin (0.90 [0.83-0.98]), and glycoprotein IIb/IIIa inhibitors (0.86 [0.79-0.93]). Type 2 diabetic patients were treated similarly to nondiabetic patients. After adjustment for differences in clinical characteristics, insulin treated diabetic patients were significantly less likely than nondiabetic patients to receive cardiac catheterization within 48 h of presentation (0.80 [0.74-0.86]) or percutaneous coronary intervention (0.87 [0.82-0.94]). Compared with nondiabetic patients, insulin-treated diabetic and type 2 diabetic patients were more likely to undergo coronary artery bypass grafting (1.34 [1.21-1.49] and 1.35 [1.26-1.44]). In-hospital mortality rates were higher in insulin-treated diabetic (6.8%) and type 2 diabetic (5.4%) than in nondiabetic (4.4%) patients. CONCLUSIONS: Diabetic patients have a higher risk of mortality than nondiabetic patients, yet physicians adhere to the ACC/AHA NSTE ACS guidelines less often when treating diabetic patients, particularly insulin-treated diabetic patients. Increased use of guideline-recommended therapies and early invasive management strategies in diabetic patients may improve their outcomes. PMID- 16373889 TI - Association between complementary and alternative medicine use, preventive care practices, and use of conventional medical services among adults with diabetes. AB - OBJECTIVE: To assess the association between complementary and alternative medicine (CAM) use, preventive care practices, and use of conventional medical services among adults with diabetes. RESEARCH DESIGN AND METHODS: We analyzed data on 2,474 adults with diabetes. We created an overall CAM-use category based on use of any of the following: diets, herbs, chiropractic care, yoga, relaxation, acupuncture, ayuverda, biofeedback, chelation, energy healing, Reiki therapy, hypnosis, massage, naturopathy, and homeopathy. We used multiple logistic regression to assess the effect of CAM use on preventive care practices (receipt of influenza and pneumonia vaccines) and use of conventional medical services (number of primary care and emergency department visits). STATA was used for statistical analysis to account for the complex survey design. RESULTS: A total of 48% of adults with diabetes used some form of CAM. CAM use was independently associated with receipt of pneumonia vaccination (odds ratio 1.56 [95% CI 1.26-1.94]) but not significantly associated with receipt of influenza vaccination (1.17 [0.92-1.48]). CAM use was independently associated with visiting the emergency department (1.34 [1.06-1.70]), having six or more primary care visits (1.44 [1.14-1.83]), and having eight or more primary care visits (1.66 [1.22-2.25]). CONCLUSIONS: In contrast to the findings of previous studies, CAM use appears to be associated with increased likelihood of receipt of preventive care services and increased emergency department and primary care visits. CAM use may not be a barrier to use of conventional medical services in adults with diabetes. PMID- 16373890 TI - The effects of aerobic exercise on glucose and counterregulatory hormone concentrations in children with type 1 diabetes. AB - OBJECTIVE: To examine the acute glucose-lowering effects of aerobic exercise in children and adolescents with type 1 diabetes. RESEARCH DESIGN AND METHODS: Fifty children and adolescents with type 1 diabetes (ages 10 to <18 years) were studied during exercise. The 75-min exercise session consisted of four 15-min periods of walking on a treadmill to a target heart rate of 140 bpm and three 5-min rest periods. Blood glucose and plasma glucagon, cortisol, growth hormone, and norepinephrine concentrations were measured before, during, and after exercise. RESULTS: In most subjects (83%), plasma glucose concentration dropped at least 25% from baseline, and 15 (30%) subjects became hypoglycemic (< or = 60 mg/dl) or were treated for low glucose either during or immediately following the exercise session. The incidence of hypoglycemia and/or treatment for low glucose varied significantly by baseline glucose, occurring in 86 vs. 13 vs. 6% of subjects with baseline values <120, 120-180, and >180 mg/dl, respectively (P < 0.001). Exercise induced increases in growth hormone and norepinephrine concentrations were marginally higher in subjects whose glucose dropped < or = 70 mg/dl. Treatment of hypoglycemia with 15 g of oral glucose resulted in only about a 20-mg/dl rise in glucose concentrations. CONCLUSIONS: In youth with type 1 diabetes, prolonged moderate aerobic exercise results in a consistent reduction in plasma glucose and the frequent occurrence of hypoglycemia when preexercise glucose concentrations are <120 mg/dl. Moreover, treatment with 15 g of oral glucose is often insufficient to reliably treat hypoglycemia during exercise in these youngsters. PMID- 16373891 TI - Relation between blood glucose and coronary mortality over 33 years in the Whitehall Study. AB - OBJECTIVE: Glucose intolerance is a risk factor for coronary disease, but there is uncertainty about the shape of the dose-response relationship between glucose level and risk of coronary mortality. We examined the prospective relation of 2-h postload blood glucose (2hBG) with coronary and other major causes of mortality over 33 years. RESEARCH DESIGN AND METHODS: A 50-g oral glucose tolerance test (OGTT) was performed at baseline (1967-1969) in 17,869 male civil servants aged 40-64 years. RESULTS: There were 3,561 coronary deaths during 451,787 person years of observation. All-cause, cardiovascular, and respiratory mortality were elevated among participants with glucose intolerance. The hazard of coronary mortality rose from 2hBG = 4.6 mmol/l (83 mg/dl [95% CI 4.2-5.3]). The dose response relation was best fitted by a single slope above this level, with no evidence of nonlinearity, compared with Cox models using other threshold levels, and those containing log 2hBG terms. There was no evidence for a dose-response relationship below 2hBG = 4.6 mmol/l. Between this level and 11.1 mmol/l (200 mg/dl), the age-adjusted hazard ratio was 3.62 (95% CI 2.3-5.6). The graded relationship was attenuated by 45% after adjustment for baseline coronary heart disease (CHD), BMI, systolic blood pressure, blood cholesterol, smoking, physical activity, lung function, and employment grade. CONCLUSIONS: A threshold model with linear slope best described the dose-response relationship between postload blood glucose and CHD mortality risk. PMID- 16373892 TI - Recent trends in cardiovascular complications among men and women with and without diabetes. AB - OBJECTIVE: To compare recent trends in cardiovascular disease (CVD) outcomes among men and women with diabetes with those in the nondiabetic population. RESEARCH DESIGN AND METHODS: We conducted a retrospective cohort study using provincial health claims data to identify adults with (n = 670,602) and without (n = 9,190,721) diabetes living in Ontario, Canada, between 1 April 1992 and 31 March 2000. We compared changes in the annual age-/sex-adjusted rates and numbers of subjects admitted for acute myocardial infarction (AMI) and stroke and of deaths from AMI, stroke, and all causes between those with and without diabetes. RESULTS: Over the 8-year period, the rate of patients admitted for AMI and stroke fell to a greater extent in the diabetic than the nondiabetic population (AMI: 15.1 vs. -9.1%, P < 0.0001; stroke: -24.2 vs. 19.4%, P < 0.0001). Diabetic patients experienced similar reductions in case-fatality rates related to AMI and stroke than those without diabetes (-44.1 vs. -33.2%, P = 0.1; -17.1 vs. -16.6%, P = 0.9, respectively). Declines in all-cause mortality were also comparable in the two populations. Over the same period, the number of diabetes cases increased from 405,471 to 670,602. Thus, while CVD rates fell, the number of events occurring in this population rose substantially (AMI: +44.6%, stroke: +26.1%, AMI deaths: +17.2%, and stroke deaths: +13.2%). CONCLUSIONS: Our findings demonstrate a significant reduction in the rate of people affected by CVD within the diabetic population. However, as the number of people with diabetes rises, so may the absolute burden of CVD in our society. PMID- 16373893 TI - Physical activity and life expectancy with and without diabetes: life table analysis of the Framingham Heart Study. AB - OBJECTIVE: Physical activity is associated with a reduced risk of developing diabetes and with reduced mortality among diabetic patients. However, the effects of physical activity on the number of years lived with and without diabetes are unclear. Our aim is to calculate the differences in life expectancy with and without type 2 diabetes associated with different levels of physical activity. RESEARCH DESIGN AND METHODS: Using data from the Framingham Heart Study, we constructed multistate life tables starting at age 50 years for men and women. Transition rates by level of physical activity were derived for three transitions: nondiabetic to death, nondiabetic to diabetes, and diabetes to death. We used hazard ratios associated with different physical activity levels after adjustment for age, sex, and potential confounders. RESULTS: For men and women with moderate physical activity, life expectancy without diabetes at age 50 years was 2.3 (95% CI 1.2-3.4) years longer than for subjects in the low physical activity group. For men and women with high physical activity, these differences were 4.2 (2.9-5.5) and 4.0 (2.8-5.1) years, respectively. Life expectancy with diabetes was 0.5 (-1.0 to 0.0) and 0.6 (-1.1 to -0.1) years less for moderately active men and women compared with their sedentary counterparts. For high activity, these differences were 0.1 (-0.7 to 0.5) and 0.2 (-0.8 to 0.3) years, respectively. CONCLUSIONS: Moderately and highly active people have a longer total life expectancy and live more years free of diabetes than their sedentary counterparts but do not spend more years with diabetes. PMID- 16373894 TI - Improvement in glycemic excursions with a transcutaneous, real-time continuous glucose sensor: a randomized controlled trial. AB - OBJECTIVE: Hypoglycemia and wide glucose excursions continue to be major obstacles to achieving target HbA(1c) values and the associated reductions in long-term complications (and economic costs) in people with insulin-treated diabetes. In this study we evaluated the accuracy, safety, and clinical effectiveness of a continuous glucose-sensing device. RESEARCH DESIGN AND METHODS: A total of 91 insulin-requiring patients with type 1 (n = 75) and type 2 (n = 16) diabetes were enrolled in this multicenter randomized study. Subjects wore a transcutaneous, 3-day, continuous glucose-sensing system for three consecutive 72-h periods. Subjects were randomly assigned (1:1 ratio) to either a control group (continuous glucose data not provided) or a display group (continuous glucose data not provided during period 1 but displayed during periods 2 and 3). During periods 2 and 3, patients in the display group had real time access to sensor glucose values, could review glucose trends over the preceding 1, 3, and 9 h, and were provided with high (> or = 200 mg/dl) and low (< or = 80 mg/dl) alerts and a low (< or = 55 mg/dl) alarm. Sensors were inserted by patients, and both groups used (or wore) the system during daily activities. Device accuracy was assessed by comparing continuous glucose values to paired self-monitoring of blood glucose (SMBG) meter readings. Clinical effectiveness was evaluated by analyzing between-group (control vs. display, periods 2 and 3) and within-group (display, period 1 vs. period 3) differences in time spent in high, low, and target (81-140 mg/dl) glucose zones. RESULTS: When prospective, real-time sensor values were compared with SMBG values, 95.4% of 6,767 paired glucose values fell within Clarke error grid A and B zones. Pearson's correlation coefficient was 0.88, and mean and median absolute relative differences were 21.2 and 15.9%, respectively. No systematic bias was detected at any of the prespecified glucose levels (50, 80, 100, 150, and 200 mg/dl). When compared with control subjects, the display group spent 21% less time as hypoglycemic (<55 mg/dl), 23% less time as hyperglycemic (> or = 240 mg/dl), and 26% more time in the target (81-140 mg/dl) glucose range (P < 0.001 for each comparison). Nocturnal (10:00 p.m. to 6:00 a.m.) hypoglycemia, as assessed at two thresholds, was also reduced by 38% (<55 mg/dl; P < 0.001) and 33% (55-80 mg/dl; P < 0.001) in the display group compared with control subjects. CONCLUSIONS: We conclude that real-time continuous glucose monitoring for periods up to 72 h is accurate and safe in insulin-requiring subjects with type 1 and type 2 diabetes. This study demonstrates that availability of real-time, continuously measured glucose levels can significantly improve glycemic excursions by reducing exposure to hyperglycemia without increasing the risk of hypoglycemia, which may reduce long term diabetes complications and their associated economic costs. PMID- 16373895 TI - Racial differences in adiponectin in youth: relationship to visceral fat and insulin sensitivity. AB - OBJECTIVE: The purpose of this study was to investigate 1) whether adiponectin is associated with insulin sensitivity independent of visceral adipose tissue in African-American and Caucasian youth and 2) whether adiponectin is associated with racial differences in insulin sensitivity. RESEARCH DESIGN AND METHODS: Total body fat was measured by dual-energy X-ray absorptiometry and abdominal adipose tissue with computed tomography. Insulin sensitivity was measured by a 3 h hyperinsulinemic-euglycemic clamp. RESULTS: Adiponectin was inversely associated (P < 0.01) with visceral adipose tissue, fasting insulin, and proinsulin and was positively related (P < 0.01) to insulin sensitivity after controlling for Tanner stage and sex independent of race. Stepwise multiple regression revealed that adiponectin was a strong independent predictor of insulin sensitivity, explaining 27% of the variance in insulin sensitivity. When subjects were categorized into tertiles of visceral adipose tissue and further low (< or = 50th) and high (>50th) adiponectin groups, insulin sensitivity was significantly different across the visceral adipose tissue groups (main effect, P < 0.01) in both races. However, within each visceral adipose tissue group, subjects with high adiponectin had higher insulin sensitivity (main effect, P < 0.05) than subjects with low adiponectin, independent of race. Racial differences in insulin sensitivity remained significant (P < 0.01) after controlling for leptin and visceral adipose tissue but not (P > 0.05) after additional adjustment for adiponectin. CONCLUSIONS: Adiponectin is associated with insulin sensitivity independent of visceral adipose tissue in both African-American and Caucasian youth. Low adiponectin in African-American youth may be a biological marker that predisposes them to a greater risk of insulin resistance. PMID- 16373896 TI - Impaired glucose tolerance of pregnancy is a heterogeneous metabolic disorder as defined by the glycemic response to the oral glucose tolerance test. AB - OBJECTIVE: Gestational diabetes mellitus (GDM), defined by two abnormal glucose values on a 3-h oral glucose tolerance test (OGTT), is associated with insulin resistance and a low serum concentration of adiponectin. The metabolic implications of impaired glucose tolerance (IGT) of pregnancy (i.e., a single abnormal value on an OGTT), however, are not well established. We sought to evaluate the metabolic phenotype of pregnant women with IGT in relation to the timing of their isolated hyperglycemia. RESEARCH DESIGN AND METHODS: A cross sectional study was performed in pregnant women undergoing a 3-h, 100-g OGTT. The OGTT stratified participants into four groups: 1) GDM (n = 48), 2) 1-h IGT (single elevated value at 1 h) (n = 15), 3) 2-h/3-h IGT (single elevated value at either 2 or 3 h) (n = 23), and 4) normal glucose tolerance (NGT) (n = 93). Insulin sensitivity was measured by the validated insulin sensitivity index (IS(OGTT)) of Matsuda and DeFronzo. RESULTS: Measures of severity of glycemia (fasting glucose, area under the glucose curve from the OGTT, and glucose challenge test result) were highest in the GDM group, followed by the 1-h IGT, 2 h/3-h IGT, and NGT groups, respectively (each trend P < 0.0001). Consistent with this finding, IS(OGTT) was highest in the NGT group (5.1), followed by the 2-h/3 h IGT (4.6), 1-h IGT (3.8), and GDM (3.2) groups (trend P < 0.0001). Furthermore, on multiple linear regression analysis of IS(OGTT), both GDM and 1-h IGT were independently associated with reduced insulin sensitivity (whereas 2-h/3-h IGT was not). Mean adjusted adiponectin was highest in the NGT group (15.7 microg/ml), followed by the 2-h/3-h IGT (15.6 microg/ml), 1-h IGT (13.7 microg/ml), and GDM (12.0 microg/ml) groups (trend P = 0.0024). CONCLUSIONS: The metabolic implications of IGT in pregnancy vary in relation to the timing of the abnormal glucose value from the diagnostic OGTT. The metabolic phenotype associated with 1-h IGT resembles that of GDM, whereas the phenotype associated with 2-h/3-h IGT exhibits similarity to that of NGT. PMID- 16373897 TI - The fidgety fetus hypothesis: fetal activity is an additional variable in determining birth weight of offspring of women with diabetes. AB - OBJECTIVE: To determine whether some offspring of women with diabetes are intrinsically more active than others in utero and whether those who are active are able to normalize their birth weight despite maternal hyperglycemia. RESEARCH DESIGN AND METHODS: We conducted a three-phase study to view the relationship between fetal movements and subsequent birth weight in women with diabetes. Phase I was designed to assess maternal perception of fetal movements in a population of 10 women with diabetes. To improve our fetal monitoring techniques, in phase II we analyzed fetal movements using the Card Guard home fetal monitoring device (CG 900P) in a population of 13 women with gestational diabetes mellitus (GDM). To apply our observations of fetal movements to a larger population, during phase III we conducted a retrospective analysis of fetal monitoring strips (HP 8041A) from 46 women with GDM to examine the relationship between fetal heart rate (FHR) accelerations and percentile birth weight, corrected for gestational age. RESULTS: Phase I confirmed that there is little variability in fetal movements (i.e., fetal kicks did not significantly deviate from one another on a day-to-day basis). In phase II, the fetal monitoring strips illustrated that the active fetuses (defined as > or = 4 FHR accelerations in a 20-min period) were always active, and the inactive fetuses were always inactive. The mean birth weight percentile, corrected for gestational age, in the active group was 37 vs. 63% in the inactive group (P = 0.05). In phase III, the fetal monitoring strips showed an inverse correlation between the mean number of FHR accelerations and the birth weight of the fetus, corrected for gestational age. The mean birth weight percentile in the active group was 37 vs. 62% in the inactive group (P = 0.0017). CONCLUSIONS: The fetus appears to play a role in determining its own destiny. Increased fetal activity may minimize the impact of hyperglycemia on subsequent birth weight. The inactive fetus appears to be at a higher risk for glucose mediated macrosomia. PMID- 16373898 TI - Long-term effects of ranirestat (AS-3201) on peripheral nerve function in patients with diabetic sensorimotor polyneuropathy. AB - OBJECTIVES: We aimed to determine whether ranirestat, an aldose reductase inhibitor, maintains the improved nerve function observed in patients with diabetic sensorimotor polyneuropathy (DSP) after completing a 12-week nerve biopsy study. RESEARCH DESIGN AND METHODS: Patients with mild to moderate DSP, as determined by the presence of sural nerve responses, were enrolled in a double blind, placebo-controlled biopsy trial and randomized to placebo or 5 or 20 mg/day ranirestat for 12 weeks. Patients completing this biopsy study were offered a 48-week extension at the same ranirestat dose or at 5 mg/day ranirestat if they were originally treated with placebo. Electrophysiological tests, the Toronto Clinical Neuropathy Score, and vibration perception thresholds (VPTs) were performed at entry and at 12 (end of the biopsy study) and 60 (end of the 48 week extension) weeks. RESULTS: Peroneal motor nerve conduction velocity (NCV) improved in the 20-mg/day group following 60 weeks of treatment. Sural and median sensory NCV improved after both 12 and 60 weeks of treatment with 20 mg/day. VPT improved after 60 weeks of treatment with 20 mg/day. Ranirestat was well tolerated with no difference in adverse events between the 5- and 20-mg/day groups. CONCLUSIONS: Twenty milligrams ranirestat per day improves NCV and VPT following 60 weeks of administration. The improved sensory nerve function observed after 12 weeks of therapy was maintained at 60 weeks, and improved motor nerve function was observed at 60 weeks. PMID- 16373899 TI - A low brain serotonergic neurotransmission in children with type 1 diabetes detected through the intensity dependence of auditory-evoked potentials. AB - OBJECTIVE: To determine in children with type 1 diabetes the plasma free fraction of L-tryptophan (FFT) and the intensity-dependent auditory-evoked potentials (IDAEPs) as indicators of possible changes in brain serotonergic neurotransmission. RESEARCH DESIGN AND METHODS: A prospective and comparative study was performed in children with type 1 diabetes and normal control subjects. We measured FFT, bound and total plasma L-tryptophan, neutral amino acids (NAAs), albumin, free fatty acids (FFAs), glucose, and HbA1c(A1C) and recorded IDAEPs with four intensities (40, 60, 90, and 103 dB). RESULTS: The glycemia, A1C, FFAs, and NAAs in plasma were significantly elevated. The FFT and the FFT-to-total L tryptophan and FFT-to-NAA ratios were reduced. The latencies of N1 and P2 increased at all intensities and the slope of the amplitude/stimulus intensity function (ASF slope) of the N1/P2 component significantly increased. CONCLUSIONS: The decrease of the FFT in plasma and increase in the N1/P2 component amplitude may reflect a functional relationship between the brain serotonergic activity with the N1/P2 changes. The increase of the ASF slope in children with type 1 diabetes suggests that the response of the auditory cortex to sound intensity stimulus may be regulated by the serotonergic tone and that decreased serotonergic neurotransmission may provoke a different behavior of sensory cortices. Therefore, the IDAEP (N1/P2 component) may be an electrophysiological indicator of brain changes of serotonergic neurotransmission in children with type 1 diabetes. These changes may be related to psychoemotional manifestations observed in diabetic children such as anxiety and depression. PMID- 16373900 TI - Higher levels of HDL cholesterol are associated with a decreased likelihood of albuminuria in patients with long-standing type 1 diabetes. AB - OBJECTIVE: The objective of this study was to determine whether high levels of HDL cholesterol are associated with a lower prevalence of albuminuria RESEARCH DESIGN AND METHODS: We analyzed the lipid profiles of patients with type 1 diabetes of > or = 20 years duration in 42 patients with albuminuria (28 microalbuminuria and 14 macroalbuminuria) and 65 patients without increased albumin excretion before any interventions with either statins or ACE inhibitors. RESULTS: Several characteristics were similar in the two groups: sex, age, duration of diabetes, total cholesterol, LDL cholesterol, and triglycerides. By univariate analysis, significant differences (P < 0.01) were found in HDL cholesterol (albuminuria 1.42 mg/dl, no albuminuria 1.71 mg/dl, P < 0.01), HbA1c (A1C) (albuminuria 8.5%, no albuminuria 7.5%), and proportions with no, background, and proliferative retinopathy (albuminuria 2.4, 16.7, and 81%; no albuminuria 24.6, 52.3, and 23.1%, respectively). When adjusted for age and sex, a 0.26-mmol/l (10-mg/dl) increase in HDL cholesterol is associated with an odds ratio (OR) of 0.70 (95% CI 0.54-0.90) for having albuminuria. In a multivariate model that adjusted for age, sex, diabetes duration, and A1C, for every 0.54 mmol/l (21-mg/dl) increase in HDL cholesterol, patients are approximately half (OR 0.51 [95% CI 0.30-0.86]) as likely to have albuminuria, even after controlling for A1C. CONCLUSIONS: Higher HDL cholesterol levels may be protective against the development of albuminuria in patients with type 1 diabetes. Whether this is due to the HDL cholesterol levels or whether they serve as a marker for some other mechanism remains to be determined. PMID- 16373901 TI - Urinary connective tissue growth factor excretion correlates with clinical markers of renal disease in a large population of type 1 diabetic patients with diabetic nephropathy. AB - OBJECTIVE: Levels of connective tissue growth factor (CTGF; CCN-2) in plasma are increased in various fibrotic disorders, including diabetic nephropathy. Recently, several articles have reported a strong increase of urinary CTGF excretion (U-CTGF) in patients with diabetic nephropathy. However, these studies addressed too small a number of patients to allow general conclusions to be drawn. Therefore, we evaluated U-CTGF in a large cross-sectional study of patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: Subjects were 318 type 1 diabetic patients and 29 normoglycemic control subjects. U-CTGF was measured by sandwich enzyme-linked immunosorbent assay. Groups were compared by Kruskal-Wallis and Mann-Whitney analysis. The relation between U-CTGF and markers of diabetic nephropathy was determined by regression analysis. RESULTS: U-CTGF in patients with diabetic nephropathy (n = 89, median 155 pmol/24 h [interquartile range 96-258]) was significantly higher than in microalbuminuric (n = 79, 100 [65 78]) and normoalbuminuric (n = 150, 85 [48-127]) patients and control subjects (n = 29, 100 [78-114]). U-CTGF correlated with urinary albumin excretion (UAE) (R = 0.31) and glomerular filtration rate (R = -0.38) in patients with diabetic nephropathy. A standardized increase in U-CTGF was associated with diabetic nephropathy (odds ratio 2.3 [95% CI 1.7-3.1]), which was comparable with the odds ratios for diabetic nephropathy of increased HbA(1c) (2.0 [1.5-2.7]), and blood pressure (2.0 [1.5-2.6]). CONCLUSIONS: This is the first large cross-sectional study addressing U-CTGF in human type 1 diabetes. The observed association of U CTGF with UAE and glomerular filtration rate might reflect a role of CTGF as progression promoter in diabetic nephropathy. PMID- 16373902 TI - Functional polymorphisms of UCP2 and UCP3 are associated with a reduced prevalence of diabetic neuropathy in patients with type 1 diabetes. AB - OBJECTIVE: We studied the association between polymorphisms in the UCP genes and diabetes complications in patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: We analyzed 227 patients with type 1 diabetes using PCR and subsequent cleavage by restriction endonucleases for the promoter variants A-3826G in the UCP1 gene, G-866A in the UCP2 gene, and C-55T in the UCP3 gene. RESULTS: No effect of the A-3826G polymorphism in the UCP1 gene on diabetes complications was found. Patients who were heterozygous or homozygous for the G-866A polymorphism in the UCP2 gene or the C-55T polymorphism in the UCP3 gene had a significantly reduced prevalence of diabetic neuropathy (UCP2: odds ratio 0.44 [95% CI 0.24 0.79], P = 0.007; UCP3: 0.48 [0.25-0.92], P = 0.031), whereas there was no association with other diabetes complications. This effect was stronger when G 866A and C-55T occurred in a cosegregatory manner (UCP2 and UCP3: 0.28 [0.12 0.65], P = 0.002). Furthermore, a multiple logistic regression model showed an age- and diabetes duration-independent effect of the cosegregated polymorphisms on the prevalence of diabetic neuropathy (P = 0.013). CONCLUSIONS: Our data indicate that both the G-866A polymorphism in the UCP2 gene and the C-55T polymorphism in the UCP3 gene are associated with a reduced risk of diabetic neuropathy in type 1 diabetes. Thus, the results presented here support the hypothesis that higher expression of uncoupling protein might prevent mitochondria-mediated neuronal injury and, ultimately, diabetic neuropathy. PMID- 16373903 TI - Effects of different insulin regimes on postprandial myocardial perfusion defects in type 2 diabetic patients. AB - OBJECTIVE: Postprandial glycemia is an independent risk factor for cardiovascular disease that is more powerful than fasting glycemia and determines myocardial perfusion defects in type 2 diabetes. We examined the efficacy of two different insulin regimes (regular insulin and insulin analog) in controlling postprandial hyperglycemia and in preventing myocardial perfusion abnormalities. RESEARCH DESIGN AND METHODS: A total of 20 consecutive type 2 diabetic patients and 20 control subjects were enrolled in this randomized, three-way, cross-over, placebo controlled study. Myocardial perfusion was assessed by myocardial contrast echocardiography (MCE) in fasting and postprandial (120 min) state. RESULTS: Insulin analog was associated with lower, but not statistically significant, postprandial glycemia than regular insulin (glucose increase: 116 +/- 8 vs. 136 +/- 5%, P = NS). However, the area under the curve following insulin analog was significantly lower than regular insulin (18,354 +/- 702 vs. 20,757 +/- 738 mg per 120 min, P = 0.032). Fasting myocardial flow velocity (beta), myocardial blood volume (MBV), and myocardial blood flow (MBF) did not differ between control and type 2 diabetic subjects. Postprandial beta (0.67 +/- 0.24 vs. 0.92 +/- 0.25, P < 0.01), MBV (8.4 +/- 2 vs. 10.9 +/- 1.2, P < 0.01), and MBF (5.6 +/- 2 vs. 9.9 +/- 2.8, P < 0.01) increased significantly in control subjects. In type 2 diabetes, during placebo in the postprandial state, beta increased (0.65 +/- 0.27 vs. 0.89 +/- 0.24, P < 0.01), while MBV (8.34 +/- 1.2 vs. 4.3 +/- 1.3, P < 0.01) and MBF (5.4 +/- 1.5 vs. 3.4 +/- 0.9, P < 0.01) decreased. Similar changes in MCE variables were observed after regular insulin: beta increased (0.65 +/- 0.22 vs. 0.92 +/- 0.12, P < 0.01) and MBV (8.2 +/- 2 vs. 5.2 +/- 1.16, P < 0.01) and MBF (5.4 +/- 1.9 vs. 4.2 +/- 0.86, P < 0.01) were reduced. After insulin analog, postprandial beta (0.66 +/- 0.18 vs. 0.9 +/- 0.18, P < 0.01), MBV (8.2 +/ 1.6 vs. 9.6 +/- 1.2, P < 0.01), and MBF (5.4 +/- 2 vs. 7.2 +/- 1.9, P < 0.01) increased. Values of postprandial MBV and MBF were higher after insulin analog than regular insulin treatment. CONCLUSIONS: Insulin analog partially reversed myocardial perfusion abnormalities observed in postprandial state by improving glucose control. PMID- 16373904 TI - A randomized comparison of pioglitazone to inhibit restenosis after coronary stenting in patients with type 2 diabetes. AB - OBJECTIVE: Recent studies have demonstrated that the treatment with thiazolidinediones reduces in-stent restenosis. The aim of this study was to elucidate the mechanism of the efficacy of pioglitazone for preventing in-stent restenosis in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: We conducted a prospective, randomized trial involving 54 type 2 diabetic patients referred for coronary stenting who were randomly assigned to either the control or the pioglitazone group. Quantitative coronary angiography was performed at study entry and at 6 months follow-up. Endothelial nitric oxide synthase (eNOS), tumor necrosis factor alpha, interleukin-6, leptin, and adiponectin were measured at study entry and at 6 months follow-up. RESULTS: A total of 28 patients were randomly assigned to the control group, and 26 patients were assigned to the pioglitazone group. There were no significant differences in glycemic control levels or in lipid levels in the two groups at baseline or at follow-up. Insulin, homeostasis model assessment of insulin resistance, eNOS, and leptin at follow-up were significantly reduced in the pioglitazone group compared with the control group. The late luminal loss and in-stent restenosis were significantly less in the pioglitazone group than in the control group. Leptin independently correlated with late luminal loss at multiple regression analysis. CONCLUSIONS: The treatment with pioglitazone in type 2 diabetic patients significantly reduced leptin. This decreased leptin improved insulin resistance and endothelial function with the reduction of insulin. The improved endothelial function affected the reduction of in-stent restenosis. PMID- 16373905 TI - Impaired coronary endothelium-dependent vasodilation is associated with microalbuminuria in patients with type 2 diabetes and angiographically normal coronary arteries. AB - OBJECTIVE: Microalbuminuria and impaired endothelium-dependent vasodilation are both predictors for cardiac events in patients with type 2 diabetes. The aim of the study was to evaluate whether microalbuminuria correlated with coronary endothelium-dependent vasodilation. RESEARCH DESIGN AND METHODS: We evaluated 84 patients (47 men, mean age 50.5 +/- 5.9 years) with type 2 diabetes for 9.4 +/- 3.4 years, without angiographic coronary stenosis and without major cardiovascular risk factors or other confounding factors, for endothelium investigation. Quantitative coronary angiography was used to assess coronary artery response to cold pressor testing, used to assess endothelium-dependent vasodilation, and to isosorbide dinitrate (endothelium-independent vasodilation). RESULTS: Endothelium-dependent vasodilation differed in the patients with and without microalbuminuria (changes in coronary artery diameter during cold pressor testing: -15.0 +/- 1.9% vs. -10.2 +/- 1.3%, respectively, P < 0.05) and correlated with urinary albumin excretion rate (r = -0.39, P = 0.003), diastolic blood pressure (r = 0.29, P < 0.01), and left ventricular mass index (r = -0.24, P < 0.05). Independent predictors for endothelium-dependent vasodilation were urinary albumin excretion rate (beta -0.04 [95% CI -0.07 to -0.01], P < 0.005) and left ventricular mass index (-0.26 [-0.49 to -0.05], P < 0.05). Endothelium independent vasodilation was similar in both groups. CONCLUSIONS: Type 2 diabetic patients with microalbuminuria have a more severely impaired coronary endothelium dependent vasodilation than those with normoalbuminuria. These data suggest a common pathophysiological process for both coronary vasomotor abnormalities and microalbuminuria. PMID- 16373906 TI - A single factor underlies the metabolic syndrome: a confirmatory factor analysis. AB - OBJECTIVE: Confirmatory factor analysis (CFA) was used to test the hypothesis that the components of the metabolic syndrome are manifestations of a single common factor. RESEARCH DESIGN AND METHODS: Three different datasets were used to test and validate the model. The Spanish and Mauritian studies included 207 men and 203 women and 1,411 men and 1,650 women, respectively. A third analytical dataset including 847 men was obtained from a previously published CFA of a U.S. population. The one-factor model included the metabolic syndrome core components (central obesity, insulin resistance, blood pressure, and lipid measurements). We also tested an expanded one-factor model that included uric acid and leptin levels. Finally, we used CFA to compare the goodness of fit of one-factor models with the fit of two previously published four-factor models. RESULTS: The simplest one-factor model showed the best goodness-of-fit indexes (comparative fit index 1, root mean-square error of approximation 0.00). Comparisons of one factor with four-factor models in the three datasets favored the one-factor model structure. The selection of variables to represent the different metabolic syndrome components and model specification explained why previous exploratory and confirmatory factor analysis, respectively, failed to identify a single factor for the metabolic syndrome. CONCLUSIONS: These analyses support the current clinical definition of the metabolic syndrome, as well as the existence of a single factor that links all of the core components. PMID- 16373907 TI - Metabolic syndrome: risk factor distribution and 18-year mortality in the multiple risk factor intervention trial. AB - OBJECTIVE: To examine the long-term association of metabolic syndrome with mortality among those at high risk for cardiovascular disease (CVD). RESEARCH DESIGN AND METHODS: A total of 10,950 Multiple Risk Factor Intervention Trial (MRFIT) survivors were followed for mortality an additional median 18.4 years (1980-1999). Proportional hazards models examined multivariate-adjusted risks associated with Adult Treatment Panel III-defined metabolic syndrome conditions, with BMI substituted for waist circumference. RESULTS: At MRFIT annual visit 6, 4,588 (41.9%) men, mean age (+/-SD) 53.0 +/- 5.9 years, had metabolic syndrome and 6,362 did not. Comparing men with metabolic syndrome to men without, adjusted hazard ratios (HRs) were 1.21 (95% CI 1.13-1.29), 1.49 (1.35-1.64), and 1.51 (1.34-1.70) for 18-year total, CVD, and coronary heart disease mortality, respectively. Among men with metabolic syndrome, elevated glucose (1.54 [1.34 1.78]) and low HDL cholesterol (1.45 [1.17-1.54]) were most predictive of CVD mortality, followed by elevated BMI (1.34 [1.17-1.54]), elevated blood pressure (1.25 [0.98-1.58]), and elevated triglycerides (1.06 [0.86-1.30]). In contrast, for men without metabolic syndrome, the HR for low HDL cholesterol was 1.02 (0.86 1.22). Among metabolic syndrome men with no nonfatal CVD event, smokers with elevated LDL cholesterol showed higher CVD mortality (1.79 [1.22-2.63]) compared with nonsmokers without elevated LDL cholesterol; this additional risk was even greater for metabolic syndrome men with a nonfatal CVD event (2.11 [1.32-3.38]). CONCLUSIONS: Metabolic syndrome is associated with an increased risk of mortality. Among those with metabolic syndrome, risk is further increased by having more metabolic syndrome conditions, by cigarette smoking, and by elevated LDL cholesterol. Primary prevention of each metabolic syndrome condition should be emphasized, and presence of each condition should be treated in accordance with current guidelines. PMID- 16373908 TI - Medical mistrust, diabetes self-management, and glycemic control in an indigent population with type 2 diabetes. PMID- 16373909 TI - Age-specific advantages of continuous subcutaneous insulin infusion as compared with multiple daily injections in pediatric patients: one-year follow-up comparison by matched-pair analysis. PMID- 16373910 TI - Effects of exercise and nutrition on memory in Japanese Americans with impaired glucose tolerance. PMID- 16373911 TI - Microencapsulated pancreatic islet allografts into nonimmunosuppressed patients with type 1 diabetes: first two cases. PMID- 16373912 TI - Globular adiponectin activates nuclear factor-kappaB in vascular endothelial cells, which in turn induces expression of proinflammatory and adhesion molecule genes. PMID- 16373913 TI - Increased urinary excretions of immunoglobulin g, ceruloplasmin, and transferrin predict development of microalbuminuria in patients with type 2 diabetes. PMID- 16373915 TI - Metabolic syndrome and asymptomatic peripheral artery disease in subjects over 60 years of age. PMID- 16373914 TI - The new worldwide definition of metabolic syndrome is not a better diagnostic predictor of cardiovascular disease in Japanese diabetic patients than the existing definitions: additional analysis from the Japan Diabetes Complications Study. PMID- 16373916 TI - The lipid accumulation product is better than BMI for identifying diabetes: a population-based comparison. PMID- 16373917 TI - Thiazolidinediones: the case for early use. PMID- 16373918 TI - The Hurricane Katrina aftermath and its impact on diabetes care: observations from "ground zero": lessons in disaster preparedness of people with diabetes. PMID- 16373919 TI - Developments in diabetes and insulin resistance. PMID- 16373921 TI - Retrospective review of metformin in inpatients and outpatients at the University of Michigan. PMID- 16373922 TI - Changes in weight and glucose tolerance during treatment with mirtazapine. PMID- 16373923 TI - The peroxisome proliferator-activated receptor-gamma2 P12A polymorphism and type 2 diabetes in an Arab population. PMID- 16373924 TI - The application of synthetic hANP in diabetic nephropathy with nephrotic syndrome. PMID- 16373925 TI - Seasonality in the incidence of type 2 diabetes: a population-based study. PMID- 16373926 TI - Circulating levels of interleukin-18 independent of body fat and fat-free mass: results from the MONICA/KORA study. PMID- 16373927 TI - Vitamin D deficiency is more common in type 2 than in type 1 diabetes. PMID- 16373929 TI - Introduction. PMID- 16373930 TI - Summary of revisions for the 2006 Clinical Practice Recommendations. PMID- 16373931 TI - Standards of medical care in diabetes--2006. PMID- 16373932 TI - Diagnosis and classification of diabetes mellitus. PMID- 16373933 TI - Diabetes care in the school and day care setting. PMID- 16373934 TI - Diabetes care at diabetes camps. PMID- 16373935 TI - Diabetes management in correctional institutions. PMID- 16373936 TI - Hypoglycemia and employment/licensure. PMID- 16373937 TI - Third-party reimbursement for diabetes care, self-management education, and supplies. PMID- 16373938 TI - Technical reviews. PMID- 16373939 TI - National standards for diabetes self-management education. PMID- 16373940 TI - The glycemic and insulinemic index of plain sweet biscuits: relationships to in vitro starch digestibility. AB - BACKGROUND: Among the various classes of processed starchy foods, wheat-based cereal products exhibit a wide range in glycemic and insulinemic responses. Understanding starch behavior during cooking and processing may help identify strategies that lower postprandial glycemia and insulinemia. OBJECTIVE: To determine the relationship between the in vivo glycemic index (GI)/insulinemic index (II) and in vitro digestibility and composition characteristics (rapidly available glucose and slowly available glucose, RAG and SAG respectively) of 24 plain sweet biscuits (cookies). METHODS: The products were commercially available and selected on the basis of their high starch content. In vivo responses (GI and II) were measured by standardised methods over 7 studies, with 12 subjects in each study (30 males, 42 females). In vitro digestibility characteristics were measured by the Englyst procedure. RESULTS: The observed GI ranged from 38 to 60 (low to moderate) with the majority between 40 and 50, and correlated strongly with the observed insulinemic index (r = 0.76, P < 0.0001). The digestibility profile of carbohydrates was significantly correlated to in vivo responses (SAG and GI: r = -0.41; p = 0.04; SAG and II: r = -0.52; p < 0.01; RAG and GI: r = 0.5; p = 0.01; RAG and II: r = 0.34; p = 0.1) and explained in vivo responses better than fat, protein and fiber content amongst this selection of plain sweet biscuits. CONCLUSION: The findings indicate that plain sweet biscuits have a low GI and a moderate II and that these characteristics are correlated to in vitro starch digestibility and are dependent on the type of processing. PMID- 16373941 TI - Airway and circulating levels of carotenoids in asthma and healthy controls. AB - BACKGROUND: Elevated oxidative stress and impaired antioxidant defences are increasingly recognised features of asthma. Carotenoids are potent dietary antioxidants that may protect against asthma by reducing oxidative damage. OBJECTIVES: This study aimed firstly, to characterise circulating and airway levels of carotenoids in asthma compared to healthy controls, in relation to dietary intake. Secondly, the study aimed to test whether airway lycopene defences can be improved using oral supplements. METHODS: Induced sputum and peripheral blood samples were collected from subjects with asthma (n = 15) and healthy controls (n = 16). Dietary carotenoid intakes were estimated using the 24 hour recall method and analysed using a modified version of the Foodworks 210 Nutrient Calculation Software. Another group of healthy controls (n = 9) were supplemented with 20 mg/day lycopene for 4 weeks. Carotenoids (beta-carotene, lycopene, alpha-carotene, beta-cryptoxanthin, lutein/zeaxanthin) were measured by HPLC. RESULTS: Despite similar dietary intake, whole blood levels of total carotenoids, lycopene, lutein, beta-cryptoxanthin, alpha-carotene and beta carotene were significantly lower in asthma than controls. However, there were no differences in plasma or sputum carotenoid levels. Induced sputum carotenoid levels were significantly lower than plasma and whole blood levels, but correlated strongly with plasma levels (r = 0.798, p < 0.001). Although there were no overall increases in either plasma or sputum lycopene levels following supplementation, changes in airway lycopene levels correlated with changes in plasma levels (r = 0.908, p < 0.002). CONCLUSIONS: Whole blood, but not plasma or sputum, carotenoid levels are deficient in asthma. Plasma carotenoid levels reflect airway carotenoid levels and when plasma levels are improved using oral supplements this is reflected in the airways. PMID- 16373942 TI - Factors associated with obesity in an adult Mediterranean population: influence on plasma lipid profile. AB - OBJECTIVE: The aim of the study was to identify factors associated with obesity, and their influence on plasma lipid profile in an adult Mediterranean population. DESIGN: The data were obtained from a cross-sectional epidemiological survey. SETTING: The study population resided in Andalusia, a western Mediterranean region in southern Spain. SUBJECTS: The survey was carried out with a random sample of 3421 subjects (1747 men, 1674 women) between 25 and 60 years of age. Blood samples were obtained for biochemical assays in a random subsample of 340 subjects (167 men, 173 women). INTERVENTIONS: Food consumption was assessed by 48 h recall. Height, weight, triceps, biceps, subscapular and suprailiac skinfolds, mid-upper arm, waist (WC) and hip circumferences, glucose, total cholesterol, HDL cholesterol, LDL-cholesterol and triglycerides were measured. Information about lifestyles was obtained with a questionnaire. RESULTS: Of the adult population we studied, 18.9% were obese (body mass index [BMI] > or = 30 kg/m2). A larger proportion of men than women were overweight, but the opposite was found for obesity. Mean plasma lipid values were not modified significantly by obesity or lifestyle factors, and were within the normal range. Sex, age, physical exercise and lower educational level were associated directly with the risk of obesity, and smoking was associated inversely with the risk of obesity. In obese smokers WC and waist-hip ratio were larger, and levels of HDL-cholesterol were lower (p < 0.05) than in obese nonsmokers. Glucemia was higher in obese persons who consumed alcohol (p < 0.05) than in obese persons who did not consume alcohol. The risk of hypercholesterolemia and high levels of LDL-cholesterol was associated only with age, and the risk of low levels of HDL-cholesterol was associated only with high WC. CONCLUSION: Our results provide an estimate of the prevalence of obesity in the adult population in southern Spain, and of the associated factors. Sex, age, leisure-time physical exercise and educational level appear to influence obesity. Only age and WC but not BMI were associated with a risk of dyslipidemia. No dietary associations were observed between energy or macronutrient intake and plasma lipid concentrations in overweight or obese persons. PMID- 16373943 TI - Reproducibility and validity of a food frequency questionnaire among Vietnamese in Ho Chi Minh City. AB - OBJECTIVE: Our objective was to assess the reproducibility and validity of a food frequency questionnaire (FFQ) among individuals living in Ho Chi Minh City, Vietnam. METHODS: A total of 116 food items in our FFQ were selected by ranking food items according to their contribution to the population intake of nutrients, based on multiple 24-hour recalls (24HRs) from 144 participants in July 2001. The FFQ was validated by using three 24HRs for 118 men and women in Ho Chi Minh City, Vietnam in August 2002. The reproducibility of the FFQ was assessed by repeated administration at 7 +/- 3.4 days. RESULTS: The reproducibility study showed that Spearman correlation coefficients, adjusted for energy intake, ranged from 0.47 to 0.72, and that weighted kappa values ranged from 0.42 for lipid to 0.65 for potassium. In the validation study, correlation coefficients, adjusted for energy intake, between the FFQ and the 24HRs ranged from 0.16 (calcium) to 0.45 (retinol). The proportion of subjects classified by the FFQ into the same third as determined by the 24HRs was between 42% and 62%. CONCLUSIONS: This food frequency questionnaire measured the usual intake of major nutrients for Vietnamese adults living in Ho Chi Minh City and its urban area, where dietary patterns are similar to those of the Vietnamese population. PMID- 16373944 TI - Time course of oxidative stress status in the postprandial and postabsorptive states in type 1 diabetes mellitus: relationship to glucose and lipid changes. AB - OBJECTIVE: The aim of this study was to compare oxidative stress status (OSS) with blood glucose and lipid changes during the fasting, postprandial and postabsorptive phases in type 1 diabetes mellitus. METHODS: Twenty-three patients on intensive insulin treatment received a standard fat-rich breakfast and lunch. OSS was monitored at fasting (F), just after the post-breakfast glycemia peak (BP) (identified by continuous subcutaneous glucose monitoring), 3.5-h post breakfast (B3.5), just after the post-lunch peak (LP), just after the post-lunch dale (LD) and 5 hours after lunch (L5). RESULTS: Whereas whole blood glutathione and plasma protein thiols increased in the postprandial period (from 6.52 +/- 1.20 (F) to 7.08 +/- 1.45 micromol/g Hb (BP), p = 0.005), ascorbate decreased gradually from 44 +/- 17 (F) to 39 +/- 19 micromol/L (LD), p = 0.015. Retinol and alpha-tocopherol also decreased from 27.1 +/- 7.0 (F) to 25.3 +/- 5.2 micromol/L (BP), p = 0.005. Uric acid decreased later, from 213 +/- 77 (BP) to 204 +/- 68 micromol/L (B3.5), p = 0.01, but then increased in LP (231 +/- 70 micromol/L) and LD to values higher than F (215 +/- 64, micromol/L, p = 0.01). Malondialdehyde increased gradually from 1.02 +/- 0.36 (F) to a maximum of 1.14 +/- 0.40 micromol/L (LP). In the postabsorptive phase (L5) all parameters except for thiols reverted to fasting concentrations. CONCLUSIONS: In type 1 diabetes lipid peroxidation increases during the postprandial phase in parallel to glucose and triglyceride changes. Blood antioxidants, however, followed diverse patterns of change. PMID- 16373945 TI - Weight loss favorably modifies anthropometrics and reverses the metabolic syndrome in premenopausal women. AB - OBJECTIVE: To determine the effects of a weight loss program, including dietary modifications, increased physical activity and dietary supplement (L-carnitine or placebo) on anthropometrics, leptin, insulin, the metabolic syndrome (MS) and insulin resistance in overweight /obese premenopausal women. METHODS: Participants consumed a hypocaloric diet; 30% protein, 30% fat and 40% carbohydrate in addition to increasing number of steps/day. Carnitine supplementation followed a randomized double blind protocol. Protocol lasted for 10 weeks. Seventy subjects (35 in the control and 35 in the carnitine group) completed the intervention. Anthropometrics, plasma insulin and leptin concentrations and body composition were measured. The number of subjects with the MetSyn and insulin resistance, were assessed at baseline and post intervention. RESULTS: Because there were no significant differences between the carnitine and the placebo groups for all measured parameters, participants were grouped together for all analysis. Subjects decreased total energy (-26.6%, p < 0.01) and energy from carbohydrate (-17.3%, p < 0.01) and increased energy from protein by 67% (p < 0.01) and number of steps/day (42.6%, p < 0.01). Body weight (-4.6%, p < 0.001), body mass index (-4.5%, p < 0.01), waist circumference ( 6.5%, p < 0.01), total fat mass (-1.7%, p < 0.01), trunk fat mass (-2.0%, p < 0.01), insulin (- 17.9%, p < 0.01) and leptin (-5.9%, p < 0.05) decreased after the intervention. Ten of 19 participants with insulin resistance became insulin sensitive and 7 of 8 participants with the MetSyn no longer had the syndrome after the intervention. CONCLUSION: Moderate increases in physical activity and a hypocaloric/high protein diet resulted in multiple beneficial effects on body anthropometrics and insulin sensitivity. Realistic dietary and physical activity goals must be the focus of intervention strategies for overweight and obese individuals. PMID- 16373946 TI - Chronic zinc deficiency in mice disrupted T cell lymphopoiesis and erythropoiesis while B cell lymphopoiesis and myelopoiesis were maintained. AB - OBJECTIVE: The purpose of this study was to determine the impact of chronic zinc deficiency (ChrZD) on T and B cell lymphopoiesis, myelopoiesis and erythropoiesis in mice. METHODS: Young adult mice were fed a zinc adequate (ZA) or ChrZD synthetic diet for 34, 45, and 50 days. The cellular composition of the thymus and marrow were determined to assess the impact of ChrZD on lymphopoietic and hematopoietic processes using flow cytometry. Body weights, serum zinc and corticosterone (Cs) were monitored. RESULTS: For ChrZD mice growth was reduced 10% and serum zinc declined 15% by d 34 compared to ZA mice. By d 50 a 25% decrease in growth and 70% depression in serum zinc was noted though there was never any significant reduction in diet intake. Corticosterone rose 2.5 fold by d 34 and remained elevated in ChrZD mice indicating induction of the stress axis. At d 34 the thymus of ChrZD mice was normal but by d 50 there was a 50% cell loss and a 10% reduction in the proportion of Pre-T cells. Most importantly there was a 60% increase in Pre-T cells undergoing apoptosis in ChrZD mice. Pro-T, T helper, and T cytolytic populations were more resistant to ChrZD. Bone marrow cellularity and granulocyte, monocyte, and lymphocyte compartments remained unchanged in ChrZD mice. However, the erythroid compartment was reduced by 35% at d 50. CONCLUSIONS: The thymus was the most sensitive primary tissue to ChrZD. By d 50 it had atrophied by 36% with significant loss of Pre-T cells via apoptosis such that T-cell lymphopoiesis was disrupted. Significant reductions were also noted in the erythropoietic population by d 50. Conversely the marrow maintained myelopoiesis and B cell lymphopoiesis for the 50 d period indicating greater ability to survive a chronic zinc deficiency and exposure to Cs. The anemia and T cell lymphopenia associated with ChrZD in both rodents and humans may result from a greater sensitivity of their precursor cells to zinc deficiency and elevated Cs. PMID- 16373947 TI - Serum homocysteine concentration of US adults associated with fortified cereal consumption. AB - BACKGROUND: Elevated serum total homocysteine (tHcy) concentration is implicated in the etiology of cardiovascular disease (CVD). A significant food source of B vitamins involved in homocysteine metabolism is ready-to-eat cereal (RTEC) in the U.S. OBJECTIVE: To test the hypothesis that tHcy concentration is inversely associated with RTEC intake and blood B-vitamin levels in the U.S. general population. DESIGN: A cross-sectional study using data from the National Health and Nutrition Examination Survey (NHANES 1999-2000). Data were stratified according to age and gender. Men and women > or = 19 y (n = 4,218) were classified as RTEC consumers (RTEC-C; n = 824) and RTEC non-consumers (RTEC-NC; n = 3,394) based on 24-hr dietary recall. RESULTS: Forty nine percent of participants showed folate intake with below the estimated average requirements (EARS). Serum folate and red blood cell (RBC) folate concentrations were increased with age in both genders, and significantly higher among RTEC-C than RTEC-NC (p < 0.05). Mean tHcy concentration increased with age, and was significantly lower among both men and women RTEC-C than among RTEC-NC. In multivariate linear regression analyses, RTEC consumption strongly predicted serum folate and tHcy concentrations. CONCLUSION: tHcy concentrations were significantly lower in RTEC-C among the majority of age/gender groups than in RTEC-NC. RTEC consumption may potentially reduce the risk for CVD, mediated through tHcy. PMID- 16373948 TI - Short-term effect of eggs on satiety in overweight and obese subjects. AB - OBJECTIVE: To test the hypotheses that among overweight and obese participants, a breakfast consisting of eggs, in comparison to an isocaloric equal-weight bagel based breakfast, would induce greater satiety, reduce perceived cravings, and reduce subsequent short-term energy intake. SUBJECTS: Thirty women with BMI's of at least 25 kg/M2 between the ages of 25 to 60 y were recruited to participate in a randomized crossover design study in an outpatient clinic setting. DESIGN: Following an overnight fast, subjects consumed either an egg or bagel-based breakfast followed by lunch 3.5 h later, in random order two weeks apart. Food intake was weighed at breakfast and lunch and recorded via dietary recall up to 36 h post breakfast. Satiety was assessed using the Fullness Questionnaire and the State-Trait Food Cravings Questionnaire, state version. RESULTS: During the pre-lunch period, participants had greater feelings of satiety after the egg breakfast, and consumed significantly less energy (kJ; 2405.6 +/- 550.0 vs 3091.3 +/- 445.5, Egg vs Bagel breakfasts, p < 0.0001), grams of protein (16.8 +/- 4.2 vs 22.3 +/- 3.4, Egg vs Bagel breakfasts, p < 0.0001), carbohydrate 83.1 +/- 20.2 vs 110.9 +/- 18.7, Egg vs Bagel breakfasts, p < 0.0001), and fat 19.4 +/- 5.1 vs 22.8 +/- 3.2, Egg vs Bagel breakfasts, p < 0.0001) for lunch. Energy intake following the egg breakfast remained lower for the entire day (p < 0.05) as well as for the next 36 hours (p < 0.001). CONCLUSIONS: Compared to an isocaloric, equal weight bagel-based breakfast, the egg-breakfast induced greater satiety and significantly reduced short-term food intake. The potential role of a routine egg breakfast in producing a sustained caloric deficit and consequent weight loss, should be determined. PMID- 16373949 TI - Selenomethionine prevents degeneration induced by overexpression of wild-type human alpha-synuclein during differentiation of neuroblastoma cells. AB - OBJECTIVE: High levels of wild-type alpha-synuclein are found in autopsied brain samples of idiopathic Parkinson's disease (PD), some familial PD, some Alzheimer's disease (AD) and Down's syndrome with dementia. Therefore, we have investigated whether overexpression of wild-type alpha-synuclein causes degeneration during adenosine, 3',5'-cyclic monophosphate (cAMP)-induced differentiation of murine neuroblastoma (NB) cells in culture. We have also studied whether selenomethionine can modify the effect of overexpression of alpha synuclein during differentiation of NB cells. METHODS: To study these issues, we established a murine neuroblastoma (NB) clone (NBP2-PN54-C20) that expressed high levels of wild-type human alpha-synuclein as determined by real time PCR and Western blot. We have utilized RO20-1724, an inhibitor of cyclic nucleotide phosphodiesterase, and prostaglandin A1 (PGA1), a stimulator of adenylate cyclase, or RO20-1724 and dibutyryl cAMP to induce terminal differentiation in over 95% of the cell population by elevating the intracellular levels of cAMP in NB cells. The viability of cells was determined by MTT assay and LDH leakage assay, and the degeneration was documented by photomicrographs. RESULTS: The results showed that overexpression of human wild-type alpha-synuclein decreased viability and increased degenerative changes in comparison to those observed in vector control cells, when differentiation was induced by treatment with RO20 1724 and PGA1, but not with RO20-1724 and dibutyryl cAMP. When selenomethionine was added to NB cells overexpressing alpha-synuclein immediately after the addition of RO20-1724 and PGA1, the viability and degenerative changes were markedly reduced, suggesting the involvement of increased oxidative stress in the mechanism of action of alpha-synuclein. This protective effect was not observed after treatment with sodium selenite or methionine. CONCLUSIONS: Data suggested that Overexpression of wild-type human alpha-synuclein-decreased viability and increased the levels of degenerative changes during differentiation of NB cells were reduced by selenomethionine treatment. This suggest that one of mechanisms of action alpha-synuclein may involve increased oxidative stress. PMID- 16373952 TI - Dietary protein: an essential nutrient for bone health. AB - Nutrition plays a major role in the development and maintenance of bone structures resistant to usual mechanical loadings. In addition to calcium in the presence of an adequate vitamin D supply, proteins represent a key nutrient for bone health, and thereby in the prevention of osteoporosis. In sharp opposition to experimental and clinical evidence, it has been alleged that proteins, particularly those from animal sources, might be deleterious for bone health by inducing chronic metabolic acidosis which in turn would be responsible for increased calciuria and accelerated mineral dissolution. This claim is based on an hypothesis that artificially assembles various notions, including in vitro observations on the physical-chemical property of apatite crystal, short term human studies on the calciuric response to increased protein intakes, as well as retrospective inter-ethnic comparisons on the prevalence of hip fractures. The main purpose of this review is to analyze the evidence that refutes a relation of causality between the elements of this putative patho-physiological "cascade" that purports that animal proteins are causally associated with an increased incidence of osteoporotic fractures. In contrast, many experimental and clinical published data concur to indicate that low protein intake negatively affects bone health. Thus, selective deficiency in dietary proteins causes marked deterioration in bone mass, micro architecture and strength, the hallmark of osteoporosis. In the elderly, low protein intakes are often observed in patients with hip fracture. In these patients intervention study after orthopedic management demonstrates that protein supplementation as given in the form of casein, attenuates post-fracture bone loss, increases muscles strength, reduces medical complications and hospital stay. In agreement with both experimental and clinical intervention studies, large prospective epidemiologic observations indicate that relatively high protein intakes, including those from animal sources are associated with increased bone mineral mass and reduced incidence of osteoporotic fractures. As to the increased calciuria that can be observed in response to an augmentation in either animal or vegetal proteins it can be explained by a stimulation of the intestinal calcium absorption. Dietary proteins also enhance IGF-1, a factor that exerts positive activity on skeletal development and bone formation. Consequently, dietary proteins are as essential as calcium and vitamin D for bone health and osteoporosis prevention. Furthermore, there is no consistent evidence for superiority of vegetal over animal proteins on calcium metabolism, bone loss prevention and risk reduction of fragility fractures. PMID- 16373953 TI - The role of dairy foods in weight management. AB - Dietary calcium appears to play a pivotal role in the regulation of energy metabolism and obesity risk. High calcium diets attenuate body fat accumulation and weight gain during periods of over-consumption of an energy-dense diet and to increase fat breakdown and preserve metabolism during caloric restriction, thereby markedly accelerating weight and fat loss. This effect is mediated primarily by circulating calcitriol, which regulates adipocyte intracellular Ca(2+). Studies of human adipocyte metabolism demonstrate a key role for intracellular Ca(2+) in regulating lipid metabolism and triglyceride storage, with increased intracellular Ca(2+) resulting in stimulation of lipogenic gene expression and lipogenesis and suppression of lipolysis, resulting in adipocyte lipid filling and increased adiposity. Moreover, the increased calcitriol produced in response to low calcium diets stimulates adipocyte Ca(2+) influx and, consequently, promotes adiposity, while higher calcium diets inhibit lipogenesis, promote lipolysis, lipid oxidation and thermogenesis and inhibit diet-induced obesity in mice. Notably, dairy sources of calcium exert markedly greater effects in attenuating weight and fat gain and accelerating fat loss. This augmented effect of dairy products versus supplemental calcium has been localized, in part, to the whey fraction of dairy and is likely due to additional bioactive compounds, such as angiotensin converting enzyme (ACE) inhibitors in dairy, as well as the rich concentration of branched chain amino acids, which act synergistically with calcium to attenuate adiposity; however, these compounds do not fully account for the observed effects, as whey has significantly greater bioactivity than found in these compounds. These concepts are confirmed by epidemiological data as well as recent clinical trials which demonstrate that diets which include at least three daily servings of dairy products result in significant reductions in body fat mass in obese humans in the absence of caloric restriction and markedly accelerates the weight and body fat loss secondary to caloric restriction compared to low dairy diets. These data indicate an important role for dairy products in both the ability to maintain a healthy weight and the management of overweight and obesity. PMID- 16373954 TI - Milk consumption does not lead to mucus production or occurrence of asthma. AB - There is a belief among some members of the public that the consumption of milk and dairy products increases the production of mucus in the respiratory system. Therefore, some who believe in this effect renounce drinking milk. According to Australian studies, subjects perceived some parameters of mucus production to change after consumption of milk and soy-based beverages, but these effects were not specific to cows' milk because the soy-based milk drink with similar sensory characteristics produced the same changes. In individuals inoculated with the common cold virus, milk intake was not associated with increased nasal secretions, symptoms of cough, nose symptoms or congestion. Nevertheless, individuals who believe in the mucus and milk theory report more respiratory symptoms after drinking milk. In some types of alternative medicine, people with bronchial asthma, a chronic inflammatory disease of the lower respiratory tract, are advised not to eat so-called mucus-forming foods, especially all kinds of dairy products. According to different investigations the consumption of milk does not seem to exacerbate the symptoms of asthma and a relationship between milk consumption and the occurrence of asthma cannot be established. However, there are a few cases documented in which people with a cow's milk allergy presented with asthma-like symptoms. PMID- 16373955 TI - Dairy product consumption and the risk of breast cancer. AB - It has been suggested in some reports that dairy product consumption may increase the risk of breast cancer. This review gives a brief overview of the etiology of breast cancer and in particular the roles of fat, bovine growth hormone, insulin like growth factor-1 and estrogens. Evidence from animal studies and epidemiology does not support a role for fat in the etiology of breast cancer. The daily intake of insulin-like growth factor-1 and biologically active estrogens from dairy products is minute in comparison to the daily endogenous secretion of these factors in women, whereas bovine growth hormone is biologically inactive in humans. On the other hand, milk contains rumenic acid, vaccenic acid, branched chain fatty acids, butyric acid, cysteine-rich whey proteins, calcium and vitamin D; components, which have the potential to help prevent breast cancer. Evidence from more than 40 case-control studies and 12 cohort studies does not support an association between dairy product consumption and the risk of breast cancer. PMID- 16373956 TI - The myth of increased lactose intolerance in African-Americans. AB - In the United States, approximately three fourths of African-Americans have the potential for symptoms of lactose intolerance because lactose digestion depends on the presence of the enzyme lactase-phlorizin hydrolase which is reduced by up to 90-95% in individuals with lactase nonpersistence. The 'African-American diet' is more likely to be low in a variety of vitamins and minerals, including calcium. African-Americans consume low amounts of dairy foods and do not meet recommended intakes of a variety of vitamins and minerals, including calcium. Low intake of calcium and other nutrients put African-Americans at an increased risk for chronic diseases. The 2005 Dietary Guidelines recommend consuming three servings of dairy foods per day to ensure adequate calcium intake, among other nutrients, and the National Medical Association has recently published a similar recommendation of three to four servings of dairy per day for the African American population. Research has shown that lactose maldigesters, including African-American maldigesters, can consume at least one cup (8 oz) of milk without experiencing symptoms, and that tolerance can be improved by consuming the milk with a meal, choosing yogurt or hard cheeses, or using products that aid in the digestion of lactose such as lactase supplements or lactose-reduced milks. PMID- 16373957 TI - Newer perspectives on calcium nutrition and bone quality. AB - It is now generally accepted that an adequate calcium intake is important for building and maintaining a skeleton that expresses quantitatively the full genetic program and reduces lifetime fracture risk. In this brief review we focus mainly on a new and growing body of evidence indicating a benefit of adequate calcium intake on qualitative features of the skeleton that, independent of the quantity of bone, themselves influence skeletal strength and fragility. Change in bone mass and size during growth are dependent on both calcium intake and exercise, with the largest differences being observed in prepubertal children who have both high exercise levels and high calcium intakes. Much of this benefit is expressed as increased bone diameter (and hence stiffness). Fracture risk peaks at about the time of puberty and is inversely related to bone mass. However, even prepubertally, children with low calcium intakes have been reported to have a fracture rate 2.7x that of their birth cohort. Bone remodeling triples from age 50 to 65 in typical women and is now recognized to have primarily a homeostatic basis. While remodeling improves bone strength by repairing acquired defects, homeostatic remodeling, while necessary to maintain blood calcium levels, contributes only structural weakness to bone. High calcium intakes in postmenopausal and older women reduce this homeostatic remodeling to approximately pre-menopausal values and improve bone strength immediately, well prior to any appreciable change in bone mass. PMID- 16373958 TI - Cow's milk allergy: a complex disorder. AB - Cow's milk allergy (CMA) is a complex disorder. Numerous milk proteins have been implicated in allergic responses and most of these have been shown to contain multiple allergenic epitopes. There is considerable heterogeneity amongst allergic individuals for the particular proteins and epitopes to which they react, and to further complicate matters, allergic reactions to cow's milk are driven by more than one immunological mechanism. Finally, the incidence and dominant allergic mechanisms change with age, with IgE-mediated reactions common in infancy and non-IgE-mediated reactions dominating in adults. The complexity of CMA has lead to many public misconceptions about this disorder, including confusion with lactose intolerance and frequent self-misdiagnosis. Indeed, the prevalence of self-diagnosed CMA in the community is 10-fold higher than the clinically proven incidence, suggesting a sizable population is unnecessarily eschewing dairy products. Avoidance of dairy foods, whether for true or perceived CMA, carries with it nutritional consequences and the provision of appropriate nutritional advice is important. In this review, the epidemiology and natural course of CMA is discussed along with our current understanding of its triggers and immunological mechanisms. We examine current strategies for the primary and secondary prevention of allergic sensitization and the ongoing search for effective therapies to ultimately cure CMA. PMID- 16373959 TI - Pharmaceutical expenditure compared across countries. AB - Pharmaceutical expenditures for 20 member countries of the Organisation for Economic Co-Operation and Development (OECD) are discussed and the appropriateness of making comparisons amongst these countries is addressed. This is a summary of a lecture presented at the International Conference on Pharmacoepidemiology in 2004. PMID- 16373960 TI - Chromosomes and cancer, Boveri revisited. PMID- 16373961 TI - Predictive testing of early CIN behaviour by molecular biomarkers. AB - Each year, 330,000 new Cervical Intraepithelial Neoplasias(CIN) occur in the European Union (EU) of which 120,000 are early CIN where grade (1, 2) indicates the progression-risk to CIN-3 and therefore determines the treatment choice. However, the Positive Predictive Value (PPV) of CIN grade to predict progression is low (10% and 20% for CIN-1 and -2 respectively, 16% on average) resulting in an enormous number of over-treatments indicating worrisome grade reproducibility.Certain molecular biomarkers such as Ki-67 have a higher PPV (30%, an improvement of 14%), which in Europe alone could improve treatment for many thousands of women per year with considerable cost reduction for the health care system. The quantitative Ki-67 prognostic model has been validated in independent retrospective and prospective studies from different laboratories. Moreover, the PPV of Ki-67 alone can be improved by additional molecular biomarkers (retinoblastoma protein = Rb, cytokeratins= CK-14/-13). Combined Ki67 Rb allows a 2-tiered progression-risk subgroup assignment as very low ( approximately 0% progression, 71% of all CIN-I/II patients)and high risk (48% progression risk, incidence 32%), leaving a small (7% of all) prognostically undetermined group (17% progression). Additional CK-14 and -13 analysis can sub classify the high-risk in an intermediate and very high risk subgroup(with 40% and 100% progression risks respectively).Thus, molecular biomarkers are potentially important determinators of early CIN lesion behaviour. Important factors for widespread acceptance of molecular biomarkers are (1) market penetration by user-friendly equipment, (2) (inter)national keeping of GLP conditions (reproducibility, independent validation), requiring customer-driven industrial efforts,governmental measures, and additional PPV improvement to further reduce over-treatment. PMID- 16373962 TI - Genetic predisposition to sporadic cancer: how to handle major effects of minor genes? AB - Predisposition to non-familial, sporadic cancer is strongly influenced by multiple tumor susceptibility genes (TSGs), each with apparently minor effects on the cancer phenotype. Sequence analysis of the human genome has yielded numerous single nucleotide polymorphisms (SNPs), raising the expectation that new low penetrance TSGs will be identified that can be used to estimate an individuals cancer risk. However, mouse models for human cancer showed that the effects of many low-penetrance TSGs are highly variable due to their involvement in epistatic interactions. Together, these interacting TSGs form large molecular networks, which represent cancer-associated biological modules that influence the tumorigenic process. As a consequence, although allelic variation in one TSG on a permissive genetic background can have major effects on tumor development, the net effect of allelic variation in multiple interacting TSGs remains hard to predict. Therefore, the predictive value of SNP-analysis to estimate an individuals cancer risk will be restricted to those TSGs that exhibit single-gene effects. New strategies need to be developed to evaluate cancer risk associated with biological modules that are influenced by TSG-networks. PMID- 16373963 TI - The chromosomal basis of cancer. AB - Conventional genetic theories have failed to explain why cancer (1) is not heritable and thus extremely rare in newborns, (2) is caused by non-mutagenic carcinogens, (3) develops only years to decades after initiation by carcinogens, (4) follows pre-neoplastic aneuploidy, (5) is aneuploid, (6) is chromosomally and phenotypically "unstable", (7) carries specific aneusomies, (8) generates much more complex phenotypes than conventional mutation such as multidrug resistance, (9) generates nonselective phenotypes such as metastasis (no benefit at native site) and "immortality" (not necessary for tumorigenesis), and (10) does not contain carcinogenic mutations. We propose, instead, that cancer is a chromosomal disease. Accordingly carcinogenesis is initiated by random aneuploidies, which are induced by carcinogens or spontaneously. Since aneuploidy unbalances 1000s of genes, it corrupts teams of proteins that segregate, synthesize and repair chromosomes. Aneuploidy is therefore a steady source of chromosomal variations from which, in classical Darwinian terms, selection encourages the evolution and malignant progression of cancer cells. The rates of specific chromosomal variations can exceed conventional mutations by 4-11 orders of magnitude, depending on the degrees of aneuploidy. Based on their chromosomal constitution cancer cells are new cell "species" with specific aneusomies, but unstable karyotypes. The cancer-specific aneusomies generate complex, malignant phenotypes through the abnormal dosages of 1000s of genes, just as trisomy 21 generates Down syndrome. In sum, cancer is caused by chromosomal disorganization, which increases karyotypic entropy. Thus, cancer is a chromosomal rather than a genetic disease. The chromosomal theory explains (1) non-heritable cancer because aneuploidy is not heritable, (2) non-mutagenic carcinogens as aneuploidogens, (3) long neoplastic latencies by the low probability of evolving new species, (4) nonselective phenotypes via genes hitchhiking with selective chromosomes, and (5) immortality because, through their cellular heterogeneity, cancers survive negative mutations and cytotoxic drugs via resistant subspecies. PMID- 16373964 TI - Molecular characterization of PDGFR-alpha/PDGF-A and c-KIT/SCF in gliosarcomas. AB - Gliosarcomas are rare and poorly characterized malignant brain tumors that exhibit a biphasic tissue pattern with areas of gliomatous and sarcomatous differentiation. These tumors are histological variants of glioblastoma, displaying a similar genetic profile and dismal prognosis. Up-regulation of PDGFR subfamily of tyrosine kinase members, PDGFR-alpha and c-Kit, and their intracellular effectors RAS/RAF/MAPK has a crucial role in the cancer development. In addition, signal transduction mediated by activating mutations of c-Kit and PDGFR can be effectively blocked by specific tyrosine kinase inhibitors, such as Imatinib mesylate. The aim of this study was to characterize the molecular alterations of PDGFR signaling in gliosarcomas. Six cases were analyzed by immunohistochemistry for the expression of PDGFR-alpha, c-Kit and their ligands PDGF-A and SCF, respectively. The cases were further evaluated for the presence of activating mutations of PDGFR-alpha (exons 12 and 18) and c-kit (exons 9, 11, 13, and 17), as well as B-RAF (exons 11 and 15). Expression of PDGF A was found in all cases and co-expression of PDGFR-alpha was observed in three cases. Four cases showed expression of SCF, and c-Kit was observed only in one case that also expressed SCF. Generally, immunoreaction predominates in the glial component. The mutational analysis of PDGFR-alpha showed the presence of an IVS17 50insT intronic insertion in two cases, one of them also with a 2472C > T silent mutation; this silent mutation was also found in another case. Glioma cell line analysis of IVS17-50insT insertion showed no influence on PDGFR-alpha gene splicing. No mutations were detected in c-kit and B-RAF oncogenes. Our results indicate that activating mutations of PDGFR-alpha, c-kit and B-RAF are absent in gliosarcomas. Nevertheless, the presence of a PDGFR-a/PDGFA and c-Kit/SCF autocrine/paracrine stimulation loop in a proportion of cases, supports the potential role of specific tyrosine kinase inhibitors in the treatment of gliosarcomas. PMID- 16373965 TI - Biphasic chromatin structure and FISH signals in reflect intranuclear order. AB - BACKGROUND AND AIM: One of the two parental allelic genes may selectively be expressed, regulated by imprinting, X-inactivation or by other less known mechanisms. This study aims to reflect on such genetic mechanisms. MATERIALS AND METHODS: Slides from short term cultures or direct smears of blood, bone marrow and amniotic fluids were hybridized with FISH probes singly, combined or sequentially. Two to three hundred cells were examined from each preparation. RESULTS AND SIGNIFICANCE: A small number of cells (up to about 5%), more frequent in leukemia cases, showed the twin features: (1) nuclei with biphasic chromatin, one part decondensed and the other condensed; and (2) homologous FISH signals distributed equitably in those two regions. The biphasic chromatin structure with equitable distribution of the homologous FISH signals may correspond to the two sets of chromosomes, supporting observations on ploidywise intranuclear order. The decondensed chromatin may relate to enhanced transcriptions or advanced replications. CONCLUSIONS: Transcriptions of only one of the two parental genomes cause allelic exclusion. Genomes may switch with alternating monoallelic expression of biallelic genes as an efficient genetic mechanism. If genomes fail to switch, allelic exclusion may lead to malignancy. Similarly, a genome-wide monoallelic replication may tilt the balance of heterozygosity resulting in aneusomy, initiating early events in malignant transformation and in predicting cancer mortality. PMID- 16373966 TI - Development of 3D chromatin texture analysis using confocal laser scanning microscopy. AB - INTRODUCTION: Analysis of nuclear texture features as a measure of nuclear chromatin changes has been proven to be useful when measured on thin (5-6 microm) tissue sections using conventional 2D bright field microscopy. The drawback of this approach is that most nuclei are not intact because of those thin sections. Confocal laser scanning microscopy (CLSM) allows measurements of texture in 3D reconstructed nuclei. The aim of this study was to develop 3D texture features that quantitatively describe changes in chromatin architecture associated with malignancy using CLSM images. METHODS: Thirty-five features thoughtfully chosen from 4 categories of 3D texture features (discrete texture features, Markovian features, fractal features, grey value distribution features) were selected and tested for invariance properties (rotation and scaling) using artificial images with a known grey value distribution. The discriminative power of the 3D texture features was tested on artificially constructed benign and malignant 3D nuclei with increasing nucleolar size and advancing chromatin margination towards the periphery of the nucleus. As a clinical proof of principle, the discriminative power of the texture features was assessed on 10 benign and 10 malignant human prostate nuclei, evaluating also whether there was more texture information in 3D whole nuclei compared to a single 2D plane from the middle of the nucleus. RESULTS: All texture features showed the expected invariance properties. Almost all features were sensitive to variations in the nucleolar size and to the degree of margination of chromatin. Fourteen texture features from different categories had high discriminative power for separating the benign and malignant nuclei. The discrete texture features performed less than expected. There was more information on nuclear texture in 3D than in 2D. CONCLUSION: A set of 35 3D nuclear texture features was used successfully to assess nuclear chromatin patterns in 3D images obtained by confocal laser scanning microscopy, and as a proof of principle we showed that these features may be clinically useful for analysis of prostate neoplasia. PMID- 16373967 TI - Usefulness of immunological detection of the human telomerase reverse transcriptase. AB - OBJECTIVE: Recent years have seen a considerable wealth of studies conducted on the potential usefulness of telomerase determination in diagnosis, prognosis and targeted cancer therapy. The frequently used Telomeric Repeat Amplification Protocol assay suffers from some drawbacks, the most important being the rate of false positives. In situ analysis using well characterised antibodies directed against the human telomerase reverse transcriptase (hTERT) would therefore appear to be important to morphologically identify the nature of telomerase positive cells. METHODS: We performed immunostaining in a series of cultured cells and in normal, preneoplastic and tumour tissues from different organs using a monoclonal antibody directed against the catalytic subunit of telomerase. RESULTS: Immunoreactivity was not observed in perennial cells of terminally differentiated cardiac and skeletal muscular tissues or in small pyramidal cells of the cerebral cortex. Conversely, it was found in other normal somatic tissues as well as in precancerous lesions and in all tumour histotypes. CONCLUSIONS: Immunohistochemistry with a well characterised hTERT-specific monoclonal antibody permitted the identification of hTERT immunopositive cells in normal somatic tissues. Whether hTERT protein detected by immunostaining with hTERT-specific Tel 3 36-10 antibody is actually the degraded form of the protein that retains hTERT antigenicity but not enzymatic function, or whether it represents the real, potentially functional catalytic subunit of the enzyme, immunohistochemistry would not seem to represent a useful tool to investigate the role of telomerase and the mechanisms involved in its regulation. PMID- 16373968 TI - Ki67 predicts progression in early CIN: validation of a multivariate progression risk model. PMID- 16373970 TI - DNA ploidy and chromosome (FISH) pattern analysis of peripheral nerve sheath tumors. PMID- 16373972 TI - KRAS transitions and villous growth in colorectal adenomas. PMID- 16373974 TI - Geographic variation in surgical treatment for work-related carpal tunnel syndrome: does improved return to work matter? AB - Medical treatment of compensated work-related conditions has two objectives- improve injured workers' health status and allow safe and sustained return to work (RTW). Theoretically, the choice of treatment method should be based primarily on these objectives. Surgical treatment of work-related carpal tunnel syndrome (CTS) provides an opportunity to evaluate whether this occurs. The traditional method of open release has been complemented by an endoscopic procedure, particularly useful in work-related cases due to the anticipated benefit of earlier RTW. The objective of this study was to investigate the differences in surgical treatment for work-related CTS across eight US workers' compensation (WC) jurisdictions, and the factors associated with these differences. From all WC claims reported to a single insurer during the 1995-1999 period, we identified individuals with a one or two surgical procedures for work related CTS. Among selected individuals (n=4,421), about 20% were treated using the endoscopic procedure; this percentage had a ten-fold variation across the eight jurisdictions. However, utilization of endoscopic release did not increase during the study period, despite reports of better RTW outcomes. The highly jurisdictional nature of the US WC system, with significant differences in reimbursement levels for endoscopic procedures, and geographical differences in medical training were among the potential contributors to the observed variation in utilization. PMID- 16373975 TI - Ergonomic recommendations and their impact on child care workers' health. AB - The purpose of this study was to determine the impact ergonomics has had on the occurrence of musculoskeletal symptoms and injuries among child care workers in Wisconsin using a Classroom and Work Methods Survey developed for this investigation. Information on perceived need for and knowledge of ergonomic interventions, the extent to which ergonomic interventions had been introduced to child care settings, and barriers to implementation of ergonomic recommendations was gathered. Two hundred and fifty-eight respondents completed the survey. The majority of respondents was aware of most (18 of 20) of the recommended ergonomic interventions for child care centers and did not perceive a need to make ergonomic changes. Although the majority of respondents had implemented 15 out of the 20 recommendations, they were currently experiencing musculoskeletal pain while performing their jobs. The results of this study highlight the need for further analysis of specific child care tasks and identification of more effective methods to reduce the risk for musculoskeletal pain and injuries. PMID- 16373976 TI - Gender-age environmental associates of middle school students' low back pain. AB - PURPOSE: The purpose of this study was to describe adolescents' low back pain, investigate its prevalence and explore its associations with backpack load, time spent carrying loads, time sitting, and time playing sports, to determine whether relationships exist among these variables. METHOD: One hundred urban middle school students in Manchester, New Hampshire completed a questionnaire (55% response rate). Participants were between 13-14 years old. Weights of students, loaded backpacks, backpack contents, and students' heights were measured separately. RESULTS: Eighty-nine percent of the participants wore the two-straps backpack. Over eighty percent of them preferred to carry the load over two shoulders. The average load weighed 4.9 kg (approximating 9.6% of the participants' body weight). There was a significant association found between backpack carrying time and adolescent low back pain. CONCLUSION: Daily backpack carrying is a frequent cause of musculoskeletal discomfort for adolescents. The association between backpack carrying time and low back pain may provide the impetus for parents, teachers, and school administrators to decrease the prolonged carrying of backpacks. Further investigations should be conducted to evaluate the intervention effectiveness after implementation of controls. PMID- 16373977 TI - Fire-fighters' perspectives of the accuracy of the Physical Aptitude Test (P.A.T) as a pre-employment assessment. AB - Pre-employment assessments must accurately simulate job tasks and demands and select appropriate personnel to be considered effective. This study focussed on the perception of NSW fire-fighters in relation to the validity of the NSW Fire Brigade's pre-employment assessment, the Physical Aptitude Test. A qualitative method was used to gain a precise understanding of fire fighters' opinions of the accuracy of the Physical Aptitude Test. Information letters and consent forms were sent to an urban fire station with interested participants replying via the university. Six participants, who met the inclusion criteria were randomly selected for the study and in-depth, ethnographic, semi-structured interviews were conducted. The fire fighters believed in order for the Physical Aptitude Test to accurately simulate job demands and select the most capable fire fighters', it needed to be more physically demanding. However, participants believed that the "work-simulating" nature of the Physical Aptitude Test provided an accurate indication of the job tasks. PMID- 16373978 TI - Measuring empowerment in working life: a review. AB - This study identifies and describes questionnaires that measure empowerment in working life. Theoretical bases and empirical examination of the questionnaires are also reported. Nine questionnaires emerged from a database search including AMED, CINAHL, ERIC, MEDLINE and PSYCINFO. The main target groups were employees in general. Most authors share the same theoretical basis. Most of the questionnaires focus on intra-individual issues, while a smaller number deal with the interaction between individual and organization. Control and competence are frequently used dimensions. Cronbach's alpha for complete questionnaires ranged between 0.62 and 0.96. No comparisons with outcome of health were reported. Spreitzer's questionnaire has undergone the most comprehensive investigation. Research is required to achieve better understanding of the interplay between conditions at work and empowerment and health. PMID- 16373979 TI - Quality of life among unemployed and employed people in northern Sweden. Are there any differences? AB - This study analysed self-assessed quality of life (QoL), using a QoL questionnaire (Hornquist's QLcs) covering the life spheres, somatic health, mental well-being, cognitive ability, social and family life, activity, financial situation, meaning in life and a global score "entire life", in 487 unemployed subjects and 2917 employed subjects aged 25-64, in a population-based cross sectional study in northern Sweden. In line with previous findings, results showed that unemployed people exhibited poorer QoL. Unemployed women scored higher in existential life domains than unemployed men did. Unemployed men were worst off in terms of general life situation. Employed respondents benefited in QoL by a university/college education, while unemployed respondents with a university/college education did not. Close social relations and money reserve were associated with higher QoL generally. It was concluded that further research is needed to differentiate various aspects of QoL and unemployment, and to compare with other samples. PMID- 16373980 TI - Workers' compensation for psychological injury: demographic and work-related correlates. AB - This study aimed to determine the demographic and work-related factors that contributed to the filing of a workers' compensation claim for psychological injury. Four groups of employees were compared: those who filed a workers' compensation claim, those who sought psychological treatment for occupational stress but did not file a workers' compensation claim, those who experienced elevated stress levels but did not seek help or lodge a claim, and those who experienced stressful events at work but did not develop symptomatology. The results indicated few variables that could adequately explain why some stressed employees opted for a workers' compensation claim. The implications of these results are discussed. PMID- 16373981 TI - Employment transitions for clubhouse members. AB - Using a longitudinal dataset which followed 2195 individuals employed in 3379 separate job placements over a four-year period, this paper explores movement between the employment supports, [Transitional (TE), Supported (SE), and Independent Employment (IE)], offered by clubhouses. Sixty-four percent of employed members held only one job (N=1395) and 36% held multiple jobs during the study (N=791). Patterns of movement were consistent for transitions between the first and second job and subsequent transitions. Forty-six percent of individuals holding multiple jobs moved from one employment type to another. When movement occurred clubhouse members were significantly more likely to move from employment types offering more supports to those that offer less supports. PMID- 16373982 TI - Factors affecting the organizational responses of employers to workers with injuries. AB - The organizational responses of employers to work-related injuries is one of several significant influences on return-to-work outcomes. Thus, understanding the factors that lead to better or worse organizational responses to work injuries may ultimately help to improve success in this area. The purpose of this study was to systematically explore factors that might influence the organizational responses of employers to injured workers, based on employee perceptions. Cross-sectional survey data were collected from 2,943 subjects with work-related injuries which had occurred less than eight weeks prior to survey completion. Measured variables included pre-injury demographic and job factors, injury circumstances, and a measure of post-injury events that comprised the organizational response. Multivariate linear regression results show that age, gender, job dissatisfaction before injury, prior difficulty performing job tasks, injury severity, back injury and lost time were all associated with negative organizational responses, suggesting potential opportunities for intervention. PMID- 16373983 TI - Single subject research methods in rehabilitation. AB - The article describes single-subject research designs as an efficient and cost effective way to assess the impact of targeted interventions on individual behavior. The authors overview the basic tenets of single-subject research, provide examples of several types of single-subject designs, and describe the utility of single-subject designs in rehabilitation research. PMID- 16373984 TI - Crisis and opportunity: coaching older workers in the workplace. PMID- 16373985 TI - [Investigation of Energy Subtraction in a CT System.]. AB - Developments in CT units for medical diagnoses permit to energy subtraction in CT radiography. Experiments were done for CT energy subtraction using a TOS phantom, self-made phantom and dry human skull at two tube voltages of 80kV and 135kV. The TOS phantom filled with water had six compartments of air, polyoxymethylene, polymethyl methacrylate, polyamide and polypropylene. The self-made phantom which included a cylinder filled with contrast medium was used for the contrast vessel. Bone contents disappeared selectively from images of both the self-made phantom and the human skull. In these cases, disappearance of selected materials was achieved by changing the coefficient of CT subtraction. It was confirmed that the procedure could be used for a variety of materials. PMID- 16373986 TI - [Clinical calibration dosimetry in JSMP-01: measurements using Farmer-type cylindrical ion chambers.]. AB - The Japan Society of Medical Physics (JSMP) Task Group published Standard dosimetry of absorbed dose in external beam radiotherapy (Standard dosimetry 01) as a new high-energy photon and electron dosimetry protocol in 2002. In this study, we present Standard dosimetry 01 as the JSMP-01 protocol for the convenience of users. This protocol is based on using an ion chamber having a (60)Co absorbed dose to water calibration coefficient, N(D,w), which is calculated from a (60)Co exposure calibration coefficient, N(c). We present dose comparisons between a reference chamber and various Farmer-type cylindrical chambers with different wall materials. The absorbed dose to water was compared at the calibration depths of 5 cm for a (60)Co beam, 10 cm for photons, and d(c) = 0.6 R(50) - 0.1 (cm) for electrons according to JSMP-01. The JARP chamber in the Kyushu Regional Center which meets third-order standards in Japan was used as the reference chamber. The absorbed dose to water for the Farmer-type chambers determined according to JSMP-01 agreed with that for the JARP chamber within 1% for photon and electron beams. The doses obtained by JSMP-01 and the Japan Association of Radiological Physics protocol (JARP-86) were also compared for photon and electron beams. For the Farmer-type chambers with photon beams, JSMP 01 results were up to 1.5% higher than JARP-86 results. For electron beams JSMP 01 results were higher than JARP-86 results by 1.3-2.8%. PMID- 16373987 TI - [Clinical calibration dosimetry in JSMP-01: measurements using plane-parallel ion chambers.]. AB - The Japan Society of Medical Physics (JSMP) Task Group published Standard dosimetry of absorbed dose in external beam radiotherapy (Standard dosimetry 01) as a new high-energy photon and electron dosimetry protocol in 2002. In this study, we present Standard dosimetry 01 as the JSMP-01 protocol for the convenience of users. This protocol is based on using an ion chamber having a (60)Co absorbed dose to water calibration coefficient, N(D,w), which is calculated from a (60)Co exposure calibration coefficient, N(c). We present dose comparisons between a reference chamber and various plane-parallel chambers. The absorbed dose to water was compared at the calibration depth of 5 cm for a (60)Co beam and d(c) = 0.6R(50) - 0.1 (cm) for electron beams according to JSMP-01. The absorbed dose to water calibration coefficients, [N(D,w)](Co) and [N(D,w)](18E), for the plane-parallel chambers were also determined by (60)Co and electron beam cross-calibrations using a reference chamber. The dose for the plane-parallel chambers derived from [N(D,w)](Co) and [N(D,w)](18E) was compared to that for the reference chamber using electron beams. The JARP chamber in the Kyushu Regional Center which meets third-order standards in Japan was used as the reference chamber. The doses for the plane-parallel chambers determined according to JSMP 01 agreed with that for the JARP chamber within 1% and 2% for (60)Co and electron beams, respectively. For electron beams, the doses for the plane-parallel chambers calculated from [N(D,w)](Co) and [N(D,w)](18E) were within 1.5% and 1.0% compared to those for the JARP chamber, respectively, except for the Exradin A10 chamber. PMID- 16373988 TI - Structural inhomogeneity in mammography quality control phantoms detected by refraction-enhanced synchrotron radiation imaging. AB - Synchrotron radiation imaging with the refraction-enhancement mode visualized structural inhomogeneities in phantoms used for image quality control of mammography. Eight phantoms were examined, all of which were manufactured in the United States and approved by the American College of Radiology as dedicated phantoms. In addition to fiber- and mass-mimicking test objects, each phantom has 5 groups of calcification specks of various sizes. Synchrotron radiation (SR) imaging was performed at Spring-8, a synchrotron radiation facility in Japan. Images were obtained with monochromatic 20-keV x-ray beams, a radiation field of 15 mm X 26 mm at a sample plane, a CCD camera with a resolution of 6 micrometers as a detector, and a sample-to-detector distance of 10 to 11 m. Two hundred and forty specks were evaluated in total in the SR images, and the surrounding area of each speck was also included. Evaluation of the images showed that 14 crack like structures were depicted near specks, and there were 62 specks with attached void(s) or air bubble(s). Refraction-enhanced SR imaging sensitively detected structural inhomogeneities and abnormalities in phantoms which were implicitly agreed to have a homogeneous matrix and test objects without foreign substances. A possible manufacturing-dependent quality issue was identified. The effect of inhomogeneities detected by SR imaging on visual scoring of specks could not be identified in the tested phantoms; this should be assessed on images of other phantoms in a future study. PMID- 16373989 TI - Validation of the blurring of a small object on CT images calculated on the basis of three-dimensional spatial resolution. AB - We determine three-dimensional (3D) blurring of a small object on computed tomography (CT) images calculated on the basis of 3D spatial resolution. The images were characterized by point spread function (PSF), line spread function (LSF) and slice sensitivity profile (SSP). In advance, we systematically arranged expressions in the model for the imaging system to calculate 3D images under various conditions of spatial resolution. As a small object, we made a blood vessel phantom in which the direction of the vessel was not parallel to either the xy scan-plane or the z-axis perpendicular to the scan-plane. Therefore, when scanning the phantom, non-sharpness must be induced in all axes of the image. To predict the image blurring of the phantom, 3D spatial resolution is essential. The LSF and SSP were measured on our scanner, and two-dimensional (2D) PSF in the scan-plane was derived from the LSF by solving an integral equation. We obtained 3D images by convoluting the 3D object-function of the phantom with both 2D PSF and SSP, corresponding to the 3D convolution. Calculated images showed good agreement with scanned images. Our technique of determining 3D blurring offers an accuracy advantage in 3D shape (size) and density measurements of small objects. PMID- 16373990 TI - Immune-enhancing role of vitamin C and zinc and effect on clinical conditions. AB - Vitamin C concentrations in the plasma and leukocytes rapidly decline during infections and stress. Supplementation of vitamin C was found to improve components of the human immune system such as antimicrobial and natural killer cell activities, lymphocyte proliferation, chemotaxis, and delayed-type hypersensitivity. Vitamin C contributes to maintaining the redox integrity of cells and thereby protects them against reactive oxygen species generated during the respiratory burst and in the inflammatory response. Likewise, zinc undernutrition or deficiency was shown to impair cellular mediators of innate immunity such as phagocytosis, natural killer cell activity, and the generation of oxidative burst. Therefore, both nutrients play important roles in immune function and the modulation of host resistance to infectious agents, reducing the risk, severity, and duration of infectious diseases. This is of special importance in populations in which insufficient intake of these nutrients is prevalent. In the developing world, this is the case in low- and middle-income countries, but also in subpopulations in industrialized countries, e.g. in the elderly. A large number of randomized controlled intervention trials with intakes of up to 1 g of vitamin C and up to 30 mg of zinc are available. These trials document that adequate intakes of vitamin C and zinc ameliorate symptoms and shorten the duration of respiratory tract infections including the common cold. Furthermore, vitamin C and zinc reduce the incidence and improve the outcome of pneumonia, malaria, and diarrhea infections, especially in children in developing countries. PMID- 16373991 TI - Erythrocyte membrane phospholipid composition as a biomarker of dietary fat. AB - BACKGROUND/AIMS: In a cross-sectional study, we investigated the relationship between erythrocyte membrane phospholipid fatty acid composition and dietary fat; we also investigated roles of menopausal status, age, body mass index (BMI) and waist-to-hip ratio (WHR) in interindividual variation of the biomarker. METHODS: Study participants were 204 women, aged 39-65 years, drawn from the ORDET cohort and selected as controls in a study of breast cancer. Membrane composition was assessed using capillary gas chromatography. Dietary fat composition was evaluated using a food frequency questionnaire. RESULTS: In pre- and postmenopausal women, erythrocyte membrane phospholipid levels of linoleic acid, oleic acid, and mono-unsaturated fatty acids were significantly associated with corresponding dietary measures (partial correlation coefficients: 0.23 and 0.39; 0.45 and 0.47; 0.40 and 0.48; respectively, in pre- and postmenopausal women). Among postmenopausal women, membrane poly-unsaturated fatty acids were correlated with the corresponding dietary measure (r=0.39, p<0.001). Membrane eicosapentanoic and docosahexanoic acid levels were significantly correlated with intake of fish/shell fish : r=0.21 and r=0.43 (premenopausal), and r=0.41 and r=0.44 (postmenopausal). Age, BMI and WHR had independent effects on membrane lipid composition. Age was associated with delta-6 desaturase activity in postmenopausal women (r=0.25, p<0.05). BMI was negatively associated with delta-9 desaturase activity in both pre- and postmenopausal women (r=-0.29, p=0.01 and r= 0.22, p<0.01, respectively). WHR was negatively associated with delta-5 desaturase activity in pre-menopausal women (r=-024, p<0.05). CONCLUSIONS: Erythrocyte membrane levels of some specific fatty acids can be used as biomarkers of these fatty acids as proportions of dietary fat. PMID- 16373992 TI - Assessment of cardiovascular risk factors among Hungarian university students in Budapest. AB - BACKGROUND/AIMS: The lack of data regarding cardiovascular risk factors among university students in Hungary prompted us to undertake a cross-sectional pilot study in students at Semmelweis University, Budapest. METHODS: A total of 264 students (78 males, mean age 21.4+/-2.6 years, and 186 females, mean age 21.2+/ 2.6 years) participated in the study. Anthropometric measures, blood pressure and serum lipid parameters were investigated. The relationship between body mass index and cardiovascular disease risk factors were analyzed. RESULTS: The occurrence of overweight and abdominal obesity was significantly higher in the male group. The mean serum lipid values were within the acceptable range for both genders; however, in a significantly greater number of male than female students the HDL-cholesterol concentration (<1 mmol/l) was in the low range. High lipoprotein(a) values were seen in 33% of the male and 22.6% of the female students. In both groups of students the body mass index exhibited a positive correlation with waist circumference, total cholesterol, LDL-cholesterol, apolipoprotein B (p<0.01), and systolic and diastolic blood pressure. In the female students there was an inverse correlation between body mass index and HDL cholesterol concentrations. CONCLUSIONS: The anthropometric and lipid parameters of the student population studied were more favorable than those for an age matched Hungarian youth population. Nevertheless, the higher occurrence of elevated blood pressure, overweight, abdominal obesity, lower HDL-cholesterol in the male group should be regarded as a warning sign, all the more since their diet was imbalanced and their knowledge in this field was incomplete. The results of our preliminary study point out the importance of teaching nutrition more efficiently, not only during university training but also at an earlier age. PMID- 16373993 TI - Dose-dependent inhibition of the post-prandial glycaemic response to a standard carbohydrate meal following incorporation of alpha-cyclodextrin. AB - BACKGROUND: This study evaluated the dose-response effects of alpha-cyclodextrin, a cyclic oligosaccharide, on the glycaemic and insulinaemic responses to the consumption of a standard carbohydrate meal. METHODS: In a double-blind, randomised, cross-over design, 10 healthy subjects consumed boiled white rice containing 50 g of digestible carbohydrate to which 0 (control), 2, 5 or 10 g of alpha-cyclodextrin was added. Plasma glucose and insulin concentrations were determined prior to and for 2 h after consumption of each meal. RESULTS: The area under the plasma glucose curve was negatively related to the dose of alpha cyclodextrin (r(2)=0.97, p=0.02), with the areas being significantly reduced at the 5- and 10-gram doses compared with the control (p<0.05). alpha-Cyclodextrin did not affect the area under the plasma insulin curve (p=0.39). Higher doses of alpha-cyclodextrin resulted in greater satiety, but were associated with reduced palatability and an increased incidence of minor gastrointestinal complaints (stomach ache, nausea, bloating). CONCLUSION: alpha-Cyclodextrin reduces the glycaemic response to a standard carbohydrate meal in a dose-dependent manner and may be useful as an ingredient for reducing the glycaemic impact of such foods. PMID- 16373994 TI - Relationship between insulin resistance and lipid peroxidation and antioxidant vitamins in hypercholesterolemic patients. AB - BACKGROUND: Several studies have reported that insulin resistance and compensatory hyperinsulinemia increased lipid peroxidation, suggesting the linking to each other. We investigated the relationships between insulin resistance index HOMA-IR and lipid peroxidation, plasma antioxidant status in non diabetic, hypercholesterolemic patients. METHODS: We measured the urinary excretion of 8-epi-prostaglandin F(2)(alpha)(PGF(2)(alpha)) levels as a measure of lipid peroxidation in vivo, total radical trapping antioxidant potential (TRAP) and fat-soluble antioxidant vitamins in 76 non-diabetic subjects with hypercholesterolemia (mean age 59 years, 25 males and 51 females). Insulin resistance was evaluated by homeostasis model assessment (HOMA-IR) derived from fasting glucose and insulin concentrations. RESULTS: HOMA-IR was positively correlated with the urinary excretion of PGF(2)(alpha) (r=0.222, p<0.05) and negatively correlated with the TRAP (r=-0.211, p<0.05) in total subjects. Furthermore, there were significant inverse relationships between HOMA-IR and lipid corrected fat-soluble vitamins such as beta-carotene (r=-0.297, p<0.01) and gamma-tocopherol (r=-0.243, p<0.05) and also significant inverse relation was found between lipid corrected beta-carotene and the urinary PGF(2)(alpha)excretion (r=-0.205, p<0.05). When total subjects were divided into three groups according to tertiles of HOMA-IR, significant differences in urinary PGF(2)(alpha)excretion (p<0.05) and lipid corrected beta-carotene (p<0.005) among the three groups were observed. The highest HOMA-IR group had the higher levels of urinary PGF(2)(alpha)excretion and lower levels of plasma beta-carotene compared with the lowest HOMA-IR group. CONCLUSION: Our data showed that the insulin resistance of hypercholesterolemic patients increased oxidative stress and negatively influenced plasma antioxidant system. These results provide evidence in understanding mechanism linking insulin resistance and oxidative stress accompanied by reduced antioxidant system. PMID- 16373995 TI - Ethylene glycol poisoning in the Czech Republic (2000-2002). AB - BACKGROUND: The objective of the study was to evaluate the severity of kidney damage retrospectively and to analyze prognostic factors following ethylene glycol (EG) poisonings. METHODS: Data concerning the clinical course of patients with EG poisoning between 2000 and 2002 were analyzed. The chi2 test, Student's t test, Fisher's test and the calculation of linear correlation coefficients were used for statistical analysis. RESULTS: Thirty-three discharge records were obtained. Three patients died, and 11 patients developed acute renal failure (mean maximum serum creatinine level 618 micromol/l). Upon discharge, serum creatinine levels were still elevated in 10 patients. In all but 1 patient, renal function completely normalized within 20 months after intoxication. CONCLUSION: Adult men are the most endangered segment of the population. The most important prognostic factors are the ingested dose and early antidotal treatment. EG toxic kidney damage is reversible. PMID- 16373996 TI - Telomere shortening and haemodialysis. AB - BACKGROUND: Increased oxidative stress is a well described feature of haemodialysis (HD). This is secondary to an increase in the production of reactive oxygen species and impaired antioxidant mechanisms. Telomeres are the specialized ends of eukaryotic chromosomes and consist of tandemly repeated DNA sequences. Telomeres shorten with each cell division and it is well known that telomere length in peripheral blood mononuclear cells (PBMCs) decreases with age. Telomere shortening rate is increased by oxidative stress. In this study we have examined a possible relationship between oxidative stress and telomere shortening in haemodialysis. METHODS: 20 control subjects, 20 non-diabetic and 18 diabetic HD patients were studied. Peripheral blood mononuclear cell telomere length, plasma malondialdehyde plus 4-hydroxyalkenal (MDA+4-HAE) concentration (a marker of oxidative stress) and C-reactive protein (CRP) concentration were measured. RESULTS: MDA+4-HAE and CRP were significantly higher in the HD patients (CRP, controls 7.5 +/- 1.5, HD patients 16.4 +/- 3.1 mg/l, p < 0.05). There was no difference in mean telomere length between the HD patients and controls (control, 8,283 +/- 179 bp; non-diabetic HD, 7,966 +/- 160 bp; diabetic HD, 8,033 +/- 197 bp) but age adjusted residual telomere length was inversely associated with the length of time on dialysis (r = -0.35, p = 0.03). CONCLUSION: These results suggest that length of time on dialysis is independently associated with increased telomere shortening in HD patients. We hypothesise that this reflects cumulative DNA exposure to oxidative stress. PMID- 16373997 TI - Outcome of autogenous fistula construction in hemodialyzed patients over 75 years of age. AB - BACKGROUND: There are controversies regarding the feasibility of autogenous vascular access creation in elderly hemodialysis (HD) patients. The aim of this retrospective study was to evaluate the results of creating different types of autogenous arteriovenous fistulas (AVFs) in a consecutive series of HD patients over 75 years of age. METHODS: The analysis was performed in 131 patients (65 females, 66 males, average age 79.1 +/- 3.6 years) in whom the creation of an autogenous AVF was considered within a 6-year period (February 1998 to February 2004). Among them, 26.7%were diabetics, 66.3% had hypertension, 30.7% were smokers, and 35.6% were obese. Patient survival and primary and secondary AVF patency were assessed. RESULTS: The survival rates for patients were 94, 88, 66, and 45% at 6 months and at 1, 3, and 5 years, respectively. Successful autogenous AVF formation was finally achieved in 107 patients (81.6%): in 99 patients in the forearm and in 8in the upper arm. A Kaplan-Meier analysis showed primary AVF patency rates of: 74 +/- 4.3% (+/- SE) at 1 month; 70 +/- 4.7% at 6 months; 59 +/ 4.9% at 1 year; 59 +/- 4.9% at 2 years; 59+/- 4.9% at 3 years; 59 +/- 4.9% at 4 years, and 58 +/- 4.9% at 5 years. The secondary patency rates were: 95 +/- 2.0; 92 +/- 2.2; 84 +/- 3.3; 79 +/- 4.0; 72 +/- 4.3; 71 +/- 4.4, and 69 +/- 4.5% in the corresponding periods, respectively. All postoperative complications in 10 patients were treated surgically, if applicable, without endovascular techniques. CONCLUSIONS: By exploiting all suitable types of autogenous AVF it is possible to establish the best form of vascular access even in the majority of elderly patients. PMID- 16373998 TI - Imbalance between detached circulating endothelial cells and endothelial progenitor cells in chronic kidney disease. AB - BACKGROUND: Patients with chronic kidney disease (CKD) display endothelial dysfunction and are at a high risk for atherosclerotic cardiovascular disease (CVD). Recent studies suggest that circulating detached endothelial cells (CECs) and stimulated endothelial progenitor cells (EPCs) from the bone marrow may reflect endothelial damage. METHODS: We correlated the levels of CECs expressing the endothelial cell inflammation marker (MICA+ cells) and EPCs (Tie-2+ or VEGFR 2+ cells) in a population of 19 (55 +/- 3 years; 42% males) patients with advanced CKD (median glomerular filtration rate 8 ml/min). In addition, the levels of CD-31+ cells were investigated. Twenty healthy age- (49 +/- 2 years) and gender- (50% men) matched subjects served as controls. RESULTS: CECs expressing MICA were increased (7.6 +/- 2.7 vs. 1.6 +/- 0.3%; p < 0.05) in CKD patients, however EPCs expressing Tie-2 or VEGFR-2 were significantly decreased (0.16 +/- 0.07 vs. 0.53 +/- 0.15%; p < 0.05, and 0.42 +/- 0.10 vs. 2.80 +/- 0.72%; p < 0.01, respectively) as compared to controls. Furthermore, we also found that the levels of CD-31+ cells were significantly elevated (22.8 +/- 4.2 vs. 9.4 +/- 0.8%; p < 0.01) in CKD patients. Patients on angiotensin-converting enzyme (ACE) inhibitors tended (p = 0.06) to have higher levels of VEGFR-2+ cells (0.57 +/- 0.14 vs. 0.16 +/- 0.11%). CONCLUSION: Our results suggest that there is a marked imbalance between the CEC and EPC numbers in patients with CKD. Further research is needed to evaluate the independent role of inflammatory endothelial markers as well as the effects of ACE inhibitors on mobilization of EPCs in patients with advanced CKD. PMID- 16373999 TI - Reduced cytokine induction and removal of complement products with synthetic hemodialysis membranes. AB - The increasing use of high-flux membranes for hemodialysis (HD) has raised concerns that these membranes may confer a higher risk of exposure to cytokine inducing, bacterial substances (CIS) in the dialysate. Several studies, however, reported higher transfer of CIS through low-flux cellulosic than high-flux synthetic membranes. This surprising paradox was explained by adsorption of CIS to certain high-flux membranes. In order to investigate flux and membrane type independently, we studied two synthetic Polyflux (PF) membranes of the same type but with different flux properties and compared them to a cellulosic membrane (Cuprophan). Three different approaches were employed: (1) cytokine induction in whole blood during in vitro HD contaminated with bacterial filtrates, (2) removal of recombinant C5a, and (3) transfer of purified lipopolysaccharide (LPS). After 90 min recirculation of whole blood, the appearance of IL-6-inducing substances on the blood side was lowest with high-flux PF (1.1 +/- 0.2 ng/ml), slightly higher with low-flux PF (1.9 +/- 0.7 ng/ml) and highest with Cuprophan (4.1 +/- 1 ng/ml). Recombinant C5a added to plasma on the blood side was markedly removed by high-flux PF (by 83%), to a lesser degree and only in the presence of ultrafiltration with low-flux PF (by 54%) and not significantly with Cuprophan (by 11%). Significant transfer of purified LPS from the dialysate onto the blood side was only observed with the cellulosic membrane. We conclude that in contrast to cellulosic membranes, certain synthetic membranes do not permit transfer of LPS. Cytokine induction on the blood side is further reduced by the use of high flux membranes due to removal of activated complement factors. PMID- 16374000 TI - Normobaric hyperoxia - a promising approach to expand the time window for acute stroke treatment. PMID- 16374001 TI - The neurovascular unit and the key role of astrocytes in the regulation of cerebral blood flow. PMID- 16374002 TI - Isolated dorsal midbrain infarct: an uncommon cause of pure sensory stroke. PMID- 16374003 TI - Beauty parlor stroke syndrome. PMID- 16374004 TI - A giant intracavernous carotid artery aneurysm embedded in pituitary macroadenoma presenting with pituitary apoplexy. PMID- 16374005 TI - Cognitive assessment of the non-demented elderly community dwellers in Spain. AB - OBJECTIVE: The prevalence of cognitive impairment and population normative values for cognitive function were assessed, for the first time, in 2,630 subjects representative of the non-demented community dweller population > or =65 years in Spain, a southern European country with a sizable proportion of illiterate senior citizens. METHODS: Data were collected cross-sectionally by interview, using a structured questionnaire. Cognitive function was assessed using the Mini-Examen Cognoscitivo (MEC; Spanish-validated version of the Mini-Mental State Examination). RESULTS: A total of 22.4% of the subjects presented with cognitive impairment (MEC < or =22). The proportion of subjects with cognitive impairment was double in women versus men, rose to 46.2% in subjects aged > or =85 years and to 34.8% in subjects with no formal education, and was higher among those who had limitations in instrumental activities of daily living (p < 0.001). In subjects with no formal education, 25% registered mean MEC scores compatible with probable cognitive impairment. CONCLUSIONS: One of every 5 Spanish senior citizens presents with cognitive impairment, a proportion that increased among women, subjects with no formal education and with higher age. PMID- 16374006 TI - Cognitive impairment in patients with idiopathic normal pressure hydrocephalus. AB - Cognitive impairment in patients with idiopathic normal pressure hydrocephalous (iNPH) has not been clearly documented. We performed standardized neuropsychological assessments of 21 patients with iNPH and of 42 patients with Alzheimer's disease (AD) matched to the patients with iNPH 2:1 by age, sex, and Mini-Mental State Examination score. Compared with the AD group, the iNPH group scored significantly higher on the orientation subtest of the Alzheimer's Disease Assessment Scale and on the general memory and delayed recall subtests of the Wechsler Memory Scale-Revised (WMS-R), and significantly lower on the attention/concentration subtest of the WMS-R and on the digit span, arithmetic, block design and digit symbol substitution subtests of the Wechsler Adult Intelligence Scale-Revised. The impairment of frontal lobe functions is disproportionately severe and memory impairment is disproportionately mild in patients with iNPH compared with AD. Recognition of these features contributes to an early diagnosis, which can lead to a better prognosis. PMID- 16374007 TI - Immunohistologic attempt to find carcinogenesis from hepatic progenitor cell in hepatocellular carcinoma. AB - AIM: To clarify whether hepatocellular carcinoma (HCC) originates from hepatic progenitor cells and whether there is any correlation with the clinicopathologic factors of HCC, we reviewed 217 resected HCC specimens. METHODS: Immunohistochemical examination of cytokeratin (CK) 7, CK19, CD34, and CD117 (c KIT) was performed. Overexpression of CK7 and CK19 indicates differentiation from cholangiocellular and hepatic progenitor cells, while overexpression of CD34 and CD117 indicates hepatic stem cells. Fresh specimens were obtained from 20 HCC patients for mutation of the c-KIT gene. RESULTS: CK7, CK19, and CD117 were positive in 41, 9.7, and 0.9% of the HCC specimens, respectively, and CD34 was never positive. None of the fresh HCC specimens demonstrated a c-KIT mutation. CK19 positivity was significantly correlated with a positive hepatitis B core antibody, and with poor survival outcome, and tended to correlate with poor histologic differentiation. CONCLUSION: These results suggest that: (i) about 10% of HCCs with typical histologic features originate from an intermediate hepatic progenitor cell, such as the canal of Hering and oval cells in the rat, or acquire the characteristics of cholangiocellular epithelium by metaplasia; (ii) HCC with typical histologic features rarely originates from hepatic stem cells, and (iii) patients with CK19-positive HCC have a poor prognosis. PMID- 16374008 TI - Effect of anti-OprF-OprI immunoglobulin on APACHE II score in a porcine two-hit model of hemorrhagic shock/resuscitation and pseudomonas aeruginosa sepsis. AB - BACKGROUND: Up to now, randomized clinical trials of treatment of bacterial sepsis with immunoglobulins show conflicting results. This paper investigates the effect of prophylactic immunization with anti-OprF-OprI antiserum on the APACHE II score in a clinically relevant two-hit model of hemorrhagic shock/resuscitation followed by Pseudomonas aeruginosa sepsis in pigs. METHODS: Twenty-three German Landrace-Hybrid pigs underwent chronic implantation of vascular catheters (internal and external jugular vein, carotic and pulmonary artery), hemorrhagic shock (mean blood loss 40% of estimated blood volume) for 45 min, followed by resuscitation with crystalloid, colloid, and shed blood. Randomization was to a control group (no immunization, n=6), an F-I group (50 mg/kg i.p. anti-OprF-OprI immunoglobulin, n=6), an S group (50 mg/kg i.p. unspecific porcine immunoglobulins, n=6), and a PS group (50 mg/kg i.p. immunoglobulin against the antigens of heat-killed P. aeruginosa, n=5). After at least 18 h for recovery from anesthesia, the pigs underwent a continuous intravenous infusion of P. aeruginosa for 48 h. Thereafter, the animals were monitored for another 48 h and then dissected. RESULTS: The APACHE II score significantly increased from baseline value in all groups during bacterial challenge. However, there were no between-group differences in APACHE II score. In contrast, pigs of the F-I and PS groups showed significant lower lung concentrations of P. aeruginosa (p<0.05 vs. control group) at autopsy. CONCLUSION: These experimental data suggest that under comparable clinical conditions, a prophylactic immunization with anti-OprF-OprI immunoglobulin would not have an overall benefit to patients with P. aeruginosa sepsis. PMID- 16374009 TI - Neutrophil elastase inhibitor ameliorates reperfusion injury in a canine model of lung transplantation. AB - BACKGROUND: We investigated the effects of neutrophil elastase inhibitor ONO-5046 Na on lung ischemia-reperfusion injury in a canine model of single lung transplantation. METHODS: 24 mongrel dogs, 12 donors and 12 recipients, were used for single lung transplantation. Lung grafts were preserved for 18 h by cold ischemia then transplanted into the left thoracic cavity of recipients. In 6 recipients (ONO group), a bolus of ONO-5046 Na (10 mg/kg) was introduced before reperfusion and followed by continuous infusion (10 mg/kg/h). The remaining 6 recipients (control group) did not receive ONO-5046 Na and thus served as controls. We evaluated lung function and respiratory parameters over 240 min. RESULTS: The total cell number in bronchoalveolar lavage fluid increased significantly in the control group in comparison to that in the ONO group. Histologic scores after 4 h of reperfusion and myeloperoxidase activity were significantly lower in the ONO group than in the control group. CONCLUSION: Neutrophil elastase inhibitor ONO-5046 Na may be useful in ameliorating lung reperfusion injury after transplantation. PMID- 16374010 TI - Expression of CD97 and adhesion molecules on circulating leukocytes in patients undergoing coronary artery bypass surgery. AB - OBJECTIVE: Leukocyte activation is thought to be responsible for the adverse effects and postoperative complications following cardiopulmonary bypass (CPB). A novel cell surface molecule, CD97, is a sensitive marker of leukocyte and primary lymphocyte activation. The present study aimed to determine the activation of different leukocyte subsets by comparing the expression of CD97 and adhesion molecules (CD11, CD18) in patients receiving coronary surgery with or without CPB. METHODS: 30 patients were enrolled and scheduled for coronary bypass surgery under CPB (20 patients, group A) and with off-pump (OP) operation (10 patients, group B). Blood samples were taken before and during surgery, and over the following first week. RESULTS: Here, we report an early decrease in CD97 expression of granulocytes (PMN) and monocytes (MC) followed by an intensive increase reaching the maximum on postoperative days 2 and 3 in patients operated with CPB. The rate of active CD97-positive lymphocytes showed a marked, gradual increase until postoperative day 3 and remained elevated up to day 7 after CPB. OP surgery resulted in moderate alteration in the presence of CD97 on PMN, MC and lymphocytes. The expression of adhesion molecules was similar to CD97 in all leukocyte subsets. CONCLUSION: The findings about CD97 expression suggest considerable leukocyte activation following coronary bypass with CPB compared to OP surgery. The collected data show that the lymphocytes are highly activated and involved in leukocyte sequestration after CPB. Moreover, the importance of CD97 in CPB-related inflammatory response can be stated. PMID- 16374011 TI - Kupffer cells infiltrate liver tissue early after ischemia-reperfusion and partial hepatectomy. AB - Kupffer cells, ED2+macrophages of the liver, play an important role in liver damage and regeneration. It is proposed that Kupffer cells are stationary and regenerate after acute liver trauma by local proliferation. We analyzed their kinetics in three surgically relevant murine models of acute liver injury: partial liver resection, ischemia with reperfusion and sepsis. We found an early increase in ED2+cells after 0.5 h and a maximum after 12 h. These results suggest an infiltration of the cells early after the injury and a later local proliferation. These ED2+macrophages are localized predominantly periportally; nearly no macrophages are found pericentrally, except in the sepsis model. Therefore, a shifting of macrophages from portal to central seems to be unlikely, suggesting a hepatic zonation of homing factors. PMID- 16374012 TI - New surgical technique for portal venous port system in Swine. AB - The need for serial blood sampling and continuous infusion using the portal vein for experimental surgery and pharmacological studies is evident. As suitable animal models are rare, we established a new technique to implant a permanent vascular access device to the portal vein in swine. All operations in study animals (n=8) were technically feasible due to a regularly detected side branch of the portal vein within the inferior pancreatic region; complications did not occur. Mean operation time was 48+/-17 min. This study clearly shows that the portal venous access is feasible and can be achieved safely using the inferior pancreatic vein for a permanent vascular access port system. PMID- 16374013 TI - Pitavastatin prevents bacterial translocation after nonpulsatile/low-pressure blood flow in early atherosclerotic rat: inhibition of small intestine inducible nitric oxide synthase. AB - BACKGROUND: Cardiopulmonary bypass decreases intestinal mucosal blood flow because of nonpulsatile and low-pressure blood flow resulting in bacterial translocation (BT) and atherosclerosis also has peripheral blood flow deficiency. The risk of nonpulsatile and low-pressure blood flow for atherosclerotic animals and the effect of statin administration, which has pleiotropic effects, were studied. METHODS: Wistar rats were divided into four groups: group N (normal diet), group C (high-cholesterol diet), group S (group C plus pitavastatin therapy), and group I [group C plus inducible nitric oxide (iNOS) inhibitor therapy]. First of all, vascular responses were measured. Then the rats underwent nonpulsatile/low-pressure blood flow in the intestine, and the serum peptidoglycan concentration as a parameter of BT, the small intestinal PO(2) ratio (intestinal PO(2)/PaO(2)) as a parameter of mucosal blood flow, and NO concentrations were measured before surgery (T0), at the end of 90 min of stenosis (T1), and 90 min after the release of stenosis (T2). Immunostaining for nitrotyrosine was also performed at T2. RESULTS: Group C had vascular endothelial dysfunction without histological changes, which indicated early atherosclerosis. The serum peptidoglycan concentration increased significantly at T2 only in group C. The intestinal PO(2) ratio was decreased at T1 in all the groups, and retuned to baseline at T2 in group N and group S, but not in group C or group I. Jejunal NO only in group C was significantly higher at all time points and ileal NO production at T1 and T2. There tended to be a positive stain for nitrotyrosine along the mucosal epithelium in group C. CONCLUSION: In the setting of early atherosclerosis, intestinal blood flow does not only improve after nonpulsatile/low-pressure blood flow but causes BT because of a large amount of NO from high enzymatic intestinal iNOS activity, and pitavastatin treatment can prevent BT by improving both issues. PMID- 16374014 TI - Radiofrequency ablation using perfused needle electrodes - study of intermittent and continuous triple needle application ex vivo. AB - To increase necrotic zones, bovine livers were treated by means of three parallel oriented radiofrequency ablation (RFA) needles spaced at 3 cm using a puncture guide. The triple application was varied as a continuous and intermittent energy application compared to a single needle applicator. In all three study arms the applied energy (60 W) and the perfusion rate (240 ml/h) were kept constant. After treatment the smallest necrosis diameter was determined. In addition, temperature and the device's power output were monitored. Our study shows that synchronous use of three RFA application needles achieves significantly larger necrosis zones ex vivo than does single needle application. Intermittent energy application heats up the necrosis faster and more evenly with highest average temperature than continuous energy application. PMID- 16374015 TI - Stapled haemorrhoidopexy for haemorrhoids in combination with lateral internal sphincterotomy for fissure-in-ano. AB - The aim of this prospective study is to describe the combined technique and results of stapled haemorrhoidopexy and lateral internal sphincterotomy for patients suffering from prolapsing 3rd-degree haemorrhoids and chronic fissure-in ano. During the period from 1999 to 2004, 26 patients underwent combined surgical treatment for anal fissure and prolapsing symptomatic haemorrhoids. Preoperative and postoperative clinical evaluation and the patient's degree of satisfaction were recorded. Early complications included faecal urgency (3 patients) and pain (2 patients). Complete continence was restored within 10 weeks in all patients except 1 who had persisting incontinence to flatus. All fissures healed completely within 4 weeks. No haemorrhoidal or fissure recurrence has been observed during follow-up. The combination of stapled haemorrhoidopexy and lateral internal sphincterotomy is a safe and effective procedure for the treatment of prolapsing 3rd-degree haemorrhoids and chronic anal fissures. PMID- 16374016 TI - Amino acid changes during successful pregnancy in a case of lysinuric protein insufficiency. AB - Lysinuric protein insufficiency (LPI) is a rare autosomal recessive disorder, and pregnancy in patients with this condition has been considered risky. We, however, observed a relatively favorable course of pregnancy in a woman with LPI, and even amelioration of symptoms during pregnancy. We believe that this unique observation is not only important for patients with LPI who are facing a choice of child-bearing, but is also of interest to obstetricians and researchers of amino acid metabolism. PMID- 16374017 TI - Combining 2nd-trimester maternal serum homocysteine levels and uterine artery Doppler for prediction of preeclampsia and isolated intrauterine growth restriction. AB - AIM: To evaluate the efficacy of a combined 2nd-trimester maternal serum homocysteine and uterine artery Doppler screening at 20 weeks of gestation for complications of pregnancy: preeclampsia, isolated intrauterine growth restriction (IUGR), placental abruption, and stillbirths. METHODS: Consecutive singleton pregnancies without previous risk factors who had homocysteine measured as part of a serum-screening program for trisomy 21 had uterine artery Doppler performed. Sensitivity, specificity, positive and negative predictive values, odds ratio, and positive and negative likelihood ratios for the subsequent development of preeclampsia, isolated IUGR, placental abruption, stillbirth, and preterm delivery were calculated for the following methods (1) homocysteine cut off level 6.3 micromol/l (95th centile); (2) on Doppler ultrasound bilateral notches with a mean resistance index (RI) >0.55 (50th centile), all unilateral notches with a mean RI >0.65 (80th centile), and absence of notches with a mean RI >0.7 (95th centile), and (3) Doppler ultrasound notch evaluation (bilateral, unilateral, absence as in method 2) combined with the homocysteine cut-off level of 6.3 micromol/l. RESULTS: By using a logistic regression model, methods 1 and 2 predicted preeclampsia (p < 0.001), isolated IUGR (p < 0.01), and "any complication" (p < 0.01). The sensitivity for prediction of preeclampsia using the combined method (3) was 61.3% for a false-positive rate of 2%, better than that for isolated IUGR (54%) below the 5th centile and "any complication" (56%). CONCLUSION: This prospective study confirms the potential of a combined method of elevated homocysteine and uterine artery Doppler screening for preeclampsia, isolated IUGR, and any obstetric complication. PMID- 16374018 TI - Postprandial adiponectin levels are unlikely to contribute to the pathogenesis of obesity in Prader-Willi syndrome. AB - AIM: To investigate fasting and postprandial adiponectin levels in PWS patients as compared to obese and lean subjects and whether they could contribute to the pathogenesis of obesity in this syndrome. METHODS: We studied 7 patients with PWS, 16 obese patients and 42 lean subjects for the fasting study. From this group, we evaluated 7 patients with PWS, 7 age-sex-BMI-matched obese non-PWS patients and 7 age-sex-matched lean subjects before and after the administration of 3,139.5 kJ (750 kcal) of a standard liquid meal (53.2% carbohydrate, 30% fat, 16.7% protein) after an overnight fast. Blood samples were obtained every 15 min for the first hour and every 30 min thereafter until 6 h. Adiponectin, IGF-I, glucose, triglycerides, cholesterol, and insulin were measured. RESULTS: Fasting plasma adiponectin levels were lower in PWS than in lean subjects (5.24+/-2.56 vs. 8.28+/-4.63 microg/ml, p=0.041) but higher than in obese patients (4.01+/ 1.27 microg/ml, p=0.047). After the meal, adiponectin concentrations mildly decreased in PWS at time point 240 min, while in obese and lean subjects no changes were observed. However, 6-hour postprandial AUC for adiponectin was similar in all three groups. CONCLUSION: Fasting adiponectin levels are low in PWS, but they are so mildly modulated postprandially that these changes do not seem significant for the pathogenesis of obesity in this syndrome. PMID- 16374019 TI - Age is an important determinant of the growth hormone response to sprint exercise in non-obese young men. AB - BACKGROUND: The factors that regulate the growth hormone (GH) response to physiological stimuli, such as exercise, are not fully understood. The aim of the present study is to determine whether age, body composition, measures of sprint performance or the metabolic response to a sprint are predictors of the GH response to sprint exercise in non-obese young men. METHODS: Twenty-seven healthy, non-obese males aged 18-32 years performed an all-out 30-second sprint on a cycle ergometer. Univariate linear regression analysis was employed to evaluate age-, BMI-, performance- and metabolic-dependent changes from pre exercise to peak GH and integrated GH for 60 min after the sprint. RESULTS: GH was elevated following the sprint (change in GH: 17.0 +/- 14.2 microg l(-1); integrated GH: 662 +/- 582 min microg l(-1)). Performance characteristics, the metabolic response to exercise and BMI were not significant predictors of the GH response to exercise. However, age emerged as a significant predictor of both integrated GH (beta = -0.547, p = 0.003) and change in GH (beta = -0.448, p = 0.019) after the sprint. CONCLUSION: In non-obese young men, age is a more important predictor of GH following sprint exercise than BMI, sprint performance or the metabolic response to sprint exercise. PMID- 16374020 TI - Increased expression of matrix metalloproteinase-2, matrix metalloproteinase-9 and matrix metalloproteinase-13 in lesional skin of bullous pemphigoid. AB - BACKGROUND: Matrix metalloproteinase (MMP)-2, MMP-9 and MMP-13 can degrade type IV collagen which is the major component of the basement membrane zone (BMZ). In bullous pemphigoid (BP), the separation occurs within the BMZ. OBJECTIVE: To evaluate the involvement of MMPs in the pathogenesis of BP, we examined the expression of MMP-2, MMP-9 and MMP-13 in the lesional skin of BP patients. METHODS: The expression of MMP-2, MMP-9, MMP-13, tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 was analyzed by immunohistochemistry in the lesional skin of BP patients in comparison with that in normal human skin. Next, the cellular sources of MMP-2, MMP-9 and MMP-13 were analyzed by double immunohistochemistry. Finally, the levels of these MMPs in the serum and blister fluid of BP patients were measured by ELISA. RESULTS: The number of cells expressing MMP-2, MMP-9 and MMP-13 were significantly increased in the lesional skin of BP patients as compared to that in normal skin. Although the number of cells expressing TIMP-1 and TIMP-2 were also increased in the lesional skin of BP patients as compared to that in normal skin, the ratio of MMPs to TIMPs in the lesional skin of BP patients was high (2.4:1). T cells comprised the major source of MMP-2, MMP-9 and MMP-13, while a proportion of mast cells and eosinophils also expressed these MMPs. Furthermore, marked expression of MMP-2 was detected in the epidermal keratinocytes. The levels of these MMPs in the blister fluid were significantly greater than those in the serum. CONCLUSION: These results suggest that MMP-2, MMP-9 and MMP-13 may be involved in the mechanism of blister formation in BP and that besides infiltrating inflammatory cells, structural cells like epidermal keratinocytes may also participate in the induction of blister formation in BP. PMID- 16374021 TI - Effect of omalizumab on adenosine 5'-monophosphate responsiveness in subjects with allergic asthma. AB - BACKGROUND: The objective of this study was to evaluate the effects of omalizumab on bronchoconstriction induced by methacholine and adenosine 5'-monophosphate (AMP). METHODS: Thirty-four subjects with mild to moderate allergic asthma were randomized to receive placebo (n = 16) or omalizumab (n = 18) subcutaneously during 12 weeks. Airway responsiveness to AMP was measured at baseline and after 4 and 12 weeks of treatment, whereas the response to methacholine was measured at baseline and after 12 weeks of treatment. RESULTS: After 4 weeks of treatment, the increase in AMP PC(20) (provocative concentration required to produce a 20% fall in FEV(1)) was significantly greater in the omalizumab group than in the placebo group, the mean difference in the change between the groups being 1.52 doubling concentrations (95% CI, 0.25-2.79, p = 0.02). Compared with baseline, the mean AMP PC(20) values at 12 weeks were increased by 1.91 doubling concentrations with omalizumab (p < 0.001) and 1.01 doubling concentrations with placebo (p = 0.16), but changes were not significantly different between the treatment groups. Changes in methacholine PC(20) values were not significantly different between the omalizumab and placebo groups. CONCLUSIONS: In subjects with allergic asthma, omalizumab reduces the response to AMP without decreasing the response to methacholine. These findings are consistent with the conclusion that the contribution of IgE to the development of AMP bronchoconstriction is more important than their role in the induction of methacholine hyperresponsiveness. PMID- 16374022 TI - Encasement of bedding does not improve asthma in atopic adult asthmatics. AB - AIM: We evaluated the impact of impermeable bed covers on asthma in asthmatics with clinically relevant house dust mite (HDM) sensitization. METHODS: The study included 32 HDM-sensitized asthmatics in whom HDM allergy was considered as a significant factor in their asthma. They were randomized into either an intervention group whose bedding was encased with impermeable covers, or a control group who received cotton covers. Before and 3 and 6 months after encasement, dust samples were collected from the bedding and assayed for Der p 1. Clinical outcomes included quality of life, lung function, bronchial reactivity to methacholine, symptoms, medications and peak flow rates. RESULTS: Baseline Der p 1 levels in both the active and the placebo groups were comparable and high (19.2 vs 18.9 microg/g of dust). There was a significant reduction in Der p 1 levels in the active group after 6 months, but not in the placebo group (7.3 vs 21.9 microg/g of dust). Quality of life improved significantly in both the intervention and control groups, but there was no significant difference in the improvements between the groups. There was no significant change in lung function, symptoms, and requirements for medications. CONCLUSIONS: Encasement of bedding significantly reduced the Der p 1 levels. However, this was not sufficient to produce worthwhile clinical improvement in those in whom dust mite avoidance might well have been recommended as part of their clinical management. PMID- 16374023 TI - Validity of parent-reported height and weight for defining obesity among asthmatic and nonasthmatic schoolchildren. AB - BACKGROUND: The relationship between parent-reported and measured height and weight is not well established in schoolchildren. This relationship has never been studied in asthmatic children. The objective of this study is to test the validity of the parent-reported weight and height for defining obesity by BMI and to know whether the perception of this height and weight changes when the child suffers from asthma. METHODS: All classes of children of the target ages of 6-8 years (n = 1,672, participation rate 70.2%) of all schools in four municipalities of Murcia (Spain) were included. Parents were asked about their children's weight and height using a questionnaire which included the International Study of Asthma and Allergies in Childhood (ISAAC) core questions on asthma. Parents were not aware that their children were going to be weighed and measured within 1 week's time. Measurements were performed using a rigid stadiometer to the nearest 0.1 cm and a scale to the nearest 0.1 kg. RESULTS: The bias (reported minus real) was, respectively, for nonasthmatics and asthmatics: weight +0.42 kg (95% CI +0.24; +0.59 kg) versus +0.97 kg (+0.50; +1.44 kg), height +2.37 cm (+2.06; +2.68 cm) versus +2.87 cm (+1.87; +3.87 cm); BMI -0.39 kg/m(2) (-0.52; -0.23 kg/m(2)) versus -0.23 kg/m(2) (-0.58; +0.13 kg/m(2)). Diagnostic accuracy of obesity calculated from reported measurements was, respectively, for nonasthmatics and asthmatics: sensitivity 78.0 versus 77.8%, specificity 96.2 versus 94.5%, positive predictive value 77.2 versus 73.7% and negative predictive value 96.4 versus 91.7%. CONCLUSIONS: Reported weights and heights had large biases, comparable between parents of both asthmatic and those of nonasthmatic children. However, this information could be reasonably valid for classifying children as obese or nonobese in large epidemiological studies. PMID- 16374024 TI - Total IgE levels do not change 1 year after endoscopic sinus surgery in patients with chronic rhinosinusitis. AB - BACKGROUND: Total IgE levels positively correlate with the amount of mucosal thickening on sinus CT scans. Our objective was to investigate whether the levels of total serum IgE decreased 1 year after endoscopic sinus surgery in patients with chronic rhinosinusitis, suggesting that the total IgE was influenced by the sinus disease. METHODS: 55 patients about to undergo endoscopic sinus surgery for chronic rhinosinusitis were enrolled in a prospective clinical study. All patients had preoperative sinus computerized tomography (CT) scans and levels of total serum IgE measured before surgery and 1 year postoperatively. RESULTS: Preoperative total IgE levels showed a significant correlation with the extent of disease on sinus CT (r(s) = 0.413, p = 0.002). Total serum IgE levels did not show any statistically significant change from the preoperative values when measured 1 year postoperatively (324.25 +/- 217.30 ng/ml vs. 326.35 +/- 204.50 ng/ml; p = 0.61). CONCLUSIONS: The levels of total serum IgE do not change after sinus surgery for chronic rhinosinusitis. IgE levels in chronic rhinosinusitis may reflect a systemic factor in disease pathogenesis, and are probably not related to the amount of local inflammation within the sinuses. PMID- 16374025 TI - Safety and tolerability of ultra-rush induction, less than one hour, of sublingual immunotherapy in children. AB - BACKGROUND: The safety and tolerability of sublingual immunotherapy (SLIT) has been documented in allergic patients both in the build-up phase as well as during maintenance, but only two studies have evaluated the occurrence of adverse reactions with an ultra-rush regimen of SLIT induction in a mixed paediatric/adult population. Moreover one of these two studies used a chemically modified extract (allergoid). The aim of the present study was to evaluate the occurrence of immediate or late adverse reactions in allergic children after a very fast (40 min) ultra-rush SLIT induction with two different allergen extract solutions. METHODS: We studied 100 children (64 boys, mean age of 9.6 years, range 3.5-16.8), with a history of intermittent/persistent rhinitis and/or intermittent/mild persistent asthma due to inhalant allergens. The ultra-rush build-up phase involved the administration, every 10 min, of increasing doses of the highest-concentration vial of SLIT of two different manufacturers (Anallergo and Stallergenes). RESULTS: All patients completed the treatment, side-effects have been recorded in 19% of the cases: 10% within 1 h after the build-up phase, 7% within 48 h and 2% mixed. A major difference (p = 0.0001) was recorded between Anallergo (6 patients, 8.7%) and Staloral (13 patients, 41.9%), but all the reactions were mild: principally oral symptoms, in 1 case rhinorrhoea and cough, and delayed abdominal pain and diarrhoea in another patient. CONCLUSIONS: No severe adverse reactions were observed with this ultra-rush SLIT induction also in the paediatric age; statistical differences have been documented between the two different extracts. PMID- 16374026 TI - Safety of an ultra-rush immunotherapy build-up schedule with therapeutic vaccines containing depigmented and polymerized allergen extracts. AB - BACKGROUND: Four administration schedules of immunotherapy have been reported: conventional, cluster, rush and ultra-rush. OBJECTIVES: To evaluate the safety and the clinical advantage of using standardized modified allergen extracts in an ultra-rush protocol without premedication and/or hospitalization. MATERIAL AND METHODS: One thousand and sixty-eight patients with rhinoconjunctivitis and/or asthma sensitized to mites and/or pollen were included in a prospective observational study. Patients received a therapeutic vaccine containing depigmented and glutaraldehyde-modified extracts (mites and/or pollens) adsorbed onto alum prescribed by a specialist. The schedule of administration consisted of injecting 0.2 and 0.3 ml of the vial of maximum concentration during the first day of immunotherapy, separated by a time interval of 30 min. All patients reached the maximum dose (0.5 ml) after 2 injections. Tolerance was assessed by recording all side reactions related to immunotherapy, classified according to the criteria of the EAACI. RESULTS: The total number of injections was 2,136. All patients reached the maximum established dose on the 1st day. No premedication was used. Seven clinically relevant local reactions were recorded. The systemic reactions were 5 grade-1 (2 immediate and 3 delayed) and 3 delayed grade-2 reactions. CONCLUSIONS: The therapeutic vaccines containing chemically modified extracts can be administered using an alternative ultra-rush schedule in an immunotherapy unit, reaching the maximum dose on the 1st day with 2 injections, without the need of premedication and/or hospitalization. PMID- 16374027 TI - On the relationship between calculus and caries. PMID- 16374028 TI - The structure, function and properties of the acquired pellicle. PMID- 16374029 TI - Model parameters and their influence on the outcome of in vitro demineralisation and remineralisation studies. PMID- 16374030 TI - Tooth wear. PMID- 16374031 TI - Nanomechanics, chemistry and structure at the enamel surface. PMID- 16374032 TI - Plaque as a reservoir for active ingredients. PMID- 16374033 TI - Vitamins A, E and C as non-enzymatic antioxidants and their relation to lipid peroxidation in children with chronic renal failure. AB - BACKGROUND: Increased lipid peroxidation (LP) and reduced enzymatic antioxidant defense have been observed in predialysis patients with advanced chronic renal failure (CRF) and in patients on maintenance hemodialysis (HD). To extend these observations, we evaluated the plasma, erythrocyte and dialysate levels of vitamins A and E and the plasma and dialysate levels of vitamin C as exogenous non-enzymatic antioxidants in children with CRF treated conservatively and on HD. The data obtained were related to LP monitored by erythrocyte malonyldialdehyde (E-MDA) and plasma organic hydroperoxide (OHP) concentrations. PATIENTS: Forty six predialysis children were enrolled in the study and divided into 2 groups: group I = moderate CRF (plasma creatinine < 265.3 micromol/l), and group II = plasma creatinine > or = 265.3 micromol/l. Group III consisted of 21 HD children. 27 age-matched healthy subjects served as a control group. RESULTS: The plasma levels of vitamin A and vitamin C were significantly reduced in all CRF patients when compared to the controls, with the lowest values observed in children on maintenance HD (group III). Significant differences were also noted between the moderate CRF (group I) and HD (group III). Plasma levels of vitamin E were significantly decreased in moderate CRF (group I) and HD (group III) as compared to controls. In contrast, the erythrocyte vitamin A and vitamin E levels of predialysis children and HD patients were not different from the controls. The E MDA and OHP concentrations in the 3 groups of CRF children were significantly higher than in healthy subjects. The concentration of plasma vitamin C was significantly inversely correlated with E-MDA, plasma OHP and creatinine in group I. In group II we found a significant correlation of plasma vitamin E levels with creatinine and E-MDA and a correlation of the plasma vitamin C concentration with E-MDA. CONCLUSION: CRF in children is associated with decreased concentrations of plasma antioxidant vitamins. This reduction is most expressed in children on maintenance HD and particularly concerns plasma vitamin C and erythrocyte vitamin E concentrations. The low levels of plasma vitamin A, E and C might result in reduced activity of the non-enzymatic antioxidant defense system and might be responsible for increased oxidative stress occurring in children with CRF. PMID- 16374034 TI - Efficacy of interferon-alpha in the treatment of chronic hepatitis C in dialysis patients: two therapeutic protocols compared. AB - BACKGROUND: Data on the efficacy of particular therapeutic protocols of interferon-alpha (IFN-alpha) treatment for chronic hepatitis C in patients on hemodialysis (HD) vary. AIM: To compare the efficacy of two different therapeutic protocols for HD patients. PATIENTS AND METHODS: 15 hepatitis C virus (HCV) positive patients on chronic HD at two dialysis centers: 8 patients treated with IFN-alpha 3 x 3 MU/week s.c. for 6 months (group A), and 7 patients treated with IFN-alpha 3 x 5 MU/week for 3 months, then 1 x 5 MU/week for another 3 months (group B). End of treatment response (ETR) and sustained virologic response (SVR) were evaluated by HCV-RNA determination. There was no statistically significant difference between the two patient groups according to age, sex, duration of HD and HCV infection. RESULTS: ETR was 87.5% (7/8) in group A and 28.5% (2/7) in group B, being statistically significant (p < 0.05). Although better SVR [50% (4/8) vs. 28.5% (2/7)] and lower drop-out rate [0% (0/8) vs. 28.5% (2/7)] were achieved in group A compared to group B, these differences did not reach statistical significance (p > 0.05). CONCLUSION: Therapy with IFN-alpha 3 x 3 MU/week s.c. for 6 months seems to be more appropriate for treatment of hepatitis C in HD patients, mostly due to better tolerability, i.e. lower drop-out rate. These differences could be attributed to different pharmacokinetic properties of the particular therapy protocol. PMID- 16374035 TI - Multiple sclerosis mortality and patterns of comorbidity in the United States from 1990 to 2001. AB - Multiple sclerosis (MS) is a neurodegenerative condition that can result in cognitive and physical disability and shortened life expectancy. However, population-based information is lacking regarding the mortality burden from MS in the United States. We investigated trends in MS mortality rates and examined important comorbidities in the United States from 1990 to 2001. MS deaths were matched by age, sex, and race/ethnicity with randomly selected deaths from other conditions for matched odds ratio comparisons. The overall age-adjusted mortality rate from MS was 1.44/100,000 population. MS mortality rates increased throughout the study period. MS mortality rates were higher in whites than in any other racial/ethnic group, followed by Blacks, Hispanics, American Indians/Alaska Natives, and Asians and Pacific Islanders. Observed mortality rates were more than 10 times lower in Asians and Pacific Islanders than in whites. The odds of pressure ulcers, urinary tract infections, and pneumonia/influenza being reported on the death certificate were higher in MS deaths than in matched controls. PMID- 16374036 TI - Prevalence of sensory and cognitive disabilities and falls, and their relationships: a community-based study. AB - This study assessed the prevalence of sensory and cognitive disabilities and falls for various age groups, sexes, and socio-occupational categories, and their associations in the Lorraine population. The sample included 6,159 subjects, aged 15 years or more, randomly selected from the Lorraine population. They filled in a mailed questionnaire including socio-demographic characteristics, job, falls with physical injuries at the time of the survey, and sensory and cognitive disabilities. Data analysis was made via the adjusted odds ratios. The prevalences of sensory and cognitive disabilities were 3.0 and 4.7%, respectively, that of falls with physical injuries 2.4%: 1.2% for falls on the same level and 1.2% for falls to a lower level. Subjects aged over 70 were markedly affected, but the other age groups had relatively high prevalences as well; the workmen had the highest prevalence, followed by the foremen, farmers, craftsmen and tradesmen, and employees. A twofold risk independently of age, sex and job was found for the subjects with sensory or cognitive disabilities for all falls combined. These findings show the high prevalences of sensory and cognitive disabilities and falls with physical injuries, and their strong associations in the general population. Preventive measures are needed to detect and follow up the persons with these disabilities, limit their risk of falls and reduce occupational hazards. PMID- 16374037 TI - Connective tissue growth factor-mediated angiotensin II-induced hypertrophy of proximal tubular cells. AB - BACKGROUND: Cellular hypertrophy is an early, important pathological feature of renal diseases such as diabetic nephropathy and remnant kidney. Recent studies have demonstrated that angiotensin II (AngII) plays a key role in mediating cell hypertrophy. The aim of our work was to explore the role of connective tissue growth factor (CTGF) in mediating AngII-induced tubular cell hypertrophy in vivoandin vitro. METHODS: In an in vivo study, male Sprague-Dawley rats were randomly divided into three groups: control rats, diabetic rats and diabetic rats treated with irbesartan (IRB). The index of kidney hypertrophy (kidney weight/body weight, KW/BW), glomerular tuft area (AG), glomerular tuft volume (VG) and proximal tubular area (AT) were determined. Renal expression for CTGF was detected by immunohistochemical staining. In an in vitro study, the influence of CTGF antisense oligonucleotide (CTGF AS) on AngII-induced CTGF expression and cell hypertrophy was also investigated. RESULTS: In an in vivo study, diabetic rats showed a significant increase of KW/BW, AG, VG, and AT from week 1 onwards compared to normal control, which could be significantly inhibited by using IRB. Furthermore, there was a significantly increasing expression of CTGF in both glomeruli and tubuli in diabetic rats compared to control, and the extent of CTGF expression closely correlated with the severity of renal hypertrophy. Treatment with IRB could markedly inhibit the renal expression of CTGF. In an in vitro study, AngII stimulated the expression of CTGF mRNA and CTGF protein. AngII significantly increased the total protein content in HK2 cells, which was markedly inhibited by co-treatment with CTGF AS. The average cellular diameter determined by scanning electronic microscope showed that the increase of cell size induced by AngII could be significantly inhibited by CTGF AS. Furthermore, flow cytometer study showed that AngII arrested the cell cycle in the G0-G1 phase, which was significantly reversed by treatment with CTGF AS. CONCLUSION: Our data provide both in vivo and in vitroevidence that CTGF is involved in mediating AngII-induced renal hypertrophy. PMID- 16374038 TI - Expression profiling of crystal-induced injury in human kidney epithelial cells. AB - BACKGROUND: Deposition of crystals within tubular lumens is a feature of many kidney stone diseases, including crystals of calcium oxalate monohydrate (COM) in primary hyperoxaluria and of 2,8-dihydroxyadenine (DHA) in adenine phosphoribosyltransferase deficiency. Crystals are injurious to renal epithelial cells, but the molecular bases of cell injury have not been well characterized. METHODS: We used a cDNA microarray to identify the time-dependent changes in gene expression associated with the interaction of COM or DHA crystals with primary cultures of normal human kidney cortical epithelial cells. RESULTS: We observed gene expression changes that were common to both crystal types, as well as a number of crystal-specific responses. A subset of genes known to be aberrantly expressed in kidney tissue from stone formers also showed an altered expression in COM- or DHA-treated normal human kidney cortical epithelial cells. CONCLUSIONS: Our results show that cultured epithelial cells exposed to COM or DHA crystals demonstrate cellular responses that may be physiologically relevant, thus suggesting that this experimental system may be useful for elucidating the mechanisms of crystal-induced renal cell injury. PMID- 16374039 TI - New guidelines for clinical stage I testicular seminoma? AB - Clinical stage I (CS I) seminoma has been the subject of various studies aimed at finding the ideal treatment. Due to its high radiosensitivity, radiotherapy has been the standard approach for decades. However, the fact that CS I seminoma has a recurrence rate of only 15-20% has prompted many suggestions for better treatment stratification offering surveillance therapy for a subgroup of patients. Moreover, carboplatinum-based monochemotherapy has been the topic of various retrospective studies demonstrating equal effectiveness for adjuvant chemotherapy with one cycle of carboplatin. Since seminoma affects men in their mid-30s, only few reports have been published on long-term survival with reference to radiotoxicity and secondary malignancies. Long-term side effects and secondary malignancies pose an increasing problem for long-term survivors of primary malignancies treated by chemo- or radiotherapy. This becomes evident after a follow-up of 15 years. Since recent studies have disclosed unexpectedly high long-term morbidity and mortality after irradiation, this article critically reviews the value of radiotherapy and carboplatinum-based chemotherapy as equally effective treatment options for CS I seminoma. PMID- 16374040 TI - A dose escalation study of pegylated liposomal Doxorubicin (caelyx) in combination with capecitabine (xeloda) in patients with refractory solid tumors. AB - OBJECTIVE: Pegylated liposomal doxorubicin (PLD) and capecitabine (CAP) have separately shown significant antitumor activity in a wide range of solid tumors. A phase I study was conducted in order to determine the maximum tolerated dose (MTD) and dose-limiting toxicities (DLTs) of their combination in patients with refractory solid tumors. PATIENTS AND METHODS: Fifteen patients with histologically confirmed inoperable solid neoplasms were enrolled. The patients' median age was 65 years, 10 were male, and 12 had a performance status score (WHO) of 0-1. PLD was administered on day 1 as a 1-hour intravenous infusion at escalated doses ranging from 35 to 40 mg/m(2). CAP was administered on days 1-14 per os, at escalated doses ranging from 1,600 to 1,800 mg/m(2), given as two daily divided doses. Treatment was repeated every 3 weeks. RESULTS: At the dose of PLD 40 mg/m(2) and CAP 1,800 mg/m(2), all 3 enrolled patients presented DLTs [2 patients grade 3 palmar-plantar erythrodysesthesia (PPE) and 1 patient grade 3 asthenia] and thus, the recommended MTD for future phase II studies is PLD 40 mg/m(2) and CAP 1,700 mg/m(2). A total of 57 treatment cycles were administered. Grade 2/3 neutropenia complicated 9 (17%) cycles and 1 patient was hospitalized for febrile neutropenia. There was no septic death. The main nonhematologic toxicity was PPE grade 2 in 3 (19%) patients and grade 3 in 4 (27%). PPE was the reason of treatment interruption for 3 patients. Other toxicities were mild and easily manageable. Two patients (16%) with partial response suffering from gastric cancer and 5 patients with (42%) stable disease were observed among 12 evaluable patients. CONCLUSIONS: The results of this phase I study demonstrate that PLD and CAP can be combined at clinically effective and relevant doses. However, PPE is a common side effect and further investigation is warranted to define its precise role in the treatment of solid malignancies. PMID- 16374041 TI - Cortical and subcapsular cataracts: significance of physical forces. AB - Cortical cataracts usually begin with either sharp limited clear fluid clefts, resulting in opaque spokes, or clear lamellar separations, resulting in cuneiform opacities. They do not commence prior to 45 years of age. From this age on an increase in lens nuclei hardening can be detected. Therefore, during disaccommodation in older lenses, mechanical shear stresses must develop between the soft remaining cortices and the harder nuclei. These shear stresses are significant regarding the different cortical ruptures in predisposed lenses: in a privileged radial direction (according to zonular traction) of the sharp limited cortical spokes, or in parallel microridges at the commencement of lamellar separations, as observed when a rubber surface slides against a harder object. In pure cortical cataracts the ion pump (K+ > Na+) and investigated metabolic parameters remain largely intact. Therefore, it is not surprising that, in contrast to subcapsular cataracts, subcapsular opacities do not occur. Subcapsular cataracts are known to be caused by a variety of factors: aging, diabetes, corticosteroids, iridocyclitis, or X-ray, among many others. In contrast to cortical cataracts, subcapsular cataracts were found to be closely associated with ion pump damage (Na+ > K+) and a variety of metabolic activity alterations. In subcapsular cataracts passive fluids (from the vitreous and camera anterior) enter externally through the lens capsule. This initially forms numerous free clear, secondary grey, subcapsular fluid vacuoles. If the ion pump (metabolic barrier) is more severely damaged fluids may also enter the lens nucleus (secondary grey nuclear cataract), which rarely results in intumescent cataract. In cortical and subcapsular cataracts and lens perforations the main cause of grey opalescence appears to be the result of lens proteins (water soluble crystalline) coming into direct contact with free fluids (water). In cortical cataracts this happens in the area of sharp limited mechanical cortical ruptures (fluid clefts), and in subcapsular cataracts during passive, external fluid entry, resulting in subcapsular fluid vacuoles and opacities, and also later grey-white nuclear opacities. The importance of water contact with water soluble lens crystallines in behalf of light scattering and turbidness also has been investigated experimentally. PMID- 16374042 TI - Experimental posterior perforating ocular injury in rabbit eyes: effects of radiotherapy with or without intravitreal cyclosporin on vitreous proliferation. AB - Double perforating ocular injuries were made in 30 rabbits and the effects of radiotherapy (RT) alone or in combination with cyclosporin (CS) on vitreous proliferation investigated. Thirteen rabbits in group 1 received RT alone (600 cGy), and 13 rabbits in group 2 received RT combined with 100 microg of intravitreal CS. No treatment was given to 4 rabbits in the control group (group 3). All animals were ophthalmologically examined on the 1st, 3rd, 7th, 14th and 28th days following the injury and the clinical findings scored; they were sacrificed on the 28th day, and histopathological scoring was made. The median histopathological score of the RT group (4.0; range: 0-8) was significantly higher than that of the CS + RT group (1.0; range: 0-5; p = 0.018). We conclude that intravitreal CS may be used as an adjunctive agent to RT to inhibit intraocular proliferation following penetrating ocular injury in rabbits. PMID- 16374043 TI - Serum paraoxonase 1 activity and lipid peroxidation levels in patients with age related macular degeneration. AB - Our objective was to investigate antioxidant paraoxonase 1 (PON1) activity together with malondialdehyde (MDA) levels to evaluate oxidative stress in patients with age-related macular degeneration (AMD), an important cause of blindness in the elderly population. Serum PON1 activity and MDA levels were analyzed in 37 patients with AMD and compared with 29 healthy controls using a spectrophotometric method. Serum MDA levels were significantly higher in the patient group (2.76 +/- 1.28 nmol/ml) than controls (1.00 +/- 0.36 nmol/ml; p < 0.001), whereas PON1 activity was lower in the patient group (132.27 +/- 63.39 U/l) than controls (312.13 +/- 136.23 U/l; p < 0.001). There was a negative correlation between MDA and PON1 levels (r = -0.470, p < 0.001). We conclude that the observed increase in MDA levels may be related to decreased PON1 activity; the present data also demonstrated that an obvious negative correlation between PON1 activity and MDA levels exists in patients with AMD. PON1 is also an antioxidant agent, therefore effective antioxidant therapy to inhibit lipid peroxidation is necessary and agents to increase PON1 activity may be a therapeutic option in AMD. PMID- 16374044 TI - Inhibition of experimental proliferative vitreoretinopathy with protein kinase C inhibitor (chelerythrine chloride) and melatonin. AB - PURPOSE: To investigate whether a protein kinase C (PKC) inhibitor and melatonin prevent proliferative vitreoretinopathy (PVR). METHODS: Twenty pigmented rabbits were used in this study. All rabbits except controls received an intravitreal injection of 0.15 ml (75,000 units) of platelet-rich plasma into their left eye. The animals were divided into four groups: group I was treated with intravitreal injection of 0.1 ml (100 micromol/ml) of PKC inhibitor (chelerythrine chloride), group II received 1 ml (4 mg/kg) of intraperitoneal melatonin for 3 days, group III received nothing (blank group), and group IV (control group) received only 0.5 ml of 1% ethanol intraperitoneally for 3 days. Proliferative changes were graded in a masked fashion by indirect ophthalmoscopy for a 15-day follow-up period. The malondialdehyde (MDA), reduced glutathione (GSH) and total nitrite levels were measured in the vitreous humor. RESULTS: The grades of PVR were A and B in group I and II, treated with PKC inhibitor and melatonin, respectively. The PVR grade in the blank group was C-D. The mean MDA level in group I (4.2 +/- 0.9 micromol/l) was significantly lower than in the blank group (6.0 +/- 1.0 micromol/l; p < 0.05). The mean GSH level in group I (66.3 +/- 8.8 micromol/l) was not significantly different from that in the blank group (p > 0.05). The MDA and GSH levels in group II were 3.2 +/- 0.7 and 70.1 +/- 13.3 micromol/l, respectively. Both these levels were significantly different from those of the blank group (p < 0.05). The NO levels in both treatment groups were significantly lower than in the blank group (p < 0.001). CONCLUSION: These findings suggest an inhibitory effect of PKC inhibitor and melatonin on PVR. The inhibition of PVR development was associated with lower MDA and NO levels with higher GSH levels in the treatment groups. PMID- 16374045 TI - Confirmation of the adult-onset primary open angle glaucoma locus GLC1B at 2cen q13 in an Australian family. AB - Primary open-angle glaucoma (POAG) is genetically heterogeneous, with 6 named POAG loci GLC1A-F mapped and genes myocilin (MYOC) and optineurin (OPTN) identified at 2 of the loci. Using penetrance-model-free methods, we screened the POAG loci GLC1A-F in an extended Australian pedigree, using 3-5 markers within each locus. p values of less than 0.05 were obtained empirically using SimWalk2 and exactly using Genehunter for 2 markers within the GLC1B region on chromosome 2. Fine mapping of this region produced p values of 0.01 or less at 5 markers flanked by D2S1897 and D2S2269. The 9 cM haplotype of interest overlaps the original GLC1B region. These results provide supportive evidence for the GLC1B locus on chromosome 2cen-q13 and verify the existence of POAG susceptibility gene in this region, increasing the likelihood of gene identification. PMID- 16374046 TI - Retinal angiomatous proliferation: association with clinical and angiographic features. AB - AIM: It was the aim of this study to evaluate the frequency of retinal angiomatous proliferation (RAP) and its association with specific clinical and angiographic characteristics in age-related macular degeneration (AMD). METHODS: A consecutive series of 126 newly diagnosed patients with exudative AMD was reviewed retrospectively. All underwent a complete ophthalmic examination, a red free photograph and fluorescein angiography. Most patients (85/126) underwent indocyanine green choroidal angiography (ICGA). RAP was diagnosed when a connection between the retinal vasculature and the neovascular complex was recognized angiographically. RESULTS: Out of 126 patients with recent neovascular AMD, 17 had RAP (13.5%; 95% CI 8.1-20.7). The study eye of patients with RAP had more frequent hemorrhages (88.2 vs. 59.6%; p = 0.027), hard exudates (82.4 vs. 26.6%; p < 0.001), pigment epithelium detachment (64.7 vs. 23.8%; p = 0.001) and a hot spot in ICGA (70.6 vs. 22.1%; p < 0.001) with respect to the other forms of exudative AMD. Hemorrhages were more frequently superficial, multiple and within the edge of the lesion in the RAP group. Bilateral AMD was more common in the RAP group (70.6 vs. 38.0%; p = 0.011). No statistically significant differences were found regarding sex, age and visual acuity. CONCLUSION: RAP represents a common lesion in patients with neovascular AMD referred to a tertiary care clinic. The recognition of hemorrhages, hard exudates, pigment epithelium detachment or a hot spot in ICGA can assist a correct diagnosis. PMID- 16374047 TI - Comparison of forward shift of posterior corneal surface after operation between LASIK and LASEK. AB - PURPOSE: To compare the forward shift of the posterior corneal surface as a function of time after surgery between successful laser in situ keratomileusis (LASIK) and laser-assisted subepithelial keratectomy (LASEK). METHODS: Twenty-two eyes from 14 patients who underwent LASIK and 19 eyes from 10 patients who underwent LASEK were reviewed retrospectively. The uncorrected visual acuity, spherical equivalent and pachymetry were obtained before and 1 or 2 weeks and 2, 6, 12, and 24 months after surgery. The change in the elevation of the posterior corneal surface was assessed using the difference map of an Orbscan II (Bausch and Lomb, Salt Lake City, Utah, USA), which was generated from the preoperative and subsequent postoperative elevation map. RESULTS: One or 2 weeks after LASIK and LASEK, the posterior corneal surface had shifted forward by 18.00 and 25.90 microm, respectively (p = 0.008). However, the mean posterior corneal surface reduced by 3.05 microm from 1 or 2 weeks to 2 years in the LASIK group (p = 0.359). In contrast, the mean posterior corneal surface had reduced by 12.40 microm in the LASEK group (p = 0.004). This forward shift did not return to the corneal normal elevation observed prior to surgery in the two groups. CONCLUSIONS: Refractive surgery induced a forward shift of the posterior corneal surface early postoperatively in the LASIK and LASEK groups, and this change after LASEK was significantly larger than that observed after LASIK. The posterior corneal surface gradually shifted backwards depending on the time course in both groups, but they did not return to the preoperative levels. PMID- 16374048 TI - Clinical analysis of benign eyelid and conjunctival tumors. AB - PURPOSE: To assess the relative frequency and clinicopathologic characteristics of benign conjunctival tumors. METHODS: A retrospective study of 80 consecutive patients admitted to our hospital with benign eyelid and conjunctival tumor between April 2000 and November 2002 was undertaken, and clinical records including age, sex and involved site of tumors and pathology slides of the patients were reviewed retrospectively. RESULTS: Twenty-three males and 33 females presented with benign eyelid tumors and 12 males and 12 females with conjunctival tumors. Mean age was 42.3 and 29.7 years, respectively. The lower eyelid was involved in 27 (48.2%) of eyelid tumors and the medial conjunctiva in 14 (58.3%) of conjunctival tumors. The most frequent tumor was intradermal nevus (44.6%), seborrheic keratosis (16.1%) and compound nevus (10.7%) in eyelid tumors, and compound nevus (29.2%) and intradermal nevus (25.0%) in conjunctival tumors. CONCLUSION: This report will provides a basic analysis of benign eyelid and conjunctival tumors. PMID- 16374049 TI - Oxidative damage to lens in culture: reversibility by pyruvate and ethyl pyruvate. AB - It is generally believed that prophylactic intake of antioxidants is beneficial in delaying the onset of some aging manifestations such as cataract. However, whether such a supplementation will also be effective if the pathophysiological process has already set in remains a largely open question. We examined this possibility with lens changes leading to cataract formation, since cataract genesis is intimately related to a continued generation of reactive oxygen species (ROS) in the aqueous humor. Since the formation of cataract is a well defined progressive disease, starting with an early refractive change and leading to gradual enhancement of opacification, we hypothesized that even a belated start with an appropriate anti-oxidant could halt the pathological process and delay cataract maturation and vision impairment. Using lens cultures, we tested this hypothesis with pyruvate, known to be an effective and highly potent ROS scavenger. Adding pyruvate to the culture medium after lenses had sustained a 50% damage was significantly effective in preventing progress. This was apparent by better maintenance of the active rubidium transport activity in these lenses compared to controls without pyruvate treatment. Glutathione levels were also higher in the pyruvate group. PMID- 16374050 TI - Keratectasia after laser in situ keratomileusis. Clinicopathological case report. AB - PURPOSE: To describe the morphological features of a prominent ectasia of the cornea after laser in situ keratomileusis (LASIK). METHODS: The morphology of the ectatic corneas was examined using corneal topography, light microscopy and transmission electron microscopy in 2 cases who underwent penetrating keratoplasty due to poor visual acuity induced by progressive corneal ectasia after LASIK. RESULTS: On topographic examination, the apex of the corneal surface was observed within the central 3-mm zone, and the smallest thickness was 0.116 and 0.271 mm in each case. On histological examination, the epithelial layer became thinner and detached easily. Bowman's membrane was broken down and folded. An irregular arrangement of the stromal lamellae with fibroblastic keratocytes was found. The fulled fiber cell, a transformed epithelial cell, was visible in a plane on Bowman's layer in the central region. In contrast, the corneal endothelium was intact, and no abnormality was found in both cases. CONCLUSION: On morphological examination of 2 cases with corneal ectasia, a forward protrusion of both the anterior and posterior corneal surfaces occurred, and epithelial detachment, Bowman's membrane breakage and folding and irregular lamellae were found. The 2 cases had greatly thinned and protruding corneas, yet there was no abnormality in the corneal endothelium. PMID- 16374051 TI - Delayed trigeminocardiac reflex induced by an intraorbital foreign body. Case report. AB - OBJECTIVE: To emphasize the importance of the mechanism and surgical approach to trigeminocardiac reflex (TCR) developing 48 h after orbital trauma due to a foreign body. CASE REPORT: After gunshot injury of a 17-year-old male patient, computerized tomography evaluation revealed a right globe perforation and an intraorbital metallic foreign body in the right orbita adjacent to the lateral wall. The ocular perforation was repaired, but the foreign body was not removed. Constant bradycardia (45/min) developed 48 h after the operation. Since there were no cardiological findings, a temporary cardiac pacemaker was inserted and on the 6th postoperative day, the foreign body was removed through orbitolateral approach. After the removal of the foreign body, bradycardia completely recovered. CONCLUSION: In the presence of an intraorbital foreign body accompanied by globe perforation, TCR may develop 48 h after the trauma and insertion of a temporary pacemaker may be required to control the cardiac rhythm. In this paper, the delayed TCR complication presented an indication for the removal of the intraorbital foreign body. PMID- 16374052 TI - Maintaining the redox-balance intact: gosha-jinki-gan but not insulin activates retinal soluble guanylate cyclase in diabetic rats. AB - Strategies to prevent hyperglycemia-induced cytotoxic reactive oxygen species in the retina include the prevention of free radical production, activation of radical-scavenging capacities and inhibition of aldose reductase. This study examined the effect of the standardized Japanese herbal extract product gosha jinki-gan (GJG) in comparison to insulin treatment in the rat retina. Diabetes was induced in male Wistar rats by single injection of streptozotocin (50 mg/kg i.p.). At 6 and 12 weeks, eye-cups were removed for immunohistochemistry. At 12 weeks, lipid peroxidation (tested with the antiacrolein antibody, Ab5F6) was enhanced significantly in the untreated diabetic group. This effect was absent in both treatment groups, notably in the outer retina. A similar result was obtained for nitrotyrosine overproduction. As an early treatment effect, GJG -- but not insulin -- enhanced soluble guanylate cyclase (sGC) activation (using the function-sensing antibody, MoAb 3221). GJG not only reduces nitroxidative stress and lipid peroxidation in the retina, it also ameliorates glucose metabolism within the cells. We propose that the high glucose turnover in the insulin treated model disturbs the intracellular redox equilibrium, one result of which might be the impaired sGC activation. PMID- 16374053 TI - Effect of UVA and UVB irradiation on the metabolic profile of rabbit cornea and lens analysed by HR-MAS 1H NMR spectroscopy. AB - PURPOSE: The aim of the study was to investigate the metabolic profiles of intact rabbit corneas and lenses exposed to UVA and UVB radiation by using high resolution (HR) magic angle spinning (MAS) (1)H nuclear magnetic resonance (NMR) spectroscopy and pattern recognition methods. METHODS: Adult albino rabbits were exposed to UVA (366 nm, 0.589 J/cm(2)) or UVB (312 nm, 1.667 J/cm(2)) radiation for 8 min, once a day for 5 days. Three days after the last irradiation day, samples of corneas and lenses were dissected. HR-MAS (1)H NMR spectroscopy combined with pattern recognition methods (principal component analysis and soft independent modelling of class analogy) and one-way ANOVA were applied to obtain metabolic information from intact corneal and lens tissue. RESULTS: UVB irradiation caused statistically significant metabolic changes in the rabbit corneas. A decrease in metabolites as ascorbate (84%), myo-inositol (59%), hypotaurine (91%) and choline (76%) was observed. Exposure to UVA radiation caused no significant metabolic alteration in this tissue. The metabolic profile of the rabbit lenses showed no detectable changes after UVA or UVB exposure. CONCLUSIONS: The combination of HR-MAS (1)H NMR spectroscopy and multivariate methods proved effective to analyse intact corneal and lens tissue after exposure to UV radiation of different wavelengths. By avoiding extraction methods and obtaining complete metabolic profiles from one sample, HR-MAS (1)H NMR spectroscopy provided important information about metabolic alteration occurring in rabbit corneal and lens tissue after UV exposure. PMID- 16374054 TI - rd1 mouse retina shows imbalance in cellular distribution and levels of TIMP 1/MMP-9, TIMP-2/MMP-2 and sulfated glycosaminoglycans. AB - BACKGROUND: The rd1 mouse retina displays fast degeneration of photoreceptors resulting in a depletion of almost all rod photoreceptors by postnatal day 21 (PN21). To evaluate the role of proteinases in the pathophysiology of this animal model of retinitis pigmentosa, C3H rd1 and congenic wild-type (wt) mice retinas were analyzed. MATERIAL AND METHODS: The cellular localization and levels of proteins, matrix metalloproteinases (MMPs), their endogenous inhibitors (TIMPs), total sulfated glycosaminoglycans (sGAG) and nature of saccharides in rd1 and wt retinal extracts were compared. RESULTS: MMP-2/TIMP-2 and MMP-9/TIMP-1 were predominantly localized in the interphotoreceptor matrix (IPM) of both genotypes, but MMP-2/TIMP-2 also appeared in the Muller cell fibers of rd1 retina. In rd1 retinal extracts the levels of total proteins were lower and those of active MMP 9, MMP-2, TIMP-1 and total sGAG were higher than those of wt extracts. Despite an increase in TIMP-1, active MMP-9/MMP-2 were disproportionately elevated in rd1 compared to wt retina. With increasing age, MMPs in wt retinas were decreased but were increased in rd1. The sialylation of proteoglycans in PN2 and PN7 rd1 retinas was lower, and galactosylation was higher than that in wt retinas. CONCLUSIONS: MMP-9/MMP-2 and TIMP-1/TIMP-2 are associated with IPM, possibly after secretion by retinal pigmented epithelial cells. In degenerating rd1 retina, MMP-2/TIMP-2 are associated with the Muller cell fibers, which apparently play a central role in modifying the balance between MMPs and TIMPs. Elevated sGAG and proteolysis due to an imbalance in the levels of TIMPs and active MMP 9/MMP-2 in rd1 retina possibly contribute to retinal degeneration in the rd1 mouse. PMID- 16374056 TI - Expression of cochlin in the vestibular organ of rats. AB - The COCH gene mutated in autosomal dominant sensorineural deafness (DFNA9) encodes cochlin, a major constituent of the inner ear extracellular matrix. Cochlin constitutes 70% of the inner ear protein and cochlin isoforms can be classified into three subgroups, p63s, p44s and p40s. Symptoms of some DFNA9 patients are consistent with those of Meniere's disease. Here, we report the expression of cochlin in the vestibular organ of rats using isoform-specific antibodies that recognize all three isoforms. Cochlin is highly expressed in the stromata of the maculae of otolithic organs and cristae of semicircular canals, and in the channels in the bony labyrinth that transmit the dendritic innervation to the cristae and maculae. Cochlin cannot be detected in the sensory cells, dark cells, nor in the acellular structures, otolithic membrane or in the cupula. These findings support the theory that deposition of acidophilic substance in the inner ear caused by mutation of cochlin can induce a secondary retrograde dendritic degeneration of the vestibular nerves. PMID- 16374057 TI - CGRP expression in the vestibular periphery after transient blockage of bilateral vestibular input. AB - This study aimed to establish an animal model of reversible bilateral vestibular disorders that is suitable for examining the mechanisms of vestibular plasticity, and to observe the changes in the plasticity of vestibular efferent systems. Tetrodotoxin (TTX) was infused continuously for 7 days into the bilateral perilymph of guinea pig cochlea. We assessed the vestibulo-ocular reflex (VOR) for evaluating the vestibular function. We also investigated the changes in calcitonin gene-related peptide (CGRP) immunoreactivity in vestibular end organs to observe the changes in the plasticity of vestibular systems. The VOR was completely eliminated by TTX administration and returned to the preoperative levels within 120 h after TTX discontinuation. An obvious increase in the number of CGRP-immunoreactive fibers was observed within the neurosensory epithelia of the maculae and cristae. An animal model of reversible bilateral vestibular disorders was established and used for investigating the plasticity of the vestibular nervous system. PMID- 16374058 TI - Contribution of endolymphatic fluid shift to caloric response in plugged semicircular canals. AB - The purpose of this study was to clarify the role of endolymphatic fluid shift in caloric response, using frog posterior semicircular canals (PSCs). PSCs were sutured using 10-0 nylon thread and were used as a model of canal plugging. Compound action potentials (CAPs) of the PSC nerve evoked by a cooling stimulus were recorded. The CAPs after suturing the PSCs were found to be greater than those before suturing. This indicates that the fluid shift effect increases after canal suturing. Additionally, we present a clinical case in which caloric nystagmus was observed after lateral canal plugging. In this case MRI revealed the fluid space from the plugged portion toward the ampulla to be intact. There was another case with lateral canal plugging that showed the same findings on MRI. The above findings support the hypothesis that fluid shift is responsible for the caloric response without the convective flow of endolymph in the plugged canal. PMID- 16374059 TI - A new method for drug application to the inner ear. AB - Inner ear sensory cells are very susceptible to injuries and recovery after damage is very difficult. Recently several drugs including neurotrophic factors have been reported to protect against inner ear injury. The purpose of this experimental study is to find new methods for applying drugs to the inner ear that effectively protect against inner ear damage. Biodegradable hydrogel was used as a carrier for application of brain-derived neurotrophic factor (BDNF) into the inner ear of guinea pigs through the round window membrane. After application of BDNF the number of surviving spiral ganglion neurons increased following injury of inner ear hair cells and spiral ganglion neurons by ototoxic treatment. This result indicates that BDNF provides effective protection against inner ear damage and that biodegradable hydrogel is useful for application of drugs to the inner ear. PMID- 16374060 TI - Isosorbide delays gentamicin-induced vestibular sensory cell death. AB - The efficacy of isosorbide for protection from vestibular sensory cell damage was investigated. The effects of isosorbide on gentamicin-induced production of nitric oxide (NO) and reactive oxygen species (ROS) were studied by means of the fluorescence indicators 4,5-diaminofluorescein diacetate and dihydrotetramethylrosamine. The effect on gentamicin-induced vestibular sensory cell damage was examined by using an in vitro LIVE/DEAD system. Isosorbide inhibited the production of both NO and ROS. Isosorbide limited the vestibular sensory cell damage caused by gentamicin. It is, therefore, suggested that isosorbide may help to treat inner ear disorders. PMID- 16374061 TI - Effects of lithium on endolymph homeostasis and experimentally induced endolymphatic hydrops. AB - There is evidence to suggest that water homeostasis in the inner ear is regulated via the vasopressin (VP)-aquaporin 2 (AQP2) system in the same fashion as in the kidney. The VP-AQP2 system in the kidney is well known to be inhibited by lithium, resulting in polyuria due to a decrease in reabsorption of water in the collecting duct of the kidney. Therefore, lithium is also likely to inhibit the VP-AQP2 system in the inner ear, and consequently exert some influence on inner ear fluid homeostasis. In this study, we investigated the effects of lithium on AQP2 expression in the rat inner ear, and on the cochlear fluid volume in hydropic ears of guinea pigs. A quantitative PCR study revealed that lithium reduced AQP2 mRNA expression in the cochlea and endolymphatic sac. Lithium application also decreased the immunoreactivity of AQP2 in the cochlea and endolymphatic sac. In a morphological study, lithium intake significantly reduced endolymphatic hydrops dose-dependently. These results indicate that lithium acts on the VP-AQP2 system in the inner ear, consequently producing a dehydratic effect on the endolymphatic compartment. PMID- 16374062 TI - Meniere's disease is associated with single nucleotide polymorphisms in the human potassium channel genes, KCNE1 and KCNE3. AB - Although the bases for both the sporadic and inherited forms of Meniere's disease (MD) remain undefined, it is likely to be multifactorial, one of the factors being a genetic predisposition. Recently, genetic association studies on complex diseases have become very popular and most of them are case-control studies using single nucleotide polymorphisms (SNPs) as markers. Mutations/polymorphisms in KCNE potassium channel genes might play a causative role in MD, because KCNE potassium channels have been suggested to be present and active in transmembrane ion and water transports in the inner ear. In the present study, to identify MD susceptibility genes, we have conducted a genetic association study with optimized sampling, optimized phenotyping/genotyping, and a selection of KCNE genes as the candidate genes. The SNPs analyses identified 112G/A SNP in the KCNE1 gene and 198T/C SNP in the KCNE3 gene in 63 definite MD cases as well as 205 and 237 non-MD control subjects. For both KCNE1 and KCNE3 genes, a significant difference in frequency of each SNP was confirmed between MD cases and non-MD control subjects. The result indicates that 112G/A SNP in the KCNE1 gene and 198T/C SNP in the KCNE3 gene could determine an increased susceptibility to develop MD. PMID- 16374063 TI - Microarray analysis of stress-related gene expression in patients with Meniere's disease. AB - We developed an original microarray carrying 1,467 cDNAs of stress-related genes for the assessment of stress responses. In this study, we used microarray analysis to assess the stress-related gene expression profiles in peripheral leukocytes in 2 patients with definite Meniere's disease. In the attack and active phases, mRNA expression levels of 57 genes and 163 genes were either up regulated more than twofold or down-regulated by less than half in patient 1 and patient 2, respectively. Patient 1 had sporadic episodes of vertigo attack, while patient 2 had an intractable course with frequent vertigo attacks, suggesting that the magnitude of changes in gene expression is correlated with the severity of the disorder in Meniere's disease. The expression of a total of 26 genes commonly changed in both patients in the attack and active phases and returned to the baseline levels in the remission phase, suggesting the involvement of the distinct group of stress-related genes in the development of vertigo attacks in Meniere's disease. We then examined the effects of caloric stimulation on the stress-related gene expression profiles in peripheral leukocytes in 5 healthy volunteers. Although unilateral caloric stimulation with cold water caused acute vertigo with nystagmus, the expression profiles of stress-related genes did not significantly change after this experiment. This finding indicated that the up- or down-regulated genes in the attack and active phases in patients with Meniere's disease are not secondary to vertigo or vertigo-associated anxiety. All these findings suggested that the distinct group of stress-related genes contributed to the development of vertigo attacks of Meniere's disease and that stress-related gene expression profiles in peripheral leukocytes can be a predictive and therapeutic tool for episodic vertigo attacks in patients with Meniere's disease. PMID- 16374064 TI - Personal factors involved in onset or progression of Meniere's disease and low tone sensorineural hearing loss. AB - PURPOSE: To identify personal causative factors for Meniere's disease. PROCEDURES: Patterns of hearing loss progression were studied in patients with Meniere's disease and low-tone sensorineural hearing loss, and the involvement of stress and the relation of stressors to the onset or progression of the disease were analyzed. RESULTS: Low-tone loss recurred in 40% of patients even after hearing was restored, and low-tone loss progressed to high-tone loss after frequent repetitions of recovery and recurrence. High-tone loss tended to proceed to all-tone loss. Eighty percent of patients reported that stress was involved or deeply involved in the onset or progression of the disease. Common causative stressors were business-related pressure, insufficient sleep, and troubles at the workplace or at home. CONCLUSIONS: The present findings indicate that recovery and recurrence may be influenced by the strength and duration of stress that is produced when patients do not feel rewarded for engrossment in their work or for self-inhibiting behaviors. PMID- 16374065 TI - Epidemiologic characteristics of definite Meniere's disease in Japan. A long-term survey of Toyama and Niigata prefectures. AB - To identify epidemiologic characteristics of definite cases of Meniere's disease (DMD), we conducted retrospective surveys of the period 1990-2004 of the Nishikubiki district and of the period 1980-2004 of Toyama Medical and Pharmaceutical University. Three hundred and seventy-five patients (50 from Nishikubiki, 325 from Toyama) were diagnosed with DMD according to the diagnostic criteria proposed by the Japanese Society for Equilibrium Research. There was a slight increase in the prevalence of DMD during the period 1990-2004. However, incidence did not change significantly over time. The average annual prevalence and incidence were 34.5 and 5.0, respectively, per 100,000 population. Incidence and prevalence predominated in females. With respect to age at disease, the incidence in elderly patients was increased when we corrected for age distribution in the overall population. PMID- 16374066 TI - Paraffin-embedded cell line microarray (PECLIMA): development and validation of a high-throughput method for antigen profiling of cell lines. AB - The introduction of high-throughput techniques is increasingly providing abundant information on molecular alterations requiring validation at the posttranscriptional level. Protein expression is now efficiently evaluated in large series of tumors included in tissue microarrays. We propose, describe and validate a technique to elaborate paraffin-embedded cell line microarrays (PECLIMA) from fixed cell cultures, which can be processed like standard surgical pathology biopsies prior to immunophenotyping. Our results show a reliable protein immunoexpression profiling in six widely used cell lines under different fixation conditions. This technique permits the simultaneous analysis of multiple antigens in multiple cell lines under different experimental conditions. Additional features of these arrays are long-term storage, their suitability for a variety of techniques including immunocytochemistry and in situ hybridization and their low cost. PMID- 16374067 TI - Expression of liver-type fatty-acid-binding protein, fatty acid synthase and vascular endothelial growth factor in human lung carcinoma. AB - OBJECTIVE: A key enzyme of fatty acid synthesis, fatty acid synthase (FAS), is expressed in human cancers, including squamous-cell carcinoma of the lung, and long-chain fatty acids are intracellularly transported and/or taken up from blood by fatty-acid-binding proteins (FABPs). Since the liver-type (L-) FABP, a member of the FABPs, is detected in a subset of gastric adenocarcinomas, the expression of FAS, L-FABP and vascular endothelial growth factor (VEGF) was investigated in human lung carcinomas to elucidate the mechanisms of production and transportation of fatty acid(s) in cancer. METHODS: Expression of L-FABP, FAS and VEGF in 199 surgically resected lung carcinomas was examined immunohistochemically. Possible associations of the expression of each protein with major clinicopathological factors were analyzed. RESULTS: L-FABP was detected in 60% (120 of 199) of the lung carcinoma cases; detection was increased in large-cell carcinoma (80%) and adenosquamous carcinoma (83%), but low in squamous-cell carcinoma (47%) and in small-cell carcinoma (57%). Overall expression of FAS was 67.3% (134 of 199 cases) and that of VEGF was 86.8% (158 of 199 cases), respectively. Expression of L-FABP was not correlated with the FAS status, but there was a tendency to co-expression of L-FABP and VEGF. There was no association between L-FABP, FAS or VEGF expression and clinicopathological data. CONCLUSIONS: L-FABP, FAS and VEGF are highly expressed in human lung cancer, and expression of L-FABP is associated with that of VEGF but not that of FAS, suggesting that L-FABP might be involved in the uptake of fatty acid(s) from the bloodstream. PMID- 16374068 TI - Overexpression of cyclooxygenase-2 is associated with a favorable prognostic phenotype in breast carcinoma. AB - OBJECTIVES: Cyclooxygenase (COX) is the rate-limiting enzyme that catalyzes the conversion of arachidonic acid to prostaglandins. The purpose of the present study was to investigate the involvement of COX-2 protein in breast cancer biological behavior through its correlation with the well-known clinicopathological parameters and the expression of p53, c-erbB-2, topoisomerase IIalpha (topoIIalpha) and peroxisome proliferator-activated receptor (PPARgamma) proteins, as well as its effect on patients' survival. METHODS: We performed immunohistochemistry to detect COX-2, estrogen receptor (ER), progesterone receptor (PR), p53, c-erbB-2, topoIIalpha and PPARgamma proteins in 175 cases of invasive breast carcinomas. The results were elaborated by statistic analysis. RESULTS: Cytoplasmic expression of COX-2 was detected in 66.9% of breast carcinoma samples and was inversely correlated with both nuclear and histological grade (p < 0.0001 and p = 0.039, respectively), whereas its association with PR was found to be positive (p = 0.016). COX-2 expression was inversely correlated with topoIIalpha and p53 (p = 0.033 and p = 0.002, respectively), whereas its association with PPARgamma was parallel (p < 0.0001). In addition, c-erbB-2 of tumor cells was inversely correlated with COX-2 in stromal cells of the tumor (p = 0.011). Neither univariate nor multivariate analysis demonstrated any association between COX-2 expression and patient overall or disease-free survival. CONCLUSIONS: The current data suggest that increased expression of COX 2 may be related to breast carcinomas with less aggressive phenotype. This suggestion is further supported by the positive correlation between COX-2 and PPARgamma, since the latter is considered to be indicative of a less malignant phenotype of tumor cells. PMID- 16374069 TI - Basement membrane matrix modifies cytokine interactions between lung cancer cells and fibroblasts. AB - OBJECTIVE: Proliferation of fibroblasts (desmoplastic reaction) in the lung adenocarcinomas is an important phenomenon that correlates with metastases and poor prognosis. Because basement membranes are often involved in the desmoplastic areas and many cytokines have binding capacity to basement membrane molecules, we hypothesized that basement membrane modify the paracrine effects between cancer cells and fibroblasts via the fibrogenic cytokines and this hypothesis was experimentally investigated. METHODS: The effects of conditioned media derived from ten lung carcinoma cell lines and normal airway epithelial cells on DNA synthesis of fetal lung fibroblasts were determined. We focused on fibroblast growth factor 2 (FGF-2) as the candidate paracrine cytokines and examined their diffusion through an experimental basement membrane matrix model, Matrigel. RESULTS: All the conditioned media promoted DNA synthesis of fetal lung fibroblasts. Detection by ELISA methods and the neutralizing antibodies suggested that FGF-2 was one of the responsible factors for the growth promotion. Diffusion of FGF-2 across the polycarbonate membrane was suppressed by coating with Matrigel. When FGF-2-secreting A549 cells were covered with Matrigel, FGF-2 was stored in Matrigel and its diffusion into the culture media was significantly reduced. Binding of FGF-2 to Matrigel was completely blocked by a basic protein, protamine sulfate. In the presence of protamine sulfate in Matrigel overlaid on A549 cells, diffusion of FGF-2 increased 7-fold as much as that without overlaid Matrigel. CONCLUSION: These results suggest that the basement membrane acts as a barrier to the diffusion and a reservoir of cytokines secreted by cancer cells, and that the subsequent degradation of the basement membrane by cancer cells could release the stored cytokines and promote growth of fibroblasts. PMID- 16374070 TI - Fas ligand expression and its correlation with apoptosis and proliferation in Lobund-Wistar prostate carcinomas. AB - OBJECTIVE: The Fas (CD95) interaction with its receptor Fas ligand (FasL) is one of the main mechanisms of cell apoptosis. High expression of FasL has been consistently observed in a variety of human cancers. In this study, we evaluated FasL and its relationship with apoptosis and proliferation in Lobund-Wistar (L-W) cancers. The L-W rat strain develops spontaneous and induced adenocarcinomas in the anterior prostate and seminal vesicles. Although FasL expression has been observed in a subset of human prostate carcinomas, this multistage model allowed in vivo evaluation of subclones of malignant cells with a single genetic susceptibility. METHODS: Apoptosis was evaluated in spontaneous, induced and transplanted tumors as well as metastasis using the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) technique and transmission electron microscopy. Proliferating cell nuclear antigen (PCNA) and FasL expression were detected using immunohistochemistry and analyzed according to the number of positive cells and intensity of staining using a semiquantitive method. RESULTS: Apoptotic indexes were significantly higher in spontaneous tumors compared to induced (p < 0.008), transplanted tumors (p < 0.0112) and metastases (p < 0.009). TUNEL-positive cells were frequently observed in the leukocytic infiltrate of the stroma in transplanted carcinomas and metastases. These findings were confirmed by electron microscopy. FasL expression was not uniformly localized in L-W carcinomas and its highest expression was observed in transplanted tumors and metastasis (p < 0.005). Moreover, PCNA indices were directly correlated with cancers showing high FasL total scores (Hscores). CONCLUSIONS: In this model, high FasL expression was associated with cells displaying low apoptotic indexes and high PCNA index. Therefore, analysis of FasL may have clinical relevance in detecting the malignant potential of prostate cancers. PMID- 16374071 TI - Immunohistochemical analysis of morules in colonic neoplasms: morules are morphologically and qualitatively different from squamous metaplasia. AB - Morules develop in several neoplasms and have been considered as a type of squamous metaplasia despite the absence of keratinization and intercellular bridges. The objective of this study was to clarify the pathological significance of morules and to distinguish morules from squamous metaplasia in colonic neoplasms. Ten cases of morule-associated colonic neoplasms (4 adenocarcinomas, 1 adenoma with carcinoma in situ, and 5 adenomas), and 3 cases of squamous metaplasia in colonic adenocarcinoma were examined morphologically and immunohistochemically. Morules were well-defined structures composed of small, oval to short-spindled cells with bland nuclei, and frequently associated with intranuclear inclusions that were positive for biotin and biotin-binding enzymes (pyruvic acid carboxylase and propionyl CoA carboxylase). On immunohistochemical examination, morules characteristically showed nuclear overexpression of beta catenin, cyclin D1 and p63, low Ki-67 labeling index (<1%), cytoplasmic overexpression of CD10, and no expression of cytokeratin 20. These molecules were useful for the differentiation of morules. Furthermore, p63 and 34betaE12 positivities in morules suggested that they have a basal/stem cell phenotype. Thus, morules were morphologically and qualitatively different from squamous metaplasia. We consider that morules in colonic neoplasms are cell clusters with a basal/stem cell phenotype, and have less proliferative and less invasive potential than other cancer cells. PMID- 16374072 TI - An investigation into the presence of a vagal tachycardia and the effect of vasoactive intestinal polypeptide on rat heart rate in vitro. AB - The presence of the vagal tachycardia and the effect of vasoactive intestinal polypeptide in the isolated innervated rat atrium were investigated. The right vagus, or cardiac branch, were stimulated at 4, 8, 16 and 32 Hz, pulse duration 1 ms, 20 V, 30 s before atropine and for 1 min after atropine (3 micromol/l), experiments were carried out in the presence of atenolol (4 micromol/l). No significant vagal tachycardia was observed in the presence of atropine, the greatest increase in heart rate was at 16 Hz which was 3+/-1 beats/min (n = 12 rats) (p = 0.052). Baseline heart rates for the control, 226+/-11 beats/min (n = 12 rats) and atropine experiments, 210+/-8 beats/min (n = 12 rats), were not significantly different (p = 0.24). VIP (0.06, 0.12, 0.24 micromol/l) caused a maximum increase of 27+/-13 beats/min (n = 5 rats) after 6 micromol/l VIP which was not significant, two higher concentrations of VIP failed to increase heart rate further. These results show that the vagal tachycardia is not present and that VIP does not cause a significant tachycardia in the rat. PMID- 16374073 TI - Hemodynamic effects of di-sec-butyl phenol, an anesthetic substituted phenol. AB - Dose- and age-related hemodynamic effects were determined for an anesthetic substituted phenol, 2,6-di-sec-butyl phenol (DSB). DSB, 7.5 mg/kg, induced hypnosis in young rabbits and increased mean blood pressure to 170 +/- 14% and heart rate to 150 +/- 21% of control values. In elderly rabbits, 7.5 mg/kg DSB induced hypnosis, had no effect on blood pressure, but increased the heart rate to 130 +/- 2% of control. After ganglionic blockade with hexamethonium, 7.5 mg/kg DSB caused a decline in mean blood pressure (71 +/- 5% of control) without change in heart rate. DSB increased norepinephrine release from SH-SY5Y cells, a human neuroblastoma cell line (5.4 +/- 1.7% vs. 3.5 +/- 0.3%). DSB produced age dependent elevation of mean blood pressure in rabbits, probably by causing release of catecholamines from the sympathetic nervous system. PMID- 16374074 TI - Deep brain stimulation inactivity can produce unexpected high electrode impedances when reactivated, leading to a false conclusion of wire fracture. AB - Two months after bilateral DBS surgery in a patient with PD, the stimulator had to be removed temporarily due to an infection. After successful antibiotic treatment, a new stimulator and extension wire were reimplanted 2 months later. Intraoperative testing of the replacement device yielded unusually high electrode impedances and low currents on one lead across all four contacts. Since the results did not change on repeated measurements, a lead fracture was suspected. Visual inspection of the exposed hardware, however, did not reveal any obvious defects. It was then decided to activate the lead for a prolonged time with significant currents. Subsequent impedance measurements yielded normal values and after the patient awoke from anesthesia, good symptom control was again achieved. It is advisable if a lead fracture is suspected under similar circumstances to 'condition' the electrode with high currents. Failure to do so could lead to a false conclusion of a lead fracture and unnecessary stereotactic surgery to replace it. PMID- 16374076 TI - Soft copy digital mammography. PMID- 16374077 TI - Simple pulmonary eosinophilia evaluated by means of FDG PET: the findings of 14 cases. AB - OBJECTIVE: We wanted to describe the findings of simple pulmonary eosinophilia with using 18 fluorodeoxyglucose (FDG) positron emission tomography (PET). MATERIALS AND METHODS: We analysed the findings of 14 patients who underwent thoracic computed tomography (CT) and PET, and then they were subsequently proven to have simple pulmonary eosinophilia. PET studies were performed in four patients with malignancy to evaluate for cancer metastasis, and PET scans were also done in 10 healthy subjects who underwent volunteer cancer screening. The PET scans were evaluated by using the maximum standardized uptake values (SUVs). The subjects' CT findings also were reviewed and correlated with the PET findings. RESULTS: A total of 42 nodules were detected on the CT scans. There were single nodules in three patients and multiple nodules in 11 patients (mean number of nodules: 3, range: 1-10, mean diameter: 9.5 mm+/-4.7). Twelve of 42 (28.6%) nodules showed FDG uptake and their mean maximum SUV was 2.5+/-1.6 (range: 0.6-5.3). Five of six solid nodules showed FDG uptake (2.2+/-1.1, range: 0.9-3.6), six of 11 semisolid nodules showed FDG uptake (3.1+/-1.8, range: 0.6 5.3) and one of 25 pure ground-glass opacity nodule showed a maximum SUV of 0.8. The maximum SUVs of seven nodules in five patients were greater than 2.5. The maximum SUVs were significantly different according to the nodule types (p < 0.001). CONCLUSION: Simple pulmonary eosinophilia commonly causes an increase in FDG uptake. Therefore, correlation of the PET findings with the CT findings or the peripheral eosinophil counts can help physicians arrive at the correct diagnosis of simple pulmonary eosinophilia. PMID- 16374078 TI - Screen-film mammography and soft-copy full-field digital mammography: comparison in the patients with microcalcifications. AB - OBJECTIVE: We wanted to compare the ability of screen-film mammography (SFM) and soft-copy full-field digital mammography (s-FFDM) on two different monitors to detect and characterize microcalcifications. MATERIALS AND METHODS: The images of 40 patients with microcalcifications(three patients had malignant lesion and 37 patients had benign lesion), who underwent both SFM and FFDM at an interval of less than six months, were independently evaluated by three readers. Three reading sessions were undertaken for SFM and for FFDM on a mammography-dedicated review workstation (RWS, 2K x 2.5K), and for FFDM on a high-resolution PACS monitor (1.7K x 2.3K). The image quality, breast composition and the number and conspicuity of the microcalcifications were evaluated using a three-point rating method, and the mammographic assessment was classified into 4 categories (normal, benign, low concern and moderate to great concern). RESULTS: The image quality, the number and conspicuity of the microcalcifications by s-FFDM (on the RWS, PACS and both) were superior to those by SFM in 85.0%, 80.0% and 52.5% of the cases, respectively (p < 0.01), and those by the s-FFDM on the two different monitors were similar in 15.0%, 12.5% and 35.0% of the cases, respectively (p > 0.01). The mammographic assessment category for the microcalcifications in the three reading sessions was similar. CONCLUSION: s-FFDM gives a superior image quality to SFM and it is better at evaluating microcalcifications. In addition, s-FFDM with the PACS monitor is comparable to s-FFDM with the RWS for evaluating microcalcifications. PMID- 16374079 TI - Mammographic mass detection using a mass template. AB - OBJECTIVE: The purpose of this study was to develop a new method for automated mass detection in digital mammographic images using templates. MATERIALS AND METHODS: Masses were detected using a two steps process. First, the pixels in the mammogram images were scanned in 8 directions, and regions of interest (ROI) were identified using various thresholds. Then, a mass template was used to categorize the ROI as true masses or non-masses based on their morphologies. Each pixel of a ROI was scanned with a mass template to determine whether there was a shape (part of a ROI) similar to the mass in the template. The similarity was controlled using two thresholds. If a shape was detected, then the coordinates of the shape were recorded as part of a true mass. To test the system's efficiency, we applied this process to 52 mammogram images from the Mammographic Image Analysis Society (MIAS) database. RESULTS: Three hundred and thirty-two ROI were identified using the ROI specification methods. These ROI were classified using three templates whose diameters were 10, 20 and 30 pixels. The results of this experiment showed that using the templates with these diameters achieved sensitivities of 93%, 90% and 81% with 1.3, 0.7 and 0.33 false positives per image respectively. CONCLUSION: These results indicate that the detection performance of this template based algorithm is satisfactory, and may improve the performance of computer-aided analysis of mammographic images and early diagnosis of mammographic masses. PMID- 16374080 TI - Gadobenate dimeglumine as an intrabiliary contrast agent: comparison with mangafodipir trisodium with respect to non-dilated biliary tree depiction. AB - OBJECTIVE: To compare the efficacy of Mangafodipir trisodium (Mn-DPDP)-enhanced MR cholangiogrphy (MRC) and Gadobenate dimeglumine (Gd-BOPTA)-enhanced MRC in visualizing a non-dilated biliary system. MATERIALS AND METHODS: Eighty-eight healthy liver donor candidates underwent contrast-enhanced T1-weighted MRC. Mn DPDP and Gd-BOPTA was used in 36 and 52 patients, respectively. Two radiologists reviewed the MR images and rated the visualization of the common duct, the right and left hepatic ducts, and the second-order branches using a 4-point scale. The contrast-to-noise ratio (CNR) of the common duct to the liver in the two groups was also compared. RESULTS: Mn-DPDP MRC and Gd-BOPTA MRC both showed similar visualization grades in the common duct (p=.380, Mann-Whitney U test). In the case of the proximal bile ducts, the median visualization grade was significantly higher with Gd-BOPTA MRC than with Mn-DPDP MRC (right hepatic duct: p=0.016, left hepatic duct: p=0.014, right secondary order branches: p=0.006, left secondary order branches, p=0.003). The common duct-to-liver CNR of the Gd-BOPTA MRC group was significantly higher (38.90+/-24.50) than that of the Mn-DPDP MRC group (24.14+/-17.98) (p=.003, Student's t test). CONCLUSION: Gd-BOPTA, as a biliary contrast agent, is a potential substitute for Mn-DPDP. PMID- 16374081 TI - Percutaneous treatment of extrahepatic bile duct stones assisted by balloon sphincteroplasty and occlusion balloon. AB - OBJECTIVE: To describe the technical feasibility and usefulness of extrahepatic biliary stone removal by balloon sphincteroplasty and occlusion balloon pushing. MATERIALS AND METHODS: Fifteen patients with extrahepatic bile duct stones were included in this study. Endoscopic stone removal was not successful in 13 patients, and two patients refused the procedure due to endoscopy phobia. At first, all patients underwent percutaneous transhepatic biliary drainage (PTBD). A few days later, through the PTBD route, balloon assisted dilatation for common bile duct (CBD) sphincter was performed, and then the stones were pushed into the duodenum using an 11.5 mm occlusion balloon. Success rate, reason for failure, and complications associated with the procedure were evaluated. RESULTS: Eight patients had one stone, five patients had two stones, and two patients had more than five stones. The procedure was successful in 13 patients (13/15). In 12 of the patients, all stones were removed in the first trial. In one patient, residual stones were discovered on follow-up cholangiography, and were subsequently removed in the second trial. Technical failure occurred in two patients. Both of these patients had severely dilated CBD and multiple stones with various sizes. Ten patients complained of pain in the right upper quadrant and epigastrium of the abdomen immediately following the procedure, but there were no significant procedure-related complications such as bleeding or pancreatitis. CONCLUSION: Percutaneous extrahepatic biliary stone removal by balloon sphincteroplasty and subsequent stone pushing with occlusion balloon is an effective, safe, and technically feasible procedure which can be used as an alternative method in patients when endoscopic extrahepatic biliary stone removal was not successful. PMID- 16374082 TI - Efficacy of a dexamethasone-eluting nitinol stent on the inhibition of pseudo intimal hyperplasia in a transjugular intrahepatic portosystemic shunt: an experimental study in a swine model. AB - OBJECTIVE: We wanted to evaluate the feasibility and efficacy of using a dexamethasone (DM)-eluting nitinol stent to inhibit the pseudo-intimal hyperplasia following stent placement in the transjugular intrahepatic portosystemic shunt tract (TIPS) of a swine. MATERIALS AND METHODS: Fifteen stents were constructed using 0.15 mm-thick nitinol wire; they were 60 mm in length and 10 mm in diameter. The metallic stents were then classified into three types; type 1 and 2 was coated with the mixture of 12% and 20%, respectively, of DM solution and polyurethane (PU), while type 3 was a bare stent that was used for control study. In fifteen swine, each type of stent was implanted in the TIPS tract of 5 swine, and each animal was sacrificed 2 weeks after TIPS creation. The proliferation of the pseudo-intima was evaluated both on follow-up portogram and pathologic examination. RESULTS: One TIPS case, using the type 1 stent, and two TIPS cases, using the type 2 stent, maintained their luminal patency while the others were all occluded. On the histopathologic analysis, the mean of the maximum pseudo-intimal hyperplasia was expressed as the percentage of the stent radius that was patent, and these values were 51.2%, 50% and 76% for the type 1, 2, and 3 stents, respectively. CONCLUSION: The DM-eluting stent showed a tendency to reduce the development of pseudo-intimal hyperplasia in the TIPS tract of a swine model with induced-portal hypertension. PMID- 16374083 TI - Prospective comparison of collagen plug (Angio-Seal) and suture-mediated (the Closer S) closure devices at femoral access sites. AB - OBJECTIVE: Rapid and effective hemostasis at femoral puncture sites minimizes both the hospital stay and patient discomfort. Therefore, a variety of arterial closure devices have been developed to facilitate the closure of femoral arteriotomy. The objective of this prospective study was to compare the efficacy of two different closure devices; a collagen plug device (Angio-Seal) and a suture-mediated closure device (the Closer S). MATERIALS AND METHODS: From March 28, 2003 to August 31, 2004, we conducted a prospective study in which 1,676 cases of 1,180 patients were treated with two different types of closure device. Angio-Seal was used in 961 cases and the Closer S in 715 cases. The efficacy of the closure devices was assessed, as well as complications occurring at the puncture sites. RESULTS: Successful immediate hemostasis was achieved in 95.2% of the cases treated with Angio-Seal, and in 89.5% of the cases treated with the Closer S (p < 0.05). The rates of minor and major complications occurring between the two groups were not significantly different. In the Closer S group, we observed four major complications (0.6%), that consisted of one massive retroperitoneal hemorrhage (surgically explored) and three pseudoaneurysms. In the Angio-Seal group, we observed three major complications (0.3%) that consisted of one femoral artery occlusion, one case of infection treated with intravenous antibiotics and one pseudoaneurysm. CONCLUSION: The use of Angio-Seal was found to be more effective than that of the Closer S with regard to the immediate hemostasis of the femoral puncture sites. However, we detected no significant differences in the rate at which complications occurred. PMID- 16374084 TI - Stent placement for chronic iliac arterial occlusive disease: the results of 10 years experience in a single institution. AB - OBJECTIVE: We wanted to retrospectively evaluate the long-term therapeutic results of iliac arterial stent placement that was done in a single institution for 10 years. MATERIALS AND METHODS: From May 1994 to April 2004, 206 patients who underwent iliac arterial stent placement (mean age; 64+/-8.8) were followed up for evaluating the long term stent patency. Combined or subsequent bypass surgery was performed in 72 patients. The follow up period ranged from one month to 120 months (mean; 31+/-25.2 months). The factors that were analyzed for their effect on the patency of stents were age, the stent type and diameter, the lesion site, lesion shape, lesion length, the Society of Cardiovascular and Interventional Radiology criteria, the total run off scores, the Fontaine stage and the cardiovascular risk factors (diabetes mellitus, hypertension and smoking). Follow-up included angiography and/or CT angiography, color Doppler sonography and clinical evaluation with the ankle-brachial index. RESULTS: Two hundred and eighty-four stents were placed in 249 limbs of 203 patients. The technical success rate was 98% (203/206). The primary patency rates of the stents at 3, 5, 7 and 10 year were 87%, 83%, 61% and 49%, respectively. One hundred seventy-seven patients maintained the primary stent patency until the final follow up and 26 patients showed stenosis or obstruction during the follow up. Secondary intervention was performed in thirteen patients. Lesions in the external iliac artery (EIA) or lesions in both the common iliac artery (CIA) and EIA were a poor prognostic factor for stent patency. The run off score and stent diameter also showed statistically significant influence on stent patency. The overall complication rate was 6%. CONCLUSION: Iliac arterial stent placement is a safe treatment with favorable long term patency. Lesions in the EIA or lesions in both the EIA and CIA, poor run off vessels and a stent having the same or a larger diameter than 10 mm were the poor prognostic factors for long term stent patency. PMID- 16374085 TI - Ultrasonography in the diagnosis of appendicitis: evaluation by meta-analysis. AB - OBJECTIVE: We wanted to review the usefulness of ultrasonography (US) for the diagnosis of appendicitis and to evaluate the diagnostic accuracy of US according to patients' and researchers' characteristics. MATERIALS AND METHODS: The relevant Korean articles published between 1985 and 2003 were included in this study if the patients had clinical symptoms of acute appendicitis. The histopathologic findings were the reference standard and the data were presented for 2 x 2 tables. Articles were excluded if patients had no sonographic signs of appendicitis according to graded-compression US. Two reviewers independently extracted the data on study characteristics. The Hasselblad method was used to obtain the combined estimates of sensitivity and specificity for the performance of US. RESULTS: Twenty-two articles (2,643 patients) fulfilled all inclusion criteria. The estimate of d calculated by combining the sensitivity and specificity was 2.0054 (95% confidence interval [CI]: 1.8553, 2.1554) by a random effects model. The overall sensitivity and specificity (95% CI) were 86.7% (85.4 to 88.0), and 90.0% (88.9 to 91.2), respectively. According to the subgroup meta analysis by patients' characteristics, the d estimate (95% CI) of dominantly younger age, male, and highly clinical suggestive group for US was 2.2388 (1.8758 to 2.6019), 2.7131 (2.2493 to 3.1770), and 2.4582 (1.7387 to 3.1777), respectively. Also, according to subgroup meta-analysis by researchers' characteristics, the d value (95% CI) for US done by diagnostic radiologists and gray-scale was 2.0195 (1.7942 to 2.2447) and 2.2630 (1.8444 to 2.6815). CONCLUSION: This evidence suggests that US may be useful for the diagnosis of acute appendicitis, especially when patients are younger age, male, and highly clinical suggestive. PMID- 16374086 TI - Diagnostic importance of 3D CT images in Klippel-Feil Syndrome with multiple skeletal anomalies: a case report. AB - We present here the case of a 12-year-old boy who had Klippel-Feil syndrome with renal, cardiac and multiple skeletal anomalies, and we show the relevent three dimensional computed tomography images. Our patient had a triple renal pelvis, mitral valve prolapsus, multiple cervical vertebrae fusions, cervical ribs, hypoplasia of the right thumb, spina bifida of L5, lumbalization at the right side of S1 and a sacral curved defect. In this study, we discuss the atypical clinical features and the diagnostic value of three-dimensional CT for evaluating the skeletal anomalies of the Klippel-Feil syndrome cases. PMID- 16374087 TI - 2005 Jurkiewicz Lecture. PMID- 16374088 TI - Impact of significant weight loss on outcome of body-contouring surgery. AB - Morbid obesity in the United States has reached startling proportions, with serious physical and psychosocial ramifications. As the number of patients undergoing bariatric surgery increases to treat this problem, it is expected there will be an increase in the number of patients presenting to plastic surgery offices for body contouring after weight loss. It is imperative that plastic surgeons prepare patients with extreme weight loss as to the risks and complications as compared with the general population undergoing body contouring. In this retrospective chart review, 26 patients undergoing body contouring from May 2001 to November 2004 were reviewed. Only patients who had extreme weight loss, defined as losing greater than 50 pounds, were included in this study. The age, gender, amount of weight lost, comorbidities, tobacco use, and weight-loss method were recorded. Additionally, the type of body-contouring procedure and presence of complications were collected from the records. The percentage of complications in our patient population is significantly higher than reported in the literature for the general population undergoing an abdominoplasty. We believe the increased wound complications seen in our patients can be attributed to the inherent complications seen with obese patients. PMID- 16374089 TI - Do progressive tension sutures really decrease complications in abdominoplasty? AB - The purpose of this study is to evaluate the efficacy of progressive tension sutures (PTS) in preventing or reducing seroma formation and local wound complications in patients undergoing abdominoplasty. Forty-nine patients who underwent abdominoplasty procedures with the use of PTS were retrospectively compared with a historical control group of 54 patients who underwent standard abdominoplasty. Primary outcomes measured were seroma formation and local wound complications, including hypertrophic scar formation, umbilical distortion, and wound necrosis. Secondary outcomes were all other complications and time to drain removal. Overall complication rates and local wound complication rates were significantly reduced with the addition of PTS to abdominoplasty. Seroma formation and the time to drain removal were reduced in the PTS group, but the findings were not statistically significant. PMID- 16374090 TI - Implant breast reconstruction using acellular dermal matrix. AB - Thirteen breast reconstructions in 11 patients, averaging 58 years of age, underwent mastectomies. The technique uses a saline implant either totally or partially covered with a human acellular dermal matrix. The mean postoperative follow-up time was 14 months. Ninety percent of the patients were considered high risk; the thickness of the human acellular dermal matrix was an average of 1.3 mm, with an average area per breast of 121 cm. There were 12 successful breast reconstructions (92%) that provided stability, increased soft tissue padding, which allowed a greater resemblance to normal breast shape and decreased rippling and implant visibility. The graft was used in an onlay fashion or as an extension of the pectoralis major muscle that covers the implant. A representative histologic cross-section of well-integrated human acellular dermal matrix is presented. The use of a human acellular dermal matrix in breast reconstruction is an alternative protocol in high-risk patients, resulting in a minimal increase in operative time and a decrease in morbidity compared with more extensive procedures. PMID- 16374091 TI - Breast reduction as an alternative treatment option for early breast cancer in women with macromastia. AB - Macromastia has been considered a relative contraindication to breast conservation therapy because of difficulties with postoperative radiation therapy and cosmesis. This study evaluates the feasibility of the inferior pedicle reduction mammaplasty as a component of breast conservation therapy for patients with early breast cancer. A retrospective review identified 6 patients with macromastia receiving oncologic treatment of breast cancer and simultaneous breast reduction. Mean age was 43.5 +/- 8.7 (mean +/- SD) years, and all breast cancers were stage I or II, averaging 2.3 +/- 1.5 cm in size. All patients underwent a Wise-pattern inferior pedicle breast reduction after cancer extirpation and received postoperative radiation as part of their treatment. They were evaluated for postoperative complications, esthetic outcome of the breasts, and local recurrence. Patients in this series were followed for an average of 30.3 months, with no significant postoperative complications and recurrences. Breast reduction incisions healed primarily and adjuvant radiation was completed without a delay. All patients were pleased with the esthetic result and had improvement of their symptoms related to macromastia. Thus, we believe that breast reduction is a reasonable and safe option for early breast cancer patients with macromastia who desire breast conservation therapy. Our combined oncologic and reconstructive approach may improve the outcome of this group of patients with early breast cancers. PMID- 16374092 TI - Latissimus dorsi flap remains an excellent choice for breast reconstruction. AB - Latissimus dorsi flap has been unfairly relegated to a second option in breast reconstruction. One hundred consecutive latissimus dorsi muscle flaps (LDMF) with tissue-expander reconstruction were studied, mean follow-up 34.5 months (range, 1 175), 50 immediate, 50 delayed. With attention to a few technical details, excellent esthetic, soft reconstructions were achieved. Complications included 1 partial flap loss; 2 patients required inframammary fold revision; and 6 patients required surgery for capsular contracture. Donor-site seroma occurred in 34 patients; 6 required operative revision. Results were similar in the immediate versus the delayed groups. LDMF remains an esthetic, reliable, safe reconstructive choice. PMID- 16374093 TI - Latissimus dorsi breast reconstruction utilizing functional muscle transfer and tissue expansion. AB - Latissimus dorsi breast reconstruction with implants has been criticized for Baker III or IV capsular contractures in as many as 30% of patients. Also criticized is muscle atrophy and loss of breast volume and definition. This study evaluated 2 modifications of the classic latissimus reconstruction: muscle transferred as an innervated functional unit and tissue expansion posterior to the latissimus but anterior to the pectoralis muscle. After expansion, a permanent prosthesis was placed (46 silicone gel and 3 saline). Forty-nine such reconstructions were done in 32 patients (17 bilateral and 15 unilateral), with average follow-up of 19 months. Only 4 Baker III capsular contractures (8%) and no Baker IV contractures occurred. All patients demonstrated excellent retention of breast shape/definition. This study demonstrates that functional muscle transfer and tissue expansion results in low capsular contracture rates and excellent retained breast shape. PMID- 16374094 TI - High septal osteotomy in rhinoplasty for the deviated nose. AB - When attempting to straighten a patient's healed, deviated bony nasal dorsum, deviation of the central structure (high dorsal septum and medial nasal bones) must be addressed following the completion of medial and lateral osteotomies. When hump resection is not performed, blunt fracture (digitally or with forceps) of the deviated central structure is not a reliable method of mobilization, often leading to postoperative nasal drift. An intranasal osteotomy technique to mobilize the central structure of the nose is described, called "high septal osteotomy." Review of 25 cases suggests high septal osteotomy, supplemented as needed by resection of overlapping septal elements, can be performed safely and efficaciously, permitting stable midline reduction of the nasal pyramid. The technique is not advocated when hump resection is performed, as it is unnecessary and could destabilize the dorsum. Even aggressive maneuvers to mobilize the bony dorsum may fail if not performed properly with meticulous attention to completion of all osteotomies. PMID- 16374095 TI - A simple and inexpensive method of preoperative computer imaging for rhinoplasty. AB - GOALS/PURPOSE: Despite concerns of legal liability, preoperative computer imaging has become a popular tool for the plastic surgeon. The ability to project possible surgical outcomes can facilitate communication between the patient and surgeon. It can be an effective tool in the education and training of residents. Unfortunately, these imaging programs are expensive and have a steep learning curve. The purpose of this paper is to present a relatively inexpensive method of preoperative computer imaging with a reasonable learning curve. MATERIALS AND METHODS: The price of currently available imaging programs was acquired through an online search, and inquiries were made to the software distributors. Their prices were compared to Adobe PhotoShop, which has special filters called "liquify" and "photocopy." It was used in the preoperative computer planning of 2 patients who presented for rhinoplasty at our institution. Projected images were created based on harmonious discussions between the patient and physician. Importantly, these images were presented to the patient as potential results, with no guarantees as to actual outcomes. RESULTS: Adobe PhotoShop can be purchased for 900-5800 dollars less than the leading computer imaging software for cosmetic rhinoplasty. Effective projected images were created using the "liquify" and "photocopy" filters in PhotoShop. Both patients had surgical planning and operations based on these images. They were satisfied with the results. CONCLUSIONS: Preoperative computer imaging can be a very effective tool for the plastic surgeon by providing improved physician-patient communication, increased patient confidence, and enhanced surgical planning. Adobe PhotoShop is a relatively inexpensive program that can provide these benefits using only 1 or 2 features. PMID- 16374096 TI - Palatal fistula repair using acellular dermal matrix: the University of Florida experience. AB - Palatal fistulas represent a challenging problem for surgeons caring for patients with cleft palate. The purpose of this study was to examine the rate of fistula closure using conventional surgical techniques (Group 1) versus a newer technique (Group 2) employing the use of acellular dermal matrix (AlloDerm). We reviewed the charts of all patients who underwent palatal fistula repair between July 1994 and February 2005. The surgical techniques in Group 1 varied and were considered a historical control. In the second group, a piece of dermal matrix was interposed between the nasal and oral mucoperiosteum after closure of these layers. The primary fistula closure rate for Group 1 was 83.3% (10/12 patients). Fistula closure was obtained in 100% of the patients in Group 2. There were no complications noted other that the failure of the 2 patients in Group 1. The use of AlloDerm in palatal fistula repair has reduced our failure rate from 16.7% to 0%. The use of AlloDerm during palatal fistula repair is safe, effective over time, and has the potential to reduce palatal fistula recurrence rates. PMID- 16374097 TI - The use of a hydrogel sealant on flexor tendon repairs to prevent adhesion formation. AB - The prevention of peritendinous adhesions after zone II flexor tendon repair poses a significant challenge to hand surgeons. This study evaluates a hydrogel sealant (FocalSeal-L) as a barrier to peritendinous adhesion formation. The deep flexors of toes 2 through 4 were divided and repaired in 30 chickens. Chickens were randomized to tendon repair with (n = 15) or without (n = 15) FocalSeal-L. Each group was further randomized to have their tendons studied postoperatively at 3 (n = 10), 6 (n = 10), or 12 (n = 10) weeks. Histologic evaluation revealed decreased peritendinous adhesion formation in the FocalSeal-L group. Biomechanical analysis demonstrated a decrease in work of flexion in the FocalSeal-L group that was most pronounced at 6 weeks (P = 0.0020). There was no significant difference in breaking strength. Apparently, an effective barrier to peritendinous adhesion formation, this sealant system is easy to use, biocompatible, and bioresorbable. In addition, it is not bulky or restrictive to tendon glide. PMID- 16374098 TI - The reversed medial hemisoleus muscle flap and its role in reconstruction of an open tibial wound in the lower third of the leg. AB - The usefulness of a reversed hemisoleus muscle flap as a local reconstructive option for soft-tissue coverage of an open tibial wound in the lower third of the leg has never been acknowledged. Over the past 2 years, 8 patients underwent soft tissue reconstruction of an open tibial wound (3 x 3 to 10 x 6 cm) in the lower third of the leg with the reversed medial hemisoleus muscle flap modified by the author. The flap was dissected with attention to preserve several critical perforators from the posterior tibial vessels to the flap as possible while allowing adequate turnover of the flap to cover the exposed tibia or hardware. There was no total flap loss, and limb salvage was achieved in all patients. Only 2 patients developed insignificant distal flap necrosis, and they were treated subsequently with debridement and flap readvancement. All patients had reliable healing of their tibial wounds, with good reconstructive and cosmetic outcomes of their flap reconstructions during follow-up. Therefore, the author believes that the reversed medial hemisoleus muscle flap can be a good choice for soft-tissue coverage of a sizable open tibial wound in the lower third of the leg and may be used successfully to replace free tissue transfer in selected patients. PMID- 16374099 TI - Restoration of fecal continence after functional gluteoplasty: long-term results, technical refinements, and donor-site morbidity. AB - PURPOSE: For patients with severe fecal incontinence, reconstruction of the anal sphincter, via gluteoplasty, may improve quality of life, but little is known about long-term functional results. We present our comprehensive experience with gluteoplasty, highlighting technical refinements, donor-site morbidity, and functional outcomes. METHODS: We performed a retrospective analysis of 25 consecutive patients (22 female, 3 male; mean age 42 years, range 23-65) undergoing gluteoplasty for fecal incontinence at a university teaching hospital from 1996-2004. Etiology of incontinence was as follows: obstetrical injury (n = 13), irritable bowel syndrome (n = 3), previous rectal surgery (n = 3), Crohn disease (n = 3), impalement (n = 1), rectocele (n = 1), and idiopathic (n = 1). RESULTS: Gluteoplasty was successful in restoring fecal continence in 18 patients (72%) and was partially successful in 4 patients (16%). Two patients required permanent ostomy because of refractory incontinence. Donor-site morbidity and perirectal complications were observed in 16 patients (64%) and included dysthesias (n = 7), cellulitis (n = 5), irregular contour (n = 3), abscess (n = 2), seroma (n = 2), fistula (n = 1), but no hip dysfunction or altered gait. Mean length of follow-up was 20.6 months (range: 3-68 months). CONCLUSIONS: Despite a high incidence of donor-site and perirectal complications, unilateral functional gluteoplasty was successful in restoring long-term fecal continence in most patients. PMID- 16374100 TI - The use of biomedical sensors to monitor capsule formation around soft tissue implants. AB - Piezoelectric sensors have been shown to respond reproducibly to changes in tissue mechanical properties surrounding an implant over a 4-month period. The vibrational amplitude at a frequency corresponding to the radial resonance shows a statistically significant change over time. The initial period of inflammation is marked by a significant reduction in amplitude, which is indicative of an increase in viscous dissipation of the tissue. As collagen displaces the cellular response, the amplitude continues to decrease. Finally, as the tissue matures, the capsule becomes stiffer, and the viscous dissipation lessens. These results are consistent with qualitative assessments of explanted capsules. Strain gauges encased in a monolithic block of silicone exhibited a greater degree of variability, yet show similar trends over time. The strain increases in the initial 4-week period and remains relatively steady over the following 4 weeks. Beyond 8 weeks, the gauges begin to extrude from the animal or suffer a loss of electrical continuity. Steps are being taken to improve the strain sensor longevity in the animals. PMID- 16374101 TI - Office surgery safety and the Florida moratoria. AB - BACKGROUND: Office-based surgery has become an important method of healthcare delivery, but there is controversy about its safety. Since 2000, a series of articles were published in the lay media emphasizing the hazards of office surgery, leading to the Florida Board of Medicine restricting office procedures. OBJECTIVE: The objective of this study was to determine the nature and scope of deaths resulting from office surgery. METHODS: We reviewed the data on mandatory reporting by physicians to a central agency of all office surgical incidents that resulted in death, injury, or hospital transfer in the state of Florida from January 2000 to November 2004. E-mail, Internet, and telephone follow up were used to determine physician's board status, office accreditation, and hospital privileges. We reviewed data on medication interactions, anesthesia, and monitoring. RESULTS: A total of 36 deaths related to office procedures were reported. Only 18 of those were related to surgical procedures that are within the realm of plastic surgery, although surgeons of other specialties did 3 of these. When these 18 were reviewed by type of anesthesia, there were 12 who had general anesthesia, 10 with an anesthesiologist and 2 with a Certified Registered Nurse Anesthetist. Of those 18, 7 died before discharge. Although all 7 of them survived long enough to be transferred to a hospital, we classified them as office deaths. The other 11 died after appropriate discharge. Of the 7 office deaths, one developed bronchospasm during induction by an anesthesiologist. Five were during deep sedation (level III anesthesia) and 4 appeared to be related to excessive sedation and/or inadequate monitoring; the fifth was probably related to illicit drug use and the sixth from a fat embolism. Of the 11 postoperative deaths, 7 were said to be the result of thromboembolism and the others were from unknown causes. CONCLUSION: Although the total number of office operations during the study period is unknown, the fact that 7 deaths were reported would suggest that the location in which these procedures were done was not as much of a factor as the regulators have suggested. However, better patient screening, sedation management, deep vein thrombosis prophylaxis, and clinical judgment may have prevented some, if not most, of these deaths. The most frequent cause of death after discharge was thromboembolism, and some of these might have been prevented with better prophylaxis. More detailed findings and recommendations are presented. PMID- 16374102 TI - The segmental pectoralis major free flap: anatomical features of its vascular pedicle. AB - BACKGROUND: The pectoralis major muscle may be suited for free transplantation of a segment of the muscle. We investigated the length and diameter of its vascular pedicle to determine its feasibility. METHODS: The length of the pedicle, its arterial diameter, and its entry point into the muscle were determined in 17 cadaveric flaps. RESULTS: The pedicle length up to the medial border of the pectoralis minor muscle averaged 6.6 cm. The mean external arterial diameter was 1.8 mm, and the venous diameter was consistently larger. The vascular pedicle consistently entered the muscle lateral to the midpoint of, and a mean of, 8.8 cm caudal to the clavicular line. CONCLUSION: The vascular length and diameter are sufficient for microvascular anastomosis. Although an anatomic landmark for the cranial border of the flap could not be defined, the sternocostal part of the pectoralis major muscle may potentially be used as a segmental free flap. PMID- 16374103 TI - Outcome analysis, including patient and parental satisfaction, regarding nonvascularized free toe phalanx transfer in congenital hand deformities. AB - The therapy for congenital hand malformations, especially in symbrachydactyly and constriction ring syndromes, is challenging. Between 1975 and 1995, 20 children with congenital hand deformities underwent reconstruction by 56 nonvascularized free toe phalanx transfers. The average age at initial surgery was 4.8 (range, 0.5 to 22) years. Retrospectively, the children were examined after an average of 3.5 (range, 1.5 to 17.6) years for function of the hand, transplanted phalanx growth, assessment for the epiphyseal plate, and assessment for psychologic performance with their parents. Donor-site morbidity was determined according to measured growth deficit, observing the child's gait, and toe function. In the younger patients (up to 1.5 years), the grafts were well tolerated and showed good growth and only few resorptions. In the age group from 1.5 to 4 years, the grafts showed no growth. In the age group older than 4 years, the grafts were mostly resorbed. The clinical reexamination revealed in most cases only passive motion in the joints, but the function of the hand was improved, with only few problems of the donor site. Most patients and their parents reported a positive effect of the phalanx transfer. PMID- 16374104 TI - The human host defense peptide LL37/hCAP accelerates angiogenesis in PEGT/PBT biopolymers. AB - INTRODUCTION: Antimicrobial peptides are effector molecules of the innate immune response and contribute to host defense and inflammation. This study was designed to evaluate neovascularization in biopolymers after instillation with LL37 of angiogenesis in the dorsal skinfold chamber in mice. MATERIAL AND METHODS: The host defense peptide human cathelicicin LL37 was tested for in vitro antimicrobial activity in a bilayer radial diffusion assay. For in vivo testing, 4 different concentrations of LL37 or carrier control were instilled into a biopolymer, then inserted into the dorsal skinfold chamber in Balb/c mice. Standard microcirculatory parameters were assessed over 24 days' follow-up. RESULTS: LL37 showed broad-spectrum antimicrobial activity against gram-positive and -negative bacteria. The LL37 treatment of the biopolymer accelerated the onset of neovascularization by 6 days compared with the carrier control (P < 0.01). CONCLUSION: This study demonstrated that LL37 has antimicrobial activity and is important for early support of neoangiogenesis in biopolymers. The multifunctional human host defense peptide LL37 has potential as an adjunct for tissue engineering. PMID- 16374105 TI - Microsurgical replantation of total upper eyelid avulsion. AB - We present a unique case in which a dog bite resulted in total full-thickness upper eyelid avulsion in a young Caucasian female. The eyelid was replanted using meticulous microsurgical techniques reanastamosing the arterial supply. To our knowledge, this is the first reported case of a successful microvascular replantation of a completely avulsed upper eyelid. The details of the case and the photographic clinical improvement are described and discussed. PMID- 16374106 TI - Discarding the crown jewel. PMID- 16374107 TI - Dorsal slit: an addition to finger dressings. PMID- 16374108 TI - A surgeon's duty in dog bite prevention in children. PMID- 16374109 TI - The mystacial pad flap: a functional facial flap in rats. PMID- 16374110 TI - A versatile, inexpensive permanent marker. PMID- 16374111 TI - Scintigraphic evaluation of intrathecal infusion systems: selection of patients for surgical or medical management. AB - PURPOSE: Continuous intrathecal medication through a surgically implanted infusion system is an established method for treating intractable spasticity or pain. To determine if imaging can guide surgical or medical management in patients with suspected infusion system malfunction, we reviewed our experience in evaluating the functional status of these systems with indium-111 DTPA scintigraphy. METHODS: We retrospectively reviewed 23 indium-111 DTPA studies in 19 patients (7 female, 12 male; ages 20-83 years, average age 47 years). Indications included intractable spasticity or increasing pain. Infusion systems were classified scintigraphically as either functioning or malfunctioning; malfunctions were further classified as either mechanical or catheter-related. Imaging findings were correlated with subsequent clinical management. RESULTS: Fifteen of 23 (65%) studies demonstrated functioning infusion systems; these patients were all treated medically. Eight of 23 (35%) studies demonstrated malfunctioning systems; 6 of 8 were subsequently treated surgically and 2 of 8 medically. CONCLUSION: Indium-111 DTPA scintigraphy can differentiate between functioning and malfunctioning infusion systems, guiding appropriate medical or surgical management. PMID- 16374112 TI - Radiation-induced osteosarcoma and papillary carcinoma of the thyroid. AB - PURPOSE: This article describes a patient with dual radiation-induced malignancies after treatment of neuroblastoma. MATERIALS AND METHODS: A 12-year old boy with a history of neuroblastoma was treated with chemotherapy, I-131 MIBG, and radiotherapy at age 4. He was disease-free for 8 years, but then developed left shoulder pain resulting from osteosarcoma. A thyroid malignancy was discovered during the evaluation. RESULTS: The patient was treated with thyroidectomy and then chemotherapy according to a Children's Cancer Group Study protocol. CONCLUSION: Radiation-induced sarcoma should be suspected whenever a patient who has received radiation several years previously presents with pain or swelling in the irradiated area. PMID- 16374113 TI - The need for intraoperative parathyroid hormone monitoring during radioguided parathyroidectomy by video-assisted thoracoscopy (VATS). AB - PURPOSE: We report on a patient with primary hyperparathyroidism (1HPT) who had a preoperative Tc-99m sestamibi scan localizing a single parathyroid adenoma in the mediastinum. METHODS: On removal of this hyperfunctioning adenoma by radioguided video-assisted thoracoscopic surgery (VATS), intraoperative PTH levels failed to decline in the appropriate manner consistent with curative resection. This prompted the surgical team to investigate further for a second adenoma, which revealed a 2 x 1-cm mass near the inferior border of the thyroid gland on the right lateral aspect of the trachea. RESULTS: In the absence of intraoperative PTH monitoring, the operation would have been terminated after the removal of the mediastinal adenoma, leading to an incomplete surgical resection and persistent 1HPT. CONCLUSION: In our patient, curative resection was obtained and a second operation was avoided because of the use of intraoperative PTH monitoring. This case also emphasizes that although VATS was planned, in treating patients with 1HPT, one must also be prepared to perform a neck exploration. PMID- 16374114 TI - Superiority of F-18 FDG PET imaging for detection of a pheochromocytoma. PMID- 16374115 TI - Pseudolymphoma syndrome: findings on a gallium-67 citrate scan. PMID- 16374116 TI - FDG PET in primary effusion lymphoma (PEL) of the pericardium. PMID- 16374117 TI - False-positive In-111 DTPA octreotide scintigraphy resulting from lung abscess in a patient with metastatic neuroendocrine pancreatic neoplasm. PMID- 16374118 TI - FDG PET imaging of acute cholecystitis. PMID- 16374119 TI - Melioidosis as a cause of multifocal osteomyelitis. PMID- 16374120 TI - F-18 FDG PET superscan. PMID- 16374121 TI - Hip arthroplasty with mass-like pelvic granulomatous disease: PET imaging. PMID- 16374122 TI - Decreased Tc-99m MDP uptake in myelonecrosis. PMID- 16374123 TI - Diffuse metabolic activation of reticuloendothelium on F-18 FDG PET imaging in a case of visceral leishmania. PMID- 16374124 TI - Malignant lesions can mimic gastric uptake on FDG PET. AB - Gastric FDG uptake is a common observation on positron emission tomography (PET), which is most frequently caused by a nonmalignant etiology. However, a malignant lesion in the region of the stomach must not be overlooked. A 48-year-old woman with a history of melanoma in the esophagus and status postesophagectomy underwent FDG PET to evaluate for tumor recurrence. The images demonstrated hypermetabolism in the region of the location of the stomach. However, considering that the patient had a history of a stomach pullthrough procedure, we felt that this activity was nongastric and represented tumor recurrence, which was subsequently confirmed by computed tomography scan and histopathologic examination. PMID- 16374125 TI - Fluorine-18 DOPA PET imaging of paraganglioma syndrome. AB - A 12-year-old boy was admitted after recently having had a resection performed of an extraadrenal retroperitoneal paraganglioma and left adrenalectomy for pheochromocytoma. Despite these procedures, the blood pressure and urinary noradrenalin were elevated. Screening with fluorine-18 DOPA PET demonstrated increased tracer uptake in the right adrenal gland, in a second abdominal lesion, which was prevertebral, and 2 cervical hot spots near the carotid bifurcation, one on each side of the neck. The patient carries a mutation of the gene Succinate dehydrogenasis subunits D (SDHD) and is thus classified with the paraganglioma syndrome type 1. PMID- 16374126 TI - Incidental detection of clinically occult pituitary adenoma on whole-body FDG PET imaging. PMID- 16374127 TI - Tc-99m MDP uptake in retroperitoneal malignant peripheral nerve sheath tumor and its metastases. AB - Extraosseous uptake of Tc-99m MDP has been reported in various pathologic conditions. Because of the opportunity of make an additional diagnosis, extraskeletal distribution of tracer should be inspected in every case. Malignant peripheral nerve sheath tumor (MPNST) is a rare type of neurogenic tumor. Its main clinical manifestation is a mass with or without pain. MPNST can be seen in all 4 extremities, the trunk, head and neck regions, but most commonly occur in the thigh, buttocks, and supraclavicular regions. Presented here is a rare case of MPNST in which Tc-99m MDP uptake by the primary lesion and metastases were clearly demonstrated on bone scintigraphy. PMID- 16374128 TI - Autologous transplant of submandibular gland as a source of tears: demonstration of function on Tc-99m pertechnetate scintigraphy. PMID- 16374129 TI - Fat hypertrophy in a pharyngeal reconstruction flap presenting as an enlarging neck mass: demonstration of benign changes with PET/CT. AB - The patient is a 73-year-old man with a history of radioresistant recurrent squamous cell carcinoma of the larynx, treated with total laryngectomy/pharyngectomy and rectus abdominis deep inferior epigastric perforator flap reconstruction, who presented with a painless enlarging neck mass. Physical examination revealed increased neck bulk and fiberoptic nasopharyngolaryngoscopy was negative. The clinician was concerned about underlying recurrent neoplasm or adenopathy displacing the flap outward and referred the patient for PET/CT. PMID- 16374130 TI - Normal uptake of F-18 FDG in the sublingual gland. PMID- 16374131 TI - Metastatic cancer in thyroid detected by FDG PET. PMID- 16374133 TI - Current readings in nuclear medicine. PMID- 16374132 TI - Indium-111 white blood cell localization in a barium concretion. PMID- 16374134 TI - Prognostic impact of sex on clinical outcomes following emergent coronary angioplasty in acute myocardial infarction. AB - BACKGROUND: Several studies have shown that women had greater risk for adverse clinical outcomes following coronary angioplasty. We aimed to assess the impact of sex on clinical results following emergent coronary angioplasty in acute myocardial infarction. METHODS: We used our database of patients treated for acute myocardial infarction using emergent coronary angioplasty between January 2001 and December 2003. Procedural and angiographic results and clinical outcomes up to 6 months were collected and adjudicated for major cardiac adverse events. The outcome of 352 patients with acute myocardial infarction (71 women, 281 men, no cardiogenic shock) undergoing emergent angioplasty was analyzed and compared according to sex. RESULTS: Acute myocardial infarction occurred at an older age among women who tended to suffer more from diabetes mellitus and hypertension. In addition, on average, women had smaller culprit vessel diameters than men. The immediate post-procedural data were notable for higher frequency of 'no/slow re flow' angiographic phenomenon in women than in men (10.5 vs. 3.4%, P=0.04). In hospital and 30-day mortality was three times higher in women than men (women vs. men: in hospital 7 vs. 2.1%, P=0.05; 30 days 9.9 vs. 3.2%, P=0.02). At 6 months, major adverse cardiac events rate was 28 vs. 15% among women vs. men (P=0.01). Multivariate analysis showed a strong trend towards increased mortality at 30 days among women undergoing acute myocardial infarction angioplasty although it was not significantly or independently related to increased mortality (odds ratio=3.1; confidence intervals=0.8-12.5; P=0.11). CONCLUSION: Our results indicate a trend towards higher early mortality among female patients sustaining acute myocardial infarction and treated using emergent percutaneous coronary intervention that was probably because of increased age and worse coronary flow restoration results among women compared with men. PMID- 16374135 TI - Coronary flow reserve and coronary microvascular functions are strongly related to serum uric acid concentrations in healthy adults. AB - BACKGROUND: Uric acid is a well known antioxidant; however, the relationship between serum uric acid levels and oxidative stress-caused disorders including cardiovascular diseases is not clear yet. Transthoracic Doppler echocardiographic measurement of coronary flow reserve is a useful tool to investigate coronary flow reserve and coronary microvascular functions. In this study, we investigated the possible association between serum uric acid concentrations and coronary flow reserve in healthy adults. METHODS: One hundred healthy volunteers with normal uric acid levels, between 18 and 55 years of age, were included in this study. The study group was divided into two with regard to the serum uric acid levels. Coronary diastolic peak flow velocities were measured at baseline and after dipyridamole infusion (0.56 mg/kg over 4 min) using echocardiography. RESULTS: Coronary flow reserve and hyperemic mean peak flow velocity were significantly greater in participants with lower serum uric acid concentrations (< or =234 micromol/l for women, < or =302 micromol/l for men) than in those with higher serum uric acid concentrations (>234 micromol/l for women, >302 micromol/l for men) (2.91+/-0.5 vs. 2.47+/-0.5, P<0.001; 66.8+/-11.4 vs. 61.1+/-16.5, P=0.04). The baseline mean peak flow velocity was significantly greater in participants with higher serum uric acid concentrations than in those with lower serum uric acid concentrations (24.7+/-4.1 vs. 23.1+/-2.4, P=0.02). CONCLUSION: Lower serum uric acid concentrations might be regarded as indicative of coronary microvascular and conductance vessel functionality. PMID- 16374136 TI - Patient-related variables predicting acute coronary syndrome following admission for chest pain of possible coronary origin. AB - OBJECTIVE: Improving risk stratification of patients experiencing acute chest pain with non-revealing electrocardiogram and cardiac biomarkers could reduce missed acute coronary syndrome and avoid unnecessary hospitalization. METHODS: We assessed the ability of situational, circumstantial, and other patient-related variables in predicting acute coronary syndrome in 921 consecutive patients randomly admitted to this medical department with chest pain of possible coronary origin. A reference group comprised 107 patients referred promptly to the coronary care unit with acute myocardial infarction. RESULTS: Acute coronary syndrome eventually developed in 219 (23.7%) patients. Age and proportions of male patients and those with diabetes, which were significantly lower in the heterogeneous chest pain group than in the reference group, did not differ when re-evaluation was performed between the latter group and the subgroup of patients who eventually developed acute coronary syndrome. Overweight and a family history of premature coronary artery disease remained significantly higher in the reference group, while prevalence of pre-existing coronary artery disease, previous coronary angiography, and coronary intervention remained significantly lower. Variables most significantly predictive of acute coronary syndrome resulted: pre-existing coronary artery disease [odds ratio (OR) 3.2; 95% confidence interval (CI) 2.17-4.71; P<0.001), older age (OR 1.35; 95% CI 1.17 1.57; P<0.001), male sex (OR 1.77; 95% CI 1.19-2.61; P=0.004), diabetes (OR 1.6; 95% CI 1.11-2.32; P=0.01), self-initiation of pain relief treatment before seeking medical help (OR 1.54; 95% CI 1.07-2.23; P=0.02), and conviction that hospitalization for acute coronary disease was mandatory (OR 1.46; 95% CI 1.03 2.07; P=0.03). CONCLUSIONS: Easily obtainable patient-related variables might improve risk stratification and assist physicians to decide on policy in the emergency department and upon hospitalization. PMID- 16374137 TI - Elevated plasma homocysteine levels in patients with isolated coronary artery ectasia. AB - OBJECTIVE: Coronary artery ectasia is a variant of coronary atherosclerosis. Hyperhomocysteinemia has emerged as a major, independent risk factor for cardiovascular diseases. The purposes of this study were to determine plasma hyperhomocysteine levels in patients with coronary artery ectasia, and to compare patients with coronary artery ectasia, coronary artery disease, and controls with normal coronary angiogram. METHOD: The study population included 37 patients with coronary artery ectasia and 36 patients with coronary artery disease. The control group consisted of 32 patients with angiographically proven normal coronary arteries. Plasma hyperhomocysteine levels were measured in all study patients with an enzyme-linked immunosorbent assay. RESULTS: Plasma homocysteine levels were significantly higher in patients with both coronary artery ectasia and coronary artery disease than in the controls (14.8+/-1.1 and 15.9+/-0.8 vs. 2.5+/ 0.6 micromol/l; P<0.001 and P<0.001, respectively). No significant differences in plasma homocysteine levels were found among CAE and CAD groups (P>0.05). CONCLUSIONS: We have demonstrated that patients with coronary artery ectasia and coronary artery disease have increased plasma hyperhomocysteine levels compared with the controls. These findings suggest that hyperhomocysteinemia may play an important role in the pathogenesis of coronary artery ectasia as in coronary artery disease. PMID- 16374138 TI - Decreased endothelium-dependent vasodilatation in patients with migraine: a new aspect to vascular pathophysiology of migraine. AB - BACKGROUND: Migraine is a common neurovascular disorder characterized by attacks of severe headache, autonomic and neurological symptoms. We hypothesized that patients with migraine had abnormal endothelial function. The vascular theory of migraine assumes that the major pathophysiological events that initiate the migraine attack occur in the perivascular nerves of the major cerebral vessels. Accordingly, we aimed to measure endothelium-dependent vasodilatation in migraineurs by means of flow-mediated dilatation, which reflects endothelium dependent vasodilatation capacity. MATERIALS AND METHODS: Forty-five patients who fulfilled the diagnostic criteria for migraine and 45 age and sex-matched healthy participants were enrolled in the study. Flow-mediated dilatation of the brachial artery was determined using a high-resolution B-mode ultrasonographic system. Flow-mediated vasodilatation was expressed as the change in post-stimulus diameter as a percentage of the baseline diameter. RESULTS: Mean ages of the patients were 33+/-10 years in migraineurs (range: 18-52 years, 36 female, 9 male) and 33+/-9 years in non-migraineurs (range: 17-50 years, 36 female and 9 male). Flow-mediated dilatation of patients with migraine is significantly lower than that of the controls (8.02+/-4.095% vs. 10.72+/-3.52%, respectively, P=0.001). CONCLUSION: We have shown that migraineurs have decreased endothelium dependent vasodilatation capacity compared with non-migraineurs. Migraine may be a local manifestation of systemic vascular vasomotion abnormalities. PMID- 16374139 TI - Gene polymorphisms in patients below 35 years of age who underwent coronary artery bypass surgery. AB - OBJECTIVE: Genetic bases for novel prothrombotic, inflammatory risk factors may play a role in the early onset of coronary artery disease. METHODS: Twenty-one patients below 35 years of age who underwent coronary bypass grafting between 2002 and 2004 constituted the study group and were compared with 50 healthy, age and sex-matched controls. Gene analysis for genetic polymorphisms of angiotensin converting enzyme, prothrombin G20210A, tumour necrosis factor-alpha G308A, factor V Leiden and interleukin-6 genes was carried out. RESULTS: The control group was 98% homozygous for the factor V Leiden GG allele and 2% heterozygous for the GA allele. On the other hand, the study group was 76.2% homozygous for the GG allele, and 23.8% heterozygous for the GA allele (P<0.05). Homozygosity for factor V Leiden mutation (AA) was not encountered in either group. With regard to interleukin-6, 70.0% of the control group demonstrated homozygosity for the GG allele and 30.0% showed heterozygosity (GC). The study group was 52.4% homozygous for the GG allele and heterogenicity was similar in this group (28.6% GC). On the other hand, 19.0% of this group demonstrated CC homogenicity (P<0.05). No difference was observed with regard to gene polymorphisms. CONCLUSIONS: Gene polymorphisms with regard to prothrombotic factor V Leiden mutation and inflammatory marker interleukin-6 may play a role in the pathogenesis of early-onset coronary artery stenosis in patients below 35 years of age. PMID- 16374140 TI - Metabolic syndrome negatively impacts early patency of saphenous vein grafts. AB - BACKGROUND: Coronary artery bypass grafting has been performed for a long period utilizing saphenous vein grafts, the fate of which might be crucial to prognosis following the operation. Metabolic syndrome, on the other hand, has become an increasingly important part of cardiovascular practice. We examined whether there was any negative effect of metabolic syndrome on saphenous vein graft patency in a relatively short term (< or =5 years). METHODS: Coronary angiograms of 314 consecutive patients (mean age 62.6+/-8.5 years), having at least one saphenous vein bypass graft within the last 5 years, were evaluated. RESULTS: One hundred and twenty-one patients (group 1) had either an occluded saphenous vein graft or a saphenous vein graft with a significant lesion, and 193 patients (group 2) had patent saphenous vein grafts. Metabolic syndrome was present in 46.2% of all patients (n=145), in 57% of patients in group 1 and in 39.4% of patients in group 2 (P=0.002). Having metabolic syndrome increased the risk of saphenous vein graft occlusion or having a significant lesion on saphenous vein grafts by 2.04-folds. In multivariable logistic regression, smoking (P=0.015, odds ratio=1.88), metabolic syndrome (P=0.019, odds ratio=1.81) and diabetes mellitus (P=0.048, odds ratio=1.36) were found to be associated with poor venous graft fate in the relatively short-term period after bypass. CONCLUSION: Metabolic syndrome seems to impact saphenous vein graft patency negatively in the relatively short term. PMID- 16374141 TI - Increased plasma concentrations of N-terminal pro-brain natriuretic peptide reflect the presence of mildly reduced left ventricular diastolic function in hypertension. AB - BACKGROUND: The potential use of assays of N-terminal pro-brain natriuretic peptide for detection of diastolic abnormalities associated with alterations in blood pressure has not been elucidated. This study was designed to determine whether increased plasma concentrations of N-terminal pro-brain natriuretic peptide sensitively reflect abnormal diastolic function associated with hypertension. METHODS: Concentrations of N-terminal pro-brain natriuretic peptide in plasma were assayed in 40 previously untreated hypertensive patients without overt congestive heart failure and in 20 age and sex-matched controls. Hypertensive patients were studied with the use of pulsed Doppler and color M mode Doppler echocardiography for the evaluation of left ventricular diastolic function. RESULTS: Concentrations of N-terminal pro-brain natriuretic peptide were elevated in hypertensive patients [75.1+/-75.2 (SD) pg/ml compared with 37.9+/-38.5 in controls, P<0.05]. In hypertensive patients, concentrations of N terminal pro-brain natriuretic peptide were negatively correlated with the ratio of color M-mode flow propagation velocity to transmitral E velocity consistent with the view that increased concentrations of N-terminal pro-brain natriuretic peptide are indicative of alterations in diastolic function. Hypertensive patients with N-terminal pro-brain natriuretic peptide values above the mean value in the control group exhibited significantly increased brachial intimal medial thickness and reduced wall stress, consistent with the view that increased N-terminal pro-brain natriuretic peptide was associated with favorable peripheral arterial remodeling. CONCLUSIONS: Elevated concentrations of N-terminal pro-brain natriuretic peptide in plasma reflect the presence of left ventricular diastolic abnormalities and peripheral arterial remodeling in asymptomatic patients with hypertension. PMID- 16374142 TI - Effect of simvastatin on coronary flow reserve in patients with atherosclerosis and hypercholesterolemia: an intracoronary Doppler study. AB - BACKGROUND: Early stages of coronary atherosclerosis are accompanied by a functional impairment of coronary vasodilator capacity and endothelial dysfunction. Reduced coronary flow reserve has been reported in patients with hypercholesterolemia, despite angiographically normal coronary arteries. The aim of the study was to evaluate the effect of simvastatin on coronary flow reserve. METHODS: The study was an open non-placebo-controlled serial investigation in which every patient acted as his own control: 36 male patients with hypercholesterolemia and a non-significant coronary artery lesion in a not previously revascularized coronary artery. Intracoronary Doppler measurements were performed. Coronary flow reserve, relative coronary flow reserve and average peak velocity were performed at baseline, after 3 months on a lipid-lowering diet (control period), and after another 12 months of simvastatin 40 mg/day. In the same patient cohort, significant reduction in lesion plaque plus media has been demonstrated by intravascular ultrasound. RESULTS: Changes in coronary flow reserve were not influenced by either diet or simvastatin (2.5+/-0.6 vs. 2.6+/ 0.5 vs. 2.6+/-0.6, P=ns). Maximum hyperemic flow (34.8+/-12.2 vs. 36.7+/-12.5 vs. 42.5+/-13.1, P<0.001) as well as resting flow (14.3+/-5.3 vs. 14.5+/-4.4 vs. 16.6+/-4.6, P<0.001) increased significantly after 12 months simvastatin therapy. CONCLUSION: Despite plaque plus media, regression simvastatin therapy for 12 months does not affect coronary flow reserve obtained using serial intracoronary Doppler studies. Simvastatin, however, increases the hyperemic flow velocity. PMID- 16374143 TI - Increased ability of tirofiban to maintain its inhibitory effects on the binding of fibrinogen to platelets in blood from patients with and without diabetes mellitus. AB - OBJECTIVES: Both tirofiban and eptifibatide release rapidly from glycoprotein IIb IIIa but have different dissociation constants (KD of tirofiban=15 nmol/l, of eptifibatide=120 nmol/l). Binding of fibrinogen to glycoprotein IIb-IIIa is biphasic, forming an initial reversible complex (KD=155-180 nmol/l) and a second more stable complex (KD=20-70 nmol/l). Diabetes is known to alter platelet function. To determine the influence of affinity on inhibitory effects in blood from patients with (n=20) and without (n=20) diabetes mellitus, we characterized the extent of inhibition as a function of time. METHODS: Blood was added to reaction tubes containing tirofiban 100 ng/ml or eptifibatide 1.7 microg/ml (concentrations previously defined to be optimal) plus a platelet agonist (1 micromol/l adenosine diphosphate or 25 micromol/l thrombin receptor agonist peptide), and fluorochrome-labeled fibrinogen before analysis by flow cytometry. RESULTS: The extent of inhibition early on (30 s to 3 min) was similar (>85%) with either agent in blood from those with and without diabetes mellitus, whereas the extent of inhibition 10-15 min later was maintained more effectively with tirofiban than with eptifibatide (difference in slope P<0.01). After 15 min, the extent of inhibition in response to adenosine diphosphate in those with diabetes mellitus was 95+/-6% for tirofiban and 70+/-15% for eptifibatide (P<0.001); in those without diabetes mellitus, it was 91+/-9% for tirofiban and 73+/-19% for eptifibatide (P<0.001). CONCLUSION: For glycoprotein IIb-IIIa antagonists with a rapid rate of release, the biphasic binding of fibrinogen influences to a similar extent their ability to maintain inhibitory effects in blood from patients with and without diabetes mellitus. PMID- 16374144 TI - Extracorporeal cardiac shock wave therapy ameliorates myocardial ischemia in patients with severe coronary artery disease. AB - OBJECTIVE: Prognosis of severe coronary artery disease with no indication of percutaneous coronary intervention or coronary artery bypass grafting remains poor. We have recently demonstrated that shock wave therapy effectively induces neovascularization and improves myocardial ischemia in a porcine model in vivo. METHODS: With permission from the Ethical Committee of our Institute, we treated nine patients with end-stage coronary artery disease with no indication of percutaneous coronary intervention or coronary artery bypass grafting (55-82 years old, five men and four women) with our cardiac shock wave therapy (200 shots/spot at 0.09 mJ/mm for 20-40 spots, 3 times a week/series). We followed-up the patients at 1, 3, 6, and 12 months after the therapy to examine the amelioration of myocardial ischemia. When needed, shock wave therapy was performed up to three series at 0, and 1, 3 or 6 months. RESULTS: The cardiac shock wave therapy improved symptoms (Canadian Cardiovascular Society functional class score, from 2.7+/-0.2 to 1.8+/-0.2, P<0.01) and reduced nitroglycerin use (from 5.4+/-2.5 to 0.3+/-0.3/week, P<0.05). The treatment also improved myocardial perfusion as assessed by dipyridamole stress thallium scintigraphy (severity score, 25.2+/-7.2% improvement, P<0.05; extent score, 23.3+/-9.0% improvement, P=0.10; washout rate, 20+/-3 to 34+/-3, P<0.05). Myocardial perfusion was improved only in the ischemic area treated with the therapy. These beneficial effects persisted for 12 months. No procedural complications or adverse effects were noted. CONCLUSION: These results indicate that our extracorporeal cardiac shock wave therapy is an effective and non-invasive treatment for end-stage coronary artery disease, although further careful evaluation is needed. PMID- 16374145 TI - Safety of single versus multi-vessel angioplasty for patients with acute myocardial infarction and multi-vessel coronary artery disease: report from the New York State Angioplasty Registry. AB - BACKGROUND: The conventional strategy for primary angioplasty during acute myocardial infarction is angioplasty of the infarct-related vessel, even in patients with multi-vessel disease. Patients, however, often have significant lesions in multiple coronary arteries and a strategy for multi-vessel angioplasty during acute myocardial infarction has not been explored. The purpose of this study was to examine whether multi-vessel angioplasty is as safe as infarct related vessel angioplasty in patients with multi-vessel coronary artery disease during acute myocardial infarction. METHODS: Using the 2000-2001 New York State Angioplasty Registry database, we compared the in-hospital clinical outcomes of patients with multi-vessel disease (>70% stenosis in at least two major coronary arteries), who underwent either multi-vessel angioplasty (n=632) or infarct related vessel angioplasty (n=1350) within 24 h of acute myocardial infarction. Patients with previous myocardial infarction, angioplasty, bypass surgery, or cardiogenic shock were excluded. RESULTS: Patients in the multi-vessel angioplasty group were less likely to be female, to have peripheral vascular disease or diabetes. They had more complex lesions and were more likely to receive stents. In-hospital mortality was three-fold lower (0.8 versus 2.3%, P=0.018) in the multi-vessel angioplasty group. No differences were observed in other ischemic complications, renal failure, or length of stay. After multivariate analysis, multi-vessel angioplasty remained a significant predictor of lower in-hospital death (odds ratio=0.27, 95% confidence interval=0.08-0.90, P=0.03). CONCLUSIONS: Despite the added complexity of multi-vessel angioplasty, patients in this group had significantly lower in-hospital mortality. Therefore, a strategy of multi-vessel angioplasty during acute myocardial infarction may be safe compared with infarct-related angioplasty in selected patients. PMID- 16374146 TI - The metabolic syndrome: a call to action. AB - Metabolic syndrome, a cluster of risk factors that enhances the risk for atherosclerotic cardiovascular disease, has received increasing attention in recent years, especially as the worldwide prevalence of obesity has become better defined. Recent controversy has questioned the scientific basis for metabolic syndrome, but does not negate its value as a description of a common phenotype of patients encountered in clinical practice. Revised and refined diagnostic criteria may be useful for physicians. While more research is needed to understand the pathology of the metabolic syndrome, there is no ambiguity that physicians should treat cardiovascular risk factors in individuals with metabolic syndrome. PMID- 16374147 TI - Bibliography current world literature. PMID- 16374148 TI - Sophorolipids block lethal effects of septic shock in rats in a cecal ligation and puncture model of experimental sepsis. AB - OBJECTIVE: Sophorolipids, a family of natural and easily chemoenzymatically modified microbial glycolipids, are promising modulators of the immune response. The potential of the therapeutic effect of sophorolipids was investigated in vivo in a rat model of sepsis and in vitro by analysis of nitric oxide and cytokine production. DESIGN: Prospective, randomized animal study. SETTING: Experimental laboratory. SUBJECTS: Male Sprague-Dawley rats, 200-240 g. INTERVENTIONS: Intra abdominal sepsis was induced in vivo in 166 rats via cecal ligation and puncture (CLP); 60 rats were used to characterize the model. The remaining rats were treated with sophorolipids or vehicle (dimethylsulfoxide [DMSO]/physiologic saline) by intravenous (iv) tail vein or intraperitoneal (IP) injection immediately post-CLP (25/group). Survival rates were compared at 36 hrs after surgery. In vitro, macrophages were cultured in lipopolysaccharide (LPS) +/- sophorolipid and assayed for nitric oxide (NO) production and gene expression profiles of inflammatory cytokines. In addition, splenic lymphocytes isolated from CLP rats +/- sophorolipid treatment (three per group) were analyzed for cytokine production by RNase protection assay. MEASUREMENTS AND MAIN RESULTS: CLP with 16-gauge needles optimized sepsis induction and resultant mortality. Sophorolipid treatment improved rat survival by 34% (iv) and 14% (IP) in comparison with vehicle controls (p < .05 for iv treatment). Sophorolipids decreased LPS-induced macrophage NO production by 28% (p < .05). mRNA expression of interleukin (IL)-1beta was downregulated by 42.5 +/- 4.7% (p < .05) and transforming growth factor (TGF)-beta1 was upregulated by 11.7 +/- 1.5% (p < .05) in splenocytes obtained 6 hrs postsophorolipid treatment. LPS-treated macrophages cultured 36 hrs with sophorolipids showed increases in mRNA expression of IL 1alpha (51.7%), IL-1beta (31.3%), and IL-6 (66.8%) (p < .05). CONCLUSIONS: Administration of sophorolipids after induction of intra-abdominal sepsis significantly decreases mortality in this model. This may be mediated in part by decreased macrophage NO production and modulation of inflammatory responses. PMID- 16374149 TI - Hypercapnic acidosis and mortality in acute lung injury. AB - OBJECTIVE: We tested the hypothesis that hypercapnic acidosis is associated with reduced mortality rate in patients with acute lung injury independent of changes in mechanical ventilation. DESIGN: Secondary analysis of randomized clinical trial data using hypothesis-driven multivariate logistic regression. SETTING: Randomized, multiple-center trial (n = 861) comparing 12 mL/kg to 6 mL/kg predicted body weight tidal volumes previously published by the National Institutes of Health Acute Respiratory Distress Syndrome (ARDS) Network. PATIENTS: Acute lung injury patients enrolled in a randomized, multiple-center trial (n = 861). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The adjusted odds ratio and 95% confidence intervals (CI) for 28-day mortality rate associated with hypercapnic acidosis defined as day 1 pH <7.35 and Pa(CO2) >45 mm Hg were 0.14 (95% CI 0.03-0.70, p = .016) in the 12 mL/kg predicted body weight tidal volume group and 1.18 (95% CI 0.59-2.35, p = .639) in the 6 mL/kg predicted body weight tidal volume group. Other definitions of hypercapnic acidosis spanning a range of magnitudes suggest a dose-response association between hypercapnic acidosis and 28-day mortality in the 12 mL/kg predicted body weight tidal volume group. None of our definitions of hypercapnic acidosis were associated with reduction in 28-day mortality in the 6 mL/kg predicted body weight tidal volume group. CONCLUSIONS: Hypercapnic acidosis was associated with reduced 28-day mortality in the 12 mL/kg predicted body weight tidal volume group after controlling for comorbidities and severity of lung injury. These results are consistent with a protective effect of hypercapnic acidosis against ventilator associated lung injury that was not found when the further ongoing injury was reduced by 6 mL/kg predicted body weight tidal volumes. PMID- 16374150 TI - Effects of tidal volume on work of breathing during lung-protective ventilation in patients with acute lung injury and acute respiratory distress syndrome. AB - OBJECTIVE: To assess the effects of step-changes in tidal volume on work of breathing during lung-protective ventilation in patients with acute lung injury (ALI) or the acute respiratory distress syndrome (ARDS). DESIGN: Prospective, nonconsecutive patients with ALI/ARDS. SETTING: Adult surgical, trauma, and medical intensive care units at a major inner-city, university-affiliated hospital. PATIENTS: Ten patients with ALI/ARDS managed clinically with lung protective ventilation. INTERVENTIONS: Five patients were ventilated at a progressively smaller tidal volume in 1 mL/kg steps between 8 and 5 mL/kg; five other patients were ventilated at a progressively larger tidal volume from 5 to 8 mL/kg. The volume mode was used with a flow rate of 75 L/min. Minute ventilation was maintained constant at each tidal volume setting. Afterward, patients were placed on continuous positive airway pressure for 1-2 mins to measure their spontaneous tidal volume. MEASUREMENTS AND MAIN RESULTS: Work of breathing and other variables were measured with a pulmonary mechanics monitor (Bicore CP-100). Work of breathing progressively increased (0.86 +/- 0.32, 1.05 +/- 0.40, 1.22 +/- 0.36, and 1.57 +/- 0.43 J/L) at a tidal volume of 8, 7, 6, and 5 mL/kg, respectively. In nine of ten patients there was a strong negative correlation between work of breathing and the ventilator-to-patient tidal volume difference (R = -.75 to -.998). CONCLUSIONS: : The ventilator-delivered tidal volume exerts an independent influence on work of breathing during lung-protective ventilation in patients with ALI/ARDS. Patient work of breathing is inversely related to the difference between the ventilator-delivered tidal volume and patient-generated tidal volume during a brief trial of unassisted breathing. PMID- 16374151 TI - The effect of age on the development and outcome of adult sepsis. AB - OBJECTIVE: Sepsis is an increasingly common and lethal medical condition that occurs in people of all ages. The influence of age on sepsis risk and outcome is incompletely understood. We sought to determine the independent effect of age on the incidence, severity, and outcome of adult sepsis. DESIGN: Longitudinal observational study using national hospital discharge data. SETTING: Approximately 500 geographically separated nonfederal acute care hospitals in the United States. PATIENTS: Patients were 10,422,301 adult sepsis patients hospitalized over 24 yrs, from 1979 to 2002. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Incident sepsis cases were age adjusted and characterized by demographics, sources and types of infection, comorbid medical conditions, and hospital discharge status. Elderly patients (> or = 65 yrs of age) accounted for 12% of the U.S. population and 64.9% of sepsis cases, yielding a relative risk of 13.1 compared with younger patients (95% confidence interval, 12.6-13.6). Elderly patients were more likely to have Gram-negative infections, particularly in association with pneumonia (relative risk, 1.66; 95% confidence interval, 1.63 1.69) and to have comorbid medical conditions (relative risk, 1.99; 95% confidence interval, 1.92-2.06). Case-fatality rates increased linearly by age; age was an independent predictor of mortality in an adjusted multivariable regression (odds ratio, 2.26; 95% confidence interval, 2.17-2.36). Elderly sepsis patients died earlier during hospitalization, and elderly survivors were more likely to be discharged to a nonacute health care facility. CONCLUSIONS: The incidence of sepsis is disproportionately increased in elderly adults, and age is an independent predictor of mortality. Compared with younger sepsis patients, elderly nonsurvivors of sepsis die earlier during hospitalization and elderly survivors more frequently require skilled nursing or rehabilitative care after hospitalization. These findings have implications for patient care and health care resource prioritization and provide insights for expanded scientific investigations and potential patient interventions. PMID- 16374152 TI - Effect of low doses of corticosteroids in septic shock patients with or without early acute respiratory distress syndrome. AB - OBJECTIVE: We investigated the efficacy of low doses of corticosteroids in septic shock patients with or without early acute respiratory distress syndrome (ARDS) by post hoc analysis of a previously completed clinical trial. DESIGN: Retrospective analysis of a placebo-controlled, randomized, double-blind trial of low doses of corticosteroids in septic shock. SETTING: Nineteen intensive care units in France. PATIENTS: Among the 300 septic shock patients enrolled, we selected those meeting standard criteria for ARDS at inclusion. INTERVENTIONS: Seven-day treatment with 50 mg of hydrocortisone every 6 hrs and 50 microg of 9 alpha-fludrocortisone once a day. MEASUREMENTS AND MAIN RESULTS: There were 177 patients with ARDS (placebo, n = 92; corticosteroids, n = 85) including 129 (placebo, n = 67; corticosteroids, n = 62) nonresponders and 48 (placebo, n = 25; corticosteroids, n = 23) responders. In nonresponders, there were 50 deaths (75%) in the placebo group and 33 deaths (53%) in the steroid group (hazard ratio 0.57, 95% confidence interval 0.36-0.89, p = .013; relative risk 0.71, 95% confidence interval 0.54-0.94, p = .011). The number of days alive and off the ventilator was 2.6 +/- 6.6 in the placebo group and 5.7 +/- 8.6 in the steroid group (p = .006). There was no significant difference between groups in responders. There was no significant difference between groups in the two subsets of patients without ARDS. Adverse events rates were similar in the two groups. CONCLUSIONS: This post hoc analysis shows that a 7-day treatment with low doses of corticosteroids was associated with better outcomes in septic shock-associated early ARDS nonresponders, but not in responders and not in septic shock patients without ARDS. PMID- 16374153 TI - Approach to the comatose patient. AB - BACKGROUND: Coma is a medical emergency and may constitute a diagnostic and therapeutic challenge for the intensivist. OBJECTIVE: To review currently available data on the etiology, diagnosis, and outcome of coma. To propose an evidence-based approach for the clinical management of the comatose patient. DATA SOURCE: Search of Medline and Cochrane databases; manual review of bibliographies from selected articles and monographs. DATA SYNTHESIS AND CONCLUSIONS: Coma and other states of impaired consciousness are signs of extensive dysfunction or injury involving the brainstem, diencephalon, or cerebral cortex and are associated with a substantial risk of death and disability. Management of impaired consciousness includes prompt stabilization of vital physiologic functions to prevent secondary neurologic injury, etiological diagnosis, and the institution of brain-directed therapeutic or preventive measures. Neurologic prognosis is determined by the underlying etiology and may be predicted by the combination of clinical signs and electrophysiological tests. PMID- 16374154 TI - Current issues in critical care of the human immunodeficiency virus-infected patient. AB - OBJECTIVE: To provide current information on the epidemiology of human immunodeficiency virus (HIV)-infected patients admitted to the intensive care unit during the era of combination antiretroviral therapy and to review issues related to the administration of antiretroviral therapy that are relevant to the intensivist. DESIGN: Review of literature related to intensive care of HIV infected patients. RESULTS: Overall mortality of HIV-infected patients in the intensive care unit has decreased in the era of combination antiretroviral therapy, and patients are more commonly admitted with non-HIV-related illnesses. Use of antiretroviral therapy in the intensive care unit is difficult but may be associated with improved outcomes. CONCLUSIONS: HIV-infected patients are less likely to be admitted to the intensive care unit with opportunistic infections but more likely to be admitted with problems unrelated to HIV infection or with conditions related to antiretroviral therapy. With current management strategies, more patients survive intensive care unit admission. Intensivists need to be familiar with antiretroviral therapy to recognize life-threatening toxicities unique to these drugs; to avoid drug interactions, which are extremely common and potentially life-threatening; and to avoid enhancing HIV drug resistance, an occurrence that could have devastating consequences for the patient following intensive care unit discharge. PMID- 16374156 TI - Pharmacokinetics and total elimination of meropenem and vancomycin in intensive care unit patients undergoing extended daily dialysis. AB - OBJECTIVE: Extended daily dialysis (EDD) combines the advantage of both intermittent hemodialysis and continuous renal replacement therapy: excellent detoxification accompanied by cardiovascular tolerability. The aim of this study was to evaluate pharmacokinetics of meropenem and vancomycin in critically ill patients with renal failure undergoing EDD. DESIGN: Prospective clinical study. SETTING: Surgical intensive care unit in a tertiary care center. PATIENTS: We studied intensive care patients with anuric acute renal failure being treated with EDD and receiving meropenem (n = 10) or vancomycin (n = 10) therapy. INTERVENTIONS: The antibiotics were administered 6 hrs (1.0 g meropenem) or 12 hrs (1.0 g vancomycin) before EDD was started in order to study the pharmacokinetics before and during EDD. In addition to the application of different methods to calculate pharmacokinetic parameters, the total dialysate concentration of both drugs was measured. RESULTS: Based on the amount of the drug recovered from the collected spent dialysate, the fraction of drug removed by one dialysis treatment was 18% for meropenem and 26% for vancomycin. Dosing regimes for intermittent hemodialysis and continuous renal replacement therapy cannot be used for critically ill patients treated with EDD. CONCLUSION: Our data suggest that patients treated with EDD by means of a high-flux dialyzer (polysulphone; surface area, 1.3 m; blood and dialysate flow, 160 mL/min; EDD time, 480 mins) and current dosing regimens run the risk of being significantly underdosed, which may have detrimental effects on critically ill patients with life-threatening infections. The exact dose has to be tailored according to weight and severity of illness as well as the current minimal inhibitory concentration against the incriminated bacteria. Whenever possible, therapeutic drug monitoring should be performed. PMID- 16374157 TI - Role of nonbronchoscopic lavage for investigating alveolar inflammation and permeability in acute respiratory distress syndrome. AB - OBJECTIVE: Nonbronchoscopic bronchoalveolar lavage is often used as an alternative to bronchoscopic bronchoalveolar lavage in the diagnosis of ventilator-associated pneumonia. We have previously reported an improved safety profile for nonbronchoscopic lavage in patients with lung injury, suggesting that this may be a better technique in this patient group. The objective of this study was to determine whether nonbronchoscopic lavage could be used as an alternative to bronchoscopic lavage for the assessment of alveolar permeability and inflammation in patients at risk for acute respiratory distress syndrome (ARDS) or with ARDS. DESIGN: Prospective randomized crossover trial. PATIENTS: Intubated patients with ARDS or at risk of ARDS. INTERVENTIONS: Bronchoscopic and nonbronchoscopic lavage in the same patient, 15 mins apart. MEASUREMENTS AND MAIN RESULTS: Twenty-one patients with ARDS and 20 patients at risk of ARDS were recruited and underwent nonbronchoscopic and bronchoscopic lavage in randomized order. Despite similar volumes of lavage fluid, nonbronchoscopic lavage had fewer cells and an increased ratio of bronchial epithelial cells to macrophages. Although average concentrations of myeloperoxidase and total protein, the protein permeability index, and the epithelial-lining fluid volume were similar with the two techniques and demonstrated moderate linear associations, Bland and Altman analysis revealed poor comparability, with substantial side-to-side variability and wide 95% limits of agreement. Furthermore, unlike bronchoscopic lavage, nonbronchoscopic lavage was unable to differentiate between patients with ARDS and those at risk of ARDS. CONCLUSIONS: Nonbronchoscopic lavage is not comparable to bronchoscopic lavage and as such cannot be used as an alternative to bronchoscopic lavage for assessing alveolar inflammation in patients with ARDS. PMID- 16374158 TI - Short-stay intensive care after coronary artery bypass surgery: randomized clinical trial on safety and cost-effectiveness. AB - OBJECTIVE: To evaluate the safety and cost-effectiveness of short-stay intensive care (SSIC) treatment for low-risk coronary artery bypass patients. DESIGN: Randomized clinical equivalence trial. SETTING: University Hospital Maastricht, the Netherlands. PATIENTS: Low-risk coronary artery bypass patients. INTERVENTIONS: A total of 600 patients were randomly assigned to undergo either SSIC treatment (8 hrs of intensive care treatment) or control treatment (care as usual, overnight intensive care treatment). MEASUREMENTS: The primary outcome measures were intensive care readmissions and total hospital stay. The secondary outcome measures were total hospital costs, quality of life, postoperative morbidity, and mortality. Hospital costs consisted of the cost of hospital admission or admissions and outpatient costs. MAIN RESULTS: The difference in intensive care readmission between the two groups of 1.13% was very small and not significantly different (p = .241; 95% confidence interval, -0.9% to 2.9%). The total hospital stay (p = .807; 95% confidence interval, 1.2 to -0.4) and postoperative morbidity were comparable between the groups. The SSIC group's quality of life improved more compared with the control group's quality of life (p = .0238; 95% confidence interval, 0.0012 to 0.0464). The total hospital costs for SSIC were significantly lower (95% confidence interval, -1,581 to -174) compared with those for the control group (4,625 and 5,441, respectively). The estimated incremental cost-effectiveness ratio (cost/delta quality-adjusted life months) thus showed the dominance of SSIC. Bootstrap and sensitivity analyses confirm the robustness of the study findings. CONCLUSIONS: Compared with usual care, SSIC is a safe and cost-effective approach. SSIC can be considered as an alternative for conventional postoperative intensive care treatment for low-risk coronary artery bypass graft patients. PMID- 16374159 TI - Initially elevated osteoprotegerin serum levels may predict a perioperative myocardial lesion in patients undergoing coronary artery bypass grafting. AB - OBJECTIVE: We investigated whether osteoprotegerin (OPG), an important regulator in the genesis of arteriosclerosis and bone formation, is able to identify patients at risk for perioperative myocardial infarction measured as cardiac troponin I (cTNI) and signs of myocardial ischemia in the electrocardiogram after coronary artery bypass grafting (CABG). DESIGN: Observational study. SETTING: Post-surgical intensive care unit of a tertiary care center. PATIENTS: Ninety seven patients undergoing elective CABG. INTERVENTIONS: None. MEASUREMENT AND MAIN RESULTS: OPG and cTNI were measured before and 24 hrs after CABG. Additionally, cTNI was measured after 12 hrs. Electrocardiography was done before and immediately after CABG. OPG before CABG (OPGpre) measurements correlated with cTNI measurements after 12 hrs (cTNI12) (r = 0.56; p < .0001) and with cTNI measurements after 24 hrs (cTNI24) (r = 0.77; p < .0001). OPGpre measurements correlated with electrocardiographic findings after surgery (r = 0.65; p < .0001). There was a positive correlation between OPGpre value and the number of bypasses (r = 0.95; p < .0001). A strong correlation was found between OPGpre and homocysteine (r = 0.96; p < .0001). The median OPG presurgical level for the four patients with cardiac complications was found to be notably elevated (28.1 [26.6/31.0] pmol/L) in comparison with that for patients without complications (10.2 [3.7/16.9] pmol/L). CONCLUSIONS: OPG appears to be a useful marker for estimating risk for perioperative myocardial infarction in patients undergoing CABG, as demonstrated by signs of ischemia on electrocardiography. PMID- 16374160 TI - Well-being in informal caregivers of survivors of acute respiratory distress syndrome. AB - OBJECTIVE: With limited community services, the complex rehabilitation period after critical illness is often the responsibility of family members who, as a result, may experience negative health outcomes. The objectives of this research were to a) identify aspects of the caregiving situation that are associated with caregivers' experiences of emotional distress and psychological well-being; and b) compare health-related quality of life of informal caregivers to survivors of acute respiratory distress syndrome (ARDS) with age- and gender-matched population values. DESIGN: Cross-sectional survey of informal caregivers to ARDS survivors. SETTING: Toronto, Ontario, Canada. PATIENTS: Informal caregivers were individuals who were primarily responsible for providing and/or coordinating ARDS survivors' posthospital care and were not paid to do so. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The dependent variables were emotional distress, psychological well-being, and health-related quality of life. They were evaluated by the Center for Epidemiologic Studies Depression Scale, the Positive Affect Scale, and Medical Outcomes Study Short Form 36, respectively. Independent variables included severity of illness indicators, patient depression (Beck Depression Inventory II), aspects of the caregiving experience (care provided, lifestyle interference, personal gain), and psychosocial resources (mastery and social support). Caregivers experienced more emotional distress when they experienced more lifestyle interference, had lower levels of mastery, and were caring for ARDS survivors with more depressive symptoms (F3,42 = 15.69, p < .001, adjusted R = .50). In contrast, caregiver psychological well-being was associated with personal gains as a result of providing care and having more mastery and social support (F4,41 = 9.40, p < .001, adjusted R = .43). Caregivers reported poorer health-related quality of life across all domains of the Medical Outcomes Study Short Form 36 compared with age- and gender-matched population values. CONCLUSIONS: Informal caregivers experience negative health outcomes that persist almost 2 yrs after ARDS. New approaches, such as family-centered rehabilitation, caregiver education, improved respite, and home care, may benefit informal caregivers. PMID- 16374161 TI - Complex analysis of intracranial hypertension using approximate entropy. AB - OBJECTIVE: To determine whether decomplexification of intracranial pressure dynamics occurs during periods of severe intracranial hypertension (intracranial pressure >25 mm Hg for >5 mins in the absence of external noxious stimuli) in pediatric patients with intracranial hypertension. DESIGN: Retrospective analysis of clinical case series over a 30-month period from April 2000 through January 2003. SETTING: Multidisciplinary 16-bed pediatric intensive care unit. PATIENTS: Eleven episodes of intracranial hypertension from seven patients requiring ventriculostomy catheter for intracranial pressure monitoring and/or cerebral spinal fluid drainage. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: We measured changes in the intracranial pressure complexity, estimated by the approximate entropy (ApEn), as patients progressed from a state of normal intracranial pressure (<25 mm Hg) to intracranial hypertension. We found the ApEn mean to be lower during the intracranial hypertension period than during the stable and recovering periods in all the 11 episodes (0.5158 +/- 0.0089, 0.3887 +/- 0.077, and 0.5096 +/- 0.0158, respectively, p < .01). Both the mean reduction in ApEn from the state of normal intracranial pressure (stable region) to intracranial hypertension (-0.1271) and the increase in ApEn from the ICH region to the recovering region (0.1209) were determined to be statistically significant (p < .01). CONCLUSIONS: Our results indicate that decreased complexity of intracranial pressure coincides with periods of intracranial hypertension in brain injury. This suggests that the complex regulatory mechanisms that govern intracranial pressure may be disrupted during acute periods of intracranial hypertension. This phenomenon of decomplexification of physiologic dynamics may have important clinical implications for intracranial pressure management. PMID- 16374162 TI - Predisposing factors for hypoglycemia in the intensive care unit. AB - OBJECTIVE: The introduction of strict glycemic control in the intensive care unit has increased the risk for hypoglycemia. In this study we examined the association between predefined circumstances and the occurrence of hypoglycemia in the intensive care unit. DESIGN: : Retrospective cohort study. SETTING: Academic medical center. PATIENTS: All episodes of hypoglycemia (glucose value <45 mg/dL) in our intensive care unit between September 2002 and September 2004 were identified. Presence of predefined circumstances previously associated with hypoglycemia was scored around the moment of hypoglycemia using a patient data management system and in-hospital charts. Patients with a first hypoglycemic event were contrasted to controls from the same cohort, who were matched for time since admission, to correct for the effect of length of stay. Data were analyzed using conditional logistic regression analysis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of 2,272 patients, 156 (6.9%) experienced at least one episode of hypoglycemia. Continuous venovenous hemofiltration with bicarbonate-based substitution fluid (odds ratio [OR], 14; 95% confidence interval [CI], 1.8-106), a decrease of nutrition without adjustment for insulin infusion (OR, 6.6; 95% CI, 1.9-23), diabetes mellitus (OR, 2.6; 95% CI, 1.5-4.7), insulin use (OR, 5.3; 95% CI, 2.8-11), sepsis (OR, 2.2; 95% CI, 1.2-4.1), and inotropic support (OR, 1.8; 95% CI, 1.1-2.9) were associated with hypoglycemia. Simultaneous octreotide and insulin use (OR, 6.0; 95% CI, 0.72-50) may also be associated with hypoglycemia. Gastric residual during enteral nutrition without adjusting insulin infusion, liver failure, continuous venovenous hemofiltration with lactate-based substitution fluid, diminished glomerular filtration rate, dose diminishment of glucocorticoids or catecholamines, and use of beta-blocking agents were not associated with hypoglycemia. Adjusting for age, gender, and Acute Physiology and Chronic Health Evaluation II score at admission did not materially change ORs. CONCLUSION: Use of bicarbonate-based substitution fluid during continuous venovenous hemofiltration, a decrease of nutrition without adjustment for insulin infusion, a prior diagnosis of diabetes mellitus, sepsis, and need for inotropic support were found to be associated with hypoglycemia. Simultaneous use of insulin and octreotide may be associated with hypoglycemia. PMID- 16374163 TI - Differential diagnostic value of procalcitonin in surgical and medical patients with septic shock. AB - OBJECTIVE: To assess whether different diagnostic and prognostic cutoff values of procalcitonin should be considered in surgical and in medical patients with septic shock. DESIGN: Prospective observational study. SETTING: Intensive care unit of the Avicenne teaching hospital, France. PATIENTS: All patients with septic shock or noninfectious systemic inflammatory response syndrome within 48 hrs after admission. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Patients were allocated to one of the following groups: group 1 (surgical patients with septic shock), group 2 (surgical patients with noninfectious systemic inflammatory response syndrome), group 3 (medical patients with septic shock), and group 4 (medical patients with noninfectious systemic inflammatory response syndrome). Procalcitonin at study entry was compared between group 1 and group 2 and between group 3 and group 4 to determine the diagnostic cutoff value in surgical and in medical patients, respectively. Procalcitonin was compared between survivors and nonsurvivors from group 1 and group 3 to determine its prognostic cutoff value. One hundred forty-three patients were included: 31 in group 1, 36 in group 2, 36 in group 3, and 40 in group 4. Median procalcitonin levels (ng/mL [interquartile range]) were higher in group 1 than in group 3 (34.00 [7.10-76.00] vs. 8.40 [3.63-24.70], p = .01). In surgical patients, the best diagnostic cutoff value was 9.70 ng/mL, with 91.7% sensitivity and 74.2% specificity. In medical patients, the best diagnostic cutoff value was 1.00 ng/mL, with 80% sensitivity and 94% specificity. Procalcitonin was a reliable early prognostic marker in medical but not in surgical patients with septic shock. A cutoff value of 6.00 ng/mL had 76% sensitivity and 72.7% specificity for separating survivors from nonsurvivors. CONCLUSIONS: The diagnostic cutoff value of procalcitonin was higher in surgical than in medical patients. Early procalcitonin was of prognostic interest in medical patients. PMID- 16374164 TI - Microvascular effects following treatment with polyethylene glycol-albumin in lipopolysaccharide-induced endotoxemia. AB - OBJECTIVE: To determine whether resuscitation with polyethylene glycol conjugated bovine serum albumin (2.5% weight/volume) infused at 16 mL/kg/hr (PEG-BSA-16) or at 24 mL/kg/hr (PEG-BSA-24) for 1 hr improves microcirculatory conditions in endotoxemia compared with dextran 70 (6% weight/volume) infused at 24 mL/kg/hr (Dex). DESIGN: Prospective study. SETTING: University research laboratory. SUBJECTS: Male Golden Syrian hamsters. INTERVENTIONS: Hamsters implemented with a skinfold window chamber were given an intravenous injection of lipopolysaccharide and resuscitated within 10 mins with Dex, PEG-BSA-16, or PEG-BSA-24. MEASUREMENTS AND MAIN RESULTS: Hamsters were observed during 24 hrs after lipopolysaccharide injection. Systemic variables measured included mean arterial pressure, heart rate, and systemic arterial blood gas. Microvascular function was characterized by measuring vessel diameter; red blood cell velocity; functional capillary density (FCD); P(O2) in arterioles, venules, and tissue; and perivascular nitric oxide concentration 6 hrs after lipopolysaccharide injection. At 6 hrs, animals with no treatment had the lowest FCD (6.7 +/- 5.7% of baseline). PEG-BSA provided significantly improved microvascular conditions as shown by restoration of FCD. Recovery of FCD was related to improved microvascular flow and perivascular and tissue P(O2), normalization of shear rate, and decreased perivascular nitric oxide concentration. These effects were related to improved fluid retention using PEG-BSA-24 as evidenced by the significantly lower hematocrit at 24 hrs after resuscitation. Nitric oxide at 6 hrs after induction of sepsis achieved perivascular millimolar concentrations, which were reduced to normal values by PEG-BSA-24 treatment. At 6 hrs there were significant differences in FCD, tissue P(O2), and perivascular nitric oxide concentration following PEG-BSA treatment by comparison with Dex treatment, although there were no differences in systemic variables between Dex and PEG-BSA groups. CONCLUSIONS: PEG-BSA produces improved microcirculatory conditions in the treatment of endotoxemia when compared with dextran 70. PMID- 16374165 TI - Free arachidonic versus eicosapentaenoic acid differentially influences the potency of bacterial exotoxins to provoke myocardial depression in isolated rat hearts. AB - OBJECTIVE: Staphylococcal alpha-toxin and Escherichia coli hemolysin (ECH) evoke cardiac dysfunction in isolated rat hearts by provoking myocardial synthesis of arachidonic acid-derived thromboxane A2 or the cysteinyl-leukotrienes, LTC4, LTD4, and LTE4, respectively. We investigated whether low doses of either toxin, which fail to induce cardiac depression by themselves, induce cardiac dysfunction when combined with free arachidonic acid. DESIGN: Prospective, experimental study. SETTING: Research laboratory at a university hospital. SUBJECTS: Isolated hearts from male Wistar rats. INTERVENTIONS: Hearts were perfused with low doses of ECH or alpha-toxin in the absence or presence of arachidonic acid or the alternative eicosanoid precursor eicosapentaenoic acid (EPA). MEASUREMENTS AND MAIN RESULTS: Application of low-dose ECH with arachidonic acid increased coronary perfusion pressure, depressed left ventricular contractile function, provoked electrical instability, and induced a release of creatine kinase concomitant with the liberation of LTC4, LTD4, and LTE4 into the perfusate. All events were abolished when formation of cysteinyl-leukotrienes was blocked by the 5-lipoxygenase activity inhibitor MK-886, targeting 5-lipoxygenase activating protein. In the presence of arachidonic acid, low doses of alpha-toxin caused an increase in cerebral perfusion pressure and a decline of contractile performance, attributable to the release of thromboxane A2, as both events were mitigated by the cyclooxygenase-inhibitor indomethacin. High doses of ECH caused cardiac dysfunction even in the absence of arachidonic acid. However, in the presence of EPA, the cardiodepressant effect of ECH was blunted. Release of EPA-derived LTE5 at the expense of arachidonic acid-derived LTC4, LTD4, and LTE4 was noted in these hearts. CONCLUSIONS: The potency of the bacterial exotoxins ECH and alpha toxin to cause coronary vasoconstriction and myocardial depression is dependent on the availability of free arachidonic acid and may be influenced by supplying omega-3 fatty acids as alternative lipid precursors. PMID- 16374166 TI - Endotoxin-induced heart dysfunction in rats: assessment of myocardial perfusion and permeability and the role of fluid resuscitation. AB - OBJECTIVE: The pathophysiology of sepsis-induced myocardial dysfunction is still controversial. Whether microcirculatory hypoperfusion together with capillary leakage can occur in the heart wall also remains a matter of debate. The objective was to evaluate the impact of fluid resuscitation on endotoxin-induced myocardial dysfunction. DESIGN: Adult rats were given intraperitoneal injection of endotoxin (lipopolysaccharide, Escherichia coli, 10 mg/kg) or phosphate buffered solution, followed up by echocardiography and acetate micro-positron emission tomography scan imaging, together with final hemodynamic, biochemical, and pathologic evaluations up to 48 hrs. SETTING: University laboratory. SUBJECTS: Pathogen-free male Wistar rats (350 g). INTERVENTIONS: Influence of isovolumic fluid infusion type (saline vs. pentastarch) on these variables was assessed in 11 groups of six animals including an unchallenged control one. MEASUREMENTS AND MAIN RESULTS: Endotoxin injection induced a) myocardial dysfunction (decrease of approximately 15-20% in left ventricular ejection fraction); b) ventricular enlargement (approximately 1.5- to 1.7-fold increase in left ventricular systolic volume); c) cardiac output increase (10-15%); d) myocardial hypoperfusion ( approximately 1.5- to 2-fold decrease in acetate k1 constant rate); e) increased oxygen consumption (k2); and f) interstitial wall increase. Endotoxin injection also enhanced levels of arterial lactates and troponin I. Colloid (pentastarch) over crystalloid (saline) fluid resuscitation significantly reversed echocardiographic changes, some positron emission tomography imaging alterations, and lactate and troponin I levels without further enhancing interstitial spaces. CONCLUSION: Endotoxin can induce reversible myocardial alterations with evidence of coronary hypoperfusion and heart wall enlargement/damage, some of which can be prevented by fluid resuscitation. The use of crystalloids is less beneficial than pentastarch. PMID- 16374167 TI - Lack of phosphoinositide 3-kinase-gamma attenuates ventilator-induced lung injury. AB - OBJECTIVE: G protein-coupled receptors may up-regulate the inflammatory response elicited by ventilator-induced lung injury but also regulate cell survival via protein kinase B (Akt) and extracellular signal regulated kinases 1/2 (ERK1/2). The G protein-sensitive phosphoinositide-3-kinase gamma (PI3Kgamma) regulates several cellular functions including inflammation and cell survival. We explored the role of PI3Kgamma on ventilator-induced lung injury. DESIGN: Prospective, randomized, experimental study. SETTING: University animal research laboratory. SUBJECTS: Wild-type (PI3Kgamma), knock-out (PI3Kgamma ), and kinase-dead (PI3Kgamma) mice. INTERVENTIONS: Three ventilatory strategies (no stretch, low stretch, high stretch) were studied in an isolated, nonperfused model of acute lung injury (lung lavage) in PI3Kgamma, PI3Kgamma, and PI3Kgamma mice. MEASUREMENTS AND MAIN RESULTS: Reduction in lung compliance, hyaline membrane formation, and epithelial detachment with high stretch were more pronounced in PI3Kgamma than in PI3Kgamma and PI3Kgamma (p < .01). Inflammatory cytokines and IkBalpha phosphorylation with high stretch did not differ among PI3Kgamma, PI3Kgamma, and PI3Kgamma. Apoptotic index (terminal deoxynucleotidyl transferase mediated biotin-dUTP nick-end labeling) and caspase-3 (immunohistochemistry) with high stretch were larger (p < .01) in PI3Kgamma and PI3Kgamma than in PI3Kgamma. Electron microscopy showed that high stretch caused apoptotic changes in alveolar cells of PI3Kgamma mice whereas PI3Kgamma mice showed necrosis. Phosphorylation of Akt and ERK1/2 with high stretch was more pronounced in PI3Kgamma than in PI3Kgamma and PI3Kgamma (p < .01). CONCLUSIONS: Silencing PI3Kgamma seems to attenuate functional and morphological consequences of ventilator-induced lung injury independently of inhibitory effects on cytokines release but through the enhancement of pulmonary apoptosis. PMID- 16374168 TI - Thermal injury-induced peroxynitrite production and pulmonary inducible nitric oxide synthase expression depend on JNK/AP-1 signaling. AB - OBJECTIVE: To determine whether burn-induced peroxynitrite production and expression of lung inducible nitric oxide synthase (iNOS), intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, CXCR2, macrophage inflammatory protein (MIP)-2, and neutrophil chemokine (KC) are mediated by the c Jun NH2-terminal kinase (JNK). DESIGN: Prospective, experimental study. SETTING: Research laboratory at a university hospital. SUBJECTS: Thermal injury models in the mice. INTERVENTIONS: In experiment 1, specific pathogen-free C57/BL6 mice were subjected to 30% total body surface area third-degree burn over shaved back. At 0 hr, 2 hrs, 4 hrs, and 6 hrs after burn, lung tissues of those mice were harvested for JNK activity assay, AP-1 DNA-binding activity, and pJNK immunohistochemistry. In experiment 2, a specific JNK inhibitor, SP600125, was given (30 mg/kg intraperitoneally) to mice immediately postburn to suppress the JNK activity. At 8 hrs after burn, blood was assayed for the peroxynitrite mediated dihydrorhodamine (DHR) 123 oxidation. Lung tissues were harvested for myeloperoxidase (MPO) determination, ICAM-1, VCAM-1, CXCR2, KC, MIP-2, interleukin-1beta, and interleukin-6 messenger RNA expression; iNOS immunohistochemical staining; and histologic studies. Pulmonary microvascular dysfunction was quantified by measuring the extravasations of Evans blue dye. MEASUREMENTS AND MAIN RESULTS: The JNK activity and AP-1 DNA-binding activity of lung tissue significantly increased to a peak at 2 hrs and 4 hrs, respectively, after thermal injury. Immunohistochemical study demonstrated that the increase of the pJNK was mostly from the bronchiole epithelial cells. This increase of MPO activity in lung, blood DHR 123 oxidation level, and lung permeability increased six-fold, nine-fold, and four-fold after burn. SP600125 administration obliterated the thermal injury-induced JNK activity, AP-1 DNA-binding activity, and iNOS expression in lung tissue. SP600125 treatment also significantly decreased MPO activity, blood DHR 123 oxidation, and lung permeability by 54%, 8%, and 47%, respectively, and markedly decreased the thermal injury-induced perivascular and interstitial inflammatory cell infiltration and septum edema. Furthermore, SP600125 abolished thermal injury-induced ICAM-1, VCAM-1, CXCR2, MIP 2, and KC but not interleukin-1beta and interleukin-6 messenger RNA levels of lung tissues. CONCLUSIONS: Thermal injury induces lung tissue JNK activation and AP-1 DNA-binding activity mainly from airway epithelial cells. Thermal injury induced peroxynitrite production and lung iNOS, ICAM-1, and VCAM-1 expression are mediated by the JNK signaling. JNK inhibition decreases thermal injury-induced lung neutrophil infiltration and subsequently pulmonary hyperpermeability. PMID- 16374169 TI - Simulation-based training is superior to problem-based learning for the acquisition of critical assessment and management skills. AB - OBJECTIVE: To determine whether full-scale simulation (SIM) is superior to interactive problem-based learning (PBL) for teaching medical students acute care assessment and management skills. DESIGN: Randomized controlled trial. SETTING: Simulation center at a U.S. medical school. SUBJECTS: Thirty-one fourth-year medical students in a week-long acute care course. INTERVENTIONS: After institutional review board approval and informed consent, eligible students were randomized to either the SIM or PBL group. On day 1, all subjects underwent a simulator-based initial assessment designed to evaluate their critical care skills. Two blinded investigators assessed each student using a standardized checklist. Subsequently, the PBL group learned about dyspnea in a standard PBL format. The SIM group learned about dyspnea using the simulator. To equalize simulator education time, the PBL group learned about acute abdominal pain on the simulator, whereas the SIM group used the PBL format. On day 5, each student was tested on a unique dyspnea scenario. MEASUREMENTS AND MAIN RESULTS: Mean initial assessment and final assessment checklist scores and their change for the SIM and PBL groups were compared using the Student's t-test. A p < .05 was considered significant. The SIM and PBL groups had similar mean (PBL 0.44, SIM 0.47, p = .64) initial assessment scores (earned score divided by maximum score) and were deemed equivalent. The SIM group performed better than the PBL group on the final assessment (mean, PBL 0.53, SIM 0.72, p < .0001). When each student's change in score (percent correct on final assessment minus percent correct on the initial assessment) was compared, SIM group students performed better (mean improvement, SIM 25 percentage points vs. PBL 8 percentage points, p < .04) CONCLUSIONS: For fourth-year medical students, simulation-based learning was superior to problem based learning for the acquisition of critical assessment and management skills. PMID- 16374170 TI - Effect of albumin dialysis on intracranial pressure increase in pigs with acute liver failure: a randomized study. AB - BACKGROUND: Increased intracranial pressure (ICP) worsens the outcome of acute liver failure (ALF). This study investigates the underlying pathophysiological mechanisms and evaluates the therapeutic effect of albumin dialysis in ALF with use of the Molecular Adsorbents Recirculating System without hemofiltration/dialysis (modified, M-MARS). METHODS: Pigs were randomized into three groups: sham, ALF, and ALF + M-MARS. ALF was induced by hepatic devascularization (time = 0). M-MARS began at time = 2 and ended with the experiment at time = 6. ICP, arterial ammonia, brain water, cerebral blood flow (CBF), and plasma inflammatory markers were measured. RESULTS: ICP and arterial ammonia increased significantly over 6 hrs in the ALF group, in comparison with the sham group. M-MARS attenuated (did not normalize) the increased ICP in the ALF group, whereas arterial ammonia was unaltered by M-MARS. Brain water in the frontal cortex (grey matter) and in the subcortical white matter at 6 hrs was significantly higher in the ALF group than in the sham group. M-MARS prevented a rise in water content, but only in white matter. CBF and inflammatory mediators remained unchanged in all groups. CONCLUSION: The initial development of cerebral edema and increased ICP occurs independently of CBF changes in this noninflammatory model of ALF. Factor(s) other than or in addition to hyperammonemia are important, however, and may be more amenable to alteration by albumin dialysis. PMID- 16374171 TI - Beta2-adrenergic agonist protects human endothelial cells from hypoxia/reoxygenation injury in vitro. AB - OBJECTIVE: Circulatory shock results in hypoxia/reoxygenation processes that lead to the release of reactive oxygen species, endothelial injury, and multiple organ failure. Previous data suggest that beta2-adrenergic agonists prevent endothelial dysfunction. The study aimed at determining whether the beta2-adrenergic agonist formoterol protects endothelial cells against hypoxia/reoxygenation injury in vitro. DESIGN: Prospective controlled trial. SETTING: University hospital research laboratory. SUBJECTS: Cultured human umbilical vein endothelial cells (HUVECs). INTERVENTIONS: Confluent HUVECs were sealed in a flow-through chamber mounted on an inverted microscope and perfused with a constant flow of Krebs medium. After 1 hr of equilibration, HUVECs underwent 2 hrs of hypoxia and 1 hr of reoxygenation. Cell death at the end of reoxygenation and reactive oxygen species formation were assessed with fluorescent probes propidium iodide and 2',7'-dichlorodihydrofluorescein diacetate, respectively. The effects of the beta2-adrenergic agonist formoterol, the beta2-adrenergic antagonist ICI 118,551 and the nitric oxide synthase inhibitor L-NNA were investigated. Statistical analysis was performed with analysis of variance followed by post hoc Fisher's test. MEASUREMENTS AND MAIN RESULTS: Hypoxia/reoxygenation increased cell death (hypoxia/reoxygenation 29 +/- 4% vs. control 1 +/- 5%, p < .05) and endothelial reactive oxygen species production (hypoxia/reoxygenation 126 +/- 4% vs. control 108 +/- 4%, p < .05). Formoterol reduced cell death in a concentration-dependent manner (EC95 = 10 mol/L) and reduced endothelial reactive oxygen species production (hypoxia/reoxygenation + formoterol EC95 109 +/- 4% vs. hypoxia/reoxygenation 126 +/- 4%, p < .05). When added to formoterol EC95, ICI 118,551 and L-NNA abolished the formoterol-induced cell protection and reduced reactive oxygen species production. CONCLUSIONS: These results indicate that formoterol reduces endothelial cell death and reactive oxygen species production in this in vitro hypoxia/reoxygenation model. These effects are beta2-adrenergic specific and are partially mediated by nitric oxide synthase. PMID- 16374172 TI - Nitric oxide modifies the sarcoplasmic reticular calcium release channel in endotoxemia by both guanosine-3',5' (cyclic) phosphate-dependent and independent pathways. AB - OBJECTIVES: a) To determine whether decreased sarcoplasmic calcium release channel (CRC) activity is a mechanism by which myocardial contractility is reduced in endotoxemia; b) to determine whether nitric oxide modulates CRC activity in endotoxemia; and c) to examine two nitric oxide signaling pathways in relation to CRC function in endotoxemia. DESIGN: Randomized, prospective using a rat model of endotoxemia. SETTING: : Research laboratory. SUBJECTS: Sprague Dawley rats. INTERVENTIONS: Endotoxemia was induced by lipopolysaccharide administration. The effects of nitric oxide were studied using the highly selective inducible nitric oxide synthase inhibitor N-(3 (aminomethyl)benzyl)acetamidine dihydrochloride (1400W) and the specific guanylyl cyclase inhibitor 1-H (1, 2, 4)oxadiazolo[4,3-a]quinoxalin-1-one (ODQ). MEASUREMENTS AND MAIN RESULTS: We assessed myocardial contractility, myocardial nitric oxide content, and guanosine-3',5' (cyclic) phosphate (cGMP) content. We determined CRC activity by calcium release and ryanodine binding assays. We followed these variables at four time points through the course of endotoxemia. We found that myocardial contractility and CRC activity were decreased in late but not in early endotoxemia. Furthermore, inducible nitric oxide synthase inhibition with 1400W restored contractility and CRC activity in late endotoxemia but paradoxically worsened these variables in early endotoxemia. Through the use of the guanylyl cyclase inhibitor ODQ, we demonstrate that nitric oxide acts through cGMP-mediated mechanisms in early and late endotoxemia. We investigated cGMP-independent pathways by assessing the oxidative status of the CRC. We found that in late endotoxemia, nitric oxide decreased the number of free thiols, demonstrating that nitric oxide also acts through cGMP-independent pathways. CONCLUSIONS: Nitric oxide has a dual effect on the CRC in endotoxemia. At low concentrations, as measured in early endotoxemia, nitric oxide stabilizes the CRC through cGMP-mediated mechanisms. In late endotoxemia, high nitric oxide concentrations decrease channel activity through both cGMP-dependent and cGMP independent mechanisms. PMID- 16374173 TI - Gut ischemia-reperfusion affects gut mucosal immunity: a possible mechanism for infectious complications after severe surgical insults. AB - OBJECTIVE: To examine influences of gut ischemia/reperfusion (I/R) on gut associated lymphoid tissue (GALT) mass and function. DESIGN: Prospective, randomized controlled study. SETTING: Research laboratory. SUBJECTS: Male Institute of Cancer Research mice. INTERVENTIONS: Ninety mice were randomized to three groups: I/R (60-min gut ischemia), sham (laparotomy only), and control (no operation). On days 1, 2, 4, 7, and 10, mice were killed to harvest lymphocytes from Peyer patches, the intraepithelial space, and the lamina propria (LP) of the small intestine. Respiratory tract and small intestinal washings were also obtained. MEASUREMENTS AND MAIN RESULTS: Gut I/R significantly reduced lymphocyte numbers in Peyer patches, the intraepithelial space, and the LP. The reduction was prominent in GALT effector sites, that is, the intraepithelial space and LP, but numbers recovered quickly in LP. Changes in cell numbers in Peyer patches, GALT inductive sites, were subtle but persistent. Gut I/R reduced B cell numbers in Peyer patches; alphabeta T cell receptor (TCR)+, gammadeltaTCR+, CD8+, and B cell numbers in the intraepithelial space; and gammadeltaTCR+, CD8+, and B cell numbers in the LP, in comparison with the sham or control group. There were no significant differences in respiratory tract immunoglobulin A levels between the I/R and sham groups. Intestinal immunoglobulin A was elevated on day 1 in the I/R group, with no significant difference after day 2 in comparison with the sham group. CONCLUSIONS: Despite the maintained mucosal immunoglobulin A level, gut I/R markedly reduces GALT cell numbers, with changes in lymphocyte phenotypes. These alterations may be associated with increased morbidity due to infectious complications after severe surgical insults. PMID- 16374174 TI - Acute lung injury in patients with subarachnoid hemorrhage: incidence, risk factors, and outcome. AB - OBJECTIVE: Pulmonary complications account for significant morbidity and mortality following aneurysmal subarachnoid hemorrhage; however, the effect of acute lung injury is largely unknown. The goal of this study was to determine the incidence of acute lung injury in a large cohort of patients with subarachnoid hemorrhage as well as determine the risk factors for acute lung injury and its effect on mortality and length of stay. Ventilator management was analyzed to determine the proportion of patients with subarachnoid hemorrhage and acute lung injury who a received a low-tidal volume ventilation strategy. DESIGN: Retrospective cohort study. SETTING: University-affiliated county hospital in Seattle, WA. PATIENTS: Six-hundred and twenty patients with aneurysmal subarachnoid hemorrhage documented on computed tomography or angiography. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: One-hundred and seventy patients met criteria for acute lung injury (incidence, 27%; 95% confidence interval, 24-31%). On multivariate analysis, severity of illness, clinical grade of hemorrhage, packed red blood cell transfusions, and severe sepsis in the intensive care unit were independently associated with development of acute lung injury. After adjustment for important confounders, development of acute lung injury was associated with a statistically significant increase in hospital mortality (odds ratio, 1.63; 95% confidence interval, 1.03-2.57). Acute lung injury was also independently associated with an increased intensive care unit length of stay (15%, 95% confidence interval, 5-27%). Thirty percent of patients with acute lung injury received low tidal volume ventilation. Patients receiving low tidal volume ventilation had worse oxygenation and higher positive end expiratory pressure requirements compared with those who did not, but there were no significant differences in arterial pH or Pco2. CONCLUSIONS: Acute lung injury is common in patients with subarachnoid hemorrhage and is independently associated with a worse clinical outcome. Research is needed to determine the causes of acute lung injury in this population and whether these patients are candidates for evidence-based ventilator strategies to reduce mortality. PMID- 16374175 TI - Effects of a recombinant surfactant protein-C-based surfactant on lung function and the pulmonary surfactant system in a model of meconium aspiration syndrome. AB - OBJECTIVE: Meconium aspiration syndrome (MAS) remains a relevant cause of neonatal respiratory failure and is characterized by severe impairment of pulmonary gas exchange, surfactant inactivation, and pronounced inflammatory changes. Surfactant administration has been shown as an effective treatment strategy in MAS. The present study aimed at investigating the impact of a recombinant surfactant protein (SP)-C-based surfactant on pulmonary gas exchange and lung function in this model of neonatal lung injury. Furthermore, SP-B and -C were determined on the transcriptional and protein level. DESIGN: Laboratory experiment. SETTING: University laboratory. SUBJECTS: Twenty three newborn piglets (median age 6 days, weight 1900-2500 g). INTERVENTIONS: Piglets were intubated and mechanically ventilated and then received 20% sterile meconium (5 mL/kg) for induction of lung injury. After 30 mins, animals were randomized for control (n = 7, MAS controls), recombinant SP-C surfactant (n = 8), or natural surfactant (n = 8). Surfactant preparations were administered as an intratracheal bolus (75 mg/kg), and animals were ventilated for another 330 mins. Nonventilated newborn piglets at term (n = 28; median weight 1484 g, range 720-1990 g) served as a healthy reference group (healthy controls). MEASUREMENTS AND MAIN RESULTS: Lung function variables, arterial blood gas samples, and lung tissues were obtained. Expression of SP-B and -C messenger RNA was quantified in left lung lobe tissue using real-time polymerase chain reaction. Protein concentrations were determined by enzyme-linked immunosorbent assay. Scanning electron microscopy and transmission electron microscopy were performed in tissue samples of the right lung lobe. Compared with healthy controls, SP-B messenger RNA expression was significantly increased in MAS (p < .02), whereas SP-C messenger RNA expression was found to be significantly reduced (p < .001). SP concentrations, however, were not significantly different. Although a significant improvement of gas exchange and lung function was observed after surfactant administration in both groups, surfactant messenger RNA expression and protein concentrations were not significantly altered. Scanning and transmission electron microscopy showed severe pulmonary ultrastructural changes after meconium aspiration improving after surfactant treatment. CONCLUSIONS: Impairment of lung function in MAS, associated with marked changes in SP messenger RNA expression, can be sufficiently treated using recombinant SP-C-based or natural surfactant. Despite improved lung function and gas exchange as well as pulmonary ultrastructure after treatment, pulmonary SP messenger RNA expression and concentrations remained significantly affected, giving important insight into the time course following surfactant treatment in MAS. PMID- 16374176 TI - Intensive care unit quality improvement: a "how-to" guide for the interdisciplinary team. AB - OBJECTIVE: Quality improvement is an important activity for all members of the interdisciplinary critical care team. Although an increasing number of resources are available to guide clinicians, quality improvement activities can be overwhelming. Therefore, the Society of Critical Care Medicine charged this Outcomes Task Force with creating a "how-to" guide that focuses on critical care, summarizes key concepts, and outlines a practical approach to the development, implementation, evaluation, and maintenance of an interdisciplinary quality improvement program in the intensive care unit. DATA SOURCES AND METHODS: The task force met in person twice and by conference call twice to write this document. We also conducted a literature search on "quality improvement" and "critical care or intensive care" and searched online for additional resources. DATA SYNTHESIS AND OVERVIEW: We present an overview of quality improvement in the intensive care unit setting and then describe the following steps for initiating or improving an interdisciplinary critical care quality improvement program: a) identify local motivation, support teamwork, and develop strong leadership; b) prioritize potential projects and choose the first target; c) operationalize the measures, build support for the project, and develop a business plan; d) perform an environmental scan to better understand the problem, potential barriers, opportunities, and resources for the project; e) create a data collection system that accurately measures baseline performance and future improvements; f) create a data reporting system that allows clinicians and others to understand the problem; g) introduce effective strategies to change clinician behavior. In addition, we identify four steps for evaluating and maintaining this program: a) determine whether the target is changing with periodic data collection; b) modify behavior change strategies to improve or sustain improvements; c) focus on interdisciplinary collaboration; and d) develop and sustain support from the hospital leadership. We also identify a number of online resources to complement this overview. CONCLUSIONS: This Society of Critical Care Medicine Task Force report provides an overview for clinicians interested in developing or improving a quality improvement program using a step-wise approach. Success depends not only on committed interdisciplinary work that is incremental and continuous but also on strong leadership. Further research is needed to refine the methods and identify the most cost-effective means of improving the quality of health care received by critically ill patients and their families. PMID- 16374177 TI - Elevated growth-arrest-specific protein 6 plasma levels in patients with severe sepsis. AB - OBJECTIVE: Growth-arrest-specific protein 6 (Gas6), an intracellular protein released by apoptotic cells, has been detected in normal plasma. As the Gas6 system has been implicated in mouse susceptibility to sepsis, and as leukocyte apoptosis is thought to play a major role in the physiopathology of human severe sepsis, we studied Gas6 plasma levels and possibly related variables in patients with severe sepsis. DESIGN: Matched case-control study. SETTING: Adult intensive care unit in a university hospital. PATIENTS: Thirty patients with severe sepsis, 30 patients with organ failure not related to infection, and 30 healthy subjects matched for age and gender. INTERVENTIONS: Blood draw. MEASUREMENTS AND MAIN RESULTS: Gas6 plasma levels were quantified using enzyme-linked immunosorbent assay. Whole-blood gas6 messenger RNA levels were measured by quantitative real time polymerase chain reaction. Gas6 plasma levels were elevated (110 ng/mL [75, 139]; median values [interquartile range]) in severe sepsis patients compared with organ failure patients (85 ng/mL [56, 101]) and healthy subjects (54 ng/mL [49, 68]). In patients with severe sepsis, this increase correlated with the Acute Physiology and Chronic Health Evaluation II severity score, the organ failure Organ Dysfunction and Infection (ODIN) score, and the existence of a septic shock. Gas6 messenger RNA levels were increased in patients with severe sepsis and correlated specifically with the monocyte count. CONCLUSIONS: In severe sepsis, the recently described anti-apoptotic protein Gas6 was found at high levels in plasma and correlated well with the degree of organ dysfunction. PMID- 16374178 TI - Pressure-flow signatures of central-airway mucus plugging. AB - SETTING: Medical Intensive Care Unit of Regions Hospital, a University of Minnesota-affiliated teaching hospital. PATIENT: Mechanically ventilated woman with status asthmaticus and acute respiratory failure. INTERVENTION: Observations of airway pressure and flow tracings before and after bronchoscopic inspection and airway lavage. MAIN RESULTS: Four newly observed signs were recorded that may serve to identify occult central airway mucus plugging in the ventilated asthmatic patient. PMID- 16374179 TI - Finding out what we do in the ICU. PMID- 16374180 TI - Respiratory acidosis and acute respiratory distress syndrome: time to trade in a bull market? PMID- 16374181 TI - Patient-ventilator dyssynchrony during lung protective ventilation: what's a clinician to do? PMID- 16374182 TI - Sepsis in an aging population. PMID- 16374183 TI - Effectiveness of prolonged glucocorticoid treatment in acute respiratory distress syndrome: the right drug, the right way? PMID- 16374184 TI - Out of intense complexities, intense simplicities emerge: AIDS in the intensive care unit. PMID- 16374185 TI - Filtering out important considerations for developing drug-dosing regimens in extended daily dialysis. PMID- 16374186 TI - Grading the evidence for shortened intensive care unit stays after cardiac operations. PMID- 16374187 TI - Focusing on caregivers of the critically ill: beyond illness into recovery. PMID- 16374188 TI - Phenomenological issues related to the measurement, mechanisms and manipulation of complex biological systems. PMID- 16374189 TI - Intensive insulin therapy: of harm and health, of hypes and hypoglycemia. PMID- 16374190 TI - The Holy Grail of volume resuscitation in the septic patient is... PMID- 16374191 TI - Phosphoinositide-3-kinase gamma: is it really pivotal for ventilator-induced lung injury? PMID- 16374192 TI - Clinical skills in acute care: a role for simulation training. PMID- 16374193 TI - Drain the brain: albumin dialysis for intracranial hypertension. PMID- 16374194 TI - The role of nitric oxide signaling in sepsis-induced myocardial dysfunction. PMID- 16374195 TI - Linking gut-associated lymphoid tissue to multiple organ dysfunction syndrome and infection. PMID- 16374196 TI - Sophorolipids in sepsis: antiinflammatory or antibacterial? PMID- 16374197 TI - Different therapeutic perspectives for novel exogenous surfactant preparations. PMID- 16374198 TI - Quality improvement: best evidence in clinical practice and clinical evidence of best practice. PMID- 16374199 TI - Tidal volumes in acute respiratory distress syndrome--one size does not fit all. PMID- 16374201 TI - An excellent pharmacokinetic profile can actually act as a double-edged sword. PMID- 16374202 TI - Raising mean arterial pressure in patients with septic shock. PMID- 16374205 TI - Draining all pleural effusions in the intensive care unit? PMID- 16374207 TI - Serum lipids or apolipoprotein A-I and disease severity of meningococcal sepsis. PMID- 16374210 TI - HIV-1 subtypes: epidemiology and significance for HIV management. AB - PURPOSE OF REVIEW: This review presents an update on the molecular epidemic patterns of HIV-1 infection and the effects of subtype-related genetic variability on transmission, disease progression, response to antiretroviral therapy and drug-resistance pathways. RECENT FINDINGS: The molecular epidemiology of HIV-1 infection is complex and evolving. The emergence of new variants reflects HIV-1 prevalence, subtype epidemiology and risk-behaviour patterns in different geographical areas. Evidence indicates that certain subtypes may have a transmission advantage while others display higher replicative efficiency. The molecular mechanisms underlying these differences are being identified and include both virus- and host-related factors. Although drug susceptibility varies and clinical evidence remains limited, current antiretroviral regimens appear to have comparable efficacy in patients infected with B and non-B subtypes. Subtype related variability influences resistance pathways. However, the major treatment associated resistance mutations seen in subtype B also confer resistance in non-B subtypes and vice versa. SUMMARY: Genetic differences among HIV-1 variants can influence the virus biological properties, susceptibility to existing and candidate antiretroviral drugs, and evolution of antiretroviral drug resistance. Further studies are required to define the impact of this variability on risk of transmission, disease outcomes, responses to antiretroviral therapy and resistance pathways. Meanwhile, plasma viral load and CD4 count remain the important predictors of disease outcome, regardless of the infecting subtype. Current antiretroviral regimens can be used reliably to treat patients with both B and non-B subtypes, and resistance interpretation algorithms provide adequate guidance. The limitations of current evidence should be acknowledged and instigate ongoing vigilance. PMID- 16374211 TI - Management of hepatitis C/HIV coinfection. AB - PURPOSE OF REVIEW: One third of HIV-infected individuals in Europe and the USA have a hepatitis C coinfection. With the introduction of highly active antiretroviral therapy for treatment of HIV, liver disease caused by chronic hepatitis C virus infection has now become an increasingly important cause of morbidity and mortality among HIV-infected patients. Therefore, treatment strategies for management of hepatitis C coinfection in HIV-infected individuals are urgently needed. RECENT FINDINGS: With the introduction of pegylated interferon/ribavirin combination therapy significantly improved treatment options for HIV/hepatitis C virus-coinfected patients have become available, leading to sustained virological response rates of over 40%. Increasing knowledge on the management of adverse events under hepatitis C therapy and optimized selection of antiretrovirals in HIV/hepatitis C virus-coinfected patients has helped to reduce complications and improve overall treatment outcome. SUMMARY: Treatment with pegylated interferon plus ribavirin is safe and effective in HIV/hepatitis C virus-coinfected patients. Longer treatment durations of 48 weeks are recommended for genotype 2 or 3. Positive predictive factors for sustained response are hepatitis C virus genotype 2 or 3 and early treatment response. PMID- 16374212 TI - AIDS-related malignancies: changing epidemiology and the impact of highly active antiretroviral therapy. AB - PURPOSE OF REVIEW: Three cancers in people with HIV denote an AIDS diagnosis: Kaposi's sarcoma, high-grade B-cell non-Hodgkin's lymphoma and invasive cervical cancer. In addition a number of other cancers occur at increased frequency in this population group but are not AIDS-defining illnesses. This review discusses the impact of highly active antiretroviral therapy on the epidemiology and outcome of AIDS-defining cancers. RECENT FINDINGS: The incidence of both Kaposi's sarcoma and non-Hodgkin's lymphoma has declined in the era of highly active antiretroviral therapy and the outcome of both tumours has improved. Moreover, highly active antiretroviral therapy alone produces a response in a majority of antiretroviral-naive patients with Kaposi's sarcoma. In contrast, highly active antiretroviral therapy has had little impact on the incidence of human papilloma virus-associated tumours (cervical and anal cancer) in people with HIV, although it may improve survival by reducing opportunistic infection deaths. As people with HIV live longer with highly active antiretroviral therapy, an increased incidence of other non AIDS-defining cancers that have no known association with oncogenic infections is becoming apparent. SUMMARY: For those with access to highly active antiretroviral therapy, the good news from the AIDS-defining cancers - particularly Kaposi's sarcoma and non-Hodgkin's lymphoma - may be balanced by the increasing numbers of non AIDS-defining cancers. PMID- 16374213 TI - Immune reconstitution inflammatory syndrome in HIV. AB - PURPOSE OF REVIEW: The purpose of this review is to describe the epidemiology, clinical features, putative immune mechanisms and management of immune reconstitution inflammatory syndrome (IRIS) using data published in the last 2 years. RECENT FINDINGS: Ever more conditions are reported as IRIS events. These most frequently occur with mycobacterial (tuberculosis or Mycobacterium avium complex infection) or cryptococcal disease (each in approximately 30% of cases). Definitions have been proposed for its clinical diagnosis. These suffer from a lack of prospective studies to support their predictive value. The immunopathogenesis of IRIS appears to be related to the interaction between HAART induced changes in host immune response and the presence of (usually microbial) antigen. Increasing evidence exists that this might be an anatomically compartmentalized phenomenon, such that immune responses may be localized to specific tissue sites such as the brain. This has implications for the use of simple blood tests, such as CD4 count or change in viral load, when assessing risk of IRIS. Treatment options include immune modulation, though supportive care is typically all that is required, unless symptoms are prolonged, significant or life-threatening. SUMMARY: IRIS is common and will become more so as HAART is rolled out worldwide. Clear clinical definitions are required to avoid its over diagnosis due to misclassification of other conditions. PMID- 16374214 TI - Changing patterns of sexual behaviour in the era of highly active antiretroviral therapy. AB - PURPOSE OF REVIEW: To describe changing patterns of sexual behaviour in the era of highly active antiretroviral therapy among gay/bisexual men in Europe, Canada, USA and Australia. RECENT FINDINGS: While there has been a substantial increase in high-risk sexual behaviour among gay men since 1996, this now appears to be levelling off in some cities. Overall the empirical evidence does not support the suggestion that taking highly active antiretroviral therapy or having an undetectable viral load leads to risky sexual behaviour among people with HIV. Nor can HIV treatment optimism alone explain the recent increase in high-risk sexual behaviour. Since 1996, an increasing number of gay men have begun to use the Internet to look for sexual partners. By serosorting on the Internet, HIV positive men are more likely to meet online, rather than off-line, other HIV positive men for unprotected sex. While serosorting does not present a risk of HIV transmission to an uninfected person, it does present a risk of other sexually transmitted infections and co-infection with resistant virus for HIV positive men themselves. This review also explores emerging behaviours such as barebacking and strategic positioning as well as the role of crystal meth and Viagra. SUMMARY: The review reminds us of the complexity of human and sexual behaviour. Among gay men, sexual behaviour in the era of highly active antiretroviral therapy has been characterized by risk reduction and stabilization as well as increasing risk. These changing patterns provide a new challenge as well as new opportunities for HIV prevention. PMID- 16374215 TI - Strategies to prevent mother-to-child transmission of HIV. AB - PURPOSE OF REVIEW: This review describes recent advances in the prevention of mother-to-child transmission, focusing on the use of antiretroviral treatment strategies in pregnancy, and discusses the emergence of viral resistance following the use of nevirapine to prevent mother-to-child transmission. RECENT FINDINGS: Mother-to-child transmission has been dramatically reduced in developed countries by the use of antiretroviral treatment and avoidance of breastfeeding. Highly active antiretroviral therapy use in pregnancy is recommended for women who require ongoing treatment, and, where available, is also very effective in reducing mother-to-child transmission in women with higher CD4 counts. The addition of a maternal and infant nevirapine dose to antenatal zidovudine can reduce transmission to below 5%, approximately half the transmission rate that can be achieved by single-dose nevirapine alone. The emergence of resistant virus following nevirapine use is a concern, occurring in up to 60% of mothers and 50% of infants following a single dose. Addition of zidovudine and lamivudine for 4-7 days postpartum can reduce the risk of resistance to 10%. SUMMARY: There is broad consensus on an approach to preventing mother-to-child transmission, which provides antiretroviral treatment in pregnancy and beyond to those women who need it, and an effective prophylactic regimen for those who do not yet need treatment, These regimens include highly active antiretroviral therapy, where available, a zidovudine-plus-nevirapine regimen in other settings, or nevirapine alone where this is all that is possible. More work is needed on the impact of nevirapine resistance and on reducing breast-milk transmission. PMID- 16374216 TI - Preventing sexually transmitted infections among adolescents: 'the glass is half full'. AB - PURPOSE OF REVIEW: Given the disproportionate burden of sexually transmitted infections for adolescents, there is an urgent need to identify effective prevention programs. RECENT FINDINGS: This review documents the efficacy of recent sexually transmitted infection-prevention programs. Overall, the review identified few sexually transmitted infection-prevention trials published since 2000. Moreover, considerable variability in program efficacy was observed across studies. Some studies observed changes in sexually transmitted infection associated risk behaviors, while only a few identified reductions in biologically confirmed sexually transmitted infections. In general, few programs demonstrated consistency of effects and a significant magnitude of effects across a broad range of outcomes. SUMMARY: New and innovative approaches are needed to amplify sexually transmitted infection intervention effects. Program development and evaluation needs to continue in a coordinated, scientifically rigorous fashion to optimize impact and, as important, to sustain effects over protracted periods. Furthermore, for interventions with demonstrated efficacy, a critical challenge is to translate them into sustainable programs that are widely disseminated. Ultimately, preventing sexually transmitted infections in adolescents does not only depend on the development of effective interventions alone, but on how effectively these interventions can be translated and integrated into self sustaining components of clinic, school or community programs, particularly in those areas and among adolescent populations most adversely impacted by the epidemic of sexually transmitted infection. PMID- 16374217 TI - Sexually transmitted diseases in incarcerated adolescents. AB - PURPOSE OF REVIEW: The objectives of this review are to summarize recent developments in the epidemiology of sexually transmitted disease in incarcerated adolescents and to review screening and treatment recommendations for sexually transmitted disease in juvenile corrections facilities. RECENT FINDINGS: The introduction of non-invasive, urine-based nucleic acid-amplification tests for chlamydia and gonorrhea has led to a dramatic increase in the ability to screen for chlamydia and gonorrhea in non-traditional settings, including corrections facilities. The prevalence of chlamydia and gonorrhea has been uniformly high in incarcerated adolescents. The prevalences of chlamydia and gonorrhea in adolescents aged 18-19 years incarcerated in adult corrections facilities were higher than those incarcerated in juvenile facilities. The prevalence was higher in incarcerated adolescent women than adolescent men and in black adolescents than non-black adolescents. Screening for chlamydia in incarcerated adolescents has been shown to be a cost-effective strategy for preventing adverse health consequences. Syphilis prevalence in incarcerated adolescents is relatively low. Hepatitis B is relatively common among incarcerated adolescents. SUMMARY: The high prevalence of chlamydia in incarcerated adolescents and gonorrhea in incarcerated adolescent women suggests that screening of these populations should be a priority. The reasons for the higher prevalence of chlamydia and gonorrhea in young adults aged 18-19 years incarcerated in selected adult corrections facilities compared with those incarcerated in juvenile facilities should be investigated. Hepatitis B virus vaccination in juvenile correction facilities is recommended to prevent and control the transmission of this disease. PMID- 16374218 TI - Prioritizing prevention of HIV and sexually transmitted infections: first generation vaginal microbicides. AB - PURPOSE OF REVIEW: As the HIV/AIDS pandemic continues unabated, novel control measures for the spread of HIV and other sexually transmitted infections are urgently needed. Topical microbicides are designed to prevent transmission of sexually transmitted infections when applied vaginally. The microbicides discussed in this review may provide a new opportunity for decreasing the spread of HIV and other sexually transmitted infections. RECENT FINDINGS: Epidemiological studies suggest a synergistic relationship between HIV and sexually transmitted infections, particularly between HIV and genital herpes infection. Compounds have been developed to block transmission of HIV-1 and herpes simplex virus, as well as Neisseria gonorrhoea and Chlamydia trachomatis. Several of these compounds have advanced to clinical trials as candidate microbicides. Candidate compounds fall into the following categories: detergents or surfactants that inactivate viral particles, anionic polymers that block attachment of virus to target cells, vaginal acid-buffering agents that maintain a protective vaginal pH, and antiretroviral drugs specific for HIV. Evaluation of the safety of topical microbicides remains problematic. Clinical experiences indicate that current models to assess safety in vitro and in vivo may be insufficient to assess the safety of vaginal microbicides. A critical direction of future studies is to identify which assay(s) provide surrogate laboratory markers of safety that correlate with clinical outcomes. SUMMARY: The spread of HIV, and its increasing burden of disease in women, necessitates the development of novel prophylactic strategies. Topical microbicides offer women an empowering preventative option but require vigorous testing for safety and effectiveness. PMID- 16374219 TI - Management of genital infections in pregnant women. AB - PURPOSE OF REVIEW: To examine and evaluate ways of managing genital infections in pregnant women. RECENT FINDINGS: The need to screen for sexually transmitted diseases during pregnancy depends on the prevalence of the condition, its pathogenesis and the cost-benefit analysis for a population or risk group. For a few genital infections with severe impact on the outcome of the pregnancy, such as syphilis and gonorrhoea, a 'screen and treat' policy is almost always cost effective. SUMMARY: Genital infections often remain unnoticed during pregnancy, as their signs and symptoms may be seen as part of the normal discomfort of pregnancy. Also it is sometimes not clear whether there are multiple partners or whether the partners have been treated, making re-infection after treatment highly likely. Partner tracing may be difficult, but the 'screen and treat' policy is usually the best solution. PMID- 16374220 TI - Human papillomavirus and anal intraepithelial neoplasia. AB - PURPOSE OF REVIEW: A review of recent developments in the understanding of the natural history of anal squamous carcinoma arising from areas of high-grade anal intraepithelial neoplasia. RECENT FINDINGS: Anal intraepithelial neoplasia is a consequence of chronic human papillomavirus infection in the anal canal and appears to be driven by high viral loads of human papillomavirus. In men who have sex with men with multiple sexual partners prevalent human papillomavirus infection does not decline with age, in contrast to heterosexual patients. Anal intraepithelial neoplasia is equally prevalent in different age groups of men who have sex with men, but in other respects what is known of its natural history resembles that of cervical intraepithelial neoplasia. Low-grade lesions frequently resolve, but high-grade lesions are much more stable. HIV-positives who practise receptive anal intercourse are at highest risk of anal intraepithelial neoplasia. Screening is easy to perform using cytology; the limitations of anal cytology being similar to those of cervical cytology. Patients with any grade of cytological abnormality require further investigation, ideally with high-resolution anoscopy, every 6 months. Successful treatments for individual small to medium-sized high-grade lesions include trichloroacetic acid, infra-red coagulation and laser. In HIV-positive patients the development of new lesions elsewhere is very likely. Topical agents for multifocal disease include imiquimod and cidofovir. SUMMARY: There is a need for large prospective cohort studies in men who have sex with men and HIV-positive patients to further our understanding of this disease and to evaluate treatment strategies. PMID- 16374221 TI - Epidemiology of intensive care unit-acquired urinary tract infections. AB - PURPOSE OF REVIEW: The development of urinary tract infections in critically ill adult patients is associated with considerable morbidity, prolonged hospitalization, and greater healthcare expenditures. We review the occurrence, microbiology, risk factors for acquisition, and outcomes associated with intensive care unit-acquired urinary tract infections. RECENT FINDINGS: Reports from several countries indicate that nosocomial urinary tract infections frequently complicate the course of patients admitted to intensive care units. Virtually all patients who develop an intensive care unit-acquired urinary tract infection have indwelling urinary catheters; other factors associated with the development of these infections include increased duration of urinary catheterization, female sex, intensive care unit length of stay, and preceding systemic antimicrobial therapy. The most frequent pathogens include Escherichia coli, Pseudomonas aeruginosa, enterococci, and Candida albicans; both the species distribution and rates of resistance vary considerably among institutions and regions. Secondary bloodstream infections are uncommon. Although acquisition of an intensive care unit-acquired urinary tract infection has been associated with a prolongation of intensive care unit length of stay, higher cost, and a higher crude case fatality rate, they do not appear to independently increase the risk for death. SUMMARY: Urinary tract infection is a common complication of critical illness that is associated with increased patient morbidity but not mortality. There is a relative paucity of research on nosocomial urinary tract infection specifically acquired in the intensive care unit and further studies are needed to better define the epidemiology and management of these infections. PMID- 16374223 TI - Trends in cancer mortality in Switzerland, 1980-2001. AB - Trends in cancer mortality in Switzerland were analysed over the period 1980 2001, on the basis of the World Health Organization database. Appropriately developed correction factors were utilized for the period before 1995, to allow for spurious trends introduced by the change between the 8th and the 10th revisions of the ICD. Steady declines in cancer mortality were observed, particularly from the mid-1980s onwards. Over the last decade, the fall in overall age-standardized (world standard) cancer mortality was 11.1% in men (from 158.1 in 1990-1991 to 140.6/100,000 in 2000-2001) and 7.6% in women (from 91.6 to 84.7/100,000), and the decline was larger in truncated rates from 35 to 64 years (-18.0 and -9.7%). In men, all major tobacco and alcohol neoplasms have declined until the late 1990s but have levelled off over the last few years, reflecting recent trends in alcohol and tobacco consumption. The fall in male lung cancer mortality was 20% over the last decade (from 42.9 to 34.3/100,000). In contrast, lung cancer mortality in women has steadily increased by 38% between 1981 and 1991 and by 47% between 1991 and 2001, to reach 10.7/100,000 at all ages and 18.3 at age 35 to 64, due to increased prevalence of smoking in subsequent generations of Swiss women. Other sites showing substantial declines include stomach and colorectum in both sexes, (cervix) uteri and breast in women. Likewise, prostate cancer showed modest favourable trends after 1995. Steady declines were observed for leukaemias, Hodgkin's disease and testicular cancer, namely, the neoplasms most influenced by therapeutic improvements, while trends in lymphomas and myeloma showed no clear pattern. PMID- 16374224 TI - Lifestyle characteristics among participants in a Norwegian colorectal cancer screening trial. AB - The aim of the present study was to evaluate to what extent lifestyle-related variables predict participation for flexible sigmoidoscopy (FS) screening when the compliance is relatively high. During 2001, a randomized sample of 6961 men and women, born between 1946 and 1950, were invited to have a flexible sigmoidoscopy screening examination. Attendees (n = 4111) were asked to fill in a questionnaire focusing on physical activity, body weight, smoking habits and diet. The questionnaire was sent by mail to non-attendees (n = 2628) and a randomized corresponding control group not invited to screening (n = 7000). Sixty one percent attended for screening. The questionnaire response rate among attendees, non-attendees and controls was 97, 11 and 61%, respectively. Attendees were more physically active (P < 0.001), and showed more adherence to general dietary recommendations, compared with controls. Opposing their healthy exercise and dietary habits, however, attendees were more likely to be moderate smokers compared with controls. The present offer of FS screening, achieving a relatively high compliance rate, may have reached attendees using screening as a supplement to an already healthy lifestyle, but also those who seek amelioration through health checks for risks behaviour that they are perfectly well aware of, such as smoking. PMID- 16374225 TI - Diet during pregnancy and levels of maternal pregnancy hormones in relation to the risk of breast cancer in the offspring. AB - Birth weight is positively associated with the risk of breast cancer in the offspring and the underlying process is likely to involve the pregnancy endocrine milieu. We have examined the association of diet and related factors during pregnancy with the levels (at the 16th and 27th gestational week) of maternal pregnancy oestradiol, oestriol, sex hormone-binding globulin (SHBG), progesterone and prolactin, in a cohort of 270 Caucasian women who delivered in a major hospital in Boston, USA. Oestradiol and oestriol were not strongly associated with any of the diet-related variables, but SHBG was significantly and consistently related inversely to pre-pregnancy body mass index and weight gain during pregnancy, and positively to vegetable and pulses intake. Pregnancy progesterone was associated positively with alcohol and inversely with polyunsaturated lipid and vitamin B12 intake, whereas pregnancy prolactin was inversely associated with cereal consumption. If the pregnancy hormones studied are indeed involved in the intra-uterine origin of breast cancer, these findings, if confirmed, would focus dietary advice to pregnant women, with a view to reducing the risk of breast cancer in the offspring, towards avoidance of excess energy intake and an emphasis on plant foods. This advice does not contradict current dietary advice on prudent diet during pregnancy and throughout life. PMID- 16374226 TI - Determinants of non-compliance to recommendations on breast cancer screening among women participating in the French E3N cohort study. AB - Evidence of the benefits of screening for breast cancer using mammography after the age of 50 is considered sufficient. Information on the determinants of compliance to the recommendations on mammography is needed to identify women to which public health messages should be specifically addressed and also to interpret results from epidemiological data in which some breast cancer cases are detected through screening programmes and others are not. The general characteristics and dietary data of French women participating in the E3N cohort study were analysed. Odds ratios of the frequency of non-compliance to recommendations on breast cancer screening were computed in women over and under the age of 50. Non-compliant women over 50 (i.e. never attenders) had a poorer access to physicians and poorer health with regard to specific risk factors even after adjustment for age and educational level. Women who were used to referring to gynaecologists early in life were better attenders later on. Logically, women under 50 who were over-screened, as compared to public health recommendations, had the opposite characteristics. Public health recommendations should be designed specifically according to targeted subgroups of women. Determinants of attendance for screening should be kept in mind in the interpretation of epidemiological studies in which some cases may be over-screened and others not. PMID- 16374227 TI - Performance measures from 10 years of breast screening in the Ontario Breast Screening Program, 1990/91 to 2000. AB - Performance measures for the Ontario Breast Screening Program (OBSP) by age group, time period and screening modality from 10 years of breast screening were evaluated. Data were available from routine information collected on 283,962 women aged 50 to 69 screened at 73 screening centres between 1 July 1990 and 31 December 2000. Although, initially, participation in the OBSP was low, this rate increased over time and the majority of women screened returned for subsequent screening. Abnormal call rates increased slightly over the time period, were higher in women aged 50 to 59, and for women with mammographic abnormalities. Detection rates of invasive cancer were higher and prognostic features of cancers were better for women age 60 to 69, and those referred by mammography. Along with the prognostic features of cancers, the benign to malignant surgical ratio and diagnostic interval improved over the time periods and for women aged 60 to 69. Greater proportions of women had shorter diagnostic intervals and were more likely to have a diagnosis of breast cancer after surgery if they were referred by both clinical breast examination and mammography. Although some enhancements of the programme are necessary, the OBSP met or exceeded Canadian targets for most performance measures. PMID- 16374228 TI - Aspirin and the risk of prostate cancer. AB - Epidemiological studies have suggested that aspirin may have a protective effect against prostate cancer, but the evidence is still limited and inconsistent. The role of aspirin in prostate cancer risk was analysed in a multicentric case control study conducted in Italy between 1991 and 2002, including 1261 incident cases of carcinoma of the prostate and 1131 hospital controls. A total of 115 (9.1%) cases versus 90 (8.0%) controls reported regular aspirin use. The multivariate odds ratio (OR) for regular aspirin users was 1.10 (95% confidence interval (95% CI) 0.81-1.50). No relation was found with duration of use (OR = 1.03 for <5 years, and 1.17 for > or =5 years) and time since first use (OR = 1.03 for <10 years, and 1.35 for > or =10 years). These findings do not support a protective role of regular aspirin use on prostate cancer risk. PMID- 16374229 TI - Polymorphisms of methylenetetrahydrofolate reductase and the risk of prostate cancer: a nested case-control study. AB - It has been proposed that folate and polymorphisms of the enzyme methylenetetrahydrofolate reductase (MTHFR), which regulates influx of folate from DNA synthesis and repair to methylation reactions, are involved in the aetiology of cancer. To relate the MTHFR 677C-->T and 1298A-->C polymorphisms to the risk of prostate cancer, taking into consideration prospective plasma levels of folate, vitamin B12 and homocysteine. The design was a case-control study of 223 prostate cancer cases and 435 matched controls nested within the population based Northern Sweden Health and Disease Cohort. Neither the MTHFR 677C-->T nor the MTHFR 1298A-->C polymorphism was statistically significantly associated with the risk of prostate cancer in univariate analysis by conditional logistic regression. After adjustment for MTHFR 1298A-->C, plasma folate, vitamin B12, homocysteine, body mass index and smoking, the odds ratios were, for the 677 CT genotype, 1.52 [95% confidence interval (CI) 1.02-2.26], and for TT, 0.91 (95% CI 0.41-2.04). Our previously reported observation of a possible increase in the risk of prostate cancer at high plasma folate levels was attributable in this study to subjects having the MTHFR 677C-->T polymorphism. We found that the MTHFR 677C-->T polymorphism is not likely to have a major role in the development of prostate cancer, although it may possibly increase the risk in combination with high plasma folate levels. Further investigation in larger studies is warranted. PMID- 16374230 TI - Trends and patterns in prostate cancer mortality in Belgrade, Serbia: a join point analysis. AB - The objective of this analysis was to examine recent changes in prostate cancer mortality among the male population of Belgrade, Serbia, during the period 1975 2002. Mortality data (official death certificates) for prostate cancer were gathered from published and unpublished material of the Municipal Institute of Statistics in Belgrade. Mortality rates were standardized according to the European standard population. Trends in mortality rates were assessed using join point analysis. During the period from 1975 to 2002 in the population of Belgrade the average age-adjusted mortality rate from prostate cancer during this period was 17.5/100,000 (95% confidence interval, 15.4-19.6), ranging from 8.2/100,000 in 1984 to 31.2/100,000 in 2002. Since 1986 mortality rates among men aged 50 years and over have risen by nearly 6% per year, with the increases being slightly higher among older men (70 years and over 8% per year) compared to younger men (50-69 years, 4% per year). Mortality due to prostate cancer is rising quickly among men in Belgrade. This level of increase is the highest of any reported increase internationally and contrasts sharply with the widespread decrease in prostate cancer mortality in many developed countries. PMID- 16374231 TI - Critical role of the c-JunNH2-terminal kinase and p38 mitogen-activated protein kinase pathways on sodium butyrate-induced apoptosis in DU145 human prostate cancer cells. AB - Sodium butyrate (NaBu) is known to exhibit anti-cancer effects via the differentiation and apoptosis of various carcinoma cells. However, the mechanism by which NaBu induces apoptosis and the involvement of protein kinases during apoptosis is not completely understood. To investigate the underlying pathways, we performed cell culture experiments in androgen-independent human prostate cancer (DU145 cells) focusing on various protein kinases. NaBu causes concentration-dependent cell detachment and growth inhibition. Exposure of DU145 cells to NaBu for 24 h caused a strong apoptotic effect with 26% nuclear fragmentation and condensation. In addition, NaBu induced caspase-3 and poly-ADP ribose polymerase cleavage and up-regulation of bax, suggesting that mitochondrial damage is involved in NaBu-induced caspase-dependent apoptosis. Interestingly, NaBu stimulated p38 mitogen-activated protein kinase (MAPK) and c Jun NH2-terminal kinase (JNK) activation, but not extracellular signal-regulated kinase 1/2 activation during apoptosis. Furthermore, NaBu up-regulated total protein levels and phospho forms of MAPK kinase 3 (MKK3) and MAPK kinase 4 (MKK4) as the upstream kinases of p38 MAPK and JNK independently of oxidative stress. Taken together, it is suggested that NaBu can be a promising chemopreventive agent for prostate cancer and the p38 MAPK and JNK pathways have critical roles in NaBu-induced apoptosis in DU145 cells. PMID- 16374232 TI - Clinicopathological features of melanocytic skin lesions in Egypt. AB - Although melanocytic skin lesions have been recognized since antiquity, their literature was limited to Caucasians. To date, the clinicopathologic features of these lesions in Egyptians are still unknown. To define these features, diagnostic records of the melanocytic skin lesions received at the Pathology Department, Assuit University Hospitals (1989-2004) were reviewed. The lesions examined included 12 benign naevi (BN), 10 dysplastic naevi (DN), and 21 cutaneous malignant melanomas (CMMs). The DN and CMMs were more common in men than in women (2 : 1 and 1.5 : 1, respectively) while BN were more common in women (2 : 1). The average age incidence was 33+/-5, 38+/-7 and 54+/-3 years, for BN, DN and CMM, respectively. The lower limb (13/21, 62%), head and neck (7/21, 33%) were the most common sites for CMMs. The average size (mm) was 2+/-0.3, 4+/ 0.6 and 21+/-0.3 for BN, DN and CMMs, respectively. Recurrence occurred in 10% of CMMs. Histologically, CMMs were of nodular type and composed of epithelioid (7/21, 33%), spindle cells (1/21, 5%), or mixed cells (13/21, 62%). The mean tumour thickness (Breslow) was 6+/-0.5 mm. CMMs included two of 21(9%), three of 21(14%), six of 21(38%), and 10 of 21(38%) with Clark level II, III, IV and V. In Egypt, CMM is the third most common cutaneous neoplasm following squamous and basal cell carcinomas. Compared with Western societies, melanoma has a male sex predilection, similar histological features but different topographical distribution and rare incidence. The striking difference from Western series is the incidence of nodular melanoma - in the West this represents 15-30% of melanomas, with superficial spreading being the majority. Another key difference from the West is the 'sun-bed' culture of the West and the desire to have suntans. This is the first study that reports the clinicopathologic features of melanocytic skin lesions in Egypt. PMID- 16374234 TI - Pizza consumption and the risk of breast, ovarian and prostate cancer. AB - Pizza has been favourably related to the risk of prostate cancer in North America. Scanty information, however, is available on sex hormone-related cancer sites. We therefore studied the role of pizza consumption on the risk of breast, ovarian and prostate cancers using data from three hospital-based case-control studies conducted in Italy between 1991 and 2002. These included 2569 women with breast cancer, 1031 with ovarian cancer, 1294 men with prostate cancer, and a total of 4864 controls. Compared with non-pizza eaters, the multivariate odds ratios for eaters were 0.97 (95% confidence interval (CI) 0.86-1.10) for breast, 1.06 (95% CI 0.89-1.26) for ovarian and 1.04 (95% CI 0.88-1.23) for prostate cancer. Corresponding estimates for regular eaters (i.e. > or =1 portion per week) were 0.92 (95% CI 0.78-1.08), 1.00 (95% CI 0.80-1.25) and 1.12 (95% CI 0.88 1.43), respectively. Our results do not show a relevant role of pizza on the risk of sex hormone-related cancers. The difference with selected studies from North America suggests that dietary and lifestyle correlates of pizza eating vary between different populations and social groups. PMID- 16374233 TI - Type of alcoholic beverage and the risk of laryngeal cancer. AB - A few studies have investigated whether the risk of laryngeal cancer depends on the types of alcoholic beverage consumed, providing conflicting results. We investigated this issue using the data from two case-control studies conducted in Italy between 1986 and 2000. These included 672 cases of laryngeal cancer and 3454 hospital controls, admitted for acute, non-neoplastic conditions, unrelated to smoking and alcohol consumption. Significant trends in risk were found for total alcohol intake, with multivariate odds ratios (ORs) of 1.12 for drinkers of 3-4 drinks/day, 2.43 for 5-7, 3.65 for 8-11, and 4.83 for > 12 drinks/day, as compared to abstainers or light drinkers. Corresponding ORs for wine drinkers were 1.12, 2.45, 3.29 and 5.91. After allowance was made for wine intake, the ORs for beer drinkers were 1.65 for 1-2 drinks/day, and 1.36 for > or = 3 drinks/day, as compared to non-beer drinkers; corresponding values for spirits drinkers were 0.88 and 1.15. This study thus indicates that in the Italian population characterized by frequent wine consumption, wine is the beverage most strongly related to the risk of laryngeal cancer. PMID- 16374235 TI - Prevalence of smoking and attitude towards smoking regulation in Italy, 2004. AB - AIMS AND BACKGROUND: To update trends in smoking prevalence in Italy to 2004, and describe the attitudes towards forthcoming regulation of smoking. METHODS: Population-based face-to-face survey conducted in March-April 2004 on a sample of 3050 individuals (1509 men and 1541 women) aged 15 or over, representative of the general adult Italian population in terms of age, sex, geographic area, habitat, education and working status. RESULTS: Overall, 26.2% of the Italians aged 15 or over described themselves as current cigarette smokers (30.0% of men, 22.5% of women); 19.1% of men and 11.2% of women smoked 15 or more cigarettes per day. Ex smokers were 17.9% (24.8% of men, 11.2% of women). The difference in smoking prevalence between men and women was greater in the elderly. More educated men, and those residing in northern Italy were less frequently current smokers. Conversely, women with a low level of education (19%) were less frequently current smokers than those with higher education (24%). More than 85% of Italian adult population were in favour of restrictions of smoking in public places, such as cafes and restaurants, and to ban smoking in workplaces. CONCLUSIONS: Self reported smoking prevalence is decreasing in men but not in females. There is widespread support for smoking restriction in public areas. PMID- 16374236 TI - Intention to adopt a smoking cessation expert system within a self-selected sample of Dutch general practitioners. AB - To investigate intention to adopt a new smoking cessation expert system as well as outline perceived barriers by general practitioners (GPs) to adopt this expert system, a written questionnaire was sent to 771 registered GPs. Respondents, representing 34.8% of the registered GPs, were classified as adopters (34.2%), doubters (36.2%) or non-adopters (29.2%). Adopters and doubters were less negative about the time investment for the GP when adopting the expert system than non-adopters. Adopters expected a more positive reaction from their patients than non-adopters. Smoking cessation was mostly considered to be a task for the practice assistant. The authors discuss the relevance of barriers mentioned not to implement the expert system and give recommendations for further steps into implementing primary prevention activities in Dutch general practice. PMID- 16374237 TI - Does smoking influence survival in cancer patients through effects on respiratory and vascular disease? AB - Patients with cancers caused by smoking may die because they continue to smoke even after diagnosis of a cancer caused by smoking. We investigated differences in cause-specific mortality between patients diagnosed with smoking-related and non-smoking-related cancers. The causes of death were classified as smoking related cancer, non-smoking-related cancer, respiratory or vascular disease, and all other causes. We studied all 220 089 people diagnosed with cancer in Scotland between 1986 and 1996, aged between 20 and 85 years, with last follow-up on 31 December 1999. There was a moderate excess risk of dying from respiratory and vascular causes in those with smoking-related cancers, which did not fall with time since diagnosis, consistent with continued smoking by these patients. Mortality among cancer patients might fall if more assistance in stopping smoking was provided for patients who have smoking-related cancers. PMID- 16374238 TI - Oncologists' opinions about research ethics committees in Italy: an update, 2004. AB - While there is general consensus about the basic features of ethical research, the explosion of information from technology, genetics and clinical research, and the desire to accelerate the translation of preliminary findings into clinical research and practice have created space for conflicts between the aims of research, medical ethics and human rights. Research ethics committees (RECs) play a crucial role. In 2004, a short standardized questionnaire was sent to 110 Italian oncologists. The questionnaire explored aspects of the interaction between oncologists and RECs, including reports on personal experience with RECs, and the oncologists' opinions and level of satisfaction with important functions of RECs, such as bureaucratic, ethics, scientific, and educational and training aspects. The findings are compared with those from the previous surveys. Eighty three out of the 91 oncologists returned the questionnaire. Responders were working in 67 hospitals (60 RECs). The level of satisfaction with REC activities was high for ethical and scientific aspects (87 and 80% were satisfied or very satisfied, respectively); somewhat lower for bureaucratic and secretarial aspects (61%); and much lower for educational and formation activities (33%). Comparison with other surveys does not show any important difference. The main findings of this survey are the stable figures concerning oncologists' opinions and the low ratings given to education and training aspects. This latter finding clearly identifies the need for educational activities, and the possibility of using this approach to foster interactions between RECs and researchers. PMID- 16374239 TI - First aid, fast track and the fertile fields of peer review. PMID- 16374240 TI - Diagnostic accuracy of lower extremity X-ray interpretation by 'specialized' emergency nurses. AB - OBJECTIVES: In the quest for a cost-effective and quality-preserving solution to manage crowding in the emergency department, the possibility of deploying regular emergency nurses for the treatment of acute ankle injuries was investigated. The aim of this study is to compare the diagnostic accuracy of emergency nurses with that of senior house officers in interpreting ankle and foot radiographs. METHODS: A prospective study comparing the assessment of 60 radiographs (30 feet and 30 ankles) by 16 emergency nurses before and after an educational session was performed. Each subset of 30 radiographs contained 12 fractures, hand-picked by a radiologist to represent everyday traumatology in the emergency department. The control group consisted of eight senior house officers representing everyday expertise. The outcome of the diagnostic assessment, represented as the pooled sensitivity and specificity for both groups, was compared using Z-statistics. RESULTS: Before the training session, the specialized emergency nurse group showed a sensitivity of 0.87 (confidence interval 0.83-0.91) compared with 0.93 (confidence interval 0.88-0.96) for the control group (P = 0.05). The specificity of specialized emergency nurses was 0.87 (confidence interval 0.81-0.92) compared with 0.93 (confidence interval 0.89-0.95) for the senior house officers (P < 0.05). After the training session, specialized emergency nurse diagnostic parameters did not differ significantly from the control group, displaying a sensitivity of 0.89 (confidence interval 0.86-0.92) and specificity of 0.92 (confidence interval 0.87-0.95). CONCLUSION: Before the training session, the specialized emergency nurse group showed a significantly lower accuracy than the SHO group. After training, however, the diagnostic accuracy did not differ significantly between groups. Therefore, we conclude that emergency nurses are able to accurately interpret foot and ankle radiographs after a short educational session. PMID- 16374241 TI - Training of 'first-aid' trainers: a medical school example in Turkey. AB - OBJECTIVE: A student-oriented basic life support and first-aid training is practical to enlarge the number of qualified people in the field. In this study, we aimed to describe and discuss the training process of a group of medical students at Hacettepe University, School of Medicine and their assessment as basic life support and first-aid trainers. METHODS: A '6-hour' 'Training the Trainers' course was conducted and participants' demonstration and coaching skills were evaluated by using a 3-point scale (1 - needs improvement; 2 - competently performed; 3 - proficiently performed). In total, 31 students out of 45 volunteers were certified as trainers. RESULTS: All of the students proficiently performed 'explain the aim of the demonstration', 'motivate participants to use checklists', and 'motivate the participants to ask questions' for demonstraton skills. Similarly, they all got the maximum grade for 'answer the questions of the participant', 'discuss the coach role of the participant', 'apply the skills', and 'follow up with the participant by using the checklist' as steps of coaching skills. None of the skills within the checklists needed improvement for any of the candidates. CONCLUSION: Although there are limitations, we hope that our experience of the Training the Trainers course will be useful for readers to highlight the importance of skill-based peer education on a very important public health concern, especially for the developing countries. PMID- 16374242 TI - Out-of-hospital resuscitation in Estonia: a bystander-witnessed sudden cardiac arrest. AB - OBJECTIVE: To evaluate the results of the first epidemiological study on out-of hospital resuscitation in Estonia. METHODS: A prospective cohort study of 2108 consecutive standardized reports on out-of-hospital resuscitation attempts from 1 January 1999 to 31 December 2002 was conducted according to the Utstein style. RESULTS: In all, 67.3% (1419/2108) of the cardiac arrests were of presumed cardiac aetiology and 60.2% (854/1419) of them were bystander-witnessed. Of these, the 28% bystander cardiopulmonary resuscitation was initiated, and the first rhythm was recorded as ventricular fibrillation or pulseless ventricular tachycardia in 40% of the cases. In the subgroup of patients with bystander witnessed cardiac arrest of cardiac origin, 10.7% (91/854) were discharged alive in good cerebral performance categories and 7.7% were alive at the 1-year follow up. The chances of survival increased if the median response time interval was <6 min, cardiac arrest occurred in a public place, patients received bystander cardiopulmonary resuscitation and had an initial rhythm of ventricular fibrillation or pulseless ventricular tachycardia. The discharge rate was 24% (82/343) in the subgroup of patients who had bystander-witnessed cardiac arrest of cardiac origin and an initial rhythm of ventricular fibrillation or pulseless ventricular tachycardia. In this subgroup, the survival rate was 42.6% (40/94) in Tartu urban area, 16.9% (22/130) in Tallinn urban area and 16.8% (20/119) in other regions of Estonia (mostly urban and suburban areas). CONCLUSION: The results demonstrate that despite the progress in the management of out-of hospital cardiac arrest in Estonia, only one centre (Tartu) achieves a better survival rate. Further improvements are needed to raise the quality of the Estonian emergency medical services system, especially in rural areas. PMID- 16374243 TI - Chronic outpatient management of asthmatics attending the emergency department: a survey from a country with low income. AB - PURPOSE: Little is known about compliance with international guidelines of asthma management in developing countries where some medications are prohibitively expensive. METHODS: A survey was conducted in asthmatic patients attending the emergency department for acute asthma. Asthma severity was evaluated and conformity of chronic treatment with international guidelines was assessed. Additional features of asthmatic education were also evaluated. RESULTS: A total of 127 consecutive patients (mean age 34 +/- 14 years) answered the questionnaire. Mild asthma was present in 19.7% patients, 56.7% had moderate asthma and 23.6% had severe asthma. Of the 124 known asthmatic patients, 33% had no treatment for chronic asthma. In the remaining, treatment adhered to international guidelines in 44% patients. The major cause of treatment inadequacy was the lack of inhaled corticosteroids (64%) or suboptimal dosage of corticosteroids (13%). Conformity to guidelines according to favorable or unfavorable economic conditions was 59% and 33%, respectively (P = 0.036). Treating physicians provided an 'action plan' for managing acute symptoms to 19% patients. Forty percent of asthmatic patients performed correctly the five components of metered dose inhaler use. CONCLUSION: Our study reveals an important proportion of non-treated asthmatic patients. In most asthmatic patients, treatment did not conform with guidelines because of an underutilization of corticosteroids, mainly because of economic obstacles. PMID- 16374244 TI - Paracetamol and salicylate testing: routinely required for all overdose patients? AB - OBJECTIVES: To determine whether it is necessary to routinely measure serum salicylate levels in addition to serum paracetamol levels in alert overdose patients who deny ingestion of that substance. METHODS: Prospective observational study of overdose patients attending an urban emergency department. RESULTS: A total of 171 patients attended with an alleged overdose. Thirty-seven patients were excluded; 24 patients had a reduced conscious level (Glasgow Coma Scale <15), nine patients could not recall the time of overdose and four patients had a staggered overdose. In all, 47.0% (63/134) claimed to have taken paracetamol and 5.2% (7/134) claimed to have taken salicylate. No patient who denied taking paracetamol or salicylate subsequently tested positive for them. In all, 51.5% (69/134) of patients reported that they had taken alcohol at the time of their overdose. CONCLUSION: Given the relatively low clinical risk and evident symptoms and signs from salicylate overdose, routine testing may not be required for fully conscious asymptomatic patients who deny taking it. The risks of delayed toxicity and lack of initial symptoms in those ingesting paracetamol suggests that continued routine testing is necessary. PMID- 16374245 TI - Procedure lab used to improve confidence in the performance of rarely performed procedures. AB - OBJECTIVES: The objective of this study is to assess the efficacy of an animal procedure lab in improving the level of comfort in performing important emergency medicine procedures. The procedures included central venous line, chest tube, cricothyrotomy, pericardiocentesis, venous cutdown, and thoracotomy. METHODS: The students were physicians participating in the Tuscan Emergency Medicine Initiative as part of a certificate program in emergency medicine. They attended a 1-h lecture discussing the procedures to be performed. Participants filled out a questionnaire before and after the lab, which asked how many times they had performed each procedure, how comfortable they felt, on a five-point scale, performing each procedure, and whether they felt comfortable performing it by themselves, with assistance or whether they would not feel comfortable performing the procedure. Differences in rated numerical values for each procedure before and after the lab were analyzed using a two-tailed t-test. Alpha was set at 0.95. RESULTS: In all, there were 20 participants with a wide range of experience. A statistical improvement was observed in comfort level and willingness to perform the procedures independently (P<0.01). The only non-significant change was in willingness to perform central lines. CONCLUSIONS: The use of an animal lab improves the comfort level of practitioners in performing procedures. Although procedures are best learned on patients with supervision, this is not always feasible. This lab is a useful adjunct to teaching in emergency medicine and allows participants exposure to critical procedures. PMID- 16374246 TI - How does fast track affect quality of care in the emergency department? AB - STUDY OBJECTIVES: Use of fast track has been shown to improve the emergency department flow of less urgent patients. It has been speculated, however, that this could negatively affect the care of urgent patients. The objective of this study was to determine whether a dedicated fast track for less urgent patients [Canadian Triage and Acuity scale category 4/5 (CTAS 4/5)] affected (1) the time to assessment for urgent patients (CTAS 3), (2) the length of stay for less urgent patients (CTAS 4 and 5), and (3) the left-without-being-seen rate. METHODS: In June 2003, fast track was opened in our emergency department from 13:00 to 19:00 h. A before-after intervention comparison analysis was completed for 1 week in Aug 2002 and the same week in Aug 2003. Data collected included (1) time to assessment of CTAS 3 patients, (2) the length of stay for CTAS 4/5 patients, and (3) percentage of patients who left without being seen. RESULTS: A total of 368 patients were reviewed for 2002 and 380 patients were reviewed for 2003. Median time to assessment of CTAS 3 patients presenting from 13:00 to 19:00 h was reduced from 66 min (Interquartile range: 40, 94 min) in 2002 to 60 min (IQR: 38, 108 min) after fast track was open in 2003 (P = 0.95). Median length of stay of CTAS 4 and 5 patients was reduced from 170 min (IQR: 111, 256 min) to 110 min (IQR: 69, 185 min) (P < 0.001). The overall left-without-being-seen rate decreased from 5% (20/368) to 2% (9/380). CONCLUSION: A dedicated fast track for CTAS 4/5 patients can reduce the length of stay and the left-without-being-seen rate with no impact on CTAS 3 patients seen in the main emergency department. PMID- 16374247 TI - Acute spontaneous cervical spinal epidural hematoma with hemiparesis as the initial presentation. AB - Acute onset of hemiparesis is a common initial presentation of cerebral vascular accidents, but many differential diagnoses should be considered. Hemiparesis results from an upper motor neuron lesion above the midcervical spinal cord, and the absence of cranial nerve signs or facial palsy suggests a lesion in the high cervical spinal cord. Most spinal cord lesions, however, result in quadriparesis or paraparesis, but not hemiparesis. We present a rare case of acute spontaneous cervical spinal epidural hematoma, with initial presentation of right hemiparesis and mild neck pain. PMID- 16374248 TI - Reversible visual loss in a patient with rheumatoid arthritis and the role of vascular imaging. AB - Spontaneous vertebral artery dissection is a condition that can have lethal consequences. The condition should be considered in young male patients who present with a stroke. At presentation, headaches, cerebral ischaemic episodes and oculosympathetic paresis are the most commonly encountered manifestations. The diagnosis is confirmed with angiography. Here, we present a middle-aged male gardener with rheumatoid arthritis and signs of vertebral artery dissection to highlight the importance of diagnosis and discuss the controversies in management. PMID- 16374249 TI - Paroxysmal sympathetic surge associated with gamma hydroxybutyrate. AB - Gamma hydroxybutyrate is increasingly being used recreationally in the United Kingdom. We present a case of gamma hydroxybutyrate overdose associated with paroxysmal sympathetic storm, a phenomenon usually confined to patients who have sustained traumatic brain injury. PMID- 16374250 TI - Treatment of acute myocardial infarction in Scottish emergency departments: survey of current practice. AB - INTRODUCTION: Thrombolysis was rarely given in emergency departments in Scotland when last studied in 1996. This study aimed to review the current practice of Scottish emergency departments with respect to thrombolysis for acute myocardial infarction. METHODS: Postal questionnaires were sent to all emergency departments in Scotland staffed by at least one consultant in emergency medicine, assessing the processes used for thrombolysis of acute myocardial infarction in the emergency department. A reminder letter was sent to non-responders after 1 month. All results were anonymized. RESULTS: The response rate was 77% (23 questionnaires from 30 hospitals). Twenty (87%) emergency departments performed thrombolysis according to protocol. In 13 emergency departments, thrombolysis was initiated by the emergency department staff, in six by on-call physicians and one emergency department had nurse-led thrombolysis. Twelve emergency departments occasionally received patients who had been given prehospital thrombolysis. Six hospitals had on-site primary angioplasty but only two hospitals had a 24-h service. Thirteen respondents thought the emergency department was the most appropriate place for thrombolysis, four felt that prehospital thrombolysis was best and one thought that coronary care was optimal. Four respondents felt that prehospital or the emergency department were the best options. CONCLUSION: Most emergency departments in Scotland are now administering thrombolysis for patients with acute myocardial infarction. Thrombolysis, delivered either in the prehospital arena or in the emergency department, is likely to be the primary option for patients with acute myocardial infarction in Scotland in the foreseeable future. PMID- 16374251 TI - Toll-like receptor-1, -2, and -6 polymorphisms influence disease extension in inflammatory bowel diseases. AB - BACKGROUND: Evidence that a deficient innate immune response toward the bacterial flora of the gut plays a role in the pathogenesis of inflammatory bowel disease (IBD) is growing. This is underscored by the finding of the association between CARD15 variants and Crohn's disease (CD) and D299G in Toll-like receptor (TLR) 4 and IBD. Our aims were to study nonsynonymous polymorphisms in other TLR genes in IBD. METHODS: Thirty-five single nucleotide polymorphisms (SNP) in TLR1-10 were identified from public databases. 284 IBD parent-child trios and a second independent cohort of 285 IBD patients and 191 healthy controls were genotyped with polymerase chain reaction-restriction fragment length polymorphisms. Patients were pooled for genotype-phenotype analyses. RESULTS: Although none of the SNPs was involved in disease susceptibility, a number of variants influenced the disease phenotype. A positive association between TLR1 R80T and pancolitis in UC (P = .045, OR [95% CI] 2.844 [1.026-7.844]) was found. The TLR2 R753G SNP was also associated with pancolitis (P = .027, OR [95% CI] 4.741 [1.197-18.773]). The relative risks for heterozygous patients to develop pancolitis were 5.8 and 3.3 for R80T and R753G, respectively. There was a negative association between TLR6 S249P and ulcerative colitis with proctitis only (P = .026, OR [95% CI] 0.223 [0.096-0.705]). In CD, we found a negative association between ileal disease involvement and TLR1 S602I (P = .03, OR [95% CI] 0.522 [0.286-0.950]). CONCLUSION: TLR2 and its cofactors TLR1 and TLR6 are involved in the initial immune response to bacteria by recognizing peptidoglycan. An association between nonsynonymous variants in the TLR1, -2, and -6 genes and extensive colonic disease in UC and CD was found. Our findings further highlight the role of an abnormal innate immune response in the pathogenesis of IBD. PMID- 16374253 TI - Increased expression of CD146, a new marker of the endothelial junction in active inflammatory bowel disease. AB - BACKGROUND: Crohn's disease (CD) and ulcerative colitis (UC), the 2 major forms of inflammatory bowel diseases (IBD), have been associated with disturbances in vascular physiology, including permeability and angiogenesis, that are in part regulated by the endothelial intercellular junctions. These junctions are composed of several adhesion molecules including the platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) and the more recently described CD146 (S Endo1 Ag, MUC18). AIM: To study the expression of tissue and soluble form of CD146 in patients with CD or UC in relation to disease activity and location. This study was made in comparison with the soluble form of CD31 (sCD31). RESULTS: In active disease, a high expression of CD146 was observed on endothelial cells in intestinal biopsies from both CD and UC. In addition, we observed a decrease of sCD146 in relation to active disease and extensive location of CD and UC. Lower levels of sCD31 were also detected in active and extensive location of UC, but no difference could be observed in CD. CONCLUSION: sCD146 is a novel marker of the endothelial intercellular junction that reflects endothelial remodeling more effectively than soluble CD31. Further studies are warranted to determine whether sCD146 will provide a serological assay reflecting alterations in vascular permeability and vessel proliferation in the inflamed IBD intestine. PMID- 16374252 TI - Both IL-12p70 and IL-23 are synthesized during active Crohn's disease and are down-regulated by treatment with anti-IL-12 p40 monoclonal antibody. AB - BACKGROUND: Interleukin (IL)-12p70 and IL-23 are key T helper-1 (TH1) cytokines that drive the inflammation seen in numerous models of intestinal inflammation. These molecules contain an identical p40 chain that is bound to a p35 chain in IL 12 and a p19 chain in IL-23, making both potentially susceptible to modulation by an anti-IL-12p40 monoclonal antibody (mAb). METHODS: In the present study, we sought to determine whether active inflammation in Crohn's disease (CD) is associated with the increased synthesis of both of these cytokines and whether patients treated with an anti-IL-12p40 mAb down-regulate IL-23 as well as IL 12p70 as previous reported. RESULTS: To this end we initially determined that IL 12p70 secretion by control and CD antigen-presenting cells (macrophages) in lamina propria mononuclear populations is optimized by stimulation with CD40L and interferon-gamma. In subsequent studies using these stimulation conditions we found that patients with CD manifested both increased IL-12p70 and IL-23 secretion before anti-IL-12p40 mAb treatment and normal levels of secretion of these cytokines following cessation of treatment. Antigen-presenting cells in lamina propria mononuclear cells from ulcerative colitis patients, in contrast, produced only baseline levels of IL-23. Finally, we found that IL-23-induced T cell production of IL-17 and IL-6 are also greatly reduced after antibody treatment. The latter data are parallel to those from previous studies showing that anti-IL-12p40 down-regulates IFN-gamma and tumor necrosis factor-alpha secretion. CONCLUSIONS: We conclude that CD but not ulcerative colitis is associated with high levels of both IL-12p70 and IL-23 secretion as well as the secretion of downstream effector cytokines, and that this cytokine production is down-regulated following administration of IL-12p40 mAb. PMID- 16374254 TI - Caspase activation and apoptosis induction by adalimumab: demonstration in vitro and in vivo in a chimeric mouse model. AB - BACKGROUND: Adalimumab is a fully human monoclonal antibody to tumor necrosis factor (TNF), which was recently introduced as a therapy for Crohn's disease and rheumatoid arthritis. Besides neutralization, induction of apoptosis of monocytes/macrophages and T cells is thought to be an important mechanism of action of the anti-tumor necrosis factor monoclonal antibody infliximab, at least in Crohn's disease therapy. AIM: To study caspase activation and the induction of apoptosis by adalimumab and the effect of a caspase inhibitor in vivo. METHODS: For in vitro studies, THP-1 cells (human monocytic cell line) were incubated with adalimumab, infliximab, or human immunoglobulin G, and annexin V + propidium iodide, Apo2.7, and 7-amino actinomycin-D were used to study apoptosis on the cell membrane, mitochodrial, and DNA level, respectively. Active caspase-3 was detected by intracellular staining. For in vivo studies, a chimeric human-mouse model was used, in which THP-1 cells were injected intraperitoneally in SCID Beige mice followed by treatment with adalimumab, infliximab, or human immunoglobulin G. Effects of a pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala Asp-fluoromethyketone on apoptosis induction were evaluated. RESULTS: In vitro analysis revealed that apoptosis could be induced in THP-1 cells by both adalimumab and infliximab. Activation of caspase-3 after incubation with adalimumab was demonstrated by intracellular staining. In addition, in the chimeric mouse model, a higher percentage of residual THP-1 cells were apoptotic, and lower cell numbers were recovered in the adalimumab- or infliximab-treated mouse. Apoptosis induction by adalimumab could be abrogated through in vivo pretreatment of mice with the pan-caspase inhibitor. CONCLUSIONS: Adalimumab, besides neutralizing tumor necrosis factor, also induces apoptosis of transmembrane tumor necrosis factor-positive THP-1 cells by activating intracellular caspases. This activity is likely to be important for the clinical effect of this biodrug. PMID- 16374255 TI - Budesonide in the treatment of inflammatory bowel disease: the first year of experience in clinical practice. AB - BACKGROUND: Delayed release budesonide was approved by the FDA for the treatment of mildly to moderately active Crohn's disease involving the ileum and ascending colon. Controlled trials have demonstrated that budesonide is effective in inducing remission and for maintenance of remission, with less frequent steroid side effects than conventional steroids. We sought to determine the benefit of this medication in clinical practice and to identify any non-FDA-approved uses that may warrant further study. METHODS: Patients in whom oral budesonide was prescribed between November 1, 2001 and October 31, 2002, were identified and medical records were reviewed. Patients were categorized by indication for therapy: ileocolonic Crohn's disease (group 1), Crohn's disease elsewhere (group 2), and other conditions (group 3). RESULTS: A total of 225 patients were identified (108 in group 1, 62 in group 2, and 55 in group 3). Group 3 included patients with microscopic colitis (n = 28), pouchitis (n = 13), ulcerative colitis (n = 12), and celiac disease (n = 2). A favorable outcome occurred in 61% of group 1 patients but only 24% of patients in group 2. In group 3, only microscopic colitis patients and pouchitis patients experienced response rates >50% (77% and 60%, respectively). CONCLUSION: Budesonide is effective in a majority of patients with ileocolonic Crohn's disease and microscopic colitis, which is consistent with results reported from clinical trials. A majority of patients with pouchitis also benefit from budesonide therapy, but prospective controlled trials are necessary to clarify the benefit in this group. PMID- 16374256 TI - MDR1 gene: susceptibility in Spanish Crohn's disease and ulcerative colitis patients. AB - BACKGROUND: The multidrug resistance MDR1 gene codes for a membrane transporter associated with inflammatory bowel disease. The polymorphism Ala893Ser/Thr (G2677T/A) previously showed significant association with Crohn's disease (CD) and the Ile1145Ile (C3435T) with ulcerative colitis (UC). We studied the association of both polymorphisms in an independent population to reveal the impact of the MDR1 gene on predisposition to inflammatory bowel disease. METHODS: Case-control study with 321 CD and 330 UC white Spanish patients recruited from the same center, and 352 healthy ethnically matched controls. RESULTS: A significant association of MDR1 C3435T with CD was observed (CC vs (CT + TT): P = 0.007; OR [95% CI] = 1.58 [1.12-2.23]). A CD susceptibility haplotype 2677T/C3435 was identified. No difference between UC patients as a whole and controls could be detected. CONCLUSIONS: New evidence supports the role of the MDR1 gene on CD susceptibility. Therefore, considering our results and those from others, the MDR1 gene behaves as a common risk factor for both CD and UC. We discovered that the C3435 allele conferring susceptibility to CD is different from the described 3435T UC risk allele. PMID- 16374257 TI - Functional gastrointestinal disorders and mood disorders in patients with inactive inflammatory bowel disease: prevalence and impact on health. AB - BACKGROUND: Symptoms of functional gastrointestinal disorders (FGID) associated with mood disorders (MD), such as depression and anxiety, occur in some patients with quiescent inflammatory bowel disease (IBD) and could be caused by changes in gut motility, visceral hypersensitivity, or psychological dysfunction. We assessed the prevalence of FGID symptoms and mood disorders in ambulatory patients with quiescent IBD and examined their impact on health-related quality of life (HRQOL) and use of health resources. METHODS: Consecutive ambulatory patients with IBD completed a survey of Rome II criteria for FGID, the Hospital Anxiety and Depression Survey, HRQOL indices, and health resource utilization. Logistic and linear regression analyses tested for predictors of FGID and HRQOL. RESULTS: Of 361 patients surveyed, 149 (44 ulcerative colitis [UC] and 105 Crohn's disease [CD]) had inactive IBD during the previous 12 months. Symptoms of at least 1 FGID occurred in 81.9%. Functional anorectal disorders were the most prevalent (53.7%) followed by functional bowel disorders (51.7%), and both were of greater prevalence than in the Canadian population (41.6% and 22.6%, respectively). Irritable bowel syndrome symptoms were more common in inactive CD than in UC (26% versus 9.1%, P = .01) and functional constipation was more common in inactive UC than in CD (26.2% versus 5.8%, P < .01). MD occurred in 27.3% of UC and 31.3% of CD patients. Age > or =40 years and anxiety independently predicted an FGID. Both FGID symptoms and MD were associated with impaired HRQOL and increased use of health services. CONCLUSIONS: Many patients with inactive IBD have symptoms compatible with FGID. Both FGID-like symptoms and MD are associated with impaired HRQOL and increased health resource utilization. Recognition and treatment of FGID and MD could potentially improve daily functioning of IBD patients. PMID- 16374258 TI - Impact of chronic conditions on quality of life in patients with inflammatory bowel disease. AB - BACKGROUND: Although studies suggest that inflammatory bowel disease (IBD) has a significant impact on an individual's health-related quality of life, the added weight of other health conditions and comorbidities has not been investigated. The purpose of this study was to expand on prior research by taking into account the impact of other chronic health conditions on the health-related quality of life of individuals with IBD, and to develop a model to help clinicians understand the relative impact of various predictors of their patients' physical and mental health-related quality of life. METHODS: 615 patients from the gastroenterology outpatient practice of a large, urban university hospital received a self-administered survey including questions about their health conditions, the severity of their bowel symptoms, and their health-related quality of life (measured using the SF-36 instrument). RESULTS: 314 completed surveys were returned, resulting in a response rate of 51.1%. Two regression analyses were conducted to identify the role of patient demographic variables and other chronic conditions on the 2 primary outcomes of interest: the SF-36 Physical Component and Mental Component Summary scores. Statistically significant predictors of physical quality of life included IBD disease severity, arthritis, heart disease, age, anemia, back/shoulder pain, and hypertension; statistically significant predictors of mental health-related quality of life were IBD disease severity, depression/anxiety, age, and headaches. CONCLUSIONS: IBD disease severity is the most important predictor of both physical and mental health related quality of life in patients with this condition despite the presence of other chronic conditions. PMID- 16374259 TI - Increased insulin resistance and beta cell activity in patients with Crohn's disease. AB - BACKGROUND: Pancreatitis and exocrine pancreatitic insufficiency have been described as extraintestinal manifestations of inflammatory bowel disease. In this study, we investigated whether the endocrine pancreatic function is also disturbed in patients with inflammatory bowel disease. METHODS: Seventeen patients with Crohn's disease and 13 healthy volunteers participated in the study. We analyzed the plasma insulin response in a 75-g oral glucose tolerance test. Glucose and insulin levels were determined at time 0 (fasting levels) and 30, 60, 90, 120, 180, 240, and 300 min after glucose uptake. Insulin resistance and beta cell function (BCF) were analyzed by calculating respective indices. RESULTS: Fasting and oral glucose-tolerance test glucose levels appeared to be similar in patients with Crohn's disease and in the controls. Impaired fasting glucose, impaired glucose tolerance, and/or overt diabetes mellitus were not observed in the volunteers. Insulin as well as the index for BCF were significantly increased in patients with Crohn's disease. In addition, insulin resistance was shown to be significantly elevated in Crohn's disease. CONCLUSIONS: Patients with Crohn's disease reveal an increased insulin secretion caused by an enhanced BCF, which may be induced by an up-regulated enteropancreatic axis. This hypersecretion may override the insulin resistance given by the chronic inflammatory state. PMID- 16374260 TI - Crohn's disease runs a more aggressive course in young Asian patients. AB - BACKGROUND: Crohn's disease is a heterogeneous inflammatory bowel disease. The impact of age at diagnosis on the clinical course of patients varies widely as reported in the Western literature. Using the Vienna Classification, we seek to determine whether young Crohn's disease patients in an Asian population followed a different clinical course than old patients. METHODS: The case records of 100 Crohn's disease patients who were treated at the Inflammatory Bowel Disease Center, Singapore General Hospital, were studied retrospectively. The age group and location of disease and behavior according to the Vienna classification were determined at diagnosis. RESULTS: A1 group (age <40 years) defined as "young" and A2 group (age > or =40) defined as "old" contained 65 and 35 patients, respectively. Median age for the young group was 27.4 years and that for the old group was 52.6 years. Of the young patients, 66.7% flared at least once compared with 28.6% of the old patients, odds ratio of 5.0 (P < 0.001). Young patients were more likely to be steroid dependent (20.0% of A1 versus 8.6% of A2, P = 0.14), received azathioprine (38.5% of A1 versus 5.7% of A2, P < 0.001) and experienced complications (31% of A1 versus 20% of A2, P = 0.25)-numerically higher rates that did not reach statistical significance. There was no significant difference between the age groups for the location and behavior of disease as well as requirement for surgery. CONCLUSION: In this first Asian study looking specifically at the impact of age at diagnosis of Crohn's disease, we found that young patients underwent a more aggressive clinical course. PMID- 16374261 TI - Specific antibodies against recombinant protein of insertion element 900 of Mycobacterium avium subspecies paratuberculosis in Japanese patients with Crohn's disease. AB - BACKGROUND: Mycobacterial avium subspecies paratuberculosis (MAP) infection has been hypothesized as an etiological factor of Crohn's disease (CD). However, the involvement of MAP in the pathophysiology of CD is controversial. The aim of this study is to investigate whether MAP is involved in the pathogenesis of CD with the glutathione S-transferase fusion recombinant protein encoding a portion of insertion element (IS) 900 (IS900-GST), which is specific for MAP. METHODS: Serum samples from the patients with CD (n = 50), ulcerative colitis (n = 40), colonic tuberculosis (n = 20), and non-IBD controls (n = 44), were applied for solid phase enzyme-linked immunosorbent assay (ELISA) to detect antibodies against MAP and Saccharomyces cerevisiae. IS900-GST, which was made by the pGST-4T-2 vector inserted with polymerase chain reaction-amplified IS900DNA, was used as an antigen of MAP. Moreover, we studied the relationship between antibodies against IS900-GST and clinical characteristics. RESULTS: ELISA showed that the serum level of immunoglobulin G and immunoglobulin A antibodies against IS900-GST (anti IS900) in patients with CD were significantly higher than those with ulcerative colitis, colonic tuberculosis, and control subjects. The levels of anti-IS900 tended to be higher in CD patients with small intestinal involvement than with colonic involvement alone. Anti-IS900 in patients with penetrating- and stricture type CD was significantly higher than with inflammatory-type CD. Furthermore, a negative correlation was found between the titer of anti-IS900 and disease duration. Anti-IS900 was not associated with surgical treatment nor was it associated with the use of immunosuppressants. No significant correlation was observed between the serum levels of anti-IS900 and anti-S cerevisiae antibody. CONCLUSIONS: This is the first demonstration of the ELISA system of detecting antibodies against IS900 in IBD patients. MAP could be involved in the pathophysiology of Japanese patients with CD. PMID- 16374262 TI - Educate, communicate, anticipate-practical recommendations for transitioning adolescents with IBD to adult health care. AB - The cyclical nature of chronic illness requires that children with inflammatory bowel disease be transitioned to the adult medical system with as much continuity of care as possible. Transition from pediatric to adult medical care continues to present significant barriers. The philosophy of transition centers on a process, with the actual transfer to adult care as a point along this clinical pathway. Concrete steps can be taken to help patients prepare for new responsibilities. The aim of this review is to propose a clinical and developmental timeline for both patients and their medical team, including specific responsibilities for both, so that the partnership can best promote a successful transition. PMID- 16374264 TI - To be or not to be: infliximab during pregnancy? PMID- 16374263 TI - From SNPs to haplotypes: unraveling the role of MDR1 in IBD. PMID- 16374265 TI - Thyroid cancer and Crohn's disease: association or coincidence? PMID- 16374266 TI - Leading and managing in unmanageable times. PMID- 16374267 TI - Effect of patient load on trauma outcomes in a Level I trauma center. AB - OBJECTIVE: Increased medical staff workload has been associated with worse outcomes in several studies. Inappropriate staffing has also been implicated in the increased risk of mortality for medical patients admitted on weekends. A theoretical threshold patient load may exist, beyond which the resources are strained and patient outcomes suffer. The goal of the study was to see whether trauma patients admitted during 'high' patient-load periods, at night, or on weekends had worse outcomes. METHODS: Trauma patients admitted to a high-volume Level I trauma center from 1994 to 2002 were analyzed. Patient load was defined as a combination of the number of patients admitted and the severity of their illness. On the basis of a multivariate regression model, a probability of fatal outcome was calculated for each patient as a marker for the severity of illness. For each patient, two new variables were calculated, the number of admissions (#ad) and the average probability of fatal outcome (PFO) for the 24-hour period in which the patient was admitted (excluding the patient him- or herself). The above variables, night/d, and weekend/d were placed in a multivariate regression model. RESULTS: There were 30,686 patients. Age, mechanism of injury, Injury Severity Score, maximum head Abbreviated Injury Scale score, admission Glasgow Coma Scale score, systolic blood pressure, and intubation status were the independent predictors of mortality. This model had an outstanding predictive power, with an area under the receiver operating characteristic curve of 0.96. The mean #ad was 11 +/- 4 and PFO was 0.08 +/- 0.07. Values above the 90th percentile were considered 'high' for #ad > 17 or PFO > 0.18. There was no difference in mortality for patients admitted during high #ad (odds ratio [OR], 0.95; p = 0.7) or high PFO (OR, 0.99; p = 0.9) versus low. There was no difference in mortality if a patient was admitted on weekends versus weekdays (OR, 0.9; p = 0.2) or at night versus day (OR, 0.9; p = 0.2). There was no difference in hospital length of stay for high #ad, high PFO, nights, or weekends. CONCLUSION: At this Level I trauma center that is part of an established statewide trauma system, patient outcomes were not compromised during high-patient-load periods, at night, or on weekends. PMID- 16374268 TI - Manual vital signs reliably predict need for life-saving interventions in trauma patients. AB - OBJECTIVE: Various types of diagnostic and monitoring techniques are available in the prehospital environment. It is unclear how increasing complexity of diagnostic equipment improves the ability to predict the need for a life-saving intervention (LSI). In this study, we determined whether the addition of diagnostic equipment improved the predictive power of vital signs and scores obtained only by physical examination. METHODS: Institutional review board approval was obtained for an analysis of 793 prehospital trauma patient records collected during helicopter transport by Emergency Medical Services personnel. Exclusion of severe head injuries and patients with incomplete data resulted in 381 patients available for analysis. Data sets were classified on the basis of the instrumentation requirements for capturing the given measurements and were defined by three groups: Group 1, vital signs obtained with no equipment (radial, femoral, and carotid pulse character; capillary refill; motor and verbal components of the Glasgow Coma Scale [GCS]); Group 2, Group 1 plus eye component of the GCS and pulse oximetry (Spo(2)); and Group 3, Group 2 plus fully automated noninvasive blood pressure measurements, heart rate, end-tidal carbon dioxide, and respiratory rate. LSIs performed during transport and in the hospital were recorded. Data were analyzed using a multivariate logistic regression model to determine which vital signs were the best predictors of LSI. RESULTS: Radial pulse character and GCS verbal and motor components had the best predictive power for the need of a prehospital LSI in Group 1 (receiver operating characteristic [ROC] curve, 0.97). Radial pulse character together with the eye component of the GCS and the motor component of the GCS provided the best prediction of a need for a prehospital LSI for Group 2 (ROC curve, 0.97). Addition of all supplementary vital signs measured by an automated monitor (Group 3) resulted in an ROC curve of 0.97. Given an abnormal radial pulse character (weak or absent) and abnormal GCS verbal and motor components, the probability of needing an LSI was greater than 88%. CONCLUSION: In this cohort of patients, predicting the need for an LSI could have been achieved from GCS motor and verbal components and radial pulse character without automated monitors. These data show that simple and rapidly acquired manual measurements could be used to effectively triage non-head-injured trauma casualties. Similar results were obtained from manual measurements compared with those recorded from automated medical instrumentation that may be unavailable or difficult to use in the field. PMID- 16374269 TI - Hypotension does not increase mortality in brain-injured patients more than it does in non-brain-injured Patients. AB - OBJECTIVES: Hypotension increases mortality after all types of injuries. Prior studies comparing mortality of hypotensive traumatic brain injury (TBI) patients to normotensive TBI patients have implied that hypotension is particularly detrimental after TBI. It is unknown whether hypotension affects TBI patients more severely than it affects other types of patients. We hypothesized that hypotension does not increase mortality in TBI patients more than it does in non TBI patients. METHODS: National Trauma Data Bank (1994-2002) patients aged 18 to 45 years with blunt mechanisms of injury treated at Level I and Level II centers were included. Deaths occurring before 24 hours were excluded. Logistic regression was used to measure the association between hypotension (< or =90 mm Hg) and death after adjusting for confounding variables of age, gender, comorbidities, complications, Glasgow Coma Scale score, and severity of associated injuries. Odds ratios (95% confidence interval) indicate the risk of death in hypotensive patients in each group compared with normotensive patients in the same group. RESULTS: The study population consisted of 79,478 patients (TBI, 30,742; no TBI, 48,736). Hypotension independently quadrupled the risk of death after adjusting for confounding variables (odds ratio [OR], 4.8; 95% confidence interval [CI], 4.1-5.6). However, increase in this risk associated with hypotension was the same in TBI (OR, 4.1; 95% CI, 3.5-4.9) and non-TBI patients (OR, 4.6; 95% CI, 3.4-6.0). Furthermore, the relationship between hypotension and TBI did not change with increasing head Abbreviated Injury Scale score severity. CONCLUSION: Hypotension is an independent risk factor for mortality. However, it does not increase mortality in TBI patients more than it does for non-TBI patients. PMID- 16374270 TI - A survey of EAST member practices in blunt splenic injury: a description of current trends and opportunities for improvement. AB - BACKGROUND: The literature on blunt splenic trauma provides little evidence-based direction for nonsurgical management. The appropriate role of computed tomography (CT) after initial diagnosis, activity restriction and follow-up are continuing controversies. METHODS: Active EAST members were surveyed regarding in-hospital management and follow-up of patients with isolated and near-isolated blunt splenic injury. Analyses were performed using descriptive and correlational statistics. RESULTS: A 38.4% response rate was obtained. 82.6% of respondents practiced at a Level I trauma centers. 97% of respondents considered hemodynamic instability as the primary indication for immediate splenectomy. 71.6% of respondents preferred ultrasound for initial imaging in hemodynamically stable patients. One-third of respondents admitted stable Grade I patients to monitored settings. 85.5% would not routinely perform predischarge abdominal CT scan in the absence of clinical deterioration, extravasation on initial CT or high-grade injury. Activity restriction varied by grade (table). The majority of respondents (78.1%) relied on clinical judgment alone for activity recommendations in lower grades of injury while a higher reliance on CT was used for Grades IV and V (49.8%). CONCLUSIONS: Despite the lack of evidence-based guidelines, many in hospital and follow-up practices were reasonably consistent. However, some important contradictions were noted (such as monitoring very low risk patients and not intervening in patients with contrast blush). Activity restrictions were usually based on clinical judgment supplemented by CT at the highest grades of injury. Lack of evidence-based guidelines and high reliance on clinical judgment underscore the need for a well-designed prospective study to define optimal management and follow-up. PMID- 16374271 TI - Fascial ultrasound for evaluation of anterior abdominal stab wound injury. AB - BACKGROUND: Local stab wound (SW) exploration to assess abdominal fascial integrity is a highly invasive procedure frequently performed under demanding circumstances in the Emergency Department (ED). We hypothesized ultrasound (U/S) may be useful in the detection of fascial defects resulting from anterior abdominal stab injury, eliminating the need for local wound exploration METHODS: Thirty-five hemodynamically normal patients evaluated at a Level I trauma center for anterior abdominal stab wounds were examined by U/S (8 mHz probe) for evidence of fascial violation. All patients were subsequently evaluated by local wound exploration RESULTS: Fascial U/S had an overall sensitivity of 59% and specificity of 100%, (PPV 100%, NPV 59%) for detection of fascial SW defects compared with local wound exploration. The sensitivity of fascial U/S for stab wound evaluation varied directly with experience of the sonographer CONCLUSIONS: A positive fascial U/S obviates the need for invasive SW exploration; however, a negative fascial U/S does not preclude the need for local wound exploration. Resident U/S training for specific penetrating injuries may reduce the need for abdominal SW fascial exploration in the ED. PMID- 16374272 TI - Removal of the OptEase retrievable vena cava filter is not feasible after extended time periods because of filter protrusion through the vena cava. AB - BACKGROUND: Therapeutic and prophylactic vena cava filters (VCFs) are used to prevent pulmonary embolism. Concerns exist over placing a permanent filter in a young trauma patient. Recently, retrievable VCFs have become available. One such filter is the OptEase, which has a recommended time of removal of up to 23 days after insertion. Data supporting this recommendation are sparse. Many trauma patients will need filters for more than 2 weeks, and there are no data evaluating the safety of removal after extended time periods. The purpose of this study was to determine the safety, feasibility, and reaction of the vena cava when removing the OptEase retrievable VCF at different time intervals. METHODS: Twenty Yorkshire cross pigs (80-113 kg) underwent general anesthesia with tiletamine and zolazepam. Filters were placed in the infrarenal vena cava (VC) through the femoral vein under fluoroscopic guidance. Animals were then divided into four groups. In group 1, filters were removed at 14 days; in group 2, at 30 days; in group 3, at 60 days; and in group 4, at 90 days. Removal was attempted using a snare-and-sheath technique through the femoral vein. Animals with successful filter removal were allowed to recover; then, the animals underwent autopsy (gross and microscopic VC examination) 2 months later. Animals with unsuccessful filter removal underwent autopsy immediately after attempted removal. Venacavograms were taken at filter insertion, at removal, and before autopsy to evaluate any VC abnormalities. RESULTS: Successful removal of the filter in all five pigs (100%) was reliably performed only in the 14-day group. In this group, the initial VC transverse diameter was 19.4 +/- 0.8 mm and was significantly reduced to 9.8 +/- 1.1 mm (p < 0.05) immediately after removal. Sixty days later, before autopsy, VC diameter had increased to 15.3 +/- 1.9 mm, which was significantly larger than at removal (p < 0.05) but not different from the initial value. In the 30-day group, removal was successful in only one of five animals. Although removal was successful in the one pig, autopsy at 2 months postremoval revealed total occlusion of the VC. Filters could not be removed from 60- and 90-day groups. At autopsy, the VCF struts were embedded or protruded through the VC wall. Microscopic examination of the VC revealed significant scarring underneath and between the struts. CONCLUSION: Removal of the retrievable OptEase VCF may be successfully performed up to 14 days after insertion. Strut protrusion through the VC wall prohibited successful and safe removal at extended time intervals. PMID- 16374273 TI - Systemic coagulation changes caused by pulmonary artery catheters: laboratory findings and clinical correlation. AB - BACKGROUND: A higher rate of pulmonary embolism has been associated with pulmonary artery (PA) catheters; however, no mechanism has been described. Conventional tests of coagulation reveal no changes related to PA catheterization. The purpose of this study was to determine whether PA catheterization resulted in a hypercoagulable state detectable by thrombelastography (TEG). METHODS: ANIMAL: Healthy, anesthetized, swine (n = 19) underwent PA catheterization. Samples were drawn from 7F femoral arterial catheters before and two hours after PA catheterization, at 5 mL/min, and analyzed (native whole blood, n = 15, kaolin activated blood, n = 4) by TEG (Hemoscope, Niles, IL) at precisely two minutes. Human: An IRB-approved prospective, observational trial was conducted in critically ill patients (n = 19). Samples were drawn from 22-gauge radial artery catheters, before and three hours after PA catheterization. Kaolin-activated TEG samples were analyzed at precisely five minutes. Data are mean +/- SE; Groups were compared with analysis of variance and significance was assessed at the 95% confidence interval. RESULTS: In both animals and patients, PA catheterization truncated R times (time to initial fibrin formation). In swine, the R times were 17.6 +/- 1.3 minutes (native) and 3.8 +/- 0.4 (kaolin) before PA catheterization, and decreased to 6.3 +/- 1.0 minutes (p = 0.002) and 1.9 +/- 0.5 minutes (p = 0.010) afterward. There were no changes in pH or temperature during the experiment. In patients, 4 of 19 were excluded for protocol violations. The R time was 6.3 +/- 1.0 minutes (kaolin) before and 3.0 +/- 0.3 minutes after catheterization (p = 0.003). No changes were observed in conventional coagulation parameters, temperature or pH. CONCLUSION: In healthy swine, and critically ill patients, PA catheters may enhance thrombin formation and fibrin polymerization, indicating a systemic hypercoagulable state. This may explain why PA catheters are associated with an increased risk of pulmonary emboli. PMID- 16374274 TI - Functional outcome after blunt and penetrating carotid artery injuries: analysis of the National Trauma Data Bank. AB - BACKGROUND: We analyzed the functional outcome at discharge among a cohort of patients suffering traumatic carotid arterial injury (CAI) and compared them with matched controls. METHODS: CAI were identified by International Classification of Diseases-9 codes from the National Trauma Data Bank. Control groups were matched by age, mechanism, head Abbreviated Injury Scale score, Injury Severity Score, base deficit, and admission Glasgow Coma Scale score. Outcome was assessed using the functional independence measure (FIM) score for feeding, locomotion, and expression. FIM scores for each category ranged from 1 (full assistance required) to 4 (fully independent), with a maximum total FIM score of 12 representing full independence. RESULTS: There were 967 CAI identified among 474,024 patients for an overall incidence of 0.2%. The mechanism of injury was blunt in 570 (59%) and penetrating in 397 (41%) patients. Seventy-eight percent of penetrating CAI patients were fully independent for locomotion at discharge (FIM = 4) compared with only 37% of blunt patients (p < 0.01). Compared with the control group of similarly injured patients without carotid injury (n = 14,119), the blunt CAI group demonstrated more severe functional disability (FIM < or = 2) at discharge for feeding, expression, and locomotion. Fifty-five percent of blunt control patients were discharged fully independent (FIM total = 12) compared with only 33% of blunt CAI (odds ratio= 2.5; p < 0.01). The penetrating CAI group showed no significant difference in severe disability for feeding, expression, or locomotion compared with controls. However, 80% of penetrating control patients were discharged fully independent (FIM total = 12) compared with only 67% of patients with penetrating CAI (odds ratio = 2.0; p = 0.01). CONCLUSION: Blunt CAI is associated with more severe functional disability at discharge than penetrating CAI. Even when compared with a similarly injured control group, blunt CAI results in significant additional severe functional disabilities in feeding, expression, and locomotion. PMID- 16374275 TI - Comparison of hemorrhage control agents applied to lethal extremity arterial hemorrhages in swine. AB - BACKGROUND: QuikClot powder (QC), chitosan dressing (CD), and fibrin sealant dressing (FSD) are new hemostatic products touted to be more effective in controlling severe extremity bleeding than the current standard gauze dressing. All have been utilized in the global war on terrorism. Our objective was to evaluate the hemostatic efficacy of these three products in a model of severe extremity arterial hemorrhage that could not be stopped by standard gauze treatment. METHODS: A model of severe extremity arterial hemorrhage was developed in swine that was 100% fatal with standard gauze application and manual compression. The Army Field Bandage (AFB) was the standard gauze control. Anesthetized animals (n = 60, 15/group, 37.7 +/- 2.5 kg) were splenectomized and instrumented. A reproducible femoral artery injury was created using a 6 mm aortic punch, and free bleeding was allowed for 45 seconds. Each hemostatic agent was applied twice with three-minute compressions. All products were applied on actively bleeding wounds through a pool of blood. Fluid resuscitation was started with the first compression and titrated to a mean pressure of 65 mm Hg. Animals were observed for 180 minutes or until death. Endpoints were percent survival, survival time, blood loss, resuscitation volume, wound temperatures and tissue histology. Data are expressed as mean +/- SD and analyzed by Fisher's exact, logrank, and nonparametric ANOVA tests. RESULTS: Baseline physiologic parameters were similar among groups. AFB did not produce hemostasis. QC also showed no hemostatic benefit, and QC treatment markedly increased maximum wound temperatures to an average of 70.8 +/- 4.2 degrees C (p < 0.001). CD stopped bleeding temporarily in only one animal. There were no survivors in the AFB, QC, or CD groups. CD numerically prolonged survival time (58.9 +/- 21.1 minute) compared with the control (38.4 +/- 24.7 minutes, p = 0.045) but the difference was not significant. FSD reduced bleeding (p < 0.05) and prevented exsanguination in 10/15 (2/3) animals, and resulted in a significantly longer average survival time (p < 0.0001). CONCLUSION: FSD was superior to other currently utilized hemostatic products in controlling lethal arterial hemorrhage in this model of a fatal extremity wound. CD showed some hemostatic benefit. The exothermic reaction of QC was significant and resulted in gross and histologic tissue changes of unknown clinical significance. Controlled human studies with the promising products are required. PMID- 16374276 TI - Early vasopressin improves short-term survival after pulmonary contusion. AB - Arginine vasopressin (AVP) is a promising treatment for several types of irreversible shock, but its therapeutic potential has not been examined after severe chest trauma. Two series of experiments were performed to fill this gap. METHODS: Series 1: anesthetized, mechanically-ventilated pigs (n = 20, 29 +/- 1 kg) received a blast to the chest, followed by a "controlled" arterial hemorrhage to a mean arterial pressure (MAP) <30 mm Hg. At 20 minutes, a 10 mL/kg normal saline (NS) bolus was followed by either 0.1 U/kg AVP bolus or NS, in randomized, blinded fashion. From 30-300 minutes, either AVP (0.4 U/kg/hr plus NS) or NS alone was infused as needed to MAP>70 mm Hg. Series 2: Swine (n = 15) received the chest injury followed by partial left hepatectomy to produce "uncontrolled" hemorrhage. Resuscitation was the same as in series 1. RESULTS: The blast created bilateral parenchymal contusions (R > L), hemo/pneumothorax and progressive cardiopulmonary distress. In Series 1, there were 3/20 deaths before randomization, 0/8 deaths after resuscitation with AVP versus 4/9 deaths with NS (p = 0.029). In surviving animals, with AVP versus NS, fluid requirements and peak airway pressures were lower while P/F was higher (all p < 0.05). In Series 2, with uncontrolled hemorrhage, there were 5/15 deaths before randomization. Upon resuscitation with AVP versus NS, survival time and blood loss were both improved, but the differences did not reach statistical significance. CONCLUSIONS: After severe chest trauma with controlled hemorrhage, early AVP decreased mortality, reduced fluid requirements and improved pulmonary function. With uncontrolled hemorrhage, early AVP did not increase the risk for bleeding. PMID- 16374277 TI - The impact of a hypercatecholamine state on erythropoiesis following severe injury and the role of IL-6. AB - BACKGROUND: Severe traumatic injury can lead to hemorrhagic shock-induced bone marrow (BM) dysfunction resulting in persistent anemia. The hypercatacholamine state that accompanies severe injury has been shown to impact the growth of erythroid progenitors. IL-6 has a role both in the acute phase response of trauma and has been implicated in the development of anemia. The aim of this study was to investigate the severity of a hyper-adrenergic stimulus on pluripotent progenitors (GEMM-CFU) as well as erythroid progenitors (BFU-E and CFU-E) and the potential regulatory role of IL-6. METHODS: Normal human BM mononuclear cells were isolated and erythropoiesis was assessed by the growth of GEMM-CFU, BFU-E and CFU-E in the presence of adrenergic agonists, norepinephrine (NE) and epinephrine (EPI), at increasing concentrations. Similarly, normal BM stroma cells were grown to confluence then incubated with NE and EPI. Supernatant was harvested and IL-6 levels were determined using ELISA. RESULTS: Under physiologic conditions (10(-7) M), NE and EPI increase BFU-E and CFU-E growth (374% and 177% versus 100% control). At severe stress levels (10(-3) M), NE and EPI completely inhibited BFU-E and CFU-E growth (5% and 4% versus 100% control). GEMM-CFU growth was increased by NE and not EPI at 10(-7) M. The presence of NE and EPI increased IL-6 levels in a dose-dependent fashion. CONCLUSIONS: The proliferative effect of adrenergic agonists at physiologic levels on normal erythropoiesis begins early during erythroid differentiation. At severe stress levels, BFU-E and CFU-E growth is inhibited. The erythropoietic dysfunction and resultant anemia seen following severe injury may be due to the presence of a severe hypercatecholamine state and may be mediated by IL-6. PMID- 16374278 TI - Prospective comparison of bronchoalveolar lavage and quantitative deep tracheal aspirate in the diagnosis of ventilator associated pneumonia. AB - BACKGROUND: Ventilator associated pneumonia (VAP) is common in trauma patients, and accurate diagnosis of VAP may improve survival. With the risk of development of bacterial resistance, we also strive to minimize the use of unnecessary antibiotics. Recent studies suggest that quantitative deep endotracheal aspirate (QDEA) is adequate in VAP diagnosis. We currently use bronchoalveolar lavage (BAL) diagnosis. The purpose of this study was to examine the accuracy of QDEA as compared with BAL in diagnosing VAP in trauma patients. METHODS: We prospectively compared the results of BAL and QDEA in intubated patients suspected of having VAP during an 8-month period. Indication for BAL was pulmonary infiltrate, systemic inflammatory response syndrome, and C-reactive protein >17 mg/dL at > or =48 hours after admission. Study patients underwent QDEA immediately before BAL, and quantitative cultures were compared for both specimens. The techniques differ in that QDEA involves the direct culture of sputum suctioned from the distal trachea, whereas BAL involves lavage of the bronchoalveolar tree with sterile saline, which is then cultured. VAP was diagnosed on BAL if > or =10(5) cfu/mL was present on culture. The ability of QDEA to diagnose pneumonia was examined at cutoffs of > or =10(5) cfu/mL and > or =10(4) cfu/mL, as compared with BAL at > or =10(5). RESULTS: Sixty-one patients underwent BAL during this period, and 39 of these underwent both BAL and QDEA for the study. Of the 39 studied patients between March 16, 2002, and November 4, 2002, 20 (51%) were found to have VAP by BAL (> or =10(5) cfu/mL). Using this cutoff for QDEA, 18 of 20 (90%) would have been correctly diagnosed. Using > or =10 cfu/mL for QDEA, the rate of correct diagnosis would increase to 19 of 20 (95%). However, of the 19 who did not have pneumonia according to BAL, 6 (31%) would have been incorrectly diagnosed with VAP using the QDEA cutoff of > or =10(5) cfu/mL. A QDEA cutoff of > or =10 (4) cfu/mL would result in the even higher false-positive rate of 8 of 19 patients (42%). CONCLUSION: Whereas most patients with pneumonia by BAL would have been diagnosed by QDEA, use of QDEA in treatment decisions would have led to needless antibiotic administration in 31% of VAP-negative patients at a cutoff of > or =10(5) cfu/mL and 42% at > or =10(4) cfu/mL. The use of QDEA in VAP diagnosis is limited because of the rate of over-diagnosis. With the increasing problems associated with excess antibiotic use, we believe these results support the use of BAL over QDEA in the diagnosis of VAP in the ventilated trauma patient. PMID- 16374279 TI - The early work-up for isolated ligamentous injury of the cervical spine: does computed tomography scan have a role? AB - BACKGROUND: Helical computed tomography (HCT) scan is the preferred modality for diagnosing fractures of the cervical spine in blunt trauma. We hypothesize that HCT can be used as a screening tool for isolated ligamentous injury (LI) in blunt trauma. METHODS: A prospective, consecutive series study design was used to include patients that could not have their cervical spine cleared clinically. All patients underwent HCT (occiput-T1) and plain radiographs (PR) with five views of the cervical spine. Patients with clinical or radiographic abnormalities without fracture underwent cervical magnetic resonance imaging (MRI). Demographic and outcome data were collected. The attending radiologist's interpretation was used for clinical management. Three neuroradiologists in a blinded fashion re-reviewed the studies (HCT, PR, and MRI) of the MRI subgroup. RESULTS: One thousand five hundred seventy-seven patients met the study criteria. Two hundred seventy-eight had 416 cervical spine fractures. PR failed to identify 299 of 416 (72%) cervical spine fractures in 208 of 278 (74.8%) patients. Of the 1,299 (82%) patients who had no fracture, 85 (6.5%) required an MRI. The mean time from admission to MRI was 3 days for the LI subgroup. Of these, 21 of 85 (25%) had LI by MRI. Seven of 21 (33.3%) patients had an abnormal HCT versus 3 of 21 (14.3%) patients who had an abnormal PR. Four of 85 (4.7%) patients had spinal cord injury without radiographic abnormality. One (1.2%) patient required surgical stabilization of LI, as seen on all studies performed (PR, HCT, and MRI). Sensitivities for PR and HCT for LI were 16% and 32%, respectively. Negative predictive values for PR and HCT for LI were 74% and 78%, respectively. Measurements of interrater reliability for MRI, HCT, and PR had kappa values of 0.60, 0.14, and 0.41, respectively. CONCLUSION: HCT is the most sensitive, specific, and cost-effective modality for screening the cervical spine bony injuries, but it is not an effective modality for screening for cervical LI. MRI is clearly superior to HCT for LI. The indications for MRI include abnormalities on HCT, neurologic deficits, cervical pain or tenderness on examination, or the inability to clear the cervical spine in the obtunded patient. With the current state of the art technology, we have redefined the definition of spinal cord injury without radiographic abnormality to include spinal cord injuries without boney injuries or LI. PMID- 16374280 TI - Esophageal dysfunction in cervical spinal cord injury: a potentially important mechanism of aspiration. AB - BACKGROUND: Respiratory complications are a major cause of morbidity and mortality in patients with cervical spinal cord injury (CSCI). We hypothesized that patients with CSCI had esophageal dysfunction, predisposing them to aspiration. The purpose of this study was to characterize esophageal function in these patients. METHODS: CSCI and similarly injured control (spinal cord injury below T1) subjects were prospectively enrolled from two trauma centers. All underwent esophageal manometry to measure lower (LES) and upper esophageal sphincter (UES) pressures. A subset of patients had detailed manometry and 24 hour pH studies performed to evaluate dynamic esophageal function. RESULTS: Eighteen CSCI and five control subjects were enrolled. The groups were similar with regards to age, sex, injury mechanism, Injury Severity Score, and hospital stay. Resting LES and UES pressures were similar in CSCI and control patients and did not differ from established norms. Five CSCI and two control patients underwent detailed manometric assessment. Defective UES relaxation was observed in all CSCI patients but not controls. CSCI patients had increased UES relaxation pressures (18.4 +/- 5.3 versus 3.9 +/- 0.7 mm Hg; p = 0.01) and UES bolus pressures (23.8 +/- 2.2 versus 10.2 +/- 6.9 mm Hg; p = 0.006) compared with controls. Esophageal body and LES function were normal. Two of five CSCI patients had abnormal 24-hour pH studies. CONCLUSION: Patients with CSCI demonstrate significantly disturbed dynamic function of the pharynx and UES while resting parameters remain normal. Because adequate UES relaxation is critical to the clearance of secretions and coordination of swallowing, this is an important potential mechanism of aspiration in patients with CSCI. PMID- 16374281 TI - Characterizing the need for mechanical ventilation following cervical spinal cord injury with neurologic deficit. AB - BACKGROUND: Patients who sustain cervical spinal cord injury (C-SCI) with neurologic deficit may require a definitive airway and/or prolonged mechanical ventilation. The purpose of this study was to characterize factors associated with a high risk for respiratory failure and/or the need for mechanical ventilation in C-SCI patients. METHODS: Patients with C-SCI and neurologic deficit admitted to a Level I Trauma Center between July 1, 2000 and June 30, 2002 were retrospectively reviewed for demographics, level and completeness of neurologic deficit, need for definitive airway, need for tracheostomy, need for mechanical ventilation at hospital discharge (MVDC), and outcomes. The level and completeness of injury were defined by American Spinal Injury Association standards. RESULTS: One hundred nineteen patients with C-SCI and neurologic deficit were identified over this period. Of these, 45 were identified as complete C-SCI: 12 (27%) patients had levels of C1 to C4; 19 (42%) had a level of C5; and 14 (31%) had levels of C6 and below. There were 37 males and 8 females. There were 36 blunt and 9 penetrating injuries. The average age of these patients was 40 +/- 21, and the average ISS was 45+/-22. Eight of the patients with complete C-SCI died, for a mortality of 18%. Of the 37 survivors, 92% received a definitive airway, 81% received tracheostomy, and 51% required MVDC. All patients with complete injuries at the C5 level and above required a definitive airway and tracheostomy, and 71% of survivors required MVDC. Of the patients with complete injuries of C6 and below, 79% received a definitive airway, 50% required tracheostomy, and 15% of survivors required MVDC. Only 35% of incomplete injuries required a definitive airway, and only 7% required tracheostomy. CONCLUSIONS: The need for definitive airway control, tracheostomy, and ventilator dependence is significant, especially for patients with high complete C-SCI. Based on these results we recommend consideration of early intubation and tracheostomy for patients with complete C-SCI, especially for those with levels of C5 and above. PMID- 16374282 TI - Incidence, risk factors, and outcomes for occult pneumothoraces in victims of major trauma. AB - BACKGROUND: The supine anteroposterior chest radiograph (CXR) is an insensitive test for posttraumatic pneumothoraces (PTXs). Computed tomographic (CT) scanning often detects PTXs that were not diagnosed on CXR (occult PTXs [OPTXs]). The purpose of this study was to define the incidence, predictors, and outcomes for OPTXs after trauma. METHODS: Thoracoabdominal CT scans and corresponding CXRs of all trauma patients entered into a regional database were reviewed. Patients with OPTXs were compared with those with overt, residual, and no PTXs regarding incidence, demographics, associated injuries, early resuscitative predictors, treatment, and outcomes. RESULTS: Paired CXRs and CT scans were available for 338 of 761 (44%) patients (98.5% blunt trauma). One hundred three PTXs were present in 89 patients, 57 (55%) of which were occult; 6 (11%) were seen only on thoracic CT scan. Age, sex, length of stay, and survival were similar between all groups. OPTXs and PTXs were similar in comparative size index and number of images. Subcutaneous emphysema, pulmonary contusion, rib fracture(s), and female sex were independent predictors of OPTXs. Seventeen (35%) patients with OPTXs were ventilated, of whom 13 (76%) underwent thoracostomy. No complications resulted from observation, although 23% of patients with thoracostomy had tube-related complications or required repositioning. CONCLUSION: OPTXs are commonly missed both by CXR and even abdominal CT scanning in seriously injured patients. Basic markers available early in resuscitation are highly predictive for OPTXs and may guide management before CT scanning. Further study of OPTX detection and management is required. PMID- 16374283 TI - Limitations of splenic angioembolization in treating blunt splenic injury. AB - BACKGROUND: When angiography is performed in all hemodynamically normal patients with splenic injury, only 30% require embolization. This study examines the use of selective splenic angioembolization (SAE) as part of a management algorithm for adult splenic injury. METHODS: Criteria for selective SAE were added to our adult splenic injury protocol in July 1999. SAE was performed in hemodynamically stable patients if computed tomographic (CT) scan revealed injury to the hilum or vascular blush and when nonoperative patients had a gradual decrease in hematocrit. Patients were grouped by management strategy: nonoperative; operative; or SAE. Demographics, injury severity, and outcomes of the different groups were compared. Medical records, CT scans, and registry data were reviewed for all SAE cases, deaths, and treatment failures. Data are means +/- SE. p < 0.05 versus nonoperative management by analysis of variance. RESULTS: From July 1999 to August 2003, 194 adults were treated for splenic injury. Nine patients underwent SAE, six for CT findings (1 vascular blush) and three for decreasing hematocrit. Three patients failed SAE (33%), one for bleeding and two for delayed splenic infarction. Eleven patients failed nonoperative therapy (8%); splenorrhaphy was performed in three and splenectomy in eight. Operative patients were more seriously injured and had higher Injury Severity Scores and mortality; splenectomy (39 of 48) was more commonly performed than splenorrhaphy (9 of 48) in this group. CONCLUSION: Use of a splenic injury algorithm is associated with a high success rate for nonoperative management of splenic trauma. Using selective criteria, only 5% of patients were treated with SAE. SAE salvaged six patients with high-grade splenic injury or decreasing hematocrit but had a 33% failure rate. Failure of nonoperative management was most commonly caused by errors in judgment, primarily recognition of "high-risk" injury patterns on CT scan or attempting nonoperative management in anticoagulated or coagulopathic patients. PMID- 16374284 TI - Real-time, contrast-enhanced sonography: a new tool for detecting active bleeding. AB - BACKGROUND: Active contrast medium extravasation is a known angiographic and computed tomographic sign of ongoing, potentially life-threatening hemorrhage. Sonography (US) is frequently the first imaging option for screening patients with abdominal emergencies. Because of the current possibilities of low mechanical-index, real-time, contrast-specific systems, it is possible to detect contrast leakage by using US. The purpose of this article is report our pilot experience in the evaluation of active traumatic and nontraumatic bleeding with contrast-enhanced US. METHODS: In a 2-year period, we performed 153 consecutive emergent contrast-enhanced US studies. Traumatic emergencies accounted for 83 examinations and nontraumatic emergencies accounted for 70. We used the contrast specific mode Contrast Tuned Imaging and the contrast medium SonoVue. A 2.4- to 4.8-mL contrast medium bolus was injected with continuous US acquisition, starting immediately after contrast injection and lasting 1 to 6 minutes. RESULTS: Contrast extravasation was found in 20 cases (13%). These included spleen injury (n = 8), liver injury (n = 3), kidney injury (n = 1), abdominal aortic aneurysm rupture (n = 5), splenic angiosarcoma rupture (n = 1), postsurgical bleeding after abdominal aortic aneurysm repair (n = 1), and postsplenectomy bleeding (n = 1). Active extravasation appeared as a round, hyperechoic pool or as a fountain-like, hyperechoic jet. CONCLUSION: Our retrospective clinical study shows for the first time how US can detect contrast medium extravasation, a significant indicator of active hemorrhage and of need for prompt surgical or interventional treatment. PMID- 16374285 TI - Use of a refined operative strategy in combination with the multidisciplinary approach to manage blunt juxtahepatic venous injuries. AB - BACKGROUND: Despite continuous advances in traumatology, juxtahepatic venous injuries are still the most difficult and deadly form of liver trauma. Most deaths result from exsanguination, and reported mortality ranges from 50% to 80%. This is an evaluation on our experience with the management of this high mortality injury following a refined operative strategy. METHODS: This is a retrospective study of consecutive patients sustaining blunt juxtahepatic venous injuries. The management for these patients was mainly a refined operative strategy combined with a multidisciplinary approach. Preoperative conditions and the patient demographics were gathered. In addition, the number and type of interventional procedures, overall complications, and operative procedures were collected and analyzed. RESULTS: From January, 1996 to March, 2004, 19 patients (M:F = 13:6) with juxtahepatic venous injuries were included and all were managed operatively. The operative procedures included hepatectomy by finger fracture technique for direct repair (8), perihepatic packing (1), packing and hepatic artery embolization (1), packing and hepatic artery ligation (1), hepatorrhaphy and packing (5), packing followed by hepatectomy (2) and atriocaval shunt for direct repair (1). The survival rate for the packing group was higher than that of the direct repair group (75% versus 45%), but was not statistically significant (p = 0.352). Injury to the retrohepatic vena cava influenced the patient's survival significantly (p = 0.041). The overall survival was 58% (11/19). CONCLUSION: A well-defined operative strategy helps surgeons deal with the problem of blunt juxtahepatic venous injury, and its combination with multidisciplinary management will improve patient outcomes. PMID- 16374286 TI - Incidence of and risk factors for acute stress disorder in children with injuries. AB - BACKGROUND: To assess the incidence of and risk factors for Acute Stress Disorder (ASD) in children with injuries. Numerous studies have documented the increased incidence of PTSD in those initially diagnosed with ASD. PTSD symptoms cause tremendous morbidity and may persist for many years in some children. METHODS: Children hospitalized with one or more injuries were interviewed and assessed with the following: Child Stress Disorders Checklist (CSDC), Family Strains Scale, Brief Symptom Inventory (BSI) and Facial Pain Scale. RESULTS: Participants included sixty-five children (ages 7-18 years). The mechanisms of injury varied (e.g. MVC, penetrating). The mean injury severity score was 8.9 +/- 7. The mean length of hospital stay was 4.6 +/- 4.6 days. Altogether, 18 (27.7%) of participants met DSM IV criteria for ASD during their acute hospital stay. Risk factors such as level of family stress, caregiver stress, child's experience of pain, and child's age were predictive of acute stress symptoms. CONCLUSION: We have identified four risk factors of ASD that have implications for the treatment, and possibly, preventative intervention for PTSD. Further investigation and greater understanding of risk factors for ASD in children with injuries may facilitate the design of acute interventions to prevent the long term negative outcomes of traumatic events. PMID- 16374287 TI - Developing a decision instrument to guide computed tomographic imaging of blunt head injury patients. AB - BACKGROUND: Computed tomographic (CT) head scanning of blunt trauma patients is expensive, delays care, and necessitates radiation exposure, while detecting intracranial injuries in a minority of patients. Clinical characteristics may be able reliably identify patients who do not have intracranial injuries and consequently, do no require imaging. METHODS: Physicians assessed blunt trauma patients undergoing imaging for the presence or absence of specific criteria. Recursive partitioning was used to identify criteria that predict intracranial injuries with high sensitivity. RESULTS: Intracranial injuries were found in 917 of 13,728 enrolled patients (6.7%). Injuries were rare among patients under age 65 who had no evidence of skull fracture, scalp hematoma, neurologic deficit, abnormal alertness, abnormal behavior, coagulopathy, or persistent vomiting. These characteristics would have identified 901 injury cases (sensitivity 98.3% [CI: 97.2-99.0]), while classifying 1,752 patients (12.8%) as "low risk." CONCLUSIONS: Clinical characteristics can reliably identify patients who are unlikely to have intracranial injuries and who do not require CT imaging. PMID- 16374288 TI - Determination of significant parameters for eye injury risk from projectiles. AB - BACKGROUND: Eye injuries affect a large proportion of the population and are expensive to treat. This article presents a parametric analysis of experimental data to determine the most significant factors for predicting ocular injuries or tissue lesions. METHODS: Using logistic regression, statistical values were generated to determine significant projectile characteristics for predicting ocular injury in published studies. Projectiles included BBs, metal rods, and foam particles with velocities ranging from 2 m/s to 122 m/s. RESULTS: A normalized energy (energy per projected area) value was found as the best predictor for ocular injury. Using this predictor, a 50% injury risk of corneal abrasion, lens dislocation, hyphema, retinal damage, and globe rupture was found to be 1,503 kg/s(2), 19,194 kg/s(2), 20,188 kg/s(2), 30,351 kg/s(2), and 23,771 kg/s(2), respectively. CONCLUSION: Normalized energy was the most significant predictor of injury type and tissue lesion. This finding is of great value for history-taking management triaging and as a design aid to minimize the risk of ocular injury for consumer products. PMID- 16374289 TI - Topical becaplermin improves outcomes in work related fingertip injuries. AB - BACKGROUND: Fingertip injuries are common and bear significant costs associated with treatment, lost work, and functional impairment. This study compared these factors in occupationally related fingertip injuries treated with becaplermin, a recombinant human platelet-derived growth factor, and those treated with surgical reconstruction. METHODS: This was a prospective controlled trial involving occupationally related fingertip injuries. Fifty men (ages 23-51) with full thickness, single fingertip injuries > or =1.5 cm(2) with or without phalangeal exposure and distal to the distal interphalangeal (DIP) joint were evaluated. Group I (n = 25) underwent treatment with daily topical becaplermin. Group II (n = 25) underwent surgical reconstruction with a skin graft or local soft tissue flap. Time to wound healing, time to return to work, associated treatment costs, and calculated functional impairment were recorded. RESULTS: Patients in Group I returned to work in significantly less time than those in Group II-10 days versus 38 days respectively). The average calculated functional impairment in Group I was 10% versus 22% in Group II. Associated treatment costs in group A were 1580 +/- 145 US Dollars compared with 6750 +/- 785 US Dollars in Group II. All differences were statistically significant at p < 0.05 CONCLUSION: In this study, the functional and economic costs were significantly less when fingertip injuries were treated with topical becaplermin than when they were treated with surgical reconstruction. This information should allow emergency and acute care physicians to treat these injuries more efficaciously and conveniently. PMID- 16374290 TI - The value of washers in cannulated screw fixation of femoral neck fractures. AB - BACKGROUND: Given the limited evidence to support the technical aspects of screw placement for treatment of femoral neck fractures, we conducted an observational study to evaluate demographic and radiographic variables associated with fixation failure. METHODS: Eligible patients with femoral neck fractures were treated with multiple cannulated screws across three academic centers during a 6-year period. The following variables were evaluated for their predictive value for fixation failure: age, gender, fracture type, presence of comminution, total number of screws, the absence of a washer, the screw configuration, reduction quality, the distance of the most inferior screw to the inferior neck, and screw alignment. Variables were evaluated separately and in a multivariable regression model. RESULTS: Eighty patients were included in the study. The overall failure rate was 30%. We identified four variables associated with fixation failure. These included the lack of washers (odds ratio [OR], 11.2; p = 0.03), imperfect quality of reduction (OR, 9.7; p < 0.01), age greater than 75 years (OR, 5.1; p = 0.04), and displaced versus undisplaced fracture type (OR, 3.8; p < 0.01). These four variables accounted for 43% of the variability in fixation failure (R(2) = 0.43). All other variables including the distance of the most inferior screw to the inferior/medial neck were found to be not significant. CONCLUSION: This study confirms previous findings in the literature that increased age, a displaced fracture type, and poor reduction increase the risk of fixation failure. Contradictory to current belief, there was no significant association between the distance of the inferior screw to the inferior/medial femoral neck cortex and fixation failure. A novel finding of the present study is that the use of washers significantly decreases the risk of fixation failure. PMID- 16374291 TI - Tranexamic acid does not prevent rebleeding in an uncontrolled hemorrhage porcine model. AB - BACKGROUND: Fluid resuscitation after uncontrolled hemorrhage might promote rebleeding and irreversible shock. Tranexamic acid is a procoagulant drug that limits blood loss after surgery of the hip, knee, and heart. We hypothesized that pretreatment with tranexamic acid reduces the rebleeding in uncontrolled hemorrhage and thereby allows safe administration of crystalloid fluid resuscitation. METHODS: A 120-minute intravenous infusion of 100 mL/kg of Ringer's solution was given to 24 pigs (mean weight, 20 kg) 10 minutes after lacerating the infrarenal aorta. The animals were randomized to receive an intravenous injection of 15 mg/kg of tranexamic acid or placebo just before starting the resuscitation. Rebleeding events were monitored by two ultrasonic probes positioned proximal and distal to the laceration. RESULTS: Tranexamic acid had no effect on the number of rebleeding events, bled volume, or mortality. The initial bleeding stopped within 4 minutes after the injury. The five animals that died suffered from 4.4 rebleeding events on average, which tripled the total blood loss, whereas the survivors had only 1.3 such events during fluid resuscitation (p < 0.02). At autopsy, death was associated with a larger total hemorrhage; the blood recovered from the abdomen weighed 1.4 kg (median) in nonsurvivors and 0.6 kg in survivors (p < 0.001), with the difference being attributable to rebleeding. CONCLUSION: Rebleeding events increased the amount of blood lost and the mortality in uncontrolled aortic hemorrhage. Tranexamic acid offered no benefit. PMID- 16374292 TI - A low blood ethanol level is associated with improved cytokine production in aged mice after traumatic injury. AB - BACKGROUND: Many trauma patients have been consuming alcohol at the time of injury. Although high concentrations of alcohol are correlated with poor outcome, few studies have examined the effects of low levels of alcohol. We examined the effects of low alcohol exposure after burn injury using a murine model. METHODS: Three- and 18-month-old mice were given ethanol or saline 30 minutes before a 15% total body surface area burn injury. Twenty-four hours after injury, cellular immune responses, including delayed-type hypersensitivity response and splenocyte proliferation were examined, along with production of interferon-gamma, interleukin (IL)-2, and IL-4. RESULTS: Alcohol administration resulted in a significant increase in interferon-gamma in the aged, but not young, burn-injured mice. Likewise, slight increases in IL-2, IL-4, and the delayed-type hypersensitivity response were observed. CONCLUSION: Low levels of ethanol at the time of injury are associated with partial restoration of immune responses in aged mice. PMID- 16374294 TI - 'The hemostat wrap' a new technique in splenorraphy. PMID- 16374293 TI - The 2004 Fitts Lecture: current perspective on combat casualty care. PMID- 16374295 TI - Versatility of the abductor hallucis muscle as a conjoined or distally-based flap. AB - Soft tissue coverage of the medial ankle and foot remains a difficult, challenging, and often frustrating problem to patients as well as surgeons. To our knowledge, the abductor hallucis muscle flap is not frequently used and only a few well documented cases were found in literature. The purpose of this paper is to report and to present the long-term results of a series of four patients who underwent reconstruction of foot and ankle defects with the abductor hallucis muscle flap. In two cases, the abductor hallucis muscle flap was transposed in combination with a medialis pedis flap to cover a medial ankle defect, whereas in another case it was combined with a medial plantar flap. In this latter case, the muscle flap served to fill up a calcaneal dead space after osteomyelitis debridement, whereas the cutaneous flap was used to replace debrided skin at the heel. The abductor hallucis flap was used as a distally-based turnover flap to cover a large forefoot defect in a fourth case. Follow-up period ranged between 18 and 64 months (mean 43.3). In the early postoperative period, two flaps healed completely In two patients marginal flap necrosis occurred which was subsequently skin grafted. No donor-site complication occurred in any of the patients. In all cases, protective sensation of the skin was satisfactory as early as 6 months. In two cases mild hyperkeratosis at the skin graft border to the sole skin (non weight bearing area of medial plantar and medialis pedis flap donor site) was present, but probably related to poor foot care. All patients were fully mobile as early as 3 months after treatment. In the long-term follow-up (43.3 months), all flaps provided with durable coverage. Functional gait deficit due to consumtion of the abductor hallucis muscle was not apparent. Our long-term results demonstrated that the abductor hallucis muscle flap is a versatile, and reliable flap suitable for the reconstruction of foot and ankle defects. Utilizing the abductor hallucis muscle as a pedicled flap (distally or proximally based) with or without conjoined regional fasciocutaneous flaps offers a successful and durable alternative to microsurgical tree flaps for small to moderate defects over the calcaneus region, medial ankle, medial foot, and forefoot with exposed bone, tendon, or joint. PMID- 16374296 TI - Gunshot wound to the left ventricle with bullet embolization to the descending aorta: combined endovascular and surgical management. PMID- 16374297 TI - Complete transection of the common bile duct after blunt trauma: a case report. PMID- 16374298 TI - Percutaneous thrombin injection to treat a post-traumatic hepatic pseudoaneurysm. PMID- 16374299 TI - Combined percutaneous and angiographic thrombosis of a traumatic hepatic artery pseudoaneurysm in a child. PMID- 16374300 TI - Misplacement of central vein catheters in patients with hemothorax: a new approach to resolve the problem. AB - BACKGROUND: In emergency and pre-hospital care, the verification of the correct position of a central venous catheter is based on the observation of blood color reflow as well as pressure changes with respiration. However, in trauma patient with hemothorax, these indices may not always be reliable signs as the catheter is in a blood-filled pleural space. METHODS: A review of reports published describing patients presenting hemothorax and equipped with central venous catheter wrongly assumed to be in the correct position was performed. RESULTS: Over 10 years, seven reports have been published and a last study was found in the references list of one of the reviews. CONCLUSION: In patients with hemothorax due to severe thoracic trauma or other causes, a delay in detection of incorrect placement of a central venous catheter may delay fluid resuscitation and decrease the chances of survival. In this situation, the use of portable ultrasound devices may be an useful method to increase success rate in catheter insertion. PMID- 16374301 TI - Intramural hematoma on acute phase: diagnostic pitfall. PMID- 16374302 TI - Spine injuries following isolated truncal, head, or neck gunshots. PMID- 16374303 TI - Shoulder injuries have been noted as a recurring problem in skydiving. PMID- 16374304 TI - Use of the physiochemical acid-base approach for mortality prediction in trauma intensive care unit patients. PMID- 16374305 TI - Acute care surgery. PMID- 16374307 TI - Preface. PMID- 16374308 TI - Challenges associated with clinical trials for inherited and orphan retinal diseases. PMID- 16374309 TI - Strategies for success in drug development. PMID- 16374310 TI - Attracting pharmaceutical companies as partners in drug development. PMID- 16374311 TI - An overview: attracting partners in the pharmaceutical industry. PMID- 16374312 TI - Hurdles and opportunities for a venture capitalist investing in therapies for orphan retinal diseases. PMID- 16374313 TI - Crucial factors in commercial success for ophthalmic drugs. PMID- 16374314 TI - Key considerations for seeking product development funding. PMID- 16374315 TI - Business modeling. PMID- 16374316 TI - Role of venture capitalists in funding clinical trials. PMID- 16374317 TI - Photoreceptor cell rescue in inherited and orphan retinal diseases: disease specific requirements. PMID- 16374318 TI - Beyond basic research for inherited and orphan retinal diseases: successes and challenges. PMID- 16374319 TI - Cellular mechanisms of retinal degenerations: RPE65, ABCA4, RDS, and bicarbonate transporter genes as examples. PMID- 16374320 TI - Effect of gene expression on cone survival in retinitis pigmentosa. PMID- 16374321 TI - Survival factors for treatment of retinal degenerative disorders: preclinical gains and issues for translation into clinical studies. PMID- 16374322 TI - Ciliary neurotrophic factor therapy for inherited retinal diseases: pros and cons. PMID- 16374323 TI - Surmountable challenges in translating pigment epithelium-derived factor (PEDF) therapy from animal models to clinical trials for retinal degenerations. PMID- 16374324 TI - Inhibition of poly(adenosine diphosphate-ribose) polymerase (PARP) in experimental models of neurologic diseases: cell death prevention. PMID- 16374325 TI - T-cell-based vaccination against neurodegeneration: a new therapeutic approach. PMID- 16374326 TI - Animal models as tools for screening candidate drugs. PMID- 16374327 TI - Saving cone cells in hereditary rod diseases: a possible role for rod-derived cone viability factor (RdCVF) therapy. PMID- 16374328 TI - Inherited and orphan retinal diseases: phenotypes, genotypes, and probable treatment groups. PMID- 16374329 TI - Usher syndrome type 1: genotype-phenotype relationships. PMID- 16374330 TI - Retinal researchers have reasons to be optimistic. PMID- 16374331 TI - Drug delivery systems for treating orphan retinal diseases. PMID- 16374332 TI - Challenges in the pursuit of therapeutic product development. PMID- 16374333 TI - Strategies for delivery of rod-derived cone viability factor. PMID- 16374334 TI - Adenovector pigment epithelium-derived factor (AdPEDF) delivery for wet age related macular degeneration. PMID- 16374335 TI - Clinical trials with micronutrients and mineral supplements for retinal degenerations: a summary of a breakout session. PMID- 16374336 TI - A randomized placebo-controlled clinical trial of docosahexaenoic acid (DHA) supplementation for X-linked retinitis pigmentosa. PMID- 16374337 TI - New paradigms for drug discovery. PMID- 16374338 TI - Screening existing drugs for neurodegeneration: The National Institute of Neurologic Disorders and Stroke (NINDS) model. PMID- 16374339 TI - High-throughput compound screening and discovery in an academic setting. PMID- 16374341 TI - The role of RPE65 in inherited retinal diseases. PMID- 16374340 TI - The consortium project to treat RPE65 deficiency in humans. PMID- 16374342 TI - Cooperation between private and public sectors leads to an intraocular retinal implant. PMID- 16374343 TI - Preclinical assessment programs to evaluate potential therapies for the treatment of orphan retinal diseases. PMID- 16374344 TI - A network of patients with orphan retinal diseases for clinical trials: goals, structure, challenges. PMID- 16374345 TI - National and international patient networks: standardization of phenotype and genotype definitions. PMID- 16374346 TI - Design of phase III clinical trials for treatments of orphan retinal diseases: an overview of considerations. PMID- 16374347 TI - Challenges in genetic testing for clinical trials of inherited and orphan retinal diseases. PMID- 16374348 TI - Importance of genotyping in clinical trials of inherited and orphan retinal diseases. PMID- 16374349 TI - Failures and successes of clinical trials for Parkinson disease treatments. PMID- 16374350 TI - Clinical trials of neuroprotective agents in glaucoma. PMID- 16374351 TI - The national eye institute: translational clinical research initiatives on inherited and orphan retinal diseases: personal observations. PMID- 16374352 TI - Alzheimer disease: therapeutic targets for clinical trials. PMID- 16374353 TI - Governmental, university, pharmaceutical, and foundation partnerships to advance translational research in retinal disease. PMID- 16374354 TI - Fostering partnerships: a perspective from the National Institute of Neurologic Disorders and Stroke (NINDS). PMID- 16374355 TI - Fostering partnerships: The National Institutes of Health (NIH) Mission. PMID- 16374356 TI - Fostering research partnerships: a perspective from The Office of Rare Diseases (ORD). PMID- 16374357 TI - Fostering partnerships: an industry perspective. PMID- 16374358 TI - Federal regulatory issues: investigational new drugs in ophthalmology. PMID- 16374359 TI - The Food and Drug Administration's Office of Orphan Products Development: incentives, grants, and special designations speed therapies for orphan diseases. PMID- 16374360 TI - Intellectual property: patents and transfer agreements preceding clinical trials and commercialization. PMID- 16374361 TI - Intellectual property in drug development: a report from a breakout session. PMID- 16374362 TI - Mechanics of the Food and Drug Administration's Form 1571: investigational new drug application. PMID- 16374363 TI - Licensing Compounds: lessons from ISTA Pharmaceuticals. PMID- 16374364 TI - Proving inventorship: the importance of the inventor/laboratory notebook. PMID- 16374365 TI - Preclinical models of shock and sepsis: what can they tell us? AB - The goal of translational research is to transform biologic knowledge into new treatments for human disease. Although preclinical models replicate some of the features of the disease process modeled, they invariably fail to reproduce the complexity of human illness, and by their very experimental nature, they are readily manipulated to maximize evidence of efficacy. The result is that successful translation from preclinical models to clinically effective therapy is uncommon, and that clinical trials are often undertaken without a comprehensive and realistic preclinical portfolio of studies to optimize their design. The lethal and morbid human conditions of sepsis and shock are attractive targets for new therapies and enormously challenging processes to translate because they entail considerable clinical heterogeneity, require emergent effective intervention, and are shaped not only by the initial insult, but by approaches to subsequent resuscitation and support. A colloquium jointly sponsored by the Shock Society and the International Sepsis Forum in June 2004 addressed the challenges of translational research in shock and sepsis. Through a comprehensive review of a broad variety of model approaches, and vigorous debate about the merits of differing strategies, a series of common themes emerged. We concluded that there is no single ideal model of shock or sepsis, but rather a large number of complementary models that recapitulate some discrete features of the disorders while minimizing others. Consequently, successful preclinical investigation mandates the use of a panel of preclinical studies consciously designed to address specific questions of relevance to the clinical setting. A corollary of this conclusion is that preclinical studies can shape concepts of disease and can be used to refine decisions regarding optimal patient populations for therapeutic interventional trials. We further recognized that the design and reporting of preclinical studies is highly variable, thereby limiting effective data interpretation and integration between studies. Hence, greater model standardization would aid in interpreting data and in pooling results into systematic data syntheses: such efforts should be promoted and undertaken. PMID- 16374366 TI - Evaluation of endotoxin models for the study of sepsis. AB - Many strategies have been proposed for the treatment of sepsis, and most of the proposed treatment modalities have failed in clinical trials. Many of the previous treatment protocols called for blocking the activity of a single, clearly defined mediator. The underlying hypothesis was that sepsis induced a specific mediator that then caused organ injury and death. This simple, linear reasoning was frequently based on cytokines that were defined using endotoxin models of sepsis. The endotoxin models were widely used to study the pathophysiology of sepsis and were felt to adequately reproduce the full spectrum of inflammatory changes observed in patients with sepsis. Based on mortality and hematologic changes, these assumptions appeared justified. As the models were examined more closely, and directly compared with focus of infection models that more accurately portray the changes in sepsis, it became apparent that the endotoxin models did not accurately mimic the patient with sepsis. In the endotoxin models, the explosive release of cytokines into the circulating blood volume was reproducibly found regardless of the species studied (human, primate, pig, rat, or mouse). This lead to a series of anticytokine sepsis trials, all of which failed. The cytokine response in focus of infection models, such as that induced by cecal ligation and puncture, was examined and found to be more similar to that observed in patients with sepsis. When cytokine inhibitor strategies were used in the cecal ligation and puncture model, they were also generally found to lack efficacy. Compounds that have been shown to be effective at reducing mortality in endotoxin models should be re-evaluated in more clinically relevant models of sepsis. PMID- 16374367 TI - Pneumonia models and innate immunity to respiratory bacterial pathogens. AB - Preclinical sepsis models have been used for decades to study the pathophysiologic processes during sepsis and shock. Although these studies revealed promising immunomodulating agents for the treatment of sepsis, clinical trials evaluating the efficacy of these new agents in patients with sepsis were disappointing. The main reason for this unsatisfactory experience might be that unlike the clinical situation, most of these preclinical models are devoid of a localized infectious source from which the infection disseminates. Studies on the effects of several immunomodulating strategies have demonstrated strikingly opposite results when sepsis models with a more natural route of infection, such as pneumonia, were used. In this review, we will give insights into pneumonia models and discuss results and differences in the innate immune responses during distinct pulmonary infection models. PMID- 16374368 TI - Rodent models of intra-abdominal infection. AB - The first question to ask when deciding which model to use for the preclinical testing of a therapeutic agent should be: What exactly is it that the chosen model is attempting to model? Therefore, in the context of intra-abdominal models of infection, the question becomes, is the goal to mimic diffuse peritonitis, intra-abdominal abscesses, septic shock, or a multiple organ dysfunction-type syndrome. Having decided on the clinical situation to model, it becomes important to ensure that the model is as congruent with the clinical situation as feasible, especially when the goal is the preclinical testing of possible therapeutic agents. Consequently, different types of rodent intra-abdominal infection models will be reviewed, focusing on their rationales as well as their strengths and weaknesses as models of clinical disease. PMID- 16374369 TI - Preclinical models of traumatic, hemorrhagic shock. PMID- 16374370 TI - Shock and hemorrhage: an overview of animal models. AB - Shock resulting from life-threatening blood loss (hemorrhage) remains a common complication of traumatic injury. Intensive experimental efforts are needed if we are to understand the pathological effect(s) of hemorrhagic shock, alone or in association with traumatic tissue injury, and to reverse this deleterious process in trauma patients. Here, we overview selected studies that are representative of the different hemorrhagic shock models, considering their advantages and disadvantages from a scientific and clinical perspective. Fixed-pressure versus fixed-volume versus uncontrolled hemorrhage models, with or without tissue injury, will be discussed, as well as small versus large animal models. Most of these models are nonlethal in nature, and allow the researcher to understand the changes that contribute to increased susceptibility to subsequent infection or the development of multiple organ failure. We also consider some of the confounders in these models, including anesthesia, the nature of resuscitation, and the use of anticoagulants. The selection of model must take into consideration not only the need for experimental control but must also adequately reflect the clinical pathobiology of shock if we are to develop better pharmacological interventions. PMID- 16374371 TI - Inflammatory/cardiovascular-metabolic responses in a rat model of burn injury with superimposed infection. AB - Infection remains the major cause of morbidity and mortality in burn patients. Furthermore, the use of antibiotics in such patients has led to the prevalence of antibiotic-resistant microbial infections; one such infection in intensive care unit turns out to be caused by the enterococcal organisms. Our laboratory studies have used a rat model of bum injury and Enterococcus faecalis infection. Sprague Dawley male rats ( approximately 250 g) were initially given an intragastric gavage of the antibiotic ciprofloxacin for 3 days. This procedure allowed for decontamination of intestine of gram-negative and some gram-positive organisms. The remainders of the gram-positive organisms were, to a large extent, Enterococci. After the decontamination procedure, rats were intra-abdominally inoculated with E. faecalis; inoculation involved preparation of sterilized rat fecal pellets impregnated with E. faecalis (10 colony-forming units) and their implants through a midline abdominal wall incision. Some of the rats that were implanted with the fecal pellets were subject to full-thickness skin bums ( approximately 30% total body surface area; 95 degrees C water for 7 s). Sham abdominal infection rats received a sterile pellet only; sham bum procedure consisted of exposing the skin to room temperature water. All sham and bum and/or E. faecalis infection procedures were carried out on rats under pentobarbital anesthesia. Inflammation and innate host defense-related responses were assessed via measurements of neutrophil effector responses, i.e., oxygen anion free radical (O2)/eIastase production, CD11b/CD18 expression, apoptosis, and tissue infiltration. Determining epithelial lactulose permeability, microvascular albumin leakage, and epithelial tight junction integrity assessed the status of intestinal function/structural derangements. The animals' metabolic and cardiovascular integrity was evaluated determining blood pH, p02, pC02, heart rate, respiratory rate, blood pressure, and cardiac output. Whereas the aforementioned measurements were carried out at 24 to 48 h postbum injury with and without the Enterococcal infection, animal mortality was determined for up to 5 days after the experimental injuries. The results of the studies indicated that whereas bum or E. faecalis infection alone did not produce significant mortality, the dual insult with bum and E. faecalis infection resulted in significant animal death accompanied by relatively more profound metabolic and cardiovascular derangements. Inappropriately heightened neutrophil effector responses were present with bum alone as well as with the dual bum and infection complications. These studies suggest that animal models of bum injury with Enterococcal infection complications simulate the adverse outcomes bum patients infected with Enterococcal organisms. PMID- 16374372 TI - Acute pancreatitis: models, markers, and mediators. AB - Acute pancreatitis has an incidence of approximately 40 cases per year per 100,000 adults. Although usually self-limiting, 10% to 20% of afflicted patients will progress to severe pancreatitis. The mortality rate among patients with severe pancreatitis may approach 30% when they progress to multisystem organ failure. The development of acute pancreatitis illustrates the requirement for understanding the basic mechanisms of disease progression to drive the exploration of therapeutic options. The pathogenesis of acute pancreatitis involves the interplay of local and systemic immune responses that are often difficult to characterize, particularly when results from animal models are used as a foundation for human trials. Experimental studies suggest that the prognosis for acute pancreatitis depends upon the degree of pancreatic necrosis and the intensity of multisystem organ failure generated by the systemic inflammatory response. This suggests an intricate balance between localized tissue damage with proinflammatory cytokine production and a systemic, anti-inflammatory response that restricts the inappropriate movement of proinflammatory agents into the circulation. The critical players of this interaction include the proinflammatory cytokines IL-1beta, TNF-alpha, IL-6, IL-8, and platelet activating factor (PAF). The anti-inflammatory cytokines IL-10, as well as TNF-soluble receptors and IL-1 receptor antagonist, have also been shown to be intimately involved in the inflammatory response to acute pancreatitis. Other compounds implicated in disease pathogenesis in experimental models include complement, bradykinin, nitric oxide, reactive oxygen intermediates, substance P, and higher polyamines. Several of these mediators have been documented to be present at increased concentrations in the plasma of patients with severe, acute pancreatitis. Preclinical work has shown that some of these mediators are markers for disease activity, whereas other inflammatory components may actually drive the disease process as important mediators. Implication of such mediators suggests that interruption or blunting of an inappropriate immune response has the potential to improve outcome. Although the manipulations of specific mediators in animal models may be promising, they may not transition well to the human clinical setting. However, continued reliance on experimental animal models of acute pancreatitis may be necessary to determine the underlying causes of disease. Full understanding of these basic mechanisms involves determining not only which mediators are present, but also closely documenting the kinetics of their appearance. Measurement of the inflammatory response may also serve to identify diagnostic markers for the presence of acute pancreatitis and provide insight into prognosis. Understanding the models, documenting the markers, and deciphering the mediators have the potential to improve treatment of acute pancreatitis. PMID- 16374373 TI - Cecal ligation and puncture. AB - The model of cecal ligation and puncture (CLP) in rodents has been used extensively to investigate the clinical settings of sepsis and septic shock. This model produces a hyperdynamic, hypermetabolic state that can lead to a hypodynamic, hypometabolic stage, and eventual death. Blood cultures are positive for enteric organisms very early after CLP. The model has been widely used over the past 26 years and is highly versatile in adapting to a range of severity and testing objectives. It is inexpensive to prepare and technically straightforward. Aspects of sepsis research investigated using CLP include energetics, metabolism, resuscitation, antibiotic therapy, microbial factors, cardiovascular responses, immune function, mediator release, and cytokine expression patterns. The challenge of the small circulating blood volume in rodents can be overcome by using micromethods that enable analysis of small volumes, or alternatively, by using a large number of animals to obtain serial samples. PMID- 16374374 TI - Mouse models of resuscitated shock. AB - Studies of sepsis in humans are difficult because the seriousness of the disease mandates immediate intervention and because the heterogeneity of patient presentations imposes substantial limitations on clinical trials. Thus, animal models have been used extensively to explore the pathogenesis of sepsis and to generate preclinical data for therapeutic interventions. Translation of findings in these models into therapeutic strategies has been difficult, in part because of limitations in preclinical models and in part to imperfect understanding of the pathophysiology of sepsis. It is important to use an animal model that reproduces the relevant physiologic parameters present in patients with septic shock. Mouse models are particularly useful for the dissection of molecular mechanisms of disease because of the proliferation of transgenic strains. We have developed a murine model of sepsis with fluid resuscitation and antibiotic treatment that reproduces the timing and degree of mortality seen in patients with septic shock. Using continuous micromanometric pressure monitoring and assessment of hemodynamics by echocardiography, we have shown that this model reproduces the hyperdynamic state with hypotension seen in clinical sepsis. The use of transgenic technology in appropriate murine models is exciting because of its potential to permit significant strides in our understanding of the molecular mechanisms of sepsis, multiple organ system failure, and other diseases. The use of reproducible and clinically relevant mouse models of shock is essential for delineation of pathogenetic mechanisms and for initial testing of potential therapeutic strategies. PMID- 16374375 TI - The use of immunocompromised animals as models for human septic shock. AB - Although it is generally understood that no single animal model truly reflects human sepsis, the study of sepsis in immunocompromised animals is highly relevant to human sepsis research. The majority of patients with severe sepsis have significant underlying diseases that may alter innate immune defenses, disrupt microbial clearance mechanisms, and complicate the pathophysiology of human sepsis. Septic shock itself has significant effects upon the innate and adaptive host immune responses that may contribute to a state of sepsis-induced immune dysregulation. A number of animal models of sepsis displaying an array of immunocompromised states are now available. Most of these systems are small animal models with genetically defined defects of immune defenses or acquired defects from receipt of immunosuppressive or myeloablative agents. Greater emphasis should be placed on preclinical models of immunocompromised animals in the future to assess the potential clinical utility of novel drugs for human septic shock. PMID- 16374376 TI - The two-event construct of postinjury multiple organ failure. PMID- 16374377 TI - Porcine models of severe sepsis: emphasis on porcine peritonitis. PMID- 16374378 TI - Inhalational and acute lung injury. PMID- 16374379 TI - Large animal models: baboons for trauma, shock, and sepsis studies. AB - A few limited examples of large animal models are outlined, with the main emphasis on baboon models. The baboon offers all the advantages of a large animal and is comparable with humans in nearly all physiological and immunological aspects. In addition, cross-reactivity with human therapeutic and diagnostic reagents allows testing of new species-specific therapies such as antihuman antibodies, on the one hand, and monitoring with available human analytical procedures, on the other. PMID- 16374380 TI - Human endotoxemia: a model for mechanistic insight and therapeutic targeting. AB - The diversity of phenotypic manifestations, comorbidities, and therapeutic algorithms in patients with severe inflammation have confounded efforts to translate mechanistic insights from the bench top to the bedside. This dilemma has negatively impacted upon many therapeutic interventions that exhibited seemingly well-reasoned preclinical portfolios. Prudence urges the assessment of potent immunoregulatory therapies, wherever possible, in models that replicate the clinical phenotype absent overt manifestations of genetically or environmentally modified processes. The healthy human model of endotoxin administration (systemic or endobronchial) provides such an opportunity and has been used to great advantage for gaining insight into mechanisms of disease and for determination of therapeutic signal strength. When thoughtfully interpreted, the model may provide proof of principle as well as lessen the unpredictability of clinical responses. Although the broad characteristics of this model are well described in the literature, it is recognized that this model does not fully replicate the magnitude of initial inflammatory stress nor the latent spectrum of inflammation/sepsis-inducible organ system pathologies. Nevertheless, the similarities between the early, transient clinical phenotype, inducible physiochemical change, and biochemical pathway activation of this model to the early hyperdynamic phase of resuscitated injury and infection are striking. Rational testing of a therapeutic mechanism requires a quantifiable and reproducibly altered marker of the hypothetical mechanism. Given the modest nature of endotoxin induced insult, interventions that demonstrate target specific efficacy in conjunction with attenuated phenotype responses are more likely to exhibit efficacy within lower risk patient populations. By contrast, the model cannot predict clinical efficacy among higher risk patients nor in those who have endured extended periods of inflammatory stress. PMID- 16374381 TI - Gender differences in acute response to trauma-hemorrhage. AB - To understand the pathogenesis of a disease, experimental models are needed. A good experimental model is the one that simulates responses observed in the clinical setting. In recent years, clinical studies have indicated that gender might be a factor that plays a significant role in the outcome of patients with shock, trauma, and sepsis. These observations are now being evaluated in experimental setting. Studies performed in a rodent model of trauma-hemorrhage have concluded that alterations in immune and cardiac functions after trauma hemorrhage are more markedly depressed in adult males, and ovariectomized and aged females. However, both are maintained in castrated males and in proestrus females. Moreover, the survival rate of proestrus females subjected to sepsis after trauma-hemorrhage is significantly higher than age-matched males or ovariectomized females. Although these observations suggest gender-specific response after trauma-hemorrhage, the mechanisms responsible for gender specificity remain largely unknown. Furthermore, in other injuries such as burn, experimental studies dealing with sexual dimorphism are limited. Therefore, more studies in clinical and experimental settings are required to determine whether gender-specific responses are global across the injuries or are observed in specific injury situations. Studies are also needed to delineate underlying mechanisms responsible for differences between males and females after trauma hemorrhage. The information gained from the experimental studies will help in designing innovative therapeutic approaches for the treatment of trauma patients. PMID- 16374382 TI - Neutralization of tumor necrosis factor in preclinical models of sepsis. AB - Tumor necrosis factor (TNFalpha), a cardinal early mediator of the innate host inflammatory response, has been an attractive target for therapeutic intervention in human sepsis. However, pooled data from 12 completed randomized controlled trials show only a very modest impact on mortality in a highly heterogeneous population of patients. To gain insight into the preclinical in vivo biology of TNFalpha that might aid in better identifying appropriate patient populations for therapeutic intervention, we undertook a systematic review of published reports of preclinical studies assessing the consequences of neutralization of TNFalpha in models of acute infection or inflammation. We identified 143 reports incorporating 484 unique experimental comparisons in seven different animal species. The effects of neutralization of TNFalpha in these were quite variable. Neutralization of TNFalpha was beneficial in endotoxemia, or after systemic challenge with gram-negative organisms, Staphylococcus aureus, or Group B streptococci. On the other hand, neutralization was detrimental in infections caused by Streptococcus pneumoniae, Candida spp., or intracellular pathogens such as Listeria and Mycobacterium tuberculosis, and in models of pneumonia. Treatment was more efficacious when delivered before infectious challenge, and the therapeutic signal increased as the baseline mortality in the placebo group increased. Evidence of neutralization of TNFalpha bioactivity, and of attenuation of inflammation, was typically accompanied by evidence of impairment of antimicrobial defenses. Multiple specific and nonspecific therapeutic strategies were identified. We conclude that the beneficial effects of TNF in systemic inflammation occur at the cost of impaired antimicrobial defenses, and that a better understanding of the consequences of neutralization of TNFalpha in vivo could aid in better defining optimal patient populations for therapeutic intervention. PMID- 16374383 TI - The effects of granulocyte colony-stimulating factor in preclinical models of infection and acute inflammation. AB - The cytokine granulocyte colony-stimulating factor (G-CSF) is a potent endogenous trigger for the release of neutrophils from bone marrow stores and for their activation for enhanced antimicrobial activity. G-CSF has been widely evaluated in preclinical models of acute illness, with generally promising though divergent results. A recombinant G-CSF molecule has recently undergone clinical trials to assess its efficacy as an adjuvant therapy in community-acquired and nosocomial pneumonia, however, these studies failed to provide convincing evidence of benefit. We undertook a systematic review of the published literature reporting the effects of modulation of G-CSF in preclinical in vivo models to determine whether evidence of differential efficacy might explain the disappointing results of human studies and point to disease states that might be more likely to benefit from G-CSF therapy. G-CSF has been evaluated in 86 such studies involving a variety of different models. The strongest evidence of benefit was seen in studies involving intraperitoneal challenge with live organisms; benefit was evident whether the agent was given before or after challenge. G-CSF demonstrates anti-inflammatory activity in models of systemic challenge with viable organisms or endotoxin, but only when the agent is given before challenge; evidence of benefit after challenge was minimal. Preclinical models of intrapulmonary challenge only show efficacy when the cytokine is administered before the infectious challenge, and suggested harm in gram-negative pneumonia resulting from challenge with Escherichia coli or Klebsiella. There is little evidence for therapeutic efficacy in noninfectious models of acute illness. We conclude that the most promising populations for evaluation of G-CSF are neutropenic patients with invasive infection and patients with intra-abdominal infection, particularly those with the syndrome of tertiary, or recurrent, peritonitis. Significant variability in the design and reporting of studies of preclinical models of acute illness precludes more sophisticated data synthesis. PMID- 16374386 TI - [Proper antibiotic use]. PMID- 16374387 TI - [Professional associations in the evaluation of professional competence]. PMID- 16374388 TI - [Appropriate use of antibiotics in hospitals. Investigating clinical practices in management of community-acquired pneumonia]. AB - INTRODUCTION: The rationalization of medical practices for antibiotic use in hospitals is necessary to improve both the cost-efficiency and effectiveness of health care. This study sought to investigate the impact of implementation of local management guidelines for inpatient community-acquired pneumonia (CAP). METHODS: This retrospective, comparative study measured the quality of antibiotic prescriptions with the 10-item Medication Appropriateness Index (MAI). Clinical and demographic characteristics, as well as process-of-care and outcome indicators, were recorded for all patients with CAP admitted to a medical ward at the Nantes university hospital during two 12-month periods: before (Period A, 39 patients) and after (Period B, 50 patients) implementation of local guidelines. RESULTS: The MAI was significantly higher during period B than period A (5.1 points compared with 2.2, p=0.0001). Guideline implementation shortened the mean duration of antibiotic treatment (13.1 versus 16.0 days, p=0.0003) and of intravenous treatment (3.4 versus 4.7 days, p=0.04). The mean duration of hospital stay also fell substantially (7.4 versus 15.0 days, p=0.0001), as did the mean cost of antibiotics (35.4 versus 64.1 euros, p=0.003). DISCUSSION: The MAI confirmed that antimicrobial practices varied significantly and that appropriate local guidelines improved the quality of antibiotic prescriptions and saved hospitalization costs. CONCLUSION: The MAI, which assesses the appropriateness of antibiotic use in hospitals, could be an interesting tool for prospective use as an indicator of quality-of-care improvement and for more efficient use of available health care resources. PMID- 16374389 TI - [Local guidelines and quality of antibiotic treatment in urinary tract infections: a clinical audit in two departments of a university hospital]. AB - AIM: To assess the effect of local guidelines implemented at the Nantes University Hospital regarding antibiotic therapy for urinary tract infections. DESIGN: Before/after study of one medical ward and one urologic surgery ward. Quality was measured by two principal criteria: compliance with guidelines and medical justification in the specific clinical situation. Both criteria considered simultaneously the choice of drug, dose and duration of treatment. Deviations from the guidelines were described. RESULTS: 1086 UTI cases were identified over two 12-month periods, before and after the dissemination of guidelines (for prostatitis, pyelonephritis, indwelling catheter-associated UTIs, and other undefined UTIs). The guidelines were applicable in 313 (30%) cases. Overall, after implementation of the guidelines, the percentage of justified prescriptions did not change significantly (41.8% compared with 38.7%, p=0.299), but the percentage of correct (conforming) prescriptions fell (from 30.4% to 15.7%, p=0.0022). The percentages of correct and justified prescriptions differed in the medical (respectively 45.0% and 46.6%,) and surgical units (13.1% and 36.5%). CONCLUSIONS: Issuing guidelines does not necessarily improve the quality of antibiotic therapy for UTIs in hospitals. PMID- 16374390 TI - [Defining and assessing competence of healthcare professionals. A survey of 148 organizations]. AB - INTRODUCTION: To study how competence is described and perceived by organizations of healthcare professionals in France and propose approaches toward implementing a policy to monitor and assess competence throughout medical professionals' careers. METHODS: After a literature search, we sent 160 letters to organizations representing 16 healthcare professions (listed in the French healthcare code) describing our objectives and proposing interviews. Semi-structured interviews (45-90 minutes) were conducted. The principle questions asked were: What is your organization's definition of competence? What are the principal elements that define competence in healthcare activities? How can a system for assessing competence be implemented? Which methods for such a system are most appropriate, based on experience in other countries? Who are the players in the field of competence? How can organizations participate in monitoring competence? RESULTS: 265 people representing 148 French organizations were interviewed. Analysis of the interviews showed that the principal points mentioned included: need for recognition; lack of training on new developments; need to anticipate changes; need for better health security in the healthcare system. There was a general consensus on the basic elements of competence, the responsibility of public institutions and professional organizations, and the need to work together. DISCUSSION: We suggest that competence in health care should be defined as follows: "professional competence is based on an initial diploma, participation in effective continuing medical education, a minimum amount of professional activity, and a regular peer review process". Healthcare professionals in France would like to have a better system that allows them to exchange more information on the principal issues in health care. PMID- 16374391 TI - [Spontaneous skin necrosis from acquired protein S deficiency in a renal transplant recipient]. AB - INTRODUCTION: Spontaneous skin necrosis revealed acquired protein S deficiency due to isotype G autoantibodies. CASE: This 31-year-old male renal transplant recipient, receiving immunosuppressive treatment, was hospitalized for necrotic purpural lesions. We were not able to detect any triggering factor. Sustained anticoagulant therapy remained essential to prevent new skin lesions and perhaps more thrombotic events. COMMENTS: This condition is rare in adulthood, but is well described in children's purpura fulminans, especially the post-varicella form. Its mechanism remains unclear. PMID- 16374392 TI - [Pulmonary eosinophilia in France: possible toxocariasis]. AB - INTRODUCTION: In France, except in the overseas departments and territories, pulmonary eosinophilia rarely has a parasitic cause except among subjects who have traveled to tropic areas. CASE: A 19-year-old man was hospitalized for incidentally discovered hypereosinophilia. The thoracic CT scan showed several hyperdense nodules that suggested bilateral interstitial alveolar lesions. Diagnosis was finally based on serologic results positive for toxocariasis. A 14 day course of albendazole led to regression of the eosinophilia and of the radiographic images. DISCUSSION: The frequency of toxocariasis or visceral larva migrans syndrome is probably underestimated. In adults, the symptoms can be atypical and possibly serious. Albendazole, for 14 days, is the reference treatment. PMID- 16374393 TI - [Toxocariasis mimicking liver tumor]. AB - INTRODUCTION: Toxocariasis is a common helminth infection that causes visceral larva migrans in humans. It has a wide and varied clinical spectrum. CASE: A 60 year-old woman consulted after 6 months of recurrent biliary pain with fever. Laboratory tests showed moderate disturbance of liver enzymes and hypereosinophilia (10 400/mm3). Imaging studies revealed a pseudotumor in hepatic segment IV, hyperechoic in abdominal ultrasound and low density in computed tomography. The diagnostic conclusion after ultrasound-guided liver biopsy was unspecific granulomatous hepatitis and eosinophilic infiltrate. The course of the antibody titer to Toxocara canis finally resulted in a diagnosis of Toxocara infection. Outcome was good following treatment with diethylcarbamazine. DISCUSSION: Some clinical features of liver toxocariasis can mimic tumors and may be interpreted histologically as granulomatous hepatitis, eosinophilic infiltrate of the hepatic portal vein, and/or necrotizing eosinophilic abscesses. Treatment is not yet well codified, but the symptomatic forms call for albendazole or diethylcarbamazine. PMID- 16374394 TI - [Spontaneous splenic hematoma: trauma injury or drug reaction or both?]. AB - INTRODUCTION: Spontaneous splenic hematomas are uncommon and frequently associated with infectious, hematologic, or neoplastic diseases. Presentation is typically acute but progressive forms have been described. CASE: We report the case of a 45-year-old man consulting for abdominal pain that was found to be due to a spontaneous splenic hematoma. No recent injuries, infections, or hematologic, neoplastic or gastrointestinal diseases were found, but the patient had had a minor injury 9 months earlier and had been treated with selective serotonin reuptake inhibitors (venlafaxine) for the past year. This history suggested that the drug might play a role. DISCUSSION: Although the likely cause of this splenic hematoma appears to be a minor injury 9 months before the onset of pain, we cannot rule out the possibility that selective serotonin reuptake inhibitor treatment was a predisposing factor. PMID- 16374395 TI - [Prenatal treatment of congenital toxoplasmosis]. PMID- 16374396 TI - [Hereditary hypophosphatemia in adults]. AB - Hereditary hypophosphatemic rickets groups together X-linked hypophosphatemic rickets (XLH), autosomal dominant hypophosphatemic rickets (ADHR) and hereditary hypophosphatemic rickets with hypercalciuria (HHRH, autosomal recessive). Clinical and biological characteristics and treatment depend on specific etiology. Mutations causing hereditary hypophosphatemic rickets involve PHEX located on Xp11.22 for XLH and FGF-23 located on 12p13 for ADHR. The gene involved in HHRH remains unknown: candidates may encode proteins that modulate phosphate transporter expression or activity. Others forms of rickets must be ruled out: acquired hypophosphatemia due to oncogenic osteomalacia, X-linked recessive hypophosphatemic rickets or Dent's disease, and hereditary 1, 25 dihydroxyvitamin D-resistant rickets with a defect either in the 1-alpha hydroxylase gene (pseudo-vitamin D deficiency rickets, PDDR) or in the vitamin D receptor (hereditary vitamin D-resistant rickets, HVDRR). PMID- 16374398 TI - [Dudley Joy Morton's foot syndrome]. AB - Morton's foot syndrome is a hereditary syndrome characterized by a short first metatarsal bone, posterior displacement of the sesamoids, and hypertrophy of the second metatarsal, causing excessive weight to be borne by the second metatarsal head. This condition results in callus formation under the second metatarsal. Pain and tenderness are usually felt at the base of the first two metatarsal bones and at the head of the second. Pain (metatarsalgia) may be disabling. Conservative treatment consists of placing a flexible pad under the first metatarsal and toe to increase the range of motion and weight-bearing along the first metatarsophalangeal joint and the hallux. Surgical treatment consists of removing a small portion of bone from one or two joints to bring the toe down to the desired length. Lengthening short toes by placing a silicon implant into one of the joints is also possible. PMID- 16374397 TI - [Mechanisms and treatment of psoriasis]. AB - Psoriasis is a common dermatosis that affects 3-5% of the European population. Current treatments offer considerable clinical benefits, but their use is limited due to tolerance problems. Recent years have seen the development of new treatments, used separately or in combination to improve the chronic lesions caused by this disease. T cells play an important role in the pathogenesis of psoriasis. Various techniques target the T cells and the immunological mechanisms involved in their activation. In 2005, treatment of psoriasis is directed essentially towards immunological pathways. PMID- 16374399 TI - [Mediastinal thymolipoma]. PMID- 16374400 TI - Statins for high-risk patients without heart disease or high cholesterol. PMID- 16374401 TI - A combination of oxycodone and ibuprofen (Combunox) for pain. PMID- 16374402 TI - Drugs for treatment of heart failure. PMID- 16374403 TI - Tethered cord syndrome. AB - AIM: Tethered cord syndrome (TCS) is a stretch-induced disorder of the spinal cord. Tethering is due to an inelastic structure anchoring the caudal end of the spinal cord as a short and thick filum terminale. Spinal dysraphism is occasionally associated,but the etiological relationship between these disorders remain unclear. Other anomalies may be concurrently found as hydromielia and Arnold-Chiari malformation. METHODS: The authors analysed neuroradiological findings in 5 children and 9 patients of 20-24 years of age; there were four male and ten female. The criteria for inclusion were neurological disturbances (disorders!) localizable to the level of the conus and evidence for spinal dysraphism. The purpose of this study was to make the precise diagnosis and make also precise planning for therapy, conservative or surgical treatment. RESULTS: The age of diagnosis of TCS varies from under 1 year to 14 years and is very rarely as late as adulthood. TCS can present late and insidiously with progressive gait disturbances, atrophy of various muscle groups or the entire limb, loss of reflexes,loss of sensation in the sacral dermatomes, sphincter disturbances, gait abnormality and pain in the gluteal, perianal and other pelvic areas. The diagnosis involves standard X-RAY examination, CT and CT-Mielography but MRI is now a diagnostic method of choice. Surgical untethering of the cord is recommended. The associated pain responds best to surgical treatment; ambulation and bladder function may improve as well. CONCLUSIONS: However sphincter dysfunction often remains a permanent problem. Given the potential for rapid deterioration with incomplete neurological recovery, even prophilactic surgery seems advisable. The patient need to be followed up, if not operated upon. PMID- 16374404 TI - Posterior lumbar interbody fusion with peek cages: personal experience with 20 patients. AB - AIM: The use of interbody cages to obtain fusion in the lumbar spine has increased noticeably in the last few years. They are used both with and without posterior stabilization. In our institution a prospective analysis to determine whether PEEK cages can be used as a stand alone device has been performed. METHODS: The 20 patients with PEEK cages had clinical and radiological follow-up with controls at 1, 3, 6 and 9 months. RESULTS: The clinical results were considered satisfactory in 75% of the cases. There were no intra-or perioperative complications. There was no displacement of the cages. No signs of unsuccessful fusion were observed. CONCLUSIONS: Interbody PEEK cages fulfill the objective of stabilizing the treated segment immediately and subsequently. They can be used as stand alone devices with the correct surgical technique and following precise indications. PMID- 16374405 TI - Anatomical description of the facet joint innervation and its implication in the treatment of recurrent back pain. AB - AIM: Many techniques are used in the back pain treatment, standing out the facet denervation as a therapeutic option for pain that originates in the facet joints. It's known that the facet joint is an abundant area of nocireceptor innervation, although the distribution and the location of the involved branches have not being well demonstrated. A good comprehension about the affected innervation is very important to get an effective treatment. Purpose of study was to describe innervation of the lumbar facet joints, potentially used in the diagnosis and treatment of painful pictures of the lumbar region by facet syndrome. STUDY DESIGN: anatomical study of nerve roots distribution of the facet joint 3 human corpses. The determination of the neurotomy s point was carried out by direct visualization and the radiological study in human parts. METHODS: Three anatomical pieces of the human lumbar spine were dissected. In those 3 pieces, the facet joint innervation distribution was studied thoroughly using surgical microscope and microsurgical technique. In one of the pieces the needles positioning was first made to test through the radiological study the possible application of the precise denervation in low back pain treatment. RESULTS: The L1 to L4 segments, each dorsal branch of root emits a medial branch that emerges from intertransversal ligament. This branch crosses the superior margin of the medial termination of transverse process, passing through the root of the superior articulate process. Each branch innerves the anterior region of the inferior facet and the inferior portion of articulation which one spins around. The L5 dorsal branch was larger than the superior branches. It emerges dorsally and in the inferior region on top of the sacrum wing. This nerve is in the bone fissure of the junction between the wing and the posterior region of the sacrum articular process. Near the inferior portion of the articular process, the nerve ramifies itself in lateral and medial branch. The medial branch comes back around the inferior portion of the lumbar-sacrum articulation that it innervates. CONCLUSIONS: We didn't note great variations in the anatomy from L1 do L4. The L5 segment has a different distribution of the branches that should be considered when we do a percutaneous denervation procedure. The approach of the needle must touch the transverse process and feels the resistance of the articular joint . The determination of the neurotomy s point tends to become more precise denervation procedure. PMID- 16374406 TI - Balloon kyphoplasty in traumatic fractures of the thoracolumbar junction. Preliminary experience in 12 cases. AB - AIM: The purpose of this study was to assess the effectiveness and safety of Ballon Kyphoplasty as a new method of treatment for traumatic fracture of the thoracolumbar junction. METHODS: We treated twelve patients 8 patients male and 4 female. The average age was 47,3 years (22-75 years). The interested vertebrae and therefore the procedures have been 13 since in a patient two vertebrae have been treated. In 9 cases the interested vertebra was L1, in 2 cases L2 and one case D11 and D12. Following the classification of Magerl 9 fractures were of A1.2 fractures and one fracture respectively of type A1.3, A2.2, A2.3, A3.1. We have never placed indication to Balloon Kyphoplasty in the fractures of the type B and C, neither in the complete burst fractures (A3.2 and A3.3), neither in the fractures of the A1.1 type. Balloon Kyphoplasty has been carried out using the Kyphon system (Kyphon Inc. Sunnyvale, CA, USA). In the patients older than 50 years (7 cases) we have used polymethylmethacrylate (PMMA), as filling material, while in the patients younger than 50 years (5 in total), we have used Calcibon (Biomet, Inc.Warsaw, Indiana) in 4 cases and in one case a new material called KyphOs (Kyphon Inc. Sunnyvale, CA, USA). These last two types of material are made up of tricalcium phosphate that, although less manageable of the PMMA and less resistant initially, in the time it will be reabsorbed and osteointegrated. RESULTS: The method demonstrated swift pain relief associated with an evident augmentation in the resistance and restoration of the vertebral body's physiological shape. No patient has worn a postoperative orthesis. In all the cases we obtained an optimal stabilization in the follow-up minimal to 4 months. In no case we observed a structural yielding of the vertebra. CONCLUSIONS: Kyphoplasty is an effective, alternative, simple and safe treatment of traumatic fracture of the thoracolumbar junction. PMID- 16374407 TI - Demonstrated rapid growth of a corpus callosum cavernous angioma within a short period of time. AB - Cavernous angiomas are uncommon central nervous system vascular malformations. They occur in the corpus callosum very rarely. In this study we report a case of corpus callosum cavernous angioma which demonstrated rapid growth within a short period of time. Corpus callosum cavernous angiomas have distinct features regarding growth and should be treated more carefully by giving more importance to surgical removal rather than a conservative approach. PMID- 16374408 TI - Cavernous hemangioma of the parietal bone. A case report. AB - Intraosseous cavernous hemangiomas are a rare finding in the calvarium. It is a benign tumors arising from the intrinsic vasculature of the bone. We report one case observed in a 31 year-old female, presented with a progressive left parietal mass. A plain X-ray of the skull and cranial CT-scan showed a osteolytic lesion in the left parietal bone. Surgery consisted of total resection of the lesion and cranioplasty. The postoperative course was uneventful. Histological study revealed a cavernous hemangioma of the diploe. In view of this observation and the literature review, clinical; radiological and therapeutic aspect of this rare entity are discussed. PMID- 16374409 TI - Sylvian subarachnoid and extra-pial cavernous angioma. AB - Cavernous angiomas are one of the four types of vascular malformations of the central nervous system. Lesions situated subarachnoidally are rare although some cases with an infratentorial localization have been reported. Our case represents an unusual localization of a subarachnoid cavernous angioma. A 35- year-old patient was admitted to our department with signs of subarachnoid hemorrhage. A 1 cm hyperdense lesion placed at the right Sylvian fissure was distinguished by CT scan examination and no arterial supply was revealed on cerebral angiography. Surgical intervention showed a lesion placed extra-pial and totally encased in the subarachnoid space in the superficial part of the Sylvian cistern. This case represents a radiologically visible supratentorial extra-pial subarachnoidal cavernous angioma. This case provides confirmation of one of the supposed causes of subarachnoid hemorrahage ''sine materia''. PMID- 16374412 TI - Nephrologists sans frontieres. PMID- 16374413 TI - Respiratory distress in a patient with rhabdomyolysis-induced acute renal failure. PMID- 16374414 TI - Leukocytes in tubulointerstitial inflammation. AB - The work of Lange-Sperandio et al in this issue explores the differential role of beta2 integrins in promoting the macrophage infiltration characteristic of the obstructed kidneys of neonatal mice. Future work is needed to define factors that regulate macrophage death within or emigration from the kidney as well as to explore strategies to modulate macrophage phenotype. This knowledge will assist the development of novel therapeutic agents to limit injury and promote tissue repair. PMID- 16374415 TI - Role of the aldosterone-sensitive distal nephron in the sodium retention associated with liver cirrhosis. AB - The renal mechanisms underlying sodium retention during liver cirrhosis have been difficult to elucidate. Kim and associates describe a biphasic pattern of regulation of the renal epithelial sodium channel in the common bile duct ligation model, shedding some light on this issue. PMID- 16374416 TI - Receptor-mediated actions of renin and prorenin. AB - Renin can induce renal disease by generating angiotensin II and, thereby, increasing fibrosis. Huang et al describe a new mechanism of action whereby the renin-angiotensin system can also exert this effect. Direct activation of the renin/prorenin receptor in mesangial cells induced synthesis of TGF-beta and profibrotic proteins. Hence, like other proteases such as thrombin, renin and prorenin are capable of receptor-mediated cellular signaling. PMID- 16374417 TI - Hypoxia-inducible factors: where, when and why? AB - Hypoxia-inducible factor (HIF) is a family of transcription factors that regulate the homeostatic response to oxygen deprivation during development, physiological adaptation, and pathological processes such as ischemia and neoplasia. Our understanding of the function of different HIF isoforms is being advanced by understanding the processes that regulate their activity, learning where and when they are expressed and what genes they regulate. PMID- 16374418 TI - Acute renal failure outcomes in children and adults. AB - Acute renal failure (ARF) in hospitalized patients remains associated with significant morbidity and mortality. Most reports have detailed the in-hospital outcomes in ARF patients. This commentary focuses on the outcomes (including quality of life) of ARF patients, both adults and children, after discharge from the hospital. PMID- 16374419 TI - 'Hypertension' and 'microalbuminuria': the bell tolls for thee. AB - Since the terms 'hypertension' and 'microalbuminuria' were first defined, data from numerous studies have documented the continuous, rather than dichotomous, relation between blood pressure, albumin excretion, and cardiovascular disease. Lower blood pressures, down to at least 115/75 mmHg, and lower albumin excretions, below an estimated 2 mg/day, are associated with less cardiovascular risk. We hypothesize that the abundances of modern civilization superimposed on the Paleolithic genotype of humans, which has not substantially changed in the last 10 000 years, have considerably shifted the 'normal' values for blood pressure and various biochemical indices such as albuminuria still found in today's stone-aged cultures to the 'neo-normal' values observed today in the rest of the modern world. Defining a large portion of the population as 'normal' based upon these dichotomous 'neo-normal' standards is not supported by the data, and therefore seems unjustifiable. We propose that the medical community consider abandoning the terms 'hypertension' and 'microalbuminuria' in favor of 'blood pressure-associated' and 'albuminuria-associated' disease. PMID- 16374420 TI - Progression of kidney disease: blocking leukocyte recruitment with chemokine receptor CCR1 antagonists. AB - Chronic kidney disease (CKD) is usually associated with interstitial leukocytic cell infiltrates, which may contribute to disease progression by production of proinflammatory, proapoptotic, and profibrotic mediators. Recruiting leukocytes into the kidney involves local expression of chemotactic cytokines, that is, chemokines, that interact with respective chemokine receptors on the leukocyte's outer surface. Thus, specific chemokine receptor antagonists may represent an attractive therapeutic concept to interfere with renal leukocyte recruitment. Among the proinflammatory chemokine receptors, chemokine receptor (CCR)-1 has nonredundant roles for leukocyte adhesion to activated vascular endothelium and for transendothelial migration. In fact, blocking CCR-1 with specific small molecule antagonists was shown to retard progression in various types of rodent CKD models. Here we discuss the perspective of CCR-1 as a new potential target for the treatment of CKD. PMID- 16374421 TI - Vitamin D in chronic kidney disease: a systemic role for selective vitamin D receptor activation. AB - Hyperparathyroidism occurs in most patients during the progression of chronic kidney disease (CKD) and one of its initiating events, reduced serum levels of 1,25-dihydroxyvitamin D, results from a decrease in renal 1alpha hydroxylase activity, which converts 25-hydroxyvitamin D to its activated form. The combination of persistently high parathyroid hormone (PTH) and low 1,25 dihydroxyvitamin D is associated with bone loss, cardiovascular disease, immune suppression and increased mortality in patients with end-stage kidney failure. Recent studies in dialysis patients suggest that paricalcitol, a selective activator of the vitamin D receptor (VDR), is associated with a more favorable efficacy to side effect profile than calcitriol, with less morbidity and better survival. One hypothesis derived from such studies suggests that systemic activation of VDRs may have direct effects on the cardiovascular system to decrease mortality in CKD. Although current guidelines for regulating serum calcium, phosphate and PTH recommend specific interventions at the various stages of CKD to prevent or postpone irreversible parathyroid disease and decrease cardiovascular morbidity and mortality, emerging data suggest that vitamin D therapy may prolong survival in this patient population by mechanisms that are independent of calcium, phosphate and PTH. It is suggested that a re-evaluation of current treatment recommendations is needed and that future research should focus on mechanisms that distinguish potential tissue specific benefits of selective VDR activators in patients with CKD. PMID- 16374422 TI - Enhanced transduction of fibroblasts in transplanted kidney with an adenovirus having an RGD motif in the HI loop. AB - Application of gene therapy to the renal graft has a powerful potential to improve the outcome of kidney transplantation and eliminate detrimental side effects associated with systemic therapy, through local expression of immunoregulatory or other protective molecules. However, the search for the optimal vector is still ongoing. In this study, we used a modified adenovirus that has an Arg-Gly-Asp (RGD) motif inserted in the HI loop of the fiber knob, as a successful strategy to transduce the renal graft. Donor Lewis rat kidneys were infused via the renal artery with a solution containing either a fiber-modified adenovirus (AdTL-RGD) or an unmodified adenovirus (AdTL), or with saline. Syngeneic recipients were killed after 3, 7 or 14 days. Efficiency, selectivity, localization, time course of gene expression and side effects were studied using biochemical and immunohistological techniques. Enhanced gene expression was achieved selectively in the transplanted kidney by AdTL-RGD, when compared to AdTL. Transgene expression lasted for at least 2 weeks. With the AdTL-RGD vector, the transgene was abundantly expressed in the renal interstitial fibroblasts. An increase in the number of cytotoxic T lymphocytes accompanied the use of either vector, when compared to saline. These data convincingly show enhanced and selective gene transfer to the fibroblasts of transplanted kidneys using an RGD modified adenovirus, providing a highly efficient vector system for future therapeutic interventions. PMID- 16374423 TI - Modulation of renal kallikrein by a high potassium diet in rats with intense proteinuria. AB - Injury of the renal tubulointerstitial compartment is recognized to play an important role in hypertension. Its damage may in turn, impair the activity of vasodepressor systems, like the kallikrein-kinin, in blood pressure regulation. The overload proteinuria model induces tubulointerstitial injury with activation of the renin-angiotensin system, but renal kallikrein and the development of hypertension have not received special attention. Sprague-Dawley rats received seven intraperitoneal doses of bovine serum albumin (BSA) 2 g/day under normosodic diet and were hydrated ad libitum. A second group received a high potassium diet to stimulate kallikrein production during the previous four weeks and while under BSA administration. A third one received potassium and BSA in the same schedule, but with the kinin B2 receptor antagonist, HOE140, added during the protein load phase. A control group received seven saline injections. Kallikrein protein was detected by immune labeling on renal sections and enzymatic activity in the urine. The BSA group showed massive proteinuria followed by intense tubulointerstitial damage. Blood pressure increased after the third dose in BSA animals, remaining elevated throughout the experiment, associated with significant reductions in renal expression and urinary activity of kallikrein, compared with controls. An inverse correlation was found between blood pressure and immunohistochemistry and urinary activity of kallikrein. Potassium induced a significant increase in both urinary activity and renal kallikrein expression, associated with significant reduction in blood pressure. The HOE140 antagonist blunted the antihypertensive effect of kallikrein stimulation in proteinuric rats. Loss of renal kallikrein, produced by tubulointerstitial injury, may participate in the pathogenesis of the hypertension observed in this model. PMID- 16374424 TI - Efficacy and immunogenicity of novel erythropoietic agents and conventional rhEPO in rats with renal insufficiency. AB - Recombinant human erythropoietin (rhEPO) is used to treat anemia in chronic renal insufficiency. Erythropoietin (EPO) immunogenicity can lead to EPO-resistant anemia. Conjugating proteins with polyethylene glycol (PEG) can prolong elimination half-life and diminish protein immunogenicity. We investigated the efficacy of new erythropoietic agents, synthesized by single (Ro 50-3821) and multiple (MIX) integrations of PEG and succinimidyl butanoic acid with rhEPO, in rats with chronic renal insufficiency. Sprague-Dawley rats with surgically induced renal insufficiency received Ro 50-3821 or MIX subcutaneously (s.c.) over 4-12 weeks compared to rhEPO and NaCl. Hemoglobin and antibody levels served as primary efficacy and safety variables. Dosing intervals and dose-response characteristics were investigated. Ro 50-3821 (2.5 microg/kg once weekly) increased hemoglobin levels by 7 g/dl after 4 weeks compared to 1 g/dl in NaCl controls (P<0.05). MIX (2.5 microg/kg once weekly) and rhEPO (0.25 microg/kg three times weekly) increased hemoglobin levels by 3 g/dl. Ro 50-3821 administered for 12 weeks (0.75 microg/kg once weekly) increased hemoglobin levels (from 13 to 19 g/dl) more effectively than rhEPO (0.75 microg/kg once weekly, decline from 13 to 11 g/dl, P<0.05). No antibodies against Ro 50-3821 were detected after 12 weeks of treatment. Antibodies against rhEPO were seen in 69% of animals (P<0.00001). Ro 50-3821 increased hemoglobin levels with once weekly s.c. dosing. Multiple pegylated EPO is less effective. In rats, rhEPO failed to increase hemoglobin levels with once weekly long-term dosing. Antibody formation following rhEPO may explain this finding. Therefore, Ro 50-3821 may provide important clinical advantages compared to unpegylated EPO. It can be administered in longer dosing intervals and has a lower risk of unfavorable immunological responses. PMID- 16374425 TI - Fructose-1,6 diphosphate as a protective agent for experimental ischemic acute renal failure. AB - Cold ischemia time is a risk factor for the development of acute renal failure in the immediate post-transplant period. In this study, we aimed to determine if intravenous fructose-1,6-diphosphate (FDP), given before nephrectomy, attenuates renal cell injury in a cold ischemia model. Male adult Wistar rats were subjected to infusion of either FDP 350 mg/kg (group F, n=6), an equal volume of 0.9% NaCl (group S, n=6), an equal volume/osmolality of mannitol (group M, n=6) or no infusion (group C, n=7). Kidneys were then perfused in situ with Collins solution and nephrectomy was performed. Other kidney slices were stored in Collins solution at 4 degrees C. Adenosine triphosphate (ATP) levels and lactate dehydrogenase (LDH) release were examined at 0, 24, 48 and 72 h. Other slices, obtained after 50 min immersion in Collins solution at 37 degrees C, were frozen for characterization of cytoskeletal preservation using phalloidin-FITC staining. Apical fluorescence intensity of proximal tubule cells, indicative of the F-actin concentration, was measured in a fluorescence microscope interfaced with computer image analysis system. Adenosine triphosphate levels, after up to 72 h of tissue incubation, were higher (P<0.05) in the FDP group when compared to other groups. In addition, LDH release was smaller (P<0.0001) in the FDP group. The F-actin concentration of proximal tubule cells cells was greater in the FDP group (P<0.0001). Results indicate that FDP is a useful tool to increase tissue viability in a rat kidney subjected to cold ischemia, by maintaining ATP cell content, decreasing LDH release and preventing microfilament disruption of proximal tubule cells. PMID- 16374426 TI - Monocyte chemoattractant protein-1 promotes the development of diabetic renal injury in streptozotocin-treated mice. AB - Diabetic nephropathy involves a renal inflammatory response induced by the diabetic milieu. Macrophages accumulate in diabetic kidneys in association with the local upregulation of monocyte chemoattractant protein-1 (MCP-1); however, the contribution of macrophages to renal injury and the importance of MCP-1 to their accrual are unclear. Therefore, we examined the progression of streptozotocin (STZ)-induced diabetic nephropathy in mice deficient in MCP-1 in order to explore the role of MCP-1-mediated macrophage accumulation in the development of diabetic kidney damage. Renal pathology was examined at 2, 8, 12 and 18 weeks after STZ treatment in MCP-1 intact (+/+) and deficient (-/-) mice with equivalent blood glucose and hemoglobin A1c levels. In MCP-1(+/+) mice, the development of diabetic nephropathy was associated with increased kidney MCP-1 production, which occurred mostly in tubules, consistent with our in vitro finding that elements of the diabetic milieu (high glucose and advanced glycation end products) directly stimulate tubular MCP-1 secretion. Diabetes of 18 weeks resulted in albuminuria and elevated plasma creatinine in MCP-1(+/+) mice, but these aspects of renal injury were largely suppressed in MCP-1(-/-) mice. Protection from nephropathy in diabetic MCP-1(-/-) mice was associated with marked reductions in glomerular and interstitial macrophage accumulation, histological damage and renal fibrosis. Diabetic MCP-1(-/-) mice also had a smaller proportion of kidney macrophages expressing markers of activation (inducible nitric oxide synthase or sialoadhesin) compared to diabetic MCP-1(+/+) mice. In conclusion, our study demonstrates that MCP-1-mediated macrophage accumulation and activation plays a critical role in the development of STZ induced mouse diabetic nephropathy. PMID- 16374428 TI - Biphasic changes of epithelial sodium channel abundance and trafficking in common bile duct ligation-induced liver cirrhosis. AB - We hypothesize that dysregulation of the epithelial sodium channel (ENaC) may be responsible for the increased sodium retention in liver cirrhosis. Liver cirrhosis was induced by common bile duct ligation (CBDL). We examined the abundance of ENaC subunits and type 2 isoform of 11beta-hydroxysteroid dehydrogenase (11betaHSD2) in the kidney by immunoblotting and immunohistochemistry at 6 or 8 weeks after operation. At 6 weeks, cirrhotic rats had developed ascites and displayed a positive sodium balance. The urinary sodium excretion and fractional excretion of sodium were decreased, while plasma aldosterone was unchanged. The abundance of ENaC subunits was not changed in the cortex and outer stripe of the outer medulla (OSOM). In contrast, immunoperoxidase microscopy revealed an increased apical targeting of alpha-, beta- and gammaENaC in late distal convoluted tubule, connecting tubule and collecting duct. Moreover, 11betaHSD2 abundance was decreased in the cortex/OSOM and inner stripe of the outer medulla. At 8 weeks, urinary sodium excretion and fractional excretion of sodium were not changed, while the plasma aldosterone level was decreased. The expression of ENaC subunits was decreased in the cortex/OSOM. Immunoperoxidase microscopy confirmed decreased expression of ENaC subunits, whereas subcellular localization was not changed. These results suggest that increased apical targeting of ENaC subunits and diminished abundance of 11betaHSD2 may contribute to promote sodium retention in the sodium-retaining stage of liver cirrhosis (at 6 weeks). The subsequent decreased expression and reduced targeting of ENaC subunits may play a role in promoting sodium excretion in the later stage of liver cirrhosis (at 8 weeks). PMID- 16374427 TI - Distinct roles of Mac-1 and its counter-receptors in neonatal obstructive nephropathy. AB - Urinary tract obstruction during renal development leads to tubular atrophy and interstitial fibrosis. Inflammatory macrophages are crucial in this process, and beta2-integrins play a major role in leukocyte recruitment. We investigated the role of beta2-integrins and their major counter-receptors (intercellular adhesion molecule-1 (ICAM-1), receptor for advanced glycation endproducts (RAGE), junctional adhesion molecule (JAM)-C) in obstructive nephropathy in neonatal mice. Two-day-old beta2-integrin-deficient mice (Mac-1-/- and LFA-1-/-(deficient for leukocyte function-associated antigen-1)) and wild-type mice (C57BL/6) underwent unilateral ureteral obstruction (UUO) or sham operation. After 1, 5 or 12 days of obstruction, renal macrophage infiltration and tubulointerstitial damage were quantitated. Tissue abundance of Mac-1 and its ligands ICAM-1, RAGE and JAM-C was examined by Western blot and immunoprecipitation. Deficiency of either integrin was associated with reduced early macrophage invasion into the obstructed kidney. After 12 days of UUO, macrophage infiltration and tubulointerstitial injury were reduced only in Mac-1-/- but not in LFA-1-/- mice. Besides ICAM-1, an upregulation of two novel Mac-1 ligands, RAGE and JAM-C were observed, however, with distinct time courses. We conclude that beta2-integrins mediate macrophage infiltration in UUO. Mac-1 is the predominant leukocyte integrin involved in leukocyte recruitment after obstruction. ICAM-1 and its new ligands RAGE and JAM-C are sequentially activated in UUO. Blocking of Mac-1 and its ligands may confer synergistic renoprotective effects in neonatal obstructive nephropathy. PMID- 16374429 TI - Distinct functions for Ras GTPases in the control of proliferation and apoptosis in mouse and human mesangial cells. AB - In previous work, we have demonstrated that Ras GTPases regulate proliferation in a range of human renal cells. The present work compares human and mouse mesangial cell (HMC and MMC) responses to specific knockdown of Ras genes with antisense oligonucleotides (AS-oligos), and examines the role of the p21 (cip1) and p27 (kip1) cyclin-dependent kinase inhibitors in these responses in mouse cells. HMC and MMC were lipofectin transfected with ras-targeted AS-oligo at 200-400 nM for 18 h followed by growth of cells in 20% serum for 18-72 h. Cell proliferation was assessed with an MTS assay and bromodeoxyuridine (BrdU) uptake. Apoptosis was quantified using nuclear stain with Hoechst 33342 dye. In MMC, Ha-ras AS-oligo caused an increase in apoptosis from <2% to 10-15% of cells after 18 h in serum (P<0.01). Control, Ki-ras and N-ras AS-oligos had minimal effects on apoptosis. BrdU uptake studies showed that BrdU+ve MMC were increased by 20-40% (P<0.05) after Ha-ras AS-oligo at 24 h; other ras AS-oligos were inactive. HMC number was reduced by 40-80% (P<0.01) at 48-72 h by both Ha-ras and Ki-ras AS-oligos. These actions were associated with reductions in BrdU+ve cells. In HMC, the ras AS oligos did not induce apoptosis. p21(-,-) MMC showed exaggerated apoptotic responses to Ha-Ras AS-oligo. In mouse cells, Ha-Ras expression appears necessary to prevent apoptotic cell death; Ras expression does not appear necessary for cells to progress through the cell cycle. In human cells, Ras does not appear necessary to prevent apoptosis but Ha-Ras and Ki-Ras appear to be required for cell cycle progression. PMID- 16374430 TI - Renin increases mesangial cell transforming growth factor-beta1 and matrix proteins through receptor-mediated, angiotensin II-independent mechanisms. AB - Recent evidence suggesting a strong interplay between components of the renin angiotensin system and key mediators of fibrosis led us to hypothesize that renin, independent of its enzymatic action to enhance angiotensin (Ang) II synthesis, directly increases production of the fibrogenic cytokine transforming growth factor (TGF)-beta. Human or rat mesangial cells (MCs) were treated with human recombinant renin (HrRenin) or rat recombinant renin (RrRenin) and the effects on TGF-beta1, plasminogen activator inhibitor-type 1 (PAI-1), fibronectin (FN) and collagen 1 mRNA and protein were investigated. Blockade of the rat MC renin receptor was achieved using siRNA. HrRenin or RrRenin, at doses shown to be physiologically relevant, induced marked dose- and time-dependent increases in TGF-beta1. These effects were not altered by adding an inhibitor of renin's enzymatic action (RO 42-5892), the Ang II receptor antagonist losartan or the Ang converting enzyme inhibitor enalapril. RrRenin also induced PAI-1, FN and collagen 1 mRNA and PAI-1 and FN protein in a dose-dependent manner. Neutralizing antibodies to TGF-beta partially blocked these effects. Supernatant and cell lysate Ang I and Ang II levels were extremely low. MC angiotensinogen mRNA was undetectable both with and without added renin. Targeting of the rat renin receptor mRNA with siRNA blocked induction of TGF-beta1. We conclude that renin upregulates MC TGF-beta1 through a receptor-mediated mechanism, independent of Ang II generation or action. Renin-induced increases in TGF-beta1 in turn stimulate increases in PAI-1, FN and collagen I. Thus, renin may contribute to renal fibrotic disease, particularly when therapeutic Ang II blockade elevates plasma renin. PMID- 16374431 TI - Expression of hypoxia-inducible transcription factors in developing human and rat kidneys. AB - Early kidney development is associated with the coordinated branching of the renal tubular and vascular system and hypoxia has been proposed to be a major regulatory factor in this process. Under low oxygen levels, the hypoxia-inducible transcription factor (HIF) regulates the expression of genes involved in angiogenesis, erythropoiesis and glycolysis. To investigate the role of HIF in kidney development, we analyzed the temporal and spatial expression of the oxygen regulated HIF-1alpha and -2alpha subunits at different stages of rat and human kidney development. Using double-staining procedures, localization of the HIF target geneproducts vascular endothelial growth factor (VEGF) and endoglin was studied in relation to HIFalpha. In both species, we found marked nuclear expression of HIF-1alpha in medullary and cortical collecting ducts and in glomerular cells. In contrast, HIF-2alpha was expressed in interstitial and peritubular cells podocytes of the more mature glomeruli. After completion of glomerulogenesis and nephrogenesis, HIF-1alpha and -2alpha were no longer detectable. The HIF-target gene VEGF colocalized with HIF-1alpha protein in glomeruli and medullary collecting ducts. HIF-2alpha colocalized with the endothelium-associated angiogenic factor, endoglin. Both HIFalpha isoforms are activated in the developing kidney in a cell-specific and temporally controlled manner, indicating a regulatory role of oxygen tension in nephrogenesis. HIF 1alpha seems to be primarily involved in tubulogenesis and HIF-2alpha in renal vasculogenesis. Both isoforms are found in glomerulogenesis, potentially having synergistic effects. PMID- 16374432 TI - Heterogeneous mutations in the SLC3A1 and SLC7A9 genes in Chinese patients with cystinuria. AB - Cystinuria is a recessively inherited aminoaciduria that leads to recurrent urolithiasis. It is caused by the defective transport of cystine and dibasic amino acids in the proximal renal tubules and intestinal epithelium. Two genes responsible for this, SLC3A1 and SLC7A9, are known. Patients with two SLC3A1 mutations are classified as type A cystinuria, whereas patients with two SLC7A9 mutations are classified as type B cystinuria. Few clinical and molecular data have been reported for Asian cystinuria patients. In this study, we determined the molecular basis of cystinuria in eight unrelated Chinese subjects. Coding exons and flanking introns of the SLC3A1 and SLC7A9 genes were directly sequenced after amplification by polymerase chain reaction. Five different SLC3A1 mutations were found. Two missense mutations, D210G and S547L, were novel. The other three SLC3A1 mutations (IVS6+2T>C, R181Q and R365W) have been described previously. In addition, four novel SLC7A9 mutations, C137R, c.730delG, IVS10+2_3delTG and IVS12+3insT, together with two previously reported mutations (A70V and G195R) were found. All patients except one carried compound heterozygous mutations. IVS12+3insT was detected in patients from two families. This is the first molecular genetic study on Chinese cystinuria patients. Three patients with type A cystinuria, two with type B cystinuria, and three carriers of type B cystinuria were identified. Our results suggest that the molecular basis of cystinuria is heterogeneous in our local population. PMID- 16374433 TI - A genome-wide linkage scan for genes controlling variation in urinary albumin excretion in type II diabetes. AB - The main hallmark of diabetic nephropathy is elevation in urinary albumin excretion. We performed a genome-wide linkage scan in 63 extended families with multiple members with type II diabetes. Urinary albumin excretion, measured as the albumin-to-creatinine ratio (ACR), was determined in 426 diabetic and 431 nondiabetic relatives who were genotyped for 383 markers. The data were analyzed using variance components linkage analysis. Heritability (h2) of ACR was significant in diabetic (h2=0.23, P=0.0007), and nondiabetic (h2=0.39, P=0.0001) relatives. There was no significant difference in genetic variance of ACR between diabetic and nondiabetic relatives (P=0.16), and the genetic correlation (rG=0.64) for ACR between these two groups was not different from 1 (P=0.12). These results suggested that similar genes contribute to variation in ACR in diabetic and nondiabetic relatives. This hypothesis was supported further by the linkage results. Support for linkage to ACR was suggestive in diabetic relatives and became significant in all relatives for chromosome 22q (logarithm of odds, LOD=3.7) and chromosome 7q (LOD=3.1). When analyses were restricted to 59 Caucasian families, support for linkage in all relatives increased and became significant for 5q (LOD=3.4). In conclusion, genes on chromosomes 22q, 5q and 7q may contribute to variation in urinary albumin excretion in diabetic and nondiabetic individuals. PMID- 16374434 TI - Renal biopsy in congenital ureteropelvic junction obstruction: evidence for parenchymal maldevelopment. AB - The renal histologic changes associated with congenital ureteropelvic junction obstruction (UPJO) and the relationship to clinical imaging have not been well studied. In order to better understand the histologic alterations of congenital UPJO and their relationship with clinical imaging and outcomes, we examined renal biopsies from 61 patients undergoing pyeloplasty for congenital UPJO. Glomeruli were analyzed for various injury patterns and the tubulointerstitium was examined for tubular atrophy/simplification and fibrosis. Two methods were used to evaluate tubular mass: glomerular density and morphometric measurement of tubular size and density. Control specimens were obtained from age-matched autopsy specimens without renal pathology. Glomerular changes were identified in 73% of all biopsies and were present in a range from 1.7 to 91% of glomeruli in each patient. Overt tubulointerstitial changes were present in 26% of all biopsies. Fibrosis was noted to occur with tubulointerstitial changes in a significantly greater fraction of children over the age of 1 year (P=0.026). Increased glomerular density was associated with severe hydronephrosis (P<0.02). Normal glomerular density was inversely correlated with age (P<0.001), but this relationship was more variable in UPJO (P<0.01). Among patients with intact differential function preoperatively (>45%), postoperative functional decline was predicted only by increased glomerular density. 20 biopsies without overt tubulointerstitial changes were analyzed morphometrically and showed a significant reduction in proximal tubular (PT) size, but unchanged density. Distal tubular (DT) size was unchanged in UPJO, but density was increased. The PT/DT ratio was therefore markedly decreased in UPJO (P<0.0001). Both PT and DT sizes were significantly larger in children with a diuretic renogram washout time less than 20 min than those with greater than 20 min, a common threshold for functionally significant obstruction (P<0.05). Capsular thickness was significantly increased in UPJO. In all, 36% of biopsies had a thickness >0.5 mm and this was associated with greater degrees of tubulointerstitial changes and glomerular alterations. Congenital UPJO produces a variety of renal parenchymal changes, which may in part reflect abnormal development. Some of these alternations are seen in clinical imaging and may help predict outcomes, but there is significant discordance between conventional imaging and histological findings. PMID- 16374435 TI - Hemodialysis reduces inhibitory effect of plasma ultrafiltrate on LDL oxidation and subsequent endothelial reactions. AB - Oxidative modification of low-density lipoprotein (LDL) and its deleterious effect on endothelium is implicated in the pathogenesis of atherosclerosis. Endothelium responds to such an insult by upregulating the synthesis of heme oxygenase-1 (HO-1) and ferritin. Endothelial cell damage and dysfunction have been observed in patients with chronic kidney disease (CKD) on maintenance hemodialysis (HD). We studied the effect of low-molecular-weight components of uremic plasma on LDL oxidation and LDL-oxidation-provoked endothelial cell reactions, such as the induction of cytotoxicity and the upregulation of cell protective HO-1 and ferritin. Plasma ultrafiltrate (molecular weight<5000 Da) from CKD patients on HD or when treated conservatively exhibited a pronounced inhibition on heme-mediated oxidative modification of LDL. Endothelial cell cytotoxicity provoked by LDL oxidation was also attenuated by plasma ultrafiltrate from CKD patients. During HD treatment, a dramatic drop occurred in the retardation of oxidative reactions, and a loss of endothelial cytoprotection exerted by plasma ultrafiltrate was noted. The upregulation of HO-1 and ferritin in response to oxidative stress of LDL was blunted by uremic plasma ultrafiltrate that was released by the end of HD. The decreased antioxidant capacity of ultrafiltrate after HD occurred as a consequence of the intradialytic removal of L-ascorbic acid, uric acid, bilirubin, 3-indoxyl sulfate, indoxyl-beta-D glucuronide, p-cresol, and phenol. Intradialytic removal of L-ascorbic acid, uric acid, bilirubin, 3-indoxyl sulfate, indoxyl-beta-D-glucuronide, p-cresol, and phenol increases the risk of LDL oxidation and subsequent endothelial cell damage, which underlines the importance of activation of cytoprotective HO-1 and ferritin in endothelium. PMID- 16374436 TI - Natriuretic peptide and adrenomedullin levels in chronic renal failure and effects of peritoneal dialysis. AB - Plasma levels of B-type natriuretic peptide (BNP) and its N-terminal propeptide (NT-BNP) are elevated in renal impairment and provide a robust prognostic index. The effect of peritoneal dialysis on plasma NT-BNP, however, is unknown. Furthermore, no information exists regarding levels of the N-terminal propeptide for C-type natriuretic peptide (NT-CNP) in renal failure and the effects of peritoneal dialysis. Accordingly, we documented venous levels of these peptides, and adrenomedullin, across peritoneal dialysis. We measured venous BNP, NT-BNP, NT-CNP, adrenomedullin, blood urea nitrogen (BUN) and creatinine before, during and after completion of overnight peritoneal dialysis in 11 patients, and identical sampling was carried out in eight patients (controls) but between peritoneal dialysis treatments. Peptide levels were measured using well validated, published methods. Baseline levels of NT-CNP (212, 150-303 pmol/l, median and 25th and 75th percentiles) were much higher than recorded previously in healthy volunteers or in heart failure, and correlated with plasma creatinine (rs=0.53, P<0.05). Peritoneal dialysis had no effect on plasma NT-CNP, nor on NT BNP, BNP or adrenomedullin (all elevated above normal), whereas both BUN and creatinine levels, as expected, declined (P<0.001). We conclude that plasma levels of NT-CNP are grossly elevated in chronic renal failure and correlated with plasma creatinine, but are not altered by peritoneal dialysis. Likewise, BNP, NT-BNP and adrenomedullin are elevated but are not altered by peritoneal dialysis. This information is needed if levels of these hormones are to be used as prognostic indicators or as a guide to the management of patients with chronic renal failure. PMID- 16374437 TI - A novel approach for accurate prediction of spontaneous passage of ureteral stones: support vector machines. AB - The objective of this study was to optimally predict the spontaneous passage of ureteral stones in patients with renal colic by applying for the first time support vector machines (SVM), an instance of kernel methods, for classification. After reviewing the results found in the literature, we compared the performances obtained with logistic regression (LR) and accurately trained artificial neural networks (ANN) to those obtained with SVM, that is, the standard SVM, and the linear programming SVM (LP-SVM); the latter techniques show an improved performance. Moreover, we rank the prediction factors according to their importance using Fisher scores and the LP-SVM feature weights. A data set of 1163 patients affected by renal colic has been analyzed and restricted to single out a statistically coherent subset of 402 patients. Nine clinical factors are used as inputs for the classification algorithms, to predict one binary output. The algorithms are cross-validated by training and testing on randomly selected train and test-set partitions of the data and reporting the average performance on the test sets. The SVM-based approaches obtained a sensitivity of 84.5% and a specificity of 86.9%. The feature ranking based on LP-SVM gives the highest importance to stone size, stone position and symptom duration before check-up. We propose a statistically correct way of employing LR, ANN and SVM for the prediction of spontaneous passage of ureteral stones in patients with renal colic. SVM outperformed ANN, as well as LR. This study will soon be translated into a practical software toolbox for actual clinical usage. PMID- 16374438 TI - Prevention of uncuffed hemodialysis catheter-related bacteremia using an antibiotic lock technique: a prospective, randomized clinical trial. AB - As a result of the high rate of infection, the NKF-K/DOQI guidelines recommended that an uncuffed catheter (UC) should not be used for longer than three weeks. However, the findings of the Dialysis Outcomes and Practice Patterns Study recognized that 48% of new hemodialysis patients in the US and 75% in Europe used UC for temporary access during arteriovenous fistula or graft maturation. The antibiotic lock technique (ALT) has been recommended to prevent catheter-related bacteremia (CRB). Here, we prospectively evaluated the efficacy of catheter restricted filling using an antibiotic lock solution in preventing CRB. A total of 120 new hemodialysis patients requiring a temporary catheter while waiting for placement and maturation of an arteriovenous fistula or graft were enrolled in this study. Patients with a UC were randomly assigned to receive either an antibiotic-heparin lock solution (antibiotic group: cefazolin 10 mg/ml, gentamicin 5 mg/ml, heparin 1000 U/ml) or a heparin lock solution (no-antibiotic group: heparin 1000 U/ml) as a catheter lock solution during the interdialytic period. The end point of the trial was CRB. CRB developed in seven (11.7%) patients in the no-antibiotic group (Staphylococcus aureus, two; Staphylococcus epidermidis, five) whereas only one patient in the antibiotic group had S. aureus bacteremia. CRB rates per 1000 catheter-days were 0.44 in the antibiotic group versus 3.12 in the no-antibiotic group (P=0.031). Kaplan-Meier analysis also showed that mean CRB-free catheter survival of 59 days (95% CI, 58-61 days) in the antibiotic group was greater than that in the no-antibiotic group (55 days; 95% CI, 50-59 days). The results suggest that ALT may be a beneficial means of reducing the CRB rate in hemodialysis patients with UC. PMID- 16374439 TI - Length polymorphism in heme oxygenase-1 is associated with arteriovenous fistula patency in hemodialysis patients. AB - Heme oxygenase-1 (HO-1) is a rate-limiting enzyme in heme degradation, producing carbon monoxide (CO), which carries potent antiproliferative and anti inflammatory effects in the vascular walls. Transcription of the HO-1 gene is regulated by the length polymorphism of dinucleotide guanosine thymine repeat (GT)(n) in the promoter region, which was measured in this study to determine its association with arteriovenous fistula (AVF) failure in Chinese hemodialysis (HD) patients in Taiwan. L allele means (GT)(n)>or=30 and S allele means (GT)n<30. Therefore, there are two L alleles for L/L genotype, one L and one S allele for L/S genotype, and two S alleles for S/S genotype. Among the 603 HD patients who were enrolled in this study, 178 patients had history of AVF failure, while 425 patients did not. Significant associations were found between AVF failure and the following factors (hazard ratio): longer HD duration (1.004 month), lower pump flow (0.993 ml/min), higher dynamic venous pressure (1.010 mmHg), location of AVF on the right side (1.587 vs left side) and upper arm (2.242 vs forearm), and L/L and L/S genotypes of HO-1 (2.040 vs S/S genotype). The proportion of AVF failure increased from 20.3% in S/S genotype and 31.0% in L/S genotype to 35.4% in L/L genotype (P=0.011). Relative incidences were 1/87.6 (1 episode per 87.6 patient months), 1/129, and 1/224.9 for HD patients with L/L, L/S, and S/S genotypes, respectively (P<0.002). The unassisted patency of AVF at 5 years decreased significantly from 83.8% (124/148) to 75.1% (223/297) and 69% (109/158) in S/S, L/S, and L/L genotypes, respectively (P<0.0001). In comparison with HD patients with S/S genotype, those with L/L genotype had a higher prevalence of coronary artery disease (29.1 vs 14.2%; P=0.005). A longer length polymorphism with (GT)(n) >or=30 in the HO-1 gene was associated with a higher frequency of access failure and a poorer patency of AVF in HD patients. The longer GT repeat in the HO-1 promoter might inhibit gene transcription, and consequently offset the CO mediated protective effect against vascular injury. PMID- 16374440 TI - Renal pathology and retinol status in multiple myeloma patients. AB - Renal dysfunction is a common and serious complication in multiple myeloma (MM) patients. Renal proximal tubule injury is characteristic in MM, and may result in disturbed renal handling of various vitamins. The abnormal excretion of vitamins in urine may result in their low serum levels. The goal of this study was to investigate the urinary excretion of retinol in MM and its relationship with serum retinol concentration. For this purpose, 24 MM patients and 10 healthy individuals were studied. Serum and urinary retinol and retinol-binding protein (RBP) were measured by the high-performance liquid chromatography method and enzyme-linked immunosorbent assay, respectively. The study showed that 58% of MM patients excreted retinol in urine, while only 29% had elevated serum creatinine (P<0.05). There was a strong and highly significant correlation between urinary retinol and RBP (r=0.973, P<0.006). Patients with normal and mildly elevated serum creatinine who excreted retinol in urine had a marked decrease in serum retinol (P<0.007). On the other hand, serum retinol was not decreased in patients with moderate or severe renal failure, despite its urinary loss. Our data indicate that (i) urinary retinol is a more frequent marker of renal dysfunction than elevated serum creatinine in MM patients, (ii) serum retinol is decreased in MM with normal or mildly elevated serum creatinine, but not in patients with moderate/severe renal failure, and (iii) urinary retinol may serve as a diagnostic marker of renal proximal tubule dysfunction in MM patients. PMID- 16374441 TI - Microalbuminuria in patients with previous myocardial infarction. AB - Microalbuminuria (MA) is a cardiovascular risk factor. The aim of this study was to examine the relationship between MA and the intima-media complex of the carotid and brachial artery, endothelial function, glucose metabolism, haemostatic variables and cardiac hypertrophy in patients with coronary heart disease. A total of 123 patients, aged 31-80 years, with a history of previous myocardial infarction and without known diabetes mellitus were examined with B mode ultrasound of common carotid and brachial arteries, flow-mediated dilatation of the brachial artery and echocardiography. A standard oral glucose tolerance test with 75 g of glucose was performed. MA was defined as excretion of 20-200 microg albumin/min. MA was present in 11% of patients. Patients with MA had significantly higher level of 2-h plasma glucose, a lower displacement of the atrioventricular plane, a thicker septum wall and a higher prevalence of impaired glucose tolerance test compared with patients with normoalbuminuria (P<0.05). Urinary albumin excretion (UAE) was significantly and positively associated with calculated intima-media area (cIMa) in both brachial and common carotid arteries as well as with age and interventricular septum thickness. In conclusion, UAE was significantly and positively associated with cIMa in both the common carotid and the brachial arteries as well as with left ventricular septum thickness and glucose intolerance in patients with a history of previous myocardial infarction without known diabetes mellitus. PMID- 16374442 TI - 3-5 year longitudinal follow-up of pediatric patients after acute renal failure. AB - Few data exist regarding the long-term sequelae of acute renal failure (ARF), and these studies are limited to a few renal conditions. We aim to assess the 3-5 year survival and incidence of renal injury in children who previously developed ARF of varying causes. We queried parents, physicians, and hospital/state vital statistics records to find patient survival in 174 children who previously had ARF and survived to hospital discharge. We assessed the following in 29 children for residual renal injury: (a) microalbuminuria, (b) glomerular filtration rate (GFR) by Schwartz formula, (c) hypertension, and (d) hematuria. The 3-5-year survival of children with ARF who survived to hospital discharge was 139/174 (79.9%). Most deaths (24/35 (68.5%)) occurred within 12 months after initial hospitalization. Combining those who died during initial hospitalization and in subsequent 3-5 years, the overall survival rate was 139/245 (56.8%). In all, 16 children progressed to end-stage renal disease; thus, renal survival was 127/173 (91%). Those with primary renal/urologic conditions had lower renal survival than others (24/35 (68.6%) vs 134/139 (96.4%); P<0.0001). Among the 29 patients assessed for long-term sequelae at 3-5 years, 17/29 (59%) subjects had at least one sign of renal injury; microalbuminuria (n=9), hyperfiltration (n=9), decreased GFR (n=4), and hypertension (n=6). A pediatric nephrologist was involved in care of only 6/17 (35%) with chronic renal injury. Patients have high risks of ongoing residual renal injury and death after ARF; therefore, periodic evaluation after the initial insult is necessary. PMID- 16374443 TI - Unilateral multicystic dysplastic kidney. PMID- 16374444 TI - Activated parietal epithelial cells or dedifferentiated podocytes in FSGS: can we make the difference? PMID- 16374445 TI - Use of ultrahigh RAAS blockade: implications for exacerbation of renal failure. PMID- 16374446 TI - Vascular calcification in chronic kidney disease: evolving pathogenesis with progressive chronic kidney disease? PMID- 16374448 TI - The gene for a rare autosomal dominant form of pompholyx maps to chromosome 18q22.1-18q22.3. AB - Pompholyx is a rather common disorder characterized by recurrent crops of vesicles or bullae on the lateral aspects of the fingers, as well as the palms and soles with non-erythematous skin. Until now, very few large families have been reported, so no gene or locus has been identified. Here, we performed a genome-wide search in a large Chinese family to map the chromosome location of the responsible gene. We identified a locus at chromosome 18q22.1-18q22.3 with a maximum two-point LOD score of 3.61 at marker D18S1131 (theta = 0.00). Haplotype analyses indicated that the disease gene is located within 12.07 cM region between markers D18S465 and D18S1362, which corresponds to 8.0 Mb. This is the first locus identified for pompholyx. It will aid future identification of the responsible gene, which will be useful for the understanding of the molecular mechanism of pompholyx. PMID- 16374449 TI - Numerous keratinocyte subtypes involved in wound re-epithelialization. AB - The expression of different keratin intermediate filaments has been used to define keratinocyte maturation and different phenotypic subtypes involved in acute wound (AW) healing. Immunohistochemistry with specific anti-keratin monoclonal and polyclonal antibodies was used to examine AW in normal healthy volunteers (n = 16). In all wounds examined, basal keratinocytes and cells at the leading edge of the wound expressed keratins K5 and K14. However, suprabasal cells had a more complex pattern of keratin expression, which was dependent on their position relative to the wound and location within the suprabasal compartment of the epidermis. In general, K10 was expressed in suprabasal keratinocytes at the wound edge, but not in keratinocytes covering the wound center, which expressed K6, K16, and K17 in a complex fashion. Ki67 expression, a marker of cell proliferation, was restricted to basal and immediate suprabasal layers at the wound edge. Keratinocytes populated the wound bed below the scab by migration, which was supported by keratinocyte proliferation in the surrounding epidermis both at and adjacent to the wound edge. PMID- 16374450 TI - The incidence of both tandem duplications and the common deletion in mtDNA from three distinct categories of sun-exposed human skin and in prolonged culture of fibroblasts. AB - The use of mtDNA damage as a biomarker of cumulative sunlight exposure in human skin is a relatively new field of research. Previous investigations have simply compared the frequency of occurrence of the mtDNA common deletion (CD), and to a much lesser extent that of tandem duplications (TDs), to distinguish between sun protected and sun-exposed skin. This approach is limited because non-melanoma skin cancer is predominantly formed on body sites that are "usually" sun-exposed as opposed to sites that are "occasionally" sun-exposed and as such they differ in their cumulative UV exposure. This study addresses this limitation by investigating the frequency of occurrence of the CD and TDs in 116 age-matched human skin samples taken from three different sun-exposed body sites. There was a greater frequency of the mtDNA damage in "usually" sun-exposed compared to "occasionally" sun-exposed body sites for both the CD and the TDs (P < 0.0001 and P = 0.058, respectively). In addition, we identified a 260 bp triplication of the mtDNA D-loop for the first time in skin. No evidence of the CD or TDs was observed in sun-protected (ie rarely exposed) skin (n = 20). Comparatively little is known about mtDNA damage in prolonged skin cell culture. We have furthered this work by studying the level of the CD and the frequency of the TDs during continued culture of human fibroblasts derived from skin samples taken from usually sun-exposed sites (n = 7 patients). The level of the CD decreases with culture, whereas the frequency of TDs can be maintained. PMID- 16374452 TI - Mitochondrial DNA deletions serve as biomarkers of aging in the skin, but are typically absent in nonmelanoma skin cancers. AB - The potential role of mitochondrial DNA (mtDNA) deletions in nonmelanoma skin cancer (NMSC) and in cutaneous photoaging was explored using a genetic approach. Tumors and photodamaged tumor-free "margin" skin were obtained from NMSC patients undergoing excision and the mtDNA from these specimens was screened for the presence of deletions using long extension PCR. mtDNA deletions were abundant in margin tissue specimens from older patients and their number correlated with the patient age. There was a statistically significant difference between the number of mtDNA deletions in tumors and margins. Fewer deletions were detected in the tumors than the margins and the tumors often had no deletions, implying a potential selection for full-length mtDNA or perhaps a protective role for mtDNA deletions in the process of tumorigenesis. The observed mtDNA deletions from skin were often unreported (19 of 21 deletions), but typically shared structural features with mtDNA deletions reported in other tissues. Some mtDNA deletions were detected from the skin of multiple individuals, including 3,715 and 6,278 base pair (bp) deletions, whose frequencies approached that of the previously well-characterized 4977-bp "common" deletion. These data support the use of mtDNA mutations as biomarkers of photoaging in the skin. PMID- 16374451 TI - Overexpression of laminin-8 in human dermal microvascular endothelial cells promotes angiogenesis-related functions. AB - This study examined the effects of endogenous overexpression of laminin-8 on angiogenesis and wound healing in primary human dermal microvascular endothelial cells (HDMECs). HDMECs expressed laminin-8 and laminin-10, but no other laminins, as determined by radioimmunoprecipitation assay using a panel of antibodies to individual laminin chains. To study laminin-8 function, full-length human laminin alpha4 cDNA was retrovirally transferred to HDMEC, and specific overexpression of laminin-8 was verified by Western blot. Laminin-8 overexpression promoted endothelial cell spreading and migration in scratch assays and accelerated angiogenic tubule formation in collagen gel overlay assays. Strong inhibitory effect of beta1 integrin and weak inhibition by alphavbeta3 integrin antibodies were observed in laminin-8-stimulated cell migration, but only beta1 integrin antibodies affected tubule formation. These studies suggest that laminin-8 overexpression may prove to be a useful method to engineer HDMECs to promote angiogenesis and wound repair. PMID- 16374454 TI - Heterogeneity of T-cell clones infiltrating primary malignant melanomas. AB - It is established that primary malignant melanomas (pMM) can be infiltrated by T cell populations with predominantly one T-cell clone. As pMM generally express multiple tumor-associated antigens (TAA), here we used laser-capture microdissection (LCM) to isolate different tumor-infiltrating lymphocyte (TIL) clusters in order to determine whether pMM are infiltrated only by one single clone or whether the TAA may attract various T-cell populations. As T-cell receptor (TCR) clonality is a useful tool for the demonstration of specific T cell clones, we analyzed 56 pMM, three cutaneous melanoma metastases, and 15 pairs of pMM with a sentinel lymph node (SLN) for clonal rearrangements of the (TCR) gamma chain gene. We detected the clonality of TCR gamma chain gene in 25 of 56 pMM, and in 10 of 17 SLN studied. In four of the 15 pairs of primary tumor and SLN, we found clonal TCR gamma in both the melanoma and the SLN, with two pairs harboring the identical clone. As we detected different clones in pMM and the corresponding SLN, we subsequently performed LCM in 21 malignant melanomas with multiple lymphocytic clusters for the presence of focal clonal T cells in different regions of the melanoma. In seven melanomas, both clusters of TILs showed the same rearranged TCR gamma chain gene and in five of the seven biopsies the clonal rearrangement occurred in different variable (V) regions of the TCR gamma chain gene. These tumors showed infiltration by more than one clone. In 10 biopsies TCR clonality was restricted to one cluster, while the second microdissected sample of the infiltrate was polyclonal. In conclusion, within one primary malignant melanoma several T-cell clones with different rearrangements may occur. The balance between these clones may decide on the progress of melanoma. PMID- 16374453 TI - Epidermal development and wound healing in matrix metalloproteinase 13-deficient mice. AB - Degradation of the extracellular matrix, which is an indispensable step in tissue remodelling processes such as embryonic development and wound healing of the skin, has been attributed to collagenolytic activity of members of the matrix metalloproteinase family (MMPs). Here, we employed mmp13 knockout mice to elucidate the function of MMP13 in embryonic skin development, skin homeostasis, and cutaneous wound healing. Overall epidermal architecture and dermal composition of non-injured skin were indistinguishable from wild-type mice. Despite robust expression of MMP13 in the early phase of wound healing, wild-type and mmp13 knockout animals did not differ in their efficiency of re epithelialization, inflammatory response, granulation tissue formation, angiogenesis, and restoration of basement membrane. Yet, among other MMPs also expressed during wound healing, MMP8 was found to be enhanced in wounds of MMP13 deficient mice. In summary, skin homeostasis and also tissue remodelling processes like embryonic skin development and cutaneous wound healing are independent of MMP13 either owing to MMP13 dispensability or owing to functional substitution by other collagenolytic proteinases such as MMP8. PMID- 16374455 TI - Identification of novel pharmacological activities of an antifungal agent, nystatin, to promote dendritic cell maturation. AB - As an unbiased functional screen to identify agents activating dendritic cells (DCs), we recently developed a DC-based biosensor system, in which a stable murine DC line XS106 was engineered to express the yellow fluorescent protein (YFP) gene under the control of the IL-1beta promoter. Here we report that nystatin (NYT), an antifungal drug of the family of polyene macrolide antibiotics, elevated YFP expression by the resulting XS106-pIL1-YFP DC biosensor clone in a dose-dependent fashion. With respect to the underlying mechanisms, NYT activated the NFkappaB p65 and c-Rel subunits in the parental XS106 DC line. Moreover, NYT dose-dependently increased the surface expression of major histocompatibility complex (MHC) class II (MHC II), CD40, CD54, CD80, and CD86 by murine bone marrow-derived DCs and triggered their robust production of IL-1beta, IL-6, IL-12, tumor necrosis factor alpha, and macrophage inflammatory protein 1alpha. Our results document previously unrecognized pharmacological activities of the most commonly used antifungal drug to promote DC maturation. PMID- 16374456 TI - Germline CDKN2A mutations among Greek patients with early-onset and multiple primary cutaneous melanoma. AB - The genetic basis of melanoma susceptibility among Greek patients is uncharacterized. From 107 consecutive cutaneous melanoma patients, we analyzed the CDKN2A and CDK4 loci among 18 early-onset (< or =40 years) and two multiplex melanoma cases. Overall, we found three CDKN2A mutations (3/20; 15%), including one novel nonsense mutation (Trp110Stop) and two Arg24Pro missense alterations. There were no mutations in ARF or CDK4. CDKN2A mutations are not uncommon among Greek melanoma patients considering that none of the mutation carriers reported a family history of melanoma. PMID- 16374457 TI - Epigenetic inactivation of tumor suppressor genes in serum of patients with cutaneous melanoma. AB - Small amounts of cell-free DNA circulate in both healthy and diseased human blood, while increased concentrations of DNA are present in the serum of cancer patients. Tumor-specific mutations or epigenetic modifications have predominantly been detected in tissue specimens. The purpose of this study was to investigate methylation of five different genes involved in tumor suppression and DNA repair (suppressors of cytokine signaling 1 and 2 (SOCS1, SOCS2)), Ras-association domain family protein 1A (RASSF1a), D-type p16(INK4a) cyclin-dependent kinase inhibitor (CDKN), and O6-methylguanine DNA-methyltransferase (MGMT)) in the serum of 100 patients using methylation-specific PCR. In all, 41 melanoma patients (stage I = 18; stage II = 10; stage III/IV = 13), 13 healthy controls without nevi, and 10 individuals with more than 15 nevi of >5 mm in size were investigated. For comparison, sera from patients with other skin tumors (nine basal cell cancers, five Kaposi's sarcoma), different metastasized cancers (five breast cancers, five colon cancers), and several chronic inflammatory diseases (n = 12) were also analyzed. In addition, we examined if methylation was involved in silencing transcription of these genes in 12 melanoma specimens. SOCS1, SOCS2, RASSF1a, CDKN2a, and MGMT were methylated in 75, 43, 64, 75, and 64% of melanoma samples, respectively. Of the 41 melanoma patients, 83% had one hypermethylated gene, while 66, 51, and 41% had two, three, or four hypermethylated genes, respectively. Also, 20% of these patients showed hypermethylation for all genes, while only 17% showed no methylation. Importantly, the methylation profile of the selected genes from melanoma patients was distinct from the other analyzed tumors. Transcription of SOCS1, SOCS2, CDKN2a, and RASSF1a genes was significantly reduced in fresh melanoma samples, while MGMT showed a 12-fold upregulation at the messenger ribonucleic acid level (P < 0.001). Our findings suggest that epigenetic silencing of the studied tumor suppressor genes is a common and probably important mechanism for melanoma formation. This convenient method using a simple blood sample may contribute to classification of melanoma and awaits clinical validation. PMID- 16374458 TI - Increased neutrophil adherence in psoriasis: role of the human endothelial cell receptor Thy-1 (CD90). AB - The chronic inflammatory skin disease psoriasis is characterized by prominent skin infiltration by neutrophils and microabscess formation. The adhesion of leukocytes and subsequent transmigration through the activated endothelium is one prerequisite for the accumulation of these cells in skin. In recent studies, the human Thy-1 (CD90) was characterized as an adhesion molecule on activated endothelial cells (ECs) mediating the adhesion of neutrophils via the interaction with the beta2-integrin Mac-1. Based on these novel findings, we compared the roles of Thy-1 and ICAM-1 in the adhesion of neutrophils from patients with psoriasis to activated ECs. The adhesion of peripheral blood neutrophils of patients suffering from psoriasis to Thy-1-transfected cells as well as to activated, Thy-1-expressing human dermal microvascular ECs (HDMECs) is distinctly increased in comparison to the adhesion of neutrophils from healthy controls. In contrast, adherence of psoriatic neutrophils to ICAM-1 transfectants is, if at all, only slightly enhanced compared to healthy controls. The interaction of healthy as well as psoriatic polymorphonuclear cells to Thy-1 transfectants and HDMECs was significantly inhibited by blocking Thy-1 on ECs or its receptor Mac-1 on neutrophils, indicating the importance of this interaction for the adhesion of neutrophils to activated endothelium. In conclusion, our data indicate that the adhesion of neutrophils to activated ECs mediated by Thy-1/Mac-1 interaction is an important attachment mechanism facilitating their subsequent migration into lesional psoriatic skin. PMID- 16374459 TI - Regulation of skin microvasculature angiogenesis, cell migration, and permeability by a specific inhibitor of PKCalpha. AB - Activation of protein kinase C (PKC) induces phenotypic changes in the morphology of microvascular endothelial cells that affect major functions of the microvasculature. These functions include the first stages of sprouting in angiogenesis, cell migration following wounding, and vascular permeability. The specific isoform(s) of PKC responsible for each of these changes has not been previously identified. In this study, we used two inflammatory agents, IL-1beta and phorbol myristic acetate, to activate PKC isozymes and specific inhibitors of PKCalpha (Go6976) and PKCbeta (hispidin) to distinguish how each of these isoform(s) controls angiogenesis, wound healing, and permeability. In all cases, only inhibition of PKCalpha inhibited each of these functions when compared to the inhibition of PKCbeta. Additional analysis of the mechanism of action of Go6976 (RT-PCR, Western blots, and immunohistochemistry) of the changes in the phosphorylated and nonphosphorylated forms of PKCalpha in the cell membrane and cytoplasm confirmed the specificity of PKCalpha inhibition by Go6976. These studies therefore indicate a specific and a regulatory role of the PKCalpha isoform in three major endothelial cell functions that are important in the maintenance of microvascular homeostasis. PMID- 16374460 TI - The receptor for advanced glycation end products is highly expressed in the skin and upregulated by advanced glycation end products and tumor necrosis factor alpha. AB - Advanced glycation end products (AGEs) form non-enzymatically from reactions of proteins with reducing sugars. In the skin, AGEs were reported to accumulate in dermal elastin and collagens and to interact nonspecifically with the cell membrane of dermal fibroblasts. Therefore, AGEs may influence the process of skin aging. We investigated the presence of the AGE receptor RAGE in skin and the influence of AGEs on receptor expression and the formation of extracellular matrix (ECM). Sections of sun-protected and sun-exposed skin were analyzed with monoclonal antibodies against (RAGE), heat-shock protein 47, factor XIIIa, CD31, and CD45. RAGE was mainly expressed in fibroblasts, dendrocytes, and keratinocytes and to a minor extent in endothelial and mononuclear cells. Human foreskin fibroblasts (HFFs) highly expressed RAGE on the protein and mRNA level when analyzed by quantitative Western blotting and real-time PCR. Incubation of HFFs with the specific RAGE ligand Nepsilon-(carboxymethyl)lysine-modified BSA (CML-BSA) and tumor necrosis factor-alpha resulted in significant upregulation of RAGE expression. CML-BSA induced a mildly profibrogenic pattern, increasing connective tissue growth factor, transforming growth factor-beta (TGF-beta) 1, and procollagen-alpha1(I) mRNA, whereas expression of matrix metalloproteinase (MMP)-1, -2, -3, and -12 was unaffected. We conclude that in HFFs, AGE-RAGE interactions may influence the process of skin aging through mild stimulation of ECM gene expression. PMID- 16374461 TI - Performance of the SCORTEN during the first five days of hospitalization to predict the prognosis of epidermal necrolysis. AB - The SCORTEN, calculated within 24 hours of admission, is a severity-of-illness score validated for toxic epidermal necrolysis and Stevens-Johnson syndrome. Our purpose was to assess the performance of successive SCORTEN during the first 5 days of hospitalization and to determine the influence of admission delay. Charts of 144 patients aged 46.8 years (+/-19.7), admitted to our department (1993-2003) with Stevens-Johnson syndrome or toxic epidermal necrolysis, were reviewed. Successive SCORTEN were compared between deceased patients (n = 28, 19.4%) and survivors (n = 116). The performance of the score (calibration, discrimination) was assessed on days 1-5. All seven SCORTEN variables, on days 1-5, were associated with a higher mortality rate. The SCORTEN rose slightly during hospitalization, with a significant difference between days 1 and 4 (<0.05). Performance of the SCORTEN was good on each day, but slightly better on day 3. The areas under the receiver-operating characteristic curves were above 80%. The admission delay did not differ between deceased patients and survivors. Delay adjusted SCORTEN was close to the crude SCORTEN. The SCORTEN performance during the first 5 days of hospitalization was excellent, and at its best on day 3. We recommend to compute again the SCORTEN on day 3. The admission delay did not influence prognosis or SCORTEN. PMID- 16374462 TI - A genome scan for epidermal skin pattern in adolescent twins reveals suggestive linkage on 12p13.31. AB - Deterioration in stratum corneum reticular patterning (skin pattern or skin wrinkling) has been associated with increased rates of solar keratoses and skin cancer. A previous analysis of data from the twin sample used in this investigation has shown that 86% of the variation in skin pattern is genetic at age 12 and 62% in an adult sample (mean age = 47.5). Variation due to genetic influences is likely to be influenced by more than one locus. Here, we present results of a genome-wide linkage scan of skin pattern in adolescent twins and siblings from 428 nuclear twin families. Sib-pair linkage analysis was performed on skin pattern data collected from twins at age 12 (378 informative families) and 14 (316 families). Suggestive linkage was found at marker D12S397 (12p13.31, logarithm of the odds (lod) 1.94), when the effect of the trait locus was modelled to influence the skin pattern equally at both ages 12 and 14. In the same analysis, a peak was seen at 4q23 with a lod score of 1.55. A possible candidate for the peak at 12p13.31 is the protease inhibitor, alpha-2 macroglobulin. PMID- 16374463 TI - Chemical peeling by SA-PEG remodels photo-damaged skin: suppressing p53 expression and normalizing keratinocyte differentiation. AB - Chemical peeling with salicylic acid in polyethylene glycol vehicle (SA-PEG), which specifically acts on the stratum corneum, suppresses the development of skin tumors in UVB-irradiated hairless mice. To elucidate the mechanism through which chemical peeling with SA-PEG suppresses skin tumor development, the effects of chemical peeling on photodamaged keratinocytes and cornified envelopes (CEs) were evaluated in vivo. Among UVB-irradiated hairless mice, the structural atypia and expression of p53 protein in keratinocytes induced by UVB irradiation were intensely suppressed in the SA-PEG-treated mice 28 days after the start of weekly SA-PEG treatments when compared to that in the control UVB-irradiated mice. Incomplete expression of filaggrin and loricrin in keratinocytes from the control mice was also improved in keratinocytes from the SA-PEG-treated mice. In photo exposed human facial skin, immature CEs were replaced with mature CEs 4 weeks after treatment with SA-PEG. Restoration of photodamaged stratum corneum by treatment with SA-PEG, which may affect remodeling of the structural environment of the keratinocytes, involved the normalization of keratinocyte differentiation and suppression of skin tumor development. These results suggest that the stratum corneum plays a protective role against carcinogenesis, and provide a novel strategy for the prevention of photo-induced skin tumors. PMID- 16374464 TI - Transcriptional regulation and characterization of the promoter region of the human ABCC6 gene. AB - ABCC6, a member of the adenosine 5'-triphosphate-binding cassette family of genes, encodes multidrug resistance-associated protein 6, a putative transmembrane transporter expressed primarily in the liver and to a significantly lower extent in other tissues. Mutations in ABCC6 result in pseudoxanthoma elasticum, a multi-system heritable connective tissue disorder with variable phenotypic expression. To examine the transcriptional regulation and tissue specific expression of this gene, we cloned 2.6 kb of human ABCC6 promoter and developed a series of 5'-deletion constructs linked to luciferase reporter gene. Transient transfections in a number of cultured cell lines of diverse origin identified a specific NF-kappaB-like sequence (-235/-226), which conferred high level of expression in HepG2 hepatoma cells, inferring liver specificity. The functionality of the promoter fragments was confirmed in vivo by tail vein injection followed by luciferase reporter assay. Testing of selected cytokines revealed that transforming growth factor (TGF)-beta upregulated, while tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma downregulated the promoter activity in HepG2 cells. The responsiveness to TGF-beta was shown to reside primarily within an Sp1/Sp3 cognate-binding site at -58 to -49. The expression of the ABCC6 promoter was also shown to be markedly enhanced by Sp1 protein, as demonstrated by cotransfection of ABCC6 promoter-luciferase constructs and an Sp1 expression vector in Drosophila SL2 cells, which are devoid of endogenous Sp1. Furthermore, four additional transcription factors, with their cognate-binding sequences present in DNA, were shown to bind the 2.6-kb promoter fragment by protein/DNA array. Collectively, the results indicate that human ABCC6 displays tissue-specific gene expression, which can be modulated by proinflammatory cytokines. These findings may have implications for phenotypic expression of heritable and acquired diseases involving abnormality in the ABCC6 gene. PMID- 16374465 TI - The suppressor of cytokine signaling-3 is upregulated in impaired skin repair: implications for keratinocyte proliferation. AB - In this study, we determined regulation and function of the suppressor of cytokine signaling (SOCS)-3 in acute and impaired murine skin repair. Upon skin injury, SOCS-3 was induced and expressed during the inflammatory phase of repair. SOCS-3 protein expression was localized in a subset of non-proliferating keratinocytes within the developing wound margin epithelia. Growth factors (EGF, transforming growth factor-alpha), nitric oxide (NO), and pro-inflammatory cytokines were inducers of SOCS-3 mRNA and protein expression in cultured human (HaCaT) and primary murine keratinocytes. Stable overexpression of SOCS-3 in HaCaT keratinocytes interfered with cytokine-induced signal transducers and activators of transcription-3 phosphorylation and inhibited serum-stimulated proliferation of the cells. Moreover, overexpression of SOCS-3 led to final differentiation of keratinocytes, which was comparable to the Ca(2+)-induced differentiation process in the cells. Finally, we determined SOCS-3 expression in two models of impaired skin repair: NO-deficient and diabetic wound healing. In line with observations from normal repair and SOCS-3 overexpression experiments, reduced keratinocyte proliferation within atrophied neo-epithelia in both models of impaired healing was associated with a marked increase in SOCS-3-expressing wound keratinocytes. In summary, this study suggests a potential novel function of SOCS-3 in regulating keratinocyte proliferation and differentiation in vitro and during skin repair in vivo. PMID- 16374466 TI - Presence of chimeric maternally derived keratinocytes in cutaneous inflammatory diseases of children: the example of pityriasis lichenoides. AB - During pregnancy, maternal cells may enter the fetal circulation and persist until adulthood. The fate of these cells remains unknown. As unexplained T-cell mediated conditions such as pityriasis lichenoides (PL) may occur in children, we aimed at identifying maternal cells in lesional skin of PL and controls. Archived skin biopsy specimens from young males with PL, atopic dermatitis, or normal skin were scanned for the presence of female (presumably maternal) cells using fluorescence in situ hybridization (FISH) with X and Y chromosome-specific probes. Phenotyping of maternal cells relied on FISH combined with anti-CD45, anti-CD1a, or anti-cytokeratin labelling, identifying leukocytes, Langerhans cells, and keratinocytes, respectively. Maternal cells were found in PL (11/12) and controls (4/7), but their average frequency was higher in PL: 99 per million cells as compared to 5 per million cells in controls (P = 0.005). In the epidermis, the maternal microchimeric cells were labelled by anti-cytokeratin in all cases. We identified maternally derived keratinocytes in the skin of male children with inflammatory skin disorders. These cells may either help repair the damaged skin or home initially in the skin and trigger a host (child) versus graft (mother) disease. PMID- 16374467 TI - Alterations in skin and stratified epithelia by constitutively activated PPARalpha. AB - Peroxisome proliferator-activated receptor (PPAR)alpha is a pleiotropic regulator in many cell types and has recently been implicated in skin homeostasis. To determine the role of PPARalpha in skin physiology, transgenic mice were generated using the tetracycline Tet-off regulatory system to target constitutively activated PPARalpha to the epidermis and other stratified epithelia by the bovine keratin K5 promoter. Expression of the transgene during early development resulted in postnatal lethality within 2 days after birth. A thin epidermis, few hair follicles, and abnormal development of the tongue were observed in neonatal transgenic mice. Early mortality was not observed when transgenic PPARalpha expression was diminished by administration of doxycycline (dox) to the mothers. The alterations noted in neonatal mice were not observed in adult mice upon re-expression of the PPARalpha transgene by withdrawing dox. Attenuated hyperplasia of interfollicular epidermis after topical application of the tumor promotor 12-O-tetradecanoyl-phorbol-13-acetate (TPA) was observed in adult mice expressing the PPARalpha transgene. In addition, expression of the PPARalpha transgene in mammary gland during pregnancy resulted in abnormal development of this organ and impaired lactation. Further investigations using primary keratinocytes revealed that expression of the transgene in keratinocytes resulted in increased differentiation and decreased proliferation, which may contribute to the observed phenotype in the transgenic mice. Thus, these results indicate that PPARalpha plays an important role in the development of stratified epithelia including skin, tongue, and mammary gland. PMID- 16374468 TI - Detection of UV-induced pigmentary and epidermal changes over time using in vivo reflectance confocal microscopy. AB - In vivo reflectance confocal microscopy (RCM) provides high-resolution optical sections of the skin in its native state, without needing to fix or section the tissue. Melanin provides an excellent contrast for RCM, giving a bright signal in the confocal images. The pigmented guinea-pig is a common animal model to study human pigment induction and modulation, as its tanning response is comparable to human tanning after exposure to ultraviolet radiation (UVR). We investigated the applicability of RCM to detecting UVR-induced pigmentary changes in this model. Animals were exposed to solar simulator radiation for 7 days. RCM was performed during the irradiation and follow-up period. Compared to non-irradiated skin, an increase in melanocyte size, dendricity, and number, as well as increased pigment in keratinocytes, was seen in the irradiated epidermis. Interestingly, these changes could be detected even before a tanning response was clinically visible. UVR-induced epidermal hyperplasia could also be detected and quantified. In conclusion, in vivo RCM is a sensitive non-invasive imaging technique that can repeatedly measure epidermal pigmentation and thickness, as demonstrated in the guinea-pig model. This technique should greatly enhance our appreciation of dynamic pigmentary changes in human or animal skin over time and in response to specific stimuli. PMID- 16374469 TI - Phenotypic analysis of T-cells in extensive alopecia areata scalp suggests partial tolerance. AB - This study demonstrates the feasibility and efficacy of using flow cytometric analysis with intracellular cytokine staining for characterization of T-cell phenotype and functional status in extensive alopecia areata (EAA) scalp skin. Cell suspensions were made from scalp punch biopsies taken from 12 patients with long-standing EAA (average disease duration 14 years, 95% hair loss) and six control subjects. EAA samples had a lower percentage of CD-3-expressing cells, but CD-4/CD-8 ratios remained similar to controls. Expression of CD-69 was found only in EAA scalp biopsies, suggesting that T-cells from EAA scalp have undergone activation. No difference was found in tumor necrosis factor alpha expression. Surprisingly, EAA scalp T-cells produced less IL-2 and CD-8 T-cells produced less IFN-gamma. Immunohistochemical staining of formalin-fixed paraffin-embedded specimens demonstrated that IFN-gamma-producing cells in EAA scalp were not greater in number than in normal specimens. The few identified IFN-gamma producing cells demonstrated no tendency to localize to the perifollicular region, and were similarly distributed as in control specimens. The abnormalities in cytokine production may explain the relative paucity of inflammatory change observed in the clinical setting and suggest that T-cell responses in EAA scalp are tightly, albeit aberrantly, regulated via mechanisms of peripheral T-cell tolerance. PMID- 16374470 TI - Massive recruitment of type I interferon producing plasmacytoid dendritic cells in varicella skin lesions. PMID- 16374471 TI - Porphyrin formation in actinic keratosis and basal cell carcinoma after topical application of methyl 5-aminolevulinate. AB - Photodynamic therapy using topical methyl 5-aminolevulinate (MAL) is a new treatment modality for basal cell carcinoma (BCC) and actinic keratosis (AK). MAL induces endogenous porphyrins, which act as photosensitizers. Pharmacokinetic studies of the porphyrin-inducing effect of MAL creams (Metvix) applied in different concentrations (16-160 mg/g) and application times are presented. Surface fluorescence measurements were used to monitor porphyrin accumulation in 18 superficial BCCs and 32 AKs. For both lesion types, the fluorescence increased during the first 13 of 28 hours of continuous MAL application. A 20-fold site-to site variation was observed, and there were no significant MAL concentration dependencies. The selectivity between lesions and normal skin was 10-fold during the first hours and decreased throughout the application time. Fluorescence microscopy images of tissue sections from 32 nodular BCCs were analyzed to calculate the porphyrin content in tumor tissue as a function of depth. Significant correlation to MAL concentration was seen within the tumors treated for 3 hours. Increase to 18-hour MAL application enhanced the fluorescence levels in superficial tumor layers, but not in deep layers. In conclusion, application of the 160 mg/g cream for 3 hours gave advantageous porphyrin distributions for all types of lesions. PMID- 16374472 TI - Inflammatory destruction of elastic fibers in acquired cutis laxa is associated with missense alleles in the elastin and fibulin-5 genes. AB - Cutis laxa (CL) is a condition characterized by redundant, pendulous, and inelastic skin. Acquired CL has been reported in patients with inflammatory diseases. The goal of this study was to investigate whether genetic lesions predispose patients to the development of acquired CL. We report a patient who developed CL following a Toxocara canis parasitism. He later had an aortic root aneurysm that required surgical correction. Histological evaluation showed inflammation followed by destruction of elastic fibers in both the skin and the aorta. Mutational analysis showed that the patient was heterozygous for an inherited fibulin-5 (FBLN5) allele G202R and compound heterozygous for elastin (ELN) alleles A55V and G773D. Western blotting indicated abnormal proteolytic processing of tropoelastin (TE) in patient fibroblasts. The FBLN5 202R allele on the other hand led to increased interaction of FBLN5 and TE and increased deposition of insoluble ELN partially rescuing the deficiency conferred by ELN mutation G773D. We demonstrated that the interaction of ELN and FBLN5 alleles results in elastic fibers susceptible to inflammatory destruction. These results suggest that the pathogenesis of acquired CL involves an underlying genetic susceptibility and highlight the importance of molecular genetic analysis in patients with idiopathic connective tissue disorders. PMID- 16374473 TI - Basis for improved permeability barrier homeostasis induced by PPAR and LXR activators: liposensors stimulate lipid synthesis, lamellar body secretion, and post-secretory lipid processing. AB - Previously, we demonstrated that topical applications of peroxisome proliferator activated receptors (PPARs) and liver X receptor (LXR) activators improve permeability barrier homeostasis. We showed further that stimulation of epidermal differentiation provides one mechanism that could account for such improvement. Here, we studied the effects of these agents on the lipid matrix of the stratum corneum. Hairless mice were treated topically with activators of PPARalpha (WY14643), PPARdelta (GW1514), PPARgamma (ciglitazone), and LXR (22(R) cholesterol or TO901317) or vehicle twice daily for 3 days. All activators significantly increased epidermal cholesterol, fatty acid, and sphingolipid synthesis, including the production of barrier-specific ceramide species. In addition, lamellar body (LB) formation, secretion, and post-secretory processing accelerated significantly following acute barrier disruption in PPAR/LXR activator-treated animals. Finally, the activity of epidermal beta glucocerebrosidase, a key lipid-processing enzyme, increased in PPAR/LXR activator-treated animals. Thus, topical PPAR and LXR activators stimulate epidermal lipid synthesis, increase LB secretion, and accelerate extracellular lipid processing, providing additional mechanisms that further account for their ability to improve epidermal permeability barrier homeostasis. Since the liposensors are activated by endogenous lipid metabolites, they may serve as unique regulators of barrier homeostasis. PMID- 16374474 TI - Generation of multiple stable dermcidin-derived antimicrobial peptides in sweat of different body sites. AB - Antimicrobial peptides (AMPs) are effector molecules of innate immunity. Dermcidin (DCD), a recently discovered AMP with broad-spectrum activity, is produced constitutively by the eccrine sweat glands and secreted into sweat. In this study, we investigated the proteolytic processing, site-specific expression, and stability of DCD peptides in eccrine sweat. Using surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) and reversed-phase high-pressure liquid chromatography analysis, we identified in eccrine sweat 14 proteolytically processed DCD peptides. Semiquantitative SELDI TOF-MS analysis indicated that processing of DCD-1L is individually different, but generates a few dominant peptides. At body sites with a high probability for contact with pathogenic microorganisms, a high amount of antimicrobial active DCD peptides was detected in sweat. Furthermore, we show that the secretion rate of DCD is constant during a period of prolonged sweating and that DCD peptides are stable in sweat over several hours. Other known AMPs like the human cathelicidin LL-37 and alpha- or beta-defensins were not detected in significant quantity in eccrine sweat. Owing to the durable and abundant presence, DCD-derived peptides contribute to the first line of defense by building a constant barrier that overlies the epithelial skin. PMID- 16374475 TI - Expression of neutrophil gelatinase-associated lipocalin in skin epidermis. PMID- 16374476 TI - The human dermokine gene: description of novel isoforms with different tissue specific expression and subcellular location. PMID- 16374477 TI - The distal and proximal regulatory regions of the involucrin gene promoter have distinct functions and are required for in vivo involucrin expression. AB - Involucrin is a marker of human keratinocyte differentiation. Previous studies show that the human involucrin gene promoter has two distinct regulatory regions the proximal regulatory region (PRR) and the distal regulatory region (DRR). To study the role of these regions in vivo, we have constructed human involucrin promoter transgenic mice and monitored the impact of specific promoter mutations on involucrin gene expression. In this study, we monitor the impact of specific mutations on expression in a range of surface epithelia. We begin by confirming previous observations made in footpad epidermis by showing that the full-length involucrin promoter drives differentiation-appropriate expression in other surface epithelia, including epidermis, cervix, and esophagus. We further show that mutation of the activator protein AP1-5 site in the DRR completely eliminates transgene expression in all of these tissues. In contrast, mutation of the DRR Sp1 site reduces overall expression, but does not alter the differentiation dependence. Additional studies identify a DRR immediate suprabasal element (ISE). Deletion of the ISE results in a loss of transgene expression in the immediate suprabasal layers. Our studies also indicate that the PRR is important for appropriate transgene expression. Mutation of a PRR C/EBP (CCAAT enhancer binding protein) transcription factor binding site results in patchy/discontinuous expression. These studies suggest that AP1, Sp1, and C/EBP transcription factors are required for appropriate differentiation-dependent involucrin expression, and that the mechanism of regulation is similar in most surface epithelia. PMID- 16374478 TI - SPINK5, the defective gene in netherton syndrome, encodes multiple LEKTI isoforms derived from alternative pre-mRNA processing. AB - The multidomain serine protease inhibitor lymphoepithelial Kazal-type related inhibitor (LEKTI) represents a key regulator of the proteolytic events occurring during epidermal barrier formation and hair development, as attested by the severe autosomal recessive ichthyosiform skin condition Netherton syndrome (NS) caused by mutations in its encoding gene, serine protease inhibitor Kazal-type 5 (SPINK5). Synthesized as a proprotein, LEKTI is rapidly cleaved intracellularly, thus generating a number of potentially bioactive fragments that are secreted. Here, we show that SPINK5 generates three classes of transcripts encoding three different LEKTI isoforms, which differ in their C-terminal portion. In addition to the previously described 15 domain isoform, SPINK5 encodes a shorter LEKTI isoform composed of only the first 13 domains, as well as a longer isoform carrying a 30-amino-acid residue insertion between the 13th and 14th inhibitory domains. We demonstrate that variable amounts of SPINK5 alternative transcripts are detected in all SPINK5 transcriptionally active tissues. Finally, we show that in differentiated cultured human keratinocytes all SPINK5 alternative transcripts are translated into protein and that the LEKTI precursors generate distinct secreted C-terminal proteolytic fragments from a similar cleavage site. Since several data indicate a biological role for the pro-LEKTI-cleaved polypeptides, we hypothesize that the alternative processing of the SPINK5 pre messenger RNA represents an additional mechanism to further increase the structural and functional diversity of the LEKTI bioactive fragments. PMID- 16374479 TI - Investigation of the chromosome 17q25 PSORS2 locus in atopic dermatitis. AB - Psoriasis and atopic dermatitis (AD) are strongly genetic and inherited as multi factorial traits. In both diseases, linkage has been reported to chromosome 17q25. For psoriasis, the locus has been labelled PSORS2. Two peaks of association here contain the psoriasis candidate genes SLC9A3R (solute carrier family 9, isoform 3 regulatory factor), NAT9 (N-acetyltransferase superfamily), and RAPTOR (rapamycin (TOR)). We genotyped 14 of the most significantly associated single-nucleotide polymorphisms (SNPs) in these genes in a panel of 148 families (ECZ1) identified through a proband with active AD. The panel contains 350 siblings and 245 sib-pairs. Replication of positive findings was sought in a second panel, MRC-E, comprising of 278 families, 634 siblings, and 470 sib-pairs. SNP genotyping was carried out by Sequenom MassArray technology. Using family-based tests of association (transmission disequilibrium test), rs878906, in intron 3 of NAT9, was significantly associated with AD (P = 0.010) in the ECZ1 panel. In the MRC-E panel, rs895691, between the end of exon 6 of SLC9A3R1 and exon 7 of NAT9, was associated with AD (P = 0.037). These were not significant when multiple comparisons were taken into account. Haplotype analysis revealed no significant associations in either population. These results suggest that the psoriasis candidate genes do not account for previously observed linkage of the 17q25 PSORS2 locus to AD. PMID- 16374480 TI - Photodynamic therapy does not prevent cutaneous squamous-cell carcinoma in organ transplant recipients: results of a randomized-controlled trial. AB - A randomized-controlled trial with paired observations was performed with 40 organ-transplant recipients to assess the preventive effect of photodynamic therapy (PDT) on the development of new squamous-cell carcinomas and to evaluate the effect of PDT on the number of keratotic skin lesions. The treatment area consisted of a randomly assigned forearm and the corresponding hand, whereas the other forearm and hand served as the control area. After the initial visit, follow-up visits were scheduled at 3-monthly intervals during 2 years. No statistically significant difference was found in the occurrence of new squamous cell carcinomas between the treated and untreated arms: after 2 years of follow up, we observed 15 squamous-cell carcinomas in nine out of 40 PDT-treated arms and 10 squamous-cell carcinomas in nine out of 40 control arms. The number of keratotic skin lesions increased in both arms, but was less pronounced in the PDT treated arm. After 1 year of follow-up, a trend in favor of the PDT-treated arm was observed, but statistical significance was not reached. Nearly 80% of the patients reported mild to severe adverse effects consisting of pain and a burning sensation, immediately after the treatment. No long-term adverse events were noted. In conclusion, PDT does not appear to prevent the occurrence of new squamous-cell carcinomas in organ-transplant recipients, but to some degree, reduces the increase of keratotic skin lesions. PMID- 16374481 TI - Short- and long-wave UV light (UVB and UVA) induce similar mutations in human skin cells. AB - While the mutagenic and carcinogenic properties of longwave UV light (UVA) are well established, mechanisms of UVA mutagenesis remain a matter of debate. To elucidate the mechanisms of mutation formation with UVA in human skin, we determined the spectra of UVA- and UVB-induced mutations in primary human fibroblasts. As with UVB, we found the majority of mutations to be C-to-T transitions also with UVA. For both UVA and UVB, these transitions were found within runs of pyrimidines, at identical hotspots, and with the same predilection for the nontranscribed strand. They also included CC-to-TT tandem mutations. Therefore, these mutations point to a major role of pyrimidine dimers not only in UVB but also in UVA mutagenesis. While some differences were noted, the similarity between the spectra of UVA- and UVB-induced mutations further supports similar mechanisms of mutation formation. A non-dimer type of DNA damage does not appear to play a major role in either UVA or UVB mutagenesis. Therefore, the previously reported increasing mutagenicity per dimer with increasing wavelengths cannot be due to non-dimer DNA damage. Differences in the cellular response to UVA and UVB, such as the less prominent activation of p53 by UVA, might determine a different mutagenic outcome of UVA- and UVB-induced dimers. PMID- 16374482 TI - Upregulation of tenascin-C expression by IL-13 in human dermal fibroblasts via the phosphoinositide 3-kinase/Akt and the protein kinase C signaling pathways. AB - In this study, we examined the genes targeted by IL-13 in human dermal fibroblasts using a cDNA microarray. We focused on the tenascin-C (TN-C) gene, which was identified as one of the genes induced by IL-13. IL-13 induced transcriptional activity of TN-C. IL-13-mediated TN-C expression was inhibited by treatment with wortmannin or LY294002, or Calphostin C. IL-13 induced the phosphorylation of the phosphoinositide 3-kinase (PI3K) regulatory subunit p85, induced tyrosine phosphorylation of Akt, upregulated Akt kinase activity, and activated protein kinase C (PKC)-delta and -epsilon. The IL-13-induced increase in TN-C protein expression was abrogated by the transfection of a dominant negative mutant of Akt, PKC-delta, or PKC-epsilon. In conclusion, we showed that the PI3K/Akt and/or PKC signaling pathways are essential for the IL-13-mediated increase in TN-C. Both serum levels of IL-13 and the expression levels of TN-C in the dermis are increased in patients with systemic sclerosis. Our findings suggest that the expression of TN-C is upregulated in this disease due to IL-13 signaling, and that a blockade of the PI3K or PKC signaling pathway may also have therapeutic value by reducing the amount of TN-C produced during fibrosis. PMID- 16374483 TI - Neurofibromatosis type 1 protein and amyloid precursor protein interact in normal human melanocytes and colocalize with melanosomes. AB - The neurofibromatosis type 1 (NF1) gene product, neurofibromin, is known to interact with Ras, thereby negatively regulating its growth-promoting function. Although this is a well-established interaction, the discovery of other neurofibromin interacting partners could reveal new functional properties of this large protein. Using yeast two-hybrid analysis against a brain cDNA library, we identified a novel interaction between the amyloid precursor protein and the GTPase activating protein-related domain of neurofibromin. This interaction was further analyzed in human melanocytes and confirmed by immunoprecipitation and colocalization studies. In addition, we observed a colocalization of amyloid precursor protein and neurofibromin with melanosomes. Amyloid precursor protein has been proposed to function as a vesicle cargo receptor for the motor protein kinesin-1 in neurons. This colocalization of amyloid precursor protein and neurofibromin with melanosomes was lost in melanocytes obtained from normal skin of a NF1 patient. We suggest that a complex between amyloid precursor protein, neurofibromin, and melanosomes might be important in melanosome transport, which could shed a new light on the etiopathogenesis of pigment-cell-related manifestations in NF1. PMID- 16374504 TI - Tumour necrosis factor-alpha depletes histone deacetylase 1 protein through IKK2. AB - Class I histone deacetylases (HDACs) are ubiquitous enzymes that repress gene expression by deacetylating histone tails and promoting chromatin compaction. Pro inflammatory agents activate programmes of gene expression through transcription factors such as nuclear factor-kappaB (NF-kappaB), even in the context of ubiquitous HDAC activity. How this is accomplished remains unknown. We found that cells treated with the pro-inflammatory cytokine tumour necrosis factor-alpha rapidly and substantially reduced HDAC1 protein levels without affecting other class I HDACs. In addition, HDAC1 depletion occurred through protein degradation, required IKK2 activity and resulted in increased transcription from both NF kappaB-associated and unassociated gene promoters. Our study suggests that the activation of programmes of gene expression by pro-inflammatory agents requires global changes in specific critical epigenetic regulators such as HDAC1. PMID- 16374505 TI - Yeast Trf5p is a nuclear poly(A) polymerase. AB - Recent analyses have shown that the activity of the yeast nuclear exosome is stimulated by the Trf4p-Air1/2p-Mtr4p polyadenylation (TRAMP) complex. Here, we report that strains lacking the Rrp6p component of the nuclear exosome accumulate polyadenylated forms of many different ribosomal RNA precursors (pre-rRNAs). This polyadenylation is reduced in strains lacking either the poly(A) polymerase Trf4p or its close homologue Trf5p. In contrast, polyadenylation is enhanced by overexpression of Trf5p. Polyadenylation is also markedly increased in strains lacking the RNA helicase Mtr4p, indicating that it is required to couple poly(A) polymerase activity to degradation. Tandem affinity purification-tagged purified Trf5p showed polyadenylation activity in vitro, which was abolished by a double point mutation in the predicted catalytic site. Trf5p co-purified with Mtr4p and Air1p, indicating that it forms a complex, designated TRAMP5, that has functions that partially overlap with the TRAMP complex. PMID- 16374506 TI - Redundant pathways for Cdc2 activation in Xenopus oocyte: either cyclin B or Mos synthesis. AB - Xenopus oocytes are arrested in meiotic prophase I. Progesterone induces the resumption of meiotic maturation, which requires continuous protein synthesis to bring about Cdc2 activation. The identification of the newly synthesized proteins has long been a goal. Two plausible candidates have received extensive study. The synthesis of cyclin B and of c-Mos, a kinase that activates the mitogen-activated protein kinase pathway in oocytes, is clearly upregulated by translational control in response to progesterone. Recent studies suggest that ablation of either c-Mos or cyclin B synthesis by antisense oligonucleotides does not block meiotic maturation. Here, however, we show that when both pathways are simultaneously inhibited, progesterone no longer triggers maturation; adding back either c-Mos or cyclin B restores meiotic maturation. We conclude that the specific synthesis of either B-type cyclins or c-Mos, induced by progesterone, is required to induce meiotic maturation. The two pathways seem to be functionally redundant. PMID- 16374507 TI - The involvement of the Mre11/Rad50/Nbs1 complex in the generation of G-overhangs at human telomeres. AB - A central function of telomeres is to prevent chromosome ends from being recognized as DNA double-strand breaks (DSBs). Several proteins involved in processing DSBs associate with telomeres, but the roles of these factors at telomeres are largely unknown. To investigate whether the Mre11/Rad50/Nbs1 (MRN) complex is involved in the generation of proper 3' G-overhangs at human telomere ends, we used RNA interference to decrease expression of MRN and analysed their effects. Reduction of MRN resulted in a transient shortening of G-overhang length in telomerase-positive cells. The terminal nucleotides of both C- and G-rich strands remain unaltered in Mre11-diminished cells, indicating that MRN is not responsible for specifying the final end-processing event. The reduction in overhang length was not seen in telomerase-negative cells, but was observed after the expression of exogenous telomerase, which suggested that the MRN complex might be involved in the recruitment or action of telomerase. PMID- 16374509 TI - Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome. AB - We have determined the human genome to contain 296 different Src homology-3 (SH3) domains and cloned them into a phage-display vector. This provided a powerful and unbiased system for simultaneous assaying of the complete human SH3 proteome for the strongest binding to target proteins of interest, without the limitations posed by short linear peptide ligands or confounding variables of more indirect methods for protein interaction screening. Studies involving three ligand proteins, human immunodeficiency virus-1 Nef, p21-activated kinase (PAK)2 and ADAM15, showed previously reported as well as novel SH3 partners with nanomolar affinities specific for them. This argues that SH3 domains may have a more dominant role in directing cellular protein interactions than has been assumed. Besides showing potentially important new SH3-directed interactions, these studies also led to the discovery of novel signalling proteins, such as the PAK2 binding adaptor protein POSH2 and the ADAM15-binding sorting nexin family member SNX30. PMID- 16374510 TI - Transcriptional regulation of the Neurospora circadian clock gene wc-1 affects the phase of circadian output. AB - WHITE COLLAR-1 (WC-1) is the limiting component of the White Collar Complex (WCC) controlling expression of the Neurospora circadian clock protein Frequency (FRQ). Accumulation of WC-1 is supported by FRQ on a post-transcriptional level. Here, we show that transcription of wc-1 is organized in a complex way. Three promoters drive transcription of wc-1. Pdist is dependent on WCC. Pprox is independent of WCC in darkness, but inducible by light in a WCC-dependent manner. A third promoter, Pint, is located in the wc-1 open reading frame and promotes expression of an amino-terminally truncated WC-1 isoform of unknown function. Expression of wc-1 by Pdist or Pprox alone, or by a heterologous promoter, affects the entrained phase of circadian conidiation and the response of Neurospora to light. Our results indicate that transcriptional regulation of wc-1 is required to modulate the circadian phase of clock output. PMID- 16374511 TI - The Saccharomyces cerevisiae Sae2 protein negatively regulates DNA damage checkpoint signalling. AB - Double-strand breaks (DSBs) elicit a DNA damage response, resulting in checkpoint mediated cell-cycle delay and DNA repair. The Saccharomyces cerevisiae Sae2 protein is known to act together with the MRX complex in meiotic DSB processing, as well as in DNA damage response during the mitotic cell cycle. Here, we report that cells lacking Sae2 fail to turn off both Mec1- and Tel1-dependent checkpoints activated by a single irreparable DSB, and delay Mre11 foci disassembly at DNA breaks, indicating that Sae2 may negatively regulate checkpoint signalling by modulating MRX association at damaged DNA. Consistently, high levels of Sae2 prevent checkpoint activation and impair MRX foci formation in response to unrepaired DSBs. Mec1- and Tel1-dependent Sae2 phosphorylation is necessary for these Sae2 functions, suggesting that the two kinases, once activated, may regulate checkpoint switch off through Sae2-mediated inhibition of MRX signalling. PMID- 16374512 TI - DNA-damage-responsive acetylation of pRb regulates binding to E2F-1. AB - The pRb (retinoblastoma protein) tumour suppressor protein has a crucial role in regulating the G1- to S-phase transition, and its phosphorylation by cyclin dependent kinases is an established and important mechanism in controlling pRb activity. In addition, the targeted acetylation of lysine (K) residues 873/874 in the carboxy-terminal region of pRb located within a cyclin-dependent kinase docking site hinders pRb phosphorylation and thereby retains pRb in an active state of growth suppression. Here, we report that the acetylation of pRb K873/874 occurs in response to DNA damage and that acetylation regulates the interaction between the C-terminal E2F-1-specific domain of pRb and E2F-1. These results define a new role for pRb acetylation in the DNA damage signalling pathway, and suggest that the interaction between pRb and E2F-1 is controlled by DNA-damage dependent acetylation of pRb. PMID- 16374513 TI - A call for the creation of personalized medicine databases. AB - The success of the Human Genome Project raised expectations that the knowledge gained would lead to improved insight into human health and disease, identification of new drug targets and, eventually, a breakthrough in healthcare management. However, the realization of these expectations has been hampered by the lack of essential data on genotype--drug-response phenotype associations. We therefore propose a follow-up to the Human Genome Project: forming global consortia devoted to archiving and analysing group and individual patient data on associations between genotypes and drug-response phenotypes. Here, we discuss the rationale for such personalized medicine databases, and the key practical and ethical issues that need to be addressed in their establishment. PMID- 16374514 TI - The impact of microwave synthesis on drug discovery. AB - In the past few years, using microwave energy to heat and drive chemical reactions has become increasingly popular in the medicinal chemistry community. First described 20 years ago, this non-classical heating method has matured from a laboratory curiosity to an established technique that is heavily used in academia and industry. One of the many advantages of using rapid 'microwave flash heating' for chemical synthesis is the dramatic reduction in reaction times--from days and hours to minutes and seconds. As will be discussed here, there are good reasons why many pharmaceutical companies are incorporating microwave chemistry into their drug discovery efforts. PMID- 16374515 TI - Neuronal eotaxin and the effects of CCR3 antagonist on airway hyperreactivity and M2 receptor dysfunction. AB - Eosinophils cluster around airway nerves in patients with fatal asthma and in antigen-challenged animals. Activated eosinophils release major basic protein, which blocks inhibitory M2 muscarinic receptors (M2Rs) on nerves, increasing acetylcholine release and potentiating vagally mediated bronchoconstriction. We tested whether GW701897B, an antagonist of CCR3 (the receptor for eotaxin as well as a group of eosinophil active chemokines), affected vagal reactivity and M2R function in ovalbumin-challenged guinea pigs. Sensitized animals were treated with the CCR3 antagonist before inhaling ovalbumin. Antigen-challenged animals were hyperresponsive to vagal stimulation, but those that received the CCR3 antagonist were not. M2R function was lost in antigen-challenged animals, but not in those that received the CCR3 antagonist. Although the CCR3 antagonist did not decrease the number of eosinophils in lung tissues as assessed histologically, CCR3 antagonist prevented antigen-induced clustering of eosinophils along the nerves. Immunostaining revealed eotaxin in airway nerves and in cultured airway parasympathetic neurons from both guinea pigs and humans. Both IL-4 and IL-13 increased expression of eotaxin in cultured airway parasympathetic neurons as well as in human neuroblastoma cells. Thus, signaling via CCR3 mediates eosinophil recruitment to airway nerves and may be a prerequisite to blockade of inhibitory M2Rs by eosinophil major basic protein. PMID- 16374516 TI - Macrophage expression of active MMP-9 induces acute plaque disruption in apoE deficient mice. AB - The majority of acute clinical manifestations of atherosclerosis are due to the physical rupture of advanced atherosclerotic plaques. It has been hypothesized that macrophages play a key role in inducing plaque rupture by secreting proteases that destroy the extracellular matrix that provides physical strength to the fibrous cap. Despite reports detailing the expression of multiple proteases by macrophages in rupture-prone regions, there is no direct proof that macrophage-mediated matrix degradation can induce plaque rupture. We aimed to test this hypothesis by retrovirally overexpressing the candidate enzyme MMP-9 in macrophages of advanced atherosclerotic lesions of apoE-/- mice. Despite a greater than 10-fold increase in the expression of MMP-9 by macrophages, there was only a minor increase in the incidence of plaque fissuring. Subsequent analysis revealed that macrophages secrete MMP-9 predominantly as a proform, and this form is unable to degrade the matrix component elastin. Expression of an autoactivating form of MMP-9 in macrophages in vitro greatly enhances elastin degradation and induces significant plaque disruption when overexpressed by macrophages in advanced atherosclerotic lesions of apoE-/- mice in vivo. These data show that enhanced macrophage proteolytic activity can induce acute plaque disruption and highlight MMP-9 as a potential therapeutic target for stabilizing rupture-prone plaques. PMID- 16374517 TI - Inactivation of focal adhesion kinase in cardiomyocytes promotes eccentric cardiac hypertrophy and fibrosis in mice. AB - Focal adhesion kinase (FAK) is a cytoplasmic tyrosine kinase that plays a major role in integrin signaling pathways. Although cardiovascular defects were observed in FAK total KO mice, the embryonic lethality prevented investigation of FAK function in the hearts of adult animals. To circumvent these problems, we created mice in which FAK is selectively inactivated in cardiomyocytes (CFKO mice). We found that CFKO mice develop eccentric cardiac hypertrophy (normal LV wall thickness and increased left chamber dimension) upon stimulation with angiotensin II or pressure overload by transverse aortic constriction as measured by echocardiography. We also found increased heart/body weight ratios, elevated markers of cardiac hypertrophy, multifocal interstitial fibrosis, and increased collagen I and VI expression in CFKO mice compared with control littermates. Spontaneous cardiac chamber dilation and increased expression of hypertrophy markers were found in the older CFKO mice. Analysis of cardiomyocytes isolated from CFKO mice showed increased length but not width. The myocardium of CFKO mice exhibited disorganized myofibrils with increased nonmyofibrillar space filled with swelled mitochondria. Last, decreased tyrosine phosphorylation of FAK substrates p130Cas and paxillin were observed in CFKO mice compared with the control littermates. Together, these results provide strong evidence for a role of FAK in the regulation of heart hypertrophy in vivo. PMID- 16374518 TI - Aberrant maturation of mutant perforin underlies the clinical diversity of hemophagocytic lymphohistiocytosis. AB - Missense mutations in perforin, a critical effector of lymphocyte cytotoxicity, lead to a spectrum of diseases, from familial hemophagocytic lymphohistiocytosis to an increased risk of tumorigenesis. Understanding of the impact of mutations has been limited by an inability to express human perforin in vitro. We have shown, for the first time to our knowledge, that recombinant human perforin is expressed, processed appropriately, and functional in rat basophilic leukemia (RBL) cells following retroviral transduction. Subsequently, we have addressed how perforin missense mutations lead to absent perforin detection and impaired cytotoxicity by analyzing 21 missense mutations by flow cytometry, immunohistochemistry, and immunoblot. We identified perforin missense mutations with partial maturation (class 1), no apparent proteolytic maturation (class 2), and no recognizable forms of perforin (class 3). Class 1 mutations exhibit lytic function when expressed in RBL cells and are associated with residual protein detection and variable cytotoxic function in affected individuals, suggesting that carriers of class 1 alleles may exhibit more subtle immune defects. By contrast, class 3 mutations cause severely diminished perforin detection and cytotoxicity, while class 2 mutations have an intermediate phenotype. Thus, the pathologic mechanism of perforin missense mutation likely involves a protein dosage effect of the mature protein. PMID- 16374519 TI - Suppression of oxidative metabolism and mitochondrial biogenesis by the transcriptional corepressor RIP140 in mouse adipocytes. AB - Using an siRNA-based screen, we identified the transcriptional corepressor RIP140 as a negative regulator of insulin-responsive hexose uptake and oxidative metabolism in 3T3-L1 adipocytes. Affymetrix GeneChip profiling revealed that RIP140 depletion upregulates the expression of clusters of genes in the pathways of glucose uptake, glycolysis, TCA cycle, fatty acid oxidation, mitochondrial biogenesis, and oxidative phosphorylation in these cells. Conversely, we show that reexpression of RIP140 in mouse embryonic fibroblasts derived from RIP140 null mice downregulates expression of many of these same genes. Consistent with these microarray data, RIP140 gene silencing in cultured adipocytes increased both conversion of [14C]glucose to CO2 and mitochondrial oxygen consumption. RIP140-null mice, previously reported to resist weight gain on a high-fat diet, are shown here to display enhanced glucose tolerance and enhanced responsiveness to insulin compared with matched wild-type mice upon high-fat feeding. Mechanistically, RIP140 was found to require the nuclear receptor ERRalpha to regulate hexose uptake and mitochondrial proteins SDHB and CoxVb, although it likely acts through other nuclear receptors as well. We conclude that RIP140 is a major suppressor of adipocyte oxidative metabolism and mitochondrial biogenesis, as well as a negative regulator of whole-body glucose tolerance and energy expenditure in mice. PMID- 16374520 TI - Irs1 and Irs2 signaling is essential for hepatic glucose homeostasis and systemic growth. AB - Insulin receptor substrates, including Irs1 and Irs2, integrate insulin and IGF receptor signals with heterologous pathways to coordinate growth and metabolism. Since Irs2 is thought to be especially important in hepatic nutrient homeostasis, we deleted Irs2 [corrected] from hepatocytes of WT mice (called LKO) or genetically insulin-resistant Irs1-/- mice (called LKO::Irs1-/-). Viable LKO::Irs1-/- mice were 70% smaller than WT or LKO mice, and 40% smaller than Irs1 /- mice. Hepatic insulin receptors were functional in all the mice, but insulin signaling via the Akt-FoxO1 pathway was reduced in Irs1-/- and LKO liver, and undetected in LKO::Irs1-/- liver; however, Gsk3beta phosphorylation (Ser9) and hepatic glycogen stores were nearly normal in all of the mice. LKO and Irs1-/- mice developed insulin resistance and glucose intolerance that never progressed to diabetes, whereas LKO::Irs1-/- mice developed hyperglycemia and hyperinsulinemia immediately after birth. Regardless, few hepatic genes changed expression significantly in Irs1-/- or LKO mice, whereas hundreds of genes changed in LKO::Irs1-/- mice--including elevated levels of Pck1, G6pc, Ppargc1, Pparg, and Igfbp1. Thus, signals delivered by Irs1 or Irs2 regulate hepatic gene expression that coordinates glucose homeostasis and systemic growth. PMID- 16374522 TI - Genome-wide linkage scan to identify Loci for age at first cigarette in Dutch sibling pairs. AB - The heritability of age at first cigarette was estimated in 5883 Dutch twins and siblings registered with the Netherlands Twin Register. Heritability was 60% for males and 39% for females. Shared environmental influences were found in females only (30%). Linkage analyses were performed on data of 422 DZ twins and siblings from 175 families, forming 368 sibling pairs. Genomic regions that may harbor susceptibility loci for age at first cigarette with LOD score greater than 2 were detected on chromosomes 5, 14 and 22. A simultaneous analysis of these three genomic regions showed that most of the variance was explained by the linkage effect on chromosome 5 (205 cM). This peak encloses the D1A dopamine receptor gene which is a functional candidate gene for smoking behavior. PMID- 16374521 TI - ERK1/2 mitogen-activated protein kinase selectively mediates IL-13-induced lung inflammation and remodeling in vivo. AB - IL-13 dysregulation plays a critical role in the pathogenesis of a variety of inflammatory and remodeling diseases. In these settings, STAT6 is believed to be the canonical signaling molecule mediating the tissue effects of IL-13. Signaling cascades involving MAPKs have been linked to inflammation and remodeling. We hypothesized that MAPKs play critical roles in effector responses induced by IL 13 in the lung. We found that Tg IL-13 expression in the lung led to potent activation of ERK1/2 but not JNK1/2 or p38. ERK1/2 activation also occurred in mice with null mutations of STAT6. Systemic administration of the MAPK/ERK kinase 1 (MEK1) inhibitor PD98059 or use of Tg mice in which a dominant-negative MEK1 construct was expressed inhibited IL-13-induced inflammation and alveolar remodeling. There were associated decreases in IL-13-induced chemokines (MIP 1alpha/CCL-3, MIP-1beta/CCL-4, MIP-2/CXCL-1, RANTES/CCL-5), MMP-2, -9, -12, and 14, and cathepsin B and increased levels of alpha1-antitrypsin. IL-13-induced tissue and molecular responses were noted that were equally and differentially dependent on ERK1/2 and STAT6 signaling. Thus, ERK1/2 is activated by IL-13 in the lung in a STAT6-independent manner where it contributes to IL-13-induced inflammation and remodeling and is required for optimal IL-13 stimulation of specific chemokines and proteases as well as the inhibition of specific antiproteases. ERK1/2 regulators may be useful in the treatment of IL-13-induced diseases and disorders. PMID- 16374523 TI - Loss of viability during freeze-thaw of intact and adherent human embryonic stem cells with conventional slow-cooling protocols is predominantly due to apoptosis rather than cellular necrosis. AB - A major challenge in the widespread application of human embryonic stem (hES) cells in clinical therapy and basic scientific research is the development of efficient cryopreservation protocols. Conventional slow-cooling protocols utilizing standard cryoprotectant concentrations i.e. 10% (v/v) DMSO, yield extremely low survival rates of <5% as reported by previous studies. This study characterized cell death within frozen-thawed hES colonies that were cryopreserved under standard conditions. Surprisingly, our results showed that immediately after post-thaw washing, the overwhelming majority of hES cells were viable (approximately 98%), as assessed by the trypan blue exclusion test. However, when the freshly-thawed hES colonies were incubated within a 37 degrees C incubator, there was observed to be a gradual reduction in cell viability over time. The kinetics of cell death was drastically slowed-down by keeping the freshly-thawed hES colonies at 4 degrees C, with >90% of cells remaining viable after 90 min of incubation at 4 degrees C. This effect was reversible upon re exposing the cells to physiological temperature. The vast majority of low temperature-exposed hES colonies gradually underwent cell death upon incubation for a further 90 min at 37 degrees C. Terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end-labeling (TUNEL) assay confirmed apoptosis-induced nuclear DNA fragmentation in frozen-thawed hES cells after incubation at 37 degrees C for 90 min. Expression of active caspase-3 enzyme, which is another prominent marker of apoptosis, was confirmed by immunocytochemical staining, while transmission electron microscopy showed typical ultrastructural features of apoptosis such as chromatin condensation and margination to the nuclear membrane. Hence, our results demonstrated that apoptosis instead of cellular necrosis, is the major mechanism of the loss of viability of cryopreserved hES cells during freeze thawing with conventional slow-cooling protocols. PMID- 16374524 TI - A mouse prostate cancer model induced by Hedgehog overexpression. AB - Hedgehog is a regulatory protein during embryonic development and its abnormal activation in adult tissues has been implicated in tumorigenesis within sites where epithelial-mesenchymal interactions take place. In the prostate, Hedgehog signaling activation was observed during advanced cancer progression and metastasis, but whether Hedgehog overexpression can initiate prostate tumorigenesis remains unknown. We introduced a Hedgehog-expressing vector by intra-prostate injection and electroporation to address the effects of Hedgehog overexpression. The manipulation caused lesions with characteristic prostatic intraepithelial neoplasia or even prostatic cancer (CaP) phenotypes within 30 days, with Hedgehog overexpression demonstrated by immunohistochemistry and Western blot detections. The tumorigenic phenotypes were confirmed by discontinuity of basal cell marker p63, mix-up of CK-8/CK-18 positive epithelial cells in the stoma as well as absence of alpha-SMA positive fibro-muscular sheath. Comparable Hedgehog overexpression was found in human CaP specimen. Thus, Hedgehog overexpression induced prostate tumorigenesis starting from the normal status. Furthermore, a mouse prostate cancer model induced by Hedgehog overexpression was established and may be used for testing novel therapeutical approaches targeting at Hedgehog signaling pathway. PMID- 16374525 TI - Identification of diabetic cardiomyopathy with cardiac magnetic resonance imaging. PMID- 16374526 TI - Imaging of a regressive coronary soft plaque under lipid lowering therapy by multi-slice computed tomography. PMID- 16374527 TI - A novel model to test accuracy and reproducibility of MDCT scan protocols for coronary calcium in vivo. AB - OBJECTIVES: We compared the accuracy and reliability of prospectively triggered, retrospectively ECG gated, and non-gated CT image reconstruction for measurements of coronary artery calcification (CAC) in vivo using a novel animal model. MATERIALS AND METHODS: In six Yorkshire farm pigs, prefabricated chains of cortical bone fragments were sutured over the epicardial bed of the major coronary arteries. Using a 4-slice MDCT scanner, each animal was imaged with two different protocols: sequential acquisition with prospective ECG triggering, and spiral acquisition with retrospectively ECG gated image reconstruction- non-gated reconstructions were also generated from these latter scans. Two independent observers measured the 'Agatston score' (AS), the calcified volume (CV), and mineral mass (MM). To calculate accuracy of MM measurements the ash weight of the burned bone fragments was compared to MDCT derived MM. RESULTS: Six pigs successfully underwent surgery and CT imaging (mean heart rate: 86+/-12 bpm). MM measurements from prospectively ECG triggered CT sequential scans were more accurate (p<0.02) and reproducible (p=0.05) than sequential CT scans without ECG triggering or spiral acquisition using retrospective ECG gating. CONCLUSIONS: At high heart rates prospective ECG triggered image reconstruction is more accurate and reproducible for CAC scoring than retrospective ECG gated reconstruction and non-gated reconstruction. PMID- 16374528 TI - Segmentation in echocardiographic sequences using shape-based snake model combined with generalized Hough transformation. AB - A novel method for segmentation of cardiac structures in temporal echocardiographic sequences based on the snake model is presented. The method is motivated by the observation that the structures of neighboring frames have consistent locations and shapes that aid in segmentation. To cooperate with the constraining information provided by the neighboring frames, we combine the template matching with the conventional snake model. It means that the model not only is driven by conventional internal and external forces, but also combines an additional constraint, the matching degree to measure the similarity between the neighboring prior shape and the derived contour. Furthermore, in order to auto or semi-automatically segment the sequent images without manually drawing the initial contours in each image, generalized Hough transformation (GHT) is used to roughly estimate the initial contour by transforming the neighboring prior shape. The method is particularly useful in case of the large frame-to-frame displacement of structure such as mitral valve. As a result, the active contour can easily detect the desirable boundaries in ultrasound images and has a high penetrability through the interference of various undesirables, such as the speckle, the tissue-related textures and the artifacts. PMID- 16374529 TI - Open access echocardiography - guide or expense factor? PMID- 16374530 TI - Direct access echocardiography. PMID- 16374531 TI - Recent treatment history vs clinical characteristics in the prediction of use of outpatient psychiatric services. AB - BACKGROUND: The use of outpatient psychiatric services has been shown to be a complex function of sociodemographic, clinical, and pathway variables. The relative contribution of each variable or groups of variables in explaining the variability in the use of outpatient psychiatric services, however, remains poorly documented. METHODS: The subjects (N=382) were all patients admitted to an outpatient psychiatric clinic serving mostly a minority and low-income population. The charts of the patients were reviewed for sociodemographic, clinical, and pathway variables and the number of outpatient visits. The pathway variables studied were source of referral and most recent psychiatric treatment service used. Both bivariate and multivariate statistics were used to analyze the data. RESULTS: Pathway variables were better predictors of the number of outpatient visits than clinical variables after controlling for sociodemographic variables. CONCLUSION: Patients recently hospitalized may be sicker or have fewer social supports and therefore require more outpatient visits. Recent treatment history stands out as an important variable in the prediction of the number of outpatient mental health visits. More research is needed to examine the influence of pathway variables on treatment decisions. PMID- 16374532 TI - PEGylated peptide dendrimeric carriers for the delivery of antimalarial drug chloroquine phosphate. AB - PURPOSE: The present study was aimed at developing and exploring the use of uncoated and chondroitin sulfate A (CSA) coated PEGylated poly-L-lysine-based dendrimers for controlled and sustained delivery of a blood schizonticide, chloroquine phosphate (CQ). METHODS: The poly-L-lysine-based peptide dendrimers with PEG amine core prepared and coated with CSA were used to encapsulate the drug molecules by dialysis method. Effect of CSA coating on the surface characteristics, drug entrapment, drug release, stability, hemolytic toxicity, macrophageal interactions, and cytoadherence were determined and compared with those of uncoated systems. RESULTS: The CSA coating of the carriers was found to increase size and drug loading capacity, and reduce drug release rate and hemolytic toxicity. Transmission electron microscopic study revealed the surface properties of the systems. Stability studies had shown increased stability of the formulations on CSA coating. There was a significant reduction in hemolytic toxicity and cytotoxicity of CQ by the present dendrimeric carriers, which became more prominent on further CSA conjugation of the equivalent drug-loaded dendrimeric carriers. There were also significant reduction in levels of ring and trophozoite stages of Plasmodium falciparum in liquid culture when treated with CSA coated dendrimers because of the expression of similar carbohydrate receptors as that by placental and cerebral barriers for infected red blood cells. The systems were also found suitable for prolonging and controlling the blood level of drug as indicated by blood level and organ distribution studies in albino rats on intravenous administration, precluding any significant hematological or toxicological manifestations. CONCLUSION: Thus it can be said that CSA coating can improve drug-loading capacity, control and sustain the release of CQ from such carriers, and can suitably act as safer and effective carriers for intravenous CQ administration. PMID- 16374533 TI - Slower onset of low shear stress leads to less neointimal thickening in experimental vein grafts. AB - Vein grafts respond to low flow and shear stress (tau(w)) by generating thicker walls and smaller lumens through the processes of neointimal hyperplasia and remodeling. Clinically, however, vein grafts with obviously low tau(w), such as those distal to high-grade proximal obstructions, are not infrequently found to be widely patent and pliable. One possible explanation for this phenomenon may be that vein grafts remodel more favorably in response to changes in shear that occur gradually over time compared to abruptly. This hypothesis was tested in an experimental animal model in this report. Two separate models of experimental vein graft failure were created, causing either immediate exposure to ultralow tau(w) (<1 dyne/cm2) or delayed exposure to ultralow tau(w). Under general anesthesia and using a sterile technique, the right external jugular (EJ) veins of 28 New Zealand white rabbits were surgically exposed and isolated. An end-to side distal EJ/common carotid artery anastomosis was created, resulting in a widely patent arteriovenous fistula. For the immediate exposure group (n = 5), the EJ was suture-ligated just proximal to the thoracic inlet, distal to a small 10-50 microm venous tributary. This created a reversed vein segment immediately and abruptly exposed to high wall tension (2.0 +/- 0.3 x 10(4) dyne/cm) and ultralow tau(w) (0.15 +/- 0.08 dyne/cm2). For the delayed exposure group (n = 22), the EJ was ligated over a 0.035 guidewire, leaving a small aperture to sustain some measure of blood flow and tau(w). This predictably resulted in slightly less wall tension (1.4 +/- 0.2 x 10(4) dyne/cm) and higher tau(w) (0.68 +/- 0.21 dyne/cm2) than the immediate exposure group. During the first week, the small outflow aperture in the delayed exposure grafts thrombosed, eventually exposing them to the same low level of tau(w) as the immediate exposure grafts. Thus, the only difference in the two models was that delayed exposure grafts enjoyed a slower decline in tau(w) than immediate exposure grafts. Fourteen rabbits in the delayed exposure group were harvested over the first 7 days to define the patency curve of the restricted outflow channel. As expected, the small aperture had thrombosed in all animals by 7 days. The remaining 14 grafts were harvested after 4 weeks, and 13/14 remained patent. Examination of the hemodynamic parameters at the time of death confirmed that wall tension and tau(w) had equalized (wall tension 0.9 +/- 0.1 vs. 1.1 +/- 0.1 x 10(4) dyne/cm, tau(w) 0.45 +/- 0.12 vs. 0.30 +/- 0.08 dyne/cm2). Histological examination revealed less neointimal hyperplasia in the delayed exposure group compared to the immediate exposure group (wall thickness 266 +/- 16 vs. 180 +/- 24 microm, p = 0.025) as well as a slightly greater luminal diameter (0.30 +/- 0.02 vs. 0.40 +/- 0.02 cm, p = 0.038). The results of this experiment suggest that slow exposure to reduced tau(w) results in more favorable remodeling (less thickening) than abrupt exposure. This finding may explain the occasional clinical observation of a widely patent vein graft even in the face of proximal arterial obstruction and very low flow; the change in tau(w) presumably occurred slowly mitigating the remodeling response. PMID- 16374534 TI - Spontaneous rupture of hypogastric artery aneurysm into the bladder: a case report and review of the literature. AB - Isolated internal iliac artery (hypogastric) aneurysms are rare. The incidence is less than 0.9%. They are usually asymptomatic. Symptoms can be due to compression of adjacent pelvic structures such as the bladder, ureters, and colon. Rupture of hypogastric artery aneurysms into the bladder has rarely been reported and, when present, is associated with prior abdominal instrumentation or trauma. There is one case where an arteriovesicular fistula formed spontaneously without any instrumentation or trauma, similar to our case. We describe an unusual case of an 81-year-old male with spontaneous rupture of a hypogastric artery aneurysm into the urinary bladder presenting as gross hematuria. This case illustrates the importance of a high index of suspicion in diagnosis and early treatment to reduce morbidity and mortality. PMID- 16374535 TI - Examination of the trend in Canada toward geographic centralization of aneurysm surgery during the endovascular era. AB - Unlike in the United States, endovascular aneurysm repair (EVAR) has not been widely disseminated in Canada but has remained limited to large-volume vascular surgery units. Since the development of the endovascular program at our hospital, we have experienced a growth in our aneurysm practice and the area of referral. The purpose of this study was to compare the geographic referral area of our aneurysm practice between 1997 (prior to the introduction of EVAR) and 2003 (EVAR and open surgery). Our prospective database was reviewed to identify patients who underwent elective open aneurysm repair in 1997 and 2003 and those who underwent EVAR in 2003. Each patient's county of residence was identified, allowing for grouping of patients into one of four geographic regions (I-IV) increasingly more distant from our hospital. Proportions were compared with the chi(2) test. In 1997, 105 patients underwent open abdominal aortic aneurysm repair, with the majority of patients originating from the two regions in closest proximity to our hospital (I, 34%; II, 46%; III, 18%; IV, 2%). This contrasts with the 2003 EVAR group (n = 63), which had a higher proportion of patients referred from greater distances (I, 13%; II, 27%; III, 27%; IV, 33%) (p < 0.001). The 2003 open group (n = 165) did not differ statistically with respect to region of origin (I, 18%; II, 41%; III, 21%; IV, 19%) compared to their 2003 EVAR counterparts (p = 0.075) but did have a higher proportion of patients from the more distant regions compared to the 1997 open group (p < 0.001). During the last 5 years, we have experienced a doubling of our elective aneurysm case volumes as well as a trend for patients to be referred from greater distances for both EVAR and open repair. This suggests a trend in Canada toward increased centralization of aneurysm care in centers providing both endovascular and open surgical alternatives. PMID- 16374536 TI - Hemosuccus pancreaticus caused by rupture of a true splenic artery aneurysm following a failure of coil embolization. AB - Hemosuccus pancreaticus, particularly that caused by a primary aneurysm, is rarely encountered. Thus, its clinical characteristics are not well known. We report the case of a 53-year old man, who presented with hemosuccus pancreaticus caused by the rupture of an atherosclerotic aneurysm of the splenic artery and underwent distal pancreatectomy with splenectomy. Only 16 cases of hemosuccus pancreaticus due to primary aneurysm have previously been reported in the English language literature between 1970 and 2003. The relevant literature was also reviewed. The review of the literature showed that because diagnosis is difficult to establish due to intermittent hemorrhage, a long time was often taken before definitive treatment was instituted. Treatment based on a definitive or suspected diagnosis reached a satisfactory result; however, the condition can cause a life threatening situation. When upper gastrointestinal bleeding from an obscure source is encountered, hemosuccus pancreaticus should be considered. We suggest that when an aneurysm of peripancreatic vessels is present, adequate treatment for the aneurysm should be immediately undertaken, even though the site of the bleeding has not been confirmed. PMID- 16374537 TI - TrapEase vena cava filter: a case of filter migration and pulmonary embolism after placement. AB - Inferior vena cava filters provide an alternative method of protection against pulmonary embolism in situations where anticoagulation either fails or is contraindicated. These filters are easily placed, with a relatively minor risk of complications. Currently, we know of only one reported case of filter migration using the TrapEase filter. We present a case report of a migrating TrapEase filter, as well as pulmonary embolism after TrapEase filter placement. This complication developed in a 31-year-old trauma patient who developed bilateral popliteal deep vein thromboses and an initial pulmonary embolus while on low molecular weight heparin. PMID- 16374538 TI - Angiotensin-converting enzyme inhibitor-induced angioedema following carotid endarterectomy misdiagnosed as cervical hematoma. AB - Angioedema is a localized swelling of sudden onset, often associated with angiotensin-converting enzyme (ACE) inhibitor use, that most commonly occurs in the face and neck. We present the case of a patient on long-term ACE inhibitor therapy for hypertension, who developed postoperative angioedema after carotid endarterectomy (CEA). The patient was initially misdiagnosed with an expanding cervical hematoma and underwent a surgical wound reexploration. Angioedema is a rare cause of neck swelling and upper airway distress occurring after CEA in patients treated with ACE inhibitors and should be considered in the differential diagnosis. PMID- 16374539 TI - Mesoaortic compression of the left renal vein (nutcracker syndrome): case reports and review of the literature. AB - Compression of the left renal vein between the aorta and the superior mesenteric artery has been termed the nutcracker syndrome. Obstruction of left renal vein outflow results in venous hypertension with the formation of intra- and extrarenal collaterals and/or the development of gonadal vein reflux. To date, a variety of clinical symptoms due to mesoaortic compression of the left renal vein (nutcracker syndrome) have been described. It is not known what pathophysiological variables play a role in the different clinical manifestations of nutcracker syndrome. We report two patients representing the two different forms of the condition. In the first, hematuria and left flank pain resolved in a young man after successful renocaval reimplantation. In the second, symptoms of pelvic congestion due to pelvic varices improved in a middle-aged woman after successful embolization of the gonadal vein and pelvic collaterals. This report reviews the pathophysiology, presentation, diagnosis including radiographic findings, management options, as well as the current literature on nutcracker syndrome. PMID- 16374540 TI - Induction of endonuclease G-mediated apopotosis in human oral squamous cell carcinoma cells by protein kinase C inhibitor safingol. AB - PKC inhibitor safingol suppressed the growth of human oral squamous cell carcinoma (SCC) cells significantly at concentrations that inhibit PKC isoforms. Safingol inhibited the translocation of PKC following treatment with 12-o tetradecanoylphorbol 13-acetate (TPA) in PKC alpha-EGFP-transfected cells, but not in PKC beta-EGFP- transfected cells, indicating selective inhibition for PKC alpha in oral SCC cells. Flow cytometric analysis and DNA analysis by agarose gel electrophoresis revealed an increase in the proportion of sub-G(1) cells and DNA fragmentation in safingol-treated cells. Mitochondrial membrane potential was decreased, and cytochrome c was released from mitochondria. However, the safingol induced cell death was not accompanied by activation of caspase 3 and cleavage of poly (ADP-ribose) polymerase (PARP). The broad-spectrum caspase inhibitor BD-fmk failed to prevent safingol-induced cell death. Another apoptogenic factor endonuclease G, but not apoptosis-inducing factor (AIF), was also released from mitochondria and translocated to the nucleus. These results suggest that PKC alpha inhibitor safingol induces an endonuclease G- mediated apoptosis in a caspase-independent manner. PMID- 16374541 TI - Fine structural and cytochemical analysis of the processes of cell death of oocytes in atretic follicles in new born and prepubertal rats. AB - The process of cell death of oocytes was studied in atretic ovarian follicles of rats aged from 1 to 28 days using light and electron microscope and cytochemical methods. These methods were TUNEL procedure for DNA breaks, active caspase-3 and lysosome-associated membrane protein 1 (LAMP-1) immunolocalizations. The structural features of the process of oocyte death are mainly characterized by the presence of abundant clear vacuoles and autophagosomes, as well as by the absence of large clumps of compact chromatin associated to the nuclear envelope and apoptotic bodies. These features are common to oocytes in all types of follicles studied. Cytochemical features consisting in positive reactions to TUNEL method, active caspase-3 and LAMP-1 immunolocalizations, are common to the cell death of oocytes in all types of follicles. Particular features of the process of cell death of oocytes are found in different types of follicles. Two morphological patterns of cell death occur in pre-follicular oocytes of the new born and in primordial follicles in 1 to 5 days old rats. One is distinguished by clear nucleoli and moderate compaction of chromatin in clumps frequently resembling meiotic bivalents. The second pattern is characterized by nucleolar condensation and by the absence of compact chromatin. The process of cell death of oocytes in antral follicles is characterized by ribonucleoprotein ribbon-like cytoplasmic structures, pseudo-segmentation, and loss of contact with granulosa cells. PMID- 16374542 TI - Cellular and molecular effects of protons: apoptosis induction and potential implications for cancer therapy. AB - Due to their ballistic precision, apoptosis induction by protons could be a strategy to specifically eliminate neoplastic cells. To characterize the cellular and molecular effects of these hadrons, we performed dose-response and time course experiments by exposing different cell lines (PC3, Ca301D, MCF7) to increasing doses of protons and examining them with FACS, RT-PCR, and electron spin resonance (ESR). Irradiation with a dose of 10 Gy of a 26,7 Mev proton beam altered cell structures such as membranes, caused DNA double strand breaks, and significantly increased intracellular levels of hydroxyl ions, are active oxygen species (ROS). This modified the transcriptome of irradiated cells, activated the mitochondrial (intrinsic) pathway of apoptosis, and resulted in cycle arrest at the G2/M boundary. The number of necrotic cells within the irradiated cell population did not significantly increase with respect to the controls. The effects of irradiation with 20 Gy were qualitatively as well as quantitatively similar, but exposure to 40 Gy caused massive necrosis. Similar experiments with photons demonstrated that they induce apoptosis in a significantly lower number of cells and in a temporally delayed manner. These data advance our knowledge on the cellular and molecular effects of proton irradiation and could be useful for improving current hadrontherapy protocols. PMID- 16374543 TI - Nuclear caspase-3 and caspase-7 activation, and poly(ADP-ribose) polymerase cleavage are early events in camptothecin-induced apoptosis. AB - Chemotherapy-induced apoptosis by DNA-damaging drugs is thought to be generally dependent on the release of cytochrome c and the subsequent activation of caspase 9 and -3. However, the molecular mechanism of how damaged DNA triggers the apoptotic process is not clear. To better understand the mechanisms underlying this process, we examined drug-induced apoptosis in cultured H-460 cells. Using cell fractionation, western blotting, and immunofluorescence assays, we show that the activation of nuclear caspases-7 and -3, and poly(ADP-ribose) polymerase (PARP) cleavage, are early events in camptothecin-induced apoptosis. Moreover, we demonstrate that these events precede the release of cytochrome c and apoptotic inducing factor, and the activation of caspases 2, 8, 9 and 12. Together our results suggest that drugs acting at the DNA level can initiate apoptosis via nuclear caspase activation. PMID- 16374544 TI - Induction of apoptosis in prostate tumor PC-3 cells and inhibition of xenograft prostate tumor growth by the vanilloid capsaicin. AB - Capsaicin, the pungent ingredient of hot chilli pepper, has been recently shown to induce apoptosis in several cell lines through a not well known mechanism. Here, we investigated the role of the vanilloid capsaicin in the death regulation of the human cancer androgen-resistant cell line PC-3. Capsaicin inhibited the growth of PC-3 with an IC(50) of 20 microM cells and induced cell apoptosis, as assessed by flow cytometry and nuclei staining with DAPI. Capsaicin induced apoptosis in prostate cells by a mechanism involving reactive oxygen species generation, dissipation of the mitochondrial inner transmembrane potential (DeltaPsi(m)) and activation of caspase 3. Capsaicin-induced apoptosis was not reduced by the antagonist capsazepine in a dose range from 0.1 microM to 20 microM, suggesting a receptor-independent mechanism. To study the in vivo effects of capsaicinoids, PC-3 cells were grown as xenografts in nude mice. Subcutaneous injection of either capsaicin or capsazepine (5 mg/kg body weight) in nude mice suppressed PC-3 tumor growth in all tumors investigated and induced apoptosis of tumor cells. Our data show a role for capsaicin against androgen-independent prostate cancer cells in vitro and in vivo and suggest that capsaicin is a promising anti-tumor agent in hormone-refractory prostate cancer, which shows resistance to many chemotherapeutic agents. PMID- 16374545 TI - Caspase 8L, a novel inhibitory isoform of caspase 8, is associated with undifferentiated neuroblastoma. AB - Caspase 8 is a key apoptotic factor in the receptor/ligand apoptosis-signaling cascade. Absent caspase 8 expression is shown to correlate with poor prognosis in neuroblastoma. Paradoxically, the caspase 8 gene can produce as plice variant and novel inhibitor of itself-caspase 8l. The presence of caspase 8 alone in tumors may not necessarily portend a good prognosis. We sought to determine whether caspase 8l is present in neuroblastoma and whether over-expression of this protein could inhibit caspase 8-dependent apoptosis. Six of 6 histologically undifferentiated and 2 of 5 differentiated neuroblastoma tumors expressed the caspase 8l isoform, whereas caspase 8l was absent in 3 of 3 ganglioneuromas. Seven human neuroblastoma cell lines were surveyed. Two of the 5 cell lines that expressed caspase 8 also expressed the caspase 8l isoform and both were of a less differentiated neuronal phenotype. Over-expression of caspase 8l in cell lines afforded protection against TRAIL, but not against etoposide induced apoptosis. Conversely, blockade of Caspase 8l in cells that express this splice variant made them more sensitive to apoptosis induced cell death. We demonstrate the caspase 8l isoform is present in neuroblastoma and appears to be associated with undifferentiated cell lines and tumors. Furthermore, it suppresses caspase 8 dependent apoptosis. PMID- 16374546 TI - Short interfering RNA against the PDCD5 attenuates cell apoptosis and caspase-3 activity induced by Bax overexpression. AB - The programmed cell death 5 (PDCD5) protein plays an important apoptosis accelerating role in cells undergoing apoptosis. Decreased expression of PDCD5 has been detected in various human carcinomas. Here we describe that one potent short interfering RNA (siRNA) against the PDCD5 (siPDCD5) specifically inhibits the expression of PDCD5 at both the mRNA and protein level. Cells with decreased PDCD5 expression displayed reduced sensitivity to an apoptotic stimulus induced by Bax overexpression in HeLa, HEK293 and 293T cell lines. Furthermore, we also show that siPDCD5 inhibited both caspase-3 activity and procaspase-3 cleavage. Suppressed expression of PDCD5 attenuates the release of cytochrome c from mitochondria to cytosol induced by Bax overexpression. This phenomenon is accompanied by the reduced translocation of Bax from the cytosol to mitochondria. MTT assay shows that targeted suppression of PDCD5 expression markedly promoted cell proliferation in Hela and HEK293 cell lines. Our data suggests that PDCD5 may exert its effects through pathway of mitochondria by modulating Bax translocation, cytochrome c release and caspase 3 activation directly or indirectly, and that decreased PDCD5 expression may be one of the mechanisms by which tumor cells achieve resistance to apoptotic stimulus induced by anticancer drugs. PMID- 16374547 TI - Mithramycin A activates Fas death pathway in leukemic cell lines. AB - Mithramycin A (MMA, trade name Plicamycin) can facilitate TNFalpha- (Tumor Necrosis Factor) and Fas ligand-induced apoptosis. Besides, several drugs play their anticancer effect through Fas apoptotic pathway. So we investigated the effect of MMA on Fas signaling. In this study we show that MMA induces apoptosis in Fas sensitive Jurkat cells and Fas resistant KG1a cells. This effect involves Fas apoptotic pathway: cell exposure to MMA leads to Fas clustering at the cell surface, DISC (Death Inducing Signaling Complex) formation and caspase cleavage. This phenomenon is independent of Fas ligand/Fas interaction and blockade of Fas death pathway partially inhibits MMA-induced apoptosis. Moreover the activation of Fas apoptotic pathway by MMA is correlated to the modulation of c-Flip(L) expression. Finally, pre-treatment with sub-lethal doses of MMA sensitizes KG1a cells to chemotherapeutic agents. Thus all these results may have important implications to improve clinical treatments. PMID- 16374548 TI - Role of the unfolded protein response in cell death. AB - Unfolded protein response (UPR) is an important genomic response to endoplasmic reticulum (ER) stress. The ER chaperones, GRP78 and Gadd153, play critical roles in cell survival or cell death as part of the UPR, which is regulated by three signaling pathways: PERK/ATF4, IRE1/XBP1 and ATF6. During the UPR, accumulated unfolded protein is either correctly refolded, or unsuccessfully refolded and degraded by the ubiquitin-proteasome pathway. When the unfolded protein exceeds a threshold, damaged cells are committed to cell death, which is mediated by ATF4 and ATF6, as well as activation of the JNK/AP-1/Gadd153-signaling pathway. Gadd153 suppresses activation of Bcl-2 and NF-kappaB. UPR-mediated cell survival or cell death is regulated by the balance of GRP78 and Gadd153 expression, which is coregulated by NF-kappaB in accordance with the magnitude of ER stress. Less susceptibility to cell death upon activation of the UPR may contribute to tumor progression and drug resistance of solid tumors. PMID- 16374549 TI - H2O2 at physiological concentrations modulates Leydig cell function inducing oxidative stress and apoptosis. AB - H(2)O(2) is one of the active reactive oxygen species secreted by macrophages that are seen closely aligned with Leydig cells in the testicular interstitium. The present study was initiated to investigate the role of H(2)O(2) on Leydig cell function in vitro at physiological concentrations. Significant decrease in both testosterone production (p < 0.05) and 3 beta-hydroxysteroid dehydrogenase activity (p < 0.05) in adult Leydig cells were observed even with H(2)O(2) at low concentrations (30 - 50 microM). H(2)O(2) exposure increased oxidative stress in Leydig cells with the rise in lipid peroxidation and fall in the activities of the antioxidant enzymes; superoxide dismutase (SOD), catalase (CAT) & glutathione s-transferase (GST). There was also a marginal increase (approximately 8%) in cell apoptosis accompanied by rise in FasL expression and caspase-3 activation. The above findings indicate that H(2)O(2) as a bio-molecule modulates Leydig cell function at or below physiological concentrations through a variety of actions like decrease in steroidogenic enzyme activity and increase in oxidative stress and apoptosis. PMID- 16374551 TI - Hypoxia-mimetic agents desferrioxamine and cobalt chloride induce leukemic cell apoptosis through different hypoxia-inducible factor-1alpha independent mechanisms. AB - Hypoxia presents pro-apoptotic and anti-apoptotic biphasic effects that appear to be dependent upon cell types and conditions around cells. The substantial reports demonstrated that commonly used hypoxia-mimetic agents cobalt chloride (CoCl(2)) and desferrioxamine (DFO) could also induce apoptosis in many different kinds of cells, but the mechanism was poorly understood. In this work, we compare the apoptosis-inducing effects of these two hypoxia-mimetic agents with acute myeloid leukemic cell lines NB4 and U937 as in vitro models. The results show that both of them induce these leukemic cells to undergo apoptosis with a loss of mitochondrial transmembrane potentials (DeltaPsi m), the activation of caspase 3/8 and the cleavage of anti-apoptotic protein Mcl-1, together with the accumulation of hypoxia-inducible factor-1 alpha (HIF-1alpha) protein, a critical regulator for the cellular response to hypoxia. Metavanadate and sodium nitroprusside significantly abrogate DFO rather than CoCl(2)-induced mitochondrial Delta Psi m collapse, caspase-3/8 activation, Mcl-1 cleavage and apoptosis, but they fail to influence DFO and CoCl(2)-induced HIF-1alpha protein accumulation. Moreover, inducible expression of HIF-1alpha gene dose not alter DFO and CoCl(2)-induced apoptosis in U937 cells. In conclusion, these results propose that although both DFO and CoCl(2)-induced leukemic cell apoptosis by mitochondrial pathway-dependent and HIF-1alpha-independent mechanisms, DFO and CoCl(2)-induced apoptosis involves different initiating signal pathways that remain to be investigated. PMID- 16374552 TI - Cyclin D1 overexpression induces epidermal growth factor-independent resistance to apoptosis linked to BCL-2 in human A431 carcinoma. AB - Overexpression of EGF receptors and constitutive cyclin D1 expression are frequently associated with human squamous carcinomas. We have now investigated whether these parameters influence susceptibility to okadaic acid induced cell death in EGF-receptor overexpressing mutant p53 A431 human carcinoma. Exposure of these cells to 20 nM okadaic acid induced apoptosis-associated caspase 3 activation, DNA fragmentation, cleavage of Poly ADP-Ribose Polymerase (PARP), p53 independent expression of pro-apoptotic bax, and loss of proliferation-promoting cyclin D1. All these alterations were antagonized by concurrent addition of exogenous EGF. Ectopic overexpression of the cyclin D1 gene in A431 carcinoma conferred resistance to 20 nM okadaic acid irrespective of exogenous EGF, associated with a parallel induction of anti-apoptotic bcl-2. Treatment with a subtoxic concentration of a bispecific bcl-2/bcl xL antisense oligonucleotide cooperated with okadaic acid to down-regulate bcl-2 and sensitize cyclin D1 overexpressing cells to okadaic acid. Although EGF protects EGF-R proficient epithelial cells from diverse apoptotic stimuli through Mcl-1, this is the first report demonstrating that cyclin D1 overexpression provides an EGF independent protection from okadaic acid-induced cell death through induction of bcl-2. We also show that this anti-apoptotic effect of cyclin D1 overexpression, can be partly antagonized with antisense strategies that down-regulate anti-apoptotic bcl-2 family members. PMID- 16374550 TI - Protection of human and murine hepatocytes against Fas-induced death by transferrin and iron. AB - Recent studies in a murine model show that transferrin (Tf) interferes with Fas mediated hepatocyte death and liver failure by decreasing pro-apoptotic and increasing anti-apoptotic signals. We show here in vitro in murine and human hepatocyte cell lines and in vivo in mice that Fas-induced apoptosis is modulated by exogenous Tf and iron. The results obtained with iron-free Tf (ApoTf), iron saturated Tf (FeTf), and the iron chelator salicylaldehyde isonicotinoyl hydrazone (SIH) in its iron-free and iron-saturated (FeSIH) forms indicate that apoptosis-modulating effects of Tf are not mediated by iron alone. Both the Tf molecule and iron affect multiple aspects of cell death, and the route of iron delivery to the cell may be critical for the final outcome of cellular Fas signaling. Survival of hepatocytes 'stressed' by Fas signals can be manipulated by Tf and iron and may be a target for prophylactic and therapeutic interventions. PMID- 16374553 TI - Sudden cardiac death unresponsive to implantable defibrillator therapy: an urgent target for clinicians, industry and government. AB - A major expansion in utilization of implantable cardioverter-defibrillators (ICDs) is anticipated based on the results of randomized clinical trials (RCT) that demonstrate increased survival in a sizable population of patients with reduced left ventricular function. However, if RCT accurately reflect clinical practice, then a substantial proportion of patients will die suddenly despite ICD implantation. ICD-unresponsive sudden cardiac death (SCD) has been recognized since the initial ICD experience. Yet, despite 25 years of technical advances, the frequency of ICD-unresponsive SCD has not declined. Pooled analysis of RCT indicates a crude rate of ICD-unresponsive SCD of 5%. This may not cause alarm in an average practice, but it comprises about 30% of cardiac deaths. Meta-analyses of RCT show that ICD therapy is associated with a relative risk reduction of SCD of approximately 60%, far less than the greater than 90% efficacy that many expect. The suboptimal performance of ICD therapy accounts for the failure of some RCT to achieve statistically significant effects on survival. The number of patients with ICD-unresponsive SCD is highly correlated with the number of cardiac deaths among control patients as well as ICD recipients. Otherwise, no definite patterns have emerged that clearly distinguish this mode of demise from other modes of cardiac death. Retrospective post-hoc analyses have not revealed distinguishing characteristics of patients with ICD-unresponsive SCD with respect to clinical variables, pre-terminal symptoms or to the setting of the terminal event. Despite advanced storage capabilities of implanted devices, almost no information has become available from RCT regarding the terminal rhythm or the response of the ICD. These observations have implications for clinical management and research. Candidates for ICD implantation based on RCT should be accurately informed about the residual risk of SCD. Investigators seeking to identify populations likely to benefit from ICD therapy based on SCD incidence should recognize that a significant fraction may not respond to ICD therapy. Reducing the incidence of ICD-unresponsive SCD would substantially improve survival and cost-effectiveness related to ICD therapy. Close cooperation between clinicians, investigators and representatives of industry and government is urgently needed to develop strategies to identify patients prone to ICD-unresponsive SCD, to determine its mechanisms and to develop methods of prevention and treatment. PMID- 16374554 TI - Tpeak-Tend interval as an index of global dispersion of ventricular repolarization: evaluations using monophasic action potential mapping of the epi- and endocardium in swine. AB - The ECG interval from the peak to the end of the T wave (Tpeak-Tend) has been used as an index of transmural dispersion of ventricular repolarization (DVR). The correlation between the Tpeak-Tend interval and the global DVR, however, has not been well-evaluated. METHODS: Monophasic action potentials (MAPs) were recorded from 51+/-10 epicardial and 64 +/- 9 endocardial sites in the left ventricles of 10 pigs, and from 41+/-4 epicardial and 53+/-2 endocardial sites in the right ventricles of 2 of the 10 pigs using the CARTO mapping system. The end of repolarization times over the epi- and endocardium were measured, and the end of repolarization dispersions over the epicardium (DVR-epi), over the endocardium (DVR-endo) and over both (DVR-total) were calculated. The QTpeak, QTend and Tpeak Tend intervals as well as the QTpeak and QTend dispersions were obtained from the simultaneously recorded 12-lead ECG. RESULTS: The maximal Tpeak-Tend intervals (57+/-7 ms) were consistent with the DVR-total (58+/-11 ms, p>0.05), and significantly correlated with the DVR-total (r=0.64, p<0.05). However, the mean Tpeak-Tend intervals (44+/-5 ms), and Tpeak-Tend intervals from lead II (41+/-6 ms) and V5 (43+/-5 ms) were all significantly smaller than and poorly correlated with the DVR-total, as were the QTpeak and QTend dispersions (15+/-2 ms vs. 21+/ 4 ms). CONCLUSION: The maximal Tpeak-Tend interval may be used as a noninvasive estimate for the global DVR, but not the QTpeak and QTend dispersions, nor the mean Tpeak-Tend interval and that from a single lead. PMID- 16374555 TI - The anatomic barriers in the coronary sinus: implications for clinical procedures. AB - BACKGROUND: Coronary sinus (CS) catheterization is often used in cardiac resynchronization therapy. Failure to enter the CS is the most common reason for LV pacing lead implant failure. METHODS: We evaluated the anatomic barriers, Thebesian and Vieussens valves, the CS and its tributaries in 52 adult human cadaver hearts. RESULTS: The average diameter of CS ostiums was 9.47 mm. In 20 of the hearts heavier than 300 g, the average CS os diameter was 10.76 mm, whereas in the remaining hearts was 8.72 mm (p<0.005). The Thebesian valves were observed in 35(67%) of the hearts. In 39(75%) of the hearts Vieussens valves were observed and noted that 6(11%) of them were qualitatively well developed and 33(63%) diminutive. Twenty cases (38%) had 3 vein branches, 19(37%) had 4 branches, 6(11%) had 5 branches, 6(11%) had 2 branches and 1(2%) had 6 branches between great and middle cardiac veins. The anatomic barriers in coronary sinus i.e., Thebesian and Vieussens valves and their branchings were evaluated and found optimal, suboptimal and worst for catheterization in 33, 15 and 4 Thebesian valves; 40, 8, 4 Vieussens valves, respectively. The coronary sinus tributaries between great and middle cardiac veins were found to be optimal, suboptimal and worst for catheterization in 88, 60 and 38 veins, respectively. CONCLUSIONS: Careful evaluation of anatomic barriers is important for treatment success. Thus, knowledge of these functional anatomic features and barriers allows for better utilization of the human coronary sinus for diagnostic and therapeutic purposes. PMID- 16374556 TI - The search for optimal atrioventricular interval. PMID- 16374557 TI - Score indices for predicting electrophysiologic outcomes in patients with unexplained syncope. AB - INTRODUCTION: Our ability to predict a positive electrophysiologic (EP) study in the evaluation of unexplained syncope is suboptimal. AIMS: In patients with unexplained syncope, we defined clinical predictors of bradyarrhythmia and ventricular tachycardia (VT) diagnosed during EP study, constructed diagnostic score indices for bradyarrhythmia and VT, and evaluated the predictive power of each score index. METHODS: All patients evaluated in the Arrhythmia Clinic for unexplained syncope from January 1, 1996, through December 31, 1998, were identified and enrolled in the study. Five hundred eight patients (325 [64%] men; mean+/-SD age, 64+/-17 years) underwent EP testing. We analyzed elements from historical data and noninvasive laboratory findings as predictors of bradyarrhythmia and VT diagnosed on EP study. RESULTS: Fifty-eight patients (11%) had sinus node dysfunction, 94 (19%) had atrioventricular (AV) node disease, 92 (18%) had His-Purkinje system disease, and 101 (20%) had VT. Models were fit using logistic regression analysis. Predictors were assigned weighted scores, and a score index was formulated. The area under the curve associated with sinus node dysfunction, AV node disease, His-Purkinje system disease, and VT models was 0.64, 0.60, 0.84, and 0.60, respectively. CONCLUSIONS: We have constructed diagnostic score indices for EP outcomes of bradyarrhythmia and VT in syncope. Of all the score indices, the model for His-Purkinje system disease has the highest predictive power. PMID- 16374558 TI - 3-dimensional mapping and radiofrequency ablation of atrial flutter in a patient with interrupted inferior vena cava. AB - The presence of isolated interrupted inferior vena cava (IVC) is very rare. Though the occurrence of typical atrial flutter in this setting has recently been described, the use of 3-dimensional activation mapping to aid the management of such patients has not yet been described. We report the successful ablation of this arrhythmia in a 63-year-old woman using the superior route through the internal jugular vein with the help of a mapping system. PMID- 16374559 TI - Cryoablation of a nodoventricular Mahaim fiber. AB - An 11-year old female presented with paroxysmal tachycardia and was diagnosed with a Mahaim fiber during electrophysiologic study. A preexcited tachycardia and the typical variety of AV nodal reentry tachycardia were induced at different times. During preexcited tachycardia, the His bundle electrogram followed the ventricular electrogram, and, introduction of atrial premature beats at different coupling intervals, advanced the peri-AV nodal atrial tissue, with no change in the ventricular cycle length, leading to a diagnosis of an antidromic tachycardia due to a nodoventricular fiber. Cryoablation at a mid-septal location under three dimensional guidance successfully eliminated both tachycardias without detrimental effects to the AV node. PMID- 16374560 TI - Norman J. "Jeff" Holter-"Father" of ambulatory ECG monitoring. PMID- 16374562 TI - Sudden asystole during surgery in the cerebellopontine angle. AB - We report a case of a 40-year-old lady undergoing surgery for a tumor in the cerebellopontine angle. Intraoperatively, patient had a sudden asystole without prior warning sign of bradycardia. It could have been the severe form of trigeminocardiac reflex. The cardiac rhythm returned spontaneously once the surgical manipulation stopped. The remainder of the operation was uneventful and no complication occurred afterwards. The possible mechanism of the event is discussed. PMID- 16374563 TI - Multimodal protocol for awake craniotomy in language cortex tumour surgery. AB - BACKGROUND: Intra-operative neurophysiological language mapping has become an established procedure in patients operated on for tumours in the area of the language cortex. Awake cranial surgery has specific risks and patients are exposed to an increased physical and mental stress. The aim of the study was to establish an algorithm that enables tailoring the neurosurgical and anaesthetic techniques to the individual patient. METHOD: A total of 25 patients underwent awake craniotomy for intra-operative language mapping between 1999 and 2004. Following craniotomy under analgesia and sedation without rigid pin fixation of the head, cortical language mapping was performed in the fully co-operative patient. The results of functional magnetic resonance imaging and of cortical language mapping were incorporated into the 3D dataset for neuronavigation. Depending on the functional data and the individual operative risk tumour resection then proceeded either under conscious sedation with the option of subcortical language monitoring or under general anaesthesia. FINDINGS: After cortical language mapping patients are assigned to one of four groups: BACC (Berlin awake craniotomy criteria) I-IV. BACC I (9 patients): adequate functional data+operative risk not increased-->tumour resection in the awake patient; BACC II (4 patients): limited functional data+operative risk not increased-->tumour resection in the awake patient with the option of language monitoring as needed; BACC III (9 patients): adequate functional data+increased operative risk-->tumour resection under general anaesthesia using functional navigation; BACC IV (3 patients): limited functional data+increased operative risk-->tumour resection in the awake patient with the option of language monitoring as needed. We observed less adverse events in group BACC III. No permanent deterioration of language function occurred in this series. CONCLUSIONS: The multimodal protocol for awake craniotomy provides for tumour resection under general anaesthesia in selected patients using functional neuronavigation. Our experience with the algorithm suggests that it is a useful tool for preserving function in patients undergoing surgery of the language cortex while reducing the operative risk on an individual basis. PMID- 16374564 TI - Entrapped spinal nerve roots in a pseudomeningocoele as a complication of previous spinal surgery. AB - Pseudomeningocele is a rare but well recognised complication of lumbar surgery (microdiscectomy and laminectomy). Most of the patients tolerate the presence of the cyst well, however some present with back pain and spinal claudication, presumably due to neural compression. We report a case who presented following three operations (microdiscectomy, laminectomy and excision of a pseudomeningocele) with symptoms of spinal claudication and bilateral radicular pain. The cause of her pain was evident only at operation and was due to herniation of nerve roots through the dural defect. PMID- 16374565 TI - Role of temozolomide in spinal cord low grade astrocytomas: results in two paediatric patients. AB - BACKGROUND: The optimal treatment of low grade intramedullary spinal cord tumours remains controversial. In many cases the tumours continue to progress even after surgery and radiation. Effective chemotherapy may be an important therapeutic adjunct in this setting. Temozolomide is widely used for brain gliomas, yet its role in the management of spinal cord tumours has not been reported. PROCEDURE: Two paediatric patients with low grade spinal cord astrocytomas were diagnosed to have progression of the tumour in spite of surgery and radiotherapy. They received temozolomide, 200 mg/m2 daily for five days every four weeks for 10 cycles, and were followed serially. RESULTS: Stabilization of the spinal tumour in both patients was observed at 18 months of follow-up. One of the patients developed haematological toxicity requiring platelet transfusion and dose reduction. CONCLUSION: Based on our findings in two paediatric patients, temozolomide may be a useful agent in the management of progressive recurrent low grade spinal cord astrocytomas. PMID- 16374566 TI - The Camino intracranial pressure device in clinical practice. Assessment in a 1000 cases. AB - BACKGROUND: Intracranial pressure (ICP) monitoring has become standard in the management of neurocritical patients. A variety of monitoring techniques and devices are available, each offering advantages and disadvantages. Analysis of large populations has never been performed. PATIENTS AND METHODS: A prospective study was designed to evaluate the Camino fiberoptic intraparenchymal cerebral pressure monitor for complications and accuracy. RESULTS: Between 1992-2004 one thousand consecutive patients had a fiberoptic ICP monitor placed. The most frequent indication for monitoring was severe head injury (697 cases). The average duration of ICP monitoring was 184.6 +/- 94.3 hours; the range was 16-581 hours. Zero drift (range, -17 to 21 mm Hg; mean 7.3 +/- 5.1) was recorded after the devices were removed from 624 patients. Mechanical complications such as: breakage of the optical fiber (n = 17); dislocations of the fixation screw (n = 15) or the probe (n = 13); and failure of ICP recording for unknown reasons (n = 4) were found in 49 Camino devices. CONCLUSIONS: The Camino ICP sensor remains one of the most popular ICP monitoring devices for use in critical neurosurgical patients. The system offers reliable ICP measurements in an acceptable percentage of device complications and the advantage of in vivo recalibration. The incidence of technical complications was low and similar to others devices. PMID- 16374567 TI - Long-term surgical outcome in patients with intracranial hydatid cyst. AB - BACKGROUND: Cerebral hydatid cysts account for up to 3.6% of all intracranial space-occupying lesions, in endemic countries. The vast majority of patients affected are children. Computed tomography (CT) and magnetic resonance imaging (MRI) have greatly contributed to a more accurate diagnosis of hydatids. However, correct pre-operative diagnosis still remains quite puzzling. Extirpation of the intact cyst is the treatment of choice, resulting in most cases to a complete recovery. METHOD: In our retrospective study, we have reviewed 76 cases of intra cranial hydatid disease operated on in our hospital over a 22 year period. Presenting clinical symptoms and signs and the radiological findings on CT and MRI were documented. Albendazole was given preoperatively to patients with giant (>5 cm) or multiple cysts and postoperatively to all patients. The follow-up period ranged from 12 months to 22 years and the outcome was assessed using the Glasgow Outcome Scale (GOS). FINDINGS: Sixty seven (95.7%) of our patients were children. Increased intracranial pressure and papilledema were the predominant findings in this group, whereas focal neurological deficits were most prevalent in adults. CT and MRI revealed round cystic lesions, isodense and iso-intense respectively to cerebrospinal fluid (CSF), with no rim enhancement or perifocal edema. Multiple cysts were identified in 3 cases. Extirpation of the cyst without rupture was accomplished in 56 patients (73.7%). Recurrences occurred in 19 patients (25%). 4 patients (5.3%) died within 6 months after surgery; 3 of these patients had multiple cysts and one died shortly after the operation due to anaphylactic shock following intra-operative rupture of the cyst. CONCLUSION: Long-term follow-up confirms that intracranial hydatid cysts should always be surgically removed without rupture; the outcome remains excellent in these cases. Correct preoperative diagnosis is vital for the successful outcome of surgery. A high index of suspicion is therefore required in endemic areas despite the availability of advanced neuro-imaging. Medical treatment with albendazole seems to be beneficial both pre- and post-operatively. Newer diagnostic methodologies, such as MR spectroscopy and MR diffusion weighted imaging, might lend themselves to the diagnosis of intracranial hydatid cysts. PMID- 16374568 TI - The importance of brainstem mapping in brainstem surgical anatomy before the fourth ventricle and implication for intraoperative neurophysiological mapping. AB - Brain stem mapping (BSM) is an intraoperative neurophysiological procedure to localize cranial motor nuclei on the floor of the fourth ventricle. BSM enables neurosurgeon to understand functional anatomy on the distorted floor of the fourth ventricle, thus, it is emerging as an indispensable tool for challenging brain stem surgery. The authors described the detail of BSM with the special emphasis on its clinical application for the brain stem lesion. Surgical implications based on the result of brains stem mapping would be also informative before planning a brain stem surgery through the floor of fourth ventricle. Despite the recent advancement of MRI to depict the lesion in the brain stem, BSM remains as the only way to provide surgical anatomy in the operative field. BSM could guide a neurosurgeon to the inside of brain stem while preventing direct damage to the cranial motor nuclei on the floor of the fourth ventricle. It is expected that understanding its advantage and limitations would help neurosurgeon to perform safer surgery to the brain stem lesion. PMID- 16374569 TI - Re: The perspective of the International Osteoporosis Foundation on the official positions of the International Society for Clinical Densitometry. PMID- 16374570 TI - Densitometric analysis of the human first tarsometatarsal joint. AB - The first tarsometatarsal (TMT) joint has a complex role in regulating joint pressure in the midfoot. Despite its importance, there have been only a few studies of the functional morphology and biomechanical function of this joint. Here we report about the densitometric pattern of the subchondral bone layer of the articulating elements of the human first TMT joint. We studied dry bones of the first metatarsal and medial cuneiform bones in 64 human cadavers to establish the pattern of the density distribution and to correlate it with the biomechanical function of the joint. The articular surfaces of both the bones were analyzed planimetrically. Half of the specimens (n=32) were sectioned in the sagittal plane and the other 32 articulations were in the transverse plane. In all the sections, the subchondral bone density of the first TMT joint was measured. We found that in sagittal slices the dorsal area of the lateral parts and intermediate parts in females and the dorsal area of the lateral parts, the medial parts and intermediate parts in males were denser than the plantar area and that its density decreased towards the plantar area. The dorsal parts of transverse slices in males were the densest and its density decreased towards the plantar part. In the dorsal, middle and plantar parts in females in transverse slices, the lateral and intermediate areas were denser than the medial areas (P<0.05). There was no significant difference between the bone densities of females and males. PMID- 16374571 TI - Thyroid dimensions of Korean adults on routine neck computed tomography and its relationship to age, sex, and body size. AB - The purpose of this study was to establish a "normal" range of dimensions of the thyroid gland on routine neck computed tomography in the Korean population and to investigate the possible influence of some physiological factors on the thyroid dimension. Neck computed tomography scans of 100 adults (57 males, 43 females; mean age=55.2 years) were reviewed retrospectively to measure the size of the thyroid gland and to evaluate its relationship to the trachea and cervical vertebra. For right and left lobes, the mean width was 15.7+/-2.6 mm and 15.2+/ 3.1 mm, the mean thickness was 20.9+/-3.4 mm and 18.9+/-3.4 mm, the mean length was 61.8+/-8.6 mm and 58.5+/-8.3 mm, the mean estimated volume of each lobe was 8.8+/-3.1 cm(3) and 7.6+/-3.0 cm(3), respectively. The mean volume of total thyroid glands (including isthmus) was 17.5+/-6.6 cm(3). There was no significant difference between the total study group and the normal thyroid function group. The dimensions of the right lobe were significantly larger that those of the left. There was no significant difference between males and females. In multiple regression analysis, it was likely that the influence of body weight on the thyroid dimension was most pronounced. Our study presents initial data for assessing the thyroid gland on neck computed tomography scans in Koreans, and thereby provides suitable limit values of normal thyroid glands. PMID- 16374572 TI - Closer to an ideal solution for inguinal hernia repair: comparison between general surgeons and hernia specialists. AB - BACKGROUND: It is accepted a priori that specialists of hernia surgery have better results than general surgeons who use the same or different techniques as part of their complete surgical repertoire. METHODS: Results of general surgeons trained in the technique of hernia repair using a bilayer connected mesh device (BCMD) was compared to results of specialists and other general surgeons who used other techniques. RESULTS: One report from hernia specialists and three additional reports from trained general surgeons showed similar results using a BCMD. These results were better than results of all other mesh repairs. DISCUSSION: This study shows the value of surgeon-training as well as the value in the design of a particular mesh device for hernia repair. The three components of this device offer three separate, yet connected, barriers to the formation of a recurrent hernia. CONCLUSION: General surgeons trained to use a bilayer mesh device repeatedly duplicated the results of specialists who used it. PMID- 16374573 TI - Bone eburnation in rheumatic diseases: a guiding trace in today's radiological diagnosis and in paleopathology. AB - Bone eburnation is a common anatomical trace of chronic arthropathy. However, its topographical analysis in rheumatic diseases can contribute to knowledge about the latter, by explaining today's diagnosis through radiology as well as by giving an historical perspective through paleopathology. After recalling that eburnated areas can also originate in infectious arthritis, the present analysis consists in a comparison between macroscopic and radiological observations of both osteoarthritis (OA) and rheumatoid arthritis (RA) at an advanced stage. It focuses on the human femoral head because of its demonstrative interest. Two main observations emerge from our study. The eburnated surface is less extensive in OA (where it appears to be essentially linked to the original structure of the hip) and more extensive in RA at an advanced stage (where an additional systemic factor is predominant). The size of the associated osteophytes appears to be inversely proportional to the extent of the corresponding eburnated area. In connection with the OA-RA comparison above, the contribution of the original joint structure to bone eburnation was also illustrated by acromiohumeral eburnation in shoulder OA and by the comparison with dog hip OA. It must also be noted that a femoral head bone remodeling similar on the whole to that of OA can occur in ochronotic arthropathies whose causal chondropathy is due to a genetic defect. Originating in an identified chondropathy, eburnation in ochronotic arthropathy gives us the opportunity to study an OA-type bone remodeling per se as in an experiment supplied by nature and involving a human hip. However, since RA and ochronotic arthropathy are due to a diffuse chondropathy, both may create a similar macroscopic (and thus radiological) eburnation topography. PMID- 16374574 TI - Steroid-reversible parkinsonism as presentation of polymyalgia rheumatica. AB - Polymyalgia rheumatica (PMR) is an inflammatory disorder typically affecting elderly people, characterized by pain and stiffness in the neck and in the shoulder and pelvic girdless with prompt clinical response to low doses of corticosteroids. PMR is closely related to giant cell arteritis (GCA), likely sustained by a "subclinical vasculitis". Whereas in GCA both the central and peripheral nervous systems may be involved, only a PMR case of global, steroid reversible dementia has been hitherto described. We report two elderly patients who abruptly developed, as PMR presenting symptom, an akinetic-rigid parkinsonian syndrome that promptly and completely resolved after corticosteroid treatment. PMID- 16374575 TI - The high prevalence of infections and allergic symptoms in patients with ankylosing spondylitis is associated with clinical symptoms. AB - Ankylosing spondylitis (AS) is strongly associated with the major histocompatibility complex (MHC) class I antigen HLA-B27. This may have influence on the physiologic immune response. Whether it leads to an increased prevalence of infections and/or allergy in AS patients is unclear. This study aims to determine the prevalence of infections and allergic symptoms in patients with AS and to detect a possible association with clinical symptoms. Data on 1,080 AS patients and on 102 disc prolapse patients were collected by questionnaire. The proportion of patients with a symptomatic infection in the last year was 65.5% in AS patients in comparison with 25.5% in disc prolapse patients (p=0.0001). AS patients reported more gastrointestinal (GI) [odds ratio (OR) 5.07, 95% confidence interval (CI) 2.20-11.71], urinary tract (OR 2.81, 95%CI 1.41-5.72), and respiratory (OR 5.83, 95%CI 3.38-10.08) infections than did disc prolapse patients. Multiple infections were more common in AS patients across all infection types. Allergic symptoms were reported by AS patients more frequently than by disc prolapse patients (OR 5.13, 95%CI 3.49-8.80). Patients reporting concurrent inflammatory bowel disease were more likely to report GI (OR 3.0, 95%CI 1.9-4.8) and urinary tract (OR 1.7, 95%CI 1-2.8) infection than primary AS patients. In AS patients, infection was independently associated with female gender (OR 1.96, 95%CI 1.47-2.56), a history of significant peripheral joint inflammation (OR 1.55, 95%CI 1.18-2.05), and increasing pain duration (p=0.05). A high prevalence of common infections and allergic symptoms is seen in patients with AS, most of which are HLA-B27-positive. This may have implications both for underlying mechanisms of disease and for therapeutic options. PMID- 16374576 TI - Efficacy of arthroscopic synovectomy for the effect attenuation cases of infliximab in rheumatoid arthritis. AB - To investigate whether arthroscopic synovectomy is effective for nonresponders to infliximab, anti-tumor necrosis factor-alpha antibody, for the treatment of rheumatoid arthritis (RA), we assessed seven patients including ten arthroscopic synovectomies in knee joint, in shoulder joint, and in ankle joints. We compared C-reactive protein (CRP) and DAS28 (ESR) before and after surgery at 6 and 50 weeks. After arthroscopic synovectomy, we continued the infliximab treatment with methotrexate in a routine manner. We detected synovium proliferation with vascular increase in patellofemoral joint and around the meniscus and femoral and tibial side of the anterior cruciate ligament in the knee joints. We also found synovial proliferation in rotator interval in the glenohumeral joint and fatty changing in subacromial bursa in the shoulder. In the ankle joint, we found synovial proliferation with white meniscoid between tibiofibular joint to develop impingement. Serum CRP was improved from 3.45+/-0.4 to 1.12+/-0.2 at 6 weeks to 1.22+/-0.4 at 50 weeks after arthroscopic synovectomy. There is no severe side effect of arthroscopic synovectomy during infliximab treatment; however, one patient had slight rash that was improved. DAS28 was improved from 5.58+/-0.23 to 3.87+/-0.47 at 6 weeks to 2.58+/-1.49 at 50 weeks after arthroscopic synovectomy. It is possible that arthroscopic synovectomy can be one of the effective methods to continue with the infliximab treatment when its efficacy decreased or in the nonresponders of infliximab for RA patients. PMID- 16374577 TI - Cytoprotective trefoil peptides abound in new functions. PMID- 16374578 TI - Trefoil factors: from ulceration to neoplasia. AB - There is convincing evidence that trefoil factors (TFFs) do play an important role in tumourigenesis. However, their specific roles in cancer are not yet clear. Recently, TFFs have been shown to interfere with crucial biological processes such as cell proliferation, differentiation, apoptosis and angiogenesis. Research on the function of TFFs and its relationship with specific signal transduction pathways has also advanced significantly. As a consequence, some ideas about the role of TFFs in cancer have started to take shape. The objective of this review is to summarize and discuss current knowledge on the relationship between TFFs and cancer. PMID- 16374579 TI - Pleiotropic effects of Trefoil Factor 1 deficiency. AB - Trefoil Factor 1 (TFF1), the first member of the trefoil factor family, is normally expressed in the stomach mucosa. Ectopic expression is also observed in various human pathological conditions, notably in numerous carcinomas and gastrointestinal acute inflammatory disorders. In vivo experimental data using TFF1-deficient mice highlight the pleiotropic functions of TFF1: (i) it is a gastric tumor suppressor gene involved in gastric ontogenesis and homeostasis; (ii) it protects gut mucosa from aggression; (iii) it participates in folding secreted proteins inside the endoplasmic reticulum. At the cellular level, it limits cell proliferation and apoptosis, and favors cell differentiation. Collectively, these data suggest that TFF1 may provide an alternative pharmacological tool for the prevention and treatment of human gastrointestinal diseases. PMID- 16374580 TI - Multiple regulatory pathways for trefoil factor (TFF) genes. AB - The expression of trefoil peptides and their biological consequences are regulated in a multifactorial fashion, and much more work is required in order to fully understand the underlying molecular mechanisms. Central to this will be the identification and functional analysis of trefoil peptide receptors and the full complement of binding proteins. This review summarizes the often fragmentary information available on the environmental, chemical and local regulatory molecules that control trefoil gene expression. Special attention is paid to the nature of the signaling cascades that are activated and the binding proteins that modulate gene transcription. Epigenetic regulation of trefoil gene expression, particularly the role of (de)methylation is described, and the signaling pathways downstream of trefoil peptide activation of target cells are enumerated, as are their physiological and pathological outcomes. PMID- 16374581 TI - Trefoil factors TFF (trefoil factor family) peptide-triggered signals promoting mucosal restitution. AB - Rapid repair of mucous epithelia is essential for preventing inflammation which is a critical component of cancer progression. 'Restitution' is an early repair process which can begin within minutes and is achieved via the migration of neighbouring cells into the wounded area. Mucosal restitution is a multistep process which requires continuous blood flow and includes at least (i) the reduction of cell-cell contacts and a shift in the cell shape towards a migratory phenotype (characteristics of the epithelial-mesenchymal transition), (ii) migration of cells, (iii) repolarization and formation of tight junctions (morphological restitution) and (iv) restoration of barrier function (transmucosal epithelial resistance, functional restitution). Secretory TFF (trefoil factor family) peptides TFF1, TFF2 and TFF3 are well known for their potent protective and healing effects after mucosal damage (function as 'luminal surveillance peptides'). Here, the contributions of the TFFs during the different steps of mucosal restitution are discussed, i. e. the modulation of cell-cell contacts, their motogenic activity and synergy with epidermal growth factor, their anti-apoptotic and pro-angiogenic effects. Special emphasis has been given to discussion of the various signal transduction networks triggered by TFFs. It is becoming increasingly clear that these pathways differ depending on the respective TFF. PMID- 16374582 TI - Trefoil factor family-interacting proteins. AB - Trefoil factor family (TFF) peptides have many in vivo and in vitro effects on restitution, wound healing, apoptosis, cell motility, adhesion and vectorial ion pumping, amongst others. (125)I-TFF peptides bind to cell membranes with classical saturable ability. It would be surprising if there were not TFF-protein interactions that would explain these actions, but to date no convincing TFF binding partner has been shown which unambiguously takes part in any of these functions. Nevertheless, several TFF-binding proteins exist, including the small intestinal CRP-ductin (muclin), which binds TFF2, and the recently described gastric foveolar proteins TFIZ1 (TFF1-binding) and blottin (TFF2-binding), any of which may yet interact in novel ways to elicit TFF-mediated events. This review describes the expression and, where known, the functions of such proteins. PMID- 16374583 TI - Trefoil factor family 2 deficiency and immune response. AB - The protective effect of Trefoil Factor Family (TFF) proteins in the gastrointestinal tract by promoting the healing of injured mucosa is well known. An increasing body of evidence connects TFFs, especially, TFF2 and TFF3, with a possible role in immune regulation. TFF2 is able to inhibit lipopolysaccharide induced nitric oxide production in monocytes and can potently limit leukocyte recruitment at the site of injury. An analysis of gene expression in gastrointestinal tissue of TFF2-deficient mice reveals some new aspects of TFF2's role in the immune response. PMID- 16374584 TI - Structure of mammalian trefoil factors and functional insights. AB - The present review will include the mammalian trefoil factors, TFF1, TFF2 and TFF3. It will summarise the amino acid sequences from different species, their posttranslational modifications and their structures determined by X-ray analysis and nuclear magnetic resonance studies. Trefoil factors all have a well-defined, structurally conserved trefoil domain. The trefoil domain consists of 42 or 43 amino acid residues and contains 6 cysteine residues that form disulphide bonds in a 1-5, 2-4 and 3-6 configuration. By the establishment of an additional intra molecular disulphide bond at the C-terminal end, TFF1 and TFF3 form homodimers or heterodimers. This dimer formation of TFF1 and TFF3 will be discussed, and the possible implications for biological activity will be reviewed. The physicochemical characteristics including protease stability of trefoil factors will be summarised. The biological implications of different molecular forms of trefoil factors and their interaction with mucins will be discussed together with other functional insights. PMID- 16374585 TI - Mechanism of polyamine tolerance in yeast: novel regulators and insights. AB - Polyamines are small charged molecules essential for various cellular functions, but at high levels they are cytotoxic. Two yeast kinases, SKY1 and PTK2, have been demonstrated to regulate polyamine tolerance. Here we report the identification and characterization of additional genes involved in regulating polyamine tolerance: YGL007W, FES1 and AGP2. Deletion of YGL007W, an open reading frame located within the promoter of the membrane proton pump PMA1, decreased Pma1p expression. Deletion of FES1 or AGP2 resulted in reduced polyamine uptake. While high-affinity spermine uptake was practically absent in agp2Delta cells, fes1Delta cells displayed only reduced affinity towards spermine. Despite the reduced uptake, the resistant strains accumulated significant levels of polyamines and displayed increased ornithine decarboxylase activity, suggesting reduced polyamine sensing. Interestingly, fes1Delta cells were highly sensitive to salt ions, suggesting different underlying mechanisms. These results indicate that mechanisms leading to polyamine tolerance are complex, and involve components other than uptake. PMID- 16374586 TI - [Comments on septoplasty]. AB - There is a discrepancy between the occasionally heard opinion that septoplasty is an easy operation and its relatively high failure and complication rates. Here we discuss the specific difficulties and possible pitfalls of functional reconstructive septoplasty. The significance of optimal vision using a binocular operating microscope is emphasized. Most of the important measures needed to avoid postoperative complications such as redeviation, pseudo-hump nose or nasoseptal defect are painstaking subperichondrial dissection without injuring the septal cartilage, use of the so-called Cottle-tunnels, sufficient mobilisation and exact straightening of the anterior septal cartilage without remaining tension, stable reconstruction of the posterior septum with cartilaginous and/or bony fragments after selective resection of deviated septal parts, re-fixation of the mobilized anterior cartilage ("swinging-door") to the periosteum of the anterior nasal spine and, perhaps, primary suture of iatrogenic perforations of the mucoperichondrium during the operation. If postoperative complications such as septal hematoma, septal perforation or saddling of the nose are recognized during the first postoperative week, immediate revision surgery is recommended. PMID- 16374587 TI - Anatomy of the proximal tibiofibular joint. AB - This paper describes the anatomy and function of the proximal tibiofibular joint (PTFJ). The physical dimensions of the joint and the topology of the articular surfaces are described. It is noted that the inclination of the joint is variable, and that joints with a steeper slope away from the transverse plane are less mobile. The ligamentous and tendinous attachments are described. Finally, the histological features of the articular surfaces are presented. The clinical importance of the anatomical features is discussed. PMID- 16374588 TI - Minimally invasive endoscopic reconstruction technique of acute AC-joint dislocations: a cadaver study. AB - The treatment of acute Rockwood type III AC-joint dislocations is controversial. Problems related to open surgery are soft tissue healing, residual instability and the necessity of hardware removal. After non-operative therapy the cosmetic result may be problematic and in some cases symptomatic instabilities occur. The goal of the present cadaver study was to develop a new, minimally invasive technique for acute AC-joint reconstructions and to analyse its potential risk for neurovascular injuries. The surgical technique was based on an arthroscopically assisted reconstruction of the coracoclavicular ligaments with a suture anchor (Arthrex, Naples, FL, USA) and a supplemental stabilization of the AC-joint capsule with a suture cerclage (Fibre Wire 2, Arthrex) performed on ten cadaveric shoulder specimens. After surgery all specimens were dissected to analyse the anatomy of the coracoclavicular ligament complex, the position of anchors and sutures and to measure the distance to the neurovascular structures at risk. The supraspinatus muscle was never injured by the Neviaser approach. The insertion of the suture anchors never failed, resulting in a secure and reproducible anchor position. The mean distance between the coracoid and suprascapular nerve was 1.8 cm (1.5-2.2), between the coracoid and the suprascapular artery 1.5 cm (1.3-1.9). These structures were never injured. The resulting force vector of the suture located between the anchor and the drill hole was close to the anatomic force vector of the coracoclavicular ligament complex. The suture cerclage was always correctly positioned. The presented technique is at minimal risk for the surrounding neurovascular structures and allows for a minimally invasive and anatomically correct reconstruction of the AC joint. Further biomechanical analysis is needed to evaluate the strength of the reconstruction technique. The proposed technique might be a reasonable alternative to existing invasive techniques of open reconstruction of acute type III AC-joint dislocations in high-demand patients. PMID- 16374589 TI - Arthroscopic biceps tendon tenodesis: the anchorage technical note. AB - Treatment of long head biceps (LHB) tendon pathology has become an area of renewed interest and debate among orthopaedic surgeons in recent years. The background of this manuscript is a description of biceps tenodesis which ensure continual dynamic action of the tendon which depresses the head and impedes lateral translation. A new technique has been developed in order to treat LHB tendon irreversible structural abnormalities associated with cuff rotator lesions. This technique entails the construction of a biological anchor between the LHB and supraspinatus and/or infraspinatus tendons according to arthroscopic findings. The rationale, although not supported by biomechanical studies is to obtain a triple, biomechanical effect. The first of these biomechanical effects which we try to promote through the procedure of transposition is the elimination of the deviation and oblique angle which occurs as the LHB completes its intra articular course prior to reaching the bicipital groove. Furthermore, we have found this technique extremely useful in the presence of large ruptures of the rotator cuff with muscle retraction. The most common complication associated to this particular method, observed in less than 3%, is failed biological fixation which manifests as subsidence of the tenodesis and consequent descent of the tendon with evident aesthetic deformity. PMID- 16374590 TI - Intra-articular pull out of an interference screw after anterior cruciate ligament reconstruction. AB - We report a rare case of late femoral interference screw migration in the posterior compartment of the knee after anterior cruciate ligament reconstruction. The graft was intact with no signs of damage. The screw was successfully removed through the posteromedial portal site and the patient regained full function of the knee. PMID- 16374591 TI - Cerebellar vermis morphology in children with spina bifida and Chiari type II malformation. AB - OBJECTIVE: Posterior fossa size and cerebellar weight and volume are reduced in Chiari type II malformation (CII). This is assumed to affect the cerebellum uniformly. We quantified the presumed reduction in vermis size on magnetic resonance imaging (MRI). METHODS: A midsagittal brain MRI slice was selected from each of 68 participants with CII (mean age 13 years). Control participants were 28 typically developing children (mean age 14.1 years). Midsagittal surface areas occupied by the intracranial fossa, posterior fossa, vermis, and its lobules were measured. CONCLUSIONS: Mean posterior fossa area was significantly smaller (P<0.003), although mean vermis area was significantly larger (P<0.0001), in participants with CII than in control participants. This expansion involved vermis lobules I-V and VI-VII areas (P<0.0001). The midsagittal vermis was expanded and not reduced in size in participants with CII. This is attributed to compressive displacement of midline structures within the confines of a small posterior fossa. PMID- 16374594 TI - Targeted mapping of ESTs linked to the adult plant resistance gene Lr46 in wheat using synteny with rice. AB - The gene Lr46 has provided slow-rusting resistance to leaf rust caused by Puccinia triticina in wheat (Triticum aestivum), which has remained durable for almost 30 years. Using linked markers and wheat deletion stocks, we located Lr46 in the deletion bin 1BL (0.84-0.89) comprising 5% of the 1BL arm. The distal part of chromosome 1BL of wheat is syntenic to chromosome 5L of rice. Wheat expressed sequence tags (ESTs) mapping in the terminal 15% of chromosome 1BL with significant homology to sequences from the terminal region of chromosome 5L of rice were chosen for sequence-tagged site (STS) primer design and were mapped physically and genetically. In addition, sequences from two rice bacterial artificial chromosome clones covering the targeted syntenic region were used to identify additional linked wheat ESTs. Fourteen new markers potentially linked to Lr46 were developed; eight were mapped in a segregating population. Markers flanking (2.2 cM proximal and 2.2 cM distal) and cosegregating with Lr46 were identified. The physical location of Lr46 was narrowed to a submicroscopic region between the breakpoints of deletion lines 1BL-13 [fraction length (FL)=0.89-1] and 1BL-10 (FL=0.89-3). We are now developing a high-resolution mapping population for the positional cloning of Lr46. PMID- 16374595 TI - Ultraweak photon emission from herbivory-injured maize plants. AB - Following perception of herbivory or infection, plants exhibit a wide range of inducible responses. In this study, we found ultraweak photon emissions from maize leaves damaged by Helicoverpa armigera (Noctuidae). Interestingly, mechanically damaged maize leaves treated with caterpillar regurgitants emitted the same intensity and pattern of photon emissions as those from maize leaves damaged by caterpillars. Furthermore, two-dimensional imaging of the leaf section treated with the oral secretions clearly shows that photon emissions were observed specifically at the lip of the wound exposed to the secretions. These results suggest that the direct interaction between maize leaf cells and chemicals contained in caterpillar regurgitants triggers these photon emissions. PMID- 16374593 TI - Spread of ampicillin/vancomycin-resistant Enterococcus faecium of the epidemic virulent clonal complex-17 carrying the genes esp and hyl in German hospitals. AB - The incidence of vancomycin-resistant Enterococcus faecium isolation was low (0.20). The relationship between plasma PLTP activity and IMT was not significant in control subjects. CONCLUSIONS/INTERPRETATION: Plasma PLTP activity is a positive determinant of IMT in type 2 diabetes mellitus, suggesting that high PLTP activity is involved in accelerated atherosclerosis in this disease. PMID- 16374628 TI - Plasma cytokine levels in young and elderly twins: genes versus environment and relation to in vivo insulin action. AB - AIMS/HYPOTHESIS: We studied the impact of genetic versus pre- and postnatal environmental factors on the plasma levels of IL6, TNF and the soluble TNF receptor superfamily, member 1A (TNFRSF1A, previously known as TNF receptor type1 [TNFR1]). MATERIALS AND METHODS: Using the hyperinsulinaemic-euglycaemic clamp, we assessed the association between cytokine levels and both peripheral insulin sensitivity and hepatic glucose production. Fasting plasma IL6, TNF and soluble TNFRSF1A levels were measured using ELISA in 55 young and 43 elderly twin pairs. RESULTS: Plasma IL6 and TNF were influenced by genetic factors in young and elderly twins respectively, while no significant impact of genetics was found for soluble TNFRSF1A. Significant intra-twin pair correlations between birthweight and both IL6 and soluble TNFRSF1A levels were found. In addition to the effect of birthweight on IL6, age and fat % also significantly influenced IL6 levels, whereas soluble TNFRSF1A levels were significantly influenced by zygosity, age, fat % and sex. Plasma soluble TNFRSF1A was associated with higher plasma NEFA and to some extent with reduced insulin sensitivity. Plasma IL6 was associated positively with basal NEFA. TNF level was not associated with in vivo glucose or fat metabolism after correction for known confounders. CONCLUSIONS/INTERPRETATION: Plasma IL6 and soluble TNFRSF1A are influenced by the intrauterine environment. We found some evidence of a genetic component for TNF and IL6 in young and elderly twins respectively. Plasma IL6 may influence basal lipolysis, but neither plasma TNF nor IL6 levels are independently associated with hepatic or peripheral insulin action. Nevertheless, plasma soluble TNFRSF1A may play some role in the control of insulin action. PMID- 16374629 TI - Effector-dependent and response location learning of probabilistic sequences in serial reaction time tasks. AB - We investigated the contributions of the sequences of effectors and response locations to probabilistic sequence learning in the serial reaction time task by means of bimanual transfer. Participants, trained with the dominant hand, were either required to maintain responding with the dominant hand after transfer or to switch to the nondominant hand. For both groups, half of the participants were transferred to the originally trained sequence, whereas the other half was transferred to a mirror-ordered sequence. This way, the sequence of effectors varied independently of the sequence of response locations. Sequence learning was assessed with probabilistic sequences, composed of either first-order or second order probabilities. Transfer of sequence knowledge to the nondominant hand was equally good for the originally trained sequence as for the mirrored sequence. This suggests that probabilistic sequence learning can be based on either the sequence of effectors or response locations. However, when participants responded with the dominant hand to a mirrored sequence, transfer performance was disturbed. This indicates that changing both the sequences of effectors and response locations has a disruptive effect on the learning performance. PMID- 16374630 TI - Functional asymmetry and interhemispheric cooperation in the perception of emotions from facial expressions. AB - The present study used the redundant target paradigm on healthy subjects to investigate functional hemispheric asymmetries and interhemispheric cooperation in the perception of emotions from faces. In Experiment 1 participants responded to checkerboards presented either unilaterally to the left (LVF) or right visual half field (RVF), or simultaneously to both hemifields (BVF), while performing a pointing task for the control of eye movements. As previously reported (Miniussi et al. in J Cogn Neurosci 10:216-230, 1998), redundant stimulation led to shorter latencies for stimulus detection (bilateral gain or redundant target effect, RTE) that exceeded the limit for a probabilistic interpretation, thereby validating the pointing procedure and supporting interhemispheric cooperation. In Experiment 2 the same pointing procedure was used in a go/no-go task requiring subjects to respond when seeing a target emotional expression (happy or fearful, counterbalanced between blocks). Faster reaction times to unilateral LVF than RVF emotions, regardless of valence, indicate that the perception of positive and negative emotional faces is lateralized toward the right hemisphere. Simultaneous presentation of two congruent emotional faces, either happy or fearful, produced an RTE that cannot be explained by probability summation and suggests interhemispheric cooperation and neural summation. No such effect was present with BVF incongruent facial expressions. In Experiment 3 we studied whether the RTE for emotional faces depends on the physical identity between BVF stimuli, and we set a second BVF congruent condition in which there was only emotional but not physical or gender identity between stimuli (i.e. two different faces expressing the same emotion). The RTE and interhemispheric cooperation were present also in this second BVF congruent condition. This shows that emotional congruency is the sufficient condition for the RTE to take place in the intact brain and that the cerebral hemispheres can interact in spite of physical differences between stimuli. PMID- 16374631 TI - Transsaccadic integration of visual features in a line intersection task. AB - Transsaccadic integration (TSI) refers to the perceptual integration of visual information collected across separate gaze fixations. Current theories of TSI disagree on whether it relies solely on visual algorithms or also uses extra retinal signals. We designed a task in which subjects had to rely on internal oculomotor signals to synthesize remembered stimulus features presented within separate fixations. Using a mouse-controlled pointer, subjects estimated the intersection point of two successively presented bars, in the dark, under two conditions: Saccade task (bars viewed in separate fixations) and Fixation task (bars viewed in one fixation). Small, but systematic biases were observed in both intersection tasks, including position-dependent vertical undershoots and order dependent horizontal biases. However, the magnitude of these errors was statistically indistinguishable in the Saccade and Fixation tasks. Moreover, part of the errors in the Saccade task were dependent on saccade metrics, showing that egocentric oculomotor signals were used to fuse remembered location and orientation features across saccades. We hypothesize that these extra-retinal signals are normally used to reduce the computational load of calculating visual correspondence between fixations. We further hypothesize that TSI may be implemented within dynamically updated recurrent feedback loops that interconnect a common eye-centered map in occipital cortex with both the "dorsal" and "ventral" streams of visual analysis. PMID- 16374632 TI - How performing a mental arithmetic task modify the regulation of centre of foot pressure displacements during bipedal quiet standing. AB - We investigated the effect of performing a mental arithmetic task with two levels of difficulty on the regulation of centre of foot pressure (COP) displacements during bipedal quiet standing in young healthy individuals. There was also a control condition in which no concurrent task was required. A space-time-domain analysis showed decreased COP displacements, along the antero-posterior axis, when participants concurrently performed the most difficult mental arithmetic task. Frequency-domain and stabilogram-diffusion analyses further suggested these decreased COP displacements to be associated with an increased stiffness and a reduction of the exploratory behaviours in the short term, respectively. PMID- 16374633 TI - Towards bacterial strains overproducing L-tryptophan and other aromatics by metabolic engineering. AB - The aromatic amino acids, L-tryptophan, L-phenylalanine, and L-tyrosine, can be manufactured by bacterial fermentation. Until recently, production efficiency of classical aromatic amino-acid-producing mutants had not yet reached a high level enough to make the fermentation method the most economic. With the introduction of recombinant DNA technology, it has become possible to apply more rational approaches to strain improvement. Many recent activities in this metabolic engineering have led to several effective approaches, which include modification of terminal pathways leading to removal of bottleneck or metabolic conversion, engineering of central carbon metabolism leading to increased supply of precursors, and transport engineering leading to reduced intracellular pool of the aromatic amino acids. In this review, advances in metabolic engineering for the production of the aromatic amino acids and useful aromatic intermediates are described with particular emphasis on two representative producer organisms, Corynebacterium glutamicum and Escherichia coli. PMID- 16374634 TI - Characterization of Streptomyces MITKK-103, a newly isolated actinomycin X2 producer. AB - A new actinomycete strain designated MITKK-103 was isolated from the soil of a flowerpot using a humic acid agar medium. The newly isolated strain was able to produce a large amount of actinomycin X2 even under nonoptimized growing conditions and serves as a promising source of this antibiotic. Actinomycin X2 has higher cytotoxicity toward cultured human leukemia (HL-60) cells than does actinomycin D, and it induces cell death via apoptosis. A nearly complete 16S ribosomal DNA (rDNA) sequence from the isolate was determined and found to have high identity (98.5-100%) with Streptomyces galbus, Streptomyces griseofuscus, and Streptomyces padanus, indicating that MITKK-103 belongs to the genus Streptomyces. The isolate clustered with species belonging to the S. padanus clade in a 16S-rDNA-based phylogenetic tree and showed 75% overall homology to S. padanus ATCC 25646 in DNA-DNA relatedness analysis. Although the growth of the isolate was somewhat different from the three species mentioned, the strain MITKK 103 most closely resembles S. padanus on the basis of the morphological and phenotypic characteristics, phylogenetic analysis, and genotypic data. As such, this is the first report of a strain of S. padanus capable of producing actinomycins. PMID- 16374635 TI - Hydrolysis of beta-1,3/1,6-glucan by glycoside hydrolase family 16 endo-1,3(4) beta-glucanase from the basidiomycete Phanerochaete chrysosporium. AB - When Phanerochaete chrysosporium was grown with laminarin (a beta-1,3/1,6-glucan) as the sole carbon source, a beta-1,3-glucanase with a molecular mass of 36 kDa was produced as a major extracellular protein. The cDNA encoding this enzyme was cloned, and the deduced amino acid sequence revealed that this enzyme belongs to glycoside hydrolase family 16; it was named Lam16A. Recombinant Lam16A, expressed in the methylotrophic yeast Pichia pastoris, randomly hydrolyzes linear beta-1,3 glucan, branched beta-1,3/1,6-glucan, and beta-1,3-1,4-glucan, suggesting that the enzyme is a typical endo-1,3(4)-beta-glucanase (EC 3.2.1.6) with broad substrate specificity for beta-1,3-glucans. When laminarin and lichenan were used as substrates, Lam16A produced 6-O-glucosyl-laminaritriose (beta-D-Glcp-(1->6) beta-D-Glcp-(1->3)-beta-D-Glcp-(1->3)-D-Glc) and 4-O-glucosyl-laminaribiose (beta D-Glcp-(1->4)-beta-D-Glcp-(1->3)-D-Glc), respectively, as one of the major products. These results suggested that the enzyme strictly recognizes beta-D-Glcp (1->3)-D-Glcp at subsites -2 and -1, whereas it permits 6-O-glucosyl substitution at subsite +1 and a beta-1,4-glucosidic linkage at the catalytic site. Consequently, Lam16A generates non-branched oligosaccharide from branched beta 1,3/1,6-glucan and, thus, may contribute to the effective degradation of such molecules in combination with other extracellular beta-1,3-glucanases. PMID- 16374636 TI - T-cell receptor tetramer binding or the lack there of does not necessitate antigen reactivity in T-cell receptor transduced T cells. AB - Genetic transfer of T-cell receptor (TCR) chains provides a means of transferring tumor antigen specificity onto an alternate T-cell population. To determine which tumor reactive TCRs are best suitable for such adoptive transfer, careful evaluation of the resulting TCR modified populations need to be performed. We have previously cloned, and expressed TCRs from melanoma, EBV, HCV, and HPV reactive T-cell clones and found that several routine indicators of T-cell function do not always predict the relative strength of a TCR. Using a combination of tetramer binding assays and antigen recognition assays, we identified TCRs that fall into three classes. One class of TCR did not bind tetramers yet resulted in cells with high avidity for antigen. A second TCR class bound tetramer but did not secrete cytokines in response to antigen. Finally, the third class of TCRs bound tetramer and reacted to antigen as would be expected. We conclude that tetramer binding is not always a good indicator of the function of a cloned TCR or the avidity of a TCR gene modified T cell. Furthermore, our data indicate that the use of tetramer binding alone to identify antigen reactive TCRs may result in the exclusion of TCRs that may be highly reactive or cross reactive to the relevant tumor antigen. PMID- 16374637 TI - Impact of the cyclosporine-ketoconazole interaction in children with steroid dependent idiopathic nephrotic syndrome. AB - BACKGROUND: Children with steroid-dependent nephrotic syndrome experience serious side effects from steroid therapy. Cyclosporine A (CsA), which is an effective agent in the treatment of steroid-dependent nephrotic syndrome, is expensive and, consequently, often unaffordable in developing countries. Many studies have documented the benefit of ketoconazole administration in transplant adults treated with CsA. We have conducted a retrospective study with the objective of addressing cost-savings, safety, and the efficacy of the co-administration of ketoconazole and CsA to children with steroid-dependent nephrotic syndrome. METHODS: This study included 102 nephrotic patients who were steroid-dependent and who received cyclosporine therapy. The commonest pathologic lesions were focal segmental glomerulosclerosis (64 patients) and minimal change disease (36 patients). Among the patients participating in the study, 78 received daily ketoconazole therapy (ketoconazole group) in the form of a 50-mg dose accompanied by an initial one-third decrease in the CsA dose, while 24 received CsA alone (non- ketoconazole group). All of the patients were children (below 18 years), and the male-to-female ratio was 3:1. The mean duration of treatment was 22.9 months. The characteristics of both groups were comparable. RESULTS: Co administration of ketoconazole significantly reduced mean doses of CsA by 48% with a net cost savings of 38%. It also resulted in a significant improvement in the CsA response and a more successful steroid withdrawal as well as a decrease in the frequency of renal impairment. Liver function tests remained normal in both groups up to and including the final follow-up (mean of 33.6 months). CONCLUSIONS: The co-administration of ketoconazole to CsA in children with idiopathic steroid-dependent nephrotic syndrome safely results in a significant reduction in CsA cost, which causes great concern in developing countries. It may also improve CsA response. PMID- 16374638 TI - Nucleotide regulation of paracellular Cl- permeability in natural rabbit airway epithelium. AB - In this study, we demonstrate a novel regulatory mechanism by which mucosal nucleotides via P2Y receptors decrease paracellular Cl(-) ion permeability in natural rabbit airway epithelium (in addition to a decrease in active Na(+) absorption). In contrast to primary cultures, the natural airway epithelium is a low-resistance epithelium, and an equivalent circuit model predicts that changes of more than approximately 12% in transepithelial conductance (G (t)) must include an effect on paracellular conductance (G (s)). Mucosal P2Y receptor stimulation with uridine triphosphate (UTP; 200 microM) decreased G (t) by up to 50% (average, 24%) and simultaneously decreased the paracellular Cl(-) permeability (mucosa-to-serosa Cl(-) flux) by 16%, but had no effect on mannitol permeability. The G (t) response to UTP was mimicked and attenuated by ionomycin (1 microM), suggesting a dependence on Ca(2+) (i). Amiloride (100 microM) and hyperosmolarity (+75 mM mannitol) also decreased G (t), indicating a role of cell shrinkage. Elevation of cAMP with forskolin (8 microM) or isoproterenol (10 microM) increased G (t) by 55 and 32%, and forskolin increased paracellular Cl(-) permeability by 37% without affecting mannitol permeability. The opposite effects of Ca(2+) (i) and cAMP on G (t) suggest an autocrine nucleotide signaling sequence where P2Y-dependent decrease in passive, paracellular Cl(-) transport is succeeded by a reversion of this effect due to P1-receptor-stimulated cAMP formation by adenosine originating from a time-dependent breakdown of mucosal ATP. PMID- 16374639 TI - Hair-dye allergy: a coloured case. PMID- 16374640 TI - [Therapy of arthrofibrosis after total knee arthroplasty]. AB - Arthrofibrosis is one of the most common complications after total knee arthroplasty with an overall incidence of approximately 10%. Nevertheless, published data are rare and clinical trials mostly include small and heterogeneous patient series resulting in controversial conclusions. Clinically, arthrofibrosis after knee arthroplasty is defined as (painful) stiffness with scarring and soft tissue proliferation. Differentiation between local (peripatellar) and generalized fibrosis is therapeutically relevant. Histopathology typically shows subsynovial fibrosis with synovial hyperplasia, chronic inflammatory infiltration, and excessive and unregulated proliferation of collagen and fibroblasts. Diagnostic strategies are based on the exclusion of differential causes for painful knee stiffness, and especially the exclusion of low-grade infections represents a diagnostic challenge. Early and intensive physiotherapy combined with sufficient analgesia should be initiated as a basic therapy. The next therapeutic steps for persisting arthrofibrosis include closed manipulation and open arthrolysis. Arthroscopic interventions should be limited to local fibrosis. Revision arthroplasty represents a rescue surgery, often associated with recurrence of fibrosis. Prevention of arthrofibrosis by sufficient analgesia and early physiotherapy remains the best treatment option for painful stiffness after knee arthroplasty. PMID- 16374641 TI - Preface: The invitation of the CERC3 to organize a "Young Chemists' Workshop". PMID- 16374642 TI - [The development of evidence-based psychotherapy for use in alcoholism. A review]. AB - Past decades have seen enormous advances in the development and validation of evidence-based approaches to psychotherapy for alcoholism. While psychoanalytic and early behavioral techniques were the original basis of psychotherapy in this field, evidence-based approaches are now built up on the principles of motivational interviewing and cognitive-behavior therapy. Different techniques have been developed to modify preceding and persisting conditions favoring problem behavior, e.g., training in coping/social skills and the community reinforcement approach. According to the results of the project MATCH, one of the largest treatment trials, "cognitive-behavioral intervention" combines motivational enhancement therapy, the 12-step approach, and cognitive-behavior therapy, with the aim of providing new and even more efficacious psychotherapy for alcohol dependent patients. These very promising developments are beset with huge problems, however, insofar as few of the new evidence-based treatment approaches are accepted as standard treatment in Germany, in addition to which only a fraction of all alcohol-dependent persons in the country find their way into the care system, for various reasons. Early diagnosis and facilitation of access to the various treatment options available could be a future task for general practitioners and also for company / industrial medical schemes. PMID- 16374643 TI - Stabilisation of periprosthetic fractures with angular stable internal fixation: a report of 13 cases. AB - INTRODUCTION: Periprosthetic fractures of the femur present a challenging surgical problem. The aim of this study was to retrospectively evaluate the outcome of periprosthetic fractures stabilised with an angular stable, less invasive stabilisation system (LISS). PATIENTS AND METHODS: Thirteen patients (ten total hip-, two total knee-, one total hip- and knee-arthroplasty) with periprosthetic fractures were treated with the LISS internal fixator (in ten cases minimal invasive). Six patients had previous operations due to periprosthetic fractures. The average follow-up period was 20 months, follow-up rate 85%. RESULTS: All fractures showed radiographic fracture healing without implant loosening. Except one patient, all patients had returned to their pre operative activity level. No early post-operative complications were seen. There was one implant failure after 4 months and two cases of malunion. CONCLUSION: The cases showed the internal fixator to be effective for the stabilisation of periprosthetic fractures, even in cases of poor bone quality with good functional outcomes. The internal fixator, with the option of minimal invasive application, is the preferred method of osteosynthesis in periprosthetic fractures. PMID- 16374644 TI - In vitro gene targeting in human hepatoblastoma. AB - Poor treatment results in advanced hepatoblastoma (HB) made alternative treatment approaches desirable. Gene-directed tumor therapy is increasingly investigated in different malignancies. The aim of this study was to analyze possible alternatives of gene transfer into HB cells and to study therapeutic applications based on different strategies. Liposomal transfection of HB cells was assessed using liver-specific promoters, and adenovirus and Sendai virus transductions were performed in vitro. Transfer efficiencies were measured via flow cytometry determining expression of vector-encoded marker gene green fluorescent protein. Gene silencing of the anti-apoptotic bcl-2 gene in HUH6 cells was performed using lipofection of small interfering RNA (siRNA). Additionally, suicide gene therapy was carried out through a yeast-derived cytosine deaminase (YCD)-combined yeast uracil phosphoribosyltransferase (YUPRT)-based adenovirus-mediated gene transfer, leading to a potent intracellular prodrug transformation of 5-fluorocytosine into 5-fluorouracil. Treatment efficiencies were monitored via MTT viability assay. Highest gene transfer rates (86%) were observed using adenovirus transduction. We furthermore observed a significant therapeutic effect of adenovirus-mediated YCD::YUPRT suicide gene transfer. Liposomal-mediated anti-bcl-2 siRNA transfer led to a significant improvement of cisplatin treatment in HUH6 cells. Liver specific promoters were found to be strongly active in HUH6 cells (mixed HB derived), but less active in HepT1 cells (embryonal HB-derived). Liposomal transfection and viral transduction are effective approaches to genetically manipulate HB cells in vitro. For the first time, we demonstrate a positive effect of siRNA gene silencing in this malignancy. Additionally, we successfully investigated a model of adenovirus-based suicide gene therapy in HB cell cultures. Our data strongly encourage further studies assessing these alternative treatment approaches. PMID- 16374645 TI - Transcatheter arterial embolization in a hemodynamically unstable patient with grade IV blunt liver injury: is nonsurgical management an option? AB - The prevalence of liver injury in patients who have sustained blunt multiple trauma was reported to range from 1 to 8%. Because previous mortality rates were as high as 50-80% for severe hepatic injury, the choice of treatment was under intensive investigation. Whereas nonsurgical management was the standard treatment for the hemodynamically stable patient, there is no consensus on how to treat hemodynamically unstable patients. This report details the case of a patient who sustained blunt multiple trauma, resulting in a grade IV liver injury, graded according to the American Association for the Surgery of Trauma (AAST) Liver Injury Scale. With massive fluid and blood resuscitation, the patient was stable enough to be managed nonsurgically. With transcatheter arterial embolization (TAE), the left and right hepatic arteries were embolized with coils, which allowed for a good recovery. We hypothesize that TAE can be used in the hemodynamically unstable patient who responds to rapid fluid resuscitation and blood transfusion. We caution that there is insufficient evidence until now and would therefore not make any recommendations; however, we would question the need for surgery in unstable patients with this kind of injury in the future. PMID- 16374646 TI - The incidence and distribution of burst fractures. AB - This study was conducted to assess occurrence and distribution of burst fractures as well as common accident mechanisms and the associated neurologic deficit. Using picture archiving and communications system, all emergency multidetector computed tomography studies over 34 months were retrieved and evaluated for burst fractures by two radiologists by consensus. Initial neurological findings were retrieved from patients' medical records. One hundred fifty-two patients (112 male) with a total of 169 burst fractures were found. In both genders, the incidence of burst fractures peaked at the thoracolumbar junction and between levels T5 and T8. In 10% of cases, more than one burst fracture was seen, thereof 53% on noncontiguous levels. Main accident mechanisms were falls, traffic, and sports. Neurological deficit was highest in patients with burst fractures of the cervical spine, independent of accident mechanism, and lowest in thoracolumbar junction fractures. Burst fractures occur frequently in high energy traumas and are most commonly associated with falling and traffic accidents. Multiple burst fractures occur in 10% of cases, half thereof on noncontiguous levels. PMID- 16374647 TI - Frameless image-guided neuronavigation in orbital surgery: practical applications. AB - Frameless neuronavigation has been increasingly used in intracranial and spinal neurosurgery. However, the application of neuronavigation in orbital surgery has not yet been reported. The purpose of this study was to evaluate whether frameless navigation in the orbit provides sufficient clinical application accuracy and thus a useful tool in orbital surgery. A frameless infrared-based neuronavigation system (VectorVision, BrainLAB, Heimstetten, Germany) was used in the microsurgical removal of 11 orbital tumors (five cavernomas, three lymphomas, two dermoid tumors, and one rhabdomyosarcoma). The transconjunctival approach was performed in seven cases, lateral orbitotomy in three cases, and the extradural pterional approach was performed in one case. The surgery was successful and without any complications in all cases. The registration accuracy of the neuronavigation was between 1.8 and 2.2 mm, with a mean of 1.9 mm. Subtotal tumor debulking was performed in one case with lymphoma. One patient was only biopsied due to suspected systemic lymphoma disease. Total removal of the tumor could be accomplished in the other nine patients. Image guidance during orbital surgery offered excellent three-dimensional guidance on the surface of the intraorbital lesions, allowing a safer, more controlled surgery. The surgical targets in the orbit are fixed structures, thus no shifting occurs and continuous high intraoperative navigation accuracy can be achieved. The use of the navigation clearly reduces the operative risk and increases the effectiveness of microsurgical orbital procedures. PMID- 16374648 TI - A laboratory training model for interhemispheric-transcallosal approach to the lateral ventricle. AB - Laboratory training models are essential for developing and refining surgical skills before clinical application of microneurosurgery. Our aim is to train residents of neurosurgery to be familiar with a basic microneurosurgical technique in access to the lateral ventricle via a transcallosal approach. The training material consists of a 2-year-old fresh cadaveric cow cranium. A four step approach was designed to simulate microneurosurgical dissection along the falx to visualize cingulated gyri, callosomarginal and pericallosal arteries in order to perform callosotomy and access to the lateral ventricle, and finally to the foramen of Monroe. We conclude that the model perfectly simulates standard microneurosurgical steps in interhemispheric-transcallosal approach to the lateral ventricle and to the area of the foramen of Monroe. PMID- 16374649 TI - Outcome predictors and complications in the management of intradural spinal tumours. AB - The results of the management of 115 patients with intradural spinal tumours are presented. Data was collected retrospectively from the case notes. Tumours were categorized as intramedullary or extramedullary for statistical analysis. Meningioma, schwannoma and ependymoma accounted for 70% of tumours. Complete macroscopic excision was achieved in 84% of extramedullary and 54% of intramedullary tumours. There were two post-operative deaths, one of which was secondary to methacillin-resistant staphylococcus aureus (MRSA) meningitis. Cerebrospinal fluid leak (10%) and meningitis (7%) were the commonest complications. Ninety-six percent of patients with extramedullary tumours improved or remained unchanged on the Frankel scale. In the intramedullary group, 82% remained unchanged or improved after treatment. Pre-operative functional status was a predictor of good post-operative function for intra- and extramedullary tumours and for intramedullary tumours a good post-operative Frankel score predicted long-term survival. PMID- 16374650 TI - Morphologic comparison of cervical, thoracic, lumbar intervertebral discs of cynomolgus monkey (Macaca fascicularis). AB - The aim was to analyze the morphological differences of the intervertebral disc and endplates at different levels. Forty-five vertebral motion segments were obtained from the spine of nine 3 to 4-year-old cynomolgus monkeys (Macaca fascicularis). From every spine, five discs were sectioned (C5-C6, T3-T4, T9-T10, L2-L3, L4-L5). In all the groups, tissue samples were collected and sections were stained with Masson's trichrome, Safranine-O and van Gieson's connective tissue stain to analyze the intervertebral discs. Immunohistochemistry was performed, using specific antibodies to detect collagens I and II. The intervertebral disc height, the maximum nucleus pulposus height, the superior and inferior endplate heights were histomorphometrically measured and different indexes were calculated to compare the differences between specimens of the same animal and between discs of the same level, and finally the differences between groups of discs of different levels. There were no differences existing in annular fibers anchoring on the endplate between discs of different levels. A positive immune reaction for type I collagen was observed in the longitudinal ligaments and in the annular region adjacent to them. Collagen II immune reactivity was found in the annulus close to the nucleus pulposus, in the endplates and in the nucleus. There were no differences between discs of different levels in the collagen I and II localization. The height of the discs varied along the spine. The smallest value was measured in T3-T4, with a larger increase caudally than cranially. The highest value was measured in L2-L3. A cervical disc was 55% the height of a lumbar one. The endplate height increased along the length of the spine. The inferior EP was always higher than the superior. The study provides a detailed structural characterization of the intervertebral disc and may be useful for further investigations on the disc degeneration process. PMID- 16374651 TI - The total costs of a displaced femoral neck fracture: comparison of internal fixation and total hip replacement. A randomised study of 146 hips. AB - We randomised 143 patients--age 75 years or older--with displaced femoral neck fracture to either internal fixation or total hip replacement (THR) and compared the socio-economic consequences. In the internal fixation group, 34 of 78 hips underwent secondary surgery. In the THR group, 12 of 68 hips dislocated, the majority in mentally impaired patients. We calculated the total hospital costs for two years after operation. When secondary surgery was included, there was no difference in costs between the internal fixation and THR groups, or between the mentally impaired and lucid subgroups. The costs to the community were calculated comparing the baseline cost before surgery with the average cost per month during the first postoperative year. No difference was found between the treatment groups. The Harris hip scores were higher in the THR group, and pain was more common in the internal fixation group. In lucid patients, THR gives a better clinical result at the same cost. PMID- 16374652 TI - The use of levo-carnitine in children with renal disease: a review and a call for future studies. AB - Carnitine is an amino acid derivative that has a key role in the regulation of fatty acid metabolism and ATP formation. Carnitine deficiency has been described in various conditions, including chronic kidney disease (CKD) and end stage renal disease (ESRD). The deficiency of this micronutrient is postulated to lead to adverse effects across multiple organ systems. There is a paucity of information on carnitine deficiency and its effects in the pediatric CKD and ESRD populations. Currently, there is no evidence supporting the routine use of carnitine supplementation in children with ESRD. In this article, we review the pathophysiology, pharmacokinetics and the potential effects of levo-carnitine supplementation with a focus on the pediatric CKD and ESRD populations. Finally, potential future directions of research are discussed. PMID- 16374653 TI - Trigemino-cardiac reflex during skull base surgery: a new entity of ischaemic preconditioning? The potential role of imaging. PMID- 16374655 TI - Microwave-accelerated metal-enhanced fluorescence (MAMEF): application to ultra fast and sensitive clinical assays. AB - In this rapid communication we describe an exciting platform technology that promises to fundamentally address two underlying constraints of modern assays and immunoassays, namely sensitivity and rapidity. By combining the use of Metal enhanced Fluorescence (MEF) with low power microwave heating (Mw), we can significantly increase the sensitivity of surface assays as well as >95% kinetically complete the assay within a few seconds. This technology is subsequently likely to find significant importance in certain clinical assays, such as in the clinical assessment of myoglobin, where both the assay rapidity and sensitivity are paramount for the assessment and treatment of acute myocardial infarction. PMID- 16374656 TI - Gambling and the older Chinese in Canada. AB - In Canada, there is a lack of research on gambling among the older adults from ethnic minority groups, especially the older Chinese. In this study, two research questions were used to examine gambling among the older Chinese: (1) What is the pattern of gambling among the older Chinese in Canada? (2) What are the predictors associated with gambling among the older Chinese in Canada? The data for this study were collected as part of a multi-site study on health and well being of 2272 older Chinese in Canada. Four main questions related to gambling were used in this study. Among the 2257 participants who answered the questions on gambling, 26.6% of them reported that they gambled. Results of the hierarchical logistic regression analysis showed that being male, having lived in Canada longer, having a higher level of social support, having more service barriers, and having a stronger level of Chinese ethnic identity would increase the probability for an older Chinese to participate in gambling. Conversely, having a post-secondary and above level of education and having a higher level of life satisfaction would reduce one's probability of gambling. Although city of residency was also significant in predicting gambling, further analysis showed that its effect was actually caused by other factors including services barriers, social support, life satisfaction, Chinese ethnic identity, and education. PMID- 16374657 TI - The four Es of problem gambling: a psychological measure of risk. AB - A focus group of Reno area Gamblers Anonymous members identified four psychological traits contributing to risk for problem gambling, including: Escape, Esteem, Excess and Excitement. A panel of four experts authored 240 Likert-type items to measure these traits. By design, none of the items explicitly referred to gambling activities. Study 1 narrowed the field of useful items by employing a quasi-experimental design which compared the answers of Reno area Gamblers Anonymous members (N = 39) to a control sample (N = 34). Study 2 submitted successful items, plus new items authored with the knowledge gained from Study 1, to validation in a random sample telephone survey across Queensland, Australia (N=2577). The final 40 item Four Es scale (4Es) was reliable (alpha=.90); predicted gambling problems as measured by the Canadian Problem Gambling Index of Severity (PGSI, Ferris & Wynne (2001). The Canadian Problem Gambling Index: Final Report: Canadian Centre on Substance Abuse); and distinguished problem gamblers from persons with alcohol abuse problems. The new scale can provide a basis for further study in harm minimization, treatment, and theory development. PMID- 16374658 TI - Characteristics and gender differences among self-excluded casino problem gamblers: Missouri data. AB - The present study explores gender-related differences in the demographic and gambling-related characteristics of 2670 problem gamblers participating in a state-administered (Missouri) casino self-exclusion program between 2001 and 2003. Female (n=1298, 48.4%) and male (n=1372, 51.1%) participants ranged in age from 21 to 84 years. Gender-related differences were noted among demographic variables, patterns of gambling behavior, reasons for self-exclusion, and involvement in self-help, counseling, and bankruptcy services. Female self excluders were more likely than males to be older at time of application, African American, and either retired, unemployed or otherwise outside the traditional workforce. In addition, female self-excluders were more likely to report a later age of gambling onset, a shorter period between onset and self-exclusion, a preference for non-strategic forms of gambling and prior bankruptcy. The main predictors for female participation in self-exclusion included a desire to gain control and prevent suicide and referral by a counselor. The desire to save the marriage was a motivating factor for all participants. Findings suggest that the most efficacious treatment strategies with this group will include family systemic therapy and financial management in addition to pharmaco-treatment and culturally-sensitive individual therapy. PMID- 16374659 TI - Men & women playing games: gender and the gambling preferences of Iowa gambling treatment program participants. AB - Historically, gambling has been a predominantly male pastime; however, as legalized gambling has expanded, female participation has increased. Nevertheless, some research suggests that a divide remains between the play patterns of men and women. For example, research suggests that men gravitate towards casino table games and track betting and women are attracted to games such as bingo and casino slots. Researchers have hypothesized that play pattern disparities exist because of inherent differences between the natures of men and women. Using data from 2256 (1309 male) problem gambling treatment participants, this research examines the influence of gender on play patterns. We tested the ability of gender and a series of demographic, economic, and health-related factors to discriminate among three groups of gamblers with different game preferences: casino preferred, slots preferred, and non-institutional preferred. The results of multiple discriminant function analyses indicated that gender provided a minimal contribution to discrimination beyond that of specific demographic, economic, and health-related factors. This finding suggests that for understanding gambling patterns, gender is less informative than descriptive gambler profiles. PMID- 16374660 TI - A gender-based examination of past-year recreational gamblers. AB - BACKGROUND: Most adults gamble recreationally yet few studies have systematically investigated for gender-related differences in recreational gamblers. METHODS: Logistic regression analyses were performed on data from a nationally representative sample of respondents from the 1998 Gambling Impact and Behavior Study. RESULTS: Female gamblers versus non-gamblers were more likely to report use of alcohol and drugs. Male gamblers versus non-gamblers were more likely to report alcohol use and abuse/dependence, any substance abuse/dependence, and lifetime incarceration. An analysis of gambling by gender group interaction effects did not identify significant differences in health-related measures. Male as compared with female gamblers reported beginning gambling earlier, experiencing higher past-year maximal losses and wins, and finding favorite and engaging in different types of gambling. CONCLUSIONS: Despite heavier gambling in male as compared with female recreational gamblers, similar mental health functioning was observed in female and male past-year recreational gamblers. Types of gambling problematic for men and women are reflected in the gambling preferences of recreational gamblers. PMID- 16374661 TI - Prevalence estimates of gambling participation and problem gambling among 16-18 year-old students in Iceland: a comparison of the SOGS-RA and DSM-IV-MR-J. AB - The aim of this study was to estimate gambling participation and problem gambling among Icelandic adolescents. Participants were 750 16-18-year-old students, 371 girls and 379 boys. The rate of problem gambling was estimated with the SOGS-RA and DSM-IV-MR-J. Results indicated that 96% of adolescents had gambled in their lifetime, 79% at least once in the preceding year and about 10% gamble at least once a week. A psychometric evaluation of the two screening scales revealed satisfactory reliabilities and factor structures for both scales. The DSM-IV-MR-J identified 2% of the participants as problem gamblers while SOGS-RA identified 2.7%, and problem gambling was more common among boys than girls. It was concluded that problem gambling among adolescents is an area of concern for the Icelandic community that needs to be further investigated. PMID- 16374662 TI - Population dynamics of cotton rats (Sigmodon hispidus) inhabiting abandoned petroleum landfarms in Oklahoma, USA. AB - Previous work by our group has shown bioaccumulation of contaminants and alterations in the immune system of hispid cotton rats (Sigmodon hispidus) inhabiting petrochemical waste sites (landfarms). We studied populations of cotton rats inhabiting petrochemical landfarms or uncontaminated reference sites. Cotton rat populations inhabiting abandoned landfarms experienced reduced summer population densities and lower mean monthly survival, with maximum densities 65% that of populations inhabiting non-contaminated grassland (reference) sites. Survival was lower in populations from landfarms (0.62+/-0.04) compared to reference sites (0.75+/-0.04), with differences most notable during summer months. Cotton rat populations sampled from landfarms had even sex ratios and a lower proportion of juveniles compared to populations from reference sites. No differences were observed in the weight of epididymides, testes, uteri, or ovaries, and no differences in fecundity were detected among sites. PMID- 16374663 TI - Water quality assessment by ecotoxicological and limnological methods in water supplies, southeast Brazil. AB - The quality of the water supplied to Araraquara city (Sao Paulo State, Brazil) was analyzed at four points: two reservoirs from which water is pumped and the water incoming and exiting a drinking-water treatment plant. Physical and chemical analyses of water were performed, as well as acute and chronic toxicity tests of the water from all four points, using Ceriodaphnia silvestrii (Cladocera, Crustacea) as test-organism. The study was carried out monthly in the period between December 2000 and November 2001. Water from the reservoirs caused neither acute nor chronic toxicity to test-organisms. On the contrary, it promoted an increase in the fecundity of cladocerans compared to control, suggesting greater availability of food. However, the water from exiting the treatment plant did cause chronic toxicity to C. silvestrii, regarding both fecundity and mortality. The toxicity was probably caused by residual chlorine, perhaps not completely removed by aeration, or by chlorine disinfection by products (trihalomethanes) in the treated water, considering the level of organic matter present in the reservoir water. PMID- 16374664 TI - Comparison of active venom components between Eastern brown snakes collected from South Australia and Queensland. AB - The abundance and activity of the prothrombin activator (pseutarin C) within the venom of the Eastern brown snake (Pseudonaja textilis textilis) is the primary determinant of its coagulation potency. Textilinin-1, also in this venom, is a plasmin inhibitor which is thought to exert its toxic effects through the slowing of fibrinolysis. The aim of this report is to determine if there are differences in the potency of the venom from Eastern brown snakes collected from South Australia (SA) compared to those from Queensland (QLD). A concentration of 0.4 microg/ml venom protein from six QLD specimens clotted citrated plasma in an average time of 21.4+/-3.3 s compared to 68.7+/-2.4 s for the same amount of SA venom (averaged for six individuals). The more potent procoagulant activity of the QLD venom was measured between 0.4 and 94 microg/ml venom protein in plasma. The anti-plasmin activity of textilinin was also greater in the venom of the snakes collected from QLD, causing full inhibition of plasmin at approximately 1.88 microg/ml of venom protein compared to approximately 7.5 microg/ml for the SA venoms. It is concluded that geographic differentiation of the Eastern brown snakes results in significant differences venom potency. PMID- 16374665 TI - Perylene toxicity in the estuarine environment of Ria de Aveiro (Portugal). AB - Perylene, a 5-ring polycyclic aromatic hydrocarbon is common in estuarine sediments and its toxicity in the benthic and planktonic compartments is not yet clarified. The objectives of this work were: (1) to follow the toxicity of high concentrations of perylene (110 mg l(-1)) on benthic bacteria and macrofauna (amphipod Corophium multisetosum); (2) to determine the effects of a low load of perylene (2 microg l(-1)) on the metabolism of suspended bacteriobenthos after 9 day exposure, mimicking the effects of tidal erosion; (3) to contrast the effects of this low perylene load on the particle-free bacterioplankton and on the suspended and particle-adhered bacteriobenthos. No impact was detected in bacterial abundance exposed to 110 mg perylene l(-1) for 9 days. This concentration of perylene evoked no acute effects in C. multisetosum but, chronic toxicity assays revealed statistically significant negative effects on survival, growth and number of pregnant females. The bacterioplankton and the suspended bacteriobenthos, exposed to 2 microg perylene l(-1) during 2 weeks, responded with altered profiles of activity when compared to the control suspension. These values ranged, respectively, for bacterial biomass production from 134 to 210 and from 24 to 184 mug C l(-1) h(-1), for aminopeptidase from 1824 to 11,127 and from 1464 to 15,488 nmol l(-1) h(-1), and for beta-glucosidase from 87 to 400 and from 57 to 1278 nmol l(-1) h(-1). The rate of oxygen consumption in the perylene exposed suspension (0.04-2.85 mmol O2 kg(-1) dw sed h(-1)) exhibited a clearly distinct profile in relation to the control (0.57-1.60 mmol O2 kg(-1) dw sed h( 1)). The overall reactivity of the bacteriobenthos to perylene was interpreted as the result of toxic pressure followed by evolution of a diverse bacterial community. PMID- 16374666 TI - Monitoring the effects of water pollution on Cyprinus carpio in Karakaya Dam Lake, Turkey. AB - Karakaya Dam Lake (KDL) is one of the most important water sources, both for irrigation and fishery, located in eastern part of Turkey. This study is concerned with the pollution of the lake contributed by urban, industrial and agricultural activities. The parameters selected for this aim were the enzymes commonly used as biomarkers of environmental pollution. The activity of glutathione S-transferase (GST), carboxylesterase (CE), lactate dehydrogenase (LDH), acid phosphatase (ACP) and aspartate amino transferase (AST) has been determined in liver tissue samples of Cyprinus carpio, a representative species of KDL. Furthermore, brain acetylcholinesterase (AChE) activity which is mainly affected by pesticides such as organophosphates, has been assayed. Chemical analysis results showed that KDL was polluted by various heavy metals as it was apparent from water, sediment and gill tissue. The activity of brain AChE was significantly lower in all localities than Tecimli area (St-5) where there is no agricultural and industrial activities in the immediate periphery. Thus, this change of AChE activity may relate to agricultural pollution in KDL. On the other hand, no significant differences were found for selected enzyme biomarkers, but condition factor (CF) or hepatosomatic index were significantly different from the St-5 samples, a result that may be attributed to water pollution in KDL by various contaminants. PMID- 16374667 TI - Bisphenol A in artificial indoor streams: II. Stress response and gonad histology in Gammarus fossarum (Amphipoda). AB - The effects of the world wide-distributed chemical bisphenol A (BPA) on the endocrine system of vertebrates have been demonstrated in several studies. Here, we report on the impact of BPA (0, 5, 50 and 500 microg/l nominally, deduced effective concentrations 0, 0.24, 2.4, and 24.1 microg/l, respectively, all at 15 degrees C) on the 70 kD stress protein family (hsp70), the 90 kD stress protein family (hsp90), and gonad histology of the crustacean Gammarus fossarum exposed in artificial indoor streams. The animals were exposed for a maximum of 103 days and samples were taken at the beginning and at days 34, 69 and 103 of the experiment. Exposure to BPA resulted in accelerated maturation of oocytes in females and in a decline in the number and size of early vitellogenic oocytes. The level of hsp90, which plays a pivotal role in vertebrate sex steroid signal transduction, was significantly reduced by BPA. In all five streams, measured parameters did not indicate any captivity stress for a period of 69 days. Beyond this time, the mortality rate and proteotoxic effects, the latter measured by hsp70 expression, were found to be elevated. PMID- 16374668 TI - HIV/STD stigmatization fears as health-seeking barriers in China. AB - Internationally, stigma prohibits effective HIV/STD identification, prevention, and care. Interviews with 106 persons in an urban center in Eastern China, some known to have engaged in stigmatized risk acts (sex workers, STD clinic patients) and some vulnerable for stigmatization fears to influence health-seeking behaviors (market employees, rural-to-urban migrants). Interviews focused on community norms, values, beliefs, and emotional and behavioral reactions to HIV/STD stigmatization related events. Attributions for infection were found to mark individual's failure to adhere to sexuality norms; define a condition warranting the avoidance of infected persons and dismissal by medical professionals; and promote anticipation of negative emotions (i.e., shame, fear, and embarrassment) and devalued social roles and status. Strategies reported to avoid stigmatization include avoiding HIV/STD knowledge; avoiding health care professionals, particularly in public settings; and conforming to community norms of shunning those suspected of risky behaviors. Results have direct implications for community marketing campaigns in China. PMID- 16374669 TI - Demonstrations of decreased sensitivity to complex motion information not enough to propose an autism-specific neural etiology. AB - Interest regarding neural information processing in autism is growing because atypical perceptual abilities are a characteristic feature of persons with autism. Central to our review is how characteristic perceptual abilities, referred to as perceptual signatures, can be used to suggest a neural etiology that is specific to autism. We review evidence from studies assessing both motion and form perception and how the resulting perceptual signatures are interpreted within the context of two main hypotheses regarding information processing in autism: the pathway- and complexity-specific hypotheses. We present evidence suggesting that an autism-specific neural etiology based on perceptual abilities can only be made when particular experimental paradigms are used, and that such an etiology is most congruent with the complexity-specific hypothesis. PMID- 16374670 TI - ERPs and eye movements reflect atypical visual perception in pervasive developmental disorder. AB - Many studies of eye tracking or event-related brain potentials (ERPs) in subjects with Pervasive Developmental Disorder (PDD) have yielded inconsistent results on attentional processing. However, recent studies have indicated that there are specific abnormalities in early processing that are probably related to perception. ERP amplitudes in response to visual stimuli, measured above the occipital (modality-specific) cortex, are reported to be abnormally small in patients with PDD, and the abnormal visual processing is possibly associated with the spatial visual frequency content of stimuli. It is suggested that subjects with PDD show abnormal activation of visual pathways dedicated to the processing of high and low spatial frequencies. PMID- 16374671 TI - Expectations and placebo response: a laboratory investigation into the role of somatic focus. AB - It has been theorized that expectations are an important causal determinant of the placebo effect. Placebo expectations, however, do not always yield placebo effects. In a laboratory study, we tested the hypothesis that one's level of somatic focus moderates the effect of placebo expectations on placebo responding. We also varied whether participants were told the placebo was a drug, could either be a drug or placebo, or was a placebo. The results revealed that individuals who thought they were taking a drug (i.e., unconditional expectations) reported more placebo symptoms when they closely focused on their symptoms. Individuals told they may or may not be receiving a drug (i.e., conditional expectations) did not differ from control participants regardless of how closely they attended to their symptoms. The findings have theoretical implications for expectancy models of the placebo effect as well as for practical research comparing the type of expectations held by individuals in clinical trials and clinical practice. PMID- 16374672 TI - Structure-activity relationships for a new family of sulfonylurea herbicides. AB - Acetohydroxyacid synthase (AHAS; EC 2.2.1.6) catalyzes the first common step in branched-chain amino acid biosynthesis. The enzyme is inhibited by several chemical classes of compounds and this inhibition is the basis of action of the sulfonylurea and imidazolinone herbicides. The commercial sulfonylureas contain a pyrimidine or a triazine ring that is substituted at both meta positions, thus obeying the initial rules proposed by Levitt. Here we assess the activity of 69 monosubstituted sulfonylurea analogs and related compounds as inhibitors of pure recombinant Arabidopsis thaliana AHAS and show that disubstitution is not absolutely essential as exemplified by our novel herbicide, monosulfuron (2-nitro N-(4'-methyl-pyrimidin-2'-yl) phenyl-sulfonylurea), which has a pyrimidine ring with a single meta substituent. A subset of these compounds was tested for herbicidal activity and it was shown that their effect in vivo correlates well with their potency in vitro as AHAS inhibitors. Three-dimensional quantitative structure-activity relationships were developed using comparative molecular field analysis and comparative molecular similarity indices analysis. For the latter, the best result was obtained when steric, electrostatic, hydrophobic and H-bond acceptor factors were taken into consideration. The resulting fields were mapped on to the published crystal structure of the yeast enzyme and it was shown that the steric and hydrophobic fields are in good agreement with sulfonylurea-AHAS interaction geometry. PMID- 16374673 TI - Genetic neural network modeling of the selective inhibition of the intermediate conductance Ca2+ -activated K+ channel by some triarylmethanes using topological charge indexes descriptors. AB - Selective inhibition of the intermediate-conductance Ca(2+)-activated K(+ )channel (IK (Ca)) by some clotrimazole analogs has been successfully modeled using topological charge indexes (TCI) and genetic neural networks (GNNs). A neural network monitoring scheme evidenced a highly non-linear dependence between the IK (Ca) blocking activity and TCI descriptors. Suitable subsets of descriptors were selected by means of genetic algorithm. Bayesian regularization was implemented in the network training function with the aim of assuring good generalization qualities to the predictors. GNNs were able to yield a reliable predictor that explained about 97% data variance with good predictive ability. On the contrary, the best multivariate linear equation with descriptors selected by linear genetic search, only explained about 60%. In spite of when using the descriptors from the linear equations to train neural networks yielded higher fitted models, such networks were very unstable and had relative low predictive ability. However, the best GNN BRANN 2 had a Q ( 2 ) of LOO of cross-validation equal to 0.901 and at the same time exhibited outstanding stability when calculating 80 randomly constructed training/test sets partitions. Our model suggested that structural fragments of size three and seven have relevant influence on the inhibitory potency of the studied IK (Ca) channel blockers. Furthermore, inhibitors were well distributed regarding its activity levels in a Kohonen self-organizing map (KSOM) built using the inputs of the best neural network predictor. PMID- 16374674 TI - Long-term outcome after laparoscopic and open surgery for rectal prolapse: a case control study. AB - BACKGROUND: Laparoscopic repair (LR) of rectal prolapse is potentially associated with earlier recovery and lower perioperative morbidity, as compared with open transabdominal repair (OR). Data on the long-term recurrence rate and functional outcome are limited. METHODS: Perioperative data on rectal prolapse in relation to all LRs performed between December 1991 and April 2004 were prospectively collected. The LR patients were matched by age, gender, and procedure type with OR patients who underwent surgery during the same period. Patients with previous complex abdominal surgery or a body mass index exceeding 40 were excluded from the study. Data on recurrence rate, bowel habits, continence, and satisfaction scores were collected using a telephone survey. RESULTS: A total of 111 patients (age, 56.8 +/- 18.1 years; female, 87%) underwent attempted LR. An operative complication deferred repair in two cases. Among the 111 patients, 42 had posterior mesh fixation, and 67 had sutured rectopexy (32 patients with sigmoid colectomy for constipation). Eight patients (7.2%) had conversion to laparotomy. Matching was established for 86 patients. The LR patients had a shorter hospital stay (mean, 3.9 vs 6.0 days; p < 0.0001). The 30-day reoperation and readmission rates were similar for the two groups. The rates for recurrence requiring surgery were 9.3% for LR and 4.7% for OR (p = 0.39) during a mean follow-up period of 59 months. An additional seven patients in each group reported possible recurrence by telephone. Postoperatively, 35% of the LR patients and 53% of the OR patients experienced constipation (p = 0.09). Constipation was improved in 74% of the LR patients and 54% of the OR patients, and worsened, respectively, in 3% and 17% (p = 0.037). The postoperative incontinence rates were 30% for LR and 33% for OR (p = 0.83). Continence was improved in 48% of the LR patients and 35% of the OR patients, and worsened, respectively, in 9% and 18% (p = 0.22). The mean satisfaction rates for surgery (on a scale of 0 to 10) were 7.3 for the LR patients and 8.1 for the OR patients (p = 0.17). CONCLUSIONS: The hospital stay is shorter for LR than for OR. Both functional results and recurrent full thickness rectal prolapse were similar for LR and OR during a mean follow-up period of 5 years. PMID- 16374676 TI - Board certification in pediatric cardiology. PMID- 16374675 TI - Robotic surgery training and performance: identifying objective variables for quantifying the extent of proficiency. AB - BACKGROUND: To understand the process of skill acquisition in robotic surgery and to allow useful real-time feedback to surgeons and trainees in future generations of robotic surgical systems, robotic surgical skills should be determined with objective variables. The aim of this study was to assess skill acquisition through a training protocol, and to identify variables for the quantification of proficiency. METHODS: Seven novice users of the da Vinci Surgical System engaged in 4 weeks of training that involved practicing three bimanual tasks with the system. Seven variables were determined for assessing speed of performance, bimanual coordination, and muscular activation. These values were compared before and after training. RESULTS: Significant improvements were observed through training in five variables. Bimanual coordination showed differences between the surgical tasks used, whereas muscular activation patterns showed better muscle use through training. The subjects also performed the surgical tasks considerably faster within the first two to three training sessions. CONCLUSIONS: The study objectively demonstrated that the novice users could learn to perform surgical tasks faster and with more consistency, better bimanual dexterity, and better muscular activity utilization. The variables examined showed great promise as objective indicators of proficiency and skill acquisition in robotic surgery. PMID- 16374677 TI - Catheter closure of perimembranous/membranous ventricular septal defects using the Amplatzer occluder device. AB - Perimembranous ventricular septal effects (PMVSDs) are the most common congenital cardiac defects of clinical importance. During the last decade, transcatheter closure of PMVSDs has been attempted using various occluders with variable success. This article reviews the existing literature and our experience with transcatheter closure of PMVSDs with the new Amplatzer occluder device. PMID- 16374678 TI - Hybrid pediatric cardiac surgery. AB - Minimally invasive strategies can be expanded by combining standard surgical and interventional techniques. We performed a longitudinal prospective study of all pediatric patients who have undergone hybrid cardiac surgery at the University of Chicago Children's Hospital. Hybrid cardiac surgery was defined as combined catheter-based and surgical interventions in either one setting or in a planned sequential fashion within 24 hours. Between June 2000 and June 2003, 25 patients were treated with hybrid approaches. Seventeen patients with muscular ventricular septal defects (mVSDs) (mean age, 4 months; range, 2 weeks-4 years) underwent either sequential Amplatzer device closure in the catheterization laboratory followed by surgical completion (group 1A, n = 9) or one-stage intraoperative off pump device closure (group IB, n = 8) with subsequent repair of any concomitant heart lesions. Eight patients with branch pulmonary artery (PA) stenoses (group 2) underwent intraoperative PA stenting or stent balloon dilatation along with concomitant surgical procedures. All patients survived hospitalization. Complications from the hybrid approach were mostly confined to groups 1A and 2. At a mean follow-up of 18 months, 2 group 1A patients died suddenly several months after discharge. All other patients are doing well. Hybrid pediatric cardiac surgery performed in tandem by surgeons and cardiologists is safe and effective in reducing or eliminating cardiopulmonary bypass. Patients with mVSDs who are small, have poor vascular access, or have concomitant cardiac lesions are currently treated in one setting with the perventricular approach. PMID- 16374679 TI - Wall motion velocities of abdominal aorta measured by tissue Doppler imaging in normal children. AB - Tissue Doppler imaging (TDI) offers a new technique for assessing aortic wall expansion/contraction velocities and may provide a noninvasive approach to aortic wall mechanics. The purpose of this study was to determine the normal values of abdominal aortic wall motion velocities and the effect of age on these velocities in normal children. We examined 103 normal children. Using TDI, maximum wall expansion velocity during systole (peak S) and maximum wall contraction velocity during diastole (peak D) were measured. M-mode diameter of the abdominal aorta and systolic, diastolic, and mean arterial pressures were measured. Aortic stiffness was measured as (I(n)[BP(syst)/BP(diast)])/(D(s)-D(d)/D(d), where I(n) is the natural log, D(s) is the maximal abdominal aortic diameter during systole, and D(d) is the abdominal aortic diameter at end-diastole. In all subjects, wall motion velocities of the abdominal aorta were recorded. The mean values for peak S and peak D were 4.23, 1.14 and 2.16, 0.45 cm/sec, respectively. Both peak S and peak D were low in infants and increased significantly with age (r = 0.63, p < 0.0001 and r = 0.36, p = 0.0002, respectively), systolic blood pressure (r = 0.42 and 0.47, respectively, p < 0.0001), and diastolic blood pressure (r = 0.24, p = 0.016 and r = 0.28, p = 0.0038, respectively). Aortic stiffness index of the abdominal aorta was constant with age and did not correlate with peak S or peak D. Abdominal aortic wall motion velocities can be easily assessed by TDI. Age related changes in the aortic wall motion velocities are observed in normal children. This study provides baseline information for further quantitative assessment of arterial stiffness in children with congenital or acquired heart disease. PMID- 16374680 TI - Surgical outcome of coronary artery fistulas repair in children. AB - Coronary artery fistula (CAF) is a rare congenital anomaly that can be complicated by endocarditis, myocardial infarction, or coronary aneurysms. This article reviews the clinical characteristics and surgical outcome of 10 patients with CAF in Rajae Heart Center. From 1990 to 2000, 10 patients (age, 6 months to 15 years; mean age, 8.5 years) were diagnosed with CAF by echocardiography and cardiac catheterization. Six were female and 4 were male. All patients with isolated CAF (9) were asymptomatic. One patient with associated anomaly (mitral valve prolapse with severe mitral regurgitation) had dyspnea on exertion and palpitation. Five fistulas originated from the right coronary artery-three from the left and two from the left circumflex. Drainage was to the right ventricle (7), right atrium (2), and pulmonary artery (1). The ratio of pulmonary to systemic flow ranged between 1 and 1.6. All patients had surgical ligation. In the symptomatic patient, in addition to ligation, mitral valve replacement was performed. There was no operative or late death. Follow-up evaluation (range, 1-6 years; mean, 4.2 years) showed no evidence of recurrent or residual CAF. Surgical management of CAF is a safe and effective treatment resulting in 100% survival and clousure rate. PMID- 16374681 TI - Predictors of left ventricular performance after valve replacement in children and adolescents with chronic aortic regurgitation. AB - Aortic valve replacement has been recommended in patients who have severe symptoms, in patients with extreme left ventricle (LV) dilatation (end diastolic dimension >4 SD above normal) or LV ejection fraction <50%. However, the occurrence of advanced symptoms or severe LV dilatation raises concern about irreversible LV dysfunction. This study sought to determine the influence of preoperative symptoms, LV size and function on mortality, and postoperative LV performance in children and adolescence after valve replacement for aortic regurgitation (AR). A total of 49 patients 18 years old or younger (mean, 13.9 +/ 3) who underwent valve replacement for chronic AR between 1991 and 2001 were followed up for 1-10 years (mean, 3.3 +/- 2.1). Baseline and postoperative characteristics were compared between 13 patients (group 1) with extreme LV dilatation and 34 patients (group 2) with a lesser degree of LV enlargement. Preoperative low ejection fraction (p < 0.008), extreme LV dilatation (p < 0.05), and LV end systolic dimension >4 SD above normal (p < 0.05) were predictors of persistent LV dysfunction. Extreme LV dilatation (p < 0.0003), LV end systolic dimension (p < 0.0007), and reduced LV ejection fraction (p < 0.01) predicted persistent LV dilatation. In the setting of chronic AR, preoperative symptoms, LV systolic function, and LV internal dimensions are the main predictors for persistent LV dysfunction and dilatation. Surgical correction should be performed before LV systolic dysfunction and/or extreme LV enlargement occurs. PMID- 16374682 TI - Morphogenetic differences of secundum atrial septal defects. AB - It is generally considered that the development of secundum atrial septal defect (ASDII) or oval fossa defect is the result of excessive resorption of the embryological atrial septum primum, but this does not seem to explain all defects. We investigated 58 postmortem hearts with an ASDII and 22 normal hearts from patients ranging in age from 1 day to 49 years. The different structures of the oval fossa were examined. In 86% of the specimens, the defects were the result of a malformation of the valvula foraminis ovalis or embryological atrial septum primum, and in 14% an absent superior limbus (septum secundum) was the cause of the interatrial communication. The "septum primum" ASDs were divided into four subgroups based on the degree of deficiency of the septum primum and position of the ostium secundum within the septum primum. We conclude that the morphogenesis of ASDII is variable and both septum primum and septum secundum defects occur, which may be relevant in view of genetic studies that may lead to further differentiation of patients with and without genetically determined ASDIIs. PMID- 16374683 TI - Is morbidity influenced by staging in the fontan palliation? A single center review. AB - In a retrospective study of 32 consecutive patients undergoing a total cavopulmonary connection (TCPC), we tried to determine if the trend for decreasing age at Fontan completion and reducing the intervals between the staged procedures during the past decade was associated with a change in morbidity and outcome. In 8 patients the Fontan circulation was completed in one stage and in 24 patients an intermediate step by hemi-Fontan or bidirectional cavopulmonary anastomosis was performed before Fontan completion. Mean age at TCPC and mean interval since the previous palliation have decreased significantly during the past decade. Although major complications were significantly reduced over time the occurrence and duration of postoperative pleural effusions were not. Decreasing age as well as intervals in staged Fontan palliation have beneficial influence on major complications and outcome, without significantly affecting the duration of pleural effusions at Fontan completion. PMID- 16374684 TI - Two-dimensional echocardiographic valve measurements in healthy children: gender specific differences. AB - The goal of this study was to create nomograms of echocardiographic two dimensional valve dimensions based on a large group of children without heart disease. Children aged 0-18 years underwent standard echocardiographic evaluation. Referring diagnoses were chest pain, heart murmur, or syncope. Only patients with a structurally normal heart and normal systolic and diastolic function were included. All four valves were measured at their maximal dimensions. A total of 748 children (314 girls and 434 boys) met the inclusion criteria. Mean values and standard deviations were calculated, and z value nomograms based on body surface area were developed. Surprisingly, the boys had larger valve dimensions at all ages. These valve dimension differences were statistically significant for three of four valves even after adjustment for the differences in body sizes. The difference may be due to higher circulating blood volume in boys compared to that in girls. Because the differences are subtle, they reach statistical significance only when evaluated in a large group of subjects. Presented normal value data will be helpful in following cardiology patients and evaluating intervention strategy in patients with valve hypoplasia. PMID- 16374685 TI - The effect of coenzyme Q10 on idiopathic chronic dilated cardiomyopathy in children. AB - The objective of this study was to assess the effect of coenzyme Q10 (CoQ10) as supplementation to conventional antifailure drugs on quality of life and cardiac function in children with chronic heart failure due to dilated cardiomyopathy (DCM). The study was an open-label prospective study performed in two of the largest pediatric centers in Thailand from August 2000 to June 2003. A total of 15 patients with idiopathic chronic DCM were included, with the median age of 4.4 years (range, 0.6-16.3). Presenting symptoms were congestive heart failure in 12 cases (80%), cardiogenic shock in 2 cases (13.3%), and cardiac arrhythmia in 1 case (6.7%). Sixty-one percent of patients were in the New York Heart Association functional class 2 (NYHA 2), 31% in NYHA 3, and 8% in NYHA 4. Cardiothoracic ratio from chest x-ray, left ventricular ejection fraction, and left ventricular end diastolic dimension in echocardiogram were 0.62 (range, 0.55-0.78), 30% (range, 20-40), and 5.2 cm (range, 3.8-6.5), respectively. CoQ10 was given at a dosage of 3.1 ? 0.6 mg/kg/day for 9 months as a supplementation to a fixed amount of conventional antifailure drugs throughout the study. At follow-up periods of 1, 3, 6, and 9 months, NYHA functional class was significantly improved, as was CT ratio and QRS duration at 3 and 9 months follow-up with CoQ10 when compared to the baseline and post-discontinuation of CoQ10 at 9 months (range, 4.8-10.8). However, when multiple comparisons were taken into consideration, there was no statistical significant improvement. In addition to the conventional antifailure drugs, CoQ10 may improve NYHA class and CT ratio and shorten ventricular depolarization in children with chronic idiopathic DCM. PMID- 16374686 TI - Chest radiography and the evaluation of the neonate for congenital heart disease. AB - We assessed the utility of the chest film as a screening tool for term and preterm neonates with suspected structural heart disease. Three independent observers, blinded to patient diagnosis, retrospectively evaluated the initial chest film for each of 128 consecutive neonates with suspected heart disease (gestational age, 23-42 weeks; birth weight, 500-4,621 g) who had undergone both chest radiography and echocardiography. These evaluations were subsequently compared with the results of the respective echocardiograms, read by board certified pediatric cardiologists blinded to the chest film and chest film interpretation. Kappa statistics demonstrated moderate correlation (0.42-0.48) among different observers in their interpretation of the chest films but poor correlation (0.15-0.34) between chest radiography and echocardiography. The chest film had a low sensitivity for structural heart disease (26-59%), with a negative predictive value of 46-52%. Among neonates less than 2 kg or younger than 35 weeks of gestation, the chest film had still lower sensitivity for detecting heart disease. Despite agreement among observers in chest film interpretation, the chest film does not function as a screening test for neonates with suspected heart disease, particularly in small or premature neonates. In neonates with suspected heart disease, echocardiography should be considered, even in patients with chest films that do not suggest congenital heart disease. PMID- 16374687 TI - Quantitative analysis of paradoxical interventricular septal motion following corrective surgery of tetralogy of fallot. AB - This study aimed to quantify paradoxical interventricular septal motion (PSM) among 20 patients following tetralogy of Fallot (TOF) repair without severe pulmonary regurgitation and 20 age-matched normal subjects. PSM was quantified using the echocardiography-derived paradox index. Tissue Doppler-derived strain rate was used to assess the longitudinal and radial systolic function of the interventricular septum (IVS). The tissue Doppler-derived Tei index was used to assess the global left ventricular function. Compared to the control group, the paradox index in patients after repair of TOF was significantly higher (p = 0.001), whereas the regional IVS longitudinal (p = 0.02) and radial (p = 0.001) systolic strain rate peaks were significantly reduced. The paradox index in the patient group correlated inversely with the IVS radial peak systolic strain rate (r = -0.64, p = 0.004) and positively with QRS duration (r = 0.50, p = 0.02). The left ventricular (LV) Tei-index correlated significantly with the paradox index (r = 0.71, p = 0.001) and with the septal radial systolic strain rate peak (r = 0.59, p = 0.004). We conclude that electrical delay and reduced regional septal systolic function were the main causes for paradoxical septal motion among patients following TOF repair without significant pulmonary regurgitation. The reduced LV systolic function among this group of patients is mainly secondary to diminished septal systolic function and the paradoxical septal motion. PMID- 16374688 TI - Comparison of the tissue Doppler-derived left ventricular Tei index to that obtained by pulse Doppler in patients with congenital and acquired heart disease. AB - We compared the left ventricular Tei index measured by tissue Doppler imaging (TDI) to that obtained by pulsed Doppler (PW) in patients with congenital heart disease. In 40 consecutive patients with a variety of congenital and acquired heart diseases, the left ventricular (LV) PW Doppler-derived Tei index was assessed on-line as previously described. TDI-derived anatomic curved M-mode and the unprocessed velocity trace from the basal septum were used to time the opening and closure of the mitral and aortic valves in one cardiac cycle, respectively. The TDI Tei index was calculated off-line according to the equation (isovolumetric relaxation time + isovolumetric contraction time)/ ejection time. The Tei index calculated from TDI correlated significantly with that measured by pulsed Doppler (r = 0.92, p = 0.001). The mean difference (range) between pulsed Doppler-derived Tei index and TDI-derived Tei index was 0.005 (-0.07-0.06), which was within the limits of agreements. Interobserver variability for the TDI derived Tei index was 5 +/- 3%. The TDI Tei index can be used to assess the global LV function in patients with congenital heart disease. In contrast to the PW Doppler-derived Tei index, the TDI-derived Tei index obtained from the same cardiac cycle may help to differentiate systolic from diastolic dysfunction by providing specific information on the isovolumetric intervals. PMID- 16374689 TI - Can pediatric residents interpret electrocardiograms? AB - The purpose of this study was to assess the electrocardiogram (ECG) interpretation skills of pediatric residents in a controlled environment and determine if the level of residency training (intern vs senior) improves accuracy. A list of ECG diagnoses was provided to four pediatric residency educators with instructions to categorize each diagnosis as follows: I, all residents; II, the majority of residents, including all senior residents; III, less than the majority of residents; and IV, few residents should be able to interpret correctly. Only those categories that the entire panel believed all residents (category I) or all senior residents (category II) should be able to interpret correctly were included. The test included 17 ECGs: 14 category I and 3 category II. A total of 132 residents participated: 78 interns and 54 seniors. Both groups scored below expected levels. Mean correct score among seniors was 10.9 out of the expected 17 (p < 0.001). Mean correct score for interns was 7.7 out of the expected 14 (p < 0.00l). No difference in ECG interpretation accuracy was found between residency programs. In general, pediatric residents' ECG interpretation skills are less accurate than expected. Although there is a trend toward improvement during training, senior residents fell short of the expectations of the panel. We speculate that focused education in this area will improve resident ECG interpretation and benefit patient care by (1) facilitating referral and treatment of patients with cardiovascular disease and (2) decreasing referrals for erroneous interpretations. PMID- 16374690 TI - Risk factors for interstage death after the Norwood procedure. AB - Operative survival after the Norwood procedure has significantly improved during the past 10 years. However, there remains attrition among Norwood survivors before reaching planned second-stage palliation. The purpose of this study was to evaluate potential risk factors for interstage mortality among Norwood survivors. All patients undergoing the Norwood procedure at the Medical University of South Carolina from January 1996 through January 2001 were retrospectively reviewed. Patient and procedural variables were examined as potential risk factors for interstage mortality. Among 50 Norwood survivors, 8 (16%) died prior to second stage palliation. The mean age at death was 102 +/- 72 days (median, 61; range, 35-208). By multivariate analysis, the presence of an arrhythmia in the postoperative period (p = 0.02) and decreased ventricular function at hospital discharge (p = 0.05) were identified as risk factors for interstage mortality. There remains a significant risk for interstage mortality among Norwood survivors. Patients with postoperative arrhythmias and/or decreased ventricular function at discharge are at increased risk for interstage death after Norwood procedure. More frequent follow-up and aggressive medical management of arrhythmia or decreased function may be warranted for these high-risk patients. PMID- 16374691 TI - Serum sodium levels in patients with Kawasaki disease. AB - The purpose of this study was to assess the hypothesis that lower serum sodium levels are associated with cardiovascular sequelae in patients with Kawasaki disease (KD). We used the database of the 16th nationwide survey of KD in Japan. We investigated the distribution of serum sodium levels and the relationship between serum sodium levels and cardiovascular sequelae. Of the reported cases, serum sodium levels were reported in 13,569 patients (89%). The proportion of patients with serum sodium levels 130 mEq/L or less, was greater in complete cases than in incomplete cases. The proportion of patients with serum sodium levels 130 mEq/L or less was increased with age. The largest proportion of patients with serum sodium levels 130 mEq/L or less was found in the category of 3-5 days since onset of illness. A serum sodium of level 135 mEq/L or less was an independent risk factor for cardiovascular sequelae (odds ratio, 1.79, 95% confidence interval, 1.42-2.26). Among patients with KD, there are significant differences in serum sodium levels between diagnostic categories, age, and days since the onset of illness. The sodium level may be a simple predictor of cardiovascular sequelae. PMID- 16374692 TI - Safety and efficacy of intravenous contrast imaging in pediatric echocardiography. AB - This study was performed to determine the safety and efficacy of intravenous contrast echocardiography in children attending a tertiary cardiac center. This was a prospective study to evaluate the use of Optison contrast agent in children with severely limited transthoracic echocardiographic windows. Twenty children (median age, 15 years; range, 9-18) underwent fundamental imaging (FI), harmonic imaging (HI), and HI with intravenous contrast (Optison FS-069). Endocardial border delineation was determined based on a visual qualitative scoring system (0, none: 4, excellent). Endocardial border definition was significantly improved in all patients using contrast echocardiography (FI vs Optison, p < 0.001 for each). Improved border definition was most dramatic in the apical and left ventricular (LV) free wall regions. Left ventricular ejection fraction (LVEF) was measurable in 20 patients (100%) using contrast compared to 11 (55%) with FI or HI (p < 0.05). The echocardiographic diagnosis was correctly delineated in 1 patient with a severely dyskinetic LV segment only with use of intravenous contrast and HI. No patients suffered adverse hemodynamic effects, changes in taste, or flushing episodes. Three patients experienced transient headaches. Intravenous contrast echocardiography offers an additional tool in evaluating children with very poor transthoracic echocardiographic windows. Such a strategy increases diagnostic accuracy and allows accurate LVEF determination. Adverse hemodynamic effects related to intravenous contrast are exceedingly rare. PMID- 16374693 TI - Effects of gender and aerobic fitness on cardiac autonomic responses to head-up tilt in healthy adolescents. AB - Cardiovascular autonomic responses to orthostatic challenges are affected by gender and cardiorespiratory fitness in adults. However, little is know about the effects of these factors in healthy adolescents. We studied 41 adolescents (20 boys and 21 girls) aged 12-17 years, divided into aerobic fitness tertiles based on the results of a maximal treadmill exercise test. Cardiac autonomic modulation was assessed by heart rate variability (HRV) analysis of 5-minute RR interval recordings before and after 70 degrees head-up tilt maneuver. HRV was analyzed by time (TD) and frequency domain (FD) methods. TD was analyzed by standard deviation of the RR intervals and the root mean square of successive differences of RR intervals. The power spectral components were studied at low (LF) and high (HF) frequencies and as the LF/HF ratio. We did not find any differences in TD and FD measures before and after tilt in either gender or fitness groups, except for a higher heart rate response for boys. These results suggests that cardiac autonomic responses to head-up tilt in healthy adolescents are not affected by gender or aerobic fitness. PMID- 16374694 TI - Octreotide in the management of postoperative chylothorax. AB - Chylothorax (KT) may be a complication of thoracic surgery. Its management is not well established and may comprise dietary interventions and surgery. The effectiveness of somatostatin and its analogues has been reported, although their mechanism(s) of action is unclear. We report our experience with octreotide in a series of patients with postoperative chylothorax. Eight patients with KT were treated with a continuous intravenous infusion of octreotide (OCT) at a starting dose of 0.5 microg/kg/hr. They were compared with four additional patients with KT who were treated according to the conventional approach. After a mean of 3.3 +/- 1.9 days of treatment, fluid discharge diminished dramatically. In all patients, fluid losses stopped by postoperative day 10.5 +/- 2.9 and chest tubes could be removed after 12.8 +/- 4.1 days. Compared to a small group of historical controls, OCT reduced significantly the total fluid losses (141.1 +/- 89.3 vs 396.7 +/- 151.0 ml/kg; p = 0.003) and the postoperative length of stay (p = 0.05). No patients in the group treated with OCT required parenteral nutrition (compared to all four of the controls; p = 0.002) and/or thoracenteses (compared to two of four controls). In postoperative KT, OCT seems to be at least as effective as the conventional approach. Furthermore, OCT may reduce total fluid losses and postoperative length of stay. This may have a beneficial effect on the risk of complications and on hospital costs. PMID- 16374695 TI - Phentolamine as a treatment for poor mixing in transposition of the great arteries with adequate intraatrial communication. AB - Patients with transposition of the great arteries often show poor mixing for different reasons, even after adequate balloon atrial septostomy. We present a patient with such a lesion whose clinical status improved dramatically after phentolamine was applied. We believe this improvement is due to reduction in afterload caused by the alpha(2) blocker and also possibly as a response to a presumptive effect of the drug on the diastolic function of the right ventricle, allowing more left-to-right shunt across the atrial septal defect. Both phenomena can improve cardiac output in such a situation. PMID- 16374696 TI - Portosystemic shunt with polysplenia and hypoplastic left heart syndrome. AB - We report a unique case of portosystemic shunt with polysplenia and hypoplastic left heart syndrome. The abnormal vessel connected the right renal vein and the splenic vein. The shunt flow was from the systemic vein to the portal vein before the Fontan-type operation, but it reversed and ran from portal vein to systemic vein after the Fontan-type operation. PMID- 16374697 TI - Pulmonary venous stenosis as an associated manifestation of left-sided obstructive lesions in an infant with partial atrioventricular canal defect. AB - Patients with partial atrioventricular canal defect (PAVC) who present with congestive heart failure within the first year of life are usually found to have associated left-sided obstructive lesions. We present a case of a child with PAVC who subsequently developed progressive pulmonary venous stenosis as a manifestation of her disease. PMID- 16374698 TI - Transient postnatal heart failure caused by noncompaction of the right ventricular myocardium. AB - Isolated noncompaction of the ventricular myocardium is a cardiomyopathy influencing almost exclusively the left ventricle in children and adults. We report a case of a neonate presenting with right ventricular failure caused by a noncompaction of the right ventricle. However, later course of the disease demonstrated biventricular involvement. Right ventricular noncompaction can be a cause of early postnatal right ventricular failure. PMID- 16374699 TI - A case of myocardial infarction with sumatriptan use. AB - Sumatriptan is widely used in the treatment of acute attacks of cluster headache. It is a serotonin-1 (5HT-1) agonist. Several studies have reported an association between sumatriptan use and myocardial infarction, possibly due to the generalized vasoconstrictive nature of this agent. We report a 16-year-old male patient presenting with acute inferior myocardial infarction after sumatriptan use without any known risk factors of coronary artery disease. PMID- 16374700 TI - Tetralogy of fallot with left heart hypoplasia, total anomalous pulmonary venous return, and right lung hypoplasia: role of magnetic resonance imaging. AB - We report a rare case of tetralogy of Fallot with total anomalous pulmonary venous return, left heart hypoplasia, right lung hypoplasia, and left ocular mandibular synchinesia (Marcus-Gunn phenomenon), correctly diagnosed by cardiovascular magnetic resonance imaging and successfully operated by modified Glenn anastomosis. PMID- 16374701 TI - A child case of acute right coronary obstruction due to catheter-induced coronary dissection: successful reperfusion without stenting. AB - We report on a 10-year-old child who suffered from acute right coronary obstruction due to catheter-induced coronary dissection. Immediate placement of a perfusion catheter into the obstructed right coronary artery and subsequent overnight reperfusion allowed successful recovery of the right coronary artery lumen without implantation of a stent. Follow-up angiography demonstrated spontaneous regression of the dissected coronary artery and normal right and left ventricular wall motion. The indication of stent implantation should be carefully determined in a child case of iatrogenic coronary dissection because stenting may induce coronary stenosis during growth. PMID- 16374702 TI - Amplatzer angled duct occluder for closure of patent ductus arteriosus larger than the aorta in an infant. AB - Transcatheter closure of patent ductus arteriosus (PDA) by Amplatzer duct occluder is the treatment of choice. However, closure of very large ducts in infants with low weight is a challenge for the interventionalist because a large device may obstruct the aorta or left pulmonary artery. Difficulty is also encountered in advancing the device around the curve of the right ventricular outflow tract toward the pulmonary artery; this curve is tight, more or less at a right angle in infants, leading to kinking of the sheath, which increases fluoroscopic time. This is the first reported case of a very large PDA (8.7 mm), larger than the aorta (8.2 mm), successfully closed by an Amplatzer angled duct occluder in an infant weighing 5 kg. PMID- 16374706 TI - Changes in mitotic rate and GFAP expression in the primary olfactory axis of streptozotocin-induced diabetic rats. AB - Many diabetic individuals develop anosmia but the mechanism(s) causing the dysfunction in the olfactory system is (are) unknown. Glial fibrillary acidic protein expression is reduced in diabetic retinopathy and is also reduced, with unknown consequences, in other brain regions of diabetic rats. We used immunohistochemistry and immunoblotting from untreated control and streptozotocin induced type 1 (insulin dependent) diabetic rats to investigate main olfactory epithelial mitotic rate and glial fibrillary acidic protein expression in the lamina propria of the sensory epithelium and in the olfactory bulb. Numbers of bromodeoxyuridine-positive cells were significantly lower in the diabetic sensory epithelium compared to non-diabetic controls. Immunohistochemical observations suggested a qualitative difference in glial fibrillary acidic protein expression in both regions examined especially in the olfactory bulb external plexiform layer and the lamina propria. Immunoblot analysis confirmed that the diabetic olfactory bulb and lamina propria expressed less glial fibrillary acidic protein compared to the non-diabetic control group. The lower expression levels in the olfactory bulb external plexiform layer suggested by immunohistochemistry do not reflect a change in the number of astrocytes since the numbers of S100B(+) cells were not different between the two groups. PMID- 16374707 TI - The fine-structural distribution of G-protein receptor kinase 3, beta-arrestin-2, Ca2+/calmodulin-dependent protein kinase II and phosphodiesterase PDE1C2, and a Cl(-)-cotransporter in rodent olfactory epithelia. AB - The sequentially activated molecules of olfactory signal-onset are mostly concentrated in the long, thin distal parts of olfactory epithelial receptor cell cilia. Is this also true for molecules of olfactory signal-termination and regulation? G-protein receptor kinase 3 (GRK3) supposedly aids in signal desensitization at the level of odor receptors, whereas beta-arrestin-2, Ca2+/calmodulin-dependent protein kinase II (CaMKII) and phosphodiesterase (PDE) PDE1C2 are thought to do so at the level of the adenylyl cyclase, ACIII. The Na+, K(+)-2Cl(-)-cotransporter NKCC1 regulates Cl(-)-channel activity. In an attempt to localize the subcellular sites olfactory signal-termination and -regulation we used four antibodies to GRK3, two to beta-arrestin-2, five to CaMKII (one to both the alpha and beta form, and two each specific to CaMKII alpha and beta), two to PDE1C2, and three to Cl(-)-cotransporters. Only antibodies to Cl(-) cotransporters labeled cytoplasmic compartments of, especially, supporting cells but also those of receptor cells. For all other antibodies, immunoreactivity was mostly restricted to the olfactory epithelial luminal border, confirming light microscopic studies that had shown that antibodies to GRK3, beta- arrestin-2, CaMKII, and PDE1C2 labeled this region. Labeling did indeed include receptor cell cilia but occurred in microvilli of neighboring supporting cells as well. Apical parts of microvillous cells that are distinct from supporting cells, and also of ciliated respiratory cells, immunoreacted slightly with most antibodies. When peptides were available, antibody preabsorption with an excess of peptide reduced labeling intensities. Though some of the antibodies did label apices and microvilli of vomeronasal (VNO) supporting cells, none immunoreacted with VNO sensory structures. PMID- 16374708 TI - Conditional ablation of mature olfactory sensory neurons mediated by diphtheria toxin receptor. AB - The vertebrate olfactory epithelium provides an excellent model system to study the regulatory mechanisms of neurogenesis and neuronal differentiation due to its unique ability to generate new sensory neurons throughout life. The replacement of olfactory sensory neurons is stimulated when damage occurs in the olfactory epithelium. In this study, transgenic mice, with a transgene containing human diphtheria toxin receptor under the control of the olfactory marker protein promoter (OMP-DTR), were generated in which the mature olfactory sensory neurons could be specifically ablated when exposed to diphtheria toxin. Following diphtheria toxin induced neuronal ablation, we observed increased numbers of newly generated growth associated protein 43 (GAP43)-positive immature olfactory sensory neurons. OMP-positive neurons were continuously produced from the newly generated GAP43-positive cells. The expression of the signal transduction components adenylyl cyclase type III and the G-protein alpha subunit G(alpha olf) was sensitive to diphtheria toxin exposure and their levels decreased dramatically preceding the disappearance of the OMP-positive sensory neurons. These data validate the hypothesis that OMP-DTR mice can be used as a tool to ablate the mature olfactory sensory neurons in a controlled fashion and to study the regulatory mechanisms of the neuronal replacement. PMID- 16374709 TI - Morphological and molecular features of the mammalian olfactory sensory neuron axons: What makes these axons so special? AB - The main organization and gross morphology of the mammalian olfactory primary pathway, from the olfactory epithelium to the olfactory bulb, has been initially characterized using classical anatomical and ultrastructural approaches. During the last fifteen years, essentially thanks to the cloning of the odorant receptor genes, and to the characterization of a number of molecules expressed by the olfactory sensory neuron axons and their environment, significant new insights have been gained into the understanding of the development and adult functioning of this system. In the course of these genetic, biochemical and neuroanatomical studies, however, several molecular and structural features were uncovered that appear somehow to be unique to these axons. For example, these axons express odorant receptors in their terminal segment, and transport several mRNA species and at least two transcription factors. In the present paper, we review these unusual structural and molecular features and speculate about their possible functions in the development and maintenance of the olfactory system. PMID- 16374710 TI - Morphological and functional plasticity of olfactory ensheathing cells. AB - In the primary olfactory pathway, olfactory ensheathing cells (OECs) extend processes to envelop bundles of olfactory axons as they course towards their termination in the olfactory bulb. The expression of growth-promoting adhesion and extracellular matrix molecules by OECs, and their spatially close association with olfactory axons are consistent with OECs being involved in promoting and guiding olfactory axon growth. Because of this, OECs have been employed as a possible tool for inducing axonal regeneration in the injured adult CNS, resulting in significant functional recovery in some animal models and promising outcomes from early clinical applications. However, fundamental aspects of OEC biology remain unclear. This brief review discusses some of the experimental data that have resulted in conflicting views with regard to the identity of OECs. We present here recent findings which support the notion of OECs as a single but malleable phenotype which demonstrate extensive morphological and functional plasticity depending on the environmental stimuli. The review includes a discussion of the normal functional role of OECs in the developing primary olfactory pathway as well as their interaction with regenerating axons and reactive astrocytes in the novel environment of the injured CNS. The use of OECs to induce repair in the injured nervous system reflects the functional plasticity of these cells. Finally, we will explore the possibility that recent microarray data could point to OECs assuming an innate immune function or playing a role in modulating neuroinflammation. PMID- 16374711 TI - The alpha6 integrin subunit in the developing mouse olfactory bulb. AB - Integrins are heterodimeric cell surface receptors that mediate developmental events by binding extracellular matrix ligands. Several lines of evidence suggest a role for integrins, specifically the alpha 6 subunit, in neuronal migration, neurite outgrowth, and axon guidance during olfactory development. Therefore, we undertook an analysis of the expression of the alpha 6 subunit in the olfactory system of the embryonic and early postnatal mouse to understand the role it may play during neural development. In addition, as a functional assay we examined the developmental effects of the loss of this subunit on olfactory development by analyzing an alpha 6 knockout (alpha 6-/-). Immunohistochemical analyses and confocal microscopy were used to examine alpha 6 expression in the CD-1 embryonic and early postnatal olfactory system and also to examine the organization of the olfactory system in the alpha 6-/- mouse. In CD-1 mice from E13 to E17, alpha 6 localizes in radial patterns extending from the core of the olfactory bulb to the nerve layer and colocalizes with RC2, an antibody specific for radial glia. By the day of birth (P0; approximately E19), expression is limited to the external plexiform layer and the olfactory nerve layer, where it colocalizes with laminin and p75. In the alpha 6-/- mouse, areas of ectopic granule cells were observed in the mitral cell layer of the olfactory bulb. These ectopias coincided with areas of disorganization of the radial glial processes and breaks in the mitral cell layer. These observations suggest a role for alpha 6 integrin in neural migration during olfactory development, likely secondary to organization of the radial glial scaffold. PMID- 16374712 TI - Morituri te salutant? Olfactory signal transduction and the role of phosphoinositides. AB - During the past 150 years, researchers have investigated the cellular, physiological, and molecular mechanisms underlying the sense of smell. Based on these efforts, a conclusive model of olfactory signal transduction in the vertebrate's nose is now available, spanning from G-protein-mediated odorant receptors to ion channels, which are linked by a cyclic adenosine 3',5' monophosphate-mediated signal transduction cascade. Here we review some historical milestones in the chronology of olfactory research, particularly emphasising the role of cyclic nucleotides and inositol trisphosphate as alternative second messengers in olfactory cells. We will describe the functional anatomy of the nose, outline the cellular composition of the olfactory epithelium, and describe the discovery of the molecular backbone of the olfactory signal transduction cascade. We then summarize our current model, in which cyclic adenosine monophosphate is the sole excitatory second messenger in olfactory sensory neurons. Finally, a possible significance of microvillous olfactory epithelial cells and inositol trisphosphate in olfaction will be discussed. PMID- 16374714 TI - Ultrastructural localization of alpha-galactose-containing glycoconjugates in the rat vomeronasal organ. AB - Binding sites of Griffonia simplicifolia I-B4 isolectin (GS-I-B4), which recognizes terminal alpha-galactose residues of glycoconjugates, were examined in the juxtaluminal region of the rat vomeronasal sensory epithelium and its associated glands of the vomeronasal organ, using a lectin cytochemical technique. Lowicryl K4M-embedded ultra-thin sections, which were treated successively with biotinylated GS-I-B4 and streptavidin-conjugated 10 nm colloidal gold particles, were observed under a transmission electron microscope. Colloidal gold particles, which reflect the presence of terminal alpha-galactose containing glycoconjugates, were present in vomeronasal receptor neurons in the sensory epithelium and secretory granules of acinar cells of associated glands of the epithelium. Quantitative analysis demonstrated that the density of colloidal gold particles associated with sensory cell microvilli that projected from dendritic endings of vomeronasal neurons was considerably higher than that of microvilli that projected from neighboring sustentacular cells. The same was true for the apical cytoplasms of these cells just below the microvilli. These results suggest that of the sensory microvilli and dendritic endings contained a much larger amount of the alpha-galactose-containing glycoconjugates, compared with those in sustentacular microvilli. Further, biochemical analyses demonstrated several vomeronasal organ-specific glycoproteins with terminal alpha-galactose. PMID- 16374713 TI - Solitary chemoreceptor cell proliferation in adult nasal epithelium. AB - Nasal trigeminal chemosensitivity in mice and rats is mediated in part by solitary chemoreceptor cells (SCCs) in the nasal epithelium (Finger et al., 2003). Many nasal SCCs express the G-protein alpha-gustducin as well as other elements of the bitter-taste signaling cascade including phospholipase Cbeta2, TRPM5 and T2R bitter-taste receptors. While some populations of sensory cells are replaced throughout life (taste and olfaction), others are not (hair cells and carotid body chemoreceptors). These experiments were designed to test whether new SCCs are generated within the epithelium of adult mice. Wild type C57/B6 mice were injected with the thymidine analog 5-bromo-2'-deoxyuridine (BrdU) to label dividing cells. At various times after injection (1-40 days), the mice were perfused with 4% paraformaldehyde and prepared for dual-label immunocytochemistry. Double labeled cells were detected as early as 3 days post BrdU injection and remained for as long as 12 days post-injection suggesting that SCCs do undergo turnover like the surrounding nasal epithelium. No BrdU labeled cells were detected after 24 days suggesting relatively rapid replacement of the SCCs. PMID- 16374715 TI - The vomeronasal organ of greater bushbabies (Otolemur spp.): species, sex, and age differences. AB - The present study examined interspecies, intersexual, and age-related changes in size of the vomeronasal neuroepithelium (VNNE) of two species of greater bushbabies (genus Otolemur, Infraorder Lorisiformes, Suborder Strepsirrhini). Tissue blocks containing the vomeronasal organs of nine O. crassicaudatus (8 adults, 1 neonate) and ten O. garnettii (9 adults, 1 neonate) were studied by means of serial paraffin sectioning and computer-based reconstruction of VNNE volume. In addition, the immunoreactivity of the VNNE to two neuronal markers, neuron-specific beta tubulin (BT) and olfactory marker protein (OMP) was compared between species, sexes, and ages. Results indicated that a clear VNNE is present at birth in both species, and OMP immunoreactivity was verified in O. garnettii at birth. Male and female adults of both species showed OMP-immunoreactive and BT immunoreactive neurons in the VNNE. Immunohistochemical findings indicated that all males and the youngest females had the thickest VNNE, especially at the marginal junctions with the receptor-free epithelium. Results of a 2-way Analysis of Variance (ANOVA, species x sex) revealed no significant differences in VNNE length or volume between species, but O. crassicaudatus had significantly (p < 0.05) greater palatal length. Significant (p < 0.05) differences also were found between sexes in VNNE volume, but no significant differences in palatal length or VNNE length. The distribution of VNNE volume against age indicated that the sex differences were more pronounced in O. crassicaudatus than O. garnettii. For both species and sexes, distribution of VNNE volume against age suggested an age related reduction in volume. These findings demonstrate postnatal plasticity in VNNE size in Otolemur that is reminiscent of that found for olfactory structures in some rodents. Bushbabies or other strepsirrhine primates may offer an opportunity for further understanding of behavioral correlates of VNNE postnatal plasticity, which may represent primitive functional characteristics of the order Primates. PMID- 16374716 TI - The expression pattern of four odorant-binding proteins in male and female silk moths, Bombyx mori. AB - Four recombinant odorant-binding proteins of Bombyx mori, pheromone-binding protein (PBP), general odorant-binding protein 1 (GOBP1), general odorant-binding protein 2 (GOBP2) and antennal binding protein X (ABPX), were expressed in E. coli and used to raise polyclonal antisera. Immunoblots of antennal homogenates showed that these antisera were specific. In Western blot analysis and immunocytochemical labelling experiments, the sera against recombinant PBP and GOBP2 of B. mori gave identical results as sera against native PBP and GOBP2 of Antheraea polyphemus, respectively, thus confirming earlier results obtained with the latter. Labelling consecutive cross sections of various sensillum types with all four antisera revealed different labelling patterns in male and female sensilla (s.) trichodea and s. basiconica. Long s. trichodea in males and females represented uniform labelling types, whereas for short s. trichodea, s. intermedia, and s. basiconica a great variety of labelling patterns was observed, some being more common than others. Long s. trichodea, which in males are uniformly tuned to the pheromone components bombykol and bombykal, all strongly expressed PBP; labelling with antisera against the other three odorant-binding proteins hardly was above background, only in some hairs GOBP1 was expressed somewhat more strongly. Long s. trichodea of females, which respond specifically to linalool and benzoic acid, showed a different labelling pattern. Here, we observed strong labelling with antibodies against GOBP2 and medium labelling with anti-GOBP1, sometimes with anti-ABPX. S. basiconica in both sexes most commonly co-expressed GOBP1 and GOBP2, but other patterns were occasionally found, with some of them showing PBP expression, also in females. The great variety of labelling types in short s. trichodea, s. intermedia, and s. basiconica suggests a similar variety of functional subtypes as observed in plant odour-sensitive sensilla of other moth species. PMID- 16374718 TI - Intercellular trafficking of the major tegument protein VP22 of bovine herpesvirus-1 and its application to improve a DNA vaccine. AB - Intercellular spread of bovine herpesvirus-1 (BHV-1) VP22 was demonstrated in living COS-7 cells transfected with a plasmid expressing VP22-YFP (yellow fluorescence protein) and CFP (cyan fluorescence protein) bicistronically. The intercellular trafficking property of VP22 was localized to the C-terminal portion of the molecule (amino acids 121-258; VP22-C). Plasmids encoding a truncated form of BHV-1 glycoprotein D (tgD) fused to VP22, VP22-C, or the N terminal portion of VP22 (amino acids 1-120; VP22-N) were constructed. Mice immunized with plasmid encoding tgD-VP22 or tgD-VP22-C developed stronger immune responses when compared to animals immunized with plasmid encoding tgD or tgD fused to tgD-VP22-N. PMID- 16374719 TI - Complete nucleotide sequence of the RNA2 of the crinivirus tomato chlorosis virus. AB - The complete sequence of genomic RNA2 of Tomato chlorosis virus (ToCV; genus Crinivirus, family Closteroviridae), isolate AT80/99 from Spain, was determined and compared with those from the other members of the genus sequenced to date. RNA2 is 8244 nucleotides (nt) long and putatively encodes nine ORFs that encompass the hallmark gene array of the family Closteroviridae, which includes a heat shock protein 70 family homologue, a 59 kDa protein, the coat protein, and a diverged coat protein. Phylogenetic analysis confirmed assignment of ToCV in the genus Crinivirus, being most similar to sweet potato chlorotic stunt virus and cucurbit yellow stunting disorder virus. PMID- 16374720 TI - AACR-NCI-EORTC--17th symposium: Molecular targets and cancer therapeutics. 14 18th November 2005, Philadelphia, PA, USA. PMID- 16374721 TI - AACR-NCI-EORTC--17th symposium: Molecular targets and cancer therapeutics. Imaging agents, drug delivery and targeting, and drug updates. 14-18th November 2005, Philadelphia, PA, USA. PMID- 16374722 TI - AACR-NCI-EORTC--17th symposium: Molecular targets and cancer therapeutics. Emerging targets and new agents from academia. 14-18th November 2005, Philadelphia, PA, USA. PMID- 16374723 TI - AACR-NCI-EORTC--17th symposium: Molecular targets and cancer therapeutics. Drug highlights. 14-18th November 2005, Philadelphia, PA, USA. PMID- 16374724 TI - American Heart Association--scientific sessions 2005. 13-16 November 2005, Dallas, TX, USA. PMID- 16374725 TI - American Heart Association--scientific sessions 2005. Potential new drugs for various indications. 13-16 November 2005, Dallas, TX, USA. PMID- 16374726 TI - Neurology--18th world congress. 5-11 November 2005, Sydney, Australia. PMID- 16374727 TI - Asian Pacific Society of Respirology--tenth congress and first joint congress with the American College of Chest Physicians. 11-14 November 2005, Guangzhou, China. PMID- 16374728 TI - Peptides and therapeutic proteins--SRI conference. New opportunities for macromolecule development, delivery and commercialization. 17-18 November 2005, Philadelphia, PA, USA. PMID- 16374729 TI - Metabolic profiling--IBC's inaugural meeting. Using metabolomics to accelerate drug discovery and development. 14-15 November 2005, Durham, NC, USA. PMID- 16374730 TI - BioBusiness Summit 2005. Global clinical trials in India--prospects and challenges. 14-15 November 2005, New Delhi, India. PMID- 16374731 TI - The Wnt signaling pathway as a target for the treatment of neurodegenerative disorders. AB - Although significant progress in understanding brain function has been made in the last 15 years, the unmet medical need for effective therapeutic treatment of devastating neurodegenerative disorders is still enormous and represents a formidable challenge at the beginning of the 21st century. With the recent accumulation of evidence that the Wnt signaling pathway might be impaired in such diseases, a new avenue for potential therapeutic intervention has been opened which comprises many putative drug targets. PMID- 16374732 TI - The discovery of finger loop inhibitors of the hepatitis C virus NS5B polymerase: status and prospects for novel HCV therapeutics. AB - Millions of people worldwide are infected with the hepatitis C virus (HCV), for which the main current therapy is associated with severe side effects, limited efficacy, is contraindicated in many patients, and has issues of compliance. As a result, new HCV therapies that are more effective and better tolerated are required. This review describes the discovery of a new class of allosteric, non nucleoside HCV polymerase inhibitors that demonstrate potential for the development of novel anti-HCV therapeutics. PMID- 16374733 TI - Improving drug manufacturing with process analytical technology. AB - Within the process analytical technology (PAT) framework, as presented in the US Food and Drug Administration guidelines, the aim is to design, develop and operate processes consistently to ensure a pre-defined level of quality at the end of the manufacturing process. Three PAT implementation scenarios can be envisaged. Firstly, PAT could be used in its most modest version (in an almost non-PAT manner) to simply replace an existing quality control protocol (eg, using near-infrared spectroscopy for an in-process quality control, such as moisture content). Secondly, the use of in-process monitoring and process analysis could be integrated to enhance process understanding and operation for an existing industrial process. Thirdly, PAT could be used extensively and exclusively throughout development, scale-up and full-scale production of a new product and process. Although the first type of implementations are well known, reports of the second and third types remain scarce. Herein, results obtained from PAT implementations of the second and third types are described for two industrial processes for preparing bulk active pharmaceutical ingredients, demonstrating the benefits in terms of increased process understanding and process control. PMID- 16374734 TI - Abiraterone. Cougar Biotechnology. AB - Abiraterone is an oral, selective, steroidal inhibitor of cytochrome P450(17alpha) being developed by Cougar Biotechnology Inc for the potential treatment of prostate cancer. Phase I clinical trials of abiraterone have been conducted and the design of phase I/II clinical trials is being finalized. PMID- 16374736 TI - [Pancreatic elastase-1 in the stool for the determination of exocrine pancreas function of postoperative patients with early enteral nutrition -- a pilot study]. AB - BACKGROUND AND OBJECTIVE: There are only incomplete data about exocrine pancreatic function after major gastrointestinal surgery. Early enteral nutrition with high-molecular diets is increasingly administered to these patients. A knowledge of exocrine secretion of the pancreas is crucial, because pancreatic enzymes are needed for the adsorption of the high-molecular diets. To determine the exocrine pancreatic secretion the fecal elastase-1 is a sensitive method for measuring exocrine pancreatic secretion. PATIENTS AND METHODS: In a prospective study fecal elastase-1 was measured in patients on enteral nutrition (n=12) early after major gastrointestinal operations. They were given a high molecular diet via an intraoperatively placed small-needle catheter jejunostomy, starting 12 hours after operation with a continuous infusion of 20 ml/h (1 kcal/ml),increased to 80 ml/h during the next few days. Samples from the first and second stools after beginning the enteral nutrition were taken for measuring the fecal elastase 1. RESULTS: All patients fed enterally had no relevant feeding-associated complications and no diarrhea. The elastase-1 concentrations were normal in both the first and the second stool samples in all patients (normal stool elastase is > 200 g/g). The average elastase concentration in the first stool sample was 361,4 mg/g (median: 317 mg/g) and 454 mg/g in the second (median: 466,6 mg/g). Thus no exocrine pancreatic insufficiency was detected in any of the patients. CONCLUSION: The results of elastase-1 in stool demonstrate that severe exocrine pancreas dysfunction is not generally present in patients on early enteral nutrition after major gastrointestinal surgery. PMID- 16374737 TI - [Haemorrhagic diathesis as an early symptom of systemic amyloidosis]. AB - HISTORY: A 63-year-old man, known to have Bechterev's disease was admitted because of weight gain and nocturnal sweating. He also had signs of heart failure with progressive exertional dyspnoea. Many months previously numerous bleedings had occurred in the skin, predominantly the face (periorbital and perioral), the inguinal region and the penis. INVESTIGATIONS: Extensive diagnostic test failed to find any evidence of tumor. The Rumpel-Leede tourniquet test was positive, while platelet functions was normal, suggesting vascular disease. The skin biopsy showed many perivascular amyloid deposits (AL). Immunochemical differentiation also demonstrated the same amyloid in rectal and hepatic biopsies. But there was AA amyloid in a hepatic artery. Immunophoresis indicated a biclonal gammopahty of unknown significance. DIAGNOSIS, TREATMENT AND COURSE: These findings indicated the diagnosis of primary AL amyloidosis associated with a gammopathy of unknown significance and a secondary AA amyloidosis in the presence of chronic Bechterev s disease. The clinical picture also showed cardiac complications, predominantly heart failure and numerous previous myocardial infarctions without S-T elevations. Echocardiography, which revealed marked thickening of the left ventricle with a restrictive filling pattern, suggested cardiac co-morbidity. The patient underwent chemotherapy with melphalan and prednisone but had a sudden cardiac death. CONCLUSION: In a case of bleeding of unknown cause systemic amyloidosis should be considered in the differential diagnosis. A tendency towards bleeding, as in this patient, may be the first sign of amyloidosis, which ist often diagnosed quite late in the course of the disease. PMID- 16374738 TI - [A 46-year-old-patient with granulomatous prostatitis, arthralgia and haemorrhagic rhinitis]. AB - HISTORY AND ADMISSION FINDINGS: A 46-year-old, having developed developed gross haematuria and pain in the genital region, was referred to a urological unit. Because palpation of his prostatic gland was abnormal, a transurethral prostatic resection was performed. The histology showed a necrotizing and granulomatous inflammation of the prostate with small-vessel vasculitis and eosinophilic infiltrates. As the patient had also developed arthralgia and symptoms in the upper respiratory tract, he was admitted to our hospital. The test for c-ANCA with specifity for proteinase 3 was positive. There was no history or symptoms of asthma. DIAGNOSIS: Wegener's granulomatosis. TREATMENT AND COURSE: To achieve remission bolus therapy with cyclophosphamide and prednisolone was initiated and planned to be the first of three pulses But when the patient developed epididymitis and a neuropathy associated with Wegener"s granulomatosis, the dose was increased and cyclophosphamide was given for a longer period. CONCLUSION: The early manifestation of Wegener"s granulomatosis at the prostatic gland is rare in this disease. This case presented with an impressive variation of symptoms, which may occur especially in the early course of vasculitis. The findings of c-ANCA with specifity for proteinase 3 is important for the diagnosis of this disease. PMID- 16374739 TI - [Significance of the identification of hereditary cancer for routine oncological diagnosis]. AB - Genetic factors are likely to be the primary determinants of the cancer seen in around 5 - 10 % of all cancer patients. More than 20 inherited cancer syndromes are now known. For many of these cases molecular genetic diagnostic may be accomplished. Within the scope of genetic counselling and testing, which can validate the genetic background of cancer, the interdisciplinary cooperation of Primary Care Physicians (PCP) and human geneticists is necessary. The results of our empirical study on risk identification in familial cancer suggest that PCP in the acute care inform cancer patients only to a little extent about the possible genetic background of their disease. PCP also very rarely initiate genetic counselling and testing, which would be desirable for preventive reasons. PMID- 16374741 TI - [Monoclonal antibodies improve therapy of relapsing multiple sclerosis. Molecular basis and clinical results of anti-VLA4 (natalizumab) therapy]. PMID- 16374742 TI - [Depression, a widespread disease. Epidemiology, care situation, diagnosis, therapy and prevention]. AB - According to a WHO study depressive disorders are the worldwide leading cause for life years lived with disability. Beside the direct costs of care, enormous indirect costs are caused by disablement and permanent incapacity to work. Furthermore, depressive disorders are often accompanied by suicidal behaviour, and they increase the mortality rate due to cardiovascular disorders. Despite their outstanding importance, there is a huge need to optimize diagnostic and therapeutic procedures in primary care, because depression is often masked by somatic symptoms. To achieve a correct diagnosis, the medical practitioner must actively explore the core criteria of depressed mood, diminished interest and pleasure, as well as fatigue and loss of energy. Improvement of early recognition and treatment of depressive patients is thus the main objective of modern awareness programs such as the "German Alliance against Depression", which was established in the framework of the German Research Network on Depression and Suicidality. PMID- 16374743 TI - [Clarification of patients at risk from the use of laminated stents]. PMID- 16374745 TI - [Significance of kidney biopsies for nephrology. Re: the article from DMW 36/2005]. PMID- 16374746 TI - [The medical-industrial complex: ethical implications. Re: the article from DMW 30/2005]. PMID- 16374748 TI - From obstetric ultrasound to ultrasonographic obstetrics. PMID- 16374749 TI - Feasibility and reproducibility of an image-scoring method for quality control of fetal biometry in the second trimester. AB - OBJECTIVES: The need for training programs and certification processes in fetal ultrasound has become obvious. The purpose of this study was to evaluate the feasibility of a score-based quality control system for fetal biometry in the second trimester. METHODS: Standard measurements of biparietal diameter and head circumference, abdominal circumference, and femur length at 20-24 weeks had been made by four operators using the same ultrasound machine. Twenty-five of each of the cephalic, abdominal and femoral images with the calipers in place were selected arbitrarily from each operator's ultrasound database and anonymized. These 300 images were analyzed by three experienced reviewers blinded to the operator's identity. Each image was first evaluated subjectively and then scored according to six criteria for abdominal and cephalic measurements and four criteria for femur length making a six-point score for abdominal and cephalic biometry and a four-point score for femur length. For subjective evaluation, inter-reviewer differences were analyzed using percentage agreement and adjusted kappa. For objective evaluation, a difference in scoring of one point or less among reviewers was considered good agreement. Intrareviewer variability was assessed using 40 images of each type of examination selected arbitrarily. RESULTS: The distribution of scores was similar between reviewers. One operator obtained significantly lower scores whereas the other three had good and comparable results. There was no statistical difference in the mean score attributed by each reviewer and agreement was good in 84-90% of the cases. Intrareviewer agreement was good in 90-100% of the cases, with similar scores for each reviewer. CONCLUSION: A quality control policy based on image scoring is feasible and allows for fair to good inter- and intrareviewer reproducibility. The potential contribution of this approach to assess the quality of routine ultrasound examinations should be tested on a larger scale. PMID- 16374750 TI - Ultrasound and color power Doppler in the detection of metastatic omentum: a prospective study. AB - OBJECTIVES: To analyze prospectively the ability of ultrasound to detect metastatic omentum in patients with suspicious pelvic masses and to describe the sonographic features of metastatic omental disease. METHODS: One hundred and eighty-four patients were evaluated preoperatively by ultrasound examination and of these 173 were used in the analysis. We defined as an ultrasound-positive examination one which visualized intra-abdominal aperistaltic solid tissue, located above the bowel loops and below the anterior peritoneal surface. Preoperative sonographic findings were compared with pathological results. RESULTS: Sonographic detection of metastatic omentum was achieved in 104 of 173 patients (60.1%), appearing as either solid aperistaltic tissue (80.8% of cases), or as solid discrete nodules (19.2%). When considering the echostructure of the surrounding bowel loops, this tissue appeared hypoechoic in 46 (44.2%) cases and isoechoic or slightly hyperechoic in the other 58 (55.8%) cases. In the overall series, the negative and positive predictive values (NPV and PPV) and the accuracy of ultrasound examination were 92.7%, 91.3% and 91.9%, respectively. When considering only the group of ovarian tumors, the NPV, PPV and accuracy were 91.9%, 94.6%, and 93.8%, respectively. CONCLUSION: Ultrasound examination is highly accurate in detecting metastatic omental involvement in cases with suspicious pelvic masses. PMID- 16374751 TI - Fetal magnetic resonance imaging in the prenatal diagnosis of cerebellar hemorrhage. AB - We report the case of a fetus with a sonographic mid-gestation diagnosis of hyperechogenic cerebellum suspected to be of hemorrhagic origin on fetal brain magnetic resonance imaging (MRI). No etiological factors for fetal hemorrhage were found other than a maternal heterozygocity for factor V Leiden. Following termination of the pregnancy, autopsy confirmed the prenatal diagnosis of massive cerebellar hemorrhage without underlying vascular anomaly. As an additional tool to ultrasonography, fetal brain MRI can affirm the hemorrhagic origin of hyperechogenic cerebellar lesions, especially by showing a high signal on T1 weighted images. PMID- 16374752 TI - Suboptimal Down syndrome screening test interpretation. PMID- 16374753 TI - First-trimester maternal serum level of pregnancy-associated plasma protein-A is an independent predictor of fetal maxillary bone length. AB - OBJECTIVE: To derive a nomogram of fetal maxillary bone length (MAX) for a Chinese population and to study whether first-trimester maternal serum levels of pregnancy-associated plasma protein-A (PAPP-A) is an independent predictor of fetal MAX. METHODS: This was a prospective observational study over 10 months examining Chinese women with a singleton pregnancy who attended the first trimester screening program for Down syndrome. The subjects had the fetal crown rump length (CRL), nuchal translucency, MAX, maternal serum PAPP-A and free beta human chorionic gonadotropin (fbeta-hCG) levels measured. A nomogram of MAX was derived using normal pregnancies with reliable dates. The correlations between MAX, CRL, PAPP-A and fbeta-hCG levels were studied, after correction for gestational dependency using Z-score transformation for the ultrasound markers (Z MAX and Z-CRL), and logarithmic transformation of multiple of gestation-specific medians for the biochemical markers (log10PAPP-A multiples of the median (MoM) and log10fbeta-hCG MoM), using the Pearson test and multiple regression analysis. RESULTS: During the study period, 607 Chinese women met the inclusion criteria. The mean gestational age at the first-trimester screening was 12 + 4 weeks (SD, 4 days). Curve estimation analysis showed that a linear relationship fit best between MAX and CRL (MAX (mm) = -0.01 + 0.101 * CRL (mm); r = 0.826; P < 0.0001), and between MAX and gestational age (MAX (mm) = -8.465 + 0.170 * gestational age (day); r = 0.754; SD = 0.71; P < 0.0001). There were significant positive correlations between Z-MAX and Z-CRL (r = 0.627; P = < 0.0001), Z-MAX and log10PAPP-A MoM (r = 0.239; P = < 0.0001). Multiple regression analysis showed that both Z-CRL (P = < 0.0001) and log10 PAPP-A MoM (P = 0.048) were independent predictors for Z-MAX. CONCLUSION: Both CRL and maternal levels of PAPP-A are independent predictors of the fetal MAX measured during the first trimester. The positive correlation between PAPP-A levels and fetal MAX is consistent with the known biological function of PAPP-A on bone growth. This association must be adjusted for when both PAPP-A and MAX are combined for screening of Down syndrome. PMID- 16374754 TI - Assessment of ovarian reserve using the inversion mode. PMID- 16374755 TI - A novel approach to first-trimester screening for early pre-eclampsia combining serum PP-13 and Doppler ultrasound. AB - OBJECTIVE: To investigate the value of maternal serum placental protein 13 (PP 13) measurement and uterine artery Doppler during first-trimester screening in the prediction of early pre-eclampsia. METHODS: This was a nested case-control prospective study of pregnancies at 11 + 0 to 13 + 6 weeks of gestation. The pulsatility index (PI) of blood flow in the uterine arteries and the maternal serum concentration of PP-13 were measured in 10 women who went on to develop pre eclampsia that necessitated delivery before 34 weeks, and in 423 unaffected women. Results were expressed as multiples of the gestation-specific median in controls (MoM). A logistic regression model was used to predict detection and false-positive rates. RESULTS: In the cases that developed pre-eclampsia requiring delivery before 34 weeks, compared with the unaffected pregnancies, the median uterine artery PI was higher (1.43 MoM) and the median serum PP-13 level was lower (0.07 MoM; P < 0.001, Wilcoxon rank sum test for both). Modeling predicted that for a 90% detection rate of pre-eclampsia requiring delivery before 34 weeks, the false-positive rate of screening by PP-13 was 12%, by uterine artery PI was 31% and by a combination of the two methods was 9%. A policy of contingency screening, whereby all women are screened by maternal serum PP-13 and only the 14% at highest risk are then screened by Doppler, achieved a detection rate of 90% with an overall false-positive rate of 6%. CONCLUSION: Effective screening for pre-eclampsia requiring delivery before 34 weeks can potentially be provided by assessment of a combination of maternal serum PP-13 and uterine artery Doppler in the first trimester of pregnancy. PMID- 16374756 TI - Prenatal prediction of survival in isolated left-sided diaphragmatic hernia. AB - OBJECTIVE: To investigate the potential value of antenatally determined intrathoracic herniation of the liver and the ratio of fetal lung area to head circumference (LHR) in the prediction of postnatal survival in isolated, left sided congenital diaphragmatic hernia (CDH). METHODS: In this multicenter study, we searched the antenatal-CDH-Registry for fetuses with isolated left-sided CDH that were liveborn after 30 weeks of gestation and were followed up postnatally for at least 3 months after discharge from hospital. The patients were subdivided into those with and those without intrathoracic herniation of the liver and into subgroups according to the LHR that had been measured between 22 + 0 and 28 + 0 weeks of gestation. The rate of survival in each group and subgroup of patients was determined and compared. RESULTS: The registry search identified 184 patients that fulfilled the entry criteria. Logistic regression analysis demonstrated that significant predictors of survival were the presence or absence of intrathoracic herniation of the liver and the LHR. In 86 cases there was liver herniation (Group A) and in 98 the liver was confined to the abdomen (Group B). In Group A, the survival rate was 50.0% and was significantly associated with LHR; in Group B the survival rate was 76.5% and was unrelated to LHR. CONCLUSION: In isolated, left-sided CDH, the postnatal mortality rate is substantially higher if there is intrathoracic herniation of the liver. In fetuses with liver herniation, measurement of LHR at 22-28 weeks of gestation is useful in the prediction of subsequent survival. PMID- 16374757 TI - Cardiac screening examination of the fetus: guidelines for performing the 'basic' and 'extended basic' cardiac scan. PMID- 16374758 TI - The non-surgical management of ectopic pregnancy. AB - Ectopic pregnancy can now be detected at earlier gestations in asymptomatic women. As a consequence conservative treatment strategies may be more appropriate than surgical intervention. This review aims to discuss the diagnosis and the non surgical management options for ectopic pregnancy, in particular expectant management and the use of methotrexate. PMID- 16374759 TI - Acoustic streaming in two endometriomas: the exception to the rule? AB - The presence of acoustic streaming in ovarian cysts can be determined by ultrasound imaging and provides an indication of the viscosity of the cyst content. It has previously been demonstrated that the absence of acoustic streaming in endometriomas could be a pathognomonic feature of endometriomas. In this report we demonstrate the presence of acoustic streaming in two endometriomas. PMID- 16374761 TI - Re: in-utero intervention for hypoplastic left heart syndrome: for which fetus and for what? PMID- 16374763 TI - Transvaginal ultrasound after first-trimester uterine evacuation reduces the incidence of retained products of conception. AB - OBJECTIVE: To assess the incidence of retained products of conception (RPOC) in relation to transvaginal ultrasound performed after first-trimester uterine evacuation. METHODS: This was a prospective randomized study involving 809 women undergoing first-trimester uterine evacuation. The study group included 404 women in whom transvaginal sonography was performed at the end of the surgical procedure and the control group contained 405 women who did not undergo ultrasound examination. Initially, in the study group, recurettage was immediately performed if the endometrium appeared irregular but latterly only if endometrial thickness was > or = 8 mm. The patients were followed up by gynecological and ultrasound examinations 5-8 days following the surgical procedure. RESULTS: The total complication rate was 4.3%. RPOC presented in three women in the study group (0.7%) and in 15 women in the control group (3.7%, P < 0.05). Vaginal bleeding requiring hospitalization occurred in two women in the study group (0.5%) vs. seven in the control group (1.7%, P = 0.2). Endometritis was diagnosed in one woman in the study group (0.2%) vs. six in the control group (1.5%) and uterine perforation occurred in one woman in the control group vs. none in the study group. There were no cases of RPOC in women who had an endometrial thickness of < 8 mm as demonstrated by ultrasound at the end of the surgical procedure. CONCLUSION: Transvaginal sonography immediately following first-trimester uterine evacuation may reduce the incidence of RPOC and the total complication rate. When the endometrial thickness is > or = 8 mm at the end of suction curettage, an attempt at re-evacuation of the uterine cavity is indicated. PMID- 16374764 TI - Mild tricuspid regurgitation: a benign fetal finding at various stages of gestation. PMID- 16374766 TI - Materials for peripheral nerve regeneration. AB - Recent efforts in scientific research in the field of peripheral nerve regeneration have been directed towards the development of artificial nerve guides. We have studied various materials with the aim of obtaining a biocompatible and biodegradable two layer guide for nerve repair. The candidate materials for use as an external layer for the nerve guides were poly(caprolactone) (PCL), a biosynthetic blend between PCL and chitosan (CS) and a synthesised poly(ester-urethane) (PU). Blending PCL, which is a biocompatible synthetic polymer, with a natural polymer enhanced the system biocompatibility and biomimetics, fastened the degradation rates and reduced the production costs. Various novel block poly(ester-urethane)s are being synthesised by our group with tailored properties for specific tissue engineering applications. One of these poly(ester-urethane)s, based on a low molecular weight poly(caprolactone) as the macrodiol, cycloesandimethanol as the chain extender and hexamethylene diisocyanate as the chain linker, was investigated for the production of melt extruded nerve guides. We studied natural polymers such as gelatin (G), poly(L lysine) (PL) and blends between chitosan and gelatin (CS/G) as internal coatings for nerve guides. In vitro and in vivo tests were performed on PCL guides internally coated either with G or PL to determine the differences in the quality of nerve regeneration associated with the type of adhesion protein. CS/G natural blends combined the good cell adhesion properties of the protein phase with the ability to promote nerve regeneration of the polysaccharide phase. Natural blends were crosslinked both by physical and chemical crosslinking methods. In vitro neuroblast adhesion tests were performed on CS/G film samples, PCL/CS and PU guides internally coated with G to evaluate the ability of such materials towards nerve repair. PMID- 16374767 TI - Lithium ion induced stabilization of the liquid crystalline DNA. AB - A comparative study of the effects of alkali metal ions Li(+), Na(+), K(+), Rb(+), and Cs(+) on the liquid crystalline organization of high-molecular-weight calf thymus DNA using polarized light microscopy was performed. Major differences in the behavior of Li(+) as compared to the other ions were found. Critical DNA concentration expected to exhibit anisotropic behavior was found to be the same for all the monovalent ions, except for Li(+). DNA initially showed cholesteric textures, which later changed to higher ordered columnar phase for all ions, with the cholesteric-columnar transition facilitated upon increasing the size of the counterion. For Li(+) ion, a nematic schlieren-like texture was formed initially, which after a few days changed to a highly stable (for more than 2 months) biphasic cholesteric-columnar arrangement. The observed differences between Li(+) and other alkali metal ions could be rationalized on the basis of the higher number of hydration water molecules of Li(+) and its complexation behavior. Highly stable DNA mesophases may find applications in the field of nanoelectronics, in designing biosensing units, and in DNA chips. PMID- 16374768 TI - Polymeric nanocapsules containing an antiseptic agent obtained by controlled nanoprecipitation onto water-in-oil miniemulsion droplets. AB - The modified nanoprecipitation of polymers onto stable nanodroplets has been successfully applied to prepare well-defined nanocapsules whose core is composing of an antiseptic agent, i.e., chlorhexidine digluconate aqueous solution. The stable nanodroplets were obtained by inverse miniemulsions with an aqueous antiseptic solution dispersed in an organic medium of solvent/nonsolvent mixture containing an oil-soluble surfactant and the polymer for the shell formation. The change of gradient of the solvent/nonsolvent mixture of dichloromethane/cyclohexane, obtained by heating at 50 degrees C, led to the precipitation of the polymer in the organic continuous phase and deposition onto the large interface of the aqueous miniemulsion droplets. The monodisperse polymer nanocapsules with the size range of 240-80 nm were achieved as a function of the amount of surfactant. Using various polymer contents, molecular weights and types, an encapsulation efficiency of 20-100% was obtained as detected by proton-nuclear magnetic resonance spectroscopy ((1)H NMR) measurements. The nanocapsules could be easily transferred into water as continuous phase resulting in aqueous dispersions with nanocapsules containing an aqueous core with the antiseptic agent. The encapsulated amount of the antiseptic agent was evaluated to indicate the durability of the nanocapsule's wall. In addition, the use of different types of polymers having glass transition temperatures (T(g)) ranging from 10 to 100 degrees C in this process has been also successful. PMID- 16374769 TI - Adsorption of biopolyester depolymerase on silicon wafer and poly[(R)-3 hydroxybutyric acid] single crystal revealed by real-time AFM. AB - The adsorption behavior of PHB depolymerase from R. pickettii T1 on a silicon wafer and on P(3HB) single crystals has been studied by real-time and AFM in air and a buffer solution. First, the morphology of PHB depolymerase adsorbed on a silicon wafer was characterized to show that one molecule of PHB depolymerase has dimensions of 2.2 +/- 0.7 nm height and 16 +/- 5 nm width. The observation of PHB depolymerase adsorbed on a P(3HB) single crystal indicated that the dimensions of enzyme on the crystalline surface in air were 1.2 +/- 0.5 nm high and 28 +/- 7 nm wide, while enzyme molecules with dimensions of 2.1 +/- 0.6 nm height and 16 +/- 7 nm width were detected in a buffer solution. Comparison of the dimensions of PHB depolymerase in air with those in a buffer solution showed that the enzyme was squashed in air, but not in a buffer solution. In addition, the influence of enzymatic adsorption on the molecular state of the P(3HB) crystalline surface was investigated. The AFM images of P(3HB) single crystals after enzymatic adsorption and washing with ethanol indicated that the adhesion of PHB depolymerase changed the molecular state and generated holes on the crystalline surface. PMID- 16374770 TI - Patterning of a random copolymer of poly[lactide-co-glycotide-co-(epsilon caprolactone)] by UV embossing for tissue engineering. AB - The random copolymer, poly[lactide-co-glycotide-co-(epsilon-caprolactone)] (PLGACL) diacrylate was prepared by ring-opening polymerization of L-lactide, glycolide, and epsilon-caprolactone initiated with tetra(ethylene glycol). The diacrylated polymers were extensively characterized. With a UV embossing method, these copolymers were successfully fabricated into microchannels separated by microwalls with a high aspect (height/width) ratio. The PLGACL network films showed good cytocompatibility. Varieties of microstructures were fabricated, such as 10 x 40 x 60, 10 x 80 x 60, 25 x 40 x 60, or 25 x 80 x 60 microm(3) structures (microwall width x microchannel width x microwall height). The results demonstrated that smooth muscle cells (SMCs) can grow not only on the microchannel surfaces but also on the surfaces of the microwall and sidewall. The SMCs aligned along the 25 microm wide microwall with an elongated morphology and proliferated very slowly in comparison to those on the smooth surface with a longer cell-culture term. Few cells could attach and spread on the surface of the 40 microm wide microchannel, while the cells flourished on the 80 microm, or more than 80 microm, wide microchannel with a spindle morphology. The biophysical mechanism mediated by the micropattern geometry is discussed. Overall, the present micropattern, consisting of biodegradable and cytocompatible PLGACL, provides a promising scaffold for tissue engineering. PMID- 16374771 TI - Triclosan release from coated polyglycolide threads. AB - Copolymerization of lactide, epsilon-caprolactone, and trimethylene carbonate is performed. The synthesis is focused on obtaining materials with adequate properties for application as a suture coating that could contain an antimicrobial agent like triclosan. An amorphous character, a low glass transition temperature, a moderate susceptibility to degradation, and a hydrophobic nature are the conditions required for the optimal behavior of this coating. These properties can be attained with a copolymer of composition 10:60:30. Triclosan is added to the surface of polyglycolide threads and its release is studied in different media with high-performance liquid chromatography. The influence of the temperature, the diameter of the thread, the initial concentration of the antibacterial agent, and the applied procedure on the incorporation of triclosan is also evaluated. A total release of triclosan is attained after a few days of exposure to a Dulbecco's based medium, whereas equilibrium concentrations are reached when a Sorensen hydrophilic medium is used. Partition and diffusion coefficients are also estimated. PMID- 16374772 TI - Preparation and characterization of novel bone scaffolds based on electrospun polycaprolactone fibers filled with nanoparticles. AB - Novel bone-scaffolding materials were successfully fabricated by electrospinning from polycaprolactone (PCL) solutions containing nanoparticles of calcium carbonate (CaCO(3)) or hydroxyapatite (HA). The diameters of the as-spun fibers were found to increase with the addition and increasing amounts of the nanoparticles. The observed increase in the diameters of the as-spun fibers with the addition and increasing amounts of the nanoparticulate fillers was responsible for the observed increase in the tensile strength of the obtained fiber mats. An increase in the concentration of the base PCL solution caused the average diameter of the as-spun PCL/HA composite fibers to increase. Increasing applied electrical potential also resulted in an increase in the diameters of the obtained PCL/HA composite fibers. Lastly, indirect cytotoxicity evaluation of the electrospun mats of PCL, PCL/CaCO(3), and PCL/HA fibers based on human osteoblasts (SaOS2) and mouse fibroblasts (L929) revealed that these as-spun mats posed no threat to the cells, a result that implied their potential for utilization as bone-scaffolding materials. PMID- 16374773 TI - Chemo-enzymatic synthesis of raffinose-branched polyelectrolytes and self assembly application in microcapsules. AB - A novel biocompatible polyelectrolyte poly(vinyl raffinose-co-acrylic acid) (PRCA) containing a raffinose branch was prepared via redox polymerization using Fe(2+)/K(2)S(2)O(8)/H(2)O(2) starting from enzymatically-synthesized monomer: 1-O vinyldecanedioyl raffinose. Copolymers with different monomer feed ratios were prepared and characterized with IR, NMR, and GPC. PRCA can be alternated with polycation to form microcapsules on a crystals template by electrostatic layer-by layer technique. The multilayers of PRCA/poly(methacryloyloxyethyl dimethylbenzyl ammonium chloride) (PMBA) on quartz slides and PRCA/poly(dimethyldiallyl ammonium chloride) (PDDA) on acyclovir crystals template were fabricated and characterized with UV-Vis spectra, the microelectrophoretic measurement, and TEM. Hollow capsules can be formed after the removal of acyclovir crystals template in a buffer solution. The nano-capsule-carrying galactose residue is a potential targeting drug-controlled delivery systems. PMID- 16374774 TI - Homogenous synthesis of hydroxyethylcellulose in NaOH/urea aqueous solution. AB - Hydroxyethylcellulose (HEC) was synthesized by a fully homogenous method from cellulose in 7.5 wt.-% NaOH/11 wt.-% urea aqueous solutions under mild conditions. HEC samples were characterized with NMR, SEC-LLS, solubility, and viscosity measurements. The MS and DS values of the obtained HEC samples are in the range from 0.54 to 1.44 and 0.45 to 1.14, respectively, and the relative DS values at C-2 and C-6 hydroxyl groups are slightly higher than those at C-3 hydroxyl groups. HEC samples are soluble in water starting from a MS of 0.57 and DS of 0.49, which display high viscosity in aqueous solutions. Moreover, a NaOH/urea aqueous solution is a stable system for cellulose etherification. In this way, we could provide a simple, pollution-free, and homogeneous aqueous solution system for synthesizing cellulose ethers. PMID- 16374775 TI - Promotion of cell affinity of porous PLLA scaffolds by immobilization of RGD peptides via plasma treatment. AB - In the present work, RGDS (Arg-Gly-Asp-Ser) was immobilized on PLLA scaffolds with plasma treatment. The amount of immobilization, determined by HPLC, was confirmed to be in the effective order. Results from the culture of rat osteosarcoma (ROS), osteoblastic-like cells, demonstrate that the immobilization of RGDS could effectively enhance the attachment of ROS cells on PLLA and increase the cell density in PLLA scaffolds. In addition, experiments of in vitro mineralization indicate that there were more cells and mineralization focci in the RGDS-immobilized scaffolds, suggesting a tendency to form bone-like tissues, compared with the unmodified PLLA scaffold. On the other hand, the PLLA scaffolds immobilized with RGES (Arg-Gly-Glu-Ser) were much less effective in promotion of ROS attachment, suggesting that the enhancement on cell attachment was mainly due to the recognition of RGDS by the adhesion receptors on the cell membrane. The results presented in this work demonstrate that RGDS could be successfully immobilized on PLLA scaffolds with plasma treatment and such modification can make PLLA scaffolds more suitable for culture of osteoblast-like cells and for generation of bone-like tissues. PMID- 16374776 TI - DNA-based gels for oral delivery of probiotic bacteria. AB - A single-stranded DNA, readily extracted from industrial discarded salmon milt, was used to prepare hydrogels and complex gels by cross-linking with gelatin and kappa-carrageenan, for the oral delivery of probiotic bacteria. The complex gels showed a higher protective capability over the hydrogels for approximately one log scale. However, the hydrogels were more stable during storage at 4 degrees C. The Lactobacillus and Lactococcus due to protection of the hydrogels could better tolerate to acid than the Bifidobacterium. Furthermore, food-graded hydrogels were prepared and optimized to a similar protective capability for future applications. PMID- 16374778 TI - Non-PKC DAG/phorbol-ester receptor(s) inhibit complement receptor-3 and nPKC inhibit scavenger receptor-AI/II-mediated myelin phagocytosis but cPKC, PI3k, and PLCgamma activate myelin phagocytosis by both. AB - Complement-receptor-3 (CR3/MAC-1), scavenger-receptor-AI/II (SRAI/II), and Fcgamma-receptor (FcgammaR) can mediate myelin phagocytosis in macrophages and microglia. Paradoxically, after injury to CNS axons these receptors are expressed but myelin is not phagocytosed, suggesting that phagocytosis is subject to regulation between efficient and inefficient states. In the present work, we focus on CR3/MAC-1 and SRAI/II-mediated myelin phagocytosis. Phagocytosis by CR3/MAC-1 and SRAI/II was inhibited by cPKC inhibitor Go-6976, general-PKC inhibitors Ro-318220 and calphostin-C, and BAPTA/AM, which chelates intracellular Ca2+ required for cPKC activation. Signaling/activation by cPKC are thus suggested. PMA, which mimics diacylglycerol (DAG) as an activator of cPKC, novel PKC (nPKC), and non-PKC DAG-driven molecule(s), produced a dose-dependent dual effect on phagocytosis by CR3/MAC-1 and SRAI/II, i.e., augmentation at low concentrations and inhibition at high concentrations. Inhibition of phagocytosis by CR3/MAC-1 was enhanced by combining inhibiting concentrations of PMA with PKC inhibitors Go-6976 or Ro-318220, suggesting inhibition by PMA/DAG-driven non-PKC molecule(s). In contrast, inhibition of phagocytosis by SRAI/II was enhanced by combining inhibiting concentrations of PMA with cPKC inhibitor Go-6976 but not with general-PKC inhibitor Ro-318220, suggesting inhibition by nPKC. Phagocytosis by CR3/MAC-1 and SRAI/II was further inhibited by PI3K inhibitors wortmannin and LY-294002 and PLCgamma inhibitor U-73122. Altogether, our observations suggest that CR3/MAC-1 and SRAI/II-mediated myelin phagocytosis share activation by PI3K, PLCgamma and cPKC. The two differ, however, in that non-PKC DAG-driven molecule(s) inhibit CR3/MAC-1-mediated phagocytosis, whereas nPKC inhibit SRAI/II mediated phagocytosis. Each of these signaling steps may be targeted for regulating CR3/MAC-1 and/or SRAI/II-mediated phagocytosis between efficient and inefficient states. PMID- 16374777 TI - Upregulation of the stress-associated gene p8 in mouse models of demyelination and in multiple sclerosis tissues. AB - Cuprizone-induced demyelination is a mouse model of multiple sclerosis (MS) as cuprizone-fed mice exhibit neuroinflammation and demyelination in the brain. Upon removal of cuprizone from the diet, inflammation is resolved and reparative remyelination occurs. In an Affymetrix GeneChip analysis, the stress-associated gene p8 was strongly upregulated (>10x) during cuprizone-induced demyelination but not remyelination. We verified this upregulation (>15x) of p8 in the CNS during demyelination by real-time polymerase chain reaction (PCR). This upregulation is brain-specific, as p8 is not elevated in the liver, lung, kidney, spleen, and heart of cuprizone-treated mice. We also localized the cellular source of p8 during cuprizone treatment, and further found elevated expression during embryogenesis but not in normal adult brain. Compared with wild-type controls, the death of oligodendrocytes in p8-/- mice is delayed, as is microglial recruitment to areas of demyelination. The corpus callosum of p8-/- mice demyelinates at a slower rate than wild-type mice, suggesting that p8 exacerbates CNS inflammation and demyelination. Enhanced expression of p8 is also observed in the spinal cords of mice with acute experimental autoimmune encephalomyelitis (EAE) induced by PLP139-151 peptide (10x). Increased expression is detected during disease onset and expression wanes during the remission phase. Finally, p8 is found upregulated (8x) in post-mortem tissue from MS patients and is higher in the plaque tissue compared with adjacent normal-appearing white and gray matter. Thus, p8 is an excellent candidate as a novel biomarker of demyelination. PMID- 16374779 TI - Neuroprotective actions of endogenous interleukin-1 receptor antagonist (IL-1ra) are mediated by glia. AB - The pro-inflammatory cytokine interleukin-1 (IL-1), contributes to neuronal inflammation and cell death induced by ischemia, excitotoxicity, or trauma, while administration of IL-1 receptor antagonist (IL-1ra) reduces neuronal injury. The aim of the present study was to test the hypothesis that endogenous IL-1ra is neuroprotective in vivo and in vitro, and to identify its mechanism of actions. Mice lacking IL-1ra (IL-1ra knock-out (KO]) exhibited a dramatic increase in neuronal injury (3.6-fold increase in infarct size) induced by transient cerebral ischemia compared to wild-type (WT) animals. Basal cell death of cultured cortical neurons from WT and IL-1ra KO was identical, and treatment with NMDA or AMPA (20 microM) increased cell death to the same extent in WT and IL-1ra KO neurons. However, basal and NMDA- or AMPA-induced cells death was significantly higher in glial-neuronal co-cultures from IL-1ra KO than from WT mice. We further showed that pure microglial cultures, but not pure astrocytes cultures, released IL-1ra in response to treatment with conditioned medium from NMDA- or AMPA treated primary neurons. These results demonstrate that endogenous IL-1ra produced by microglia is neuroprotective in cerebral ischemia or excitotoxicity. PMID- 16374780 TI - Selectively increased expression of the astrocytic/endothelial glucose transporter protein GLUT1 in acute liver failure. AB - Acute liver failure (ALF) is consistently accompanied by alterations in brain energy metabolites and recent nuclear magnetic resonance (NMR) studies suggest disturbances in brain oxidative metabolism in experimental ALF. Glucose transport across the blood-brain barrier is essential to sustain brain energy metabolism and is accomplished by the facilitative glucose transporter GLUT1. To investigate alterations in brain glucose uptake in acute liver failure further, GLUT1 expression and [14C]-2-deoxy-D-glucose uptake were measured in the brains of rats with hepatic devascularization. RT-PCR and Western blot analyses showed significant increases in steady-state levels of GLUT1 mRNA and protein in frontal cortex as early as 6 h following hepatic devascularization, (prior to the onset of brain edema and encephalopathy) which remained elevated at coma stages of encephalopathy. Expression of the astrocytic (45-kDa) and endothelial (55-kDa) forms of GLUT1 was increased as a result of hepatic devascularization. Exposure of cultured astrocytes to pathophysiologically relevant concentrations of ammonia resulted in increased GLUT1 expression, suggesting that elevated ammonia levels are responsible for GLUT1 upregulation in ALF. Increased GLUT1 expression in ALF was selective, since expression of the neuronal glucose transporter GLUT3 and other glucose-regulated proteins (GRP-78 and GRP-94) was unaltered. [14C]-2-deoxy D-glucose autoradiography revealed increases in cerebral glucose uptake following the induction of GLUT1 in ALF. These results suggest that ammonia-induced increases of GLUT1 expression resulting in increased cerebral glucose uptake occur in ALF and could contribute to the pathophysiological mechanisms responsible for the neurological complications of this condition. PMID- 16374781 TI - Comparison study of a traditional pulsed dye laser versus a long-pulsed dye laser in the treatment of early childhood hemangiomas. AB - BACKGROUND AND OBJECTIVE: The role of pulsed dye laser (PDL) in the treatment of childhood hemangiomas is still controversial because of the inherent characteristics of hemangiomas as well as the side effects of the PDL. Recently, the long pulsed dye laser (LPDL) with cryogen spray cooling (CSC) has been found relatively more effective and safer than the PDL in the treatment of port-wine stains and telangiectasia. This study was designed to compare the efficacy and complication rate of PDL versus LPDL for treating childhood hemangiomas. STUDY DESIGN/MATERIALS AND METHODS: We did a prospective, randomized, controlled trial in which we enrolled 52 Asian infants, aged 1-3 months, with early hemangiomas. These infants were assigned to PDL treatment (n = 26) or LPDL treatment (n = 26) and followed to age 1 year. A PDL with a wavelength of 585 nm and spot size of 7 mm and a LPDL with a wavelength of 595 nm and spot size of 7 mm were used. Each patient in the PDL group was treated with energy fluence between 6 and 7 J/cm(2) and a pulse duration of 0.45 milliseconds without epidermal cooling. Each patient in the LPDL group was treated with energy fluence between 9 and 15 J/cm(2) and a pulse duration of 10-20 milliseconds, utilizing CSC to protect the epidermis. Each group was treated at 4-week intervals until the lesion cleared. When each patient reached an age of 1 year, outcome measures such as clearance rate, time period of maximum proliferation, and complications were assessed. RESULTS: The number of children whose lesions showed complete clearance or minimal residual signs at 1 year of age was 14 (54%) in the PDL group and 17 (65%) in the LPDL group (P = 0.397). Compared with the LPDL, PDL treated children had more hypopigmentation (3, 12% vs. 8, 31%; P = 0.001), more hyperpigmentation (2, 8% vs. 4, 15%; P = 0.005), and more textural changes (1, 4% vs. 6, 23%; P = 0.001). The average time period of maximum proliferation in the LPDL group was significantly shorter than that of the PDL group (106 days vs. 177 days; P = 0.01). CONCLUSION: Early treatment of childhood hemangiomas with the LPDL is safer and more effective than the PDL. PMID- 16374782 TI - NMR structure of the conserved novel-fold protein TA0743 from Thermoplasma acidophilum. PMID- 16374783 TI - A topology-constrained distance network algorithm for protein structure determination from NOESY data. AB - This article formulates the multidimensional nuclear Overhauser effect spectroscopy (NOESY) interpretation problem using graph theory and presents a novel, bottom-up, topology-constrained distance network analysis algorithm for NOESY cross peak interpretation using assigned resonances. AutoStructure is a software suite that implements this topology-constrained distance network analysis algorithm and iteratively generates structures using the three dimensional (3D) protein structure calculation programs XPLOR/CNS or DYANA. The minimum input for AutoStructure includes the amino acid sequence, a list of resonance assignments, and lists of 2D, 3D, and/or 4D-NOESY cross peaks. AutoStructure can also analyze homodimeric proteins when X-filtered NOESY experiments are available. The quality of input data and final 3D structures is evaluated using recall, precision, and F-measure (RPF) scores, a statistical measure of goodness of fit with the input data. AutoStructure has been tested on three protein NMR data sets for which high-quality structures have previously been solved by an expert, and yields comparable high-quality distance constraint lists and 3D protein structures in hours. We also compare several protein structures determined using AutoStructure with corresponding homologous proteins determined with other independent methods. The program has been used in more than two dozen protein structure determinations, several of which have already been published. PMID- 16374785 TI - Kinetics and mechanism of the electrooxidation of formic acid- spectroelectrochemical studies in a flow cell. PMID- 16374784 TI - Finite-size, fully addressable DNA tile lattices formed by hierarchical assembly procedures. PMID- 16374786 TI - A simple protocol to estimate differences in protein binding affinity for enantiomers without prior resolution of racemates. PMID- 16374787 TI - Vitamin B12: a methyl group without a job? PMID- 16374788 TI - NMR characterization of kinase p38 dynamics in free and ligand-bound forms. PMID- 16374789 TI - Production of liquid hydrocarbons from biomass. PMID- 16374790 TI - Supramolecular bioinorganic hybrid catalysts for enantioselective transformations. PMID- 16374791 TI - Stereoselective synthesis of oligo-alpha(2,8)-3-deoxy-D-manno-2-octulosonic acid derivatives. PMID- 16374792 TI - Remarkable amplification of the self-disproportionation of enantiomers on achiral phase chromatography columns. PMID- 16374793 TI - Composition of perineuronal nets in the adult rat cerebellum and the cellular origin of their components. AB - The decrease in plasticity that occurs in the central nervous system during postnatal development is accompanied by the appearance of perineuronal nets (PNNs) around the cell body and dendrites of many classes of neuron. These structures are composed of extracellular matrix molecules, such as chondroitin sulfate proteoglycans (CSPGs), hyaluronan (HA), tenascin-R, and link proteins. To elucidate the role played by neurons and glial cells in constructing PNNs, we studied the expression of PNN components in the adult rat cerebellum by immunohistochemistry and in situ hybridization. In the deep cerebellar nuclei, only large excitatory neurons were surrounded by nets, which contained the CSPGs aggrecan, neurocan, brevican, versican, and phosphacan, along with tenascin-R and HA. Whereas both net-bearing neurons and glial cells were the sources of CSPGs and tenascin-R, only the neurons expressed the mRNA for HA synthases (HASs), cartilage link protein, and link protein Bral2. In the cerebellar cortex, Golgi neurons possessed PNNs and also synthesized HASs, cartilage link protein, and Bral2 mRNAs. To see whether HA might link PNNs to the neuronal cell surface by binding to a receptor, we investigated the expression of the HA receptors CD44, RHAMM, and LYVE-1. No immunolabelling for HA receptors on the membrane of net bearing neurons was found. We therefore propose that HASs, which can retain HA on the cell surface, may act as a link between PNNs and neurons. Thus, HAS and link proteins might be key molecules for PNN formation and stability. PMID- 16374794 TI - Estrogen receptor-alpha distribution in male rodents is associated with social organization. AB - It has been hypothesized that site-specific reduction of estrogen receptor-alpha (ERalpha) is associated with the expression of male prosocial behaviors. Specifically, highly social males are predicted to express significantly lower levels of ERalpha than females and less social males in brain regions associated with prosocial behavior including the bed nucleus of the stria terminalis (BST) and the medial amygdala (MeA). This hypothesis was tested by comparing ERalpha immunoreactivity (IR) in three species of microtines, the polygynous montane (Microtus montanus) and meadow (M. pennsylvanicus) voles and the monogamous pine vole (M. pinetorum), and two species of cricetines that differ in the extent of social pair-bond formation, Siberian (Phodopus sungorus) and Djungarian (P. campbelli) hamsters. As predicted, ERalpha-IR was sexually dimorphic in the BST and MeA of the highly social species, with females expressing more ERalpha-IR cells than males. Male and female montane voles did not differ. Male and female meadow voles differed in the ventromedial hypothalamus, with females expressing more ERalpha-IR cells. Male pine voles expressed lower levels of ERalpha-IR in the MeA than male montane and meadow voles and in the BST relative to montane males. Male Djungarian hamsters, which show higher levels of parental care, had fewer ERalpha-IR cells in the BST than male Siberian hamsters. Results indicate that the distribution of ERalpha differs relative to the continuum of species typical affiliative behavior and supports the hypothesis that ERalpha has a significant role in regulating species-specific social organization. PMID- 16374795 TI - A phylotypic stage in vertebrate brain development: GABA cell patterns in zebrafish compared with mouse. AB - A recent comparison of early forebrain gene expression in mouse and zebrafish revealed highly comparable expression patterns of developmentally relevant genes, for example, of proneural (Neurogenin1, NeuroD, Mash1/Zash1a) genes involved in neurogenesis at a particular time window (mouse: embryonic day 12.5/13.5; zebrafish: 3 days). Here we extend this analysis to the description of gamma aminobutyric acid (GABA) cell patterns in the early postembryonic zebrafish brain (i.e., during early secondary neurogenesis). We find again an astonishing degree of correspondences of GABA cell patterns between zebrafish and mouse during this previously established critical time window, for example, regarding absence of GABA cells in certain forebrain regions (pallium, dorsal thalamus, eminentia thalami) or with respect to the spatiotemporal occurrence of GABA cells (e.g., late cerebellar GABA cells). Furthermore, there is perfect correlation with previously established proneural gene expression patterns (i.e., absence of Mash1/Zash1a gene expression in GABA-cell-free forebrain regions) between mouse and zebrafish. The available information in additional vertebrate species, especially in Xenopus, is also highly consistent with our analysis here and suggests that a "phylotypic stage" of neurogenesis during vertebrate brain development may be present. PMID- 16374798 TI - Mapping glutamate responses in immunocytochemically identified neurons of the mouse retina. AB - The mammalian retina contains as many as 50-60 unique cell types, many of which have been identified using various neurochemical markers. Retinal neurons express N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxyl-5-methylisoxazole-4-propionic acid (AMPA), and kainic acid (KA) receptor subunits in various mixtures, densities, and spatial distributions. Ionotropic glutamatergic drive in retinal neurons can be mapped using a cation channel permeant guanidinium analog called agmatine (1-amino-4-guanidobutane; AGB). This alternative approach to physiologically characterize neurons in the retina was introduced by Marc (1999, J Comp Neurol 407:47-64, 407:65-76), and allows the simultaneous mapping of responses of glutamate receptor-gated channels from an entire population of neurons. Unlike previous AGB studies, we colocalized AGB with various macromolecular markers using direct and indirect immunofluorescence to characterize the glutamate agonist sensitivities of specific cell types. Activation with NMDA, AMPA, and KA resulted in AGB entry into neurons in a dose dependent manner and was consistent with previous receptor subunit localization studies. Consistent with the various morphological phenotypes encompassed by the calbindin and calretinin immunoreactive cells, we observed various functional phenotypes revealed by AGB labeling. Not all calbindin or calretinin immunoreactive cells showed ligand-evoked AGB permeation. A small proportion either did not possess functional glutamate receptors, required higher activation thresholds, or express functional channels impermeable to AGB. AMPA and KA activation of bipolar cells resulted in AGB permeation into the hyperpolarizing variety only. We also studied the glutamate ligand-gating properties of 3[alpha1 3]-fucosyl-N-acetyl-lactosamine (CD15) immunoreactive cells and show functional responses consistent with receptor subunit gene expression patterns. CD15 immunoreactive bipolar cells only responded to AMPA but not KA. The CD15 immunoreactive amacrine cells demonstrated an identical selectivity to AMPA activation, but were also responsive to NMDA. Finally, localization of AGB secondary to glutamate receptor activation was visualized with a permanent reaction product. PMID- 16374796 TI - Water deprivation activates a glutamatergic projection from the hypothalamic paraventricular nucleus to the rostral ventrolateral medulla. AB - Elevated sympathetic outflow contributes to the maintenance of blood pressure in water-deprived rats. The neural circuitry underlying this response may involve activation of a pathway from the hypothalamic paraventricular nucleus (PVH) to the rostral ventrolateral medulla (RVLM). We sought to determine whether the PVH RVLM projection activated by water deprivation is glutamatergic and/or contains vasopressin- or oxytocin-neurophysins. Vesicular glutamate transporter 2 (VGLUT2) mRNA was detected by in situ hybridization in the majority of PVH neurons retrogradely labeled from the ipsilateral RVLM with cholera toxin subunit B (CTB; 85% on average, with regional differences). Very few RVLM-projecting PVH neurons were immunoreactive for oxytocin- or vasopressin-associated neurophysin. Injection of biotinylated dextran amine (BDA) into the PVH produced clusters of BDA-positive nerve terminals within the ipsilateral RVLM that were immunoreactive (ir) for the VGLUT2 protein. Some of these terminals made close appositions with tyrosine-hydroxylase-ir dendrites (presumptive C1 cells). In water-deprived rats (n=4), numerous VGLUT2 mRNA-positive PVH neurons retrogradely labeled from the ipsilateral RVLM with CTB were c-Fos-ir (16-40%, depending on PVH region). In marked contrast, few glutamatergic, RVLM-projecting PVH neurons were c-Fos-ir in control rats (n=3; 0-3%, depending on PVH region). Most (94% +/- 4%) RVLM projecting PVH neurons activated by water deprivation contained VGLUT2 mRNA. In summary, most PVH neurons that innervate the RVLM are glutamatergic, and this population includes the neurons that are activated by water deprivation. One mechanism by which water deprivation may increase the sympathetic outflow is activation of a glutamatergic pathway from the PVH to the RVLM. PMID- 16374799 TI - Trafficking of macromolecules and organelles in cultured Dystonia musculorum sensory neurons is normal. AB - Dystonia musculorum (dt) mice suffer from a recessive neuropathy characterized by the progressive loss of sensory axons. The gene responsible for this disorder, dystonin/Bpag1, encodes several alternatively spliced forms of a cytoskeletal linker protein. Neural isoforms of dystonin/Bpag1 are predicted to link actin filaments to microtubules. Consistent with this, previous observations have demonstrated that the cytoskeleton within sensory neurites of dt mice is perturbed. Also, recent results have indicated that a neural isoform of dystonin/Bpag1 interacts with the dynein motor complex. Because microtubule organization and dynein motor function are essential for trafficking, we hypothesized that this process would be perturbed in dt sensory neurons. Here, we demonstrate that cultured primary dorsal root ganglion (DRG) neurons express dystonin/Bpag1 and that loss of this expression causes an increase in apoptosis and a decrease in average neurite length. In contrast, detailed examination showed that the organization of microtubules is indistinguishable in DRG neuronal cultures from neonatal dt and wild-type mice. In addition, the steady-state distribution of several molecules and organelles is unchanged in these cultures. Furthermore, the speeds of mitochondrial movement in both anterograde and retrograde directions were comparable in dt and wild-type sensory neurons cultured from neonatal mice. Thus, dystonin/Bpag1 is not essential for microtubule network assembly since the microtubule network is intact in short term cultures of sensory neurons from neonatal mice lacking this protein. In addition, dystonin/Bpag1 is not an essential part of the dynein motor complex for mitochondrial transport since mitochondrial trafficking is normal in cultured sensory neurons from dt mice. PMID- 16374801 TI - Neural activity- and development-dependent expression and distribution of CASK interacting nucleosome assembly protein in mouse brain. AB - CASK interacting nucleosome assembly protein (CINAP) modulates gene expression and its abundance in cultured neurons is regulated by synaptic activity. To further study the function of CINAP in vivo, we examined the temporal and spatial expression profiles of CINAP. CINAP was widely expressed in different regions of adult mouse brain, including the cerebral cortex, hippocampus, striatum, hypothalamus, cerebellum, and two adult brain regions known to generate progenitor neurons. During early development, CINAP was also expressed in regions where neuronal progenitor cells were actively dividing, the ventricular and subventricular zones, suggesting that in addition to regulating gene expression in mature neurons, CINAP may also play a role in dividing cells. Since the hypothalamus responds to several physiological responses, we examined whether CINAP protein levels in the paraventricular nucleus (PVN) of the hypothalamus are regulated by changes in osmolality achieved through oral administration of hypertonic saline. Compared with control mice, mice treated with hypertonic saline expressed higher CINAP protein levels in the PVN, supporting a role of CINAP in neural response in vivo. Using confocal microscopic analysis, a significant amount of CINAP was found in the cytoplasm of neurons. Biochemical fractionation further confirmed that CINAP was associated with synapses, suggesting a translocation of CINAP protein from synapse to the nucleus. Consistent with this speculation, nuclear CINAP levels in the PVN were higher in hypertonic saline-treated mice than those who drank water. CINAP may be regulated through changes in protein stability and nuclear translocation in neurons. PMID- 16374800 TI - Comparative analysis of lesion development and intraspinal inflammation in four strains of mice following spinal contusion injury. AB - Susceptibility to neuroinflammatory disease is influenced in part by genetics. Recent data indicate that survival of traumatized neurons is strain dependent and influenced by polygenic loci that control resistance/susceptibility to experimental autoimmune encephalomyelitis (EAE), a model of CNS autoimmune disease. Here, we describe patterns of neurodegeneration and intraparenchymal inflammation after traumatic spinal cord injury (SCI) in mice known to exhibit varying degrees of EAE susceptibility [EAE-resistant (r) or EAE-susceptible (s) mice]. Spinal cords from C57BL/6 (EAE-s), C57BL/10 (EAE-r), BALB/c (EAE-r), and B10.PL (EAE-s) mice were prepared for stereological and immunohistochemical analysis at 6 hours or 3, 7, 14, 28, or 42 days following midthoracic (T9) spinal contusion injury. In general, genetic predisposition to EAE predicted the magnitude of intraparenchymal inflammation but not lesion size/length or locomotor recovery. Specifically, microglia/macrophage activation, recruitment of neutrophils and lymphocytes, and de novo synthesis of MHC class II were greatest in C57BL/6 mice and least in BALB/c mice at all times examined. However, lesion volume and axial spread of neurodegeneration were similar in C57BL/6 and BALB/c mice and were significantly greater than in C57BL/10 or B10.PL mice. Strains with marked intraspinal inflammation also developed the most intense lesion fibrosis. Thus, strain-dependent neuroinflammation was observed after SCI, but without a consistent relationship to EAE susceptibility or lesion progression. Only in C57BL/6 mice was the magnitude of intraspinal inflammation predictive of secondary neurodegeneration, functional recovery, or fibrosis. PMID- 16374803 TI - AII amacrine cells in the distal inner nuclear layer of the mouse retina. AB - We serendipitously found a distal Disabled-1 (Dab1)-immunoreactive cell in retina of the C57BL/6J black mouse. The somata of these cells are located in the outermost part of the inner nuclear layer (INL). Their processes extend toward the outer plexiform layer (OPL), receiving synaptic inputs from horizontal and interplexiform cells. In the current study, we name this cell the "distal Dab1 immunoreactive cell." Double-labeling experiments demonstrate that the distal Dab1-immunoreactive cell is not a horizontal cell. Rather, the distal Dab1 cell appears to be a misplaced AII cell, by being glycine transporter-1-immunoreactive and by resembling the latter cell in an electron microscopic analysis. A distal Dab1 cell had been reported in the FVB/N mouse retina, a model of retinitis pigmentosa (Park et al. [2004] Cell Tissue Res 315:407-412). However, here, we found this distal Dab1-immunoreactive cell in the adult and normal developing mouse retinas. Hence, we show that such cells do not require the loss of photoreceptors as suggested previously (Park et al. [2004] Cell Tissue Res 315:407-412). Instead, two other pieces of data suggest an alternative explanation sources for distal Dab1 cells. First, we find a correlation between the number of these cells in the left and right eyes Second, developmental analysis shows that the distal Dab1-immunoreactive cell is first observed shortly after birth. At the same time, AII cells emerge, extending their neurites into the inner retina. These data suggest that distal Dab1-immunoreactive cells are misplaced AII amacrine cells, resulting from genetically modulated anomalies owing to migration errors. PMID- 16374802 TI - Pyramidal cells of the rat basolateral amygdala: synaptology and innervation by parvalbumin-immunoreactive interneurons. AB - The generation of emotional responses by the basolateral amygdala is determined largely by the balance of excitatory and inhibitory inputs to its principal neurons, the pyramidal cells. The activity of these neurons is tightly controlled by gamma-aminobutyric acid (GABA)-ergic interneurons, especially a parvalbumin positive (PV(+)) subpopulation that constitutes almost half of all interneurons in the basolateral amygdala. In the present semiquantitative investigation, we studied the incidence of synaptic inputs of PV(+) axon terminals onto pyramidal neurons in the rat basolateral nucleus (BLa). Pyramidal cells were identified by using calcium/calmodulin-dependent protein kinase II (CaMK) immunoreactivity as a marker. To appreciate the relative abundance of PV(+) inputs compared with excitatory inputs and other non-PV(+) inhibitory inputs, we also analyzed the proportions of asymmetrical (presumed excitatory) synapses and symmetrical (presumed inhibitory) synapses formed by unlabeled axon terminals targeting pyramidal neurons. The results indicate that the perisomatic region of pyramidal cells is innervated almost entirely by symmetrical synapses, whereas the density of asymmetrical synapses increases as one proceeds from thicker proximal dendritic shafts to thinner distal dendritic shafts. The great majority of synapses with dendritic spines are asymmetrical. PV(+) axon terminals form mainly symmetrical synapses. These PV(+) synapses constitute slightly more than half of the symmetrical synapses formed with each postsynaptic compartment of BLa pyramidal cells. These data indicate that the synaptology of basolateral amygdalar pyramidal cells is remarkably similar to that of cortical pyramidal cells and that PV(+) interneurons provide a robust inhibition of both the perisomatic and the distal dendritic domains of these principal neurons. PMID- 16374804 TI - Ommatidial type-specific interphotoreceptor connections in the lamina of the swallowtail butterfly, Papilio xuthus. AB - The eye of the butterfly Papilio xuthus contains a random array of three types of ommatidia (types I-III), each bearing nine photoreceptors, R1-R9. Of the six spectral classes of photoreceptors identified, types I, II, and III ommatidia contain four, three, and two classes, respectively: the ommatidia are thus spectrally heterogeneous. The photoreceptors send their axons to the lamina where, together with some large monopolar cells (LMCs), the nine from a single ommatidium contribute to a module called a lamina cartridge. We recently reported that among different photoreceptor axon terminals visualized by confocal microscopy, the number and length of axon collaterals differ for different spectral receptors, suggesting that lamina circuits are specific for each ommatidial type. Here we studied the distribution of synapse-like structures in the cartridges, first characterizing a photoreceptor by measuring its spectral sensitivity and then injecting Lucifer yellow (LY). We subsequently histologically identified the type of ommatidium to which the injected photoreceptor belonged, cut serial ultrathin sections of the entire lamina, labeled these with anti-LY immunocytochemistry, and then localized synapse-like structures. We found numerous interphotoreceptor contacts both within and between cartridges, the combination of which was again specific for the ommatidial type. R3 and R4, which are green-sensitive photoreceptors in all ommatidia, have thick axons lacking collaterals. We found that these cells exclusively make contacts with LMCs and not with photoreceptors. We therefore presume that R3 and R4 construct a system for motion vision, whereas other randomly distributed spectral types provide inputs for color vision. PMID- 16374805 TI - Family-based association test for time-to-onset data with time-dependent differences between the hazard functions. AB - In genetic association studies, the differences between the hazard functions for the individual genotypes are often time-dependent. We address the non proportional hazards data by using the weighted logrank approach by Fleming and Harrington [1981]:Commun Stat-Theor M 10:763-794. We introduce a weighted FBAT Logrank whose weights are based on a non-parametric estimator for the genetic marker distribution function under the alternative hypothesis. We show that the computation of the marker distribution under the alternative does not bias the significance level of any subsequently computed FBAT-statistic. Hence, we use the estimated marker distribution to select the Fleming-Harrington weights so that the power of the weighted FBAT-Logrank test is maximized. In simulation studies and applications to an asthma study, we illustrate the practical relevance of the new methodology. In addition to power increases of 100% over the original FBAT Logrank test, we also gain insight into the age at which a genotype exerts the greatest influence on disease risk. PMID- 16374806 TI - Incorporating endophenotypes into allele-sharing based linkage tests. AB - For a genetic study in which there are concordant and discordant sibpairs for a complex disease trait and the measurements of other endophenotypes/intermediate phenotypes for each of the individuals are also available, we describe an allele sharing based multipoint linkage test that utilizes nonparametrically the additional endophenotypes/intermediate phenotypes. The usefulness of this method is evaluated in simulation studies, which show that the gain in power is influenced by not only the endophenotypic value but also the correlation between the diagnosis-based phenotype and the endophenotype. In addition to reporting p values, our method also provides an index C(E), derived from the coefficients of the weight function associated with the endophenotype in the proposed statistic, to indicate the relevance of a specific endophenotype/intermediate phenotype in the genetic study. The simulation study indicates that a larger power, in general, corresponds to a larger value of the index C(E). The index C(E) is thus suggested as a quantity to be used in the choice of endophenotypes in linkage study. Data from the Stanford Asian Pacific Program in Hypertension and Insulin Resistance (SAPPHIRe) are used to illustrate the method. PMID- 16374807 TI - Localized brain activation specific to auditory memory in a female songbird. AB - Song acquisition in songbird males is a prominent model system for the study of the brain mechanisms of memory. Male zebra finches (Taeniopygia guttata) learn their songs from an adult conspecific tutor early in life. Previous work has shown that exposure of males to their tutor song leads to increased expression of immediate early genes (IEGs) in the caudomedial nidopallium (NCM) and in the caudomedial mesopallium (CMM). In addition, IEG expression in the NCM correlates significantly with the strength of song learning. Interpretation of these findings is complicated, as males both learn the characteristics of tutor song and learn to produce a similar own song. Female zebra finches do not sing, but nevertheless they learn the characteristics of a song to which they were exposed when young, and form a preference for it. Here, adult zebra finch females reared with their fathers showed a significant preference for their father's song. Females that were later reexposed to their father's song showed significantly greater expression of Zenk, the protein product of the IEG ZENK, than controls that were exposed to a novel song, in the CMM, but not in the NCM or hippocampus. These results suggest that in female zebra finches the CMM may be (part of) the neural substrate for the representation of the memory of their father's song. PMID- 16374810 TI - On-line confocal imaging of the events leading to structural dedifferentiation of an axonal segment into a growth cone after axotomy. AB - The transformation of a transected axonal tip into a growth cone (GC) after axotomy is a critical step in the cascade of events leading to regeneration. However, the mechanisms underlying it are largely unknown. In earlier studies we reported that axotomy of cultured Aplysia neurons leads to a transient and local increase in the free intracellular Ca2+ concentration, calpain activation, and localized proteolysis of the submembranal spectrin. In a recent ultrastructural study, we reported that calpain activation is critical for the restructuring of the microtubules and neurofilaments at the cut axonal end to form a compartment in which vesicles accumulate. By using on-line confocal imaging of microtubules (MTs), actin, and vesicles in cultured Aplysia neurons, we studied the kinetics of the transformation and examined some of the mechanisms that orchestrate it. We report that perturbation of the MTs' polymerization by nocodazole inhibits the formation of an MT-based compartment in which the vesicles accumulate, yet actin repolymerization proceeds normally to form a nascent GC's lamellipodium. Nevertheless, under these conditions, the lamellipodium fails to expand and form neurites. When actin filament polymerization is inhibited by cytochalasin D or jasplakinolide, the MT-based compartment is formed and vesicles accumulate at the cut axonal end. However, a GC's lamellipodium is not formed, and the cut axonal end fails to regenerate. A growth-competent GC is formed only when MT restructuring, the accumulation of vesicles, and actin polymerization properly converge in time and space. PMID- 16374808 TI - Abnormal expression of the G-protein-activated inwardly rectifying potassium channel 2 (GIRK2) in hippocampus, frontal cortex, and substantia nigra of Ts65Dn mouse: a model of Down syndrome. AB - Ts65Dn, a mouse model of Down syndrome (DS), demonstrates abnormal hippocampal synaptic plasticity and behavioral abnormalities related to spatial learning and memory. The molecular mechanisms leading to these impairments have not been identified. In this study, we focused on the G-protein-activated inwardly rectifying potassium channel 2 (GIRK2) gene that is highly expressed in the hippocampus region. We studied the expression pattern of GIRK subunits in Ts65Dn and found that GIRK2 was overexpressed in all analyzed Ts65Dn brain regions. Interestingly, elevated levels of GIRK2 protein in the Ts65Dn hippocampus and frontal cortex correlated with elevated levels of GIRK1 protein. This suggests that heteromeric GIRK1-GIRK2 channels are overexpressed in Ts65Dn hippocampus and frontal cortex, which could impair excitatory input and modulate spike frequency and synaptic kinetics in the affected regions. All GIRK2 splicing isoforms examined were expressed at higher levels in the Ts65Dn in comparison to the diploid hippocampus. The pattern of GIRK2 expression in the Ts65Dn mouse brain revealed by in situ hybridization and immunohistochemistry was similar to that previously reported in the rodent brain. However, in the Ts65Dn mouse a strong immunofluorescent staining of GIRK2 was detected in the lacunosum molecular layer of the CA3 area of the hippocampus. In addition, tyrosine hydroxylase containing dopaminergic neurons that coexpress GIRK2 were more numerous in the substantia nigra compacta and ventral tegmental area in the Ts65Dn compared to diploid controls. In summary, the regional localization and the increased brain levels coupled with known function of the GIRK channel may suggest an important contribution of GIRK2 containing channels to Ts65Dn and thus to DS neurophysiological phenotypes. PMID- 16374809 TI - Afferents to the orexin neurons of the rat brain. AB - Emotions, stress, hunger, and circadian rhythms all promote wakefulness and behavioral arousal. Little is known about the pathways mediating these influences, but the orexin-producing neurons of the hypothalamus may play an essential role. These cells heavily innervate many wake-promoting brain regions, and mice lacking the orexin neurons have narcolepsy and fail to rouse in response to hunger (Yamanaka et al. [2003] Neuron 38:701-713). To identify the afferents to the orexin neurons, we first injected a retrograde tracer into the orexin neuron field of rats. Retrogradely labeled neurons were abundant in the allocortex, claustrum, lateral septum, bed nucleus of the stria terminalis, and in many hypothalamic regions including the preoptic area, dorsomedial nucleus, lateral hypothalamus, and posterior hypothalamus. Retrograde labeling in the brainstem was generally more modest, but labeling was strong in the periaqueductal gray matter, dorsal raphe nucleus, and lateral parabrachial nucleus. Injection of an anterograde tracer confirmed that most of these regions directly innervate the orexin neurons, with some of the heaviest input coming from the lateral septum, preoptic area, and posterior hypothalamus. In addition, hypothalamic regions preferentially innervate orexin neurons in the medial and perifornical parts of the field, but most projections from the brainstem target the lateral part of the field. Inputs from the suprachiasmatic nucleus are mainly relayed via the subparaventricular zone and dorsomedial nucleus. These observations suggest that the orexin neurons may integrate a variety of interoceptive and homeostatic signals to increase behavioral arousal in response to hunger, stress, circadian signals, and autonomic challenges. PMID- 16374811 TI - Expression pattern of calcitonin gene-related peptide in the superior colliculus during postnatal development: demonstration of its intrinsic nature and possible roles. AB - Calcitonin gene-related peptide (CGRP) is a widespread neuropeptide with multiple central and peripheral targets. In an analysis on the expression of this peptide throughout the rat brain during postnatal development, we observed a discrepancy between results obtained by immunohistochemistry and by in situ hybridization. In the superior colliculus (SC), only the immunohistochemical signal could be detected (Terrado et al. [1997] Neuroscience 80:951-970). Here we focus our attention on this structure because the temporal pattern of CGRP immunoreactivity observed in the SC suggested the participation of this peptide in the postnatal maturation of the SC. In the present study, we describe in detail the postnatal development of collicular CGRP-immunoreactive structures and their spatiotemporal relationship with cholinergic modules and definitively demonstrate the local expression of CGRP in the SC. CGRP-immunopositive axons and neurons were distributed within the most ventral part of superficial strata and in the intermediate strata of the SC, showing a peak in staining intensity and density at the end of the first postnatal week. At P14, CGRPergic terminal fibers are arranged in small, clearly defined patches in a complementary manner with respect to the cholinergic modules, which start forming at this stage. By using Western blot and RT-PCR analyses, and by means of injections of antisense oligonucleotides, both the presence of CGRP peptide in the SC and the local expression of alpha-CGRP transcripts in collicular neurons were demonstrated. A possible role of CGRP is discussed in the context of postnatal modular compartmentalization of collicular afferents. PMID- 16374812 TI - Developmental period for N-methyl-D-aspartate (NMDA) receptor-dependent synapse elimination correlated with visuotopic map refinement. AB - During a short perinatal interval, N-methyl-D-aspartate receptor (NMDAR) function is essential to a process in which spontaneous retinal waves focus retinal axon arbors in the superficial layers of the rodent superior colliculus (sSC). Here we provide evidence that this NMDAR-dependent axonal refinement occurs through elimination of uncorrelated retinal synapses arising from disparate loci, rather than stabilization of topographically appropriate inputs. The density of synaptic release sites within fluorescently labeled retinal terminals was counted in double-labeling experiments using confocal microscopy and antibodies against synaptophysin or synapsin-1. Chronic NMDAR blockade from birth increased retinal axon synapse density at postnatal days (P) 6, 8, and 10, suggesting that NMDAR currents reduce synapse density during the refinement period. With assay at P14, after focal arborization has been established, the effect disappeared. Conversely, chronic NMDA treatment, known to induce functional synaptic depression in the sSC, decreased retinocollicular synapse density at P14, but not earlier, during the refinement period (P8). Thus during the development of retinocollicular topographic order, there is a period when NMDAR activity predominantly eliminates retinal axon synapses. We were able to extend this period by using retinal lesions to reduce synaptic density in a defined zone. Synapse density on intact retinocollicular axons sprouting into this zone was increased by NMDAR blockade, even when examined at P14. Thus, the period of NMDAR dependent synaptic destabilization is terminated by a factor related to the density and refinement of retinal arbors. PMID- 16374813 TI - Astrocytic and neuronal localization of the scaffold protein Na+/H+ exchanger regulatory factor 2 (NHERF-2) in mouse brain. AB - The Na+/H+ exchanger regulatory factor 2 (NHERF-2) is a scaffold protein that regulates cellular signaling by forming protein complexes. Several proteins known to interact with NHERF-2 are abundantly expressed in the central nervous system, but little is known about NHERF-2 localization in the brain. By using immunohistochemistry combined with light and electron microscopy, we found that many populations of astrocytes, as well as some populations of neurons, were immunopositive for NHERF-2 throughout the mouse brain. Quantitative analysis of the subcellular distribution of NHERF-2 immunostaining in four brain structures, cerebral cortex, hippocampus, striatum, and cerebellar cortex, showed that NHERF 2 was expressed mainly in astrocytic processes but was also sometimes observed in both pre- and postsynaptic neuronal elements. NHERF-2 immunostaining was associated mainly with the plasma membrane of neurons and astrocytes. However, NHERF-2 immunoreactivity was also observed in association with synaptic vesicles in putative glutamatergic axon terminals. The subcellular localization of NHERF-2 in brain is consistent with a role for NHERF-2 in forming complexes between cell surface and cytosolic proteins, and the preferential expression of NHERF-2 in astrocytes suggests that this scaffold protein may play an important role in astrocytic physiology. PMID- 16374814 TI - Immunohistochemical localization of calbindin-D28k and calretinin in the spinal cord of Xenopus laevis. AB - Immunohistochemical techniques were used to investigate the distribution and morphology of neurons containing the calcium-binding proteins calbindin-D28k (CB) and calretinin (CR) in the spinal cord of Xenopus laevis and determine the extent to which this organization is comparable to that of mammals. Most CB- and CR containing neurons were located in the superficial dorsal gray field, but with distinct topography. The lateral, ventrolateral, and ventromedial fields also possessed abundant neurons labeled for either CB or CR. Double immunohistofluorescence demonstrated that a subpopulation of dorsal root ganglion cells and neurons in the dorsal and ventrolateral fields contained CB and CR. By means of a similar technique, a cell population in the dorsal field was doubly labeled only for CB and nitric oxide synthase (NOS), whereas in the ventrolateral field colocalization of NOS with CB and CR was found. Choline acetyltransferase immunohistochemistry revealed that a subpopulation of ventral horn neurons, including motoneurons, colocalized CB and CR. The involvement of CB- and CR containing neurons in ascending spinal projections was demonstrated combining the retrograde transport of dextran amines and immunohistochemistry. Cells colocalizing the tracer and CB or CR were quite numerous, primarily in the dorsal and ventrolateral fields. Similar experiments demonstrated supraspinal projections from CB- and CR-containing cells in the brainstem and diencephalon. The distribution, projections, and colocalization with neurotransmitters of the neuronal systems containing CB and CR in Xenopus suggest that CB and CR are important neuromodulator substances with functions conserved in the spinal cord from amphibians through mammals. PMID- 16374815 TI - Calretinin immunoreactivity in the brain of the zebrafish, Danio rerio: distribution and comparison with some neuropeptides and neurotransmitter synthesizing enzymes. II. Midbrain, hindbrain, and rostral spinal cord. AB - The distribution of calretinin (CR) in the brainstem and rostral spinal cord of the adult zebrafish was studied by using immunocytochemical techniques. For analysis of some brainstem nuclei and regions, CR distribution was compared with that of complementary markers (choline acetyltransferase, glutamic acid decarboxylase, tyrosine hydroxylase, neuropeptide Y). The results reveal that CR is a marker of various neuronal populations distributed throughout the brainstem, including numerous cells in the optic tectum, torus semicircularis, secondary gustatory nucleus, reticular formation, somatomotor column, gustatory lobes, octavolateral area, and inferior olive, as well as of characteristic tracts of fibers and neuropil. These results indicate that CR may prove useful for characterizing a number of neuronal subpopulations in zebrafish. Comparison of the distribution of CR observed in the brainstem of zebrafish with that reported in an advanced teleost (the gray mullet) revealed a number of similarities, and also some interesting differences. Our results indicate that many brainstem neuronal populations have maintained the CR phenotype in widely divergent teleost lines, so CR studies may prove very useful for comparative analysis. PMID- 16374816 TI - Septal organ of Gruneberg is part of the olfactory system. AB - The olfactory system in rodents and many other mammals is classically divided into two anatomically separate, and morphologically distinct, sensory systems: the main olfactory system and the accessory olfactory system. We have now identified a novel third population of olfactory marker protein-expressing sensory neurons that is located in a discrete pocket of the rostral nasal septum, which we refer to as the septal organ of Gruneberg (SOG). Neurons in this region of the septum are located in the submucosa, in small grape-like clusters, rather than in a pseudostratified neuroepithelium, as seen in both the olfactory and vomeronasal neuroepithelia. Despite their unusual location, axons projecting from the SOG neurons fasciculate into several discrete bundles and terminate in a subset of main olfactory bulb glomeruli. These glomeruli most likely represent a subset of atypical glomeruli that are spatially restricted to the caudal main olfactory bulb. The unique rostral position of the SOG suggests that the SOG may be functionally specialized for the early detection of biologically relevant odorants. PMID- 16374817 TI - Clinical and pathological characterization of progressive aphasia. AB - OBJECTIVE: The clinical and neuropathological categorization of patients presenting with progressive aphasia is an area of controversy. This study aimed to characterize a large group of progressive aphasic patients from a single center (n = 38), first clinically by case note review, and then pathologically. METHODS: Hierarchical cluster analysis of the cases according to their clinical language deficits was used to establish an unbiased, data-driven classification. RESULTS: This analysis revealed two groups of cases corresponding to the syndromes of progressive nonfluent aphasia (n = 23) and semantic dementia (n = 15). Postmortem analysis showed a majority in both groups of pathologies from the spectrum of frontotemporal lobar degeneration: the most frequent were non Alzheimer's disease (AD) tauopathy in the nonfluent cases (10 of 23) and frontotemporal lobar degeneration with ubiquitin-positive, tau-negative inclusions in the fluent cases (8 of 15). Despite rigorous exclusion of cases with clinically significant memory deficits or other cognitive impairments, the pathology of AD was present in approximately one third of each group (overall 12 of 38), although often with an atypical neuroanatomical distribution. INTERPRETATION: Progressive aphasia is best seen as a composite of two conditions, on both clinical and pathological levels: progressive nonfluent aphasia and semantic dementia. PMID- 16374818 TI - Suppression of mitoxantrone cardiotoxicity in multiple sclerosis patients by dexrazoxane. AB - OBJECTIVE: To explore the potential of dexrazoxane to suppress subclinical cardiotoxicity in MS patients receiving mitoxantrone. METHODS: An open-label study was performed to evaluate possible subclinical cardiotoxicity in multiple sclerosis patients treated quarterly with mitoxantrone (48 mg/m(2) cumulative), with and without concomitant dexrazoxane, using blinded serial radionucleide ventriculography. RESULTS: No patient experienced symptoms of heart failure. Patients receiving dexrazoxane, which is cardioprotective for anthracyclines, exhibited a significantly lesser decline in left ventricular ejection fraction (mean change, -3.80% vs -8.55%, p < 0.001). INTERPRETATION: These results support a cardioprotective effect of dexrazoxane in mitoxantrone treated multiple sclerosis patients. PMID- 16374819 TI - An association between type 1 diabetes and idiopathic generalized epilepsy. AB - OBJECTIVE: Idiopathic generalized epilepsies (IGEs) account for approximately 30% of all patients with epilepsy. Both the IGEs and type 1 diabetes mellitus (T1D) represent serious worldwide problems, because of related medical and social management costs. Clinical experience suggested the two conditions were seen in individuals more frequently than might be expected by chance. METHODS: We compared the population prevalence of T1D in 15- to 30-year-olds to a cohort of 518 15- to 30-year-olds with IGE. RESULTS: We found a highly significant excess of T1D in our IGE cohort, with an odds ratio of 4.4 (95% confidence interval, 2.1 9.2). INTERPRETATION: Our results suggest that the prevalence of T1D is increased by a factor of four in young adults with IGE. To our knowledge, this is the first published association between the two conditions and expands the diseases known to be associated with T1D. PMID- 16374820 TI - Occupational magnetic field exposure and the risk of acoustic neuroma. AB - BACKGROUND: Acoustic neuroma is the intracranial tumor subtype showing the highest relative risk in relation to ionizing radiation but other environmental risk factors are largely unknown. This study was performed to investigate the effect of power frequency magnetic fields. METHOD: A total of 793 cases between 1987 and 1999 were identified through the Swedish cancer registry and 101,762 controls were randomly selected from the total population. Information about occupation was obtained from censuses and linked to gender specific job-exposure matrices based on actual measurements of 50 Hz magnetic field exposure. RESULT: We investigated time-weighted average, peak values, and rate of change of magnetic field exposure considering several different time windows in relation to cancer diagnosis. We found no increases in risks regardless of exposure metric or time window of exposure. CONCLUSION: This study is the largest ever on acoustic neuroma and the first study to evaluate this tumor subtype specifically in relation to extremely low frequency magnetic fields. The results do not support the hypothesis that 50 Hz magnetic fields increase the risk of acoustic neuroma. PMID- 16374822 TI - Substantia nigra tangles are related to gait impairment in older persons. AB - OBJECTIVE: Parkinsonian signs, especially gait impairment, are common and associated with morbidity and mortality in older persons. Our objective was to test the hypothesis that substantia nigra neurofibrillary tangles (NFTs) are related to parkinsonian signs in older persons with and without dementia. METHODS: We studied 86 deceased older Catholic clergy without idiopathic Parkinson's disease from the Religious Order Study, a longitudinal clinical pathological study. Mean age at death was 85.3 years. Signs of gait disturbance, bradykinesia, rigidity, and tremor were assessed proximate to death using a modified Unified Parkinson's Disease Rating Scale. Forty-micrometer paraffin embedded sections of substantia nigra were bleached before tau immunohistochemistry and the optical disector was used to count NFTs. We used multivariable linear regression to examine parkinsonian signs as a function of nigra NFTs, controlling for age, sex, education, and cortical NFTs. RESULTS: Substantia nigra NFTs were present in 67 of 86 persons (77.9%). After controlling for age, sex, education, and cortical NFTs, nigra NFTs were related to gait impairment (p < 0.001), but not bradykinesia, rigidity, or tremor. Results were not confounded by dementia, Braak score, neuroleptic medication, cerebral infarcts, or Lewy bodies. INTERPRETATION: NFTs in the substantia nigra are associated with gait impairment in older persons with and without dementia. PMID- 16374823 TI - Widespread decrease of nicotinic acetylcholine receptors in Parkinson's disease. AB - OBJECTIVE: Nicotinic acetylcholine receptors have close interactions with the dopaminergic system and play critical roles in cognitive function. The purpose of this study was to compare these receptors between living PD patients and healthy subjects. METHODS: Nicotinic acetylcholine receptors were imaged in 10 nondemented Parkinson's disease patients and 15 age-matched healthy subjects using a single-photon emission computed tomography ligand [(123)I]5-iodo-3-[2(S) 2-azetidinylmethoxy]pyridine. Using an arterial input function, we measured the total distribution volume (V; specific plus nondisplaceable), as well as the delivery (K(1)). RESULTS: Parkinson's disease showed a widespread significant decrease (approximately 10%) of V in both cortical and subcortical regions without a significant change in K(1). INTERPRETATION: These results indicate the importance of extending the study to demented patients. PMID- 16374824 TI - Migraine with aura is a risk factor for unprovoked seizures in children. AB - OBJECTIVE: Migraine is associated with epilepsy, but the time order and nature of the relationship are unclear. We conducted a population based case control study to clarify the time order to determine whether migraine is a risk factor for epilepsy. METHODS: Migraine symptoms were evaluated in a population-based case control study of all incident epilepsy in Icelandic children and in matched controls (next two same sex births in the country). RESULTS: Migraine was associated with a fourfold increased risk for developing epilepsy, an association explained by migraine with aura (odds ratio, 8.1; 95% confidence interval, 2.7 24.3). Migraine without aura did not increase risk for epilepsy. INTERPRETATION: Children with migraine with aura have a substantial increased risk to develop subsequent epilepsy. This finding is consistent with the hypothesis that migraine with aura and migraine without aura may be different disorders. PMID- 16374825 TI - Pregabalin-withdrawal encephalopathy and splenial edema. PMID- 16374826 TI - Sickle cell disease and electroencephalogram hyperventilation. PMID- 16374827 TI - Spatial distribution of hemorrhages in cerebral amyloid angiopathy. PMID- 16374828 TI - Neonatal porencephaly and adult stroke related to mutations in collagen IV A1. AB - OBJECTIVE: The objective of this study was to describe leukoencephalopathy, lacunar infarcts, microbleeds and macrobleeds in the context of a collagen IV A1 mutation. METHODS: We examined a family with autosomal dominant porencephaly, in whom a defect in collagen IV A1 was detected recently. The patients underwent neurological, ophthalmological, and cardiological examinations and magnetic resonance imaging of the brain. Electron microscopy of a skin biopsy was performed. Extensive laboratory screening was performed for thrombophilia and increased bleeding tendency. RESULTS: The porencephaly was symptomatic in the infantile period in two patients, whereas it led to only minor neurological dysfunction in their affected mother. However, she experienced development of recurrent strokes in her 40s. In addition to the porencephaly, all patients had a leukoencephalopathy, which was most severe in the mother. Her magnetic resonance imaging results also showed lacunar infarcts, macrobleeds and a multitude of microbleeds. No other risk factors for recurrent stroke were found. Electron microscopy showed interruptions of the basement membrane of skin capillaries and inhomogeneous thickening of the basement membrane with pools of basement membrane fragments. INTERPRETATION: Leukoencephalopathy, ischemic infarcts, microbleeds, and macrobleeds are indicative of an underlying microangiopathy, of which the best-known causes are hypertension, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy, and cerebral amyloid angiopathy. Mutations in collagen IV A1, a major component of the vascular basement membrane, appear to be another risk factor. PMID- 16374829 TI - Spastic paraplegia type 2 associated with axonal neuropathy and apparent PLP1 position effect. AB - OBJECTIVE: To report an association between spastic paraplegia type 2 with axonal peripheral neuropathy and apparent proteolipid protein gene (PLP1) silencing in a family. METHODS: Pulsed-field gel electrophoresis, custom array comparative genomic hybridization, and semi-quantitative multiplex polymerase chain reaction analyses were used to examine the PLP1 genomic region. RESULTS: Electrodiagnostic studies and a sural nerve biopsy showed features of a dystrophic axonal neuropathy. Molecular studies identified a small duplication downstream of PLP1. INTERPRETATION: We propose the duplication to result in PLP1 gene silencing by virtue of a position effect. Our observations suggest that genomic rearrangements that do not include PLP1 coding sequences should be considered as yet another potential mutational mechanism underlying PLP1-related dysmyelinating disorders. PMID- 16374830 TI - Lung cancer mortality among workers at a nuclear materials fabrication plant. AB - BACKGROUND: The Oak Ridge, Tennessee Y-12 plant has operated as a nuclear materials fabrication plant since the 1940s. Given the work environment, and prior findings that lung cancer mortality was elevated among white male Y-12 workers relative to US white males, we investigated whether lung cancer mortality was associated with occupational radiation exposures. METHODS: A cohort of 3,864 workers hired between 1947 and 1974 who had been monitored for internal radiation exposure was identified. Vital status was ascertained through 1990. RESULTS: Over the study period 111 lung cancer deaths were observed. Cumulative external radiation dose under a 5-year lag assumption was positively associated with lung cancer mortality (0.54% increase in lung cancer mortality per 10 mSv, se=0.16, likelihood ratio test (LRT)=5.84, 1 degree of freedom [df]); cumulative internal radiation dose exhibited a highly-imprecise negative association with lung cancer mortality. DISCUSSION: The positive association between external radiation dose and lung cancer mortality was primarily due to exposure occurring in the period 5 14 years after exposure (0.97% increase in lung cancer mortality rate per 10 mSv, se=0.28, LRT=6.35, 1 df). The association between external radiation dose and lung cancer mortality was negative for exposures occurring at ages<35 years and positive for exposures occurring at ages 35-50 and 50+years. CONCLUSIONS: There is evidence of a positive association between cumulative external radiation dose and lung cancer mortality in this population. However, a causal interpretation of this association is constrained by the uncertainties in external and internal radiation dose estimates, the lack of information about exposures to other lung carcinogens, and the limited statistical power of the study. PMID- 16374831 TI - Role of the Na+/Ca2+ exchanger in calcium homeostasis and human sperm motility regulation. AB - A number of cell functions, such as flagellar beating, swimming velocity, acrosome reaction, etc., are triggered by a Ca2+ influx across the cell membrane. For appropriate physiological functions, the motile human sperm maintains the intracellular free calcium concentration ([Ca2+]i) at a submicromolar level. The objective of this study was to determine the role of the Na+/Ca2+ exchanger (NCX) in the maintenance of [Ca2+]i in human spermatozoa. Spermatozoa maintained in extracellular medium containing>or=1 microM Ca2+ exhibited motility similar to that of the control. In addition to several calcium transport mechanisms described earlier, we provide evidence that the NCX plays a crucial role in the maintenance of [Ca2+]i. Three chemically unrelated inhibitors of the NCX (bepridil, DCB (3',4'-dichlorobenzamil hydrochloride), and KB-R7943) all blocked human sperm motility in a dose and incubation time dependent manner. The IC50 values for bepridil, DCB, and KB-R7943 were 16.2, 9.8, and 5.3 microM, respectively. The treatment with the above-mentioned blockers resulted in an elevated [Ca2+]i and a decreased [Na+]i. The store-operated calcium channel (SOCC) inhibitor SKF 96365 also blocked the sperm motility (IC50=2.44 microM). The presence of the NCX antigen in the human spermatozoa was proven by flow cytometry, confocal laser scanning microscopy, and immunoblotting techniques. Calcium homeostasis of human spermatozoa is maintained by several transport proteins among which the SOCC and the NCX may play a major role. PMID- 16374832 TI - Regulation of levels of actin threonine phosphorylation during life cycle of Physarum polycephalum. AB - Under various environmental stresses, the true slime mold Physarum polycephalum converts into dormant forms, such as microcysts, sclerotia, and spores, which can survive in adverse environments for a considerable period of time. In drought induced sclerotia, actin is threonine phosphorylated, which blocks its ability to polymerize into filaments. It is known that fragmin and actin-fragmin kinase (AFK) mediate this phosphorylation event. In this work, we demonstrate that high levels of actin threonine phosphorylation are also found in other dormant cells, including microcysts and spores. As the threonine phosphorylation of actin in microcysts and sclerotia were induced by drought stress but not by other stresses, we suggest that drought stress is essential for actin phosphorylation in both cell types. Although characteristic filamentous actin structures (dot- or rod-like structures) were observed in microcysts, sclerotia, and spores, actin phosphorylation was not required for the formation of these structures. Prior to the formation of both microcysts and sclerotia, AFK mRNA expression was activated transiently, whereas fragmin mRNA levels decreased. Our results suggest that drought stress and AFK might be involved in the threonine phosphorylation of actin. PMID- 16374833 TI - A novel method to identify gene-gene effects in nuclear families: the MDR-PDT. AB - It is now well recognized that gene-gene and gene-environment interactions are important in complex diseases, and statistical methods to detect interactions are becoming widespread. Traditional parametric approaches are limited in their ability to detect high-order interactions and handle sparse data, and standard stepwise procedures may miss interactions that occur in the absence of detectable main effects. To address these limitations, the multifactor dimensionality reduction (MDR) method [Ritchie et al., 2001: Am J Hum Genet 69:138-147] was developed. The MDR is well-suited for examining high-order interactions and detecting interactions without main effects. The MDR was originally designed to analyze balanced case-control data. The analysis can use family data, but requires a single matched pair be selected from each family. This may be a discordant sib pair, or may be constructed from triad data when parents are available. To take advantage of additional affected and unaffected siblings requires a test statistic that measures the association of genotype with disease in general nuclear families. We have developed a novel test, the MDR-PDT, by merging the MDR method with the genotype-Pedigree Disequilibrium Test (geno PDT)[Martin et al., 2003: Genet Epidemiol 25:203-213]. MDR-PDT allows identification of single-locus effects or joint effects of multiple loci in families of diverse structure. We present simulations to demonstrate the validity of the test and evaluate its power. To examine its applicability to real data, we applied the MDR-PDT to data from candidate genes for Alzheimer disease (AD) in a large family dataset. These results show the utility of the MDR-PDT for understanding the genetics of complex diseases. PMID- 16374834 TI - Apolipoprotein E and obstructive sleep apnea: evaluating whether a candidate gene explains a linkage peak. AB - Evidence from both linkage analyses and association-based analyses has implicated Apoliprotein E (ApoE) as a disease susceptibility locus for obstructive sleep apnea. To further assess the putative role of ApoE in sleep apnea, we performed genotyping, association, and linkage analyses in a cohort assembled to investigate the genetic epidemiology of sleep apnea. Among a subset of the Caucasian families, ten microsatellites, spanning 20 cM, were genotyped in a region near ApoE on chromosome 19 where previous suggestive linkage had been demonstrated using a 9.1-cM genome-wide scan. Haseman-Elston regression analysis, conducted with these fine mapping markers (n=196 sibling pairs, 56 families), showed evidence for linkage to marker AFM210yg9 (p=0.00034), which was increased over that observed with the original scan. ApoE genotyping also was performed on a larger set of data (n=1,211 from 271 families, ages 3-85 years) from the cohort with available DNA. To determine whether the ApoE genotype explains the linkage peak, we included the ApoE genotype as a covariate in regression models. Inclusion of ApoE E2 allele as a covariate reduced the regression coefficient by 18%, suggesting that ApoE does not substantively explain the linkage signal. Finally, we repeated an association-based analysis in the larger sample of 1,211 individuals, and observed a higher prevalence of sleep apnea among individuals with the ApoE E2 allele. Overall, the evidence suggests that there is a disease susceptibility locus for obstructive sleep apnea in the region of ApoE, but ApoE itself is unlikely to be the causative locus. PMID- 16374835 TI - Tag SNP selection for Finnish individuals based on the CEPH Utah HapMap database. AB - The pattern and nature of linkage disequilibrium in the human genome is being studied and catalogued as part of the International HapMap Project [:2003 Nature 426:789-796]. A key goal of the HapMap Project is to enable identification of tag single nucleotide polymorphisms (SNPs) that capture a substantial portion of common human genetic variability while requiring only a small fraction of SNPs to be genotyped [International HapMap Consortium, 2005: Nature 437:1299-1320]. In the current study, we examined the effectiveness of using the CEU HapMap database to select tag SNPs for a Finnish sample. We selected SNPs in a 17.9-Mb region of chromosome 14 based on pairwise linkage disequilibrium (r(2)) estimates from the HapMap CEU sample, and genotyped 956 of these SNPs in 1,425 Finnish individuals. An excess of SNPs showed significantly different allele frequencies between the HapMap CEU and the Finnish samples, consistent with population-specific differences. However, we observed strong correlations between the two samples for estimates of allele frequencies, r(2) values, and haplotype frequencies. Our results demonstrate that the HapMap CEU samples provide an adequate basis for tag SNP selection in Finnish individuals, without the need to create a map specifically for the Finnish population, and suggest that the four-population HapMap data will provide useful information for tag SNP selection beyond the specific populations from which they were sampled. PMID- 16374836 TI - Sickness behaviors in chronic cholestasis: an immune-mediated process? PMID- 16374837 TI - Crystal structure of alanyl-tRNA synthetase editing-domain homolog (PH0574) from a hyperthermophile, Pyrococcus horikoshii OT3 at 1.45 A resolution. PMID- 16374838 TI - NPC2 is expressed in human and murine liver and secreted into bile: potential implications for body cholesterol homeostasis. AB - The liver plays a critical role in the metabolism of lipoprotein cholesterol and in controlling its elimination through the bile. Niemann-Pick type C 2 (NPC2), a cholesterol-binding protein, is key for normal intracellular trafficking of lipoprotein cholesterol, allowing its exit from the endolysosomal pathway into the metabolically active pool of the cell. In addition, NPC2 is a secretory protein from astrocytes and epididymal cells. Although NPC2 mRNA is detected in the liver, plasma and biliary NPC2 protein levels and function have not been reported. This study demonstrates that NPC2 is present in murine and human plasma and bile. In addition, hepatic NPC2 protein expression was dramatically increased in NPC1-deficient mice but not regulated by cholesterol feeding or pharmacological modulation of various nuclear receptors involved in cholesterol and bile acid metabolism. Interestingly, biliary NPC2 levels were 3-fold increased in gallstone-susceptible C57BL6/J versus gallstone-resistant BALB/c mice. Furthermore, NPC2 was exclusively found in the cholesterol pro-nucleating ConA-binding fraction of human bile. In conclusion, NPC2 is secreted from the liver into bile and plasma, where it may have a functional role in cholesterol transport in normal and disease conditions. PMID- 16374840 TI - Troglitazone inhibits tumor growth in hepatocellular carcinoma in vitro and in vivo. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) has been implicated in the differentiation and growth inhibition of cancer cells. We examined the effects of PPARgamma activation by troglitazone on hepatocellular carcinoma (HCC) cell growth, proliferation, and apoptosis in vitro and in vivo. We also studied relationships between PPARgamma activation and cyclooxygenase-2 (COX-2) expression. Human HCC cell lines Huh7 and Hep3B were cultured in the presence or absence of troglitazone. Cell growth was determined via WST-1 assay, proliferation by cell cycle analysis and proliferating cell nuclear antigen (PCNA) Western blotting, and apoptosis by flow cytometry and TUNEL. Tumor growth after subcutaneous implantation of Huh7 cells in nude mice was monitored, and the effects of treatment with troglitazone were determined. In resected HCCs, PPARgamma expression was less compared with the histologically normal surrounding liver. In cultures of Hep3B and Huh7 cells, basal expression of PPARgamma was relatively low, but troglitazone caused dose-dependent induction of PPARgamma expression. Cell cycle analysis revealed a decreased proportion of cells in S phase, with arrest at G0/G1. Concomitant downregulation of PCNA and an increase in TUNEL staining, cells were consistent with decreased proliferation and induction of apoptosis by troglitazaone. Troglitazone-mediated PPARgamma activation also suppressed COX-2 expression and induced p27 in HCC cells. Administration of troglitazone to Huh7 tumor-bearing mice significantly reduced tumor growth and caused tumor regression. In conclusion, collectively, these results indicate that PPARgamma could be a regulator of cell survival and growth in HCC. PPARgamma therefore represents a putative molecular target for chemopreventive therapy or inhibition of liver cancer growth. PMID- 16374841 TI - Parenteral nutrition-associated liver disease and the role for isolated intestine and intestine/liver transplantation. AB - Parenteral nutrition-associated liver disease (PNALD) is the most devastating complication of long-term parenteral nutrition therapy. Because its progression is typically insidious and its long-term consequences are generally underappreciated, PNALD is often recognized too late, when liver injury is irreversible. When end-stage liver disease (ESLD) develops in these patients, most potential interventions are futile and transplantation of both an intestine and a liver becomes the only viable option, despite the relatively poor outcomes associated with this combined procedure. Although likely multifactorial in origin, the etiology of PNALD is poorly understood. Early clinical intervention with a combination of nutritional, medical, hormonal, and surgical therapies can be effective in preventing liver disease progression. If these interventions fail, intestinal transplantation should be performed expeditiously before development of ESLD mandates simultaneous inclusion of a liver graft as well. PMID- 16374842 TI - Crystal structure of a putative pyridoxine 5'-phosphate oxidase (Rv2607) from Mycobacterium tuberculosis. AB - The three-dimensional structure of Rv2607, a putative pyridoxine 5'-phosphate oxidase (PNPOx) from Mycobacterium tuberculosis, has been determined by X-ray crystallography to 2.5 A resolution. Rv2607 has a core domain similar to known PNPOx structures with a flavin mononucleotide (FMN) cofactor. Electron density for two FMN at the dimer interface is weak despite the bright yellow color of the protein solution and crystal. The shape and size of the putative binding pocket is markedly different from that of members of the PNPOx family, which may indicate some significant changes in the FMN binding mode of this protein relative to members of the family. PMID- 16374843 TI - Endoscopy critics vs. endoscopy enthusiasts for primary prophylaxis of variceal bleeding. PMID- 16374844 TI - Hematopoietic cells as hepatocyte stem cells: a critical review of the evidence. AB - The authors reviewed 77 published reports available before August 1, 2005 that examined the ability of hematopoietic cells to generate hepatocytes in the liver. A list of these publications and a synopsis of each are available on-line. We interpret the evidence provided by this data set to suggest that one or more types of hematopoietic cells may rarely acquire the hepatocyte phenotype in the liver (frequency < or =10(-4)), although the nature of the hematopoietic cells involved and the mechanisms responsible for acquisition of a hepatocyte phenotype are still controversial. Hematopoietic stem cells do not appear to be direct precursors of hepatocytes, which, instead, can be generated from cells of the macrophage-monocyte lineage. Fusion between hepatocytes and transplanted hematopoietic cells has been substantiated as a mechanism by which hepatocytes that carry a bone marrow tag are generated, but direct transdifferentiation of hematopoietic cells has not been demonstrated. In conclusion, hematopoietic cells contribute little to hepatocyte formation under either physiological or pathological conditions, although they may provide cytokines and growth factors that promote hepatocyte functions by paracrine mechanisms. Cells of the endodermal hepatocyte lineage are far more potent generators of hepatocytes than are hematopoietic cells. PMID- 16374846 TI - Systemic hemodynamics, vasoactive systems, and plasma volume in patients with severe Budd-Chiari syndrome. AB - Budd-Chiari syndrome (BCS) causes postsinusoidal portal hypertension, which leads to complications similar to those observed in cirrhosis. However, no studies have investigated whether patients with BCS develop the hyperdynamic circulatory syndrome present in patients with cirrhosis who have portal hypertension. We evaluated systemic and cardiopulmonary hemodynamics, plasma renin activity, aldosterone and norepinephrine levels, and plasma volume in patients with BCS admitted for complications of portal hypertension. BCS patients had mean systemic and cardiopulmonary pressures and cardiac indices that were within the normal range but were significantly different from those of a group of patients with cirrhosis matched by sex, body surface, and liver function (cardiac index 3.1 +/- 0.7 vs. 4.9 +/- 1.2 L.min(-1).m(-2); P < .001; systemic vascular resistance [SVR] index, 2,189 +/- 736 vs. 1,377 +/- 422 dyne.s.cm(-5).m(-2), P < .001). Despite normal systemic vascular resistance, BCS patients had activation of the neurohumoral vasoactive systems, as evidenced by increased plasma renin activity, aldosterone and norepinephrine levels (15.0 +/- 21.5 ng/mL . h, 76.7 +/- 106.8 ng/dL, 586 +/- 868 pg/mL; respectively) and plasma volume expansion. The analysis of individual BCS patients identified that 7 of the 21 patients actually had reduced SVR index. These patients had the greatest plasma volume expansion. A significant inverse correlation between plasma volume and SVR index was observed. In conclusion, patients with BCS had activation of vasoactive neurohumoral systems and expanded plasma volume. This outcome was observed even though most of these patients did not exhibit systemic vasodilation and cardiac output was not increased, in marked contrast with what is observed in patients with cirrhosis. PMID- 16374847 TI - Influence of beta-2 adrenergic receptor gene polymorphism on the hemodynamic response to propranolol in patients with cirrhosis. AB - The beta-2-adrenergic receptor (beta(2-)-AR) has several single-nucleotide polymorphisms. These influence the functional response to adrenergic stimulation; genotypes homozygous for Gly16-Glu27 or Gly16-Gln27 alleles (Gly16-Glu/Gln27 haplotypes) are associated with enhanced response, whereas genotypes homozygous for Arg16-Gln27 alleles (Arg16-Gln27) show a decreased response. We hypothesized that gene polymorphisms at the beta2-AR may influence the hemodynamic response to propranolol in patients with cirrhosis. The beta2-AR gene polymorphisms were determined by direct sequencing of the polymerase chain reaction (PCR) products in 48 patients with cirrhosis. All patients also had hepatic and systemic hemodynamic studies before and after propranolol administration. Prevalence of Gly16-Glu/Gln27 haplotypes was 29.1%, Arg16-Gln27 haplotype was 16.7%, and 54.2% were compound heterozygotes. Patients with cirrhosis with Gly16-Glu/Gln27 haplotypes had a greater decrease in heart rate, cardiac index, and hepatic blood flow after propranolol administration than those with Arg16-Gln27 haplotype. However, the HVPG response to propranolol was similar in both groups, whereas estimated hepatic sinusoidal resistance increased significantly in Gly16 Glu/Gln27 haplotypes but not in Arg16-Gln27 (+27.1 +/- 17.8% vs -17.9 +/- 13.9%, P = .042), suggesting that unopposed vasoconstrictive activity at the intrahepatic circulation hinders the fall in HVPG despite enhanced hemodynamic response to propranolol in Gly16-Glu/Gln27 haplotypes. In conclusion, beta2-AR gene polymorphisms influence the response to beta-blockade. However, HVPG reduction cannot be predicted from polymorphism analysis. Patients with the Gly16 Glu/Gln27 haplotypes may benefit from the association of hepatic vasodilators to propranolol therapy. PMID- 16374848 TI - Nrf2 is increased by CYP2E1 in rodent liver and HepG2 cells and protects against oxidative stress caused by CYP2E1. AB - Induction of CYP2E1 by ethanol is one pathway through which ethanol generates oxidative stress. Nrf2 is a transcription factor that regulates important antioxidant and phase II detoxification genes. Nrf2 induction by CYP2E1 and its importance in the adaptive response to increased oxidative stress caused by CYP2E1 was studied. Increases in Nrf2 protein and mRNA were observed in livers or hepatocytes of chronic alcohol-fed mice or rats and of pyrazole-treated rats or mice, conditions known to elevate CYP2E1. HepG2 cells expressing CYP2E1 (E47 cells) showed increased Nrf2 mRNA and protein expression compared with control HepG2 C34 cells. Nrf2 is activated in E47 cells as shown by an increase in nuclear Nrf2 levels and Nrf2-antioxidant-responsive element binding activity, and upregulation of Nrf2-regultated genes, glutamate cysteine ligase catalytic subunit (GCLC), and heme oxygenase 1 (HO-1). Increases in Nrf2 protein and mRNA are blocked by inhibitors of CYP2E1 activity and a reactive oxygen species (ROS) scavenger, N-acetylcysteine, which decrease ROS levels as well as Nrf2 mRNA induction. Upregulation of GCLC and HO-1 in E47 cells is dependent on Nrf2 and is prevented by siRNA-Nrf2. Blocking Nrf2 by siRNA-Nrf2 decreases glutathione and increases ROS and lipid peroxidation, resulting in decreased mitochondrial membrane potential and loss of cell viability of E47 cells but not C34 cells. These results suggest that Nrf2 is activated and that levels of protein and mRNA are increased when CYP2E1 is elevated. In conclusion, Nrf2 plays a key role in the adaptive response against increased oxidative stress caused by CYP2E1. PMID- 16374849 TI - TNF-alpha-secreting monocytes are recruited into the brain of cholestatic mice. AB - Signaling occurs between the liver and brain in cholestatic liver disease, giving rise to sickness behaviors such as fatigue. However, the signaling pathways involved are poorly defined. Circulating inflammatory mediator levels are increased in cholestasis, leading to speculation that they may be capable of activating circulating immune cells that subsequently could gain access to the brain. Indeed, we have identified that at day 10 after bile duct resection induced cholestasis, there is activation of circulating monocytes that express tumor necrosis factor alpha (TNF-alpha) in conjunction with increased expression of adhesion molecules by cerebral endothelium. Moreover, using intravital microscopy, we have identified markedly enhanced leukocytes rolling along cerebral endothelial cells, mediated by P-selectin, in bile duct-resected (BDR) but not control mice. In addition, we have identified increased infiltration of monocytes (but not lymphocytes) into the brains of BDR mice and found that these infiltrating monocytes produce TNF-alpha. Furthermore, infiltration of TNF-alpha secreting monocytes into the brains of cholestatic mice is associated with a broad activation of resident brain macrophages to produce TNF-alpha. In conclusion, cholestasis is associated with an activation of cerebral endothelium that recruits TNF-alpha-producing monocytes into the brain. We hypothesize that enhanced TNF-alpha release within the brain may contribute to the development of cholestasis-associated sickness behaviors, including fatigue. PMID- 16374850 TI - Dose and airflow dependence of benzyl alcohol disposition on skin. AB - The penetration of benzyl alcohol (BA) through split-thickness cadaver skin was measured in nonoccluded Franz cells placed in a fume hood. BA, dissolved in a small volume of ethanol and spiked with (14)C radiolabel, was applied to skin at nine doses ranging from 0.9 to 10600 microg/cm(2). The percentage of radioactivity penetrated after 24 h increased gradually with dose, ranging from 19.8 +/- 2.9% at the lowest dose to 29.2 +/- 3.0% at the highest. Less than 4% of the radioactivity was retained in the tissue at 24 h; the remainder was considered to be evaporated. These data and those from a previous study were analyzed in terms of a finite dose diffusion/evaporation model. The analysis showed that the increase in BA absorption with dose was consistent with a threefold increase in BA diffusivity in the stratum corneum, as its concentration increased from tracer levels to saturation. The variable diffusivity model was able to describe the combined observations from the two studies to within an rms error of 4.2% of dose. A method of estimating the diffusion model parameters independently of the experiment was found to yield good agreement with the experimentally-derived values at low and moderate doses. PMID- 16374851 TI - Alteration of the intravenous pharmacokinetics of a synthetic ozonide antimalarial in the presence of a modified cyclodextrin. AB - The pharmacokinetic profile and renal clearance of a novel synthetic ozonide antimalarial (1) was found to be significantly altered when intravenously administered to rats as a cyclodextrin-based formulation (0.1 M Captisol, a sulfobutylether beta-cyclodextrin derivative (SBE(7)-beta-CD)) compared to a cyclodextrin-free isotonic buffered glucose formulation. There was an 8.5-fold decrease in the steady-state blood volume of distribution, a 6.6-fold decrease in the mean residence time and a greater than 200-fold increase in renal clearance of 1 when administered in the cyclodextrin formulation. Analysis of the whole blood and plasma concentration profiles revealed an essentially constant blood to plasma ratio when 1 was administered in the cyclodextrin-free formulation, whereas this ratio changed as a function of time when administered in the presence of the cyclodextrin derivative. It is postulated that the observed differences were due to a very strong complexation interaction between 1 and the cyclodextrin, resulting in a slow dissociation of the complex in vivo, and altered distribution and excretion profiles. Preliminary studies using isothermal titration calorimetry (ITC) indicated that the association constant for the 1/Captisol complex was approximately two orders of magnitude higher than reported for typical drug/cyclodextrin complexes. PMID- 16374852 TI - Intraluminal drug and formulation behavior and integration in in vitro permeability estimation: a case study with amprenavir. AB - The purpose of this study was to assess the effect of biorelevant apical conditions on intestinal permeability estimation in the Caco-2 system for amprenavir, a poorly water-soluble substrate of the efflux carrier P-glycoprotein (P-gp). To establish biorelevant conditions, human intestinal fluids (HIF) were aspirated from the duodenum and jejunum in fasted subjects, before and during 4 h after the intake of a standard formulation of amprenavir (Agenerase). The HIF samples were characterized with respect to the concentrations of phospholipids, individual bile salts, amprenavir, and the excipient d-alpha-tocopheryl polyethyleneglycol 1000 succinate (TPGS); subsequently, the use of these samples in the Caco-2 system during permeability estimation for amprenavir was compared to standard conditions (amprenavir 10 microM dissolved in HBSS-based transport medium). The presence of the solubilizing excipient TPGS resulted in high intraluminal amprenavir concentrations (mM-range) and affected the permeability in a concentration-dependent way. At the observed intraluminal TPGS concentrations (mM-range), TPGS appeared to completely inhibit the interaction between amprenavir and P-gp, suggesting that the effect of P-gp on transepithelial transport of amprenavir in a clinical setting is probably negligible. This study illustrates the importance of the evaluation of intraluminal conditions after drug intake and their integration in permeability estimation in vitro. PMID- 16374853 TI - Nasobiliary drainage induces long-lasting remission in benign recurrent intrahepatic cholestasis. AB - Benign recurrent intrahepatic cholestasis (BRIC) is characterized by episodic cholestasis and pruritus without anatomical obstruction. Effective medical treatment is not available. We report complete and long-lasting disappearance of pruritus and normalization of serum bile salt concentrations in cholestatic BRIC patients within 24 hours after endoscopic nasobiliary drainage (NBD). Relative amounts of phospholipids and bile salts in bile collected during NBD appeared to be normal, but phospholipids other than phosphatidylcholine (especially sphingomyelin) were increased. In conclusion, we propose that temporary endoscopic nasobiliary drainage should be considered in cholestatic BRIC patients. PMID- 16374854 TI - LPS inhibits endothelin-1-induced endothelial NOS activation in hepatic sinusoidal cells through a negative feedback involving caveolin-1. AB - During endotoxemia, liver microcirculation disruption is characterized by a hypersensitivity to the constrictor effects of endothelin 1 (ET-1). The shift of ET-1-mediated effects toward vasoconstriction may result from depressed ET-1 mediated vasodilation through decreased ET-1-induced nitric oxide (NO) production. We have previously shown that lipopolysaccharide (LPS) pretreatment abrogates ET-1-induced endothelial nitric oxide synthase (eNOS) translocation, but its effects on eNOS activation are yet to be determined. Our aim was to assess the effects of LPS on ET-1-mediated eNOS activation in hepatic sinusoidal endothelial cells (SECs) and to investigate the molecular mechanisms involved. SECs were treated with LPS (100 ng/mL) for 6 hours followed by 30 minutes ET-1 (10 nmol/L) stimulation. LPS significantly inhibited ET-1-mediated eNOS activation. This inhibition was associated with upregulation of Caveolin-1 (CAV 1) and a shift in ET-1-mediated eNOS phosphorylation from an activation (Ser1177) to an inhibition (Thr495). LPS treatment has been shown to induce ET-1 expression and secretion from endothelial cells. We therefore investigated the role of endogenous ET-1 in the inhibition of ET-1 activation of eNOS after LPS. Antagonizing ET-1 effects and blocking its activation in LPS pretreated SECs decreased the LPS-induced overexpression of CAV-1 as well as the inhibition of ET 1-induced NOS activity. Furthermore, 6 hours of ET-1 treatment exerted the same effects on eNOS activity, phosphorylation, and CAV-1 expression as LPS treatment. In conclusion, LPS-induced suppression of ET-1-mediated eNOS activation is ET-1 dependent and suggest a pivotal role of CAV-1 in eNOS induction inhibition under stress. PMID- 16374855 TI - Limitation of combination therapy of interferon and ribavirin for older patients with chronic hepatitis C. AB - In contrast to the United States, Japanese patients with chronic hepatitis C currently treated with interferon are generally 10 to 15 years older. Older patients, however, tend to experience more frequent adverse events. This study was conducted to clarify the effect of patient age on the efficacy and safety of combination therapy. We consecutively enrolled 208 patients with naive chronic hepatitis C. Patients were classified into three groups according to age: younger than 50 years of age (n = 52); 50 to 59 years old (n = 83); and 60 years of age or older (n = 73). Interferon alpha-2b therapy was administered daily for 2 weeks, followed by 3 times per week for 22 weeks, while ribavirin was administered daily. Of the 208 study patients, discontinuation of therapy or dose reduction was required in 116 (56%) and was more frequent in older patient groups: 38%, 48%, and 77% for the < 50, 50-59, and > or = 60-year-old patient groups, respectively (P < .001). Multivariate analysis showed patient age to be independently associated with adherence to therapy. A sustained virological response was achieved in 77 (37%) patients, with genotype, viral load, and adherence to therapy associated with this achievement. A tendency toward a lower sustained virological response rate was seen in the older patients. In conclusion, patient age is an important factor contributing to the safety of combination therapy. Thus, treatment schedule should be modified, or other therapeutic modalities should be considered for older patients with chronic hepatitis C. PMID- 16374856 TI - Insulin resistance is associated with steatosis in nondiabetic patients with genotype 1 chronic hepatitis C. AB - Conflicting data exist regarding the relationship between hepatitis C virus genotype 1 and hepatic steatosis as well as the latter's role in the progression of fibrosis and treatment response. We assessed factors associated with hepatic steatosis in genotype 1 chronic hepatitis C and the impact of hepatic fat on fibrosis development and interferon responsiveness. Two hundred ninety-one non diabetic patients with genotype 1 chronic hepatitis C were examined for the presence of steatosis and its correlation with clinical, virological, and biochemical data, including insulin resistance (IR), evaluated by the homeostasis model assessment (HOMA) score. Steatosis was graded as mild (1%-20% of hepatocytes involved), moderate (21%-40% of hepatocytes involved), and severe (>40% of hepatocytes involved). Steatosis was mild in 110 of 291 (37.8%) and moderate/severe in 55 of 291 (18.9%) subjects. By logistic regression, moderate/severe steatosis was independently associated with the female sex (odds ratio [OR] 2.74; 95% CI 1.40-5.35), high gamma-glutamyltransferase levels (OR 1.52; 95% CI 1.22-1.91), and HOMA-score (OR 1.076; 95% CI 1.001-1.26). By logistic regression, moderate/severe steatosis (OR 2.78; 95% CI 1.21-6.4), and platelet counts (OR 0.97; 95% CI 0.96-0.98) were independent predictors of advanced fibrosis. Patients with moderate/severe steatosis had an OR of 0.52 (95% CI 0.30-0.90) for sustained virological response compared with patients with mild/absent steatosis. In conclusion, in nondiabetic European patients with genotype 1 hepatitis C at low risk for the metabolic syndrome, the prevalence of steatosis was nearly 60%. IR is a risk factor for moderate/severe steatosis, especially in men. Moderate/severe steatosis has clinical relevance, being associated with advanced fibrosis and hyporesponsiveness to antiviral therapy. PMID- 16374857 TI - Development and validation of two models for early prediction of response to therapy in genotype 1 chronic hepatitis C. AB - Early prediction of response to therapy in genotype 1 chronic hepatitis C is difficult. Two predictive models, a pretreatment scoring model (PreT-SM) and a fourth week of therapy scoring model (4w-SM) were constructed in a cohort of 104 patients from a single center (estimation cohort) and validated in a cohort of 141 patients from four independent centers (validation cohort). Individual scores were calculated using variables independently associated with sustained virological response (SVR). Baseline viral load, aspartate aminotransferase/alanine aminotransferase ratio, serum cholesterol, and a numerical score for noninvasive estimation of liver fibrosis were included in the PreT-SM; HCV RNA clearance and PreT-SM scores were included in the 4w-SM. Receiver operating characteristic analysis revealed the area under the curve in the estimation cohort and in the validation cohort to be, respectively, 0.856 and 0.847 for the PreT-SM and 0.908 and 0.907 for the 4w-SM. Low scores were associated with SVR, high scores with non-SVR. The best cutoff scores from the PreT-SM (7 and 9.70) identified, respectively, 36% of patients with SVR and 41% of those with non-SVR from the validation cohort, with high accuracy (> or =90% positive predictive value [PPV] and specificity). Similarly, cutoff scores of 3.20 and 5.60 from the 4w-SM identified, respectively, 71% of patients with SVR and 53% of those with non-SVR from the same cohort with high accuracy (PPV and specificity >92%). In conclusion, these models predicted response to therapy before or after 4 weeks of treatment in approximately 60% of genotype 1 patients and may be valuable for the management of this condition. PMID- 16374858 TI - JNK1 but not JNK2 promotes the development of steatohepatitis in mice. AB - Nonalcoholic fatty liver disease (NAFLD) is characterized by hepatic steatosis and varying degrees of necroinflammation. Although chronic oxidative stress, inflammatory cytokines, and insulin resistance have been implicated in the pathogenesis of NAFLD, the mechanisms that underlie the initiation and progression of this disease remain unknown. c-Jun N-terminal kinase (JNK) is activated by oxidants and cytokines and regulates hepatocellular injury and insulin resistance, suggesting that this kinase may mediate the development of steatohepatitis. The presence and function of JNK activation were therefore examined in the murine methionine- and choline-deficient (MCD) diet model of steatohepatitis. Activation of hepatic JNK, c-Jun, and AP-1 signaling occurred in parallel with the development of steatohepatitis in MCD diet-fed mice. Investigations in jnk1 and jnk2 knockout mice demonstrated that jnk1, but not jnk2, was critical for MCD diet-induced JNK activation. JNK promoted the development of steatohepatitis as MCD diet-fed jnk1 null mice had significantly reduced levels of hepatic triglyceride accumulation, inflammation, lipid peroxidation, liver injury, and apoptosis compared with wild-type and jnk2 -/- mice. Ablation of jnk1 led to an increase in serum adiponectin but had no effect on serum levels of tumor necrosis factor-alpha. In conclusion, JNK1 is responsible for JNK activation that promotes the development of steatohepatitis in the MCD diet model. These findings also provide additional support for the critical mechanistic involvement of JNK1 overactivation in conditions associated with insulin resistance and the metabolic syndrome. PMID- 16374861 TI - BetaKlotho: a new kid on the bile acid biosynthesis block. PMID- 16374859 TI - Exendin-4, a glucagon-like protein-1 (GLP-1) receptor agonist, reverses hepatic steatosis in ob/ob mice. AB - Nonalcoholic fatty liver disease (NAFLD) represents a burgeoning problem in hepatology, and is associated with insulin resistance. Exendin-4 is a peptide agonist of the glucagon-like peptide (GLP) receptor that promotes insulin secretion. The aim of this study was to determine whether administration of Exendin-4 would reverse hepatic steatosis in ob/ob mice. Ob/ob mice, or their lean littermates, were treated with Exendin-4 [10 microg/kg or 20 microg/kg] for 60 days. Serum was collected for measurement of insulin, adiponectin, fasting glucose, lipids, and aminotransferase concentrations. Liver tissue was procured for histological examination, real-time RT-PCR analysis and assay for oxidative stress. Rat hepatocytes were isolated and treated with GLP-1. Ob/ob mice sustained a reduction in the net weight gained during Exendin-4 treatment. Serum glucose and hepatic steatosis was significantly reduced in Exendin-4 treated ob/ob mice. Exendin-4 improved insulin sensitivity in ob/ob mice, as calculated by the homeostasis model assessment. The measurement of thiobarbituric reactive substances as a marker of oxidative stress was significantly reduced in ob/ob treated mice with Exendin-4. Finally, GLP-1-treated hepatocytes resulted in a significant increase in cAMP production as well as reduction in mRNA expression of stearoyl-CoA desaturase 1 and genes associated with fatty acid synthesis; the converse was true for genes associated with fatty acid oxidation. In conclusion, Exendin-4 appears to effectively reverse hepatic steatosis in ob/ob mice by improving insulin sensitivity. Our data suggest that GLP-1 proteins in liver have a novel direct effect on hepatocyte fat metabolism. PMID- 16374862 TI - Hepatocellular carcinoma in a 12-year-old child with PiZZ alpha1-antitrypsin deficiency. PMID- 16374863 TI - What is really ongoing during occult HBV reactivation? PMID- 16374864 TI - Is banding ligation for primary prevention of variceal bleeding as effective as beta-blockers, and is it safe? PMID- 16374865 TI - Outside St. Jorgen: leprosy in the medieval Danish city of Odense. AB - Leprosy was a common and dreaded disease in the Danish Middle Ages (AD 1050 1536). Starting in the second half of the 13th century, leprosaria were established in many Danish towns and cities. In the city of Odense (on the island of Funen, Denmark), the cemetery of the leprosarium was totally excavated, and four nonleprosarium medieval and early modern cemeteries have been partly excavated. This paper explores the frequency of leprosy in the nonleprosarium cemeteries in Odense, and looks for evidence of selective exclusion from the ordinary population. The analyses are based on 733 skeletons from four cemeteries in Odense: the Gray Friars monastery, St. Albani parish church, St. Knuds cathedral, and Black Friars monastery. Seven lesions are scored and, based on known epidemiological properties (i.e., specificity and sensitivity) of these lesions, scores were transformed to statistics characterizing an individual's risk of having suffered from leprosy. This statistical approach remains of primary theoretical value, pending confirmation by independent research groups at other sites. Prevalence of the skeletal manifestation of leprosy at death varied between 0-17% among the different cemeteries in Odense. The highest prevalence was seen in cemeteries with many burials before AD 1400. It is estimated that before AD 1400, between 14-17% of those buried in the nonleprosarium cemeteries suffered from leprosy. In all nonleprosarium cemeteries, there was evidence for selective exclusion of people with facial leprosy lesions. For a short period just up to AD 1300, the cemetery of the Odense leprosarium had, on average, more than 20 yearly burials. The establishment of the leprosarium was followed within a relatively short period by a dramatic decline in the number of sufferers of leprosy in the nonleprosarium cemeteries. The number of yearly burials in the leprosarium cemetery also declined rapidly during the 14th century. The present analyses do not permit conclusions about the reasons for this decline in leprosy prevalence. PMID- 16374866 TI - Skin and hair pigmentation variation in Island Melanesia. AB - Skin and hair pigmentation are two of the most easily visible examples of human phenotypic variation. Selection-based explanations for pigmentation variation in humans have focused on the relationship between melanin and ultraviolet radiation, which is largely dependent on latitude. In this study, skin and hair pigmentation were measured as the melanin (M) index, using narrow-band reflectance spectroscopy for 1,135 individuals from Island Melanesia. Overall, the results show remarkable pigmentation variation, given the small geographic region surveyed. This variation is discussed in terms of differences between males and females, among islands, and among neighborhoods within those islands. The relationship of pigmentation to age, latitude, and longitude is also examined. We found that male skin pigmentation was significantly darker than females in 5 of 6 islands examined. Hair pigmentation showed a negative, but weak, correlation with age, while skin pigmentation showed a positive, but also weak, correlation with age. Skin and hair pigmentation varied significantly between islands as well as between neighborhoods within those islands. Bougainvilleans showed significantly darker skin than individuals from any other island considered, and are darker than a previously described African-American population. These findings are discussed in relation to prevailing hypotheses about the role of natural selection in shaping pigmentation variation in the human species, as well as the role of demographic processes such as admixture and drift in Island Melanesia. PMID- 16374867 TI - Mechanistic link between the anti-HCV effect of interferon gamma and control of viral replication by a Ras-MAPK signaling cascade. AB - Interferon-gamma (IFN-gamma) exerts potent antiviral activity in the hepatitis C virus (HCV) replicon systems. However, the mechanisms underlying the direct antiviral effect have not been determined. We found that the type II transcriptional response to IFN-gamma could be suppressed by inhibition of MEK1/2 kinase activity by MEK1/2 inhibitor U0126 in the hepatoma cell line Huh-7. Using a bicistronic HCV replicon system expressing a luciferase reporter gene in Huh-7 cells (RLuc-replicon), we showed that inhibition of MEK1/2 kinase activity is sufficient to counteract the antiviral activity of IFN-gamma. Expression of a constitutive active form of Ras inhibited the luciferase activity of RLuc replicon, whereas a dominant-negative mutant of Ras enhanced the reporter activity, indicating that the Ras-MAPK pathway has a role in limiting replication of the viral RNA. Consistent with the involvement of the Ras-MAPK pathway, treatment with epidermal growth factor suppressed HCV protein expression in the RLuc-replicon cells, an effect that could be abolished by U0126. Inhibition of MEK1/2 kinase activity correlated with reduced phosphorylation of the HCV NS5A protein and enhanced RLuc-replicon luciferase reporter activity, in line with recent reports that phosphorylation of NS5A negatively modulates HCV RNA replication. Finally, genetic deletion analysis in yeast supported the role of a MEK-like kinase(s) in the regulation of NS5A phosphorylation. In conclusion, the direct anti-HCV effect of IFN-gamma in cell culture is, at least in part, mediated through the Ras-MAPK signaling pathway, which possibly involves a direct or indirect modulation of NS5A protein phosphorylation. PMID- 16374868 TI - Brain metabolism of 13N-ammonia during acute hepatic encephalopathy in cirrhosis measured by positron emission tomography. AB - Animal studies and results from 13N-ammonia positron emission tomography (PET) in patients with cirrhosis and minimal hepatic encephalopathy suggest that a disturbed brain ammonia metabolism plays a pivotal role in the pathogenesis of hepatic encephalopathy (HE). We studied brain ammonia kinetics in 8 patients with cirrhosis with an acute episode of clinically overt HE (I-IV), 7 patients with cirrhosis without HE, and 5 healthy subjects, using contemporary dynamic 13N ammonia PET. Time courses were obtained of 13N-concentrations in cerebral cortex, basal ganglia, and cerebellum (PET-scans) as well as arterial 13N-ammonia, 13N urea, and 13N-glutamine concentrations (blood samples) after 13N-ammonia injection. Regional 13N-ammonia kinetics was calculated by non-linear fitting of a physiological model of brain ammonia metabolism to the data. Mean permeability surface area product of 13N-ammonia transfer across blood-brain barrier in cortex, PS(BBB), was 0.21 mL blood/min/mL tissue in patients with HE, 0.31 in patients without HE, and 0.34 in healthy controls; similar differences were seen in basal ganglia and cerebellum. Metabolic trapping of blood 13N-ammonia in the brain showed neither regional, nor patient group differences. Mean net metabolic flux of ammonia from blood into intracellular glutamine in the cortex was 13.4 micromol/min/L tissue in patients with cirrhosis with HE, 7.4 in patients without HE, and 2.6 in healthy controls, significantly correlated to blood ammonia. In conclusion, increased cerebral trapping of ammonia in patients with cirrhosis with acute HE was primarily attributable to increased blood ammonia and to a minor extent to changed ammonia kinetics in the brain. PMID- 16374869 TI - Carbopol-mediated paracellular transport enhancement in Calu-3 cell layers. AB - The interaction of Carbopol polymers with mucus producing Calu-3 human bronchial epithelial cells was evaluated to test for potential paracellular transport enhancement. Using desmopressin (1-deamino-8-arginine-vasopressin, DDAVP) as the model peptide, apical treatment with Carbopol polymer gel formulations resulted in molecular size-dependent permeability enhancement with a concomitant drop in the transepithelial electrical resistance (TEER). Permeability enhancement of DDAVP was dependent on the formulation vehicle composition and polymer concentration, was noncytotoxic, and completely reversible. Carbopol 971P displayed the greatest permeability enhancement across Calu-3 cells compared to other more viscous Carbopol polymers 934P and 974P, and other mucoadhesive cellulosic polymers. The greatest enhancement was observed when C971P formulation was prepared in water at a concentration of 0.25% w/v. Enhancement was confirmed in rabbit dosed with intranasal fluorescent dextran 4400. The C(max) and absorption rate each increased by 48% in C971P formulations compared to control, while the relative exposure increased 30%. In conclusion, Carbopol polymers are potentially useful excipients to enhance intranasal peptide absorption. We hypothesize that the permeation enhancement is related to the chelation of extracellular or tight-junctional Ca(2+) by charged polymer carboxylate groups that leads to temporary disruption of tight-junctions, thereby facilitating paracellular transport. PMID- 16374871 TI - Untangling influences of hydrophobicity on protein sequences and structures. AB - We perform a statistical analysis of solvent accessibility and hydrophobicity profiles of a representative set of proteins. The joint probability distribution is well fitted to a multivariable Gaussian, which takes a relatively simple form when expressed in terms of the Fourier transforms of the profiles. This allows us to quantify the asymmetric manner by which these profiles influence each other. For example, the alpha-helix periodicity in sequence hydrophobicity is dictated by the solvent accessibility of structures, and not vice versa, possibly indicating the faster evolution of sequences compared to structures. The decorrelated hydrophobicity and solvent accessibility profiles show distinct behaviors at long periods, where sequence hydrophobicity fluctuates less, while solvent accessibility fluctuates more than average. The correlations between the two profiles can be interpreted as the Boltzmann weight of the solvation energy at room temperature, consistent with earlier observations. PMID- 16374872 TI - The role of residue stability in transient protein-protein interactions involved in enzymatic phosphate hydrolysis. A computational study. AB - Finding why protein-protein interactions (PPIs) are so specific can provide a valuable tool in a variety of fields. Statistical surveys of so-called transient complexes (like those relevant for signal transduction mechanisms) have shown a tendency of polar residues to participate in the interaction region. Following this scheme, residues in the unbound partners have to compete between interacting with water or interacting with other residues of the protein. On the other hand, several works have shown that the notion of active site electrostatic preorganization can be used to interpret the high efficiency in enzyme reactions. This preorganization can be related to the instability of the residues important for catalysis. In some enzymes, in addition, conformational changes upon binding to other proteins lead to an increase in the activity of the enzymatic partner. In this article the linear response approximation version of the semimacroscopic protein dipoles Langevin dipoles (PDLD/S-LRA) model is used to evaluate the stability of several residues in two phosphate hydrolysis enzymes upon complexation with their activating partners. In particular, the residues relevant for PPI and for phosphate hydrolysis in the CDK2/Cyclin A and Ras/GAP complexes are analyzed. We find that the evaluation of the stability of residues in these systems can be used to identify not only active site regions but it can also be used as a guide to locate "hot spots" for PPIs. We also show that conformational changes play a major role in positioning interfacing residues in a proper "energetic" orientation, ready to interact with the residues in the partner protein surface. Thus, we extend the preorganization theory to PPIs, extrapolating the results we obtained from the above-mentioned complexes to a more general case. We conclude that the correlation between stability of a residue in the surface and the likelihood that it participates in the interaction can be a general fact for transient PPIs. PMID- 16374873 TI - Hematopoietic mobilization in mice increases the presence of bone marrow-derived hepatocytes via in vivo cell fusion. AB - The mechanisms for in vivo production of bone marrow-derived hepatocytes (BMDHs) remain largely unclear. We investigated whether granulocyte colony-stimulating factor (G-CSF)-mediated mobilization of hematopoietic cells increases the phenomenon. Recurrent liver injury in mice expressing green fluorescent protein (EGFP) in all hematopoietic-derived cells was produced by 3 months of carbon tetrachloride (CCL4) injections. Histologically, there were necrotic foci with histiocyte-rich infiltrates, but little oval cell proliferation. Subsequently, some animals were mobilized with G-CSF for 1, 2, or 3 weeks. Animals were sacrificed 1 month after growth factor treatment. BMDH percentages were lower than previously reported, though G-CSF mobilization significantly augmented BMDH production in injured livers. BMDHs originating from in vivo fusion were evaluated by transplanting female EGFP+ cells into male mice. Binucleated, EGFP+ hepatocytes with one Y chromosome, indicating fusion, were identified. In conclusion, (1) mobilization of hematopoietic cells increases BMDH production and (2) as with the FAH-null model, the first model demonstrating hematopoietic/hepatocyte fusion, recurring CCl4-induced injury has macrophage rich infiltrates, a blunted oval cell response, and a predominantly in vivo fusion process for circulating cell engraftment into the liver. These findings open the possibility of using hematopoietic growth factors to treat nonhematopoietic degenerative diseases. PMID- 16374874 TI - Biorelevant dissolution media: aggregation of amphiphiles and solubility of estradiol. AB - Biorelevant dissolution media containing bile salt and lecithin at concentrations appropriate for fed and fasted state are useful when testing oral solid formulations of poorly water-soluble drugs. Dilution of amphiphile solutions affects the aggregation state of the amphiphiles because bile salt is partitioned between the aqueous phase and the aggregates. The aim of the investigation was to study the effect of dilution on the size distribution of aggregates and its effect on the solubilization capacity. Clear buffered solutions of four intestinal amphiphiles (sodium glycocholate, lecithin, monoolein, and oleic acid) and a combination of these were prepared at high bile salt concentration. Micelles in the glycocholate solutions decreased in size when diluted. The addition of insoluble amphiphiles led to bigger micelles with no clear correlation between size of the micelles and amphiphile concentration. Dilution of the two- and four component media caused enlargement of the mixed micelles and formation of vesicles. The solubility of estradiol in the buffer solution was increased with addition of the amphiphiles. A good correlation (R(2) = 0.987) was found between estradiol solubility and mass concentration of the amphiphiles. The results demonstrate that, in the case of estradiol, the concentration of amphiphiles rather than the aggregation state determines the solubilization capacity of the medium. PMID- 16374875 TI - Cardiac CINE MR imaging with a 32-channel cardiac coil and parallel imaging: impact of acceleration factors on image quality and volumetric accuracy. AB - PURPOSE: To assess the impact of parallel imaging algorithms on image quality and volumetric accuracy of CINE magnetic resonance imaging (MRI) with high temporal and spatial resolution using a new 32-channel dedicated cardiac phased array coil. MATERIALS AND METHODS: Fourteen individuals underwent steady-state free precession (SSFP) CINE MRI using a 32-element phased-array coil and parallel imaging acceleration using spatiotemporal sensitivity encoding (TSENSE). Acquisition acceleration ranged from R = 2 to 7. In conjunction with data extracted from phantom measurements, contrast-to-noise ratio (CNR) performance was evaluated for each acceleration factor and subjective image quality was evaluated by two independent readers. In addition, volumetric assessment was performed for each acceleration factor based on a single breath-hold multi-slice data acquisition. Results were compared to nonTSENSE measurements. RESULTS: CNR for non-accelerated CINE (R = 1) was 45.7 +/- 12.8 and showed a constant decrease with increase in acceleration of 51% at R = 4 and 86% at R = 7. CNR losses accompanied reductions in subjective image quality. Volumetric evaluation was accurate for R 0.05). For women over 40, rates of undergoing mammography screening were similar (p>0.05), but regular attendance was reported more by women with a family history (p<0.05). More women under 40 with a family history of breast cancer attended mammography (p<0.05), but only about 14% had ever undertaken mammography screening and 27% had ever undertaken CBE. More than 50% of the women had performed BSE, while significantly more women with a family history reported its over-performance (p<0.01). Women with a family history reported higher perceived susceptibility (p<0.01), higher cancer worries (p<0.05), and fewer barriers to mammography (p<0.05). According to logistic regression analysis, higher odds of EDP were significantly related to perceiving fewer barriers and having higher cancer worries. A positive family history was related to higher odds of women undergoing mammography. Perceived susceptibility was significantly related to higher odds of BSE only. Over-performance of BSE was significantly related to having a positive family history, higher susceptibility, and higher cancer worries. CONCLUSIONS: (1) A high rate of women did not undergo CBE or mammography screening. Women under 40 with a family history of breast cancer who have never undergone CBE or mammography merit special attention. (2) The change in guidelines on BSE necessitates further study of its over-performance in relation to cancer worries. (3) Interventions are needed to promote attendance for CBE and mammography in younger women with a positive family history. PMID- 16374895 TI - Homocysteine metabolism in families from southern Italy with neural tube defects: role of genetic and nutritional determinants. AB - OBJECTIVE: To evaluate the role of different polymorphic gene variants involved in homocysteine metabolism and plasma levels of homocysteine, folate and vitamin B12 in families from southern Italy with neural tube defects (NTDs). METHODS: Eighteen fathers, 15 NTD children and 60 women who had conceived NTD foetuses were investigated. A group of 100 adults and 43 apparently healthy children was used as control. At the time of blood draw, none were taking vitamin pills or nutritional supplements. RESULTS: Among controls, 79 (55.2%) were heterozygous for C677T MTHFR variant and 26 (18.2%) were TT homozygous. Among the cases, 35 (61.4%) out of 57 mothers and 7 (38.9%) out of 18 fathers carried the T allele; 12 (21.1%) mothers and 2 (11.1%) fathers had the TT genotype. Four (26.7%) out of 15 probands were TT homozygous and 11 (73.3%) were heterozygous (Fisher exact test p = 0.025). No significant difference between groups was observed for the 1298C MTHFR variant and CBS haplotypes. Median homocysteine in NTD children was significantly higher (10.0 micromol/L) than that of controls (median 4.5 micromol/L, Mann-Whitney p < 0.05). Folate and B12 were not different among groups. CONCLUSIONS: The T677 MTHFR allele is significantly associated with the occurrence of NTDs; no significant association has been observed with other genetic determinants analysed. Homocysteine levels in children with NTDs are significantly higher than those of the paediatric population from the same geographical area. PMID- 16374896 TI - Huge duplication cyst of small intestine: ultrasonographic features and prenatal aspiration. PMID- 16374897 TI - Quantification of fetal nucleated cells in maternal blood of pregnant women with a male trisomy 21 fetus using molecular cytogenetic techniques. AB - BACKGROUND: Prenatal diagnosis of trisomy 21 is based on fetal karyotyping generally obtained using invasive methods. During pregnancy, the circulating fetal cells in maternal blood constitute a potential source for development of a noninvasive prenatal diagnosis. The objective of this study was the identification and quantification of all fetal nucleated cells per unit volume of peripheral blood of pregnant women carrying male fetuses with trisomy 21 using molecular cytogenetic techniques. METHODS: Peripheral blood samples were obtained from 16 women carrying male fetuses with trisomy 21. We used a simple and rapid method of harvesting blood without recourse to any enrichment procedures or cell separation techniques. To evaluate the potential of this method, 16 specimens were analyzed by molecular cytogenetic techniques such as fluorescence in situ hybridization (FISH) and primed in situ labeling (PRINS) using specific probes to chromosomes X, Y and 21. RESULTS: The number of fetal cells varied between 6 and 32 per mL of maternal blood. This number is 3-5 times higher than that from normal pregnancies. CONCLUSIONS: Our current results are in agreement with the results previously reported by other groups showing that the number of fetal cells in maternal blood in trisomic 21 pregnancies is higher than in normal pregnancies. This high number of fetal cells is regarded as an advantage for the development of a noninvasive prenatal diagnostic test. PMID- 16374898 TI - Diagnosis, surveillance, and treatment of the anemic fetus using middle cerebral artery peak systolic velocity measurement. AB - The in utero course of the anemic fetus has improved dramatically, owing to early diagnosis and cordocentesis transfusion. In utero invasive procedures such as amnio- and cordocentesis have become important modalities in the evaluation and treatment of anemic fetuses. However, they carry risks for both the mother and fetus. A valid and sensitive noninvasive means of following the anemic fetus is the evaluation of changes in the middle cerebral artery peak systolic flow velocity (MCA-PSV). This is a sensitive tool for both the evaluation of fetal anemia and response to treatment. Intracerebral vessels respond earliest to the fetal anemic state, and are readily accessible for ultrasound examination. We describe the methodology and evolving clinical applications of MCA-PSV measurement in the fetus, through an overview of the literature describing the development and application of MCA-PSV measurement in fetuses at risk of fetal anemia of various immune and nonimmune etiologies, illustrated by index cases from our center. MCA-PSV measurement is essential in the diagnosis, evaluation, and management of cases of fetal anemia. The use of this modality lessens the need for invasive procedures. The method is readily accessible and should be integrated into the repertoire of all obstetric ultrasound centers. PMID- 16374899 TI - Validity of asthma diagnoses recorded in the Medical Services database of Quebec. AB - The goal of this study was to evaluate the validity of asthma diagnoses recorded in the Medical Services (physician billing) database of the Canadian province of Quebec. The predictive positive value (PPV) and predictive negative value (PNV) of two operational definitions of asthma based on diagnoses recorded in the database were evaluated. Patients 16-80 years old treated by a respiratory or a family physician in 2002 were selected from the database. The diagnosis derived from the Medical Services database was compared to the diagnosis written in the patient's medical chart. The PPV and PNV of the first operational definition based on one asthma diagnosis or more recorded in the database over a 1-year period were found to be 0.75 and 0.96 for respiratory physicians and 0.67 and 0.99 for family physicians, for patients 16-44 years old. The PPV increased to 0.78 for family physicians and to 0.77 for respiratory physicians when the second operational definition based on two diagnoses of asthma or more was used. Results tended to be lower for 45-80 years old patients. We conclude that diagnoses recorded in the Medical Services database of Quebec are valid to identify patients with asthma. PMID- 16374900 TI - Prenatal HLA-typing in beta-thalassemia before the collection of sibling cord blood. PMID- 16374901 TI - Fetal obstructive uropathy: is urine sampling useful for prenatal counselling? AB - OBJECTIVES: To evaluate whether fetal urinary sodium and chloride provide clinically useful information in addition to ultrasound in bilateral obstructive uropathy. METHODS: Sonographic features and urinary concentrations of sodium and chloride were evaluated in fetuses with bilateral obstructive uropathy. After a minimum of 12 months of postnatal follow-up, cases that developed increased serum creatinine (greater than 50 micromol/L) were compared with those that did not. RESULTS: Of the cases studied, 16/35 died perinatally, all showing anamnios and markedly elevated urinary electrolytes. Of the survivors, ten maintained normal postnatal serum creatinine, whereas nine did not. The frequency of reduced amniotic fluid/olygohydramnios was higher in cases that developed increased serum creatinine (four out of nine) than in those that did not (nil). Sodium above the 95th percentile was 100% specific and 44% sensitive to predict an increased serum creatinine during early infancy, while chloride above the 95th percentile was 70% specific and 56% sensitive. All seven cases in which urinary sodium was elevated and/or amniotic fluid volume was reduced developed renal failure. CONCLUSION: Urine sampling slightly improved renal function prediction, but this must be balanced against its fetal risks. PMID- 16374902 TI - Non-normal path analysis in the presence of measurement error and missing data: a Bayesian analysis of nursing homes' structure and outcomes. AB - Path analytic models are useful tools in quantitative nursing research. They allow researchers to hypothesize causal inferential paths and test the significance of these paths both directly and indirectly through a mediating variable. A standard statistical method in the path analysis literature is to treat the variables as having a normal distribution and to estimate paths using several least squares regression equations. The parameters corresponding to the direct paths have point and interval estimates based on normal distribution theory. Indirect paths are a product of the direct path from the independent variable to the mediating variable and the direct path of the mediating variable to the dependent variable. However, in the case of non-normal distributions, the point and interval estimates of the indirect path become much more difficult to estimate. We address the issue of calculating indirect point and interval estimates in the case of non-normally distributed data. Our substantive application is a nursing home research problem in which the variables in the path analysis of interest involve variables with normal, Bernoulli, or Poisson distributions. Additionally, one of the Poisson variables is observed with error. This paper addresses estimating point and interval estimation of indirect paths for variables with non-normal distributions in the presence of missing data and measurement error. We handle these difficulties from a fully Bayesian point of view. We present our substantive path analysis motivated from a nursing home structure, process, and outcomes model. Our results focus on the impact job turnover in the nursing homes has on nursing home outcomes. PMID- 16374903 TI - The power of the standard test for the presence of heterogeneity in meta analysis. AB - It has been suggested that the standard test for the presence of heterogeneity in meta-analysis has low power. Although this has been investigated using simulation, there is little direct analytical evidence of the validity of this claim. Using an established approximate distribution for the test statistic, a procedure for obtaining the power of the test is described. From this, a simple formula for the power is obtained. Although this applies to a special case, the formula gives an indication of the power of the test more generally. In particular, for a given significance level, the power can be calibrated in terms of the proportion of the studies' variances that is provided by between-study variation. A consideration of this quantity confirms that the test does, in general, have low power. It is suggested that practitioners, who wish to conduct the standard test, use the ideas provided in order to investigate the operating characteristics of the test prior to performing it. PMID- 16374904 TI - A sequential design to estimate sensitivity and specificity of a diagnostic or screening test. AB - We consider efficient study designs to estimate sensitivity and specificity of a candidate diagnostic or screening test. Our focus is the setting in which the candidate test is inexpensive to administer compared to evaluation of disease status, and the test results, available in a large cohort, can be used as a basis for sampling subjects for verification of disease status. We examine designs in which disease status is verified in a sample chosen so as to optimize estimation of either sensitivity or specificity. We then propose a sequential design in which the first step of sampling is conducted to efficiently estimate specificity. If the candidate test is determined to be of sufficient specificity, then step two of sampling is conducted to estimate sensitivity. We propose estimators based on this sequential sampling scheme, and show that the performance of these estimators is excellent. We develop sample size calculations for the sequential design, and show that this design, in most situations, compares favourably in terms of expected sample size to a fixed size design. PMID- 16374905 TI - Linear independence of pairwise comparisons of DNA microarray data. AB - MOTIVATION: For DNA microarrays, the gain in certainty by performing multiple experimental repeats is offset by the high cost of each experiment. In a typical experiment, two independent measurements (that is, data from two separate arrays) are combined to yield a single comparative index per gene. Thus, although one uses 2n arrays and performs 2n independent measurements, one obtains only n comparative measurements. We addressed the question: how many of the potential n2 comparisons derivable from such data are actually independent, and what effect do these additional comparisons have on the false positive rate. RESULTS: We show there are precisely 2n - 1 independent comparisons available from among the n2 possibilities. Applying these additional n - 1 independent comparisons to experimental and simulated data reduced the false positive rate by as much as 10 fold, with excellent agreement between experimental and theoretical false positive rates. PMID- 16374906 TI - Discovery of highly differentiative gene groups from microarray gene expression data using the gene club approach. AB - MOTIVATION: It is commonly believed that suitable analysis of microarray gene expression profile data can lead to better understanding of diseases, and better ways to diagnose and treat diseases. To achieve those goals, it is of interest to discover the gene interaction networks, and perhaps even pathways, underlying given diseases from such data. In this paper, we consider methods for efficiently discovering highly differentiative gene groups (HDGG), which may provide insights on gene interaction networks. HDGGs are groups of genes which completely or nearly completely characterize the diseased or normal tissues. Discovering HDGGs is challenging, due to the high dimensionality of the data. RESULTS: Our methods are based on the novel concept of gene clubs. A gene club consists of a set of genes having high potential to be interactive with each other. The methods can (i) efficiently discover signature HDGGs which completely characterize the diseased and the normal tissues respectively, (ii) find strongest or near strongest HDGGs containing any given gene, and (iii) find much stronger HDGGs than previous methods. As part of the experimental evaluation, the methods are applied to colon, prostate, ovarian, and breast cancer, and leukemia and so on. Some of the genes in the extracted signature HDGGs have known biological functions, and some have attracted little attention in biology and medicine. We hope that appropriate study on them can lead to medical breakthroughs. Some HDGGs for colon and prostate cancers are listed here. The website listed below contains HDGGs for the other cancers. AVAILABILITY: HDGG is implemented in C++ and runs on Unix or Windows platform. The code is available at: http://www.cs.wright.edu/~gdong/hdgg/. PMID- 16374907 TI - In silico phenotypic screening method of mutants based on statistical modeling of genetically mixed samples. AB - In comprehensive functional genomics projects, systematic analysis of phenotypes is vital. However, conventional phenotypic screening is done mainly by imprecise visual observation of qualitative traits, and, therefore, in silico screening techniques for quantitative traits are required. In this report, we propose in silico phenotypic screening method that utilizes a Gaussian mixture model for the trait distribution in the offspring of a mutagenized line and the likelihood ratio test between the estimated Gaussian mixture model and the wild-type single Gaussian model. In order to evaluate the proposed method, we performed a screening experiment using real trait data of Arabidopsis. In this experiment, the proposed screening method properly distinguished the mutant line from the wild-type line. Furthermore, we conducted power analysis of the proposed method and two conventional methods under various simulated conditions of sample size and distribution of trait frequency. The result of the power analysis confirmed the effectiveness of the proposed method compared to the conventional methods. PMID- 16374908 TI - Utilizing evolutionary information and gene expression data for estimating gene networks with bayesian network models. AB - Since microarray gene expression data do not contain sufficient information for estimating accurate gene networks, other biological information has been considered to improve the estimated networks. Recent studies have revealed that highly conserved proteins that exhibit similar expression patterns in different organisms, have almost the same function in each organism. Such conserved proteins are also known to play similar roles in terms of the regulation of genes. Therefore, this evolutionary information can be used to refine regulatory relationships among genes, which are estimated from gene expression data. We propose a statistical method for estimating gene networks from gene expression data by utilizing evolutionarily conserved relationships between genes. Our method simultaneously estimates two gene networks of two distinct organisms, with a Bayesian network model utilizing the evolutionary information so that gene expression data of one organism helps to estimate the gene network of the other. We show the effectiveness of the method through the analysis on Saccharomyces cerevisiae and Homo sapiens cell cycle gene expression data. Our method was successful in estimating gene networks that capture many known relationships as well as several unknown relationships which are likely to be novel. Supplementary information is available at http://bonsai.ims.u-tokyo.ac.jp/~tamada/bayesnet/. PMID- 16374909 TI - Refinement of NMR-determined protein structures with database derived distance constraints. AB - The protein structures determined by NMR (Nuclear Magnetic Resonance Spectroscopy) are not as detailed and accurate as those by X-ray crystallography and are often underdetermined due to the inadequate distance data available from NMR experiments. The uses of NMR-determined structures in such important applications as homology modeling and rational drug design have thus been severely limited. Here we show that with the increasing numbers of high quality protein structures being determined, a computational approach to enhancing the accuracy of the NMR-determined structures becomes possible by deriving additional distance constraints from the distributions of the distances in databases of known protein structures. We show through a survey on 462 NMR structures that, in fact, many inter-atomic distances in these structures deviate considerably from their database distributions and based on the refinement results on 10 selected NMR structures that these structures can actually be improved significantly when a selected set of distances are constrained within their high probability ranges in their database distributions. PMID- 16374910 TI - Heuristic search in constrained bipartite matching with applications to protein NMR backbone resonance assignment. AB - The constrained bipartite matching (CBM) problem is a variant of the classical bipartite matching problem that has been well studied in the Combinatorial Optimization community. The input to CBM is an edge-weighted complete bipartite graph in which there are a same number of vertices on both sides and vertices on one side are sequentially ordered while vertices on the other side are partitioned and connected into disjoint directed paths. In a feasible matching, a path must be mapped to consecutive vertices on the other side. The optimization goal is to find a maximum or a minimum weight perfect matching. Such an optimization problem has its applications to scheduling and protein Nuclear Magnetic Resonance peak assignment. It has been shown to be NP-hard and MAX SNP hard if the perfectness requirement is dropped. In this paper, more results on the inapproximability are presented and IDA*, a memory efficient variant of the well known A* search algorithm, is utilized to solve the problem. Accordingly, search heuristics and a set of heuristic evaluation functions are developed to assist the search, whose effectiveness is demonstrated by a simulation study using real protein NMR backbone resonance assignment instances. PMID- 16374911 TI - A wavelet approach for the analysis of folding trajectory of protein Trp-cage. AB - Understanding how protein folds into a functional native structure is arguably one of the most challenging problems remaining in computational biology. Currently, the protein folding mechanism is often characterized by calculating the free energy landscape in terms of various reaction coordinates such as the fraction of native contacts, the radius of gyration, the RMS deviation from the native and so on. In this paper, we present a wavelet approach towards understanding the global state changes during protein folding. The approach is based on the wavelet analysis on the trajectories of various reaction coordinates to identify the significant intermediate states or structural motifs in the folding process. We demonstrate through an example protein Trp-cage that this approach extracts crucial information about protein folding intermediate states as well as the time correlation among these states. Furthermore, the current approach reveals a meaningful structural pattern that had been overlooked in previous works, which provides a better understanding of the folding mechanism as well as the limitation of the current force fields. PMID- 16374912 TI - Gene function prediction by a combined analysis of gene expression data and protein-protein interaction data. AB - Prediction of biological functions of genes is an important issue in basic biology research and has applications in drug discoveries and gene therapies. Previous studies have shown either gene expression data or protein-protein interaction data alone can be used for predicting gene functions. In particular, clustering gene expression profiles has been widely used for gene function prediction. In this paper, we first propose a new method for gene function prediction using protein-protein interaction data, which will facilitate combining prediction results based on clustering gene expression profiles. We then propose a new method to combine the prediction results based on either source of data by weighting on the evidence provided by each. Using protein protein interaction data downloaded from the GRID database, published gene expression profiles from 300 microarray experiments for the yeast S. cerevisiae, we show that this new combined analysis provides improved predictive performance over that of using either data source alone in a cross-validated analysis of the MIPS gene annotations. Finally, we propose a logistic regression method that is flexible enough to combine information from any number of data sources while maintaining computational feasibility. PMID- 16374913 TI - Covariation analysis of local amino acid sequences in recurrent protein local structures. AB - Local structural information is supposed to be frequently encoded in local amino acid sequences. Previous research only indicated that some local structure positions have specific residue preferences in some particular local structures. However, correlated pairwise replacements for interacting residues in recurrent local structural motifs from unrelated proteins have not been studied systematically. We introduced a new method fusing statistical covariation analysis and local structure-based alignment. Systematic analysis of structure based multiple alignments of recurrent local structures from unrelated proteins in representative subset of Protein Databank indicates that covarying residue pairs with statistical significance exist in local structural motifs, in particular beta-turns and helix caps. These residue pairs are mostly linked through polar functional groups with direct or indirect hydrogen bonding. Hydrophobic interaction is also a major factor in constraining pairwise amino acid residue replacement in recurrent local structures. We also found correlated residue pairs that are not clearly linked with through-space interactions. The physical constrains underlying these covariations are less clear. Overall, covarying residue pairs with statistical significance exist in local structures from unrelated proteins. The existence of sequence covariations in local structural motifs from unrelated proteins indicates that many relics of local relations are still retained in the tertiary structures after protein folding. It supports the notion that some local structural information is encoded in local sequences and the local structural codes could play important roles in determining native state protein folding topology. PMID- 16374914 TI - A robust algorithm for ratio estimation in two-color microarray experiments. AB - The reliability of the algorithms for ratio estimation in two-color microarray image analysis is very important, as these ratios build up the primary source of information for the subsequent analytical procedures (normalization, clustering, classification, etc). Although various algorithms already exist, there is still a need to develop procedures having higher levels of accuracy and robustness. We present a statistical procedure for the detection and removal of aberrant pixels in two-color microarray images. It is based on a linear regression approach, assuming reasonably high level of correlation between the two color channels. This procedure ensures more robust ratio estimation for the spots in both linear regression and traditional segmentation algorithms. The developed algorithms have been evaluated using simulated artificial images and experimental images of different designs. A demonstration version of the software can be downloaded from http://bioinfo.curie.fr/projects/maia/. PMID- 16374915 TI - A dimensionless fit measure for phylogenetic distance trees. AB - We present a dimensionless fit index for phylogenetic trees that have been constructed from distance matrices. It is designed to measure the quality of the fit of the data to a tree in absolute terms, independent of linear transformations on the distance matrix. The index can be used as an absolute measure to evaluate how well a set of data fits to a tree, or as a relative measure to compare different methods that are expected to produce the same tree. The usefulness of the index is demonstrated in three examples. PMID- 16374916 TI - A tutorial on Markov models based on Mendel's classical experiments. AB - Hidden Markov Models (HMM) can be extremely useful tools for the analysis of data from biological sequences, and provide a probabilistic model of protein families. Most reviews and general introductions follow the excellent tutorial by Rabiner, where the focus is outside biology. Mendel's famous experiments in plant hybridisation were published in 1866 and are often considered the icebreaking work of modern genetics. He had no prior knowledge of the dual nature of genes, but through a series of experiments he was able to anticipate the hidden concept and name it "Elemente". In this paper we present the background, theory and algorithms of HMM based on examples from Mendel's experiments, and introduce the toolbox "mendelHMM". This approach is considered to have some intuitive advantages in a biological and bioinformatical setting. Applications to analysing bio-sequences like nucleic acids and proteins are also discussed. PMID- 16374917 TI - [Abstracts of the 21st Annual Congress of the European Society of Human Reproduction and Embryology, Copenhagen, Denmark, 19-22 June 2005]. PMID- 16374918 TI - Whose Bion? Who is Bion? PMID- 16374919 TI - Developmental delay. PMID- 16374920 TI - Bion: theoretical and clinical observations. PMID- 16374922 TI - Abstracts of the 28th Annual San Antonio Breast Cancer Symposium, December 8-11, 2005, San Antonio, Texas, USA. PMID- 16374921 TI - Unannounced standardised patients. PMID- 16374923 TI - Mandatory notification of child abuse. PMID- 16374924 TI - Duty of confidentiality. PMID- 16374926 TI - Survey of knowledge of popular children's TV characters among paediatric healthcare professionals. PMID- 16374925 TI - Trial registration, the ICMJE statement, and paediatric journals. PMID- 16374927 TI - First aid teaching for undergraduate medical students. PMID- 16374928 TI - Group study as a tool to teach medical students about coronary heart disease (CHD). PMID- 16374929 TI - Professionalism and medical education. PMID- 16374930 TI - Distribution of polychlorinated biphenyls in the water, sediment and fish from the Pearl River estuary, China. AB - The concentrations and distribution of polychlorinated biphenyls were determined in water, surface sediments and fishes from the estuaries of Pearl River, PR China. The results showed that the concentrations ranged from 2.47 ng l-1 to 6.75 ng l-1 in water, from 11.13 ng g-1 to 23.23 ng g-1 in sediments (dry weight) and from 68.64 ng g-1 to 316.85 ng-1 in fish (lipid weight). It was found that the feeding habits of fish were very significant in the accumulation of polychlorinated biphenyls and pattern of congeners. Carnivorous and benthic fishes such as eel (Anguilla japonica) and Chinese sea catfish (Arius sinensis) were found to have high concentrations of polychlorinated biphenyls and of high chlorinated congeners, while herbivores such as shad (Clupanodon punctatus) and mullet (Mugil cephalus) exhibited an opposite trend. Congeners IUPAC Nos. 153, 138, 118, 87/81, 170 and 52 were found frequently in most samples (both sediment and fish). In comparison with other places in the world, the concentration of polychlorinated biphenyls in sediment and fish from the estuary of Pearl River were low. PMID- 16374931 TI - Oklahoma Nurses Association 2005 House of Delegates resolution. Implications of fatigue on patient safety. AB - The ANA 2005 House of Delegates resolved to disseminate recent research findings of the relationship between work hours, fatigue and errors. During the House, Dr. Ann Rogers presented the, results of her study "The Working Hours of Hospital Staff Nurses and Patient Safety" published in Affairs, July/August 2004. Her study followed 393 registered nurses working over 5,317 shifts. Each nurse tracked hours worked, time of dayworked, overtime, days off, sleep/wake patterns, errors and near misses. Findings included correlation between work duration, overtime and number of hours worked, on reported errors and near errors. Regardless of the scheduled shift length, overtime increased the odds of reporting error, but the risks significantly increased when overtime followed a twelve-hour shift. Responsibilities for patient safety and fatigue rest with the individual nurse as well as employers. It is the responsibility of the nurse to obtain adequate sleep prior to coming to work, use caffeine judiciously and plan work toassure appropriate rest breaks. While it is the responsibility of the employing agency to provide for staffing, nurses should work collaboratively with managers and within their team to assure an appropriate schedule. Nurses have to make decisions about the total number of hours they work, and in the event of working voluntary overtime, it is the nurse's responsibility to assess the cumulative effects of long hours, and plan a schedule that assures our responsibility provide safe, effective care. PMID- 16374932 TI - Patient safety alert. Health care workers may imperil patients by snubbing influenza vaccinations. PMID- 16374933 TI - Nationwide solutions could come from Massachusetts' uninsured problem. PMID- 16374934 TI - CA: RN tries to amend suit to 'class action': court refuses to allow class action status. PMID- 16374935 TI - Quality of life after ileal pouch-anal anastomosis and ileorectal anastomosis in patients with familial adenomatous polyposis. AB - PURPOSE: Differences in conventional outcomes such as functional results and the rate of complications have caused a controversy about whether the ileal pouch anal anastomosis or the ileorectal anastomosis is the preferred surgical treatment for patients with familial adenomatous polyposis. We therefore sought to ascertain not only the surgical results but also the perceptions of patients about their outcomes. METHODS: Between 1981 and 1998, 152 patients at our institution had an ileal pouch-anal anastomosis and 32 patients had an ileorectal anastomosis for familial adenomatous polyposis. Of these 184 patients, 173 were sent a study-specific quality-of-life questionnaire and the Short Form 36 health survey to determine their health-related quality of life. RESULTS: Ninety-four ileal pouch patients and 21 ileorectal patients returned the surveys. No difference was found in early postoperative complications, 5-year probability for complications, or functional results after either procedure. On the Short Form 36 health survey, the ileorectal patients had a lower mental health summary score compared with that of the ileal pouch patients but a similar physical health summary score. The study-specific questionnaire found both groups to have a comparable quality of life. CONCLUSION: Because ileal pouch-anal anastomosis has the advantage of removing as much at-risk tissue as possible with similar functional results and better mental health, it may be considered the preferred operation for most patients with familial adenomatous polyposis. PMID- 16374936 TI - Mechanisms of microsatellite instability in colorectal cancer patients in different age groups. AB - PURPOSE: The proportion of colorectal cancers located proximal to the splenic flexure increases with age. Colorectal cancers of the microsatellite instability phenotype are preferentially located in the proximal colon. We investigated the location of colorectal cancer with this phenotype in different age groups to determine whether different molecular mechanisms could account for the changes in distribution of colorectal cancers. METHODS: A representative sample of 230 colorectal cancers from three age groups (<45 years, 60-70 years, >87 years) was selected from a subset of The Upper Midwest Oncology Medical Registries database. Microsatellite instability was determined by polymerase chain reaction using a panel of five microsatellite markers. The presence of new microsatellite alleles at two or more loci was scored as microsatellite instability. Tumors were otherwise considered microsatellite stable. MLH1 and MSH2 expression was determined by immunohistochemistry. Methylation of the MLH1 gene promotor was determined by methylation-specific polymerase chain reaction assay. RESULTS: The proportion of tumors of the microsatellite instability phenotype was 21 percent in the young group, 15 percent in the middle group, and 33 percent in the old group. More tumors of the microsatellite instability phenotype were proximal compared with microsatellite-stable tumors in all three age groups, but the differences were significant only for the old group. Tumors of the microsatellite instability phenotype in the older group were associated with MLH1 inactivation (24/29 or 83 percent), MLH1 promoter methylation (18/29 or 62 percent), and proximal location (25/29 or 86 percent), while tumors in the young group were associated with MSH2 inactivation (8/18 or 44 percent) and distal location (11/18 or 62 percent). CONCLUSION: The age-related proximal shift of colorectal cancers is associated with the microsatellite instability phenotype, MLH1 inactivation, and MLH1 promoter hypermethylation. PMID- 16374937 TI - Fibrous glass insulation and cancer: response and rebuttal. AB - In response to our commentary on fibrous glass and cancer [Infante et al., 1994], three letters have been received by the journal. The arguments put forth in these letters do not lead us to alter our scientific view that fibrous glass insulation is carcinogenic. No information is given in the letters that has not previously been stated. Even though these letters raise the same diaphanous arguments, we believe that to preserve occupational and public health we must respond in detail to ensure that the contentions of the fibrous glass industry do not gain further acceptance. Thus, we respond to each letter in turn: The first by Weiss questions and distorts epidemiologic findings, the second by McConnell attempts to recant his past views on the carcinogenicity of fibrous glass and on the value of rodent bioassays in general, and the third by Hesterberg and Chase impugns our analyses demonstrating a positive cancer response in their study. Lastly, we have not debated every issue raised in these three letters because of space limitations, and have centered our responses on what we consider the major incongruities and falsities in each letter. Likewise, we have been selective in citing only relevant literature, or those reports used by the industry or its consultants to support their views. PMID- 16374938 TI - Inappropriate care--professional misconduct (02/96C). PMID- 16374939 TI - [Risk management--adverse effects of medication: auditory toxicity and adverse effects of steroid preparations]. PMID- 16374940 TI - Use of continine immunoassay test strips for preclassifying urine samples from smokers and nonsmokers prior to analysis by LC-MS-MS. AB - Cotinine biomarker measurements involving both smokers and nonsmokers must accommodate a broad range of concentrations. Thus, we have routinely preclassified unknown samples as being either "high" or "low" by using an enzyme linked immunoassay for cotinine prior to analysis by tandem mass spectrometry (MS). Although this method is effective, it is also time-consuming and complex; a simpler and faster approach would be useful. Consequently, a screening assay for urine cotinine using an immunochromatographic test strip (NicAlert) followed by a computerized analysis of the data was examined as a possible alternative. The results indicate that this approach can provide useful classification efficiency when using our target cutoff value of approximately 20 ng/mL. In the analysis of 50 urine samples from nonsmokers with varying degrees of exposure to environmental tobacco smoke, the classification sensitivity and specificity were 88% and 92%, respectively, for cotinine measured by the test strips relative to total cotinine concentrations measured by atmospheric-pressure ionization tandem MS. However, the relatively high cost of the strips may be a limiting factor. PMID- 16374941 TI - Automated solid-phase extraction and quantitative analysis of 14 phthalate metabolites in human serum using isotope dilution-high-performance liquid chromatography-tandem mass spectrometry. AB - Phthalates are industrial chemicals with many commercial applications. Because of their common usage, the general population is exposed to phthalates. A sensitive and selective analytical method is necessary to accurately determine the phthalate levels in serum. We improved our previously developed analytical method to measure nine phthalate metabolites in human serum by automating the solid phase extraction (SPE) procedure and by including five additional phthalate metabolites: phthalic acid; mono-isobutyl phthalate, a metabolite of di-isobutyl phthalate; mono-(3-carboxypropyl) phthalate, a major oxidative metabolite of di-n octyl phthalate; and mono-(2-ethyl-5-oxohexyl) phthalate and mono-(2-ethyl-5 hydroxyhexyl) phthalate, two oxidative metabolites of di-(2-ethylhexyl) phthalate. Automation of the SPE eliminated the human variation associated with the manual SPE, thus improving the reproducibility of the measurements. Additional wash steps during SPE produced cleaner extracts and resulted in higher recoveries (80-99%) than the manual SPE method. Furthermore, the automated SPE method allowed for the unattended extraction of samples, with a concomitant increase in sample throughput compared to the manual SPE method. The method is accurate, precise, and sensitive, with limits of detection in the low nanogram per-milliliter range. PMID- 16374942 TI - Can an immunoassay become a standard technique in detecting oxycodone and its metabolites? AB - Opiate toxicology testing is routinely performed in the hospital setting to identify abusers and/or to determine those patients who are not taking prescribed opiate analgesics such as oxycodone. Commercially available assays for opiate detection in urine have decreased sensitivity for oxycodone, which contributes to a high false-negative rate. Functioning as a beta site, our Veterans Affairs hospital evaluated a new enzyme immunoassay, DRI Oxycodone Assay, for its use in the qualitative and semiquantitative detection of oxycodone in urine. We hypothesize that an immunoassay for oxycodone with superior sensitivity and specificity, when compared to the traditional opiate assays, would reduce the need for more expensive and time-consuming confirmatory testing. We used the new liquid homogenous enzyme immunoassay to determine oxycodone results in a total of 148 urine samples from 4 different sample groups. Gas chromatography-mass spectroscopy was subsequently used to confirm the presence or absence of oxycodone (or its primary metabolite, noroxycodone). We also evaluated within run, between-run, and linearity studies and conducted a crossover study to establish a cutoff value for oxycodone. In our patient population, we used the new DRI immunoassay to evaluate 17,069 urine samples to estimate oxycodone misuse profiles (patients not taking prescribed oxycodone or taking oxycodone without a prescription) during a 4-month period. The sensitivity and specificity of the new oxycodone immunoassay were 97.7% and 100%, respectively, at the cutoff concentration of 300 ng/mL. The assay linearity was 1,250 ng/mL, and the sensitivity was 10 ng/mL. Within-run precision and between-run coefficient of variation were 2.3% and 1.8%, respectively. None of the 15 compounds that we evaluated for interference had crossover significant enough to produce a positive oxycodone result when using 300 ng/mL as the cutoff value. None of the 17,069 oxycodone immunoassays was followed with a request for confirmation. Among patients with positive results (n = 224), 93 (41.5%) were not prescribed oxycodone. The new DRI Oxycodone Assay is a sensitive and specific screening test for the determination of oxycodone. The improved opiate screening results may lead to better patient and prescription management, to decreased laboratory spending, and to the identification of oxycodone abusers, which could result in decreased oxycodone-related mortality. PMID- 16374943 TI - The complex analytical method for assessment of passive smokers' exposure to carbon monoxide. AB - Carbon monoxide (CO) is one of the toxic constituents in tobacco smoke. The aim of the study was to evaluate a complex analytical method that allows quantification of the exposure of passive smokers to CO. The exposed volunteers were placed in the model room where portions of cigarettes (5, 10, or 15 cigarettes every 30 or 60 min over 4 h) were smoked using a homemade smoking machine. The concentrations of CO and of the ventilation marker (methane) were monitored for the duration of each experiment. CO and methane were analyzed on line using GC-FID with methanization. Carboxyhemoglobin (COHb) was analyzed twice: just before and after the experiment. After hemolysis of a 100-microL blood sample, CO was quantitatively released by adding 10% K3[Fe(CN)6] inside a small reactor and under stable pressure transported through a microtube with an absorbing agent on a chromatography loop. The proposed analytical method allows us to quantify exposure of passive smokers by measuring the dose-effect correlation. We observed that increasing COHb levels were directly correlated with the CO concentration in the air, but were also intermediately correlated with the frequency and number of smoked cigarettes and with the ventilation coefficient. PMID- 16374944 TI - Detection of recombinant epoetin and darbepoetin alpha after subcutaneous administration in the horse. AB - A direct detection method for anti-doping control of recombinant human erythropoietin (rHuEPO) abuse in racehorses is proposed. This method involves screening of plasma (or serum) by an enzyme-linked immunosorbent assay specific for human EPO and confirmation in urine samples by characterization of the urinary EPO isoelectric profile. This method was tested on horses that were administered epoetin alpha (rHuEPO) and the hyper-glycosylated form of this drug (darbepoetin alpha). PMID- 16374945 TI - Differences in incidence and distribution of colorecetal cancer among races and ethnic societies: lifestyle, genes or both? PMID- 16374946 TI - Chronic kidney disease. PMID- 16374947 TI - Need equipment? Think eBay. PMID- 16374948 TI - Submitting Medicare claims. PMID- 16374950 TI - The ambush. PMID- 16374949 TI - Pay for performance. How fast is it spreading? PMID- 16374951 TI - Refusing to settle could cost you. PMID- 16374952 TI - The exam room is my sanctuary. PMID- 16374953 TI - Hands healing hearts and bodies. PMID- 16374954 TI - Is your lawyer charging too much? PMID- 16374955 TI - Selling our practice: a real roller-coaster ride. PMID- 16374956 TI - You can get home in time for dinner. PMID- 16374957 TI - Do disease registries = dollar rewards? PMID- 16374958 TI - A profile in courage. PMID- 16374959 TI - Adding ancillaries: boosting the bottom line. PMID- 16374960 TI - Laser hair removal. PMID- 16374961 TI - Getting insurers to pay up. PMID- 16374962 TI - Taking cross-training too far. PMID- 16374964 TI - A letter from the president. PMID- 16374963 TI - Reflections on hurricanes Katrina and Rita. PMID- 16374965 TI - Hurricane Katrina: one doctor's journal. PMID- 16374966 TI - ECG of the month. Hypotension, pulmonary edema, and an irregular cardiac rhythm in a 50-year-old woman. Sinus tachycardia, type I (Wenckebach) second degree atrioventricular block, acute inferoposterior myocardial infarct, and anterolateral myocardial infarct of indeterminate age (probably old). AB - A 50-year-old woman came to the emergency department because of chest discomfort and dyspnea. She was found to have hypotension, oliguria, and pulmonary edema, i.e., full-blown cardiogenic shock, an irregular rhythm, and no cardiac murmur. The electrocardiogram (ECG) was recorded one lead at a time, and a lead II rhythm strip was mounted above the standard 12 leads (Figure 1). PMID- 16374967 TI - Is this a case of simple soft tissue mass? AB - A 15-year-old girl reports a 1-year history of a mass on the lower portion of her left leg associated with occasional pain. Antero-posterior and lateral radiography, T1 and T2 magnetic resonance imaging (MRI), and bone/joint whole body scan were performed. PMID- 16374968 TI - Fever, chills, and weakness in a 61-year-old man. AB - A 61-year-old man presented to the emergency department of a community hospital with a 2-week history of fever, chills, and sudden extreme weakness of his right arm and lower extremities. He also had a cough, shortness of breath, nausea, abdominal pain, diarrhea, and myalgia. Though initially alert and cooperative, he quickly became unresponsive. In addition, he had hyponatremia, renal insufficiency, and compromised cardiopulmonary function. He was admitted to the intensive care unit for suspected bacterial infection and was started on broad spectrum antibiotics. Chest radiograph revealed miliary infiltrates consistent with infectious emboli or metastatic carcinoma. Despite intensive resuscitation, the patient died 36 hours after admission. At autopsy multiple nodular lesions were observed on gross examination of the lungs, perihilar and paratracheal lymph nodes, and liver. Microscopic sections of the lung (Figure 1) and brain (Figures 2 and 3) are shown. PMID- 16374969 TI - A young man with back and chest wall pain for three months. PMID- 16374970 TI - Intracapsular versus standard tonsillectomy: review of literature. AB - For the past century, standard (total) tonsillectomy has been the preferred method for treating illness resulting from tonsillar infection and hypertrophy. With the advent of antibiotics, however, more cases of tonsillitis are treated medically. As a result, tonsillectomies are being performed less frequently for infection than for obstructive symptoms associated with tonsillar hypertrophy and sleep-disordered breathing. This shift has led to the re-emergence of the tonsillotomy, and more specifically the partial or intracapsular tonsillectomy, as an option for treating tonsillar hypertrophy, mainly in an effort to reduce the post-operative complications of pain, subsequent dehydration from lack of oral intake, and, potentially, delayed postoperative bleeding. The following is a review of the literature comparing intracapsular tonsillectomy to standard tonsillectomy for tonsillar hypertrophy associated with sleep-disordered breathing. PMID- 16374971 TI - Prevalence of overweight and obesity within primary care clinics in a public hospital system. AB - OBJECTIVE: The prevalence of obesity is especially high in low-income minority individuals, many of whom lack health insurance. The goal of the current study was to establish the prevalence of obesity in the primary care clinics at a public hospital that serves predominantly African-American indigent patients and to compare this prevalence to the national norm. RESEARCH METHODS: Information from 845 patients attending four outpatient clinics was obtained via retrospective chart reviews. Age, gender, race, weight, and height were obtained, and body mass index (BMI) was calculated. RESULTS: Eighty-four percent of patients were overweight or obese. The 20% prevalence of extreme obesity was especially high. The primary care clinics had more than twice the rate of obesity as the national norm, even when controlling for demographic characteristics. DISCUSSION: Socioeconomic status and/or cultural influences are likely factors in the higher rate of obesity in this population. Because of this markedly elevated prevalence of obesity, especially extreme obesity, the need for intervention is critical. PMID- 16374972 TI - Surveillance for West Nile virus cases in Louisiana 2001-2004. AB - West Nile virus (WNV) was first detected in Louisiana during August of 2001. An outbreak of 204 human cases of neuro-invasive disease (NID) and 25 deaths occurred in 2002. In the 2 years following, lower numbers of human cases were identified (101 NID cases in 2003 and 84 in 2004) but intense localized foci were observed. The incidence of NID has been particularly high in the elderly (65 years and older). The distribution of West Nile cases has consisted of sporadic cases with a few very intense foci. Annually, human cases have occurred from June through December, with a peak number of new cases in August. As compared with other WNV serosurveys conducted in the United States, it appears that the WNV seroprevalence in Louisiana is not elevated. PMID- 16374973 TI - Life-threatening occult gastrointestinal hemorrhage from a late complication of laparoscopic cholecystectomy. PMID- 16374974 TI - Should the NHS revise its role in medical student education? PMID- 16374975 TI - Clinical governance: what does it mean for NHS Scotland? PMID- 16374976 TI - I'm sorry Mr Paxman! PMID- 16374977 TI - Acute headache and subarachnoid haemorrhage: a retrospective review of CT and lumbar puncture findings. AB - AIM: Assess and correlate the clinical presentation, cranial CT and lumbar puncture (LP) findings in patients presenting with acute headache, clinically suspicious of subarachnoid haemorrhage, from the Accident and Emergency (A/E) department of a main teaching hospital. METHOD: We retrospectively reviewed over a 1-year period all patients referred for cranial CT from the A/E department, with clinically suspected non-traumatic SAH. Patients with a negative CT, with a diagnosis of SAH on LP, had their initial CT examination reassessed in a double blind review. RESULTS: 116 consecutive patients were included in the study. Patients were divided into 3 groups post CT results: diagnostic of SAH (19), abnormal without evidence of SAH (16) and normal (81). The clinical management of the latter group was analysed with particular emphasis on the performance and results of lumbar puncture (LP). 81 patients, clinically suspicious of SAH in our study had a normal cranial CT and were eligible for LP 49% had an LP, which was positive in 15% and contributed 24% to the total number diagnosed with SAH. LP was not performed in 51% of eligible patients. CONCLUSION: This study reviews the varying clinical presentations of SAH and the difficulty in clinically diagnosing SAH where headache is the only symptom. We review the wide differential diagnosis on cranial CT and on discharge of patients presenting with symptoms clinically suspicious of SAH. In addition this study identfies a high proportion of patients without detailed clinical notes and discharged without a diagnosis. The study reaffirms that a significant proportion of patients are diagnosed on LP and stress the importance of performing a LP in all cases with clinically suspected SAH with a negative CT This latter point requires highlighting in the medical literature as over half of eligible patients did not have a LP performed despite this being accepted diagnostic protocol. PMID- 16374978 TI - Gender inequalities in the management of angina pectoris: cross-sectional survey in primary care. AB - BACKGROUND AND AIMS: To determine the extent of gender differences in the routine clinical care of patients with angina pectoris in primary care. METHODS: A cross sectional survey of general practitioner (GP) medical records undertaken by trained data managers in 6 GP practices. 925 adults (489 men) with a clinical diagnosis of angina (prevalence = 2.4%, 95%CI 2.3-2.6). Data extracted included: level of care; risk factor recording; prescribed medication; exercise ECG and coronary revascularisation. Adjusted male-to-female odds ratios (AOR) adjusted for age, angina duration, and previous myocardial infarction, (MI). RESULTS: Women with angina were older than men (71 v 65 years) with a lower prevalence of MI (30% v 45%), but a longer duration of angina (5 v 4 years). Men were more likely to receive once daily aspirin (AOR = 2.07, 95%CI 1.56-2.74) and be prescribed triple anti-anginal therapy (1.58, 95%CI 1.03-2.42). Men were also significantly more likely to undergo exercise ECG (1.56, 95%CI 1.14-2.15) and surgical revascularisation (1.71, 95%CI 1.03-2.85). Women tended to receive GP care alone (AOR =0.64, 95%CI 0.46-0.89), whilst men received specialist cardiac care (1.47, 95%CI 1.09-2.00). Beta-blocker use following MI was similar (0.99, 95%CI 0.59-1.69). CONCLUSION: Differences in the management of men and women are unaccountedfor by differences in age, previous MI or duration of angina. Gender differences in management of CHD reported from secondary care may also exist in primary care. PMID- 16374979 TI - Do creatine kinase (CK) levels influence the diagnosis or outcome in patients with compartment syndrome? AB - We report a retrospective study in patients with compartment syndrome with the aim to analyse: 1) whether the level of rise in CK levels is a useful marker to the presence of compartment syndrome and; 2) whether an early fasciotomy assists in lowering of the elevated CK levels. The results of our study suggest that although raised CK levels are not diagnostic, they are a useful adjunct in making a diagnosis, and hence CK estimation should be done in all patients with suspected compartment syndrome. Moreover, an early fasciotomy (< 12 hours) has a statistically significant influence in lowering elevated CK levels, confirming the view that the earlier the decompression, the lesser the muscle damage. PMID- 16374980 TI - Conservative management, percutaneous or open repair of acute Achilles tendon rupture: a retrospective study. AB - BACKGROUND AND AIMS: To ascertain the differences in patients' perceived outcomes between conservative, percutaneous or open repair of Achilles tendon ruptures. METHODS: We studied 111 patients who had been managed for a unilateral closed Achilles tendon rupture. We excluded patients with open Achilles tendon lesions, patients whose tear had occurred more than seven days from operation, patients with diabetes, inflammatory disease, systemic corticosteroids or fluoroquinolones. Patients were contacted by telephone and asked to answer a questionnaire. RESULTS: There was no difference in effects on working life, patient satisfaction, sports performance, muscle strength, swelling and fear of re-rupture. Pain on weight bearing and cramps were signficantly more frequent in the patients managed conservatively. Re-rupture and complication rates were comparable to published rates. CONCLUSION: Treatment should be individualised according to the demands and health of the patient. PMID- 16374981 TI - Are atrial fibrillation guidelines altering management? A community based study. AB - BACKGROUND AND AIMS: We wanted to determine the prevalence of atrial fibrillation (AF) in a community based cross sectional study in greater Glasgow and how current anti-thrombotic management compares to published guidelines. METHODS: 1466 patients with AF were identified in General Practices in our community and 1008 consented to take part. Their demographic details and medical history were recorded. RESULTS: 1466 patients (mean age 73.4; 55% female) with AF were identified, in our community, giving a prevalence of 1%. 53% of patients were on warfarin therapy. Of those not receiving warfarin, only one third had a putative contra-indication. The proportion ofAF patients on warfarin increased with increasing stroke risk, and over the period of the study. CONCLUSIONS: Prevalence of AF was in keeping with previous estimates. The proportion of patients with AF receiving warfarin therapy appears to be increasing. In the moderate risk group, there was a tendency to use more warfarin in the younger age groups compared to the elderly. It was in the moderate and low risk groups that there was still evidence of deviation from published guidelines. PMID- 16374982 TI - Prescribing a website. AB - AIM: To assess the value of directing the attention of patients to sources of medical information on the internet. DESIGN: Prospective qualitative study in an orthopaedic outpatient clinic. PARTICIPANTS: 253 patients agreed to complete electronic questionnaires before and after reviewing information relevant to their conditions on the internet. Patients were allocated randomly into two groups; one group was given indications of general sites and the other recommended specific non-commercial sites. Completed questionnaires were received from 44 patients. RESULTS: 95% of the patients found the internet information easy to understand and 84% said that it was helpful for coping. 86% of the patients were satisfied that their current treatment was appropriate in the light of what they had learned from the internet. Ten patients out of the 36 who expressed a view thought that the internet information contradicted that provided by the doctor. Despite these results most patients still said that the doctor represented the best source of patient education. CONCLUSIONS: Increasing numbers of patients are familiar with the internet. Most of our patients felt that the internet was, on balance, helpful in providing information. The main difficulties with the internet are the sheer volume of information, the potential for misleading and the danger of misunderstanding. We feel that there is a real place for the specific prescription of an internet site by a clinician who has personally reviewed it to a patient thought to be able to benefit from it. PMID- 16374983 TI - Severe osteomalacia in a patient with idiopathic chronic pancreatitis. AB - We report a 30-year-old woman who was confined to a wheelchair because of severe myopathy. She was first seen by a neurologist because of a convulsive syndrome of unknown etiology when she was nine. She was started on anticonvulsive drugs but the drug was stopped when her serum calcium level was found to be very low. She had a history from childhood of steatorrhea and abdominal pain after a fatty meal and became vegetarian at age five years. She worked in a hospital as a nurse and at home her living room received no direct sunlight. As a result of these conditions osteomalacia progressed. We believe an awareness of chronic pancreatitis (CP) during childhood could have prevented the consequences of the disease in this case. PMID- 16374984 TI - Ossification of the Achilles tendon--a case report. AB - Ossification of the Achilles tendon is a very unusual condition. We report a case with bilateral Achilles tendon ossification followed up for twelve years, treated twice surgically on the left side and conservatively on the right side. At the age of 51, the patient's plain radiogram showed bilateral ossifications of the Achilles tendon (right 55 mm, left 15 mm in length). The left side small mass was removed. The right side mass was decided to be followed up because of its large size. At the age of 61, ossification recurred on the left side. Conservative treatment with etidronate disodium for two years failed to prevent ossification from progressing. At the age of 63, the mass on the right and left sides increased to 70 mm and 45 mm in length, respectively. The mass of the left side was excised again, and dull pain disappeared postoperatively on the left side. PMID- 16374985 TI - Cervical epidural abscess: an unusual case of persistent neck pain. PMID- 16374986 TI - Clinical skills teaching revisited. PMID- 16374987 TI - Potential costs of cardiac re-synchronisation therapy for chronic heart failure in Scotland. PMID- 16374988 TI - Cleaning the "hobby" closet. PMID- 16374989 TI - HAM it up. PMID- 16374990 TI - HAMs get involved. PMID- 16374991 TI - State of the art contemporary biomarkers of alcohol consumption. PMID- 16374992 TI - Transfusion safety: what has been done; what is still needed? PMID- 16374993 TI - Hospital finance: a primer for the 'wannabe' lab manager. PMID- 16374994 TI - Scholarship moves MLT toward dream to help Sudan. PMID- 16374995 TI - Natural disaster prompts rethinking responsibility. PMID- 16374996 TI - Blood transfusion: advocacy, adequacy, and safety. PMID- 16374997 TI - Born in the USA. PMID- 16374998 TI - Success or failure. PMID- 16374999 TI - Assessment tool. PMID- 16375000 TI - Consultant nurse networking. PMID- 16375001 TI - Crime and punishment. PMID- 16375002 TI - Prescribing partnership. PMID- 16375003 TI - Madelung deformity. PMID- 16375004 TI - Terrorism: the reality of blast injuries. PMID- 16375005 TI - A nurse led fast track service for patients with renal colic. PMID- 16375006 TI - Culture and care in the Swedish ambulance services. PMID- 16375007 TI - Board's eye view. PMID- 16375008 TI - Impacting legislation ranks as most important activity for Colorado Nurses Association. PMID- 16375009 TI - Steamboat Springs nurses respond to hurricane Katrina. PMID- 16375011 TI - Be a magnet, or just look like one. PMID- 16375012 TI - [Children with blunt trauma to the liver]. AB - In 3 children, 2 boys aged 13 years and an 8-year-old girl, blunt trauma to the liver was diagnosed after a fall. The first patient underwent drainage of abdominal fluid collection on multiple occasions and a stent was introduced in the biliary duct to ensure normal bile flow; the second patient was treated by arterial catheter embolisation of a segmental branch of the right hepatic artery; in the third patient, acute laparotomy was performed in view ofhaemodynamic instability and a large hepatic rupture was treated by packing. All patients recovered successfully. In the paediatric population, blunt trauma to the liver requires a multidisciplinary approach. For haemodynamically stable children, a conservative course can be taken. Arterial embolisation is the treatment of choice if there is a persistent need for volume resuscitation or blood transfusion. In case of massive haemorrhagic shock, the child should undergo laparotomy, and if the bleeding cannot be stopped, the liver should be packed according to damage-control principles. PMID- 16375013 TI - [Assessment, treatment and counselling of patients claiming sick leave and disability: recommendation from the Health Council]. AB - As requested by the Minister for Social Services and Employment, the Health Council of The Netherlands reviewed existing medical guidelines on the assessment, treatment and counselling of patients claiming sick leave and disability. These guidelines were developed separately by physicians concerned with their own specific fields. The Health Council recommends the development of more encompassing guidelines, based on scientific evidence whenever possible, starting with 10 diagnoses that frequently lead to long-term industrial disability. The assessment of long-term disability is complex and varies for each individual. The contribution of scientific 'evidence' remains modest. It is not possible to predict permanent disability with lists of diseases and corresponding medical prognoses. It may be possible to support the assessment by 'mediprudence', cases of example. The recommendation of the Health Council offers perspective toward qualitatively improved assessment of the effects of diseases on patient functioning. PMID- 16375014 TI - [Performance indicators as a measure of the quality of medical care: rhetoric and reality]. AB - Performance indicators may provide an indication of insufficient quality of medical care but they do not identify the cause of the problem. The political context for performance indicators is based on market ideology, where quality improvement is the goal and consumer pressure is the means. It is usually difficult to compare performance indicators among hospitals given the differences in definitions, methods of assessment, case mix, preclinical factors and data quality. Performance indicators are risk assessments and therefore subject to chance variation. Conclusions regarding performance indicators cannot be drawn due to the lack of clear, predefined benchmarks. At this time, the presence and degree of consumer pressure is unclear, and the ultimate effects of making performance factors publicly available on the quality of care is unknown. The question is whether mandatory reporting of a set of performance indicators by hospitals can provide sufficient insight into the quality of care; the measure appears to be too rough and too many factors influence the outcome. Procedure assessment is a good alternative to the use of performance indicators. PMID- 16375015 TI - [Clinical reasoning and decision-making in practice. A patient with oliguria following prostatectomy]. AB - A 70-year-old man with clinically localised prostate carcinoma underwent extraperitoneal endoscopic radical prostatectomy. His medical history revealed hypertension, renal colic, hypogonadotropic hypogonadism and recurrent deep venous thrombosis in the legs. The operation was uneventful with 500 ml blood loss and no periods ofhypotension. The patient developed oliguria within 12 h after surgery. A hypovolemic state was initially suggested to explain the oliguria and increasing amounts of intravenous fluids were administered. The oliguria persisted, however, and the patient did not respond to a diuretic. There was no fluid loss in the drain. Blood pressure, pulse and temperature were normal. Peritonitis and bowel perforation were excluded. Ultrasound examination of the bladder and kidneys revealed an empty bladder and no dilatation of the upper urinary tract, which excluded a post-renal obstruction. The clinical situation deteriorated within hours as the patient developed anuria, bowel distension, metabolic acidosis with progressive renal failure and signs of respiratory distress for which mechanical ventilation was needed. A chest X-ray prior to intubation did not show pneumonia or signs indicating pulmonary embolism. CT of the abdomen was performed to evaluate urinary leakage but revealed no fluid collection or urinoma. Thus pre- and post-renal causes of oliguria were excluded. In view of the systemic symptoms, intra-abdominal pressure was measured using a bladder catheter; it varied between 25 and 35 cm water. Together with the clinical situation, a diagnosis of abdominal compartment syndrome was made and coeliotomy was performed immediately. Within 10 min after decompression of the peritoneal cavity, diuresis started spontaneously. Renal function was restored to preoperative levels in 3 weeks. Abdominal compartment syndrome is a potentially life-threatening cause of anuria. The syndrome should be part of the differential diagnosis for patients with postoperative anuria, including those who underwent extraperitoneal minimally invasive procedures. PMID- 16375017 TI - [From gene to disease; neonatal diabetes mellitus and the KCNJ11 gene]. AB - Neonatal diabetes mellitus (DM) is by definition diagnosed within the first 3 months of life and can be either transient (TNDM) or permanent (PNDM). Recently, activating mutations in the KCNJ11 gene, which encodes the Kir6.2 subunit of the K(ATP) channel, have been identified as a cause of TNDM, the main cause of PNDM, and the cause of a new syndrome: developmental delay, epilepsy and neonatal diabetes. Patients with neonatal DM are normally dependent on life-long insulin injections, but patients with neonatal DM due to a KCNJ11 mutation are able to achieve control with sulphonylurea tablets. The mutations are predominantly spontaneous but have also been described as due to autosomal dominant inheritance and paternal mosaicism. Mutations at codon 201 and 59 are thus far the most prevalent. Because mutated K(ATP) channels do not close in response to ATP, the beta-cell membrane is hyperpolarised and insulin secretion does not occur. Mutated K(ATP) channels in muscle, nerve and brain are responsible for the neurological symptoms. PMID- 16375016 TI - [Revised guideline 'Vasectomy' from the Dutch Urological Association]. AB - Vasectomy is a simple and reliable method of contraception. Problems associated with vasectomy include inadequate patient information, complications of the procedure e.g. infection and scrotal bleeding (4-22%), chronic scrotal pain after the procedure (2-5%) and spontaneous recanalisation with return of fertility (0.03-I12%). Later in life a substantial number of men come to regret having a vasectomy, notably those who underwent it at a young age and those without children of their own. After 10 years 2.4% of vasectomised Dutch men have a refertilisation procedure (usually a vasovasostomy) because of the wish for children in a new relationship. Since vasectomy is an elective procedure and not done on medical indication, it requires an extensive informed-consent procedure for the patient. Insufficient information may result in inadequate follow-up, omission of semen analysis, and consequent legal procedures should complications or pregnancy ensue. Clearance after the first semen analysis at 3 months can be given if azoospermia is seen or if less than 100,000 non-motile spermatozoa are present in the ejaculate. PMID- 16375018 TI - [Diagnostic image (253). A man with a swollen penile base and scrotum]. AB - A 29-year-old man heard a snap during sexual intercourse followed by immediate detumescence and a swelling of the penile basis and scrotum, due to a penile fracture. PMID- 16375019 TI - [Increase in the number of invasive Haemophilus influenzae type b infections]. AB - OBJECTIVE: To describe the increase of invasive Haemophilus influenzae type b (Hib) infections in The Netherlands before and after the introduction of Hib vaccination in 1993, and to hypothesise about possible explanations. DESIGN: Descriptive. METHOD: Data on the prevalence of invasive Hib infections, such as meningitis and epiglottitis, during 1990-2004 were obtained from The Netherlands Reference Laboratory for Bacterial Meningitis, which collects Hib isolates from spinal fluid and blood from across the country. RESULTS: The incidence of invasive Hib infections decreased substantially for a few years after 1993. The total number of isolates was at a minimum in 1999 (n = 12) and increased to 49 in 2004. The annual number of patients with vaccine failure was 5 or less during 1995-2001, but was between 10 and 15 from 2002 onwards. A definite explanation for the increase in the incidence of invasive Hib infections cannot be given. Improbable causes are a surveillance artefact, an impaired response to the vaccine due to vaccination-scheme changes or interaction with other vaccines, or selection of Hib variants that are less sensitive to the vaccine-induced immunity. It most likely involves secondary vaccine failure: Hib carriership is decreased by mass vaccination, whereupon natural boosting occurs less frequently later in life. Subsequently, immunity decreases and susceptibility to invasive infection increases. Careful surveillance of invasive Hib infections in The Netherlands remains important. PMID- 16375020 TI - [Microsurgical vasovasostomy at the Erasmus MC, 1998-2002: results and predictive factors]. AB - OBJECTIVE: To determine the results ofmicrosurgical vasovasostomy procedures with special emphasis on the technical aspects and predictive factors for successful procedures. DESIGN: Descriptive. METHOD: In 1998-2002, 217 vasovasostomy procedures were performed in an outpatients' clinic setting at the Erasmus MC, Rotterdam, The Netherlands. RESULTS: Refertilisation was successful in 77% of men and 42% of couples reported a spontaneous pregnancy within one year. The most important predictive factors for a successful outcome were the interval between vasectomy and reversal and the age of the partner. Patency rates were 89% and 62% when the interval was < 5 years and > 15 years, respectively. Rates of spontaneous pregnancy were 56% and 21%, respectively. These reductions in rates of patency and pregnancy were attributed primarily to epididymal dysfunction and an increasing interval between vasectomy and reversal. Only 20% of men with a partner aged > 35 years reported spontaneous pregnancies. The risk of secondary epididymal obstruction was 25% after an interval of > 10 years. For these cases, a vasoepididymostomy is needed to achieve patency. PMID- 16375022 TI - [Assessment of medical competence in clinical education]. AB - There has been considerable change in the field of assessment of medical competence. At the moment, competency-orientated assessment, 'mini-CEX' (brief clinical evaluation exercises) and portfolios are quite popular. These methods are based on research findings indicating that medical competence can better be described as a collection of the complex tasks (so-called competencies) that a doctor must be able to perform than as the sum of knowledge, skills, problem solving ability and attitudes. Mini-CEX represents a method for the assessment of medical competence reliably and validly in a practical setting. Using a portfolio, information on the student's competence can be collated and evaluated from various sources, including mini-CEX. As such, a portfolio has much in common with a patient chart. PMID- 16375021 TI - [Pituitary apoplexy: an endocrinologic emergency]. AB - A 32-year-old woman, a 73-year-old man and a 26-year-old pregnant woman presented with headache, vomiting, and variable presence of visual disturbances, impaired consciousness, and circulatory shock. All three had pituitary apoplexy. In the first patient lymphocytic hypophysitis was diagnosed, the second had a nonfunctional adenoma and the last patient probably also. All three patients were treated conservatively with full recovery of signs and symptoms. However pituitary insufficiency remained in all three. Pituitary apoplexy is an acute event with significant morbidity and mortality. The cause is a rapid expansion of, usually, a pre-existing, often not yet known, adenoma by massive haemorrhage or infarction. Pressure, among other things, causes hypopituitarism, meningism, compression of the chiasma opticum and visual disturbances. Although many predisposing factors have been identified, pituitary apoplexy is often an unpredictable event. Diagnosis is made by the typical clinical presentation, eye examination, MRI and by measuring pituitary hormones. Treatment is with suppletion of the deficient hormones and in selected patients by transsphenoidal decompression surgery. PMID- 16375023 TI - [The significance of elevated troponin levels in the absence of acute cardiac ischaemia]. PMID- 16375024 TI - [Intracranial hypertension with severe and irreversible reduced acuity and impaired visual fields after oral tetracycline]. PMID- 16375025 TI - [Deliberate termination of life in newborns in The Netherlands; review of all 22 reported cases between 1997 and 2004]. PMID- 16375026 TI - The Bangkok Charter: impressions at its birth. PMID- 16375027 TI - The Bangkok Charter for health promotion in a globalized world. PMID- 16375028 TI - A charter to achieve health for all. PMID- 16375029 TI - The future of health promotion. PMID- 16375030 TI - Prolead: health promotion leadership development in the Western Pacific region. PMID- 16375031 TI - Is the term 'lifestyle' appropriate to use in health promotion today? AB - ISSUE ADDRESSED: The term 'lifestyle' has become popularised and is used across a variety of fields. This has diminished its currency in the health field somewhat. The term is also used to describe behaviour change and does not include social and environmental influences on health status. Many health professionals now question whether this term is appropriate to use in health promotion today. METHODS: A Google search on the world wide web produced 855,000 hits. A search for 'health lifestyle' in Australia alone yielded 23,000 hits. RESULTS: The Google search led to a variety of national and international reports related to lifestyle and healthy lifestyle as well as marketing reports such as the Australian Lifestyle Survey. The search helped to clarify the history of the term and its current use in health and other fields. CONCLUSIONS: There is no doubt the term 'lifestyle' has become popularised and is used to market everything from pets to health to pay TV. If the health field is to continue to use this term it needs to be prefixed with the word 'healthy'. However, it may be even more important for health professionals to be clear and consistent about what a 'healthy lifestyle' involves. PMID- 16375032 TI - Evaluation of an alternative transport initiative in Perth, Western Australia, 2000-04. AB - ISSUE ADDRESSED: Perth, Western Australia, has the highest proportion of per capita car ownership in Australia. Despite the various health, environmental and traffic-related benefits, the uptake of alternative forms of transportation such as walking, cycling and public transport are low. METHODS: In response to a growing reliance on motorised transport, the Department for Planning and Infrastructure's (DPI's) Cycling Unit conducts an annual Bike to Work Breakfast to promote cycling as a viable form of transport to work. During this event a short, cross-sectional survey is distributed to assess the usual cycling behaviours of attendees. Topical issues relating to cycling are also investigated. RESULTS: Results from the 2004 survey indicated that the majority of respondents cycled at least 2-3 times per week (85%) to improve fitness (88%), for enjoyment (70%) and because it was a cost-effective means of transport (45%). Most of the respondents (92%) supported the need for more cycle-friendly infrastructure such as advanced stop lines at intersections and safety measures such as wearing cycle helmets (85%). These findings were consistent with results from similar surveys conducted between 2000 and 2003. CONCLUSIONS: The DPI's Bike to Work Breakfast is a popular event within the cycling calendar that provides the opportunity to gain a snapshot of cycling behaviours and attitudes of cyclists. Prior consultation with the evaluators in the development of the survey instrument has the potential to investigate barriers and enablers to cycling on a regular basis. Age and gender differences could also be explored. PMID- 16375033 TI - Evaluating an indigenous sexual health peer education project. AB - ISSUE ADDRESSED: Many Indigenous Australians are dealing with a wide range of sexual health issues including contraception, HIV/AIDS and other sexually transmitted infections. The Indigenous Peer Education Program provided training to young, marginalised and disadvantaged Indigenous people to become sexual health peer educators within their communities. METHODS: The initiative comprised peer educator training, capacity building, and the development and dissemination of sexual health educational materials through an arts-based strategy. Evaluation involved a retrospective qualitative methodology, including document analysis, interviews and focus groups. RESULTS: The program was successful in training 22 young Indigenous peer educators and developing and disseminating more than 2,600 sexual health education resources to young Indigenous people and their community. CONCLUSIONS: The project had a positive immediate impact upon young Indigenous participants who reported and demonstrated increased knowledge of sexual health issues and skills as peer educators. The evaluation documented the program development and the immediate effects on participants, but did not enable an assessment of long-term effects for participants, service providers or the community. The evaluation identified areas for program improvement and challenges for program sustainability. The initiative was well supported within the local community and peer educators continued to engage in opportunistic sexual health promotion. PMID- 16375034 TI - A health impact assessment of an environmental management plan: the impacts on physical activity and social cohesion. AB - ISSUE ADDRESSED: This article describes a prospective health impact assessment (HIA) on a local government environmental management plan, the Shellharbour Foreshore Management Plan (SFM Plan). The health outcomes of interest were physical activity and social cohesion. METHODS: The procedures undertaken include: screening; scoping; assessment of potential health impacts; negotiation; and decision-making. The potential positive and negative health impacts of the SFM Plan were assessed and opportunities to enhance the positive and mitigate the negative were identified. RESULTS: Implementation of the SFM Plan was supported as the HIA showed a potential to benefit the health of local residents and visitors to the Shellharbour Foreshore area by increasing physical activity and social cohesion. CONCLUSION: The HIA provided a useful framework for establishing a collaborative relationship between the local government and health sector. It improved the local government's understanding of the inter-relationship between the environment and physical activity and social cohesion and enabled health authorities to have an impact on a capital works program to maximise health gain. PMID- 16375035 TI - A hidden curriculum: gambling and problem gambling among high school students in Auckland. AB - ISSUE ADDRESSED: Participation in gambling by young people aged 13-18 years. METHOD: During 2001, prior to the passing of legislation to minimise gambling harm, more than 500 students from six high schools completed a survey of their participation in gambling during the previous 12 months, and completed three problem gambling screens. RESULTS: Gambling, including under-age gambling, was a common event. Up to one in five were identified as at risk for problem gambling on at least one screen. Students who were non-European, or were from low socioeconomic areas, were more likely to be at risk for problem gambling. Help for gambling problems was preferred from friends and family rather than others, while inclusion of information in the education curriculum about risk of gambling problems was supported. CONCLUSIONS: The survey provided evidence for pre legislation baseline gambling behaviour, and risk for problem gambling, of students attending high schools in Auckland. Levels of risk for problem gambling paralleled the elevated risk found for youth in many countries. Raising awareness, through a school curriculum, of risk for gambling problems among adolescents may be explored as a strategy to reduce the high levels of risk for gambling problems identified. PMID- 16375036 TI - Improving health among culturally diverse sulbgroups: an exploration of trade offs and viewpoints annong a regional population health workforce. AB - ISSUES ADDRESSED: This article addresses availability and implications of evidence about culturally diverse subgroups in population. OBJECTIVES: To determine implications of evidence about culturally and linguistically diverse (CALD) communities for decision-makers and practitioners in population health. METHODS: Self-administered questionnaire of a regional population health workforce. One hundred and four population health staff of the Division of Population Health in South Western Sydney Area Health Service (SWSAHS) were invited to participate (response rate 73%). RESULTS: The majority of respondents were positive about applying culturally appropriate approaches in population health practice in south-western Sydney, a region well known for its cultural diversity. Respondents' levels of uncertainty increased when considering more strident statements, for example deciding exactly what to do when evidence generalisable to specific CALD communities was limited. CONCLUSIONS: Culturally appropriate approaches to population health are needed. As there is uncertainty in the workforce about how to deal with the paucity of ethno-specific evidence, a concerted effort is needed to study population-based interventions in order to increase the evidence base. PMID- 16375037 TI - Air pollution in Australia: review of costs, sources and potential solutions. AB - ISSUE ADDRESSED: Estimated health costs and principal sources of air pollution are reviewed, together with estimated costs of reducing pollution from major sources in Australia. METHOD: Emissions data from the Australian National Pollutant Inventory were compared with published estimates of pollution costs and converted to the cost per kilogram of emissions. Costs per kg of emissions (and, for the two main sources of pollution, diesel vehicles and wood heaters, costs per heater and per vehicle) are relatively easy to understand, making it easier to compare health costs with costs of pollution-control strategies. RESULTS: Estimated annual costs of morbidity/mortality exceed $1,100 per diesel vehicle and $2,000 per wood heater. Costs of avoiding emissions (about $2.1/kg PM2.5 for phasing out wood heaters and upwards of $70/kg for reducing diesel emissions) are considerably less than the estimated health costs ($166/kg) of those emissions. CONCLUSIONS: In other countries, smokeless zones (for domestic heating), heavy vehicle low-emission zones, and lower registration charges for low-emission vehicles reduce pollution and improve health. Similar 'polluter-pays' taxes in Australia to encourage retrofitting of existing diesels and incentives to choose new ones with lowest emissions would provide substantial benefits. Adopting Christchurch's policy of phasing out wood heaters and 'polluter-pays' levies to discourage their use would be extremely cost-effective. PMID- 16375038 TI - Trust levels of physical activity information sources: a population study. AB - ISSUE ADDRESSED: To examine levels of trust for various sources of physical activity information. METHODS: Questions from a 2003 nationally representative survey (n=8,291), Obstacles to Action, relating to how much respondents would trust various sources of physical activity information, were examined. RESULTS: The most trusted source of physical activity information was the general practitioner (GP) and the least trusted source was the Internet. High levels of trust were also found for health organisations, dietitians and nurses, while low levels of trust were also found for television, radio, and print media. CONCLUSION: The primary care setting continues to be the most trusted source of physical activity information. Those more likely to trust their physician were older adults, those with chronic disease, and the insufficiently active. PMID- 16375039 TI - A qualitative process evaluation of an oral health promotion program for older migrant adults. AB - ISSUES ADDRESSED: Qualitative process evaluation of a community-based health promotion program conducted to improve the use of oral health services, oral health knowledge, attitudes, and practices of older Greek and Italian adults living in Melbourne, Australia. METHOD: Fifteen focus groups were conducted with a sample of program participants (n=151) aged 55 and over, drawn from nine Italian and six Greek ethnic clubs. The focus groups sought participants' views about all aspects of the oral health promotion program and delivery, including the use of educational seminars; the preparation of simple language printed material known as the ORHIS (Oral Health Information Seminars/Sheets); and the distribution and demonstration of oral care products relevant to each individual seminar. RESULTS: Participants expressed positive views about the program and delivery approach. Findings indicated the importance of communicating dental information through a culturally relevant approach using easily understandable printed material in the participants' native languages; using the venue of their social clubs for health promotion efforts; and using an interactive and socially supportive approach that combined information giving, skills development and discussion in small groups over time. CONCLUSIONS: This process evaluation indicated that the ORHIS approach was perceived by participants as appropriate in assisting them to raise awareness of oral health issues and improving oral health knowledge and behaviours. This approach was highly acceptable to and valued by these communities. Findings suggest that when migrant populations are afforded greater consideration and time to learn about oral health concerns in an accessible and culturally relevant manner, the results may be quite positive. PMID- 16375040 TI - Does the type of promotional material influence men's attendance at skin screening clinics? AB - ISSUE ADDRESSED: The incidence of skin cancer is rising in many populations around the world. In Australia, men have both the highest incidence of thick melanoma and an increased risk of dying from this disease. Skin screening may provide a method for detecting lesions at an earlier stage. This study sought to assess the impact of two methods of encouraging men to attend free open-access skin screening clinics. METHODS: A randomised controlled trial of two recruitment methods. Subjects comprised all men aged 30 to 79 years (n=1,322) from a rural community. We randomly allocated 50% (661) to receive a personalised letter with the remaining 50% receiving the letter plus an additional brochure. RESULTS: Both groups were similar in age distribution. A total of 250 men (18.9%) attended the skin screening clinic. Overall, there was no difference in rates of attendance between the two groups. Younger men (30 to 49 years) in the letter and brochure group were more likely to attend a clinic, while men > or =50 years in the letter only group were more likely to attend (p=0.01) CONCLUSIONS: These results indicate that a personalised letter or targeted approach of invitation is effective in encouraging men to attend a skin screening clinic. The addition of health information through the use of a glossy brochure did not increase screening participation among men over 50. PMID- 16375042 TI - Making a difference. PMID- 16375041 TI - Tobacco smoking habits among Chinese medical students and their need for health promotion initiatives. PMID- 16375043 TI - Commissioning a patient-led NHS: implications for community practitioners and your clients. AB - The purpose of this Amicus Health Sector briefing is to provide an insight into the proposals set out in 'Commissioning a patient-led NHS'. We do not put forward an alternative blueprint. There will be discussion between Amicus members and oganisations that share our concerns. Emerging from this we expect a consensus view. Simply put, 'Commissioning a patient-led NHS' is at best a mechanism to address the continuing squeeze on primary care spending that is resulting in vacancy freezes and job cuts among care providers. At worst, it is the green light to creeping privatisation of primary care services PMID- 16375044 TI - Moving from reactive to proactive care: the Welsh Health agenda. PMID- 16375045 TI - Stigma over smear tests. PMID- 16375046 TI - A new primary care electronic record. What's in it for you? PMID- 16375047 TI - Sexual health promotion: the barriers school nurses face. AB - This paper reports on the findings of a study which aimed to determine the contribution of school nurses to promoting sexual health within schools and whether occupational and professional boundaries impinged on the school nurses' ability to undertake this aspect of their role. The research was carried out across three Primary Care Trusts (PCTs). Data were collected using semi structured interviews, a total of 30 school nurses (n=30) from across the three PCTs were included in the study. Analysis of the data indicated that barriers in the form of gate-keepers were present and they could impede the school nurse in fulfilling her role as a health educator. This gate-keeping existed across three 'tiers': the school governors, the school (as a collective organisation) and teachers. Nurses employed a variety of strategies to negotiate their way through these gatekeepers in order to access the classroom setting, key among them was their willingness to collaborate and co-operate, by 'slotting in' with the school, the curriculum and its timetable and 'fitting in' with the teachers, as their needs dictated. PMID- 16375048 TI - Service configuration and quality of Irish public health nursing. AB - This paper presents findings from a national study of the Irish public health nursing service and focuses, in particular, on issues relating to service configuration. The findings are drawn mainly from a national questionnaire of public health nurses (PHNs) working with families with infants (n=613; response rate 54 per cent) and the data were gathered in 1999/2000. The average ratio of public health nurse (PHN) to population size was found to be 1:3997 with a range between 500 and 16,500. The vast majority of respondents (85 per cent) had responsibility for five or more client groups including the elderly, those requiring clinical nursing care, terminal nursing care, psychiatric care, school nursing and the organisation of the home help service. Statistically significant differences between and within health board areas in the extent to which other nurses were available to the service led to a conclusion that in these circumstances a standardised service across individual PHN areas is not possible. The findings highlight the importance of the principle of vertical equity where service configuration is determined by population composition and need. PMID- 16375049 TI - When is a nurse not a nurse? Reflections on the NMC register. AB - Reform of the U.K. Nursing and Midwifery Council professional register has been undertaken through consultation over a period of some seven years from 1997, and in some aspects implementation remains ongoing. This paper reflects on the progress of the consultation and on its outcome: the provision of a professional register with three parts, the third part being designated for Registered Specialist Community Public Health Nurses. Confusion over the reasoning which led to this development is explored, in particular the status of health visiting as a profession distinct from nursing and midwifery and its place within the third part of the register. It is concluded that the arrangements for the third part which emanated from the consultation are contradictory and illogical. PMID- 16375050 TI - Predisposition to sensitive skin and atopic eczema. AB - Genes that control the thickness of our skin and its vulnerability to chemicals in the environment play a role in the development of contact dermatitis and atopic eczema. Sensitive skin manifests itself as a burning, stinging or itching sensation following the application of topical products such as soap, bubble baths and cosmetics. The skin may become red and dry after repeated application of these products. New insights into the skin barrier can help us improve treatment of the skin and prevent problems associated with atopic eczema and sensitive skin PMID- 16375051 TI - Agenda for change...the neverending story. PMID- 16375052 TI - Provision of care as and when. PMID- 16375053 TI - Early intervention in mental health. PMID- 16375054 TI - Medication in the wrong hands? PMID- 16375055 TI - Concern over bottle holding pillow. PMID- 16375056 TI - CPHVA MacQueen Award 2005. Leading by example. PMID- 16375057 TI - When the storms of life are raging. PMID- 16375058 TI - Orthopedic sports medicine: an abundance of resources. PMID- 16375059 TI - Management of midshaft clavicle fractures in adults. AB - Fractures of the clavicle are common injuries. The usual mechanism of clavicle fracture is a direct fall on the shoulder. There are 3 types of clavicle fractures, but type II or midshaft fractures make up the vast majority. Most clavicle fractures can be effectively treated nonoperatively. Rates of nonunion and poor functional outcome, however, may be higher than previously thought. Risk factors for nonunion include initial fracture displacement, comminution, shortening, and older age. The 2 main methods of operative management are plate and-screws and intramedullary fixation. Study results for both methods have been good. Indications for operative management, however, remain controversial. PMID- 16375060 TI - Autologous bone from hip or knee replacement in simultaneous foot or ankle arthrodeses. AB - Simultaneous operations on the lower extremities may potentially benefit some patients. However, the only literature discussing simultaneous operations has been limited to bilateral simultaneous total hip or knee replacements. The cases of 10 patients who had combined joint arthroplasty (8 knee, 2 hip) and foot and ankle arthrodesis (6 subtalar, 4 ankle) were reviewed retrospectively. Combining the procedures allowed the patient to have autologous bone brought from the joint replacement site to the foot or ankle to assist in filling defects in the arthrodesis. The fusion rate of 100% and the good results of hip or knee replacement demonstrate that successful treatment is possible when both conditions are treated simultaneously. PMID- 16375061 TI - Median nerve compression at the wrist caused by reversed 3-headed palmaris longus muscle: case report and review of the literature. PMID- 16375062 TI - Pronation-external rotation ankle fractures in 3 professional football players. AB - We found no reports of ankle fracture management in professional football players in the orthopedic literature. In this small series of ankle fractures in professional football players, all 3 had the same pronation-external rotation injury pattern. Given the high demands placed on the ankle by these professional athletes, we chose to use a 4.5-mm screw for fixation of the syndesmosis and a 3.5-mm reconstruction plate for fibular fixation to provide the most stable fixation. All 3 athletes were able to return to preinjury levels of athletic participation in the NFL without residual pain or stiffness. PMID- 16375063 TI - Athletes' ankle injuries: diagnosis and management. AB - Ankle injuries are common in the general and athletic populations. These injuries constitute 21% of all sports-related injuries. The wide spectrum of sports related ankle injuries includes ligamentous injuries, soft-tissue and osseous impingement, osteochondral lesions of the talus, tendon injuries, and fractures. Occult lesions (eg, fractures of the lateral process of the talus, fractures of the anterior process of the calcaneus, fractures of the base of the fifth metatarsal, os trigonum, stress fractures) may be missed on initial physical examination, and patients with such injuries often present to a sports clinic with persistent pain around the ankle. Because of increasing participation in sporting events, health care professionals involved in the care of athletes at all levels must have a thorough understanding of the anatomy, pathophysiology, and initial management of ankle injuries. In this review, we describe the pertinent anatomy, pathology, diagnosis, and treatment of sports-related injuries of the ankle. PMID- 16375064 TI - Early experience with high-deductible and consumer-driven health plans: findings from the EBRI/Commonwealth Fund Consumerism in Health Care Survey. PMID- 16375065 TI - Leading improvement. AB - PURPOSE: To provide research-informed guidance to leaders of quality and safety improvement and evidence-based materials for education programmes for leaders DESIGN/METHODOLOGY/APPROACH: Search of databases and hard copy literature since 1985 into managers' and leaders' role in quality and safety improvement. Classification into "eA": Empirical research strong evidence, "eB": Empirical research, weak evidence and, "eC": Conceptual discussion, not based on systematic empirical research. Summary, and synthesis of the best available evidence for a guidance checklist for leaders. FINDINGS: Although most literature emphasises the importance of committed leadership for successful quality and safety improvement, research evidence supporting this is scarce and often scientifically limited. The research shows evidence of the limitations and scope of leader actions for improving health care provision, the need to engage clinicians in this work and ways to do so, as well as the leadership role played by others apart from senior leaders. The ability of managers and other leaders skilfully to tailor Q&SI to the situation may be important but descriptions of how leaders do this and evidence supporting this proposition are lacking. RESEARCH LIMITATIONS/IMPLICATIONS: More research is needed about whether or how the leader role is different according to the stage of quality and safety development of the organisation, the type of organisation, the type of context, the level and type of leader and the type of improvement and improvement method. PRACTICAL IMPLICATIONS: Implications for leaders' actions are provided in a "best evidence guidance" checklist. This provides more tangible and research-informed guidance than the inspirational literature or studies from single organisations. ORIGINALITY/VALUE: This paper provides the first overview and synthesis of a wide range of studies which can be used as a basis for future research and materials for educational programmes. It provides the first detailed guidance for leaders about specific actions which research suggests they need to take to improve quality and safety. PMID- 16375066 TI - Cultural characteristics of "high" and "low" performing hospitals. AB - PURPOSE: To compare and contrast the cultural characteristics of "high" and "low" performing hospitals in the UK National Health Service (NHS). DESIGN/METHODOLOGY/APPROACH: A multiple case study design incorporating a purposeful sample of "low" and "high" performing acute hospital Trusts, as assessed by the star performance rating system. FINDINGS: These case studies suggest that "high" and "low" performing acute hospital organisations may be very different environments in which to work. Although each case possessed its own unique character, significant patternings were observed within cases grouped by performance to suggest considerable cultural divergence. The key points of divergence can be grouped under four main headings: leadership and management orientation; accountability and information systems; human resources policies; and relationships within the local health economy. PRACTICAL IMPLICATIONS: As with any study, interpretation of findings should be tempered with a degree of caution because of methodological considerations. First, there are the limitations of case study which proceeds on the basis of theoretical rather than quantitative generalisation. Second, organisational culture was assessed by exploring the views of middle and senior managers. While one should in no way suggest that such an approach can capture all important cultural characteristics of organisations, it is believed that it may be at least partially justified, given the agenda-setting powers and influence of the senior management team. Finally "star" performance measures are far from a perfect measure of organisational performance. Despite such reservations, the findings indicate that organisational culture is associated in a variety of non-trivial ways with the measured performance of hospital organisations. ORIGINALITY/VALUE: Highlights considerable cultural divergence within UK NHS hospitals. PMID- 16375067 TI - Instilling and distilling a reputation for institutional excellence: a critical reflection on organising practice. AB - PURPOSE: This ethnographic investigation of a general hospital aims to critically analyse a much lauded corporate culture. Rather than accepting the managerial and academic claims concerning the mobilisation of corporate culture at face value, this study builds upon a labour process analysis and takes a close look at how it actually seems to work. DESIGN/METHODOLOGY/APPROACH: The paper explores and describes how executive managers seek to design and impose corporate culture change and how it affects the nursing employees of this organisation. This was achieved by means of a six month field study of day-to-day life in the hospital's nursing division. FINDINGS: The results lend little support to the official claims that, if managerial objectives are realised, they are achieved through some combination of shared values and employee participation. The evidence lends more support to the critical view in labour process writing that modern cultural strategies lead to increased corporate control, greater employee subjection and extensive effort intensification. The contradiction this brings into the working lives of the employees leads to the conclusion that the rhetoric of corporate culture change does not affect the pre-existing attitudes and value orientations of nursing employees. However, there were considerable variations in how employees received the managerial message and thus, by their degree of misbehaviour and adaptation, affected the organisation itself as well as using the cultural rhetoric against the management for their own ends. ORIGINALITY/VALUE: The paper concludes that an extended labour process analysis is necessary to challenge the way in which corporate culture change is explored and described by management academics and practitioners. PMID- 16375068 TI - Using operational information and information systems to improve in-patient flow in hospitals. AB - PURPOSE: To consider how information and information systems can be used to support improving patient flow in acute hospitals (a key target for the National Health Service in England), and the potential role of the National Programme for Information Technology currently being developed. DESIGN/METHODOLOGY/APPROACH: The literature plus past and present research, teaching and consulting experience with all levels of the National Health Service is drawn on to consider information provision and requirements. FINDINGS: The National Programme for Information Technology specifies many features designed to support improving patient flows, though timescales for implementation are longer than those for the pledged flow improvements, and operational use of this type of information system has been problematic in the National Health Service. RESEARCH LIMITATIONS/IMPLICATIONS: The work is limited to the National Health Service and information systems in use and planned for it. The National Health Service access targets, flow improvement initiatives and the National Programme for Information Technology apply primarily to England. PRACTICAL IMPLICATIONS: Some bed/flow management information systems currently in use incorporate tools and capabilities in advance of what is outlined in the National Programme for Information Technology, and some rare cases of culture changes in information system use have been achieved. One should learn from these to inform development and implementation of National Programme systems. These existing information systems and approaches may also be useful to hospitals considering systems prior to implementation of the National Programme for Information Technology. ORIGINALITY/VALUE: There has been very little consideration of the use of operational information and information systems for bed/flow management in the literature. Development and implementation of National Programme for Information Technology systems should build from an understanding of the practice and context of bed/flow management. PMID- 16375069 TI - Women paediatricians: what made them choose their career? AB - PURPOSE: To explore why women paediatricians have chosen paediatrics as their specialty when previous research has shown that this was the least favourite career option. DESIGN/METHODOLOGY/APPROACH: In this qualitative study ten semi structured interviews with open-ended questions were carried out. The interviews were taped and transcribed by a professional transcriber. Two tapes could not be transcribed because of technical problems. This dissertation focuses on the responses of the eight women in relation to the research question. Interviews were carried out face-to-face. There were no additional questionnaires. Common themes were identified and content analysis used for data analysis. FINDINGS: All the women felt that they had chosen paediatrics because of the good work atmosphere, supportive colleagues and the attraction of working in a multi disciplinary team. All had pursued their career in a dedicated way working full time for most of their careers. The women paediatricians interviewed were happy with their career but they realised that they had made sacrifices in terms of family and personal commitments. The work environment has to change for everybody, not just for women, to improve quality of life. Career advice is still thought to be poor and many doctors enter a career without knowing what is expected of them. Modernising medical careers will considerably shorten medical training and the time individuals can spend in a particular specialty. ORIGINALITY/VALUE: A qualitative study like this can give both male and female students an insight into what it is like to have a career in paediatrics. PMID- 16375070 TI - Researchers' responses to research management and governance for primary care research in England: persistent and escalating problems over time. AB - PURPOSE: Research Governance has been introduced to regulate research involving National Health Service (NHS) patients and staff but the process is lengthy and bureaucratic and has improved little since its introduction. The paper seeks to investigate changes over time. DESIGN/METHODOLOGY/APPROACH: A comparison between two studies, one in 2003-2004 and the second in 2004-2005, provided the opportunity of observing the process of Research Governance as it was developing. Both studies included postal questionnaire surveys and interviews with study participants. FINDINGS: In order to achieve Research Governance approval, application forms have to be completed, reviews undertaken, sponsors and indemnity information identified and honorary contracts from individual PCTs applied for. Honorary contracts can require references and health and Criminal Records Bureau checks. The process reduces the time available for research and employs large numbers of people, yet is still inefficient. Some of the requirements reduce the quality of the research and researchers' rights are marginalised. RESEARCH LIMITATIONS/IMPLICATIONS: The studies took place in a rapidly changing research environment. Further studies should continue to assess the situation and report any progress in making Research Governance less onerous for researchers. PRACTICAL IMPLICATIONS: There is an urgent requirement to investigate how procedures can be simplified and made less bureaucratic and costly, and to protect the rights of researchers. Without this, primary care research will be seriously disadvantaged and may become impossible to pursue. ORIGINALITY/VALUE: The study compares process over time and shows that achieving Research Governance approval is seriously limiting research in primary care. PMID- 16375071 TI - Network structure and attitudes toward collaboration in a community partnership for diabetes control on the US-Mexican border. AB - PURPOSE: This study seeks to provide an examination of a health policy network operating in a single, small community along the US-Mexican border. The purpose of the paper is to discuss why and how this network evolved, and then to present findings on how the network was structured. Analysis will focus especially on agency involvement, or "embeddedness" in the network, and its relationship to attitudes held by network members regarding trust, reputation, and perceived benefit. DESIGN/METHODOLOGY/APPROACH: Data were collected from 15 public and nonprofit agencies trying to work collaboratively to influence local policy and services regarding the prevention of obesity-related chronic disease, especially diabetes. Embeddedness was measured in three different ways and both confirmed and unconfirmed networks were assessed. Network analysis methods were utilized as well as nonparametric correlation statistics. FINDINGS: The network was found to be densely connected through unconfirmed linkages, but much less so when these links were confirmed. Strongest findings were found for shared information. Measures of agency embeddedness in the network were strong predictors of agency reputation, but findings for trust and perceived benefit were generally weak. ORIGINALITY/VALUE: From a practice perspective, the study points to the problems in building and sustaining community-based chronic disease health networks, especially in a small community with substantial health needs. The research also contributes to theory on embeddedness and to methodology for collecting and analyzing data on community health networks. PMID- 16375072 TI - Project management and action research: two sides of the same coin? AB - PURPOSE: To put forward the, to date, unidentified viewpoint that organisational action research and project management have many shared properties--making it a useful exercise to compare and contrast them in relation to organisational management structures and strategies. DESIGN/METHODOLOGY/APPROACH: A conceptual exploration, drawing on a wide range of supporting literature, is used here. FINDINGS: Project management represents a mainstay strategy for much of the organisational research seen in health care management--and has done for many years. More recently, the exploratory literature on project management has identified many limitations--especially when matched against "traditional" examples. Many health services have witnessed a more recent organisational management drive to seek out alternative strategies that incorporate less hierarchical and more participatory research methods. Action research certainly fits this bill and, on further examination, can be incorporated into a project management ethos and vice versa. RESEARCH LIMITATIONS/IMPLICATIONS: The views expressed here are of a theoretical construct and have not been implemented, as they are presented in this paper, in practice. The intention, however, is to do so in some of the author's future studies. PRACTICAL IMPLICATIONS: If the management of health service organisations are to evolve to incorporate desirable structures that promote consumer-oriented empowerment and participation (where the consumers also include the workforce), then having a wider array of research tools at one's disposal is one way of facilitating this. Incorporating action research principles into project management approaches, or the other way round, or marrying them both to form a "hybrid" research strategy--it is argued here- represents an appropriate and representative way forward for future organisational management studies. ORIGINALITY/VALUE: In terms of originality, this represents a conceptual piece of work that puts forward constructs that have, to date, not featured in the health care literature. Its value lies in suggesting further options for organisational-oriented health care research. PMID- 16375076 TI - New directions? Forster report released. AB - Consultant Peter Forster handed over his final report on the Queensland Health Systems Review to the Premier and Health Minister on 30 September 2005. It was then immediately released to the public. The 491 page final report provides a broad overview of the reform needed in Queensland Health and details an initial three year reform programme. It also contains 388 recommendations including nursing specific recommendations. The QNU believes that the massive changes needed to deal with the problems identified in the report will require at least ten years to implement in full. The review took place over five months, received 1500 submissions including those from the QNU, its branches and individual members. PMID- 16375077 TI - Aged care. Dedication doesn't pay a nurses rent. PMID- 16375078 TI - WorkChoices--Critical complications for nurses' working lives. PMID- 16375081 TI - Doctors are heroes, too. PMID- 16375080 TI - "It's a heartache". PMID- 16375082 TI - 2005 TennCare survey validates physician concerns. PMID- 16375083 TI - Hurricane heroes. PMID- 16375084 TI - An all too frequent story. PMID- 16375085 TI - TMF growing pains require growing support. PMID- 16375086 TI - Don't forget year-end tax planning for 2005. PMID- 16375087 TI - Private wealth management requires personal attention. PMID- 16375088 TI - Public reporting needed to improve the health of Tennesseans. AB - Tennessee providers are recognizing an urgent need for a new and improved model of healthcare, characterized by transparent accountability to consumers. Meaningful health system improvements will require broad public disclosure of healthcare performance data at the hospital, clinic and community levels using nationally recognized standards. All Tennessee communities need a routine community health report card, to help their citizens to work together toward their most important health goals. PMID- 16375089 TI - Disclosing patient records containing HIV/STD information: a word to the wise healthcare practitioner. PMID- 16375090 TI - A new ethical dilemma: veterans with post-traumatic stress disorder. PMID- 16375091 TI - A Christmas Carol revisited--nursing's past, present, and future. PMID- 16375092 TI - The benefits of cholinesterase inhibitors: managing the behavioral and neuropsychiatric symptoms of Alzheimer's disease. AB - Pharmacological treatment can be helpful to improve cognition, functional ability, and behavior symptoms in older adults with Alzheimer's disease, resulting in reduced caregiver burden, delayed nursing home placement, and reduced health care costs. PMID- 16375093 TI - Studies of rural elderly individuals: merging critical ethnography with community based action research. AB - A model to help reduce health disparities in vulnerable populations is used in this study with Hispanic migrant farm workers in Colorado and multicultural rural older adults in southwestern New Mexico. PMID- 16375094 TI - Effectiveness of outpatient geriatric assessment programs: exploring caregiver needs, goals, and outcomes. AB - In this study, the authors examined the goals and outcomes of 141 caregivers of older adults with cognitive impairment who attended a comprehensive geriatric assessment program (CGA). The vast majority of caregivers expressed at least one assessment goal, suggesting that the concept was relevant to them. Most caregiver goals focused on patient needs, with only 10% expressing goals specifically related to their own coping needs. At follow up, more than two-thirds of caregiver goals were attained. The findings confirm the great potential of CGA programs in promoting positive outcomes and point to the opportunities for nurses to improve the process of care. PMID- 16375095 TI - Relaxing music at mealtime in nursing homes: effects on agitated patients with dementia. AB - Agitation in individuals with severe cognitive impairment is a significant problem that affects care and overall quality of life. Building on research conducted by Goddaer and Abraham (1994), this quasi-experimental study proposed that relaxing music played during meals would exert a calming effect and decrease agitated behaviors among nursing home residents with dementia. Thirty residents residing in a Special Care Unit participated in the 4-week study. The Cohen Mansfield Agitation Inventory (Cohen-Mansfield, Marx, & Rosenthal, 1989) was used to gather data. Baseline data was obtained in Week 1 (no music). Music was introduced in Week 2, removed in Week 3, and reintroduced in Week 4. At the end of the 4-week study, overall reductions in the cumulative incidence of total agitated behaviors were observed. Reductions in absolute numbers of agitated behaviors were achieved during the weeks with music and a distinct pattern was observed. PMID- 16375096 TI - Innovation in bladder assessment: use of technology in extended care. AB - Nursing homes are required to conduct a comprehensive assessment and screening of residents with urinary incontinence (UI) and indwelling catheter upon admission, and if there is a change in cognition, physical ability, or urinary tract function (Centers for Medicare and Medicaid Services [CMS], 2005). The desired goal is to improve the quality of care through the maintenance and the restoration of bladder function. CMS regulations and revised Tag F315 mandate that long-term care facilities appropriately assess and treat the specific bladder disorder of UI and decrease the inappropriate use of indwelling urinary catheters. Assessment includes evaluation for reversible factors that may cause UI, such as urinary retention and urinary tract infection. The scope of this article is to demonstrate the relationship between these bladder disorders and assessments that can be performed by nursing staff. Because invasive instrumentation of the bladder can increase the likelihood of urinary tract infections, the use of non-invasive technology that ensures quality and evidence based clinical practice, while preventing associated medical problems in vulnerable residents, is necessary for bladder assessment in the extended care setting. With this technology, assessment of bladder function is easy for staff and provides information that can be vital to successful nursing care outcomes. The staff of CRISTA Senior Community changed clinical practice through incorporation of technology to determine the presence of bladder dysfunction. Staff accurately and effectively applied ultrasonography to determine the bladder volume and abnormalities with bladder emptying, with ultimate improvement in resident care. Adoption of technology can preserve the resident's dignity and respect, while ensuring the highest level of bladder function. PMID- 16375097 TI - Enjoy! You thought coffee was bad for you? Actually, it seems to protect against all sorts of ills, from diabetes to liver cancer. PMID- 16375098 TI - Deficits draw calls for failure regime to get trusts on track. PMID- 16375099 TI - Their number's up--team deficit. PMID- 16375100 TI - Drug funding. Hewitt, Herceptin and the 100m pound sterling bill PCTs can't afford to pay. PMID- 16375101 TI - 'Direction of travel is robust.' Interview by Nick Edwards. PMID- 16375102 TI - Barometer. Acute trusts Nov 2005. PMID- 16375103 TI - Commissioning. Competition. Keep it local. AB - Local commissioners will require improved analytical skills to identify the specific health needs of their populations. Commissioning must be a counterweight to acutes that will be pushing for more work under contestability. PMID- 16375104 TI - Commissioning. Clinical engagement. Market forces. AB - Effective commissioning is a key aim of the forthcoming PCT reconfiguration. It is thought that many practices will initially opt out of commissioning. There are fears that the top-down NHS reorganisation will threaten effective commissioning. PMID- 16375106 TI - Case management. Matrons: are they really on the case? PMID- 16375105 TI - Commissioning. Skills. Out with the old. PMID- 16375107 TI - Good management. PMID- 16375108 TI - [Retrospective analysis of the diagnosis and surgical treatment of carotid body tumor]. AB - OBJECTIVE: To evaluate the diagnostic methods and the surgical treatment in carotid body tumor (CBT). METHOD: In retrospect of clinical data in 11 cases with CBT including CT, digital subtraction angiography (DSA), operation and its curative effect, the clinical experience in diagnosis and surgical treatment of CBT was summarized. RESULT: All 11 cases were classified as Shamblin I and II type. The cervical CT scan showed a tumorous lesion in the carotid bifurcation. DSA revealed definitely a solid vascular mass and its relation with the surrounding arteries. Preoperatively high selected embolization of the tumor- feeding artery could effectively reduce the intraoperative bleeding. All cases of CBT were resected without any carotid artery ligation. With 1-5 years follow-up, none of the CBT recurred in this 11 cases. CONCLUSION: CBT should be considered in patients with mass in the neck. The utilization of DSA could facilitate the diagnosis and the treatment of the CBT. Operation in the early stage of CBT could reduce the possibility of the injury of the arteries and the nerves. PMID- 16375109 TI - [Cat-scratch disease]. AB - OBJECTIVE: To study the etiology, epidemiology, clinical and pathological features, diagnose and treatment of cat-scratch disease(CSD). METHOD: The clinical information of 1 case was reported and the literatures were also reviewed. RESULT: Bartonella henselae was the primary pathogen of CSD. Almost all patients with CSD were in contact with animals, especially cat or dog before the occurrence of the disease. All the patients had the self-limited swelling of local lymph nodes and commonly a fever. The pathological feature was necrotic granulation-like micro abscessation, and a pleomorphous argyrophil G- bacillus could be showed by Warthin-Starry or Brown-Hopp stainings in the swelling lymph nodes. CONCLUSION: CSD is a self-limited bacterial infectious disease. And its clinical features,biopsy and special stainings of swelling lymph nodes are helpful to the final diagnosis. Gentamicin, rifadin, ciprofloxacin, clarithromycin, bactrim, mitramycin, or resection of swelling lymph nodes had good effects on treating CSD. PMID- 16375110 TI - [The effect of cervical circumference, lingual height and lingual width on the obstructive sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: To investigate the relation between the obstructive sleep apnea syndrome and the metrical value of the patient's cervical circumference, lingual height and lingual width. METHOD: The clinical and radiographic data of 33 (OSAHS) patients were retrospectively analyzed, the value of cervical circumference was direct manual measurement, the value of lingual height, lingual width was indirect measured through X-ray radiography of Iodic oil, and selected 33 normal adult as control group. RESULT: The mean of cervical circumference, lingual height and lingual width of OSAHS group have significant difference to normal control group respectively, the value of OSAHS group remarkably higher than control group. CONCLUSION: Cervical circumference, lingual height and lingual width make important effect on pathogenesis of OSAHS. PMID- 16375111 TI - [The observation of the ciliotoxicity of nasal mucosa with nasal decongestant]. AB - OBJECTIVE: To study the ciliotoxicity of nasalmucosal with nasal decongestant, in order to choose the appropriate nasal decongestant. METHOD: (1) Selected 100 healthy volunteers, by using 0.05% hydrochloride oxymetazoline, 0.025% hydrochloride oxymetazoline, 1.0% ephedrine and 0.5% ephedrine for 7 days with sacchariu technique to test nasal mucociliary transport, and compared with the normal saline. (2) To observe the mucosa of inferior turbinate with Electron Microscopy. RESULT: After using nasal decongestant for 7 days, Mean nasal mucociliary transport rate (MTR) in 0.0500 hydrochloride oxymetazoline groups was (7.64+/-1.56) mm/min and (7.46+/-1.65) mm/min in 0.25% hydrochloride oxymetazoline groups, Mean nasal mucociliary transport rate (MTR) in 1.0% ephedrine groups was (4.73+/-2.03) mm/min and (4.38+/-2.04) mm/min in 0.5% ephedrine groups,The normal saline groups was (7.14+/-1.76) mm/min. MTR was obviously different (F = 16.50, P <0.01). (2)With electron microscopy, it was found that the cilia of epithelial cells were in good order, uniformed in hydrochloride oxymetazoline groups and exfoliated in ephedrine groups. CONCLUSION: Ephedrine has ill effects on nasal mucosa; Hydrochloride oxymetazoline has neither effect on nasal mucosa, nor destroying the physical function of nose, it is an ideal nasal decongestant. PMID- 16375112 TI - [Significance of the changes of urinary uric acid in OSAHS before and after UPPP]. AB - OBJECTIVE: To assess the value of urinary uric acid excretion and urinary uric acid/creatinine ratio as the marker of obstructive sleep apnea-hypopnea syndrome (OSAHS) before and after Uvulopalatopharyngoplasty (UPPP). METHOD: Twenty three cases diagnosed as OSAHS by polysomnography (PSG) were treated as trial group, 12 cases excluded from OSAHS by PSG were taken as control group,and 15 patients of the trial group were treated by UPPP and were taken as before UPPP therapy group and after UPPP therapy group. The makers above were compared in these groups. RESULT: The overnight change in urinary uric acid/creatinine ratio in trial group is 0.46+/-0.25, and after UPPP therapy group (0.13+/-0.2) significantly lower than before UPPP therapy group (0.49 +/-0.32), P < 0.01. CONCLUSION: The urinary uric acid excretion and overnight change in urinary uric acid/creatinine are good markers to determine the effects of UPPP on OSAHS. PMID- 16375113 TI - [Long term follow up of laser surgery under micro-endoscopy for laryngeal carcinoma]. AB - OBJECTIVE: To observe the long term curative effect for laryngeal carcinoma treated with CO2 laser under micro-endoscopy. METHOD: Twenty cases of laryngeal cancer were treated by CO2 laser under micro-endoscopy. All patients were followed up at least 5 (5-8) years by phone, letters or clinical review. RESULT: During 5 to 8 years following up, one patient died of recurrence 1 year later. Another was operated on with partial laryngectomy due to local recurrence 2 years after laser surgery. The third one was operated with selective neck dissection because of neck metastasis 5 years later. Other 17 cases were alive without recurrent signs. The 5 years total cure rate was 85% and survival rate was 95%, the 5 years local recurrence rate was 10% and the neck metastasis rate was 5%. All survivals had normal breath and good phonation. Among them 8 resumed their daily works, 5 were used to do house works and 4 did them sometimes. CONCLUSION: T1 and earlier glottic or supraglottic cancers are indications of laser surgery under micro-endoscopy. Its 5 years survival rate as well as live quality were satisfactory and proved the method to be a choice of treatment for earlier laryngeal carcinoma. PMID- 16375114 TI - [Compare the differences between automatic analysis and manual analysis of apnea events in polysomnogram]. AB - OBJECTIVE: To investigate whether there are differences on the diagnose of sleep apnea hypopnea syndrome between automatic analysis and manual analysis of respiration events in polysomnogram. METHOD: Sixty snoring human was selected randomly and monitored separately by sleep monitor in seven hour at night, sleep monitor would produce a poly sonogram, firstly polysomnogram was analysis by the software of sleep monitor and then by the manual operation. Finally data were processed by statistical analysis. RESULT: There were significant differences between automatic analysis and manual analysis of polysomnogram on apnea hypopnea index,apnea index, hypopnea index and long apnea. Data of manual analysis were more consistent with clinic symptoms of patients than automatic analysis. CONCLUSION: The result of automatic analysis of polysomnogram had a deviation to the diagnosis of pathogenetic, in order to improve the accuracy of clinic diagnosis, polysomnogram should be analysis by manual operation generally. PMID- 16375115 TI - [Expression and biological significance of survivin and p63 in laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To investigate the expression of Survivin and p63 and their biological significance in laryngeal squamous cell carcinomas (LSCC). METHOD: The expression of Survivin and p63 in 47 cases of tumor tissues, adjacent and normal tissues were detected using in situ hybridization. RESULT: Percentage of Survivin expression was 59.6% (28/47), 25.5% (12/47) in tumor tissues and adjacent tissues respectively. In contrast, normal laryngeal tissues did not express Survivin. Over expression of Survivin was strongly associated with tumor grade, TNM stage and cervical nodal metastasis. Percentage of p63 expression was 91.5% (43/47), 93.6% (44/47), 57.1% (8/14) in tumor tissues, adjacent and normal tissues respectively. The expression of p63 was not related to any clinicopathological features. CONCLUSION: Survivin may play an important role in the pathway of carcinogenesis and progression of LSCC and it may be identified as a new therapeutic target. The up-regulation of the expression of Survivin predicts more invasion and poor prognosis. p63 has less effect on biological behaviors of LSCC. PMID- 16375116 TI - [Clinical research of artificial larynx]. AB - OBJECTIVE: Using delayed degradation bone matrix gelatin (BMG) to repair the defect of laryngeal frame for the goal of laryngeal external form and function recovery. METHOD: Partial laryngectomy and total laryngectomy were performed in nine cases with laryngeal cancer, then delayed degradation BMG be implanted in the defect of laryngeal frame or inside sterno hyoid muscle to build the laryngeal external form. In the same time the delayed degradation BMG be implanted in the crico-arytenoid muscle and around fibre with nerve ending, then embedding and sewing up to recover the swallow function and to prevent deglutition disorder. RESULT: The delayed degradation BMG were applied in nine cases to rebuild new larynx, five of them were performed vertical partial laryngectomy and two total laryngectomy. The effect is satisfied after 15 years followed up. Eight of them rebuild the laryngeal external form and function, one patient need put on tube for long-term and cramming to speaking. CONCLUSION: Applying delayed degradation BMG to repair the laryngeal defect not only rebuild the laryngeal external form, but also recover the laryngeal function. It is an ideal bioactivity material and best choice for the new larynx rebuilding in the operation now. PMID- 16375117 TI - [The expressions of survivin and VEGF in squamous cell carcinoma of larynx and the correlation between the two marks]. AB - OBJECTIVE: To study the expressions and biological behavior of Survivin and VEGF in squamous cell carcinoma of larynx (LSCC) and the correlation between Survivin and VEGF. METHOD: The expressions of Survivin and VEGF in LSCC tissues of 40 cases and normal mucosa of larynx tissues of 5 cases were studied by immunohistochemical staining. RESULT: The expressions of Survivin and VEGF in LSCC tissues were higher than those in normal mucosa of laryngeal cavity (P <0.01). The expressions of the two marks in LSCC with the metastasis of lymph nodes were higher than those without metastasis (Survivin: P <0.05, VEGF: P <0.01). Age and sex did not change the expressions obviously (P > 0.05). There was a negative correlation between T-stage and Survivin expressions (P < 0.05). There was also a negative correlation between T-stage and VEGF expressions (P < 0.05). But, there was a positive correlation in staining intensity between Survivin and VEGF (r = 0.676, P < 0.01). CONCLUSION: Significant correlation between Survivin/VEGF expression and T-stage (P <0.05), lymph node metastasis (P <0.05) and prognosis (P <0.01) was revealed in squamous cell carcinoma of larynx. The high expression of it suggests bad prognosis. The up-regulate of the two genes suggest that they play important roles in the formation and metastasis of LSCC. PMID- 16375118 TI - [Expression of vascular endothelial growth factor and its receptors in laryngeal carcinoma cell and its significance]. AB - OBJECTIVE: To study the expression of vascular endothelial growth factor (VEGF) and both the Flt-1 and KDR high affinity VEGF receptors in human laryngeal carcinoma cells and its significance. METHOD: In this study, we investigated the expression of VEGF mRNA and both the Fit-1 mRNA and KDR mRNA high affinity VEGF receptors in human laryngeal carcinoma cells using techniques of semi-quantify RT PCR. RESULT: It was showed that the expression level of VEGF mRNA was significantly increased in human laryngeal carcinoma cells as compared with the normal cells, Hacat cells (P <0.05), and there was the high expression level of Flt-1 mRNA in the Hep-2 cells, but not KDR mRNA. CONCLUSION: These data raise the possibility that VEGF and its receptors many play key roles in the growth, invasion and metastasis of human laryngeal carcinoma cells through either a paracrine or an autocrine mechanism. PMID- 16375119 TI - [The expression and role of VEGF-C and its receptor FLT-4 in laryngeal and hypopharyngeal squamous cell carcinoma metastasis]. AB - OBJECTIVE: To investigate the expression of VEGF-C and its receptor FLT-4 in laryngeal and hypopharyngeal squamous cell carcinoma, and to analyse its role in lymphatic metastasis and micrometastasis. METHOD: Forty nine cases postoperative specimens verified by histopathology were studied. The expression of VEGF-C, FLT 4 were detected with immunohistochemical SP method, image analysis method. The clinicopathological parameters of tumor were compared with the immunohistochemical results of VEGF-C and FLT-4, followed by the further analysis on the relationship between VEGF-C/FLT-4 expression and lymph nodal metastasis or micrometastasis. RESULT: There was a statistical correlation between the expression of VEGF-C and grades of pathological differentiation in laryngeal and hypopharyngeal squamous cell carcinoma. Moreover there was also a statistical correlation between the expression of VEGF-C and lymph node metastases. But no correlation was observed between the expression of VEGF-C and clinical T grades. The statistical correlation also existed between VEGF-C and FLT-4 expression in laryngeal or hypopharyngeal tumors. There was a significant difference of expression of FLT-4 in laryngeal and hypopharyngeal tumours with positive and negative lymph node metastases. No correlation was observed between FLT-4 and grades of pathological differentiation or clinical T grades. There was no statistical correlation between the expression of VEGF-C/FLT-4 and nodal micrometastasis (P > 0.05). CONCLUSION: VEGF-C and its receptor FLT-4 play an important role in the lymphatic metastasis of laryngeal and hypopharyngeal squamous cell carcinoma. VEGF-C/FLT-4 may be a useful biological marker to foresee the cervical lymph node metastasis and prognosis. PMID- 16375120 TI - Monkey see, monkey do. PMID- 16375121 TI - The 'hunger season'. PMID- 16375122 TI - Reading your baby's mind. PMID- 16375123 TI - America's most dangerous drug. PMID- 16375124 TI - How to fix school lunch. PMID- 16375125 TI - How to quit the cure. PMID- 16375126 TI - Small is beautiful. PMID- 16375127 TI - A dream before dying. PMID- 16375128 TI - A 9,000-year-old secret. PMID- 16375129 TI - Babies who take Viagra. PMID- 16375130 TI - What we owe what we eat. PMID- 16375131 TI - Your health in the 21st century. The future of medicine. PMID- 16375132 TI - Your health in the 21st century. Making chemo easier to take. PMID- 16375133 TI - Your health in the 21st century. Do stem cells cause cancer? PMID- 16375134 TI - Your health in the 21st century. The next revolution. PMID- 16375135 TI - Your health in the 21st century. Fine-tuning heart health. PMID- 16375136 TI - Your health in the 21st century. A race-based heart remedy. PMID- 16375137 TI - Your health in the 21st century. Quieting a body's defenses. PMID- 16375138 TI - Your health in the 21st century. The best ways to beat diabetes. PMID- 16375139 TI - Your health in the 21st century. Keeping HIV at bay. PMID- 16375140 TI - Your health in the 21st century. 7 ways to save a brain. PMID- 16375141 TI - Your health in the 21st century. Managing every shade of blue. PMID- 16375142 TI - Your health in the 21st century. Building better medicines. PMID- 16375143 TI - Your health in the 21st century. The shape of things to come. PMID- 16375144 TI - Your health in the 21st century. Organs under construction. PMID- 16375145 TI - Your health in the 21st century. It's a gene pool party. PMID- 16375146 TI - Your health in the 21st century. Silencing bad genes. PMID- 16375147 TI - Your health in the 21st century. A fix for faulty proteins. PMID- 16375148 TI - Your health in the 21st century. Pharmacy island. PMID- 16375149 TI - Your health in the 21st century. Chasing black fever. PMID- 16375150 TI - Your health in the 21st century. On the march to eradicate child illness. PMID- 16375152 TI - Your health in the 21st century. A better safety net. PMID- 16375151 TI - Your health in the 21st century. A new threat in the lab. PMID- 16375153 TI - Your health in the 21st century. Getting all the facts on drugs. PMID- 16375154 TI - Your health in the 21st century. An Rx for kids--with warnings. PMID- 16375155 TI - Your health in the 21st century. Progress in preventing errors. PMID- 16375156 TI - Your health in the 21st century. Medicine tailored just for you. PMID- 16375158 TI - Your health in the 21st century. Get ready to get wired. PMID- 16375157 TI - Your health in the 21st century. The right time for a cure. PMID- 16375159 TI - Your health in the 21st century. The frontier: interventional radiology. PMID- 16375160 TI - Technology's limits. PMID- 16375161 TI - Health for life. Cutting edge. PMID- 16375162 TI - Health for life. Immunity's master controller. PMID- 16375163 TI - Health for life. Genomic medicine: the science speeds up. PMID- 16375164 TI - Health for life. Addiction: how to break the chain. PMID- 16375165 TI - Health for life. The ultimate transplant. PMID- 16375166 TI - Health for life. The man who lost his face. PMID- 16375167 TI - Health for life. A teen health gap. PMID- 16375168 TI - Health for life. The cost of the Katrina effect. PMID- 16375169 TI - Health for life. Bracing for a plague. PMID- 16375170 TI - Health for life. 6 keys to safer hospitals. PMID- 16375171 TI - Health for life. Diagnosis: not enough nurses. PMID- 16375172 TI - Health for life. Beyond the horizon. PMID- 16375173 TI - Tracking disease. PMID- 16375174 TI - Sweetness and fight. PMID- 16375176 TI - You are what you eat. PMID- 16375175 TI - The fight against the flu. PMID- 16375177 TI - The race against avian flu. PMID- 16375179 TI - Nature's design workshop. PMID- 16375178 TI - A problem in the brain. PMID- 16375180 TI - Some are found all are lost. PMID- 16375181 TI - Boy brains, girl brains. PMID- 16375182 TI - Critical condition. PMID- 16375183 TI - An unexpected reprieve. PMID- 16375184 TI - The deadliest cancer. PMID- 16375185 TI - Peter Jennings. PMID- 16375186 TI - School design for the blind: learning to see without sight. PMID- 16375187 TI - [Treatment for malignant glioma]. PMID- 16375188 TI - [Treatment for low grade glioma focusing on grade II glioma]. PMID- 16375189 TI - [Treatment of central nervous system malignant lymphoma]. PMID- 16375190 TI - [Treatment of central nervous system germ cell tumors]. PMID- 16375191 TI - [Recent management of metastatic brain tumors]. PMID- 16375192 TI - [Hippocampal atrophy and memory disturbance]. PMID- 16375193 TI - [A case of pseudo Tolosa-Hunt syndrome with bacterial infection and literature review]. AB - We report a case with recurrent orbital pain and unilateral cranial nerve paresis mimicking Tolosa-Hunt syndrome. However, these features were most likely caused by bacterial infection because of beneficial response to antibiotics therapy. A 32-year-old man developed severe right orbital pain and diplopia. Neurological examination revealed right oculomotor paresis and 1st division of the right trigeminal nerve dysfunction. MR imaging revealed thickness of right cavernous sinus region with marked gadolinium enhancement. Cerebrospinal fluid (CSF) examination was initially normal. Treatment with steroid showed marked improvement. However soon after tapering of steroid dosage, his symptoms recurred and deteriorated. He was referred to our hospital because of second opinion. Neurological examination still showed right oculomotor paresis and 1st division of the right trigeminal nerve dysfunction with orbital and retro-orbital pain. Re examination of CSF showed pleocytosis with neutrocytes dominancy and elevated protein concentration. Intravenous treatment with penicillin was initiated with marked improvement. There have been reported cases with bacterial infection resulting pseudo Tolosa-Hunt syndrome, which have good response to antibiotics treatment with excellent prognosis. It is speculated that bacterial infection might cause clinical features mimicking Tolosa-Hunt syndrome in our case. PMID- 16375194 TI - [Cerebral small infarcts of the precentral and postcentral gyri presenting with unilateral monoataxia: a report of two cases]. AB - Two cases of unilateral monoataxia, due to small infarcts in the precentral and postcentral gyri, were reported. A 76-year-old man (case 1), and a 90-year-old woman (case 2) suddenly developed clumsiness of the left upper extremity. Neurological examination revealed cerebellar ataxia in the left upper extremity in both cases. But, the other abnormal neurological findings, including the muscle weakness, abnormal muscle tone, and proprioceptive deficit, were not noted. In case 2, cerebellar ataxia disappeared about ten days after the onset of the disease. On the other hand, ataxia of case 1 remained about two months after the disease onset. Brain magnetic resonance imaging (MRI) of cases 1 and 2, showed small infarcts at the border between the right precentral gyrus and postcentral gyrus. In addition, brain MRI of case 1 disclosed another infarcts in the just medial portion of the precentral knob and the centrum semiovale of central region, respectively. It was suggested that the mechanism of cerebellar ataxia caused by infarct in the central region, was not only due to the interruption of two distinctive neuronal pathways, including the corticopontine tract and cerebellothalamocortical tract, but also due to the disturbance of sensory-motor integrity. In conclusion, etiologies of unilateral monoataxia may be heterogeneous. Furthermore, functional outcome may depend on the mechanism of unilateral monoataxia. PMID- 16375195 TI - [A case of acute cervical epidural abscess]. AB - Spinal epidural abscess (SEA) is a rare condition with potentially devastating consequences and more aggressive diseases than previously recognized. We report a case with cervical epidural abscess treated successfully by emergent neural decompression. A 49-year-old man presented with fever and cervical pain. An magnetic resonance imaging (MRI) showed an epidural abscess at C5-6. His condition deteriorated with the development of complete sensory loss and tetraparesis. Emergent anterior neural decompression was performed, and pus was aspirated which grew Staphylococcus aureus. Adequate antibiotics were administered, and the patient showed remarkable improvement of neurological signs and symptoms. He underwent reconstruction surgery 3 weeks after the first operation, and returned to his original job 3 months after the operation. Prompt diagnosis and satisfactory surgical decompression are essential to avoid irreversible neurological sequelae in epidural abscess of the cervical spine. PMID- 16375196 TI - [Case report of spontaneous cervical epidural hematoma in elderly]. AB - Spontaneous spinal epidural hematomas (SSEH) are relatively rare clinical entities and associated with coagulopathies, tumors, or vascular malformation. In addition, these are often neurosurgical emergencies; therefore, prompt diagnosis and treatment are paramount. We reported a case of an 87-year-old woman with spontaneous cervical epidural hematoma. She presented with the sudden onset of neck pain, rt. upper arm sensory disturbance and rt. hemiparesis. MRI revealed a C3-Th1 dorsally placed extradural lesion. The lesion was iso-to hypointense on T1 weighted images and was hyperintense on T2-weighted images. She underwent surgery within 9 hours after symptom onset, removal of hematoma through the right hemilaminectomy was performed. She was a very high age, but she was discharged from the hospital without neurological deficit. To obtain good prognosis for patients with SSEH, early diagnosis and treatment are important. We also review the current literature concerning diagnosis and treatment of SSEH. PMID- 16375197 TI - [Magnetic resonance images of traumatic MLF syndrome]. PMID- 16375199 TI - Beta2 agonist reversibility testing in COPD diagnosis. AB - The place of testingthe reversibility of airway obstruction with an inhaled short acting beta2 agonist in the diagnosis of chronic obstructive pulmonary disease (COPD) has caused some confusion in primary care. When reviewing the management of patients with stable COPD, in 2001, we stated that "to help rule out asthma, reversibility testing should be undertaken by measuring FEV1 [forced expiratory volume in 1 second] before and after the patient inhales a short-acting beta2 agonist". In keeping with this idea, the current general medical services (GMS) contract for GPs in the U.K. requires confirmation of the diagnosis of COPD with both spirometric evidence of airflow obstruction and reversibility testing of such obstruction. Other national and international guidelines have also recommended this approach. However, the National Institutefor Health and Clinical Excellence (NICE) guideline on COPD statesthat, in most patients, routine spirometric reversibility testing is not necessary in the diagnostic process, and that it should only be used when there is diagnostic uncertainty or if the patient is thought to have both COPD and asthma. Here we reconsider the role of reversibility testing with a short-acting beta2 agonist in the diagnosis of COPD. PMID- 16375198 TI - [A 54-year-old man with familial parkinsonism, gaze palsy, and dementia]. AB - We report a Japanese man with familial parkinsonism who died at age 54. His younger brother, his mother, the mother's 4 brothers, and their mother were also affected with similar parkinsonism. The patient had had nystagmus since adolescence. He noticed difficulty in walking and micrographia at age 42. Neurological examination at age 45 in our hospital revealed pendular nystagmus, moderate rigidity in his neck and upper limbs, postural tremor in hands and shuffling gait. He received L-dopa/benzerazide 200 mg and his movement was mildly improved. Then he developed forced closing of eyelids suggesting either blepharospasms or apraxia of eye lid opening. He became apathetic at age 48. He was admitted to our hospital at age 49. On admission, he showed mild dementia and sexually disinhibited behaviours. Moderate downward gaze palsy and rigidity were seen. Increase of L-dopa/benzerazide and pergolide did not improve his parkinsonism and his disinhibited behaviors became worse. L-dopa/benzerazide and pergolide were decreased and he received electroconvulsive therapy at a psychiatric hospital with temporally improvement in his movement. He became unable to walk at age 52 and he was mutic and bedridden. He died of pneumonia at age 54. The patient was discussed in a neurological CPC, and a chief discussant arrived at the conclusion that the patient had a familial form of dementia with Lewy bodies. Many participants thought that he had frontotemporal dementia and parkinsonism linked to chromosome 17. The pathological examination of his brain showed severe neuronal loss in the substantia nigra, subthalamus, and pallidum. Ballooned neurons were observed in the cerebral cortex. Immunohistochemistry using anti-tau antibodies revealed tau-positive neurons, glial cells and threads in the cerebral cortex, white matter and subcortical nuclei; these tau deposition reacted with an anti-4-repeat tau antibody, but not reacted to an anti-3-repeat tau antibody. Sequencing of genomic DNA of the patient showed a missense mutation in exon 10 of tau that caused a substitution at codon N279K. These neuropathological and molecular studies revealed the diagnosis of the patient was FTDP-17 with N279K mutation. PMID- 16375200 TI - Action plans in asthma. AB - Around 5.2 million people in the U.K. have asthma. Symptoms of the disease are often variable, so a fixed treatment regimen is not always appropriate. Personal asthma action plans aim to prompt patients to adjust their medication, or seek medical help, in response to changes in symptoms or peak flow readings. A national clinical guideline in the U.K. recommends that patients with asthma should be offered education in self-management, including a written asthma action plan that focuses on their individual needs. At present, only a minority of people with asthma report receiving such an action plan. Here we discuss the potential content of action plans and review the evidence for their effectiveness. PMID- 16375201 TI - Chickenpox, pregnancy and the newborn: a follow-up. AB - In September 2005, we published an article on Chickenpox, pregnancy and the newborn. One of the issues it considered was fetal varicella syndrome, an uncommon but potentially fatal consequence of in-utero chickenpox infection. We reported evidence suggesting that contracting maternal chickenpox within the first 28 weeks of pregnancy can lead to fetal varicella syndrome. This suggestion was questioned after publication since it was at odds with national guidance in the U.K. and elsewhere, which has indicated that fetal varicella syndrome occurs only where maternal chickenpox develops before 20 weeks of pregnancy. Here we discuss in more detail the basis for our conclusion and its implications. PMID- 16375202 TI - [Visual acuty after trabeculectomy]. AB - BACKGROUND/AIM: In this study we compared the two groups of glaucoma patients who had underwent trabeculectomy, with or without intraoperative application of antimetabolite 5-Fluorouracile (5 FU). METHODS: We followed up 50 glaucoma patients (group I) in a four-years period after trabeculectomy alone (antimetabolites were not applied), and 50 patients in whom antimetabolite 5 FU was applied (group II). We followed up intraocular pressure (IOP), visual acuity (VA), lens transparency, optic disc changes, visual fields (VF perimetry). The IOP at the time of the surgery, and the period of the treatment before the surgery were the important data for the follow-up. RESULTS: The results that we got in the first group were: in 8 patients (16%) there was no change of VA, and of optic disc with a mild VF deterioration; in 18 patients (33%) the worsening of VA by 1 Snellen line (3 years after the surgery) with no change of the disc and a mild VF progressing; in 10 patients (20%) VA decreased by 2 Snellen lines with the progressive VF changes and cataract appearance, (cataract and VA worsening appeared mainly 3 years after the surgery); in 6 patients (12%) by 3 Snellen lines, and in 4 patients (8%) by 4 Snellen lines. In 4 patients (8%), the highest worsening of VA--(0.2) and the best (0.9) at the time of the surgery were observed. The VA ranging from 0.4-0.6 at the time of the surgery yielded the longest preservation of vision. In the group II (5 FU applied intra-operatively): in 5 patients (10%) there was no change of VA and of optic disc with a mild VF deterioration; in 20 patients (40%) the worsening of VA by 1 Snellen line, with no change of the disc and a mild VF progressing. In 14 patients (28%) VA decreased by 2 Snellen lines with the progressive VF changes and cataract appearance; in 7 patients (14%) by 3 Snellen lines, and in 4 patints (8%) by 4 Snellen lines. CONCLUSION: All of the checked parameters were better in the group II of the patients, (5 FU applied intraoperatively). Also, in the group of the patients with the longest treatment before the surgery and with the good VA, and in the patients with the shorter treatment and with the bad VA, the quickest progression of the vision losts was observed. The best preservation of VA was recordered in the patients with a mild IOP, and in those who had not been treated more than 1 or 2 years with the drops before the surgery. The VA ranging from 0.4 0.6 at the time of the surgery allowed the longest preservation of vision. PMID- 16375203 TI - [Influence of orlistat therapy on serum insulin level and morphological and functional parameters of peripheral arterial circulation in obese patients]. AB - BACKGROUND/AIM: Insulin resistance is related to accelerated atherosclerosis, whereas weight loss is associated with the increasing insulin sensitivity, the improvement of functional and the morphological parameters of arterial circulation, and the reduction of cardiovascular morbidity and mortality. The aim of our study was to evaluate the influence of orlistat treatment on serum insulin level and functional and morphologic parameters of peripheral arterial circulation. METHODS: We conducted a prospective, randomized, double - blind, placebo - controlled study. Thirty patients with body mass index over 30 kg/m2 normotensive, nonsmokers, without clinically manifested cardiovascular disease or diabetes were randomly assigned either orlistat (120 mg, 3 times daily; n = 20) or placebo (n = 10) in a double - blind manner. All of the patients were on individually calculated hypocaloric diet. The follow-up period was 24 weeks. Arterial pressure, fasting serum glucose and insulin level, triglycerides, total cholesterol and low density lipoprotein-cholesterol were determined at the beginning, following 3 and 6 months. Also, the intima - media thickness of right superficial femoral artery and the mean blood flow velocity were determined with ultrasonography. RESULTS: Inside the period of 3 and 6 months, there were the greater reductions of body mass index, arterial pressure, fasting glucose and insulin level, total cholesterol, low density lipoproteins, as well as the greater reductions of mean velocity blood flow and peripheral pulse pressure in the orlistat group vs the placebo group (p < 0.01). Greater reductions in the waist circumference and intima - media thickness were registered following 6 months in the orlistat vs the placebo group (p < 0.01). CONCLUSION: In the group of obese patients orlistat therapy reduced risk factors, serum insulin level and improved early arterial functional changes as assessed with the reductions of the mean velocity blood flow and peripheral pulse pressure following 3 months. The regression of morphological changes, as assessed with the reduction in intima - media thickness, was feasible over the six - month period. PMID- 16375204 TI - [Relationship between C-reactive protein and features of the metabolic syndrome in military pilots in the Serbia and Montenegro]. AB - BACKGROUND/AIM: C-reactive protein is an independent predictor of the risk of cardiovascular events and diabetes mellitus in apparently healthy men. The relationship between C-reactive protein and the features of metabolic syndrome has not been fully elucidated. To assess the cross-sectional relationship between C-reactive protein and the features of metabolic syndrome in healthy people. METHODS: We studied 161 military pilots (agee, 40 +/- 6 years) free of cardiovascular disease, diabetes mellitus and active inflammation on their regular annual medical control. Age, total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol, triglycerides, fasting glucose, glycosylated hemoglobin, blood pressure, smoking habit, waist circumference and body mass index were evaluated. Plasma C-reactive protein was measured by the immunonephelometry (Dade Behring) method. Metabolic syndrome was defined according to the National Cholesterol Education Program Expert Panel. RESULTS: The mean C-reactive protein concentrations in the subjects grouped according to the presence of 0, 1, 2 and 3 or more features of the metabolic syndrome were 1.11, 1.89, 1.72 and 2.22 mg/L, respectively (p = 0.023) with a statistically, significant difference between those with 3, and without metabolic syndrome (p = 0.01). In the simple regression analyses C-reactive protein did not correlate with the total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol, body mass index and blood pressure (p > 0.05). In the multiple regression analysis, waist circumference (beta = 0.411, p = 0.000), triglycerides to high density lipoprotein cholesterol ratio (beta = 0.774, p = 0.000), smoking habit (beta = 0.236, p = 0.003) and triglycerides (beta = 0.471, p = 0.027) were independent predictors of C-reactive protein. CONCLUSIONS: Our results suggested a cross-sectional independent correlation between the examined cardiovascular risk factors as the predominant features of metabolic syndrome and C-reactive protein in the group of apparently healthy subjects. The lack of correlation of C-reactive protein with the total cholesterol and low density lipoprotein cholesterol in our study may suggest their different role in the process of atherosclerosis and the possibility to determine C-reactive protein in order to identify high-risk subjects not identified with cholesterol screening. PMID- 16375205 TI - [Correlation between findings of echosonography and endoscopic retrograde cholangiopancreatography examination in chronic pancreatitis patients]. AB - BACKGROUND/AIM: Chronic pancreatitis is defined as an amount of chronic inflammatory lesions that lead to the destruction of pancreatic tissue and fibrosis development, whereas the later stages of the illness are characterized by the destruction of the endocrine portion of the organ. Although the results of different studies are abundant, chronic pancreatitis still remains enigmatic, both in its diagnostic and therapeutic aspect. To test the correlation between the findings of echosonography and endoscopic retrograde cholangiopancreatography (ERCP) examination in chronic pancreatitis patients. The observed degree of correlation may serve for the validation of echosonography as a diagnostic tool in chronic pancreatitis patients. METHODS: We collected and analyzed data on morphological features in chronic pancreatitis patients revealed by echosonography as well as endoscopic retrograde cholangiopancreatography. RESULTS: 35 patients 34-73 years of age were included in this study. In 60% (21 subject) history was subjective for alcohol abuse. Significant correaltion has been found between alcohol abuse and chronic pancreatitis (chi2 = 6.896; p < 0.05). Correaltion between groups of chronic pancreatitis patients diagnosed by echosonography and endoscopic retrograde cholangiopancreatography was highly significant (p = 0.799; p < 0.01). CONCLUSION: Echosonography was proved to be a suitable first choice imaging method for the examination of patients when chronic pancreatitis was suspected. Echosonography might provide conclusive information on the morphology of pancreatic canalicular system, as well as on the state of pancreatic parenchyma. PMID- 16375206 TI - [Unerupted lower third molar as a cause of acute infection in soldiers]. AB - BACKGROUND/AIM: To record the frequency of infections as complications accompanying the unerupted lower third molars and to determine if there was a significant level of the relationship between the frequency and the severity of infections and the age of patients. METHODS: This study included 100 soldiers of the Army of Serbia & Montenegro, with the unerupted third molars, of male sex, 18 to 25 years old, who were receiving clinical treatment. RESULTS: In 73% of the patients the infection was caused with the unerupted lower third molars. The highest frequency of infection was observed in the group of 20-23 years of age (75.3%). Mild infection occurred in 49 (67.1%) of the patients. CONCLUSION: The frequency of infection was significantly higher in the older patients. Because of the high frequency of the infection recorded, the unerupted lower third molars should be surgically removed before the age of 20. PMID- 16375207 TI - [Possibility to predict the development of secondary depression in primary alcoholics during abstinence]. AB - BACKGROUND/AIM: [corrected] The relationship between alcoholism and depression is observed in clinical trials. The factors which could predict persistence of secondary depression after alcohol withdrawal are not enough explored on admission. The differencies between depressed (DA) and non-depressed (NDA) alcoholics regarding the degrees of severity of withdrawal, severity of depression and the intensity of cognitive disfunctions were explored on admission to investigate possibility of prediction of the development of secondary depression in alcoholics. METHODS: A group of primary male alcoholics (n = 86) was recruited during inpatient treatment. After 4 weeks alcoholics were divided in the DA group (n = 43) and NDA (n = 43) group according to the score on the Hamilton Depression Rating Scale (HAM-D). Clinical assessment of withdrawal, scoring on the Alcohol Dependency Severity Scale-ASD, and scoring on the Mini Mental Scale-MMSE were performed in all the participants on admission. The differences between the groups were tested by the Student's t-test. RESULTS: The DA group showed the significantly higher severity of depression, higher levels of alcohol withdrawal symptoms and cognitive disfunctions than the NDA group on admission. CONCLUSION: The specific group of depressive alcoholics was shown to be characterized by the higher severity of alcoholism and depression on admission, which could predict prolonged, secondary depression. Early detection and concurrent therapy of secundary depression could improve the treatment, and reduce the relapse of alcoholism. PMID- 16375208 TI - [Step by step to successful oral presentation]. PMID- 16375209 TI - [Population pharmacokinetics]. PMID- 16375210 TI - [Professional burnout]. PMID- 16375211 TI - [Tertiary blast injury to the intestines]. AB - BACKGROUND: To present the first case of the tertiary blast injury to the intestine, and the tertiary blast injury in general. CASE REPORT: A parachutist of the Army of Serbia and Montenegro was injured when descended from the 1 200m height by parachute which did not expand. The force of stroke to the ground, caused the reactive transfer of energy and the subsequent blast injury to the intestine. After 24 hours, the secondary perforation of the small intestine, contusioned by the blast, developed which was the indication of explorative laparotomy. The resectioned small intestine showed the histologic characteristics of a blast injury, so the tertiary blast injury was diagnosed on the basis of these and of the mechanism of the injury. CONCLUSION: Tertiary blast injuries fall into the group of indirect blast injuries. The only difference between indirect injuries as compared to direct ones, is in the manner of inflicting, otherwise the traumatic mechanisms are alike, and include the transfer of the energy of stroke through the tissue of different density. PMID- 16375212 TI - [Neuroendocrine tumor of the skin of head and neck]. AB - BACKGROUND: Merkel cell carcinom is a rare neuroendrocine tumor of skin which manifests it self through aggressive growth and early regional metastasis. It develops mainly in older population. Locally, the tumor spreads intracutaneously. CASE REPORT: We showed two cases (females of 89 and 70 years old) hospitalized within the last two years. The first patient was treated surgically three times. After the surgery, the patient was treated with radio therapy, and died 3 years from the begining of the treatment. The second patient with this neuroendocrine tumor with the high malignacy potential and huge regional metastasis, was treated surgiclly, and died a month and a half after the operation. CONCLUSION: These two cases confirmed the aggressive and recidivant growth of this tumor with the difficut pathologic investigetion, and the extremlly bad prognosis inspite of the treatment. PMID- 16375213 TI - Treatment of anicteric acute hepatitis C with peginterferon alpha-2a plus ribavirin. AB - BACKGROUND: Hepatitis C virus (HCV) infection is the most frequent cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma in the world. Acute hepatitis C is the most commonly asymptomatic liver disease with the development of chronic HCV infection in the majority of infected patients. Studies of the natural history of HCV infection suggest that only 15-30% of patients with acute infection recover spontaneously. Others, up to 85% of the infected patients develop chronic hepatitis C. Acute hepatitis C is so uncommon and with the unpredictable occurrence, and of the low frequency, that it is difficult to determine the optimal treatment of this disease. There have been many randomized, controlled trials of the therapy in patients with chronic hepatitis C, but none of an adequate size or rigor in patients with acute hepatitis C. Therefore, the causal treatment of patients with acute hepatitis C aimed at the prevention of chronic liver disease is necessary. CASE REPORT: We have treated a patient with anicteric form of acute hepatitis C after a three-month outpatient follow-up using a combined therapy: pegylated interferon-alpha 2a, 180 microg, subcutaneously, once a week plus ribavirin 1000 mg orally once a day. The treatment lasted 24 weeks. Stable biochemical and virological response was achieved both at the end of the treatment and 6 months after the completion of the therapy. CONCLUSION: We believe that the above mentioned might be one of the approaches to the treatment of acute hepatitis C. However, further prospective studies with significantly larger number of patients are necessary for the definite conclusions about the treatment of HCV infections. PMID- 16375214 TI - Differential regulation of proopiomelanocortin (POMC) mRNA expression in hypothalamus and anterior pituitary following repeated cyanamide with ethanol administration. AB - BACKGROUND/AIM: We have investigated proopiomelanocortin (POMC) mRNA expression in the arcuate nucleus of the hypothalamus (ARC) and the anterior lobe of the pituitary (AL) following repeated cyanamide-ethanol reaction (CER). METHODS: Adult male Sprague-Dawley rats (250-290 gr) were housed in a temperature and humidity controlled environment with free access to food and water. Four experimental groups were used as follows: saline (as control), cyanamide alone, ethanol alone and ethanol with cyanamide. The animals recived daily intraperitoneal injections (i.p.) of cyanamide (10 mg/kg, 60 min before ethanol dosing) with or without ethanol (1g/kg) for 5 consecutive days, and were sacrificed 60 min after the last dosing of ethanol. The results were presented as the mean +/- SEM for each group. All groups within each deta set were compared by one-way ANOVA followed by Fisher PLSD test for multiple comparisons. A value of p < 0.05 was considered significant. RESULTS: The POMC mRNA levels in ARC were significantly decreased with cyanamide compared to the control and ethanol alone (p < 0.05 and p < 0.05 respectively), but increased in AL following repeated CER. CONCLUSION: We speculate that this differential regulation of POMC mRNA expression may be partially involved in the preventive effects on alcohol intake in response to CER. PMID- 16375215 TI - Expression of p53 protein in Barrett's adenocarcinoma and adenocarcinoma of the gastric cardia and antrum. AB - BACKGROUND/AIM: Most studies of esophageal and gastric adenocarcinomas have shown a very high rate of p53 gene mutation and/or protein overexpression, but the influence of the tumour site upon the frequency of p53 protein expression has not been evaluated (gastroesophageal junction, Barret's esophagus, and antrum). The aim of our study was to analyze the correlation between the selected clinico pathological parameters, and p53 protein overexpression in regards to the particular tumour location. METHODS: The material comprised 66 surgical specimens; 10 were Barrett's carcinomas, 25 adenocarcinomas of the gastric cardia (type II adenocarcinoma of the esophagogastric junction - EGJ), and 31 adenocarcinomas of the antrum. Immunostaining for p53 protein was performed on formalin-fixed, paraffin-embedded tissue sections, using the alkaline phosphatase antialkaline phosphatase (APAAP) method. The cases were considered positive for p53 if at least 5% of the tumour cells expressed this protein by immunostaining. RESULTS: There was no significant difference observed between the studied groups in regards to age, sex, Lauren's classification and tumour differentiation. There was, however, a significant difference observed in the depth of tumour invasion between Barrett's adenocarcinoma and adenocarcinoma of the cardia compared with the adenocarcinoma of the antrum. Namely, at the time of surgery, both Barrett's adenocarcinomas and adenocarcinomas of the cardia, were significantly more advanced comparing with the adenocarcinomas of the antrum. Overexpression of p53 was found in 40% (4/10) of Barrett's adenocarcinomas, 72% (18/25) of adenocarcinoma of the cardia and 65% (20/31) of adenocarcinoma of the antrum. No significant differences in p53 expression in relation to sex, type (Lauren) of tumour, depth of invasion, lymph node involvement, or tumour differentiation were observed in any of the analyzed groups of tumours. Patients with more advanced Barrett's adenocarcinoma and in the cases of lymph node invasion revealed tendency for the greater p53 positivity compared with the early forms and lymph node-negative cases; however, this difference was not significant according to the statistical analysis. With regard to adenocarcinoma of the cardia, higher rates of p53 positivity were recorded in poorly differentiated, more advanced cases with lymph node invasion. Nevertheless, none of these differences was statistically significant. On the contrary, in the patients with adenocarcinoma of the antrum, greater p53 positivity was revealed in early forms without lymph node involvement, but the observed difference was not statistically significant. CONCLUSION: No significant differences in p53 protein expression in terms of sex, type (Lauren) of tumour, depth of invasion, lymph node involvement, or tumour differentiation were observed in any of the analyzed groups of tumours (Barrett's adenocarcinoma, adenocarcinoma of the cardia and adenocarcinoma of the antrum). PMID- 16375216 TI - A pharmacokinetic comparison of three pharmaceutical formulations of nimesulide in healthy volunteers. AB - BACKGROUND/AIM: Switching the patient from one pharmaceutical formulation of the same drug to another, may lead to therapeutic inadequancy in some cases. To minimize the risk, careful pharmacokinetic studies are desired in the pre registration period and afterwards. METHODS: A randomized, crossover design with one-week wash-out period between each dose was applied. Serum samples, obtained before dosing and at various appropriate time points up to 15 hours, were analyzed for nimesulide content by a high-performance liquid chromatographic method with ultraviolet (LU) detection. The pharmacokinetics and relative bioavailability of three different pharmaceutical formulations containing nimesulide, manufactured by the same pharmaceutical factory, were studied prospectively in 12 healthy subjects of both sexes. A single 100-mg oral dose of nimesulide was given to the volunteers in the form of conventional tablets, mouth dissolving tablets or as a suspension. Analysis of variance, power analysis, 90% confidence intervals, and two one-sided tests were used for the statistical analysis of pharmacokinetic parameters. RESULTS: The tolerability of all preparations was excellent. The respective confidence intervals of the ratios of geometric means of C(max) and AUC(0-infinity) of nimesulide were out of acceptable limits either for conventional tablets in comparison with suspension or for mouth dissolving tablets when compared with conventional tablets. A comparison of mouth dissolving tablets with suspension showed a statistically significant difference between C(max) values (suprabioavailability of mouth dissolving tablets), while the point estimate of the ratio of geometric means of AUC(0-infinity) was 0.945 with the corresponding 90% confidence interval of 0.902 0.991. At the 5% level of significance, there were no differences between the formulations under the study in times elapsed to peak serum concentrations, as revealed by the non-parametric Wilcoxon signed ranks test. CONCLUSION: Only a 90% confidence interval for the relative differences of log-transformed AUC(0 infinity) values of nimesulide absorbed from mouth dissolving tablets vs. suspension was included in the 80% to 125% interval proposed by the Food and Drug Administration (FDA). On that basis, mouth dissolving tablets (Nimulid-MD) were considered bioequivalent to Nimulid suspension according to the extent of drug absorption. Concerning the comparable amounts of nimesulide available in the systemic circulation after application of these formulations the one might not expect therapeutic failure after switching the patient from one to another. PMID- 16375217 TI - [Reconstruction of the infected war defects of the tibia: a fragment elongation according to the Ilizarov technique]. AB - INTRODUCTION/AIM: The treatment of tibia defects complicated with chronic osteomyelitis is difficult, often requiring one or more surgical interventions with prolonged periods of functional incapacity. METHODS: We treated 20 patients with tibia defects, who had been wounded during the war operations in the former Yugoslavia, complicated with chronic osteomyelitis by applying the Ilizarov apparatus. In 10 patients with the average defect of 4.7 cm, interfragmentary diastasis of 1.5 cm, and 3.1 cm of shortening we applied the bilocal synchronous compressive - distractive method (BSCD). In the remaining 10 patients with average defect of 6.4 cm, interfragmentary diastasis of 5.5 cm, and 1.6 cm of shortening we applied bilocal alternating distractive - compressive osteosynthesis (BADC). RESULTS: The average followup was 93 months. In the group A, the average distraction index was 10.6, maturation index 39.8, and external fixation index 52.5. In the group B, the average distraction index was 11.7, maturation index 47.1, and external fixation index 60.1. The average time from the application to the apparatus removing was in the group A 6.5 months, and in the group B 11.9 months. There were 27 complications: 11 problems, 8 disturbs, and 8 true complications. Pin-track inflammation of the soft tissue was noted most frequently (6 patients). CONCLUSIONS: One stage of repairing inflamation and the restitution of defect in lower leg tissue was the advantage of this type of treatment. All of the patients recovered. There was not any bad result, either in osteal or in functional outcome. PMID- 16375218 TI - [Significance of ultrasound and ultrasound-guided fine-needle aspiration for the detection of laryngeal occult metastases]. AB - BACKGROUND/AIM: To evaluate ultrasound criteria based on a node size, shape, vascularity and cytology findings with respect to their value for the comparative determination of metastatic lymph nodes in laryngeal carcinoma. METHODS: A prospective study included 30 patients with laryngeal squamous cell carcinoma without node enlargement on computerized tomography, at the Department of Otorhinolaryngology, the University Hospital, Zemun. Thirty-six neck lymph nodes were evaluated sonographically and aspirated with an ultrasound-guided fine needle. They were examined cytologically and/or histopathologically and compared to the sonographic assessment of their malignancy. RESULTS: Of the 36 neck lymph nodes evaluated cytologically, the 13 were found to be with a metastatic deposit. The assessment of a lymph node malignancy using the parameter of size had the senzitivity of 84%, the specificity of 70%, and the reliability of 75%. Using the criteria of a lymph node shape for the assessment of malignancy, the sensitivity of 61%, specificity of 65%, and the reliability of 64% were achieved. The evaluation of a lymph node vascularity by the use of the effect of Doppler showed the sensitivity of 69%, the specificity of 95%, and the reliability of 86%. CONCLUSION: Ultrasound and ultrasound-guided fine-needle aspiration citology should be always used for the preoperative staging and for the postoperative follow-up of the status of the neck with cancer of the larynx becouse of their high accuracy, availability and semiinvasivity, and in order to enhance the reliability of the evaluation of the malignant disease progression. PMID- 16375219 TI - [The aqueous humour antioxidative capacity in different types and color of the age-related cataract]. AB - BACKGROUND/AIM: Oxidative stress results from increased oxidative processes, decreased antioxidative protection, or both processes simultaneously. Photooxidative stress, as a form of oxidative stress, induced by the energy of solar radiation, today is considered as crucial in the age-related cataractogenesis. Other known and unknown, endogenous and egsogenous factors that contribute to the oxidative stress intensity, can influnce the cataract type and brunescence. Thus the oxidative stress intensity and its form might determine the cataract type and brunescence, and also make the efforts in cataract prevention more complex. Hence, the objective of the present paper was to investigate the current amount of antioxidative capacity in aqueous humour during the cataract genesis of different types and pigmentation of cataract. METHODS: Transversal review of 80 samples of humour aqueous obtained during extracapsular cataract extraction. Aqueouses were analyzed by tiobarbituric acid (TBA) method for the total antioxidant activity estimation, expressed as %iMDA, and by using 0.1 ml of aqueous. RESULTS: The mixed type of cataract showed the statistically significantly lower values of the intensities of antioxidative protection in aqueous humour compared to cortical and nuclear cataracts (p < 0.001, respectively). Between pure nuclear and cortical cataracts we found the small differences of the investigated parameter, but they pointed to the decreased level of antioxidative protection, i.e. the increased intensity of the aqueous humour oxidative stress in the cortical cataract type. A significant correlation betweer the cortical cataract maturation and the %iMDA (p < 0.05) was found. CONCLUSIONS: The role of the oxidative stress, here expressed as the antioxidative capacity of aqueous humour, could not be the same for all the cataract types. The lower level of antioxidative protection of aqueous in brunescent and mixed cataracts may point to the higher intensity of oxidative stress in those cataract types. The correlation betweer the cortical cataract maturity and %iMDA points to the significant decrease of the aqueous antioxidative protection in the cataract progression. PMID- 16375220 TI - [Frequency of metastatic tumors in the heart]. AB - INTRODUCTION: Secondary or metastatic tumors in the heart occur more frequently than primary ones, and, according to the published series, their frequency found in autopsic material ranges from 1.6% to 20.6%. Metastatic tumors in the heart are rarely clinically symptomatic, and, therefore, they are rarely diagnosed within the lifetime. They are mostly diagnosed at autopsy. The aim of this study was to analyze the frequency of metastatic tumors of the heart, their primary localization, as well as the localization of the metastases found in the autopsic material within the period 1972-2004. METHODS: During the autopsy of the patients died of metastatic tumors, we microscopically and macroscopically analyzed all the organs and tissues to determine the metastases of primary tumors in other organs, especially in the heart and pericardium. RESULTS: Within the period from 1972-2004, 11 403 autopsies were performed. In 2 928 (25.6%) out of 11 403 autopsies, the presence of malignant tumor was diagnosed, and in 79 (2.7%) of these cases, metastasis of the heart was found out. Only in 5 of the cases, the presence of metastasis in the heart was diagnosed during the lifetime. The most frequent metastases in the heart were caused by pulmonary carcinoma (18 cases), leukemia and malignant lymphoma (8 cases, each), then pancreatic and breast carcinoma, while the metastases of other carcinomas were rather rare. In 40 (60.76%) cases, the metastasis was localized in the myocardium, but more often in the left ventricle, in 24 (30.38%) cases in the pericardium, in 4 cases in the epicardium and in the 3 of them in the mitral and tricuspid valve. Only in one case of renal carcionoma, metastasis was found in the right atrium and it occurred by spreading (dissemination) through the lumen of the inferior vena cava. CONCLUSION: Metastatic tumors of the heart are rather rare, and rarely clinically symptomatic, and, thus, rarely diagnosed during life. The methods of choice for the diagnosis of the metastasis in the heart are echocardiography, computerized tomography, magnetic resonance imaging, cytological analysis of the pericardial effusion and biopsy. The treatment includes surgery, chemotherapy and radiotherapy. PMID- 16375221 TI - [Influence of proteinuria on the lipoprotein (a) metabolism disorder]. AB - BACKGROUND/AIM: Proteinuria causes lipid metabolism abnormalities. The aim of the present study was to examine the influence of proteinuria on the lipoprotein (a) metabolism disorder. METHODS: The study included 60 patients of the male-famele ratio (M: F = 32 : 28), mean age 37.15 +/- 9.85 years with, the average endogenous creatinine clearance 86.27 +/- 19.81 ml/min, and the average body mass index (BMI) 24.18 +/- 2.23 kg/m2. Regarding the level of glomerular proteinuria, the patients were divided into four groups. The first (control) group, with proteinuria levels less than 0.25 g/24h, included 15 patients (M: F = 6 : 9), mean age 34.66 +/- 4.82 years, the mean clearance of endogenous creatinine 99.70 +/- 12.94 ml/min, and mean BMI 23.28 +/- 3.50 kg/m2. The second group, with proteinuria between 0.25 and 1.0 g/24 h, includ 15 patients (M: F = 9 : 6) with primary glomerulonephritis, mean age 37.87 +/- 9.65 years, the mean clearance of endogenous creatinine 82.85 +/- 18.48 ml/min, and mean BMI 23.83 +/- 1.57 kg/m2. The third group includ 15 patients (M : F = 8 : 7) with primary glomerulonephritis, with proteinuria between 1.0 and 3.0 g/24 h, mean age 35.67 +/- 13.29 years, the mean clearance of endogenous creatinine 82.85 +/- 18.48 ml/min, and mean BMI 23.83 +/- 1.57 kg/m2. The fourth group, with proteinuria higher than 3.0 g/24 h, included 15 patients (M: F = 9 : 6) with primary glomerulonephritis, mean age 40.40 +/- 9.75 years, the mean clearance of endogenous creatinine 80.16 +/- 20.80 ml/min, and mean BMI 24.83 +/- 1.44 kg/m2. In order to assess the influence of proteinuria on the lipoprotein (a) metabolism abnormalities we investigated 24-hour proteinuria, the colloid osmotic pressure (COP) of plasma, and the serum concentration of lipoprotein (a). The results were statistically analyzed using ANOVA, Kruscal-Wallis test, Mann-Whitney U test, chi2 test and Spearman test. RESULTS: Statistically, the patients with proteinuria over 3.0 g/24 h had the significantly higher values of lipoprotein (a) in serum as compared to the control group, and the patients with proteinuria about 0.25-1.0 g/24 h. The patients with proteinuria between 1.0-3.0 g/24 h had the statistically significantly higher values of lipoprotein (a) in serum than the control group (proteinuria < 0.25 g/24 h). There was a highly statistically significant negative correlation between serum albumin concentration, COP and the concentration of lipoprotein (a) in serum. There was a highly statistically significant positive correlation between 24-hour proteinuria and the concentration of lipoprotein (a) in serum. CONCLUSION: Proteinuria leads to the deterioration of lipoprotein (a) abnormalities. PMID- 16375222 TI - [Prolonged survival of a female patient with total pelvic exenteration]. AB - BACKGROUND: To present a female patient who lived 5 years after total pelvic exenteration (TPE). CASE REPORT: The female patient underwent TPE due to retrovesicovaginal fistula as a consequence of locoregional irradiation after the operation for the malignoma of the vaginal part of the uterus. In the formation of Bricker conduit, the ureter antireflux was achieved by the application of the "tobacco sack muff" made of the intestines around the ureter. By the use of this technique, the occurrence of pyelonephritis, as the leading cause of death in such patients, was prevented. CONCLUSION: TPE is a hope for significantly prolonged survival of patients with advanced pelvic malignomas, or with a postirradiatiation fistula. PMID- 16375223 TI - [Uretero-transappendix-cystoneostomy as a technique for the reconstruction of the lower part of the ureter]. AB - BACKGROUND: Any large missing part of the ureter may be replaced by transureteroureterostomy, psoas hitch, Boari flap, nephrectomy, renal autotransplanation or by the implementation of an intestinal graft. CASE REPORT: A patient with a defect of the lumbal-pelvic portion of the right ureter, after the managemenet of a penetrating and perforating gun shot wound was presented. The missing part of the ureter was seccessfully replaced with an appendix. The technique of uretero-transappendix-cystoneostomy complete with a Boari flap and a psoas hitch was used. CONCLUSION: By the use of an original combination of surgical techniques, a large defect of the ureter and the defect of the bladder, as well as the preservation of the renal function was achieved in a more successful manner. PMID- 16375224 TI - Metabolic syndrome and central retinal artery occlusion. AB - BACKGROUND: The accumulation of risk factors for central retinal artery occlusion can be seen in a single person and might be explained by the metabolic syndrome. CASE REPORT: We presented the case of a 52-year-old man with no light perception in his right eye. The visual loss was monocular and painless, fundoscopy showed central retinal artery occlusion and the laboratory investigation showed the raised erythrocyte sedimentation rate of 105 mm/h and the raised C-reactive protein of 22 mg/l. Specific laboratory investigations and fluorescein angiography excluded the presence of vasculitis, collagen vascular diseases, hypercoagulable state and antiphospholipid syndrome. CONCLUSION: The patient met all the five of the National Cholesterol Education Program (NCEP) criteria for the metabolic syndrome: hypertension, abnormal lipid profile, abnormal glucose metabolism, obesity and hyperuricemia. Measurement of C-reactive protein is useful for the assessment of therapeutic systemic effect on any abnormality in the metabolic syndrome. Individual therapy for all risk factors in the metabolic syndrome is necessary to prevent complications such as cardiovascular, retinal vascular diseases and stroke. PMID- 16375225 TI - [Half a century of electron microscopy in Serbia]. PMID- 16375226 TI - [Ignaz Philipp Semmelweis (1818-1865)]. PMID- 16375227 TI - Psoriatic arthritis. PMID- 16375228 TI - FEM in periodontal research. AB - The changes in the periodontal tissues either due to pathologic processes on external forces have been investigated from histologic, histochemical, biochemical, physiologic, and bioelectrical (bony response) and biochemical viewpoint. However, because of the complexity of stress analysis or experimental techniques, no information is available on the stress induced in the periodontal tissues. The Finite Element Method (FEM), which is introduced as one of the methods for numerical analysis, has become a useful technique for stress analysis in biologic systems. The FEM makes it possible to analytically apply various force systems at any point and in any direction. From the periodontal point of view, FEM would provide different stress levels in periodontal structures based on the biomechanical principle, which in turn may prompt occlusal correction at an earlier date. PMID- 16375229 TI - Immunohistochemical evaluation of expression of cytokeratin 19 in different histological grades of leukoplakia and oral squamous cell carcinoma. AB - Recent progress in understanding the biology of keratins together with the development of monoclonal antibodies to individual keratin proteins provide the foundation for studying the keratin expression in normal and pathological oral epithelia. Cytokeratin (CK) alterations have been reported in carcinomas and these have been associated with specific aspects of tumour behaviour. Immunohisto chemistry with monospecific CK19 antibody was used to study the expression pattern in normal mucosa, dysplasias, and oral squamous cell carcinomas (OSCC). In non-keratinzed normal mucosa, CK19 was detected in the basal cell layer, while in dysplasias (diagnosed in H and E stained sections, mild-severe) stained strongly for CK 19 in the basal and supra basal cell layers indicating layer specificity for CK 19 expression. In OSCC, in the number of CK19 labelled cells increased from well to poorly differentiated tumour. Thus the results of the present study indicate an alteration in synthesis of keratin proteins when exposed to carcinogens. PMID- 16375230 TI - Histopathological study of apparently normal oral mucosa in lepromatous leprosy. AB - In lepromatous leprosy, it may be postulated that apparently normal areas of skin are the probable sites of future extension of the pathological process and the events taking place in these site might represent the earlier stages of such an extension. Thus the present study has been done to evaluate any such changes in the apparently normal oral mucous membrane of the lepromatous leprosy patients, since oral involvement is common in these patients. Twenty two cases of lepromatous leprosy were studied histologically to see changes in apparently normal oral mucosa. Out of the 22 patients, apparently normal skin biopsies were taken from 13 patients and 5 lesional biopsies were taken from the same patients. Apparently normal oral mucosal biopsies were done for all patients. Biopsies of 22 cases of apparently normal oral mucosa did not show granulation formation of acid-fast bacilli histologically. The only change present was mild inflammation. PMID- 16375231 TI - Retromolar foramen. AB - Of the many variants in the mandible, the retromolar foramen is the most important. The incidence of retromolar foramen was studied in a collection of 157 south Indian mandibles available at the department of Anatomy, St. John's Medical College, Bangalore. On each side of the mandible, the area behind the last molar tooth was inspected for the presence of retro molar foramen. Wherever present, their distance from the last molar tooth and their size if they were sufficiently large were measured with a sliding caliper to the nearest millimeter. Retromolar foramen were found to be bilateral in 8 mandibles (5.1%) and unilateral 20 [right: 9 (5.7%); left: 11 (7%)] mandibles. The diameter of the foramina was 2 mm on both sides. PMID- 16375232 TI - An association of external and internal enamel pearls. AB - We report a rare case of an association of external enamel pearl with internal enamel pearl on the root of a molar. To the best of our knowledge, association of external and internal enamel pearls has not been previously reported. We discussed the histogenesis of enamel pearls and proposed that internal enamel pearl formation may be a continuation of formation of external enamel pearl. PMID- 16375233 TI - Ameloblastic fibro odontoma. AB - Ameloblastic fibro odontoma is relatively a rare mixed odontogenic tumor comprising of ameloblastic fibroma and ameloblastic odontoma. This group of lesions represent neoplastic and hamartomatous changes. Data regarding age, sex, and site of occurrence of these lesions have been uniquely described. A case of ameloblastic fibro odontoma in a 17-year-old male involving the left body of the mandible, and its radiographic, clinical symptoms and histological features is described herewith. A conservative surgical approach, including enucleation and mechanical curettage of the surrounding bone was done. PMID- 16375234 TI - Juvenile nasopharyngeal angiofibroma. AB - Juvenile nasopharyngeal angiofibroma is a rare, histologically benign yet locally aggressive, vascular tumor that typically affects adolescent males. It accounts for 0.5 percent of all neoplasms of the head and neck. A case of juvenile nasopharyngeal angiofibroma manifesting in the oral cavity in a 20-year-old male patient is presented and discussed. PMID- 16375235 TI - Lobster-claw syndrome. AB - Lobster-Claw syndrome is a rare autosomal dominant, hand-foot malformation with Oro-dental features. This is a rare condition and from the available reports so far, cases being reported are less than 1%. Most reports have focused on the hand foot deformity of this syndrome. This paper highlights the typical Oro-dental features associated with this syndrome such as retained deciduous teeth, hypodontia and variation in crown size, arch length and arch width. PMID- 16375239 TI - [How to take charge of osteoporosis in women in 2005]. PMID- 16375240 TI - [Controlled analgesia pumps]. PMID- 16375241 TI - [Type 2 diabetes: why screen, how to prevent?]. PMID- 16375242 TI - [Insulin pumps]. PMID- 16375243 TI - [Oral hygiene]. PMID- 16375245 TI - [Nursing expert before the tribunal. Interview by Delmotte Helene]. PMID- 16375244 TI - [Students' stress when facing death]. PMID- 16375246 TI - [Public health reform. Law no. 2004-806 of August 9, 2004 relative to public health policy]. PMID- 16375247 TI - [Weight gain and stress]. PMID- 16375248 TI - State of the art analytical comparability: a review. PMID- 16375249 TI - Validation of peptide mapping with electrospray mass spectrometry for recombinant proteins of biopharmaceutical interest and its applications as an identity test and a characterization tool. AB - In this paper, the steps required to validate a liquid chromatography peptide mapping method with mass spectrometric detection (LC-MS) for use as an identity test and characterization tool are presented. All aspects of peptide mapping are evaluated and optimized, including protein sample preparation (protein reduction, alkylation and enzymatic digestion), high performance liquid chromatography (HPLC) separation of the resulting peptides, and the use of a mass spectrometric detection. In addition, the validation of a single quadruple MS detector is described and the implementation of on-line electrospray ionization MS (ESI-MS) as an adjunct detector to support the investigation of peak differences is presented. Applications of peptide mapping with tandem MS using an electrospray ion-trap instrument throughout the biopharmaceutical product development cycle are discussed, including assessing protein product heterogeneity derived from post-translational modifications (e.g. multiple N- or C-termini, deamidation, oxidation and glycosylation) and protein degradation. PMID- 16375250 TI - Analysis of protein therapeutics by capillary electrophoresis: applications and challenges. AB - Capillary electrophoresis (CE) has been increasingly used for the analysis of recombinant protein therapeutics in the biotechnology industry over the past several years. In this paper, an overview of the major applications implemented at Genentech Inc. is presented. The applications highlighted in this article are divided into the following three general areas: (i) CE as a replacement for slab gel electrophoresis, particularly capillary electrophoresis-sodium dodecylsulphate and capillary isoelectric focusing; (ii) CE to monitor protein charge heterogeneity as an orthogonal technique to the traditional on-exchange chromatographic methods; and (iii) CE for carbohydrate analysis, including both oligosaccharide and monosaccharide analysis. Overall, the advantages of these CE based methodologies include automation, ease of quantification, rapid analysis time, enhanced resolution, and robustness when compared to the traditional methods. There are, however, still some challenges in applying CE for protein analysis, particularly in the area of characterization due to the miniaturization nature of CE. PMID- 16375251 TI - Mass spectrometry and HPLC for carbohydrate analysis. AB - Methods have been developed for determining glycan galactosylation of recombinant monoclonal antibodies based on a number of techniques including: electrospray ionization mass spectrometry, matrix-assisted laser-desorption time-of-flight mass spectrometry, as well as fluorescent labelling and analytical separation of released glycans. While the techniques used differ in the analytical approaches, they all yield highly reproducible, linear and precise data for IgG glycoform analysis. PMID- 16375252 TI - Using nuclear magnetic resonance spectroscopy to characterize biologicals. PMID- 16375253 TI - Infrared and Raman spectroscopy characterization of biological products. AB - Both infrared and Raman are increasingly applied in biomolecular structure characterization, thus gaining acceptance alongside traditional bioanalytical techniques. Physical principles and biological sampling requirements are reviewed. Advanced applications are used to illustrate vibrational spectroscopy capabilities and limitations in bioanalysis. PMID- 16375254 TI - Characterization of an immune response. AB - The characterization of any detected antibodies provides detailed information that can be critical to understanding the significance of an immune response. The BIAcore immunoassay provides a straightforward means of characterizing many aspects of the immune response including antibody concentration, specificity, binding affinity, and the presence of isotypes. One important aspect of the immune response is determination of neutralizing capability, which requires a biological assay. The ultimate clinical significance of an immune response can only be fully understood when data from antibody characterization are coupled with clinical data from the patient. Some of the factors that can contribute to the clinical significance of an immune response are: (i) magnitude of the immune response (concentration of antibodies detected); (ii) duration of the immune response (continuous antibody production or sporadic and not sustained); (iii) correlation with any adverse events; (iv) correlation with a change in pharmacokinetics (either mediating sustained circulation or enhanced clearance of the drug); (v) biological neutralization of the drug; (vi) biological neutralization of an endogenous protein. Full characterization of the immune response requires the incorporation of antibody assays, pharmacokinetic assays, and clinical data. PMID- 16375255 TI - Evaluation of standard and state of the art analytical technology-bioassays. AB - The development of biological assays for assessing potency is a critical component for monitoring the quality of therapeutic biologicals. Traditional cell based bioassays, which are the most widely used, are typically based on a terminal cellular response such as cell proliferation or inhibition. While these assays can be very user-friendly, results often take days and sensitivity is sometimes not sufficient for the needs of the development programme. Recent improvements in analytical technology have led to new approaches in bioassay development. Many of these assays exploit cell signalling pathways far upstream from a terminal cellular response. Bioassays based on a cell signal are much more rapid, sensitive, and indicate stability better than their predecessors. Many of these newer assays are "hybrid" assays which combine the receptor signalling of traditional bioassays with the sensitivity of detection found in immunoassays. One such method, the Kinase Receptor Activation Assay (KIRA), works through the detection of receptor phosphorylation following analyte stimulation. Validations of newer technology assays, such as KIRA, require an individualized strategy due to their unique attributes. A thorough assessment of robustness should be paramount in the validation of these assays. Several examples of new technology platforms for bioassays are also discussed. PMID- 16375256 TI - Heterogeneity of recombinant antibodies: linking structure to function. AB - Structural heterogeneity of recombinant IgG1 antibodies derives from variations in conserved as well as unique structural features. Common sources of heterogeneity include Fc glycosylation, partial heavy chain C-terminal Lys processing, Fc methionine oxidation, hinge-region cleavage, and the glycation of Lys residues. Aspartate residues that are isomerized to iso-aspartate were detected by cation exchange or hydrophobic interaction chromatography for trastuzumab and omalizumab, respectively. Unpaired cysteines were detected in omalizumab using Ellman's reagent, with the thiol-containing Fab resolved using hydrophobic interaction chromatography after papain digestion. Structural variations that cause chromatographic resolution may indicate the presence of a form with reduced potency. PMID- 16375257 TI - Quality control system to support comparability assessment for the Renal Bio Replacement therapy system. AB - Cell therapies offer great promise for treating serious and life-threatening diseases, as cells provide the dynamic, interactive therapy that responds to the patient's condition. The spectrum of cell therapies is very broad, but each requires characterization during product development. During the development of a cell therapy, changes are made to improve product quality, purity, yield, and consistency, as well as to meet other production requirements. Multiple factors have to be considered when changes are made. A well thought-out design of comparability protocols has to reflect the international consensus set forth by the International Conference on Harmonization (ICH) and Food and Drug Administration (FDA). Comparability studies, due to their importance for the entire product development, constitute an additional challenge for the Quality System, which supports manufacturing. A comparability study was conducted to support the transfer of a cell-based technology, the Renal Bio-Replacement (RBR) system, from an academic laboratory to a contract manufacturing facility. The quality control programme supporting the RBR, and issues encountered in the comparability of the RBR therapy system, are discussed in this article. PMID- 16375258 TI - Characterization of gene therapy products and the impact of manufacturing changes on product comparability. AB - Gene therapy products represent a novel and complex class of products. Ensuring product safety, identity, purity and potency following a manufacturing change extends not only to assessing the final formulated product but also all the components used during product manufacturing. CBER has implemented a stepwise approach to product characterization and compliance with cGMPs, which increases as the study moves from phase 1 toward phase 3 and licensing. It is important that product characterization be performed early in product development because without full product characterization it will be difficult to determine the impact of the manufacturing process on the product as well as the impact any manufacturing change will have on the product. To demonstrate product comparability a thorough understanding of the manufacturing process, including product characterization, is necessary, so that the impact of a manufacturing change can be accurately assessed. PMID- 16375259 TI - Characterization of allergen extracts. AB - Allergen vaccines are complex extracts of natural products, and are used for the diagnosis and treatment of allergic diseases. In the U.S., 19 allergen extracts have been standardized. For these vaccines, the potency is estimated by the skin test responses of highly allergic individuals, and surrogate in vitro tests are established for lot release and quality control. The surrogate tests differ for different extracts. National reference standards to which manufactured lots are compared are maintained at FDA/CBER. Allergen standardization has facilitated the establishment of data-driven release limits. PMID- 16375260 TI - Detection, measurement and characterization of unwanted antibodies induced by therapeutic biologicals. AB - Assessment of the unwanted immunogenicity of therapeutic biologicals in recipients is an important consideration in the evaluation of these medicinal products. Proper planning of immunogenicity studies with appropriately devised strategies is critical if valid and meaningful conclusions concerning the unwanted immunogenicity are to be derived. An essential requisite for such studies is the need for conducting carefully selected and validated procedures. Several techniques are available for detection, characterization and measurement of antibodies elicited in an immune response. These include various formats of immunoassays, surface plasmon resonance and biological assays. None of these assays alone can provide sufficient information on the characteristics of the induced antibodies. A combination of methods is therefore usually necessary for a detailed understanding of the quantity and type(s) of antibodies generated against a therapeutic product. This manuscript considers the benefits and limitations of the various techniques available for antibody detection and outlines a brief strategy for the assessment of unwanted immunogenicity of therapeutic products. PMID- 16375261 TI - De-immunization of therapeutic proteins by T-cell epitope modification. AB - Many therapeutic proteins in clinical use have been shown to elicit antibody responses which in some cases have been linked to adverse events. Conventional animal models, although convenient, have rarely been predictive of immunogenicity in humans. New methods for predicting the potential immunogenicity of therapeutic proteins are needed. This treatise proposes a new approach which pairs in silico T-cell epitope analysis with in vitro studies. T-cell epitope mapping algorithms such as EpiMatrix can be used to evaluate a candidate therapeutic protein for T helper epitopes, followed by confirmation of the T-helper epitopes using in vitro methods such as MHC binding assays and T-cell assays. Once these are identified, substitution of key amino acids in the T-cell epitopes may attenuate the immunogenicity of the protein, since modification of the amino acids in anchor position(s) can abrogate binding to human class II MHC molecules and presentation of the peptides, in the context of MHC, to T-helper cells. Following substitution of the key amino acids, immunogenicity of the modified protein can be evaluated in vitro. In parallel, the potential effect of the modifications on the structure of the protein can be evaluated using in silico modeling methods. This multi-step process has been termed DeFT for de-immunization of functional therapeutics. In this article we review the rationale for the approach, provide several retrospective examples that prove the approach in principle, and describe potential applications to therapeutic protein design. The demand for pre-clinical means of evaluating therapeutic proteins is expected to increase with the number of therapeutic proteins and monoclonal antibodies entering the pre-clinical pipeline. Examples provided offer some preliminary proof that the de-immunization approach may improve clinical outcomes. PMID- 16375262 TI - Strategies for the preclinical and clinical characterization of immunogenicity. PMID- 16375263 TI - How are oncology nurses meeting the genetic education and counseling needs of patients? PMID- 16375264 TI - Researchers delve deeper into the human genome. PMID- 16375266 TI - ONS joins forces with the Susan G. Komen foundation to bolster the nation's nursing workforce. PMID- 16375265 TI - Genetic discrimination: it could happen to you. PMID- 16375267 TI - Novel structural motifs in clusters of dipolar spheres: knots, links, and coils. AB - We present the structures of putative global potential energy minima for clusters bound by the Stockmayer (Lennard-Jones plus point dipole) potential. A rich variety of structures is revealed as the cluster size and dipole strength are varied. Most remarkable are groups of closed-loop structures with the topology of knots and links. Despite the large number of possibilities, energetically optimal structures exhibit only a few such topologies. PMID- 16375268 TI - Removing critical errors for DFT applications to transition-metal nanoclusters: correct ground-state structures of Ru clusters. AB - As ruthenium plays an important role in heterogeneous catalysis, understanding the structural and electronic properties of Ru clusters is crucial to advancement of technology. Because of its efficiency, density functional theory (DFT) calculations are often utilized in nanoscience, but careful validation is necessary. Recently, small, nonmetallic Ru(n) clusters were reported by Zhang et al. [J. Phys. Chem. B 2004, 108, 2140] to form unusual square and cubic ground state structures within DFT by treating the exchange-correlation (XC) functional at the level of general-gradient-corrected approximation (GGA). For such clusters, we show that the calculated, energetically preferred structures are sensitive to which XC functional is used and whether relativistic effects are included. We find that a hybrid XC functional with partially exact exchange, such as PBE0, corrects the Ru2 magnetic moment, bond length, and dissociation energy in agreement with experiment and high-level quantum chemistry calculations and changes the Ru4 ground-state structure to a tetrahedron, instead of a square. The change in structural preference is explained by the corrections to the electronic structure of a Ru atom, where the relative position of majority spin s level is shifted with respect to e(g) levels. We also find that standard nonrelativistic DFT-GGA gives similar results to relativistic DFT-PBE0, i.e., relative shifting of s level, but not for the right reasons. Our results again stress the need to validate an XC functional before application to transition-metal nanoclusters. PMID- 16375269 TI - Transport rates vary with deposition time in dip-pen nanolithography. AB - By patterning with dip-pen nanolithography using tip dwell times ranging from 15 s to 2 h over a period of 19 h, we show that the transport rate for smaller patterns is different than for larger ones. This transport behavior is found for both 1-octadecanethiol (ODT) and 16-mercaptohexadecanoic acid (MHDA) inks on gold substrates. Additionally, MHDA shows an overall decrease in transport rate as a function of total writing time during such experiments. These results indicate that measurements with short dwell times are insufficient to determine transport rates for larger features. PMID- 16375270 TI - Electrical behavior of molecular junctions incorporating alpha-helical peptide. AB - We synthesized an alpha-helical peptide containing two terminal thiol groups and demonstrated the method of preparation of a self-assembled monolayer (SAM) on gold with uniform orientation of the molecules on the surface. The monolayers were employed as model systems for the investigations of mediated electron transfer. The measurements of electron transfer efficiency through the peptide were performed using scanning tunneling spectroscopy (STS). The molecules were trapped between the gold tip and the substrate using a Au-S linkage. The electron transfer behavior of the peptide was examined as a function of the tip-substrate distance at fixed bias voltage and as a function of bias voltage at a fixed distance between the tip and the substrate. The data obtained from these experiments indicated that the electron transfer through alpha-helical peptide is very efficient, and its conductivity is comparable to those observed for dodecanedithiol. There is also a directional dependence of electron transmission through the peptide, which is connected with the electric field generated by the molecular dipole of the helix. PMID- 16375271 TI - Synthesis of gold nanoparticles by laser ablation in toluene: quenching and recovery of the surface plasmon absorption. AB - Gold nanoparticles are synthesized by laser ablation of a gold plate in toluene. The nanoparticles do not show their characteristic surface plasmon absorption (SPA) and are found to be included in a graphitic matrix. The absence of this absorption is found to derive from the presence of the matrix which prevents the growth of large nanoparticles and covers them, suppressing the SPA according to the Mie model for core@shell particles. It is possible to recover the nanoparticle SPA by oxidizing the carbon matrix, obtaining, therefore, some control on the activity of this absorption. PMID- 16375272 TI - Fabrication of interconnected 1D molecular lines along and across the dimer rows on the Si(100)-(2 x 1)-H surface through the radical chain reaction. AB - To realize nanoscale wiring in two dimensions (2D), the fabrication of interconnected one-dimensional (1D) molecular lines through the radical chain reaction of alkene molecules (CH2=CH-R) on the H-terminated Si(100)-(2 x 1) surface have been investigated using scanning tunneling microscopy (STM) at 300 K. By the judicious choice of R in the CH2=CH-R molecule and by creating a dangling bond (DB) at a desired point using the STM tip, the perpendicularly connected allyl mercaptan (ALM) and styrene lines have been fabricated on the Si(100)-(2 x 1)-H surface. The continuous growth of the styrene line at the end DB of a growing ALM line (or vice versa) does not occur, perhaps, due to steric hindrance or/and interaction between adsorbed molecules. PMID- 16375273 TI - Self-folding of three-dimensional hydrogel microstructures. AB - This letter describes the fabrication of three-dimensional particulate-like hydrogel microstructures using a combination of soft lithography and volume expansion induced self-folding. Bilayer structures are produced by solvent casting and photocuring of liquid resins. They curl into three-dimensional (3D) structures upon contacting with water due to differential swelling of the two layers. The curvature can be controlled by adjusting the polymer composition of the primary swelling layer. A simple semiempirical mathematical model is used to predict this self-folding behavior. By designing the two-dimensional (2D) shapes of the bilayers, this technique can lead to complicated 3D microstructures. PMID- 16375274 TI - Single-molecule fluorescence imaging of the remote TiO2 photocatalytic oxidation. AB - The remote TiO2 photocatalytic oxidation reaction of single dyes has been investigated by the single-molecule fluorescent imaging technique. The present results suggest that the active oxygen species (Ox) is most probably the .OH radical, which is generated from the photodecomposition of H2O2 by UV light. The analyses of the number, intensity, and spectrum of individual fluorescence spots at the single-molecule level also indicate that unoxidized and oxidized dyes exist during the bleaching processes of single dyes. PMID- 16375275 TI - Ionic limiting molar conductivity calculation of Li-ion battery electrolyte based on mode coupling theory. AB - A method is proposed based on mode coupling theory in which the ion transference number is introduced into the theory. The ionic limiting molar conductivities of LiPF6, LiClO4, LiBF4, LiCF3SO3, Li(CF3SO3)2N, LiC4F9SO3, and LiAsF6 in PC(propylene carbonate), GBL(gamma-butyrolactone), PC(propylene carbonate)/EMC(ethylmethyl carbonate), and PC(propylene carbonate)/DME(dimethoxyethane) are calculated based on this method, which does not involve any adjustable parameter. The results fit well to the literature data which are calculated by an empirically adjusted formula. This presents a potential way to calculate the conductivities of Li-ion battery electrolytes. PMID- 16375276 TI - Single molecule study of perylene orange photobleaching in thin sol-gel films. AB - The paper reports on photobleaching mechanisms of perylene orange embedded in thin sol-gel films, derived from single molecule studies. The experimental configuration uses wide-field illumination and one photon excitation of the molecules. Measurements have been performed both at ambient conditions and under vacuum in order to get information on the influence of oxygen on photobleaching in such porous samples. We have also recorded the evolution of photobleaching with respect to the excitation intensity. The results demonstrate that photobleaching from excited states higher than the first singlet and triplet states has a nonnegligible contribution as soon as the excitation energy exceeds a few hundred W/cm2 and that this process is favored in the presence of air. The study also demonstrates that perylene orange in sol-gel films is not a very efficient emitter but that photobleaching can be slow, which explains the interest for perylene orange as a good candidate to produce long lifetime solid state lasers when embedded in monoliths of sol-gel. PMID- 16375277 TI - 23Na, 29Si, and 13C MAS NMR investigation of glass-forming reactions between Na2CO3 and SiO2. AB - The glass-forming reactions between sodium carbonate (Na2CO3) and silica (SiO2) have been investigated by 23Na, 29Si, and 13C magic-angle spinning (MAS) NMR spectroscopy. The multinuclear MAS NMR approach identifies and quantifies reaction products and intermediates, both glassy and crystalline. A series of powdered batches of initial composition Na2CO3.xSiO2 (x = 1, 2) corresponding to a sodium metasilicate (Na2SiO3) and sodium disilicate (Na2Si2O5) stoichiometry were investigated after periods of isothermal and nonisothermal heat treatments at different temperatures. Analysis of the 23Na quadrupolar coupling parameters has identified the early reaction product in all cases as crystalline Na2SiO3. In the nonisothermal experiment, this reaction is preceded by an early silica-rich melt phase formed around 850 degrees C. The early reactions are controlled by solid-state Na+ diffusion across the reaction zone in the grain interface layer. Crystalline Na2SiO3 precipitates in the interface layer, increasing its thickness between the Na2CO3 and the SiO2 grains and slowing down the rate of Na+ migration. This creates a secondary phase, which is temperature dependent. At low temperatures, where Na+ migration is impaired, the production of Na2SiO3 ceases and silica-richer phases are precipitated. In the case of the sodium disilicate batch, where excess SiO2 is present, a secondary reaction of Na2SiO3 with SiO2 forming a glassy phase is observed. A transient carbon-bearing phase has been identified by 13C NMR as a NaCO3- complex loosely bound to bridging oxygens in the silicate network at the SiO2 grain surface. PMID- 16375278 TI - Low temperature properties of acetonitrile confined in MCM-41. AB - The effect of confinement on the phase changes and dynamics of acetonitrile in mesoporous MCM-41 was studied by use of adsorption, FT-IR, DSC, and quasi-elastic neutron scattering (QENS) measurements. Acetonitrile molecules in a monolayer interact strongly with surface hydroxyls to be registered and perturb the triple bond in the C[triple bond]N group. Adsorbed molecules above the monolayer through to the central part of the cylindrical pores are capillary condensed molecules (cc-acetonitrile), but they do not show the hysteresis loop in adsorption desorption isotherms, i.e., second order capillary condensation. FT-IR measurements indicated that the condensed phase is very similar to the bulk liquid. The cc-acetonitrile freezes at temperatures that depend on the pore size of the MCM-41 down to 29.1 A (C14), below which it is not frozen. In addition, phase changes between alpha-type and beta-type acetonitriles were observed below the melting points. Application of the Gibbs-Thomson equation, assuming the unfrozen layer thickness to be 0.7 nm, gave the interface free energy differences between the interfaces, i.e., Deltagamma(l/alpha) = 22.4 mJ m(-2) for the liquid/pore surface (ps) and alpha-type/ps, and Deltagamma(alpha/beta) = 3.17 mJ m(-2) for alpha-type/ps and beta-type/ps, respectively. QENS experiments substantiate the differing behaviors of monolayer acetonitrile and cc acetonitrile. The monolayer acetonitrile molecules are anchored so as not to translate. The two Lorentzian analysis of QENS spectra for cc-acetonitriles showed translational motion but markedly slowed. However, the activation energy for cc-acetonitrile in MCM-41 (C18) is 7.0 kJ mol(-1) compared to the bulk value of 12.7 kJ mol(-1). The relaxation times for tumbling rotational diffusion of cc acetonitrile are similar to bulk values. PMID- 16375279 TI - Dispersion of acid-treated carbon nanofibers into gel matrices prepared by the sol-gel method. AB - 1-Naphthol has been used as an in-situ fluorescent probe to characterize the dispersibility of carbon nanofibers (CNFs) into the sol-gel matrix of silicon alkoxide. The ion-pair fluorescence of 1-naphthol was found in the gel dispersing acid-treated CNFs instead of 1Lb fluorescence, which was preferred in the low polar gel matrix. This indicates that 1-naphthol easily interacts with oxidized groups present on the surface of the acid-treated CNFs due to the high dispersibility of the CNFs into the gel matrix. The oxidized groups on the CNF surface are useful for preventing self-assembly and/or aggregation of the CNFs in the gel matrix. PMID- 16375280 TI - Probing structure in the polymorphic domain of the L-enantiomer of N-benzoyl phenylalanine by means of 2D solid-state NMR spectroscopy and DFT calculations. AB - A study of polymorphism using a range of solid-state NMR techniques is presented. We demonstrate the existence of at least six polymorphs in a sample of N-benzoyl L-phenylalanine. We also present methodology for the characterization of the protonation state, hydrogen bonding, and molecular conformation for the polymorphs, together with results of such a characterization for one of the polymorphs present in our sample. DFT modeling is used to investigate the separate effects hydrogen bonding and molecular conformation have on the chemical shift tensor. PMID- 16375281 TI - Effect of a coadsorbent on the performance of dye-sensitized TiO2 solar cells: shielding versus band-edge movement. AB - The mechanism by which the adsorbent chenodeoxycholate, cografted with a sensitizer onto TiO2 nanocrystals, alters the open-circuit photovoltage and short circuit current of dye-sensitized solar cells was investigated. The influence of tetrabutylammonium chenodeoxycholate on dye loading was studied under a variety of conditions in which the TiO2 films were exposed to the sensitizing dye and coadsorbent. Photocurrent--voltage measurements combined with desorption studies revealed that adding chenodeoxycholate reduces the dye loading by as much as 60% while having a relatively small effect on the short-circuit photocurrent. Calculations along with measurements showed that even at low loading, enough dye is present to absorb a significant fraction of the incident light in the visible spectrum. In concurrence with the observations of others, we find evidence for weakly and strongly adsorbed forms of the dye resulting from either different binding conformations or aggregates. The most strongly adsorbed dyes are less susceptible to displacement by chenodeoxycholate than those that are weakly adsorbed. While having no observable effect on dye coverage, multiple exposures of a TiO2 film to a dye solution substantially increased the fraction of strongly adsorbed dye as judged by the resistance of the adsorbed dye to displacement by chenodeoxycholate. Measurements of the open-circuit voltage as a function of the photocharge density, determined by infrared transmittance, showed that chenodeoxycholate not only shifts the conduction band edge to negative potentials, but also significantly increases the rate of recombination. The net effect of adding chenodeoxycholate is, however, to improve the photovoltage. PMID- 16375282 TI - Oxygen reduction at platinum monolayer islands deposited on Au(111). AB - Atomic monolayer islands of Pt, namely, two-dimensional Pt nanoparticles, on a Au(111) electrode have been studied for the first time, focusing on their electrocatalytic activities for oxygen reduction in acid solutions. The Pt islands' electrodes were prepared using the self-assembled technique of thiols together with the replacement of Pt with a Cu monolayer. The states of adsorbed OH and the catalytic activities of oxygen reduction were sensitive to the Pt island size. As island size decreased, a delay in the reduction of surface oxide was observed. However, negligible influence of adsorbed OH on activity for oxygen reduction was observed. Pt islands of sizes ranging from 5 to 10 nm showed higher specific catalytic activities for oxygen reduction. Specific catalytic activities decreased by a factor of 10 with a decrease in island sizes from 5.5 to 3.1 nm. Size effects observed in Pt monolayer islands were discussed in comparison with three-dimensional nanoparticles, to obtain information concerning the size effects of metal nanoparticles. PMID- 16375283 TI - Prediction of the formation and stabilities of energetic salts and ionic liquids based on ab initio electronic structure calculations. AB - A computational approach to predict the thermodynamics for forming a variety of imidazolium-based salts and ionic liquids from typical starting materials is described. The gas-phase proton and methyl cation acidities of several protonating and methylating agents, as well as the proton and methyl cation affinities of many important methyl-, nitro-, and cyano-substituted imidazoles, have been calculated reliably by using the computationally feasible DFT (B3LYP) and MP2 (extrapolated to the complete basis set limit) methods. These accurately calculated proton and methyl cation affinities of neutrals and anions are used in conjunction with an empirical approach based on molecular volumes to estimate the lattice enthalpies and entropies of ionic liquids, organic solids, and organic liquids. These quantities were used to construct a thermodynamic cycle for salt formation to reliably predict the ability to synthesize a variety of salts including ones with potentially high energetic densities. An adjustment of the gas phase thermodynamic cycle to account for solid- and liquid-phase chemistries provides the best overall assessment of salt formation and stability. This has been applied to imidazoles (the cation to be formed) with alkyl, nitro, and cyano substituents. The proton and methyl cation donors studied were as follows: HCl, HBr, HI, (HO)2SO2, HSO3CF3 (TfOH), and HSO3(C6H4)CH3 (TsOH); CH3Cl, CH3Br, CH3I, (CH3O)2SO2, CH3SO3CF3 (TfOCH3), and CH3SO3(C6H4)CH3 (TsOCH3). As substitution of the cation with electron-withdrawing groups increases, the triflate reagents appear to be the best overall choice as protonating and methylating agents. Even stronger alkylating agents should be considered to enhance the chances of synthetic success. When using the enthalpies of reaction for the gas-phase reactants (eq 6) to form a salt, a cutoff value of -13 kcal mol(-1) or lower (more negative) should be used as the minimum value for predicting whether a salt can be synthesized. PMID- 16375284 TI - Thermodynamic and dielectric studies concerning the influence of cylindrical submicrometer confinement on heptyloxycyanobiphenyl. AB - Measurements of the specific heat and the static dielectric permittivity of heptyloxycyanobiphenyl (7OCB) confined to the 0.2 microm diameter parallel cylindrical pores of Anopore membranes in the isotropic phase and nematic mesophase, are presented. A comparison between the bulk and the confined 7OCB in treated and untreated pore wall surfaces using a chemical surfactant (HTBA) is performed. Both the treated and untreated membrane confinements seem to affect the nematic-to-isotropic phase transition by a downshift in transition temperature and some rounding at the specific-heat maximum, in a way similar to that which was earlier published for other liquid crystals confined in the same geometry. The static dielectric measurements clearly point out that untreated membrane confinement is axial, with the nematic director aligned parallel to the pore axis being homeotropic bulklike, i.e., with the nematic director aligned perpendicular to the electrode cell surfaces. After chemical surfactant treatment, the nematic director is constrained in a radial alignment being perpendicular to the pore walls. The dielectric measurements are revealed to be specially sensible to analyze the surface-induced nematic order due to the pore wall. The tricritical nature of the nematic-to-isotropic phase transition in bulk 7OCB as well as in treated and untreated Anopore confined geometries is discussed through both the specific heat and the static dielectric data. PMID- 16375285 TI - Thermolysis of fluorinated single-walled carbon nanotubes: identification of gaseous decomposition products by matrix isolation infrared spectroscopy. AB - The thermal decomposition of fluorinated single-walled carbon nanotubes (F SWNTs), known to result in pristine SWNTs, has been investigated by freezing the gaseous products formed at temperatures between 50 and 500 degrees C under high vacuum in an argon matrix at 10-20 K and analyzing the trapped species by IR spectroscopy. The major products of F-SWNT decomposition are carbonyl fluoride (COF2) below 300 degrees C and CF4 above 300 degrees C. For comparison, graphite fluoride is stable thermally up to 300 degrees C under these conditions, and the major gas-phase species at temperatures below 500 degrees C are CF4 and the CF3 radical. F-SWNTs are thermally less stable than graphite fluoride, and etching of the nanotubes is observed at lower thermolysis temperatures. PMID- 16375286 TI - Unusual dependence of particle architecture on surfactant concentration in partially fluorinated decylpyridinium templated silica. AB - A series of porous silica particles is prepared with different concentrations of the fluorinated cationic surfactant 1-(3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10) heptadecafluorodecyl)pyridinium chloride (HFDePC) to trace the changes in pore structure and particle morphology as the surfactant concentration increases. At the lowest concentration studied (1.5 mmol/L), the product consists of small round particles with close-packed cylindrical mesopores. As the HFDePC concentration increases, macroporous voids are introduced to create multi chambered hollow particles with mesoporous walls. With a still higher concentration of HFDePC the macropore volume decreases, and elongated, tactoid like nanoparticles are formed with random mesh-phase pores oriented with silica layers perpendicular to the main axis of the particles. Further increasing the concentration of HFDePC eventually leads to the formation of round particles with disordered pores. These changes are consistent with increasing HFDePC concentration favoring increasingly oblate or disklike micelles. The process of forming the elongated particles with random mesh-phase structure is investigated by TEM of chilled and dried samples. The results indicate that the oriented tactoid-like structure forms spontaneously within 2 min by co-assembly of silica and HFDePC rather than by preferred growth perpendicular to the layers. The particle shape and layer orientation are consistent with what would be expected for a liquid-crystal particle with orientation-dependent surface tension. Finally, we compare samples prepared with a high HFDePC and with good or poor mixing. With inadequate mixing, a gel layer forms at the top of the sample which is composed of elongated mesoporous particles with a thick coating of microporous silica. The lower particulate phase contains small disordered particles similar to those obtained in a well-mixed sample. Presumably, the structure of the upper layer results from initial immiscibility of the precursor and slow diffusion of silicates out of the gel. PMID- 16375287 TI - Highly ordered self-assembly with large area of Fe3O4 nanoparticles and the magnetic properties. AB - Monodisperse Fe3O4 nanoparticles (NPs) with narrow size distribution are synthesized by a high-temperature solution-phase method. The diameter of the as synthesized NPs is tuned from 2 to 14 nm by varying the reaction conditions. Highly ordered superlattice structures of the Fe3O4 NPs with areas extending over 0.8 microm x 0.7 microm have been successfully obtained. The magnetic properties are investigated in their different states, such as in the solid state and diluted in wax with different concentrations. Some magnetic properties enhanced by increasing interparticle distances, such as the remanent magnetization and coercive field at low temperature, were noticed. Furthermore, we also observed that the saturation magnetization changed with temperature as expected. The preliminary explanation for the properties mentioned above is proposed. PMID- 16375288 TI - Relaxation and short time dynamics of bulk liquids and fluids confined in spherical cavities and slit pores. AB - The density of states for bulk and confined fluids have been modeled using a recently proposed gamma distribution (Krishnan, S. H.; Ayappa, K. G. J. Chem. Phys. 2004, 121, 3197). The gamma distribution results in a closed form analytical expression for the velocity autocorrelation function and the relaxation time of the fluid. The two parameters of the gamma distribution are related analytically to the second and fourth frequency moments of the fluid using short time expansions. The predictions by the proposed gamma model are compared with the velocity autocorrelation functions obtained using the theory of instantaneous normal modes (INMs) and from molecular dynamics simulations. The model is applied to a bulk soft sphere liquid and fluids confined in a spherical cavity and slit-shaped pores. The gamma model is able to capture the resulting changes in relaxation time due to changes in density and temperature extremely well for both the bulk liquid and confined inhomogeneous fluid situations. In all cases, the predictions by the gamma model are superior to those obtained from the INM theory. In the case of the fluid confined in a slit pore, the loadings were obtained from a grand canonical Monte Carlo simulation where the pore is equilibrated with a bulk fluid. This is similar to a confinement situation in a surface force apparatus. The predicted relaxation times vs pore widths from the gamma model are seen to accurately capture the oscillations due to formation and disruption of layers within the slit pore. PMID- 16375289 TI - Novel mesoporous mixed Nb-M (M = V, Mo, and Sb) oxides for oxidative dehydrogenation of propane. AB - Novel thermally stable mesoporous mixed metal Nb-M (M = V, Mo and Sb) oxides were synthesized in the presence of a nonionic Pluronic P123 surfactant. These oxides displayed promising pore structures and chemical compositions for selective oxidative functionalization of propane: high surface areas (up to 200 m2/g), large pore sizes (5-14 nm), and high pore volumes (up to 0.46 cm3/g). The oxidative dehydrogenation of propane to propylene over mesoporous mixed metal Nb M oxides employed as a probe reaction suggested that the M component was dispersed as the molecular surface species and also formed a solid solution with NbOx in the inorganic walls of these mesoporous mixed metal oxides. PMID- 16375290 TI - Diluent effects on the Debye-type dielectric relaxation in viscous monohydroxy alcohols. AB - With the recognition that the Debye-type dielectric relaxation of liquid monohydroxy alcohols does not reflect the structural relaxation dynamics associated with the viscous flow and the glass transition, its behavior upon dilution is expected to differ from that of real alpha-processes. We have investigated the Debye-type dielectric relaxation of binary alcohol/alkane mixtures across the entire concentration range in the supercooled regimes. The focus is on 2-ethyl-1-hexanol in two nonpolar liquids, 3-methylpentane and squalane, which are more fluid and more viscous than the alcohol, respectively. The Debye relaxation is found to occur only for alcohol mole fractions x > 0.2 and is always accompanied by a non-Debye relaxation originating from the alcohol component. Prior to its complete disappearance, the Debye relaxation is subject to broadening. We observe that the Debye dynamics of 2-ethyl-1-hexanol is accelerated in the more fluid 3-methylpentane, while the more viscous squalane leads to longer Debye relaxation times. The present experiments also provide evidence that the breakdown of the Debye relaxation amplitude does not imply the absence of hydrogen-bonded structures. PMID- 16375291 TI - Adsorption and structural properties of ordered mesoporous carbons synthesized by using various carbon precursors and ordered siliceous P6mm and Ia3d mesostructures as templates. AB - Adsorption and structural properties of inverse carbon replicas of two ordered siliceous P6mm and Ia3d mesostructures have been studied by nitrogen adsorption, powder X-ray diffraction, and transmission electron microscopy. These carbon replicas were prepared by filling the pores of SBA-15 and KIT-6 siliceous templates with various carbon precursors followed by carbonization and silica dissolution. Sucrose, furfuryl alcohol, acenaphthene, mesophase pitch, and petroleum pitch were used to obtain inverse carbon replicas of SBA-15 and KIT-6. While structural properties of the resulting ordered mesoporous carbons are mainly determined by the hard template used, their adsorption properties depend on the type of the carbon precursor. PMID- 16375292 TI - Molten salt route toward the growth of ZnO nanowires in unusual growth directions. AB - ZnO nanowires with unusual growth directions, such as the approximate 102 and the 100 directions, were prepared by using the LiCl molten salt synthetic method. Intrinsic crystallographic structures and the growth directions of the as prepared ZnO nanowires were investigated by using selected area electron diffraction and high-resolution transmission electron microscopy. In the present case, Li+ and Cl- ions of molten salts may bind with O2- and Zn2+ ions, respectively, of the {101} and {001} polar surfaces of the ZnO crystals, resulting in the decrease of their surface energies and tuning the growth directions by blocking the growth on the polar surfaces. A combination of the growth along the <102>, <100>, and <210> directions may lead to the formation of complex tree like ZnO dendrites. Strong green light emission was observed from room-temperature PL spectra of the as-prepared ZnO nanowires. This molten-salt synthetic process could be extended to synthesize other kinds of unusual 1D nanomaterials with specific crystal structures and properties. PMID- 16375293 TI - Preparation by high-energy milling, characterization, and catalytic properties of nanocrystalline TiO2. AB - Titanium dioxide (TiO2) is widely used for applications in heterogeneous photocatalysis. We prepared nanocrystalline powders of the anatase as well as the rutile modification by high-energy ball milling of the coarse grained source materials for up to 4 h. The resulting average grain size was about 20 nm. The morphology of the powders was investigated with transmission electron microscopy, X-ray powder diffraction, and BET surface area determination. Measurements of the catalytic activity reveal a maximum as a function of the milling time at about 40 min. This maximum could be explained by a superposition of two counteracting effects. The first one is the increase of the specific surface area resulting in an increase of the catalytic activity, and the second one is a change of the electronic structure at the surface of the TiO2 particles corresponding to a reduction of the surface. The latter one was confirmed by light absorption experiments, X-ray photoelectron spectroscopy, and electron paramagnetic resonance spectroscopy. PMID- 16375294 TI - Synthesis of manganese oxide electrodes with interconnected nanowire structure as an anode material for rechargeable lithium ion batteries. AB - Manganese oxide electrodes composed of interconnected nanowires are electrochemically synthesized in manganous acetate solution at room temperature without any template and catalyst. Annealing temperature affects the electrode morphology, crystallization, and electrochemical performance. Scanning electron microscope (SEM) results show that nanowires are uniformly distributed and sizes are about 12-18 nm in diameter; the diameter decreases to about 8-12 nm after annealing at 300 degrees C. X-ray diffraction (XRD) and transmission electron microscope (TEM) images indicate that nanowires have poor crystalline characteristics. The higher the annealing temperature, the higher the crystalline degree is in manganese oxide. The synthesized anode material shows a much larger capacity than the traditional graphite materials for lithium storage. After annealing at 300 degrees C, the electrode's reversible capacity reaches 800 mAhg( 1), and the specific capacity retention remains nearly constant after 100 cycles. PMID- 16375295 TI - Molecular motion of isolated linear alkanes in nanochannels. AB - The mobility of a series of linear alkanes in their inclusion compound with tris(o-phenylenedioxy)spirotriphosphazene is studied by high-resolution carbon proton magic-angle spinning solid-state NMR spectroscopy. Two different carbon proton dipolar recoupling experiments are compared with respect to their ability to yield precise site-specific, motion-averaged dipolar coupling constants. The most accurate results are obtained by analysis of extrema positions in Lee Goldburg cross-polarization build-up curves. We present a comprehensive collection of coupling constants, which evidence a rotational motion of the all trans chains around the channel axis, with some further averaging due to additional fluctuations, as previously found for alkanes in other host matrices such as urea. The order parameter increases toward the inner parts of the chains, and is largely independent of chain length. Notably, chains in a TPP host are not more ordered than in urea, even though the average TPP channel diameter is reported to be smaller. Significantly decreased order is found for highly filled short-alkane samples, which is interpreted in terms of an increased rate of mutual collisions. From residual dipolar couplings as well as carbon chemical shifts, we derive similar amounts of gauche conformers. Translational motions along the channels are further studied by proton double-quantum spectroscopy, which probes guest-host dipolar couplings. The extent of local-scale lateral motion is again correlated with the sample filling, and is a weak function of temperature, as expected from a case in which highly restricted single-file diffusion should dominate the mobility. Characteristic effects of sample aging are apparent in all our experiments. PMID- 16375296 TI - Preparation and nonlinear optical properties of inorganic-organic hybrid films with various substituents on chromophores. AB - Four kinds of hydroxy-ended azobenzene-type chromophores containing different substituent groups as electron donor or electron acceptor were synthesized and further reacted with 3-isocyanatopropyltriethoxysilane (ICTES) to give alkoxysilane dyes via a urethane reaction. Following a sol-gel process of the alkoxysilane dyes, the inorganic-organic hybrid nonlinear optical (NLO) films were successfully prepared. Molecular structures of the resultant films were confirmed by elemental analysis, FTIR, and 1H NMR. The betaCT mu(g) values of the chromophores were evaluated by a solvatochromic method, and the second harmonic coefficients (d33) of the hybrid films were measured by in situ second harmonic generation (SHG) measurement. The hybrid films exhibited large optical nonlinearity and full transparency in the visible range. The effects of substituent group and position on betaCT mu(g) values of the chromophores and d33 values of the films were also discussed. PMID- 16375297 TI - Electrochemical growth of nickel hollow nanostructures on copper substrates. AB - We have shown hollow Ni nanonodules with outer diameters of 80-200 nm and wall thicknesses of 5-25 nm could be prepared by electrochemical deposition in the NiCl2 + dimethyl sulfoxide + C2H4O3 + H2O system, and the products were high purity. In particular, Ni hollow nanonodule structures or highly assembling Ni hollow nanostructures can be selected by varying the compositions of the solvent mixture. Apart from the hollow grain size, the wall thickness can also easily be controlled by varying the electrochemical parameters, salt concentration, and deposition time. The typical coercivity of Ni hollow nanostructures with particle sizes of about 100-150 nm was much bigger than that of the bulk Ni. PMID- 16375298 TI - Chemiluminescence of CdTe nanocrystals induced by direct chemical oxidation and its size-dependent and surfactant-sensitized effect. AB - CdTe nanocrystals (NCs) capped with thioglycolic acid (TGA) were synthesized via a microwave-assisted method. The chemiluminescence (CL) of CdTe NCs induced by directly chemical oxidation and its size-depended and surfactant-sensitized effect in aqueous solution were then investigated. It was found that oxidants, especially hydrogen peroxide and potassium permanganate, could directly oxidize CdTe NCs to produce strong CL emission in basic conditions. The oxidized CL of CdTe NCs displayed size-dependent effect and its intensity increased along with increasing the sizes of the NCs. Moreover, the CL intensity could, if surfactants CTAB or beta-cyclodextrin were added to the above CL system, be sensitized to some degree. The sensitized CL induced by CTAB and beta-cyclodextrin is mainly contributing to the formation of aggregate nanostructure and the micellar micronanoenvironment, respectively. The possible oxidized CL mechanisms were further examined by means of photoluminescence spectra, CL spectra, and transmission electron microscopy studies. The CL properties of CdTe NCs not only will be helpful to study physical chemistry properties of semiconductor nanocrystals but also are expected to find use in many fields such as luminescence devices, bioanalysis, and multicolor labeling probes. PMID- 16375299 TI - High-quality ultralong Sb2S3 nanoribbons on large scale. AB - Large-scale, ultralong, single-crystalline Sb2S3 nanoribbons were prepared by directly reacting SbCl3 and Na2S2O3 solutions, without any organics used in the experiment. The nanoribbons were analyzed by a range of methods. The nanoribbons are usually several millimeters in length, typically 200-500 nm in width and 30 80 nm in thickness. The structure of the nanoribbons is determined to be of the orthorhombic phase. The growth mechanism of the nanoribbons was investigated based on high-resolution transmission electron microscopy observations. Optical absorption experiment shows that the nanoribbon is a semiconductor with a bandwidth Eg approximately 1.5 eV, near to the optimum for photovoltaic conversion, suggesting that Sb2S3 nanoribbons could be used in solar energy and photoelectronic applications. PMID- 16375300 TI - Self-oscillating soluble-insoluble changes of a polymer chain including an oxidizing agent induced by the Belousov-Zhabotinsky reaction. AB - A new type of self-oscillating polymer was prepared by utilizing the Belousov Zhabotinsky reaction. In this study, capture sites with a positive charge for an oxidizing agent as a counterion were incorporated into the copolymer of N isopropylacrylamide and the ruthenium complex as a catalyst. Soluble-insoluble self-oscillation of the polymer was first achieved without adding an oxidizing agent. The effect of temperature on the self-oscillating behavior was investigated. It was clarified that the polymer had two advantageous characteristics because of the higher LCST; one is to enable self-oscillation around body temperature, and the other is to cause the oscillation for a longer time without intermolecular aggregation among the polymer chains in the reduced state. This achievement of self-oscillation of polymer chains including an oxidizing agent may lead to their practical use under oxidant-free conditions. PMID- 16375301 TI - Photoluminescence properties and analysis of spectral structure of Eu3+-doped SrY2O4. AB - The aim of this work is to report on the luminescence properties of SrY2O4 activated by Eu3+ ion. Powder samples were prepared by solid-state reaction. X ray diffraction powder data, photoluminescence, and high-resolution spectroscopy were carried out. Results revealed that the Eu3+ ions occupied three nonequivalent sites, with one at the Sr site, one at the Y(1) site, and another at the Y(2) site. Their spectra wavelengths for the 7F0-5D0 transition are located at 578.49, 581.86, and 580.63 nm, respectively. The corresponding charge transfer transitions are located at 248, 257, and 270 nm, respectively, which are also confirmed by theoretical analysis. PMID- 16375302 TI - Controllable template synthesis of Ni/Cu nanocable and Ni nanotube arrays: a one step coelectrodeposition and electrochemical etching method. AB - We describe a new synthetic approach to fabricate Ni/Cu nanocable arrays by co depositing nickel and copper atoms into the pores of anodic alumina membranes and to fabricate Ni nanotube arrays by selectively etching the Cu cores from the Ni/Cu nanocable arrays. The formation of the Ni-shelled Ni/Cu nanocables is attributed to the Ni ions adsorbed on the pore walls by a chemical complexation through hydroxyl groups. By varying electrodepositon parameters in this technique, we can control the lengths of nanocables and nanotubes, the shell thickness of the nanocables, and the wall thickness and surface morphology of the nanotubes. PMID- 16375303 TI - Charging effects in a CdSe nanotetrapod. AB - The charging effects in a CdSe nanotetrapod have been theoretically investigated by using an atomistic pseudopotential method. We showed that the simple quasiparticle equation based on classical electrostatic consideration can be derived from the many-body GW equation under proper approximations. We found that the surface polarization potential can significantly change the electron wave functions, and there is an incomplete cancellation for this potential between the single particle energies and the electron-hole Coulomb interaction. Thus, it is necessary to include this potential in the calculation for complex unconvex systems. We also calculated the electron addition energies for a tetrapod. Unlike a simple spherical quantum dot, in which the addition energies are almost a constant, there is a large variation in the calculated addition energies for different numbers of electrons in a tetrapod. PMID- 16375304 TI - Uniform gold nanorod arrays from polyethylenimine-coated alumina templates. AB - Monolithic Au nanorod arrays can be grown by electrodeposition in Au-backed nanoporous alumina templates using polyethylenimine (PEI) as an adhesion layer, with excellent height control between 300 nm and 1.4 microm. The local height distribution can be extremely narrow with relative standard deviations well below 2%. The uniform growth rate appears to be determined by the adsorbed PEI matrix, which controls the growth kinetics of the grains comprising the nanorods. The nanorods can be retained as free-standing 2D arrays after careful removal of the AAO template. Reflectance spectroscopy reveals a collective plasmon mode with a maximum near 1.2 microm, in accord with recent calculations for 2D arrays of closely spaced cylindrical nanoparticles. PMID- 16375305 TI - Structural and dynamic aspects of hydrogen-bonded complexes and inclusion compounds containing alpha,omega-dicyanoalkanes and urea, investigated by solid state 13C and 2H NMR techniques. AB - Solid-state 13C NMR and 2H NMR techniques have been used to investigate structural and dynamic properties of the 1,4-dicyanobutane/urea and 1,5 dicyanopentane/urea 1:1 hydrogen-bonded complexes and the 1,6-dicyanohexane/urea inclusion compound. The pure crystalline phase of urea has also been investigated. The 13C NMR studies have focused on 13C chemical shift anisotropy and second-order quadrupolar effects (arising from 13C-14N interaction) for the urea molecules and the cyano groups of the alpha,omega-dicyanoalkanes. Parameters describing these interactions are derived and are discussed in relation to the known structural properties of these materials. Comparison of 13C chemical shift anisotropies of the cyano carbons and rates of 13C dipolar dephasing suggest that 1,4-dicyanobutane and 1,5-dicyanopentane are effectively static, whereas 1,6 dicyanohexane has greater mobility. 2H NMR line shape analysis for the 1,4 dicyanobutane/urea-d4 and 1,5-dicyanopentane/urea-d4 complexes indicates that the only motion of the urea molecules that is effective on the 2H NMR time scale is a rapid libration about the C=O bond over an angular range of about 26 degrees . For the 1,6-dicyanohexane/urea-d4 inclusion compound, the 2H NMR line shape is consistent with a motion comprising 180 degrees jumps about the C=O bond at rates that are intermediate on the 2H NMR time scale. In addition, rapid libration about the C=O bond also occurs over an angular range of about 20 degrees . The dynamic properties of the urea molecules in these materials are compared with those of urea molecules in other crystalline environments. PMID- 16375306 TI - How does surface modification aid in the dispersion of carbon nanofibers? AB - Small-angle light scattering is used to assess the dispersion behavior of vapor grown carbon nanofibers suspended in water. These data provide the first insights into the mechanism by which surface treatment promotes dispersion. Both acid treated and untreated nanofibers exhibit hierarchical morphology consisting of small-scale aggregates (small bundles) that agglomerate to form fractal clusters that eventually precipitate. Although the morphology of the aggregates and agglomerates is nearly independent of surface treatment, their time evolution is quite different. The time evolution of the small-scale bundles is studied by extracting the size distribution from the angle-dependence of the scattered intensity, using the maximum entropy method in conjunction with a simplified tube form factor. The bundles consist of multiple tubes possibly aggregated side-by side. Acid oxidation has little effect on this bundle morphology. Rather acid treatment inhibits agglomeration of the bundles. The time evolution of agglomeration is followed by fitting the scattering data to a generalized fractal model. Agglomerates appear immediately after cessation of sonication for untreated fibers but only after hours for treated fibers. Eventually, however, both systems precipitate. PMID- 16375307 TI - Effect of the tube diameter distribution on the high-temperature structural modification of bundled single-walled carbon nanotubes. AB - We present results of a systematic high-resolution transmission electron microscopy study of the thermal evolution of bundled single-walled carbon nanotubes (SWNTs) subjected to approximately 4-h high-temperature heat treatment (HTT) in a vacuum at successively higher temperatures up to 2200 degrees C. We have examined purified SWNT material derived from the HiPCO and ARC processes. These samples were found to thermally evolve along very different pathways that we propose depend on three factors: (1) initial diameter distribution, (2) concomitant tightness of the packing of the tubes in a bundle, and (3) the bundle size. Graphitic nanoribbons (GNR) were found to be the dominant high-temperature filament in ARC material after HTT = 2000 degrees C; they were not observed in any heat-treated HiPCO material. The first two major steps in the thermal evolution of HiPCO and ARC material agree with the literature, i.e., coalescence followed by the formation of multiwall carbon nanotubes (MWNTs). However, ARC material evolves to bundled MWNTs, while HiPCO evolves to isolated MWNTs. In ARC material, we find that the MWNTs collapse into multishell GNRs. The thermal evolution of these carbon systems is discussed in terms of the diameter distribution, nanotube coalescence pathways, C-C bond rearrangement, diffusion of carbon and subsequent island formation, as well as the nanotube collapse driven by van der Waals forces. PMID- 16375308 TI - Study on the formation of the ketonic defects in the thermal degradation of ladder-type poly(p-phenylenes) by vibrational spectroscopy. AB - We have investigated the thermal degradation in air by Fourier transform infrared spectroscopy of a ladder-type copolymer containing fluorene units in the backbone (Me-LPF), to reveal the formation of the ketonic defects. As thermal treatment of Me-LPF film at 200 degrees C in air proceeds, a new group of complex absorption bands due to degradation products arises in the range between 1800 and 1600 cm( 1). The observed overlapping bands were separated and assigned by utilizing the second-derivative IR spectral analysis, which can narrow the peak width to one third of the originals and thereby eases the analysis. The degraded products were assigned as fluorenone (1718 cm(-1)) and benzophenone (Ar-(C=O)-Ar) (1665 cm( 1)), formed by the oxidation of the backbone, and acylphenone (Ar-(C=O)-R) (1685 cm(-1)) from the side chain. The fluorenone was found to be the major component among the degraded products in the main chain, and the time and temperature dependence indicated that the oxidation is a kind of autocatalytic radical-chain process. The oxidation can reach a very high degree (approximately 30% for 6 h oxidation at 240 degrees C estimated by absorption of the alkyl). Our results suggest the possibility of the oxidation of the 9-bialkylfluorene sites. We propose that the degradation of the alkyl in the side chain can help the radicals to propagate in the chain reaction. PMID- 16375309 TI - Silver nanodisks: optical properties study using the discrete dipole approximation method. AB - The simulated optical properties of silver nanodisks are presented. The extinction, absorption, and scattering efficiencies are calculated using the discrete dipole approximation. The influence of the nanodisk size, truncature (snip), aspect ratio, and environment on the plasmon resonance bands is investigated. In particular, the dipolar and multipolar resonance peak positions have been related to the specific features of the nanodisk geometry. An interpretation of the origins of each multipolar mode is proposed for the first time taking into account this geometry. PMID- 16375310 TI - Formation and characterization of crystalline molecular arrays of gas molecules in a 1-dimensional ultramicropore of a porous copper coordination polymer. AB - Molecules and atoms confined in a nanospace may have properties distinctly different from those of the bulk fluid, owing to the formation of a specific molecular array characteristic of nanospace. In situ synchrotron powder X-ray diffraction measurements have been used to observe confined guest molecules such as N2, O2, Ar, and CH4 in the well-regulated ultramicropore of a copper coordination polymer, 1 ([Cu2(pzdc)2pyz]: pzdc = 2,3-pyrazinedicarboxylate and pyz = pyrazine). The obtained crystal structures indicate that guest molecules are confined in a linear fashion to form crystalline-like regular ordered arrays, in contrast to the situation in the gas and liquid state, even at temperatures above the boiling point, and the ordered arrays are characteristic of the kind of gas molecule and the geometrical and potential properties of the ultramicropore of 1. PMID- 16375311 TI - The influence of defects on the morphology of Si (111) etched in NH4F. AB - We have implemented a kinetic Monte Carlo (KMC) simulation to study the effects of wafer miscut and wafer defects on the morphologies of Si (111) surfaces etched in NH4F. Although a conventional KMC simulation reproduced previously published results, it failed to produce the morphologies observed in our experiments. By introducing both dopant sites and lattice defect sites into the model, we are able to simulate samples having different dopant elements and densities as well as different defect concentrations. Using the modified KMC simulation, the simulated surface morphologies agree well with the morphologies observed in our experiments. The enhanced model also gives insights to the formation mechanism for multiple level stacking pits, a notable morphology on the etched surfaces of samples with very small miscut angles. PMID- 16375312 TI - A model for the structure of MCM-41 incorporating surface roughness. AB - The surface area of MCM-41 mesoporous silica, estimated by several models in the literature, is significantly less than the value derived from BET analysis of nitrogen adsorption at 77.4 K. In the past, the difference has been attributed to several reasons including the errors involved in the BET analysis of the multilayer-capillary condensation region and the heterogeneity of the walls. In the present work, we present an alternate model of MCM-41 based on molecular simulations that gives surface area values that are in closer agreement to those determined by experiment. The model incorporates bulk heterogeneity of the material, surface hydroxyls, and most importantly, physical deformations or indentations of the pore surface. The model predicts small-angle X-ray diffraction (XRD) and wide-angle X-ray scattering (WAXS) results that are consistent with experimental data as well as surface areas and pore volumes that compare favorably with published experimental results. The simulation results are consistent with the hypothesis that the interstitial space in MCM-41 is relatively amorphous despite the regular arrangement of the mesopores. The surface roughness associated with the amorphous structure increases the surface area beyond the nominal value produced by assuming smooth cylindrical pores. PMID- 16375313 TI - Effects of chain length and heat treatment on the nanotribology of alkylsilane monolayers self-assembled on a rough aluminum surface. AB - The conformational order of alkylsilane monolayers self-assembled on a rough aluminum surface is affected by the molecular chain length and the thermal history of the sample. These monolayers have been characterized by grazing angle FTIR spectroscopy. Tribological mechanisms were explored using initial molecular conformation order, sliding distance, normal load, and substrate compliance as experimental variables. Results indicate that the initial conformational disorder of the molecules determines the level of friction at the commencement of sliding. Adverse changes in dynamic friction and monolayer life during sliding are not thermally induced but are related to substrate roughness and local plasticity. Plastic deformation reduces the spatial density of the alkylsilane monolayer and is accentuated by an increase in the normal load. PMID- 16375314 TI - Entropically mediated polyolefin blend segregation at buried sapphire and air interfaces investigated by infrared-visible sum frequency generation vibrational spectroscopy. AB - The segregation behavior of binary polymer blends at hydrophilic solid sapphire and air interfaces was investigated by infrared-visible sum frequency generation (SFG) vibrational spectroscopy. SFG spectra were collected from a bulk miscible blend consisting of identical molecular weight (approximately 54,000) and similar surface free energy (29-35 dyn/cm) components of atactic polypropylene (aPP) and aspecific poly(ethylene-co-propylene) rubber (aEPR). Characteristic CH resonances of the blend were contrasted with those of the individual components at both buried (sapphire/polymer) and free (air/polymer) interfaces. Preferential segregation of the aPP component was observed after annealing at both air/polymer and sapphire/polymer interfaces. SFG spectra revealed ordering of the polymer backbone segments with the methylene (CH2) groups perpendicular to the surface at the sapphire interface and the methyl (CH3) groups upright at the air interface. The SFG results indicate that the surface composition can be determined from the peak intensities that are characteristic of each component and that conformational entropy played a likely role in surface segregation. aPP occupied a smaller free volume at the surface because of a statistically smaller segment length (aPP is more flexible and has a shorter length). In addition, the high density of the ordered CH3 side branches enhanced the surface activity by allowing the long-chain backbone segments of aPP to order at the surface. PMID- 16375315 TI - A study of the redox properties of MoOx/SiO2. AB - A sample of MoOx/SiO2, in which all of the Mo cations are present as isolated mono-oxo molybdate moieties, was prepared and investigated to understand the redox chemistry of such molybdate species and their ability to exchange oxygen with O2 and H2O. Raman spectroscopy was used to monitor the exchange of 18O for 16O in the Mo=O bond of isolated molybdate species, whereas mass spectrometry was used to follow the isotopic composition of the gaseous species, i.e., O2 and H2O. Reduction in H2 at 920 K results in the loss of one O atom per Mo atom, and consistent with this, all of the Mo(VI) cations are reduced to Mo(IV) cations. Raman spectroscopy shows that virtually all Mo=O bonds of the original molybdate species are lost upon reduction. While reoxidation of Mo(IV) cations by O2 is quantitative, studies using 18O2 reveal that only a small part of the newly formed Mo=O bonds are 18O labeled, and that the balance are 16O labeled, indicating that O-atom exchange between the support, SiO2, and the supported MoOx species occurs during reoxidation. Rapid exchange of O atoms was observed upon exposure of both bare SiO2 and MoOx/SiO2 to H2(18)O at 920 K, and the presence of MoOx species was found to enhance the rate of exchange. By contrast, very slow exchange of O atoms was observed when the oxidized catalyst was exposed to 18O2 at 920 K. In situ observations of the catalyst during exposure to a mixture of H2 and 18O2 at 920 K showed that all of the Mo cations remained in the VI oxidation state and that O atom exchange occurred at a rate comparable to that observed upon exposure to H2(18)O. The results of this investigation suggest that reoxidation of Mo(IV) cations following H2 reduction involves the formation of a Mo-peroxide species and subsequent O atom migration from such a species to the SiO2 support. It is proposed that the steady-state oxidation of H2 also involves the formation of Mo-peroxide species by interaction of O2 with a small number of Mo(IV) centers. The Mo-peroxide species are then rapidly reduced by H2 to form H2O and a Mo=O bond. The rapid exchange of O atoms between the gas phase and the catalyst observed during steady-state oxidation of H2 is attributed to interactions of the product H2O with the catalyst, rather than to O atom migration originating from the Mo-peroxide species formed on the catalyst surface. PMID- 16375316 TI - Effects of reduction temperature and metal-support interactions on the catalytic activity of Pt/gamma-Al2O3 and Pt/TiO2 for the oxidation of CO in the presence and absence of H2. AB - TiO2- and gamma-Al2O3-supported Pt catalysts were characterized by HRTEM, XPS, EXAFS, and in situ FTIR spectroscopy after activation at various conditions, and their catalytic properties were examined for the oxidation of CO in the absence and presence of H2 (PROX). When gamma-Al2O3 was used as the support, the catalytic, electronic, and structural properties of the Pt particles formed were not affected substantially by the pretreatment conditions. In contrast, the surface properties and catalytic activity of Pt/TiO2 were strongly influenced by the pretreatment conditions. In this case, an increase in the reduction temperature led to higher electron density on Pt, altering its chemisorptive properties, weakening the Pt-CO bonds, and increasing its activity for the oxidation of CO. The in situ FTIR data suggest that both the terminal and bridging CO species adsorbed on fully reduced Pt are active for this reaction. The high activity of Pt/TiO2 for the oxidation of CO can also be attributed to the ability of TiO2 to provide or stabilize highly reactive oxygen species at the metal-support interface. However, such species appear to be more reactive toward H2 than CO. Consequently, Pt/TiO2 shows substantially lower selectivities toward CO oxidation under PROX conditions than Pt/gamma-Al2O3. PMID- 16375317 TI - Impedance spectroscopy of water solutions: the role of ions at the liquid electrode interface. AB - We discuss the influence of the ions dissolved in a liquid on the impedance spectroscopy of a cell. Our analysis is performed in the small-voltage regime, where the actual bulk density of ions is only slightly perturbed by the external electric field. In this framework, we show that the presence of the ions can be taken into account by a surface density of charge. The agreement between the theoretical prediction, on the basis of the assumption that the ionic mobility is frequency independent, and the experimental data for the real and imaginary parts of the impedance is fairly good for frequencies larger than 100 Hz. In the low frequency range, the agreement of the theory with the experiment is rather poor. In this region, the experimental data can be successfully fitted by introducing the impedance of the metal-electrolyte interface, which is accurately represented by Zi = w(i omega)(-nu), where w and nu are two constants, with 0 < nu < 1. From the analysis of the experimental data, we determine w and nu. The theoretical predictions of our model are in good agreement with the experimental data in the investigated frequency range. PMID- 16375318 TI - Electrochemical self-assembly of alkanethiolate molecules on Ni(111) and polycrystalline Ni surfaces. AB - In this work, the electrochemical formation of alkanethiolate self-assembled monolayers (SAMs) on Ni(111) and polycrystalline Ni surfaces from alkanethiol containing aqueous 1 M NaOH solutions was studied by combining Auger electron spectroscopy (AES), X-ray photoelectron spectroscopy (XPS), electrochemical techniques, and density functional theory (DFT) calculations. Results show that alkanethiolates adsorb on Ni concurrent with NiO electroreduction. The resulting surface coverage depends on the applied potential and hydrocarbon chain length. Electrochemical and XPS data reveal that alkanethiolate electroadsorption at room temperature takes place without S-C bond scission, in contrast to previous results from gas-phase adsorption. A complete and dense monolayer, which is stable even at very high cathodic potentials (-1.5 V vs SCE), is formed for dodecanethiol. DFT calculations show that the greater stability against electrodesorption found for alkanethiolate SAMs on Ni, with respect to SAMs on Au, is somewhat related to the larger alkanethiolate adsorption energy but is mainly due to the larger barrier to interfacial electron transfer present in alkanethiolate-covered Ni. A direct consequence of this work is the possibility of using electrochemical self-assembly as a straightforward route to build stable SAMs of long-chained alkanethiolates on Ni surfaces at room temperature. PMID- 16375319 TI - Effect of the platinum content on the microstructure and micropore size distribution of Pt/alumina-pillared clays. AB - The aim of this work is to study the effect of the platinum content (0-1.8 wt % Pt) on the microstructure of an alumina-pillared clay. For this purpose, the nitrogen physisorption data at -196 degrees C, the micropore size distributions of the supported platinum catalysts, and the hydrogen chemisorption results at 30 degrees C have been analyzed and compared. The preparation of the catalysts has modified the textural properties of the Al-pillared clay support, giving rise to a loss of surface area and micropore volume. After reduction at 420 degrees C, the presence of dispersed metallic platinum with mean crystallite size in the 22 55 A range has been found by hydrogen adsorption. Comparison of all results reveals that the platinum species block the micropore entrances by steric hindrance to nitrogen access as the platinum content increases. PMID- 16375320 TI - Functionalization of carbon nanotubes via nitrogen glow discharge. AB - We have exposed single-wall carbon nanotubes (SWCNTs) to microwave-generated N2 plasma with the aim to functionalize the nanotubes. The results strongly depend on the distance between the discharge source and the sample, since nitrogen atoms generated can be lost due to recombination. No functionalization was observed when this distance was 7.0 cm. At intermediate distances (2.5 cm), the incorporation of nitrogen and oxygen onto the SWCNT was observed, while, at short distances (1 cm), products containing CN were also observed. PMID- 16375321 TI - Local electronic structure of layered Li(x)Ni0.5Mn0.5O2 and Li(x)Ni(1/3)Mn(1/3)Co(1/3)O2. AB - Samples of Li(x)Ni0.5Mn0.5O2 and Li(x)Ni(1/3)Mn(1/3)Co(1/3)O2 were prepared as active materials in electrochemical half-cells and were cycled electrochemically to obtain different values of Li concentration, x. Absorption edges of Ni, Mn, Co, and O in these materials of differing x were measured by electron energy loss spectrometry (EELS) in a transmission electron microscope to determine the changes in local electronic structure caused by delithiation. The work was supported by electronic structure calculations with the VASP pseudopotential package, the full-potential linear augmented plane wave code WIEN2K, and atomic multiplet calculations that took account of the electronic effects from local octahedral symmetry. A valence change from Ni2+ to Ni4+ with delithiation would have caused a 3 eV shift in energy of the intense white line at the Ni L3 edge, but the measured shift was less than 1.2 eV. The intensities of the "white lines" at the Ni L-edges did not change enough to account for a substantial change of Ni valence. No changes were detectable at the Mn and Co L-edges after delithiation either. Both EELS and the computational efforts showed that most of the charge compensation for Li+ takes place at hybridized O 2p states, not at Ni atoms. PMID- 16375322 TI - Characterization and transport properties of Nafion/polyaniline composite membranes. AB - Nafion membranes were modified by chemical polymerization of aniline using ammonium peroxodisulfate as the oxidant. The Nafion-polyaniline composite membranes were extensively characterized using scanning electron microscopy (SEM), atomic force microscopy (AFM), infrared (FTIR-ATR) and X-ray photoelectron spectroscopy (XPS), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and ion-exchange capacity measurements. The transport properties were also evaluated by conductivity and electrodialysis measurements. The data show that when a high oxidant concentration (1 M (NH4)2S2O8) is used, polyaniline is mostly formed at the surface of the Nafion membrane with a higher proportion of oligomers. On the contrary, when 0.1 M oxidant is used, polyaniline is mostly formed inside the ionic domains of Nafion, blocking the pathway to ion transport and thus reducing the transport of Zn2+ as well as the transport of H+. These data were also compared to the data obtained with poly(styrene sulfonate) PANI composite membranes. PMID- 16375323 TI - Comparison of UV and visible Raman spectroscopy of bulk metal molybdate and metal vanadate catalysts. AB - The visible (532 and 442 nm) and UV (325 nm) Raman spectra of bulk mixed metal oxides (metal molybdates and metal vanadates) were compared on the same spectrometer, for the first time, to allow examination of how varying the excitation energy from visible to UV affects the resulting Raman spectra. The quality of the Raman spectra was found to be a strong function of the absorption properties of the bulk mixed oxide. For bulk mixed metal oxides that absorb weakly in the visible and UV regions, both the visible and UV Raman spectra were of high quality and exhibit identical vibrational bands, but with slightly different relative intensities. For bulk mixed metal oxides that absorb strongly in the UV and visible regions and/or strongly in the UV and weakly in the visible regions, the visible Raman spectra are much richer in structural information and of higher resolution than the corresponding UV Raman spectra. This is a consequence of the strong UV absorption that significantly reduces the sampling volume and number of scatterers giving rise to the Raman signal. The shallower escape depth of UV Raman, however, was not sufficient to detect vibrations from the surface metal oxide species that are present on the outermost surface layer of these crystalline mixed metal oxide phases as previously suggested. It was also demonstrated that there is no sample damage by the more energetic UV excitation when very low laser powers and fast detectors are employed, thus avoiding the need of complicated fluidized bed sample arrangements sometimes used for UV Raman investigations. The current comparative Raman investigation carefully documents, for the first time, the advantages and disadvantages of applying different excitation energies in collecting Raman spectra of bulk mixed metal oxide materials. PMID- 16375324 TI - Iron-catalyzed propylene epoxidation by nitrous oxide: studies on the effects of alkali metal salts. AB - SBA-15-supported iron catalysts with and without alkali metal salt modifications were studied for propylene oxidation by nitrous oxide. The reaction route could be dramatically changed from allylic oxidation to epoxidation by modification of the FeOx/SBA-15 catalyst with alkali metal salts. The KCl-1 wt % FeOx/SBA-15 (K/Fe = 5) catalyst exhibited the best catalytic performances for propylene epoxidation, over which ca. 50% propylene oxide selectivity could be gained at a 10% propylene conversion at 648 K. Characterizations with diffuse reflectance UV Vis, XANES, and Raman spectroscopic techniques revealed that the modification with KCl increased the dispersion of the iron species and changed the local coordination of iron into a tetrahedral configuration on the inner surface of SBA 15. This tetrahedrally coordinated iron site, which was probably stabilized by potassium ions, was proposed to account for the epoxidation of propylene by nitrous oxide. At the same time, the reactivity of lattice oxygen was inhibited, and the acidity of the FeOx/SBA-15 was eliminated. These changes should also contribute to the increase in the selectivity to propylene oxide. The counteranions in the alkali metal salts exerted a significant influence on the catalytic behaviors probably via an electronic effect. PMID- 16375325 TI - Potential oscillations in galvanostatic electrooxidation of formic acid on platinum: a time-resolved surface-enhanced infrared study. AB - The mechanism of temporal potential oscillations that occur during galvanostatic formic acid oxidation on a Pt electrode has been investigated by time-resolved surface-enhanced infrared absorption spectroscopy (SEIRAS). Carbon monoxide (CO) and formate were found to adsorb on the surface and change their coverages synchronously with the temporal potential oscillations. Isotopic solution exchange (from H13COOH to H12COOH) and potential step experiments revealed that the oxidation of formic acid proceeds dominantly through adsorbed formate and the decomposition of formate to CO2 is the rate-determining step of the reaction. Adsorbed CO blocks the adsorption of formate and also suppresses the decomposition of formate to CO2, which raises the potential to maintain the applied current. The oxidative removal of CO at a high limiting potential increases the coverage of formate and accelerates the decomposition of formate, resulting in a potential drop and leading to the formation of CO. This cycle repeats itself to give the sustained temporal potential oscillations. The oscillatory dynamics can be explained by using a nonlinear rate equation originally proposed to explain the decomposition of formate and acetate on transition metal surfaces in UHV. PMID- 16375326 TI - Preparation and characterization of silica supported Au-Pd model catalysts. AB - Au-Pd bimetallic model catalysts were synthesized as alloy clusters on SiO2 ultrathin films under ultrahigh vacuum (UHV) conditions. The surface composition and morphology were characterized with low energy ion scattering spectroscopy (LEIS), infrared reflection absorption spectroscopy (IRAS), and temperature programmed desorption (TPD). Relative to the bulk, the surface of the clusters is enriched in Au. With CO as a probe, IRAS and TPD were used to identify isolated Pd sites at the surface of the supported Au-Pd clusters. Ethylene adsorption and dehydrogenation show a clear structure-reactivity correlation with respect to the structure/composition of these Au-Pd model catalysts. PMID- 16375327 TI - Vanadium oxides on aluminum oxide supports. 1. Surface termination and reducibility of vanadia films on alpha-Al2O3(0001). AB - Using density functional theory and statistical thermodynamics, we obtained the phase diagram of thin VnOm films of varying thickness (approximately 2-6 A, 1-6 vanadium layers) supported on alpha-Al2O3(0001). Depending on the temperature, oxygen pressure, and vanadium concentration, films with different thickness and termination may form. In ultrahigh vacuum (UHV), at room temperature and for low vanadium concentrations, an ultrathin (1 x 1) O=V-terminated film is most stable. As more vanadium is supplied, the thickest possible films form. Their structures and terminations correspond to previous findings for the (0001) surface of bulk V2O3 [Kresse et al., Surf. Sci. 2004, 555, 118]. The presence of surface vanadyl (O=V) groups is a prevalent feature. They are stable up to at least 800 K in UHV. Vanadyl oxygen atoms induce a V(2p) core-level shift of about 2 eV on the surface V atoms. The reducibility of the supported films is characterized by the energy of oxygen defect formation. For the stable structures, the results vary between 4.11 and 3.59 eV per 1/2O2. In contrast, oxygen removal from the V2O5(001) surface is much easier (1.93 eV). This provides a possible explanation for the lower catalytic activity of vanadium oxides supported on alumina compared to that of crystalline vanadia particles. PMID- 16375328 TI - Vanadium oxides on aluminum oxide supports. 2. Structure, vibrational properties, and reducibility of V2O5 clusters on alpha-Al2O3(0001). AB - The structure, stability, and vibrational properties of isolated V2O5 clusters on the Al2O3(0001) surface have been studied by density functional theory and statistical thermodynamics. The most stable structure does not possess vanadyl oxygen atoms. The positions of the oxygen atoms are in registry with those of the alumina support, and both vanadium atoms occupy octahedral sites. Another structure with one vanadyl oxygen atom is only 0.12 eV less stable. Infrared spectra are calculated for the two structures. The highest frequency at 922 cm( 1) belongs to a V-O stretch in the V-O-Al interface bonds, which supports the assignment of such a mode to the band observed around 941 cm(-1) for vanadia particles on alumina. Removal of a bridging oxygen atom from the most stable cluster at the V-O-Al interface bond costs 2.79 eV. Removal of a (vanadyl) oxygen atom from a thin vanadia film on alpha-Al2O3 costs 1.3 eV more, but removal from a V2O5(001) single-crystal surface costs 0.9 eV less. Similar to the V2O5(001) surface, the facile reduction is due to substantial structure relaxations that involve formation of an additional V-O-V bond and yield a pair of V(IV)(d1) sites instead of a V(III)(d2)/V(V)(d0) pair. PMID- 16375329 TI - Nanofaceted platinum surfaces: a new model system for nanoparticle catalysts. AB - We present a novel model system for nanoparticle electrocatalysts. A surface consisting of alternating (100) and (111) facets, several nanometers across and nearly 1 microm long, were self-assembled by annealing Pt single crystal surfaces initially cut at the midpoint between [111] and [100] directions, i.e., Pt(1+ square root of 3 1 1). The formation of these self-assembled arrays of nanofacets was monitored by in-situ surface X-ray scattering. These surfaces were further characterized with scanning probe microscopy and cyclic voltammetry. We found that the Pt(1+ square root of 3 1 1) surface is flat with less than 1 nm rms roughness when it was annealed in argon/hydrogen atmosphere. Then the surface forms nanofacets when it is annealed in pure air. This nanofaceting transition was completely reversible and reproducible. We investigated effects of CO adsorption on the voltammetric characteristics of both hydrogen-annealed and air annealed surfaces. We found that CO-adsorption/desorption cycles in CO containing electrolyte solution result in considerable modification of blank cyclic voltammograms for the both surfaces. We attributed these differences to the electrochemical annealing of surface defects due to the increased mobility during the cycles. PMID- 16375330 TI - Investigation of oxygen reduction reaction kinetics at (111)-(100) nanofaceted platinum surfaces in acidic media. AB - The oxygen reduction reaction (ORR) was studied on CO-treated and untreated (111) (100) nanofaceted platinum surfaces [Komanicky et al. J. Phys. Chem. 2005, 109, 23543] in sulfuric and perchloric acids using the rotating disk electrode technique. Activities of nanofaceted surfaces are found to be considerably higher than a simple average of the activities of (111) and (100) surfaces. We find that the high activity in sulfuric acid is consistent with the higher activity of (111) facets. It is due the weaker sulfate adsorption on finite-size (111) surfaces than on (111) single crystal surfaces where the ORR activity is suppressed by strong sulfate adsorption. However, the high activity found in the weakly absorbing perchloric acid cannot be explained by the finite-size effect, since the activities are reportedly insensitive to terrace sizes [Macia, M. D.; et al. J.Electroanal. Chem., 2004, 564, 141]. We propose a cooperative activity, unique to nanoscale objects, which results from oxy species crossing over between adjacent facets in nanometer proximities. PMID- 16375331 TI - Energy level alignment at metal-octaethylporphyrin interfaces. AB - We studied the electronic structure of copper-octaethylporphyrin (CuEOP) adsorbed on three metal surfaces--Ag(001), Ag(111), and Cu(111)--by means of ultraviolet photoelectron spectroscopy (UPS). The adsorption-induced work function shifts saturate roughly beyond two monolayers. The saturation values are substrate dependent, negative, and range from -1.30 to -0.85 eV. This shift is larger than that for tetraphenylporphyrins. The two highest occupied molecular orbitals (HOMO and HOMO-1) of the organic are clearly resolved in the UPS spectra. The origin of the negative work function shift is discussed. PMID- 16375332 TI - Nature of the Lewis acid sites on molybdenum and ruthenium sulfides: an electrostatic potential study. AB - The energy of formation and the Lewis acid strength of sulfur vacancies or coordinative unsaturated sites on the MoS2 edges were studied using density functional theory for periodic systems and an electrostatic potential-based methodology. The results suggest that the more energetically favorable sites are located on the sulfur edges; however, their Lewis acid strength is considerably smaller than the site acidity at the molybdenum edges. The acid strength for the reported most hydrodesulfurization active site of RuS2 was also determined. In general, the Lewis acid for the site on RuS2 is 100% smaller than the sites on the Mo edges and around 20% larger than the most favorable site on the S edges of MoS2. Binding of the pyridine molecule in the eta1 adsorption configuration on the considered sites has corroborated the trend of Lewis acidity suggested by the electrostatic potential methodology. PMID- 16375333 TI - Density functional theory study of water activation and COads + OHads reaction on pure platinum and bimetallic platinum/ruthenium nanoclusters. AB - A density functional theory study of the elementary steps that lead to the removal of CO(ads(Pt)) over alloyed and sequentially deposited Pt/Ru bimetallic nanoclusters is presented. The reaction energies and activation barriers for the H2O(ads(Ru)) dissociation and CO(ads(Pt)) + OH(ads(Ru)) reaction are estimated in solid-gas interface and in a microsolvated environment to determine which surface morphology is more tolerant to COads poisoning. On the basis of the energetics, the sequentially deposited Pt/Ru nanocluster is predicted to be a much more promising anode catalyst than the alloy cluster surface in fuel cell applications. PMID- 16375334 TI - MD simulations of the binding of alcohols and diols by a calixarene in water: connections between microscopic and macroscopic properties. AB - We report results of molecular dynamics (MD) simulations of the complexes of p sulfonatocalix[4]arene with linear alcohols from ethanol to heptanol in water at 25 degrees C. We show that these complexes are of the inclusion type and are governed by van der Waals interactions between the calixarene cavity and the inserted alkyl chain of the alcohol. We establish a correlation between the experimental Delta(r)H degrees values and the number of atoms inserted into the calixarene cavity. We also focus on the desolvation of the host and guest to establish the importance, at the enthalpic level, of the formation of hydrogen bond bridges between the calixarene and the alcohol molecule. The fact that methanol is not complexed by p-sulfonatocalix[4]arene is explained by calculating the cost of the desolvation of the guest upon complexation. We complete this study by modeling the complexes formed with 1,4-butanediol and 1,5-pentanediol. To explain the difference between the thermodynamic properties for the binding of 1,4-butanediol and butanol, we examine the insertion rate and the solvation of each hydroxy group. We show a specific behavior of one of the two hydroxy groups at the structural and energetic levels. PMID- 16375335 TI - Monte Carlo simulation of equilibrium reactions at vapor-liquid interfaces. AB - Chemical reactions are known to behave differently, depending upon their local environment. While the interactions with neighboring molecules may alter both the kinetics of chemical reactions and the overall equilibrium conversion, we have performed simulations of the latter. The particular environment that we address is the vapor-liquid interface, since only a few, limited studies have explored the influence of an interface on equilibrium reaction behavior. Simple dimerization reactions are modeled, as well as more complex multicomponent reactions, using the reactive Monte Carlo (RxMC) simulation technique. We find that the conversion of a reaction can be markedly different at an interface as compared to the bulk vapor and liquid phases, and these trends are analyzed with respect to specific intermolecular interactions. In conjunction, we calculate the surface tension of the reacting fluids at the interface, which is found to have unusual scaling behavior, with respect to the system temperature. PMID- 16375336 TI - Solubility of methane in water: the significance of the methane-water interaction potential. AB - The influence of the methane-water interaction potential on the value of the Henry constant obtained from molecular dynamics simulations was investigated. The SPC, SPC/E, MSPC/E, and TIP3P potentials were used to describe water and the OPLS UA and TraPPE potentials for methane. Nonbonding interactions between unlike atoms were calculated both with one of four mixing rules and with our new methane water interaction potential. The Henry constants obtained from simulations using any of the mixing rules differed significantly from the experimental ones. Good agreement between simulation and experiment was achieved with the new potential over the whole temperature range. PMID- 16375337 TI - Hydroxyl radical and hydroxide ion in liquid water: a comparative electron density functional theory study. AB - Ab initio density functional theory molecular dynamics simulations of the solvated states of the hydroxyl radical and hydroxide ion are performed using the Becke-Lee-Yang-Parr (BLYP) exchange-correlation functional (Becke, A. D. Phys. Rev. A 1988, 38, 3098. Lee, C.; Yang, W.; Parr, R. G. Phys. Rev. B 1988, 37, 785). The structures of the solvation shells of the two species are examined. It is found that the OH radical forms a relatively well-defined solvation complex with four neighboring water molecules. Three of these molecules are hydrogen bonded to the OH, while the fourth is hemibonded via a three-electron two centered bond between the oxygen atoms of the OH and water. The activity and the diffusion mechanism of the OH radical in water is discussed in comparison with the OH- ion. Although the results are partially influenced by the tendency of the BLYP density functional to overestimate hemibonded structure, the present simulations suggest that the widely accepted picture of rapid diffusion of OH radical in water through hydrogen exchange reaction may need to be reconsidered. PMID- 16375338 TI - Water's hydrogen bonds in the hydrophobic effect: a simple model. AB - We propose a simple analytical model to account for water's hydrogen bonds in the hydrophobic effect. It is based on computing a mean-field partition function for a water molecule in the first solvation shell around a solute molecule. The model treats the orientational restrictions from hydrogen bonding, and utilizes quantities that can be obtained from bulk water simulations. We illustrate the principles in a 2-dimensional Mercedes-Benz-like model. Our model gives good predictions for the heat capacity of hydrophobic solvation, reproduces the solvation energies and entropies at different temperatures with only one fitting parameter, and accounts for the solute size dependence of the hydrophobic effect. Our model supports the view that water's hydrogen bonding propensity determines the temperature dependence of the hydrophobic effect. It explains the puzzling experimental observation that dissolving a nonpolar solute in hot water has positive entropy. PMID- 16375339 TI - Relative Gibbs energies in solution through continuum models: effect of the loss of translational degrees of freedom in bimolecular reactions on Gibbs energy barriers. AB - We present here a cell model for evaluating Gibbs energy barriers corresponding to bimolecular reactions (or processes of larger molecularity) in which a loss of translational degrees of freedom takes place along the reaction coordinate. With this model, we have studied the Walden inversion processes: Xa- + H3CXb --> XaCH3 + Xb- (X = F, Cl, Br, and I). In these processes, our model yields an increase of about 2.3-3.4 kcal/mol in Gibbs energy in solution corresponding to the loss of the translational degrees of freedom when passing from separate reactants to the TS in good agreement with experimental data. The corresponding value in the gas phase is about 6.7-7.1 kcal/mol. When the difference between these two figures is used to correct the results obtained by the standard UAHF implementation of the continuum model, the theoretical results are brought significantly closer to the experimental ones. This seems to indicate that for these reactions the parametrization used does not adequately introduce the increase in Gibbs energy corresponding to the constriction of the translational motion of the species along the reaction coordinate when passing from the gas phase to solution. Therefore, we believe that continuum models could perform much better if we released the parametrization process from the task of taking into account the constriction in translation motion in solution, which could be more adequately evaluated using the cell model proposed here, thus allowing it to focus on better reproducing all the remaining solvation effects. PMID- 16375340 TI - Nonequilibrium potential function of chemically driven single macromolecules via Jarzynski-type Log-Mean-Exponential Heat. AB - Applying the method from recently developed fluctuation theorems to the stochastic dynamics of single macromolecules in ambient fluid at constant temperature, we establish two Jarzynski-type equalities: (1) between the log-mean exponential (LME) of the irreversible heat dissiption of a driven molecule in nonequilibrium steady-state (NESS) and ln P(ness)(x) and (2) between the LME of the work done by the internal force of the molecule and nonequilibrium chemical potential function mu(ness)(x) identical with U(x) + k(B)T ln P(ness)(x), where P(ness)(x) is the NESS probability density in the phase space of the macromolecule and U(x) is its internal potential function. Psi = integral mu(ness)(x) P(ness)(x) dx is shown to be a nonequilibrium generalization of the Helmholtz free energy and DeltaPsi = DeltaU - TDeltaS for nonequilibrium processes, where S = - kB integralP(x) ln P(x) dx is the Gibbs entropy associated with P(x). LME of heat dissipation generalizes the concept of entropy, and the equalities define thermodynamic potential functions for open systems far from equilibrium. PMID- 16375341 TI - Ion pair formation in water. association constants of bolaform, bisquaternary ammonium, electrolytes by chemical trapping. AB - The first and second association constants, K1 and K2, for ion pair formation in aqueous 0.02-3.5 M solutions of bis(trimethyl)-alpha,omega-alkanediammonium halides with variable spacer lengths, 1-n-1 2X (n = 2-4, X = Cl, Br) and bolaform salts and for tetramethylammonium halides (TMAX, X = Cl, Br), K(TMAX), were determined by the chemical trapping method. Values for K(TMAX) are small, K(TMABr) = 0.83 M(-1) and K(TMACl) = 0.29 M(-1), in agreement with literature values. For the bolaform salts, K1 depends on spacer length and counterion type, ranges from 0.4 to 17 M(-1), is 2-10 times larger than K2, is larger for Br- than Cl-, and decreases by a factor of approximately 3 for Cl- and approximately 10 for Br- as n increases from 2 to 4. K2, for the formation of bolaform dihalide pair, is essentially the same as that for ion pair formation in TMAX solutions, i.e., K2 approximately K(TMAX). Values of K1 and K(TMABr) obtained from changes in 79Br line widths are in good agreement with those obtained by chemical trapping. The results are consistent with a thermodynamic model in which the ion association depends on the balance of the ion specific hydration free energies of cations and anions and their ion specific and hydration interactions in ion pairs. Spacer length dependent ion pairing by bolaform electrolytes, which are analogues of the headgroups and counterions of gemini amphiphiles, suggests a new model for the spacer length dependent sphere-to-rod transitions of gemini micelles. Neutral, but polar, headgroup-counterion pairs have a lower demand for hydration that free headgroups and counterions, and headgroup-counterion pair formation releases interfacial water into the bulk aqueous phase, permitting tighter amphiphile packing in rodlike micelles. PMID- 16375342 TI - Hybrid ab initio VB/MM method--a valence bond ride through classical landscapes. AB - The development of a new hybrid (QM/MM) method, where the QM part is treated by ab initio valence bond (VB) theory is presented. This VB/MM method has the advantages of empirical VB (EVB) methodology but does not rely on empirical parameterization for the quantum part. The method implements embedding of the quantum region of each diabatic state separately, by treating the electrostatic interactions between QM and MM regions classically. Additionally, it assumes that changes of the off diagonal matrix element due to different environments are such that the overall resonance integral does not change. These assumptions are discussed in detail and the validity of the method is shown to be successful in three different bond dissociation processes, where bond dissociation as well as solvation energies compare very well with the experimental data. PMID- 16375343 TI - Transition state ensemble for the folding of B domain of protein A: a comparison of distributed molecular dynamics simulations with experiments. AB - Folding pathways of the B domain of staphylococcal protein A have been sampled with a distributed computing approach. Starting from an extended structure, the method employs an index measuring topological similarity to the native structure to selectively sample trajectory branches leading to the native fold. Unperturbed and continuous folding trajectories are drawn on a physics-based atomic potential energy surface with an implicit solvent. The sampled folding trajectories demonstrate a similar sequence of events: the earlier stage involves a partial formation of helix 2 and to a less extent of helix 1 at their N terminals, followed by the hydrophobic collapse between residues F14, I17, and L18 on helix 1 and residues R28, F31, and I32 on helix 2, which results in the rigidification of the helix turn from R28 to I32. Helix 2 is then able to extend, allowing for the formation to turn 2. The above description explains one experimental result why a G30A mutant of the protein was observed to be the fastest folder among proteins of its size. And the ensemble of structures right before the final collapse is in good agreement with the transition state ensemble mapped by another recent experiment with Fersht Phi values. We emphasize that because the approach here does not provide quantifications of the free energy landscape, our model of the transition state ensemble emerges from comparisons of simulations and previous experimental results rather from the simulation results alone. On the other hand, as our approach does not rely on a low-dimensional free energy surface, it can complement methods based on the construction of free energy surfaces. PMID- 16375344 TI - Interaction of Pd and PdCl2 with cellulose: a theoretical investigation. AB - The pyrolytic fragmentation of cellulose in the presence of atomic palladium (Pd) and palladium(II) chloride (PdCl2) has been studied with use of hybrid density functional theory and cellobiose as a model for cellulose. The configuration changes in the host, rearrangement of geometries of the products, and the respective reaction energetics for different fragmentation pathways are analyzed. While Pd is found to undergo insertion at the beta-1,4-linkage oxygen (O1)-carbon (C-1) of the rings, Pd(II) chloride is observed to promote the cleavage of the chain as well as rearrangement of the rings. A detailed mechanism for the formation of levoglucosan from one of the fragments following the interaction with PdCl2 is also highlighted. PMID- 16375345 TI - DNA psi-condensation and reentrant decondensation: effect of the PEG degree of polymerization. AB - Psi-condensation of DNA fragments of about 4 kbp was induced by poly(ethylene glycol) (PEG), with degrees of polymerization ranging from 45 to 182, and univalent salt (NaCl). Using circular dichroism spectroscopy, we were able to accurately determine the critical amount of PEG needed to induce condensation, as a function of the NaCl concentration. A significant dependence on the PEG degree of polymerization was found. Phase boundaries determined for the multimolecular condensation were very similar to those observed previously for the monomolecular collapse, with two asymptotic regimes at low and high salt concentrations. We analyze our data using a theoretical model that properly takes into account both the polyelectrolyte nature of the DNA and the liquid crystallinity of the condensed phase. The model assumes that all PEG is excluded from the condensates and shows reentrant decondensation only at low salt. We also systematically study reentrant decondensation and find a very strong dependence on PEG molecular weight. At low PEG molecular weight, decondensation occurs at relatively low concentrations of PEG, and over a wide range of salt concentrations. This suggests that in the reentrant decondensation the flexible polymers used are not completely excluded from the condensed phase. PMID- 16375346 TI - Structural changes during the photocycle of photoactive yellow protein monitored by ultraviolet resonance raman spectra of tyrosine and tryptophan. AB - Photoactive yellow protein (PYP) is a bacterial blue light photoreceptor, and photoexcitation of dark-state PYP (PYP(dark)) triggers a photocycle that involves several intermediate states. We report the ultraviolet resonance Raman spectra of PYP with 225-250 nm excitations and investigate protein structural changes accompanying the formation of the putative signaling state denoted PYP(M). The PYP(M)-PYP(dark) difference spectra show several features of tyrosine and tryptophan, indicating environmental changes for these amino acid residues. The tyrosine difference signals show small upshifts with intensity changes in Y8a and Y9a bands. Although there are five tyrosine residues in PYP, Tyr42 and Tyr118 are suggested to be responsible for the difference signals on the basis of a global fitting analysis of the difference spectra at different excitation wavelengths and the crystal structure of PYP(dark). A further experiment on the Thr50-->Val mutant supports environmental changes in Tyr42. The observed upshift of the Y8a band suggests a weaker or broken hydrogen bond between Tyr42 and the chromophore in PYP(M). In addition, a reorientation of the OH group in Tyr42 is suggested from the upshift of the Y9a band. For tryptophan, the Raman bands of W3, W16, and W18 modes diminish in intensity upon formation of PYP(M). The loss of intensities is attributable to an exposure of tryptophan in PYP(M). PYP contains only one tryptophan (Trp119) that is located more than 10 A from the active site. Thus the observed changes are indicative of global conformational changes in protein during the transition from PYP(dark) to PYP(M). These results are in line with the currently proposed photocycle mechanism of PYP. PMID- 16375347 TI - Detecting protein-protein interactions by isotope-edited infrared spectroscopy: a numerical approach. AB - We present a theoretical and numerical analysis of the vibrational coupling between isotope-edited amino acids in protein dimers. Depending on the presence and magnitude of coupling between 13Calpha=O peptide bond oscillators, characteristic level splittings of vibrational eigenstates are predicted. For the example of the Gramicidin A ion channel polypeptide, we observe typical IR fingerprints for the head-to-head and the antiparallel double-helical conformation of the dimer. We suggest that these findings can be used to clearly identify the structure of polypeptide aggregates using a particularly simple isotope substitution pattern. PMID- 16375348 TI - Electron-transfer dynamics of photosynthetic reaction centers in thermoresponsive soft materials. AB - Poly(ethylene glycol)-grafted, lipid-based, thermoresponsive, soft nanostructures are shown to serve as scaffolding into which reconstituted integral membrane proteins, such as the bacterial photosynthetic reaction centers (RCs) can be stabilized, and their packing arrangement, and hence photophysical properties, can be controlled. The self-assembled nanostructures exist in two distinct states: a liquid-crystalline gel phase at temperatures above 21 degrees C and a non-birefringent, reduced viscosity state at lower temperatures. Characterization of the effect of protein introduction on the mesoscopic structure of the materials by 31P NMR and small-angle X-ray scattering shows that the expanded lamellar structure of the protein-free material is retained. At reduced temperatures, however, the aggregate structure is found to convert from a two dimensional normal hexagonal structure to a three-dimensional cubic phase upon introduction of the RCs. Structural and functional characteristics of the RCs were determined by ground-state and femtosecond transient absorption spectroscopy. Time-resolved results indicate that the kinetics of primary electron transfer for the RCs in the low-viscosity cold phase of the self assembled nanostructures are identical to those observed in a detergent solubilized state in buffered aqueous solutions (approximately 4 ps) over a wide range of protein concentrations and experimental conditions. This is also true for RCs held within the lamellar gel phase at low protein concentrations and at short sample storage times. In contrast are kinetics from samples that are prepared with high RC concentrations and stored for several hours, which display additional kinetic components with extended electron-transfer times (approximately 10-12 ps). This observation is tentatively attributed to energy transfer between RCs that have laterally (in-plane) organized within the lipid bilayers of the lamellar gel phase prior to charge separation. These results not only demonstrate the use of soft nanostructures as a matrix in which to stabilize and organize membrane proteins but also suggest the possibility of using them to control the interactions between proteins and thus to tune their collective optical/electronic properties. PMID- 16375349 TI - Vibrational spectral simulation for peptides of mixed secondary structure: method comparisons with the Trpzip model hairpin. AB - Infrared absorption and vibrational circular dichroism (IR and VCD) spectra of model fragments of TrpZip-style beta-hairpin structures are simulated using density functional theory (DFT) methods to estimate the influence of fragment size, end effects, conformational irregularities, peptide side chains, and solvent. Different fragmentation schemes, computing the strands and turn segments separately, were tested by varying the sizes of each and their respective overlaps. For suitably overlapping fragments, atomic property tensors were found to be reliably transferable, as tested by their ability to generate simulated spectra in good agreement with results from ab initio DFT computations for the entire peptide. This fragment approach significantly reduces computational times and opens up a wider range of systems that can be studied with a DFT-based approach as compared to previous methods based on uniform repeating sequences. However, vacuum calculations do not adequately represent the frequency dispersion of solvated molecules, and thus, some alternate strategies for solvation correction are explored for improving the simulation accuracy. Unlike for regular periodic secondary structure, the solvent significantly impacts the spectral shapes of hairpins, due to the different degrees of hydration of individual amide groups, which can be exposed to or shielded from water due to external vs internal hydrogen bonding. This is amplified by the shielding of selected amides from the solvent due to bulky side chains. The peptide plus solvent was structurally modeled with molecular dynamics methods, and then an electrostatic field-based parametrization correction was added to the force field and intensity tensors to compensate for the solvent dipolar field. The effect of the shielding and subsequent reordering of modes has a larger impact on VCD than IR band shapes. PMID- 16375350 TI - Structural transitions in polyriboadenylic acid induced by the changes in pH and temperature: vibrational circular dichroism study in solution and film states. AB - A correlation of the changes in vibrational absorption and vibrational circular dichroism (VCD) spectral features with the structural changes of polyriboadenylic acid (polyA) as a function of pH is reported. Analysis of the solution spectral data as a function of pH led us to establishing the importance of a previously unrecognized absorption band at approximately 1665 cm(-1). The present studies indicate that this absorption band and associated VCD originate from the double helical structure of polyA. The observed changes in solution-state VCD features are indicative of the pH-dependent transitions among the three acidic forms of polyA (A, B, and "frozen" form). In addition to the solution-state spectral data, pH-dependent absorption and VCD spectra for films of polyA, derived from dilute H2O solutions, are also presented. The pH-dependent changes in the absorption and VCD spectra of films are also correlated to the polyA structural changes. PMID- 16375351 TI - Second half-reaction of nitric oxide synthase: computational insights into the initial step and key proposed intermediate. AB - Density functional theory methods have been employed to investigate possible first steps in the second half-reaction of the mechanism of nitric oxide synthases (NOSs). In particular, reactions and complexes formed via transfer of either or both hydrogens of the substrates (NHA) -NHOH group to the Fe-bound O2 were considered. For each of these pathways, the effect of adding an extra electron from tetrahydrobiotperin (H4B) was also examined. The preferred initial pathway involves the simultaneous transfer of both hydrogens of the -NHOH group to the Fe(heme)-O2, without an additional electron, to give the Fe(heme)-HOOH species which lies only marginally higher in energy, 2.5 kcal mol(-1) or less, than the initial bound active site. An alternative mechanism in which only the NH- proton of the -NHOH group is transferred to the Fe(heme)-O2 to give an Fe(heme)-OOH derivative is found to require only slightly more energy, approximately 2 kcal mol(-1). However, transfer of the proton back to the -NOH nitrogen occurs without a barrier at 298.15 K. Tetrahedral intermediates in which the Fe(heme)-O2 has attached at the guanidinium carbon (C(guan)) of NHA, that is, forms an Fe(heme)-O2-C(guan) link, have also been investigated. All examples of such species considered, that is, with or without hydrogen or electron transfers, lie significantly higher in energy by at least 29.0 kcal mol(-1) than the initial bound active site. Thus, it is suggested that such complexes are not mechanistically feasible. The implications of the present findings for the second half-reaction are also discussed. PMID- 16375352 TI - Fluorescence resonance energy transfer in CdSe/ZnS-DNA conjugates: probing hybridization and DNA cleavage. AB - Nucleic-acid-functionalized CdSe/ZnS quantum dots (QDs) were hybridized with the complementary Texas-Red-functionalized nucleic acid. The hybridization was monitored by following the fluorescence resonance energy transfer from the QDs to the dye units. Treatment of the QD/dye DNA duplex structure with DNase I resulted in the cleavage of the DNA and the recovery of the fluorescence properties of the CdSe/ZnS QDs. The luminescence properties of the QDs were, however, only partially recovered due to the nonspecific adsorption of the dye onto the QDs. Similarly, nucleic-acid-functionalized Au nanoparticles (Au NPs) were hybridized with the complementary Texas-Red-labeled nucleic acid. The hybridization was followed by the fluorescence quenching of the dye by the Au NPs. Treatment of the Au NP/dye DNA duplex with DNase I resulted in the cleavage of the DNA and the partial recovery of the dye fluorescence. The incomplete recovery of the dye fluorescence originated from the nonspecific binding of the dye units to the Au NPs. The nonspecific binding of the dye to the CdSe/ZnS QDs and the Au NPs is attributed to nonprotected surface vacancies in the two systems. PMID- 16375353 TI - Polyphenols deriving from chalcones: investigations of redox activities. AB - The redox properties of a series of hydroxychalcones (a group of polyphenols abundantly present in plants) were investigated by cyclic voltammetry. As for many polyphenols, their beneficial properties have been mainly related to their antioxidant activities, which in turn are directly associated to their redox behavior. Two types of radicals can be produced that are localized on either one of the two aromatic systems. Their thermodynamic and kinetic parameters were extracted and compared to the predictions of density functional theory calculations. When at least one OH is present on each ring, their behaviors are dominated by the conjugated system: phenolic ring A-double bond-ketone, which is the only one to be oxidized. However, the redox properties of this conjugated system are strongly influenced by the presence of ring B. When an OH is present on ring B, an important feature is the existence of strong hydrogen bonding that remains almost unmodified even when ring A is oxidized. It does not considerably change the thermodynamics of ring A but strongly increases the rigidity of the molecule that remains planar under the neutral, anionic, or radical forms. Oxidation potentials of the phenolates range between 0.1 and 0.2 V versus a saturated calomel electrode, which correspond to species that are very easy to oxidize and lead to the rapid formation of nonradical species, underlining the potential antioxidant properties of these molecules. PMID- 16375355 TI - Spectrophotometric study of fluorescence sensing and selective binding of biochemical substrates by 2,2'-bridged bis(beta-cyclodextrin) and its water soluble fullerene conjugate. AB - A bis(beta-cyclodextrin)-fullerene conjugate (3) linked at the secondary hydroxyl side was prepared in a good yield from its precursor N,N'-bis(2-(2 aminoethylamino)ethyl)malonamide-bridged bis(beta-cyclodextrin) (2). Spectrophotomeric studies on the conformation and the inclusion complexation behavior of 3 with a variety of organic and biochemical substrates by means of UV vis, FT-IR, NMR, fluorescence, and circular dichroism spectroscopy showed that the bis(beta-cyclodextrin)-fullerene conjugate displayed an intramolecular capsule-type conformation in aqueous solution. Because of the multiple binding of bis(beta-cyclodextrin) with substrates, 2 can act as an efficient fluorescence sensor for biochemical substrates, while its fullerene conjugate 3 displays a capability of cleaving DNA under visible-light irradiation. PMID- 16375354 TI - Dynamic mechanism of E2020 binding to acetylcholinesterase: a steered molecular dynamics simulation. AB - The unbinding process of E2020 ((R,S)-1-benzyl-4-[(5,6-dimethoxy-1-indanon)-2-yl] methylpiperidine) leaving from the long active site gorge of Torpedo californica acetylcholinesterase (TcAChE) was studied by using steered molecular dynamics (SMD) simulations on a nanosecond scale with different velocities, and unbinding force profiles were obtained. Different from the unbinding of other AChE inhibitors, such as Huperzine A that undergoes the greatest barrier located at the bottleneck of the gorge, the major resistance preventing E2020 from leaving the gorge is from the peripheral anionic site where E2020 interacts intensively with several aromatic residues (e.g., Tyr70, Tyr121, and Trp279) through its benzene ring and forms a strong direct hydrogen bond and a water bridge with Ser286 via its O24. These interactions cause the largest rupture force, approximately 550 pN. It was found that the rotatable bonds of the piperidine ring to the benzene ring and dimethoxyindanone facilitate E2020 to pass the bottleneck through continuous conformation change by rotating those bonds to avoid serious conflict with Tyr121 and Phe330. The aromatic residues lining the gorge wall are the major components contributing to hydrophobic interactions between E2020 and TcAChE. Remarkably, these aromatic residues, acting in three groups as "sender" and "receiver", compose a "conveyer belt" for E2020 entering and leaving the TcAChE gorge. PMID- 16375356 TI - Hydration of a synthetic clay with tetrahedral charges: a multidisciplinary experimental and numerical study. AB - The interaction of water with a synthetic saponite clay sample, with a layer charge of 1 per unit cell (0.165 C m(-2)), was investigated by following along water adsorption and desorption in the relative pressure range from 10(-6) to 0.99 (i) the adsorbed amount by gravimetric and near-infrared techniques, (ii) the basal distance and arrangement of water molecules in the interlayer by X-ray and neutron diffraction under controlled water pressure, and (iii) the molecular structure and interaction of adsorbed water molecules by near-infrared (NIR) and Raman spectroscopy under controlled water pressure. The results thus obtained were confronted with Grand Canonical Monte Carlo (GC/MC) simulations. Using such an approach, various well-distinct hydration ranges can be distinguished. In the two first ranges, at low water relative pressure, adsorption occurs on external surfaces only, with no swelling associated. The next range corresponds to the adsorption of water molecules around the interlayer cation without removing it from its position on top of the ditrigonal cavity of the tetrahedral layer and is associated with limited swelling. In the following range, the cation is displaced toward the mid-interlayer region. The interlamellar spacing thus reached, around 12.3 A, corresponds to what is classically referred to as a "one-layer hydrate," whereas no water layer is present in the interlayer region. The next hydration range corresponds to the filling of the interlayer at nearly constant spacing. This leads to the formation of a well-organized network of interlayer water molecules with significant interactions with the clay layer. The structure thus formed leads to a complete extinction of the d001 line in D2O neutron diffraction patterns that are correctly simulated by directly using the molecular configurations derived by GC/MC. The next range (0.50 < P/P0 < 0.80) corresponds to the final swelling of the structure to reach d spacing values of 15.2 A (usually referred to the "two-layer hydrate"). It is associated with the development of a network of liquidlike water molecules more structured than in bulk water. The final hydration range at high relative pressure mainly corresponds to the filling of pores between clay particles. PMID- 16375357 TI - Surfactant/nonionic copolymer interaction: a SLS, DLS, ITC, and NMR investigation. AB - The interactions between an oxyphenylethylene-oxyethylene nonionic diblock copolymer with the anionic surfactant sodium dodecyl sulfate (SDS) have been studied in dilute aqueous solutions by static and dynamic light scattering (SLS and DLS, respectively), isothermal titration calorimetry (ITC), and 13C and self diffusion nuclear magnetic resonance techniques. The studied copolymer, S20E67, where S denotes the hydrophobic styrene oxide unit and E the hydrophilic oxyethylene unit, forms micelles of 15.6 nm at 25 degrees C, whose core is formed by the styrene oxide chains surrounded by a water swollen polyoxyethylene corona. The S20E67/SDS system has been investigated at a copolymer concentration of 2.5 g dm(-3), for which the copolymer is fully micellized, and with varying surfactant concentration up to approximately 0.15 M. When SDS is added to the solution, two different types of complexes are observed at various surfactant concentrations. From SLS and DLS it can be seen that, at low SDS concentrations, a copolymer-rich surfactant mixed micelle or complex is formed after association of SDS molecules to block copolymer micelles. These interactions give rise to a strong decrease in both light scattering intensity and hydrodynamic radius of the mixed micelles, which has been ascribed to an effective reduction of the complex size, and also an effect arising from the increasing electrostatic repulsion of charged surfactant-copolymer micelles. At higher surfactant concentrations, the copolymer rich surfactant micelles progressively are destroyed to give surfactant-rich copolymer micelles, which would be formed by a surfactant micelle bound to one or very few copolymer unimers. ITC data seem to confirm the results of light scattering, showing the dehydration and rehydration processes accompanying the formation and subsequent destruction of the copolymer-rich surfactant mixed micelles. The extent of interaction between the copolymer and the surfactant is seen to involve as much as carbon 3 (C3) of the SDS molecule. Self-diffusion coefficients corroborated light scattering data. PMID- 16375358 TI - Modeling water exchange on an aluminum polyoxocation. AB - For the first time, water exchange on a polymeric complex has been modeled using a combination of gas-phase ab initio calculations and molecular dynamics (MD) simulations. The GaO4Al12(OH)24(H2O)12(7+)aq ion (GaAl12) was chosen because high quality experimental data exist, including an activation enthalpy (+63 +/- 7 kJ/mol) and an activation volume (+3 +/- 1 cm3/mol). We took a two-step approach. First, the local solvent structure and the initial states for reaction were inferred from the molecular dynamics simulations. Second, we used this information to evaluate initial-state structures in the ab initio calculations. The energy differences between the initial and transition states from the ab initio calculations varied from +59 kJ/mol to +53 kJ/mol depending upon details, closely approximating the activation enthalpy. PMID- 16375359 TI - Electron transport in coumarin-dye-sensitized nanocrystalline TiO2 electrodes. AB - We investigated electron transport kinetics in terms of electron diffusion coefficient (D) and electron lifetime (tau) in coumarin-dye-sensitized nanocrystalline TiO2 electrodes by intensity-modulated photocurrent spectroscopy (IMPS) and intensity-modulated photovoltage spectroscopy (IMVS). We found that the values of tau for coumarin-dye-sensitized TiO2 electrodes were much shorter than that for an electrode coated with a Ru complex (N719 dye), suggesting that the back-electron-transfer process corresponding to recombination between conduction-band electrons in the TiO2 and I3- ions in the electrolyte occurs more easily in coumarin-dye-sensitized solar cells. In addition, the values of tau depended on the kind of coumarin dye, each of which has a different number of thiophene moieties, suggesting that the molecular structure of the adsorbed dyes also affects the kinetics of electron transport in the TiO2 electrodes. PMID- 16375360 TI - In situ STM study of potential-dependent height change of a tetrathiafulvalene derivative embedded in alkanethiol self-assembled monolayers on Au(111). AB - Electroactive tetrathiafulvalene thiol islands embedded in an n-alkanethiol SAM matrix were studied under potential control using in situ scanning tunneling microscopy. Unlike previously studied stochastic switching, the apparent height of the island could be intentionally controlled in the present system by choosing the appropriate potential and the island size. The dependence of the height change on the potential and the size is explained as structural change of the island induced by the charging effect of the electroactive moiety. PMID- 16375361 TI - Ordered alignment of CdS nanocrystals on MWCNTs without surface modification. AB - This paper describes a facile hydrothermal procedure capable of aligning CdS nanocrystals on multiwalled carbon nanotubes (MWCNTs) without the use of organic bridging molecules such as cysteamine and carboxylates. The direct placement of CdS on MWCNTs allows good mixing and better interfacing between the two nanophases. The synthesis conditions can in principle be tuned to produce modulations in compositions, phases, and crystal orientations to the need of optoelectronic and photonic applications. PMID- 16375362 TI - Electrochemical growth of two-dimensional gold nanostructures on a thin polypyrrole film modified ITO electrode. AB - Two-dimensional gold nanostructures have been fabricated by electrochemical deposition of gold nanoparticles onto indium tin oxide (ITO) glass substrate modified with thin polypyrrole film. By controlling the electrodeposition conditions, gold nanoparticles with dendritic rod, sheet, flower-like (consisting of staggered nanosheets), and pinecone-like structures were generated. The flower like gold nanoparticles showed high catalytic activity on electrochemical reduction of oxygen, and its activity was measured to be approximately 25 times that of gold pinecones and 10(4) times that of gold nanosheets in terms of gold weight. The pinecone-like nanoparticles can form a compact film with nano /microscale binary structure like a lotus leaf surface. After modification with n dodecanethiol, the surface showed superhydrophobic properties with a water contact angle of 153.4 degrees and a tilt angle of 4.4 degrees (5 microL droplet). PMID- 16375363 TI - Influence of pore dimension and sorption configuration on the heat of sorption of hexane on monodimensional siliceous zeolites. AB - Sorption of n-hexane on monodimensional pure silica SSZ-35, CIT-5, ZSM-12, and ZSM-22 zeolites with different pore dimension and on recently synthesized ITQ-29 was studied by IR spectroscopic and computational chemistry methods. Heats of sorption of n-hexane on these zeolites was determined experimentally from the temperature dependence of the intensity of IR bands of sorbed hexane as well as from theoretical calculations. Calculations have shown the different orientations of sorbed hexane molecules inside zeolite channels, which depend on the type of zeolite and loading. At high loadings, ordering of hexane inside the channels is observed due to optimization of sorbate-sorbate and sorbate-zeolite interaction energies. Such ordering is responsible for the increase of the sorption energy. A decrease of the sorption energy upon increasing the pore dimension of zeolite was observed, in agreement with results previously published in the literature. Effects of pore diameter of zeolites and ordering of molecules inside zeolite channels on the sorption energy of hexane are discussed. PMID- 16375364 TI - V4S9Br4: a novel high-spin vanadium cluster thiobromide with square-planar metal core. AB - The novel vanadium thiobromide, V4S9Br4, with a square-planar metal cluster core was synthesized and characterized by single-crystal X-ray diffraction, measurements of magnetic properties and the heat capacity, and DFT calculations of the electronic structure. At the room temperature, the compound displays paramagnetic properties with an independent spin on each V atom and with a weak exchange constant (J approximately 10 cm(-1)). The paramagnetic state is transformed into a low-spin state (AF-type ordering) at low temperatures. This change is accompanied by a heat-capacity anomaly. The observed magnetic and heat capacity anomalies can be explained by the thermal excitation of electrons on the closely spaced molecular energy levels in the presence of the Jahn-Teller effect. PMID- 16375365 TI - Electromagnetic coupling in near-field scattering by small homogeneous and heterogeneous nanoaggregates. AB - Progress in near-field optical spectroscopy research on metal nanoparticles demands a better understanding of the role played by particle-particle interactions and a deeper insight of the influence of the incident field wavelength. This is particularly true for scanning near-field optical microscopy (SNOM), where the mechanism by which some components of the evanescent illuminating field are transformed into propagating field components that carry information about the sample is at the core of the image formation and where the role played by the interactions between sample and tip remains a still open problem. In this perspective, we investigate numerically the optical behavior of small aggregates of spherical nanoparticles, taking into account the electromagnetic coupling between all particles and the apertureless tip. The tip is modeled as a sphere made of different materials characterized by appropriate dielectric functions. We find that the tip material affects both qualitatively and quantitatively the SNOM images; more important, from the analysis of the calculated scattering cross section, the resonance plasmon location of the whole (aggregate + tip) system undergoes detectable changes, if the tip is constituted of the same material of the sample, as the tip is situated in different positions. This modification of the plasmon frequencies induces a nontrivial variation of the near-field intensity as a function of the tip position and the resulting SNOM image can be distorted with respect to the actual shape of the sample. No simple arguments can be used to relate the value of the local field on the tip surface to the scattering cross section value; depending on the tip material, the comparison between these two measurements can help to clarify the role of basic interactions in the scattering mechanism. PMID- 16375366 TI - High conversion synthesis of pyrene end functionalized polyrotaxane based on poly(ethylene oxide) and alpha-cyclodextrins. AB - We describe the quantitative synthesis of new pyrene labeled cyclodextrin-based polyrotaxane starting from pseudopolyrotaxane of alpha,omega-dimethacrylate poly(ethylene oxide) (PEO) and alpha-cyclodextrins (alpha-CDs). Using a solvent mixture (H2O/dimethyl sulfoxide (DMSO)), an almost quantitative conversion in polyrotaxane can be achieved using the coupling reaction between methacrylic functions and 1-pyrene butyric acid N-hydroxysuccinimide ester. This result is due to the fast blocking reaction of the pseudopolyrotaxane telechelic functions. The polyrotaxanes are characterized by NMR, size exclusion chromatography (SEC), and small-angle neutron scattering (SANS). A rodlike structure of the polyrotaxane is evidenced by SANS, and a persistence length of 70 A is determined. This result corresponds to an almost completely stretched PEO chain of 1000 g.mol(-1) molecular weight. We furthermore studied the opposite case of low packing density polyrotaxanes that were also silylated to suppress interactions between cyclodextrins. We observed a random coil structure only for silylated low packed polyrotaxane. This result demonstrates that both hydrogen bonding and packing density can explain the rodlike structure of cyclodextrin based polyrotaxane. PMID- 16375367 TI - Wall "thickness" effects on Raman spectrum shift, thermal conductivity, and Young's modulus of single-walled nanotubes. AB - We demonstrate that at a finite temperature, an effective wall thickness of a single-walled carbon nanotube (SWNT) should be W = Ws + Wd, where Ws is the static thickness defined as the extension of the outmost electronic orbit and Wd the dynamic thickness due to thermal vibration of atoms. Both molecular simulations and a theoretical analysis show that Wd is proportional to T1/2. We find that the increase of dynamic thickness with temperature is the main mechanism of Raman spectrum shift. The introduction of the dynamic thickness changes some conclusions about Young's modulus and reduces the values of thermal conductivity. PMID- 16375368 TI - Relaxation dynamics of 2,7- and 3,6-distyrylcarbazoles in solutions and in solid films: mechanism for efficient nonradiative deactivation in the 3,6-linked carbazole. AB - We performed time-resolved spectral investigations of two distyrylcarbazole derivatives, 2,7- and 3,6-distyrylcarbazole (2,7-DPVTCz and 3,6-DPVTCz, respectively), in dilute toluene solution and in solid films mixed with poly(methyl methacrylate) (PMMA). The lifetime of 2,7-DPVTCz in its excited state in solution is approximately 100 times as great as that of 3,6-DPVTCz, consistent with their photophysical nature. The former shows intense emission, but the latter is nearly nonfluorescent in a free environment. Moreover, the lifetime of 3,6-DPVTCz in its excited state increased also approximately 100 times when the molecule was encapsulated in a 3,6-DPVTCz/PMMA solid film, indicating that intramolecular motion of the molecule significantly affects the observed relaxation dynamics in a confined environment. Calculations on the excited states indicate that an efficient intersystem crossing is activated upon twisting of the bridged C-C single bond in a free 3,6-linked carbazole; such efficient deactivation is impractical in 2,7-linked carbazole or for 3,6-linked carbazole in a PMMA matrix. Information obtained from experiments on femtosecond fluorescence enables us to distinguish crucial relaxation processes in the excited state for a profound understanding of the details of vibrational and electronic relaxations of 3,6-DPVTCz in solution. PMID- 16375369 TI - Hydride ion as a two-electron donor in a nanoporous crystalline semiconductor 12CaO.7Al2O3. AB - The 12CaO.7Al2O3 (C12A7) crystal with a nanoporous lattice framework exhibits high electrical conductivity with an activation energy of approximately 1.5 eV when equilibrated in a hydrogen atmosphere above approximately 800 degrees C. The high conductivity is preserved in a quenched state below approximately 600 degrees C with a reduced activation energy of approximately 0.8 eV. Such complex behavior in electrical conductivity is associated with incorporation of hydride ions (H-) in cages of the lattice framework. Electromotive force measurements reveal that the major carrier for the conductivity is electron with a small contribution by proton (H+), ruling out the possibility of direct intercage migration of the H- ion. A combination of these observations with the ab initio calculations leads to the conclusion that the electrons are thermally generated from the H- ion by the dissociation into two electrons and an proton, which is further converted to an OH- ion via reaction with an extraframework oxide ion (O2 ). The energy difference between the initial (H- + O2-) and the final (2e- + OH-) states as evaluated by the theoretical calculation is as small as approximately 1 eV, which agrees well with an experimentally obtained enthalpy change, approximately 1.4 eV. Thus, internal equilibration between the extraframework hydrogen and the oxygen species is responsible for the thermal generation of the carrier electron. It is also suggested that the same conductive (2e- + OH-) state is reached by the photoirradiation of H- -containing C12A7. In this case the photoionization of H- forms an electron and an Ho atom, which then forms an OH- ion and another electron with thermal assistance. The persistence of photoinduced conductivity is explained by the slow kinetics of the reverse process at room temperature. PMID- 16375370 TI - Ion-exchange controls the kinetics of deswelling of polyelectrolyte microgels in solutions of oppositely charged surfactant. AB - The kinetics of deswelling of sodium polyacrylate microgels (radius 30-140 microm) in aqueous solutions of dodecyltrimethylammonium bromide is investigated by means of micropipet-assisted light microscopy. The purpose of the study is to test a recent model (J. Phys. Chem. B 2003, 107, 9203) proposing that the rate of the volume change is controlled by the transport of surfactant from the solution to the gel core (ion exchange) via the surfactant-rich surface phase appearing in the gel during the volume transition. Equilibrium swelling characteristics of the gel network in surfactant-free solutions and with various amounts of surfactant present are presented and discussed with reference to related systems. A relationship between gel volume and degree of surfactant binding is determined and used in theoretical predictions of the deswelling kinetics. Experimental data for single gel beads observed during deswelling under conditions of forced convection are presented and compared with model calculations. It is demonstrated that the dependences of the kinetics on initial gel size, the surfactant concentration in the solution, and the liquid flow rate are well accounted for by the model. It is concluded that the deswelling rates of the studied gels are strongly influenced by the mass transport of surfactant between gel and solution (stagnant layer diffusion), but only to a minor extent by the transport through the surface phase. The results indicate that, during the volume transition, swelling equilibrium (network relaxation/transport of water) is established on a relatively short time scale and, therefore, can be treated as independent of the ion-exchange kinetics. Theoretical aspects of the kinetics and mechanisms of surfactant transport through the surface phase are discussed. PMID- 16375371 TI - Small-angle X-ray scattering and light scattering on lysozyme and sodium glycocholate micelles. AB - Small-angle X-ray scattering (SAXS) together with static (SLS) and dynamic light scattering (DLS) measurements were carried out on aqueous solutions of lysozyme (LY) and of the ionic biological detergent sodium glycocholate (NaGC). Apparent diffusion coefficients (D app), excess Rayleigh ratio, and SAXS spectra were measured for 0.1 M NaGC solutions at different ionic strengths (0.05-0.30 M NaCl). The same data were collected for LY in sodium acetate buffer 50 mM without and with 92 mM NaCl as a function of protein concentration (10-80 g L(-1)). A correlated analysis of SLS data and SAXS spectra was first tested on the LY samples and then extended to the interpretation of the NaGC data to infer information on particle structure and interaction potential. A hard-core (HC) interaction shell of uniform thickness, a screened Coulomb potential of the electric double layer (EDL) or the complete DLVO potential were alternatively used to represent the long-range tail of the interaction potential. Whenever an essentially repulsive tail is expected, all the representations give reasonable results, but the data analysis does not allow the discrimination between the oblate and the prolate symmetries of the NaGC aggregates. The DLVO model allows the interpretation of the data even when the attractive component determines the tail character. With this model an overall fit of the micelle data at all the NaCl concentrations was successfully performed by assuming a simple spherical symmetry of the micelles and invariant values of their ionization degree and Hamaker constant, thus considering just the screening effect of the added electrolyte. Whatever model is used, the results point out that the aggregates are quite hydrated (26-38 water molecules per monomer) and very slightly grow by increasing the NaCl concentration. When spherical symmetry is assumed the aggregate radii for all the samples fall in the range 15-16 A. From the SAXS and SLS, best fitting geometrical parameters, and interparticle structure factor, a D app value was calculated for each sample. An excellent consistence is achieved for LY results. On the contrary, calculated D app values systematically lower than the experimental values are always obtained for the NaGC micelles. Micelle polydispersity and internal dynamics seem to be the most probable reasons of the bad agreement. PMID- 16375372 TI - Growth versus cyclization in the early stages of the polycondensation of metal alkoxides. AB - The early steps of the polycondensation of transition metal alkoxide have been studied from the chemical and structural points of view. Polyoxoalkoxides are described like macromolecules by the composition of the repeating unit, the degree of polymerization (N), and the radius of gyration (R). The fraction p of binding sites of the coordination sphere of the metal centers occupied by terminal ligands determines N as follows: N proportional, variant pdf (dA-df), where df is the fractal dimension and dA is defined by Np proportional, variant RdA. This approach addresses difficulties raised by both coordinative unsaturation and cyclization in the modelization of the polycondensation of metal alkoxides. The coordinative unsaturation is accounted for by a particularly small value of dA= 1 in the very early steps, while the cyclization frequency is measured by the difference dA-df. This difference is not constant along the polycondensation process, and its dependence on the extent of reaction provides clues for understanding the high apparent kinetics order of gelation often reported in the literature. PMID- 16375373 TI - A mathematical model for crystal growth by aggregation of precursor metastable nanoparticles. AB - A mathematical model is developed to describe aggregative crystal growth, including oriented aggregation, from evolving pre-existing primary nanoparticles with composition and structure that are different from that of the final crystalline aggregate. The basic assumptions of the model are based on the ideas introduced in an earlier published report [Buyanov and Krivoruchko, Kinet. Katal. 1976, 17, 666-675] to describe the growth of low-solubility metal hydroxides (e.g., iron oxides) by oriented aggregation. It is assumed that primary particles can be described as pseudo-species A, B, and C, which have the following properties: (1) fresh primary particles (colloidally stable inert nanoparticles, denoted as A), (2) mature primary particles (partially transformed nanoparticles at an optimum stage of development for attachment to a growing crystal, denoted as B), and (3) nucleated primary particles (denoted as C1). The evolution of primary particles, A --> B --> C1, is treated as two first-order consecutive reactions. Crystal growth via crystal-crystal aggregation (Ci +) is described using the Smoluchowski equation. The new element of this model is the inclusion of an additional crystal growth mechanism via the addition of primary particles (B) to crystals (Ci): (B + ). Two distinct, but constant, kernels (K not equal K') are used. It is shown that, when K' = 0, a steplike crystal size distribution (CSD) is obtained. Within a range of K'/K values (e.g., K'/K = 10(3)), CSD with multiple peaks are obtained. Comparison with predictions of models that do not include the intermediate stage of primary particles (B) indicates pronounced differences. Despite its simplicity, the model is able to capture the qualitative features of CSD evolution that have been obtained from crystal growth experiments in hematite, which is a system that is believed to undergo oriented aggregation. PMID- 16375374 TI - Near equiatomic FeCo nanocrystalline alloy embedded in an alumina aerogel matrix: microstructural features and related magnetic properties. AB - FeCo-Al2O3 nanocomposite aerogels were studied by high-resolution electron microscopy, energy filtered transmission electron microscopy, Mossbauer spectroscopy, and measurements of static magnetizations and hysteretic behavior. The combined use of such techniques provided insights on the formation of bcc FeCo nanocrystalline particles inside the alumina matrix, which is promoted by thermal treatment under hydrogen flow of the parent aerogel. Sample characteristics such as alloy composition and crystallinity, influence of the matrix on the structural evolution, and resulting magnetic properties were investigated as a function of the temperature and time of the reduction treatment. PMID- 16375375 TI - Fragmentation of the lamellae and fractionation of polymer coils upon mixing poly(dimethylacrylamide) with the lamellar phase of aerosol OT in water. AB - The lamellar mesophase formed by surfactant 1,4-bis(2-ethylhexyl) sodium sulfosuccinate (AOT) in deuterated water is mixed with poly(dimethylacrylamide) (PDMAA) polymers of low molecular weight (Mn= (2-20) x 10(3)). The mixtures separate into microphases (lamellar plus isotropic polymer solution). Their microstructures are studied by microscopy, small-angle X-ray scattering (SAXS), and deuterium NMR (2H NMR). According to SAXS, the lamellar phase fractionates the molecular weight distribution of the polymer, by dissolving only chains with coil sizes smaller than the thickness of the water layers between lamellae, and keeping larger chains segregated from the lamellar phase. The fraction of polymer that is segregated from the lamellar phase grows with Mn of the polymer. In 2H NMR, there are two signals, a quadrupolar doublet (water molecules hydrating the anisotropic lamellar phase contribute to this doublet) and a singlet (water molecules in the isotropic polymer solution contribute to this singlet). These two signals are deconvoluted to analyze the phases. Mixing with the polymer produces the partial dispersion of the lamellar phase into small fragments (microcrystallites). The structure of these microcrystallites is such that they conserve the regular long period spacing of the macrophase, and are thus identified in SAXS, but they are smaller than the minimum size required to produce quadrupolar splitting (about 4 microm), and therefore, in 2H NMR, they contribute to the singlet. 2H NMR can thus not distinguish between small microcrystallites and an isotropic polymer solution segregated from the lamellar phase; instead small microcrystallites are detected as an apparent increase of the isotropic solution. The degree of dispersion produced by the polymer in the lamellar phase is correlated with the degree of segregation that the polymer suffers. Thus, much greater dispersion into microcrystallites is produced by the higher Mn polymers than by the lower Mn polymers (in the range covered by the present samples, although with a much higher molecular weight sample (3 x 10(6)) that is totally segregated no such microcrystallites were detected). PMID- 16375376 TI - Photochemical interaction of polystyrene nanospheres with 193 nm pulsed laser light. AB - The photochemical interaction of 193 nm light with polystyrene nanospheres is used to produce particles with a controlled size and morphology. Laser fluences from 0 to 0.14 J/cm2 at 10 and 50 Hz photofragment nearly monodisperse 110 nm spherical polystyrene particles. The size distributions before and after irradiation are measured with a scanning mobility particle sizer (SMPS), and the morphology of the irradiated particles is examined with a transmission electron microscope (TEM). The results show that the irradiated particles have a smaller mean diameter ( approximately 25 nm) and a number concentration more than an order of magnitude higher than nonirradiated particles. The particles are formed by nucleation of gas-phase species produced by photolytic decomposition of nanospheres. A nondimensional parameter, the photon-to-atom ratio (PAR), is used to interpret the laser-particle interaction energetics. PMID- 16375377 TI - Molecular numbers in core and shell: structural dependence of reactivity of alkylcarboxylate-stabilized silver nanoparticles. AB - Spectroscopic, chemical, thermal, and voltammetric analyses on six kinds of alkylcarboxylate-stabilized silver nanoparticles 4.7 nm in diameter were carried out with an aim to reveal the effect of alkylcarboxylates on the optical, thermal, geometric, and electrochemical properties of the nanoparticles. These nanoparticles are composed of silver atoms and silver alkylcarboxylates having even numbers, m, of carbon atoms from 8 to 18. As a measure of the structure of the nanoparticles, the ratio of the number of silver atoms (nAg) to that of alkylcarboxylates (ns) per nanoparticle was evaluated by means of titration through chemical oxidation, voltammetric currents, and thermal gravimetric analysis. It increased with an increase in m and ranged from 1.3 to 9.8. Properties of the nanoparticle have been exhibited in absorbance of the UV-vis spectra at the point of the proportionality to n(Ag), voltammetric currents of which values were close to the theoretical values at the diffusion of particle itself, and the m-independent kinetic energy of the thermal decomposition and the overpotential of the reduction. They are not observed for the composed species, that is, silver atoms and silver alkylcarboxylate molecules. PMID- 16375378 TI - Studies of tetracene- and pentacene-based organic thin-film transistors fabricated by the neutral cluster beam deposition method. AB - The neutral cluster beam deposition (NCBD) method has been applied to produce and characterize organic thin-film transistors (OTFTs) based upon tetracene and pentacene molecules as active layers. Organic thin films were prepared by the NCBD method on hexamethyldisilazane (HMDS)-untreated and -pretreated silicon dioxide (SiO2) substrates at room temperature. The surface morphology and structures for the tetracene and pentacene thin films were examined by atomic force microscopy (AFM) and X-ray diffraction (XRD). The measurements demonstrate that the weakly bound and highly directional neutral cluster beams are efficient in producing high-quality single-crystalline thin films with uniform, smooth surfaces and that SiO2 surface treatment with HMDS enhances the crystallinity of the pentacene thin-film phase. Tetracene- and pentacene-based OTFTs with the top contact structure showed typical source-drain current modulation behavior with different gate voltages. Device parameters such as hole carrier mobility, current on/off ratio, threshold voltage, and subthreshold slope have been derived from the current-voltage characteristics together with the effects of surface treatment with HMDS. In particular, the high field-effect room-temperature mobilities for the HMDS-untreated OTFTs are found to be comparable to the most widely reported values for the respective untreated tetracene and pentacene thin film transistors. The device performance strongly correlates with the surface morphology, and the structural properties of the organic thin films are discussed. PMID- 16375379 TI - Redox properties of self-assembled gold nanoclusters. AB - The redox properties of a monolayer of alkanethiolate-protected gold nanoclusters (MPCs) constructed on a gold slide electrode was studied in 1,2-dichloroethane (DCE) electrolyte solutions. The influence of the electrostatic interaction between attached MPCs and the substrate electrode on the absolute standard redox potential of MPCs was theoretically considered and studied experimentally. PMID- 16375380 TI - Preparation and properties of ternary ZnMgO nanowires. AB - Zn0.84Mg0.16O and Zn0.12Mg0.88O nanowires with different morphology have been synthesized by a catalyst-free thermal evaporation method using Zn and Mg metals as the raw materials. X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), high-resolution transmission electron microscopy (HRTEM), and room-temperature photoluminescence (PL) measurements were used to determine the structure and optical properties of the obtained products. The obtained nanowires have diameters in a range of 30 nm-80 nm, crystallized well as hexagonal and cubic phase, with preferred orientation along the c-axis and a-axis for the two samples of Zn0.84Mg0.16O and Zn0.12Mg0.88O, respectively. Room-temperature PL at wavelengths of 384.4 and 495.8 nm has been observed for the sample of Zn0.84Mg0.16O. Upon annealing in Ar ambient, the emission peaks in PL spectra show a clearly blue shift. PMID- 16375381 TI - Gold cluster carbonyls: vibrational spectroscopy of the anions and the effects of cluster size, charge, and coverage on the CO stretching frequency. AB - We report the vibrational spectra of the carbonyl complexes of anionic gold clusters in the range of the CO stretching frequency as measured in the gas phase using IR multiple photon dissociation spectroscopy. The investigated complexes contain between 3 and 14 Au atoms and up to 7 CO ligands. Special attention is given to the complexes that exhibit saturation CO coverage as well as to the monocarbonyl species. In conjunction with data from the corresponding cationic complexes we quantify how the CO stretching frequency varies with the charge state of the gold cluster. Our results provide a size- and charge-dependent basis to interpret values of the CO stretching frequency measured for CO on deposited gold clusters in terms of the charge states of the clusters. PMID- 16375382 TI - DNA-network-templated self-assembly of silver nanoparticles and their application in surface-enhanced Raman scattering. AB - A large-scale lambda-DNA network on a mica surface was successfully fabricated with a simple method. Silver nanoparticles capped with the cationic surfactant cetyltrimethylammonium bromide (CTAB) were self-assembled onto a two-dimensional DNA network template by electrostatic interaction and formed nanoporous silver films, which can be used as active surface-enhanced raman scattering (SERS) substrates. Two probe molecules, Rhodamine 6G (R6G) and 4-aminothiophenol (4 ATP), were studied on these substrates with very low concentrations, and great enhancement factors for R6G (0.21 x 10(10)-4.09 x 10(11)) and 4-ATP (approximately 1.70 x 10(5)) were observed. It was found that the enhancement ability was affected by the DNA concentration and the electrostatic absorption time of the CTAB-stabilized silver nanoparticles on the DNA strands. These SERS substrates formed by the self-assembly of silver nanoparticles on DNA network also show good stability and reproducibility in our experiments. PMID- 16375383 TI - Chain structures of surface hydroxyl groups formed via line oxygen vacancies on TiO2(110) surfaces studied using noncontact atomic force microscopy. AB - Structures of surface hydroxyl groups arranged on a reduced TiO2(110) surface that had line oxygen vacancies were studied using noncontact atomic force microscopy (NC-AFM). NC-AFM results revealed that by increasing the density of oxygen vacancies on the TiO2(110) surface, line oxygen vacancies were formed by removal of oxygen atoms in a bridge oxygen row on the TiO2(110) surface. After the TiO2(110) surface with the line oxygen vacancies was exposed to water, the surface showed hydroxyl chain structures that were composed of hydroxyl groups linearly arranged in a form of two rows on the line oxygen vacancies and on a neighboring bridge oxygen row. In-situ NC-AFM measurements of these surfaces exposed to water at room temperature revealed that hydroxyl chain structures were formed at the line oxygen vacancy. Annealing above 500 K was sufficient to remove the hydroxyl chain structures on the TiO2(110) surface and allowed line oxygen vacancies to reappear on the surface. The line oxygen vacancies are active sites for water dissociation. In conclusion, the formation of the hydroxyl chain structure suggests that the surface hydroxyl groups on a TiO2(110) surface can be controlled by preparing oxygen vacancy structures on the surface. PMID- 16375384 TI - Orientational melting and reorientational motion in a cubane molecular crystal: a molecular simulation study. AB - Detailed molecular simulations are carried out to investigate the effect of temperature on orientational order in cubane molecular crystal. We report a transition from an orientationally ordered to an orientationally disordered plastic crystalline phase in the temperature range 425-450 K. This is similar to the experimentally reported transition at 395 K. The nature of this transition is first order and is associated with a 4.8% increase in unit cell volume that is comparable to the experimentally reported unit cell volume change of 5.4% (Phys. Rev. Lett. 1997, 78, 4938). An orientational order parameter, eta(T), has been defined in terms of average angle of libration of a molecular 3-fold axis and the orientational melting has been characterized by using eta(T). The orientational melting is associated with an anomaly in specific heat at constant pressure (C(P)) and compressibility (kappa). The enthalpy of transition and entropy of transition associated with this orientational melting are 20.8 J mol(-1) and 0.046 J mol(-1) K(-1), respectively. The structure of crystalline as well as plastic crystalline phases is characterized by using various radial distribution functions and orientational distribution functions. The coefficient of thermal expansion of the plastic crystalline phase is more than twice that of the crystalline phase. PMID- 16375385 TI - Adsorption characteristics of tripodal thiol-functionalized porphyrins on gold. AB - X-ray photoelectron and Fourier transform infrared spectroscopy studies are reported for self-assembled monolayers (SAMs) of two tripodal thiol functionalized metalloporphyrins (Zn and Cu) and three benchmark tripods on gold substrates. The tripodal unit common to all five molecules is 1-(phenyl)-1,1,1 tris(4-mercaptomethylphenyl)methane (Tpd). Both porphyrins contain S-acetyl protected thiols and are linked to the 4-position of the phenyl ring of Tpd via a phenylethyne group. The benchmark molecules include (1) two tripods containing a bromine atom at the 4-position of the apical phenyl ring, one a free thiol and the other its S-acetyl-protected analogue, and (2) a S-acetyl-protected tripod containing a phenylethyne unit at the 4-position of the apical phenyl group. Together, the spectroscopic studies reveal that none of the five tripodal molecules bond to the gold surface via all three sulfur atoms. Instead, the average number of bound thiols ranges from 1.5 to 2, with the porphyrinic molecules generally falling at the middle to upper end of the range and the smallest benchmark tripods falling at the lower end. Similar surface binding is found for the S-acetyl-protected and free benchmark tripods, indicating that the presence of the protecting group does not influence binding. Furthermore, the surface binding characteristics of the SAMs are not sensitive to deposition conditions such as solvent type, deposition time, or temperature of the solution. PMID- 16375386 TI - Thermodynamics of the N2/N3- redox couple in a LiBr-KBr-CsBr melt. AB - Nitrogen electrode reaction has been investigated in a LiBr-KBr-CsBr melt containing Li3N. The reaction N3- --> 1/2N2 + 3e- is confirmed by quantitative analysis of anodically evolved gas. The Nernst relation holds for the rest potential of Ni electrodes at a nitrogen gas pressure, pN2, of 0.05-1.0 atm and an anion fraction of the N3- ions, xN3-, of 0.003-0.010 (anion fraction). Then, the standard formal potential of the N2/N3- couple, , is evaluated to be 0.251 +/ 0.009 V versus Li+/Li (pN2 = 1 atm, xN3- = 1) at 673 K. The dependence of on the temperature (570-730 K) gives a linear relation, whose slope is (-0.930 +/- 0.117) x 10(-3) V K(-1). Thermodynamic quantities for the formation of Li3N in the melt are also estimated. PMID- 16375387 TI - Reactions of CH3SH and (CH3)2S2 on the (0001) and (000) Surfaces of ZnO. AB - Temperature-programmed desorption (TPD) was used to study the adsorption and reaction of CH3SH and (CH3)2S2 on the (0001) and (000) surfaces of ZnO. The interaction of these molecules with ZnO was found to be structure-sensitive. Both CH3SH and (CH3)2S2 adsorb dissociatively on ZnO(0001), forming adsorbed methylthiolate intermediates and only molecularly on ZnO(000). In the case of CH3SH, this result is consistent with that reported previously for the interaction of Bronsted acids with ZnO and indicates that exposed cation-anion pairs are the active sites for dissociative adsorption. Only exposed Zn cations are required for the dissociative adsorption of (CH3)2S2. Methylthiolate species produced by dissociative adsorption of CH3SH and (CH3)2S2 on ZnO(0001) were found to undergo a variety of reaction pathways, including coupling to produce dimethyl sulfide and oxidation to formaldehyde, CO, and CO2. Pathways for the production of these various products are proposed. PMID- 16375388 TI - Ab initio study on adsorption of hydrated Na+ and Cu+ cations on the Cu(111) surface. AB - The interactions of Na+ and Cu+ cations with a Cu(111) surface in the presence and absence of water molecules were investigated using cluster models and ab initio methods. Adsorption in aqueous solution was modeled with one to five water molecules around the adsorbing cation. The Cu surface was described with Cu10 and Cu18 cluster models and the computational method was MP2/RECP/6-31+G. The effect of the basis set superposition error (BSSE) was taken into account with counterpoise (CP) correction, and the accuracy of HF-level results was examined. The interactions between Na+ and the Cu surface were found to be primarily electrostatic, and the energy differences among the different adsorption sites were small. The largest CP-corrected MP2 adsorption energy for the Cu18 cluster was -188 kJ/mol. When water molecules were added around it, Na+ receded from the Cu surface and finally was surrounded totally by the water molecules. The interactions between Cu+ and the Cu surface were dominated by orbital interactions, and Cu+ preferred to adsorb on sites where it could bind to more than one surface atom. The largest CP-corrected MP2 adsorption energy for the Cu18 cluster was -447 kJ/mol. Adding water molecules around it did not cause Cu+ to draw away from the surface, but instead the water molecules began to form hydrogen bonds with one another. The magnitude of BSSE was substantial in most cases. CP corrections did not, however, have a significant impact on the relative trends among the interaction energies. PMID- 16375389 TI - Destructive adsorption of CCl4 over lanthanum-based solids: linking activity to acid-base properties. AB - The relative activities of a low-surface crystalline and high-surface amorphous LaOCl, further denoted as S1 and S2, have been compared for the destructive adsorption of CCl4. It was found that the intrinsic activity of S2 is higher than that of S1. Both samples were characterized with X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), N2-physisorption, and Raman and infrared (IR) spectroscopy. IR was used in combination with CO2, CO, and methanol as probe molecules. The CO2 experiments showed that different carbonate species are formed on both materials. For S1, a high surface concentration of bidentate carbonate species and a lower concentration of monodentate carbonate were observed. In the case of S2, bulk carbonates were present together with bridged carbonates. CO adsorption shows that S2 and S1 have very similar Lewis acid sites. However, methanol adsorption experiments showed that S2 had a higher number of stronger Lewis acid sites than S1 and that twofold coordinated methoxy species were more strongly bound than threefold coordinated methoxy species. Because of the analogy between methanol dissociation and the removal of the first chlorine atom in the destructive adsorption of CCl4, the sites enabling twofold coordination were likely to be the same Lewis acid sites actively involved in the destructive adsorption of CCl4. La2O3 was less active than the two LaOCl materials, and therefore, the intrinsic activity of the catalyst increases as the strength of the Lewis acid sites increases. S2 contains more chlorine at the surface than S1, which is expressed by the higher number of sites enabling twofold coordination. Moreover, this explains the difference in destructive adsorption capacity for CCl4 that was observed for the samples S1 and S2. Since LaCl3, being the most acidic phase, is not active for the destructive adsorption of CCl4, basic oxygen atoms, however, remain needed to stabilize the reaction intermediate CCl3 as La-O CCl3. PMID- 16375390 TI - IR chemiluminescence probe of the vibrational energy distribution of CO2 formed during steady-state CO oxidation on Pt(111) and Pt(110) surfaces. AB - The infrared (IR) chemiluminescence spectra of CO2 were measured during the steady-state CO + O2 reaction over Pt(110) and Pt(111) surfaces. Analysis of the IR emission spectra indicates that the bending vibrational temperature (TVB), as well as the antisymmetric vibrational temperature (TVAS), was higher on Pt(110) than on Pt(111). On the Pt(110) surface, the highly excited bending vibrational mode compared to the antisymmetric vibrational mode was observed under reaction conditions at low CO coverage (theta(CO) < 0.2) or at high surface temperatures (TS > or = 700 K). This can be related to the activated complex of CO2 formation in a more bent form on the inclined (111) terraces of the Pt(110)(1 x 2) structure. On the other hand, at high CO coverage (theta(CO) > 0.2) or at low surface temperatures (TS < 650 K), TVAS was higher than TVB, which can be caused by the reconstruction of the Pt(110)(1 x 2) surface to the (1 x 1) form with high CO coverage. PMID- 16375391 TI - Kinetic Monte Carlo simulations of nucleation and growth in electrodeposition. AB - Nucleation and growth during bulk electrodeposition is studied using kinetic Monte Carlo (KMC) simulations. Ion transport in solution is modeled using Brownian dynamics, and the kinetics of nucleation and growth are dependent on the probabilities of metal-on-substrate and metal-on-metal deposition. Using this approach, we make no assumptions about the nucleation rate, island density, or island distribution. The influence of the attachment probabilities and concentration on the time-dependent island density and current transients is reported. Various models have been assessed by recovering the nucleation rate and island density from the current-time transients. PMID- 16375392 TI - Pneumatochemical impedance spectroscopy. 1. Principles. AB - Pneumatochemical impedance spectroscopy (PIS) is the indirect transposition of electrochemical impedance spectroscopy (EIS) to solid-gas reactions. In PIS analysis, an analogy is made between pressure and electrical potential on one hand and gas flow and electric current on the other hand, and pneumatochemical transfer functions are derived from gas-phase measurements. Potentially, this spectroscopy can be used to analyze the dynamics of any solid-gas system including adsorption (surface) and absorption (bulk) phenomena, gas (H2) permeation across metallic membranes, and electrocatalysis in gaseous fuel cells. Hydrogen absorption by intermetallic compounds (IMCs), a process of great practical interest for hydrogen storage applications, is more specifically considered in this paper, and the kinetic equations derived in this work pertain only to this case. Whereas classical electrochemical impedance measurements are performed using an harmonic analyzer and monochromatic potential (potentiostatic mode) or current (intentiostatic mode) perturbations, PIS investigation of the dynamics of IMC-H2(g) systems is more conveniently performed using Sievert's-type gas distribution apparatus (SGDA) and polychromatic pressure perturbation signals. This is first because monochromatic isothermal pressure modulations cannot be easily obtained experimentally over the frequency domain of interest and, second, because most IMC-H2(g) systems exhibit strongly nonlinear behaviors in two-phase domains (hysteresis), and this proscribes harmonic analysis. A further benefit of using SGDA and nonharmonic perturbations is that kinetic and thermodynamic information are collected simultaneously during the same experiment. The measurement and modeling of the pneumatochemical transfer functions associated with IMC-H2(g) systems, both in solid solution and two-phase domains, are discussed in this paper which is organized in two parts. The principles of PIS analysis, based on the theory of linear and time-invariant systems, are presented in the first part. The dynamics of hydrogen sorption by metals and IMCs is analyzed in the second part, where a detailed analysis of the multistep reaction paths involved in sorption mechanisms is proposed. PMID- 16375393 TI - Pneumatochemical impedance spectroscopy. 2. Dynamics of hydrogen sorption by metals. AB - In this paper, pneumatochemical impedance spectroscopy is used to analyze multistep reaction mechanisms such as those observed in solid solution domains of LaNi5-H2(g) systems. It is shown that hydrogen sorption is a two-step mechanism including (i) dissociative surface chemisorption of molecular hydrogen and (ii) atomic hydrogen bulk transport by diffusion. Data fitting of experimental transfer functions with model equations yields the value of the kinetic parameter associated with each individual reaction step, i.e., surface sorption resistances and hydrogen bulk diffusion coefficients. The technique is used to follow the activation procedure of the sample as well as the degradation of sorption properties in oxygen-containing hydrogen atmospheres. A decrease in sorption kinetics is attributed to surface oxidation, whereas bulk properties remain unchanged. The perspectives offered by the technique which potentially can be used to optimize surface and bulk composition of IMC for increased sorption rates are discussed. PMID- 16375394 TI - Deformation of a "rigid" molecule in self-assembled nanostructures. AB - A simple spirobifluorene molecule with pseudotetrahedral structure was investigated for its supposed conformational resilience upon adsorption. Through deposition at room temperature of this molecule on a Cu(111) surface and subsequent observation at 5 K with an ultrahigh vacuum scanning tunneling microscope, this "rigidity" upon physisorption is confirmed. However, an unexpected chemisorbed state was also found with the molecules arranged in trimers. The unique coexistence of physisorbed and chemisorbed states on the same substrate is thus demonstrated at the early stage of self-assembly. PMID- 16375395 TI - A kinetic model of the formation of organic monolayers on hydrogen-terminated silicon by hydrosilation of alkenes. AB - We have analyzed a kinetic model for the formation of organic monolayers based on a previously suggested free radical chain mechanism for the reaction of unsaturated molecules with hydrogen-terminated silicon surfaces (Linford, M. R.; Fenter, P. M.; Chidsey, C. E. D. J. Am. Chem. Soc 1995, 117, 3145). A direct consequence of this mechanism is the nonexponential growth of the monolayer, and this has been observed spectroscopically. In the model, the initiation of silyl radicals on the surface is pseudo first order with rate constant, ki, and the rate of propagation is determined by the concentration of radicals and unreacted Si-H nearest neighbor sites with a rate constant, kp. This propagation step determines the rate at which the monolayer forms by addition of alkene molecules to form a track of molecules that constitute a self-avoiding random walk on the surface. The initiation step describes how frequently new random walks commence. A termination step by which the radicals are destroyed is also included. The solution of the kinetic equations yields the fraction of alkylated surface sites and the mean length of the random walks as a function of time. In mean-field approximation we show that (1) the average length of the random walk is proportional to (kp/ki)1/2, (2) the monolayer surface coverage grows exponentially only after an induction period, (3) the effective first-order rate constant describing the growth of the monolayer and the induction period (kt) is k = (2ki kp)1/2, (4) at long times the effective first-order rate constant drops to ki, and (5) the overall activation energy for the growth kinetics is the mean of the activation energies for the initiation and propagation steps. Monte Carlo simulations of the mechanism produce qualitatively similar kinetic plots, but the mean random walk length (and effective rate constant) is overestimated by the mean field approximation and when kp >> ki, we find k approximately ki0.7kp0.3 and Ea = (0.7Ei+ 0.3Ep). However the most striking prediction of the Monte Carlo simulations is that at long times, t >> 1/k, the effective first-order rate constant decreases to ki even in the absence of a chemical termination step. Experimental kinetic data for the reaction of undec-1-ene with hydrogen terminated porous silicon under thermal reflux in toluene and ethylbenzene gave a value of k = 0.06 min(-1) and an activation energy of 107 kJ mol(-1). The activation energy is in reasonable agreement with density functional calculations of the transition state energies for the initiation and propagation steps. PMID- 16375396 TI - Roles of surface Te, Nb, and Sb oxides in propane oxidation to acrylic acid over bulk orthorhombic Mo-V-O phase. AB - The outermost surfaces and subsurface layers of the orthorhombic (M1) Mo-V-O catalysts promoted with Te, Nb, and Sb oxide species at submonolayer surface coverage were examined by low-energy ion scattering (LEIS). This study indicated that the Nb oxide species was preferentially located at the topmost surface, while the subsurface Te and Sb concentrations declined gradually into the bulk. Although the original Mo-V-O catalyst was essentially unselective in propane oxidation to acrylic acid, significant improvement in the selectivity to acrylic acid was observed when Te, Nb, and Sb oxides were present as the surface species at submonolayer coverage. These findings further suggested that the formation of the surface V-O-M bonds (M = Nb, Te, or Sb) was highly beneficial for both the activity and selectivity of the orthorhombic Mo-V-O catalysts in propane oxidation to acrylic acid. The highest selectivity was observed when both Nb and Te (or Sb) oxide species were present at the surface. The selectivity trends established for the surface-promoted Mo-V-O catalyst parallel those found previously for the corresponding bulk Mo-V-M-O catalysts. These results further indicated that the introduction of surface metal oxide species is a highly promising method to prepare well-defined model catalysts for studies of the structure-activity/selectivity relationships as well as optimize the catalytic performance of the bulk mixed Mo-V-M-O catalysts for selective (amm)oxidation of propane. PMID- 16375397 TI - Surfaces affect ion pairing. AB - In water, positive ions attract negative ions. That attraction can be modulated if a hydrophobic surface is present near the two ions in water. Using computer simulations with explicit and implicit water, we study how an ion embedded on a hydrophobic surface interacts with another nearby ion in water. Using hydrophobic surfaces with different curvatures, we find that the contact interaction between a positive and negative ion is strongly affected by the curvature of an adjacent surface, either stabilizing or destabilizing the ion pair. We also find that the solvent-separated ion pair (SSIP) can be made more stable than the contacting ion pair by the presence of a surface. This may account for why bridging waters are often found in protein crystal structures. We also note that implicit solvent models do not account for SSIPs. Finally, we find that there are charge asymmetries: an embedded positive charge attracting a negative ion is different than an embedded negative charge attracting a positive ion. Such asymmetries are also not predicted by implicit solvent models. These results may be useful for improving computational models of solvation in biology and chemistry. PMID- 16375398 TI - Modeling the cavitation free energy. AB - A new expression to compute the cavitation free energy has been derived by integrating a new model to fit its derivative with respect to the cavity radius. The derivatives were obtained from Monte Carlo simulations data of the contact values of distribution functions for hard-sphere solutes in TIP4P water at 298 K and 1 atm. The new expression, formulated in the framework of the thermodynamics of surfaces and unlike the classical simple models, gives good results also for very small cavities with a radius of approximately 1 A. The contribution to the free energy of a term, which depends on the excess number of molecules at the dividing surface, has been taken into account and discussed for the assumed dependence on r of the surface tension. The asymptotic behavior of the derivative has thus been considered, and a function t(r), which is 0 at r = 0 and 1 at infinity, has been introduced to describe the transition from small to large length regimes. The value of the surface tension obtained by fitting is in very good agreement with that obtained from a simulation of the liquid/vapor interface by using the TIP4P model. PMID- 16375399 TI - Water condensation in hydrophobic silicalite-1 zeolite: a molecular simulation study. AB - We report grand canonical Monte Carlo simulations of the gas and liquid phase adsorption of water in silicalite-1 zeolite. Simple but effective models and simulation methods, found useful for studying gas adsorption in nanoporous materials, have been extended to describe the intrusion/extrusion cycle of water in this hydrophobic solid. The picture of water confined to hydrophobic spaces of nanoscopic dimensions that emerges from this study is one of a strongly depleted and highly inhomogeneous fluid. PMID- 16375400 TI - Phase and glass transitions in short-range central potential model systems: the case of C60. AB - Extensive molecular dynamics simulations show that a short-range central potential, suited to model C60, undergoes a high temperature transition to a glassy phase characterized by the positional disorder of the constituent particles. Crystallization, melting, and sublimation, which also take place during the simulation runs, are illustrated in detail. It turns out that vitrification and the mentioned phase transitions occur when the packing fraction of the system-defined in terms of an effective hard-core diameter-equals that of hard spheres at their own glass and melting transition, respectively. A close analogy also emerges between our findings and recent mode coupling theory calculations of structural arrest lines in a similar model of protein solutions. We argue that the conclusions of the present study might hold for a wide class of potentials currently employed to mimic interactions in complex fluids (some of which are of biological interest), suggesting how to achieve at least qualitative predictions of vitrification and crystallization in those systems. PMID- 16375401 TI - Effects of molecular association on polarizability relaxation in liquid mixtures of benzene and hexafluorobenzene. AB - In this work we have studied the relaxation dynamics of the many-body polarizability anisotropy in liquid mixtures of benzene (Bz) and hexafluorobenzene (Hf) at room temperature by femtosecond optical heterodyne detected Raman-induced Kerr effect spectroscopy (OHD-RIKES) experiments and molecular dynamics (MD) simulations. The computed polarizability response arising from intermolecular interactions was included using the first-order dipole induced-dipole model with the molecular polarizability distributed over the carbon sites of each molecule. We found good qualitative agreement between experiments and simulations in the features exhibited by the nuclear response function R(t) for pure liquids and mixtures. The long-time diffusive decay of R(t) was observed to vary substantially with composition, slowing down noticeably with dilution of each of the species as compared with that in the corresponding pure liquids. MD simulation shows that the effect on R(t) is due to the formation of strong and localized intermolecular association between Bz and Hf species that hinder the rotational diffusive dynamics. The formation of these Bz-Hf complexes in the liquid mixtures also modifies the rotational diffusive dynamics of the component species in such a way that cannot be explained solely in terms of a viscosity effect. Even though the computed orientational diffusive relaxation times associated with Bz and Hf are larger by a factor of approximately 2 than those from experiments, we found similar trends in experiments and simulations for these characteristic times as a function of composition. Namely, the collective and single-molecule orientational correlation times associated with Bz are observed to grow monotonically with the dilution of Bz, while those corresponding to Hf species exhibit a maximum at the equimolar composition. We attribute the quantitative discrepancy between experiments and simulations to the use of the Williams potential, which seems to overestimate the intermolecular interactions and thus predicts not only a slower translational dynamics but also a slower rotational diffusion dynamics than in real fluids. PMID- 16375402 TI - Transferable potentials for phase equilibria. 8. United-atom description for thiols, sulfides, disulfides, and thiophene. AB - An extension of the transferable potentials for phase equilibria united-atom (TraPPE-UA) force field to thiol, sulfide, and disulfide functionalities and thiophene is presented. In the TraPPE-UA force field, nonbonded interactions are governed by a Lennard-Jones plus fixed point charge functional form. Partial charges are determined through a CHELPG analysis of electrostatic potential energy surfaces derived from ab initio calculations at the HF/6-31g+(d,p) level. The Lennard-Jones well depth and size parameters for four new interaction sites, S (thiols), S(sulfides), S(disulfides), and S(thiophene), were determined by fitting simulation data to pure-component vapor-equilibrium data for methanethiol, dimethyl sulfide, dimethyl disulfide, and thiophene, respectively. Configurational-bias Monte Carlo simulations in the grand canonical ensemble combined with histogram-reweighting methods were used to calculate the vapor liquid coexistence curves for methanethiol, ethanethiol, 2-methyl-1-propanethiol, 2-methyl-2-propanethiol, 2-butanethiol, pentanethiol, octanethiol, dimethyl sulfide, diethyl sulfide, ethylmethyl sulfide, dimethyl disulfide, diethyl disulfide, and thiophene. Excellent agreement with experiment is achieved, with unsigned errors of less than 1% for saturated liquid densities and less than 3% for critical temperatures. The normal boiling points were predicted to within 1% of experiment in most cases, although for certain molecules (pentanethiol) deviations as large as 5% were found. Additional calculations were performed to determine the pressure-composition behavior of ethanethiol+n-butane at 373.15 K and the temperature-composition behavior of 1-propanethiol+n-hexane at 1.01 bar. In each case, a good reproduction of experimental vapor-liquid equilibrium separation factors is achieved; both of the coexistence curves are somewhat shifted because of overprediction of the pure-component vapor pressures. PMID- 16375403 TI - Ultrasonic velocities, densities, viscosities, electrical conductivities, Raman spectra, and molecular dynamics simulations of aqueous solutions of Mg(OAc)2 and Mg(NO3)2: Hofmeister effects and ion pair formation. AB - The ultrasonic velocities, densities, viscosities, and electrical conductivities of aqueous solutions of magnesium nitrate and magnesium acetate have been measured from dilute to saturation concentrations at 0 < or = t/degrees C < or = 50. The temperature derivative of the isentropic compressibility, kappa(s), became zero at 2.28 and 2.90 mol kg(-1) for Mg(OAc)2 and Mg(NO3)2 solutions, respectively, at 25 degrees C. The total hydration numbers of the dissolved ions were estimated to be, respectively, 24.3 and 19.2 at these concentrations. Differences in kappa(s) for various M2+ salts, using the present and literature data, correlated with reported M2+-OH2 bond lengths and to a lesser extent with cationic charge densities (ionic radii). The influence of anions on kappa(s) appears to follow the Hofmeister series and also correlates approximately with the anionic charge density. Substantial differences between Mg(OAc)2(aq) and Mg(NO3)2(aq) occur with respect to their structural relaxation times (derived from compressibility and viscosity data) and their electrical conductivities. These differences were attributed to a much greater ion association in Mg(OAc)2 solutions. Raman spectra recorded at 28 degrees C confirmed the presence of various types of contact ion pairs including mono- and bidentate complexes in Mg(OAc)2(aq). In Mg(NO3)2(aq), only noncontact ion pairs appear to be formed even at high concentrations. The experimental results are supported by molecular dynamics simulations, which also reveal the much stronger tendency of OAc- compared to NO3- to associate with Mg2+ in aqueous solutions. The simulations also allow an evaluation of the ion-ion and ion-water radial distribution functions and cumulative sums and provide a molecular picture of ion hydration in Mg(OAc)2(aq) and Mg(NO3)2(aq) at varying concentrations. PMID- 16375404 TI - Cyan fluorescent protein: molecular dynamics, simulations, and electronic absorption spectrum. AB - The dynamics and electronic absorption spectrum of enhanced cyan fluorescent protein (ECFP), a mutant of green fluorescent protein (GFP), have been studied by means of a 1 ns molecular dynamics (MD) simulation. The two X-ray conformations A' and B' of ECFP were considered. The chromophore was assumed to be neutral, and all titratable residues were taken in their standard protonation state at neutral pH. The protein was embedded in a box of water molecules (and counterions). The first result is that the two conformations A' and B' are found to be stable all along the simulation. Then, an analysis of the hydrogen-bond networks shows strong differences between the two conformations in the surroundings of the nitrogen atom of the indolic part of the chromophore. This is partly due to the imperfection in the beta barrel near the His148 residue, which allows the access of one solvent molecule inside the protein in conformation A'. Finally, quantum mechanical calculations of the electronic transition energies of the chromophore in the charge cloud of the protein and solvent water molecules were performed using the TDDFT method on 160 snapshots extracted every 5 ps of the MD trajectories. It is found that conformations A' and B' exhibit very similar spectra despite different H-bond networks involving the chromophore. This similarity is related to the weak charge transfer involved in the electronic transition and the weak electrostatic field created by ECFP near the chromophore, within the hypotheses made in the present simulation. PMID- 16375405 TI - Relation between solvent and protein dynamics as studied by dielectric spectroscopy. AB - We present results obtained by dielectric spectroscopy in wide frequency (10(-2) 10(9) Hz) and temperature ranges on human hemoglobin in the three different solvents water, glycerol, and methanol, at a solvent level of 0.8 g of solvent/g of protein. In this broad frequency region, there are motions on several time scales in the measured temperature range (110-370 K for water, 170-410 K for glycerol, and 110-310 K for methanol). For all samples, the dielectric data shows at least four relaxation processes, with frequency dependences that are well described by the Havriliak-Negami or Cole-Cole functions. The fastest and most pronounced process in the dielectric spectra of hemoglobin in glycerol and methanol solutions is similar to the alpha-relaxation of the corresponding bulk solvent (but shifted to slower dynamics due to surface interactions). For water solutions, however, this process corresponds to earlier results obtained for water confined in various systems and it is most likely due to a local beta relaxation. The slowing down of the glycerol and methanol relaxations and the good agreement with earlier results on confined water show that this process is affected by the interaction with the protein surface. The second fastest process is attributed to motions of polar side groups on the protein, with a possible contribution from tightly bound solvent molecules. This process is shifted to slower dynamics with increasing solvent viscosity, and it shows a crossover in its temperature dependence from Arrhenius behavior at low temperatures to non Arrhenius behavior at higher temperatures where there seems to be an onset of cooperativity effects. The origins of the two slowest relaxation processes (visible at high temperatures and low frequencies), which show saddlelike temperature dependences for the solvents water and methanol, are most likely due to motions of the polypeptide backbone and an even more global motion in the protein molecule. PMID- 16375406 TI - Interactions of trimethylamine N-oxide and water with cyclo-alanylglycine. AB - The osmolyte trimethylamine N-oxide (TMAO) is one of a family of compounds found in living systems that can stabilize biomolecular tertiary structures. As a step in exploring the interactions between this material and polyamino acids, we have determined intermolecular 1H{1H} nuclear Overhauser effects (NOEs) between the protons of cyclo-alanylglycine and protons of solvent components in TMAO-water solutions. Comparison of the results to effects predicted on the basis of the molecular shape of the dipeptide and experimental translational diffusion coefficients suggests that both water and TMAO molecules have properties in the vicinity of the dipeptide that are different from those in the bulk solution. Changes of local concentrations of water and TMAO and changes in the diffusive behavior of these components near the dipeptide are rejected as possible explanations of the discrepancies between observed and calculated Overhauser effects. Rather, it is concluded that TMAO molecules, and the water molecules associated with them, participate to some extent in the formation of long-lived solute-solvent complexes. The aliphatic alcohol tert-butyl alcohol is structurally similar to TMAO. Overhauser effect studies of its interaction with cyclo-alanylglycine in tert-butyl alcohol-water suggest similar kinds of interactions are present in this system but that they are significantly weaker, presumably because of the lower polarity of this alcohol compared to TMAO. PMID- 16375407 TI - Carbohydrate intramolecular hydrogen bonding cooperativity and its effect on water structure. AB - Molecular dynamics (MD) simulations combined with water-water H-bond angle analysis and calculation of solvent accessible surface area and approximate free energy of solvation were used to determine the influence of hydroxyl orientation on solute hydration and surrounding water structure for a group of chemically identical solutes-the aldohexopyranose sugars. Intramolecular hydrogen bond cooperativity was closely associated with changes in water structure surrounding the aldohexopyranose stereoisomers. The OH-4 group played a pivotal role in hydration as it was able to participate in a number of hydrogen bond networks utilizing the OH-6 group. Networks that terminated within the molecule (OH-4 --> OH-6 --> O-5) had relatively more nonpolar-like hydration than those that ended in a free hydroxyl group (OH-6 --> OH-4 --> OH-3). The OH-2 group modulated the strength of OH-4 networks through syndiaxial OH-2/4 intramolecular hydrogen bonding, which stabilized and induced directionality in the network. Other syndiaxial interactions, such as the one between OH-1 and OH-3, only indirectly affected water structure. Water structure surrounding hydrogen bond networks is discussed in terms of water-water hydrogen bond populations. The impact of syndiaxial versus vicinal hydrogen bonds is also reviewed. The results suggest that biological events such as protein-carbohydrate recognition and cryoprotection by carbohydrates may be driven by intramolecular hydrogen bond cooperativity. PMID- 16375408 TI - Probing the contribution of electronic coupling to the directionality of electron transfer in photosynthetic reaction centers. AB - Subpicosecond transient absorption studies are reported for a set of Rhodobacter (R.) capsulatus bacterial photosynthetic reaction centers (RCs) designed to probe the origins of the unidirectionality of charge separation via one of two electron transport chains in the native pigment-protein complex. All of the RCs have been engineered to contain a heterodimeric primary electron donor (D) consisting of a bacteriochlorophyll (BChl) and a bacteriopheophytin (BPh). The BPh component of the M heterodimer (Mhd) or L heterodimer (Lhd) is introduced by substituting a Leu for His M200 or His L173, respectively. Previous work on primary charge separation in heterodimer mutants has not included the Lhd RC from R. capsulatus, which we report for the first time. The Lhd and Mhd RCs are used as controls against which we assess RCs that combine the heterodimer mutations with a second mutation (His substituted for Leu at M212) that results in replacement of the native L-side BPh acceptor with a BChl (beta). The transient absorption spectra reveal clear evidence for charge separation to the normally inactive M-side BPh acceptor (H(M)) in Lhd-beta RCs to form D+H(M)- with a yield of approximately 6%. This state also forms in Mhd-beta RCs but with about one-quarter the yield. In both RCs, deactivation to the ground state is the predominant pathway of D decay, as it is in the Mhd and Lhd single mutants. Analysis of the results indicates an upper limit ofV2L/V2m < or = 4 for the contribution of the electronic coupling elements to the relative rates of electron transfer to the L versus M sides of the wild-type RC. In comparison to the L/M rate ratio (kL/kM) approximately 30 for wild-type RCs, our findings indicate that electronic factors contribute approximately 35% at most to directionality with the other 65% deriving from energetic considerations, which includes differences in free energies, reorganization energies, and contributions of one- and two-step mechanisms on the two sides of the RC. PMID- 16375409 TI - Modulating membrane properties: the effect of trehalose and cholesterol on a phospholipid bilayer. AB - The protective properties of trehalose on cholesterol-containing lipid dipalmitoylphosphatidylcholine (DPPC) bilayers are studied through molecular simulations. The ability of the disaccharide to interact with the phospholipid headgroups and stabilize the membrane persists even at high cholesterol concentrations and restricts some of the changes to the structure that would otherwise be imposed by cholesterol molecules. Predictions of bilayer properties such as area per lipid, tail ordering, and chain conformation support the notion that the disaccharide decreases the main melting transition in these multicomponent model membranes, which correspond more closely to common biological systems than pure bilayers. Molecular simulations indicate that the membrane dynamics are slowed considerably by the presence of trehalose, indicating that high sugar concentrations would serve to avert possible phase separations that could arise in mixed phospholipid systems. Various time correlation functions suggest that the character of the modifications in lipid dynamics induced by trehalose and cholesterol is different in the hydrophilic and hydrophobic regions of the membrane. PMID- 16375410 TI - Charging/discharging of Au (core)/silica (shell) nanoparticles as revealed by XPS. AB - By recording XPS spectra while applying external voltage stress to the sample rod, we can control the extent of charging developed on core-shell-type gold nanoparticles deposited on a copper substrate, in both steady-state and time resolved fashions. The charging manifests itself as a shift in the measured binding energy of the corresponding XPS peak. Whereas the bare gold nanoparticles exhibit no measurable binding energy shift in the Au 4f peaks, both the Au 4f and the Si 2p peaks exhibit significant and highly correlated (in time and magnitude) shifts in the case of gold (core)/silica (shell) nanoparticles. Using the shift in the Au 4f peaks, the capacitance of the 15-nm gold (core)/6-nm silica (shell) nanoparticle/nanocapacitor is estimated as 60 aF. It is further estimated that, in the fully charged situation, only 1 in 1000 silicon dioxide units in the shell carries a positive charge during our XPS analysis. Our simple method of controlling the charging, by application of an external voltage stress during XPS analysis, enables us to detect, locate, and quantify the charges developed on surface structures in a completely noncontact fashion. PMID- 16375411 TI - Nanometer-scale ion aggregates in aqueous electrolyte solutions: guanidinium sulfate and guanidinium thiocyanate. AB - Neutron diffraction experiments and molecular dynamics simulations are used to study the structure of aqueous solutions of two electrolytes: guanidinium sulfate (a mild protein conformation stabilizer) and guanidinium thiocyanate (a powerful denaturant). The MD simulations find the unexpected result that in the Gdm2SO4 solution the ions aggregated into mesoscopic (nanometer-scale) clusters, while no such aggregation is found in the GdmSCN solution. The neutron diffraction studies, the most direct experimental probe of solution structure, provide corroborating evidence that the predicted very strong ion pairing does occur in solutions of 1.5 m Gdm2SO4 but not in 3 m solutions of GdmSCN. A mechanism is proposed as to how this mesoscopic solution structure affects solution denaturant properties and suggests an explanation for the Hofmeister ordering of these solutions in terms of this ion pairing and the ability of sulfate to reverse the denaturant power of guanidinium. PMID- 16375412 TI - Radical scavenging and catalytic activity of metal-phenolic complexes. AB - A series of metal-ligand complexes were prepared by the reaction of various metal ions, namely, Cu(II), Mn(II), or Fe(II) with phenolic derivatives of [catechol, chlorogenic acid (CGA), n-propyl gallate (nPG), 3-hydroxy anthranilic acid, resveratrol, and rutin] and characterized by UV-vis spectroscopy. The metal/ligand complexing ratio and complexation constants have been determined. The complexes were probed for their reactivity toward various free radicals (e aq , CO2*-, and O2*-). Pulse radiolysis studies showed that the one-electron reduction of metal/phenol complexes by CO2*- radicals was metal-centered, and this was confirmed by the formation of an initial adduct with CO2*- radicals. Rate constants for the scavenging of superoxide anions with metal complexes ranged between 10(7)-10(9) dm3 mol(-1) s(-1) and those for the reaction of e aq- with the metal complexes were in the range of (1-5) x 10(9) dm3 mol(-1) s(-1), depending on the pH of the solution. Cyclic and differential pulse voltammetric studies showed that the reduction potential of the complexes are found to range between -0.022 to 0.45 V vs normal hydrogen electrode. PMID- 16375413 TI - Enhancing the rate of the Diels-Alder reaction using CO2 + ethanol and CO2 + n hexane mixed solvents of different phase regions. AB - The reaction rate of the Diels-Alder reaction between N-ethylmaleimide and 9 hydroxymethylanthrance in CO2 + ethanol and CO2 + hexane mixed solvents of different compositions were determined by in situ UV/vis spectroscopy at 318.15 K and different pressures. The density of the mixed solvents at different pressures was also determined and the isothermal compressibility was calculated using the density data. The activation volume of the reaction was calculated based on the dependence of rate constant (kc) on pressure. It was demonstrated that the kc was very sensitive to the pressure in the mixed solvents near the critical region and the kc increased dramatically as pressure approached dew points, critical point, and bubble points of the mixed solvents. However, the kc in the mixed solvents outside the critical region or in pure CO2 was not sensitive to pressure. At suitable conditions, kc could be 40 times larger than that in acetonitrile. The activation volume of the reaction was nearly independent of pressure as the pressure was much higher than the phase separation pressure of the mixed solvents, while it increased considerably as pressure approached the bubble points, critical point, and dew points from high pressure. The clustering of the solvent molecules with the reactants and the activated complex in the reaction systems near the phase boundary in the critical region may be the main reason for the interesting phenomena observed. This work also shows that, using pure CO2 as the solvent, the reaction cannot be carried out in the critical region of the solvent due to the limitations of the reactants, while it can be conducted in the critical region of mixed solvents of suitable compositions, where the solvents are highly compressible and the reaction rate can be tuned effectively by pressure. PMID- 16375414 TI - Interface-specific chi(4) coherent Raman spectroscopy in the frequency domain. AB - We demonstrate interface-specific fourth-order (chi(4)) coherent Raman spectroscopy in the frequency domain for the first time. Because the chi(4) Raman spectroscopy uses only visible (vis) or near-IR light, it is expected to be a potential alternative to the widely utilized IR-vis sum frequency generation spectroscopy that cannot be applied to interfaces buried in thick IR absorbers such as water. We present the vibrational absolute value(chi(4))2 spectrum of rhodamine 800 at the air/water interface in a wide spectral range 100-3600 cm( 1). Comparison of the absolute value(chi(4))2 spectrum with the absolute value(chi(3))2 spectrum leads us to conclude that the present chi(4) spectroscopy successfully probes the interface distinguished clearly from the bulk. PMID- 16375415 TI - Iron silicide root formation in carbon nanotubes grown by microwave PECVD. AB - Aligned carbon nanotubes have been grown using microwave plasma enhanced chemical vapor deposition (PECVD). The carbon nanotubes are nucleated from iron catalyst particles which, during growth, remain adherent to the silicon substrates. By analysis with high-resolution electron microscopy, we observe iron silicide roots penetrating into the silicon substrate at the interface of the catalyst particles and the substrate, thus providing strong adhesion of the carbon nanotubes onto the substrate. The iron silicide roots assist in the attachment of the catalyst particles to the substrate and play a role in the evolution of the catalyst particle morphology and resulting base growth mode. Carbon nanotubes grown by microwave PECVD could exhibit superior electrical and thermal transport properties over other PECVD processes, so an understanding of the growth mechanism is important for utilization in device applications. PMID- 16375416 TI - Observation of individual microtubule motor steps in living cells with endocytosed quantum dots. AB - We report the observation of individual steps taken by motor proteins in living cells by following movements of endocytic vesicles that contain quantum dots (QDs) with a fast camera. The brightness and photostability of quantum dots allow us to record motor displacement traces with 300 micros time resolution and 1.5 nm spatial precision. We observed individual 8 nm steps in active transport toward both the microtubule plus- and minus-ends, the directions of kinesin and dynein movements, respectively. In addition, we clearly resolved abrupt 16 nm steps in the plus-end direction and often consecutive 16 nm and occasional 24 nm steps in minus-end directed movements. This work demonstrates the ability of the QD assay to probe the operation of motor proteins at the molecular level in living cells under physiological conditions. PMID- 16375417 TI - The interplay of hydrophobic and electrostatic effects in the surfactant-induced aggregation/deaggregation of chlorin p6. AB - The aggregation/deaggregation of chlorin p6 with the surfactants CTAB, SDS, and TX 100 have been studied by using absorption, fluorescence, and light scattering techniques. The ionic surfactants are found to cause aggregation of fluorophore at submicellar concentrations. The aggregates dissolve at higher surfactant concentrations to yield micellized monomers. This is rationalized by the interplay of electrostatic and hydrophobic effects. A prominent pH effect is observed in the ionic surfactant induced aggregation process as the charge on the fluorophore is controlled by the pH of the medium. Interestingly, the neutral TX 100 also induces aggregation of chlorin p6 at low concentrations, indicating that hydrophobic effects by themselves can cause aggregation unless there is a hindrance by repulsive electrostatic effects. PMID- 16375418 TI - Architecture of a biocompatible supramolecular material by supersaturation-driven fabrication of its fiber network. AB - The architecture of a biocompatible organogel formed by gelation of a small molecule organic gelator, N-lauroyl-L-glutamic acid di-n-butylamide, in isostearyl alcohol was investigated based on a supersaturation-driven crystallographic mismatch branching mechanism. By controlling the supersaturation of the system, the correlation length that determines the mesh size of the fiber network was finely tuned and the rheological properties of the gel were engineered. This approach is of considerable significance for many gel-based applications, such as controlled release of drugs that requires precise control of the mesh size. A direct cryo-transmission electron microscopy (TEM) imaging technique capable of preserving the network structure was used to visualize its nanostructure. PMID- 16375419 TI - A site-isolated rhodium-diethylene complex supported on highly dealuminated Y zeolite: synthesis and characterization. AB - The reaction of Rh(C2H4)2(acac) with the partially dehydroxylated surface of dealuminated zeolite Y (calcined at 773 K) and treatments of the resultant surface species in various atmospheres (He, CO, H2, and D2) were investigated with infrared (IR), extended X-ray absorption fine structure (EXAFS), and 13C NMR spectroscopies. The IR spectra show that Rh(C2H4)2(acac) reacted readily with surface OH groups of the zeolite, leading to loss of acac ligands from the Rh(C2H4)2(acac) and formation of supported mononuclear rhodium complexes, confirmed by the lack of Rh-Rh contributions in the EXAFS spectra; each Rh atom was bonded on average to two oxygen atoms of the zeolite surface with a Rh-O distance of 2.19 A. IR, EXAFS, and 13C NMR spectra show that the ethylene ligands remained bonded to the Rh center in the supported complex. Treatment of the sample in CO led to the formation of site-isolated Rh(CO)2 complexes bonded to the zeolite. The sharpness of the nu(CO) bands in the IR spectrum gives evidence of a nearly uniform supported Rh(CO)2 complex and, by inference, the near uniformity of the mononuclear rhodium complex with ethylene ligands from which it was formed. The supported complex with ethylene ligands reacted with H2 to give ethane, and it also catalyzed ethylene hydrogenation at 294 K. PMID- 16375420 TI - Structure of hydrated Na-Nafion polymer membranes. AB - We use molecular dynamics simulations to investigate the structure of the hydrated Na-Nafion membranes. The membrane is "prepared" by starting with the Nafion chains placed on a cylinder having the water inside it. Minimizing the energy of the system leads to a filamentary hydrophilic domain whose structure depends on the degree of hydration. At 5 wt % water the system does not have enough water molecules to solvate all the ions that could be formed by the dissociation of the -SO3Na groups. As a result, the -SO3Na groups aggregate with the water to form very small droplets that do not join into a continuous phase. The size of the droplets is between 5 and 8 A. As the amount of water present in the membrane is increased, the membrane swells, and SO3Na has an increasing tendency to dissociate into ions. Furthermore, a transition to a percolating hydrophilic network is observed. In the percolating structure, the water forms irregular curvilinear channels branching in all directions. The typical dimension of the cross section of these channels is about 10-20 A. Calculated neutron scattering from the simulated system is in qualitative agreement with experiment. In all simulations, the pendant sulfonated perfluorovinyl side chains of the Nafion hug the walls of the hydrophilic channel, while the sulfonate groups point toward the center of the hydrophilic phase. The expulsion of the side chains from the hydrophilic domain is favored because it allows better interaction between the water molecules. We have also examined the probability of finding water molecules around the Na+ and the -SO3(-) ions as well as the probability of finding other water molecules next to a given water molecule. These probabilities are much broader than those found in bulk water or for one ion in bulk water (calculated with the potentials used in the present simulation). This is due to the highly inhomogeneous nature of the material contained in the small hydrophilic pores. PMID- 16375421 TI - Nanocrystalline TiO2/ZnO thin films: fabrication and application to dye sensitized solar cells. AB - Nanocrystalline TiO2 thin films composed of densely packed grains were deposited onto indium-doped tin oxide (ITO)-coated glass substrates at room temperature using a chemical bath deposition technique. A layer-by-layer (LbL) process was utilized to obtain a 1.418-microm-thick TiO2/ZnO structure. The TiO2 surface was super-hydrophilic, but its hydrophilicity decreased considerably after ZnO deposition. Other TiO2/ZnO films were studied to assess their suitability as photoelectrodes in dye-sensitized solar cells (DSSCs). PMID- 16375422 TI - Visible light active platinum-ion-doped TiO2 photocatalyst. AB - Platinum-ion-doped TiO2 (Pt(ion)-TiO2) was synthesized by a sol-gel method, and its visible light photocatalytic activities were successfully demonstrated for the oxidative and reductive degradation of chlorinated organic compounds. Pt(ion) TiO2 exhibited a yellow-brown color, and its band gap was lower than that of undoped TiO2 by about 0.2 eV. The flat band potential of Pt(ion)-TiO2 was positively shifted by 50 mV compared with that of undoped TiO2. X-ray absorption spectroscopy and X-ray photoelectron spectroscopy analyses showed that the Pt ions substituted in the TiO2 lattice were present mainly in the Pt(IV) state with some Pt(II) on the sample surface. Pt(ion)-TiO2 exhibited higher photocatalytic activities than undoped TiO2 under UV irradiation as well. The visible light activity of Pt(ion)-TiO2 was strongly affected by the calcination temperature and the concentration of Pt ion dopant, which were optimal at 673 K and 0.5 atom %, respectively. Under visible irradiation, Pt(ion)-TiO2 degraded dichloroacetate and 4-chlorophenol through an oxidative path and trichloroacetate via a reductive path. The activity of Pt(ion)-TiO2 was not reduced when used repeatedly under visible light. However, visible-light-illuminated Pt(ion)-TiO2 could not degrade substrates such as tetramethylammonium and trichloroethylene, which are degraded with UV-illuminated TiO2. The characteristics and reactivities of Pt(ion)-TiO2 as a new visible light photocatalyst were investigated in various ways and discussed in detail. PMID- 16375423 TI - Stimulated emissions in aligned CdS nanowires at room temperature. AB - Aligned CdS nanowires (NWs) were obtained through a simple thermal evaporation process with highly active CdS nanoparticles as the evaporation source. These NWs show prominent optical waveguides behavior under a continuous-wave (CW) laser excitation. Excitation intensity-dependent photoluminescence (PL) measurements show that these NWs exhibited both broad and supernarrow stimulated emission (lasing) under intense pulse optical excitation at room temperature. Raman scattering and time-resolved PL measurements were used to investigate the optical properties. The results indicated that the stimulated emission in these NWs involves the electron-hole plasma (EHP) and Farby-Perot (F-P) optical resonant processes at room temperature. PMID- 16375424 TI - Combined DFT theoretical calculation and solid-state NMR studies of Al substitution and acid sites in zeolite MCM-22. AB - The distributions of Bronsted acidic protons and their acid strengths in zeolite H-MCM-22 have been characterized by density functional theory (DFT) calculations as well as magic angle spinning (MAS) NMR experiments. The embedded scheme (ONIOM) that combines the quantum mechanical (QM) description of active sites and semiempirical AM1 treatment of the neighboring environment was applied to predict the aluminum substitution mechanism and proton affinity (PA), as well as adsorption behaviors of acetone and trimethylphosphine oxide (TMPO) onto the zeolite. Our theoretical results indicate that the Al substitution takes place in the order of Al1-OH-Si2 > Al8-OH-Si8 > Al5-OH-Si7. The DFT theoretical and NMR experimental results suggest that the acid strength of the three Bronsted acid sites in H-MCM-22 zeolite is slightly lower than that of H-ZSM-5 zeolite and the accessible Bronsted acidic protons most likely reside in both the supercages (at the Al8-OH-Si8 and Al1-OH-Si2 sites) and external surface pocket (at the Al8-OH Si8 site) rather than in the sinusoidal channels (Al5-OH-Si7), with the Al1-OH Si2 site having the strongest acid strength (as probed by TMPO). This may partially explain the special selectivity of acid-catalyzed reactions occurring inside the channels of H-MCM-22 zeolite. PMID- 16375425 TI - Molecular dynamics simulation of titanium dioxide nanoparticle sintering. AB - Nanoparticles have been an area of active research in recent years due to their properties, which can be greatly different from the bulk. In this work, we study the sintering of TiO2 nanoparticles using molecular dynamics simulations. Such sintering occurs in flame reactors where nanotitania is prepared via the chloride process. Decrease in free energy due to reduction in surface area is the main driving force for sintering of particles. Simulations, at various starting temperatures and orientations, indicate that the process of sintering is strongly affected by temperature and initial orientation. Extremely high diffusion of ions in the neck region of sintering nanoparticles supports the idea that solid-state diffusion is significant in metal-oxide nanoparticle sintering. It is found that the dipole-dipole interaction between sintering nanoparticles plays a very important role at temperatures away from the melting point. The duration of the simulation is not enough to observe the complete sintering process, but important initial stages are well studied. PMID- 16375426 TI - Self-assembly of gold nanoparticles induced by poly(oxypropylene)diamines. AB - A series of poly(oxypropylene)diamines D230, D400, D2000, and D4000, having molecular weights of hydrophobic segments of 230, 400, 2000, and 4000, were used as ligands to synthesize self-organized gold nanocrystals. Ligand exchange significantly reduced the average particle size and the polydispersity of nanocrystals, and this effect was more remarkable as the molar ratio of amine groups to Au3+ ions ([N]/[Au3+] ratio) was increased. Under the same [N]/[Au3+] ratio of 100, D2000 generated an ordered 2D-monolayer; however, D230 and D400 colloids formed mainly a densely packed 3D structure with minor 2-D layers, and D4000 presented disordered 3D and 2D structures. The gap among the nanoparticles was found to be increased with the increasing molecular weight of the hydrophobic segment of ligands, accompanied by the decreasing wavelength of UV-vis absorption bands. This increased gap can be interpreted as the ligand thickness calculated from the equation of steric force increasing with increasing molecular weight of the hydrophobic segment. The potential energies obtained from the calculated ligand thickness according to the soft sphere model show more steep potential wells for D230 and D400 colloids than that for the D2000 colloid. This explains why the aggregation hardly occurred for the gold nanoparticles obtained under D2000, where the nanoparticles are single crystals having face center crystal structure with a lattice constant of 2.36 A and have grain sizes close to the average particle sizes, evidenced from the results of transmission electron microscopy and X-ray diffraction spectroscopy. PMID- 16375427 TI - Pore morphology of porous polymer particles probed by NMR relaxometry and NMR cryoporometry. AB - The pore size distribution (PSD) and pore connectivity (PC) within porous polymer particles are probed by combining NMR cryoporometry and NMR relaxometry (spin spin relaxation). With water as a probe molecule, the constant K in the so-called Gibbs-Thompson equation and the surface relaxivity (rho2) were determined to be K = (420 +/- 50) KA and rho2 = (0.44 +/- 0.01) x 10(-6) ms(-1), respectively. Also, the thickness of the interface layer was estimated to be of the order of one monolayer of water molecules. A detailed analysis of the complete set of NMR data enabled the morphology or pore structure to be probed, and is thoroughly discussed in the text. PMID- 16375428 TI - Comparison of all sites for Ti substitution in zeolite TS-1 by an accurate embedded-cluster method. AB - We studied the preferential location of Ti centers in the framework of the Ti containing MFI zeolite TS-1 using a hybrid DFT/MM embedding method developed recently. This "covalent elastic polarizable environment" (covEPE) cluster embedding allows a complete and self-consistent treatment of solid covalent systems such as zeolites. For the present study, we used a gradient-corrected density functional approach. The resulting structural features of both Si- and Ti substituted forms of the zeolite framework fit well with available experimental information. The calculated substitution energy of Ti at the 12 crystallographically different tetrahedral sites of the MFI structure vary within 19 kJ/mol with T12 and T2 as most and least preferred sites, respectively. On the basis of these computational results and the preferential sites for Ti substitution reported from different experimental investigations, we concluded that the Ti distribution in the TS-1 framework is not governed by the thermodynamic stability of the pure material. PMID- 16375429 TI - Study of microstructural characterization and ionic conductivity of a chemical covalent polyether-siloxane hybrid doped with LiClO4. AB - The chemical-covalent polyether-siloxane hybrids (EDS) doped with various amounts of LiClO4 salt were characterized by FT-IR, DSC, TGA, and solid-state NMR spectra as well as impedance measurements. These observations indicate that different types of complexes by the interactions of Li+ and ClO4- ions are formed within the hybrid host, and the formation of transient cross-links between Li+ ions and ether oxygens results in the increase in T(g) of polyether segments and the decrease in thermal stability of hybrid electrolyte. Initially a cation complexation dominated by the oxirane-cleaved cross-link site and PEO block is present, and after the salt-doped level of O/Li+ = 20, the complexation through the PPO block becomes more prominent. Moreover, a significant degree of ionic association is examined in the polymer-salt complexes at higher salt uptakes. A VTF-like temperature dependence of ionic conductivity is observed in all of the investigated salt concentrations, implying that the diffusion of charge carrier is assisted by the segmental motions of the polymer chains. The behavior of ion transport in these hybrid electrolytes is further correlated with the interactions between ions and polymer host. PMID- 16375430 TI - Synthesis and characterization of organofunctionalized MCM-41 by the original stepped templated sol-gel technology. AB - Four types of organofunctionalized (by phenyl, 3-aminopropyl, 3-chloropropyl, and N-aminoethylaminopropyl, respectively) mesoporous silicates MCM-41 (hereafter R MCM-41) have been successfully synthesized via the grafting method, the conventional templated sol-gel technology, and the original stepped templated sol gel technology. The prepared inorganic-organic hybrid materials were characterized in detail by powder X-ray diffraction (XRD), 29Si and 13C MAS NMR spectrometry, isothermal nitrogen adsorption-desorption analysis, FT-IR spectrometry including in situ drift technique, and thermogravimetric analysis (TGA). Through a series of investigations and comparisons, the advantageous stepped templated sol-gel technology introduced in this paper is demonstrated to be a remarkable approach that gathers the superior properties of the current grafting method and the conventional hydrothermal co-condensation synthesis, yielding as a consequence both thermodynamically firm fixation and relatively large content of organic functional species on the MCM-41 support. The novel synthetic strategy also evidently shortens the crystallization time for the organofunctionalized MCM-41 with regular specific structure. PMID- 16375431 TI - Size-dependent orientation growth of large-area ordered Ni nanowire arrays. AB - Large-area ordered Ni nanowire arrays with different diameters have been fabricated by the direct current electrodeposition into the holes of porous anodic alumina membrane. The crystal structure and micrograph of nanowire arrays are characterized by X-ray diffraction, field-emission scanning electron microscopy and high-resolution transmission electron microscopy. The results indicate that the growth orientation of Ni nanowires turns from [110] to [111] direction with increasing diameters of nanowires. The mechanism of the growth was discussed in terms of interface energy minimum principle. The size-dependent orientation of Ni nanowire arrays has the important significance for the design and control of nanostructures. PMID- 16375432 TI - Synthesis, structure, and reactivity of a new Ti-containing microporous/mesoporous material. AB - A new class of porous, mixed phase titanosilicate materials containing a microporous TS-1 phase and a mesoporous Ti-MCM-48 phase has been successfully synthesized. A novel, one-pot synthesis method was used in which the organic templates for the mesoporous and microporous phases were added sequentially to the same reaction mixture, followed by crystallization at 150 degrees C. The gemini surfactant 18-12-18 was used to form the Ti-MCM-48 mesophase; subsequent addition of tetrapropylammonium cation (TPA+) led to the formation of TS-1. The relative amounts of the two phases within the final products were controlled by optimizing the crystallization time. Crystallization times between 12 and 50 h gave materials containing both phases, with an increasing amount of microphase formed at longer crystallization times. These materials, called "Ti-MMM-2" (microporous/mesoporous materials) were characterized using powder XRD, N2 physisorption, TEM, FTIR, DR-UV/Vis spectroscopy, and 29Si MAS NMR. In the epoxidation of cyclohexene with tert-butyl hydroperoxide (TBHP), Ti-MMM-2 samples exhibited higher catalytic activity (approximately 61%) than either TS-1 (16%) or Ti-MCM-48 (42%), with a very high selectivity (97%) for formation of cyclohexene oxide. PMID- 16375433 TI - Dielectric studies of tetraethylene glycol-bis(3-methylimidazolium) dichloride (TEGDC) exhibiting large negative dielectric anisotropy. AB - A dielectric anisotropy property of a TEGDC (tetraethylene glycol-bis(3 methylimidazolium) dichloride) is investigated as a function of frequency. TEGDC showed an extremely large negative dielectric anisotropy (-10.95 to -4753.73). Variation of dielectric anisotropy (delta epsilon) with respect to the spot frequencies reveals that liquid crystal (LC) orientation has an n-type property at low frequencies and as the frequency increases dielectric anisotropy character shifts from negative dielectric anisotropy type (n-type) to positive dielectric anisotropy type (p-type). Consequently, the TEGDC is a liquid crystal with large negative dielectric anisotropy. PMID- 16375434 TI - Determination of pore sizes and volumes of porous materials by 129Xe NMR of xenon gas dissolved in a medium. AB - In our previous paper (J. Phys. Chem. B 2005, 109, 757) it was illustrated that the 129Xe NMR spectra of xenon dissolved in acetonitrile confined into mesoporous materials give detailed information on the system, especially about the pore sizes. A resonance signal originating from xenon atoms sited in very small cavities built up inside the pores during the freezing transition (referred to as signal D) turned out to be highly sensitive to the pore size. The emergence of this signal reveals the phase transition temperature of acetonitrile inside the pores, which can also be used to determine the size of the pores. In addition, the difference in the chemical shifts of two other signals arising from xenon dissolved in bulk and confined acetonitrile (B and C) provides another method for determining the pore sizes. In the present work, the observed correlations have been investigated using an extensive set of measurements with a variety of porous materials (silica gels and controlled pore glasses) with the mean pore diameters ranging from 43 to 2917 A. The usefulness of the correlations has been demonstrated by calculating the pore size distributions from the spectral data. The distributions are in agreement with those reported by the manufacturers, when the mean pore diameter is smaller than approximately 500 A. In addition, it has been shown that the porosity of the materials can be determined by comparing the intensities of the signals arising from the bulk and confined liquid. When acetonitrile is replaced by cyclohexane in the sample, the dependence of the chemical shift difference between the B and C signals on the pore size becomes more sensitive, but no D signal appears below the freezing point. In addition, the influence of xenon gas on the melting points of bulk and confined acetonitrile has been studied by 1H NMR cryoporometry. The measurements show that the temperature of the latter transition lowers slightly more, and consequently affects the pore sizes calculated by means of the difference in the phase transition temperatures. Hysteresis in the phase transitions in a cooling-warming cycle has also been studied as a function of the temperature stabilization time by 129Xe NMR of xenon dissolved in acetonitrile. PMID- 16375435 TI - Evolution of the properties of Al(n)N(n) clusters with size. AB - A global optimization of stoichiometric (AlN)(n) clusters (n = 1-25, 30, 35, ..., 95, 100) has been performed using the basin-hopping (BH) method and describing the interactions with simple and yet realistic interatomic potentials. The results for the smaller isomers agree with those of previous electronic structure calculations, thus validating the present scheme. The lowest-energy isomers found can be classified in three different categories according to their structural motifs: (i) small clusters (n = 2-5), with planar ring structures and 2-fold coordination, (ii) medium clusters (n = 6-40), where a competition between stacked rings and globular-like empty cages exists, and (iii) large clusters (n > 40), large enough to mix different elements of the previous stage. All the atoms in small and medium-sized clusters are in the surface, while large clusters start to display interior atoms. Large clusters display a competition between tetrahedral and octahedral-like features: the former lead to a lower energy interior in the cluster, while the latter allow for surface terminations with a lower energy. All of the properties studied present different regimes according to the above classification. It is of particular interest that the local properties of the interior atoms do converge to the bulk limit. The isomers with n = 6 and 12 are specially stable with respect to the gain or loss of AlN molecules. PMID- 16375436 TI - Controlled synthesis, characterization, and crystallization of Ni-P nanospheres. AB - The size- and composition-controlled synthesis of Ni-P nanospheres from nickel chloride and sodium hypophosphite has been systematically investigated by changing the conditions, such as the ratio of the starting materials, pH value, and reduction temperature. It was found that when the starting ratio of H2PO2( )/Ni2+ was changed the size and chemical composition of the nanoparticles changed simultaneously. Within a suitable pH range, the phosphorus content was altered without affecting the particle size. Increasing the reduction temperature resulted in smaller Ni-P nanospheres but invariable phosphorus content. The Ni-P nanospheres were amorphous when the phosphorus content was higher than 10.0 mol %, while lower phosphorus content led to a composite of amorphous Ni-P and face centered cubic (fcc) Ni. During postsynthesis calcinations, amorphous Ni-P nanospheres with a low phosphorus content directly crystallized to Ni3P and fcc Ni. However, the specimens with high phosphorus content crystallized via some intermediate phases such as Ni5P2 and Ni12P5. In the latter, an amorphous P-rich shell was developed simultaneously. A preliminary catalytic test of growth of carbon nanofibers on the Ni-P nanospheres has been carried out. PMID- 16375437 TI - Electroviscous effect of moderately concentrated colloidal suspensions: Stern layer influence. AB - A previous model for the viscosity of moderately concentrated suspensions has been extended. The influence of a dynamic Stern layer (DSL), which produces an additional surface conductance at the electrolyte-particle interface, is included. The theoretical treatment is based on Happel's cell model with Simha's boundary conditions for the interparticle hydrodynamic interactions and on a dynamic Stern-layer model for ionic conduction on the particle surface according to Mangelsdorf and White (ref 39). The results are valid for arbitrary zeta potentials and double-layer thickness. Extensive theoretical predictions are shown and interesting new behaviors are found. The comparison with the results in the absence of additional surface conductance shows a great influence of this mechanism in the energy dissipation during the laminar flow of these suspensions. We conclude that the inclusion of a dynamic Stern layer will be required to match the predictions with the experimental results. PMID- 16375438 TI - Shape-selective synthesis and oxygen storage behavior of ceria nanopolyhedra, nanorods, and nanocubes. AB - Single-crystalline and uniform nanopolyhedra, nanorods, and nanocubes of cubic CeO2 were selectively prepared by a hydrothermal method at temperatures in the range of 100-180 degrees C under different NaOH concentrations, using Ce(NO3)3 as the cerium source. According to high-resolution transmission electron microscopy, they have different exposed crystal planes: {111} and {100} for polyhedra, {110} and {100} for rods, and {100} for cubes. During the synthesis, the formation of hexagonal Ce(OH)3 intermediate species and their transformation into CeO2 at elevated temperature, together with the base concentration, have been demonstrated as the key factors responsible for the shape evolution. Oxygen storage capacity (OSC) measurements at 400 degrees C revealed that the oxygen storage takes place both at the surface and in the bulk for the as-obtained CeO2 nanorods and nanocubes, but is restricted at the surface for the nanopolyhedra just like the bulk one, because the {100}/{110}-dominated surface structures are more reactive for CO oxidation than the {111}-dominated one. This result suggests that high OSC materials might be designed and obtained by shape-selective synthetic strategy. PMID- 16375439 TI - Second harmonic generation phase measurements of Cr(VI) at a buried interface. AB - Surface second harmonic generation (SHG) phase measurements are carried out on methyl ester-functionalized fused quartz/water interfaces in the presence and absence of Cr(VI). The experiments are performed at pH 7, room temperature, and a chromate concentration of 10(-4) M, which corresponds to monolayer Cr(VI) coverage. The liquid/solid interface is probed from the fused quartz side by directing the probe light field at 580 nm onto the interface together with an SHG reference signal at 290 nm that is collinear with the fundamental. The phase difference of the SHG signals generated at the interface in the presence and absence of Cr(VI) is 85 degrees, which is consistent with SHG resonance enhancement observed for the surface-bound Cr(VI) near 290 nm. The optical arrangement discussed here does not require vacuum technology or optics that compensate for the dispersion of the fundamental and the second harmonic E-fields in the two condensed-phase media. This approach is general and can be applied for analyzing thermodynamic and kinetic data derived from SHG measurements of physical and chemical processes occurring at any buried interface. PMID- 16375440 TI - Isotope fractionation of water during evaporation without condensation. AB - The microscopic events engendering liquid water evaporation have received much attention over the last century, but remain incompletely understood. We present measurements of isotope fractionation occurring during free molecular evaporation from liquid microjets and show that the isotope ratios of evaporating molecules exhibit dramatic differences from equilibrium vapor values, strong variations with the solution deuterium mole fraction, and a clear temperature dependence. These results indicate the existence of an energetic barrier to evaporation and that the evaporation coefficient of water is less than unity. These new insights into water evaporation promise to advance our understanding of the processes that control the formation and lifetime of clouds in the atmosphere. PMID- 16375441 TI - Electrochemical derivatization of carbon surface by reduction of in situ generated diazonium cations. AB - The derivatization of a glassy carbon electrode surface was achieved by electrochemical reduction of several in situ generated diazonium cations. The diazonium cations were synthesized in the electrochemical cell by reaction of the corresponding amines with NaNO2 in aqueous HCl. The versatility of the method was demonstrated by using six diazonium cations. This deposition method, which involves simple reagents and does not require the isolation and purification of the diazonium salt, enabled the grafting of covalently bounded layers which exhibited properties very similar to those of layers obtained by the classical derivatization method involving isolated diazonium salt dissolved in acetonitrile or aqueous acid solution. Cyclic voltammetry and electrochemical impedance spectroscopy carried out in aqueous solutions containing electroactive redox probe molecules such as Fe(CN)6(3-/4-) and Ru(NH3)6(3+) confirmed the barrier properties of the deposited layers. The chemical composition of the grafted layers was determined by X-ray photoelectron spectroscopy and surface coverage in the range 3 x 10(-10) to 6 x 10(-10) mol cm(-2) was estimated for films grown in our experimental conditions. PMID- 16375442 TI - Ultrathin TiO(x) films on Pt(111): a LEED, XPS, and STM investigation. AB - Ultrathin ordered titanium oxide films on Pt(111) surface are prepared by reactive evaporation of Ti in oxygen. By varying the Ti dose and the annealing conditions (i.e., temperature and oxygen pressure), six different long-range ordered phases are obtained. They are characterized by means of low-energy electron diffraction (LEED), X-ray photoemission spectroscopy (XPS), and scanning tunneling microscopy (STM). By careful optimization of the preparative parameters, we find conditions where predominantly single phases of TiO(x), revealing distinct LEED pattern and STM images, are produced. XPS binding energy and photoelectron diffraction (XPD) data indicate that all the phases, except one (the stoichiometric rect-TiO2), are one monolayer thick and composed of a Ti-O bilayer with interfacial Ti. Atomically resolved STM images confirm that these TiO(x) phases wet the Pt surface, in contrast to rect-TiO2. This indicates their interface stabilization. At a low Ti dose (0.4 monolayer equivalents, MLE), an incommensurate kagome-like low-density phase (k-TiO(x) phase) is observed where hexagons are sharing their vertexes. At a higher Ti dose (0.8 MLE), two denser phases are found, both characterized by a zigzag motif (z- and z'-TiO(x) phases), but with distinct rectangular unit cells. Among them, z'-TiO(x), which is obtained by annealing in ultrahigh vacuum (UHV), shows a larger unit cell. When the postannealing of the 0.8 MLE deposit is carried out at high temperatures and high oxygen partial pressures, the incommensurate nonwetting, fully oxidized rect TiO2 is found The symmetry and lattice dimensions are almost identical with rect VO2, observed in the system VO(x)/Pd(111). At a higher coverage (1.2 MLE), two commensurate hexagonal phases are formed, namely the w- [(square root(43) x square root(43)) R 7.6 degrees] and w'-TiO(x) phase [(7 x 7) R 21.8 degrees]. They show wagon-wheel-like structures and have slightly different lattice dimensions. Larger Ti deposits produce TiO2 nanoclusters on top of the different monolayer films, as supported both by XPS and STM data. Besides the formation of TiO(x) surfaces phases, wormlike features are found on the bare parts of the substrate by STM. We suggest that these structures, probably multilayer disordered TiO2, represent growth precursors of the ordered phases. Our results on the different nanostructures are compared with literature data on similar systems, e.g., VO(x)/Pd(111), VO(x)/Rh(111), TiO(x)/Pd(111), TiO(x)/Pt(111), and TiO(x)/Ru(0001). Similar and distinct features are observed in the TiO(x)/Pt(111) case, which may be related to the different chemical natures of the overlayer and of the substrate. PMID- 16375443 TI - Computer simulations of catanionic surfactants adsorbed at air/water interfaces. AB - Structural properties pertaining to the solvation of mixtures of dodecytrimethylammonium/dodecylsulfate adsorbed at water/air interfaces were studied using molecular dynamics techniques. Two different surfactant coverages, both in the submonolayer regime, were considered: an infinite-diluted catanionic pair and an equimolar mixture, at a surface concentration of 78.7 A2/headgroup. The most stable solvated structures for the single surfactant pair correspond to contact-head-ion-pairs (CHIP) at a distance close to 5 A. In addition, marginally stable solvent-separated-head-ion-pairs (SSHIP) at distances approximately 7 A were also observed. The mean free energy for the dissociation of CHIP was estimated to be approximately 1 kcal/mol. At finite surfactant concentrations, one observes a considerable degree of clustering between the amphiphiles, due to the strong Coulomb coupling between headgroups. The resulting spatial domains show asymmetric structures with linear dimensions comparable to the simulation box, suggesting the onset of percolative structures. The connectivity pattern of these domains was interpreted in terms of a simplified model consisting of two dimensional charged Lennard-Jones spheres. PMID- 16375444 TI - Intercalation mechanism and interlayer structure of hexadecylamines in the confined space of layered alpha-zirconium phosphates. AB - Well-defined hexadecylamine (HDA) intercalated structures, either interdigitated layers, bilayers, or hybrid layers of both, in a confined space of highly functionalized layered alpha-zirconium phosphates (alpha-ZrPs) have been prepared based on the two-step intercalation mechanism and these distinct intercalated structures can serve as a model system to investigate the interactions of two monolayers whose amphiphilic tails are adjacent to each other. At the first intercalation step, the electrostatic interaction between HDAs and alpha-ZrP is dominant and results in an interdigitated layer structure (d(001) = 3.0 nm) and the interdigitated layer is saturated at around phi = 50%, where phi is the weight fraction of intercalated HDAs in the intergallery of alpha-ZrP. For phi higher than 50%, the bilayer structure (d(001) = 4.3 nm) emerges due to further hydrophobic interaction between HDAs initially grafted to alpha-ZrP and unanchored HDAs and the relative fraction of the bilayer structure over the interdigitated layer increases with the increase in the intercalated amount of HDAs. The intriguing morphology of alpha-ZrP tactoids intercalated with HDAs in coexisting bilayers and interdigitated layers is observed by using microtomed TEM and the two-step intercalation has also been verified with TGA and FT-IR. Also, a structural transition from the bilayers to the interdigitated layers is monitored by using in situ synchrotron WAXS showing that the hydrophobically intercalated HDAs are selectively deintercalated at a relatively low decomposition temperature around 220 degrees C. PMID- 16375445 TI - Photoelectrochemical behavior of Nb-doped TiO2 electrodes. AB - The photoelectrochemical behavior of degenerate Nb-doped TiO2 (Ti(1-x)Nb(x)O2: x = 0, 0.01, 0.03, 0.06, 0.1) electrodes prepared by pulsed laser deposition on LaAlO3 (LAO) and SrTiO3 (STO) was examined, revealing that an increase in Nb concentration causes a significant decay of titania photoactivity. One reason for such behavior may be a Burstein-Moss effect, which leads to a blue shift of the spectral limit of photoactivity. Another reason typical for metal-doped photocatalysts is the increase of the efficiency of charge carrier recombination. PMID- 16375446 TI - Study of adsorption and decomposition of H2O on Ge(100). AB - The adsorption and decomposition of water on Ge(100) have been investigated using real-time scanning tunneling microscopy (STM) and density-functional theory (DFT) calculations. The STM results revealed two distinct adsorption features of H2O on Ge(100) corresponding to molecular adsorption and H-OH dissociative adsorption. In the molecular adsorption geometry, H2O molecules are bound to the surface via Ge-O dative bonds between the O atom of H2O and the electrophilic down atom of the Ge dimer. In the dissociative adsorption geometry, the H2O molecule dissociates into H and OH, which bind covalently to a Ge-Ge dimer on Ge(100) in an H-Ge-Ge-OH configuration. The DFT calculations showed that the dissociative adsorption geometry is more stable than the molecular adsorption geometry. This finding is consistent with the STM results, which showed that the dissociative product becomes dominant as the H2O coverage is increased. The simulated STM images agreed very well with the experimental images. In the real-time STM experiments, we also observed a structural transformation of the H2O molecule from the molecular adsorption to the dissociative adsorption geometry. PMID- 16375447 TI - Surface modification and laser pulse length effects on internal energy transfer in DIOS. AB - Benzyl-substituted benzylpyridinium (BP) chloride salts were used as a source of thermometer ions to probe the internal energy (IE) transfer in desorption/ionization on porous silicon (DIOS). To modify their wetting properties and the interaction energies with the thermometer ions, the DIOS surfaces were silylated to produce trimethylsilyl- (TMS), amine- (NH2), perfluoroalkyl- (PFA), and perfluorophenyl-derivatized (PFP) surfaces. Two laser sources--a nitrogen laser with pulse length of 4 ns and a mode locked 3 x omega Nd:YAG laser with a pulse length of 22 ps--were utilized to induce desorption/ionization and fragmentation at various laser fluence levels. The corresponding survival yields were determined as indicators of the IE transfer and the IE distributions were extracted. In most cases, with increasing the laser fluence in a broad range (approximately 20 mJ/cm2), no change in IE transfer was observed. For ns excitation, this was in remarkable contrast with MALDI, where increasing the laser fluence resulted in sharply (within approximately 5 mJ/cm2) declining survival yields. Derivatization of the porous silicon surface did not affect the survival yields significantly but had a discernible effect on the threshold fluence for ion production. The IE distributions determined for DIOS and MALDI from alpha-cyano-4-hydroxycinnamic acid reveal that the mean IE value is always lower for the latter. Using the ps laser, the IE distribution is always narrower for DIOS, whereas for ns laser excitation the width depends on surface modification. Most of the differences between MALDI and DIOS described here are compatible with the different dimensionality of the plume expansion and the differences in the activation energy of desorption due to surface modifications. PMID- 16375448 TI - Enzyme-like kinetics of ferryloxy myoglobin formation in films on electrodes in microemulsions. AB - Covalently linked films of the ferric heme protein myoglobin and poly-L-lysine on pyrolytic graphite electrodes reacted with tert-butylhydroperoxide (tBuOOH) to form ferryloxy protein species according to Michaelis-Menten enzyme kinetics. Rotating disk voltammetry data obtained in microemulsions, micellar solution, and buffers revealed a strong influence of water phase acidity on kinetic parameters. Microemulsion and surfactant type had a much smaller influence on reaction kinetics, possibly because the reaction takes place entirely in a water environment surrounding Mb in the films in all fluids. A large apparent Michaelis kcat in microemulsions with neutral water phases was offset by much weaker binding as shown by larger protein-substrate dissociation constants (Km). Acidic SDS microemulsions and pH 2 buffer provided the most efficient reaction conditions as judged by the ratio kcat/Km. Apparent kinetic constants are most likely governed by acidity-controlled protein conformations and their binding with tBuOOH in the intermediate protein-substrate complex. PMID- 16375449 TI - Equilibrium morphology of face-centered cubic gold nanoparticles >3 nm and the shape changes induced by temperature. AB - Many of the unique properties of metallic nanoparticles are determined not only by their finite size but also by their shape, defined by the crystallographic orientation of the surface facets. These surfaces (and therefore the nanoparticles themselves) may differ in a number of ways, including surface atom densities, electronic structure, bonding, chemical reactivities, and thermodynamic properties. In the case of gold, it is known that the melting temperature of nanoparticles strongly depends on the crystal size and that the shape may alter considerably (and yet somewhat unpredictably) during annealing. In this work we use first principle calculations and a thermodynamic model to investigate the morphology of gold nanoparticles in the range 3-100 nm. The results predict that the equilibrium shape of gold nanoparticles is a modified truncated octahedron and that the (size-dependent) melting of such particles is preceded by a significant change in the nanoparticle's morphology. PMID- 16375450 TI - A molecular dynamics study of the thermodynamic properties of calcium apatites. 1. Hexagonal phases. AB - Structural and thermodynamic properties of crystal hexagonal calcium apatites, Ca10(PO4)6(X)2 (X = OH, F, Cl, Br), were investigated using an all-atom Born Huggins-Mayer potential by a molecular dynamics technique. The accuracy of the model at room temperature and atmospheric pressure was checked against crystal structural data, with maximum deviations of ca. 4% for the haloapatites and 8% for hydroxyapatite. The standard molar lattice enthalpy, delta(lat)H298(o), of the apatites was calculated and compared with previously published experimental results, the agreement being better than 2%. The molar heat capacity at constant pressure, C(p,m), in the range 298-1298 K, was estimated from the plot of the molar enthalpy of the crystal as a function of temperature, H(m) = (H(m,298) - 298C(p,m)) + C(p,m)T, yielding C(p,m) = 694 +/- 68 J x mol(-1) x K(-1), C(p,m) = 646 +/- 26 J x mol(-1) x K(-1), C(p,m) = 530 +/- 34 J x mol(-1) x K(-1), and C(p,m) = 811 +/- 42 J x mol(-1) x K(-1) for hydroxy-, fluor-, chlor-, and bromapatite, respectively. High-pressure simulation runs, in the range 0.5-75 kbar, were performed in order to estimate the isothermal compressibility coefficient, kappaT, of those compounds. The deformation of the compressed solids is always elastically anisotropic, with BrAp exhibiting a markedly different behavior from those displayed by HOAp and ClAp. High-pressure p-V data were fitted to the Parsafar-Mason equation of state with an accuracy better than 1%. PMID- 16375451 TI - Orientational dynamics in the isotropic phase of a calamitic liquid-crystal model. AB - We report a molecular dynamics simulation study on the isotropic phase of an idealized calamitic liquid crystal model with a length-to-width ratio of approximately 5-6. The study focuses on the characterization of single-particle and collective orientational dynamics on approaching the phase transition to the nematic phase. Recent experimental and simulation works have suggested that a power law behavior exists at relatively short times in the decay of the time derivative of the orientational correlation functions. Qualitatively, our simulation data are consistent with these findings. Both single-particle and collective time correlation function derivatives possess, in their respective log log plots, a linear region at very short times, whose slope is essentially independent from the thermodynamic state. Nevertheless, the single-particle orientational correlation functions are better described by a function which is the sum of a fast exponential, an intermediate stretched-exponential and a slow exponential, while the collective orientational correlation functions are satisfactorily described by a sum of two exponentials, at higher density, or by just one exponential, at lower density. PMID- 16375452 TI - Specific ion effects in solutions of globular proteins: comparison between analytical models and simulation. AB - Monte Carlo simulations have been performed for ion distributions outside a single globular macroion and for a pair of macroions, in different salt solutions. The model that we use includes both electrostatic and van der Waals interactions between ions and between ions and macroions. Simulation results are compared with the predictions of the Ornstein-Zernike equation with the hypernetted chain closure approximation and the nonlinear Poisson-Boltzmann equation, both augmented by pertinent van der Waals terms. Ion distributions from analytical approximations are generally very close to the simulation results. This demonstrates that properties that are related to ion distributions in the double layer outside a single interface can to a good approximation be obtained from the Poisson-Boltzmann equation. We also present simulation and integral equation results for the mean force between two globular macroions (with properties corresponding to those of hen-egg-white lysozyme protein at pH 4.3) in different salt solutions. The mean force and potential of mean force between the macroions become more attractive upon increasing the polarizability of the counterions (anions), in qualitative agreement with experiments. We finally show that the deduced second virial coefficients agree quite well with experimental results. PMID- 16375453 TI - Surface activity of lysozyme and dipalmitoyl phosphatidylcholine vesicles at compressed and supercritical fluid interfaces. AB - The surface activities of lysozyme and dipalmitoyl phosphatidylcholine (DPPC) vesicles at aqueous/compressed fluid interfaces are examined via high-pressure interfacial tension measurements using the pendant drop technique. The density and interfacial tension in compressible fluid systems vary significantly with pressure, providing a versatile medium for elucidating interactions between biomolecules and fluid interfaces and a method to elicit pressure-dependent interfacial morphological responses. The effects of lysozyme concentration (0.0008, 0.01, and 1 mg/mL) and pressure (> or = 7 MPa) on the dynamic surface response in the presence of ethane, propane, N2, and CO2 at 298 K were examined. Interfacial lysozyme adsorption reduced the induction phase and quickly led to interfacial tensions consistent with protein conformational changes and monolayer saturation at the compressed fluid interfaces. Protein adsorption, as indicated by surface pressure, correlated with calculated Hamaker constants for the compressed gases, denoting the importance of dispersion interactions. For DPPC at aqueous/compressed or aqueous/supercritical CO2 interfaces (1.8-20.7 MPa, 308 K), 2-3-fold reductions in interfacial tension were observed relative to the pure binary fluid system. The resulting surface pressures infer pressure-dependent morphological changes within the DPPC monolayer. PMID- 16375454 TI - Theoretical studies of manganese and iron superoxide dismutases: superoxide binding and superoxide oxidation. AB - Density-functional calculations indicate that the second sphere of coordination around the metal centers of manganese and iron superoxide dismutases (MnSODs and FeSODs) plays an important role in the binding of O2(-). In these systems, O2(-) prefers to bind to Mn or Fe in end-on configurations. For human and E. coli MnSODs, the bound O2(-) forms hydrogen bonds with the tyrosine and glutamine amino acid residues in the second sphere of coordination. In the cases of E. coli and T. elongates FeSODs, hydrogen bonding occurs between the bound O2(-) and the tyrosine amino acid only because the glutamine is too far away for an effective bonding interaction. The manner in which the O2(-) binds to the metal center in MnSODs and FeSODs can affect the rate of subsequent protonation and determine the mechanism for the formation of H2O2. Both Mn- and Fe-containing superoxide dismutases contain a metal-bound solvent molecule that has been suggested to be involved in the uptake of a H+ upon reduction of the metal center [Bull, C.; Fee, J. A. J. Am. Chem. Soc. 1985, 107, 3295; Miller, A.-F.; Padmakumar, K.; Sorkin, D. L.; Karapetian, A.; Vance, C. K. J. Inorg. Biochem. 2003, 93, 71]. Using density-functional theory, we confirm this suggestion and show the involvement of the second sphere of coordination in the process. We show that the oxidation of superoxide by Mn- or Fe-containing superoxide dismutases is facilitated by a cooperative effect between superoxide binding, protonation of the OH- bound to the metal, and electron transfer from the superoxide molecule to the oxidized metal. In particular, proton transfer through tyrosine-34 on the absence of a bound superoxide is uphill while, once superoxide is bound, the energetic barrier is lowered. It is this barrier that likely keeps the resting state (Mn(III)SOD) of the enzyme with a bound hydroxide, instead of a water. This work provides a model for the mechanism of reaction of superoxide with the oxidized form of the metal within Mn- and FeSODs. PMID- 16375455 TI - Time-resolved two-photon spectroscopy of photosystem I determines hidden carotenoid dark-state dynamics. AB - We present time-resolved fs two-photon pump-probe data measured with photosystem I (PS I) of Thermosynechococcus elongatus. Two-photon excitation (lambda(exc)/2 = 575 nm) in the spectral region of the optically forbidden first excited singlet state of the carotenoids, Car S1, gives rise to a 800 fs and a 9 ps decay component of the Car S1 --> S(n) excited-state absorption with an amplitude of about 47 +/- 16% and 53 +/- 10%, respectively. By measuring a solution of pure beta-carotene under exactly the same conditions, only a 9 ps decay component can be observed. Exciting PS I at exactly the same spectral region via one-photon excitation (lambda(exc) = 575 nm) also does not show any sub-ps component. We ascribe the observed constant of 800 fs to a portion of about 47 +/- 16% beta carotene states that can potentially transfer their energy efficiently to chlorophyll pigments via the optically dark Car S1 state. We compared these data with conventional one-photon pump-probe data, exciting the optically allowed second excited state, Car S2. This comparison demonstrates that the fast dynamics of the optically forbidden state can hardly be unravelled via conventional one photon excitation only because the corresponding Car S1 populations are too small after Car S2 --> Car S1 internal conversion. A direct comparison of the amplitudes of the Car S1 --> S(n) excited-state absorption of PS I and beta carotene observed after Car S2 excitation allows determination of a quantum yield for the Car S1 formation in PS I of 44 +/- 5%. In conclusion, an overall Car S2 - > Chl energy-transfer efficiency of approximately 69 +/- 5% is observed at room temperature with 56 +/- 5% being transferred via Car S2 and probably very hot Car S1 states and 13 +/- 5% being transferred via hot and "cold" Car S1 states. PMID- 16375456 TI - Probing the sequence of conformationally induced polarity changes in the molecular chaperonin GroEL with fluorescence spectroscopy. AB - Hydrophobic interactions play a major role in binding non-native substrate proteins in the central cavity of the bacterial chaperonin GroEL. The sequence of local conformational changes by which GroEL and its cofactor GroES assist protein folding can be explored using the polarity-sensitive fluorescence probe Nile Red. A specific single-cysteine mutant of GroEL (Cys261), whose cysteine is located inside the central cavity at the apical region of the protein, was covalently labeled with synthetically prepared Nile Red maleimide (NR). Bulk fluorescence spectra of Cys261-NR were measured to examine the effects of binding of the stringent substrate, malate dehydrogenase (MDH), GroES, and nucleotide on the local environment of the probe. After binding denatured substrate, the fluorescence intensity increased by 32 +/- 7%, suggesting enhanced hydrophobicity at the position of the label. On the other hand, in the presence of ATP, the fluorescence intensity decreased by 13 +/- 3%, implying increased local polarity. To explore the sequence of local polarity changes, substrate, GroES, and various nucleotides were added in different orders; the resulting changes in emission intensity provide insight into the sequence of conformational changes occurring during GroEL-mediated protein folding. PMID- 16375457 TI - Complexation of lappaconitine with glycyrrhizic acid: stability and reactivity studies. AB - NMR and UV-vis spectroscopy have been used to study the complexation of antiarrhythmic alkaloid lappaconitine with an efficient complexing agent from licorice, glycyrrhizic acid, which is known to profoundly influence the therapeutic activity of the alkaloid in the complex. In MeOH, DMSO, or aqueous solutions, lappaconitine has been shown to form a stable complex with glycyrrhizic acid with 1:1 stoichiometry over a broad concentration range from 1 microM to 300 microM. The stability constant K(11) equals 2.0 x 10(5) M(-1) in aqueous solution. A similar complex of lappaconitine hydrobromide--the pharmaceutical formulation used in the treatment of arrhythmia--is 2 orders of magnitude less stable than pure lappaconitine. A notable decrease in the rate of the photoinduced electron-transfer reaction between lappaconitine in a complex with glycyrrhizic acid and tyrosine allows the suggestion of an explicit interrelation between the suppressed chemical reactivity of the bound alkaloid and the changes of its therapeutic efficiency. PMID- 16375458 TI - In situ time-resolved characterization of Au-CeO2 and AuOx-CeO2 catalysts during the water-gas shift reaction: presence of Au and O vacancies in the active phase. AB - Synchrotron-based in situ time-resolved x-ray diffraction and x-ray absorption spectroscopies were used to study the behavior of nanostructured {Au+AuO(x)} CeO(2) catalysts under the water-gas shift (WGS) reaction. At temperatures above 250 degrees C, a complete AuO(x)-->Au transformation was observed with high catalytic activity. Photoemission results for the oxidation and reduction of Au nanoparticles supported on rough ceria films or a CeO(2)(111) single crystal corroborate that cationic Au(delta+) species cannot be the key sites responsible for the WGS activity at high temperatures. The rate determining steps for the WGS seem to occur at the gold-ceria interface, with the active sites involving small gold clusters (<2 nm) and O vacancies. PMID- 16375459 TI - Transition from a Bloch-Wilson to a free-electron density of states in Zn(n)- clusters. AB - We present photoelectron spectroscopy studies on Zn(n) (-) in the size range of n=3-117. We show that zinc clusters exhibit a distinct transition in their electronic structure as a function of size. At small sizes (up to n=18) the clusters follow the Bloch-Wilson picture of the development of a metal from closed-shell atoms, exhibiting a gradual decrease of the gap between the fully occupied s band and the empty p band. For large sizes (n approximately or > 32) the band overlap allows the valence electrons to fully delocalize. This leads to an almost perfect free-electron density of states, as is demonstrated by discussing the spectra in the light of standard free-electron models and by comparison to the results obtained on sodium clusters. PMID- 16375460 TI - Grating-induced plasmon mode in gold nanoparticle arrays. AB - We study the dipolar coupling of gold nanoparticles arranged in regular two dimensional arrays by extinction micro-spectroscopy. When the interparticle spacing approaches the plasmon resonance wavelength of the individual particles, an additional band of very narrow width emerges in the extinction spectrum. By systematically changing the particles dielectric environment, the particles shape, the grating constant and angle of incidence, we show how this band associated to a grating induced-resonance can be influenced in strength and spectral position. The spectral position can be qualitatively understood by considering the conditions for grazing grating orders whereas the strength can be related to the strength of dipolar scattering from the individual particles. PMID- 16375461 TI - Excited state molecular structures and reactions directly determined by ultrafast electron diffraction. AB - In this communication, we report on the use of ultrafast electron diffraction to determine structural dynamics of excited states and reaction products of isolated aromatic carbonyls, acetophenone and benzaldehyde. For a 266 nm excitation, a bifurcation of pathways is structurally resolved, one leading to the formation of the triplet state (quinoid structure) and another to chemical products: for benzaldehyde the products are benzene and carbon monoxide (hydrogen migration and bond rupture) while those for acetophenone are the benzoyl and methyl radicals (bond rupture). The refined structures are compared with those predicted by theory. These dark structures and their radiationless transitions define the reduced energy landscape for complex reactions. PMID- 16375462 TI - Resonant coupling of bound excitons with LO phonons in ZnO: excitonic polaron states and Fano interference. AB - We report on a photoluminescence observation of robust excitonic polarons due to resonant coupling of exciton and longitudinal optical (LO) phonon as well as Fano type interference in high quality ZnO crystal. At low enough temperatures, resonant coupling of excitons and LO phonons leads to not only traditional Stokes lines (SLs) but also up to second-order anti-Stokes lines (ASLs) besides the zero phonon line (ZPL). The SLs and ASLs are found to be not mirror symmetric with respect to the ZPL, strongly suggesting that they are from different coupling states of exciton and phonons. Besides these spectral features showing the quasiparticle properties of exciton-phonon coupling system, the first-order SL is found to exhibit characteristic Fano lineshape, caused by quantum interference between the LO components of excitonic polarons and the continuous phonon bath. These findings lead to a new insight into fundamental effects of exciton-phonon interactions. PMID- 16375463 TI - Nuclear hyperfine interaction of rotating hydrogen: a spectroscopic investigation of hydrogen-OCS van der Waals complexes. AB - The rotational spectra of five weakly bonded hydrogen-OCS complexes (paraH(2), orthoH(2), HD, orthoD(2), and paraD(2)) are measured. Hyperfine structure is resolved and analyzed in all except the complex with paraH(2), where I=0. For the two j=1 species, orthoH(2)-OCS and paraD(2)-OCS, nuclear hyperfine coupling constants are found to be d(a)=21.2(2) and 8.4(2) kHz, respectively, indicative of nearly free uniaxial rotation of the hydrogen around the b-inertial axis. Similar analyses for HD-OCS and orthoD(2)-OCS yield the quadrupole coupling constants eqQ(a)=16(2) and 30(2) kHz, respectively, showing that the internal rotational motions of HD and orthoD(2) in the complex are slightly hindered producing a small nonspherical distribution. For orthoD(2)-OCS, the observed hyperfine structure indicates that the nuclear spin states I=0 and 2 are strongly coupled in the rotation of the complex. PMID- 16375464 TI - Efficient methods for finding transition states in chemical reactions: comparison of improved dimer method and partitioned rational function optimization method. AB - A combination of interpolation methods and local saddle-point search algorithms is probably the most efficient way of finding transition states in chemical reactions. Interpolation methods such as the growing-string method and the nudged elastic band are able to find an approximation to the minimum-energy pathway and thereby provide a good initial guess for a transition state and imaginary mode connecting both reactant and product states. Since interpolation methods employ usually just a small number of configurations and converge slowly close to the minimum-energy pathway, local methods such as partitioned rational function optimization methods using either exact or approximate Hessians or minimum-mode following methods such as the dimer or the Lanczos method have to be used to converge to the transition state. A modification to the original dimer method proposed by [Henkelman and Jonnson J. Chem. Phys. 111, 7010 (1999)] is presented, reducing the number of gradient calculations per cycle from six to four gradients or three gradients and one energy, and significantly improves the overall performance of the algorithm on quantum-chemical potential-energy surfaces, where forces are subject to numerical noise. A comparison is made between the dimer methods and the well-established partitioned rational function optimization methods for finding transition states after the use of interpolation methods. Results for 24 different small- to medium-sized chemical reactions covering a wide range of structural types demonstrate that the improved dimer method is an efficient alternative saddle-point search algorithm on medium-sized to large systems and is often even able to find transition states when partitioned rational function optimization methods fail to converge. PMID- 16375465 TI - Second-order Kohn-Sham perturbation theory: correlation potential for atoms in a cavity. AB - Second-order perturbation theory based on the Kohn-Sham Hamiltonian leads to an implicit density functional for the correlation energy E(c) (MP2), which is explicitly dependent on both occupied and unoccupied Kohn-Sham single-particle orbitals and energies. The corresponding correlation potential v(c) (MP2), which has to be evaluated by the optimized potential method, was found to be divergent in the asymptotic region of atoms, if positive-energy continuum states are included in the calculation [Facco Bonetti et al., Phys. Rev. Lett. 86, 2241 (2001)]. On the other hand, Niquet et al., [J. Chem. Phys. 118, 9504 (2003)] showed that v(c) (MP2) has the same asymptotic -alpha(2r(4)) behavior as the exact correlation potential, if the system under study has a discrete spectrum only. In this work we study v(c) (MP2) for atoms in a spherical cavity within a basis-set-free finite differences approach, ensuring a completely discrete spectrum by requiring hard-wall boundary conditions at the cavity radius. Choosing this radius sufficiently large, one can devise a numerical continuation procedure which allows to normalize v(c) (MP2) consistent with the standard choice v(c)(r-->infinity)=0 for free atoms, without modifying the potential in the chemically relevant region. An important prerequisite for the success of this scheme is the inclusion of very high-energy virtual states. Using this technique, we have calculated v(c) (MP2) for all closed-shell and spherical open-shell atoms up to argon. One finds that v(c) (MP2) reproduces the shell structure of the exact correlation potential very well but consistently overestimates the corresponding shell oscillations. In the case of spin-polarized atoms one observes a strong interrelation between the correlation potentials of the two spin channels, which is completely absent for standard density functionals. However, our results also demonstrate that E(c) (MP2) can only serve as a first step towards the construction of a suitable implicit correlation functional: The fundamental variational instability of this functional is recovered for beryllium, for which a breakdown of the self-consistent Kohn-Sham iteration is observed. Moreover, even for those atoms for which the self-consistent iteration is stable, the results indicate that the inclusion of v(c) (MP2) in the total Kohn-Sham potential does not lead to an improvement compared to the complete neglect of the correlation potential. PMID- 16375466 TI - Nonadiabatic chemical dynamics in an intense laser field: electronic wave packet coupled with classical nuclear motions. AB - Dynamics of molecules in an intense laser field is studied in terms of the quantum electronic wave packet coupled with classical nuclear motions. The equations of motion are derived taking a proper account of molecular interactions with the vector potential of a classical electromagnetic field, along with the nonadiabatic interaction due to the breakdown of the Born-Oppenheimer approximation. With the aid of electronic structure calculations, the present method enables us to track, in an ab initio manner, the dynamics of polyatomic molecules in an intense field. Preliminary calculations are carried out for the vibrational state of LiF and a collision of Li+F under an intense laser pulse, which are limited to the domain of no ionization. PMID- 16375467 TI - Quantum escape kinetics over a fluctuating barrier. AB - The escape rate of a particle over a fluctuating barrier in a double-well potential exhibits resonance at an optimum value of correlation time of fluctuation. This has been shown to be important in several variants of kinetic model of chemical reactions. We extend the analysis of this phenomenon of resonant activation to quantum domain to show how quantization significantly enhances resonant activation at low temperature due to tunneling. PMID- 16375468 TI - Renormalized coupled-cluster methods exploiting left eigenstates of the similarity-transformed Hamiltonian. AB - Completely renormalized (CR) coupled-cluster (CC) approaches, such as CR-CCSD(T), in which one corrects the standard CC singles and doubles (CCSD) energy for the effects of triply (T) and other higher-than-doubly excited clusters [K. Kowalski and P. Piecuch, J. Chem. Phys. 113, 18 (2000)], are reformulated in terms of the left eigenstates Phimid R:L of the similarity-transformed Hamiltonian of CC theory. The resulting CR-CCSD(T)(L) or CR-CC(2,3) and other CR-CC(L) methods are derived from the new biorthogonal form of the method of moments of CC equations (MMCC) in which, in analogy to the original MMCC theory, one focuses on the noniterative corrections to standard CC energies that recover the exact, full configuration-interaction energies. One of the advantages of the biorthogonal MMCC theory, which will be further analyzed and extended to excited states in a separate paper, is a rigorous size extensivity of the basic ground-state CR-CC(L) approximations that result from it, which was slightly violated by the original CR-CCSD(T) and CR-CCSD(TQ) approaches. This includes the CR-CCSD(T)(L) or CR CC(2,3) method discussed in this paper, in which one corrects the CCSD energy by the relatively inexpensive noniterative correction due to triples. Test calculations for bond breaking in HF, F(2), and H(2)O indicate that the noniterative CR-CCSD(T)(L) or CR-CC(2,3) approximation is very competitive with the standard CCSD(T) theory for nondegenerate closed-shell states, while being practically as accurate as the full CC approach with singles, doubles, and triples in the bond-breaking region. Calculations of the activation enthalpy for the thermal isomerizations of cyclopropane involving the trimethylene biradical as a transition state show that the noniterative CR-CCSD(T)(L) approximation is capable of providing activation enthalpies which perfectly agree with experiment. PMID- 16375469 TI - Adaptive resolution molecular-dynamics simulation: changing the degrees of freedom on the fly. AB - We present a new adaptive resolution technique for efficient particle-based multiscale molecular-dynamics simulations. The presented approach is tailor-made for molecular systems where atomistic resolution is required only in spatially localized domains whereas a lower mesoscopic level of detail is sufficient for the rest of the system. Our method allows an on-the-fly interchange between a given molecule's atomic and coarse-grained levels of description, enabling us to reach large length and time scales while spatially retaining atomistic details of the system. The new approach is tested on a model system of a liquid of tetrahedral molecules. The simulation box is divided into two regions: one containing only atomistically resolved tetrahedral molecules, and the other containing only one-particle coarse-grained spherical molecules. The molecules can freely move between the two regions while changing their level of resolution accordingly. The hybrid and the atomistically resolved systems have the same statistical properties at the same physical conditions. PMID- 16375470 TI - Influence of rotation and isotope effects on the dynamics of the N((2)D)+H(2) reactive system and of its deuterated variants. AB - Integral cross sections and thermal rate constants have been calculated for the N((2)D)+H(2) reaction and its isotopic variants N((2)D)+D(2) and the two-channel N((2)D)+HD by means of quasiclassical trajectory and statistical quantum mechanical model methods on the latest ab initio potential-energy surface [T.-S. Ho et al., J. Chem. Phys. 119, 3063 (2003)]. The effect of rotational excitation of the diatom on the dynamics of these reactions has been investigated and interesting discrepancies between the classical and statistical model calculations have been found. Whereas a net effect of reagent rotation on reactivity is always observed in the classical calculations, only a very slight effect is observed in the case of the asymmetric N((2)D)+HD reaction for the statistical quantum-mechanical method. The thermal rate constants calculated on this Potential-Energy Surface using quasiclassical trajectory and statistical model methods are in good agreement with the experimental determinations, although the latter are somewhat larger. A reevaluation of the collinear barrier of the potential surface used in the present study seems timely. Further theoretical and experimental studies are needed for a full understanding of the dynamics of the title reaction. PMID- 16375471 TI - Size-dependent charge-separation reaction for hydrated sulfate dianion cluster, SO(2-)(H2O)n, with n=3-7. AB - The decrease in the reaction rate for the charge separation in SO(4) (2 )(H(2)O)(n) with increasing cluster size is examined by first-principles calculations of the energetics, activation barriers, and thermal stability for n=3-7. The key factor governing the charge separation is the difference in the strength of solvation interaction: while interaction with water is strong for the reactant SO(4) (2-) and the product OH(-), it is relatively weak for HSO(4) (-). It gives rise to a barrier for charge separation as SO(4) (2-) is transformed into HSO(4) (-) and OH(-), although the overall reaction energy is exothermic. The barrier is high when more than two H(2)O are left to solvate HSO(4) (-), as in the case of symmetric solvation structure and in the case of large clusters. The entropy is another important factor since the potential surface is floppy and the thermal motion facilitates the symmetric distribution of H(2)O around SO(4) (2-), which leads to the gradual reduction in reaction rate and the eventual switch-off of charge separation as cluster size increases. The experimentally observed products for n=3-5 are explained by the thermally most favorable isomer at each size, obtained by ab initio molecular-dynamics simulations rather than by the isomer with the lowest energy. PMID- 16375472 TI - Observation of L uncoupling in the 5 1Deltag Rydberg state of Na2. AB - The phenomenon of electronic orbital angular momentum L uncoupled from its internuclear axis has been observed in the sodium dimer using high-resolution cw optical-optical double-resonance spectroscopy. When L uncoupling occurs, the degeneracy of Lambda doubling is removed. In our experiment, the intermediate B (1)Pi(u) state of Na(2) is excited from the thermally populated ground X (1)Sigma(g) (+) state by a single-line Ar(+) laser. Then, a single-mode dye laser is used to probe the Rydberg states from the intermediate state. The signals are detected by monitoring the UV fluorescence from the triplet gerade states back to the a (3)Sigma(u) (+) state via collision energy transfer. Under our experimental resolution, the splitting of Lambda doubling in the 5 (1)Delta(g) state of Na(2) can be measured. A total of 136 rovibronic levels with ef parities have been assigned to the 5 (1)Delta(g) state. The Lambda-splitting constants deduced from these data are q(0)=0.376(90)x10(-4) cm(-1), q(v)=0.114(6)x10(-4) cm(-1), and mu=0.76(33)x10(-8) cm(-1). In general, the Lambda splitting of the Delta states is considerably smaller than that of the Pi states. However, the first-order splitting constants q(0) and q(v) reported here are larger than those in the B (1)Pi(u) state. This is due to the L uncoupling of the Rydberg states. PMID- 16375473 TI - Molecular elimination in photolysis of o- and p-fluorotoluene at 193 nm: Internal energy of HF determined with time-resolved Fourier transform spectroscopy. AB - Following the photodissociation of o-fluorotoluene [o-C(6)H(4)(CH(3))F] at 193 nm, rotationally resolved emission spectra of HF(1< or =v< or =4) in the spectral region of 2800-4000 cm(-1) are detected with a step-scan Fourier transform spectrometer. HF(v< or =4) shows nearly Boltzmann-type rotational distributions corresponding to a temperature approximately 1080 K; a short extrapolation from data in the period of 0.5-4.5 mus leads to a nascent rotational temperature of 1130+/-100 K with an average rotational energy of 9+/-2 kJ mol(-1). The observed vibrational distribution of (v=1):(v=2):(v=3)=67.6: 23.2: 9.2 corresponds to a vibrational temperature of 5330+/-270 K. An average vibrational energy of 25+/ (3) (12) kJ mol(-1) is derived based on the observed population of HF(1< or =v< or =3) and estimates of the population of HF (v=0 and 4) by extrapolation. Experiments performed on p-fluorotoluene [p-C(6)H(4)(CH(3))F] yielded similar results with an average rotational energy of 9+/-2 kJ mol(-1) and vibrational energy of 26+/-(3) (12) kJ mol(-1) for HF. The observed distributions of internal energy of HF in both cases are consistent with that expected for four-center elimination. A modified impulse model taking into account geometries and displacement vectors of transition states during bond breaking predicts satisfactorily the rotational excitation of HF. An observed vibrational energy of HF produced from fluorotoluene slightly smaller than that from fluorobenzene might indicate the involvement of seven-membered-ring isomers upon photolysis. PMID- 16375474 TI - Differential cross sections for collisions of hexapole state-selected NO with He. AB - The first measurements of differential inelastic collision cross sections of fully state-selected NO (j=12, Omega=12, epsilon= -1) with He are presented. Full state selection is achieved by a 2 m long hexapole, which allows for a systematic study of the effect of parity conservation and breaking on the differential cross section. The collisionally excited NO molecules are detected using a resonant (1+1') REMPI ionization scheme in combination with the velocity-mapped, ion imaging technique. The current experimental configuration minimizes the contribution of noncolliding NO molecules in other rotational states j, Omega, epsilon--that contaminates images--and allows for study of the collision process at an unprecedented level of detail. A simple method to correct ion images for collision-induced alignment is presented as well and its performance is demonstrated. The present results show a significant difference between differential cross sections for scattering into the upper and lower component of the Lambda-doublet of NO. This result cannot be due to the energy splitting between these components. PMID- 16375475 TI - Effects of nuclear dynamics in the low-kinetic-energy Auger spectra of CO and CO2. AB - The CO and CO(2) carbon and oxygen Auger spectra have been measured by electron impact and compared with accurate theoretical calculations accounting for the effects of the dynamics of the nuclei on the energy and linewidth of the Auger bands. The calculations for CO were previously published [L. S. Cederbaum et al., J. Chem. Phys. 95, 6634 (1991)], while for CO(2) they are new and presented here for the first time. For both molecules, particular attention has been paid to the low-kinetic-energy region of the spectra, which corresponds to doubly charged ion states with the two holes mainly localized in the inner valence region. New bands have been observed. It is shown that a proper consideration of the vibrational broadening and shift of the bands due to the dynamics of the nuclei is needed to assign these features. For CO, very large energy shifts between corresponding features in the C 1s and O 1s spectra have been observed, confirming the theoretical predictions of 1991. The new computed spectra of CO(2) allow a very accurate analysis of the experiments over the whole energy range. PMID- 16375476 TI - Direct evidence for nonadiabatic dynamics in atom+polyatom reactions: crossed-jet laser studies of F+D2O-->DF+OD. AB - Quantum-state-resolved reactive-scattering dynamics of F+D(2)O-->DF+OD have been studied at E(c.m.)=5(1) kcal/mol in low-density crossed supersonic jets, exploiting pulsed discharge sources of F atom and laser-induced fluorescence to detect the nascent OD product under single-collision conditions. The product OD is formed exclusively in the v(OD)=0 state with only modest rotational excitation ( =0.50(1) kcal/mol), consistent with the relatively weak coupling of the 18.1(1) kcal/mol reaction exothermicity into "spectator" bond degrees of freedom. The majority of OD products [68(1)%] are found in the ground ((2)Pi(32) (+/-)) spin-orbit state, which adiabatically correlates with reaction over the lowest and only energetically accessible barrier (DeltaE( not equal) approximately 4 kcal/mol). However, 32(1)% of molecules are produced in the excited spin-orbit state ((2)Pi(12) (+/-)), although from a purely adiabatic perspective, this requires passage over a DeltaE( not equal) approximately 25 kcal/mol barrier energetically inaccessible at these collision energies. This provides unambiguous evidence for nonadiabatic surface hopping in F+D(2)O atom abstraction reactions, indicating that reactive-scattering dynamics even in simple atom+polyatom systems is not always isolated on the ground electronic surface. Additionally, the nascent OD rotational states are well fitted by a two-temperature Boltzmann distribution, suggesting correlated branching of the reaction products into the DF(v=2,3) vibrational manifold. PMID- 16375477 TI - Photodetachment spectroscopy of PtBr4(2-): probing the Coulomb barrier of a doubly charged anion. AB - We probe the repulsive Coulomb barrier of the doubly charged anion PtBr(4) (2-) by photodetachment spectroscopy. The results are discussed in terms of models for the photoemission process, the excitation spectrum of PtBr(4) (2-), and calculations of the energy-dependent tunneling probability for various model potentials. PMID- 16375478 TI - First observation of a dihydrogen bond involving the Si-H group in phenol diethylmethylsilane clusters by infrared-ultraviolet double-resonance spectroscopy. AB - We have experimentally identified a dihydrogen bond involving the Si-H group in phenol-diethylmethylsilane (DEMS) clusters for the first time by IR-UV double resonance spectroscopy. Vibrational shifts to lower frequency of 21-29 cm(-1) were found for the OH stretching vibration of three isomers of the phenol-DEMS clusters. Spectral simulations based on the MP2 calculations also support our observation. In addition to these clusters, dihydrogen bonds were also observed in the phenol-H(2)O-DEMS and (phenol)(2)-DEMS clusters, which exhibited much stronger interactions than the phenol-DEMS clusters. PMID- 16375479 TI - Multidimensional infrared spectroscopy for molecular vibrational modes with dipolar interactions, anharmonicity, and nonlinearity of dipole moments and polarizability. AB - We present an analytical expression for the linear and nonlinear infrared spectra of interacting molecular vibrational motions. Each of the molecular modes is explicitly represented by a classical damped oscillator on an anharmonic multidimensional potential-energy surface. The two essential interactions, the dipole-dipole (DD) and the dipole-induced-dipole (DID) interactions, are taken into account, and each dipole moment and polarizability are expanded to nonlinear order with respect to the nuclear vibrational coordinate. Our analytical treatment leads to expressions for the contributions of anharmonicity, DD and DID interactions, and the nonlinearity of dipole moments and polarizability elements to the one-, two-, and three-dimensional spectra as separated terms, which allows us to discuss the relative importance of these respective contributions. We can calculate multidimensional signals for various configurations of molecules interacting through DD and DID interactions for different material parameters over the whole range of frequencies. We demonstrate that contributions from the DD and DID interactions and anharmonicity are separately detectable through the third-order three-dimensional IR spectroscopy, whereas they cannot be distinguished from each other in either the linear or the second-order IR spectroscopies. The possibility of obtaining the intra- or intermolecular structural information from multidimensional spectra is also discussed. PMID- 16375480 TI - Determination of the excited-state structure of 7-azaindole-water cluster using a Franck-Condon analysis. AB - The change of the 7-azaindole-water cluster structure upon electronic excitation was determined by a Franck-Condon analysis of the intensities in the fluorescence emission spectra obtained via excitation of five different vibronic bands. A total of 105 emission band intensities were fitted, together with the changes of rotational constants of one isotopomer. These rotational constants have been obtained from a fit to the rovibronic contour of the cluster. The geometry change upon electronic excitation to the pi pi* state can be described by a strong and asymmetric shortening of the hydrogen bonds and a deformation of both the pyridine and the pyrrole rings of 7-azaindole. The resulting geometry changes are interpreted on the basis of ab initio calculations. PMID- 16375481 TI - Small gas-phase dianions produced by sputtering and gas flooding. AB - We have extended our previous experiment [Schauer et al., Phys. Rev. Lett. 65, 625 (1990)] where we had produced small gas-phase dianion clusters of C(n) (2-)(n > or =7) by means of sputtering a graphite surface by Cs(+) ion bombardment. Our detection sensitivity for small C(n) (2-) could now be increased by a factor of about 50 for odd n. Nevertheless, a search for the elusive pentamer dianion of C(5) (2-) was not successful. As an upper limit, the sputtered flux of C(5) (2-) must be at least a factor of 5000 lower than that of C(7) (2-), provided that the lifetime of C(5) (2-) is sufficiently long to allow its detection by mass spectrometry. When oxygen gas (flooding with either O(2) or with N(2)O) was supplied to the Cs(+)-bombarded graphite surface, small dianions of OC(n) (2-)(5< or =n < or =14) and O(2)C(7) (2-) were observed in addition to C(n) (2-)(n > or =7). Similarly, Cs(+) sputtering of graphite with simultaneous SF(6) gas flooding produced SC(n) (2-)(6< or =n< or =18). Mixed nitrogen-carbon or fluorine-carbon dianion clusters could not be observed by these means. Attempts to detect mixed metal-fluoride dianions for SF(6) gas flooding of various Cs(+)-bombarded metal surfaces were successful for the case of Zr, where metastable ZrF(6) (2-) was observed. Cs(+) bombardment of a silicon carbide (SiC) wafer produced SiC(n) (2-) (n=6,8,10). When oxygen gas was supplied to the Cs(+)-bombarded SiC surface, small dianions of SiOC(n) (2-) (n=4,6,8) and of SiO(2)C(n) (2-) (n=4,6) as well as a heavier unidentified dianion (at mz=98.5) were observed. For toluene (C(7)H(8)) vapor flooding of a Cs(+)-bombarded graphite surface, several hydrocarbon dianion clusters of C(n)H(m) (2-)(n> or =7) were produced in addition to C(n) (2-)(n> or =7), while smaller C(n)H(m) (2-) with n< or =6 could not be observed. BeC(n) (2-) (n=4,6,8,10), Be(2)C(6) (2-), as well as BeC(8)H(m) (2-) (with m=2 and/or m=1) were observed for toluene vapor flooding of a Cs(+) bombarded beryllium metal foil. The metastable pentamer (9)Be(12)C(4) (2-) at mz=28.5 was the smallest and lightest dianion molecule that we could detect. The small dianion clusters of SC(n) (2-), OC(n) (2-), BeC(n) (2-), and SiO(m)C(n) (2 ) (m=0,1,2) have different abundance patterns. A resemblance exists between the abundance patterns of BeC(n) (2-) and SiC(n) (2-), even though calculated molecular structures of BeC(6) (2-) and SiC(6) (2-) are different. The abundance pattern of SC(n) (2-) is fairly similar to that of C(n) (2-). PMID- 16375482 TI - (HCl)2 and (HF)2 in small helium clusters: quantum solvation of hydrogen-bonded dimers. AB - We present a rigorous theoretical study of the solvation of (HCl)(2) and (HF)(2) by small ((4)He)(n) clusters, with n=1-14 and 30. Pairwise-additive potential energy surfaces of He(n)(HX)(2) (X=Cl and F) clusters are constructed from highly accurate four-dimensional (rigid monomer) HX-HX and two-dimensional (rigid monomer) He-HX potentials and a one-dimensional He-He potential. The minimum energy geometries of these clusters, for n=1-6 in the case of (HCl)(2) and n=1-5 for (HF)(2), correspond to the He atoms in a ring perpendicular to and bisecting the HX-HX axis. The quantum-mechanical ground-state energies and vibrationally averaged structures of He(n)(HCl)(2) (n=1-14 and 30) and He(n)(HF)(2) (n=1-10) clusters are calculated exactly using the diffusion Monte Carlo (DMC) method. In addition, the interchange-tunneling splittings of He(n)(HCl)(2) clusters with n=1 14 are determined using the fixed-node DMC approach, which was employed by us previously to calculate the tunneling splittings for He(n)(HF)(2) clusters, n=1 10 [A. Sarsa et al., Phys. Rev. Lett. 88, 123401 (2002)]. The vibrationally averaged structures of He(n)(HX)(2) clusters with n=1-6 for (HCl)(2) and n=1-5 for (HF)(2) have the helium density localized in an effectively one-dimensional ring, or doughnut, perpendicular to and at the midpoint of the HX-HX axis. The rigidity of the solvent ring varies with n and reaches its maximum for the cluster size at which the ring is filled, n=6 and n=5 for (HCl)(2) and (HF)(2), respectively. Once the equatorial ring is full, the helium density spreads along the HX-HX axis, eventually solvating the entire HX dimer. The interchange tunneling splitting of He(n)(HCl)(2) clusters hardly varies at all over the cluster size range considered, n=1-14, and is virtually identical to that of the free HCl dimer. This absence of the solvent effect is in sharp contrast with our earlier results for He(n)(HF)(2) clusters, which show a approximately 30% reduction of the tunneling splitting for n=4. A tentative explanation for this difference is proposed. The implications of our results for the interchange tunneling dynamics of (HCl)(2) in helium nanodroplets are discussed. PMID- 16375483 TI - Water-carbon dioxide mixtures at high temperatures and pressures: local order in the water rich phase investigated by vibrational spectroscopy. AB - Raman scattering combined with near- and mid-infrared absorption spectroscopies was used to investigate the evolution of the local order in the water rich phase of water-CO(2) mixtures under isobaric heating (T=40-360 degrees C, P=250 bars). The quantitative analysis of the spectra shows that tetramers and larger oligomers are the main constituents of water at moderate temperatures below 80 degrees C. As the temperature increases, the dimer and trimer concentrations considerably increase at the expense of larger oligomers. Finally, water dimers are predominant at the highest temperature investigated close to the temperature of total miscibility of the mixture (T=366 degrees C, P=250 bars). This result is consistent with our previous investigation [R. Oparin T. Tassaing, Y. Danten, and M. Besnard, J. Chem. Phys. 120, 10691 (2004)] on water dissolved in the CO(2) rich phase where we found that close to the temperature of total miscibility water also exists mainly under dimeric form. The current study combined with that mentioned above provides a model investigation of the evolution of the state of aggregation of water molecules in binary mixture involving a hydrophobic solvent in a wide range of temperature. PMID- 16375484 TI - The reorganization energy of electron transfer in nonpolar solvents: molecular level treatment of the solvent. AB - The intermolecular electron transfer in a solute pair consisting of pyrene and dimethylaniline is investigated in a nonpolar solvent, n-hexane. The earlier elaborated approach [M. Tachiya, J. Phys Chem. 97, 5911 (1993)] is used; this method provides a physically relevant background for separating inertial and inertialess polarization responses for both nonpolarizable and polarizable molecular level simulations. The molecular-dynamics technique was implemented for obtaining the equilibrium ensemble of solvent configurations. The nonpolar solvent, n-hexane, was treated in terms of OPLS-AA parametrization. Solute Lennard-Jones parameters were taken from the same parametrization. Solute charge distributions of the initial and final states were determined using ab initio level [HF/6-31G(d,p)] quantum-chemical calculations. Configuration analysis was performed explicitly taking into account the anisotropic polarizability of n hexane. It is shown that the Gaussian law well describes calculated distribution functions of the solvent coordinate, therefore, the rate constant of the ET reaction can be characterized by the reorganization energy. Evaluated values of the reorganization energies are in a range of 0.03-0.11 eV and significant contribution (more then 40% of magnitude) comes from anisotropic polarizability. Investigation of the reorganization energy lambda dependence on the solute pair separation distance d revealed unexpected behavior. The dependence has a very sharp peak at the distance d=7 A where solvent molecules are able to penetrate into the intermediate space between the solute pair. The reason for such behavior is clarified. This new effect has a purely molecular origin and cannot be described within conventional continuum solvent models. PMID- 16375485 TI - NMR probing of structural peculiarities in ionic solutions close to critical point. AB - (1)H, (23)Na, (35)Cl, (79)Br, and (81)Br NMR chemical shifts (delta) and signal half widths (Delta(12)) have been measured in aqueous electrolyte mixtures [tetrahydrofuran/H(2)ONaCl and 3-methylpyridine (3MP)H(2)ONaBr] at different mass fractions of salt (X) in the one-phase region, close to their lower critical solution points (T(CL)). Discontinuous changes in slope of delta=f(X) and Delta(12)=f(X) have been found in (23)Na and (81)Br NMR spectra of 3MP/water/NaBr solution at X approximately 0.1 and T=301 K. The dependency of (1)H NMR signals of 3MP is continuous over the whole investigated range of X=0.002-0.2, whereas changes in the slope of H(2)O chemical shifts are hardly noticeable. In the two phase region, i.e., at T>T(CL), a doubling of all NMR signals has been observed. The sensitivity of NMR parameters depends more on composition of solution for anions (Cl(-) and Br(-)) than for cations (Na(+)). A very strong relaxation effect for (81)Br nuclei with relaxation rates reaching 14 000 s(-1) was observed. The results are interpreted in terms of ion-molecular clustering and changes in coherency of dipole configurations of water molecules during supramolecular restructuring of solutions. PMID- 16375486 TI - Properties of microfluidic turbulent mixing revealed by fluorescence lifetime imaging. AB - We present a new method of measurement based on fluorescence lifetime imaging that reveals molecular-scale details of the mixing process in a continuous-flow turbulent microfluidic reactor. Our data provide a glimpse of the cascade to the minimal eddy size, followed by rapid diffusion involving the smallest eddies for final mixing. PMID- 16375487 TI - Dynamic salt effect on intramolecular charge-transfer reactions. AB - The dynamic salt effect in charge-transfer reactions is investigated theoretically in this paper. Free-energy surfaces are derived based on a nonequilibrium free-energy functional. Reaction coordinates are clearly defined. The solution of the reaction-diffusion equation leads to a rate constant depending on the time correlation function of the reaction coordinates. The time correlation function of the ion-atmosphere coordinate is derived from the solution of the Debye-Falkenhagen equation. It is shown that the dynamic salt effect plays an important role in controlling the rate of charge-transfer reactions in the narrow-window limit but is balanced by the energetics and the dynamics of the polar-solvent coordinate. The simplest version of the theory is compared with an experiment, and the agreement is fairly good. The theory can also be extended to charge-transfer in the class of electrolytes that has come to be called "ionic fluids." PMID- 16375488 TI - Heterogeneous thermal excitation and relaxation in supercooled liquids. AB - We investigate a phenomenological model which rationalizes the effects of dielectric hole burning on the basis of heterogeneous dielectric and specific heat relaxation in supercooled liquids. The quantitative agreement between model predictions and dielectric hole-burning observations is lost if the assumption of correlated dielectric and thermal relaxation times is removed from the model. This suggests that dynamically distinct domains in real liquids are associated with a time constant which characterizes both the structural and thermal relaxation behaviors. The calculations demonstrate that the observed burn-induced modifications reflect the spectral selectivity and persistence time of the fictive temperatures within these domains, and that 100 or more cycles of the sinusoidal burn field can be required to saturate the heat accumulated in the slow degrees of freedom. It is also shown that the recovery of dielectric holes is entirely accounted for by the model, and that the persistence times do not provide direct insight into rate exchange processes. Additionally, the model predicts that the heating effects considered here are a significant source of nonlinear dielectric behavior, even in the absence of deliberate frequency selective hole burning. PMID- 16375489 TI - A study of two-electron quantum dot spectrum using discrete variable representation method. AB - A variational method called discrete variable representation is applied to study the energy spectra of two interacting electrons in a quantum dot with a three dimensional anisotropic harmonic confinement potential. This method, applied originally to problems in molecular physics and theoretical chemistry, is here used to solve the eigenvalue equation to relative motion between the electrons. The two-electron quantum dot spectrum is determined then with a precision of at least six digits. Moreover, the electron correlation energies for various potential confinement parameters are investigated for singlet and triplet states. When possible, the present results are compared with the available theoretical values. PMID- 16375490 TI - Structure and dynamics of water at the interface with phospholipid bilayers. AB - We have performed two molecular-dynamics simulations to study the structural and dynamical properties of water at the interface with phospholipid bilayers. In one of the simulations the bilayer contained neutral phospholipid molecules, dioleoylphosphatidylcholine (DOPC); in the second simulation the bilayer contained charged lipid molecules, dioleoylphosphatidylserine (DOPS). From the density profile of water we observe that water next to the DOPS bilayer is more perturbed as compared to water near the DOPC bilayer. Using an energetic criterion for the determination of hydrogen bonding we find that water molecules create strong hydrogen bonds with the headgroups of the phospholipid molecules. Due to the presence of these bonds and also due to the confinement of water, the translational and orientational dynamics of water at the interface are slowed down. The degree of slowing down of the dynamics depends upon the location of water molecules near a lipid headgroup. PMID- 16375491 TI - Kinetics of CO oxidation on high-concentration phases of atomic oxygen on Pt(111). AB - Temperature-programmed reaction spectroscopy (TPRS) and direct, isothermal reaction-rate measurements were employed to investigate the oxidation of CO on Pt(111) covered with high concentrations of atomic oxygen. The TPRS results show that oxygen atoms chemisorbed on Pt(111) at coverages just above 0.25 ML (monolayers) are reactive toward coadsorbed CO, producing CO(2) at about 295 K. The uptake of CO on Pt(111) is found to decrease with increasing oxygen coverage beyond 0.25 ML and becomes immeasurable at a surface temperature of 100 K when Pt(111) is partially covered with Pt oxide domains at oxygen coverages above 1.5 ML. The rate of CO oxidation measured as a function of CO beam exposure to the surface exhibits a nearly linear increase toward a maximum for initial oxygen coverages between 0.25 and 0.50 ML and constant surface temperatures between 300 and 500 K. At a fixed CO incident flux, the time required to reach the maximum reaction rate increases as the initial oxygen coverage is increased to 0.50 ML. A time lag prior to the reaction-rate maximum is also observed when Pt oxide domains are present on the surface, but the reaction rate increases more slowly with CO exposure and much longer time lags are observed, indicating that the oxide phase is less reactive toward CO than are chemisorbed oxygen atoms on Pt(111). On the partially oxidized surface, the CO exposure needed to reach the rate maximum increases significantly with increases in both the initial oxygen coverage and the surface temperature. A kinetic model is developed that reproduces the qualitative dependence of the CO oxidation rate on the atomic oxygen coverage and the surface temperature. The model assumes that CO chemisorption and reaction occur only on regions of the surface covered by chemisorbed oxygen atoms and describes the CO chemisorption probability as a decreasing function of the atomic oxygen coverage in the chemisorbed phase. The model also takes into account the migration of oxygen atoms from oxide domains to domains with chemisorbed oxygen atoms. According to the model, the reaction rate initially increases with the CO exposure because the rate of CO chemisorption is enhanced as the coverage of chemisorbed oxygen atoms decreases during reaction. Longer rate delays are predicted for the partially oxidized surface because oxygen migration from the oxide phase maintains high oxygen coverages in the coexisting chemisorbed oxygen phase that hinder CO chemisorption. It is shown that the time evolution of the CO oxidation rate is determined by the relative rates of CO chemisorption and oxygen migration, R(ad) and R(m), respectively, with an increase in the relative rate of oxygen migration acting to inhibit the reaction. We find that the time lag in the reaction rate increases nearly exponentially with the initial oxygen coverage [O](i) (tot) when [O](i) (tot) exceeds a critical value, which is defined as the coverage above which R(ad)R(m) is less than unity at fixed CO incident flux and surface temperature. These results demonstrate that the kinetics for CO oxidation on oxidized Pt(111) is governed by the sensitivity of CO binding and chemisorption on the atomic oxygen coverage and the distribution of surface oxygen phases. PMID- 16375492 TI - Effects of dielectric saturation and ionic screening on the proton self-diffusion coefficients in perfluorosulfonic acid membranes. AB - Proton transport in perfluorosulfonic acid (PFSA) membranes is investigated through a statistical mechanical model that includes the effects of the interaction of the tethered sulfonate groups with both the water and solvated protons. We first derive a potential that describes the electrostatic field due to the dissociated sulfonic acid groups by extending the work of Gronbech-Jensen et al. [ Mol. Phys. 92, 941 (1997)] to a finite array of point charges. A highly convergent series is obtained which includes the effects of screening due to the protons. We then investigate the effects of both dielectric saturation and two distinct formulations of ionic screening on the proton self-diffusion coefficient in Nafion membranes over a range of water contents. Our computations show that the two phenomena (i.e., dielectric saturation and ionic screening) under constant temperature conditions result in canceling affects. Our calculations provide a radial dependence of the proton mobility suggesting that the dominant self-diffusion occurs in the central region of the pores, well separated from the sulfonate groups. Through comparison of our calculated diffusion coefficients with the experimental values we derived a slightly smaller average separation distance of the hydronium ion from the sulfonate ions than suggested by either electronic structure calculations or multistate empirical valence bond molecular dynamics simulations. PMID- 16375493 TI - Effect of nanoconfinement on liquid-crystal polymer chains. AB - We apply a Monte Carlo polymerization model for Gay-Berne [J. Chem. Phys. 74, 3316 (1981)] monomers that we have recently introduced [J. Chem. Phys. 121, 9123 (2004)] to investigate with computer simulations the effects of nanoconfinement and anchoring type on the structure of the main-chain liquid-crystal polymers formed in thin films, in the presence of several types of surface alignment: parallel to the interface (random and uniform) or perpendicular to it (homeotropic). We perform first a study of the confined monomers and then we examine the features of the polymer chains obtained from an isotropic or nematic sample. We find a significant effect of the anchoring conditions on the characteristics of the chains and particularly striking differences between planar and homeotropic boundaries. Furthermore, our results indicate that the choice of different anchorings could be used to tune the linearity and degree of polymerization of the chains. PMID- 16375494 TI - Modeling the flow of fluid/particle mixtures in microchannels: encapsulating nanoparticles within monodisperse droplets. AB - We develop a hybrid computational approach for simulating mixtures of binary fluids and mobile, submicron particles. The model couples a lattice Boltzmann method for the binary fluid with a Brownian dynamics model for the particles. The particles can exhibit preferential wetting interactions with the different components of the fluid. As an illustration of the method, we carry out simulations in two dimensions to compare the spinodal decomposition of a pure binary fluid with the phase separation of binary blends that contain either fixed or mobile particles. We then isolate conditions where the flow of a binary fluid/particle mixture past surfaces with well-defined asperities leads to the formation of monodisperse droplets, which encapsulate the nanoparticles. The findings provide guidelines for creating multiphase emulsions with well controlled morphologies. PMID- 16375495 TI - Structure determination of CdS and ZnS nanoparticles: direct modeling of synchrotron-radiation diffraction data. AB - We introduce a modified method of powder-diffraction data analysis to obtain precise structural information on freestanding ZnS and CdS nanoparticles with diameters well below 5 nm, i.e., in a range where common bulk-derived approaches fail. The method is based on the Debye equation and allows us to access the crystal structure and the size of the particles with high precision. Detailed information on strain, relaxation effects, stacking faults, and the shape of the particles becomes available. We find significant size differences between our new results and those obtained by established methods, and conclude that a mixed zinc blende/wurtzite stacking and significant lattice distortions occur in our CdS nanoparticles. Our approach should have direct impact on the understanding and modeling of quantum size effects in nanoparticles. PMID- 16375496 TI - Long time scale blinking kinetics of cyanine fluorophores conjugated to DNA and its effect on Forster resonance energy transfer. AB - The blinking kinetics of individual Cy5 fluorophores conjugated to DNA are directly measured using single-molecule spectroscopy. Under deoxygenated aqueous conditions, Cy5 fluorescence exhibits spontaneous and reversible on/off fluctuations with a period lasting seconds. This blinking is observed when directly exciting Cy5 with 640 nm light and by Forster resonance energy transfer (FRET). We find that Cy5 blinking is influenced by the proximity of the donor, the structure of the donor, the presence of 514 nm excitation, and FRET. In the context of single-molecule FRET, blinking of the acceptor produces anticorrelated donor-acceptor intensity fluctuations, which can be difficult to discern from variations in the interdye distance. Slow blinking is, in particular, problematic because it overlaps with biologically relevant time scales. By employing an alternating 514640 nm laser excitation scheme, we show that the dark states can be readily resolved and discriminated from FRET distance fluctuations. PMID- 16375497 TI - New insights on the stereodynamics of ethylene adsorption on an oxygen-precovered silver surface. AB - The control of spatial orientation of molecules has a great influence on the stereodynamics of elementary processes occurring both in homogeneous and heterogeneous phases. Nonpolar molecules have so far escaped direct experimental investigations because of their poor sensitivity to several external constraints. Recently, it has been shown that the collisional alignment produced in supersonic expansions coupled with molecular-beam velocity selection can help solve such problems. Here we show that the sticking probability of ethylene, a nonpolar molecule prototypical of unsaturated hydrocarbons, on an O(2)-precovered Ag(001) surface is larger for molecules approaching in a helicopter-like motion than for those cartwheeling. A mechanism involving a weakly bound precursor state is suggested, with helicopter molecules having a lower chance of being scattered back into the gas phase than cartwheels when colliding with preadsorbed ethylene. PMID- 16375498 TI - Solvent-free model for self-assembling fluid bilayer membranes: stabilization of the fluid phase based on broad attractive tail potentials. AB - We present a simple and highly adaptable method for simulating coarse-grained lipid membranes without explicit solvent. Lipids are represented by one head bead and two tail beads, with the interaction between tails being of key importance in stabilizing the fluid phase. Two such tail-tail potentials were tested, with the important feature in both cases being a variable range of attraction. We examined phase diagrams of this range versus temperature for both functional forms of the tail-tail attraction and found that a certain threshold attractive width was required to stabilize the fluid phase. Within the fluid-phase region we find that material properties such as area per lipid, orientational order, diffusion constant, interleaflet flip-flop rate, and bilayer stiffness all depend strongly and monotonically on the attractive width. For three particular values of the potential width we investigate the transition between gel and fluid phases via heating or cooling and find that this transition is discontinuous with considerable hysteresis. We also investigated the stretching of a bilayer to eventually form a pore and found excellent agreement with recent analytic theory. PMID- 16375499 TI - Prediction of molecular-dynamics simulation results using feedforward neural networks: reaction of a C2 dimer with an activated diamond (100) surface. AB - A new approach involving neural networks combined with molecular dynamics has been used for the determination of reaction probabilities as a function of various input parameters for the reactions associated with the chemical-vapor deposition of carbon dimers on a diamond (100) surface. The data generated by the simulations have been used to train and test neural networks. The probabilities of chemisorption, scattering, and desorption as a function of input parameters, such as rotational energy, translational energy, and direction of the incident velocity vector of the carbon dimer, have been considered. The very good agreement obtained between the predictions of neural networks and those provided by molecular dynamics and the fact that, after training the network, the determination of the interpolated probabilities as a function of various input parameters involves only the evaluation of simple analytical expressions rather than computationally intensive algorithms show that neural networks are extremely powerful tools for interpolating the probabilities and rates of chemical reactions. We also find that a neural network fits the underlying trends in the data rather than the statistical variations present in the molecular-dynamics results. Consequently, neural networks can also provide a computationally convenient means of averaging the statistical variations inherent in molecular dynamics calculations. In the present case the application of this method is found to reduce the statistical uncertainty in the molecular-dynamics results by about a factor of 3.5. PMID- 16375500 TI - Charge transfer in FeO: a combined molecular-dynamics and ab initio study. AB - Molecular-dynamics simulations and ab initio electronic structure calculations were carried out to determine the rate of charge transfer in stoichiometric wustite (FeO). The charge transfer of interest occurs by II/III valence interchange between nearest-neighbor Fe atoms, with the Fe(III) constituting a "hole" electronic defect. There are two possible nearest-neighbor charge transfers in the FeO lattice, which occur between edge-sharing or corner-sharing FeO(6) octahedra. Molecular-dynamics simulations predict charge-transfer rates of 3.7 x 10(11) and 1.9 x 10(9) s(-1) for the edge and corner transfers, respectively, in good agreement with those calculated using an ab initio cluster approach (1.6 x 10(11) and 8.0 x 10(8) s(-1), respectively). The calculated rates are also similar to those along the basal and c-axis directions in hematite (alpha-Fe(2)O(3)) determined previously. Therefore, as is the case for hematite, wustite is predicted to show anisotropic electrical conductivity. Our findings indicate that a rigid-ion model does not give acceptable results, thus showing the need to account for the change in polarizability of the system upon charge transfer. Our model achieves this by using a simple mechanical shell model. By calculating the electronic coupling matrix elements for many transition state configurations obtained from the molecular-dynamics simulations, we found evidence that the position of the bridging oxygen atoms can greatly affect the amount of electronic coupling between the donor and acceptor states. Finally, we address the effect of oxygen vacancies on the charge transfer. It was found that an oxygen vacancy not only creates a driving force for holes to transport away from the vacancy (or equivalently for electrons to diffuse toward the vacancy) but also lowers the free-energy barriers for charge transfer. In addition, the reorganization energy significantly differed from the nondefective case in a small radius around the defect. PMID- 16375501 TI - Reconsideration of second-harmonic generation from isotropic liquid interface: broken Kleinman symmetry of neat air/water interface from dipolar contribution. AB - It has been generally accepted that there are significant quadrupolar and bulk contributions to the second-harmonic generation (SHG) reflected from the neat air/water interface, as well as common liquid interfaces. Because there has been no general methodology to determine the quadrupolar and bulk contributions to the SHG signal from a liquid interface, this conclusion was reached based on the following two experimental phenomena: the breaking of the macroscopic Kleinman symmetry and the significant temperature dependence of the SHG signal from the neat air/water interface. However, because the sum frequency generation vibrational spectroscopy (SFG-VS) measurement of the neat air/water interface observed no apparent temperature dependence, the temperature dependence in the SHG measurement has been reexamined and proven to be an experimental artifact. Here we present a complete microscopic analysis of the susceptibility tensors of the air/water interface, and show that dipolar contribution alone can be used to address the issue of the breaking of the macroscopic Kleinman symmetry at the neat air/water interface. Using this analysis, the orientation of the water molecules at the interface can be obtained, and it is consistent with the measurement from SFG-VS. Therefore, the key rationales to conclude significantly quadrupolar and bulk contributions to the SHG signal of the neat air/water interface can no longer be considered as valid as before. This new understanding of the air/water interface can shed light on our understanding of the nonlinear optical responses from other molecular interfaces as well. PMID- 16375502 TI - Surface-induced C-O bond anharmonicity of methoxy adsorbed on Cu(100): experiments and density-functional theory calculations. AB - The anharmonic properties of a surface intermediate, methoxy, adsorbed on Cu(100) are investigated by surface infrared overtone spectroscopy and density-functional theory electronic structure calculations. The anharmonicity is measured in the zero-coverage limit, and it is observed that the anharmonicity is increased upon adsorption as compared with the free methanol. By combining experiments with calculations we demonstrate that modifications of the anharmonicity of the methoxy species is indeed induced by adsorption onto the copper surface and not by the formation of the methoxy species. PMID- 16375503 TI - Photoluminescence and spectral holeburning in europium-doped MgS nanoparticles. AB - Luminescence and power-gated spectral holeburning studies have been performed on Eu-doped MgS nanoparticles. These particles are atomically tailored to produce and control the relative concentration of Eu(2+) and Eu(3+), which is necessary for power-gated holeburning. The spectral holes are permanent at low temperatures. Optical studies show that the electron-phonon coupling is stronger in nanoparticles than in thin films or microparticles of the same material. This is the reason for inherently broader spectral holes in nanoparticles as compared to microparticle or thin-film samples. Temperature broadening of spectral holes in nanoparticles follows a T(2.4) behavior, a faster rate than thin films or microparticles. This behavior can be attributed to the glassy nature of the particles produced. PMID- 16375504 TI - Theory of glassy dynamics in conformationally anisotropic polymer systems. AB - A mode coupling theory for the ideal glass transition temperature, or crossover temperature to highly activated dynamics in the deeply supercooled regime, T(c), has been developed for anisotropic polymer liquids. A generalization of a simplified mode coupling approach at the coarse-grained segment level is employed which utilizes structural and thermodynamic information from the anisotropic polymer reference interaction site model theory. Conformational alignment or/and coil deformation modifies equilibrium properties and constraining interchain forces thereby inducing anisotropic segmental dynamics. For liquid-crystalline polymers a small suppression of T(c) with increasing nematic or discotic orientational order is predicted. The underlying mechanism is reduction of the degree of coil interpenetration and intermolecular repulsive contacts due to segmental alignment. For rubber networks chain deformation results in an enhanced bulk modulus and a modest elevation of T(c) is predicted. The theory can also be qualitatively applied to systems that undergo nonuniversal local deformation and alignment, such as polymer thin films and grafted brush layers, and large elevations or depressions of T(c) are possible. Extension to treat directionally dependent collective barrier formation and activated hopping is possible. PMID- 16375505 TI - Domain growth, budding, and fission in phase-separating self-assembled fluid bilayers. AB - A systematic investigation of the phase-separation dynamics in self-assembled binary fluid vesicles and open membranes is presented. We use large-scale dissipative particle dynamics to explicitly account for solvent, thereby allowing for numerical investigation of the effects of hydrodynamics and area-to-volume constraints. In the case of asymmetric lipid composition, we observed regimes corresponding to coalescence of flat patches, budding, vesiculation, and coalescence of caps. The area-to-volume constraint and hydrodynamics have a strong influence on these regimes and the crossovers between them. In the case of symmetric mixtures, irrespective of the area-to-volume ratio, we observed a growth regime with an exponent of 1/2. The same exponent is also found in the case of open membranes with symmetric composition. PMID- 16375506 TI - Scaling of mesoscale simulations of polymer melts with the bare friction coefficient. AB - Both the Rouse and reptation model predict that the dynamics of a polymer melt scale inversely proportional with the Langevin friction coefficient xi. Mesoscale Brownian dynamics simulations of polyethylene validate these scaling predictions, providing the reptational friction xi(R)=xi+xi(C) is used, where xi(C) reflects the fundamental difference between a deterministic and a stochastic propagator even in the limit of xi to zero. The simulations have been performed with Langevin background friction and with pairwise friction, as in dissipative particle dynamics. Both simulation methods lead to equal scaling behavior with xi(C) having almost the same value in both cases. The scaling is tested for the diffusion g(t), the shear relaxation modulus G(t), and the Rouse mode autocorrelations of melts of C(120)H(242), C(400)H(802), and C(1000)H(2002). The derived dynamical scaling procedure is very useful to reduce run-time in mesoscale computer simulations, especially if pairwise friction is applied. PMID- 16375507 TI - Rapid shear viscosity calculation by momentum impulse relaxation molecular dynamics. AB - Recently, Arya et al. [J. Chem. Phys. 113, 2079 (2000)] introduced a new molecular dynamics method to rapidly compute the viscosity of fluids. The technique, termed momentum impulse relaxation (MIR), involves the imposition of a Gaussian velocity profile on an equilibrated system, after which the decay in the profile is monitored as a function of time. The shear viscosity is computed by matching the rate of decay of the velocity profile to the corresponding solution of the Navier-Stokes equation. The method was originally applied to simple systems (argon and n-butane) and found to give a comparable accuracy to conventional equilibrium and nonequilibrium methods with more than an order of magnitude reduction in computing time. In this work, we extend and generalize the method to examine larger molecules with higher viscosities than have been examined previously. A detailed analysis of the method is given, including the effect the velocity boundary conditions have on the viscosity, the sensitivity of the results to the velocity profile fitting procedure, the effect of preequilibration of the Gaussian profile, and the effect the system size and box shape have on the accuracy and speed of the method. It is shown that the MIR method can be extended to treat multiatom systems without loss of accuracy or computational efficiency. PMID- 16375508 TI - Thermal breaking of spanning water networks in the hydration shell of proteins. AB - The presence of a spanning hydrogen-bonded network of water at the surface of biomolecules is important for their conformational stability, dynamics, and function. We have studied by computer simulations the clustering and percolation of water in the hydration shell of a small elastinlike peptide (ELP) and the medium-size protein staphylococcal nuclease (SNase), in aqueous solution. We have found that in both systems a spanning network of hydration water exists at low temperatures and breaks up with increasing temperature via a quasi-two dimensional percolation transition. The thermal breaking of the spanning water network occurs at biologically relevant temperatures, in the temperature range, which is close to the temperature of the "inverse temperature transition" of ELP and the unfolding temperature of SNase, respectively. PMID- 16375509 TI - Structural thermodynamics of lamellar cationic lipid-DNA complex: DNA compressibility modulus. AB - We have studied theoretically the compressibility modulus B of DNA and complexation adsorption isotherms of DNA and lipids, as a function of DNA spacing d(DNA) and NaCl electrolyte concentration, respectively, in isoelectric states of lamellar DNA/cationic lipid (CL) self-assemblies. The electrostatic free energy derived from the Poisson-Boltzmann theory predicts partial agreement with measured B values for interhelical separations d(DNA)>33 A when use is made of a fit of hydration repulsion from bulk DNA hexagonal phases in solution. For lower interchain separations the prediction worsens due to the hydration interaction that overcomes the electrostatic contribution. An exact match of the system's counterion electrochemical potentials and the coions of salt in aqueous phase leads to the electrostatic part of the free energy that renders isotherms of d(DNA) versus ionic strength in qualitative consistency with general trends of available experimental data of CL-DNA complexes. PMID- 16375510 TI - Observation of fragile-to-strong liquid transition in surface water in CeO2. AB - A quasielastic neutron-scattering experiment carried out on a backscattering spectrometer with sub-micro eV resolution in the temperature range of 200-250 K has revealed the dynamics of surface water in cerium oxide on the time scale of hundreds of picoseconds. This slow dynamics is attributed to the translational mobility of the water molecules in contact with the surface hydroxyl groups. The relaxation function of this slow motion can be described by a slightly stretched exponential with the stretch factor exceeding 0.9, which indicates almost a Debye type dynamics. Down to about 220 K, the temperature dependence of the residence time for water molecules follows a Vogel-Fulcher-Tamman law with the glass transition temperature of 181 K. At lower temperatures, the residence time behavior abruptly changes, indicating a fragile-to-strong liquid transition in surface water at about 215 K. PMID- 16375511 TI - Energy band gaps and lattice parameters evaluated with the Heyd-Scuseria Ernzerhof screened hybrid functional. AB - This work assesses the Heyd-Scuseria-Ernzerhof (HSE) screened Coulomb hybrid density functional for the prediction of lattice constants and band gaps using a set of 40 simple and binary semiconductors. An extensive analysis of both basis set and relativistic effects is given. Results are compared with established pure density functionals. For lattice constants, HSE outperforms local spin-density approximation (LSDA) with a mean absolute error (MAE) of 0.037 A for HSE vs 0.047 A for LSDA. For this specific test set, all pure functionals tested produce MAEs for band gaps of 1.0-1.3 eV, consistent with the very well-known fact that pure functionals severely underestimate this property. On the other hand, HSE yields a MAE smaller than 0.3 eV. Importantly, HSE correctly predicts semiconducting behavior in systems where pure functionals erroneously predict a metal, such as, for instance, Ge. The short-range nature of the exchange integrals involved in HSE calculations makes their computation notably faster than regular hybrid functionals. The current results, paired with earlier work, suggest that HSE is a fast and accurate alternative to established density functionals, especially for solid state calculations. PMID- 16375512 TI - Density-functional calculations of relativistic spin-orbit effects on nuclear magnetic shielding in paramagnetic molecules. AB - Terms arising from the relativistic spin-orbit effect on both hyperfine and Zeeman interactions are introduced to density-functional theory calculation of nuclear magnetic shielding in paramagnetic molecules. The theory is a generalization of the former nonrelativistic formulation for doublet systems and is consistent to O(alpha4), the fourth power of the fine structure constant, for the spin-orbit terms. The new temperature-dependent terms arise from the deviation of the electronic g tensor from the free-electron g value as well as spin-orbit corrections to hyperfine coupling tensor A, the latter introduced in the present work. In particular, the new contributions include a redefined isotropic pseudocontact contribution that consists of effects due to both the g tensor and spin-orbit corrections to hyperfine coupling. The implementation of the spin-orbit terms makes use of all-electron atomic mean-field operators and/or spin-orbit pseudopotentials. Sample results are given for group-9 metallocenes and a nitroxide radical. The new O(alpha4) corrections are found significant for the metallocene systems while they obtain small values for the nitroxide radical. For the isotropic shifts, none of the three beyond-leading-order hyperfine contributions are negligible. PMID- 16375513 TI - Nucleation-controlled multiphase transitions. AB - Thermodynamics and kinetics of phase transitions in multiphase systems have been discussed. Thermodynamically admissible transitions have been identified and transition kinetics described in terms of the extended Kolmogoroff-Avrami-Evans model. Different combinations of transitions have been described as directed graphs. Graph nodes represented individual phases, graph edges--transitions. Superposition of parallel transitions in various mother phases, simultaneous transition of the same mother phase into different target phases, and successive (chain) transitions have been analyzed. Detailed solutions for a three-phase system consisting of one liquid phase and two polymorphic solid phases have been presented. PMID- 16375514 TI - Orbital-free embedding applied to the calculation of induced dipole moments in CO2...X (X = He, Ne, Ar, Kr, Xe, Hg) van der Waals complexes. AB - The orbital-free frozen-density embedding scheme within density-functional theory [T. A. Wesolowski and A. Warshel, J. Phys. Chem. 97, 8050 (1993)] is applied to the calculation of induced dipole moments of the van der Waals complexes CO2...X (X = He, Ne, Ar, Kr, Xe, Hg). The accuracy of the embedding scheme is investigated by comparing to the results of supermolecule Kohn-Sham density functional theory calculations. The influence of the basis set and the consequences of using orbital-dependent approximations to the exchange correlation potential in embedding calculations are examined. It is found that in supermolecular Kohn-Sham density-functional calculations, different common approximations to the exchange-correlation potential are not able to describe the induced dipole moments correctly and the reasons for this failure are analyzed. It is shown that the orbital-free embedding scheme is a useful tool for applying different approximations to the exchange-correlation potential in different subsystems and that a physically guided choice of approximations for the different subsystems improves the calculated dipole moments significantly. PMID- 16375515 TI - Calculation of specific, highly excited vibrational states based on a Davidson scheme: application to HFCO. AB - We present the efficiency of a new modified Davidson scheme which yields selectively one high-energy vibrationally excited eigenstate or a series of eigenstates. The calculation of a highly vibrationally excited state psi located in a dense part of the spectrum requires a specific prediagonalization step before the Davidson scheme. It consists in building a small active space P containing the zero-order states which are coupled with the zero-order description of the eigenstate of interest. We propose a general way to define this active space P which plays a crucial role in the method. The efficiency of the method is illustrated by computing and analyzing the high-energy excited overtones of the out-of-plane mode [formula: see text] in HFCO. These overtone energies correspond to the 234th, 713th, and 1774th energy levels in our reference basis set which contains roughly 140,000 states. One of the main advantages of this Davidson scheme comes from the fact that the eigenstate and eigenvalue convergence can be assessed during the iterations by looking at the residual [formula: see text]. The maximum value epsilon allowed for this residual constitutes a very sensitive and efficient parameter which sets the accuracy of the eigenvalues and eigenstates, even when the studied states are highly excited and are localized in a dense part of the spectrum. The physical analysis of the eigenstates associated with the 5th, 7th, and 9th out-of-plane overtones in HFCO provides some interesting information on the energy localization in this mode and on the role played by the in-plane modes. Also, it provides some ideas on the numerical methods which should be developed in the future to tackle higher-energy excited states in polyatomics. PMID- 16375516 TI - Coherent excitation of a two-state system by a linearly chirped Gaussian pulse. AB - This work presents an analytic description of coherent excitation of a two-state quantum system by an external field with a Gaussian temporal shape and a linear frequency sweep. A very accurate analytic approximation to the transition probability in terms of the Lambert function is derived by using the Dykhne-Davis Pechukas approach. This approximation provides analytic expressions for the frequency and the amplitude of the probability oscillations and for the ranges of interaction parameters where high transition probability is obtained. PMID- 16375517 TI - Transition probabilities of a string oscillator subject to impulsive collisions with a heavy mass point. AB - Impulsive linear collisions between a string oscillator (a one-dimensional particle in a box) and a mass point are studied quantum mechanically. In the limit of a very heavy mass point (which corresponds classically to many collisions during a single encounter) the transition probabilities are determined exactly. The result permits a discussion of the mixed quantum-classical regime where the collider becomes almost classical while the oscillator remains quantum mechanical. While the average transition probabilities P(m-->n) are well reproduced by the Ehrenfest mean-field approximation, the prediction for the superimposed high-frequency resonance structure is qualitatively wrong for a genuine quantum oscillator. Only if the oscillator is also almost classical and if (m-n)2 square root(mu) << m, where mu is the mass ratio collider/oscillator, this structure is correctly predicted by the Ehrenfest approximation. PMID- 16375518 TI - Semiclassical nonadiabatic dynamics using a mixed wave-function representation. AB - Nonadiabatic effects in quantum dynamics are described using a mixed polar/coordinate space representation of the wave function. The polar part evolves on dynamically determined potential surfaces that have diabatic and adiabatic potentials as limiting cases of weak localized and strong extended diabatic couplings. The coordinate space part, generalized to a matrix form, describes transitions between the surfaces. Choice of the effective potentials for the polar part and partitioning of the wave function enables one to represent the total wave function in terms of smooth components that can be accurately propagated semiclassically using the approximate quantum potential and small basis sets. Examples are given for two-state one-dimensional problems that model chemical reactions that demonstrate the capabilities of the method for various regimes of nonadiabatic dynamics. PMID- 16375519 TI - Pair potentials from diffraction data on liquids: a neural network solution. AB - The inverse theorem of liquids states a one to one correspondence between classical mechanical pair potentials and structural functions. Molecular-dynamics and Monte Carlo simulations provide exact structural functions for known pair interactions. There is no exact or widespread method in the opposite direction, where the pair interactions are to be determined from a priori known pair correlation functions or structure factors. The methods based on the integral equation theories of liquids are approximate and the iterative refinements of pair potentials with simulations take a long time. We applied artificial neural networks to get pair interactions from known structure factors in this study. We performed molecular-dynamics simulations on one-component systems with different pair potentials and the structure factors were calculated. To optimize (train) the weights of neural networks 2000 pair interaction-structure factor pairs were used. The performance of the method was tested on further 200 data pairs. The method provided reasonable potentials for the majority of the systems opening a "quick and dirty" method to determine pair interactions. PMID- 16375520 TI - Theory and method for calculating resonance Raman scattering from resonance polarizability derivatives. AB - We present a method to calculate both normal Raman-scattering (NRS) and resonance Raman-scattering (RRS) spectra from the geometrical derivatives of the frequency dependent polarizability. In the RRS case, the polarizability derivatives are calculated from resonance polarizabilities by including a finite lifetime of the electronic excited states using time-dependent density-functional theory. The method is a short-time approximation to the Kramers, Heisenberg, and Dirac formalism. It is similar to the simple excited-state gradient approximation method if only one electronic excited state is important, however, it is not restricted to only one electronic excited state. Since the method can be applied to both NRS and RRS, it can be used to obtain complete Raman excitation profiles. To test the method we present the results for the S2 state of uracil and the S4, S3, and S2 states of pyrene. As expected, the results are almost identical to the results obtained from the excited-state gradient approximation method. Comparing with the experimental results, we find in general quite good agreement which enables an assignment of the experimental bands to bands in the calculated spectrum. For uracil the inclusion of explicit waters in the calculations was found to be necessary to match the solution spectra. The calculated resonance enhancements are on the order of 10(4)-10(6), which is in agreement with experimental findings. For pyrene the method is also able to distinguish between the three different electronic states for which experimental data are available. The neglect of anharmonicity and solvent effects in the calculations leads to some discrepancy between theory and experiment. PMID- 16375521 TI - Information entropy, information distances, and complexity in atoms. AB - Shannon information entropies in position and momentum spaces and their sum are calculated as functions of Z(2 < or = Z < or = 54) in atoms. Roothaan-Hartree Fock electron wave functions are used. The universal property S = a + b ln Z is verified. In addition, we calculate the Kullback-Leibler relative entropy, the Jensen-Shannon divergence, Onicescu's information energy, and a complexity measure recently proposed. Shell effects at closed-shell atoms are observed. The complexity measure shows local minima at the closed-shell atoms indicating that for the above atoms complexity decreases with respect to neighboring atoms. It is seen that complexity fluctuates around an average value, indicating that the atom cannot grow in complexity as Z increases. Onicescu's information energy is correlated with the ionization potential. Kullback distance and Jensen-Shannon distance are employed to compare Roothaan-Hartree-Fock density distributions with other densities of previous works. PMID- 16375522 TI - Isomers of NCO2: IR-absorption spectra of ONCO in solid Ne. AB - Irradiation of a Ne matrix sample containing NO and CO near 4 K with an ArF excimer laser at 193 nm yielded new lines at 2045.1 and 968.0 cm(-1) that were depleted upon secondary photolysis at 308 nm. These lines are assigned to C=O stretching and mixed stretching modes of ONCO, based on results of 15N-, 13C-, and 18O-isotopic experiments and quantum-chemical calculations. These calculations using density-functional theory (B3LYP and PW91PW91/aug-cc-pVTZ) predict five stable isomers of NCO2: ONCO, NCOO, N-cyc-CO2, CNOO, and cyc-CNOO, listed in order of increasing energy. According to B3LYP calculations, ONCO has a trans configuration, with bond angles of angleONC approximately 136.3 degrees and angleNOC approximately 160.7 degrees. Calculated vibrational wave numbers, IR intensities, 15N-, 13C-, and 18O-isotopic shifts for ONCO agree satisfactorily with experimental results. ONCO was formed from reaction of CO with NO in its excited state. PMID- 16375523 TI - The vibrational energy pattern in acetylene VII: (12)C(13)CH2. AB - In (12)C(13)CH(2) 129 vibrational term values up to 10,000 cm(-1) are merged, about 60% of which are newly reported. They are fitted using an effective Hamiltonian with a standard deviation of 0.22 cm(-1). The vibrational assignments and vibrational constants are listed and discussed. The energy pattern is found to be very similar to the one in (12)C2H2 with additional anharmonic resonances arising from the lack of u/g character in the asymmetric isotopolog. PMID- 16375524 TI - The UV photodissociation dynamics of ClO radical using velocity map ion imaging. AB - We have studied the wavelength-dependent photodissociation dynamics of jet-cooled ClO radical from 235 to 291 nm using velocity map ion imaging. We find that Cl(2P(3/2))+O(1D(2)) is the dominant channel above the O(1D(2)) threshold with minor contributions from the Cl(2P(J))+O(3P(J)) and Cl(2P(1/2))+O(1D(2)) channels. We have measured the photofragment angular distributions for each dissociation channel and find that the A 2pi state reached via a parallel transition carries most of the oscillator strength above the O(1D(2)) threshold. The formation of O(3P(J)) fragments with positive anisotropy is evidence of curve crossing from the A 2pi state to one of several dissociative states. The curve crossing probability increases with wavelength in good agreement with previous theoretical calculations. We have directly determined the O(1D(2)) threshold to be 38,050+/-20 cm(-1) by measuring O(1D(2)) quantum yield in the wavelength range of 260-270 nm. We also report on the predissociation dynamics of ClO below the O(1D(2)) threshold. We find that the branching ratio of Cl(2P(3/2))/Cl(2P(1/2)) is 1.5+/-0.1 at both 266 and 291 nm. The rotational depolarization of the anisotropy parameters of the Cl(2P(3/2)) fragments provides predissociation lifetimes of 1.5+/-0.2 ps for the 9-0 band and 1.0+/-0.4 ps for the 8-0 band, in reasonable agreement with previous spectroscopic and theoretical studies. PMID- 16375525 TI - Ab initio relativistic calculation of the RbCs molecule. AB - We apply the relativistic configuration-interaction valence-bond method to calculate various characteristics of the alkali-metal RbCs dimer. These include the electronic potentials and transition dipole moments between the ground and first excited states and permanent dipole moments of the X 1sigma+ and a 3sigma+ states of the ground configuration. In addition, we estimate the lifetime of the rovibrational levels of the X state due to blackbody radiation. These data can help experimentalists to optimize photoassociative formation of ultracold RbCs molecules and their longevity in a trap or in an optical lattice. Extended basis sets, constructed from Dirac-Fock and Sturm's orbitals, have been used to ensure convergence of our calculations. We compare our data with other theoretical and experimental results when they were available. PMID- 16375526 TI - SiH2Cl2: ab initio anharmonic force field, dipole moments, and infrared vibrational transitions. AB - The vibrational spectra of SiH2Cl2 have been recorded in the 1000-13,000 cm(-1) region, utilizing the Fourier-transform spectroscopy and Fourier-transform intracavity laser absorption spectroscopy. Totally 61 band centers and intensities are derived from the infrared spectra. An ab initio quartic force field is obtained by applying the second-order Moller-Plesset perturbation theory and correlation-consistent polarized valence triplet-zeta basis sets [J. Chem. Phys. 90, 1007 (1989); 98, 1358 (1993)]. Most observed bands are assigned by the vibration analysis based on the second-order perturbation theory. Reduced dimensional ab initio dipole moment functions (two dimensional and three dimensional) have also been calculated to investigate the absolute band intensities of the SiH2 chromophore. The calculated values agree reasonably with the observed ones. PMID- 16375527 TI - State-resolved collisional quenching of vibrationally excited pyrazine (E(vib) = 37,900 cm(-1)) by D35Cl(v = 0). AB - Supercollision relaxation of highly vibrationally excited pyrazine (E(vib) = 37,900 cm(-1)) with D35Cl is investigated using high-resolution transient IR diode laser absorption spectroscopy at 4.4 microm. Highly excited pyrazine is prepared by pulsed UV excitation at 266 nm, followed by rapid radiationless decay to the ground electronic state. The rotational energy distribution of the scattered DCl (v = 0,J) molecules with J = 15-21 is characterized by T(rot) = 755+/-90 K. The relative translational energy increases as a function of rotational quantum number for DCl with T(rel) = 710+/-190 K for J = 15 and T(rel) = 1270+/-240 K for J = 21. The average change in recoil velocity correlates with the change in rotational angular momentum quantum number and highlights the role of angular momentum in energy gain partitioning. The integrated energy-transfer rate for appearance of DCl (v = 0,J = 15-21) is k(2)(int) = 7.1x10(-11) cm3 molecule(-1) s(-1), approximately one-eighth the Lennard-Jones collision rate. The results are compared to earlier energy gain measurements of CO2 and H2O. PMID- 16375528 TI - Lifetime and yield of metastable Ar2(+) ions. AB - Ar2(+) ions produced in a cooled supersonic expansion by electron-impact ionization are accelerated at 2.5 keV and kept during few milliseconds inside a linear electrostatic trap. The lifetime of the metastable Ar2(+) ion is determined from the measurement of the rate of the argon atoms escaping the trap. The lifetime and the relative metastable populations are measured as a function of the pressure and temperature in the supersonic expansion, i.e., of the mean cluster size. Possible mechanisms responsible for the metastable formation are discussed. PMID- 16375529 TI - Three-isotope plot of fractionation in photolysis: a perturbation theoretical expression. AB - The slope of the three-isotope plot for the isotopomer fractionation by direct or nearly direct photodissociation is obtained using a perturbation theoretical analysis. This result, correct to first order in the mass difference, is the same as that for equilibrium chemical exchange reactions, a similarity unexpected a priori. A comparison is made with computational results for N2O photodissociation. This theoretical slope for mass-dependent photolytic fractionation can be used to analyze the data for isotopic anomalies in spin allowed photodissociation reactions. Earlier work on chemical equilibria is extended by avoiding a high-temperature approximation. PMID- 16375530 TI - Rare-gas insertion compounds of perfluorobenzene: aromaticity of some unstable species. AB - Calculations on the novel argon insertion compounds C6F6Ar(n), n = 1-6, where the argon atoms are inserted into the C-F bonds in perfluorobenzene, suggest that all possible species, with one to six inserted argon atoms, occupy minima on their respective potential energy surfaces. Ring-current plots using the ipsocentric model indicate that there is no disruption of the aromatic pi system upon argon insertion, and consequently all insertion compounds are aromatic according to the magnetic criterion. The barrier height for decomposition of the single-insertion compound, C6F6Ar, into C6F6+Ar is 19.5 kcal/mol at HF6-311G** and 29.5 kcal/mol at B3LYP6-311G**, suggesting that, although clearly thermodynamically unstable, argon-perfluorobenzene insertion compounds may be stable kinetically. Preliminary calculations indicate that other rare gas-perfluorobenzene insertion compounds may also be metastable. Both C6F6Ne and C6F6He are predicted to occupy minima on their respective potential energy surfaces. PMID- 16375531 TI - Mass-analyzed threshold ionization study of vinyl bromide cation in the first excited electronic state using vacuum-ultraviolet radiation generated by four wave mixing in Hg. AB - The vibrational spectrum of the vinyl bromide cation in the first excited electronic state A 2A' was obtained by one-photon mass-analyzed threshold ionization (MATI) spectroscopy. The use of an improved vacuum-ultraviolet radiation source based on four-wave sum frequency mixing in Hg resulted in excellent sensitivity for MATI signals. From the MATI spectrum, the ionization energy to the A 2A' state of the cation was determined to be 10.9150+/-0.0006 eV. Nearly complete vibrational assignments for the MATI peaks were possible by utilizing the vibrational frequencies and Franck-Condon factors calculated at the density-functional theory (DFT) and time-dependent DFT/B3LYP levels with the 6 311+G(df,p) basis set. PMID- 16375532 TI - Rotational excitation cross sections of para-H2 + para-H2 collisions. A full dimensional wave-packet propagation study using an exact form of the kinetic energy. AB - A full-dimensional quantum dynamical study of the rotational excitation in para para H2 + H2 collisions using the potential-energy surface of Boothroyd et al. [J. Chem. Phys. 116, 666 (2002)] is reported. The multiconfiguration time dependent Hartree algorithm is utilized to propagate wave packets and the cross sections for collision energies up to 1.2 eV are determined by a flux analysis through the interaction of the wave packet with a complex absorbing potential. Calculations for a collection of total angular momenta up to J = 70 are performed; the missing channels are obtained with a J-interpolation algorithm. PMID- 16375533 TI - Cross sections of the O+ + H2 --> OH+ + H ion-molecule reaction and isotopic variants (D2, HD): quasiclassical trajectory study and comparison with experiments. AB - A dynamics study [cross section and microscopic mechanism versus collision energy (E(T))] of the reaction O+ + H2 --> OH+ + H, which plays an important role in Earth's ionosphere and interstellar chemistry, was conducted using the quasiclassical trajectory method, employing an analytical potential energy surface (PES) recently derived by our group [R. Martinez et al., J. Chem. Phys. 120, 4705 (2004)]. Experimental excitation functions for the title reaction, as well as its isotopic variants with D2 and HD, were near-quantitatively reproduced in the calculations in the very broad collision energy range explored (E(T) = 0.01-6.0 eV). Intramolecular and intermolecular isotopic effects were also examined, yielding data in good agreement with experimental results. The reaction occurs via two microscopic mechanisms (direct and nondirect abstraction). The results were satisfactorily interpreted based on the reaction probability and the maximum impact parameter dependences with E(T), and considering the influence of the collinear [OHH]+ absolute minimum of the PES on the evolution from reactants to products. The agreement between theory and experiment suggests that the reaction mainly occurs through the lowest energy PES and nonadiabatic processes are not very important in the wide collision energy range analyzed. Hence, the PES used to describe this reaction is suitable for both kinetics and dynamics studies. PMID- 16375534 TI - A diffusion quantum Monte Carlo study on the lowest singlet and triplet electronic states of BN molecule. AB - Ab initio calculation of both the lowest singlet and triplet electronic states of BN has been performed by the fixed-node Ornstein-Uhlenbeck diffusion quantum Monte Carlo method with the floating spherical Gaussian orbitals and spherical Gaussian geminals. The Monte Carlo calculation gives equilibrium bond lengths and equilibrium harmonic frequencies of 1.3317(7) A and 1529(7) cm(-1), respectively, for the lowest triplet state and 1.2751(7) A and 1709(8) cm(-1), respectively, for the lowest singlet state. Also, the Monte Carlo calculation reports an energy separation of 178(83) cm(-1) between the two electronic states and recommends the ground state is the lowest triplet state. PMID- 16375535 TI - Weak metal-metal bonding in small manganese cluster ions, Mn(N)+ (N < or = 7). AB - The binding energies of manganese cluster ions Mn(N)+ (N = 5-7) were determined by the photodissociation experiments in the near-infrared and visible-photon energy ranges. The bond dissociation energies of Mn(N)+, D0(Mn(N-1)+...Mn), were obtained to be 1.70+/-0.08, 1.04+/-0.10, and 1.46+/-0.11 eV, respectively, for N = 5, 6, and 7 from the threshold energies for the two-atom loss processes and the bond dissociation energies of Mn3(+) and Mn4(+) reported previously [A. Terasaki et al., J. Chem. Phys. 117, 7520 (2002)]. Correspondingly, binding energies per atom are obtained to be 0.99+/-0.03, 1.00+/-0.03, and 1.06+/-0.03 eV/at. for N = 5, 6, and 7, respectively. A gradual increase in the binding energy from N = 2 to N = 7 shows an increasing contribution of nonbonding 3d orbitals to the bonding via weak hybridization with valence 4s orbitals as the cluster size increases. These binding energies per atom are still much smaller than the bulk cohesive energy of manganese (2.92 eV/at.), and this finding indicates exceptionally weak metal-metal bonds in this size range. PMID- 16375536 TI - Vibrational energy transfer in N2-N2 collisions: a new semiclassical study. AB - The vibrational energy relaxation in collisions between N2 molecules in the low- and medium-lying vibrationally excited levels was revisited using the semiclassical coupled-state method and the use of two different potential-energy surfaces having the same short-range potential recently determined from ab initio calculations but with different long-range interactions. Compared to the data reported in the classical work by Billing and Fisher [Chem. Phys. 43, 395 (1979)], the newly calculated vibration-to-translation rate constant K(1,0 / 0,0) is in much better agreement with the available experimental data over a large temperature interval, from T = 200 K up to T = 6000 K. Nevertheless, as far as the vibration-to-translation exchanges are concerned, the lower-temperature regime remains quite critical in that the new rate constants do not completely account for the rate constant curvature suggested by the experiments for temperatures lower than T = 500 K. The dependence of the state-selected vibration to-vibration rate constants, K(v,v-delta v / 0,1), both upon the vibrational quantum number v and the gas temperature are calculated. The substantial deviations from previously found behaviors could have major consequences for the vibrational kinetic modeling of N2-containing gas mixtures. PMID- 16375537 TI - A 193 nm laser photofragmentation time-of-flight mass spectrometric study of chloroiodomethane. AB - The photodissociation dynamics of chloroiodomethane (CH2ICl) at 193 nm has been investigated by employing the photofragment time-of-flight (TOF) mass spectrometric method. Using tunable vacuum ultraviolet undulator synchrotron radiation for photoionization sampling of nascent photofragments, we have identified four primary dissociation product channels: CH2Cl + I(2P(1/2))/I(2P(3/2)), CH2I + Cl(2P(1/2))/Cl(2P(3/2)), CHI + HCl, and CH2 + ICl. The state-selective detection of I(2P(3/2)) and I(2P(1/2)) has allowed the estimation of the branching ratio for I(2P(1/2)):I(2P(3/2)) to be 0.73:0.27. Theoretical calculations based on the time-dependent density-functional theory have been also made to investigate excited electronic potential-energy surfaces, plausible intermediates, and transition structures involved in these photodissociation reactions. The translation energy distributions derived from the TOF measurements suggest that at least two dissociation mechanisms are operative for these photodissociation processes. One involves the direct dissociation from the 2 1A' state initially formed by 193 nm excitation, leading to significant kinetic-energy releases. For the I-atom and Cl-atom elimination channels, the fragment kinetic-energy releases observed via this direct dissociation mechanism are consistent with those predicted by the impulsive dissociation models. Other mechanisms are likely predissociative or statistical in nature from the lower 1 1A' and 1 1A'' states and/or the ground X 1A' state populated by internal conversion from the 2 1A' state. On the basis of the maximum kinetic-energy release for the formation of CH2Cl + I(2P(1/2)), we have obtained a value of 53+/-2 kcal/mol for the 0 K bond dissociation energy of I CH2Cl. The intermediates and transition structures for the CHI + HCl and CH2 + ICl product channels have been also investigated by ab initio quantum calculations at the MP2(full)/6-311G(d) and B3LYP(full)/6-11G(d) levels of theory. The maximum kinetic-energy releases observed for the CHI + HCl and CH2 + ICl channels are consistent with the interpretation that the formation of CHI and CH2 in their ground triplet states is not favored. PMID- 16375538 TI - Are azobenzenophanes rotation-restricted? AB - We simulated the photoisomerization dynamics of an azobenzenophane with a semiclassical surface hopping approach and a semiempirical reparametrized quantum mechanics/molecular mechanics Hamiltonian. Only one of the two azobenzene chromophores in the molecule is taken into account quantum mechanically: the other one is treated by molecular mechanics. Both n-->pi* and pi-->pi* excitations are considered. Our results show that the photoisomerization reaction mainly involves the rotation around the N=N double bond. The excited state relaxation features are in qualitative agreement with experimental time-resolved fluorescence results. PMID- 16375539 TI - Pressure broadening and fine-structure-dependent predissociation in oxygen B 3sigma(u)-, v = 0. AB - Both laser-induced fluorescence and cavity ring-down spectral observations were made in the Schumann-Runge band system of oxygen, using a novel-type ultranarrow deep-UV pulsed laser source. From measurements on the very weak (0,0) band pressure broadening, pressure shift, and predissociation line-broadening parameters were determined for the B 3sigma(u)-, v = 0,F(i) fine-structure components for various rotational levels in O2. The information content from these studies was combined with that of entirely independent measurements probing the much stronger (0,10), (0,19), and (0,20) Schumann-Runge bands involving preparation of vibrationally excited O2 molecules via photolysis of ozone. The investigations result in a consistent set of predissociation widths for the B 3sigma(u)-, v = 0 state of oxygen. PMID- 16375540 TI - Structure and conformation properties of 1-alkyl-3-methylimidazolium halide ionic liquids: a density-functional theory study. AB - The structures and conformational properties of 1-alkyl-3-methylimidazolium halide ionic liquids have been studied with a Becke's 3 Parameter functional method. The interaction mechanisms between the cation and the anion in 1-ethyl-3 methylimidazolium (Emim+) halide and 1-butyl-3-methylimidazolium (Bmim+) halide ionic liquids were investigated using 6-31G*, 6-31++G**, and 6-311++G** basis sets. Forty structures of different ion pairs were optimized and geometrical parameters of them have been discussed in details. Halide ions (Cl- or Br-) have been gradually placed in different regions around imidazolium cation and the interaction energies between the anion and the cation have been calculated. Theoretical results indicate that there are four activity regions in the vicinity of the imidazolium cations, in these regions the imidazolium cations and the halide anions formed stable ion pairs. Imidazolium cations can form hydrogen bond interactions with one, two or three but no more than three nearest halide anions. The halide ions are situated in hydrogen bond positions rather than at random. PMID- 16375541 TI - Anomalous dielectric relaxation in strong ac external fields. AB - Dielectric relaxation of complex polar fluids is considered in the context of the anomalous diffusion characterized by a fractional parameter alpha < or = 1 (subdiffusion). An infinite hierarchy of three-term differential-recurrence equations governing the time evolution of the electric polarization is established by following a purely phenomenological procedure. The matrix continued fraction method is used to derive the exact numerical solution of the stationary regime for an assembly of nonelectrically interacting, polar symmetric top molecules in presence of a strong ac electric field. The results so obtained are valid to any order in the field strength parameter gamma1, thus extending previous theories applicable to fields of very small amplitudes only. This is illustrated by Cole-Cole diagrams and three-dimensional relaxation spectra for the first- and third-harmonic components of the electric polarization as a function of alpha, gamma1, and the angular frequency. PMID- 16375542 TI - Thermal conductivity of ethanol. AB - We present a factorization of the Ewald sum permitting efficient computation of the reciprocal space part of the molecular representation for the heat flux vector. We use the derived expression to evaluate thermal conductivity of a model of ethanol at several near-ambient state points. PMID- 16375543 TI - Do organic solutes experience specific interactions with ionic liquids? AB - In an attempt to understand the nature of interactions between organic solutes and room temperature ionic liquids, temperature-dependent rotational relaxation of two structurally similar nondipolar solutes--2,5-dimethyl-1,4-dioxo-3,6 diphenylpyrrolo[3,4-c]pyrrole (DMDPP) and 1,4-dioxo-3,6-diphenylpyrrolo[3,4 c]pyrrole (DPP)--has been examined in 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim+][PF6(-)]). Even with the ionic liquid, where the cation and the anion are strongly associated, the solute DPP experiences specific interactions, which is evident from its reorientation times that are 50%-60% longer in relation to DMDPP. It has been noticed that the reorientation times of both the solutes are faster in [bmim+][PF6(-)] than in glycerol, which is also a strongly associated solvent and whose viscosity is similar to the ionic liquid. This observation has been explained by taking into consideration the relative sizes of the solvents. By comparing the ratios of the reorientation times of DPP to DMDPP, in [bmim+][PF6(-)] and glycerol, it has been deduced that the strengths of the interaction between DPP-[bmim+][PF6(-)] and DPP-glycerol are the same. PMID- 16375544 TI - Direct determination of phase behavior of square-well fluids. AB - We have combined Gibbs ensemble Monte Carlo simulations with the aggregation volume-biased method in conjunction with the Gibbs-Duhem method to provide the first direct estimates for the vapor-solid, vapor-liquid, and liquid-solid phase coexistences of square-well fluids with three different ranges of attraction. Our results are consistent with the previous simulations and verify the notion that the vapor-liquid coexistence behavior becomes metastable for cases where the attraction well becomes smaller than 1.25 times the particle diameter. In these cases no triple point is found. PMID- 16375545 TI - Spiral pattern in chlorite-iodide-malonic acid reaction: a theoretical and numerical study. AB - The development of spiral pattern in a model representing chlorite-iodide-malonic acid reaction is investigated theoretically and numerically. We have carried out a multiple scale analysis of the model to identify the experimentally admissible parameter range and the appropriate perturbation for shifting Hopf bifurcation boundary towards the oscillating region. Our theoretical analysis is corroborated by numerical simulation. PMID- 16375546 TI - Shear stress relaxation and physical aging study on simple glass-forming materials. AB - Relaxation and aging behaviors in three supercooled liquids: m-toluidine, glycerol, and sucrose benzoate have been studied by shear stress relaxation experiments in the time domain above and below their nominal glass transition temperatures. For the equilibrium state, the current study provides new data on the behavior of organic complex fluids. The shape of the relaxation function as characterized by the stretching exponent beta is discussed considering that a time-temperature master curve can be constructed even though the beta's for the individual response curves at each temperature vary systematically. In the nonequilibrium state, isothermal physical aging experiments at different glassy structures reveal that the effect of the aging process on the mechanical shear relaxation in these simple glass formers is similar to that observed in polymeric and other systems. Departure from the Vogel-Fulcher-Tamman behavior after the samples have aged back to equilibrium in the glassy state is observed for m toluidine and, less strongly, for glycerol but not for sucrose benzoate. An inherent structure-based energy landscape concept is briefly discussed to account for the slow dynamics during the physical aging process. PMID- 16375547 TI - Structure of highly asymmetric hard-sphere mixtures: an efficient closure of the Ornstein-Zernike equations. AB - A simple modification of the reference hypernetted chain (RHNC) closure of the multicomponent Ornstein-Zernike equations with bridge functions taken from Rosenfeld's hard-sphere bridge functional is proposed. Its main effect is to remedy the major limitation of the RHNC closure in the case of highly asymmetric mixtures--the wide domain of packing fractions in which it has no solution. The modified closure is also much faster, while being of similar complexity. This is achieved with a limited loss of accuracy, mainly for the contact value of the big sphere correlation functions. Comparison with simulation shows that inside the RHNC no-solution domain, it provides a good description of the structure, while being clearly superior to all the other closures used so far to study highly asymmetric mixtures. The generic nature of this closure and its good accuracy combined with a reduced no-solution domain open up the possibility to study the phase diagram of complex fluids beyond the hard-sphere model. PMID- 16375548 TI - Influence of an electric field on the buoyancy-driven instabilities. AB - The influence of dc electric fields (EFs) on the development of buoyancy-driven instabilities of reaction fronts is investigated experimentally in a modified Hele-Shaw cell for the arsenous acid-iodate system. Assessment of effects of external EFs is made both visually and through dispersion curves. It is shown that density fingering, observed on ascending fronts, is suppressed by the EF if the front propagates towards the positive electrode and is enhanced when the front propagates towards the negative electrode. The stabilizing (destabilizing) effects include slower (faster) development of fingers and the decrease (increase) in their numbers. The descending front, stable under no EF conditions, remains stable when an EF is applied with the positive electrode facing the approaching front. When the descending front faces the negative electrode, the tiny fingerlike structure develops after quite a long time. PMID- 16375549 TI - Mode coupling behavior of a Lennard-Jones binary mixture: a comparison between bulk and confined phases. AB - We present a quantitative comparison at equivalent thermodynamical conditions of bulk and confined dynamical properties of a Lennard-Jones binary mixture upon supercooling. Both systems had been previously found to display a behavior in agreement with the mode coupling theory of the evolution of glassy dynamics. Differences and analogies of behavior are discussed focusing, in particular, on the role of hopping in reducing spatially correlated dynamics in the confined system with respect to the bulk. PMID- 16375550 TI - High pressure-temperature Brillouin study of liquid water: evidence of the structural transition from low-density water to high-density water. AB - The structural transformations occurring to water from low-density (LDW) to high density (HDW) regimes have been studied by Brillouin scattering for the first time at temperatures up to 453 K and at pressures up to the solidification point. At ambient temperature (293 K) a discontinuity in pressure response of the sound velocity is observed. Furthermore, there are evident breaks in the linear behavior of log10 C11 versus log10(rho/rho0) when pressure increases up to 0.29, 0.21, and 0.19 GPa at the temperature of 293, 316, and 353 K, respectively. It is supposed to indicate the structural transition from LDW to HDW, and the possible transition boundary between LDW and HDW is in good agreement with the molecular dynamics simulation. PMID- 16375551 TI - The structure and crystallization of thin water films on Pt(111). AB - When water is adsorbed on Pt(111) above 135 K several different ice structures crystallize, depending on the thickness of the ice layer. At low coverage water forms extended islands of ice with a (square root(37) x square root(37))R25(o) unit cell, which compresses as the monolayer saturates to form a (square root(39) x square root(39))R16(o) structure. The square root(39) low-energy electron diffraction (LEED) pattern becomes more intense as the second layer grows, remaining bright for films up of 10-15 layers and then fading and disappearing for films more than ca. 40 layers thick. The ice multilayer consists of an ordered square root(39) wetting layer, on which ice grows as a crystalline film which progressively loses its registry to the wetting layer. Ice films more than ca. 50 layers thick develop a hexagonal LEED pattern, the entire film and wetting layer reorienting to form an incommensurate bulk ice. These changes are reflected in the vibrational spectra which show changes in line shape and intensity associated with the different ice structures. Thin amorphous solid water films crystallize to form the same phases observed during growth, implying that these structures are thermodynamically stable and not kinetic phases formed during growth. The change from a square root(39) registry to incommensurate bulk ice at ca. 50 layers is associated with a change in crystallization kinetics from nucleation at the Pt(111) interface in thin films to nucleation of incommensurate bulk ice in amorphous solid water films more than 50 layers thick. PMID- 16375552 TI - Electrolytes at spherical dielectric interfaces. AB - A variational theory is developed and applied to study the properties of dielectric spheres immersed in a symmetric electrolyte solution. In the limit that the radius of the sphere becomes much larger than the Debye screening length, the system reduces to that of a planar dielectric interface. For this case, the excess surface tension obtained by the variational theory reduces to the Onsager-Samaras [J. Chem. Phys. 2, 528 (1934)] limiting law at low electrolyte concentrations. As the radius of the dielectric sphere decreases, the excess surface tension also decreases. The implications of this work to protein salt interactions and the salting out of proteins are discussed. PMID- 16375553 TI - Ab initio surface reaction energetics of SiH4 and Si2H6 on Si(001)-(2 x 2). AB - First-principles pseudopotential calculations, within a simple dynamically constrained scheme, have been performed to investigate the reaction of 0.25 ML coverage of SiH4 and Si2H6 with the Si(001)-(2 x 2) surface. The silane molecule (SiH4) is adsorbed on to the surface at a number of different sites (on dimer, interrow, or intrarow) with varying barrier heights. Two distinct structures, which are similar in energy, arise from the initial dissociative reaction SiH4- >SiH3(silyl) + H, where the dissociated species are adsorbed either on the same dimer components or on adjacent dimer components. Several further decays of silyl from SiH4 are presented in two separate regimes of high and low ambient hydrogen coverages. The decomposition of silyl can form two different bridging structures: an on top or an intrarow bridging structure in both of the two hydrogen coverage regimes. The disilane molecule (Si2H6) is also adsorbed upon this surface with varying energy barriers, resulting in a dissociation reaction where two SiH3 species are adsorbed on one dimer or in an adjacent dimer configuration. Plausible energy reaction paths for the above models are presented. The stability of the SiH2 species is also discussed. PMID- 16375554 TI - Fluorescence intermittency in blinking quantum dots: renewal or slow modulation? AB - We study the time series produced by blinking quantum dots, by means of an aging experiment, and we examine the results of this experiment in the light of two distinct approaches to complexity, renewal and slow modulation. We find that the renewal approach fits the result of the aging experiment, while the slow modulation perspective does not. We make also an attempt at establishing the existence of an intermediate condition. PMID- 16375555 TI - Ab initio density-functional study of the bridging addition of acrylonitrile on the Si(100) surface. AB - Acrylonitrile may react with the Si(100) surface in a number of ways. Fifteen different configurations have been identified. This study which complements an earlier study devoted to cycloadditions and bonds involving a single atom deals essentially with configurations involving cumulative double bonds. Factors affecting the relative stability of various mesomeric forms are shown to be the proximity to tetrahedral geometry and the compatibility of the dipole form with buckled structures. Tripods are shown to be a way to accommodate a priori unfavorable structures. PMID- 16375556 TI - The application of infrared spectroscopy to probe the surface morphology of alumina-supported palladium catalysts. AB - Five alumina-supported palladium catalysts have been prepared from a range of precursor compounds [palladium(II) nitrate, palladium(II) chloride, palladium(II) acetylacetonate, and tetraamminepalladium(II) tetraazidopalladate(II)] and at different metal loadings (1-7.3 wt %). Collectively, this series of catalysts provides a range of metal particle sizes (1.2-8.5 nm) that emphasize different morphological aspects of the palladium crystallites. The infrared spectra of chemisorbed CO applied under pulse-flow conditions reveal distinct groupings between metal crystallites dominated by low index planes and those that feature predominantly corner/edge atoms. Temperature-programmed infrared spectroscopy establishes that the linear CO band can be resolved into contributions from corner atoms and a combination of (111)(111) and (111)(100) particle edges. Propene hydrogenation has been used as a preliminary assessment of catalytic performance for the 1 wt % loaded catalysts, with the relative inactivity of the catalyst prepared from palladium(II) chloride attributed to a diminished hydrogen supply due to decoration of edge sites by chlorine originating from the preparative process. It is anticipated that refinements linking the vibrational spectrum of a probe molecule with surface structure and accessible adsorption sites for such a versatile catalytic substrate provide a platform against which structure/reactivity relationships can be usefully developed. PMID- 16375557 TI - Effects of frustration, confinement, and surface interactions on the dimerization of an off-lattice beta-barrel protein. AB - We study the effects of confinement, sequence frustration, and surface interactions on the thermodynamics of dimerization of an off-lattice minimalist beta-barrel protein using replica exchange molecular dynamics. We vary the degree of frustration of the protein by tuning the specificity of the hydrophobic interactions and investigate dimerization in confining spheres of different radii. We also investigate surface effects by tethering the first residue of one of the proteins to a uniformly repulsive surface. We find that increasing the confinement and frustration stabilize the dimer, while adding a repulsive surface decreases its stability. Different ensembles of structures, including properly dimerized and various partially dimerized states, are observed at the association transition temperature T(a), depending on the amount of frustration and whether a surface is present. The presence of a surface is predicted to alter the morphology of larger aggregates formed from partially unfolded dimeric conformations. PMID- 16375558 TI - Stability of Ca-montmorillonite hydrates: a computer simulation study. AB - Classic simulations are used to study interlayer structure, swelling curves, and stability of Ca-montmorillonite hydrates. For this purpose, NP(zz)T and muP(zz)T ensembles are sampled for ground level and given burial conditions. For ground level conditions, a double layer hydrate having 15.0 A of basal spacing is the predominant state for relative vapor pressures (p/p0) ranging 0.6-1.0. A triple hydrate counting on 17.9 A of interlaminar distance was also found stable for p/p0 = 1.0. For low vapor pressures, the system may produce a less hydrated but still double layer state with 13.5 A or even a single layer hydrate with 12.2 A of interlaminar distance. This depends on the established initial conditions. On the other hand, the effect of burial conditions is two sided. It was found that it enhances dehydration for all vapor pressures except for saturation, where swelling is promoted. PMID- 16375559 TI - Energy transfer in near-field optics. AB - When the probe tip of a near-field optical microscope illuminates nanoparticles with marked absorption bands, a large number of photons are absorbed before reaching the detector. These energy losses enhance the dark contrast usually observed in the vicinity of metallic nanoparticles. We demonstrate theoretically that this phenomenon can be exploited to image, in the optical frequency range, dissipative domains with a nanometer scale resolution. Simulations performed with noble-metal particles indicate that the detected signal significantly drops down when the excitation frequency is approaching the plasmon resonance of the particles. PMID- 16375560 TI - Upconversion luminescence, intensity saturation effect, and thermal effect in Gd2O3:Er3,Yb3+ nanowires. AB - In this paper, the upconversion luminescent properties of Gd2O3:Er3+,Yb3+ nanowires as a function of Yb concentration and excitation power were studied under 978-nm excitation. The results indicated that the relative intensity of the red emission (4F(9/2)-4I(15/2)) increased with increasing the Yb3+ concentration, while that of the green emission (4S(3/2)/2H(11/2)-4I(15/2)) decreased. As a function of excitation power in ln-ln plot, the green emission of 4S(3/2) 4I(15/2) yielded a slope of approximately 2, while the red emission of 4F(9/2) 4I(15/2) yielded a slope of approximately 1. Moreover, the slope decreased with increasing the Yb3+ concentration. This was well explained by the expanded theory of competition between linear decay and upconversion processes for the depletion of the intermediate excited states. As the excitation power density was high enough, the emission intensity of upconversion decreased due to thermal quenching. The thermal effect caused by the exposure of the 978-nm laser was studied according to the intensity ratio of 2H(11/2)-4I(15/2) to 4S(3/2) 4I(15/2). The practical sample temperature at the exposed spot as a function of excitation power and Yb3+ concentration was deduced. The result indicated that at the irradiated spot (0.5 x 0.5 mm2) the practical temperature considerably increased. PMID- 16375561 TI - Mechanism of adsorption in cylindrical nanopores: the roles of fluctuations and correlations in stabilizing the adsorbed phase. AB - The mechanism of the adsorption in nanometric cylindrical pores has been studied using grand canonical Monte Carlo simulations. The results have been analyzed from the point of view of microscopic correlations. It has been shown that the correlations between the energy components and between the energy and the number of adsorbed particles provide crucial information concerning the microscopic mechanism of the formation of adsorbed layers. Typical susceptibility functions have been calculated. They give the relations between the statistical correlations and the stability of the adsorbed system in different stages of adsorption. The numerical calculations have been carried out for Kr atoms adsorbed in an MCM-41 model porous material with pores of diameter d = 4 nm. The smooth-wall model as well as the model of a wall with micropores have been discussed. PMID- 16375562 TI - Liquid-vapor interfacial properties of water-ammonia mixtures: dependence on ammonia concentration. AB - The equilibrium and dynamical properties of the liquid-vapor interfaces of water ammonia mixtures are investigated by means of molecular-dynamics simulations. Altogether, we have simulated seven different systems of different concentration of ammonia. The inhomogeneous density, anisotropic orientational profiles, surface tension, and the pattern of hydrogen bonding are calculated for both water and ammonia molecules in order to characterize the location, width, thermodynamic aspects, and microscopic structure of the liquid-vapor interfaces of each of the water-ammonia systems. The dynamical aspects of the interfaces are investigated in terms of the anisotropic diffusion and dipole orientational relaxation of water and ammonia molecules. The properties of the interfaces are compared with those of the corresponding bulk phases. The present theoretical results are also compared with experimental findings wherever available. PMID- 16375563 TI - Continuum formulation of the Scheutjens-Fleer lattice statistical theory for homopolymer adsorption from solution. AB - Homopolymer adsorption from a dilute solution on an interacting (attractive) surface under static equilibrium conditions is studied in the framework of a Hamiltonian model. The model makes use of the density of chain ends n(1,e) and utilizes the concept of the propagator G describing conformational probabilities to locally define the polymer segment density or volume fraction phi; both n(1,e) and phi enter into the expression for the system free energy. The propagator G obeys the Edwards diffusion equation for walks in a self-consistent potential field. The equilibrium distribution of chain ends and, consequently, of chain conformational probabilities is found by minimizing the system free energy. This results in a set of model equations that constitute the exact continuum-space analog of the Scheutjens-Fleer (SF) lattice statistical theory for the adsorption of interacting chains. Since for distances too close to the surface the continuum formulation breaks down, the continuum model is here employed to describe the probability of chain configurations only for distances z greater than 2l, where l denotes the segment length, from the surface; instead, for distances z < or = 2l, the SF lattice model is utilized. Through this novel formulation, the lattice solution at z = 2l provides the boundary condition for the continuum model. The resulting hybrid (lattice for distances z < or = 2l, continuum for distances z > 2l) model is solved numerically through an efficient implementation of the pseudospectral collocation method. Representative results obtained with the new model and a direct application of the SF lattice model are extensively compared with each other and, in all cases studied, are found to be practically identical. PMID- 16375564 TI - Microscopic investigations of homogeneous nucleation in charged sphere suspensions. AB - We studied the homogeneous nucleation kinetics of an aqueous suspension of charged colloidal spheres under de-ionized conditions. Samples of equilibrium crystalline structure were shear molten and the metastable melt left to solidify after cessation of shear. At low particle number densities n, corresponding to low metastability of the melt, nucleation was monitored directly via video microscopy. We determined the nucleation rates gamma(t) by counting the number of newly appearing crystals in the observation volume per unit time. Using a suitable discrete adaptation of Avrami's [J. Chem. Phys. 7, 1003 (1939); ibid.8, 212 (1940); ibid.9, 177 (1941)] model for solidification via homogeneous nucleation and subsequent growth, we calculate the remaining free volume VF(t) to obtain the rate densities J(t) = gamma(t)/VF(t). We observe J(t) to rise steeply, display a plateau at a maximum rate density Jmax, and to decrease again. With increased n the plateau duration shrinks while Jmax increases. At low to moderate number densities fully solidified samples were analyzed by microscopy to obtain the grain-size distribution and the average crystallite size angle brackets(L). Under the assumption of stationarity, we obtained the nucleation rate density J(Avr), which increased strongly with increasing n. Interestingly, J(Avr) agrees quantitatively to Jmax and to J(Avr) as obtained previously from scattering data taken on the same sample at large n. Thus, by combination of different methods, reliable nucleation rate densities are now available over roughly one order of magnitude in n and eight orders of magnitude in J. PMID- 16375565 TI - A computer simulation study of the segregation of amphiphiles in binary immiscible matrices: short asymmetric copolymers in short homopolymers. AB - The bulk and interfacial properties of ternary mixtures with asymmetric amphiphiles (A2B8) in A2 and B2 matrices and in A2 and B10 matrices are investigated by the dissipative particle dynamics type of molecular-dynamics simulations. The monomer concentrations of A2B8(phiA2B8) studied are below the critical micelle concentration (phiA2B8(cmc)) for the formation of micelles in the presence of an adsorbed amphiphilic monolayer at the interface. Macrophase separation from the mixed phase to the segregated state with A-rich and B-rich coexisting phases and the segregation of A2B8 at the interface are thermodynamically gradual but are accompanied by a pronounced stretching and orientation of the constituent chains. The segregation of A2B8 at the interface broadens the interfacial region and reduces the interfacial tension. The chain conformation of the asymmetric amphiphilic molecules and the interfacial properties are dominated by the majority block in the amphiphilic chain and dependent on the composition of the matrix in contact with the majority block. In the A2 and B2 matrices, the B8 blocks in A2B8 chains at the interface resemble a wet brush swollen by short B2 chains. Swelling is responsible for the pronounced stretching and orienting of the amphiphilic chains and the reduced interfacial amphiphile enrichment. At the same interfacial amphiphile excess, however, swollen amphiphiles are more efficient in reducing the interfacial tension than nonswollen amphiphiles. PMID- 16375566 TI - Role of polydispersity in anomalous interactions in electrostatically levitated colloidal systems. AB - In this paper, we investigate the effects of using inverse analyses developed for monodisperse particles to extract particle-particle and particle-surface potentials from simulated interfacial colloidal configurations having finite-size polydispersity. Forward Monte Carlo simulations are used to generate three dimensional equilibrium configurations of log normal-distributed polydisperse particles confined by gravity near an underlying surface. Particles remain levitated above the substrate and stabilized against aggregation by repulsive electrostatic Derjaguin-Landau-Verwey-Overbeek pair potentials. An inverse Ornstein-Zernike analysis and an inverse Monte Carlo simulation method are used to obtain interactions from simulated distribution functions as a function of polydispersity (sigma), relative range of repulsion (kappa a), and projected interfacial concentration (rho). Both inverse analyses successfully recover input potentials for all monodisperse cases, but fail for polydispersities often encountered in experiments. For different conditions (sigma, kappa a, and rho), our results indicate softened short-range repulsion, anomalous long-range attraction, and apparent particle overlaps, which are similar to commonly reported observations in optical microscopy measurements of quasi-two-dimensional interfacial colloidal ensembles. By demonstrating signatures of, and limitations due to, polydispersity when extracting pair potentials from measured distribution functions, our specific goal is to provide a basis to objectively interpret and resolve the effects of polydispersity in optical microscopy experiments. PMID- 16375567 TI - Efficient Monte Carlo trial moves for polypeptide simulations. AB - A new move set for the Monte Carlo simulations of polypeptide chains is introduced. It consists of a rigid rotation along the (C(alpha)) ends of an arbitrary long segment of the backbone in such a way that the atoms outside this segment remain fixed. This fixed end move, or FEM, alters only the backbone dihedral angles phi and psi and the C(alpha) bond angles of the segment ends. Rotations are restricted to those who keep the alpha bond angles within their maximum natural range of approximately +/-10 degrees. The equations for the angular intervals (tau) of the allowed rigid rotations and the equations required for satisfying the detailed balance condition are presented in detail. One appealing property of the FEM is that the required number of calculations is minimal, as it is evident from the simplicity of the equations. In addition, the moving backbone atoms undergo considerable but limited displacements of up to 3 A. These properties, combined with the small number of backbone angles changed, lead to high acceptance rates for the new conformations and make the algorithm very efficient for sampling the conformational space. The FEMs, combined with pivot moves, are used in a test to fold a group of coarse-grained proteins with lengths of up to 200 residues. PMID- 16375568 TI - Signatures of the fast dynamics in glassy polystyrene: first evidence by high field electron paramagnetic resonance of molecular guests. AB - The reorientation of one small paramagnetic molecule (spin probe) in glassy polystyrene (PS) is studied by high-field electron paramagnetic resonance spectroscopy at two different Larmor frequencies (190 and 285 GHz). Two different regimes separated by a crossover region are evidenced. Below 180 K the rotational times are nearly temperature independent with no apparent distribution. In the temperature range of 180-220 K a large increase of the rotational mobility is observed with the widening of the distribution of correlation times which exhibits two components: (i) a deltalike, temperature-independent component representing the fraction of spin probes w which persist in the low-temperature dynamics; (ii) a strongly temperature-dependent component, to be described by a power distribution, representing the fraction of spin probes 1-w undergoing activated motion over an exponential distribution of barrier heights g(E). Above 180 K a steep decrease of w is evidenced. The shape and the width of g(E) do not differ from the reported ones for PS within the errors. For the first time the large increase of the rotational mobility of the spin probe at 180 K is ascribed to the onset of the fast dynamics detected by neutron scattering at T(f)=175+/-25 K. PMID- 16375569 TI - Dipolar interactions, molecular flexibility, and flexoelectricity in bent-core liquid crystals. AB - The effects of dipolar interactions and molecular flexibility on the structure and phase behavior of bent-core molecular fluids are studied using Monte Carlo computer simulations. Some calculations of flexoelectric coefficients are also reported. The rigid cores of the model molecules consist of either five or seven soft spheres arranged in a "V" shape with external bend angle gamma. With purely repulsive sphere-sphere interactions and gamma = 0 degrees (linear molecules) the seven-sphere model exhibits isotropic, uniaxial nematic, and untilted and tilted smectic phases. With gamma > or = 20 degrees the untilted smectic phases disappear, while the system with gamma > or = 40 degrees shows a direct tilted smectic-isotropic fluid transition. The addition of electrostatic interactions between transverse dipole moments on the apical spheres is generally seen to reduce the degree of molecular inclination in tilted phases, and destabilizes the nematic and untilted smectic phases of linear molecules. The effects of adding three-segment flexible tails to the ends of five-sphere bent-core molecules are examined using configurational-bias Monte Carlo simulations. Only isotropic and smectic phases are observed. On the one hand, molecular flexibility gives rise to pronounced fluctuations in the smectic-layer structure, bringing the simulated system in better correspondence with real materials; on the other hand, the smectic phase shows almost no tilt. Lastly, the flexoelectric coefficients of various nematic phases--with and without attractive sphere-sphere interactions- are presented. The results are encouraging, but a large computational effort is required to evaluate the appropriate fluctuation relations reliably. PMID- 16375570 TI - Three-dimensional dynamic Monte Carlo simulations of elastic actin-like ratchets. AB - We present three-dimensional dynamic Monte Carlo simulations of the growth of a semiflexible fiber against a fluctuating obstacle. The natural reference for our numerical study are the elastic and Brownian ratchet models previously analyzed semianalytically. We find that the decay of the velocity versus applied load is exponential to a good degree of accuracy, provided we include in the load the drag force felt by the moving obstacle. If the fiber and obstacle only interact via excluded volume, there are small corrections to the Brownian ratchet predictions which suggest that tip fluctuations play a minor role. If on the other hand fiber and obstacle interact via a soft potential, the corrections are much larger when the obstacle diffuses slowly. This means that microscopic assumptions can profoundly affect the dynamics. We also identify and characterize a novel "pushing catastrophe"--which is distinct from the usual fiber buckling- in which the growth of the fiber decouples from the obstacle movement. The time distribution of catastrophes can be explained via an approximate analytical treatment, and our numerics suggest that the time taken to lose propulsive force is largely dependent on the fiber incidence angle. Our results are a first step in realizing numerical polymer models for the motion of sets or networks of semiflexible fibers close to a fluctuating membrane or obstacle. PMID- 16375571 TI - Molecular dynamics simulation of discontinuous volume phase transitions in highly charged crosslinked polyelectrolyte networks with explicit counterions in good solvent. AB - The volumetric properties of highly-charged defect-free polyelectrolyte networks with tetrafunctional crosslinks are studied through molecular dynamics simulations in the canonical ensemble. The network backbone monomers, which are monovalent, and the counterions, which are mono-, di-, or trivalent, are modeled explicitly in the simulations, but the solvent is treated implicitly as a dielectric medium of good solvation quality. The osmotic pressure of the network solvent system is found to depend greatly on the strength of electrostatic interactions. Discontinuous volume phase transitions are observed when the electrostatic interactions are strong, and the onset of these transitions shifts to higher solvent dielectricity as the counterion valency increases. The roles of the various virial contributions to the osmotic pressure are examined. The network elasticity entropy is found to behave nearly classically. As the network contracts and collapses with increasing strength of electrostatic interactions, the loss of counterion entropy leads to increased counterion osmotic pressure contributions via two mechanisms. The reduction in available configurational space increases the counterion translational entropy contribution to the ideal part of the osmotic pressure, and the greater number of counterion-monomer contacts formed due to counterion condensation and confinement increases the counterion excluded-volume entropy contribution to the excess part of the osmotic pressure. These observations contrast the decrease in the single ideal-gas-like counterion translational entropy contribution to the osmotic pressure predicted by the counterion condensation-charge renormalization theory. An accompanying decrease in the total electrostatic energy balances the loss of counterion excluded-volume entropy as the polyelectrolyte networks collapse in low dielectric solvents. This interplay between the electrostatic energy and the counterion excluded-volume entropy appears to be responsible for the discontinuous volume phase transitions that are observed in polyelectrolyte networks. The structure of the polyelectrolyte network is also found to be affine in the swollen state, with constituent chains nearly fully extended, and nonaffine in the collapsed state, with the chains adopting a Gaussian conformation. PMID- 16375572 TI - A physical model for the longitudinal polarizabilities of polymer chains. AB - The aim of this work is to provide a physical model to relate the polarizability per unit cell of oligomers to that of their corresponding infinite polymer chains. For this we propose an extrapolation method for the polarizability per unit cell of oligomers by fitting them to a physical model describing the dielectric properties of polymer chains. This physical model is based on the concept of a dielectric needle in which we assume a polymer chain to be well described by a cylindrically shaped nonconducting rod with a radius much smaller than its length. With this model we study in which way the polarizability per unit cell approaches the limit of the infinite chain. We show that within this model the macroscopic contribution of the induced electric field to the macroscopic electric field vanishes in the limit of an infinite polymer chain, i.e., there is no macroscopic screening. The macroscopic electric field becomes equal to the external electric field in this limit. We show that this identification leads to a relation between the polarizability per unit cell and the electric susceptibility of the infinite polymer chain. We test our dielectric needle model on the polarizability per unit cell of oligomers of the hydrogen chain and polyacetylene obtained earlier using time-dependent current-density functional theory in the adiabatic local-density approximation and with the Vignale-Kohn functional. We also perform calculations using the same theory on truly infinite polymer chains by employing periodic boundary conditions. We show that by extrapolating the oligomer results according to our dielectric needle model we get good agreement with our results from calculations on the corresponding infinite polymer chains. PMID- 16375575 TI - A conceptual model for recruitment and retention: allied health workforce enhancement in Western Victoria, Australia. AB - Attracting and retaining allied health professionals in rural areas is a recognised problem in both Australia and overseas. Predicted increases in health needs will require strategic actions to enhance the rural workforce and its ability to deliver the required services. A range of factors in different domains has been associated with recruitment and retention in the allied health workforce. For example, factors can be related to the nature of the work, the personal needs, or the way an organisation is led. Some factors cannot be changed (eg geographical location of extended family) whereas others can be influenced (eg education, support, management styles). Recruitment and retention of allied health professionals is a challenging problem that deserves attention in all domains and preparedness to actively change established work practices, both individually as well as collectively, in order to cater for current and predicted health needs. Changes to enhance workforce outcomes can be implemented and evaluated using a cyclic model. The Allied Health Workforce Enhancement Project of the Greater Green Triangle University Department of Rural Health (GGT UDRH) is working towards increasing the number of allied health professionals in the south west of Victoria. Based on themes identified in the literature, an interactive model is being developed that addresses recruitment and retention factors in three domains: (1) personal or individual; (2) organisation; and (3) community. PMID- 16375576 TI - Where does the evidence come from? PMID- 16375577 TI - NASBA: a novel approach to assess hormonal receptors and ERBB2 status in breast cancer. AB - In human breast cancer, estrogen receptor-alpha (ERalpha), progesterone receptor (PR) and human epidermal growth factor receptor (ERBB2) status are currently determined using different techniques. We propose to assess the mRNA expression of these three clinically relevant markers using a unique technique, real-time nucleic acid sequence-based amplification (NASBA). Gene expression of hormone receptors was analyzed and compared to the cytosolic functional protein content as determined with a ligand binding assay (LBA), while ERBB2 mRNA expression was compared to quantitative PCR and ELISA. We observed that the three markers are significantly overexpressed at the mRNA level in positive tumors, as measured by DNA- or protein-based techniques. Biostatistical analysis of the receiver operating characteristic (ROC) curve demonstrated high concordance between NASBA and LBA [area under the curve (AUC) for ROC of 0.899] and showed that ERalpha status could be predicted using the molecular assay with a sensitivity of 72.7% and a specificity of 93.5%. Similar results were obtained for PR (AUC ROC 0.938, sensitivity 75.3%, specificity 100%). Moreover, excellent concordance was observed between NASBA, quantitative PCR and ELISA with respect to ERBB2 (AUC ROC 0.92, sensitivity 90%, specificity 89.7%; and AUC ROC 0.98, sensitivity 100%, specificity 91.5%, respectively). These results suggest that NASBA is well suited for assessing ER, PR and ERBB2 status in breast tumor samples. This approach is rapid, highly sensitive and a standardized method that could be complementary to the existing techniques, especially for small tumors. PMID- 16375578 TI - Plasma cell-free DNA as an indicator of severity of injury in burn patients. AB - BACKGROUND: Raised levels of plasma cell-free DNA have been detected in various patient groups, including trauma patients. We hypothesized that plasma DNA is increased in burn patients and may represent an objective indicator of burn severity and have predictive as well as prognostic significance. METHODS: This was a prospective clinical study with full ethical approval. With informed consent, blood samples were collected from 28 burn patients within 24 h of injury and from 12 control subjects. Plasma cell-free DNA was measured by real-time quantitative polymerase chain reaction (PCR) assay for the beta-globin gene. Descriptive analysis, non-parametric data comparison tests (Mann-Whitney) and correlation tests (Spearman rank) were performed on the data. RESULTS: Samples were taken at a mean time of 5.7 h after injury from 13 patients with flame/flash burns and 15 patients with scalds. Median plasma DNA levels in the control, scald and flame/flash burn patient groups were 287, 648 and 2685 kilogenome equivalents/L, respectively. Plasma DNA levels correlated with the length of hospital stay, but not with admission to the intensive care unit (ICU) nor the length of ICU stay. DNA levels correlated with the burn surface area (Spearman rank r = 0.54, p = 0.04) and the number of operations needed (Spearman rank r = 0.55, p = 0.03) for scalds, but not for flame/flash burns. CONCLUSIONS: Plasma DNA is increased after burn injury and is significantly correlated with some outcome measures, including the length of hospital stay. DNA levels are higher in flame/flash patients than in scald patients; the difference may provide an objective indication of burn depth and inhalation injury. PMID- 16375579 TI - Bivariate statistical approach to evaluate laboratory performance by analysis of standard curves in an External Quality Assurance program for quantitative assays based on real-time PCR with Taq-Man probes. AB - Recently a revolutionary technique for quantitative PCR determination was introduced in diagnostic laboratories. To determine the influence of technical variability on the reliability of the quantitative assay, it is crucial to use External Quality Assurance (EQA) programs. An EQA program was developed in Italy to check the analytical performance of real-time PCR procedures based on Taq Mantrade mark probes. This article suggests a new statistical approach to discriminate, using a bivariate technique, laboratory performance that appears to be questionable, by separately considering the two main features of the standard curve: analytical sensitivity and efficiency. Furthermore, specific indexes to evaluate the impact of these two features on the determination of the initial number of molecules are given to help to improve the assay procedure. PMID- 16375580 TI - Erythrocyte membrane acetylcholinesterase activity in subjects with MTHFR 677C- >T genotype. AB - BACKGROUND: Increased homocysteine (Hcy) blood levels are correlated with vascular and neurological problems. AIM: The aim of the present study was to investigate erythrocyte membrane acetylcholinesterase (AChE) activity in subjects with the MTHFR C677T genotype in relation to Hcy. METHODS: Blood was obtained from 22 individuals with the MTHFR C677T genotype before and after folic acid supplementation and once from controls (n = 30). Plasma folate, vitamin B(12) and total antioxidant status (TAS) were measured with commercial kits, Hcy by a HPLC method and membrane enzyme activity spectrophotometrically. RESULTS: In MTHFR C677T carriers, AChE activity was significantly higher (4.20 +/- 0.12 x mg protein) and decreased to normal levels (3.14 +/- 0.10 x mg protein; p < 0.001) after therapy. TAS differed slightly before and after treatment. Hcy levels were significantly higher before (22.4 +/- 2.8 microM) compared to after (12.1 +/- 2.0 microM; p < 0.001) therapy and compared to controls (10.5 +/- 2.5 micromol/L; p < 0.001). In an in vitro study, incubation of Hcy-activated membrane AChE from controls with phenylalanine resulted in restoration of activity, but failed to reverse the stimulated enzyme from hyperhomocysteinaemic MTHFR C677T subjects before therapy. Alanine incubation protected the enzyme from Hcy activation in controls. CONCLUSIONS: Increased membrane AChE activity may be due to high Hcy levels. In vitro, phenylalanine reversed the Hcy activation of the membrane enzyme from controls and alanine protected it from Hcy action. PMID- 16375581 TI - Plasma levels of tumor M2-pyruvate kinase should not be used as a tumor marker for hematological malignancies and solid tumors. AB - It has been reported that the dimeric isoform of the enzyme pyruvate kinase M2 was overexpressed in various solid tumor cells. Hence, it was suggested that circulating levels of the so-called tumor M2-pyruvate kinase (Tu M2-PK) could be used as a tumor marker for monitoring systemic therapies of various solid tumors. We analyzed its validity as a tumor marker by comparing plasma levels of Tu M2-PK in patients with different non-malignant diseases to levels in healthy individuals and in patients with hematological diseases. Plasma levels of Tu M2 PK were measured using an ELISA assay in a total of 284 patients. The mean Tu M2 PK concentration of 32 U/mL was significantly higher in the group of patients with hematological malignancies (n = 121) (p < 0.001). However, 37% of healthy individuals (n = 63) and 44% of patients with non-malignant diseases (n = 100), especially patients with an acute inflammatory reaction (67%), were found to have elevated levels of Tu M2-PK using a cutoff level of 15 U/mL. The specificity was 59% and the sensitivity was 51%. There was no significant correlation between the prevalence of a hematological malignancy and positive Tu M2-PK result. Thus, our data imply that Tu M2-PK is not a useful tumor marker for hematological malignancies and solid tumors, as a significant number of false positive results were detected in healthy individuals and patients with non-malignant diseases. PMID- 16375583 TI - Association study between fibronectin and coronary heart disease. AB - Fibronectin is a known chemoattractant for several cell types that play a role in the wound healing process, including fibroblasts, endothelial cells and macrophages. It also generates a scaffold that allows attachment of other extracellular matrix components. Large amounts of fibronectin have been detected in atherosclerotic plaques, suggesting that it may play a role in the pathogenesis of atherosclerosis. To examine the possible involvement of fibronectin in the etiology of atherosclerotic coronary heart disease, we analyzed four polymorphisms in the human fibronectin gene and determined the plasma fibronectin levels in patients with coronary heart disease (n = 109) and age- and gender-matched controls (n = 123) in Chinese Han people. No significant positive association was observed between these polymorphisms and coronary heart disease. The levels of circulating plasma fibronectin, however, were significantly lower in patients with coronary heart disease (mean +/- SD 245 +/- 87 mg/L) compared with controls (354 +/- 88 mg/L) (p < 0.001). The odds ratio (OR) for plasma fibronectin was 0.94 in a multivariate unconditional logistic regression model (OR = 0.94, 95% CI 0.91-0.96, p < 0.001). We conclude that, in our population, the four fibronectin gene polymorphisms detected are not associated with clinical coronary heart disease. Our data suggest that low circulating fibronectin levels might be a new marker of coronary heart disease. PMID- 16375582 TI - APO A-V-1131T-->C polymorphism frequency and its association with morbidity in a Brazilian elderly population. AB - Identification of genetic polymorphisms as risk factors for complex diseases affecting older people can be relevant for their prevention, diagnosis and management. The -1131T-->C polymorphism of the apolipoprotein A-V gene (APO A-V) is tightly linked to lipid metabolism and has been associated with increased triglyceride levels and familial dyslipidemia. The aims of this study were to analyze the allele and genotype frequencies of this polymorphism in a Brazilian elderly population and to investigate any association between the polymorphism and major morbidities affecting elderly people. This polymorphism was investigated in 371 individuals, aged 66-97 years, in a Brazilian Elderly Longitudinal Population Study. Major morbidities investigated were: cerebrovascular diseases (CVD); myocardial infarction (MI); type 2 diabetes; hypertension; obesity; dementia; depression; and neoplasia. DNA was isolated and amplified by PCR and its products were digested with restriction enzyme Tru1I. T and C allele frequencies were 0.842 and 0.158, respectively. Our population showed allele frequencies that were similar to European and Afro-American and different from Asiatic populations. Genotype distributions were not within Hardy Weinberg equilibrium only for the obesity subject sample. On the other hand, a significant association between the C allele and obesity in the presence of CVDxdepression interaction was observed. Logistic analysis showed no association of the polymorphism with each morbidity group. Therefore, the C allele in elderly Brazilian subjects may represent a risk factor for these morbidity interactions, which may lead to better comprehension of their pathophysiology. PMID- 16375584 TI - Serum calcium and phosphorus associate with the occurrence and severity of angiographically documented coronary heart disease, possibly through correlation with atherogenic (apo)lipoproteins. AB - The associations of serum calcium and phosphorus concentrations as well as other cardiovascular risk factors were investigated in relation to the existence and severity of coronary heart disease (CHD) in 260 clinically stable, angiographically defined CHD patients aged 40-70 years. The subjects were classified as CHD(+) cases if one or more coronary arteries had a significant stenosis (> or =70%) and CHD(-) controls if there was no stenosis (< or =10%) in any artery. The severity of coronary occlusion was scored on the basis of the number and extent of lesions, as normal, mild, moderate or severe. Fasting serum concentrations of electrolytes, lipids and (apo)lipoproteins were determined. The concentrations of serum total calcium (2.41 +/-0.14 vs. 2.33 +/- 0.22 mmol/L, p < or = 0.05), albumin-corrected calcium (2.33 +/- 0.25 vs. 2.23 +/- 0.25 mmol/L, p < or = 0.01), phosphorus (1.32 +/-0.21 vs. 1.25 +/- 0.17 mmol/L, p < or = 0.007) and the ion product of calcium and phosphorus (3.16 +/- 0.58 vs. 2.91 +/- 0.50, p < or =0.0001) were significantly higher in the CHD(+) compared to the CHD(-) group. Patients with CHD compared with controls had increased serum levels of triglyceride, total cholesterol, low-density lipoprotein-cholesterol (LDL-C), apolipoprotein B (apoB), lipoprotein(a) [Lp(a)] and decreased serum levels of high-density lipoprotein (HDL)-C and apoAI. Multiple logistic regression analysis showed strong and significant association between diabetes mellitus (odds ratio, OR = 5.24, p < or = 0.0001), male gender (OR = 8.84, p < or =0.0001), Lp(a) (OR = 1.014, p < or =0.006), hypertension (OR = 2.61, p < or =0.02), apoB (OR = 1.031, p < or =0.001), age (OR = 1.055, p < or =0.003), phosphorus (OR = 2.438, p < or =0.01), albumin-adjusted calcium (OR = 1.532, p < or =0.05), cholesterol (OR = 1.009, p < or =0.05) and the occurrence of CHD. On the basis of bivariate correlation analysis, serum-adjusted calcium was positively correlated with the levels of cholesterol (r = 0.285, p < or =0.0001), LDL-C (r = 0.320, p < or =0.0001), Lp(a) (r = 0.173, p < or = 0.005), apoB (r = 0.237, p < or =0.0001), LDL-C/apoB ratio (r = 0.180, p < or= 0.007), apoAI (r = 0.181, p < or =0.003) and inversely to HDL-C (r = -0.146, p < or =0.02) and HDL-C/apoAI ratio (r = -0.263, p < or =0.0001). Serum phosphorus concentration was a significant correlate of triglyceride (r = 0.199, p < or =0.001) and Lp(a) (r = 0.129, p < or =0.04). The results demonstrated that serum calcium and phosphorus are associated with the prevalence and severity of CHD, probably through correlation with atherogenic lipids and (apo)lipoproteins. Serum calcium and phosphorus and their ion product were also independent risk factors for CHD. PMID- 16375585 TI - Urinary calcium excretion in severe preeclampsia and eclampsia. AB - Urinary calcium levels in women with mild preeclampsia, severe preeclampsia and eclampsia were evaluated in this study. We collected 24-h urine samples from 35 mild preeclamptic (Group 1), 30 severe preeclamptic (Group 2), and 17 eclamptic patients (Group 3). The control group (Group 4) consisted of 35 healthy pregnant women. Serum levels of total calcium and creatinine, and urinary calcium were measured. These values were compared in the four groups. The mean maternal age and parity were similar in all groups. There were no statistically significant differences in the serum levels of total calcium and creatinine (p > 0.05). Urinary calcium excretion in patients with preeclampsia and eclampsia was significantly lower than in controls (p < 0.0001). Urinary calcium levels between mild preeclampsia and severe pre-eclampsia, and severe preeclampsia and eclampsia were similar (p > 0.05), but were lower in eclampsia than in mild preeclampsia (p < 0.05). In conclusion, urinary calcium excretion is reduced in patients with severe preeclampsia or eclampsia. However, the decrease in urinary calcium excretion cannot be used to identify the severity of preeclampsia, or to predict impending eclampsia. PMID- 16375586 TI - In vitro re-mineralization of demineralized bone matrix in human serum. AB - The aim of this study was to develop an in vitro re-mineralization model in human serum. For this purpose a commercially produced demineralized human bone matrix (DBM) was incubated in samples of human serum pools obtained from two physiologically different groups. The first group consisted of young males and the second of older females. After incubation periods of 4 and 7 days at 37 degrees C, changes in the levels of calcium and inorganic phosphate content of the serum and DBM samples were measured. The results of the study showed that the change in mineral content of serum and DBM samples in both study groups was statistically significant. The decrease in serum calcium content and increase in DBM inorganic phosphate content were significant in the young group for longer incubation times. In the older group, both serum calcium and inorganic phosphate decreased and DBM mineral content increased for the same incubation time. When the two physiological groups were compared, statistically significant differences were identified for changes in mineral levels in both serum and the DBM samples. These data indicate that the mineral content of human serum decreases and that of DBM increases when these two materials are incubated together. These changes provide evidence for the re-mineralization of DBM. The model described here could also detect a difference in re-mineralization capability between two different groups of human sera. PMID- 16375587 TI - Measurement of serum amyloid A1 (SAA1), a major isotype of acute phase SAA. AB - Serum amyloid A (SAA), a plasma precursor of reactive amyloid deposits, is a multigene product. SAA1 and SAA2, with primary structures that are 93% identical (98 of 104 amino acids), behave as acute phase proteins, as demonstrated by their increasing levels in plasma. Heretofore, it has been understood that SAA1 predominates and functions as an isotype in plasma. However, accurate measurements differentiating the two isotypes have not been reported. In this study, using monoclonal antibodies specific for SAA1, we developed an enzyme linked immunosorbent assay (ELISA) for SAA1. The levels and ratios of SAA1 in total SAA (TSAA) were investigated in healthy subjects and patients with rheumatoid arthritis (RA). The SAA1/TSAA ratio was 74 +/- 12% and 77 +/- 12% in healthy subjects and RA patients, respectively. In RA patients, the ratios were not influenced by SAA1 genotype, which has been proposed to affect plasma SAA values. The kinetics of SAA1 in inflamed patients undergoing hemodialysis was found to be parallel with total SAA and C-reactive protein. Finally, this study confirmed that SAA1 is a major isotype of acute phase SAA and may determine total SAA values. This specific assay could be used in the evaluation of SAA behavior in several clinical conditions. PMID- 16375588 TI - Disturbed lipoprotein composition in non-dialyzed, hemodialysis, continuous ambulatory peritoneal dialysis and post-transplant patients with chronic renal failure. AB - Studies were carried out in 183 non-dialyzed, 123 hemodialysis, 81 continuous ambulatory peritoneal dialysis and 35 post-transplant patients and in 103 healthy subjects as a reference group. Lipids and apolipoprotein (apo)AI and apoB were determined using Roche kits. An anti-apoB antibody was used to separate apoB containing apoCIII and apoE-triglyceride-rich lipoprotein (TRL) in the non-high density lipoprotein (non-HDL) fraction from apoCIIInonB and apoEnonB in the HDL fraction in four groups of patients with chronic renal failure (CRF) and healthy subjects. Multivariate linear regression analysis was used to investigate the relationship between triglyceride (TG) or HDL-cholesterol (HDL-C) concentrations and lipoproteins. Dyslipidemia varied according to the degree of renal insufficiency, the type of dialysis and therapy regime in CRF patients. Lipoprotein disturbances were manifested by increased TG, non-HDL-C and TRL concentrations, and decreased HDL-C and apoAI concentrations, whereas post-renal transplant patients showed normalization of lipid and lipoprotein profiles, except for TG levels and total apoCIII and apoCIIInonB. The present study indicates that CRF patients have disturbed lipoprotein composition, and that hypertriglyceridemia and low HDL-C concentrations in these patients are multifactorial, being secondary to disturbed lipoproteins. The method using anti apoB antibodies to separate apoB-containing lipoproteins in the non-HDL fraction from non-apoB-containing lipoproteins in HDL can be used in the diagnosis and treatment of patients with progression of renal failure or atherosclerosis. The variability of TG and HDL-C concentrations depends on the variability of TRL and cholesterol-rich lipoprotein concentrations, but the decreases in TG and increases in HDL-C concentrations are caused by apoAI concentration variability. These relationships, however, need to be confirmed in further studies. PMID- 16375589 TI - Measurement of serum testosterone using high-performance liquid chromatography/tandem mass spectrometry. AB - Low levels of serum testosterone typically found in women and children cannot be reliably measured by immunoassay. We developed a simple and sensitive method using high-performance liquid chromatography/tandem mass spectrometry (HPLC MS/MS). Sample preparation involved protein precipitation of serum (1.0 mL) with acetonitrile containing the internal standard (testosterone-d3) followed by liquid extraction with methylene chloride. The chromatographic cycle per specimen was 10 min. The performance was evaluated according to the CLSI EP10-A2 protocol. Within- and between-run imprecision was 20.9%, 2.29% and 1.80%, and 1.81%, 3.58% and 2.97% at mean concentrations of 0.17, 14.1 and 28.8 nM, respectively, with no apparent carryover. The method was linear from 0.21 to 53.1 nM and the analytical recovery was 100.5-106.2% across the concentrations tested. There was no interference observed from other steroids that were tested. Correlation using de identified patient specimens with a commercial HPLC-MS/MS method showed a slope of 0.991, an intercept of -0.017 and a correlation coefficient (R(2)) of 0.998 by linear regression over concentrations ranging from 0.21 to 16.7 nM. In conclusion, we report here an HPLC-MS/MS method suitable for clinical measurement of serum testosterone. PMID- 16375590 TI - Quantitative bacterial micro-assay for rapid detection of serum phenylalanine on dry blood-spots: application in phenylketonuria screening. AB - Phenylketonuria is an inherited metabolic disease, which is characterized by an increased level of serum phenylalanine. Quantitative measurement of phenylalanine in the serum of phenylketonuria patients is necessary to confirm the disease, and to distinguish phenylketonuria from other forms of hyperphenylalaninemia. In this study, we report a rapid and inexpensive micro-assay for simultaneous detection and quantitative measurement of serum phenylalanine on dry blood-spots. Analysis of the standard curve showed a broad linear range for phenylalanine from 120 to 1800 micromol/L. Application of this method, the standard Guthrie bacterial inhibition assay and a high-performance liquid chromatography (HPLC) method for analysis of 34 samples from phenylketonuria patients and control samples produced comparable results, with the regression equation Y = 0.994X + 0.996. The advantage of this method over the Guthrie bacterial inhibition assay is its ability to measure serum phenylalanine quantitatively without false positive results. The method was successfully applied to dried blood-spots, serum and whole blood. The cost per sample is approximately 20-50 US cents, which is much less than for HPLC and commercial enzyme kits. The method can be automated, and is thus suitable for neonatal and mass screening for phenylketonuria, especially in developing countries where funding is a limiting factor. PMID- 16375591 TI - N-Terminal pro-brain natriuretic peptide: normal ranges in the pediatric population including method comparison and interlaboratory variability. AB - The aim of the present study was the investigation of N-terminal pro-brain natriuretic peptide (NT-proBNP) in the pediatric population. This is essential for adequate monitoring and classification of pediatric patients with heart disease, but no consistent data are available yet. In addition, the comparability of two commercially available NT-proBNP assays and the inter-laboratory variability for the most suitable one were assessed. For this purpose, 408 subjects (1-29 years) were included. NT-proBNP was determined with a non competitive electrochemiluminescent immunoassay (Roche NT-proBNP; n = 402) and a competitive enzyme-immunoassay (Biomedica NT-proBNP; n = 402). Inter-laboratory variability was evaluated for the Roche assay by stepwise inclusion of four and 11 centers throughout Germany, respectively. Roche NT-proBNP ranged from 5.0 to 391.5 ng/L, with higher values for younger children. The 97.5th (75th) percentile curve ranged from 319.9 ng/L (231.2 ng/L, 1-3 years) to 114.9 ng/L (53.3 ng/L, 18 years). In contrast, Biomedica NT-proBNP ranged from 253.7 to 7602.8 ng/L, with no significant age dependency. The mean difference between the assays was 1649.7 ng/L (95% confidence interval 1546.3-1753.1 ng/L). Inter-laboratory variability ranged from 6.5% to 3.8%, covering a range from 51.3 to 6618.1 ng/L. The assay seems to influence the interpretation of resulting NT-proBNP values and therefore has to be chosen carefully. For the monitoring and classification of pediatric patients with congenital heart disease, age-based NT-proBNP values should be used. PMID- 16375592 TI - A new quality control model using performance goals based on biological variation in External Quality Assurance Schemes. AB - A new quality control model using performance goals based on biological variation in External Quality Assurance Schemes (EQAS) is described. The proposed model aims to use assay (analytical) CV(A), bias and total error available from participation in EQAS to describe assay performance using minimum, desirable and optimum quality specifications based on biological variation. The model provides further analysis of EQAS data and should be useful in better management of laboratory quality control, as it provides further information that can facilitate trouble-shooting. Additionally, it can help in evaluating the performance of current and proposed new laboratory methods by applying a unifying system if different EQAS are used to cover a range of analytes. PMID- 16375593 TI - Improvement in glycemic control over 11 years in patients monitored for diabetes in one county. AB - BACKGROUND: Hemoglobin A(1c) (HbA(1c)) has been used in controlled trials for the last 10 years but has never been evaluated in clinical practice as an effective parameter for clinical outcome. We investigated the trend for glycemic control over 11 years in one county of 350,000 citizens. METHODS: We studied 226,382 HbA(1c-DCCT-aligned) from 39,455 patients in whom routine monitoring for diabetes was initiated in 1993, 1996, or 2001. The trend in glycemic control was investigated in groups by probit plots, and in individual patients by target plots. RESULTS: From 1993 to 2001, the number of HbA(1c) measurements increased three-fold. The number of new monitoring series increased from 0.22% to 0.27% of the county population, and the number of patients monitored using HbA(1c) as a proxy for diabetes increased from 0.5% to 1.5%. A proportional reduction in high HbA(1c) concentrations of 5% was identified in the 1993 group, compared to 15% in the 1996 group, and 20% in the 2001 group. The percentage of patients with diabetic first HbA(1c) experiencing normalization increased from 8% to 30% for males and from 9% to 24% for females (1993-2001). The percentage of HbA(1c) concentrations that were not normalized decreased from 78% to 53% for males and from 83% to 59% for females (1993-2001). The median HbA(1c) at initiation of monitoring decreased from 8.7% in 1993 to 7.5% in 2001 (p < 10(-5)). The number of normal first HbA(1c) results in monitoring series increased from 7% to 17% for males and from 8% to 22% for females. Up to 10% of subjects developed diabetic concentrations during monitoring. CONCLUSION: On average, patients with diabetic first HbA(1c) concentrations (> or =6.62%) showed an improvement in glycemic control from 5% in 1993 to 20% in 2001. High concentrations were easiest to reduce. In patients with originally diabetic HbA(1c) levels, 66% on average showed improved glycemic control in the 2001 series compared to 50% in the 1993 series. An average of 6% (1993) vs. 9% (2001) with originally normal HbA(1c) levels showed an upward trend inHbA(1c) levels. Median HbA(1c) at initiation of monitoring decreased from 8.7% in 1993 to 7.5% in 2001 (p < 10(-5)). The incidence of new cases was constant. PMID- 16375594 TI - The clinical usefulness of glucose tolerance testing in gestational diabetes to predict early postpartum diabetes mellitus. AB - We examined the clinical usefulness of antepartum clinical characteristics, along with measures of glucose tolerance, in Dutch multiethnic women with gestational diabetes mellitus (GDM) for their ability to predict type 2 diabetes within 6 months of delivery (early postpartum diabetes). The present study comprised a cross-sectional 5-year investigation (1998-2003) of a consecutive series of 168 women with GDM identified by a two-stage protocol at 16-33 weeks of gestation. The following data were collected for all women: age and gestational age at entry into the study; prepregnancy body mass index (BMI); ethnicity; obstetric and clinical history, including the onset of early postpartum diabetes; pregnancy outcome; level of fasting C-peptide; and glycemic parameters of 50-g 1-h glucose challenge test and 100-g 3-h oral glucose tolerance test (diagnostic OGTT). We used receiver operating characteristic (ROC) curve analysis to test the clinical usefulness of the glycemic parameters. A total of 11 women (6.5%) developed early postpartum diabetes. Apart from family history of diabetes (p = 0.052), anthropometric, maternal, and neonatal clinical parameters showed no association with early postpartum diabetes in univariate analyses. The level of fasting glucose, and both the glucose challenge test and diagnostic OGTT post-load glucose levels and glucose areas were associated with early postpartum diabetes. ROC curve analysis identifiedall three glucose challenge-test parameters, including fasting glucose concentration, as poor diagnostic tests, with a positive predictive value of approximately 22%, whereas the positive predictive value associated with the area under the diagnostic OGTT curve increased progressively over monitoring time from 20.6% to 100%. Using a 3-h OGTT glucose area threshold of 35.7 mmol.h/L resulted in 100% sensitivity and 100% specificity, identifying the 11 women who developed early postpartum diabetes. In summary, we can conclude from the present analysis that early postpartum diabetes is rare in GDM women (6.5%), and that the clinical usefulness of the total area under the diagnostic 3-h OGTT is superior to all other glycemic parameters for detecting early postpartum diabetes. PMID- 16375595 TI - Analytical performance of a new two-step ADVIA Centaur estradiol immunoassay during ovarian stimulation. AB - Measurement of estradiol is useful in women undergoing ovarian stimulation for in vitro fertilization and embryo transfer (IVF-ET). The analytical performance of a new two-step estradiol assay (ADVIA Centaur) estradiol-6 III from Bayer Diagnostics) was evaluated in 41 sera from 11 women undergoing ovarian stimulation. The results were compared to those obtained with two radioimmunoassays (RIAs; RIA Estradiol Immunotech IM 1663 from Beckmann Coulter and Coat-A-Count Estradiol from Diagnostic Products Corporation) and with one chemiluminescent immunoassay (CLIA; ADVIA Centaur estradiol-6). The ADVIA Centaur) estradiol-6 III assay was the most sensitive assay, with a functional sensitivity of 55 pmol/L. Within- and between-run coefficients of variation calculated for the new ADVIA Centaur assay ranged from 3.3% to 9%, which was better than the precision obtained for the other assays. A dilution test showed serum interferences when estradiol was measured in non-diluted samples. No statistical difference was observed between the estradiol results obtained in diluted sera with the new two-step ADVIA Centaur assay and those measured with the Immunotech RIA and the other CLIA. In conclusion, this new, two-step estradiol assay performed on the ADVIA Centaur system displays suitable sensitivity, precision and intermethod agreement with the Immunotech RIA for the measurement of serum estradiol concentrations in women undergoing ovarian stimulation and IVF-ET. For correct linearity, estradiol measurement should be performed on diluted samples. PMID- 16375596 TI - EC4 European Syllabus for Post-Graduate Training in Clinical Chemistry and Laboratory Medicine: version 3 - 2005. AB - The EC4 Syllabus for Postgraduate Training is the basis for the European Register of Specialists in Clinical Chemistry and Laboratory Medicine. The syllabus: Indicates the level of requirements in postgraduate training to harmonise the postgraduate education in the European Union (EU); Indicates the level of content of national training programmes to obtain adequate knowledge and experience; Is approved by all EU societies for clinical chemistry and laboratory medicine. The syllabus is not primarily meant to be a training guide, but on the basis of the overview given (common minimal programme), national societies should formulate programmes that indicate where knowledge and experience is needed. The main points of this programme are: Indicates the level of requirements in postgraduate training to harmonise the postgraduate education in the European Union (EU); Indicates the level of content of national training programmes to obtain adequate knowledge and experience; Is approved by all EU societies for clinical chemistry and laboratory medicine. Knowledge in biochemistry, haematology, immunology, etc.; Pre-analytical conditions; Evaluation of results; Interpretations (post analytical phase); Laboratory management; and Quality insurance management. The aim of this version of the syllabus is to be in accordance with the Directive of Professional Qualifications published on 30 September 2005. To prepare the common platforms planned in this directive, the disciplines are divided into four categories: Indicates the level of requirements in postgraduate training to harmonise the postgraduate education in the European Union (EU); Indicates the level of content of national training programmes to obtain adequate knowledge and experience; Is approved by all EU societies for clinical chemistry and laboratory medicine. Knowledge in biochemistry, haematology, immunology, etc.; Pre analytical conditions; Evaluation of results; Interpretations (post-analytical phase); Laboratory management; and Quality insurance management. General chemistry, encompassing biochemistry, endocrinology, chemical (humoral), immunology, toxicology, and therapeutic drug monitoring; Haematology, covering cells, transfusion serology, coagulation, and cellular immunology; Microbiology, involving bacteriology, virology, parasitology, and mycology; Genetics and IVF. PMID- 16375597 TI - POX-Act assay and d-ROMs test - what are the facts? PMID- 16375598 TI - External quality control of urinary methyl malonic acid quantification - announcement of a pilot study. PMID- 16375600 TI - Introduction: interferon signals: what is classical and what is nonclassical? PMID- 16375601 TI - Stats: multifaceted regulators of transcription. AB - The high-affinity binding interactions between interferons (IFNs) and their cognate cell surface receptors lead to the activation of receptor-associated Janus protein tyrosine kinases (Jaks) and subsequent phosphorylation and activation of a group of transcription factors, the signal transducers and activators of transcription (Stats). Upon IFN-induced activation, these Stat proteins form homodimeric and heterodimeric complexes that translocate to the nucleus and bind specific elements within the promoters of IFN-stimulated genes (ISGs). In addition to the well-studied IFN-induced ISG factor 3 (ISGF3) and Stat1:1 complexes, IFNs induce the formation of a number of other Stat-containing complexes, including Stat3:3 and Stat5:5 homodimers, as well as Stat2:1 and Stat5:CrkL heterodimers, that also mediate gene transcription. Moreover, emerging evidence suggests that particular amino acid residues within the individual Stat proteins contribute to different aspects of Stat function. These residues modulate the transcriptional activation potential of Stat-containing complexes and thereby influence the expression of ISGs. Indeed, the Stat proteins function in a multifaceted manner to regulate the expression of proteins that mediate IFN responses. PMID- 16375602 TI - Unexpected roles for deacetylation in interferon- and cytokine-induced transcription. AB - Protein acetylation is a reversible modification that has been implicated in epigenetic regulation of gene expression. It is widely accepted that acetylation enzymes are present at transcriptionally active promoters and deacetylation enzymes associate with transcriptionally silent loci. These results notwithstanding, recent findings indicate that positive regulation of gene expression by interferons and other cytokines requires both acetylation and deacetylation. PMID- 16375603 TI - The p38 mitogen-activated protein kinase pathway in interferon signal transduction. AB - Interferons (IFNs) are cytokines that regulate a variety of biologic effects, including cellular antiviral responses, inhibition of proliferation, induction of differentiation, and immunoregulation, via different mechanisms. In order to mediate such pleiotropic effects, IFNs trigger numerous signaling events. One way for IFNs to regulate cellular functions is through activation of mitogen activated protein (MAP) kinases. Three major cascades of MAP kinases are known. The c-Jun NH(2)-terminal kinase (JNK) cascade, the extracellular signal-regulated kinase (ERK) cascade, and the p38 MAP kinase cascade. ERK and p38 MAP kinases are activated in response to type I IFNs and participate in the regulation of cellular responses. In this review we discuss recent findings on the role of the p38 MAP kinase pathway and its function in mediating IFN-dependent biologic effects. We further dissect and discuss the roles of upstream and downstream components of the p38 MAP kinase in the control of cellular responses triggered by IFNs. PMID- 16375604 TI - CCAAT/enhancer binding proteins and interferon signaling pathways. AB - Interferons (IFNs) regulate a number of host responses, including innate and adaptive immunity against viruses, microbes, and neoplastic cells. These responses are dependent on the expression of IFN-stimulated genes (ISGs). Given the diversities in these responses and their kinetics, it is conceivable that a number of different factors are required for controlling them. Here, we describe one such pathway wherein transcription factor CAAAT/enhancer binding protein-beta (C/EBP-beta) is controlled via IFN-gamma-induced MAPK signaling pathways. At least two IFN-gamma-induced MAPK signals converge on to C/EBP-beta for inducing transcription. One of these, driven by extracellular signal-regulated kinases (ERKs), phosphorylates the C/EBP-beta protein in its regulatory domain. The second, driven by the mixed-lineage kinases (MLKs), induces a dephosphorylation leading to the recruitment of transcriptional coactivators. PMID- 16375605 TI - Immune cell-specific amplification of interferon signaling by the IRF-4/8-PU.1 complex. AB - Both type I interferon (IFN-alpha/beta) and type II IFN (IFN-gamma) exert many functions that are restricted to immune cells. Thus, they play critical roles in innate and adaptive immunity. IFN regulatory factor-4 (IRF-4) and IRF-8 (formerly PU.1 interaction partner [Pip] and IFN consensus sequence binding domain [ICSBP], respectively) are immune cell-specific members of the IRF family that regulate the development of myeloid, lymphoid, and dendritic cells. They form a heterodimeric complex with another immune cell-specific transcription factor PU.1 Spi-1 and regulate transcription of genes in the immune system. This review describes the role of the IRF-8-PU.1 complex in modulating IFN signaling in an immune cell-specific manner. Our studies revealed that some but not all IFN-gamma inducible genes carry an IFN-gamma activation site (GAS) element that contains a binding site for the IRF- 8-PU.1 complex. The IRF-8-PU.1 complex can take part in GAS-mediated transcription and amplify expression of IFN-gamma-responsive genes initiated by Stat1 in macrophages. Similarly, some but not all IFN-alpha/beta responsive genes are shown to carry an IFN-stimulated response element (ISRE) that contains an IRF-8-PU.1 binding site. The participation of IRF-8-PU.1 in ISRE mediated transcription results in the augmentation of IFN-stimulated gene factor 3 (ISGF3)-induced transcription in macrophages. Thus, GAS and ISRE elements, classically defined as universal IFN-alpha/beta and IFN-gamma response sequences, are not the same, and some harbor an embedded motif for IRF- 8-PU.1 binding that functions only in immune cells. Accordingly, the IRF-8-PU.1complex provides secondary IFN signaling pathways unique to the immune system. Collectively, the contribution of IRF-8 and PU.1 to IFN-regulated gene expression may in part account for immune cell-specific functions of IFNs. PMID- 16375606 TI - The PI3' kinase pathway in interferon signaling. AB - Interferons (IFNs) are pleiotropic cytokines with important immunomodulatory, antitumor, antiviral, and growth inhibitory properties. Consistent with the multiplicity of their effects, the IFNs activate several cellular cascades after they bind to their receptors. Although the Jak-Stat pathway is the most studied pathway, it appears that additional signaling cascades play roles in IFN signaling. The phosphatidylinositol 3'-kinase (PI3'K) pathway has emerged as one of the critical players in IFN signal transduction and is the focus of this review. This pathway is activated distinctively from the Stat pathway but plays important roles in the regulation of IFN-dependent gene transcription and initiation of mRNA translation. PMID- 16375607 TI - Alternative and accessory pathways in the regulation of IFN-beta-mediated gene expression. AB - Type I interferons (IFNs) induce the transcription of IFN-stimulated genes (ISGs) through activation of the Jak-Stat pathway. Although some determinants of specificity are dictated by the Jak-Stat components, recent observations indicate that the system incorporates other components for selectivity and flexibility, whose mechanisms remain to be defined. We identified a gene, beta-R1, which was induced relatively selectively by IFN-beta as compared with numerous IFN-alpha subtypes. Because all type I IFNs equally activate Jak-Stat signaling to IFN stimulated gene factor 3 (ISGF3), this observation implied the existence of accessory signals for IFN-induced gene expression. We have used beta-R1 as a model system to examine this accessory signaling. In addition to Jak-Stat signaling for mediating IFN-induced cellular responses, p38 mitogen-activated protein kinase (p38 MAPK), phosphoinositol 3-kinase (PI3K), the IkappaB kinases (IKKs), and nuclear factor-kappaB (NF-kappaB) are some of the accessory components identified as required for the induction of certain IFN-beta-induced genes. This review focuses on the roles of accessory components in IFN-beta mediated signaling, mechanisms of accessory signal generation, and how they modulate gene induction. PMID- 16375608 TI - Alternative signaling pathways regulating type I interferon-induced apoptosis. AB - Type I interferons (IFNs) are pleiotropic cytokines that exert multiple effects on normal and tumor cells. These effects are supposedly mediated through the stimulation of several signal transduction pathways by type I IFNs. These include the well-studied canonical Jak-Stat pathway, largely responsible for the antiviral and growth-inhibitory activities of IFNs, as well as the phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase (MAPK) pathways, whose importance in IFN-induced biologic outcomes has not been precisely established. One of the effects of type I IFNs on tumor cells is the induction of programmed cell death, apoptosis, which has been studied extensively over the last decade and has been suggested to be an important effector mechanism for IFN's antitumor effects in the treatment of cancer. The aim of this review is to summarize and discuss the recent data in the field of type I IFN-induced apoptosis, with special emphasis on the molecular mechanisms of apoptosis and on the role of alternative noncanonical signaling pathways stimulated by type I IFNs in this process. PMID- 16375610 TI - Factors associated with sustained virologic suppression in patients receiving antiretroviral therapy in an urban HIV care clinic. AB - Clinical trials commonly measure rates of virologic suppression at a specific time point, whereas sustained virologic suppression is the goal of highly active antiretroviral therapy (HAART). We explored factors associated with sustained virologic suppression in an urban clinic population. The study population was drawn from patients who enrolled in Montefiore Medical Center's Infectious Diseases Clinic from 1999 to 2000. A computerized query of the hospital information system generated a list of potential case patients having every HIV- 1 viral load (VL) less than 50 copies per milliliter throughout 2002 (at least three VL measurements were required), and a list of potential controls who failed to demonstrate sustained virologic suppression during 2002. Demographic and clinical information were collected by chart review, and case and control patient characteristics were compared by both univariate and multivariate analyses. Sixty four case patients were compared to 64 controls. There were no significant differences in age, gender, ethnicity, type of antiretroviral therapy, or frequency of clinic visits. During the year of the study, cases experienced a significantly greater rise in CD4(+) lymphocyte counts than controls (108 cells per microliter versus 27 cells per microliter). On univariate analysis, factors associated with sustained virologic suppression included risk behavior other than heterosexual contact or injection drug use, being a non-smoker, and hepatitis C seropositivity. On logistic regression analysis, factors independently associated with sustained virologic suppression were risk behavior other than heterosexual contact or injection drug use (IDU), and hepatitis C seropositivity. In this study sample, being a nonsmoker, having a risk behavior for HIV acquisition other than heterosexual contact or IDU, and being seropositive for hepatitis C were associated with sustained virologic suppression. Computerized query of the hospital information system proved to be a powerful tool for the identification of study patients in a real-world clinic environment. PMID- 16375611 TI - The importance of potency and durability in HIV patient antiretroviral therapy preferences: a telephone survey. AB - Patients who were receiving or had received antiretroviral therapy (ART) participated in 45-minute telephone interviews to evaluate the importance of major treatment attributes. A Likert scale was used to quantify and rate the importance of 9 ART attributes. Trade-off exercises allowed participants to select a preferred hypothetical ART regimen from two options based on daily dosing and varied efficacy. Participants were asked to assume that all else about the medications was the same. A total of 387 patients were surveyed (72% male; 44% African American, 41% Hispanic; 28% with no high school diploma, 29% high school graduate, 25% college with no degree; 46% infected through men who have sex with men [MSM], 19% infected through injection drug use [IDU]). Efficacy attributes (lowering viral load, raising CD4, durability) were rated as "most important" or "very important" by significantly more patients than other attributes (resistance profile, appearance side effects, gastrointestinal side effects, dosing frequency, pill burden, cholesterol side effects). Similar results were seen for subgroups analyzed by gender, ethnicity, age, line of therapy, region, and route of infection. In the second set of questions, 92% of patients preferred more effective twice-daily regimens over less effective once daily regimens, and 89% preferred more durable twice-daily regimens over less durable once-daily regimens. Results suggest that potency, immune improvement, and durability of ART regimens are more important to HIV patients than other attributes such as side effects, dosing frequency, or pill burden. These results, in conjunction with other studies, suggest that patients prioritize viral suppression, immune improvement, and regimen durability more highly than regimen convenience. PMID- 16375612 TI - Antiretroviral medication errors in a national medication error database. AB - Antiretroviral medications have reduced the morbidity and mortality associated with HIV. In contrast to these benefits, medication errors involving antiretrovirals represent opportunities for causing harm. We analyzed 400 medication errors that involved at least one single or combined HIV antiretroviral product that had been reported to a national medication error reporting program (MEDMARX). Our analysis revealed that 3% of the errors were harmful. Most of the errors (45%) occurred in the dispensing phase of the medication use process, a finding that differs significantly from many published studies. The most frequent types of errors were wrong dose (37.5%) and wrong medication (32%). Lamivudine (Epivir, Glaxo-SmithKline, Research Triangle Park, NC) was the most commonly identified product to be involved in the errors. Community hospitals were more likely to have prescribing errors than teaching hospitals. Similar brand and generic names were associated with many of the errors. With frequent dosing of many HIV medications, health care organizations must have a process to clarify orders rapidly and maintain current references (including photos) of antiretrovirals. Prescribers should clearly spell out the intended product and avoid abbreviations. Participating in voluntary medication error reporting programs increases the awareness of the threats of medication errors involving antiretrovirals. PMID- 16375613 TI - Efficacy of antidepressant medication among HIV-positive individuals with depression: a systematic review and meta-analysis. AB - Depression is highly prevalent among HIV-positive individuals yet some, but not all, have not found antidepressant medication to be efficacious in this population. We performed a systematic review and meta-analysis of double-blinded, randomized controlled trials to examine efficacy of antidepressant treatment among HIV-positive depressed individuals and evaluate whether the results are generalizable to women and minorities. We used PubMed, the Cochrane Database, a search of bibliographies, and consultation with experts to find double-blinded, controlled clinical trials with random assignment to antidepressants or control condition for which HIV-positive patients met standard diagnostic criteria for depression. The principal measure of effect size was the standard difference between means on the Hamilton Depression Score (HDS). We identified 7 studies that included 494 subjects. Three of the 7 studies reported significant antidepressant effects. The pooled effect size from the random effects model was 0.57 (95% confidence interval [CI]: [0.28-0.85]). Heterogeneity across studies was significant (Q, 13.22; p = 0.07; I(2) = 47.1%). When stratified by placebo response, the pooled effect size for placebo response greater than 33% was 0.20 ( 0.11-0.52) and not significant while the pooled effect size for placebo response greater than 33% was 0.80 (0.52-1.08) and was significant. Placebo response explained nearly 62% of the variance in effect sizes across studies. Women were nearly absent from and minorities were underrepresented in the studies investigated. Antidepressant medication is efficacious in treating depression among depressed, HIV-positive individuals. However, the underrepresentation of women and minorities limits the generalizability of these findings and suggests that future studies be directed to address this disparity. PMID- 16375614 TI - Methods of HIV disclosure by men who have sex with men to casual sexual partners. AB - The purpose of this paper is to report results of a qualitative investigation into the methods that HIV-positive men who have sex with men (MSM) use to disclose their serostatus to casual sexual partners. In-depth, qualitative interviews were conducted with 57 HIV-positive adult MSM from a large midwestern city. Using an inductive approach to data analysis, our participants revealed a typology of disclosure consisting of strategies that can be placed into five primary categorizations of disclosure. These include: point-blank disclosure, indirect disclosure, stage setting, buffering, and seeking similars. This information can be especially beneficial to those working with men to develop better disclosure skills. PMID- 16375615 TI - Pillboxes and antiretroviral adherence: prevalence of use, perceived benefits, and implications for electronic medication monitoring devices. AB - HIV treatment adherence is improved by behavioral interventions, including medication organizers such as pillboxes, which can also interfere with adherence electronic monitoring devices (EMD). This study examined the characteristics of pillbox users and the implications of pillbox use for research using EMD adherence research. Men and women (n = 160) currently taking antiretroviral medications (ARVs) completed anonymous surveys in a community research setting. Sixty-three (39%) of individuals taking ARVs were currently using a pillbox. Pillbox users were significantly more likely to have an undetectable viral load and less likely to have missed their medications within a day of the assessment. Pillbox users indicated feeling that they benefited from using their pillbox and 76% reported they would likely stop using their pillbox if required to participate in a research study that paid them. Adherence EMD pose methodological limitations to research when pillbox users are excluded and ethical implications when pillbox users stop using their pillbox to participate. PMID- 16375616 TI - People with HIV as educators of health professionals. AB - This qualitative study examined the impact upon people living with HIV (PHAs), of being trained and utilized as educators of health professionals. PHAs participated in a training program to help them develop skills to facilitate learners in problem-based educational events. After training the PHAs participated in small group problem-based tutorials with separate groups of physiotherapy and occupational therapy students and family medicine residents. Content analyses of the PHAs' reflective journals and semistructured interviews conducted at completion of the project indicated there was a positive impact on their teaching skills, self-awareness, personal understanding of HIV, confidence in teaching, and everyday life. Learner feedback indicated that they valued their interactions with the PHAs. This model of education has the potential to positively benefit patients living with a variety of illnesses and disabilities. PMID- 16375617 TI - Sexually transmitted disease/HIV and heterosexual risk among miners in townships of Yunnan Province, China. AB - The HIV epidemics in some areas of Yunnan Province, southwestern China, are close to becoming generalized, demanding the need for a better understanding of sexually transmitted disease (STD) and heterosexual HIV risk. While female heterosexual risk is captured by sentinel surveillance, less is known about clients of commercial sex workers and other subsets of men at increased risk of STD/HIV. A convenience sample of 232 miners in townships of Yunnan Province completed a questionnaire and provided specimens for STD/HIV testing. Relative prevalences of HIV, gonorrhea, and chlamydia among sexually experienced miners surveyed were 0.5% (1/182), 0.5% (1/182), and 9.3% (17/182), respectively. Chlamydia seropositivity was significantly associated with receiving less education (p = 0.03). Among all respondents, 9.4% admitted to purchasing sex, 82.9% had not used a condom in the last 6 months, and substantial gaps in knowledge about STD/HIV were apparent. Because preventing heterosexual spread of HIV is crucial in this area of China to prevent a generalized epidemic, better understanding and designing low literacy interventions targeted at sex worker clients and similar subsets of high-risk migrants may be warranted. PMID- 16375618 TI - Sexually transmitted infections among female sex workers in Yunnan, China. AB - A cross-sectional prevalence survey of sexually transmitted infections (STIs) was conducted among female sex workers (FSWs) in Kunming, Yunnan Province, China. A total of 505 FSWs participated in the survey. All eligible participants gave informed consent. Demographic, behavioral, and clinical information of the participants was gathered by direct structured interviews. Tampon swabs were collected to test for Chlamydia trachomatis, Nesseria gonorrhoeae, and Trichomonas vaginalis and serum specimens were collected to test for HIV antibody by enzyme-linked immunosorbent assay (ELISA) screening and Western blot confirmation, syphilis with rapid plasma reagin (RPR) and T. pallidum hemagglutination (TPHA) confirmation, and for HSV-2 antibodies with the HerpeSelect 2 ELISA. The most prevalent bacterial STI was Chlamydia trachomatis (58.6%), followed by Trichomonas vaginalis (43.2%) and Neisseria gonorrhoeae (37.8%). Three hundred twenty-seven (65.1%) sex workers had serologic evidence of exposure to HSV-2 infection. Ten percent were positive for HIV infection, all injecting drug users (IDUs); and 9.5% had infection with syphilis. Comprehensive prevention and effective STI services for sex workers and their clients will be the key strategies to the control of STIs, including HIV. Policies and prevention strategies for STI/HIV need to focus on high-risk subpopulations, such as sex workers (particularly sex workers who inject drugs) and their clients. PMID- 16375621 TI - Fixing broken heart valves percutaneously: the shape of things to come? PMID- 16375623 TI - Clopidogrel use in coronary artery disease. AB - Platelet adhesion, activation and aggregation are central to the pathophysiology of the acute coronary syndromes. Clopidogrel, an oral thienopyridine derivative, is a platelet adenosine diphosphate receptor antagonist capable of inhibiting platelet activation. During the last decade, the utility of clopidogrel has been evaluated in several common clinical scenarios in a large number of patients. The benefits of clopidogrel in patients with stable coronary artery disease undergoing elective percutaneous coronary interventions and in patients presenting with acute coronary syndromes are now well established. This review outlines the pharmacology of clopidogrel, highlights the results of clopidogrel trials in the setting of acute coronary syndromes, and presents areas of uncertainty and potential future work. PMID- 16375624 TI - Conivaptan: a selective vasopressin antagonist for the treatment of heart failure. AB - Heart failure commonly manifests as a syndrome of salt and water retention. Arginine vasopressin plays an important role in volume homeostasis and may contribute to this syndrome seen in heart failure patients. Recently, a number of agents have been developed that antagonize the effects of vasopressin. Conivaptan, which is a dual antagonist of the V1a and V2 receptor, has shown promise in animal studies and in small scale human trials as a potential therapeutic option for the treatment of acute and chronic heart failure. Further large studies are being conducted, which may confirm the benefits of conivaptan and other vasopressin antagonists in heart failure patients. PMID- 16375625 TI - Aortic stenosis and the failing heart. AB - The combination of aortic stenosis and left-ventricular dysfunction is a challenging situation for the physician. Diagnosis of this condition requires a detailed evaluation to understand the etiology and reversibility of the ventricular dysfunction and to accurately determine the real severity of the stenosis. Whether the aortic stenosis the cause of the left ventricular failure or is an independent disease has significant diagnostic, prognostic and therapeutic implications. Dobutamine echocardiography provides critical information to determine the real severity and the left ventricle's potential to recover (contractile reserve). Attempts to delay the progression of the aortic stenosis with medical treatment have been limited, and valve replacement remains the hallmark of ultimate treatment. If surgery is inadvertently delayed, left ventricular systolic dysfunction will result in clinically evident congestive heart failure and this situation carries a very high short-term mortality. Aortic valve replacement in this setting improves the outcome, but perioperative mortality is high, and particularly when coronary revascularization is also needed, there is no ventricular contractile reserve and transvalvular gradients are low. Adequate timing of surgery is extremely important and increasingly more difficult. Management decisions should be tailored by the results of dobutamine echocardiography and made on a case-by-case basis. PMID- 16375626 TI - Heart failure therapy in children. AB - The most common reason for heart failure in children is volume overload secondary to a left-to-right shunt. Therefore, an accurate diagnosis with identification of possible surgical or interventional reactions should be the first priority. Medical therapy is mainly based on diuretics, angiotensin-converting enzyme inhibitors, cardiac glycosides and beta-blockers. There are few prospective trials in pediatric cardiology, but the available data reach a similar conclusion to that of adults with heart failure. Diuretics are an important tool in patients with fluid retention, and angiotensin-converting enzyme inhibitors are helpful in patients with volume overload of the ventricles. Cardiac glycosides are still in use, but there is a trend toward primary use of diuretics. Angiotensin-converting enzyme inhibitors and beta-blockers have been used successfully in the treatment of heart failure in children, but there are limited data on its efficacy. PMID- 16375627 TI - The role of erythropoietin in myocardial protection: potential mechanisms and applications. AB - The glycoprotein erythropoietin was originally discovered as a principal regulator that promotes the survival, proliferation and differentiation of erythroid progenitor cells. Despite potentially detrimental effects, such as increased blood pressure and hyperviscosity, recombinant human erythropoietin has been demonstrated to be a safe drug, as millions of anemia sufferers have received it over the last decade as a form of treatment. Recently, erythropoietin receptors have been discovered in a variety of tissues, including the cardiovascular system, and erythropoietin has been demonstrated to have a beneficial effect in congestive heart failure patients with anemia. The purpose of this review is to summarize the pleiotropic cardioprotective effects of erythropoietin in the cardiovascular system and to evaluate its potential role as a biomarker in these disorders. PMID- 16375628 TI - The prognostic value of anemia, right-heart catheterization and neurohormones in chronic heart failure. AB - Chronic heart failure is increasing in incidence and prevalence. Recent advances in medical therapy have improved prognosis such that, even in patients with chronic heart failure who are New York Heart Association Classes III and IV, annual mortality can be as low as 11.4%. Nevertheless, some patients remain at risk, despite optimal disease-modifying medical therapy, and it would seem appropriate that these patients are considered first for appropriate device therapy or for the scarce resource of cardiac transplantation. Many parameters have been assessed for their prognostic potential in patients with chronic heart failure. In this review, pertinent studies investigating anemia, right-heart hemodynamics and neurohormones as prognostic markers are discussed. PMID- 16375629 TI - Imaging techniques in cardiac electrophysiology. AB - Modern cardiac electrophysiology procedures include catheter-based arrhythmia ablation and transvenous device implantation, which are highly dependent on accurate, real-time cardiac imaging. With the realization that anatomic structures are critical to successful electrophysiologic procedures, accurately defining a patient's cardiac anatomy has become more important. Fluoroscopy allows for 2D imaging of cardiac structures in real-time, and is used to guide catheter and lead placement, but does not allow for visualization of soft tissues. Intracardiac echocardiography allows for both direct visualization of anatomic structures within the heart and real-time imaging during catheter placement. Despite advances in intracardiac echocardiography catheters that allow for larger windows, the ability to accurately delineate anatomic structures depends on the patient's anatomy and operator experience. Neither of these techniques allows for electrical mapping of the heart; however, both anatomic and electrical intracardiac mapping can be achieved with advanced mapping systems. These systems allow for real-time catheter localization, help elucidate cardiac anatomy, evaluate electrical activation during arrhythmias and guide catheter placement for deliverance of radiofrequency current. More recently, 3D cardiac computed tomography has been used to accurately define intracardiac anatomy; however, catheter tracking and electrical mapping cannot be performed by computed tomography. Mapping systems are now being merged with computed tomography images to produce an accurate anatomic and electrical map of the heart to guide catheter ablations. The objective of this paper is to describe the current imaging and mapping techniques used in electrophysiologic procedures. PMID- 16375630 TI - Methods of cerebral protection in surgery of the thoracic aorta. AB - During the last decade, a considerable increase in the number of operations on the thoracic aorta has been observed. Although patient's outcomes have improved considerably, this surgery is still associated with significant morbidity and mortality due to neurological complications. Various methods have been proposed and widely used as means to protect the brain from ischemic damage. This review summarizes the principal methods of cerebral protection, describes the advantages and disadvantages of each method and their impact on patient outcomes, and discusses the different surgical techniques proposed to minimize the risk of cerebral injuries. PMID- 16375631 TI - Arrhythmias in adult congenital heart disease. AB - Recent advances in pediatric cardiology and cardiac surgery have allowed a rapidly expanding population of patients with congenital heart disease to thrive well into their adult years. Often after prolonged uneventful clinical courses as children and adolescents, arrhythmias may surface later in life. These arrhythmias are a major source of morbidity. In addition, sudden death is the leading cause of mortality in adult patients with congenital heart disease. This review highlights the various types of brady- and tachyarrhythmias encountered in the more common forms of adult congenital heart disease and explores prognostic implications and therapeutic options. PMID- 16375632 TI - Biventricular pacing in heart failure: a review. AB - Biventricular pacing has been an exciting recent advance in the management of drug-refractory heart failure. This new therapy has evolved as much from necessity as scientific observation, since benefits derived from pharmacotherapy currently appear to have reached their peak. Clinical trials of biventricular pacing are establishing morbidity and mortality benefits in heart failure. New challenges in the use of these pacemakers are now arising. These include the accurate diagnosis of ventricular dyssynchrony and, hence, potential responders to the refinement of implantation of the left ventricular lead to the appropriate dyssynchronous ventricular area and optimization of pacemaker programming. This review gives a general overview of the principles and the current evidence for the use of biventricular pacemakers in the treatment of heart failure. In addition, a discussion of current research and future projects is included. PMID- 16375633 TI - Prehypertension and risk of cardiovascular disease. AB - Epidemiologic data have established a continuous relationship between vascular risk and blood pressure that extends down to levels as low as 115/75 mmHg, emphasizing the lack of a critical threshold value that defines 'high' blood pressure. Acknowledging the graded and continuous nature of the relations of blood pressure to vascular risk, the Seventh Report of the Joint National Committee on the Prevention, Detection, Evaluation and Treatment of High Blood Pressure (JNC VII) introduced the new category 'prehypertension' to describe people with a systolic blood pressure between 120 and 139 mmHg and/or a diastolic blood pressure between 80 and 89 mmHg. It is estimated that 31% of the US population (70 million) has prehypertension. The risk of cardiovascular disease within this large prehypertensive population is not uniform, however, and increases with a rising concomitant burden of other vascular risk factors. Accordingly, a strategy of estimating global cardiovascular risk (by applying standardized risk prediction algorithms) and adjusting the intensity of blood pressure lowering (and reduction of other risk factors) to the absolute risk of cardiovascular disease is desirable in prehypertensive individuals. Adopting a healthier lifestyle, as recommended by JNC VII, is a critical component of the therapeutic approach to prehypertension. PMID- 16375634 TI - Chagas cardiomyopathy and ischemic stroke. AB - American trypanosomiasis, known as Chagas disease is a major cause of cardiomyopathy in South America. Irreversible damage to the heart can appear 10 20 years after chagasic infection. The relationship between Chagas cardiomyopathy and ischemic stroke has been reviewed. Significant variables that predict ischemic stroke in chagasic patients have been identified: apical aneurysm, cardiac insufficiency, electrocardiogram arrhythmia and female gender. Chagasic cardiomyopathy should be included in the differential diagnosis of the etiology of stroke, being a potential source of cardioembolic stroke. PMID- 16375635 TI - The role of infection and immunity in atherosclerosis. AB - Cardiovascular diseases account for 20% of deaths worldwide, rising to 50% in developed countries. Current understanding of atherosclerosis derives from a combination of research in animals and cell cultures, analysis of human lesions, clinical investigations of patients with acute coronary syndromes and epidemiological studies of coronary artery disease. By measuring serologic titers in the serum of patients after cardiovascular events, it was observed that the greater the infectious exposure of a patient, the larger the atherosclerotic lesion extension. In addition, gene targeting or pharmacological inhibition of certain cytokines aggravates atherosclerosis in animal experiments. Other animal experiments have succeeded in proving that B cells play a protective role in atherosclerosis through induced immunity against oxidized low-density lipoprotein and other epitopes. Molecular mimicry might respond to the question of how infection may trigger vulnerability in previously stable atherosclerotic lesions. The FLU Vaccination Acute Coronary Syndromes trial enhanced the debate on atherosclerosis prevention by the application of antiflu vaccine. So far, antibiotics have failed to reduce cardiovascular risk, as recent trials could not demonstrate a statistically significant risk reduction. Having assumed atherosclerosis to be an inflammatory disease, the WHO considered the possible role of secondary prevention with antiflu vaccine. PMID- 16375638 TI - Letrozole as adjuvant endocrine therapy in postmenopausal women with breast cancer. AB - The third-generation aromatase inhibitor letrozole offers a promising approach to treating hormone-sensitive breast cancer for postmenopausal women, through potent and specific inhibition of estrogen synthesis. In neoadjuvant and first-line treatment, letrozole demonstrated superior efficacy compared with tamoxifen in randomized Phase III trials. Initial results of Breast InterGroup 1-98, a large ongoing randomized trial investigating primary adjuvant endocrine treatment with either letrozole or tamoxifen, have recently been presented. Patients treated with letrozole demonstrated a 19% improvement in disease-free survival and a significant reduced risk of distant recurrences, holding out the prospect of a survival advantage over tamoxifen treatment with further maturation of the trial. For patients who have already completed 5 years of tamoxifen, extended endocrine therapy with letrozole is a new therapeutic option based on the results of the MA 17 trial. The optimal use of aromatase inhibitors remains an open question, at least until results from randomized trials (BIG 1-98, TEAM) investigating the sequential use of an aromatase inhibitor and tamoxifen in comparison with continuous monotherapy become available. PMID- 16375639 TI - Robotic prostatectomy: a pooled analysis of published literature. AB - OBJECTIVE: Robotic prostatectomy has been recently added to the treatment armamentarium of localized prostate cancer. We reviewed published data on this modality with critical appraisal of outcomes and complications. METHODS: A Medline search was performed that encompassed all published articles on robotic prostatectomy. Select data on open radical retropubic (RRP) and laparoscopic (LP) prostatectomy from centers of excellence in the USA and Europe were included for objective comparison. Perioperative parameters, pathological results, including surgical margin rate, complications and postoperative continence and potency rates were reported. Advantages and shortcomings of robotic prostatectomy, and the learning curve and cost issues were also addressed. RESULTS: Robotic prostatectomy offers advantages of minimally invasive access surgery, including enhanced visualization, decreased bleeding and transfusion rate, shorter hospital stay and faster recovery. Pathological outcomes are comparable to RRP and LP with acceptable positive margin rate. At short-term follow-up, continence and potency results appear to be equivalent to RRP and LP. The learning curve of robotic prostatectomy is faster than that of LP. Cost remains a matter of debate at the present time. Long-term cancer control results of robotic prostatectomy are still maturing. CONCLUSIONS: Robotic prostatectomy is a promising minimally invasive surgical approach for men with localized prostate cancer. Short-term clinical and pathological results compare favorably to RRP and LP. PMID- 16375640 TI - Benefits of laparoscopic prostate-sparing radical cystectomy. AB - PURPOSE: Prostate-sparing radical cystectomy has been described in the literature and has proven to be a promising procedure owing to the continence and erectile function results without necessarily compromising the oncologic outcome in selected patients. TECHNIQUE POINTS: A transperitoneal approach is used. Lymph node dissection is performed with frozen section, and then the ureters are ligated and biopsied. The seminal vesicles are dissected, followed by complete mobilization of the bladder. Incision of the bladder neck is performed, followed by simple prostatectomy. Finally, bowel reconstruction is carried out via a small infra-umbilical incision that also permits the extraction of the surgical specimen and the anastomosis of neobladder to the prostate capsule. A total of 25 patients have undergone this procedure, with average surgical times of 285 min and blood loss of 640 ml. The complications encountered included: one bowel incarceration, one urinary leak, one lymphocele and one port hernia. At median 9 months follow-up, no patient presented with daytime incontinence, although seven complained of nocturia. A total of 20 patients maintained their preoperative sexual potency, and four reported a postoperative decrease in their erectile function. CONCLUSIONS: Laparoscopic prostate-sparing radical cystectomy appears to be oncologically safe, reproducible and has promising functional benefits. The authors believe this procedure presents a good option in very select patients. PMID- 16375641 TI - Fertility-sparing therapy for young women with endometrial cancer. AB - Approximately 1600 women under the age of 40 will be diagnosed with endometrial cancer this year. For these women, preservation of fertility may be one of the most important factors when seeking treatment options. The objective of this article is to review the available literature on fertility-sparing therapy for young women with endometrial cancer. Patient selection, pretreatment testing, treatment regimens, response rates and pregnancy outcomes will be explored. Recommendations regarding this approach for the treatment of endometrial cancer in young women will also be made. PMID- 16375642 TI - An overview of uterine cancer and its management. AB - Endometrial cancer is increasingly common in affluent Western countries, largely owing to the growing obesity of those populations. There are two recognized types of endometrial cancer: Type I is more common and is associated with obese postmenopausal women and comprises approximately 80% of all endometrial cancers; Type II describes a woman who is often younger and thinner with a more aggressive histologic type that is nonestrogen dependent, of either serous or clear cell histology, and consists of a more aggressive clinical course and results in poorer prognosis. As the majority of patients with endometrial cancer present with symptoms and have early disease, screening is unlikely to be cost effective or reduce the mortality rate. However, surveillance of high-risk populations is a different proposition. Patients who may benefit from routine surveillance include those with a family history of endometrial cancer, a history of hormone replacement therapy with less than 12-14 days of progestogens, long-term use of tamoxifen, hereditary nonpolyposis colorectal cancer family syndrome, Cowden's syndrome, Peutz-Jeghers syndrome, a history of breast cancer and obesity. Most patients with endometrial cancer are offered surgery as first-line therapy. The standard surgical procedure should be an extrafascial total hysterectomy with bilateral salpingo-oophorectomy. Adnexal removal is also recommended, even if the adnexa appear normal, as they may contain micrometastases. The safety of a laparoscopic approach in the surgical management of uterine cancer has not yet been demonstrated in prospective randomized trials, therefore, the field awaits the Gynaecologic Oncology Group's prospective Lap-2 study. While post-treatment follow-up guidelines vary between institutions and countries, in general, patients at high risk of recurrence are followed closely every 3-4 months for the first year or two, then every 6 months to complete 5 years of follow-up. PMID- 16375643 TI - Hormonal therapy in epithelial ovarian cancer. AB - The ovary is an endocrine and end organ. Hormones and their receptors have been associated with ovarian cancer and may be related to its causation. Some data suggest that hormonal therapies may have an effect on ovarian cancer in palliative settings. The most well studied anticancer drugs are tamoxifen, megestrol acetate, medroxyprogesterone acetate, leuprolide acetate, anastrozole and letrozole. Presently, no hormonal therapy is approved by the US FDA for the treatment of any type of ovarian malignancy or is listed as an active agent by any of the authoritative compendia. Owing to the endocrine associations with ovarian cancer, the minimal side effects of hormonal therapy and the demonstrated activity of hormonal therapies in other endocrine organ-associated malignancies, further study of hormonal therapies for ovarian cancer is warranted. PMID- 16375644 TI - Risk of second cancers after radiotherapy for cervical cancer. AB - Radiotherapy for cervical cancer has both beneficial and detrimental effects: improvement of patient survival and potential induction of a second cancer among long-term survivors. Large epidemiological studies have demonstrated small, but significant, increases of second cancers with radiotherapy compared with the general population. The risk of second cancer has been characterized by organ sites, dose, time since radiotherapy and age at the time of radiotherapy. Analyses of genetic susceptibility and molecular carcinogenesis can be used to develop more appropriate strategies for radiation therapy for cervical cancers. PMID- 16375645 TI - Current and future status of prostate cancer chemoprevention. AB - Prostate cancer remains one of the most common malignancies in Western countries and is also increasing in Asian countries. The high incidence, influence of environmental factors, such as androgen, and extended natural history of prostate cancer provide multiple opportunities for prevention; therefore prostate cancer is an ideal candidate for chemoprevention. The development of chemoprevention strategies against prostate cancer would have a huge impact, both medically and economically. Large-scale clinical trials suggest that some agents can prevent the development of prostate cancer. Active research into chemoprevention is ongoing and gathered evidence will provide effective and safe strategies to prevent prostate cancer. This review will highlight the present status of prostate cancer chemoprevention and other mechanisms of prevention, as well as discussing the issues to be addressed. PMID- 16375646 TI - The endothelin axis in urologic tumors: mechanisms of tumor biology and therapeutic implications. AB - Endothelin (ET)-1 and its receptors ET-A and ET-B, referred to commonly as the endothelin axis, have been identified in various human cancers, especially gynecologic tumors, such as breast cancer or ovarian cancer, but also including urologic tumor entities. They play a key role in tumor growth and progression by influencing critical cancer pathways, such as apoptosis, angiogenesis and proliferation. In prostate cancer, overexpression of the ET-A receptor increases with tumor progression, and clinical trials with selective ET-A receptor antagonists, such as atrasentan (ABT-627), have shown promising early results. In preclinical models of bladder cancer, overexpression of the ET axis has been demonstrated and ET-targeting agents are under investigation. This paper reviews the role of the ET axis in human cancers and focuses on preclinical and clinical studies in urologic tumor entities to further define the role of ET-targeting agents as targeted molecular therapy. PMID- 16375647 TI - Developing innovative strategies for advanced transitional cell carcinoma of the bladder. AB - Several improvements in the treatment of advanced transitional cell malignancies have been provided by clinical trials in the past 10 years. Nonetheless, there are conflicting results regarding the effect of perioperative chemotherapy of muscle-invasive disease and new cytotoxic agents in the metastatic setting. The authors will discuss the results of major clinical trials and examine developing targeted-oriented treatment strategies. PMID- 16375648 TI - Aurora A and B kinases as targets for cancer: will they be selective for tumors? AB - Aurora A and B kinases are closely related kinases involved in regulating separate points in the cell cycle. This review highlights the rationale for Aurora kinases as cancer targets and examines the currently known Aurora kinase inhibitors in the patent and scientific literature. The known crystal structures of the Aurora kinases are described with relevance to bound ligand interactions and the prospect of the generation of drug-resistant mutant forms. The potential for selectivity versus primary cells will also be discussed. The status of the inhibitors in clinical development is described. PMID- 16375649 TI - Chemotherapy for metastatic melanoma. AB - Despite the limited efficacy of systemic chemotherapy in the treatment of metastatic melanoma, it remains the gold standard in the case of patients with a good performance status and no major comorbidities for whom radical surgery is unsuitable. Various drugs have been employed as monochemotherapy with response rates ranging from 0 to 20%. Many Phase III trials have compared the role of polychemotherapy with that of single-agent chemotherapy, or evaluated the impact of biological response modifiers alone or in combination with chemotherapeutic agents. However, the current scenario does not seem to be significantly different from the situation of 20 or 30 years ago. To date, no single drug, combination chemotherapy in addition to a hormonal or biotherapy compound, has demonstrated an overall survival benefit in a randomized clinical trial. PMID- 16375650 TI - From Rapa Nui to rapamycin: targeting PI3K/Akt/mTOR for cancer therapy. AB - One of the most prominent pathways explored in the area of targeted therapy is the PI3K/Akt/mTOR pathway, which plays a central role in cell survival and proliferation. Deregulation of this pathway has been implicated in the promotion of cancer cell growth and survival. Inhibition of several steps of this pathway has been shown to confer favorable antitumor activity in a variety of cancer types. This article provides a brief analysis of the PI3K/Akt/mTOR pathway, its importance in tumor pathogenesis and the current status of preclinical and clinical studies targeting signaling components of this pathway. PMID- 16375651 TI - Maintenance biotherapy with interleukin-2 and interferon for metastatic renal cell cancer. AB - The term maintenance immunotherapy comprises at least two different therapeutic approaches: the continuation of immunotherapy beyond disease progression and the use of chronic immunotherapy after the achievement of an initial response (or disease stabilization) with more intensive treatment modalities, such as chemotherapy. The former therapeutic approach was proposed in renal cell carcinoma some years ago relying on several immunological considerations. Some years later, we have learned that it is feasible and endowed with a favorable therapeutic index; furthermore, its immunologic effects are well described and reproducible, and it has antitumor activity. However, due to the lack of adequate randomized Phase III studies, the actual impact of this treatment strategy on patient survival has not yet been proved. The rationale of this treatment, its immunological and clinical results, as well as its pitfalls and perspectives, will be presented and discussed in this review. PMID- 16375652 TI - Advances of Alzheimer's disease research: crossroad of basic and translational studies. PMID- 16375653 TI - Activity-dependent synapse modulation and the pathogenesis of Alzheimer disease. AB - During normal development of the nervous system, a major reduction occurs in the initially excessive number of neurons and synapses. This "pruning" process is heavily influenced by patterns of electrical activity in the synaptic circuits being pruned. Many of the cell biological and molecular mechanisms involved in this activity-dependent modification of nervous system structure and function have been explicated, and the area is one of intense study. Similarly, an explosive increase has occurred in knowledge about the molecular pathogenesis of Alzheimer disease (AD). There are significant mechanistic commonalities between the normal neurodevelopmental process and development of AD. We hypothesize that abnormalities in neural activity patterns, or in the coupling between neural activity and maintenance of neurons and synaptic circuits, may be a key determinant in the pathogenesis of AD that is late in onset, sporadic in nature, and in which the genes for the presenilins and the beta amyloid precursor protein are normal. Behavioral data suggests that an active, socially engaged life-style may be associated with a reduced risk for AD. If so, mechanisms linking neural activity with synaptic circuit integrity are probably involved and provide a target for ameliorative pharmacological intervention. PMID- 16375654 TI - Recent insights on the pro-apoptotic phenotype elicited by presenilin 2 and its caspase and presenilinase-derived fragments. AB - Programmed cell death (PCD) also called apoptosis, is a normal and genetically controlled event that could play, when mis-regulated, a pivotal role in the development of several neurodegenerative disorders such as Parkinson's disease. Sporadic Alzheimer's disease is one of the most prominent age-related syndromes whose etiology, although still unknown, could be related to biochemical or environmental causes. A few cases of Alzheimer's disease are likely of genetic origin and linked to mutations on the genes coding for the amyloid precursor protein (betaAPP) and presenilins 1 and 2. Although still discussed, the hypothesis of an implication of apoptotic cell death in Alzheimer's disease neuropathology has been recently supported by a growing body of biochemical evidences. Thus, the implication of presenilins in apoptotic processes in vitro has been well documented but the mechanisms underlying this function are still a matter of intense research. The aim of this review is to focus on the mechanisms by which presenilin 2 affects the programmed cell death with special emphasis on the role of the proteolytically derived presenilin fragments generated by both presenilinase- and caspases. The distinct apoptotic phenotypes elicited by the two parent proteins presenilins 1 and 2 and their functional cross talk will be briefly discussed. PMID- 16375655 TI - Alzheimer's disease and P300: review and evaluation of task and modality. AB - Early stage Alzheimer disease patients and matched elderly unaffected controls (n = 16/group) were evaluated with the P300 event-related brain potential (ERP). All subjects performed four oddball tasks that varied systematically in task difficulty and were each presented in the auditory and visual modalities. P300 amplitude was smaller and peak latency longer for the Alzheimer patients compared to elderly control subjects across tasks and modalities. P300 differences between Alzheimer patients and controls were largest for the relatively easy tasks, with little influence of stimulus modality observed. The results suggest that the P300 brain potential is sensitive to Alzheimer's disease processes during its early stages, and that easily performed stimulus discrimination tasks are the clinically most useful. Theoretical and practical implications are reviewed. PMID- 16375656 TI - Cholesterol, copper and Abeta in controls, MCI, AD and the AD cholesterol lowering treatment trial (ADCLT). AB - Cholesterol clearly plays an influential role in promoting the production of amyloid beta (Abeta) and possibly the progression of Alzheimer's Disease (AD). The AD Cholesterol-Lowering Treatment trial (ADCLT; 1 year duration) tested atorvastatin and found significant benefit on measures of cognition and depressive symptoms in treated patients (N = 32) compared to placebo (N = 31). We assessed the circulating levels of Abeta(1-40), Abeta(1-42), ceruloplasmin (copper chaperone), apolipoprotein E and HDL-cholesterol in blood collected at each clinical visit during the ADCLT. We also determined the circulating cholesterol, ceruloplasmin, and Abeta levels in AD and MCI (mild cognitive impairment) patients, and controls (two groups stratified by function; high and low) participating in our Brain Bank Program. Each Brain Bank individual was clinically assessed for performance on the Mini-Mental Status Exam (MMSE), Rey auditory verbal learning test (AVLT), Clock draw, and UPSIT (smell identification test). Among individuals of equal age and education, scores on the MMSE were significantly reduced in AD compared to both MCI and controls, as were scores on the UPSIT. Ability on delayed verbal recall was significantly reduced in AD compared to MCI, and in MCI compared to both control groups. Performance on the Clock draw was similar for AD and MCI patients, but was significantly reduced when comparing MCI to control. Both cholesterol and ceruloplasmin levels were significantly increased in low-function controls compared to the high-function control group, but were not different from levels identified in the MCI and AD patients. Significantly increased levels of Abeta(1-40) occurred in low- compared to high-function controls, with a further significant increase in MCI compared to low-function controls. Circulating Abeta(1-40) levels were decreased in AD compared to MCI. Levels of Abeta(1-42) were not significantly different between the groups. The slight gradual increase in circulating Abeta(1-40) and Abeta(1 42) levels produced by atorvastatin treatment in the ADCLT were not significant compared placebo. There was a trend for significant reduction in circulating ceruloplasmin levels after a year of atorvastatin therapy compared to levels observed at screen. The levels of HDL-cholesterol remained stable in the atorvastatin treated AD patients for 9 months and then decreased significantly compared to the placebo group at the 1-year time-point. The combined data support a role for cholesterol in AD and a possible influence of increasing circulating copper levels. The deterioration of function in controls and transition to MCI may be associated with concomitant incremental increases in circulating Abeta(1 40) levels. Increased cholesterol and ceruloplasmin levels may be associated with slight deterioration in function among controls as a precursor to impairment considered MCI. The clinical benefit of atorvastatin therapy is clearly not associated with decreased circulating Abeta or increased HDL-cholesterol, but a positive influence of reduced copper (ceruloplasmin) levels may be a consideration. PMID- 16375657 TI - A randomized, double-blind, placebo-controlled trial of two doses of Ginkgo biloba extract in dementia of the Alzheimer's type. AB - CONTEXT: Previous studies of Ginkgo biloba extract (GbE) in patients with various forms of cognitive impairment or dementia have shown promising results. OBJECTIVE: To determine the clinical efficacy of GbE in mild to moderate dementia of the Alzheimer type. DESIGN: Randomized, placebo-controlled, double-blind, parallel-group, multicenter trial. SETTING: Outpatient clinics of universities and private research centers specialized in dementia. PATIENTS: 513 outpatients with uncomplicated dementia of the Alzheimer's type scoring 10 to 24 on the Mini Mental State Examination and less than 4 on the modified Hachinski Ischemic Score, free of other serious illnesses and not requiring continuous treatment with any psychoactive drug. INTERVENTION: 26-week treatment with GbE at daily doses of 120 mg or 240 mg or placebo. MAIN OUTCOMES: Cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-cog), Alzheimer's Disease Cooperative Study Clinical Global Impression of Change (ADCS-CGIC). RESULTS: There were no significant between-group differences for the whole sample. There was little cognitive and functional decline of the placebo-treated patients, however. For a subgroup of patients with neuropsychiatric symptoms there was a greater decline of placebo-treated patients and significantly better cognitive performance and global assessment scores for the patients on GbE. CONCLUSION: The trial did not show efficacy of GbE, however, the lack of decline of the placebo patients may have compromised the sensitivity of the trial to detect a treatment effect. Thus, the study remains inconclusive with respect to the efficacy of GbE. PMID- 16375658 TI - A randomized, double-blind, placebo-controlled pilot trial of safety and tolerability of two doses of divalproex sodium in outpatients with probable Alzheimer's disease. AB - OBJECTIVE: The Alzheimer's Disease Cooperative Study (ADCS) is conducting a clinical trial to address whether chronic valproate treatment can delay emergence of behavioral symptoms in outpatients with AD. Since there were no data on the safety and tolerability of divalproex sodium in outpatients with dementia, we undertook a pilot study to inform the design of the ADCS study. METHODS: We recruited 20 outpatients with probable AD, MMSE 10-20, without history of agitation or psychosis. This was a 10-week randomized, double-blind, placebo controlled study assessing the safety and tolerability of 1,000 mg/day and 1,500 mg/day of divalproex sodium delayed-release for 8 weeks followed by extended release for 2 weeks. Other outcome measures addressed cognition, function, global status, side effects, and laboratory data. RESULTS: Participants assigned to active treatment ingested approximately 30% less of their prescribed study medication compared to those receiving placebo (p < .05 Wilcoxon Rank Sum test). The average tolerated dose for all participants at week 8 was 810 mg/day or 11.5 mg/kg/day, similar to the dose tolerated by nursing home patients. The most common side effects were sleepiness and tiredness, with worse cognitive performance in those assigned to 1500 mg/day. CONCLUSIONS: These results were used to design the multi-center ADCS trial. Doses of less than 1000 mg/day of divalproex sodium were the maximum tolerated by these outpatients with AD. A larger study of divalproex sodium dose tolerability is needed to define treatment in outpatients with Alzheimer's disease. PMID- 16375659 TI - Induction of RhoGAP and pathological changes characteristic of Alzheimer's disease by UAHFEMF discharge in rat brain. AB - Novel experiments with Ultrasound Associated with High Frequency Electromagnetic Field (UAHFEMF) irradiation on rats and mice found evidences of characteristic Alzheimer's disease (AD) degenerations including neurite plaques, beta-amyloid, TAU plaque and deposition in cells, Neuro-Fibrillary Tangle and Paired Helical Filament (PHF) with rats and mice irradiated up to 2454 hours. Concomitant passive avoidance test was performed on six mice, and all showed signs of visual and auditory agnosia and lost cognition of threatening condition. The post section Thioflavin-S fluorescent microscopy found dilated ventricles and dense amyloid-deposition in Ca3 and dentate gyrus. In addition, PHF was identified in the 2454 hours-irradiated rat brain by electron microscope. A human T-cell activation RhoGTPase-activating protein (TAGAP) isoform b homolog (GenBank accession # P84107) induced in the UAHFEMF-treated rat brain was identified using electron spray ionization (ESI) liquid chromatography tandem mass spectrometry (LC/MS/MS). We hypothesized that one of the causes of AD can be the UAHFEMF discharges in human brain. PMID- 16375660 TI - Progression of mild cognitive impairment to Alzheimer's or vascular dementia versus normative aging among elderly Chinese. AB - To compare differences in evolutionary progressions from Mild Cognitive Impairment (MCI) to dementia of Alzheimer's type (DAT) or to vascular dementia (VaD) versus normal aging, subjects identified as MCI or as cognitively normal (CN) during standard cognitive evaluations among a large epidemiological study designed to determine prevalence and incidence of dementia and its major subtypes in Beijing, China were re-examined after an interval of approximately 3 years, repeating the same investigation protocol as at baseline. MCI subjects meeting criteria for dementia and the two major subtypes, DAT and VaD were identified at follow-up evaluation. Annual conversion rates for combined dementias and for major subtypes of DAT and VaD, from MCI, were compared with conversion rates among CN subjects. Relative risks for conversion from MCI to major subtypes of dementia were also compared with CN subjects by Cox regression models. 175 MCI and 400 CN subjects were identified at baseline. Among 121 MCI subjects available at follow-up, 51 were diagnosed with dementia (29 with DAT, 18 with VaD and 4 with other dementias), compared with 14(10 DAT, 3 VaD and 1 other type dementia) diagnosed as dementia among 281 CN subjects available at follow-up. Annual conversion rates calculated from MCI to all dementias, compared with conversion rates from CNs, were 14.1% versus 1.6%. Specifically for DAT, annual conversion rates were 8.0% versus 1.1% and for VaD were 5.0% versus 0.3%. Relative risks for developing all dementias, DAT and VaD among MCI subjects were 9, 6 and 5 times greater than among CN subjects. Conversion rates among MCI subjects to dementia, and major subtypes, for elderly Chinese residents of Beijing were comparable with results reported among similar studies worldwide. Risks of developing dementia, and major subtypes, among MCI subjects in Beijing were significantly higher than among normal subjects. Identification of MCI among elderly populations provides the possibilities for dementia prevention and treatment within prodromal stages. PMID- 16375661 TI - MRI abnormalities associated with mild cognitive impairments of vascular (VMCI) versus neurodegenerative (NMCI) types prodromal for vascular and Alzheimer's dementias. AB - BACKGROUND AND OBJECTIVES: Mild Cognitive Impairments (MCIs) are identifiable clinical entities, in neurodegenerative forms, as prodromal for Alzheimer's type (DAT) or in vascular forms, as prodromal for vascular dementia (VaD). The present longitudinal study compares and contrasts MRI abnormalities among MCI subjects as they progress to DAT versus VaD. Subjects converting to DAT and VaD confirmed ultimate diagnosis during MCI staging. In "mixed cases" the predominant MRI pathology was judged the primary cause. SUBJECTS AND METHODS: Subjects (n = 153) were selected from elderly outpatient volunteers who have been enrolled for 25 years in planned longitudinal studies of aging, stroke and dementia. Cognitively normal (CN, n = 52), MCI of neurodegenerative (N-MCI, n = 30) and vascular (V MCI), n = 35) subtypes, plus converted DAT (n = 19) and VaD (n = 17) were diagnosed according to established protocols. Combined Mini-Mental-Cognitive Capacity Screening Examinations (CMC) screened, identified and confirmed MCIs or dementias. Cerebral MRI abnormalities were analyzed utilizing volumetric measurements and visual rating scales. RESULTS: Compared with persistently cognitively normal subjects, MCI subjects and converted dementias were significantly older without significant gender differences, but cognitively impaired subjects were older than the CN group since age is a risk factor for cognitive decline. Histories of hypertension, heart disease, diabetes mellitus, TIAs and strokes were more frequent among subjects with VMCI and VaD, confirming that all vascular risk factors contribute to vascular cognitive decline, but since vascular risk factors were treated, not all progressed to VAD. Family history of neurodegenerative disease, particularly DAT, were more prevalent among NMCI and converted DAT subjects. VMCI showed more extensive leucoaraiosis and lacunar infarcts than subjects with NMCI. NMCI, prodromal for dementia of Alzheimer's type (DAT), showed more medial temporal lobe atrophy with enlarged temporal horns, and fewer vascular lesions. PMID- 16375662 TI - Mechanisms of lymphangiogenesis: targets for blocking the metastatic spread of cancer. AB - The lymphatic vasculature is an important route of metastatic spread in cancer and recent studies have demonstrated that lymphangiogenesis (the growth of lymphatic vessels) associated with tumors promotes metastasis via the lymphatics. Therefore, the molecular mechanisms that drive lymphangiogenesis are attractive targets for development of novel therapeutics designed to restrict cancer metastasis. Such therapeutics would be of high priority as metastasis is the most lethal aspect of tumor biology. Research over the past seven years has identified protein growth factors and cell surface receptors that signal for lymphangiogenesis during embryonic development, in adult tissues and in cancer. Proteases that process and thereby activate lymphangiogenic growth factors have also been defined. Lymphangiogenic growth factors, the enzymes that activate them and the cell surface receptors signalling for growth of lymphatic vessels are prime targets for anti-lymphangiogenic drugs designed to restrict cancer metastasis. Agents targeting some of these proteins have already shown promise for blocking tumor lymphangiogenesis and lymphatic metastasis in animal models. This article focuses on current and emerging targets for blocking these processes that have been defined in recent studies of the molecular mechanisms controlling lymphangiogenesis. Strategies to block the actions of these proteins in cancer are also explored. PMID- 16375663 TI - Novel therapeutic approaches targeting vascular endothelial growth factor and its receptors in haematological malignancies. AB - The existence of specific angiogenesis inhibitors was first postulated by Judah Folkman in 1971. The term "antiangiogenesis" was introduced to describe treatments designed to prevent the induction of new blood vessels and perhaps reduce the number of those already present. Several approaches inhibit tumor angiogenesis and more than 60 antiangiogenic compounds have been clinically evaluated. Because tumor-associated angiogenesis takes place in a physiological context, its inhibition should not induce resistance and should potentiate the oncostatic effect, because each neovessel supplies hundreds of tumor cells. Inhibitors may be synthetic or semi-synthetic agents, endogenous inhibitors, or biological antagonists of the angiogenic cascade. Several direct and indirect vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) inhibitor strategies are under clinical investigation for treatment of solid tumors and hematological malignancies. Approaches to disrupt the VEGF/VEGFR signalling pathways range from small-molecule ATP competitive VEGFR inhibitors to biological agents such as soluble receptors, anti-VEGF and anti-VEGFR antibodies, small molecule inhibitors, and VEGF transcription inhibitors. This review summarizes the literature on the use of these molecules in the treatment of hematological malignancies. PMID- 16375665 TI - Nuclear proteins: promising targets for cancer drugs. AB - Recent progress in cancer drug therapy has recognized that the nucleus of the eukaryotic cell is an active site for many cellular processes important to the development of cancer. Many of these processes take place in specialized compartments of the nucleus. One of such sub-nuclear compartments is the promyelocytic leukemia nuclear body (PML NB). In acute promyelocytic leukemia (APL), PML forms a fusion protein with the retinoic acid receptor (RAR) alpha as a result of chromosomal translocation. This PML-RAR alpha fusion protein is responsible for the proliferative and de-differentiated phenotype of the leukemic cells and is the target of all-trans retinoic acid (ATRA). Another example of the specialized sub-nuclear compartments important in the targeting of cancer is the nucleolus. Recently, it has been proposed that the nucleolus serves as a stress sensor for the cell, and the molecular mechanism underlying this proposal has been discovered. Moreover, many anti-cancer drugs target specific protein-protein interactions within the nucleus. We will discuss current development surrounding two such target proteins: the hypoxia-inducible factor 1 alpha (HIF-1alpha) and FKBP25. Furthermore, chromatin structure, which is affected by modifications of core histones, has become a target of anti-cancer drugs. In this review, we will emphasize the significance of nuclear proteins as promising targets for cancer drug therapy by discussing a few key ideas, in three broad categories of specialized sub-nuclear compartments, protein-protein interactions, and the modifications of the chromatin structure. PMID- 16375664 TI - Anti-angiogenic and anti-inflammatory effects of statins: relevance to anti cancer therapy. AB - Angiogenesis is indispensable for the growth of solid tumors and angiogenic factors are also involved in the progression of hematological malignancies. Targeting the formation of blood vessels is therefore regarded as a promising strategy in cancer therapy. Interestingly, besides demonstration of some beneficial effects of novel anti-angiogenic compounds, recent data on the activity of already available drugs point to their potential application in anti angiogenic therapy. Among these are the statins, the inhibitors of 3-hydroxy-3 methylglutaryl-coenzyme A reductase. Statins are very efficient in the treatment of hypercholesterolemia in cardiovascular disorders; however, their effects are pleiotropic and some are not directly related to the inhibition of cholesterol synthesis. Some reports particularly highlight the pro-angiogenic effects of statins, which are caused by low, nanomolar concentrations and are regarded as beneficial for the treatment of cardiovascular diseases. On the other hand, the anti-angiogenic activities, observed at micromolar concentrations of statins, may be of special significance for cancer therapy. Those effects are caused by the inhibition of both proliferation and migration and induction of apoptosis in endothelial cells. Moreover, the statin-mediated inhibition of vascular endothelial growth factor synthesis, the major angiogenic mediator, may contribute to the attenuation of angiogenesis. It has been suggested that the anti-cancer effect of statins can be potentially exploited for the cancer therapy. However, several clinical trials aimed at the inhibition of tumor growth by treatment with very high doses of statins did not provide conclusive data. Herein, the reasons for those outcomes are discussed and the rationale for further studies is presented. PMID- 16375666 TI - Therapeutic cancer vaccines: at midway between immunology and pharmacology. AB - The pursuit of active specific immunotherapy of cancer re-emerged vigorously in the 90s. More than 50 vaccines are currently under clinical testing, and more than 400 clinical trials have been conducted. This wave of enthusiasm is rooted in fundamental immunology, as new paradigms, such as the dominant tolerance through T-regulatory cells and the instructive role of the innate immune system on the adaptive immune system, opened the possibility that an efficient cancer vaccination could be achieved even without the need of cancer neoantigens, provided that antigen presentation could be increased, and that regulatory circuits could be controlled. However, recent failures in some large trials have brought disappointment and have highlighted the differences between experiments in young, healthy mice with small transplanted tumours, and clinical testing in aged, ill patients with advanced spontaneous tumours, driving the attention to issues such as tumour editing, tumour-induced immunosuppression, and immunosenescence. The molecular basis of these phenomena is only partially known. Additionally, the inherent complexity of the immune system as a network of multiple interactions and redundant control loops among a huge diversity of components sets another barrier to the translation of in vitro reductionist knowledge into rationally designed clinical trials. All this calls for a new therapeutic paradigm in cancer vaccines, moving beyond the analogy with the classic drug-target approach, and targeting the immune system regulation as a whole, and its interaction with the tumour, in all its complexity. Early mathematical modelling of cancer immunotherapy has suggested how to go about it. This re-evaluation of the cancer vaccine landscape, suggests that future successful cancer immunotherapy will be combined immunotherapy, will be exquisitely schedule-dependent and will need new experimental models allowing for the exploration of the mechanisms of resistance and tumour escape, such as tumour editing and tumour induced immunosuppression, in the context of the physiology of the immune system of the elderly. These shifts will put cancer vaccines closer to pharmacology than to conventional preventive vaccinology, or at least at the midway. A change in the design and the ultimate goals of the clinical trials will also be needed, identifying long term stabilization of the disease and quality of life as main endpoints, again closer to the clinical management of most chronic noncommunicable diseases. PMID- 16375667 TI - Mechanisms of focal adhesion kinase regulation. AB - Focal adhesion kinase (FAK) is a tyrosine kinase whose phosphorylation state and activity is tightly linked to cell adhesion to the extracellular matrix through integrin receptors. FAK's regulation by adhesion places it in a key position to be able to influence cellular events that are either dependent on cell adhesion like cell proliferation and survival, or that require modulation of cell adhesion like cell migration. FAK's involvement in cellular pathways that regulate cell growth and cell movement suggests that it may contribute to the development of cancer or other diseases. FAK's possible involvement in these pathways makes it a potential drug target. In this review we will focus on the developing view how FAK's activity and phosphorylation are regulated within the cell. Specifically, we will address the contribution of integrins and growth factor dependent pathways to FAK's activation. The role of the tyrosine kinase Src in FAK's regulation will be discussed. The contribution of various negative regulators of FAK's phosphorylation on its regulation including phosphatases and proteases will be discussed. Lastly, the emerging role of FAK's amino terminal FERM like domain in FAK's regulation will be explored. FAK's function within a cell are tightly linked to its phosphorylation state, thus understanding its normal regulation in the cell will provide important insight into drug development by highlighting novel regulatory mechanisms within FAK that potentially may be exploited. PMID- 16375668 TI - Fungal infections and antifungal strategies. PMID- 16375669 TI - Fungal infections of the skin: infection process and antimycotic therapy. AB - Dermatomycoses are among the most widespread and common superficial and cutaneous fungal infections in humans. These typically nonfatal conditions are difficult to treat, especially infections of the nail. Dermatomycoses are caused by filamentous fungi such as Trichophyton, Microsporum or Epidermophyton species. These filamentous fungi have a high affinity for keratin, an important component of hair, skin and nails, which are the primary areas of infection by dermatophytes. The antifungal agents currently marketed for dermatomycoses are mainly inhibitors of ergosterol biosynthesis, except for griseofulvin, which interferes with the cytoplasmic and nuclear microtubular system. Three different types of inhibitors of the ergosterol biosynthetic pathway have been proven to be effective in clinic: the azoles (e.g. topical miconazole and topical/oral ketoconazole, itraconazole and fluconazole), the allylamines (e.g. terbinafine) and morpholines (amorolfine). Even today more effective antifungal azoles with less adverse effects and short-term therapy are deemed necessary to treat dermatophytosis. A promising novel triazole compound in this respect is R126638, which showed potent in vitro and in vivo activity. PMID- 16375670 TI - Systemic fungal infections caused by Candida species: epidemiology, infection process and virulence attributes. AB - Candida species, in particular C. albicans, represent a major threat to immunocompromised patients. Able to exist as a commensal on mucosal surfaces of healthy individuals, these opportunistic fungi frequently cause superficial infections of mucosae and skin. Furthermore, in hospital settings, Candida species may cause life-threatening invasive infections in a growing population of vulnerable patients. In fact, candidaemia is associated with the highest crude mortality of all bloodstream infections. Candida cells may enter the bloodstream by direct penetration from epithelial tissues, due to damage of barriers in the body caused by surgery, polytrauma or drug treatment, or may spread from biofilms produced on medical devices. From the bloodstream, cells may infect almost all organs but appear to prefer certain organs depending upon the route of infection. The exact mechanisms by which Candida cells survive the challenge of the blood environment and escape from the bloodstream to cause deep-seated infections have not yet been elucidated, but various investigations are reviewed. It is clear, however, that Candida must have particular attributes which enable the organism to survive and grow within the environment of healthy individuals and to invade tissues in the immunocompromised host. Most studies have focussed on C. albicans and this review will therefore summarise work on the various known virulence factors and methods used to identify further virulence attributes of this fungus. PMID- 16375671 TI - Systemic fungal infections caused by Aspergillus species: epidemiology, infection process and virulence determinants. AB - Infections with mould pathogens have emerged as an increasing risk faced by patients under sustained immunosuppression. Species of the Aspergillus family account for most of these infections and in particular Aspergillus fumigatus can be regarded as the most important airborne-pathogenic fungus. The improvement in transplant medicine and the therapy of hematological malignancies is often complicated by the threat of invasive aspergillosis. Specific diagnostics are still limited, as are the possibilities of therapeutic intervention. Hence, invasive aspergillosis is still associated with a high mortality rate that ranges from 30 % to 90 %. In recent years, considerable progress has been made in understanding the genetics of A. fumigatus and molecular techniques for the manipulation of the fungus have been developed. Molecular genetics offers not only approaches for the detailed characterization of gene products that appear to be key components of the infection process but also selection strategies that combine classical genetics and molecular biology to identify virulence determinants of A. fumigatus. The review discusses aspects of the current knowledge of the infection process, mechanisms of protection of the fungus against immune effector cells, and virulence determinants of A. fumigatus. PMID- 16375672 TI - Fungal biofilms and antimycotics. AB - Device-related infections in most nosocomial diseases can be traced to the formation of biofilms (microbial communities encased within a polysaccharide-rich extracellular matrix) by pathogens on surfaces of these devices. Candida species are the most common causative agents of these infections, and biofilms formed by these fungal organisms are associated with drastically enhanced resistance against most antimicrobial agents. This enhanced resistance contributes to the persistence of this fungus despite antifungal therapy. Recent studies showed that Candida biofilms exhibit antifungal resistance against most antifungal agents with the exception of echinocandins and lipid formulations of AMB. This review discusses methods used to evaluate biofilm resistance and provide information on susceptibility pattern of candidal biofilm as well as studies investigating the mechanisms underlying biofilm resistance. PMID- 16375673 TI - Currently used antimycotics: spectrum, mode of action and resistance occurrence. AB - The increasing incidence of fungal infections combined with the emerging problem of antifungal drug resistance have prompted investigations of the mode of action of the currently used antifungal therapeutics (antimycotics). The routinely used antimycotics can be grouped into six different classes based on their mode of action. In this review, the mode of action and antifungal spectrum of these classes are discussed, together with possible resistance development against them. PMID- 16375674 TI - Fungal pathogens research: novel and improved molecular approaches for the discovery of antifungal drug targets. AB - With the rise of fungal infection incidence amongst the patient population, the importance of developing new antifungal drug targets is higher than ever. This review mainly focuses on the three most prevalent fungal pathogens, Candida, Aspergillus and Cryptococcus, and on the most recent progresses in molecular research that contribute to a better understanding of the pathogen itself, but also its host and the interaction with its host. We consider the progress made in comparative genomics following the huge effort of fungal genome sequence projects undertaken in the last few years. We focus not only on currently used mammalian animal models such as mice, but also on novel non-mammalian models, such as the nematode worm Caenorhabditis elegans and the fruit fly Drosophila melanogaster, which offer useful tools in the area of the innate immune response to fungal infections. In addition we relate to the recent genomic and proteomic studies and focus on the use of these approaches in in vivo experiments in the pathogen itself as well as in the host. Finally, we describe the latest targeted mutagenesis strategy available in C. albicans and the use of RNA interference in both Cryptococcus neoformans and A. fumigatus. Our aim is not to give an exhaustive list of all new strategies but rather to give an overview of what will contribute most to the identification of new antifungal drug targets and the establishment of novel antifungal strategies. PMID- 16375675 TI - Fungal sphingolipids as targets for the development of selective antifungal therapeutics. AB - Sphingolipids are essential membrane components, present in all eukaryotic cells, but structurally distinct in mammalian and fungal cells. Therefore, they represent an attractive new target for the development of novel antimycotics. This review will briefly highlight sphingolipid biosynthesis and functions in the yeast Saccharomyces cerevisiae. In addition, naturally occurring antifungal compounds that interact with fungal-specific sphingolipids, resulting in fungal growth arrest, will be discussed regarding their mode of action, and therapeutic value. These compounds include plant and insect defensins, syringomycin E and antifungal antibodies to sphingolipids. PMID- 16375676 TI - Antifungal susceptibility testing methods. AB - The number of systemically active antifungal agents has increased dramatically in recent years in response to the challenge of invasive mycoses. Additional work is needed to better understand the mechanisms of action of these agents as well as the mechanisms of resistance expressed by the fungal pathogens. Antifungal susceptibility testing has been standardized and refined and now may be considered to play an important role in the management of invasive mycoses. More work is needed to optimize the methods for testing new antifungal agents and for testing pathogens other than Candida. The ongoing efforts and international collaborations designed to address these issues will provide important information that will improve the management of serious fungal infections. PMID- 16375677 TI - Detection of fungal infections using radiolabeled antifungal agents. AB - The outcome of antifungal therapy depends on the progression of the infection at the start of therapy. Unfortunately, most patients are diagnosed once the fungal infection has progressed considerably as a result of the non-specific clinical signs of fungal infections in immunocompromised patients and the poor sensitivity of current mycological diagnostic tests. This review will highlight current fungal diagnostic techniques and will focus on scintigraphic methods for the specific detection of fungal infections in mice. For this purpose, antifungal components (e.g. fluconazole and antifungal peptides) are radiolabeled e.g. with technetium-99m ((99m)Tc) and their in vivo distribution is monitored in infected mice. It has been demonstrated that (99m)Tc-fluconazole is an excellent tracer to detect Candida albicans infections in mice as it distinguishes these infections from bacterial infections and sterile inflammations. However, this radiopharmaceutical only poorly detects infections with Aspergillus fumigatus in mice. (99m)Tc-peptides derived from antifungal peptides/proteins, such as human ubiquicidin and lactoferrin, can distinguish C. albicans and A. fumigatus infections from sterile inflammations, but not from bacterial infections, in mice. Furthermore, the efficacy of fluconazole in C. albicans-infected mice could be successfully monitored using (99m)Tc-ubiquicidin. In conclusion, neither (99m)Tc-fluconazole nor the (99m)Tc-peptides tested are optimal tracers for fungal infections. Nonetheless, since early initiation of antifungal therapy for candidemia reduces its high mortality rate, a positive result with (99m)Tc fluconazole scintigraphy is of clinical relevance. Finally, the possibility that other (radiolabeled) antifungal agents, e.g. voriconazole, caspofungin, antifungal plant or insect defensins, can be useful for detection of fungal infections should be considered. PMID- 16375678 TI - The endocannabinoid system in the brain: from biology to therapy. PMID- 16375679 TI - Endocannabinoid metabolic pathways and enzymes. AB - Endocannabinoids, defined in 1995 as endogenous agonists of cannabinoid receptors, their anabolic and catabolic pathways, and the enzymes involved in these pathways (the "endocannabinoid enzymes"), are the subject of this review. A general strategy seems to apply to the regulation of the levels of the two major endocannabinoids, anandamide and 2-arachidonoylglycerol (2-AG). Five endocannabinoid enzymes have been cloned to date: two are responsible for the biosynthesis and degradation of anandamide, the NAPE-selective phospholipase D and the fatty acid amide hydrolase, respectively; the other three catalyse the biosynthesis and degradation of 2-AG, the sn-1-selective diacylglycerol lipases alpha and beta and the monoacylglycerol lipase, respectively. The major features of these five proteins, their relative weight in determining endocannabinoid levels, and the possible targeting of some of them for therapeutic purpose, as well as the possibility of the existence of alternative anabolic and catabolic pathways are discussed. PMID- 16375680 TI - Molecular biology of the enzymes that degrade endocannabinoids. AB - Endocannabinoid circuits have been shown to regulate a number of important pathways including pain, feeding, memory and motor coordination. Direct manipulation of endocannabinoid tone, therefore, may relieve disease symptoms related to analgesia, obesity, Alzheimer's and Parkinson's in humans. The endocannabinoid circuit involves two cloned receptors: CB1 in the CNS and CB2 in the periphery; endogenously produced ligands including anandamide, 2 arachidonylglycerol and palmitoylethanolamide; and enzymes that degrade endocannabinoid ligands to terminate signaling. Currently, three enzymes have been characterized with the ability to hydrolyze endocannabinoids: fatty acid amide hydrolase (FAAH), monoglyceride lipase (MGL) and N-acylethanolamine hydrolyzing acid amidase (NAAA). The purpose of this review is to examine the molecular biology for the enzymes that hydrolyze endocannabinoids covering the protein activity and expression, mRNA characterization, genomic locus organization, promoter analysis and knockout phenotypes. PMID- 16375681 TI - Endocannabinoids in the central nervous system: from neuronal networks to behavior. AB - Retrograde synaptic signaling influences both short-term and long-term plasticity of the brain, in both excitatory and inhibitory synapses. During the last few years it has become apparent that the endogenous ligands for the cannabinoid CB1 receptor, the "endocannabinoids", fulfill an essential role in the brain as retrograde synaptic messengers, in a number of structures including the hippocampus, cerebellum and the limbic and mesocortical systems. This seminal discovery provides a cellular basis for the well known ubiquitous role of the endocannabinoids and their receptors (together, the "ECBR" system) in virtually all brain functions studied. This review will relate the anatomical distribution of the endocannabinoids and their CB1 receptors to functions of the ECBR system, as much as possible in light of the endocannabinoids as retrograde synaptic messengers. Functional implications of the high rates of co-localization with cholecystokinin (CCK), will also be considered. The most obvious function to be profoundly affected by the retrograde synaptic role of the endocannabinoids is memory. However, additional functions and dysfunctions such as reward and addiction, motor coordination, pain perception, feeding and appetite, coping with stress, schizophrenia and epilepsy will also be reviewed. Finally, the widespread presence of the ECBR system in the brain also lends a scientific basis for the development of cannabinoid-based medicines. The same ubiquity of the ECBR system however, should also be taken into consideration with respect to possible adverse side effects and addictive potential of such pharmaceutical developments. PMID- 16375682 TI - Endocannabinoids and drug dependence. AB - Drug dependence is a chronically relapsing disorder, manifested as an intense desire for the drug, with impaired ability to control the urges to take the drug, even at the expense of serious adverse consequences. These behavioral abnormalities develop gradually during repeated exposure to a drug of abuse, and can persist for months or years after discontinuation of use, suggesting that this addiction can be considered a form of drug-induced neural plasticity. Many neurotransmitters, including gamma-aminobutyric acid (GABA), glutamate, acetylcholine, dopamine, serotonin and endogenous opioid peptides, have been implicated in the effects of the various drugs of abuse. Dopamine has been consistently associated with the reinforcing effects of most of them. There is, in addition, a growing body of evidence that the endogenous cannabinoid system might participate in the motivational and dopamine-releasing effects of several drugs of abuse. This review will discuss the latest advances on the mechanisms of cannabinoid dependence and the possible role of the endocannabinoid system in the treatment of addiction, not only to marijuana but also to the other common illicit drugs. PMID- 16375683 TI - Endocannabinoids in neuroimmunology and stress. AB - Two topics are presented in this review. In the first section, we review data regarding the effects of the endocannabinoids (eCBs) and cannabinoid receptors on neuroimmune function. The function of eCBs in the interaction between the immune system and the central nervous system (CNS) is of particular interest, since the CNS itself is a rich source of eCBs while being exquisitely sensitive to inflammation. There are several sites at which cannabinoids can influence neuroinflammation. Microglial cells express both CB receptors and make eCBs. Activation of CB receptors on these cells seems to promote migration and proliferation but to reduce activation to macrophages. In several neurodegenerative diseases, up-regulation of microglial CB2 receptors have been observed. It is our hypothesis that microglial CB receptor activity is anti inflammatory and could be exploited to manipulate neuroinflammatory processes with a minimum of unwanted effects. The second topic discussed suggests that the eCB/CB1 receptor pair is involved in the responses of animals to acute, repeated and variable stress. The roles of this pair are complex and dependent upon previous stress, among other things. Dysfunctional responding to stress is a component of several human neuropsychiatric disorders, including anxiety and panic disorders, post-traumatic stress disorders, premenstrual dysphoria and quite possibly, drug abuse. While it is too early to say with certainty, it is very possible that either inhibition or potentiation of endocannabinoid signaling will be an efficacious novel therapeutic approach to more than one human psychiatric disease. PMID- 16375684 TI - From cannabis to endocannabinoids in multiple sclerosis: a paradigm of central nervous system autoimmune diseases. AB - An increasing body of evidence suggests that cannabinoids have beneficial effects on the symptoms of multiple sclerosis, including spasticity and pain. Endogenous molecules with cannabinoid-like activity, such as the "endocannabinoids", have been shown to mimic the anti-inflammatory properties of cannabinoids through the cannabinoid receptors. Several studies suggest that cannabinoids and endocannabinoids may have a key role in the pathogenesis and therapy of multiple sclerosis. Indeed, they can down regulate the production of pathogenic T helper 1 associated cytokines enhancing the production of T helper 2-associated protective cytokines. A shift towards T helper 2 has been associated with therapeutic benefit in multiple sclerosis. In addition, cannabinoids exert a neuromodulatory effect on neurotransmitters and hormones involved in the neurodegenerative phase of the disease. In vivo studies using mice with experimental allergic encephalomyelitis, an animal model of multiple sclerosis, suggest that the increase of the circulating levels of endocannabinoids might have a therapeutic effect, and that agonists of endocannabinoids with low psychoactive effects could open new strategies for the treatment of multiple sclerosis. PMID- 16375685 TI - Cannabinoids: between neuroprotection and neurotoxicity. AB - Cannabinoids, such as the delta9-tetrahydrocannabinol (THC), present in the cannabis plant, as well as anandamide and 2-arachidonoyl glycerol, produced by the mammalian body, have been shown to protect the brain from various insults and to improve several neurodegenerative diseases. The current review summarizes the evidence for cannabinoid neuroprotection in vivo, and refers to recent in vitro studies, which help elucidate possible molecular mechanisms underlying this protective effect. Some of these mechanisms involve the activation of CB1 and CB2 cannabinoid receptors, while others are not dependent on them. In some cases, protection is due to a direct effect of the cannabinoids on neuronal cells, while in others, it results from their effects on non-neuronal elements within the brain. In many experimental set-ups, cannabinoid neurotoxicity, particularly by THC, resides side by side with neuroprotection. The current review attempts to shed light on this dual activity, and to dissociate between the two contradictory effects. PMID- 16375686 TI - Pharmacological properties and therapeutic possibilities for drugs acting upon endocannabinoid receptors. AB - Clinical trial data are beginning to emerge with respect to the therapeutic efficacy of cannabis extracts for the treatment of chronic pain. Although there is some evidence of efficacy, a major issue concerns the narrow margin between doses producing therapeutic effects and those producing the "highs" associated with cannabis misuse. In addition, long-term use is associated with an increased risk of psychiatric illness. These negative aspects constrain the doses of cannabis extracts and psychoactive cannabinoids that can be given to patients, and raise the risk that properly conducted clinical trials with too low dosages will impact negatively on subsequent drug development in this field. However, recent research has opened up a number of avenues whereby compounds acting directly upon cannabinoid (CB) receptors may have therapeutic potential. In this review, two such areas are discussed, namely a) the possible use of peripherally acting CB agonists and CB2 receptor-selective agonists for the treatment of pain, and b) the possible utility of CB2 receptor agonists for the prevention of stress induced exacerbations of skin disorders such as psoriasis. A second area of drug development at present is that of CB1 receptor antagonists/inverse agonists, spearheaded by rimonabant, for the treatment of obesity and as an aid for smoking cessation. An important aspect of these compounds is their efficacy and selectivity, and this is discussed in detail in the present review. PMID- 16375687 TI - Therapeutic perspectives of inhibitors of endocannabinoid degradation. AB - The study of the cannabinoids can be established in the middle sixties with the elucidation of the structure of the active principle of Cannabis sativa plant, the delta9-tetrahydrocannabinol. However, the existence of an endogenous cannabinoid system (ECS) has not been unequivocally accepted until recently. The last two decades have witnessed an impressive advance in the knowledge about cannabinoids, their chemistry, the enzymes involved in their metabolism, and their physiological and pathological roles. In particular, we have made progress in modifying the activity of the ECS with selective compounds, validating the ECS as a new therapeutic target. Endocannabinoids play a role in physiological and pathological processes, and their levels are affected in several disorders. Therefore, it should be possible to ameliorate these pathologies by correcting their altered levels. This review focuses on the current therapeutic opportunities of endocannabinoid-directed drugs, and pays special attention to the therapeutic possibilities underlying the inhibition of the endocannabinoid inactivation. The strategy of manipulating the ECS might open new avenues in the development of therapeutic approaches for a number of disorders, both central and peripheral, that lack as yet effective treatments. PMID- 16375688 TI - Partial QSAR analysis of some selected natural inhibitors of FAAH suggests a working hypothesis for the development of endocannabinoid-based drugs. AB - The endogenous cannabinoids (endocannabinoids) are bioactive signaling molecules, that show diverse cellular and physiological effects and play various roles in the central nervous system, as well as in the periphery. The discovery of N arachidonoylethanolamine (anandamide, AEA) and of the enzyme that terminates its signaling, i. e. fatty acid amide hydrolase (FAAH), has inspired pharmacological strategies to augment endocannabinoid tone and biological activity through inhibition of FAAH. Here we discuss the role of natural endocannabinoid derivatives, like the hydroxy-anandamides (OH-AEAs) generated from AEA via lipoxygenase activity, as powerful inhibitors of FAAH. We propose that these compounds, by reversibly inhibiting FAAH, may control in vivo the endocannabinoid tone. We consider the theoretical structural properties of OH-AEAs and other natural inhibitors of FAAH, based on the calculation of theoretical molecular descriptors commonly used in Quantitative Structure Activity Relationship (QSAR) studies. The QSAR properties of OH-AEAs and congeners suggest that they could act at different specific sites of FAAH, thus confirming their potential value as templates for the development of next-generation therapeutics. PMID- 16375689 TI - Amino Acid Transporter ATB0,+ as a delivery system for drugs and prodrugs. AB - ATB(0,+) is a unique amino acid transporter because of its broad substrate specificity and concentrative ability. This transporter recognizes neutral as well as cationic amino acids. It is energized by Na(+) and Cl(-) gradients and membrane potential. Many of the amino acids and amino acid derivatives that are substrates for ATB(0,+) serve as therapeutic agents (e.g., D-serine, carnitine, and nitric oxide synthase inhibitors). Recent studies have shown that the potential of ATB(0,+) as a drug delivery system may be greater than previously envisaged. ATB(0,+) can transport antiviral drugs such as acyclovir and ganciclovir when they are covalently coupled to the side chain of anionic amino acids. Chemical modification of the carboxyl groups in the side chain of aspartate and glutamate with drugs converts these anionic amino acids into neutral amino acid derivatives. Therefore, the modified drugs are recognized by ATB(0,+). Interestingly, even when acyclovir and ganciclovir are coupled as esters with alpha-carboxyl group of neutral amino acids, the modified drugs are transported via ATB(0,+). Similarly, the hydroxyl group in the side chains of serine and threonine can also be used to covalently couple drugs for delivery into cells via ATB(0,+). This increases the potential for designing a wide variety of amino acid-based prodrugs that can utilize ATB(0,+) as drug delivery system. Furthermore, the transporter is expressed in the colon, lung, and eye, the tissues easily amenable for drug delivery. These findings argue strongly in support of ATB(0,+) as a potential delivery system for a wide variety of drugs and prodrugs. PMID- 16375690 TI - Augmenting T helper cell immunity in cancer. AB - Cancer specific immunity elicited with vaccines has traditionally focused on the activation of the CD8 cytolytic T lymphocyte (CTL) often involving direct stimulation of immunity using HLA-class I binding peptide epitopes. Recently it has become clear that activation of the CTL immune effector arm alone is insufficient to mediate an anticancer response. A major problem is that CD8 T cells alone can not be sustained without the concomitant activation of CD4 T helper (Th) cells. In fact, it is now widely recognized that the Th cell regulates nearly all aspects of the adaptive immune response. In addition, Th cells can recruit the innate immune system during immune augmentation. Therefore, the focus of the immune response in cancer has shifted away from activating CTL immunity alone to activating Th cell immunity alone or concurrently with CTL. Evidence suggests that activating the Th cell is sufficient to get a complete adaptive immune response because, once activated, the Th cell will elicit endogenous CD8 T cell and humoral immunity. In this review, we discuss the role of the Th cell in the adaptive immune response to cancer, how peptides that are capable of activation of Th cells are identified, and the clinical translation of newly identified candidate Th cell peptide epitopes to human cancer specific vaccines. Over the next decade, studies should begin to further define how we can manipulate the Th immune effector arm to achieve effective antitumor immunity. PMID- 16375691 TI - Management of liver disease in thalassemia: main drug targets for a correct therapy. AB - Liver disease is the second cause of mortality in thalassemia major. We present a review on the hepatic damage in thalassemic patients aimed at a knowledge of current preventive, diagnostic and therapeutic approaches, useful to guide in clinical judgment and treatment decisions. Transfusion related iron overload and hepatitis are the causes of liver damage in thalassemic patients. We examined means of primary prevention, anti-hepatitis vaccinations, blood donors screening; diagnostic tests for secondary prevention (computed tomography, magnetic resonance imaging, super conducting quantum interference device and biopsy) were also discussed about. A survey of treatment methods and strategies ( chelation therapy, antiviral treatments and liver and bone marrow transplantation) follows. PMID- 16375693 TI - The new immunosuppression: intervention at the dendritic cell-T-cell interface. AB - Dendritic cells (DCs) play an important role in initiating and directing T-cells towards immunity or tolerance. An important aim of emerging immunosuppressive strategies is to ensure that antigen is perceived in a 'tolerogenic context'. This would have obvious benefit in minimising the need for long-term drug maintenance in organ transplantation, hypersensitivity and autoimmune diseases. Here we review the biology of the interplay between the DC and T-cell, with a specific focus on therapeutic drugs targeting molecules that effect their interaction and function. PMID- 16375692 TI - Molecular basis of therapeutic strategies for breast cancer. AB - The development of breast cancer is the consequence of uncontrolled growth and division of breast-ductal epithelial cells. While many factors contribute to its etiology, estrogen hormones within the context of many interrelated growth signaling pathways play critical roles for the initiation and development of breast cancer. The effects of estrogens are primarily mediated by the estrogen receptors (ERs) alpha and beta. ER mediates a complex array of genomic and non genomic events that orchestrate cellular metabolism, mitogenesis, morphogenesis, motogenesis, and apoptosis. The current modalities for the treatment of breast cancer have centered on the development of agents with diverse pharmacology to reduce/ablate the circulating estrogens or to alter/prevent ER function. Approaches to perturb the estrogen environment are successful usually in the remission of established tumors. However, many breast tumors are not responsive or eventually develop resistance to endocrine therapies. Despite considerable effort, the mechanism for the non-responsiveness and acquisition of resistance remains unclear. The establishment of hormone responsiveness is one of the current approaches for the development of an effective therapeutic modality for de novo resistant breast tumors. Re-establishment of loss of ER synthesis/function, on the other hand, constitutes a primary therapeutic goal for acquired resistance neoplasms. We have recently engineered transregulatory proteins that specifically targeted and robustly regulated estrogen responsive genes independent of ligand, ER-subtype and cell-context. The targeted regulation of estrogen responsive gene networks by these designer transregulators could provide a basis for the development of novel approaches for experimental biology and medicine. PMID- 16375694 TI - Failure of immune homeostasis -- the consequences of under and over reactivity. AB - The immune system is a tightly regulated network that is able to maintain a balance of immune homeostasis under normal physiological conditions. Normally, when challenged with foreign antigen, specific appropriate responses are initiated that are aimed at restoring homeostasis. However under particular circumstances, this balance is not maintained and immune responses either under or over react. Cancer is an example of a situation where the immune response can be inefficient or unresponsive, resulting in uncontrolled growth of the cancer cells. Conversely, when the immune response over-reacts, this can result in conditions such as autoimmunity or pathology following infection. Many drug therapies have been developed that aim to alleviate or prevent such immune disorders and restore immune homeostasis. This review highlights recent advances in immunotherapies, with an emphasis on specific examples in the treatment of cancer, autoimmune disease (multiple sclerosis) and viral infection (respiratory syncytial virus). PMID- 16375695 TI - Insulin regulation of PEPCK gene expression: a model for rapid and reversible modulation. AB - Insulin and glucagon regulate the expression and/or activity of a variety of proteins to maintain blood glucose within normal limits. A key target is the gene encoding phosphoenolpyruvate carboxykinase (PEPCK), which catalyzes the first committed step in hepatic gluconeogenesis. Acute regulation of PEPCK is achieved by modulating transcription of the gene, which is tightly regulated by cAMP (the mediator of glucagon and catecholamines), glucocorticoids and insulin. Normally, PEPCK expression is induced by glucagon, catecholamines and glucocorticoids during periods of fasting and in response to stress, but is dominantly inhibited by glucose-induced increases in insulin secretion upon feeding. The incomplete effectiveness of insulin action, whether due to intermittent insulin injection in type I diabetics or insulin resistance in type II diabetics, contributes to hyperglycemia and complications, resulting in damage to the eyes, nerves, kidneys and other organs over time. Thus, defining a molecular mechanism for insulin inhibition of PEPCK gene transcription has been a major goal of research in several labs, because it would allow the development of drugs to prevent episodic increases in circulating glucose in diabetics. Here, we review the main lines of investigation into this complex problem and the likely properties of an inhibitor. Any mechanism must account for the rapidity, specificity and dominance with which insulin is known to act in regulating PEPCK transcription. To date Foxo1 (FKHR) is the only transcription factor for which a complete path from the insulin receptor to gene regulation has been described. While this explains the regulation of some genes, such as IGFBP-1, Foxo1 appears not to play a requisite role in regulating PEPCK transcription. Investigation of cis-acting elements in the PEPCK promoter has shed considerable light on the mechanisms of activation by cAMP and glucocorticoids but has failed to identify a regulatory element that mediates insulin inhibition of transcription. This, together with evidence from analysis of the inducing mechanisms, has prompted us, and others, to investigate the possibility that insulin disrupts activation rather than independently promoting repression. Thus, we hypothesize that insulin-induced modification of a key transcription regulatory protein prevents an essential factor from participating in the induction process, leading to rapid but reversible inhibition, as is seen in animals. The ability to alter the sensitivity of a key transcription factor to improve insulin-regulated control of blood glucose would be a major improvement in the treatment of diabetes, a growing problem in the industrialized world. PMID- 16375696 TI - Hormonal effects on drug metabolism through the CYP system: perspectives on their potential significance in the era of pharmacogenomics. AB - Cytochrome P450 (CYP) is a group of enzymes that metabolize drugs to a more water soluble form, rendering them available for renal excretion. The major site of CYP expression is the liver. Nearly 50% of all medications currently on the market are metabolized by the enzyme CYP3A4, while metabolism of another 35-40% occurs through enzymes CYP1A2, CYP2C19, CYP2D6, CYP3A5 CYP3A6, and CYP3A7. Here, we summarize the current knowledge of the effects of hormones on the CYP family. The term "hormone" is used in its broad sense and includes products of the major endocrine glands (i.e., thyroid, adrenals, gonads, pancreas) and compounds that are not classically considered hormones, such as neurogenic amines, cytokines, interleukins, and eicosanoids. In addition, we comment on the effects on CYP expression of states associated with profound hormonal changes, such as pregnancy, malnutrition, obesity, diabetes mellitus, systemic inflammation, and conditions of altered extracellular fluid volume or osmolality. Available data are limited and are derived primarily from in vitro and animal studies. Moreover, the picture is obscured by conflicting results among studies and the complexity of the regulation of the expression and activity of elements of the CYP system. While the clinical significance of hormonal effects on the CYP system remains to be determined, we anticipate that such effects will be most pertinent to drugs with a narrow therapeutic range. Further research is needed to determine the scope and significance of these effects in view of rapid advances in the field of pharmacogenomics and the ever-increasing number of drugs available for therapeutic use. PMID- 16375697 TI - Signal transducer and activator of transcription 5 (STAT5), a crucial regulator of immune and cancer cells. AB - STAT5 belongs to a small family of transcription factors with dual functions. The seven signal transducers and activators of transcription (STAT) act as signaling components between the plasma membrane and the nucleus, and as transcription factors with specific DNA binding ability in the nucleus. STAT5 regulates the expression of genes, which determine important cellular phenotypes. It can promote proliferation and inhibit apoptosis, but is also involved in the regulation of differentiation between specific gene expression. STAT5 can also contribute to the transformed phenotype. In many leukemias and some solid tumors, STAT5 is constitutively activated through receptors or receptor associated tyrosine kinases and contributes to the survival and the proliferation of malignant cells. STAT5 activity appears to be limiting for these phenotypes. Inhibition of STAT5 in these tumor cells results in growth arrest and apoptosis. Targeting of STATs and other downstream mediators of oncogenic tyrosine kinases provides a promising strategy for tumor therapy, which might be refractory to resistance mechanisms incapacitating tyrosine kinase inhibitors. The well-studied steps in the activation of STAT5 and its roles in different subcellular compartments suggest original interference strategies, which could be used to inhibit its function. The challenge for drug developers will be the exploitation of defined protein-protein or protein-DNA interactions as targets of inhibition. PMID- 16375698 TI - Boosting interleukin-10 production: therapeutic effects and mechanisms. AB - More than forty cytokines have been extensively researched on the molecular structure, cell signaling and transduction pathway. With respect to cytokine regulating therapy in immunological imbalance however, the reported results are conflicting because of the pleiotropic functions and the intricate interactions of the cytokine network. In this review, we outline the observations on interleukin-10 (IL-10) upregulatory therapy. Despite varying opinions on its therapeutic effects for different disorders, IL-10 has been considered a potential anti-inflammatory cytokine. Numerous studies support the view that IL 10 shows a strong suppressive effect on Th1 lymphocytes, antigen presenting cells and the production of inflammatory mediators. It is also noticeable that recent research has revealed the relationship between IL-10 induced antigen specific regulatory CD4+ T cells and antigen specific immune tolerance. This specific regulation was mediated in part through IL-10 secretion, because anti-IL-10 treatment reverted the inhibitory effect of regulatory T cell clones. In different models, these cells were shown to inhibit both Th1 and Th2-type inflammatory responses through the secretion of IL-10. With the presence of IL 10, regulatory T cells may induce peripheral immune tolerance. Exogenous administration, transgenic expression and endogenous stimulative agents of IL-10 have been used for a variety of inflammatory diseases, autoimmune diseases and allograft rejection in patients and experimental models. A therapeutic intervention with drug inducing endogenous IL-10 may be more practical than an exogenous administration of IL-10 with transient effect. Although further investigation on gene regulation of IL-10 is necessary, increasing studies have been reported concerning the attempt to develop the agents, which could promote endogenous IL-10 production for the treatment of immunological disorders and inflammatory diseases. With some unclear mechanisms, these agents have strongly upregulated IL-10 production in vitro or in vivo. Reported IL-10 upregulatory agents have shown promising prospects for remission of autoimmune diseases and inflammatory diseases and have even induced antigen specific immune tolerance. It is interesting that the IL-10 upregulatory effect of several traditional immunosuppressive drugs has been detected, e.g. glucocorticoid, which is considered "not more as an immunosuppressive drug but an immune modulating agent". Approximately twenty IL-10 upregulatory agents as instances are described in the present review. In addition, their therapeutic effects in various diseases are discussed. PMID- 16375699 TI - Development of lymphatic vessels: tumour lymphangiogenesis and lymphatic invasion. AB - In human solid cancer, the lymph node status is the most important prognostic indicator for the clinical outcome of patients. Follow-up data has shown that about 80% of metastasis follows an orderly pattern of progression via the lymphatic network while about 20% systemic metastasis occurs, bypassing the lymphatic system. Over the past few years, advances have been made in understanding the cellular and molecular aspects of physiological lymphangiogenesis and tumour-induced lymphangiogenesis, and the majority of studies point out to a positive correlation between tumour-induced lymphangiogenesis and lymphatic metastasis. However, the impact of intra- and peritumoural lymphatics on the tumour biology and the first steps of lymphatic metastasis, i.e. the invasion of tumour cells into the lymphatic vessels, are not well understood. We will give an outline of i. the physiological process of lymphangiogenesis, ii. tumour-induced lymphangiogenesis and lymphatic metastasis, iii. lymphatic invasion and the common pathways of tumour-lymphangiogenesis and lymphatic invasion. The growing interest in this topic has brought up a number of new molecular players in the field, which may provide the basis for a rational therapy against the process of lymphatic dissemination of tumour cells. PMID- 16375700 TI - Neuropeptide-derived antimicrobial peptides from invertebrates for biomedical applications. AB - Since the beginning of the 20th century, important medicinal progress has led medical doctors to think that the end of devastating epidemics has arrived. In 1930, the discovery of sulfamides and penicillin opened a wide area of applications able to fight against bacterial infections. However, almost all antibiotics were baffled by the great ability to adaptation of bacteria (1) and the emergence of new bacterial agents, discovered with up-dated technologies. The living world is perpetually in co-evolution and since more than 3 billion years, bacteria have developed resistance mechanisms to overcome external aggressions. Thus, in the middle of the 80th century, multi-resistant bacteria appeared and disseminated out from hospitals. In this context, researches have been developed in order to find new antimicrobial substances to destroy such new types of bacteria. Thus, several groups have turned their focus on invertebrates, which co evoluad with human and have appeared on the planet since a long time. Evidence of new families of antimicrobial substances isolated from invertebrates different to the classical cationic peptide family i.e. dipeptides and anionic peptides been given. Moreover, these molecules are also present in human and may serve in the innate immune response as an important survival strategy. PMID- 16375701 TI - Syntheses of hydroxylated cyclic beta-amino acid derivatives. AB - This review is intended to give a short summary of the developments in the field of natural and synthetic alicyclic and heterocyclic hydroxylated beta-amino acids and to focus on the main strategies that have been reported for their synthesis. Given the medicinal and biological significance of the hydroxylated beta-amino acids, an increasing volume of research is currently being directed toward regio , stereo- and enantioselective access to this class of compounds. PMID- 16375702 TI - A yeast extract high in bioactive peptides has a blood-pressure lowering effect in hypertensive model. AB - It was reported that angiotensin-converting enzyme (ACE) plays an important role in increasing blood pressure. Recently, it was reported that several food hydrolysates have ACE inhibitory effects in the spontaneous hypertensive rat (SHR) model and mildly hypertensive subjects. Therefore, the anti-hypertensive effects of brewer's yeast BY-G were investigated, which contains many kinds of beneficial nutrients (vitamins, minerals, nucleic acids, glutathione, amino acids, etc.). The aim of this study was to evaluate the anti-hypertensive effects of BY-G and its component peptides obtained by enzymatic treatment. The peptide fraction KRF814 was obtained by the hydrolysate of BY-G with alkaline protease and then treated with Amberlite XAD-2. The KRF814, which has an inhibitory effect on ACE in vitro, was obtained. BY-G and KRF814 were fed to male SHR and showed significantly anti-hypertensive effects. KRF814 contained alanyl-phenylalanine (AF) and glycyl-phenylalanine (GF), which significantly decreased systolic BP in the SHR model. The active ingredients of KRF814, AF, and GF had about 60% of the potency of the positive control, which was captopril. It is considered that intake of BY-G or its component peptides as a functional food stuff might be beneficial for improving BP in people with hypertension. PMID- 16375703 TI - Telomerase inhibition and cancer: might platinum based drugs have a future as anti-telomerase pharmacological approach? AB - Telomerase is a ribonucleoprotein polymerase that maintains the length of telomeric DNA by adding hexameric units (TTAGGG) to the ends of the chromosomes. This mechanism prevents replicative senescence, thus conferring unlimited proliferative potential to cells. Telomerase reactivation has been detected in most human tumour tissue, indicating that the enzyme may be useful as a specific tumour marker. The inhibition of telomerase causes a progressive and critical reduction of telomeres, leading to a potent signal for the blockage of cell proliferation and the induction of apoptosis. Since normal somatic cells lack telomerase activity, the anti-telomerase approach is highly specific for tumour cells and metastases. Prolonged treatment is required before enzyme deactivation causes the telomeres to be shortened enough to induce senescence and apoptosis. Therefore, the drugs employed in anti-telomerase therapy should be of only moderate non-specific cytotoxicity. Certain cis-Pt(II)-complexes have recently been shown to be effective inhibitors of telomerase in both cell-free and in vitro assays, most likely by targeting the nucleobases of the RNA component of the enzyme. PMID- 16375704 TI - Quorum sensing inhibition: targeting chemical communication in gram-negative bacteria. AB - Quorum sensing (QS) systems comprise a new therapeutic target potentially substitutive or complementary to traditional antibiotic treatment of chronic diseases. One route to disrupt the previously established interrelationship between pathogenesis and QS is by blocking the dual functioning signal/receptor transcriptional regulator in some clinically relevant Gram-negative bacteria. The present review contains all reported compound types that are currently known to inhibit the QS transcriptional regulator in Gram-negative bacteria. These compounds are sub-divided into two main groups, one comprising structural analogs of the native signaling molecules and the other compounds lacking structural resemblance. Biological activity is rationalized on the basis of structure activity relationships and structural insight into the target protein. PMID- 16375705 TI - The assessment of platelet activation in antiplatelet drug development. AB - Platelet activation plays an important role in a wide range of pathological conditions. For example, platelet activation has been shown to be involved in the defence against parasitic infection, the pathogenesis of atherosclerotic disease, and various arterial and venous thrombotic diseases. Indeed, there is considerable interest in the manipulation of platelet function for therapeutic gain. It is for these reasons that there is considerable interest in developing assays measuring in vivo platelet activation. Current modalities in the measurement of platelet activation include Enzyme-linked Immunosorbent Assays (ELISA), platelet flow cytometry and electron microscopy. It is proposed that methods in measuring platelet activation can also be classified into 'direct' and 'indirect' modalities, both of which have their distinct advantages and disadvantages. Unfortunately, there is at present no consensus on the ideal method of measuring platelet activation. Thus, studies on platelet activation should ideally include at least one of each of direct and indirect modality of studying platelet activation. This review provides an overview of basic platelet biology and the various methods of measuring platelet activation, with an emphasis on their role in drug development. PMID- 16375706 TI - Phosphate isosteres in medicinal chemistry. AB - The phosphate group is at the heart of an enormous number of biological processes. The simple phosphorylation or dephosphorylation of a protein can have a wide range of consequences, including effects on its biological activity, its interaction with other proteins, and on its subcellular location. Abnormal levels of protein phosphorylation have been linked to a wide range of diseases including cancer and diabetes. Consequently, proteins that recognise the phosphate moiety have become an attractive target for therapeutic development. The most prevalent medicinal chemistry research examines the interactions of phosphorylated tyrosine residues; however, the role of phosphate groups on serine or threonine residues, in nucleotides, DNA and RNA, on sugars, and lipid mediators such as lysophosphatidic acid should not be overlooked. Investigations have focused on the non-catalytic phosphotyrosine-recognising domains such as Src homology 2 (SH2) and phosphotyrosine binding (PTB) domains, as well as catalytic proteins such as protein tyrosine phosphatase 1B (PTP1B). The utilisation of the phosphate moiety as part of an inhibitor is severely limited by the enzymatic lability and poor cellular bioavailability of this highly charged recognition element. The development of phosphate isosteres attempts to address these issues by introducing a non-scissile bond and utilizing groups with less charge that are still able to interact favourably with the target protein in much the same way as the phosphate group does. Many phosphate mimics retain the phosphorus atom such as in the highly successful fluoromethylenephosphonates, whereas others have lost the tetrahedral phosphate geometry and are based on the combination of one or more carboxylate groups that generally reduce the overall charge of the molecule. This review focuses on the recent developments and the use of phosphate isosteres in medicinal chemistry, covering roughly the past four years. PMID- 16375707 TI - RNA interference as a gene-specific approach for molecular medicine. AB - The discovery of RNA interference (RNAi) in eukaryotic cells has been the major recent breakthrough in molecular and cell biology. RNAi machineries exert biological functions in gene regulation, genome defense and chromatin architecture and dynamics. The potential of RNAi to silence any gene of interest in a highly specific and efficient manner via double-stranded RNA (dsRNA) has literally revolutionized modern genetics. RNAi-based functional genomics now permits, for the first time, to evaluate the cellular role of individual gene products on a genome-wide scale in higher organisms like mammals, presenting an alternative to the generation of animal knockouts often doomed to failure because of a lethal phenotype. RNAi has had an enormous impact on the development of novel disease models in animals, and it is likely that small interfering RNAs (siRNAs), which are the trigger molecules for RNA silencing, will become an invaluable tool for the treatment of genetic diseases. First clinical trials, using siRNAs directed against the vascular endothelial growth factor (VEGF) or one of its receptors, have been initiated recently for the treatment of age related macular degeneration. Improving guidelines for the rational design of siRNAs, based on recent progress in understanding the mechanisms underlying RNAi, as well as the introduction of chemical modifications into siRNAs are expected to improve their pharmacokinetic and pharmacodynamic properties for in vivo applications. Finally, successful therapeutic application of RNAi will depend on the development of improved siRNA delivery strategies that combine high specificity and efficiency with a low immunostimulatory and tumorigenic potential. PMID- 16375708 TI - Emerging viruses: advances and challenges. PMID- 16375709 TI - Novel antiviral strategies to combat human Arenavirus infections. AB - Arenaviruses merit significant attention both as tractable model systems to study acute and persistent viral infections, and as clinically important human pathogens. Evidence indicates that LCMV remains present in the USA and Europe and capable of causing significant morbidity in infected individuals, likely being a neglected human pathogen. Moreover, new arenaviruses are being discovered in the Americas on the average of one every three years, with some of them causing severe hemorrhagic fever. In addition, weaponized forms of these viruses pose a real threat as agents of bioterrorism. Therefore, it is important to develop effective vaccines and better antiviral drugs to combat the dual threats of naturally occurring and intentionally introduced Arenavirus infections. The development of arenavirus reverse genetic systems is allowing investigators to conduct a detailed molecular characterization of the viral cis-acting signals and trans-acting factors that control each of the steps of the Arenavirus life cycle, including RNA synthesis, packaging and budding. We will discuss how this new knowledge is facilitating the establishment of novel assays to identify and characterize compounds capable of interfering with specific steps of the virus life cycle. Likewise, the ability to generate predetermined specific mutations within the arenavirus genome, and analyze their phenotypic expression, would significantly contribute to the elucidation of arenavirus-host interactions, including the bases of their ability to persist, as well as to cause severe HF (hemorrhagic fever) disease in humans. These approaches could also lead to the development of novel potent and safe Arenavirus vaccines. PMID- 16375710 TI - Crimean-Congo hemorrhagic fever virus. AB - Crimean-Congo hemorrhagic fever virus (CCHFV) is an important human pathogen, which is the cause of a tick-borne illness occurring in many areas of Africa, Asia, and Europe. CCHF is characterized by a sudden onset of high fever, chills, and severe headache. Other symptoms can include gastrointestinal disorders, such as nausea, vomiting, and diarrhea. In severe cases, hemorrhagic manifestations can occur and often present as large areas of ecchymosis, rather than frank bleeding. Exposure to ticks, particularly those in the genus Hyalomma, or direct contact with virus-infected animals or people are considered the major risk factors. Studies on CCHFV are impeded by the biocontainment needed for their manipulation. However, the increasing worldwide medical awareness, the enormous interest of the media in hemorrhagic fever diseases, and their potential to be used as a bioweapon, have greatly spurred on research on this important virus, as evidenced by many new developments including the development of a reverse genetics system which should greatly enhance future research with this virus. PMID- 16375711 TI - Ebola and Marburg viruses: pathogenesis and development of countermeasures. AB - Ebola and Marburg viruses, family Filoviridae, are among the best known examples of emerging and re-emerging pathogens. Although outbreaks have been sporadic and geographically restricted to areas of Central Africa, the hemorrhagic fevers caused by these viruses are remarkably severe and are associated with high case fatality rates often exceeding 80 percent. In addition to humans, these viruses have decimated populations of wild apes in Central Africa. Currently, there are no vaccines or effective therapies available for human use. Progress in understanding the geneses of the pathophysiological changes that make filoviral infections of humans so destructive has been slow, primarily because these viruses require special containment for safe research. However, an increasing understanding of the molecular mechanisms of filoviral pathogenesis, facilitated by the development of new tools to elucidate critical regulatory elements in the viral life cycle, is providing new targets that can be exploited for therapeutic interventions. In addition, substantial progress has been made in developing recombinant vaccines against these viruses. PMID- 16375712 TI - Hantaviruses: molecular biology, evolution and pathogenesis. AB - Hantaviruses are tri-segmented negative sense single stranded RNA viruses that belong to the family Bunyaviridae. In nature, hantaviruses are exclusively maintained in the populations of their specific rodent hosts. In their natural host species, hantaviruses usually develop a persistent infection with prolonged virus shedding in excreta. Humans become infected by inhaling virus contaminated aerosol. Unlike asymptomatic infection in rodents, hantaviruses cause two acute febrile diseases in humans: hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). The mortality rate varies from 0.1% to 40% depending on the virus involved. Hantaviruses are distributed world wide, with over 150,000 HFRS and HPS cases being registered annually. In this review we summarize current knowledge on hantavirus molecular biology, epidemiology, genetic diversity and co-evolution with rodent hosts. In addition, special attention was given in this review to describing clinical manifestation of HFRS and HPS, and advances in our current understanding of the host immune response, treatment, and prevention. PMID- 16375713 TI - Emerging influenza viruses: past and present. AB - Influenza is an example of a disease for which the viral pathogen has emerged into the human population many times over past centuries, sometimes with devastating consequences [1]. Historical records provide vivid descriptions of past influenza outbreaks, and the viruses that caused the pandemics of the last century remain subjects of great interest. It is almost certain that a new pandemic, caused by the zoonotic transmission of a new influenza virus into humans, will occur. The recent outbreaks of the highly pathogenic H5 and H7 subtype viruses in poultry and their limited transmission into humans, as well as transmission of H9 subtype viruses, have raised concerns that conditions are developing for the generation of a new pandemic virus. In this paper, we review past pandemics, viral determinants of cross-species transmission of viruses, molecular factors that contribute to disease, and preventative measures to reduce the impact of a future pandemic. PMID- 16375714 TI - Hendra and Nipah viruses: pathogenesis and therapeutics. AB - Within the past decade a number of new zoonotic paramyxoviruses emerged from flying foxes to cause serious disease outbreaks in man and livestock. Hendra virus was the cause of fatal infections of horses and man in Australia in 1994, 1999 and 2004. Nipah virus caused encephalitis in humans both in Malaysia in 1998/99, following silent spread of the virus in the pig population, and in Bangladesh from 2001 to 2004 probably as a result of direct bat to human transmission and spread within the human population. Hendra and Nipah viruses are highly pathogenic in humans with case fatality rates of 40% to 70%. Their genetic constitution, virulence and wide host range make them unique paramyxoviruses and they have been given Biosecurity Level 4 status in a new genus Henipavirus within the family Paramyxoviridae. Recent studies on the virulence, host range and cell tropisms of henipaviruses provide insights into the unique biological properties of these emerging human pathogens and suggest approaches for vaccine development and therapeutic countermeasures. PMID- 16375715 TI - Countermeasures to the bioterrorist threat of smallpox. AB - Variola, the agent of smallpox, is a bioterrorist threat, as is monkeypox virus, which also occurs naturally in Africa. Development of countermeasures, in the form of improved vaccines, antiviral drugs, and other therapeutic strategies are a high priority. Recent advances in molecular biology and in animal model development have provided fresh insight into the virulence determinants for smallpox and the pathophysiology of disease. The complex replication cycle for orthopoxviruses, and the pivotal role for viral-specific immunomodulatory proteins which contribute to escape from immunologic surveillance, provide many unique targets for therapeutic intervention. The "toxemia" of smallpox has been elucidated in part by variola-infected primate studies which revealed the central role of apoptosis and the evolution of a cytokine storm leading to hemorrhagic diathesis, resembling fulminent "black" smallpox. This suggests a potential role for therapeutic strategies developed for septic shock, in treatment of smallpox. Drugs licensed for other viruses which share molecular targets with orthopoxviruses (e.g. Cidofovir) or cancer drugs (e.g. Gleevec and other tyrosine kinase inhibitors) have immediate application for treatment of smallpox and monkeypox and provide leads for second generation drugs with higher therapeutic indices. Recent advances in identification of virulence determinants and immune evasion genes facilitate the design of alternative vaccines to replace live vaccinia strains that are unsuitable for a large proportion of individuals in a mass immunization campaign. PMID- 16375716 TI - Rift Valley fever virus. AB - Rift Valley fever is considered to be one of the most important viral zoonoses in Africa. In 2000, the Rift valley fever virus spread to the Arabian Peninsula and caused two simultaneous outbreaks in Yemen and Saudi Arabia. It is transmitted to ruminants and to humans by mosquitoes. The viral agent is an arbovirus, which belongs to the Phlebovirus genus in the Bunyaviridae family. This family of viruses comprises more than 300 members grouped into five genera: Orthobunyavirus, Phlebovirus, Hantavirus, Nairovirus, and Tospovirus. Several members of the Bunyaviridae family are responsible for fatal hemorrhagic fevers: Rift Valley fever virus (Phlebovirus), Crimean-Congo hemorrhagic fever virus (Nairovirus), Hantaan, Sin Nombre and related viruses (Hantavirus), and recently Garissa, now identified as Ngari virus (Orthobunyavirus). Here are reviewed recent advances in Rift Valley fever virus, its epidemiology, molecular biology and focus on recent data on the interactions between viral and cellular proteins, which help to understand the molecular mechanisms utilized by the virus to circumvent the host cellular response. PMID- 16375717 TI - Recent advances in the molecular biology of west nile virus. AB - Since the mid-1990s, West Nile virus (WNV) has emerged as a significant agent of arboviral encephalitis in several regions of the world. In 1999, WNV was introduced into the northeastern United States and was associated with an outbreak of encephalitis affecting humans, birds and horses. Subsequently, the virus has spread across the country, and across southern Canada, and in 2002 and 2003 was associated with the largest outbreaks of arboviral encephalitis recorded in the Western hemisphere. Interestingly, the more recent spread of WNV into Mexico, Central America and the Caribbean has not been associated with the high levels of clinical disease observed in North America. This review addresses the most recent results from studies investigating the molecular biology and evolution of WNV, as well as progress in the development of diagnostic and therapeutic reagents. PMID- 16375718 TI - Aging blunts ischemic-preconditioning-induced neuroprotection following transient global ischemia in rats. AB - The present study examines the hypothesis that aging defined by the 50% survival age compromises neuroprotection afforded by ischemic preconditioning (IPC). Sixty four male F344 rats aged 4- and 24-months, respectively, were subjected to IPC, (3-min ischemia) or sham-surgery followed by 10-min (full) ischemia or sham surgery 2 days later. There were 4 groups at each age: sham-surgery-sham-surgery (SS), preconditioning-sham-surgery (PS), preconditioning-ischemia (PI) and sham surgery-ischemia (SI) groups. Assessments of histology and immunoreactivities of N-methyl-D-aspartic acid receptor 1 (NMDAr1) and caspase-3 active peptide (C3AP) in the hippocampal CA1 region were performed 8 days after full ischemia. The CA1 "living cell ratio" was greater in the aged SI group than in the young SI group (32+/-6% vs. 17+/-5%, p<0.05), whereas the degree of protection against full ischemia afforded by IPC was reduced in the aged compared with the young (53+/ 17% vs. 241+/-25%, P<0.0001). The basal level of NMDAr1 immunofluorescence was significantly higher in young animals, while the numbers of C3AP-positive cells were greater in all three aged ischemic groups as compared to respective young groups (p<0.01, p=0.055 and p<0.05). A fourth method of assessing cell damage using Fluoro Jade C labeled degenerating neurons that were also intensively eosinophilic. Counts of Fluoro Jade C-positive cells were higher in the young SI group than in the aged SI group (P<0.05), suggesting that mechanisms of ischemic cell death may change with aging. In conclusion, aging alters mechanisms of ischemic cell death in CA1 neurons and ischemic tolerance mechanisms are blunted by aging. PMID- 16375719 TI - Hyperglycemia potentiates carbonyl stress-induced apoptosis in naive PC-12 cells: relationship to cellular redox and activator protease factor-1 expression. AB - The mechanism(s) of central nervous system complication associated with neurodegenerative disorders such as diabetes is unknown. Previous studies demonstrated that carbonyl stress induced by methylglyoxal (MG) mediates differential apoptosis of rat pheochromocytoma (PC12) cells in the naive or differentiated transition states. Since chronic hyperglycemia is central to diabetic complications, and poorly differentiated cells are oxidatively more vulnerable, we currently investigated the effect of glycemic status on MG-induced apoptosis in naive (nPC12) cells focusing on glutathione-to-glutathione disulfide (GSH/GSSG) redox signaling. nPC12 cells were exposed to 25 mM glucose acutely for 24h or chronically for 1 week. A role for glycemic fluctuation was tested in chronic high glucose-adapted cells subjected to acute reduction in glucose availability. Acute hyperglycemia potentiated MG-induced nPC12 apoptosis in accordance with cellular redox (GSH-to-Disulfide (GSSG plus protein-bound SSG)) imbalance. Chronic hyperglycemia exacerbated baseline and MG-induced apoptosis that corresponded to exaggerated loss of cytosolic and mitochondrial redox balance, impaired glucose 6-phosphate dehydrogenase (G6PD) activity, and enhanced basal expression of apoptosis protease activator factor-1 (Apaf-1). Reduced glucose availability in hyperglycemia-adapted nPC12 cells induced by acute lowering of glucose or by dehydroepiandrosterone (DHEA, G6PD inhibitor) further enhanced MG-induced apoptosis in association with greater cytosolic and mitochondrial redox and G6PD impairment and elevated basal Apaf-1 expression. These findings demonstrate that chronic hyperglycemia or acute glucose reduction from the chronic hyperglycemic state potentiates carbonyl stress, which collectively contribute to oxidative susceptibility of poorly differentiated cells such as that which occurs in brain neurons of neurodegenerative disorders like diabetes and Alzheimer's disease. PMID- 16375721 TI - Increased behavioral and histological variability arising from changes in cerebrovascular anatomy of the Mongolian gerbil. AB - The Mongolian gerbil (Meriones unguiculatus) has been used extensively as a model of forebrain ischemia. Its unique susceptibility to ischemia was suggested to be due to an incomplete circle of Willis. The relative ease to which ischemia can be induced combined with highly reproducible delayed CA1 cell death following a 5 min occlusion made the model popular in neuroprotection studies. Presently, this assumption was tested that complete forebrain ischemia occurs in all gerbils because increased variability was noticed in neuronal injury and behavioral outcome using this model in the last several years. Here it is reported that gerbils obtained from Charles River, the largest supplier in North America, show a high incidence (22.7% with bilateral and 38.6% with unilateral anastomoses) of posterior communicating arteries compared to another supplier of gerbils (High Oak Farms, 2.6% with bilateral and 13.2% with unilateral anastomoses, P<0.0001). This increased incidence of complete or partial circle of Willis led to less severe CA1 cell loss in Charles River gerbils (P<0.0001) compared to High Oak gerbils, with an unacceptably high level of inter-animal variability. Similarly, behavioral indices of CA1 ischemic injury (increased locomotion, habituation deficits) were also significantly attenuated in the Charles River animals. High Oak gerbils also displayed increased histological and behavioral variability relative to the pattern obtained several years ago. Thus, the gerbil model of forebrain ischemia, at least using Charles River animals, no longer produces consistent injury and behavioral alterations. Investigators are urged to consider adopting other models in future neuroprotection studies or ensure that their gerbil population lacks communicating arteries. PMID- 16375720 TI - Erythropoietin requires NF-kappaB and its nuclear translocation to prevent early and late apoptotic neuronal injury during beta-amyloid toxicity. AB - No longer considered exclusive for the function of the hematopoietic system, erythropoietin (EPO) is now considered as a viable agent to address central nervous system injury in a variety of cellular systems that involve neuronal, vascular, and inflammatory cells. Yet, it remains unclear whether the protective capacity of EPO may be effective for chronic neurodegenerative disorders such as Alzheimer's disease (AD) that involve beta-amyloid (Abeta) apoptotic injury to hippocampal neurons. We therefore investigated whether EPO could prevent both early and late apoptotic injury during Abeta exposure in primary hippocampal neurons and assessed potential cellular pathways responsible for this protection. Primary hippocampal neuronal injury was evaluated by trypan blue dye exclusion, DNA fragmentation, membrane phosphatidylserine (PS) exposure, and nuclear factor kappaB (NF-kappaB) expression with subcellular translocation. We show that EPO, in a concentration specific manner, is able to prevent the loss of both apoptotic genomic DNA integrity and cellular membrane asymmetry during Abeta exposure. This blockade of Abeta generated neuronal apoptosis by EPO is both necessary and sufficient, since protection by EPO is completely abolished by co-treatment with an anti-EPO neutralizing antibody. Furthermore, neuroprotection by EPO is closely linked to the expression of NF-kappaB p65 by preventing the degradation of this protein by Abeta and fostering the subcellular translocation of NF-kappaB p65 from the cytoplasm to the nucleus to allow the initiation of an anti-apoptotic program. In addition, EPO intimately relies upon NF-kappaB p65 to promote neuronal survival, since gene silencing of NF-kappaB p65 by RNA interference removes the protective capacity of EPO during Abeta exposure. Our work illustrates that EPO is an effective entity at the neuronal cellular level against Abeta toxicity and requires the close modulation of the NF-kappaB p65 pathway, suggesting that either EPO or NF-kappaB may be used as future potential therapeutic strategies for the management of chronic neurodegenerative disorders, such as AD. PMID- 16375722 TI - The function and integrity of the neurovascular unit rests upon the integration of the vascular and inflammatory cell systems. AB - The neurovascular unit is composed of a microvascular endothelium, neuron, and glial cell elements that are in physical proximity to the endothelium. The vascular system provides oxygen, glucose, and hormones for brain cells and guides the cells to appropriately respond to the local environment. Conversely, the brain cells, especially glial cells, can regulate the function of blood vessels in response to local requirements. The disruption of the neurovascular coordination was observed in a variety of inflammation-related diseases in brain, such as infectious diseases, stroke, vascular dementia, and multiple sclerosis. Inflammatory responses resulting from infections or injury of the brain activate the endothelium and glial cells to various degrees depending on the type, titer, or strength and duration of exposure to the agents or insults. The activation of endothelial and microglial cells may be modulated by the action of cytokines or other substances secreted from these cells. In an effort to understand the pathogenesis and find rational treatments against inflammatory disorders in brain, studies have been separately carried out using either endothelial cells or microglia. Increasing evidence, however, indicates that a crosstalk between these two cell types is important for the brain inflammation. Here, we review recent advances that provide insights into the coordinated interaction between the vascular and microglial systems, including the role of the specialized endothelium in regulating the immune response that occurs within CNS, the influence of microglial cells on the properties of endothelial cells, and the effects of endothelium on the state of microglial activation. PMID- 16375724 TI - The emerging role of coenzyme Q-10 in aging, neurodegeneration, cardiovascular disease, cancer and diabetes mellitus. AB - Coenzyme Q (ubiquinone, 2-methyl-5,6-dimethoxy-1,4-benzoquinone), soluble natural fat quinine, is crucial to optimal biological function. The coenzyme Q molecule has amphipathic (biphasic) properties due to the hydrophilic benzoquinone ring and the lipophilic poly isoprenoid side-chain. The nomenclature of coenzyme Q-n is based on the amount of isoprenoid units attached to 6-position on the benzoquinone ring. It was demonstrated that coenzyme Q, in addition to its role in electron transport and proton transfer in mitochondrial and bacterial respiration, acts in its reduced form (ubiquinol) as an antioxidant. Coenzyme Q 10 functions as a lipid antioxidant regulating membrane fluidity, recycling radical forms of vitamin C and E, and protecting membrane phospholipids against peroxidation. The antioxidant property, high degree of hydrophobicity and universal occurrence in biological system, suggest an important role for ubiquinone and ubiquinol in cellular defense against oxidative damage. Coenzyme Q 10 is a ubiquitous and endogenous lipid-soluble antioxidant found in all organisms. Neurodegenerative disorders, cancer, cardiovascular diseases and diabetes mellitus and especially aging and Alzheimer's disease exhibit altered levels of ubiquinone or ubiquinol, indicating their likely crucial role in the pathogenesis and cellular mechanisms of these ailments. This review is geared to discuss the biological effect of coenzyme Q with an emphasis on its impact in initiation, progression, treatment and prevention of neurodegenerative, cardiovascular and carcinogenic diseases. PMID- 16375726 TI - Diversity in the activity of individual enzymes. AB - Although the structure of an enzyme is often depicted as static, it is dynamic. Hence, a population of chemically identical enzymes has not one, but a distribution of structures at any moment in time. Does this have an effect on the activity of the enzyme? This article reviews experiments designed to test the hypothesis that this distribution of structures results in a distribution of enzyme activities. The experiments reviewed here use different enzymes, falvin adenine dinucleotide, beta-galactosidase, alkaline phosphatase, exonuclease I, lactate dehydrogenase I, alpha-chymotrypsin, the 20S proteasome, and horseradish peroxidase. All experiments come to the same conclusion, when measured individually, apparently identical enzymes show a distribution in rates of activity. PMID- 16375727 TI - Colloidal behavior of proteins: effects of the second virial coefficient on solubility, crystallization and aggregation of proteins in aqueous solution. AB - There has been an increasing awareness that proteins, like other biopolymers, are large enough to exhibit colloidal behavior in aqueous solution. Net attractive or repulsive forces have been found to govern important physical properties, such as solubility and aggregation. The extent of intermolecular interactions, usually expressed in terms of the osmotic second virial coefficient, B, is most often measured using static light scattering. More recently, self-interaction chromatography (SIC) has emerged as a method for rapid determination of B in actual formulations, as it uses much less protein and has higher throughput. This review will summarize the relationship of B to crystallization, solubility, and aggregation of proteins in aqueous solution. Moreover, the capability of SIC to obtain B values in a rapid and reproducible fashion will be described in detail. Finally, the use of miniaturized devices to measure B is presented. PMID- 16375728 TI - How the molecule number is correctly quantified in two-color fluorescence cross correlation spectroscopy: corrections for cross-talk and quenching in experiments. AB - Fluorescence correlation spectroscopy (FCS) and two-color fluorescence cross correlation spectroscopy (FCCS) are among the cutting-edge technologies for measuring molecule numbers at the single-molecule level in liquid phases. Yet, even after single molecule technologies caught up with theory, the techniques remained tools only for specialists able to navigate the formulas that give meaning to their observations. This original article aims at the derivations of relevant and useful quantification of molecule numbers for researchers with more diverse backgrounds who have begun probing questions previously unanswerable, except on the level of the molecule. The quantitation depends on the exact conditions of measurement. To some extent these are arbitrary, so that standard procedures are necessary in for valid comparisons of measurements among different data sets. To agree on and specify such procedures is one of the further aims here. No matter what fluorophores, which have, of course, to meet photophysical and photochemical requirements for FCS/FCCS, and optical setups/devices are used, the primary measurement signal arises from fluctuations of the mean molecule number in a confocal femtoliter or smaller probe region. Since FCS/FCCS relies on fluorescence emission measurements of rare events, one is looking for small signals on essentially zero background. Optical separation by FCCS setups is usually defined in terms of cross-talk and cross-excitation/cross-emission, respectively, which can be calculated and minimized by the experimenter from readily measurable quantities of the absorption/emission scenario for single labels and multiple labels n and m bound to or incorporated into the two-color molecules. Furthermore, this article derives relevant formulas for the quantification of molecule numbers under different experimental conditions with substantial quenching of the two-color molecules such as single labels and multiple labels n and m bound to or incorporated into the two-color molecules, high-density labeling of two-color molecules with multiple n green labels and one red label. Here, we summarize and extend the formulas to make them more generally applicable. PMID- 16375729 TI - Monitoring nucleic acids using molecular beacons. AB - The ability to observe the dynamic of RNA in single living cells offers many exciting opportunities in biology and medicine. In the last few years, molecular beacons (MBs) have shown great potential in monitoring RNA synthesis, transportation, and localization with good sensitivity and selectivity. A hairpin structure probe, MB is a dual-labeled single stranded oligonucleotide that only fluoresces in the presence of target sequences. In this paper, the basic principle and design of MB will be described. The application of MB for RNA imaging in living cells will be reviewed. The limitations of MB for in vivo application will be identified. In the last section of the article, the efforts on designing better MBs for highly sensitive and selective RNA imaging will be discussed. PMID- 16375723 TI - Driving cellular plasticity and survival through the signal transduction pathways of metabotropic glutamate receptors. AB - Metabotropic glutamate receptors (mGluRs) share a common molecular morphology with other G protein-linked receptors, but there expression throughout the mammalian nervous system places these receptors as essential mediators not only for the initial development of an organism, but also for the vital determination of a cell's fate during many disorders in the nervous system that include amyotrophic lateral sclerosis, Parkinson's disease, Alzheimer's disease, Huntington's disease, Multiple Sclerosis, epilepsy, trauma, and stroke. Given the ubiquitous distribution of these receptors, the mGluR system impacts upon neuronal, vascular, and glial cell function and is activated by a wide variety of stimuli that includes neurotransmitters, peptides, hormones, growth factors, ions, lipids, and light. Employing signal transduction pathways that can modulate both excitatory and inhibitory responses, the mGluR system drives a spectrum of cellular pathways that involve protein kinases, endonucleases, cellular acidity, energy metabolism, mitochondrial membrane potential, caspases, and specific mitogen-activated protein kinases. Ultimately these pathways can converge to regulate genomic DNA degradation, membrane phosphatidylserine (PS) residue exposure, and inflammatory microglial activation. As we continue to push the envelope for our understanding of this complex and critical family of metabotropic receptors, we should be able to reap enormous benefits for both clinical disease as well as our understanding of basic biology in the nervous system. PMID- 16375730 TI - Single-molecule detection and probe strategies for rapid and ultrasensitive genomic detection. AB - This paper reviews the current state-of-the-art development of single-molecule detection (SMD)-based methods for ultrasensitive and specific analysis of genomic sequences. We first discuss several newly devised single fluorescent probe strategies that allow separation-free detection of low-abundance DNA sequences, such as quantum dot (QD)-mediated fluorescence resonance energy transfer (FRET) technology and dual-color fluorescence coincidence and colocalization analysis. Various schemes toward single DNA sizing and sequencing in solutions or on surfaces are also reviewed. In the end, we summarize the different microfluidic approaches developed for use with SMD to facilitate rapid, low-volume and quantitative analysis, such as electrokinetic and hydrodynamic single-molecule manipulation techniques. PMID- 16375732 TI - Overview of hyperuricaemia and gout. AB - In most mammals purine degradation ultimately leads to the formation of allantoin. Humans lack the enzyme uricase, which catalyzes the conversion of uric acid to allantoin. The resulting higher level of uric acid has been hypothesized to play a role as an antioxidant. Hyperuricaemia is usually an asymptomatic condition which is hypothesized to play a role in cardiovascular disease and hypertension. Some hyperuricaemic individuals develop gout, an inflammatory arthritis caused by the deposition of monosodium urate crystals in joints. Over time, acute intermittent gouty arthritis can develop into a chronic condition with deposits of monosodium urate (MSU) crystals in joints and as tophi. The mechanisms by which MSU crystals lead to an acute inflammatory arthritis are under investigation and current knowledge is reviewed here. Treatment of gout includes management of acute flares with anti-inflammatory medications such as non-steroidal anti-inflammatory drugs or corticosteroids and long term management with urate-lowering therapy when indicated. Future directions in the treatment of gout, in part guided by a better understanding of pathophysiology, are discussed. PMID- 16375733 TI - Serum uric acid and risk of coronary heart disease. AB - Many large epidemiological studies confirmed a positive association between raised serum uric acid (SUA) levels and risk of coronary heart disease (CHD) or cardiovascular disease (CVD) in the general population, among hypertensive patients and those with established CHD, stroke, diabetes and heart failure. There is much controversy concerning the role of SUA as an independent risk factor for CHD as SUA is related to many of the established risk factors for cardiovascular disease including hypertension, dyslipidaemia, obesity and pre existing disease. The epidemiological evidence suggests that SUA is an independent predictor of CVD in subjects with hypertension and established vascular disease but not in healthy subjects. This evidence suggests that the influence of SUA on CHD is explained by the secondary association of SUA with other established etiological risk factors (hypertension, dyslipidaemia, hyperinsulinaemia, obesity and pre-existing disease). There is no evidence so far to indicate that lowering SUA levels with drug treatment has a beneficial effect on CVD outcome. In summary, there is little support for an independent causal role for SUA in the development of CHD. However, SUA may provide useful prognostic information in subjects with hypertension and vascular disease. PMID- 16375734 TI - Dietary factors and hyperuricaemia. AB - The connection of gout and hyperuricaemia with gluttony, overindulgence in food and alcohol and obesity dates from ancient times. Studies from different parts of the world suggest that the incidence and severity of hyperuricaemia and gout may be increasing. Uric acid (urate) is the end product of purine degradation. Although most uric acid is derived from the metabolism of endogenous purine, eating foods rich in purines contributes to the total pool of uric acid. Sustained hyperuricaemia is a risk factor for acute gouty arthritis, chronic tophaceous gout, renal stones and possibly cardiovascular events and mortality. Before starting lifelong urate-lowering drug therapy, it is important to identify and treat underlying disorders that may be contributing to hyperuricaemia. It is relevant to recognize the strong association of the insulin resistance syndrome (IRS) (abdominal obesity, dyslipidaemia, hypertension, raised serum insulin levels and glucose intolerance) with hyperuricaemia. Consumption of meat, seafood and alcoholic beverages in moderation and attention to food portion size is important. Moderation in the consumption of not only beer but also other forms of alcohol is essential. In the obese, controlled weight management has the potential to lower serum urate in a quantitatively similar way to relatively unpalatable "low purine" diets. Non-fat milk and low-fat yogurt have a variety of health benefits and dairy products may have clinically meaningful antihyperuricaemic effects. In addition, fruits, such as cherries and high intakes of vegetable protein diet may reduce serum urate levels. PMID- 16375735 TI - Uric acid and hypertension. AB - Increased levels of uric acid are associated with cardiovascular disease and the metabolic syndrome. They may predict clinical outcomes and also the onset of hypertension, though it is less clear that hyperuricaemia can be regarded as an independent risk factor given its clustering with other well-recognised factors. Uric acid may increase as a result of pathophysiological processes such as impaired renal sodium handling but may also contribute to renal and vascular damage, particularly endothelial dysfunction. It is notable that the synthesis of uric acid may be associated with the generation of reactive oxygen species if the enzyme xanthine oxidorectase is converted to the oxidase, as may occur in ischaemia. It has been suggested that uric acid may play a role in the pathogenesis of early-onset hypertension but evidence for this is limited. There is also very limited data to suggest that in some circumstances lowering uric acid can lower blood pressure. In the metabolic syndrome, the presence of elevated uric acid concentrations is closely associated with raised triglyceride levels, for reasons that have not been clearly defined. It remains to be seen whether uric acid could or should be considered a specific therapeutic target in cardiovascular disease and especially in hypertension and if so what should be the optimal pharmacological approach to lowering serum urate levels. PMID- 16375736 TI - Uric acid and oxidative stress. AB - Uric acid is the final product of purine metabolism in humans. The final two reactions of its production catalyzing the conversion of hypoxanthine to xanthine and the latter to uric acid are catalysed by the enzyme xanthine oxidoreductase, which may attain two inter-convertible forms, namely xanthine dehydrogenase or xanthine oxidase. The latter uses molecular oxygen as electron acceptor and generates superoxide anion and other reactive oxygen products. The role of uric acid in conditions associated with oxidative stress is not entirely clear. Evidence mainly based on epidemiological studies suggests that increased serum levels of uric acid are a risk factor for cardiovascular disease where oxidative stress plays an important pathophysiological role. Also, allopurinol, a xanthine oxidoreductase inhibitor that lowers serum levels of uric acid exerts protective effects in situations associated with oxidative stress (e.g. ischaemia reperfusion injury, cardiovascular disease). However, there is increasing experimental and clinical evidence showing that uric acid has an important role in vivo as an antioxidant. This review presents the current evidence regarding the antioxidant role of uric acid and suggests that it has an important role as an oxidative stress marker and a potential therapeutic role as an antioxidant. Further well designed clinical studies are needed to clarify the potential use of uric acid (or uric acid precursors) in diseases associated with oxidative stress. PMID- 16375737 TI - Uric acid and the kidney: urate transport, stone disease and progressive renal failure. AB - In this brief review and update, we try to cover recent developments in our understanding of uric acid transport by the kidney, the contribution of uric acid to renal stone disease, its potential role in progressive renal failure and, most recently, the novel and as yet unexplained link between the urinary glycoprotein Tamm-Horsfall protein (uromodulin) and hyperuricaemia and two inherited forms of renal disease with chronic renal failure. PMID- 16375738 TI - Effect on serum uric acid levels of drugs prescribed for indications other than treating hyperuricaemia. AB - Beyond allopurinol and the well-established uricosuric drugs, several other agents can decrease serum uric acid (SUA) levels, such as losartan, fenofibrate and some non-steroidal anti-inflammatory drugs (NSAIDs). Some of these drugs increase renal urate excretion. Hyperuricaemia and gout are common problems (at least 1% of Western men are affected by gout). Raised SUA levels increase the incidence of acute gout and renal calculi. Hyperuricaemia may also predict an increased risk of vascular events. Therefore, lowering SUA levels is of clinical relevance. In this review we consider the effect on SUA levels of drugs that are prescribed for indications other than treating hyperuricaemia. These drugs may obviate the need for specific treatment (e.g. allopurinol) aimed at lowering SUA levels. Furthermore, because hyperuricaemic patients may already be on several drugs (e.g. due to associated dyslipidaemia, hypertension and/or arthritis) compliance may be improved by avoiding additional medication. The potential for adverse effects associated with polypharmacy would also be decreased. PMID- 16375739 TI - Rasburicase: a new approach for preventing and/or treating tumor lysis syndrome. AB - Tumor lysis syndrome (TLS) is an oncologic emergency requiring prompt attention to the management of potentially life-threatening metabolic derangements. Hyperuricaemia is one of the prominent features of TLS which, if not adequately prevented or treated, may lead to renal failure, requiring dialysis. Conventional management of hyperuricaemia involved the use of aggressive hydration, urinary alkalinization and allopurinol. Despite these measures, as many as 14.1% of high risk patients may still develop renal failure. With the advent of newer agents such as rasburicase, the paradigm of TLS management has shifted towards risk stratification and the use of rasburicase in conjunction with hydration in patients at high risk for TLS. The advantage of rasburicase over allopurinol is its rapid onset of action, lack of need for urine alkalinization, which may worsen hyperphosphataemia and a satisfactory safety profile. Overall, rasburicase offers a safe and more effective alternative to allopurinol in patients at highest risk for TLS. Some of the unanswered questions requiring further investigation with regard to rasburicase use include the optimal number of doses needed, optimal dose based on uric acid levels and tumor burden, dosing in obese patients and maximum dose. PMID- 16375740 TI - Raloxifene and cardiovascular health: its relationship to lipid and glucose metabolism, hemostatic and inflammation factors and cardiovascular function in postmenopausal women. AB - CHD is one of the most common serious chronic conditions in postmenopausal women and leads to extremely high risk for recurrent myocardial infarction and death. On the basis of the currently available randomized clinical-trial results the role of conventional HRT for treatment and prevention of CHD is rapidly evolving from presumed benefit to proven harm, at least in some categories of women yet to define. For this reason there has been a particular interest in potential clinical uses of selective estrogen receptor modulators (SERMs). SERMs are a class of compounds that can act as estrogen receptor (ER) agonists in some domains (bone and lipids) and acting as ER antagonists in others (breast and uterus). Raloxifene hydrochloride is an antiestrogen that is currently approved only to treat osteoporosis in postmenopausal women. Because of its effects on lipids and other biomarkers of cardiovascular risk, there is great interest in determining whether it may benefit the cardiovascular system. The great majority of data on cardiovascular effects of raloxifene concern effects on lipids and markers of thrombosis and inflammation. The purpose of this review is to summarize the best available evidence concerning raloxifene and cardiovascular disease focusing some areas known to be important risk factors for cardiovascular diseases: lipids and lipoproteins, glucose metabolism, hemostatic factors, markers of inflammation and cardiovascular function. PMID- 16375741 TI - Recent advances in the discovery of tissue factor/factor VIIa inhibitors and dual inhibitors of factor VIIa/factor Xa. AB - The search for an ideal anticoagulant has spanned decades and has taken several approaches to the identification of novel target molecules for preventing and treating thrombosis. In the group of anticoagulants acting through direct inhibition of coagulation factors, most research has focused on thrombin and factor Xa inhibitors. Attention has been drawn most recently to factor VIIa as a promising anticoagulation target, because of its role in complex with tissue factor, in initiating the coagulation cascade following blood vessel damage. Several reports suggest that inhibitors of the tissue factor/factor VIIa complex prevent thrombosis with a lower bleeding risk than other types of inhibitors. Accordingly, there is increasing interest in the generation of potent and selective small-molecule factor VIIa inhibitors that can be safely administered once or twice daily in an oral formulation with no need for routine coagulation monitoring. The emphasis of this review will be placed on recent advances in the development of the small-molecule inhibitors of factor VIIa complexed with tissue factor. The role of factor VIIa and tissue factor as initiators of the coagulation cascade following blood vessel damage is described, along with the structure of the active site of factor VIIa. PMID- 16375743 TI - Editorial. PMID- 16375742 TI - Lifelong endocrine fluctuations and related cognitive disorders. AB - The aim of this review is to examine the relationship between endocrine fluctuation and cognitive functioning. A plethora of in vitro and in vivo studies has demonstrated the neuroprotective role of estrogens and their impact on the neurotransmitter systems implicated in cognition. Recent hormonal replacement therapy (HRT) trials in non-demented post-menopausal women suggest a temporary positive effect (notably on verbal memory), and four recent meta-analyses converge to suggest a possible protective effect in relation to Alzheimer's disease (reducing risk by 29 to 44%). However, data from the only large randomised controlled trial published to date, the Women's Health Initiative Memory Study, did not confirm these observations and have even suggested an increase in dementia risk for women using HRT compared to controls. Several methodological differences between observation studies and controlled trials with regard to patient group, type, timing and duration of HRT, cognitive measures and analyses, are discussed to explain these discrepancies. The association between hormonal serum level and cognitive functioning remains controversial suggesting high inter-individual vulnerability in risk. Moreover, research on the impact of endocrine functioning on cognition during the female reproductive cycle suggests life-long fluctuations in vulnerability. Etiological models taking into account the interaction of clinical, reproductive, and menstrual events throughout life may provide a more valid approach in understanding the effects of steroids on the brain and in determining sub-groups at heightened risk. Cognitive disorders in the elderly are more likely be related to cumulated lifelong exposure to steroids, rather than to a specific exposure to a given steroid. Multifactorial models based on an exhaustive view of all hormonal events throughout reproductive life together with other risk factors (notably genetic risk factors related to estrogen receptor polymorphisms) should be explored to clarify the role of hormonal risk factors, or protective factors for cognitive dysfunction and dementia. PMID- 16375744 TI - Discovery of next generation inhibitors of HIV protease. AB - Due to factors such as resistance and long-term side effects as well as dosing regimen-related adherence issues, HIV therapy is a constantly moving target. HIV 1 protease inhibitors had an immediate and dramatic impact on the outcome of HIV/AIDS when launched in late 1995, and the search for new and improved next generation molecules has been under way in many laboratories. At GlaxoSmithKline (GSK) and Vertex Pharmaceuticals, this effort focused on two key issues, patient compliance and viral resistance. Using a water-solubilizing prodrug approach, the pill-burden in delivering our protease inhibitor, amprenavir, was dramatically decreased. By eliminating the large amounts of excipients necessary for the original soft-gel formulation, fosamprenavir (Lexiva/Telzir) delivers the clinically efficacious dose of amprenavir with two compact tablets per dose, compared to eight gel capsules. Our efforts to overcome viral resistance to 1(st) generation protease inhibitors by further elaborating the SAR of the amprenavir and related scaffolds, led to successive and dramatic improvements in wild-type antiviral potencies, and ultimately to the discovery of "ultra-potent" molecules with very favorable overall resistance profiles. The selection of GW640385 (brecanvir--USAN approved only) as a clinical candidate and its progression into current phase 2 dose ranging studies represents the culmination of our effort toward next generation protease inhibitors. PMID- 16375745 TI - Recent developments of structure based beta-secretase inhibitors for Alzheimer's disease. AB - The amyloid-beta (Abeta) peptide is the principal components of the senile plaques found in the brains of patients with Alzheimer's disease (AD). The poorly soluble 40-42 amino acid peptide, formed from the cleavage of the Abeta precursor protein (APP) by two proteases, is believed to play a central role in the pathogenesis of AD. Beta-secretase (memapsin 2, BACE1), a membrane-anchored aspartic protease, is responsible for the initial step of APP cleavage leading to the generation of Abeta. Identification and structural determination of beta secretase have established it to be a primary drug target for AD therapy and stimulated active studies on the inhibitors of this protease. Here we review more recent developments in the design and testing of structure-based beta-secretase inhibitors. PMID- 16375746 TI - Inhibitors of dipeptidyl peptidase IV--recent advances and structural views. AB - Prevalence of type 2 diabetes has increased dramatically in the last decades. Current medicines are not yet capable to efficiently prevent or reverse progression of the disease and its associated comorbidities. As a consequence, there is a great need for novel antidiabetic drugs. Treatments of type 2 diabetes that are based on enhanced and sustained action of insulinotropic incretin hormones such as GLP-1 have received much attention in the past years. Treatment strategies include administration of: 1) GLP-1 analogues that are resistant to degradation by the serine protease DPP-IV, and 2) small molecule DPP-IV inhibitors that are able to provide sustained action of endogenous GLP-1, again by preventing its degradation. This review summarizes recent research results for the second approach. It briefly touches upon the advantages that treatment of type 2 diabetes with DPP-IV inhibitors may offer over current medications. In the main section, several important structural classes of DPP-IV inhibitors are described and compared based on literature data. Specific attention is given to the analysis of several X-ray structures of enzyme-inhibitor co-crystals. Finally, as clinical data are steadily emerging for some of the most advanced development candidates, the last section of this review is providing a brief overview of some efficacy data from recent clinical studies with DPP-IV inhibitors. PMID- 16375747 TI - Design of cathepsin K inhibitors for osteoporosis. AB - Osteoporosis is a progressive, debilitating bone disease resulting in increased cost and morbidity to the elderly. This review summarizes the therapeutic approaches taken in the treatment of osteoporosis with particular emphasis on cathepsin K inhibitors. Cathepsin K, a cysteine protease predominantly expressed in osteoclasts, is a key player involved in bone matrix degradation. Both genetic ablation and small molecule inhibitor strategies versus cathepsin K have validated the importance of this enzyme in bone resorption. Starting from aldehyde-based leads, this review synopsizes the design of improved small molecule inhibitors by GlaxoWellcome researchers. These efforts involved the evaluation of various warheads, including cyanamides, ketoheterocycles, and ketoamides. Initial structure/activity relationships of aldehyde-based inhibitors proved useful in the design of ketoamide-based cathepsin K inhibitors. Further exploration of S(3), S(2), S(1), and S(1') subsites with P(3), P(2), P(1), and P(1') probes have resulted in the identification of potent, selective, orally bioavailable ketoamide-based inhibitors of cathepsin K with demonstrated in vivo efficacy. PMID- 16375748 TI - Small molecule coagulation cascade inhibitors in the clinic. AB - Venous thromboembolic disease, including deep vein thrombosis and pulmonary embolism, is a cause of significant mortality and morbidity. For several decades, anticoagulant options for the treatment and prevention of thrombosis have been limited mainly to agents such as unfractionated heparin and oral vitamin K antagonists such as warfarin. Although these therapies have proven benefits, they also have important limitations that result in their underuse in routine clinical practice. A variety of novel anticoagulants with improved pharmacologic and clinical profiles are in development, offering benefits over traditional therapies. Specifically, progress has been made in the development of small molecule Factor Xa inhibitors and thrombin inhibitors. The most advanced drugs reviewed include DPC-423, DPC-602, razaxaban, GSK's 813893, Portola's Xa inhibitors (formerly Millennium), otamixaban, DU-176b, KFA-1982, BAY-59-7939, DX 9065a, YM-150, LY-517717, Exanta, 3DP's thrombin inhibitors, SSR-182289, LB 30057, LB-30870, BIBR-1048 and Merck's thrombin inhibitors. With their potentially consistent and predictable pharmacological profile, oral formulation, and decreased need for coagulation monitoring, these new agents will likely increase the use and duration of anticoagulation treatment in thromboembolic disorders and reduce the burden associated with long-term management. PMID- 16375749 TI - Caspase inhibitors: a pharmaceutical industry perspective. AB - Caspase inhibition has been demonstrated to be therapeutically effective in moderating excessive programmed cell death, or apoptosis. Publications detailing programs in the pharmaceutical industry have been more frequent in recent years, ranging from SAR studies to clinically relevant animal models of disease. A summary of the work published in this exciting new area is presented, outlining the broad applicability of this fundamental cellular mechanism across several disease indications. This area of research has matured to the level of advancing compounds into clinical trials: VX-740 (Pralnacasan) and VX-765 as anti inflammatory agents, and IDN-6556, a pan-caspase inhibitor as an anti-apoptotic agent. PMID- 16375751 TI - A2B adenosine receptor antagonists: recent developments. AB - There are pharmacological evidences that A(2B) receptors are involved in inflammatory processes, such as asthma. For this reason, many efforts has been made for identifying selective A(2B) antagonists as anti-asthmatic agents. The updated material related to this field has been rationalised and arranged in order to offer an overview of the topic. PMID- 16375752 TI - Role of 1,4-benzothiazine derivatives in medicinal chemistry. AB - 1,4-Benzothiazine (1,4-BT) derivatives have been reported to exhibit a wide range of pharmacological properties including antifungal, immunostimulating, anti aldoso-reductase, anti-rheumatic, anti-allergic, vasorelaxant, anti-arrhythmic, anti-hypertensive, neuroprotective and cytotoxic activities. These different effects indicate that 1,4-BT is a template potentially useful in medicinal chemistry research and therapeutic applications. PMID- 16375753 TI - A re-emerging class of antimicrobial agents: streptogramins (quinupristin/dalfopristin) in the management of multiresistant gram-positive nosocomial cocci in hospital setting. AB - Multiresistant gram-positive cocci, including Staphylococcus aureus, coagulase negative staphylococci, Enterococcus faecalis and Enterococcus faecium, are emerging pathogens in the setting of immunocompromised, hospitalized patients, especially when surgery or invasive procedures are of concern, and patients are admitted in intensive care units. The spectrum of antimicrobial compounds available for an effective treatment of these infection is significantly threatened by the emerging and spread of glycopeptide-resistant strains. Quinupristin/dalfopristin is a novel streptogramine association, which represents an effective response to most of these problems, due to its innovative mechanim of action, its maintained activity against multiresistant pathogens, and its possibility of synergistic activity with other compounds. Problems related to the epidemiology of multiresistant gram-positive infection, potential clinical indications of quinupristin/dalfopristin, and updated data on efficacy and tolerability of this compound and its derivatives, are outlined on the ground of a review of available literature evidences. PMID- 16375754 TI - QT interval prolongation: and the beat goes on. AB - Consideration of QT interval prolongation and the risk for developing torsade de pointes is a critical issue in the evaluation of new bioactive agents. Over the past several years, there has been a dramatic increase in understanding the I(Kr) channel and its role in the duration of the action potential and cardiac repolarization. Furthermore, a variety of factors and situations have been identified that can increase the risk of QT interval prolongation. In this brief summary, an overview of the hERG channel and QT prolongation will be presented. The basic electro-physiology of the heart, the related action potentials, and pre clinical assays is reviewed. Further, an introduction to the current status of in silico efforts in predicting potential hERG blockers is discussed. Lastly, the strengths and weaknesses of each modeling method is presented along with insight to the appropriate use of each model. PMID- 16375755 TI - Modulation of HIV-1 transcription by cytokines and chemokines. AB - Infection with HIV results in the modulation of circulating levels of many host factors. Several host proteins that are up-regulated in HIV infection have the potential to influence virus replication. More specifically, the transcription of HIV-1 can be modulated in vivo by host proteins, including cytokines and chemokines. Cytokines modulate transcription mediated by the HIV-1 long terminal repeat (LTR) via multiple signal transduction pathways with resulting recruitment of numerous transcription factors, including NFkappaB, C/EBP, AP-1, TCF-1alpha, NF-IL-6 and ISGF-3. The effects on transcription may vary depending upon the cell type studied and upon the timing of the exposure of infected or transfected cells to cytokines. Furthermore, studies of cytokine mediated activation or inhibition of LTR mediated transcription may also be affected by the presence of the HIV-1 trans-activating protein, Tat, which has significant impact upon the redox state of the cell. This review will examine the complexities of the positive and negative control of HIV transcription by cytokines and chemokines. PMID- 16375756 TI - Recent advances in discovery and development of promising therapeutics against hepatitis C virus NS5B RNA-dependent RNA polymerase. AB - Lack of highly effective and safe therapeutics for hepatitis C virus (HCV) infection provides an opportunity as well as a challenge to discover novel and potent anti-HCV drugs. HCV NS5B RNA-dependent RNA polymerase (RdRp) is responsible for viral genome replication and thus constitutes a valid target for therapeutic intervention. To date, numerous HCV NS5B RdRp inhibitors have been discovered. This review focuses on the recent advances in discovery, mechanism of action studies and biological characterization of several distinct classes of potent inhibitors for NS5B RdRp. The clinical efficacy and developmental status of several promising compounds are also outlined. PMID- 16375757 TI - Thymidine phosphorylase inhibitors: recent developments and potential therapeutic applications. AB - Thymidine Phosphorylase (TPase) catalyses the reversible phosphorolysis of pyrimidine 2'-deoxynucleosides to 2-deoxyribose-1-phosphate and their respective pyrimidine bases, including the phosphorolysis of nucleoside analogues with important antiviral or anticancer properties. Moreover, TPase, identified also as the angiogenic platelet-derived endothelial cell growth factor (PD-ECGF), stimulates endothelial cell migration in vitro and angiogenesis in vivo and plays an important role in tumour progression and metastasis. Here we have summarized the most recent approaches in the search for novel TPase inhibitors together with the potential therapeutic applications of such inhibitors. PMID- 16375758 TI - Involvement of PI3K/Akt pathway in prostate cancer--potential strategies for developing targeted therapies. AB - This review presents some therapeutic interventions actually considered in prostate cancer therapy to compensate constitutive activation of the PI3K/Akt signalling pathway induced, particularly, by mutations of PTEN gene. Special emphasis is placed on applicability of EGF-R tyrosine kinase, COX-2, PDK-1, mTOR and farnesyltransferase inhibitors. PMID- 16375759 TI - Synthetic approaches to the 2004 new drugs. AB - New drugs are introduced to the market every year and each individual drug represents a privileged structure for its biological target. In addition, these new chemical entities (NCEs) not only provide insights into molecular recognition, but also serve as leads for designing future drugs. To this end, this review covers the syntheses of 12 NCEs marketed in 2004. PMID- 16375760 TI - Protocol for Project FACT: a randomised controlled trial on the effect of a walking program and vitamin B supplementation on the rate of cognitive decline and psychosocial wellbeing in older adults with mild cognitive impairment [ISRCTN19227688]. AB - BACKGROUND: The prevalence of individuals with cognitive decline is increasing since the number of elderly adults is growing considerably. The literature provides promising results on the beneficial effect of exercise and vitamin supplementation on cognitive function both in cognitively healthy as well as in the demented elderly. METHODS/DESIGN: The design is a two-by-two factorial randomised controlled trial. The study population consists of independently living elderly, between 70 and 80 years old, with mild cognitive impairment (MCI). In the RCT the effect of two interventions, a walking program and vitamin supplementation, is examined. The walking program (WP) is a group-based program aimed at improving cardiovascular endurance; frequency two lessons a week; lesson duration one hour; program duration one year. Non-walking groups receive a placebo activity program (PAP) (i.e. low intensive non-aerobic group exercises, like stretching) with the same frequency, lesson and program duration. Vitamin supplementation consists of a single daily vitamin supplement containing 50 mg B6, 5 mg folic acid and 0,4 mg B12 for one year. Subjects not receiving vitamin supplements are daily taking an identically looking placebo pill, also for a year. Participants are randomised to four groups 1) WP and vitamin supplements; 2) WP and placebo supplements; 3) PAP and vitamin supplements; 4) PAP and placebo supplements. Primary outcome measures are measures of cognitive function. Secondary outcomes include psychosocial wellbeing, physical activity, cardiovascular endurance and blood vitamin levels. DISCUSSION: No large intervention study has been conducted yet on the effect of physical activity and vitamin supplementation in a population-based sample of adults with MCI. The objective of the present article is to describe the design of a randomised controlled trial examining the effect of a walking program and vitamin B supplementation on the rate of cognitive decline in older adults with MCI. PMID- 16375761 TI - An indirect enzyme-linked immunosorbent assay for rapid and quantitative assessment of Type III virulence phenotypes of Pseudomonas aeruginosa isolates. AB - BACKGROUND: The presence of a Type III secretion system in clinical isolates of Pseudomonas aeruginosa is associated with severe disease and poor outcomes in infections caused by this pathogen. We describe an indirect enzyme-linked immunosorbent assay that rapidly and quantitatively detects two exotoxins, ExoU and ExoT, and two structural components, PopD and PcrV, of the P. aeruginosa Type III secretion system after in-vitro growth in a calcium-free minimal medium. METHODS: We used this assay to characterize the Type III secretion phenotype of 74 clinical isolates of P. aeruginosa. Findings were compared with results of standard immunoblotting and correlated with Type III secretion-dependent virulence of isolates toward cultured epithelial cells. RESULTS: Results of the ELISA assay were concordant with immunoblot detection of the secreted antigens for 73 of 74 isolates. The Type III secretion phenotype assessed by this immunoassay predicted bacterial virulence toward epithelial cells in vitro for all but five of the clinical isolates. CONCLUSION: The availability of an ELISA assay for rapid detection of Type III secreted virulence factors will facilitate large clinical studies to examine whether the Type III secretion phenotype of a P. aeruginosa isolate predicts the course of clinical disease in a patient and should be taken into account in determining optimal treatment strategies for infected patients. PMID- 16375762 TI - Insulin allergy and resistance successfully treated by desensitisation with Aspart insulin. AB - A 25-year-old, with type I Diabetes Mellitus with a previous diagnosis of Protamine Allergy but not to human Insulin, started to notice anaphylactic reactions immediately after bolus with Insulin. Skin prick and intradermal test were positive to all insulins. Skin tests to other potential allergens resulted negative. Examination after bolus of Human Insulin revealed urticaria. Daily insulin requirement were around 2-2,4 U/Kg/day. Slow desensitisation with Aspart insulin, the insulin with lowest size of skin test, was performed using subcutaneous insulin pump. Six months after the end of desensitisation his daily insulin requirement decreased to 0.8 U/Kg/day and oral corticosteroids are being reduced with no symptoms. PMID- 16375764 TI - Relationship of 99mtechnetium labelled macroaggregated albumin (99mTc-MAA) uptake by colorectal liver metastases to response following Selective Internal Radiation Therapy (SIRT). AB - BACKGROUND: SIRT is an emerging treatment for liver tumours which relies on the selective uptake by tumour of 90Y microspheres following hepatic arterial injection. Response rates of around 90% are reported. Hepatic arterial injection of MAA gives an indication of the expected distribution of 90Y microspheres within the liver. This study sought to determine if the MAA scan could be predictive of subsequent tumour response. METHODS: 58 patients with colorectal hepatic metastases received SIRT. All had pre-treatment MAA planar images and CT scans which were retrospectively reviewed. Tumours were qualitatively considered "cold", "equivocal" or "hot" based on MAA uptake and the ratio of uptake in tumour and normal liver tissue was calculated (TNR). Following SIRT (which included the administration of hepatic arterial Angiotensin 2) tumour response was assessed by CEA changes one to two months after treatment and by serial CT. RESULTS: Uptake was classified as "hot" in 37 patients (Group 1) and "equivocal" or "cold" in 21 (Group 2). CEA levels fell dramatically in over 90% of patients. The falls were not significantly different between the groups. There was no correlation between TNR and tumour response based on CEA changes (r2 = 0.004). CT responses after 3 months were not different in the 2 Groups. CONCLUSION: The pattern of MAA uptake by colorectal liver tumours after arterial injection is not a predictor of tumour response after treatment by SIRT. The results suggest the doses of 90Y microspheres used may be greater than is necessary. PMID- 16375763 TI - Evolutionary programming as a platform for in silico metabolic engineering. AB - BACKGROUND: Through genetic engineering it is possible to introduce targeted genetic changes and hereby engineer the metabolism of microbial cells with the objective to obtain desirable phenotypes. However, owing to the complexity of metabolic networks, both in terms of structure and regulation, it is often difficult to predict the effects of genetic modifications on the resulting phenotype. Recently genome-scale metabolic models have been compiled for several different microorganisms where structural and stoichiometric complexity is inherently accounted for. New algorithms are being developed by using genome scale metabolic models that enable identification of gene knockout strategies for obtaining improved phenotypes. However, the problem of finding optimal gene deletion strategy is combinatorial and consequently the computational time increases exponentially with the size of the problem, and it is therefore interesting to develop new faster algorithms. RESULTS: In this study we report an evolutionary programming based method to rapidly identify gene deletion strategies for optimization of a desired phenotypic objective function. We illustrate the proposed method for two important design parameters in industrial fermentations, one linear and other non-linear, by using a genome-scale model of the yeast Saccharomyces cerevisiae. Potential metabolic engineering targets for improved production of succinic acid, glycerol and vanillin are identified and underlying flux changes for the predicted mutants are discussed. CONCLUSION: We show that evolutionary programming enables solving large gene knockout problems in relatively short computational time. The proposed algorithm also allows the optimization of non-linear objective functions or incorporation of non-linear constraints and additionally provides a family of close to optimal solutions. The identified metabolic engineering strategies suggest that non-intuitive genetic modifications span several different pathways and may be necessary for solving challenging metabolic engineering problems. PMID- 16375765 TI - Discontinuation of treatment of schizophrenic patients is driven by poor symptom response: a pooled post-hoc analysis of four atypical antipsychotic drugs. AB - BACKGROUND: Stopping antipsychotic treatment can interrupt improvement and exacerbate the illness. The reasons for discontinuing treatment during controlled clinical trials were analyzed to explore this phenomenon. METHODS: A post-hoc, pooled analysis was made of 4 randomized, double-blind clinical trials, 24-28 weeks in duration, involving 1627 patients with schizophrenia or a related disorder. Analyses combined all the atypical antipsychotic treatment groups in the studies. RESULTS: The majority of patients (53%) stopped their treatment at an early stage. Poor psychiatric response along with worsening symptoms was the most frequently given reason for discontinuing the course (36%), which was substantially more common than discontinuation due to poor tolerability of the medication (12%). This phenomenon was corroborated by less improvement in patients who discontinued treatment compared with those who completed, based on the PANSS total scores. Discontinuation due to poor response was, apparently, more predominantly linked to patient perception than to physicians' conclusions alone (80% vs. 20%). Discontinuation due to patient perception of poor response appeared to occur particularly early in the course of treatment. Patients who discontinued due to poor toleration of the medication responded in a more comparable manner with completers. CONCLUSION: Discontinuing treatment may lead to exacerbation of symptoms, undermining therapeutic progress. In these studies, poor response to treatment and worsening of underlying psychiatric symptoms, and to a lesser extent, intolerability to medication were the primary contributors to treatment being discontinued. Our findings suggest that adherence may be enhanced by effective symptom control, as objectively measured and as subjectively perceived. Such strategies may improve patients' willingness to undertake long term therapy and increase the likelihood of a better prognosis. PMID- 16375766 TI - SW-620 cells treated with topoisomerase I inhibitor SN-38: gene expression profiling. AB - BACKGROUND: The goal of this study was to evaluate changes in gene expression in SW-620 cells in response to SN-38 in order to further elucidate the mechanisms by which SN-38 causes apoptosis and cell cycle arrest. METHODS: We used a quantitative gene expression microarray assay to identify the genes regulated by SN-38 treatment in colon cancer cells and confirmed our results with RT-PCR. By gene expression profiling, we first screened a proprietary list of about 22,000 genes. RESULTS: Treatment with SN-38 cells resulted in two-fold or greater alteration in the level of expression of 192 genes compared to control treatment. Most of the affected genes were not known to be responsive to SN-38 prior to this study. SN-38 treatment of these cells was found to affect the expression of various genes involved in DNA replication, transcription, signal transduction, growth factors, cell cycle regulation, and apoptosis, as well as other genes with unknown function. Changes in expression of 14 genes were confirmed by quantitative real-time polymerase chain reaction (RT-PCR). CONCLUSION: This study leads to an increased understanding of the biochemical pathways involved in SN-38 induced apoptosis and possibly to the identification of new therapeutic targets. PMID- 16375767 TI - The diagnostic value of biomarkers (SteatoTest) for the prediction of liver steatosis. AB - BACKGROUND: Biopsy is the usual gold standard for liver steatosis assessment. The aim of this study was to identify a panel of biomarkers (SteatoTest), with sufficient predictive values, for the non-invasive diagnosis of steatosis in patients with or without chronic liver disease. Biomarkers and panels were assessed in a training group of consecutive patients with chronic hepatitis C and B, alcoholic liver disease, and non-alcoholic fatty liver disease, and were validated in two independent groups including a prospective one. Steatosis was blindly assessed by using a previously validated scoring system. RESULTS: 310 patients were included in the training group; 434 in three validation groups; and 140 in a control group. SteatoTest was constructed using a combination of the 6 components of FibroTest-ActiTest plus body mass index, serum cholesterol, triglycerides, and glucose adjusted for age and gender. SteatoTest area under the ROC curves was 0.79 (SE = 0.03) in the training group; 0.80 (0.04) in validation group 1; 0.86 (0.03) in validation group 2; and 0.72 (0.05) in the validation group 3 - all significantly higher than the standard markers: gamma-glutamyl transpeptidase or alanine aminotransferase. The median SteatoTest value was 0.13 in fasting controls; 0.16 in non-fasting controls; 0.31 in patients without steatosis; 0.39 in grade 1 steatosis (0-5%); 0.58 in grade 2 (6-32%); and 0.74 in grade 3-4 (33-100%). For the diagnosis of grade 2-4 steatosis, the sensitivity of SteatoTest at the 0.30 cut-off was 0.91, 0.98, 1.00 and 0.85 and the specificity at the 0.70 cut-off was 0.89, 0.83, 0.92, 1.00, for the training and three validation groups, respectively. CONCLUSION: SteatoTest is a simple and non invasive quantitative estimate of liver steatosis and may reduce the need for liver biopsy, particularly in patients with metabolic risk factor. PMID- 16375768 TI - Dynamic 3D echocardiography in virtual reality. AB - BACKGROUND: This pilot study was performed to evaluate whether virtual reality is applicable for three-dimensional echocardiography and if three-dimensional echocardiographic 'holograms' have the potential to become a clinically useful tool. METHODS: Three-dimensional echocardiographic data sets from 2 normal subjects and from 4 patients with a mitral valve pathological condition were included in the study. The three-dimensional data sets were acquired with the Philips Sonos 7500 echo-system and transferred to the BARCO (Barco N.V., Kortrijk, Belgium) I-space. Ten independent observers assessed the 6 three dimensional data sets with and without mitral valve pathology. After 10 minutes' instruction in the I-Space, all of the observers could use the virtual pointer that is necessary to create cut planes in the hologram. RESULTS: The 10 independent observers correctly assessed the normal and pathological mitral valve in the holograms (analysis time approximately 10 minutes). CONCLUSION: this report shows that dynamic holographic imaging of three-dimensional echocardiographic data is feasible. However, the applicability and use-fullness of this technology in clinical practice is still limited. PMID- 16375769 TI - Cloxacillin versus vancomycin for presumed late-onset sepsis in the Neonatal Intensive Care Unit and the impact upon outcome of coagulase negative staphylococcal bacteremia: a retrospective cohort study. AB - BACKGROUND: Coagulase negative staphylococcus (CONS) is the main cause of late onset sepsis in Neonatal Intensive Care Units (NICU). Although CONS rarely causes fulminant sepsis, vancomycin is frequently used as empiric therapy. Indiscriminate use of vancomycin has been linked to the emergence of vancomycin resistant organisms. The objective of this study was to compare duration of CONS sepsis and mortality before and after implementation of a policy of selective vancomycin use and compare use of vancomycin between the 2 time periods. METHODS: A retrospective study was conducted of infants > or =4 days old, experiencing signs of sepsis with a first positive blood culture for CONS, during two 12-month periods. Late-onset sepsis was treated empirically with vancomycin and gentamicin during period 1, and cloxacillin and gentamicin during period 2. The confidence interval method was used to assess non-inferiority of the outcomes between the two study groups. RESULTS: There were 45 episodes of CONS sepsis during period 1 and 37 during period 2. Duration of sepsis was similar between periods (hazard ratio of 1.00, 95%CI: 0.64, 1.57). One death during period 2 was possibly related to CONS sepsis versus none in period 1. Vancomycin was used in 97.8% of episodes in period 1 versus 81.1% of episodes in period 2. CONCLUSION: Although we failed to show non-inferiority of duration of sepsis in the cloxacillin and gentamicin group compared to the vancomycin and gentamicin group, duration of sepsis was clinically similar. Restricting vancomycin for confirmed cases of CONS sepsis resistant to oxacillin appears effective and safe, and significantly reduces vancomycin use in the NICU. PMID- 16375770 TI - Neurogenetic interactions and aberrant behavioral co-morbidity of attention deficit hyperactivity disorder (ADHD): dispelling myths. AB - BACKGROUND: Attention Deficit Hyperactivity Disorder, commonly referred to as ADHD, is a common, complex, predominately genetic but highly treatable disorder, which in its more severe form has such a profound effect on brain function that every aspect of the life of an affected individual may be permanently compromised. Despite the broad base of scientific investigation over the past 50 years supporting this statement, there are still many misconceptions about ADHD. These include believing the disorder does not exist, that all children have symptoms of ADHD, that if it does exist it is grossly over-diagnosed and over treated, and that the treatment is dangerous and leads to a propensity to drug addiction. Since most misconceptions contain elements of truth, where does the reality lie? RESULTS: We have reviewed the literature to evaluate some of the claims and counter-claims. The evidence suggests that ADHD is primarily a polygenic disorder involving at least 50 genes, including those encoding enzymes of neurotransmitter metabolism, neurotransmitter transporters and receptors. Because of its polygenic nature, ADHD is often accompanied by other behavioral abnormalities. It is present in adults as well as children, but in itself it does not necessarily impair function in adult life; associated disorders, however, may do so. A range of treatment options is reviewed and the mechanisms responsible for the efficacy of standard drug treatments are considered. CONCLUSION: The genes so far implicated in ADHD account for only part of the total picture. Identification of the remaining genes and characterization of their interactions is likely to establish ADHD firmly as a biological disorder and to lead to better methods of diagnosis and treatment. PMID- 16375771 TI - HIV prevalence among aboriginal British Columbians. AB - CONTEXT: There is considerable concern about the spread of HIV disease among Aboriginal peoples in British Columbia. OBJECTIVE: To estimate the number of Aboriginal British Columbians infected with HIV. DESIGN AND SETTING: A population based analysis of Aboriginal men and women in British Columbia, Canada from 1980 to 2001. PARTICIPANTS: Epidemic curves were fit for gay and bisexual men, injection drug users, men and women aged 15 to 49 years and persons over 50 years of age. MAIN OUTCOME MEASURES: HIV prevalence for the total Aboriginal population was modeled using the UNAIDS/WHO Estimation and Projection Package (EPP). Monte Carlo simulation was used to estimate potential number infected for select transmission group in 2001. RESULTS: A total of 170,025 Aboriginals resided in British Columbia in 2001, of whom 69% were 15 years and older. Of these 1,691 (range 1,479-1,955) men and women aged 15 years and over were living with HIV with overall prevalence ranging from 1.26% to 1.66%. The majority of the persons infected were men. Injection drug users (range 1,202-1,744) and gay and bisexual men (range 145, 232) contributed the greatest number of infections. Few persons infected were from low risk populations. CONCLUSION: More than 1 in every 100 Aboriginals aged 15 years and over was living with HIV in 2001. Culturally appropriate approaches are needed to tailor effective HIV interventions to this community. PMID- 16375772 TI - Parkin-deficient mice are not more sensitive to 6-hydroxydopamine or methamphetamine neurotoxicity. AB - BACKGROUND: Autosomal recessive juvenile parkinsonism (AR-JP) is caused by mutations in the parkin gene which encodes an E3 ubiquitin-protein ligase. Parkin is thought to be critical for protecting dopaminergic neurons from toxic insults by targeting misfolded or oxidatively damaged proteins for proteasomal degradation. Surprisingly, mice with targeted deletions of parkin do not recapitulate robust behavioral or pathological signs of parkinsonism. Since Parkin is thought to protect against neurotoxic insults, we hypothesized that the reason Parkin-deficient mice do not develop parkinsonism is because they are not exposed to appropriate environmental triggers. To test this possibility, we challenged Parkin-deficient mice with neurotoxic regimens of either methamphetamine (METH) or 6-hydroxydopamine (6-OHDA). Because Parkin function has been linked to many of the pathways involved in METH and 6-OHDA toxicity, we predicted that Parkin-deficient mice would be more sensitive to the neurotoxic effects of these agents. RESULTS: We found no signs consistent with oxidative stress, ubiquitin dysfunction, or degeneration of striatal dopamine neuron terminals in aged Parkin-deficient mice. Moreover, results from behavioral, neurochemical, and immunoblot analyses indicate that Parkin-deficient mice are not more sensitive to dopaminergic neurotoxicity following treatment with METH or 6-OHDA. CONCLUSION: Our results suggest that the absence of a robust parkinsonian phenotype in Parkin-deficient mice is not due to the lack of exposure to environmental triggers with mechanisms of action similar to METH or 6-OHDA. Nevertheless, Parkin-deficient mice could be more sensitive to other neurotoxins, such as rotenone or MPTP, which have different mechanisms of action; therefore, identifying conditions that precipitate parkinsonism specifically in Parkin deficient mice would increase the utility of this model and could provide insight into the mechanism of AR-JP. Alternatively, it remains possible that the absence of parkinsonism in Parkin-deficient mice could reflect fundamental differences between the function of human and mouse Parkin, or the existence of a redundant E3 ubiquitin-protein ligase in mouse that is not found in humans. Therefore, additional studies are necessary to understand why Parkin-deficient mice do not display robust signs of parkinsonism. PMID- 16375773 TI - Cystathionine beta-synthase T833C/844INS68 polymorphism: a family-based study on mentally retarded children. AB - BACKGROUND: Cystathionine beta-synthase (CBS) mediates conversion of homocysteine to cystathionine and deficiency in enzyme activity may lead to hyperhomocysteinemia/homocystinuria, which are often associated with mental retardation (MR). A large number of polymorphisms have been reported in the CBS gene, some of which impair its activity and among these, a T833C polymorphism in cis with a 68 bp insertion at 844 in the exon 8 is found to be associated with mild hyperhomocysteinemia in different ethnic groups. METHODS: The present study is aimed at investigating the association between T833C/844ins68 polymorphism and MR. One hundred and ninety MR cases were recruited after psychometric evaluation. Hundred and thirty-eight control subjects, two hundred and sixty-seven parents of MR probands and thirty cardiovascular disorder (CVD) patients were included for comparison. Peripheral blood was collected after obtaining informed written consent. The T833C/844ins68 polymorphism was investigated by PCR amplification of genomic DNA and restriction fragment length polymorphism analysis, followed by statistical analysis. RESULTS: The genotypic distribution of the polymorphism was within the Hardy-Weinberg equilibrium. A slightly increased genotypic frequency was observed in the Indian control population as compared to other Asian populations. Both haplotype-based haplotype relative risk analysis and transmission disequilibrium test reveled lack of association of the T833C/844ins68 polymorphism with MR; nevertheless, the relative risk calculated was higher (>1) and in a limited number of informative MR families, preferential transmission of the double mutant from heterozygous mothers to the MR probands was noticed (chi2 = 4.00, P < 0.05). CONCLUSION: This is the first molecular genetic study of CBS gene dealing with T833C/844ins68 double mutation in MR subjects. Our preliminary data indicate lack of association between T833C/844ins68 polymorphism with MR. However, higher relative risk and biased transmission of the double mutation from heterozygous mothers to MR probands are indicative of a risk of association between this polymorphism with mental retardation. PMID- 16375774 TI - Maintaining ethical standards in medical publishing. PMID- 16375775 TI - The future of British surgery. PMID- 16375776 TI - A comparative evaluation of ear diseases in children of higher versus lower socioeconomic status. AB - This study was conducted with an aim to assess the prevalence and profile of ear diseases in children from the higher and lower socioeconomic strata of society. Two groups of schools within Delhi were selected. Group A comprised of government schools located in slum areas with an average parental income of INR (Indian National Rupees) 1050 per month and group B of elite private schools with an average family income of TNR 35,000 per month. Three thousand children between the ages of 5-12 years were screened with the help of a written proforma and ear examination. Tympanometry and audiometry were done, where required. 19.6 per cent of children of group A were found to be suffering from ear diseases compared to 2.13 per cent of group B children. The two groups were also compared for number of family members, status of hygiene and parental education. PMID- 16375777 TI - Is phenol a safe local anaesthetic for grommet insertion? AB - Two studies were performed to determine whether topical phenol is a safe and well tolerated local anaesthetic for grommet insertion. Study 1 was a retrospective examination of audiological outcomes and complications. Data were obtained regarding 71 procedures in 57 patients. One late infection and nine early extrusions were noted. No statistically significant changes between pre- and post operative bone conduction thresholds were found. Study 2 was a prospective analysis of patients' perceptions of the procedure. Data from 17 patient questionnaires were analysed as follows: pain rating--not painful, three patients; slightly painful, 14 patients. Overall experience rating--pleasant, four patients; slightly unpleasant, 10; unpleasant, three. All patients stated that they would undergo the procedure again. In conclusion, we found no evidence of phenol-induced hearing loss. The complication rate was within normal limits and patients were satisfied with the procedure. Grommet insertion using phenol as a local anaesthetic is safe and acceptable to patients. PMID- 16375778 TI - Takayasu's arteritis and saddle nose deformity: a new association. AB - Aortitis and saddle nose deformity are extremely unusual manifestations of a variety of systemic diseases. The concurrent appearance of these apparently disparate clinical features is a clinical rarity. A case of saddle nose deformity in a patient with confirmed Takayasu's arteritis is presented. The relevant literature is reviewed with reference to the possible differential diagnosis of patients with aortitis and saddle nose deformity. PMID- 16375779 TI - A novel application of the adult Merocel nasal pack. PMID- 16375781 TI - Delivery of child development services by videoconferencing: a review of four years' experience in Queensland. AB - In 2001 the Child Development Unit (CDU) in Brisbane piloted a series of monthly multidisciplinary case discussions via videoconference in the area of child development. During 2001 two sessions were provided; during 2004 there were 40. The substantial growth in 2004 was due to the expansion of child development services to include special interest group meetings and multipoint case conference meetings. In 2004, a total of 49 h of videoconferencing was conducted. The average session length was 75 min. Education and training sessions were delivered to 32 hospitals and health centres in Queensland and northern New South Wales. The maximum number of sites involved during a single videoconference was 25. The average number of attendees for each videoconference was five per site, including allied health staff, nurses and paediatricians. The delivery of child development services via videoconference has been shown to be useful in Queensland, especially for allied health staff working in regional and remote areas. The growth of the programme indicates its acceptance as a mainstream child development service in Queensland. PMID- 16375782 TI - The iPath telemedicine platform. AB - An early, point-to-point telepathology system at the University of Basel developed into an open-source, Internet-based platform for telemedicine in 2001. The Internet Pathology Suite (iPath) is a Web-based telepathology platform that permits the online presentation and discussion of cases within user groups. It also allows realtime telemicroscopy across firewalls. After four years, the telepathology network has over 700 active users. More than 6,300 cases with a total of about 39,000 images have been diagnosed. The diagnostic workload of all these cases is not exclusively handled by the Department of Pathology in Basel, but by a growing number of independent groups who also use the server simply as a case repository. What started as a small project for hospitals in Switzerland has become a global network. PMID- 16375783 TI - The Telemedicine Information Exchange: 10 years' experience. AB - The Telemedicine Information Exchange (TIE) Website has provided information on telemedicine since 1995. The site could be considered successful from its longevity alone; however, it can also be considered a success using other measures. The usage of the site has grown steadily over the years, to more than 20,000 visitors per month by 2005. The Bibliographic database has over 16,000 telemedicine-related citations, more than found anywhere else, and is the most visited section of the Website. The second most visited database, Active Telemedicine Programs, is the result of 10 years of tracking activity and growth using a voluntary online survey. This continuous monitoring of telemedicine literature and activity has allowed TIE research staff to anticipate user needs and interests. For instance, a Home Telehealth section was initiated after it became apparent that interest in the subject was increasing. Longevity also provides a perspective about the best ways of maintaining a content-heavy online resource. Efforts to augment federal funding (the TIE's sole source of support) by selling packaged information products have taught us that online users will not pay for information. Our raison d'etre for the past 10 years has been the provision of credible, up-to-date information, and our reward has been positive feedback from thousands of TIE users. PMID- 16375784 TI - Success and failure in Web-based medical collaboration. AB - A Web-based collaboration tool was developed for ophthalmologists, called Oyenet. A feasibility study was performed in 2000. The system was ready to use in 2001, but the usage of the system was disappointing, compared to the enthusiasm expressed in the feasibility study. In order to identify facilitators and barriers for usage of the system, we performed a qualitative study. Nineteen ophthalmologists from different parts of Norway, who had been using the system but to a varying degree, were interviewed. The study revealed that a single common solution is not appropriate for the ophthalmology community as a whole. It also revealed that colleagues from abroad should be allowed to use the system, in order for it to act as a tool for collaboration and seeking advice in complex medical problem solving. PMID- 16375785 TI - Telemedical treatment at home of diabetic foot ulcers. AB - The treatment of diabetic foot ulcers may not always be well organized and not all patients have access to expert evaluation. We investigated the use of telemedicine to enable a visiting nurse (in the patient's home) to coordinate the treatment with experts (at the hospital). The equipment consisted of a UMTS videophone and an Internet-based patient record. Field studies were carried out at the outpatient clinic and by following visiting nurses. A total of 15 participants were invited to five workshops and experiments held in our laboratories, at the hospital, and between hospital and patients' homes. Finally, in a pilot test, five patients were offered three teleconsultations at their homes in lieu of visits to the hospital. Preliminary results were promising: (1) both clinicians and patients found the equipment easy to use; (2) the doctor could prescribe treatment at a distance; (3) the visiting nurse had realtime contact with the hospital and treatment could begin immediately according to the doctor's orders; (4) the patient saved time in not having to travel to the hospital. PMID- 16375786 TI - Development of a pilot telemedicine network for paediatric oncology in Brazil. AB - We established a pilot telemedicine network for paediatric oncology in Brazil, linking the School of Medicine at the University of Sao Paulo in Sao Paulo City to the 'Hospital de Base' in Porto Velho, Rondonia, located in the Amazon region, 3,000 km away. The videoconferencing link used ISDN transmission at 384 kbit/s. The network was used for patient screening, follow-up, treatment monitoring and other activities. Between March 2000 and 2002, 69 videoconferences were held for 33 patients, 29 with cancer. During this period, 16 patients required transfer, 18 patients died and 11 achieved cancer remission. The main cause for patient mortality (infection) was not one that could be addressed directly by telemedicine. Using the School of Medicine as a benchmark, the average mortality rate for paediatric cancer patients in the pilot was higher. However, it was lower than previous levels observed at Rondonia (62% versus 80%). PMID- 16375787 TI - Evaluation of the Western Australian Department of Health telehealth project. AB - In 1999, the Department of Health in Western Australia began a telehealth project, which finished in 2004. The 75 videoconferencing sites funded by the project were part of a total state-wide videoconference network of 104 sites. During the period from January 2002 to December 2003, a total of 3,266 consultations, case reviews and patient education sessions took place. Clinical use grew to 30% of all telehealth activity. Educational use was approximately 40% (1,416 sessions) and management use was about 30% (1,031 sessions). The average overhead cost per telehealth session across all regions and usage types was 192 Australian dollars. Meaningful comparisons of the results of the present study with other public health providers were difficult, because many of the available Websites on telehealth were out of date. Despite the successful use of telehealth to deliver clinical services in Western Australia, sustaining the effort in the post-project phase will present significant challenges. PMID- 16375788 TI - Attitude to telemedicine, and willingness to use it, in audiology patients. AB - We studied the willingness of patients to use telemedicine for ear- and hearing related appointments, and the factors that influenced their decision to participate in telemedicine. A survey was designed with questions about patient appointments, perceived advantages and barriers to telemedicine, and prior use of the Internet for health-related matters. A total of 116 patients in four audiology centres were surveyed from December 2004 to May 2005. There were 54 male and 62 female respondents; 46% of the participants were aged over 66 years. In all, 75% had not previously heard of telemedicine. The most common reasons for willingness to use telemedicine were to reduce the time waiting for an appointment and cost. The most common barrier to using telemedicine was a preference for face-to-face visits. Of those surveyed, 32% were willing to use telemedicine, 10% would sometimes be willing, 28% were unsure, and 30% were not willing. There was no relationship between willingness and age or gender, except that women over the age of 55 years were less willing. Patients who had previously heard of telemedicine and used the Internet for health-related matters, especially men, were more inclined to have a telemedicine appointment. PMID- 16375789 TI - The clinical champion role in the development of a successful telehealth wound care project for remote Australia. AB - The role of a clinical champion in a wound care project was examined in terms of an emancipatory processes framework. During the project the role changed significantly, as the needs of the project changed. In the early phase of the project, the clinical champion's role was that of team leader. During the middle phase of the project, the clinical champion's role changed to health services advocate and coach. During the final phase of the project, the clinical champion's role changed again, to that of salesperson and academic. Experience with the clinical issue being addressed by the new service, and clear motivation to complete the project, thereby seeing the new service established, allowed the clinical champion to motivate the team to overcome the difficulties in the change process. PMID- 16375790 TI - Striving for evidence in e-health evaluation: lessons from health technology assessment. AB - Evaluation is crucial to the integration of e-health applications into the health care system and their ultimate sustainability. However, e-health evaluation is often criticized for the poor quality of research design, the lack of common outcome indicators and the absence of an agreed theory. Health technology assessment (HTA) could offer a sound methodological basis for e-health evaluation. However, there have been major concerns about the applicability of the HTA approach to the evaluation of e-health initiatives. Evaluators -- and decision makers -- must accept that telehealth evaluation may serve different purposes for different stakeholders, and therefore concede that no single evaluation framework or methodology, even the randomized controlled trial, is totally objective. To address the complex environment of telehealth evaluation, a participatory strategy is useful, whereby stakeholders are involved in the study design and definition of evaluation questions at each phase. This will also build confidence between the evaluation team and the stakeholders, facilitating informed decision making through an integrated knowledge mobilization activity. PMID- 16375791 TI - Challenges for implementing wireless hand-held technology in health care: views from selected Queensland nurses. AB - Many health-care providers in Australia are exploring the use of wireless technology to improve service delivery. It appears that the solutions so far have been dictated by the hardware vendors and that the business case is yet to drive the implementations. A focus group discussion was facilitated with eight senior management staff involved in health care in Western Australia. This resulted in a set of challenges, which were used to invite opinions from nursing staff in Queensland. A total of 31 interviews were conducted. The analysis returned a set of 63 themes, which were grouped. These groupings reflected the challenges as lack of user-friendly applications, unreliable technology, substandard testing, shortage of staff, concerns for security, reliance on technology, existing problems, work schedule, training, outdated health policy, coverage of wireless links, confidentiality and lack of awareness. The interviews clearly indicated the need for training and awareness procedures. The present study provides some of the information necessary to realize an enterprise-wide implementation of wireless technology. PMID- 16375792 TI - Achievements and challenges on policies for allied health professionals who use telehealth in the Canadian Arctic. AB - We formulated policies and procedures for allied health professionals (AHPs) who provide services using telehealth in Nunavut, Canada's newest Arctic territory. These are a supplement to the clinical policies and procedures already established for Nunavut physicians and nurses. The services were in the areas of audiology, dietetics/nutrition, midwifery, occupational therapy, ophthalmic services, pharmacy, physiotherapy, psychology, respiratory therapy, social work and speech therapy. Documents specific to each of the services were developed, drawing on information from Government of Nunavut data, Nunavut healthcare providers and links made through the Internet. Topics included the scope and limitations of telehealth services, staff responsibilities, training and reporting, professional standards and cultural considerations. We also considered generic policies covering common issues such as jurisdiction, licensing and liability. The policies and procedures for AHPs will enhance and expand the successes already achieved with telehealth in Nunavut. The challenges are to balance the preferred approaches to service provision with the realities of health care and communications in an Arctic setting. PMID- 16375793 TI - The LifeShirt system for wireless patient monitoring in the operating room. AB - We examined the functionality and reliability of a wearable physiological monitoring system (LifeShirt) during normal daily activities and in a hospital operating room (OR) environment. The garment collects physiological data such as oxygen saturation, and stores them in a recorder from which it can be read afterwards. Ten normal subjects wore the shirt continuously for 8 h per day. Feedback from the testers was quite positive, although the collected data varied in quality. Ten hospital patients also wore the shirt during endoscopy. The data collected during the hospital stay were qualitatively adequate. Measuring respiratory function caused the biggest problems. The study showed that intelligent garment technology could be used in an OR environment for patient monitoring, albeit not in realtime. It may also be useful in home follow-up. PMID- 16375794 TI - Telemedicine by email in remote Cambodia. AB - An email-based telemedicine service was implemented in two remote village communities in Cambodia. Volunteer physicians at the Brigham and Women's Hospital, Massachusetts General Hospital and the Sihanouk Hospital Center of Hope in Phnom Penh provide monthly consultations to the local clinicians. Between February 2001 and May 2005, there were 469 teleconsultations. The 214 telemedicine cases involving new patients managed in the first 28 months were reviewed. The mean duration of the chief complaint at the initial patient visit was 37 months for the first six months and had dropped to eight months by the end of the study period. Of 63 adult patients surveyed, all were either satisfied (54%, n = 34) or very satisfied (46%, n = 29) with their experience in the telemedicine clinic. About 78% (n = 49) were willing to pay, on average, 0.63 US dollars for their visits. The introduction of basic point-of-care laboratory testing in November 2004 was associated with a reduction in patients requiring off-site referral for completion of laboratory testing (69% before to 35% afterwards, P < 0.001). The success of the pilot telemedicine programme confirms the value of email support for non-physician health-care workers in the developing world. PMID- 16375795 TI - Difficulties in moving routine medical checks from the specialist level to the general practitioner level. AB - We began a project to move routine medical checks for appropriate patients from the specialist level to the patient's normal general practitioner (GP). The GP's analysis and conclusions would be checked by the specialist, using electronic messaging. The idea for the project came from the top level of the regional health authority. Despite that, the project was closed down before pilot testing began. We used stakeholder theory as a post-project evaluation to analyse what happened and where it went wrong. A common mistake in project planning is to focus the planning effort on system tasks and not to pay attention to a well thought-out handling of the project's stakeholders. This was what happened in our project. Ideal objectives and good political intentions are not enough to implement a new e-health service. PMID- 16375796 TI - Establishment of the Brazilian telehealth network for paediatric oncology. AB - A telemedicine network has been established in Brazil to support distance medical practice in paediatric oncology. ONCONET comprises a national network of universities, research institutes and medical institutions. The system is Web based and hosted on a high-performance computing infrastructure based on clusters of computers. It is based on open-source software, designed to provide high performance, fault tolerance and high availability. The ONCONET began operation in 2004. Currently, 30 hospitals affiliated to the Brazilian Society for Paediatric Oncology are users of the ONCONET. Six hospitals are connected by broadband access through the National Education and Research Network and 24 by conventional Internet access. The Multimedia Patients Registry also became operational in 2004, and its database contains information on 3,200 patients from the 30 hospitals. The technological platform was notable for its low production cost. It thus appears to be a sustainable solution to the problem of delivering continuing medical education in a large country with widely dispersed health professionals. PMID- 16375797 TI - Web-based patient records and treatment guidelines in paediatric oncology. AB - We have established a Web-based system in Brazil offering health information in childhood cancer and services such as electronic patient records and treatment protocols. The system was based on open-source software. The database has records for about 3,200 patients from 30 Brazilian hospitals. An evaluation by doctors from the six hospitals comprising the Neuroblastoma Cooperative Group was positive, mainly because the system allows easy online access, but also because the electronic register performs data validation when information is inserted. Our experience shows that it is possible to use a Web-based system to provide paediatric cancer services at a distance. PMID- 16375798 TI - Information governance standards for managing e-health information. AB - Integrity of patient information, from both a quality and a security perspective, is critical to patient care. In the UK, the information governance initiative of the National Health Service (NHS) provides a framework to monitor and control the management of confidential patient data. Information governance standards grew out of the Data Accreditation Programme, first proposed in the 1998 NHS document Information for Health. The Data Accreditation Programme was based on a three stage assessment of data quality in acute hospitals. Stage one required internal review of policy and procedures for data input into computerized patient administration systems. Stage two involved an external audit to verify compliance with the standards. Stage three mandated audits of data outputs, focusing on clinical coding quality. Before stage three of the programme was fully implemented, the standards were incorporated into the information governance initiative, in which standards were expanded to include primary care and other health-care settings. These standards address many information management issues, including security and data quality, which are key concerns in telemedicine and e health applications. Compliance is essential for the successful implementation of the NHS Care Records Service, which will allow sharing of electronically stored patient information across the UK. PMID- 16375799 TI - Instant wireless transmission of radiological images using a personal digital assistant phone for emergency teleconsultation. AB - The instant transmission of radiological images may be important for making rapid clinical decisions about emergency patients. We have examined an instant image transfer system based on a personal digital assistant (PDA) phone with a built-in camera. Images displayed on a picture archiving and communication systems (PACS) monitor can be captured by the camera in the PDA phone directly. Images can then be transmitted from an emergency centre to a remote physician via a wireless high bandwidth network (CDMA 1 x EVDO). We reviewed the radiological lesions in 10 normal and 10 abnormal cases produced by modalities such as computerized tomography (CT), magnetic resonance (MR) and digital angiography. The images were of 24-bit depth and 1,144 x 880, 1,120 x 840, 1,024 x 768, 800 x 600, 640 x 480 and 320 x 240 pixels. Three neurosurgeons found that for satisfactory remote consultation a minimum size of 640 x 480 pixels was required for CT and MR images and 1,024 x 768 pixels for angiography images. Although higher resolution produced higher clinical satisfaction, it also required more transmission time. At the limited bandwidth employed, higher resolutions could not be justified. PMID- 16375800 TI - A tele-ultrasound needs analysis in Queensland. AB - A tele-ultrasound survey was distributed to clinicians and sonographers in regional and rural sites throughout Queensland. In all, 28 centres were invited to complete a profile questionnaire, and 27 centres were invited to complete a case-log. Twenty-five of the 28 questionnaires were returned (89% response rate). Twenty-two of the 27 case-logs were returned (81% response rate). Obstetric ultrasound was the most frequently performed ultrasound examination, accounting for 846 (35%) of the 2,410 recorded during the two-week survey period. The respondents considered that 182 cases would have benefited from further consultation: 79% for diagnostic advice, 13% for patient management advice and 8% for advice on examination technique. Six hospitals appeared to have the greatest need for a tele-ultrasound service. There was a strong preference for the store and-forward transmission of static images: 70% preferred this modality, in comparison with 19% who preferred realtime transmission and 11% who preferred store-and-forward transmission of video clips. PMID- 16375801 TI - Transmission of online digital documents for the health assessment component of e visa applications. AB - Up to 350,000 health assessments are conducted annually for persons seeking a visa to enter Australia. Health screening is conducted by accredited panel doctors and radiologists in the visa applicant's home country. Their findings are forwarded to the appropriate government department in Australia. The move to an e visa system, with certain visas being available via the Internet, has seen the introduction of a compatible electronic health assessment system. An online system to support health assessment processing was implemented in two stages. The first, eHealth1, has been in operation since July 2002 and the second, eHealth2, commenced in November 2003. In a pilot trial in Singapore during 2003 and 2004, 5,000 student visa applications (87%) were lodged through the eHealth2 system. The main advantage of the eHealth2 process is the rapid transmission of a digital photograph and X-ray image. This avoids the delay of sending documents and films by post. The success of the pilot has led to the expansion of eHealth2 to other electronic visa sectors, such as working holiday makers, long-stay business people and visitors. The eHealth2 system is fully operational in Singapore, Hong Kong, Japan, South Korea and Taiwan. PMID- 16375802 TI - Electronic patient record systems and the general practitioner: an evaluation study. AB - We studied how well electronic patient record (EPR) systems meet the needs of general practitioners (GPs) and other health-care professionals for specific information. GPs in eight health centres in the South-Ostrobothnia region of Finland were invited to participate. They used three types of EPR system. They were asked to access EPRs to obtain 20 types of information for patients receiving anticoagulant treatment. In total 2,389 patient cases were studied. All of the information requested was available for 73% of the cases (range 55-93%). There was a significant difference between the type of EPR system and the percentage of patients for whom information was available through the EPR. However, further analysis showed that differences in performance between EPR systems probably reflected differences in the way EPRs were used by different organizations. Great care should be taken in attempts to rank EPR systems based on their performance. PMID- 16375803 TI - Twenty years' experience of a telehealth service in the UK. AB - Since 1996 a small call centre has been operated to provide instant self-help, non-medical advice to members. Non-clinical advisers work under a medical director. The scheme grew to 1,200 member-families from a regional catchment area, peaking at 2,500 across the country. About 30-50 telephone calls are received each day and up to 1,000 Web visits per day; about 20-50 postal items are sent out daily to members. In a telephone poll during 2001, half of our current members said that they used the National Health Service about 70% less than before they joined, but a third said that they used complementary and alternative medicines 80% more. Half had gained in confidence by about 50% in managing their own health. Members most often join for alternative medical help with a specific problem, but stay if they begin to appreciate the help we can give them in regaining and keeping health in the round. Our retention of new members for a second year is only 10-20%. The 'barefoot' profession of telehealth adviser appears to meet a community need. PMID- 16375804 TI - Auto TeleCare -- understanding the failures and successes of small business in telehealth. AB - Auto TeleCare provided an automatic daily telephone service for people living alone. The business used an interactive voice response (IVR) system to call clients at a set time each day. The clients were required to press a button on their telephone to listen to a message (e.g. joke of the day), thereby indicating that they were alright. If the client did not respond, staff would call the given list of contacts to check on the client's welfare. The service was first offered in December 2003 and there was a lot of interest from clients and health-care groups. Although the technology was sophisticated, it was very simple for the clients to use. However, it was the marketing and advertising costs of the business that in the end proved to be too costly. The number of clients required for commercial viability was calculated to be 3,000, and after nearly 15 months of business it was decided to close the business. PMID- 16375805 TI - Case management and adherence to an online disease management system. AB - Non-adherence to treatment presents a significant obstacle to achieving favourable health outcomes. We have studied consumers' adherence to an online disease management system for depression, called Recovery Road. Recovery Road was implemented on a pilot basis for mental health care in Western Australia. Recovery Road was available for use by consumers and clinicians to augment usual treatment. One hundred and thirty consumers who had been diagnosed with major depression were enrolled. Consumers who used Recovery Road (n = 98) were provided with education, progress monitoring, e-consultation, e-diary and online evidenced based therapy. Consumers received either standard, automated adherence reminders by email (n = 69), or case management, which included personalized email and telephone follow-up in response to non-adherence (n = 29). After the first eight sessions, the adherence was 84% in the case management group and 55% in the automatic reminders group. The results suggest that case management increases adherence to online disease management systems. PMID- 16375806 TI - Telemedicine for rural and remote child and youth mental health services. AB - The E-Child and Youth Mental Health Service was designed to provide children and adolescents in Queensland with access to specialist mental health consultations using telemedicine. A project officer provided a single point of contact for referral management and clinic coordination, thereby reducing barriers of access to the service. Over a six-month period from November 2004, 42 point-to-point videoconferences were conducted to nine sites in Queensland. Three multipoint conferences were also conducted. Eleven videoconferences (24%) were arranged for administrative purposes, and 34 (76%) were conducted for the delivery of clinical services (30 patients). The referral and consultation activity suggests an improvement in the capacity of rural and remote mental health service providers to deliver specialist services for children and adolescents. PMID- 16375807 TI - An e-health needs assessment of medical residents in Cameroon. AB - Medical residents from Yaounde I University in Cameroon are required to spend periods of time in rural or remote locations to complete their training. To determine if e-health might lessen their isolation and enhance patient care, a needs assessment of the residents was performed using a brief questionnaire (five items) about the situation in which residents found themselves outside their medical school environment. We gave the questionnaires to 45 residents. Seventeen questionnaires had been returned at the time of the site visit, a response rate of 38%. Most residents indicated that the ability to contact a mentor would have either made them feel more confident (16, or 94%) or altered their handling of recent cases (15, or 88%). All residents had access to a mobile phone, and many (11, or 65%) had used it to contact a medical colleague for guidance. A low-cost and technologically simple telemedicine solution that maximized use of mobile phone capability, provided access to medical and health-care information, and permitted exchange of images would be an appropriate response to the identified needs. PMID- 16375808 TI - Wireless telemedicine for the delivery of specialist paediatric services to the bedside. AB - A mobile interactive online health system was used to conduct virtual ward rounds at a regional hospital which had no specialist paediatrician. The system was wireless, which allowed telepaediatric services to be delivered direct to the bedside. Between December 2004 and May 2005, 43 virtual ward rounds were coordinated between specialists based in Brisbane and local staff at the Gladstone Hospital. Eighty-six consultations were provided for 64 patients. The most common conditions included asthma (27%), chest infections (12%), gastroenteritis (10%) and urinary tract infections (10%). In the majority of cases, there were partial (67%) or complete changes (11%) in the clinical management of patients. Specialist services were offered by a team of 13 clinicians at the Royal Children's Hospital: 10 general paediatricians, two physiotherapists and one registered nurse. Feedback from all consultants involved in the service and local staff in Gladstone was extremely positive. In 43 videoconference calls there were three technical problems, probably due to an intermittent mains power supply at the regional hospital. There appears to be potential for other rural and regional hospitals to adopt this model of service delivery. PMID- 16375810 TI - A survey of e-mentoring among New Zealand midwives. AB - Mentoring is a strategy that may assist the midwifery profession to support new graduates and midwives working in rural and remote areas. We conducted a survey of 1,577 New Zealand midwives about their opinions and experiences of mentoring. The questionnaire comprised 33 questions, nine of which were open questions. There was a 44% response rate. While the telephone was commonly utilized by mentors (37%) and mentored midwives (37%), the Internet and email played only a small part. Participants acknowledged the potential of these avenues for communication, but midwives felt that mentoring could be provided only by immediate, face-to-face contact. Nevertheless, e-mentoring could be a viable option and requires further investigation. About one-third of midwives identified geographical isolation as a barrier to being a mentor (38%) and being mentored (36%). The use of e-mentoring could remove the barrier of location and allow the midwife to chose a mentor who meets her needs, rather than because she is the only mentor available. PMID- 16375809 TI - Realtime telemedicine for paediatric otolaryngology pre-admission screening. AB - We conducted a feasibility study to examine whether a paediatric patient at a regional hospital could be assessed by an ear, nose and throat (ENT) specialist via videoconference, therefore saving at least one journey to the tertiary hospital for a pre-admission appointment. A video-otoscope was used with standard videoconference equipment, and realtime images were transmitted at a bandwidth of 384 kbit/s. In all, 13 telepaediatric ENT clinics were conducted between November 2003 and April 2005, and 98 consultations were facilitated for 64 patients. The main reasons for referral were recurrent tonsillitis (25%) and obstructive sleep apnoea (23%). Of the 64 patients examined by telemedicine, 42 (66%) were recommended for surgery and placed on the surgical waiting list. About 12 patients (19%) required travel to the tertiary centre for further investigations and tests not available locally, while four patients (6%) were reviewed via videoconference during a scheduled clinic. Six patients (9%) required no further follow-up after their initial telepaediatric consultation. Videoconferencing is an effective method of assessing ENT conditions of paediatric patients and for pre-screening potential surgical admissions to a tertiary hospital. Careful consideration of a number of economic and logistical factors needs to be made before large investments are made to expand the service. PMID- 16375811 TI - User acceptance of the Web-based distribution of radiology services in regional and remote centres of Western Australia. AB - A large-scale, Web browser-based radiological information system integrated with a picture archiving and communication system (RIS/PACS) has been installed in regional and remote radiology centres of Western Australia. This provides patients in rural areas with local access to advanced medical imaging. User attitudes to the changes were surveyed in a six-week period starting in March 2005. A total of 53 people completed the survey (62% response rate). Respondents were eight radiologists, 21 medical imaging technologists, 10 administration officers and 14 reception staff. All groups gave average ratings of the system's quality. All staff rated the information quality of the system as good. On-site information technology support was judged to be reliable, professional and empathic towards user concerns. There was a broad range of opinion between groups, but on average they were satisfied. The lack of a modern network, comparable to what is available in the metropolitan areas, has impeded the full potential of RIS/PACS in the smaller rural centres. PMID- 16375812 TI - Supporting community carers via videoconferencing. AB - We provided support, practical advice and networking opportunities for carers in rural and remote areas of Western Australia through the use of videoconferencing. The first videoconference for carers occurred in June 2004. In the next 12 months there were 11 videoconferences with 11 rural towns. Some sessions were point-to point and some were multipoint. Over 90 carers participated in the sessions. A facilitator with expertise on stress in carers delivered the sessions from Perth. The acceptance of videoconferencing as a support medium by the carers attending was positive and there were requests for the sessions to be continued. The skills of the facilitator to connect in a very personal way with each person were important to the success of the programme. PMID- 16375813 TI - Successes and failures: what are we measuring? AB - In telemedicine and telehealth, the perception of success is complex, varies according to time and context, and depends on the perspective of the observer. Several reviews of the evaluation literature have been undertaken in recent years. These reviews identify common methodological shortcomings. Telehealth services continue to be funded as short-term projects. While it is essential to address methodology issues, it is important to understand that studies of pilot projects provide only interim findings about the feasibility of such applications, not how well they operate as mature applications. This represents something of a conundrum: evaluation is expected to establish the long-term value of telehealth using criteria which are specific to short-term projects. A useful approach would be to develop frameworks enabling all similar studies (e.g. diabetic home care) to be examined in order to extract commonalities and differences. This would enable us to draw conclusions about where telehealth is effective, as well as what variables demonstrate 'success'. PMID- 16375814 TI - Referral patterns in a global store-and-forward telemedicine system. AB - We examined the nature of the referral patterns in the email telemedicine network operated by the Swinfen Charitable Trust with a view to informing long-term resource planning. Over the first six years of operation, 62 hospitals from 19 countries registered with the Trust in order to be able to refer cases for specialist advice; 55 of these hospitals (89%) actually referred cases during this period. During the first six years of operation, nearly 1,000 referrals were submitted and answered, from a wide range of specialty areas. Between July 2002 and March 2005 the referral rate rose from 127 to 318 cases per year. The median length of time required to provide a specialist's response was 2.3 days during the first 12 months and 1.8 days during the last 12 months. Five hospitals submitted cases for more than four years (together sending a total of 493 cases). Their activity data showed a trend to declining referral rates over the four-year period, which may represent successful knowledge transfer. There is some evidence that over the last three years the growth in demand has been exponential, while the growth in resources available (i.e. specialists) has been linear, a situation which cannot continue for very long before demand outstrips supply. PMID- 16375815 TI - Work practice changes caused by the introduction of a picture archiving and communication system. AB - We studied the acceptance of work practice changes six months after the introduction of a picture archiving and communication system (PACS) in New South Wales. A one-page questionnaire was developed to assess doctors' and nurses' acceptance of work practice changes. Over 100 subjects were surveyed and 76 responded. In all, 92% of participants worked in the intensive-care unit (ICU) or the emergency department (ED). The results showed that the PACS had received a high level of acceptance. The respondents would not like to return to a film based practice. They were happy with the accessibility of images, especially when patients returned from an X-ray examination. At the time of survey, the doctors still had difficulty in remembering their user name and password. The users rarely or only sometimes used their own user names and passwords to log on the system. They were sometimes annoyed by the automatic time-out function. Nurses were happy that the PACS relieved their burden of searching for X-ray images for doctors. However, they distanced themselves from accessing the PACS, which they regarded as a doctors' tool. PMID- 16375817 TI - Individual and combined effects of low oral doses of deoxynivalenol and nivalenol in mice. AB - Deoxynivalenol (DON) and nivalenol (NIV) are toxic Fusarium secondary trichothecene metabolites that often co-occur regularly in cereal grains. These compounds were compared for their toxicity towards C57BL/6 mice on several parameters including alteration in plasma biochemistry, immune system reactivity and hepatic drug metabolism capacity. Mice received individual or combined oral doses of each toxin: 0.071 or 0.355 mg/kg of body weight, administrated three days a week for 4 weeks. Food consumption was altered by the single administration of 0.355 mg/kg of NIV, although no noticeable change of body and organ weights or liver protein contents was detected. NIV administration did cause also significant changes in total CO2 and uric acid concentrations in plasma. Individual toxin exposures led to increases in plasma IgA without no detectable change in the ex vivo production of cytokine by splenocytes. The liver ethoxyresorufin O-deealkylase, pentoxyresorufin O-depenthylase and glutathione S transferase activities were increased in concert with cytochrome P4501a and P4502b subfamily expression. Administration of combinations of DON and NIV resulted in responses similar to that observed using individual doses of each toxin. However, depending on the ratio of toxin doses and biochemical parameters, some responses could be also additive (plasma IgA and hepatic DCNB conjugation) or synergistic (plasma uric acid). PMID- 16375818 TI - Selective mineral elements concentration of the intestinal mucosa role of the lysosomes of duodenal enterocytes in the handling of mineral elements after intragastric administration. AB - Intragastric administration to rats of four soluble lanthanides cerium, lanthanum, europium, thulium and of three soluble elements of group IIIA aluminium, indium and gallium has been shown in previous studies. In this work two new rare earths gadolinium and terbium were studied using the same protocols and the same methods (transmission electron microscopy and ion microanalysis). among the previously studied elements, some of them were administered simultaneously on the one hand aluminium and indium, and on the other hand, lanthanum and cerium. These metals were looked for in intestinal mucosa, liver and kidney. The results showed: a) gadolinium and terbium were selectively concentrated in lysosomes of duodenal enterocytes, precipitated as non-soluble phosphate salts and eliminated with the cell's turn-over in less than 48 hr; b) Administered simultaneously, they precipitated in the same lysosome. c/ none of them was observed in the liver or kidney even with high dose. This study brings up to nine the number of elements forming a non-soluble phosphate salts, explaining their precipitation in lysosomes. None of them have a physiological role, two are toxic (aluminium and indium). This rapid intralysosomal concentration is an efficient mechanism which limits the diffusion of the foreign substances through the digestive barrier, then permits their elimination along with the cytoptose phenomenon in the intestinal lumen. PMID- 16375819 TI - Immunohistochemistry of adhesion molecules, metalloproteinases and NO-synthases in extravillous trophoblast of tubal pregnancy. AB - Trophoblast invasion in uterine pregnancy is fine-tuned for the remodelling of the uterine wall and its vascularization. Tubal pregnancy, which occurs in a limited number of patients, involves a dramatic trophoblast invasion in a context of a poor decidualization. By studying the histology of the extravillous trophoblast (EVC) in the anchoring villi, the Ki67 labelling, the location of several adhesion markers (cytokeratin-7, alpha1, alpha6, alphaV, beta1, beta4 integrin subunits and E-cadherin, V/E-cadherin), metalloproteinases (MMP-2, 9 and11), NOS2 and 3, we aimed to detect the specificity of tubal compared to intrauterine pregnancies. No difference could be observed between meso or anti salpingial trophoblast proliferation or invasion using Ki67. Cytokeratin-7 allowed detection of spindle-shape EVCs and we identified some decidualized stromal cells. Integrins alpha1, beta1 and alphaV, and V/E-cadherin were expressed mainly in the distal EVC correspondingly to intrauterine pregnancy, with a poor expression of alpha1. Integrins alpha6 and beta4, E-cadherin were detected in the distal EVC in contrast to uterine pregnancy. MMP-2, 9, 11 were also shown in distal EVC. NOS2 and 3 labelled the perivascular EVC and NOS3 the endothelial cells of the tubal vessels. These changed distributions of adhesion molecules and MMP together with that of the basic and inducible NOS expressions could be related to mechanical effects in superficial implantation or to a failure of decidualization in tubal pregnancies. PMID- 16375820 TI - The endothelin system and renin in human fetal membranes. AB - Term human fetal membranes express prorenin, a key enzyme within the renin angiotensin system. High levels of another vasoactive peptide, endothelin-1 (ET 1), are found in human amniotic fluid. To address the question of the relationship between these two vasoactive systems, we analyzed the expression of the components of the ET-1 system in fetal membranes in which cell types had been identified using different markers. Immunohistochemistry was performed with antibodies raised against the human proteins of the ET system. Term fetal membranes displayed ubiquitous labeling of endothelin-converting enzyme-1 (ECE-1) and ET-1. ETA receptors were detected in the chorionic connective tissue and the attached decidua; ETB receptors were localized to chorionic trophoblast cells and decidua. The localization of the ET-1 receptor subtype was confirmed by in-situ receptor binding. Renin immunoreactivity was detected in the chorionic connective tissue and the decidua. These findings suggest that ET-1 is produced ubiquitously in human fetal membranes, and its targets may be, trophoblast cells following ETB receptor activation, vascular structures and fibroblasts in the connective tissue and decidua via ETA and ETB receptors. It appears possible that renin and ET may contribute to the pathophysiological changes associated with premature labor and preeclampsia. PMID- 16375821 TI - Antioxidant status in alcohol-related diabetes mellitus in Beninese subjects. AB - In the present study, we investigated the antioxidant status in diabetes mellitus, related or not to alcohol consumption. A total of 38 type 1, 48 type 2 and 42 alcohol-related diabetic patients were selected. Total antioxidant status was assessed through the oxygen radical absorbance capacity of the plasma and the determination of enzymatic and non-enzymatic antioxidant molecules. Serum triglycerides, total cholesterol and HDL-cholesterol concentrations were determined and the lipid peroxydation was evaluated by measuring thiobarbituric acid reactive substances (TBARS) assay. Plasma total antioxidant capacity was more decreased in alcohol-related diabetes than that in type 1 and type 2 diabetes, regardless of the complications (retinopathy and renal failure). Plasma vitamin E concentrations were significantly decreased whereas those of vitamin C increased in all of the diabetic patients compared to the controls, irrespective to the complications. In addition, superoxide dismutase and glutathione peroxidase activities were reduced in all the patients (type 1, type 2 and alcohol-related), irrespective to the complications. Glutathione reductase activity was diminished in type 1 and alcohol-related, but not in type 2, diabetic patients. Glutathione (GSH) concentrations significantly decreased in all diabetic patients with a significant decrease in alcohol-related diabetic patients. Excessive alcohol consumption appears as an oxidative aggravating factor in diabetes mellitus. Besides, alcohol-related diabetes highly resembles to type 1 diabetes as far as the antioxidant parameters are concerned. PMID- 16375822 TI - 120 years of teaching microbiology at the University of Zagreb. PMID- 16375823 TI - Toxic effects of Ustilago maydis and fumonisin B1 in rats. AB - The toxicity of Ustilago maydis and the possible synergism with fumonisin B1 (FB1) were studied in Fischer rats by evaluating pathological changes and biochemical parameters in blood serum (LDH, ALT, GGT, ChE) and tissue homogenate of brain and liver (AChE, ChE, GGT, ALP). One experimental group (US) consumed diet with 70% of U. maydis galls and the other group (US+FB1) was fed pellets containing 70% of U. maydis galls and 1 mg of FB1 per kg of diet for 17 days. Control group (C) consumed standard pellets. During the trial, experimental animals were more excited, showing hyperactivity. Body mass gains slightly increased in both groups compared to the control. Gross pathological changes in liver, lungs, uterus and ovaries were more pronounced in the US+FB1 than in the US group. Specific catalytic activities of AChE decreased by 61% and by 63% in the liver and brain homogenate of the US group (p<0.05) compared to the control, indicating neurotoxic activity of U. maydis. Also, specific catalytic concentration of AChE and ALP was significantly decreased in the liver of the US+FB(1) group (p<0.05). Activity of LDH in the blood serum was increased up to 166% and 165% in the US+FB1 group (p<0.05) compared to the control and US group values, respectively, which indicates that FB1 was responsible for the disruption of cell membrane integrity. These findings suggest that Ustilago maydis and FB1 showed neurotoxicity in Fischer rats, which could be related to the alkaloids of U. maydis and disruption of sphingolipid metabolism by FB1 activity. PMID- 16375824 TI - Mould contamination and co-occurrence of mycotoxins in maize grain in Croatia. AB - Maize grain samples (n=15) collected during the autumn of 2002 were analyzed for the presence of moulds and mycotoxins fumonisin B1 (FB1), fumonisin B2 (FB2), zearalenone (ZEA), and ochratoxin A (OTA). Mycological analysis showed that all samples were contaminated with Fusarium spp. and Penicillium spp., while Aspergillus spp. were found in 5 samples. F. proliferatum and F. verticilloides, the producers of fumonisins, were found in 14 and 8 samples, respectively, while F. graminearum, the producer of ZEA, was present in all samples. The most frequent mycotoxins were FB1 (15/15) and ZEA (12/15), followed by OTA (7/15), while FB2 was found in only two samples. Seven samples were contaminated with two mycotoxins, seven with three, and one sample with only one mycotoxin. The concentrations (mean+/-SD) of FB1, ZEA, and OTA in positive samples were 459.5+/ 314.6, 1.70+/-0.80, and 1.40+/-0.55 microg kg-1, respectively, and the concentrations of FB2 in two samples were 68.4 and 3084.0 microg kg-1. In general, such low mycotoxin concentrations are not a significant source of exposure to humans, but they may contribute to exposure from other commodities. A few samples with extreme values indicate that strict control is needed. PMID- 16375825 TI - Verruculogen production in airborne and clinical isolates of Aspergillus fumigatus Fres. AB - Among airborne aspergilli sampled in outdoor air of the Zagreb area (2002/2003), Aspergillus niger (v. Teigh.) and A. fumigatus (Fres.) were the most abundant species (20-30%), with low mean annual concentrations (0.21-1.04 CFU m-3). Higher concentrations of A. fumigatus were observed in autumn and winter (0.5-1.05 CFU m 3) than in spring and summer (0-0.4 CFU m-3). On the other hand, A. fumigatus was found to be the most frequent isolate from upper and/or lower respiratory tracts of imunocompromised patients in many studies. This species produces several mycotoxins, including the tremorgenic mycotoxin verruculogen that can be found in spores and during myceliar growth. Verruculogen production ability was tested on 30 airborne and 33 clinical isolates of A. fumigatus. In both groups, high percentage of verruculogen-producing strains was noticed (84% of airborne and 91% of clinical isolates). Verruculogen production was not significantly different in the groups of airborne isolates (0.34+/-0.16 mg mL-1), and clinical isolates (0.26+/-0.19 mg mL-1). PMID- 16375826 TI - Mycotoxigenicity of clinical and environmental Aspergillus fumigatus and A. flavus isolates. AB - Clinical isolates of fifty strains of A. fumigatus and 30 strains of A. flavus from immmunocompromised patients from the hematological unit were analyzed for mycotoxin production and compared with the same number of environmental isolates (from soil, compost, and air). Only 9 (18%) strains of A. fumigatus produced gliotoxin in a mean concentration 2.22 mg mL-1 (range 0.5-5 mg mL-1). Aflatoxin B1 was detected in 7 (23%) isolates (range from 0.02 to 1.2 mg L-1) and aflatoxin G1 in one (3%) of clinical A. flavus isolates (0.12 mg L-1). In the group of environmental isolates, 11 (37%) were positive for aflatoxin B1 production (range from 0.02 to 1.2 mg L-1) and one for aflatoxin G1 (0.02 mg L-1). Bioautoantibiogram ("bioassay in situ") on TLC plates against Bacillus subtilis NCTC 8236 showed that only gliotoxin-producing strains have bactericidal activity of Rf values corresponding to gliotoxin. The secondary-metabolite profiles of clinical and environmental A. fumigatus and A. flavus isolates were homogeneous, except for gliotoxin production, which was detected only in the group of clinical isolates of A. fumigatus (18%). PMID- 16375827 TI - Antifungal activity of fluid extract and essential oil from anise fruits (Pimpinella anisum L., Apiaceae). AB - Antifungal activities of fluid extract and essential oil obtained from anise fruits Pimpinella anisum L. (Apiaceae) were tested in vitro on clinical isolates of seven species of yeasts and four species of dermatophytes. Diffusion method with cylinders and the broth dilution method were used for antifungal activity testing. Anise fluid extract showed antimycotic activity against Candida albicans, C. parapsilosis, C. tropicalis, C. pseudotropicalis and C. krusei with MIC values between 17 and 20% (v/v). No activity was noticed against C. glabrata, and anis fruits extracts showed growth promotion activity on Geotrichum spp. Anise fruits extract inhibited the growth of dermatophyte species (Trichophyton rubrum, T. mentagrophytes, Microsporum canis and M. gypseum) with MIC values between 1.5 and 9.0% (V/V). Anise essential oil showed strong antifungal activity against yeasts with MIC lower than 1.56% (V/V) and dermatophytes with MIC lower than 0.78% (V/V). Significant differences in antifungal activities were found between anise fluid extract and anise essential oil (p<0.01). Anise essential oil exhibited stronger antifungal activities against yeasts and dermatophytes with MIC values between 0.10 and 1.56% (V/V), respectively. PMID- 16375828 TI - Antimicrobial activity of N-phthaloylamino acid hydroxamates. AB - Antibacterial and antifungal activity of N-phthaloylamino acid hydroxamates [C6H4(CO)2N-X-CONHOH, X=amino acid residues of glycine, beta-alanine or D phenylglycine], was examined against 44 strains of Gram-positive and Gram negative bacteria, and 10 species of yeasts. The level of antimicrobial activity was established using the in vitro agar assay and the standard broth dilution susceptibility test. N-phthaloyl-D-phenylglycine-hydroxa- mic acid , the substance with the highest lipophilicity (log P), showed the best antibacterial activity, especially against Gram-negative bacteria. Minimum inhibitory concentration of was 0.008 mg mL-1 in the activity against Yersinia enterocolitica O3, confirmed by a large inhibition zone (30 mm) by the diffusion test. Hydroxamates inhibit growth by chelation of the PDF enzyme metal in both Gram-positive and Gram-negative bacteria, and LpxC enzyme in Gram-negative enzyme. Phthalimides appear to contribute to inhibition by destabilizing m-RNA. Antifungal activity of substances is not very expressed. PMID- 16375829 TI - Antimicrobial activity of some hydroxamic acids. AB - Several hydroxamic acids, viz., N-benzyl-N'-hydroxysuccinamide (BHS), poly[alpha,beta-(N-hydroxy)-DL-aspartamide] (PHA), poly[alpha,beta-(N-hydroxy-N methyl-DL-aspartamide)] (PMHA) and poly[alpha,beta-(N-hydroxy)-DL- aspartamide]/poly[alpha,beta-(N-2-hydroxyethyl)-DL-aspartamide] (PHA-PHEA 1:1) were prepared and screened for their antimicrobial activity. Ten Gram-positive and 7 Gram-negative species of bacteria, 5 Candida species, 4 dermatophyte species and 3 mould species were used in tests. Compound showed no antimicrobial activity on any of the tested microorganisms. Other compounds showed a narrow spectrum of antibacterial activity, but no antifungal activity. PMID- 16375830 TI - Investigation of antimicrobial activity of Pelargonium radula (Cav.) L'Herit. AB - Antimicrobial activities of two ethanolic extracts, made from fresh and dried leaves of Pelargonium radula (Cav.) L'Herit, were tested against fourteen species of bacteria and fifteen species of fungi. The well-diffusion method indicated the strongest activity against Pseudomonas aeruginosa. The broth dilution method revealed that the most sensitive microorganisms were Bacillus pumilus, Bacillus subtilis, Escherichia coli and Serratia marcescens. Extract prepared from fresh leaves showed significantly higher antimicrobial activity than the extract prepared from dried leaves. PMID- 16375831 TI - Antimicrobial activity of juniper berry essential oil (Juniperus communis L., Cupressaceae). AB - Juniper essential oil (Juniperi aetheroleum) was obtained from the juniper berry, and the GC/MS analysis showed that the main compounds in the oil were alpha pinene (29.17%) and beta-pinene (17.84%), sabinene (13.55%), limonene (5.52%), and mircene (0.33%). Juniper essential oil was evaluated for the antimicrobial activity against sixteen bacterial species, seven yeast-like fungi, three yeast and four dermatophyte strains. Juniper essential oil showed similar bactericidal activities against Gram-positive and Gram-negative bacterial species, with MIC values between 8 and 70% (V/V), as well as a strong fungicidal activity against yeasts, yeast-like fungi and dermatophytes, with MIC values below 10% (V/V). The strongest fungicidal activity was recorded against Candida spp. (MIC from 0.78 to 2%, V/V) and dermatophytes (from 0.39 to 2%, V/V). PMID- 16375832 TI - Flavonoid analysis and antimicrobial activity of commercially available propolis products. AB - Propolis ethanolic solutions are the most used propolis products on the market for the treatment of minor ulcers in the mouth, angina, thrush or skin infections. Since it is still an unofficial drug in pharmacy, we analyzed the contents of flavonoids in ten commercially available ethanolic solutions of propolis from the Croatian market using two complementary colorimetric methods. Antimicrobial activities, determined with the diffusion method, against six bacterial species (Bacillus subtilis NCTC 8236, Staphylococcus aureus ATCC 25923, Streptococcus pyogenes ATCC 12204, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 10536, Pseudomonas aeruginosa ATCC 27853, and one yeast-like fungus Candida albicans ATCC 10231 were compared. Results of flavonoids analysis suggested that the contents of flavones and flavonols in the products were uniform and ranged from 0.14 to 0.41%, but the content of flavanones varied greatly from 0.43 to 18.78%. Total flavonoid content, as the sum of two colorimetric methods, in propolis products was between 0.78 and 18.92%, and most products had the flavonoids content below 9%. All products with the total flavonoids content above 1% showed antimicrobial activity against the four Gram positive bacterial species tested, and against P. aeruginosa and the yeast-like fungus C. albicans. Total flavonoids contents, expressed as the sum of two colorimetric methods, could be useful methods for estimating the flavonoid contents of propolis products. Our results indicate that the quality of commercially available propolis products requires verification. PMID- 16375833 TI - Antimicrobial activity of flavonoids from Pelargonium radula (Cav.) L'Herit. AB - Flavonoids from Pelargonium radula (Cav.) L'Herit were purified by column chromatography. Two fractions were obtained: F1 (main flavonoid isoquercitrin) and F2 (main flavonoid rutin). In vitro antimicrobial activity of F1 and F2 were tested against eleven species of bacteria and eleven species of fungi. Both fractions demonstrated strong inhibitory activity against Staphylococcus aureus, Proteus rettgeri, Candida tropicalis and Microsporum gypseum. Staphylococcus sp. (coagulase-negative) and Candida lusitaniae were strongly inhibited only by fraction F1 and Fusarium graminearum only by fraction F2. PMID- 16375834 TI - Influence of cyclodextrin complexation on piroxicam gel formulations. AB - The aim of this work was to evaluate the role of cyclodextrins in topical drug formulations. Solid piroxicam (PX) complexes with beta-cyclodextrin (beta-CD) and randomly methylated beta-cyclodextrin (RAMEB) were prepared by freeze-drying and characterized using differential scanning calorimetry (DSC), X-ray powder diffractometry (XRPD), Fourier transform infrared spectroscopy (FTIR) and near infrared spectroscopy (NIR). A physical mixture of PX and cyclodextrins was characterized by enhanced dissolution properties compared to the dissolution profile of the pure drug due to in situ complex formation. Formation of the PX cyclodextrin inclusion complex additionally improved the drug dissolution properties. Influence of CDs on drug permeation from the water dispersion and the prepared hydroxypropyl methylcellulose (HPMC) gels was investigated. Permeation of the drug involved three consecutive processes: dissolution of the solid phase, diffusion across the swollen polymer matrix and drug permeation through the membrane. Complexation increased PX diffusion by increasing the amount of diffusible species in the donor phase. Slower drug diffusion through the HPMC matrix was the rate limiting step in the overall diffusion process. Possible interaction between the hydrophilic polymer and cyclodextrin may result in physicochemical changes, especially in a change of rheological parameters. PMID- 16375835 TI - Synthesis and antimicrobial screening of some fused heterocyclic pyrroles. AB - Pyrrole derivatives 1a,b were used as precursors for the preparation of pyrrolo[2,3-d]pyrimidine derivatives 2a,b-7a,b. Also, the formation and structure of different pyrrolotriazolopyrimidine derivatives 8a,b-11a,b were discussed. Some of the prepared products showed potent antimicrobial activity. PMID- 16375836 TI - Formulation and evaluation of diazepam hydrogel for rectal administration. AB - Diazepam (DZP) has become a commonly used drug for treatment of acute repetitive epileptic seizures and febrile convulsions in children. Considering the advantages of rectal administration of DZP, the objective of our study was to formulate and evaluate rectal hydrogels containing DZP as a drug substance in combination with suitable co-solvents and preservatives. Prepared HPMC (hydroxypropyl methylcellulose) hydrogels containing different concentrations of DZP (2, 4 and 6 mg mL(-1)) manifested good quality in respect to physico-chemical parameters (pH value, drug content, ingredients content and viscosity), antimicrobial efficiency and microbiological quality. Under the proposed HPLC conditions, satisfactory separation of DZP and the preservatives used was achieved. In vitro release studies have shown that the total amount of DZP was released in a period of 3 h. Prepared formulations were stable for four months at 26 degrees C (ambient temperature characteristic of the 2nd climate zone). PMID- 16375837 TI - Assessment of Albizia zygia gum as a binding agent in tablet formulations. AB - Albizia gum has been evaluated as a binding agent in tablet formulations in comparison with gelatin BP. Compressional properties were analyzed using density measurements and the compression equations of Heckel and Kawakita as assessment parameters, while the mechanical properties of the tablets were assessed using the crushing strength and friability of the tablets. Drug release properties of the tablets were assessed using disintegration time and dissolution time as assessment parameters. Formulations containing Albizia gum as a binding agent show a faster onset and higher amount of plastic deformation under compression pressure than those containing gelatin. The crushing strength, disintegration and dissolution times of the tablets increased with increased binder concentration while their friability decreased. Albizia gum produced tablets with better mechanical properties and longer disintegration and dissolution times than those containing gelatin BP. This suggests that Albizia gum could be useful as a binding agent especially when high mechanical strength and slower release rates are desired. PMID- 16375838 TI - Floating microspheres of cimetidine: formulation, characterization and in vitro evaluation. AB - The present study involves preparation and evaluation of floating microspheres with cimetidine as model drug for prolongation of gastric residence time. The microspheres were prepared by the solvent evaporation method using polymers hydroxypropylmethyl cellulose and ethyl cellulose. The shape and surface morphology of prepared microspheres were characterized by optical and scanning electron microscopy, respectively. In vitro drug release studies were performed and drug release kinetics was evaluated using the linear regression method. Effects of the stirring rate during preparation, polymer concentration, solvent composition and dissolution medium on the size of microspheres and drug release were also observed. The prepared microspheres exhibited prolonged drug release (approximately 8 h) and remained buoyant for > 10 h. The mean particle size increased and the drug release rate decreased at higher polymer concentration. No significant effect of the stirring rate during preparation on drug release was observed. In vitro studies demonstrated diffusion-controlled drug release from the microspheres. PMID- 16375839 TI - Synthesis of some oxazolinones and imidazolinones and their antimicrobial screening. AB - A few imidazolinones [1-aminoethyl/phenyl-2-methyl/phe- nyl-4 acetylidene/benzylidene-imidazolin-5[4H]-ones] were newly synthesized from respetive acetylidene/benzylidene oxazolinones. Schiff's bases were synthesized by the reaction between imidazolinones and benzaldehyde. The antimicrobial screening of almost all compounds showed moderate to significant activities against B. subtilis ATCC 6633 and K. pneumoniae ATCC 25063. Compounds 10 [1 aminophenyl-2-phenyl-4-acetylidene-imidazolin-5[4H]-one] and 12 [1-aminophenyl-2 phenyl-4-benzylidene-imi- dazolin-5[4H]-one] showed even better activity than amphotericin B against C. albicans ATCC 29738. PMID- 16375840 TI - Free radical scavenging potential of Saussarea costus. AB - Saussurea costus (Falc.) Lipschitz from the family Asteraceae is an important medicinal drug, the roots of which are widely used in folk medicine. The antioxidant activity of the plant has been studied using its ability to scavenge DPPH, nitric oxide, superoxide radicals along with its ability to inhibit lipid peroxidation and GSH oxidation. The 1 mg mL(-1) extract had antioxidant activity with 85.2% reduction of DPPH and a 72.7% decrease in lipid peroxidation. It showed maximum inhibition of superoxide radical of 66.0%, and 58.4% inhibition of nitric oxide formation. The concentration of chlorogenic acid was 0.027% in the extract of S. costus. Thus, the therapeutic activity of the plant may be due to its antioxidant activity, probably as a result of the presence of chlorogenic acid. PMID- 16375841 TI - Ocular inserts for controlled delivery of pefloxacin mesylate: preparation and evaluation. AB - Pefloxacin mesylate is a flouroquinolone antibacterial drug effective in the treatment of bacterial conjunctivitis. The objective of the present work was to develop ocular inserts of pefloxacin mesylate and evaluate their potential for sustained ocular delivery. Reservoir-type ocular inserts were prepared by the film casting technique in teflon coated Petri dishes and characterized in vitro by drug release studies using a flow-through apparatus that simulated the eye conditions. Six formulations were developed, which differed in the ratio of polymers Eudragit RS 100 and Eudragit RL 100 used for the preparation of the rate controlling membrane. All formulations carried 0.72 mg pefloxacin mesylate, 2.69 mg polyvinyl pyrrolidone (PVP) K-30, plasticizers, propylene glycol (10% m/m) and dibutyl phthalate (15%, m/m). The optimized formulation was subjected to microbiological studies, in vivo studies, interaction studies, and stability studies to assess the effectiveness of the formulation. Cumulative drug released from the formulation ranged from 90-98% within 48 to 120 hours. On the basis of in vitro drug release studies, the formulation with Eudragit RS 100/Eudragit RL 100 (4:1) was found to be better than the other formulations and it was selected as an optimized formulation. On the basis of in vitro, microbiological, in vivo drug release, interaction and stability studies, it can be concluded that this ocular insert formulation provided the desired drug release in vitro for 5 days and remained stable and intact at ambient conditions. PMID- 16375842 TI - Chemical composition and antibacterial activity of the rhizome oil of Hedychium larsenii. AB - The composition of essential oil from the rhizomes of Hedychium larsenii M. Dan & Sathish was examined by GC-FID and GC-MS techniques. 99% of the oil consisted of monoterpenoids. Sesquiterpenoids were present only in negligible quantities. Linalool and 1,8-cineole were identified as the major components. The oil showed moderate antibacterial activity against Gram-positive and Gram-negative bacteria. PMID- 16375843 TI - Essential oil of the Persian sage, Salvia rhytidea Benth. AB - Chemical composition of volatile compounds from Salvia rhytidea Benth. was analyzed, for the first time, by gas chromatography/mass spectrometry. The volatiles were isolated from dried aerial parts of the plant by hydrodistillation. A total yield of 2.0 mg of essential oil per g of plant dry mass was obtained and sixty compounds were identified, representing 98.2% of total volatiles. The essential oil was characterized by a high content of hydrocarbon and oxygenated monoterpenes. The main constituents were p-cymene-8-ol (11.9%), spathulenol (7.3%), pulegone (6.4%), sabinene (5.8%), terpinen-4-ol (5.5%) and alpha-copaene (5.3%). PMID- 16375844 TI - Risky driving and lifestyles in adolescence. AB - Several studies have shown that risky driving is especially prevalent among young drivers and recent research has pointed out that driving in adolescence should be investigated in the more general context of adolescent development. The first aim of this contribution was to analyze involvement in risky driving in a normative sample of 645 Italian adolescents, boys and girls, aged 14-17, through a self report questionnaire. A second aim was to evaluate the association between risky driving and lifestyle, defined as involvement in other health risk behaviors and leisure activities. The main results showed that many adolescents drove cars and motorcycles without the required driving license and the most frequent offences were speeding and failure to maintain a safe braking distance. Gender and age differences were also investigated. Results concerning the association between risky driving and lifestyle showed that risky driving was not an isolated behavior. Boys who displayed risky driving practices were more likely to adopt a lifestyle characterized by high involvement in antisocial behaviors, tobacco smoking, comfort eating and time spent in non-organized activities with friends. Girls involved in risky driving were more likely to be involved in other risk taking behaviors, antisocial behaviors and drug use. PMID- 16375845 TI - [Are preoperative examinations useful in the management of patients with renal hyperparathyroidism?]. AB - AIM OF THE STUDY: To evaluate the efficiency of preoperative parathyroid ultrasonography and scintigraphy in the management of renal hyperparathyroidism. PATIENTS AND METHODS: The charts of the last consecutive 200 patients who underwent surgery for renal hyperparathyroidism from 1998 to 2003 were retrospectively reviewed to collect data concerning parathyroid gland function, results of preoperative ultrasonography and scintigraphy, as well as modalities and results of surgical exploration. RESULTS: Ultrasonography and scintigraphy sensibilities were 36.4% and 49.3%, respectively. Efficiency of both examinations was improved when they were combined (sensibility of 64.7%) and in those patients managed for recurrent hyperparathyroidism. Were more often detected by preoperative examinations glands with high weight and/or greatest diameter, orthotopic and inferior glands as well as glands exhibiting nodular hyperplasia content upon pathological examination. CONCLUSION: Parathyroid ultrasonography and scintigraphy are of poor interest in the management of renal hyperparathyroidism. In a preoperative setting, they should be performed only in patients with recurrent disease. PMID- 16375846 TI - Heme-protein covalent bonds in peroxidases and resistance to heme modification during halide oxidation. AB - Plant peroxidases, as typified by horseradish peroxidase (HRP), primarily catalyze the one-electron oxidation of phenols and other low oxidation potential substrates. In contrast, the mammalian homologues such as lactoperoxidase (LPO) and myeloperoxidase primarily oxidize halides and pseudohalides to the corresponding hypohalides (e.g., Br(-) to HOBr, Cl(-) to HOCl). A further feature that distinguishes the mammalian from the plant and fungal enzymes is the presence of two or more covalent bonds between the heme and the protein only in the mammalian enzymes. The functional roles of these covalent links in mammalian peroxidases remain uncertain. We have previously reported that HRP can oxidize chloride and bromide ions, but during oxidation of these ions undergoes autocatalytic modification of its heme vinyl groups that virtually inactivates the enzyme. We report here that autocatalytic heme modification during halide oxidation is not unique to HRP but is a general feature of the oxidation of halide ions by fungal and plant peroxidases, as illustrated by studies with Arthromyces ramosus and soybean peroxidases. In contrast, LPO, a prototypical mammalian peroxidase, is protected from heme modification and its heme remains intact during the oxidation of halide ions. These results support the hypothesis that the covalent heme-protein links in the mammalian peroxidases protect the heme from modification during the oxidation of halide ions. PMID- 16375847 TI - Biosynthetic potential of sesquiterpene synthases: alternative products of tobacco 5-epi-aristolochene synthase. AB - Nicotiana tabacum (tobacco) 5-epi-aristolochene synthase (TEAS) serves as an useful model for understanding the enzyme mechanisms of sesquiterpene biosynthesis. Despite extensive bio-chemical and structural characterization of TEAS, a more detailed analysis of the reaction product spectrum is lacking. This study reports the discovery and quantification of several alternative sesquiterpene products generated by recombinant TEAS in the single-vial GC-MS assay. The combined use of chiral and non-polar stationary phases for gas chromatography separations proved critical for resolving the numerous sesquiterpene products of TEAS for mass spectral analysis and identification. Co injection studies with available authentic standards from both synthetic and natural sources further corroborated the assignment of several compounds, resulting in an annotated reaction mechanism accounting for their biosynthesis. Moreover, a previously undocumented farnesyl trans-cis isomerization pathway was observed. PMID- 16375848 TI - Acute effect of fatty acids on metabolism and mitochondrial coupling in skeletal muscle. AB - Acute effects of free fatty acids (FFA) were investigated on: (1) glucose oxidation, and UCP-2 and -3 mRNA and protein levels in 1 h incubated rat soleus and extensor digitorium longus (EDL) muscles, (2) mitochondrial membrane potential in cultured skeletal muscle cells, (3) respiratory activity and transmembrane electrical potential in mitochondria isolated from rat skeletal muscle, and (4) oxygen consumption by anesthetized rats. Long-chain FFA increased both basal and insulin-stimulated glucose oxidation in incubated rat soleus and EDL muscles and reduced mitochondrial membrane potential in C2C12 myotubes and rat skeletal muscle cells. Caprylic, palmitic, oleic, and linoleic acid increased O(2) consumption and decreased electrical membrane potential in isolated mitochondria from rat skeletal muscles. FFA did not alter UCP-2 and -3 mRNA and protein levels in rat soleus and EDL muscles. Palmitic acid increased oxygen consumption by anesthetized rats. These results suggest that long-chain FFA acutely lead to mitochondrial uncoupling in skeletal muscle. PMID- 16375849 TI - A theoretical study on nitric oxide reductase activity in a ba(3)-type heme copper oxidase. AB - The mechanism of nitric oxide reduction in a ba(3)-type heme-copper oxidase has been investigated using density functional theory (B3LYP). Four possible mechanisms have been studied and free energy surfaces for the whole catalytic cycle including proton and electron transfers have been constructed by comparison to experimental data. The first nitric oxide coordinates to heme a(3) and is partly reduced having some nitroxyl anion character ((3)NO(-)), and it is thus activated toward the attack by the second N-O. In this reaction step a cyclic hyponitrous acid anhydride intermediate with the two oxygens coordinating to Cu(B) is formed. The cyclic hyponitrous acid anhydride is quite stable in a local minimum with high barriers for both the backward and forward reactions and should thus be observable experimentally. To break the N-O bond and form nitrous oxide, the hyponitrous acid anhydride must be protonated, the latter appearing to be an endergonic process. The endergonicity of the proton transfer makes the barrier of breaking the N-O bond directly after the protonation too high. It is suggested that an electron should enter the catalytic cycle at this stage in order to break the N-O bond and form N(2)O at a feasible rate. The cleavage of the N-O bond is the rate limiting step in the reaction mechanism and it has a barrier of 17.3 kcal/mol, close to the experimental value of 19.5 kcal/mol. The overall exergonicity is fitted to experimental data and is 45.6 kcal/mol. PMID- 16375850 TI - Dinitrophenol-induced mitochondrial uncoupling in vivo triggers respiratory adaptation in HepG2 cells. AB - Here, we show that 3 days of mitochondrial uncoupling, induced by low concentrations of dinitrophenol (10 and 50 microM) in cultured human HepG2 cells, triggers cellular metabolic adaptation towards oxidative metabolism. Chronic respiratory uncoupling of HepG2 cells induced an increase in cellular oxygen consumption, oxidative capacity and cytochrome c oxidase activity. This was associated with an upregulation of COXIV and ANT3 gene expression, two nuclear genes that encode mitochondrial proteins involved in oxidative phosphorylation. Glucose consumption, lactate and pyruvate production and growth rate were unaffected, indicating that metabolic adaptation of HepG2 cells undergoing chronic respiratory uncoupling allows continuous and efficient mitochondrial ATP production without the need to increase glycolytic activity. In contrast, 3 days of dinitrophenol treatment did not change the oxidative capacity of human 143B.TK(-) cells, but it increased glucose consumption, lactate and pyruvate production. Despite a large increase in glycolytic metabolism, the growth rate of 143B.TK(-) cells was significantly reduced by dinitrophenol-induced mitochondrial uncoupling. We propose that chronic respiratory uncoupling may constitute an internal bioenergetic signal, which would initiate a coordinated increase in nuclear respiratory gene expression, which ultimately drives mitochondrial metabolic adaptation within cells. PMID- 16375851 TI - Adsorption equilibria between liposome membrane formed of phosphatidylcholine and aqueous sodium chloride solution as a function of pH. AB - The effect has been studied of the adsorption of ions (H(+), Na(+), OH(-), Cl(-)) which are present in solution upon the electric charge of the liposome membrane formed of phosphatidylcholine (PC). The surface charge density of the membrane was determined as a function of pH and electrolyte concentration from electrophoretic mobility measurements. The measurements were carried out by the laser-Doppler microelectrophoresis method. A four-equilibria model has been proposed to describe the phenomena occurring on the membrane surface. The equilibria in which the adsorption of other ions on the liposome membrane surface was involved were assumed to exist beside the equilibria in which the H(+) and OH(-) ions were engaged. The idea was confirmed by mathematical calculations. Association constants of the liposome membrane surface with ions of solution (K(AH), K(ANa), K(BOH), K(BCl)) were determined. The proposed model has been proved to be correct by comparing the resulting theoretic charge variation curves of the lecithin membrane with the experimental data. PMID- 16375852 TI - The role of extinction and reinstatement in attentional bias to threat: a conditioning approach. AB - We investigated the effects of extinction and reinstatement on attentional bias to fear-conditioned signals in healthy individuals using an emotional modification of a spatial cueing paradigm. Spatial cues were emotionally modulated using differential conditioning. The CS+ was sometimes followed by an aversive electrocutaneous stimulus (UCS), whereas the CS- was never followed by the UCS. During a subsequent extinction phase no UCS was presented anymore. The reinstatement phase started with one or four unpredicted UCS-only trials for half of the participants (reinstatement group). For the other half there were no additional UCS presentations (control group). We found that attention was biased to threat signals during acquisition. This biased attention largely disappeared during extinction. During the reinstatement phase attentional bias to threat signals re-emerged in the reinstatement group, but not in the control group. PMID- 16375853 TI - Renewal of cue-elicited urge to smoke: implications for cue exposure treatment. AB - The effects of cue exposure therapy are limited, because renewal after extinction is an important source of relapse. In this study, 33 smokers were exposed to a cue predicting smoking availability and a cue predicting smoking unavailability in one context (acquisition context A). Following extinction in another context (extinction context B), a test for renewal took place in the original acquisition context A (i.e. ABA renewal). Urge to smoke was measured using a Visual Analogue Scale. Renewal of differential conditioned urge responding occurred when participants were tested in the acquisition context, while differential urge responding remained extinguished when tested in the extinction context. This experiment provides evidence that ABA renewal occurred in smokers. Clinical implications are discussed. PMID- 16375854 TI - Overexpression of HMGA2-LPP fusion transcripts promotes expression of the alpha 2 type XI collagen gene. AB - In a subset of human lipomas, a specific t(3;12) chromosome translocation gives rise to HMGA2-LPP fusion protein, containing the amino (N)-terminal DNA binding domains of HMGA2 fused to the carboxyl (C)-terminal LIM domains of LPP. In addition to its role in adipogenesis, several observations suggest that HMGA2-LPP is linked to chondrogenesis. Here, we analyzed whether HMGA2-LPP promotes chondrogenic differentiation, a marker of which is transactivation of the alpha 2 type XI collagen gene (Col11a2). Real-time PCR analysis showed that HMGA2-LPP and COL11A2 were co-expressed. Luciferase assay demonstrated that either of HMGA2 LPP, wild-type HMGA2 or the N-terminal HMGA2 transactivated the Col11a2 promoter in HeLa cells, while the C-terminal LPP did not. RT-PCR analysis revealed that HMGA2-LPP transcripts in lipomas with the fusion were 591-fold of full-length HMGA2 transcripts in lipomas without the fusion. These results indicate that in vivo overexpression of HMGA2-LPP promotes chondrogenesis by upregulating cartilage-specific collagen gene expression through the N-terminal DNA binding domains. PMID- 16375855 TI - Calcium-dependent movement of troponin I between troponin C and actin as revealed by spin-labeling EPR. AB - We measured EPR spectra from a spin label on the Cys133 residue of troponin I (TnI) to identify Ca(2+)-induced structural states, based on sensitivity of spin label mobility to flexibility and tertiary contact of a polypeptide. Spectrum from Tn complexes in the -Ca(2+) state showed that Cys133 was located at a flexible polypeptide segment (rotational correlation time tau=1.9ns) that was free from TnC. Spectra of both Tn complexes alone and those reconstituted into the thin filaments in the +Ca(2+) state showed that Cys133 existed on a stable segment (tau=4.8ns) held by TnC. Spectra of reconstituted thin filaments (-Ca(2+) state) revealed that slow mobility (tau=45ns) was due to tertiary contact of Cys133 with actin, because the same slow mobility was found for TnI-actin and TnI tropomyosin-actin filaments lacking TnC, T or tropomyosin. We propose that the Cys133 region dissociates from TnC and attaches to the actin surface on the thin filaments, causing muscle relaxation at low Ca(2+) concentrations. PMID- 16375856 TI - Dominant role of copper in the kinetic stability of Cu/Zn superoxide dismutase. AB - Mutations in Cu/Zn superoxide dismutase (SOD) are involved in some cases of familial amyotrophic lateral sclerosis, and it appears that misfolding and aggregation, perhaps mediated by abnormal binding or loss of copper (Cu) and/or zinc (Zn), may play a pathological role. It is known that the absence of both metals kinetically destabilizes wild type and mutant SOD leading to a 60-fold increase in their rate of unfolding. Here, the individual contributions of Cu and Zn to the kinetic stability of SOD were investigated, and the results show that Cu plays a greater role. Thus, the deficiency of Cu or Zn, especially the former, will compromise the kinetic stability of SOD, thereby increasing the probability that pathogenic mutants and even the WT protein may misfold and self-assemble into toxic species. PMID- 16375857 TI - Glucose activation of ChREBP in hepatocytes occurs via a two-step mechanism. AB - Carbohydrate response element binding protein (ChREBP) is a transcription factor that mediates glucose-responsive changes in gene expression in hepatocytes. In the current model for glucose regulation, inhibition of ChREBP in low glucose occurs in response to cAMP-dependent protein kinase (PKA)-mediated phosphorylation of residues S196, S626, and/or T666. Activation of ChREBP in conditions of increased glucose results simply from reversal of these inhibitory phosphorylations. To test this model, we analyzed mutant forms of ChREBP that lack one or more of the proposed PKA sites and found that these forms of ChREBP still require glucose for activation. Additionally, cAMP levels in cultured hepatocytes were negligible in low glucose conditions, indicating PKA should not be active. Finally, overall ChREBP phosphorylation did not change in response to altered glucose levels. We conclude that in addition to its repression by PKA, glucose activation of ChREBP involves a second mechanism that is independent of PKA phosphorylation. PMID- 16375858 TI - Involvement of rho kinase in the ouabain-induced contractions of the rat renal arteries. AB - Agonist and depolarization-induced vascular smooth muscle contractions include the activation of rho/rho kinase pathway. However, there are no reports addressing the question whether this pathway is involved in ouabain-induced vascular smooth muscle contractions. Therefore, in this study, the possible participation of the rho/rho kinase pathway in ouabain-induced contractions was evaluated in rat renal arteries. Effects of rho kinase inhibitors (fasudil and Y 27632) on ouabain-induced contractions, and phosphorylation of myosin binding subunits (MYPT/MBS85) of myosin phosphatase were determined using isolated tissue and Western blot experiments, respectively. Fasudil and Y-27632 inhibited ouabain induced contractions in a concentration-dependent manner. The phosphorylation of MYPT was not altered by ouabain. However, ouabain significantly increased MBS85 phosphorylation of myosin phosphatase. The phosphorylation of both subunits of myosin phosphatase was inhibited by Y-27632. These results indicate that activation of rho kinase and the subsequent phosphorylation of MBS85 are involved in ouabain-induced contraction of rat renal arteries. This mechanism may be important in essential hypertension with elevated endogenous ouabain levels. PMID- 16375859 TI - Ascorbic acid reduction of microtubule protein disulfides and its relevance to protein S-nitrosylation assays. AB - The biotin switch assay was developed to aid in the identification of S nitrosylated proteins in different cell types. However, our work with microtubule proteins including tubulin and its associated proteins tau and microtubule associated protein-2 shows that ascorbic acid is not a selective reductant of protein S-nitrosothiols as described in the biotin switch assay. Herein we show that ascorbic acid reduces protein disulfides in tubulin, tau, and microtubule associated protein-2 that are formed by peroxynitrite anion. Reduction of microtubule-associated protein disulfides by ascorbic acid following peroxynitrite treatment restores microtubule polymerization kinetics to control levels. We also show that ascorbic acid reduces the disulfide dithiobis(2 nitrobenzoic acid), a reagent commonly used to detect protein thiols. Not only do we describe a new reactivity of ascorbic acid with microtubule proteins but we expose an important limitation when using the biotin switch assay to detect protein S-nitrosylation. PMID- 16375860 TI - Smilaxin, a novel protein with immunostimulatory, antiproliferative, and HIV-1 reverse transcriptase inhibitory activities from fresh Smilax glabra rhizomes. AB - A protein, with a novel N-terminal amino acid sequence and a molecular mass of 30 kDa, was purified from fresh Smilax glabra rhizomes by adsorption on DEAE cellulose, CM-cellulose, Con A-Sepharose, and Mono S, and by fast protein liquid chromatography-gel filtration on Superdex 75. The protein, designated as smilaxin, stimulated uptake of [methyl-3H]thymidine by murine splenocytes, peritoneal macrophages, and bone marrow cells, and production of nitric oxide by peritoneal macrophages. It inhibited uptake of [methyl-3H]thymidine by MBL2 and PU5 tumor cells but not uptake by S180 and L1210 cells. Smilaxin augmented glucose uptake into rat adipose tissue. It attenuated the activity of HIV-1 reverse transcriptase with an IC50 of 5.6 microM. However, it did not display hemagglutinating, antifungal or translation-inhibitory activities, indicating that it is not a lectin, an antifungal protein, or a ribosome-inactivating protein. PMID- 16375861 TI - The nuclear import of RNA helicase A is mediated by importin-alpha3. AB - RNA helicase A (RHA), an ATPase/helicase, regulates the gene expression at various steps including transcriptional activation and RNA processing. RHA is known to shuttle between the nucleus and cytoplasm. We identified the nuclear localization signal (NLS) of RHA and analyzed the nuclear import mechanisms. The NLS of RHA (RHA-NLS) consisting of 19 amino acid residues is highly conserved through species and does not have the consensus classical NLS. In vitro nuclear import assays revealed that the nuclear import of RHA was Ran-dependent and mediated with the classical importin-alpha/beta-dependent pathway. The binding assay indicated that the basic residues in RHA-NLS were used for interaction with importin-alpha. Furthermore, the nuclear import of RHA-NLS was supported by importin-alpha1 and preferentially importin-alpha3. Our results indicate that the nuclear import of RHA is mediated by the importin-alpha3/importin-beta-dependent pathway and suggest that the specificity for importin may regulate the functions of cargo proteins. PMID- 16375862 TI - Extremely low penetrance of deafness associated with the mitochondrial 12S rRNA mutation in 16 Chinese families: implication for early detection and prevention of deafness. AB - Mutations in mitochondrial DNA (mtDNA) have been found to be associated with sensorineural hearing loss. We report here the clinical, genetic, and molecular characterization of 16 Chinese pedigrees (a total of 246 matrilineal relatives) with aminoglycoside-induced impairment. Clinical evaluation revealed the variable phenotype of hearing impairment including audiometric configuration in these subjects, although these subjects share some common features: being bilateral and sensorineural hearing impairment. Strikingly, these Chinese pedigrees exhibited extremely low penetrance of hearing loss, ranging from 4% to 18%, with an average of 8%. In particular, nineteen of 246 matrilineal relatives in these pedigrees had aminoglycoside-induced hearing loss. Mutational analysis of the mtDNA in these pedigrees showed the presence of homoplasmic 12S rRNA A1555G mutation, which has been associated with hearing impairment in many families worldwide. The extremely low penetrance of hearing loss in these Chinese families carrying the A1555G mutation strongly supports the notion that the A1555G mutation itself is not sufficient to produce the clinical phenotype. Children carrying the A1555G mutation are susceptible to the exposure of aminoglycosides, thereby inducing or worsening hearing impairment, as in the case of these Chinese families. Using those genetic and molecular approaches, we are able to diagnose whether children carry the ototoxic mtDNA mutation. Therefore, these data have been providing valuable information and technology to predict which individuals are at risk for ototoxicity, to improve the safety of aminoglycoside therapy, and eventually to decrease the incidence of deafness. PMID- 16375863 TI - Immune responses to an encapsulated allogeneic islet beta-cell line in diabetic NOD mice. AB - Our goal is to develop effective islet grafts for treating type 1 diabetes. Since human islets are scarce, we evaluated the efficacy of a microencapsulated insulin secreting conditionally transformed allogeneic beta-cell line (betaTC-tet) in non obese diabetic mice treated with tetracycline to inhibit cell growth. Relatively low serum levels of tetracycline controlled proliferation of betaTC-tet cells without inhibiting effective control of hyperglycemia in recipients. There was no significant host cellular reaction to the allografts or host cell adherence to microcapsules, and host cytokine levels were similar to those of sham-operated controls. We conclude that encapsulated allogeneic beta-cell lines may be clinically relevant, because they effectively restore euglycemia and do not elicit a strong cellular immune response following transplantation. To our knowledge, this is the first extensive characterization of the kinetics of host cellular and cytokine responses to an encapsulated islet cell line in an animal model of type 1 diabetes. PMID- 16375864 TI - PI3K-Akt inactivation induced CHOP expression in endoplasmic reticulum-stressed cells. AB - Stress signals that impair the function of the endoplasmic reticulum (ER) can lead to an accumulation of unfolded proteins in the ER causing cell death. Recent studies have indicated that ER stress contributes to several diseases such as neurodegenerative disorders or diabetes. In the present study, we found that Akt down-regulation is important for inducing CHOP expression, an ER stress-induced transcription factor. Treatment with tunicamycin or thapsigargin, ER stress inducers, caused dephosphorylation of Akt from 12 to 24 h and induced cell death. Interestingly, treatment with a PI3K inhibitor alone induced CHOP expression and caused cell death. However, a MEK1 inhibitor induced neither CHOP expression nor cell death. These results indicate that the inactivation of Akt by ER stress induces CHOP expression and causes cell death. Therefore, Akt plays an important role in ER stressed condition and may have important implications for understanding ER stress-related diseases. PMID- 16375865 TI - Autocatalytic tyrosine nitration of prostaglandin endoperoxide synthase-2 in LPS stimulated RAW 264.7 macrophages. AB - In the literature, biological tyrosine nitrations have been reported to depend not only on peroxynitrite but also on nitrite/hydrogen peroxide linked to catalysis by myeloperoxidase. In endotoxin-stimulated RAW 264.7 macrophages, we have detected a major nitrotyrosine positive protein band around 72 kDa and identified it as prostaglandin endoperoxide synthase-2 (PGHS-2). Isolated PGHS-2 in absence of its substrate arachidonate was not only tyrosine-nitrated with peroxynitrite, but also with nitrite/hydrogen peroxide in complete absence of myeloperoxidase. Our data favor an autocatalytic activation of nitrite by PGHS-2 with a subsequent nitration of the essential tyrosine residue in the cyclooxygenase domain. Under inflammatory conditions, nitrite formed via NO synthase-2 may therefore act as an endogenous regulator for PGHS-2 in stimulated macrophages. Nitration of PGHS-2 by the autocatalytic activation of nitrite further depends on the intracellular concentration of arachidonate since arachidonate reacted competitively with nitrite and could prevent PGHS-2 from nitration when excessively present. PMID- 16375866 TI - Diethyl pyrocarbonate, a histidine-modifying agent, directly stimulates activity of ATP-sensitive potassium channels in pituitary GH(3) cells. AB - The ATP-sensitive K(+) (K(ATP)) channels are composed of sulfonylurea receptor and inwardly rectifying K(+) channel (Kir6.2) subunit. These channels are regulated by intracellular ADP/ATP ratio and play a role in cellular metabolism. Diethyl pyrocarbonate (DEPC), a histidine-specific alkylating reagent, is known to modify the histidine residues of the structure of proteins. The objective of this study was to determine whether DEPC modifies K(ATP)-channel activity in pituitary GH(3) cells. Steady-state fluctuation analyses of macroscopic K(+) current at -120 mV produced power spectra that could be fitted with a single Lorentzian curve in these cells. The time constants in the presence of DEPC were increased. Consistent with fluctuation analyses, the mean open time of K(ATP) channels was significantly increased during exposure to DEPC. However, DEPC produced no change in single-channel conductance, despite the ability of this compound to enhance K(ATP)-channel activity in a concentration-dependent manner with an EC(50) value of 16 microM. DEPC-stimulated K(ATP)-channel activity was attenuated by pretreatment with glibenclamide. In current-clamp configuration, DEPC decreased the firing of action potentials in GH(3) cells. A further application of glibenclamide reversed DEPC-induced inhibition of spontaneous action potentials. Intracellullar Ca(2+) measurements revealed the ability of DEPC to decrease Ca(2+) oscillations in GH(3) cells. Simulation studies also demonstrated that the increased conductance of K(ATP)-channels used to mimic DEPC actions reduced the frequency of spontaneous action potentials and fluctuation of intracellular Ca(2+). The results indicate that chemical modification with DEPC enhances K(ATP)-channel activity and influences functional activities of pituitary GH(3) cells. PMID- 16375867 TI - Physiological neuroprotection by melatonin in a 6-hydroxydopamine model of Parkinson's disease. AB - There is considerable evidence that pharmacological doses of the pineal hormone, melatonin, are neuroprotective in diverse models of neurodegeneration including Parkinson's disease. However, there is limited information about the effects of physiological doses of this hormone in similar models. In this study, rats were chronically treated with melatonin via drinking water following partial 6 hydroxydopamine lesioning in the striatum. The two doses of melatonin (0.4 microg/ml and 4.0 microg/ml) were within the reported physiological concentrations present in the serum and cerebrospinal fluid respectively. At 2 weeks after surgery, the higher dose of melatonin significantly attenuated rotational behavior in hemi-parkinsonian rats compared to similarly lesioned animals receiving either vehicle (P < 0.001) or the lower dose of melatonin (P < 0.01). Animals were perfused or sacrificed 10 weeks after commencing melatonin treatment for immunohistochemical or mRNA studies. Animals treated with 4.0 microg/ml melatonin exhibited normal tyrosine hydroxylase (TH) immunoreactivity in the lesioned striatum, whereas little or no TH immunofluorescence was visible in similarly lesioned animals receiving vehicle. In contrast, semiquantitative RT PCR analysis revealed no group differences in TH mRNA, suggesting spontaneous recovery of this transcript as observed previously in partially lesioned animals. There were no significant differences in striatal GDNF mRNA levels between sham and lesioned animals. However, there was a significant (P < 0.01) increase in GDNF mRNA expression in the intact contralateral striata of lesioned animals treated with vehicle. Interestingly, melatonin treatment attenuated this novel compensatory contralateral increase in striatal GDNF expression, presumably due to its neuroprotective effect. These findings support a physiological role for melatonin in protecting against parkinsonian neurodegeneration in the nigrostriatal system. PMID- 16375868 TI - Testing a model for bilingual semantic priming with interlingual homographs: RT and N400 effects. AB - Using a semantic priming paradigm, this study examines the effects of semantic and lexical-orthographic context on reaction times (RTs) and event-related potentials (ERPs) for interlingual homographs. Dutch-English bilinguals performed an English lexical decision task in which homographs like STEM (meaning "voice" in Dutch) were preceded by primes like ROOT or FOOL that were semantically related or unrelated to the English reading of the target word. Homographs were responded to faster following semantically related primes than following unrelated primes. The responses in both conditions were modulated by the relative frequencies of the two readings of the homographs: responses were faster when their English word frequency was high or when their Dutch word frequency was low. In the ERPs, N400 effects, taken to reflect processes of semantic integration, were found for homographs preceded by related primes. Remarkably, the amplitude of the N400 effect was also modulated by word frequency in both the first (Dutch, L1) and the second (English, L2) language. The observed relationship between lexical and semantic variables supports a model for bilingual semantic priming that extends the language nonselective BIA+ model for bilingual word recognition. PMID- 16375869 TI - Diabetes only affects nitric oxide synthase-containing myenteric neurons that do not contain heme oxygenase 2. AB - It has been demonstrated that subpopulations of myenteric neurons are differentially susceptible to the development of neuropathy in diabetes. Within the myenteric plexus are neurons that contain neuronal nitric oxide synthase (nNOS). However, these are not a homogeneous population. Some of the nNOS containing neurons also contain heme oxygenase 2 (HO2). Therefore, the aim of this study was to compare the effects of diabetes on HO2- and nNOS-containing neurons within the myenteric plexus of the rat ileum. Diabetes was induced in male Wistar rats (350-400 g) by a single i.p. injection of buffered streptozotocin (65 mg/kg). After 12 weeks, immunostaining of wholemount preparations of ileum revealed that diabetes induced a significant shift (P < 0.001, chi-squared test for trend) towards increased neuronal cell body size in nNOS-immunoreactive neurons while HO2-immunoreactive neurons remained unaffected. Double-labeling studies revealed that approximately 50% of nNOS-containing neurons also contained HO2 and that the diabetes-induced change in size was confined to nNOS-immunoreactive neurons that did not contain HO2 (P < 0.01). No change in the size distribution occurred in neurons in which nNOS and HO2 were colocalized. Differences in the response of these two subpopulations of nNOS containing neurons to diabetes could occur because they supply different targets within the gastrointestinal tract or indicate that the antioxidant, HO2, protects those nNOS-containing neurons in which it is colocalized, against oxidative stress that occurs in diabetes. PMID- 16375879 TI - Opposing effect of p38 MAP kinase and JNK inhibitors on the development of heart failure in the cardiomyopathic hamster. AB - OBJECTIVE: p38 MAP kinase (p38 MAPK) and c-Jun NH2-terminal kinase (JNK) have been implicated in the pathophysiology of heart failure. We investigated the effects of chronic treatment with p38 MAPK and JNK inhibitors on the development of heart failure in dilated cardiomyopathy (DCM) hamster heart. METHODS AND RESULTS: BIO14.6 hamster hearts showed markedly increased p38 MAPK and JNK activities at 6 weeks of age when there was no significant increase in the area of fibrosis, heart weight/body weight, left ventricular (LV) chamber dilation and LV dysfunction. p38 MAPK and JNK activities were attenuated at 26 weeks of age and abolished at 40 weeks of age in BIO14.6 hamster hearts. BIO14.6 hamsters and the control BIOF1B hamsters were chronically treated (i.p.) with the p38 MAPK inhibitors, SB203580 (1 mg/kg/day) and FR167653 (3 mg/kg/day), or the JNK inhibitor, SP600125 (1 mg/kg/day) or vehicle for 20 weeks starting from 6 weeks of age. Treatment of BIO14.6 hamster hearts with SB203580 and FR167653 reduced the number of TUNEL-positive myocytes, the area of fibrosis and heart weight/body weight associated with a significant decrease of LV dimension and an increase in LV ejection fraction and LV contractility compared to the vehicle-treated counterpart. In contrast, treatment with SP600125 increased the number of TUNEL positive myocytes and the area of interstitial fibrosis associated with aggravation of LV chamber dilation and LV dysfunction. CONCLUSIONS: These results suggest that chronic treatment with p38 MAPK and JNK inhibitors produces opposing effects on the development of heart failure in the DCM hamster heart. PMID- 16375880 TI - Enhancement of antitumor effect of paclitaxel in combination with immunomodulatory Withania somnifera on benzo(a)pyrene induced experimental lung cancer. AB - The current experimental work deals with the immunomodulatory studies on the extract of Withania somnifera (L.) Dunal root powder against benzo(a)pyrene induced lung cancer in male Swiss albino mice. In our previous study, we reported the antioxidant and anticarcinogenic effect of W. somnifera (L.) Dunal along with paclitaxel. Immune dysfunction has been found to be associated with cancer and chemotherapy. Benzo(a)pyrene induced cancer animals were treated with 400mg/kg bodyweight of W. somnifera (L.) Dunal extract for 30 days significantly alters the levels of immunocompetent cells, immune complexes and immunoglobulins. Based on the data, the carcinogen as well as the paclitaxel affects the immune system, the toxic side effects on the immune system is more reversible and more controllable by W. somnifera (L.) Dunal. These results concluded the immunomodulatory activity of W. somnifera (L.) Dunal extract, which is a known immunomodulator in indigenous medicine. PMID- 16375881 TI - Expression of the eukaryotic translation initiation factor 4E (eIF4E) and 4E-BP1 in esophageal cancer. AB - OBJECTIVES: The eukaryotic initiation factor 4E (eIF4E) has been shown to play a key role in cell growth, and several studies have documented an increased expression of eIF4E in a number of solid tumors, including breast, bladder, cervical and head and neck cancers. The aim of this study was to determine the level of expression of eIF4E and eIF4E-binding protein 1 (4E-BP1) in esophageal cancer. DESIGN AND METHODS: Tumor and normal adjacent tissue samples (as controls) were obtained from 99 patients with esophageal cancer. eIF4E and 4E-BP1 levels were determined by using Western blot analysis. eIF4E was purified by affinity chromatography using m(7)GTP-sepharose, and the levels of 4E-BP1 bound to eIF4E were determined. RESULTS: : In cancerous esophageal tissues, eIF4E level was elevated by a mean of 12.59 +/- 1.66-fold, and 4E-BP1 was elevated by a mean of 15.76 +/- 2.45-fold when compared to normal adjacent tissues. Both eIF4E and 4E-BP1 expressions were higher in cancerous compared to noncancerous esophageal tissues. CONCLUSIONS: A higher expression of eIF4E was correlated with advanced stages. The inverse relationship found between the increases in 4E-BP1 levels observed in esophagus tumors and lymph node involvement would support the hypothesis that the overexpression of eIF4E can be involved in tumorigenesis and a possible role of 4E-BP1 as a prognostic factor in esophageal cancer. PMID- 16375882 TI - Spatial knowledge acquisition from direct experience in the environment: individual differences in the development of metric knowledge and the integration of separately learned places. AB - Existing frameworks for explaining spatial knowledge acquisition in a new environment propose either stage-like or continuous development. To examine the spatial microgenesis of individuals, a longitudinal study was conducted. Twenty four college students were individually driven along two routes in a previously unfamiliar neighborhood over 10 weekly sessions. Starting Session 4, they were also driven along a short connecting route. After each session, participants estimated spatial properties of the routes. Some participants' knowledge improved fairly continuously over the sessions, but most participants either manifested accurate metric knowledge from the first session or never manifested accurate metric knowledge. Results are discussed in light of these large individual differences, particularly with respect to the accuracy and development of integrated configurational knowledge. PMID- 16375883 TI - Data mining and clinical data repositories: Insights from a 667,000 patient data set. AB - Clinical repositories containing large amounts of biological, clinical, and administrative data are increasingly becoming available as health care systems integrate patient information for research and utilization objectives. To investigate the potential value of searching these databases for novel insights, we applied a new data mining approach, HealthMiner, to a large cohort of 667,000 inpatient and outpatient digital records from an academic medical system. HealthMiner approaches knowledge discovery using three unsupervised methods: CliniMiner, Predictive Analysis, and Pattern Discovery. The initial results from this study suggest that these approaches have the potential to expand research capabilities through identification of potentially novel clinical disease associations. PMID- 16375884 TI - Effect of osmotic immobilization on refrigerated storage and cryopreservation of sperm from a viviparous fish, the green swordtail Xiphophorus helleri. AB - In this study, refrigerated storage and cryopreservation of sperm from the green swordtail Xiphophorus helleri were investigated. Previous cryopreservation research in this species utilized motile sperm because unlike in most fish species, Xiphophorus sperm can remain continuously motile after collection for a week with refrigerated storage. However, this species reproduces by internal fertilization, and given the significant requirements for motility within the female reproductive tract and potential limitations on sperm energetic capacities, immobilization of sperm prior to insemination could be used to improve fertilization success. Thus, the goal in this study was to use osmotic pressure to inhibit the motility of sperm after collection from X. helleri, and to test the effect of immobilization on refrigerated storage and cryopreservation. The objectives were to: (1) estimate the motility of sperm at different osmotic pressures, and determine an osmotic pressure suitable for immobilization; (2) cryopreserve the immobilized sperm, and estimate the motility after thawing with or without dilution, and (3) compare motility of non immobilized and immobilized sperm after thawing, centrifugation, and washing to remove cryoprotectant. Motility was determined when sperm were suspended in 11 different osmotic pressures (24-500 mOsmol/kg) of Hanks' balanced salt solution (HBSS). Motility was observed between 116 and 425 mOsmol/kg. Sperm were not motile when the osmolality was lower than 116 or higher than 425 mOsmol/kg. Motility of the immobilized (non-motile) sperm could be activated by changing the osmotic pressure to 291-316 mOsmol/kg, and motility of immobilized sperm from hypertonic HBSS (425 mOsmol/kg) was significantly higher than that from hypotonic HBSS (145 mOsmol/kg) after 48 h of storage. At an osmolality of 500 mOsmol/kg, HBSS was used as extender to maintain immobilized sperm during cryopreservation with glycerol as the cryoprotectant. High motility (approximately 55%) was obtained in sperm after thawing when cryopreserved with 10-15% glycerol, and dilution of thawed sperm in fresh HBSS (1:4; V:V) was found to decrease the motility significantly. No difference was found in the motility of thawed sperm cryopreserved with 14% glycerol and extended in 310 and 500 mOsmol/kg HBSS. Washing by centrifugation prolonged the motility of thawed sperm from 24 to 72 h in HBSS at 310 and 500 mOsmol/kg. This study showed that sperm from X. helleri could be immobilized by use of specific osmotic pressures, and that the immobilization did not affect sperm motility after thawing. The immobilization of sperm by osmotic pressure could minimize reduction of the energetic capacities necessary for insemination, traversal, and residence within the female reproductive tract, and fertilization. PMID- 16375885 TI - VEGF-A signaling through Flk-1 is a critical facilitator of early embryonic lung epithelial to endothelial crosstalk and branching morphogenesis. AB - Vascular endothelial growth factor-A (VEGF-A) signaling directs both vasculogenesis and angiogenesis. However, the role of VEGF-A ligand signaling in the regulation of epithelial-mesenchymal interactions during early mouse lung morphogenesis remains incompletely characterized. Fetal liver kinase-1 (Flk-1) is a VEGF cognate receptor (VEGF-R2) expressed in the embryonic lung mesenchyme. VEGF-A, expressed in the epithelium, is a high affinity ligand for Flk-1. We have used both gain and loss of function approaches to investigate the role of this VEGF-A signaling pathway during lung morphogenesis. Herein, we demonstrate that exogenous VEGF 164, one of the 3 isoforms generated by alternative splicing of the Vegf-A gene, stimulates mouse embryonic lung branching morphogenesis in culture and increases the index of proliferation in both epithelium and mesenchyme. In addition, it induces differential gene and protein expression among several key lung morphogenetic genes, including up-regulation of BMP-4 and Sp-c expression as well as an increase in Flk-1-positive mesenchymal cells. Conversely, embryonic lung culture with an antisense oligodeoxynucleotide (ODN) to the Flk-1 receptor led to reduced epithelial branching, decreased epithelial and mesenchymal proliferation index as well as downregulating BMP-4 expression. These results demonstrate that the VEGF pathway is involved in driving epithelial to endothelial crosstalk in embryonic mouse lung morphogenesis. PMID- 16375886 TI - PTHrP induces changes in cell cytoskeleton and E-cadherin and regulates Eph/Ephrin kinases and RhoGTPases in murine secondary trophoblast cells. AB - The differentiation of murine trophoblast giant cells (TGCs) is well characterised at the molecular level and, to some extent, the cellular level. Currently, there is a rudimentary understanding about factors regulating the cellular differentiation of secondary TGCs. Using day 8.5 p.c.-ectoplacental cone (EPC) explant in serum-free culture, we have found parathyroid hormone-related protein (PTHrP) to regulate cellular changes during TGC differentiation. PTHrP greatly stimulated the formation and organisation of actin stress fibres and actin expression in trophoblast outgrowth. This coincided with changing cell shape into a flattened/fibroblastic morphology, suppression of E-cadherin expression, and increased cell spreading in culture. PTHrP also increased the nuclear staining of beta-catenin and, similar to activator protein-2gamma (AP 2gamma), showed microtubule-dependent nuclear localisation in vitro. These cellular and behavioural changes correlated with changes in the expression of RhoGTPases and in both expression and phosphorylation of Eph/Ephrin kinases. The effects of PTHrP on trophoblast cellular differentiation were abolished after blocking its action. In conclusion, PTHrP provides an excellent example of the extrinsic factors that, through their network of activities, plays an important role in cellular differentiation of secondary TGCs. PMID- 16375887 TI - Characterisation of CGRP receptors in human and porcine isolated coronary arteries: evidence for CGRP receptor heterogeneity. AB - This study sets out to characterise calcitonin gene-related peptide (CGRP) receptors in human and porcine isolated proximal and distal coronary arteries using BIBN4096BS. Human (h)-alphaCGRP induced relaxations that were blocked by BIBN4096BS in all arteries studied. In contrast to the other vessels, the Schild plot slope in the human distal coronary artery segments (0.68 +/- 0.07) was significantly less than unity and BIBN4096BS potently blocked these responses (pK(b) (10 nM): 9.29 +/- 0.34, n = 5). In the same preparation, h-alphaCGRP(8-37) behaved as a weak antagonist of h-alphaCGRP-induced relaxations (pK(b) (3 microM): 6.28 +/- 0.17, n = 4), with also a Schild plot slope smaller than unity. The linear agonists, [ethylamide-Cys(2,7)]-h-alphaCGRP ([Cys(Et)(2,7)]-h alphaCGRP) and [acetimidomethyl-Cys(2,7)]-h-alphaCGRP ([Cys(Acm)(2,7)]-h alphaCGRP), had a high potency (pEC(50): 8.21 +/- 0.25 and 7.25 +/- 0.14, respectively), suggesting the presence of CGRP(2) receptors, while the potent blockade by BIBN4096BS (pK(b) (10 nM): 10.13 +/- 0.29 and 9.95 +/- 0.11, respectively) points to the presence of CGRP(1) receptors. Using RT-PCR, mRNAs encoding for the essential components for functional CGRP(1) receptors were demonstrated in both human proximal and distal coronary artery. Further, h alphaCGRP (100 nM) increased cAMP levels, and this was attenuated by BIBN4096BS (1 microM). The above results demonstrate the presence of CGRP(1) receptors in all coronary artery segments investigated, but the human distal coronary artery segments seem to have an additional population of CGRP receptors not complying with the currently classified CGRP(1) or CGRP(2) receptors. PMID- 16375888 TI - Study of the interaction of antiplasmodial strychnine derivatives with the glycine receptor. AB - Strychnos icaja Baill. (Loganiaceae) is a liana found in Central Africa known to be an arrow and ordeal poison but also used by traditional medicine to treat malaria. Recently, many dimeric or trimeric indolomonoterpenic alkaloids with antiplasmodial properties have been isolated from its rootbark. Since these alkaloids are derivatives of strychnine, it was important, in view of their potential use as antimalarial drugs, to assess their possible convulsant strychnine-like properties. In that regard, their interaction with the strychnine sensitive glycine receptor was investigated by whole-cell patch-clamp recordings on glycine-gated currents in mouse spinal cord neurons in culture and by [(3)H]strychnine competition assays on membranes from adult rat spinal cord. These experiments were carried out on sungucine (leading compound of the chemical class) and on the antiplasmodial strychnogucine B (dimeric) and strychnohexamine (trimeric). In comparison with strychnine, all compounds interact with a very poor efficacy and only at concentrations >1 microM with both [(3)H]strychnine binding and glycine-gated currents. Furthermore, the effects of strychnine and protostrychnine, a monomeric alkaloid (without antiplasmodial activity) also isolated from S. icaja and differing from strychnine only by a cycle opening, were compared in the same way. The weak interaction of protostrychnine confirms the importance of the G cycle ring structure in strychnine for its binding to the glycine receptor and its antagonist properties. PMID- 16375889 TI - Gaultherin, a natural salicylate derivative from Gaultheria yunnanensis: towards a better non-steroidal anti-inflammatory drug. AB - One of the major factors limiting the use of non-steroidal anti-inflammatory drugs is gastrointestinal toxicity. Gaultherin, 2-[(6-O-beta-D-Xylopyranosyl-beta D-glucopyranosyl)oxy] benzoic acid methyl ester, a natural salicylate derivative extracted from Gaultheria yunnanensis, has been shown to have analgesic and anti inflammatory effects and lack gastric ulcerogenic effect compared to aspirin in our primary study. The aim of this study was to investigate the mechanism of action of gaultherin, which may rely on its active metabolite, and the mechanism responsible for the non-ulcerogenic property. The results showed that gaultherin (200 mg/kg) significantly inhibited the abdominal contractions in the acetic acid induced writhing test in mice. The anti-inflammatory effect of gaultherin was demonstrated in the croton oil-induced ear edema model in mice. The results showed that gaultherin and equimolar dose of aspirin produced comparable inhibitory effects. The study of the metabolism characters of gaultherin in mice and rats indicated that gaultherin could be metabolically converted to salicylate, which produced the pharmacological effects, and provided effective concentrations for an extended period. In vitro metabolism experiment showed that gaultherin was metabolized by beta-glycosidase produced by human intestinal bacteria and esterases in intestine, blood and liver successively to release salicylate finally. The study suggested gaultherin did not cause gastric ulcer for the reason that it released salicylate in intestine slowly, not in stomach and it left the cyclooxygenase-1 unaffected, which was the source of cytoprotective prostaglandins in gastric epithelium. PMID- 16375890 TI - Synergistic affective analgesic interaction between delta-9-tetrahydrocannabinol and morphine. AB - Evidence for an analgesic interaction between delta-9-tetrahydrocannabinol (Delta(9)-THC) and morphine was sought using an experimental pain model applied to normal volunteers. The study incorporated a double blinded, four treatment, four period, four sequence, crossover design. Subjects received Delta(9)-THC 5 mg orally or placebo and 90 min later morphine 0.02 mg/kg intravenously or placebo. Fifteen minutes later subjects rated the pain associated with the application of thermal stimuli to skin using two visual analog scales, one for the sensory and one for the affective aspects of pain. Among sensory responses, neither morphine nor Delta(9)-THC had a significant effect at the doses used, and there was no significant interaction between the two. Among affective responses, although neither morphine nor Delta(9)-THC had a significant effect, there was a positive analgesic interaction between the two (p = 0.012), indicating that the combination had a synergistic affective analgesic effect. The surprisingly limited reported experimental experience in humans does not support a role for Delta(9)-THC as an analgesic or as an adjunct to cannabinoid analgesia, except for our finding of synergy limited to the affective component of pain. Comparison of our results with those of others suggests that extrapolation from experimental pain models to the clinic is not likely to be a straight-forward process. Future studies of Delta(9)-THC or other cannabinoids in combination with opiates should focus upon clinical rather than experimental pain. PMID- 16375891 TI - Broadening a classic clinical triad: The hypokinetic motor disorder of normal pressure hydrocephalus also affects the hand. AB - The clinical spectrum of normal pressure hydrocephalus is thought to comprise the triad of hypokinetic gait disorder, dementia and urinary incontinence. In contrast, motor abnormalities involving the upper limbs in normal pressure hydrocephalus have not yet received a great deal of attention. The present study was designed to quantitatively assess grasping movements in normal pressure hydrocephalus and to compare the performance with that in Parkinson's disease. Eight subjects with normal pressure hydrocephalus, eight subjects with Parkinson's disease and eight healthy control subjects grasped to lift an instrumented object. The built-up of fingertip forces during the early phase and the kinematics of the lifting movement during the late phase of the grip-lift synergy were slower for patients compared to healthy controls. Patients generated abnormally high fingertip forces when lifting and holding the object stationary. The slowness of the grip-lift synergy and the force overshoot was similar for both patient groups. Our data demonstrate that the hypokinetic motor deficit in normal pressure hydrocephalus also involves the hand, and that the pattern of deficits shares several features of those found in Parkinson's disease. PMID- 16375892 TI - Motor complications in Parkinson's disease and the clinical significance of rotational behavior in the rat: have we wasted our time? PMID- 16375893 TI - Neospora caninum: antibodies directed against tachyzoite surface protein NcSRS2 inhibit parasite attachment and invasion of placental trophoblasts in vitro. AB - Polyclonal and monoclonal antibodies to native Neospora caninum tachyzoite surface protein NcSRS2 were generated and tested in vitro for their ability to neutralize tachyzoite attachment to and invasion of host cells. Host cells included Vero cells and a newly cloned, immortalized ovine trophoblast cell line obtained from primary cultures of ovine placenta. The ovine trophoblasts had morphology consistent with fetal trophoblasts and expressed mRNA for interferon tau, a marker for trophoblasts. Native NcSRS2 was used to immunize mice to obtain monospecific anti-NcSRS2 polyclonal serum and anti-NcSRS2 monoclonal antibodies. Compared to irrelevant antibodies, monospecific anti-NcSRS2 serum and two anti NcSRS2 monoclonal antibodies, 100.2.4.4 and 119.4.9.10, significantly blocked invasion of tachyzoites into both trophoblasts and Vero cells. Parasite attachment, assessed by IFA, was significantly reduced by anti-NcSRS2 mAb 100.2.4.4 and monospecific serum. The findings provide rationale to investigate a role for antibodies to NcSRS2 in prevention of N. caninum transplacental transmission in vivo. PMID- 16375894 TI - Pbcrk-1, the Plasmodium berghei orthologue of P. falciparum cdc-2 related kinase 1 (Pfcrk-1), is essential for completion of the intraerythrocytic asexual cycle. AB - The molecular mechanisms underlying gametocytogenesis in malaria parasites are not understood. Plasmodium falciparum cdc2-related kinase 1 (pfcrk-1), a gene that is expressed predominantly in gametocytes, bears homology to the PITSLRE subfamily of cyclin-dependent kinases and has been hypothesized to function as a negative regulator of the cell cycle. We attempted to knock-out pbcrk-1, the P. berghei orthologue of pfcrk-1, but were unable to recover P. berghei parasites with a disrupted pbcrk-1 locus. In contrast, an integration event at this locus that did not result in a loss-of-function of the pbcrk-1 gene was readily observed. This strongly suggests that a functional pbcrk-1 gene product is essential to intraerythrocytic asexual multiplication. PMID- 16375895 TI - Trypanosoma cruzi: Mutations in the 3' untranslated region of calmodulin gene are specific for lineages T. cruzi I, T. cruzi II, and the Zymodeme III isolates. AB - Populations of Trypanosoma cruzi can be clustered in two main phylogenetic lineages, T. cruzi I and T. cruzi II and a third group denominated Zymodeme III (ZIII) has been described. Using 23 isolates representing the two major T. cruzi groups and the Zymodeme III, the 3' untranslated region (3'UTR) of the calmodulin gene was analyzed. Several mutations located on a 330 bp segment of this 3'UTR were observed, among which three important insertion/deletion events, namely (1) a dinucleotide AG present only in ZIII isolates; (2) a 13 bases purine block missing only in ZIII; and (3) a five base GT block in T. cruzi II. Minimum free energy dot plots show that T. cruzi I and T. cruzi ZIII exhibit similar patterns of optimal and sub-optimal folding of this segment. These mutations in 3'UTR of calmodulin raise the possibility that T. cruzi I and ZIII group are sharing common functional routes. PMID- 16375896 TI - ABC transporter architecture and regulatory roles of accessory domains. AB - We present an overview of the architecture of ATP-binding cassette (ABC) transporters and dissect the systems in core and accessory domains. The ABC transporter core is formed by the transmembrane domains (TMDs) and the nucleotide binding domains (NBDs) that constitute the actual translocator. The accessory domains include the substrate-binding proteins, that function as high affinity receptors in ABC type uptake systems, and regulatory or catalytic domains that can be fused to either the TMDs or NBDs. The regulatory domains add unique functions to the transporters allowing the systems to act as channel conductance regulators, osmosensors/regulators, and assemble into macromolecular complexes with specific properties. PMID- 16375897 TI - The multidrug resistance-associated protein (MRP/ABCC) subfamily of ATP-binding cassette transporters in plants. AB - In many different plant species, genes belonging to the multidrug resistance associated protein (MRP, ABCC) subfamily of ABC transporters have been identified. Following the discovery of vacuolar transport systems for xenobiotic or plant-produced conjugated organic anions, plant MRPs were originally proposed to be primarily involved in the vacuolar sequestration of potentially toxic metabolites. Indeed, heterologous expression of different Arabidopsis MRPs in yeast demonstrates their activity as ATP-driven pumps for structurally diverse substrates. Recent analysis of protein-protein interactions and the characterization of knockout mutants in Arabidopsis suggests that apart from transport functions plant MRPs play additional roles including the control of plant transpiration through the stomata. Here, we review and discuss the diverse functions of plant MRP-type ABC transporters and present an organ-related and developmental analysis of the expression of Arabidopsis MRPs using the publicly available full-genome chip data. PMID- 16375898 TI - Identification of an actin-binding site in p47phox an organizer protein of NADPH oxidase. AB - Actin has been reported to enhance the superoxide-generating activity of neutrophil NADPH oxidase in a cell-free system and to interact with p47phox, a regulatory subunit of the oxidase. In the present study, we searched for an actin binding site in p47phox by far-western blotting and blot-binding assays using truncated forms of p47phox. The amino-acid sequence 319-337 was identified as an actin-binding site, and a synthetic peptide of this sequence bound to actin. The sequence shows no homology to other known actin-binding motifs. It is located in the autoinhibitory region of p47phox and includes Ser-328, a phosphorylation site essential for unmasking. Although a phosphorylation-mimetic p47phox mutant bound to actin with a lower affinity than the wild type, the same mutant interacted with filamentous actin more efficiently than the wild type. A mutant peptide p47phox (319-337, Ser328Glu) bound to filamentous actin more tightly than to monomer actin. These results suggest that p47phox moves to cortical actin when it becomes unmasked in the cells. PMID- 16375899 TI - Characterization of a novel Photosystem I-LHCI supercomplex isolated from Chlamydomonas reinhardtii under anaerobic (State II) conditions. AB - A novel supercomplex of Photosystem I (PSI) with light harvesting complex I (LHCI) was isolated from the green alga Chlamydomonas reinhardtii. This novel supercomplex is unique as it is the first stable supercomplex of PSI together with its external antenna. The supercomplex contains 256 chlorophylls per reaction center. The supercomplex was isolated under anaerobic conditions and may represent the State II form of the photosynthetic unit. In contrast to previously reported supercomplexes isolated in State I, which contain only 4 LHC I proteins, this supercomplex contains 10-11 LHC I proteins tightly bound to the PSI core. In contrast to plants, no LHC II is tightly bound to the PSI-LHCI supercomplex in State II. Investigation of the energy transfer from the antenna system to the reaction center core shows that the LHC supercomplexes are tightly coupled to the PSI core, not only structurally but also energetically. The excitation energy transfer kinetics are completely dominated by the fast phase, with a near complete lack of long-lived fluorescence. This tight coupling is in contrast to all reports of energy transfer in PSI-LHCI supercomplexes (in State I), which have so far been described as weakly coupled supercomplexes with low efficiency for excitation energy transfer. These results indicate that there are large and dynamic changes of the PSI-LHCI supercomplex during the acclimation from aerobic (State I) to anaerobic (State II) conditions in Chlamydomonas. PMID- 16375900 TI - Evidence for phosphorylation of rat liver glucose-regulated protein 58, GRP58/ERp57/ER-60, induced by fasting and leptin. AB - Glucose-regulated protein 58 (GRP58)-like immunoreactivity in rat liver obtained in the evening or after fasting underwent an electrophoretic band-shift, which disappeared after phosphatase-treatment. Since mass spectrometric analysis raised a possibility that Ser150 of GRP58 is phosphorylated, an antibody against the phosphoserine150 GRP58 was generated. Immunoreactivity to this antibody was increased in the evening and after fasting. Since GRP58 was shown to interact with signal transducer and activator of transduction 3 (STAT3), a leptin-related protein, the effect of leptin was examined. Immunoreactivity to the anti phosphoGRP58 antibody was markedly elevated after the leptin injection, indicating that Ser150 of GRP58 is phosphorylated after fasting and leptin treatment. PMID- 16375901 TI - Delta and mu opioid receptors from the brain of a urodele amphibian, the rough skinned newt Taricha granulosa: cloning, heterologous expression, and pharmacological characterization. AB - Two full-length cDNAs, encoding delta (delta) and mu (mu) opioid receptors, were cloned from the brain of the rough-skinned newt Taricha granulosa, complementing previous work from our laboratory describing the cloning of newt brain kappa (kappa) and ORL1 opioid receptors. The newt delta receptor shares 82% amino acid sequence identity with a frog delta receptor and lower (68-70%) identity with orthologous receptors cloned from mammals and zebrafish. The newt mu receptor shares 79% sequence identity with a frog mu receptor, 72% identity with mammalian mu receptors, and 66-69% identity with mu receptors cloned from teleost fishes. Membranes isolated from COS-7 cells transiently expressing the newt delta receptor possessed a single, high-affinity (Kd = 2.4 nM) binding site for the nonselective opioid antagonist [3H]naloxone. In competition binding assays, the newt delta receptor displayed highest affinity for Met-enkephalin, relatively low affinity for Leu-enkephalin, beta-endorphin, and [D-penicillamine, D penicillamine] enkephalin (DPDPE) (a delta-selective agonist in mammals), and very low affinity for mu-, kappa-, or ORL1-selective agonists. COS-7 cells expressing the newt mu receptor also possessed a high-affinity (Kd = 0.44 nM) naloxone-binding site that showed highest affinity for beta-endorphin, moderate to-low affinity for Met-enkephalin and Leu-enkephalin and DAMGO (a mu-selective agonist in mammals), and very low affinity for DPDPE and kappa- or ORL1-selective agonists. COS-7 cells expressing either receptor type (delta or mu) showed very high affinity (Kd = 0.1-0.3 nM) for the nonselective opioid antagonist diprenorphine. Taricha granulosa expresses the same four subtypes (delta, mu, kappa, and ORL1) of opioid receptors found in other vertebrate classes, but ligand selectivity appears less stringent in the newt than has been documented in mammals. PMID- 16375902 TI - Differential expression of tuberoinfundibular peptide 38 and glucose-6 phosphatase in tilapia. AB - A new parathyroid hormone (PTH)-like endocrine system has been identified in mammals and fishes consisting of the PTH type-2 receptor (PTH2R) and tuberoinfundibular peptide 39 (TIP39). Although the mammalian PTH2R-TIP39 system is involved in nociception and pituitary regulation, the function(s) of this system in fishes is undetermined. Using degenerate primers based on conserved zebrafish and fugu TIP39 nucleotide sequences, 3'-RACE reactions isolated the coding region of a putative TIP38 cDNA in the Nile tilapia (Oreochromis niloticus). Tilapia-specific primers were subsequently used in 5'-RACE reactions to isolate the remaining portion of the transcript. The cDNA encoding O. niloticus TIP (OnTIP38) was determined to yield a 38 amino acid secreted hormone. A second tilapia TIP38 cDNA was isolated from the euryhaline Mozambique tilapia (OmTIP38). Except for the 39th residue, both tilapia cDNA sequences showed significant identity to human, bovine, murine, fugu, and zebrafish TIP39. To determine the tissue-specific expression of OnTIP38 and OmTIP38, real-time quantitative RT-PCR (rQRT-PCR) was performed on skin, gill, kidney, testis, heart, and brain. In freshwater (FW)-acclimated Nile tilapia, OnTIP38 showed highest levels of expression in kidney and lowest levels in skin and gill. In Mozambique tilapia tissues, expression of OmTIP38 and G6Pase (glucose-6 phosphatase) were higher in salt water (SW)-acclimated fish than in FW-acclimated fish. G6Pase expression, and not OmTIP38, showed significant differences among various tissues in FW- and SW-acclimated fish. Results of the present study clearly indicate that the TIP38/39-PTH2R system shows considerable conservation in sequence identity and tissue-specific expression in mammals and fishes. PMID- 16375903 TI - Haploinsufficiency of the arginine-vasopressin gene is associated with poor spatial working memory performance in rats. AB - Behavioral pharmacological studies have implicated a role for the neurophysin arginine-vasopressin in learning and memory. Vasopressin, and its analogues, can produce either improvements or impairments in mnemonic functions, effects that depend upon the agent administered, the memory process measured and the task employed. As recent data have implicated vasopressin in regulating the cognitive functions of the prefrontal cortex, we sought to determine whether changes in vasopressinergic tone would affect a form of memory that is dependent upon this brain region. To that end, we used a genetic approach to examine how haploinsufficiency of the vasopressin gene affects working memory performance. Specifically, we tested a naturally occurring null-mutant rat on an operant delayed-non-match-to-position task. Male and female heterozygous and wild-type rats were trained to perform this working memory task, and the effects of varying the delay across which they had to maintain task information were systematically varied. Although vasopressin-deficient rats omitted fewer trials and completed trials more quickly, they exhibited delay-dependent deficits of choice accuracy. The genotype effects were not modified by sex. Collectively, these data indicate that even partial vasopressin deficiency can trigger deficits of spatial working memory performance and add to the growing body of results supporting a regulatory control of neocortical-dependent cognitive functions by this neurohormone. PMID- 16375904 TI - A prospective, population-based, follow-up study of mild traumatic brain injury in children. AB - OBJECTIVE: Outline the annual incidence of mild traumatic brain injury in children, aged 0-17 years, using the American Congress of Rehabilitation Medicine classification and record the presence of new symptoms up to six months after injury. SETTINGS AND METHODS: The study population comprised all the children in the age group of 0-17 years (a total of 40,984) in an administrative district in south-western Sweden. All the individuals registered in the Brain Injury Register, during the periods 1 January to 30 June 1999 and 1 April to 30 September 2000, fulfilling the inclusion criteria defined by American Congress of Rehabilitation Medicine, were included. New symptoms occurring three to six months after the injury were recorded using a mailed 21-item questionnaire. RESULTS: 192 children fulfilled the criteria, yielding an annual incidence of 468/100,000 (95% CI 402-535/100,000). Boys accounted for 57 per cent and fall injuries accounted for 61 per cent of the external causes. New symptoms were reported by 24 per cent of those who answered the questionnaire. CONCLUSION: This study revealed that the annual incidence of mild traumatic brain injury in children was almost as high as that among adults in the same area and population. PMID- 16375905 TI - Compression sutures for uterine atony and hemorrhage following cesarean delivery. PMID- 16375906 TI - Bacterial contamination of women in labor supervised by traditional birth attendants. PMID- 16375907 TI - Biosynthesis and uptake of thiamine (vitamin B1) in bloodstream form Trypanosoma brucei brucei and interference of the vitamin with melarsen oxide activity. AB - Bloodstream forms of Trypanosoma brucei brucei were cultivated in the presence and absence of thiamine (vitamin B1) and pyridoxine (vitamin B6). The vitamins do not change growth behaviour, indicating that Trypanosoma brucei is prototrophic for the two vitamins even though in silico no bona-fide thiamine-biosynthetic genes could be identified in the T. brucei genome. Intracellularly, thiamine is mainly present in its diphosphate form. We were unable to detect significant uptake of [3H]thiamine and structural thiamine analogues such as pyrithiamine, oxithiamine and amprolium were not toxic for the bloodstream forms of T. brucei, indicating that the organism does not have an efficient uptake system for thiamine and its analogues. We have previously shown that, in the fission yeast Saccharomyces pombe, the toxicity of melarsen oxide, the pharmacologically active derivative of the frontline sleeping sickness drug melarsoprol, is abolished by thiamine and the drug is taken up by a thiamine-regulated membrane protein which is responsible for the utilization of thiamine. We show here that thiamine also has weak effects on melarsen oxide-induced growth inhibition and lysis in T. brucei. These effects were consistent with a low affinity of thiamine for the P2 adenosine transporter that is responsible for uptake of melaminophenyl arsenicals in African trypanosomes. PMID- 16375908 TI - Simvastatin induces heat shock factor 1 in vascular endothelial cells. AB - Statins not only reduce serum cholesterol but they also improve vascular endothelial function independent of their lipid-lowering effects. However, except for the mechanism of nitric oxide induction via calveolin, the physiologic basis for the pleiotropic effect of statins remains unknown. In the present study, we investigated the relationship between the effects of statins on vascular endothelial cell function and heat shock proteins. We found that, in vascular endothelial cells, simvastatin increased the steady-state levels of heat shock proteins 90 and 70, and heme oxygenase-1 and caused the nuclear translocation of heat shock factor 1. A decoy oligonucleotide encoding the heat shock element inhibited statin-induced expression of heat shock protein 70, endothelial nitric oxide synthase, and thrombomodulin. This decoy oligonucleotide also inhibited the ability of statin to reduce endothelin-1 and plasminogen activator inhibitor-1 expression. These results indicate that statins improve vascular endothelial function via heat shock factor 1, which may contribute to their ability to improve cardiovascular disease. PMID- 16375909 TI - Cohesive finite element modeling of age-related toughness loss in human cortical bone. AB - Although the age-related loss of bone quality has been implicated in bone fragility, a mechanistic understanding of the relationship is necessary for developing diagnostic and treatment modalities in the elderly population at risk of fracture. In this study, a finite element based cohesive zone model is developed and applied to human cortical bone in order to capture the experimentally shown rising crack growth behavior and age-related loss of bone toughness. The cohesive model developed here is based on a traction-crack opening displacement relationship representing the fracture processes in the vicinity of a propagating crack. The traction-displacement curve, defining the cohesive model, is composed of ascending and descending branches that incorporate material softening and nonlinearity. The results obtained indicate that, in contrast to initiation toughness, the finite element simulations of crack growth in compact tension (CT) specimens successfully capture the rising R-curve (propagation toughness) behavior and the age-related loss of bone toughness. In close correspondence with the experimentally observed decrease of 14-15% per decade, the finite element simulation results show a decrease of 13% in the R-curve slope per decade. The success of the simulations is a result of the ability of cohesive models to capture and predict the parameters related to bone fracture by representing the physical processes occurring in the vicinity of a propagating crack. These results illustrate that fracture mechanisms in the process zone control bone toughness and any modification to these would cause age-related toughness loss. PMID- 16375910 TI - Effect of vibration magnitude, vibration spectrum and muscle tension on apparent mass and cross axis transfer functions during whole-body vibration exposure. AB - Twelve seated male subjects were exposed to 15 vibration conditions to investigate the nature and mechanisms of the non-linearity in biomechanical response. Subjects were exposed to three groups of stimuli: Group A comprised three repeats of random vertical vibration at 0.5, 1.0 and 1.5 ms(-2) r.m.s. with subjects sitting in a relaxed upright posture. Group B used the same vibration stimuli as Group A, but with subjects sitting in a 'tense' posture. Group C used vibration where the vibration spectrum was dominated by either low-frequency motion (2-7 Hz), high-frequency motion (7-20 Hz) or a 1.0 ms(-2) r.m.s. sinusoid at the frequency of the second peak in apparent mass (about 10-14 Hz) added to 0.5 ms(-2) r.m.s. random vibration. In the relaxed posture, frequencies of the primary peak in apparent mass decreased with increased vibration magnitude. In the tense posture, the extent of the non-linearity was reduced. For the low frequency dominated stimulus, the primary peak frequency was lower than that for the high-frequency dominated stimulus indicating that the frequency of the primary peak in the apparent mass is dominated by the magnitude of the vibration encompassing the peak. Cross-axis transfer functions showed peaks of about 15-20% and 5% of the magnitudes of the peaks in the apparent mass for x- and y-direction transfer functions, respectively, in the relaxed posture. In the tense posture, cross-axis transfer functions reduced in magnitude with increased vibration, likely indicating a reduced fore-aft pitching of the body with increased tension, supporting the hypothesis that pitching contributes to the non-linearity in apparent mass. PMID- 16375911 TI - Identification of phenylpropanoids in methyl jasmonate treated Brassica rapa leaves using two-dimensional nuclear magnetic resonance spectroscopy. AB - Metabolic analysis showed a clear increase in phenylpropanoid levels in Brassica rapa leaves after treatment with methyl jasmonate. A fraction of phenylpropanoids was prepared by Diaion HP-20 and Sephadex LH-20 column chromatography after MeOH water extraction. Even with these purification steps, isolation of each phenylpropanoid for structure elucidation is not easy due to the low levels in the plants (ca. 0.004%). A mixture was analyzed without further purification using HPLC-electrospray ionization mass spectrometry and NMR spectroscopy. Based on the NMR results including (1)H NMR, J-resolved, correlated spectroscopy (COSY), heteronuclear single quantum coherence (HSQC), and heteronuclear multiple bond correlation (HMBC) spectra, both (1)H and (13)C resonances of the compounds were obtained. Using these NMR data, five phenylpropanoids conjugated with malate were identified: 5-hydroxyferuloyl-, caffeoyl-, coumaroyl-, feruloyl-, and sinapoyl malate. Of the compounds, 5-hydroxyferuloyl malate is a new phenylpropanoid. In addition to the five constitutive phenylpropanoids bearing trans-configuration, their cis forms, which are believed to be artifacts formed in the course of extraction steps, were also identified in the fraction. PMID- 16375912 TI - An evaluation of solid phase microextraction for aliphatic amines using derivatization with 9-fluorenylmethyl chloroformate and liquid chromatography. AB - The reliability of SPME combined with a chemical reaction for the analysis of short-chain aliphatic amines by liquid chromatography has been investigated. Different options to couple SPME and derivatization have been tested and compared: (i) derivatization of the analytes in solution followed by the extraction of the derivatives, (ii) extraction of the analytes and subsequent derivatization by immersing the SPME fibre onto a solution of the reagent, and (iii) extraction/derivatization of the analytes using fibres previously coated with the reagent. Methylamine (MA), dimethylamine (DMA) and trimethylamine (TMA) have been selected as a model of primary, secondary and tertiary amines, respectively. The analytes have been derivatized with the fluorogenic reagent 9 fluorenylmethyl chloroformate (FMOC), and the fibre coating was Carbowax templated resin (CW-TR). The employment of fibres coated with FMOC to extract and derivatize the analytes was the best option, as compared with the other approaches tested the sensitivity was considerably improved. On the basis of these studies, a new procedure for the determination of MA, DMA and TMA in water is presented. To demonstrate the utility of the proposed conditions data on linearity, accuracy, repeatability and sensitivity are given. Results of the determination of the amines in tap, river and waste water are also presented. PMID- 16375913 TI - Adsorption-induced fibronectin aggregation and fibrillogenesis. AB - Fibronectin (Fn), a high molecular weight glycoprotein, is a central element of extracellular matrix architecture that is involved in several fundamental cell processes. In the context of bone biology, little is known about the influence of the mineral surface on fibronectin supramolecular assembly. We investigate fibronectin morphological properties induced by its adsorption onto a model mineral matrix of hydroxyapatite (HA). Fibronectin adsorption onto HA spontaneously induces its aggregation and fibrillation. In some cases, fibronectin fibrils are even found connected into a dense network that is close to the matrix synthesized by cultured cells. Fibronectin adsorption-induced self assembly is a time-dependant process that is sensitive to bulk concentration. The N-terminal domain of the protein, known to be implicated in its self-association, does not significantly inhibit the protein self-assembly while increasing ionic strength in the bulk alters both aggregation and fibrillation. The addition of a non-ionic surfactant during adsorption tends to promote aggregation with respect to fibrillation. Ultimately, fibronectin fibrils appear to be partially structured like amyloid fibrils as shown by thioflavine T staining. Taken together, our results suggest that there might be more than one single organization route involved in fibronectin self-assembly onto hydroxyapatite. The underlying mechanisms are discussed with respect to Fn conformation, Fn/surface and Fn/Fn interactions, and a model of fibronectin fibrillogenesis onto hydroxyapatite is proposed. PMID- 16375914 TI - Single, binary and multi-component adsorption of copper and cadmium from aqueous solutions on Kraft lignin--a biosorbent. AB - A new biosorbent for removing toxic metal ions from water/industrial wastewater has been investigated using by-product lignin from paper production. Lignin was extracted from black liquor waste, characterized and utilized for the removal of copper and cadmium from aqueous solutions in single, binary and multi-component systems. Adsorption studies were conducted at different temperatures, lignin particle sizes, pHs and solid to liquid ratios. All the studies were conducted by a batch method to determine equilibrium and kinetic parameters. The Langmuir and Freundlich isotherm models were applied. The Langmuir model fits best the equilibrium isotherm data. The maximum lignin adsorption capacities at 25 degrees C were 87.05 mg/g (1.37 mmol/g) and 137.14 mg/g (1.22 mmol/g) for Cu(II) and Cd(II), respectively. Adsorption of Cu2+ (68.63 mg/g at 10 degrees C and 94.68 mg/g at 40 degrees C) and Cd2+ (59.58 mg/g at 10 degrees C and 175.36 mg/g at 40 degrees C) increased with an increase in temperature. Copper and cadmium adsorption followed pseudo-second order rate kinetics. From kinetic studies, various rate and thermodynamic parameters such as effective diffusion coefficients, activation energy, and activation entropy were evaluated. Adsorption occurs through a particle diffusion mechanism at temperatures 10 and 25 degrees C while at 40 degrees C it occurs through a film diffusion mechanism. The sorption capacity of black liquor lignin is higher than many other adsorbents/carbons/biosorbents utilized for the removal of Cu(II) and Cd(II) from water/wastewater in single and multi-component systems. PMID- 16375915 TI - Evaluating antigen-specific cytotoxic T lymphocyte responses by a novel mouse granzyme B ELISPOT assay. AB - We have established novel ELISA- and ELISPOT-based assays specific for the detection of a potent cytotoxic mediator, granzyme B (GrB), for the assessment of antigen-specific cytotoxic T lymphocyte responses in mice. The sensitivity and specificity of our assays was demonstrated by ELISA using purified mouse GrB and supernatants and cell lysates of cytotoxic lymphocytes derived from GrB-deficient mice. No reactivity was observed by the GrB ELISA in GrB-deficient cells. The mouse GrB ELISPOT was successfully employed to detect antigen-specific effector cell responses of two CTL clones, producing GrB ELISPOT results that correlated strongly with target cell lysis, as assessed by 51Cr-release. Furthermore, we were able to demonstrate direct correlations between GrB ELISPOT and killing by LCMV gp33-specific effector and memory T cells generated in vivo. Thus, the mouse GrB ELISPOT may be used to detect cytotoxic responses, at the single-cell level, for the functional assessment of antigen-specific CD8+ T cell responses in mouse models of infection. PMID- 16375916 TI - Identification of Paenibacillus larvae to the subspecies level: an obstacle for AFB diagnosis. AB - This study was initially aimed at developing a PCR-test to differentiate between the pathogenic agent of American foulbrood (Paenibacillus larvae subsp. larvae) and powdery-scale disease (P. larvae subsp. pulvifaciens) of the honeybee. The test was based on the "insert of clone 9" (iC9), referring to a cloned 1.9 kB HaeIII fragment that occurs only in the P. larvae subsp. larvae reference strains and possibly correlates with American foulbrood virulence. It was shown that an iC9-based PCR-test discriminates between the BCCM/LMG reference strains of both subspecies. However, the screening of 179 Belgian field strains revealed five isolates that gave no iC9-based amplicon, thus rather resembling to P. larvae subsp. pulvifaciens. In addition, they all produced acid from mannitol, a characteristic previously assigned to the pulvifaciens subspecies. Because the reference strains gave conflicting data, this carbohydrate acidification was not conclusive. Therefore, the exact taxonomic position of the five retained strains was determined by a polyphasic approach using SDS-PAGE, AFLP, and ERIC-based PCR. Four iC9-negative field strains could be identified as P. larvae subsp. larvae; the taxonomic position of the fifth field strain remained ambiguous. The latter was provisionally classified as a subspecies pulvifaciens strain on the basis of SDS-PAGE. The present paper demonstrates the existence of field strains that do not fit well in the subdivision of the species P. larvae into two subspecies. Knowing that only one of both subspecies represents the pathogenic agent of AFB, this is a serious obstacle for the diagnosis of this honeybee disease. PMID- 16375917 TI - Virus-like particles in the ovaries of Microctonus aethiopoides Loan (Hymenoptera: Braconidae): comparison of biotypes from Morocco and Europe. AB - Virus-like particles (MaVLP) have been discovered in the ovarial epithelial cells of the solitary, koinobiont, endoparasitoid, Microctonus aethiopoides Loan (Hymenoptera: Braconidae) introduced to New Zealand originally from Morocco to control the lucerne pest Sitona discoideus Gyllenhal (Coleoptera: Curculionidae). MaVLP have been found in all females examined. It has been suggested, although not demonstrated, that like many other such VLP found in parasitoids, MaVLP might play a role in host immunosuppression. Since another biotype of M. aethiopoides from Ireland has been proposed for introduction to control the white clover pest, Sitona lepidus Gyllenhal, in New Zealand, it was considered that females from this biotype warranted transmission electron microscope examination for VLP. No VLP were observed in ovarian tissues of specimens collected from three different locations in Ireland. Similarly, none were found in M. aethiopoides sourced from France, Wales, and Norway. These observations are discussed in relation to quarantine host specificity tests with the Irish biotype, which found that the host range of the Irish biotype is likely to be less extensive than that of the Moroccan biotype already in New Zealand. PMID- 16375918 TI - Crystal structures of mouse autocrine motility factor in complex with carbohydrate phosphate inhibitors provide insight into structure-activity relationship of the inhibitors. AB - Autocrine motility factor (AMF), a tumor-secreted cytokine, stimulates cell migration in vitro and metastasis in vivo. AMF is identical to the extracellular cytokines neuroleukin and maturation factor and, interestingly, to the intracellular enzyme phosphoglucose isomerase. The cytokine activity of AMF is inhibited by carbohydrate phosphate compounds as they compete for AMF binding with the carbohydrate moiety of the AMF receptor (AMFR), which is a glycosylated seven transmembrane helix protein. Here, we report the first comprehensive high resolution crystal structure analyses of the inhibitor-free form and the eight types of inhibitor (phosphate, erythrose 4-phosphate (E4P), arabinose 5-phosphate (A5P), sorbitol 6-phosphate (S6P), 6-phosphogluconic acid (6PGA), fructose 6 phosphate (F6P), glucose 6-phosphate (G6P), or mannose 6-phosphate (M6P)) complexes of mouse AMF (mAMF). We assayed the inhibitory activities of these inhibitors against the cytokine activity of mAMF. The inhibitory activities of the six-carbon sugars (G6P, F6P, M6P, and 6PGA) were found to be significantly higher than those of the four or five-carbon sugars (E4P or A5P). The inhibitory activities clearly depend on the length of the inhibitor molecules. A structural comparison revealed that a water-mediated hydrogen bond between one end of the inhibitor and a rigid portion of the protein surface in the shorter-chain inhibitor (E4P) complex is replaced by a direct hydrogen bond in the longer-chain inhibitor (6PGA) complex. Thus, to obtain a new compound with higher inhibitory activities against AMF, water molecules at the inhibitor binding site of AMF should be replaced by a functional group of inhibitors in order to introduce direct interactions with the protein surface. The present structure-activity relationship studies will be valuable not only for designing more effective AMF inhibitors but also for studying general protein-inhibitor interactions. PMID- 16375919 TI - Structural basis of the differential stability and receptor specificity of H-2Db in complex with murine versus human beta2-microglobulin. AB - beta(2)-Microglobulin (beta(2)m) is non-covalently linked to the major histocompatibility complex (MHC) class I heavy chain and interacts with CD8 and Ly49 receptors. Murine MHC class I heavy chains can bind human beta(2)m (hbeta(2)m) and peptide, and such hybrid molecules are often used in structural and functional studies. The replacement of mouse beta(2)m (mbeta(2)m) with hbeta(2)m has several functional consequences for MHC class I complex stability and specificity, but the structural basis for this is presently unknown. To investigate the impact of species-specific beta(2)m subunits on MHC class I conformation, we provide a crystallographic comparison of H-2D(b) in complex with LCMV-derived gp33 peptide and either hbeta(2)m or mbeta(2)m. The conformation of the gp33 peptide is not affected by the beta(2)m species. Comparison of the interface between beta(2)m and the alpha(1)alpha(2) domains of the heavy chain in these two crystal structures reveals a marked increase in both polarity and number of hydrogen bonds between hbeta(2)m and the alpha(1)alpha(2) domains of H 2D(b). We propose that the positioning of two hydrogen bond rich regions at the hbeta(2)m/alpha(1)alpha(2) interface plays a central role in the increased overall stability and peptide exchange capacity in the H-2D(b)/hbeta(2)m complex. These two regions act as bridges, holding and stabilizing the underside of the alpha(1) and alpha(2) helices, enabling a prolonged peptide-receptive conformation of the peptide binding cleft. Furthermore, analysis of H-2D(b) in complex with either mbeta(2)m or hbeta(2)m provides a structural explanation for the differential binding of H-2D(b)/hbeta(2)m to both Ly49A and Ly49C. Our comparative structural study emphasizes the importance of beta(2)m residues at positions 3, 6 and 29 for binding to Ly49A and suggests that sterical hindrance by residue K6 on hbeta(2)m impairs the recognition of Ly49C by H 2D(b)/gp33/hbeta(2)m. Finally, comparison of the two H-2D(b) crystal structures implies that the beta(2)m species may affect the strength of TCR recognition by affecting CD8 binding. PMID- 16375920 TI - Cooperative effects of cofilin (ADF) on actin structure suggest allosteric mechanism of cofilin function. AB - Using site-specific fluorescence probes and cross-linking we demonstrated that cofilin (ADF), a key regulator of actin cellular dynamics, weakens longitudinal contacts in F-actin in a cooperative manner. Differential scanning calorimetry detected a dual nature of cofilin effects on F-actin conformation. At sub stoichiometric cofilin to actin ratios, cofilin stabilized sterically and non cooperatively protomers at the points of attachment, and destabilized allosterically and cooperatively protomers in the cofilin-free parts of F-actin. This destabilizing effect had a long range, with one cofilin molecule affecting more than 100 protomers, and concentration-dependent amplitude that reached maximum at about 1:2 molar ratio of cofilin to actin. In contrast to existing models, our results suggest an allosteric mechanism of actin depolymerization by cofilin. We propose that cofilin is less likely to sever actin filaments at the points of attachment as thought previously. Instead, due to its dual structural effect, spontaneous fragmentation occurs most likely in cofilin-free segments of filaments weakened allosterically by nearby cofilin molecules. PMID- 16375921 TI - Structure of the dodecahedral penton particle from human adenovirus type 3. AB - The sub-viral dodecahedral particle of human adenovirus type 3, composed of the viral penton base and fiber proteins, shares an important characteristic of the entire virus: it can attach to cells and penetrate them. Structure determination of the fiberless dodecahedron by cryo-electron microscopy to 9 Angstroms resolution reveals tightly bound pentamer subunits, with only minimal interfaces between penton bases stabilizing the fragile dodecahedron. The internal cavity of the dodecahedron is approximately 80 Angstroms in diameter, and the interior surface is accessible to solvent through perforations of approximately 20 Angstroms diameter between the pentamer towers. We observe weak density beneath pentamers that we attribute to a penton base peptide including residues 38-48. The intact amino-terminal domain appears to interfere with pentamer-pentamer interactions and its absence by mutation or proteolysis is essential for dodecamer assembly. Differences between the 9 Angstroms dodecahedron structure and the adenovirus serotype 2 (Ad2) crystallographic model correlate closely with differences in sequence. The 3D structure of the dodecahedron including fibers at 16 Angstroms resolution reveals extra density on the top of the penton base that can be attributed to the fiber N terminus. The fiber itself exhibits striations that correlate with features of the atomic structure of the partial Ad2 fiber and that represent a repeat motif present in the amino acid sequence. These new observations offer important insights into particle assembly and stability, as well as the practicality of using the dodecahedron in targeted drug delivery. The structural work provides a sound basis for manipulating the properties of this particle and thereby enhancing its value for such therapeutic use. PMID- 16375922 TI - A single mutation induces amyloid aggregation in the alpha-spectrin SH3 domain: analysis of the early stages of fibril formation. AB - The Src-homology region 3 domain of chicken alpha-spectrin (Spc-SH3) is a small two-state folding protein, which has never been described to form amyloid fibrils under any condition investigated so far. We show here that the mutation of asparagine 47 to alanine at the distal loop, which destabilises similarly the native and folding transition states of the domain, induces the formation of amyloid fibrils under mild acid conditions. Amyloid aggregation of the mutant is enhanced by the increase in temperature, protein concentration and NaCl concentration. The early stages of amyloid formation have been monitored as a function of time and temperature using a variety of biophysical methods. Differential scanning calorimetry experiments under conditions of amyloid formation have allowed the identification of different thermal transitions corresponding to conformational and aggregation processes as well as to the high temperature disaggregation and unfolding of the amyloid fibrils. Aggregation is preceded by a rapid conformational change in the monomeric domain involving about 40% of the global unfolding enthalpy, considerable change in secondary structure, large loss of tertiary structure and exposure of hydrophobic patches to the solvent. The conformational change is followed by formation of a majority of oligomeric species with apparent hydrodynamic radius between 2.5 nm and 10nm, depending on temperature, together with the appearance and progressive growth of protofibrillar aggregates. After these early aggregation stages, long and curved fibrils of up to several micrometers start to develop by elongation of the protofibrils. The calorimetric data indicate that the specific enthalpy of fibril disaggregation and unfolding is relatively low, suggesting a low density of interactions within the fibril structure as compared to the native protein and a main entropy contribution to the stability of the amyloid fibrils. PMID- 16375923 TI - Extended flexible linker structures in the complement chimaeric conjugate CR2-Ig by scattering, analytical ultracentrifugation and constrained modelling: implications for function and therapy. AB - Complement receptor 2 (CR2; CD21) is a membrane-bound regulator of complement activation, being comprised of 15 or 16 short complement repeat (SCR) domains. A recombinant glycosylated human CR2 SCR 1-2 domain pair was engineered with the Fc fragment of a mouse IgG1 antibody to create a chimaera CR2-Ig containing the major ligand binding domains. Such a chimaera has therapeutic potential as a complement inhibitor or immune modulator. X-ray and neutron scattering and analytical ultracentrifugation identified its domain structure in solution, and provided a comparison with controversial folded-back crystal structures for deglycosylated CR2 SCR 1-2. The radius of gyration R(G) of CR2-Ig was determined to be 5.39(+/-0.14) nm and 5.29(+/-0.01) nm by X-ray and neutron scattering, respectively. The maximum dimension of CR2-Ig was determined to be 17 nm. The molecular mass of CR2-Ig ranged between 101,000 Da and 107,000 Da as determined by neutron scattering and sedimentation equilibrium, in good agreement with the sequence-derived value of 106,600 Da. Sedimentation velocity gave a sedimentation coefficient of 4.49(+/-0.11) S. Stereochemically complete models for CR2-Ig were constructed from crystal structures for the CR2 SCR 1-2 and mouse IgG1 Fc fragments. The two SCR domains and the Fc fragment were joined by randomised conformational peptides. The analysis of 35,000 possible CR2-Ig models showed that only those models in which the two SCR domains were arranged in an open V shape in random orientations about the Fc fragment accounted for the scattering and sedimentation data. It was not possible to define one single conformational family of Fab-like fragment relative to the Fc fragment. This flexibility is attributed to the relatively long linker sequence and the absence of the antibody light chain from CR2-Ig. The modelling also confirmed that the structure of CR2 SCR 1-2 is more extended in solution than in its crystal structure. PMID- 16375924 TI - The NMR and X-ray structures of the Saccharomyces cerevisiae Vts1 SAM domain define a surface for the recognition of RNA hairpins. AB - The SAM domain of the Saccharomyces cerevisiae post-transcriptional regulator Vts1 has a high affinity towards RNA hairpins containing a CUGGC pentaloop. We present the 1.6 Angstroms X-ray crystal structure of the Vts1 SAM domain in its unliganded state, and the NMR solution structure of this domain in its RNA-bound state. Both structures reveal a canonical five helix SAM domain flanked by additional secondary structural elements at the N and C termini. The two structures are essentially identical, implying that no major structural rearrangements occur upon RNA binding. Amide chemical shift changes map the RNA binding site to a shallow, basic patch at the junction of helix alpha5 and the loop connecting helices alpha1 and alpha2. PMID- 16375925 TI - Contribution of electrostatic interactions, compactness and quaternary structure to protein thermostability: lessons from structural genomics of Thermotoga maritima. AB - Studies of the structural basis of protein thermostability have produced a confusing picture. Small sets of proteins have been analyzed from a variety of thermophilic species, suggesting different structural features as responsible for protein thermostability. Taking advantage of the recent advances in structural genomics, we have compiled a relatively large protein structure dataset, which was constructed very carefully and selectively; that is, the dataset contains only experimentally determined structures of proteins from one specific organism, the hyperthermophilic bacterium Thermotoga maritima, and those of close homologs from mesophilic bacteria. In contrast to the conclusions of previous studies, our analyses show that oligomerization order, hydrogen bonds, and secondary structure play minor roles in adaptation to hyperthermophily in bacteria. On the other hand, the data exhibit very significant increases in the density of salt-bridges and in compactness for proteins from T.maritima. The latter effect can be measured by contact order or solvent accessibility, and network analysis shows a specific increase in highly connected residues in this thermophile. These features account for changes in 96% of the protein pairs studied. Our results provide a clear picture of protein thermostability in one species, and a framework for future studies of thermal adaptation. PMID- 16375926 TI - Cytokine profiles in women with different subtypes of major depressive disorder. AB - BACKGROUND: Depression has been associated with activation of the immune system. Some studies have shown increased levels of pro-inflammatory cytokines in patients with major depressive disorder (MDD), but conflicting results also have been described. METHODS: Forty-six unmedicated women with MDD were classified in subgroups (melancholic vs. non-melancholic; acute vs. chronic; severe vs. moderate, and episodic vs. recurrent presentations) and compared with 41 healthy controls. Evaluations of serum IL-1beta, IL-6, IFN-gamma and cortisol were performed on both groups. Patients were evaluated prior and after antidepressant treatment. RESULTS: The sub-classification of depression did not predict differences in cytokine levels. Patients currently depressed had similar levels of cytokines and cortisol as healthy controls. After remission of the symptoms, patients with MDD evolved with enhancement of cytokine levels, but no differences were observed in cortisol levels. LIMITATIONS: In patient treatment, two different classes of antidepressants were applied. The dexamethasone/CRH test was not performed to evaluate the HPA axis. CONCLUSIONS: Out-patient women diagnosed with MDD exhibited normal levels of both cortisol and cytokines before treatment, yet demonstrated an increase in cytokines after antidepressant treatment. In some patients with MDD, the presence of acute stress due to hospitalization may indeed contribute and justify the usual finding of higher levels in both cortisol and cytokines. PMID- 16375927 TI - Therapeutic effect of paclitaxel and propolis on lipid peroxidation and antioxidant system in 7,12 dimethyl benz(a)anthracene-induced breast cancer in female Sprague Dawley rats. AB - Breast cancer is one of the most common cancers in women of developed and developing countries. The optimum management of which requires a multidisciplinary approach including the use of certain biochemical and molecular markers. The effect of propolis along with paclitaxel on 7,12 dimethyl benz(a)anthracene (DMBA) induced experimental breast cancer was investigated in female Sprague Dawley rats. Female Sprague Dawley rats were divided into five groups of six animals each. Group I served as normal control animal. Group II animals received DMBA (20 mg in 0.5 ml sunflower oil and 0.5 ml of saline) i.p. to develop mammary tumor by the end of 90 days. Group III were breast cancer animals treated with 33 mg paclitaxel/kg body weight (bw) weekly once for 4 weeks. Group IV were breast cancer-bearing animals treated with 50 mg propolis/kg bw for 30 days. Group V were breast cancer-bearing animals treated with both paclitaxel and propolis as mentioned above. Administration of paclitaxel and propolis effectively suppressed breast cancer, which is revealed by the decrease in the extent of lipid peroxidation (LPO) with concomitant increase in the activities of enzymic antioxidants (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)) and non-enzymic antioxidants (reduced glutathione (GSH), Vitamin C and Vitamin E) levels when compared to breast cancer bearing animals treated with either paclitaxel or propolis alone. From our results, we conclude that propolis is a potent antioxidant and, when given in combination with paclitaxel, offers maximum protection against DMBA induced mammary carcinogenesis. PMID- 16375928 TI - Expression of tissue inhibitor of metalloproteinase-4 (TIMP-4) in endometrium and placenta of rhesus monkey (Macaca mulatta) during early pregnancy. AB - The tissue inhibitors of metalloproteinases (TIMPs) are secreted as important regulators of the matrix metalloproteinases (MMPs). TIMP-4 is the most recently characterized member of the TIMPs family. In the present study, we examined the expression and localization of the TIMP-4 transcript and protein in endometrium and placenta of rhesus monkey (Macaca mulatta) on days 12, 18 and 26 of pregnancy using RT-PCR, in situ hybridization, and immunohistochemistry. The fragment of TIMP-4 gene from rhesus monkey uterine samples shared 95% identity with the corresponding region of human homologue. On day 12 of pregnancy, TIMP-4 mRNA was mainly expressed in the glandular and luminal epithelium. On days 18 and 26 of pregnancy, the expression of TIMP-4 mRNA tended to decline in glandular epithelium and there were strong staining in the placental villi. Furthermore, TIMP-4 mRNA was very faint or undetectable in the stromal cells, endothelial cells of arterioles and myometrium at any stages of pregnancy. The results of immunohistochemical analysis were similar to that of its mRNA. These findings indicate that TIMP-4 might play an important role in glandular secretion, endometrial tissue remodeling and invasion of the trophoblast cells by regulating MMPs in a localized manner in the uteri of rhesus monkey during early pregnancy. PMID- 16375929 TI - SLC5A8 (SMCT1)-mediated transport of butyrate forms the basis for the tumor suppressive function of the transporter. AB - The identification of SLC5A8 as a tumor suppressor gene in colorectal cancer marks, for the first time, the association of a plasma membrane transporter with tumor suppressive properties. The subsequent establishment of the functional identity of SLC5A8 as a Na+-coupled transporter for short-chain monocarboxylates provides a mechanism for the tumor suppressive function of the transporter. Butyrate, a substrate for the transporter, is a histone deacetylase inhibitor and protective against colorectal cancer. This fatty acid is produced in the colonic lumen by bacterial fermentation of dietary fiber. SLC5A8 mediates the concentrative entry of butyrate from the lumen into colonocytes. Consequently, the transport function of SLC5A8 has the ability to influence the acetylation status of histones and hence gene expression in colonocytes. The ability of SLC5A8 to deliver butyrate into colonic epithelial cells most likely underlies the tumor suppressive role of this transporter. PMID- 16375930 TI - Trace metal variations in the shells of Ensis siliqua record pollution and environmental conditions in the sea to the west of mainland Britain. AB - Shells of the pod razor shell (Ensis siliqua) from 13 locations around the west coast of mainland Britain have been analysed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for a range of trace metals including Zn, Cd, Pb, U, Ba, Sr and Mg. The trace metal record in these shells is a proxy record for changes in seawater chemistry during the 1990s. Regional variations exist in the median concentrations of the analysed metals. Barium concentrations are related to increased productivity from sewage sludge dumping at sea. Strontium shows a local relationship to salinity, but there is no clear relationship over the study area, instead high Sr is often associated with high Ba, and may reflect ontogenetic factors such as growth rate. Magnesium shows a seasonal variation within individual shells and can be used to calculate sea surface temperatures from groups of shells. Contaminant metals show a clear regional relationship with known sources, thus high Pb and Zn are typically associated with former metal mining areas (e.g. Cardigan Bay, Anglesey), and high Pb, Zn, Cd and U are associated with industrial activity in Liverpool Bay. Anomalies such as the high U in shells from northern Scotland cannot at present be explained. A seasonal variation of Pb is also seen in Cardigan Bay and Liverpool Bay, relating to increased winter fluxes of these metals to the marine environment. The regional distribution of these metals is consistent with known sources of contamination and patterns of seawater migration around the coast of Britain. PMID- 16375931 TI - Benzo[a]pyrene-enhanced mutagenesis by man-made mineral fibres in the lung of lamda-lacI transgenic rats. AB - In an attempt to examine the interaction of man-made mineral fibres with benzo[a]pyrene (B[a]P), homozygous X-lacI transgenic F344 rats were intratracheally treated with rock (stone) wool RWI and glass wool MMVF 10 fibres together with B[a]P. To analyze the induction of gene mutations by fibres and B[a]P in lung, single doses of 1 and 2 mg fibres/animal or multiple doses of 2 mg fibres/animal were administered weekly on 4 consecutive weeks (total dose 8 mg/animal). B[a]P (10 mg/animal) was administered either simultaneously with fibres (for single dose treatment with fibres) or together with the last fiber treatment (for multiple dose treatment with fibres). Animals were scarified 4 weeks after the last treatment. Benzo[a]pyrene administered simultaneously with RW1 fibres exhibited a strong synergistic effect on mutagenicity, the observed mutant frequency (MF) being more than three-fold higher than the net sum of the MF induced after separate administration of both agents. Our data suggest that DNA adducts induced by simultaneous B[a]P and fiber treatment lead to a strong increase in mutatant frequencies. PMID- 16375932 TI - Multidimensional scaling of ferrous sulfate and basic tastes. AB - The status of metallic sensations as a primary or basic taste category is controversial. Ferrous sulfate (FeSO4) has been suggested as a prototypical metallic chemosensory stimulus. At least part of the metallic sensation from FeSO4 arises from a metallic retronasal smell. The quality of this sensation was studied via multidimensional scaling (MDS) of taste similarities, with and without nasal closure to eliminate retronasal olfactory sensations. The metallic stimulus was embedded in a series containing classical "basic taste" stimuli, alum and monosodium glutamate. With olfaction available, the metallic stimulus plotted away from basic tastes and taste mixtures. Scaled ratings of sensory properties related to metallic taste (iron-nail, copper-penny-like, aftertaste) of FeSO4 decreased with nasal closure. Results are consistent with the idea that ferrous sulfate produces a distinctly different sensation from the traditional basic tastes, which includes both olfactory and oral sensations. PMID- 16375933 TI - Sneeze and you could miss it. PMID- 16375934 TI - Centrally administered PD 140.548 N-methyl-D-glucamine prevents the autonomic responses to duodenal pain in sheep. AB - Cholecystokinin (CCK) released in the CNS inhibits the analgesic action of exogenous opioids and may antagonize analgesia resulting from the activation of an endogenous pain inhibitory system. The aim of this study was to analyse the central action of PD 140.548 N-methyl-D-glucamine--a peptide antagonist of a specific peripheral type CCK receptor--on animal behaviour, catecholamines (CA) and cortisol concentration, as well as clinical symptoms of visceral pain induced by duodenal distension (DD). A 5 min distension of the duodenum wall, using a 10 cm long balloon filled with 40 and/or 80 ml of water (DD 40 and/or DD 80) at animal body temperature, produced a significant increase in plasma CA and cortisol levels, an increase in the heart rate, hyperventilation and other clinical symptoms (inhibition of rumen motility, bleating, teeth grinding, prostration, urination, defecation) that may be related to pain, proportionally to the degree of intestinal distension. Intracerebroventricular administration of PD 140.548 at the dose of 1 or/and 2 mg in toto 10 min before applying DD 40 completely blocked the increase in blood plasma cortisol, epinephrine (E), norepinephrine (NE) and dopamine (DA) concentration. It is suggested that the central inhibitory action of CCK antagonist on the cortisol and catecholamine release produced by visceral pain is due to the inhibition of peripheral CCK1 type receptors in the central centrifugal descending pain facilitatory system in sheep perhaps via the hypothalamic-pituitary-adrenal axis. PMID- 16375935 TI - Estrogen induces phospholipase A2 activation through ERK1/2 to mobilize intracellular calcium in MCF-7 cells. AB - The principal secreted estrogen, 17beta-estradiol rapidly activates signaling cascades that regulate important physiological processes including ion transport across membranes, cytosolic pH and cell proliferation. These effects have been extensively studied in the MCF-7 estrogen-responsive human breast carcinoma cell line. Here, we demonstrate that a physiological concentration of 17beta-estradiol caused a rapid, synchronous and transient increase in intracellular calcium concentration in a confluent monolayer of MCF-7 cells 2-3 min after treatment. This response was abolished when cells were pre-incubated with the phospholipase A(2) (PLA(2)) inhibitor quinacrine or with the cyclooxygenase inhibitor indomethacin. The translocation of GFP-cPLA(2)alpha to perinuclear membranes occurred 1-2 min after 17beta-estradiol treatment; this translocation was concurrent with the transient phosphorylation of cPLA(2)alpha at serine residue 505. The phosphorylation and translocation of cPLA(2) were sensitive to inhibition of the extracellular signal regulated kinase (ERK) signaling cascade and occurred simultaneously with a transient activation of ERK. The phosphorylation of cPLA(2) could be stimulated by membrane impermeable 17beta estradiol conjugated to bovine serum albumen and was blocked by an antagonist of the classical estrogen receptor. Here we show, for the first time, that PLA(2) and the eicosanoid biosynthetic pathway are involved in the 17beta-estradiol induced rapid calcium responses of breast cancer cells. PMID- 16375936 TI - Mechanisms of action underlying the antiandrogenic effects of the fungicide prochloraz. AB - The fungicide prochloraz has got multiple mechanisms of action that may influence the demasculinizing and reproductive toxic effects of the compound. In the present study, Wistar rats were dosed perinatally with prochloraz (50 and 150 mg/kg/day) from gestational day (GD) 7 to postnatal day (PND) 16. Caesarian sections were performed on selected dams at GD 21, while others were allowed to give birth to pups that were followed until PND 16. Prochloraz caused mild dysgenesis of the male external genitalia as well as reduced anogenital distance and retention of nipples in male pups. An increased anogenital distance indicated virilization of female pups. Effects on steroidogenesis in male fetuses became evident as decreased testicular and plasma levels of testosterone and increased levels of progesterone. Ex vivo synthesis of both steroid hormones was qualitatively similarly affected by prochloraz. Immunohistochemistry of fetal testes showed increased expression of 17alpha-hydroxylase/17,20-lyase (P450c17) and a reduction in 17beta-hydroxysteroid dehydrogenase (type 10) expression, whereas no changes in expression of genes involved in testicular steroidogenesis were observed. Increased expression of P450c17 mRNA was observed in fetal male adrenals, and the androgen-regulated genes ornithine decarboxylase, prostatic binding protein C3 as well as insulin-like growth factor I mRNA were reduced in ventral prostates PND 16. These results indicate that reduced activity of P450c17 may be a primary cause of the disrupted fetal steroidogenesis and that an altered androgen metabolism may play a role as well. In vitro studies on human adrenocortical carcinoma cells supported the findings in vivo as reduced testosterone and increased progesterone levels were observed. Overall, these results together indicate that prochloraz acts directly on the fetal testis to inhibit steroidogenesis and that this effect is exhibited at protein, and not at genomic, level. PMID- 16375937 TI - Identification of 45-hydroxy-46,47-dinoryessotoxin, 44-oxo-45,46,47 trinoryessotoxin, and 9-methyl-42,43,44,45,46,47,55-heptanor-38-en-41 oxoyessotoxin, and partial characterization of some minor yessotoxins, from Protoceratium reticulatum. AB - Preparative HPLC purification of a side-fraction obtained during purification of 44,55-dihydroxyyessotoxin (6) afforded fractions containing previously unidentified yessotoxin analogues. Careful analysis of these fractions by HPLC UV, LC-MS3, and NMR spectroscopy, revealed the identities of some of these analogues as 45-hydroxy-46,47-dinoryessotoxin (1), 44-oxo-45,46,47 trinoryessotoxin (2) and 9-methyl-42,43,44,45,46,47,55-heptanor-38-en-41 oxoyessotoxin (5). Numerous other analogues were present but could only be characterized by HPLC-UV and LC-MS3 due to their low abundance. The HPLC-UV and LC-MS3 data confirm the presence of large numbers of yessotoxin analogues, some of which may be oxidative degradation products, in extracts of Protoceratium reticulatum. Compound-1 is the first 46,47-dinoryessotoxin to be identified. PMID- 16375938 TI - Wavelet based noise suppression technique and its application to ultrasonic flaw detection. AB - The noise suppression techniques with wavelet transform (WT) are widely used in non-destructive testing and evaluation (NDT&E), especially in ultrasonics. Complete reconstruction theory with hard or soft thresholds, reconstruction technique based on the singularities of noise and signal, matched filter with an impulse response, and optimal frequency-to-bandwidth ratio of wavelet technique have all been used to analyze ultrasonic signals for noise suppression. But a more simple and effective technique has been pursued for decades. This paper develops a new technique using WT for the right purpose. In this work, WT is treated as a band-pass filter whose central frequency and frequency bandwidth (CF&FB) are determined by the spectra distribution of an ultrasonic signal captured from real testing situation. For the purpose of matching their CF&FB well, a technique for evaluating the optimal scale of a daughter wavelet is carried out too. By acting this daughter wavelet as a band-pass filter, we can obtain excellent de-noising results, even when the signal to noise ratio (SNR) is below -18 dB. The performance of the technique has been done by ultrasonic signals with computer generated white noises. Finally, the experimental verification is performed on a pipeline specimen with man-made small flaws with good results obtained. The results show that the technique is more suitable for processing heavy noised ultrasonic signals, and it can also be used in automatic flaw detection. PMID- 16375939 TI - A mutant fusion (F) protein of simian virus 5 induces hemagglutinin-neuraminidase independent syncytium formation despite the internalization of the F protein. AB - The fusion (F) protein of simian virus 5 strain W3A induces syncytium formation independently of coexpression of the hemagglutinin-neuraminidase protein. This property can be transferred to the F protein of strain WR by replacing the leucine at position 22 with the W3A F counterpart, proline. The resulting mutant L22P has a conformation that is distinct from that of the WR F protein. Se-L22P is a cleavage site mutant of L22P that is cleavable only by addition of exogenous trypsin. We showed here that the cell surface-localized L22P was internalized with a t1/2 of 25 min and degraded in the cell, while the WR F protein was not. The cell surface-localized Se-L22P underwent a significant conformational change upon cleavage. Intriguingly, it disappeared from the cell surface due to its internalization, while inducing extensive syncytium formation. These results indicate that L22P may display an internalization signal during the course of fusion induction. PMID- 16375940 TI - Structural and functional analysis of the baculovirus single-stranded DNA-binding protein LEF-3. AB - The single-stranded DNA-binding protein LEF-3 of Autographa californica multinucleocapsid nucleopolyhedrovirus consists of 385 amino acid residues, forms oligomers, and promotes Mg2+-independent unwinding of DNA duplexes and annealing of complementary DNA strands. Partial proteolysis revealed that the DNA-binding domain of LEF-3 is located within a central region (residues 28 to 326) that is relatively resistant to proteolysis. In contrast, the N-terminus (27 residues) and C-terminal portion (59 residues) are not involved in interaction with DNA and are readily accessible to proteolytic digestion. Circular dichroism analyses showed that LEF-3 is a folded protein with an estimated alpha-helix content of more than 40%, but it is structurally unstable and undergoes unfolding in aqueous solutions at temperatures near 50 degrees C. Unfolding eliminated the LEF-3 domains that are resistant to proteolysis and randomized the digestion pattern by trypsin. The structural transition was irreversible and was accompanied by the generation of high molecular weight (MW) complexes. The thermal treatment inhibited DNA-binding and unwinding activity of LEF-3 but markedly stimulated its annealing activity. We propose that the shift in LEF-3 activities resulted from the generation of the high MW protein complexes, that specifically stimulate the annealing of complementary DNA strands by providing multiple DNA-binding sites and bringing into close proximity the interacting strands. The unfolded LEF-3 was active in a strand exchange reaction suggesting that it could be involved in the production of recombination intermediates. PMID- 16375941 TI - Maturation of the viral core enhances the fusion of HIV-1 particles with primary human T cells and monocyte-derived macrophages. AB - HIV-1 infection requires fusion of viral and cellular membranes in a reaction catalyzed by the viral envelope proteins gp120 and gp41. We recently reported that efficient HIV-1 particle fusion with target cells is linked to maturation of the viral core by an activity of the gp41 cytoplasmic domain. Here, we show that maturation enhances the fusion of a variety of recombinant viruses bearing primary and laboratory-adapted Env proteins with primary human CD4+ T cells. Overall, HIV-1 fusion was more dependent on maturation for viruses bearing X4 tropic envelope proteins than for R5-tropic viruses. Fusion of HIV-1 with monocyte-derived macrophages was also dependent on particle maturation. We conclude that the ability to couple fusion to particle maturation is a common feature of HIV-1 Env proteins and may play an important role during HIV-1 replication in vivo. PMID- 16375943 TI - Differences of monkey and human overt attention under natural conditions. AB - Rhesus monkeys are widely used as animal models of human attention. Such research rests upon the assumption that similar mechanisms underlie attention in both species. Here, we directly compare the influence of low-level stimulus features on overt attention in monkeys and humans under natural conditions. We recorded eye-movements in humans and rhesus monkeys during free-viewing of natural images. We find that intrinsic low-level features, such luminance-contrast, texture contrast and saliency-as predicted by a standard model, are elevated at fixation points in the majority of images. These correlative effects are not significantly different between species. However, local image modifications affect both species differently: moderate modifications, which are in the range of natural fluctuations, attract overt attention in monkeys significantly stronger than they do in humans. In addition, humans show a higher inter-individual consistency regarding which locations they fixate than monkeys, in spite of the similarity for intrinsic low-level features. Taken together, these data demonstrate that under natural conditions-low-level stimulus features affect attention in monkeys and humans differently. PMID- 16375942 TI - Protection against rotavirus shedding after intranasal immunization of mice with a chimeric VP6 protein does not require intestinal IgA. AB - Intranasal immunization of mice with chimeric VP6 and the adjuvant LT(R192G) consistently elicits >95% reductions in fecal rotavirus shedding following challenge. To determine the association between mucosal antibody and protection, we immunized BALB/c wt and J chain knockout (Jch-/-) mice with VP6 and either LT(R192G) or cholera toxin (CT). Both strains developed nearly equal levels of serum rotavirus IgG, but Jch-/- mice, which cannot transport dimeric IgA across epithelial cell surfaces, developed >4-fold higher levels of serum rotavirus IgA. Stool rotavirus IgA was present in wt but undetectable in Jch-/- mice. When challenged with rotavirus strain EDIM, reductions in rotavirus shedding were nearly identical in VP6-immunized wt and Jch-/- mice (i.e., 97% and 92%, respectively; P > 0.01). Th1 CD4 T cell responses were also detected in VP6 immunized animals based on high levels of IFN-gamma and IL-2 found after in vitro VP6 stimulation of spleen cells. Therefore, protection induced by intranasal immunization of mice with VP6 and adjuvant does not depend on intestinal rotavirus IgA antibody but appears to be associated with CD4 T cells. PMID- 16375944 TI - Aviation-related respiratory gas disturbances affect dark adaptation: a reappraisal. AB - This study examined the time course of early scotopic threshold sensitivity during dark adaptation under mild to moderate hypoxia, moderate hypocapnia and hyperoxia, measuring detection time displacement relative to normoxia. Cone rod inflection and early rod adaptation were highlighted using progressively dimmer green flash stimuli. Early scotopic sensitivity was significantly delayed by hypoxia and hastened by hypocapnia and hyperoxia. Effects of respiratory disturbance on dark adaptation include temporal shifts of early scotopic sensitivity while human rod photoreceptors appear functionally hypoxic when breathing air at one atmosphere. At night, supplementary oxygen may benefit aircrew visual sensitivity, even at ground level. PMID- 16375945 TI - New object onsets reduce conscious access to unattended targets. AB - Attention to a visual target can affect perception of a subsequent target for half a second, increasing its sensitivity to backward masking (the attentional blink, AB). In 6 studies, we compared the AB when the second target and its mask had a common onset and when the mask appeared after the target. The results indicate that common-onset masks do not produce large ABs even when there is a feature change or an interruption of the mask after the target but do produce a large AB if the location of the mask is changed. The data suggest that new object onsets reduce conscious access to unattended targets. PMID- 16375946 TI - Substrates for phosphorus removal-potential benefits for on-site wastewater treatment? AB - A large number of substrates potential for removal of phosphorus (P) in wastewater has been reviewed. The substrates consist of natural materials, industrial by-products and man-made products. Most substrates have been investigated in batch and column studies in the laboratory; others have also been tested in field trials. The results from these investigations vary, but a few substrates, e.g. wollastonite, slag material and, to some extent, light weight aggregate products, have demonstrated promising properties with regard to P sorption capacity and hydraulics. The problems of normalisation of data are discussed, as well as the substrates potential benefits for on-site wastewater treatment. PMID- 16375947 TI - Second-order kinetic model for the sorption of cadmium onto tree fern: a comparison of linear and non-linear methods. AB - A comparison was made of the linear least-squares method and a trial-and-error non-linear method of the widely used pseudo-second-order kinetic model for the sorption of cadmium onto ground-up tree fern. Four pseudo-second-order kinetic linear equations are discussed. Kinetic parameters obtained from the four kinetic linear equations using the linear method differed but they were the same when using the non-linear method. A type 1 pseudo-second-order linear kinetic model has the highest coefficient of determination. Results show that the non-linear method may be a better way to obtain the desired parameters. PMID- 16375948 TI - Concentration changes of PCDDs, PCDFs, and dioxin-like PCBs in human breast milk samples as shown by a follow-up survey. AB - Breast milk is known to discharge dioxins from the human body. However, to date, no direct comparison has been made of the concentrations of dioxin analogues in breast milk obtained from identical population after successive deliveries. We present here follow-up survey results of PCDDs, PCDFs, and dioxin-like PCBs concentrations in human breast milk samples from 35 women living in Tokyo from whom samples had been obtained and analyzed to determine the corresponding concentrations in the past. The average concentrations of total PCDDs, PCDFs, dioxin-like PCBs were 8.5, 5.5, 11 TEQ pg/g fat for the first samples and 5.4, 4.0, 6.6 TEQ pg/g fat for the second samples, respectively. The degrees of reduction of total PCDDs and total dioxin-like PCBs were higher than that of total PCDFs because 1,2,3,7,8-PeCDD, 2,3,4,7,8-PeCDF and 3,3',4,4',5-PeCB (#126), which were the predominant congeners among PCDDs, PCDFs, and dioxin-like PCBs, respectively, had different degrees of reduction. Moreover we observed a significant increase of the concentrations of PCDDs, PCDFs, and dioxin-like PCBs in samples from three women in this follow-up survey, and the patterns of increased isomers differed among the three samples. It was conjectured that the increase of the concentrations was due mainly to dietary intake between deliveries. It is important for pregnant women to have a balanced diet to mitigate the exposure of infants to these chemicals. PMID- 16375949 TI - The chemistry of conventional and alternative treatment systems for the neutralization of acid mine drainage. AB - The oxidation of pyritic mining waste is a self-perpetuating corrosive process which generates acid mine drainage (AMD) effluent for centuries or longer. The chemical neutralization of these complex, buffered effluents result in unstable, metal-laden sludges, which require disposal to minimize long-term environmental consequences. A variety of passive treatment systems for AMD, developed in the past two decades, combine limestone and organic substrates in constructed wetlands. These systems work well initially but over the longer term fail due to clogging with and the depletion of available organic carbon. However, some ecologically engineered systems, which exploit the activities of acid reducing microbes in the sediment, rely on photosynthesis in the water column as a source of organic matter. The primary productivity in the water column, which also generates some alkalinity, provides electron donors for the microbial reduction processes in the sediment. In its consideration of 'passive' systems, the literature has placed undue emphasis on sulphate reduction; thermodynamical iron reduction is equally important as is the need to prevent iron oxidation. Secondary precipitates of iron play a significant role in sediment-driven biomineralization processes, which affect the anaerobic degradation of organic matter and the stability of the resulting metal sulfides. One such passive system, which utilized a floating root mass as a source of organic carbon, is described. An extensive review of the literature and the chemical and biogeochemical reactions of AMD treatment systems, lead to the conclusion, that sediment based ecological systems offer the greatest potential for the sustainable treatment of AMD. PMID- 16375950 TI - Feasibility study comparing docetaxel-cisplatin versus docetaxel-carboplatin as first-line chemotherapy for ovarian cancer. AB - OBJECTIVE: To determine the feasibility of docetaxel-cisplatin combination therapy compared with docetaxel-carboplatin combination therapy as first-line chemotherapy for patients with ovarian cancer. METHODS: Fifty patients with International Federation of Gynecology and Obstetrics stage Ic-IV ovarian cancer who underwent primary surgery were randomly assigned to receive treatment with docetaxel-cisplatin (n = 23) or docetaxel-carboplatin (n = 27). Docetaxel 70 mg/m2 and cisplatin 60 mg/m2 or carboplatin to an area under the curve of 5 were administered consecutively on Day 1 of a 3-week cycle, for 3 cycles in patients with stage Ic-II cancer and for over 5 cycles in patients with stage III-IV cancer. Patients were evaluated for treatment-related toxicity in each cycle using the National Cancer Institute Common Toxicity Criteria version 2.0. RESULTS: Five patients (2 in the docetaxel-cisplatin arm and 3 in the docetaxel carboplatin arm) discontinued the treatment at the end of the second course of chemotherapy because of apparent disease progression; however, no patients came off the protocol therapy because of treatment-related toxicity. Overall, 103 cycles of docetaxel-cisplatin treatment and 130 cycles of docetaxel-carboplatin treatment were delivered. The major toxicity was neutropenia in both regimens. The total incidence of grades 3 and 4 neutropenia was 83% (19/23) in the docetaxel-cisplatin arm and 96% (26/27) in the docetaxel-carboplatin arm. The incidence of grade 4 neutropenia was significantly lower in the docetaxel cisplatin arm [39% (9/23) versus 74% (20/27)]. CONCLUSION: Docetaxel-cisplatin combination therapy may be feasible as first-line chemotherapy for patients with ovarian cancer. PMID- 16375951 TI - Prospective evaluations of continuous weekly paclitaxel regimen in recurrent platinum-resistant epithelial ovarian cancer. AB - BACKGROUND: Paclitaxel administered on a weekly basis has been reported to possess both anti-angiogenic and apoptotic-inducing effects. We investigated the activity of a weekly continuous paclitaxel regimen in patients with recurrent platinum-resistant ovarian cancer. METHODS: Patients with recurrent ovarian cancer and documented platinum-resistant disease were treated with weekly intravenous paclitaxel (60-80 mg/m(2)) continuously for up to 24 weeks over an 18 month period. Prospective data collection included: information on patients' demographics together with disease- and treatment-related toxicities. Response was evaluated using radiographic and Ca125 criteria. Chi-square tests were used to test for significant associations between categorical variables. Progression free survival and overall survival time from commencement of weekly treatment were estimated using the Kaplan-Meier method. All P values less than 0.05 were considered to be statistically significant. RESULTS: Thirty-four patients were treated on protocol. Five patients (15%) reported grade 3/4 neurotoxicity at the end of 12 weeks. No dose reduction or treatment delay was required. No significant hematologic toxicity was observed. Responses were evaluable in thirty two patients. Complete response was observed in three patients (9%), and another 14 patients showed a partial response (44%). Seven patients (22%) had disease stabilization. The estimated median progression-free survival and overall survival were 6.10 months (95% CI:3.81-8.39) and 10.43 months (95% CI: 8.49 12.38) respectively from the start of the regimen. CONCLUSION: Continuous weekly paclitaxel is a well-tolerated and active regimen in patients with recurrent platinum-resistant ovarian cancer. PMID- 16375952 TI - Good correlation of HPV DNA test between self-collected vaginal and clinician collected cervical samples by the oligonucleotide microarray. AB - OBJECTIVES: To evaluate the efficacy of self-collected vaginal samples for high risk HPV detection by the HPV oligonucleotide microarray method (HPVDNAChip). METHODS: One hundred and eighteen patients with abnormal Pap smears were included. Self-collected vaginal and clinician-collected cervical samples for HPV testing were obtained. The result of the HPV DNA test was compared with the histopathological diagnosis or colposcopic finding. RESULTS: Of the 118 patients, 42 (35.6%) had >or= cervical intraepithelial neoplasia (CIN) III lesions. Using the HPVDNAChip, high-risk types of HPV were detected in 38 of these 42 patients (90.5%) with the self-collected vaginal samples and in 37 of 42 (88.1%) with the clinician-collected cervical samples. The agreement of HPVDNAchip results between self- and clinician-collected samples was very good (kappa = 0.81) with a 93.2% concordance rate. Multiple HPV infections were found in 17 of 88 (19.3%) HPV positive clinician-collected cervical samples. The rate of multiple HPV infection tended to decrease as the degree of pathologic classification increased. CONCLUSION: Using the HPVDNAchip to assay for HPV infection, results from self collected vaginal samples were compatible with those from clinician-collected cervical samples. PMID- 16375953 TI - Effects of caffeine on olfactory and visual learning in the honey bee (Apis mellifera). AB - Although caffeine is known to improve alertness and arousal in humans and other mammals, its impacts on specific behaviours, including complex cognitive processes, remain controversial. We reasoned that the availability of an easily manipulable, but behaviourally complex invertebrate organism with a simpler nervous system would be beneficial to this field of research. We used a popular behavioural model, the honeybee, to evaluate the effects of caffeine on (1) the development of olfactory learning and (2) the performance in complex learning paradigms, including a 'delayed-match-to-sample' task and visual associative learning. To evaluate the efficacy of caffeine treatment, a variety of doses (0.4 400 ng/1 mg of body mass) were applied topically to tethered individuals. Behavioural testing was performed with either tethered or free-flying adult honeybees. We show that caffeine has marked cognitive effects in this species. In young honeybees, it reduces the age at which restrained individuals are able to learn an olfactory associative task, whereas in older, free-flying bees, caffeine improves both motivation and cognitive performance in complex learning tasks. Our results suggest that the honeybee model may be useful in explaining caffeine related behavioural changes not only in this species, but also in mammalian systems. PMID- 16375954 TI - Pharmacotherapy and smoking cessation at a tobacco dependence clinic. AB - BACKGROUND: Tobacco dependence medications are effective, and combinations may offer advantages. This study evaluates abstinence rates among smokers treated in a tobacco specialist clinic with individual and/or group counseling plus combination pharmacotherapy. METHODS: 790 smokers treated at the Tobacco Dependence Clinic in New Jersey from 2001-2003 and contacted 4 weeks after quit date were studied. Patients received medications and behavioral interventions. Abstinence over the previous 7 days was evaluated at 4 weeks and 6 months. Patients lost to 6-month follow-up were considered still smoking. RESULTS: Overall, 36% of patients were abstinent at 6 months (20% who used no medications, 37% using one medication, 37% using 2 medications, 42% using 3 medications, and 42% using 4+ medications) (P = 0.017). 27% still used medications at 6 months, and had higher abstinence rates (65%) than those who stopped their medications (27%) (P < 0.001). Number of medications predicted abstinence at 4 weeks [adjusted odds ratios = 2.30 (95% CI; 1.27-4.18) for 1 medication, 4.78 (2.72 8.40) for 2 medications, 5.83 (2.98-11.40) for 3 medications, and 11.80 (4.10 33.95) for 4+ medications]. Increasing age, increasing level of education, longer time after waking to first cigarette, more than 7 clinical contacts, and more medications used were related to higher abstinence at 6 months. CONCLUSIONS: Smokers attending a specialist tobacco dependence treatment clinic who used more medications and for longer duration had higher abstinence rates. PMID- 16375955 TI - Channels of cigarette distribution, price and tobacco consumption in Italy. AB - BACKGROUND: Limited information is available on the role of smuggling and of perceived influence of cigarette price on tobacco consumption in Italy. To elucidate the issues, we included specific questions in a survey on smoking in Italy. METHODS: Between March and April 2004, we conducted a survey on 3050 individuals aged 15 or over, representative of the general adult Italian population. The questionnaire included the estimate of the role of various channels of cigarette distribution and information on the self-reported perception of influence of prices on cigarette consumption in the young. RESULTS: Among current smokers, 85.6% bought cigarettes from tobacco shops, 7.5% from vending machines, and 6.9% from other channels of distribution, including smuggling, and internet (plus offered cigarettes). Overall, 35.9% of ever smokers (37.9% of males and 32.8% of females) reported that the prices had an intermediate to high influence on cigarette consumption in the young. Younger and less educated smokers were not more prone to report an influence of prices. CONCLUSIONS: These data indicate that in Italy smuggling now covers a limited proportion of cigarette sales, while cigarette prices have a substantial influence on tobacco consumption in the young. PMID- 16375956 TI - Getting children to eat more fruit and vegetables: a systematic review. AB - BACKGROUND: There is growing recognition of the need to increase consumption of currently suboptimal levels of fruit and vegetables by children, given their known beneficial effects for health. There is, however, a need for a synthesis of the evidence on interventions that might achieve this policy goal. METHODS: A systematic review of published and unpublished studies was carried out by searching 14 publication databases and contacting experts in the fields. All papers in eight languages were considered if they described individual- and population-based interventions and promotion programmes that encouraged the consumption of a diet relatively higher in fruit and/or vegetables in free living, not acutely ill children of both genders, with follow-up periods of at least 3 months, measurement of change in intake and a control group. RESULTS: Fifteen studies focusing on children met the criteria for inclusion in the systematic review. None of the studies reviewed had a detrimental effect on fruit and vegetable consumption. Ten studies had a significant effect, ranging from +0.3 to +0.99 servings/day. CONCLUSIONS: More research is needed to examine in more depth, for longer follow-up periods, the effectiveness of interventions promoting fruit and vegetable consumption. The evidence is strongest in favor of multi-component interventions to increase fruit and vegetable consumption in children. PMID- 16375957 TI - Differentiation between pyometra and cystic endometrial hyperplasia/mucometra in bitches by prostaglandin F2alpha metabolite analysis. AB - Bitches with pyometra are potential emergency cases which may be clinically difficult to differentiate from cases of cystic endometrial hyperplasia (CEH) in combination with mucometra. In the present study plasma prostaglandin F(2alpha), as measured by its main metabolite 15-keto-13,14-dihydro-PGF(2alpha) (PG metabolite) concentrations, blood biochemical and hematological parameters were measured in 59 bitches with pyometra, 10 bitches with CEH and nine controls to determine if PG-metabolite could differentiate between the three uterine conditions. Bitches with pyometra had significantly higher plasma levels of PG metabolite than bitches with CEH (P=0.002) and the controls (P=0.002). PG metabolite analysis alone had a high sensitivity (98.3%) and a high specificity (80.0%) for the differentiation of pyometra versus CEH in bitches where fluid in the uterus was diagnosed. When a combination of PG-metabolite and percentage band neutrophils (PBN) was used for differentiation of the two diagnoses, a sensitivity of 100% and specificity of 90.0% was obtained. This means that the combination of PG-metabolite and PBN analysis allows for differentiation between cases of pyometra and CEH. If the PG-metabolite level in a bitch is >or=4,524 pmol l(-1), there is a 99% probability of the diagnosis pyometra versus CEH. Levels of PG-metabolite >or=3,054 pmol l(-1), >or=2,388 pmol l(-1) or>or=1,666 pmol l(-1) indicates a 95%, 90% or 80% probability of pyometra, respectively. At high PG-metabolite levels (above about 3,000 pmol l(-1)), PG-metabolite alone is enough for differentiation of pyometra versus CEH. The results of the present study showed that PG-metabolite analysis is valuable in the diagnosis and prediction of severity of uterine diseases. PMID- 16375958 TI - Fraunhofer diffraction of short-fiber-reinforced composites aligned by an electric field. AB - OBJECTIVES: This study aims to determine if aligned, short-fiber composites are capable of producing Fraunhofer diffraction patterns similar to those of human enamel sections and to validate the mechanism of diffraction by comparing the experimental and theoretical fiber spacings. METHODS: Rectangular composite specimens were filled with short E-glass fibers (120 microm length, 15 microm diameter) to contents of 1, 5, 10, and 25% (vol%). The fibers were oriented perpendicular to the surface normal using an alternating electric field of 0.75 kV/mm. A He-Ne laser was used with thin slices of specimen placed behind a pinhole to produce diffraction patterns. The locations of the diffraction maxima were used to determine the theoretical slit spacings which were compared to the experimental slit spacings determined by microscopy. RESULTS: The specimens produced diffraction patterns analogous to the theoretical Fraunhofer diffraction of light through multiple slits. This was verified by comparing the theoretical spacing of the fibers calculated from the diffraction pattern with the experimental spacing of the fibers determined from the optical micrographs. SIGNIFICANCE: This work has verified that orientation of short-fiber composites using an electric field can yield composites with sufficient order to produce Fraunhofer diffraction patterns that are qualitatively similar to the diffraction patterns of human enamel sections. PMID- 16375959 TI - The effect of carbamide peroxide treatment on metal ion release from dental amalgam. AB - OBJECTIVES: There is concern that hydrogen peroxide generated by tooth bleaching agents may cause enhanced metal ion release (including mercury) from dental amalgam following contact. The aim of this in vitro study was therefore to investigate the effect of a carbamide peroxide (CP) based tooth bleaching gel on metal ion release from dental amalgam. METHODS: Dental amalgam discs were prepared according to the manufacturers' instructions. These were treated with either a 10% carbamide peroxide (CP) gel or a 0% CP gel for 24h. Discs were carefully wiped with cotton wool before immersion in distilled water (20 ml) for 24h at 37 degrees C. Following immersion, water samples were taken for metal ion release determination (Ag, Cu, Hg and Sn) using inductively coupled plasma mass spectrometry methods. The specimens were further evaluated for surface changes using scanning electron microscopy (SEM) and Talysurf surface roughness measurements. RESULTS: The differences in concentration of metal ions released after treatment with the 10% CP gel and a placebo gel treatment were not statistically significant (p>0.05). For example, mercury release following treatment with the 10% CP gel and the 0% CP gel was found to be 1.17(0.5) and 0.57(0.1)microgcm(-2), respectively. Roughness measurements for samples treated with the 10% CP gel and 0% CP gel were 2.23(0.47) and 1.74(0.16)microm, respectively, again showing no significant difference between groups (p>0.05). SEM images of the amalgam surfaces showed no apparent differences between treatments. SIGNIFICANCE: Treatment with a 10% CP gel did not significantly enhance subsequent metal ion release from dental amalgams compared to a control gel, contradicting previously published studies. PMID- 16375960 TI - Surface modification of titanium by etching in concentrated sulfuric acid. AB - OBJECTIVE: The purpose of this study was to characterize the etching behavior of titanium in concentrated sulfuric acid and discuss its application on surface modification of titanium for biological use. METHODS: Commercially pure titanium (cpTi) plate was etched in 48% H2SO4 at RT -90 degrees C for 0.25-8 h. The weight loss was derived from the weight differences before and after etching. The surfaces after etching were characterized by surface roughness, X-ray diffractometry, and scannning electron spectroscopy. The apparent activation energy of the dissolution of cpTi into acid was derived from an Arrhenius plot of the rate of weight loss versus the acid temperature. RESULTS: The surface roughness of cpTi increased with the acid temperature and etching time. The surface roughness was strongly related to the weight loss. The weight loss increased drastically with the acid temperature after an initial period, which shortened with increasing acid temperature. The apparent activation energy for the dissolution of cpTi in H2SO4 was derived as 67.8 kJ/mol. SIGNIFICANCE: This study indicates that etching with concentrated sulfuric acid is an effective way to modify the surface of titanium for biological applications. PMID- 16375961 TI - Two-year clinical evaluation of lithia-disilicate-based all-ceramic crowns and fixed partial dentures. AB - OBJECTIVES: The aim of this study was to evaluate the clinical performance of crowns and fixed partial dentures (FPDs) made with the Empress 2 system over a 2 year period. METHODS: Twenty anterior or posterior all-ceramic (Empress 2) crowns and 20 anterior or posterior, three-unit fixed partial dentures were fabricated for 15 patients. Evaluations of the restorations were performed at baseline and once a year during the 2-year follow-up period. U.S. Public Health Service criteria were used to examine the marginal adaptation, color match, secondary caries and visible fractures in the restorations. Survival rate of the restorations were determined using Kaplan-Meier statistical analysis. RESULTS: U.S. Public Health Service criteria showed 100% Alpha scores concerning recurrent caries for both crowns and FPDs. No crown fractures were observed during the 2 year follow-up, however, 10 (50%) catastrophic failures of FPDs occurred. Five (25%) failures occurred within the 1-year clinical period and the others (25%) within the second year. SIGNIFICANCE: Single unit Empress 2 all-ceramic crowns exhibited a satisfactory clinical performance over 2-year period. Furthermore, the high fracture rate of Empress 2 FPDs limits the usage of Empress 2 for the fabrication of all-ceramic FPD. PMID- 16375962 TI - On material choice and fracture susceptibility of restored teeth: an asymptotic stress analysis approach. AB - OBJECTIVE: The ultimate success or failure of a restored tooth is largely dependent on clinical management. Clinicians may choose from a number of restorative materials, different clinical techniques and cavity preparation procedures. The purpose of this study was to specifically examine aspects of the material choice holding other factors constant. METHODS: The current paper adopts a fundamental result in the linear theory of elasticity on the singular stress distribution in a bi-materials wedge to analyze the fracture susceptibility of different materials used for the restoration of a tooth. RESULTS: Comparable results are reported for amalgam, gold alloys and ceramic materials. It is shown that due to a wide variety of mechanical properties the application of resin based composites could lead to improved or less fracture resistance of the restored tooth. SIGNIFICANCE: This variety in the mechanical properties for resin based composites could be partially responsible for the contradictory evidences reported by different clinical studies. The present work contributes evidence from an analytical model to assist the restorative dentist in selection of an appropriate restorative material and guide the manufacturing companies on the preferred physical properties of newer designed materials. PMID- 16375963 TI - The nucleation and crystallization of fine grained leucite glass-ceramics for dental applications. AB - OBJECTIVES: The aims of the study were to control the nucleation and crystal growth of selected aluminosilicate glass powders, to produce uniform leucite glass-ceramic microstructures consisting of fine (<1000 nm) grained leucite crystals. METHODS: A starting glass composition of wt%; 64.2% SiO(2), 16.1% Al(2)O(3), 10.9% K(2)O, 4.3% Na(2)O, 1.7% CaO, 0.5% LiO(2) and 0.4% TiO(2) was heated in an electric furnace and later quenched to produce glasses. The glass powders were ball milled to two different particle sizes and heat-treated using one and two-step crystallization heat treatments. Dta, Xrd, and Sem analyses was used to characterise and explore the crystallization kinetics of the glasses. RESULTS: Selected heat treatments of the glass powders produced a uniform distribution of fine tetragonal leucite crystals (mean+/-S.D.) 0.1+/-0.2 microm(2) in the glassy matrix, with minimal matrix microcraking in the glass ceramics produced. The addition of a two-step heat treatment increased the leucite volume fraction in all instances. SIGNIFICANCE: Selected crystallization heat treatments and powder particle sizes were used to control the leucite crystal size, distribution and volume fraction, in order to produce uniformly distributed ultra fine grained tetragonal leucite glass-ceramics for dental applications. PMID- 16375964 TI - Effect of whitening gels on the surface roughness of restorative materials in situ. AB - Peroxide gels are effective in changing tooth colour but their effect on restorative materials has been poorly studied. The purpose of this investigation was to assess the impact of a commercially available whitening gel containing hydrogen peroxide and a sodium percarbonate formulation on the surface of restorative materials. A total of 12 subjects participated in a double-blinded crossover study. Each wore an intra-oral appliance containing five bovine enamel blocks restored with amalgam, posterior composite, microfilled composite, glass ionomer cement and porcelain. Appliances were worn continuously for 14 days and whitening products were applied twice daily. After 14 days the appliances were removed and values for roughness (R(a)) were obtained using atomic force microscopy. Mean values of R(a) were assessed between baseline and 14 days, and although minor variations were seen, there were no statistically significant differences detected for any material or any whitening product. PMID- 16375965 TI - Photolithographic patterning of polyethylene glycol hydrogels. AB - A simple, inexpensive photolithographic method for surface patterning deformable, solvated substrates is demonstrated using photoactive poly(ethylene glycol) (PEG) diacrylate hydrogels as model substrates. Photolithographic masks were prepared by printing the desired patterns onto transparencies using a laser jet printer. Precursor solutions containing monoacryloyl-PEG-peptide and photoinitiator were layered onto hydrogel surfaces. The acrylated moieties in the precursor solution were then conjugated in monolayers to specific hydrogel regions by exposure to UV light through the transparency mask. The effects of UV irradiation time and precursor solution concentration on the levels of immobilized peptide were characterized, demonstrating that bound peptide concentration can be controlled by tuning these parameters. Multiple peptides can be immobilized to a single hydrogel surface in distinct patterns by sequential application of this technique, opening up its potential use in co-cultures. In addition, 3D structures can be generated by incorporating PEG-diacrylate into the precursor solution. To evaluate the feasibility of using these patterned surfaces for guiding cell behavior, human dermal fibroblast adhesion on hydrogel surfaces patterned with acryloyl-PEG-RGDS was investigated. This patterning method may find use in tissue engineering, the elucidation of fundamental structure-function relationships, and the formation of immobilized cell and protein arrays for biotechnology. PMID- 16375966 TI - Adhesion contact dynamics of 3T3 fibroblasts on poly (lactide-co-glycolide acid) surface modified by photochemical immobilization of biomacromolecules. AB - A simple and effective method of biomacromolecule immobilization on biomaterial surface for direct tuning of biophysical parameters such as the initial cell deformation rate, degree of cell spreading and adhesion kinetics is important for tissue engineering. The photochemical immobilization of azide-chitosan (Az-CS) on poly (lactide-co-glycolide) acid (PLGA) is applied here. Chitosan immobilization on PLGA through the photoactive azide group further facilitates subsequent grafting of other biocompatible biomacromolecules like gelatin (Gel) through the active amine groups on CS. This study quantitatively compares the 3T3 fibroblast adhesion dynamics on three PLGA surfaces (Gel-CS-PLGA, CS-PLGA and unmodified PLGA surfaces) using Confocal-Reflectance Interference Contrast Microscopy (C RICM) together with phase contrast imaging. CS-PLGA and Gel-CS-PLGA surfaces developed were confirmed by X-ray photoelectron spectroscopy, atomic force microscopy and water contact angle and cell adhesion contact dynamics measurements. The cell adhesion was strongest on the Gel-CS-PLGA surface and lowest on unmodified PLGA. The steady state adhesion energy attained by the cells on gelatin modified PLGA surface is determined as 4.0 x 10(-8) J/m(2), which is about 400 times higher than that on PLGA surface (1.1 x 10(-10) J/m(2)). Significantly increased cell adhesion with Gel-CS-PLGA is postulated to result in increased cell spreading. Our integrated biophysical method can quantify the transient contact dynamics and is sufficiently accurate to discriminate even between Gel and CS modified surfaces. PMID- 16375967 TI - Expression of cyclooxygenase-II (COX-II) and 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD)/prostaglandin F-synthase (PGFS) in bovine placentomes: implications for the initiation of parturition in cattle. AB - The chain of events leading to prepartal luteolysis in cattle is not yet fully understood. Prostaglandin F(2alpha) (PGF(2alpha)) seems to be a major factor involved. However, only little information is available about the underlying regulatory mechanisms. Consequently, the expression of cyclooxygenase-II (COX-II) and 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD), an enzyme recently shown to be most likely responsible for the production of luteolytic PGF(2alpha) in the endometrium of cyclic cows, was investigated in bovine placentomes. Immunohistochemical methods were applied to placentomes from 17 pregnant cows between days 100 and 284, from three cows during the prepartal progesterone decrease (days 273-282) and from five parturient cows. COX-II was found in uninucleated trophoblast cells (UTC) from day 100 until parturition. However, between days 100 and 235 expression was only weak to moderate, focal and mainly restricted to the chorionic plate and adjacent basal parts of chorionic stem villi. In placentomes from a 270 and a 284 day pregnant cow, in which the prepartal decline of progesterone levels had not started yet, staining had substantially increased and extended to secondary and tertiary chorionic villi. In prepartal and parturient cows strong to intense staining was present in UTC all over the villous tree. Real time RT-PCR confirmed an extensive pre- and intrapartal rise of COX-II expression in bovine placentomes with a 70-100-fold increase of COX-II-mRNA levels. 20alpha-HSD/PGFS was widely expressed in UTC of chorionic villi. Like COX-II it was down-regulated in UTC differentiating into trophoblast giant cells. Immunostaining pattern did not change remarkably during the period under investigation, and 20alpha-HSD/PGFS-mRNA levels increased only 2.6-fold in the prepartal phase. Thus, in UTC PGF(2alpha) may be produced via COX II and 20alpha-HSD/PGFS, but only COX-II may be substantially involved in the control of a putative prepartal placentomal output of luteolytic PGs, whereas 20alpha-HSD/PGFS seems to be expressed in a more constitutive manner. PMID- 16375968 TI - Inhibition of water intake by the central administration of IL-1beta in rats: role of the central opioid system. AB - In the present study we investigated, the effect of third ventricle injections of IL-1beta on water intake, in rats, induced by three different physiological stimuli: dehydration induced by water deprivation, hypernatremia associated with hyperosmolarity induced by intragastric salt load, and hypovolemia produced by subcutaneous polytethyleneglycol administration. Central administration of IL 1beta at the doses of 4 and 8 ng reduced water intake in all three conditions studied. Third ventricle injections of IL-1beta (8 ng) were unable to diminish water intake in the groups of rats pretreated with central injections of the non selective opioid antagonist naloxone (10 microg) in the three different conditions studied. Furthermore, the central administration of IL-1beta was neither able to modify the intake of a 0.1% saccharin solution when the animals were submitted to a "dessert test" nor to induce any significant locomotor deficit in the open-field test. These results suggest that the central activation of interleukin-1 receptors by IL-1beta is able to impair the thirst-inducing mechanisms triggered by the physiological stimuli represented by dehydration, hyperosmolarity and hypovolemia. These results lead us to conclude that the antidipsogenic effects observed following central administration of IL-1beta require the functional integrity of the brain opiatergic system. PMID- 16375969 TI - CD5+ B cells are preferentially expanded in rabbit appendix: the role of CD5 in B cell development and selection. AB - Although only a small proportion of mouse and human B cells are CD5(+), most adult rabbit B cells express CD5. However, CD5 was not detectable on the majority of B cells in neonatal appendix 1 and 3days after birth. Cell trafficking studies demonstrated that CD5(+) and CD5(-) CD62L(+) B cells from bone marrow migrated into appendix. There, CD5(+) B cells were preferentially expanded and predominated by approximately 2weeks of age. In mutant ali/ali rabbits, VHa2(+) B cells develop through gene conversion-like alteration of rearranged VH genes upstream of deleted VH1a2. Correlated appearance of individual CD5(+) germinal centers and VHa2(+) B-cells in mutant appendix suggests that CD5 binding positively selects cells with a2(+) framework regions that bind CD5. Following negative and positive selection, cells with diversified rearranged heavy- and light-chain sequences exit appendix, migrate to peripheral tissues and constitute the preimmune repertoire of CD5(+) B cells that encounter foreign antigens. PMID- 16375970 TI - Biological and molecular docking studies on coagulin-H: Human IL-2 novel natural inhibitor. AB - Withanolide, coagulin-H (1), was evaluated for its effect on various cellular functions related to immune response including lymphocyte proliferation, and expression of interleukin-2 (IL-2) cytokine, and results were compared with prednisolone (2), a commonly used immune modulating drug. Coagulin-H (1) was found to have a powerful inhibitory effect on lymphocyte proliferation and Th-1 cytokine production. Inhibition of the phytohaemagglutinin (PHA)-activated T-cell proliferation by coagulin-H (1) was observed in a concentration dependent manner. A complete suppression of PHA-activated T-cell was observed at > or =2.5 microg/mL concentrations of compound (1) and this suppression activity was similar to that of prednisolone (2). Coagulin-H (1) also significantly inhibited IL-2 production by 80%. The interactions of coagulin-H (1) (a natural inhibitor) and prednisolone (2) (a drug) to IL-2 were also investigated in order to understand the differences in their effects on T-cell responses. This paper also describes the results of molecular docking study on IL-2 inhibition. Docking studies predicted that coagulin-H (1) binds to receptor binding site of IL-2 more effectively than prednisolone (2). Based on the computational and the experimental results, coagulin-H (1) was identified as a potential immunosuppressive candidate. PMID- 16375971 TI - Burkholderia respiratory tract infections in Italian patients with cystic fibrosis: molecular characterization. AB - OBJECTIVES: To characterize by molecular techniques Burkholderia strains responsible for respiratory tract infections in cystic fibrosis (CF) patients (children and adults), to assign the Burkholderia cepacia complex (Bcc) isolates to a genomovar and to assess the presence of cblA and esmR genes in bacteria. Unique or sequential Burkholderia isolates (n=48) that had been collected from eight CF children and 17 adults over several (4-6) years were investigated; moreover 11 reference strains were analyzed. METHODS: The microorganisms were identified by using biochemical methods, genotyped by pulse field gel electrophoresis (PFGE) and random-amplified polymorphic DNA fingerprinting-PCR (RAPD-PCR), and assessed by PCR assays for the genomovar and cblA and esmR genes of Bcc. RESULTS: Among isolates 70.8% were identified as Bcc genomovar III-A; one child was infected by Burkholderia ambifaria and four adults were colonized with Burkholderia gladioli. The cblA gene was not detected in any of the isolates, while the esmR gene was detected in the 52.1% of the strains, all belonging to genomovar III-A. CONCLUSION: Molecular analysis of strains revealed in CF patients a colonization with a persistent Burkholderia flora with strains of one genotype. The prevalence of Bcc of genomovar III-A in the two categories of patients and of B. gladioli in four adults demonstrated that transmission may have occurred between subjects. Moreover the B. ambifaria infection demonstrated in a child may be environmentally derived. PMID- 16375972 TI - The influence of age and gender on the population-based incidence of community acquired pneumonia caused by different microbial pathogens. AB - OBJECTIVES: To determine the population-based incidence of community-acquired pneumonia (CAP) in adults and to assess the relative importance of age and gender on the incidence of infections caused by different microbial pathogens. METHODS: A two-year prospective study in a well-defined geographic area of the Spanish Mediterranean coast. RESULTS: The overall incidence rate of CAP was 12 cases (95% CI 11.25-13.45) per 10,000 person-years. Incidence rates increased by age (p<0.0001) and they were higher in males (16 versus 9 cases per 10,000 person years; p<0.0001). The rate was especially high among males aged > or = 75 years (87 cases per 10,000 person-years). The incidence of pneumococcal pneumonia increased significantly with ageing and it was particularly high among people aged > or = 75 years (10 cases per 10,000 person-years). Very elderly people had also a 15-fold higher incidence of CAP associated with influenza virus and a 5 fold higher incidence of infections by Chlamydophila spp., than young adults. The incidence of infections with Legionella pneumophila also increased with age and it was 10 times higher in males. In contrast, the incidence of pneumonia caused by Mycoplasma pneumoniae was unrelated to age and gender. CONCLUSIONS: Age and gender have a strong influence on the overall incidence of CAP and on the incidence of pneumonia caused by the main microbial pathogens, including not only Streptococcus pneumoniae, but also influenza virus, Chlamydophila spp. and L. pneumophila. Ageing is associated with a higher risk of acquiring pneumonia by S. pneumoniae, influenza virus and Chlamydophila spp., whereas male gender increases greatly the incidence of L. pneumophila and Chlamydophila spp. PMID- 16375973 TI - Emergence of cefotaxime resistance in Citrobacter freundii causing necrotizing fasciitis and osteomyelitis. AB - We present a diabetic patient with necrotizing fasciitis caused by Citrobacter freundii associated with an injury from the fish fin. Two isolates recovered six days after cefotaxime treatment had a cefotaxime minimum inhibitory concentration (MIC) of 0.12 and 256 mg/L, respectively, and a cefepime MIC of 0.03 and 0.25 mg/L, respectively. The two isolates both possessed bla(CMY-2) gene. The patient responded unsatisfactorily to cefotaxime and cefepime therapy and surgical debridement but recovered completely after ertapenem treatment for 42 days. PMID- 16375974 TI - Smaller volume of anterior cingulate cortex in abuse-related posttraumatic stress disorder. AB - OBJECTIVE: The anterior cingulate cortex (ACC) plays an important role in emotion, and studies in animals have shown changes in ACC structure with early life stress. The purpose of this study was to measure volume of the ACC in PTSD. METHOD: Magnetic resonance imaging (MRI) was used to measure ACC volume in 8 subjects with abuse-related PTSD and 13 healthy subjects without PTSD. ACC volume included Brodmann's area [BA] 24 and 32. RESULTS: Right ACC volume in PTSD patients was significantly smaller than in non-PTSD subjects. CONCLUSION: These results are consistent with smaller ACC volume in PTSD. PMID- 16375975 TI - Breaking or making immunological privilege in the central nervous system: the regulation of immunity by neuropeptides. AB - Immune privilege in the central nervous system (CNS) is not maintained by immune ignorance of the CNS, but by CNS control over inflammatory processes. In this review we examine the role neuropeptides play in maintenance of immune privilege in the CNS. Vasoactive intestinal peptide, alpha-melanocyte-stimulating-hormone, neuropeptide Y, and somatostatin are members of an anti-inflammatory repertoire of immune modulators, while substance P acts to break immune privilege and promote inflammation in the CNS. Here we focus both on cellular responses to these neuropeptides and the role these peptides play in immune privilege as it relates to CNS autoimmunity. PMID- 16375977 TI - Neural-immune interactions: an integrative view of the bidirectional relationship between the brain and immune systems. AB - This review briefly summarizes a part of the relevant knowledge base of neuroimmunology, with particular emphasis on bidirectional neural-immune interactions. These complex systems interact at multiple levels. Both neuroendocrine (the primary hormonal pathway is hypothalamic-pituitary-adrenal axis) and neuronal (direct sympathetic innervation of the lymphoid organs) pathways are involved in the control of the humoral and cellular immune responses. Although, the recent evidence has been made on immunosuppressive effect of acetylcholine-secreting neurons of the parasympathetic nervous system. The immune system, in turn, influences the central nervous system primarily through cytokines. At the molecular level, neuro- and immune signal molecules (hormones, neurotransmitters, neuropeptides, cytokines) or their receptors are member of the same superfamily which enable the mutual neuroimmune communication. Most extensively studied are cytokine-neuropeptide/neurotransmitter interactions and the subcellular and molecular mechanisms of these interactions. At the system (neuroanatomical) level, advances in neural-immune communication have been made in the role of discrete brain areas related to emotionality. The immunoenhancement, including the antiviral and antitumor cytotoxic activity, related to the "brain reward system", limbic structures and neocortex, offers a new directions for therapy in immune disorders. PMID- 16375976 TI - Raft localisation of FcgammaRIIa and efficient signaling are dependent on palmitoylation of cysteine 208. AB - Ligand-dependent aggregation of FcgammaRIIa initiates multiple biochemical processes including the translocation to detergent resistant membrane domains (DRMs) and receptor tyrosine phosphorylation. Palmitoylation of cysteine residues is considered to be one process that assists in the localisation of proteins to DRMs. Within the juxtamembrane region of FcgammaRIIa there is cysteine residue (C208) that we show to be palmitoylated. Mutation of this cysteine residue results in the disruption of FcgammaRIIa translocation to DRMs as empirically defined by insolubility at high Triton X-100 concentrations. This study also demonstrates that the lack of lipid raft association diminishes FcgammaRIIa signaling as measured by receptor phosphorylation and calcium mobilisation functions suggesting that FcgammaRIIa signaling is partially dependent on lipid rafts. PMID- 16375978 TI - IFN-gamma production and astrocyte recognition by autoreactive T cells induced by Theiler's virus infection: role of viral strains and capsid proteins. AB - From mice infected with the DA strain of Theiler's murine encephalomyelitis virus (TMEV), CD8+ cytotoxic T lymphocytes (CTLs) could be detected after stimulation with TMEV infected antigen presenting cells (APCs). These CTLs killed not only TMEV infected but also uninfected syngeneic cells. Killing was associated with interferon (IFN)-gamma production in ELISPOT assays. The CTLs were efficiently induced by vaccinia virus encoding DA virus capsid proteins, but not by APCs infected with the GDVII strain of TMEV. The CTLs produced IFN-gamma in response to TMEV infected, but not uninfected, astrocytes. The CTLs could be maintained in the presence of interleukin (IL)-2. We hypothesized that, in DA virus infection, CD8+ CTLs specific for viral capsid protein could recognize self protein on oligodendrocytes by molecular mimicry, leading to demyelination. PMID- 16375979 TI - Liposome-mediated transient transfection reduces cholesterol-dependent coxsackievirus infectivity. AB - Liposome-mediated gene delivery provides a powerful strategy for the study of gene function and for gene therapy. Coxsackievirus B3 is an important human pathogen associated with various diseases. Here we reported that liposome mediated transient transfection of plasmid cDNA inhibited coxsackieviral replication at the levels of RNA, protein and viral progeny release. These inhibitory effects were observed in various cell types and by using different liposome reagents. We further showed that the inhibition was likely due to the lack of virus attachment. Moreover, we showed that addition of cholesterol restored, at least in part, the viral infectivity. Interestingly, we found that membrane cholesterol levels were unchanged during transfection, indicating that disruption rather than depletion of membrane cholesterol contributes to the inhibitory effects of transfection. Our data suggest that liposome-mediated cDNA transient transfection inhibits coxsackievirus infectivity via inhibition of viral attachment, which is likely occurring through the changes of membrane cholesterol integrity. PMID- 16375980 TI - A sensitive in-house RT-PCR genotyping system for combined detection of plasma HIV-1 and assessment of drug resistance. AB - Quantification of the viral burden and identification of drug resistant mutations are important laboratory tools in the management of HIV-1 infected patients. However, widespread use of assays for viral load determination and genotyping is still hampered by the high cost. Here, an in-house RT-PCR-sequencing assay for HIV-1 drug resistance monitoring with the potential to be used both as a qualitative assay to detect the virus in plasma and as a genotyping system is described. A total of 377 clinical samples, collected from 374 HIV-infected patients of diverse geographic origin, were tested. The nested RT-PCR for amplification of the protease reverse transcriptase gene was found positive for 350 (92.8%) and 346 (91.8%) of 377 samples, respectively. All amplification failures were due to viral loads of below 500 copies/ml. However, low viral load does not exclude amplification since 80.2 and 76% of 121 samples with viral loads of less than 500 copies/ml were amplified successfully for protease and reverse transcriptase, respectively. The high sensitivity of the assay was independent of the HIV-subtype, with a broad range of different HIV-1 subtypes tested. In conclusion the RT-PCR-direct sequencing method is convenient for the sensitive detection and subsequent genotyping of plasma RNA from a broad range of different HIV-1 subtypes. The assay enables the accurate follow-up of patients under treatment at a significantly reduced cost compared to the currently available commercial assays for viral load assessment and genotyping. PMID- 16375981 TI - Fenretinide inhibits HIV infection by promoting viral endocytosis. AB - HIV fusion is mediated by the sequential interaction of the viral envelope glycoprotein with cellular receptors at the plasma membrane. We have previously reported that the upregulation of cellular ceramide levels following fenretinide treatment inhibits HIV fusion. As ceramide facilitates the internalization of a variety of microbes, we hypothesized that it may also promote the engulfment of HIV virions. Hence, we analyzed the effect of fenretinide treatment on virus binding and uptake. We observed that virus binding is not altered by fenretinide treatment. The distribution of HIV receptors was also unchanged. In contrast, virus uptake showed a significant increase. We have determined that fenretinide treatment promotes the internalization of virions from the plasma membrane and the accumulation of virus in the endocytic fraction of HeLa cells. This effect of fenretinide appears to be specific for virus as the endosomal accumulation of gp120, transferrin and horse-radish peroxidase was not increased. Notably, fenretinide increased the infectivity of influenza virus, which fuses in the endosomal compartment upon low pH activation. Our data suggest that fenretinide treatment effectively inhibits HIV infection by re-directing the virus to the endocytic pathway. PMID- 16375982 TI - Disruption of the bradyzoite-specific P-type (H+)-ATPase PMA1 in Toxoplasma gondii leads to decreased bradyzoite differentiation after stress stimuli but does not interfere with mature tissue cyst formation. PMID- 16375983 TI - Somatostatin inhibits dendritic cell responsiveness to Helicobacter pylori. AB - Somatostatin is a regulatory peptide found in abundance in the stomach. We have previously shown that somatostatin is required for IL-4-mediated resolution of Helicobacter pylori gastritis. In the current study, we hypothesize that somatostatin acts directly on antigen-presenting cells in the stomach to lessen the severity of gastritis. To test this hypothesis, we first show that CD11c+ dendritic cells are present in the infected tissue of mice with H. pylori-induced gastritis. Pretreatment of bone marrow-derived dendritic cells with somatostatin results in decreased IL-12 production, and lower splenocyte proliferation induced by H. pylori-stimulated dendritic cells. Furthermore, octreotide, a somatostatin analogue, is more potent than somatostatin in suppressing IL-12 release by H. pylori-stimulated dendritic cells through an NF-kappaB-independent pathway. In addition, IL-4 stimulates somatostatin secretion from dendritic cells. In conclusion, somatostatin inhibits dendritic cell activation by H. pylori; a possible mechanism by which IL-4 mediates resolution of gastritis. We suggest that octreotide may be effective in treating immune-mediated diseases of the stomach. PMID- 16375984 TI - QT dispersion in comparison to Tl-201-SPECT for detection of myocardial ischaemia. AB - OBJECTIVE: There is controversy whether the measurement of QT dispersion might provide information about the presence and extent of ischaemic myocardium. It was the aim to analyse the diagnostic value of QT dispersion for the detection of myocardial ischaemia in comparison to Tl-201-SPECT. PATIENTS AND METHODS: 100 patients were randomly selected from patients referred for Tl-201-SPECT. QT duration was measured in all 12 leads in the resting ECG and in the ECG either at maximal bicycle exercise or during peak dipyridamole stress. QT dispersion was calculated and correlated with parameters of ischaemia for all patients and for the bicycle exercise and the dipyridamole group separately. RESULTS: 25 patients had to be excluded from the analysis because QT dispersion could not be measured. Regression analysis did not show significant correlation neither for all 75 patients nor for the bicycle exercise nor for the dipyridamole group correlating grade of myocardial ischaemia, number of ischaemic segments and summed ischaemic stress score with QT dispersion in the resting or the stress ECG and QTd and there was no significant difference between groups. CONCLUSION: QT dispersion could not be measured in 25/100 patients (25%). In the remaining patients QT dispersion did not correlate with the extent or grade of myocardial ischaemia. PMID- 16375985 TI - The endophytic fungus Trametes hirsuta as a novel alternative source of podophyllotoxin and related aryl tetralin lignans. AB - The aryl tetralin lignans are synthesized by Podophyllum sps. and are in great demand worldwide due to their use in synthesis of topoisomerase inhibitors. However, the sustained production of these aryl tetralin lignans requires large scale harvesting from the natural environments, which has resulted in the plant endangered status. In view of the difficulties in their total chemical synthesis, cultivation and failure of metabolic engineering approaches, there is a need to search for alternative sources of production of aryl tetralin lignans. We unequivocally established the methodology for isolation, identification, and characterization of a novel fungal endophyte (Trametes hirsuta) that produces aryl tetralin lignans consistently as shown by HPLC, LC-MS, LC/MS-MS and (1)H NMR. The lignans produced by the microorganism are biologically active, and exhibit potent antioxidant, anticancer and radioprotective properties. This strategy promises to improve the production of these therapeutically important compounds at lower costs. PMID- 16375986 TI - Well-ordered mesoporous bioactive glasses (MBG): a promising bioactive drug delivery system. AB - The local drug release system is considered to be an alternative to treat the bone infection. In this paper, well-ordered mesoporous bioactive glasses (MBG) with high specific surface area have been synthesized in aqueous solution by a two-step acid-catalyzed self-assembly process combined with hydrothermal treatment. Gentamicin was encapsulated into the MBG by adsorption method and in vitro release of gentamicin from MBG was performed in distilled water and modified simulated body fluid (SBF), respectively. The results showed that the amount of drug loading of MBG was three times more than that of conventional sol gel 58S. The outcomes of drug release in distilled water and in SBF showed that M58S effectively decreased the initial burst. During the release period, gentamicin was released from the M58S at a much lower release rate as compared to that from 58S after soaking in distilled water and SBF. Furthermore, the drug release was sensitive to the pH and ionic concentration of the release medium suggesting possible controls of the release rate. In addition, in contrast to conventional sol-gel 58S, M58S had higher ability to induce hydroxyapatite (HAp) formation. Therefore, well-ordered mesoporous bioactive glasses might be used as a bioactive drug release system for preparation of bone implant materials. PMID- 16375987 TI - Prognostic value of DNA ploidy: 5-year follow-up of patients with resectable squamous cell carcinoma (SCC) of the lung. AB - This study was designed to show the relation between DNA-ploidy in patients with resectable lung cancers and their 5-year survival rate. The results are compared with those from our 2-year follow-up study of the same group of patients published in 2000. The group of 80 patients with SCC who underwent lung resection between 1995 and 96 were re-analyzed. For the statistical analysis the hazard Cox model and an exponential multiple regression model were used. The survival curves were drawn using the Kaplan-Meier method. DNA-aneuploidy was found in 45% of cancer tumors. There was no statistically significant correlation between aneuploidy and gender, age, cancer staging or grading. In the 3-year follow-up the survival rate in patients with aneuploid type tumors was significantly lower than in those with the diploid type. However, this difference was not found after 5 years of follow-up. Tumor ploidy was an independent prognostic factor only in patients between 55 and 60 years of age. The mortality rate in patients with aneuploid tumors was mainly the result of distant metastases while, in patients with diploid tumors, local recurrence was the main reason for death. In the first three years after surgical resection patients with aneuploid tumors are at higher risk of distant metastases than patients with the diploid type. Tumor ploidy can be recognized as an independent prognostic factor in younger (55-60) patients. Aneuploidy promoted the occurrence of early distant metastases while the diploid type was associated with late (after 3 years) local tumor recurrence. PMID- 16375989 TI - Pharmacology of the human CGRP1 receptor in Cos 7 cells. AB - Only limited pharmacological characterization of the CGRP1 receptor, a heterodimer of the calcitonin (CT) receptor-like receptor (CL) and receptor activity-modifying protein 1 has been performed in cells that do not endogenously express RAMP2. We characterized the receptor in RAMP-deficient Cos 7 cells by measuring cAMP responses following agonist treatment in the absence or presence of antagonists. Potent cAMP responses to human alpha-and beta-CGRP (Cys(Et)2,7)halphaCGRP and human adrenomedullin (AM) were observed. Adrenomedullin15-52 was also an effective agonist of the CGRP1 receptor but human and salmon calcitonin and rat amylin were only weak agonists. As expected, BIBN4096BS and CGRP(8-37) were effective antagonists of the CGRP1 receptor. (Cys(Acm)2,7)halphaCGRP also antagonized CGRP responses. Antagonists of related receptors were only weakly able to inhibit CGRP responses. PMID- 16375990 TI - The effect of heme oxygenase-1 induction by octreotide on radiation enteritis. AB - Radiation enteritis occurs as a response to abdominal radiation, which can cause mucosal damage in the gastrointestinal mucosal epithelium. The small intestine is one of the most radiosensitive organs in the abdomen. The present study was undertaken to investigate the effect of octreotide (OCT) administration on heme oxygenase-1 (HO-1) expression of the radiation enteritis model. Rats received 50 mg/kg/day OCT for 4 days before irradiation and continued for 3 days after irradiation. Intestinal myeloperoxidase (MPO) activities, malondialdehyde (MDA) levels are indicators of oxidative damage while caspase-3 activities reveal apoptosis degree of the small intestine. At histological examination, the terminal ileum tissue was analyzed for morphological changes. Irradiation significantly increased the intestinal MPO and caspase-3 activities, MDA levels and HO-1 expression in comparison to sham control group. OCT treatment was associated with increased HO-1 expression and caspase-3 activity, decreased MPO activity and MDA levels. Histological examination revealed that the intestinal mucosal structure was preserved in the OCT treated group. OCT appears to have protective effects against radiation-induced intestinal damage. This protective effect is, in part, mediated by modification of the inflammatory response and the induction of HO-1 expression. PMID- 16375991 TI - 3-Bromo-4-(1H-3-indolyl)-2,5-dihydro-1H-2,5-pyrroledione derivatives as new lead compounds for antibacterially active substances. AB - A number of new compounds containing 3-bromo-2,5-dihydro-1H-2,5-pyrroledione and indole substructures were found to have antibacterial activity against resistant strains of Staphylococcus aureus, Mycobacterium smegmatis and some other Gram positive bacteria. The investigated compounds exhibit minimal inhibition concentrations (MIC's) lower than those of ciprofloxacin, vancomycin and doxycycline resp. A different spectrum of activity, suggests a mechanism of action different to vancomycin and doxycycline. This might be important in circumventing existing resistance mechanisms. Here we report about the synthesis and on the antibacterial activity in a structure activity relationship study. PMID- 16375992 TI - Nonsteroidal antiinflammatory agents-part 2 antiinflammatory, analgesic and antipyretic activity of some substituted 3-pyrazolin-5-ones and 1,2,4,5,6,7-3H hexahydroindazol-3-ones. AB - As a part of a research project on the synthesis of a number of substituted 1 (pyrimidin-2-yl)-3-pyrazolin-5-ones and 2-(pyrimidin-2-yl)hexahydroindazol-3-ones and as a result of the interesting antiinflammatory, analgesic and antipyretic activities recorded for some of these compounds, some new 3-pyrazolin-5-ones and hexahydroindazol-3-ones linked to substituted imidazolyl, pyrimidyl and tetrahydroquinazolinyl moieties were prepared and evaluated for such activity (). A structure-activity relationship (SAR) comparative study indicated that some compounds from 3-pyrazolin-5-one (2, 6-8, 10) and indazolone (18, 20, 24, 27, 29) series exhibited pronounced antiinflammatory, analgesic and antipyretic activities relative to indomethacin. Most of these compounds were found to be nearly equipotent in the antiinflammatory screen (ED(50)=16.8-19.9 mg/kg) whereas the lead compound, 2-indazolyl-4-pyrimidineacetic acid 24 (), was found to be the most potent among this series (ED(50)=9.9 mg/kg). Additionally, the most active compounds were shown to have a large safety margin (ALD(50)=3.0 g/kg, po) and devoid of ulcerogenic potentialities when administered orally at a dose of 300 mg/kg. PMID- 16375993 TI - Oligosaccharides isolated from goat milk reduce intestinal inflammation in a rat model of dextran sodium sulfate-induced colitis. AB - BACKGROUND AND AIMS: There is increased interest in the study of manipulation of the flora with pro- and prebiotics regarding inflammatory bowel disease. The aim of this work was to evaluate the effect of oligosaccharides from goat milk in a rat model of dextran sodium sulfate- (DSS-) induced colitis. METHODS: Twenty rats were fed the same diet but with different sources of fiber (5% of the diet): cellulose or a mixture of goat's milk oligosaccharides (GMO) and cellulose. DSS treatment was used to induce a colonic inflammation. Several clinical and inflammatory parameters, as well as intestinal micorbiota and gene expression by DNA microarray technology, were evaluated. RESULTS: DSS induced a decrease in body weight which was not observed in rats fed the GMO (decrease of 21+/-11% in control rats vs increase of 5.2+/-8.6 in GMO rats, P<0.05). DSS also caused an acute colonic inflammatory process which was weaker in rats fed the GMO, as shown by colon myeloperoxidase activity (0.53+/-0.16 vs 0.14+/-0.07U/mg of protein, P<0.05), as well as clinical symptoms measured by a scoring system (1.25+/-1.14 vs 0.4+/-0.07, P<0.05). GMO rats also showed less severe colonic lesions and a more favorable intestinal microbiota. The expression of genes involved in intestinal function, such as mucine-3, was down-regulated in DSS-control rats but returned to normal values in GMO rats. CONCLUSION: GMO reduce intestinal inflammation and contribute to the recovery of damaged colonic mucosa. PMID- 16375994 TI - Comparison of body weight and composition measured by two different dual energy X ray absorptiometry devices and three acquisition modes in obese women. AB - BACKGROUND AND AIMS: Weight measured by dual-energy X-ray (DXA) was shown to be increasingly underestimated in subjects over 75 kg compared to an electronic scale. This study compares body weight and composition measured by balance beam scale and three DXA acquisition modes in obese subjects. METHODS: In 39 obese, body weight was measured by balance beam scale, and body weight and composition by DXA Hologic QDR4500A in normal (NPM) and high power mode (HPM) (Enhanced v8.26 and v8.26* software) and DXA GE-Lunar Prodigy (v6.5 software). To ensure linearity of body weight and composition measured by the different DXA acquisitions, we also measured 13 women with a body mass index (BMI) of 25-30 kg/m(2). RESULTS: While QDR4500A HPM overestimates scale weight by about 2 kg over the whole BMI spectrum, QDR4500A NPM underestimates scale weight as a weight dependent response (-1.7+/-1.8 kg overall, -4.1+/-1.6 kg in morbidly obese women). These results suggest switching from one mode to the other at a specific threshold, i.e. in our study a weight of 90 kg or a BMI of 34 kg/m(2). Prodigy gives weight about similar to scale (+0.5+/-0.8 kg). Both Hologic acquisition modes underestimate fat mass but overestimate lean body mass compared to Prodigy. CONCLUSIONS: The QDR4500A NPM is inappropriate in women over 90 kg. Unfortunately, the QDR4500A HPM overestimates body weight in the range of 90-150 kg. The difference between scale and Prodigy weight remains stable throughout weight ranges. To better assess their accuracies in terms of body composition, QDR4500A NPM, HPM and Prodigy should be tested against phantoms or in vivo multi compartment models. PMID- 16375995 TI - Trivalent inactivated influenza vaccine safety in children: assessing the contribution of telephone encounters. AB - We assessed the contribution of telephone medical care encounters to surveillance of adverse events (AE) following trivalent influenza vaccination in children age 6 months to 17 years. We used retrospective, self-controlled, case-series analysis to estimate adverse event incidence rate ratios for post-vaccination risk intervals relative to 15-28 days prior to vaccination. We confirmed possible vaccination reactions by medical record abstraction. Detection of 10 of 20 elevated incidence rate ratios required telephone data. We conclude that telephone encounters substantially contribute to the detection of possible influenza vaccination reactions, primarily local injection site and systemic reactions. PMID- 16375996 TI - Conceptual approaches for the development of dynamic specific activity models of 14C transfer from surface water to humans. AB - Carbon-14 is a particularly interesting radionuclide from the perspective of dose estimation. Many nuclear facilities, including power reactors, release 14C into the environment, and much of this is as 14CO2. This mixes readily with stable CO2, and hence enters the food chain as fundamental biomolecules. This isotopic mixing is often used as the basis for dose assessment models. The present model was developed for the situation of 14C releases to surface waters, where there are distinct changes in the water 14C activity concentrations throughout the year. Complete isotopic mixing (equilibrium) cannot be assumed. The model computes the specific activity (activity of 14C per mass of total C) in water, phytoplankton, fish, crops, meat, milk and air, following a typical irrigation based food-chain scenario. For most of the biotic compartments, the specific activity is a function of the specific activity in the previous time step, the specific activity of the substrate media, and the C turnover rate in the tissue. The turnover rate is taken to include biochemical turnover, growth dilution and mortality, recognizing that it is turnover of C in the population, not a tissue or an individual, that is relevant. Attention is paid to the incorporation of 14C into the surface water biota and the loss of any remaining 14CO2 from the surface water-air interface under its own activity concentration gradient. For certain pathways, variants in the conceptual model are presented, in order to fully discuss the possibilities. As an example, a new model of the soil-to-plant specific activity relationship is proposed, where the degassing of both 14C and stable C from the soil is considered. Selection of parameter values to represent the turnover rates as modeled is important, and is dealt with in a companion paper. PMID- 16375997 TI - Partitioning of radionuclides and trace elements in phosphogypsum and its source materials based on sequential extraction methods. AB - Phosphogypsum is a waste produced by the phosphate fertilizer industry. Although phosphogypsum is mainly calcium sulphate dihydrate, it contains elevated levels of impurities, which originate from the source phosphate rock used in the phosphoric acid production. Among these impurities, radionuclides from 238U and 232Th decay series are of most concern due to their radiotoxicity. Other elements, such as rare earth elements (REE) and Ba are also enriched in the phosphogypsum. The bioavailability of radionuclides (226Ra, 210Pb and 232Th), rare earth elements and Ba to the surrounding aquatic system was evaluated by the application of sequential leaching of the phosphogypsum samples from the Brazilian phosphoric acid producers. The sequential extraction results show that most of the radium and lead are located in the "iron oxide" (non-CaSO4) fraction, and that only 13-18% of these radionuclides are distributed in the most labile fraction. Th, REE and Ba were found predominantly in the residual phase, which corresponds to a small fraction of the phosphate rock or monazite that did not react and to insoluble compounds such as sulphates, phosphates and silicates. It can be concluded that although all these elements are enriched in the phosphogypsum samples they are not associated with CaSO4 itself and therefore do not represent a threat to the surrounding aquatic environment. PMID- 16375998 TI - Interesting case: an unusual injury to the scalp by a sea urchin. PMID- 16375999 TI - Trismus: an unconventional approach to treatment. AB - A 63 year-old man who had had an operation for adenocarcinoma of the left parotid gland presented with worsening trismus after the completion of radiotherapy. None of the treatments given were successful so he resorted to a "home-made" approach, and succeeded in improving his mouth-opening by 18 mm. PMID- 16376000 TI - Solitary fibrous tumour of the buccal mucosa: case report and review of the literature. AB - Solitary fibrous tumours are spindle-cell neoplasms that usually occur in the pleura and peritoneum, and rarely involve the oral mucosa. We report a 30-year old man with a large solitary fibrous tumour on the buccal mucosa that resembled a salivary gland neoplasm. The lesion was excised and has not recurred. PMID- 16376001 TI - Maxillary reconstruction using rectus abdominis free flap and bone grafts. AB - We did total maxillectomies for cancer in 22 patients and reconstructed the face with rectus abdominis free flaps and bone grafts. The functional and aesthetic results have been satisfactory. The use of rectus abdominis free flap with nonvascularised bone grafts is a useful technique after excision of the maxilla and orbital floor. It permits a reliable, safe, one-stage reconstruction of complex three-dimensional anatomy with stable functional and aesthetic results. PMID- 16376002 TI - An unusual "sebaceous cyst". AB - A 41-year-old man was referred by his general practitioner to the general surgeons with an "infected sebaceous cyst" above the right medial canthus. "Cyst" was excised by the general surgeons. But this was followed by its recurrence after a number of weeks. A lateral skull radiograph showed the presence of a radio-opaque foreign body within the right orbit. PMID- 16376003 TI - Peripheral nerve compression neuropathy after fractures of the distal radius. AB - Sixty patients with unilateral distal radius fractures were managed conservatively. Clinical assessment included objective and subjective evaluations of the outcome of treatment. Radiographic evaluation included fracture classification using the AO/ASIF system and measurement of volar tilt, radial inclination and radial height shortening at the end of treatment. Clinical signs of carpal tunnel syndrome were confirmed electrodiagnostically in 12 patients (20%) during the recovery period. Each patient had electrodiagnostic studies of both upper limbs performed to confirm the diagnosis. The mean time between injury and the onset of symptoms was 10 months (range 6.5 weeks-27 months). A statistically significant correlation between the final clinical results of treatment and post-traumatic median nerve compression neuropathy was found, but the occurrence of the neuropathy correlated with neither fracture type nor with the final radiographic findings. No clinical signs of ulnar or radial nerve compression occurred in this study. PMID- 16376004 TI - Sonographically assisted percutaneous release of the a1 pulley: a new surgical technique for treating trigger digit. AB - This study introduces a sonographically assisted percutaneous technique for releasing trigger digits which provides direct visualization of the release and avoids the risks of incomplete release and injury to adjacent neurovascular structures associated with other percutaneous release techniques. The "safe zone" and an estimate of the size of the A1 pulley were determined in a separate cadaver study. We then used these landmarks in a prospective clinical study of 107 digits in 83 consecutive patients treated by this technique. During the follow-ups of between 9 and 15 months, we evaluated 104 digits in 80 patients. Pain was absent in 101 digits (97%) and considerably improved in the other three (3%). All mechanical problems had been resolved and none recurred during follow up. This technique allows the surgeon to see and monitor, precisely, the percutaneous division of the A1 pulley without open surgery and, therefore, to avoid the inherent risks of percutaneous and open surgical release. PMID- 16376005 TI - Selenium and other trace metals in fish inhabiting a fly ash stream: implications for regulatory tissue thresholds. AB - Bluegill sunfish (Lepomis macrochirus) and caddis flies (Trichoptera: Hydropsychidae) were collected from a stream receiving fly ash discharge and nearby reference streams to determine tissue levels of selenium (Se) and other metals, and compare these levels to published "no effect" thresholds. Stingy Run samples contained elevated levels of several metals. Mean Se concentrations in bullhead minnow whole body, bluegill whole body, bluegill ovary, and testes tissues were 44.6, 17.3, 32.5, and 37.1 microg/g (dry wt), respectively. These levels were 2-3 times higher than proposed toxic thresholds for fish whole body (7.9 microg/g) and ovary (17 microg/g). Although monitoring indicated a persistent bluegill population, some reproductive impairment may have occurred. Tissue residue data should be treated with caution because feral fish may accumulate several metals. In Stingy Run, persistence of a bluegill population may be explained by antagonistic interactions with other metals that were also elevated in the fish. PMID- 16376006 TI - Obesity, weight loss, and physician's advice. AB - Despite the increasing prevalence and economic costs of obesity in the USA, many physicians and other health care professionals do not advise their overweight and obese patients about weight loss. Using the 2001-2003 Behavioral Risk Factor Surveillance System data the purpose of our research is to investigate the relationship between individuals' receipt of physician's advice on weight loss and their tendency to eat fewer calories and fat or to use physical activity to lose weight. We find that physician's advice to lose weight has positive effects on both the probability of eating fewer calories and fat to lose weight and on the probability of using exercise to lose weight. PMID- 16376007 TI - Barriers to and attitudes towards promoting husbands' involvement in maternal health in Katmandu, Nepal. AB - Couple-friendly reproductive health services and male partner involvement in women's reproductive health have recently garnered considerable attention. Given the sensitive nature of gender roles and relations in many cultures, understanding the context of a particular setting, potential barriers, and attitudes towards a new intervention are necessary first steps in designing services that include men. In preparation for a male involvement in antenatal care intervention, this qualitative study specifically aims to: (a) understand the barriers to male involvement in maternal health and (b) explore men's, women's, and providers' attitudes towards the promotion of male involvement in antenatal care and maternal health. In-depth interviews were conducted with fourteen couples and eight maternal health care providers at a public maternity hospital in Katmandu, Nepal. Additionally, seventeen couples participated in focus group discussions. The most prominent barriers to male involvement in maternal health included low levels of knowledge, social stigma, shyness/embarrassment and job responsibilities. Though providers also foresaw some obstacles, primarily in the forms of hospital policy, manpower and space problems, providers unanimously felt the option of couples-friendly maternal health services would enhance the quality of care and understanding of health information given to pregnant women, echoing attitudes expressed by most pregnant women and their husbands. Accordingly, a major shift in hospital policy was seen as an important first step in introducing couple-friendly antenatal or delivery services. The predominantly favorable attitudes of pregnant women, husbands, and providers towards encouraging greater male involvement in maternal health in this study imply that the introduction of an option for such services would be both feasible and well accepted. PMID- 16376009 TI - Passive resistance increases differentially in various jaw displacement directions. AB - OBJECTIVES: In the present study, the passive resistance of the human jaw system was quantified in relation to the three-dimensional jaw displacement and the Posselt-envelope, using both in vivo measurements and computer simulation. METHODS: In eight subjects, the jaw was passively displaced with a step-wise increasing force in three orthogonal directions. Muscle relaxation was monitored using electromyography (EMG) with visual feedback. A biomechanical model of an average human system was used to examine the contributions of the jaw muscles. RESULTS: The largest excursion was found for the vertical direction. Protrusive and lateral directions were more restricted. In protrusive and lateral directions, the jaw could generally move beyond the Posselt-envelope. The stiffness of the jaw increased with proceeding jaw displacement in all directions. The stiffness was larger in the protrusive direction than in the vertical and lateral directions. The model's predictions of stiffness were comparable to the in vivo measurements. However, in protrusive direction, the maximum jaw displacement was larger than in vivo. The estimated passive muscle forces showed that vertical displacement was mainly restricted by the complete group of closing muscles, while protrusive and lateral jaw displacement was restricted by selective individual muscles. CONCLUSIONS: The human jaw system has larger motion range in the protrusive and lateral directions than can be exploited by active muscle use. Stiffness of jaw displacement is higher in the protrusive direction compared to the vertical and lateral directions. PMID- 16376008 TI - In vitro basal and metabolism-mediated cytotoxicity of flavonoids. AB - The purpose of this study was to compare the basal cytotoxicity and metabolism mediated cytotoxicity of kaempferol, quercetin and rutin. McCoy cells were exposed to various concentrations of the flavonols with and without the S9 system. The neutral red uptake assay was used to determine viability after 24 h at 35-37 degrees C. Dose-response curves were established for each flavonol in the presence and absence of external metabolizing systems. Kaempferol and quercetin were cytotoxic and provoked a dose-dependent decrease in cell viability, without the S9 system. The hepatic S9 microsomal fraction metabolized these compounds to less cytotoxic metabolites. In contrast, rutin at 500 microg/ml failed to produce any overt signs of toxicity in either assay. PMID- 16376010 TI - Implication of apoE isoforms in cholesterol metabolism by primary rat hippocampal neurons and astrocytes. AB - Apolipoprotein E (apoE) has been genetically linked to late-onset Alzheimer's disease. From the three common alleles (epsilon2, epsilon3 and epsilon4), epsilon4 has been suggested to promote amyloid beta (Ass) plaque fibrillation, one hallmark of Alzheimer's disease. It has been demonstrated that altered lipid content of hippocampal plasma membrane coincides with the disease. In this study, we show for the first time that the apoE dependent cholesterol metabolism in hippocampal neurons is higher than that of hippocampal astrocytes. Further, apoE bound cholesterol is highly incorporated in membranous compartments in hippocampal neurons, whereas hippocampal astrocytes show higher intracellular distribution. This is an effect that coincides with cell-type dependent difference of low density lipoprotein receptor (LDLR) family member expression. Hippocampal neurons express high levels of the LDLR related protein (LRP), whereas hippocampal astrocytes are highly positive for LDLR. We could also demonstrate an apoE isoform (apoE2, apoE3 and apoE4) dependent cholesterol uptake in both cells types. In hippocampal neurons, we could find a decreased apoE4 bound cholesterol uptake. In contrast, hippocampal astrocytes show decreased internalization of apoE2-bound cholesterol. In addition, lipidated apoE4 is little associated with neurites in hippocampal neurons in comparison to the other two isoforms. In contrary, hippocampal astrocytes show faint apoE2 immunocytostaining intensity. Data presented indicate that the role of apoE4 in cholesterol homeostasis and apolipoprotein cell association is more pronounced in hippocampal neurons, showing significant alterations compared to the other two isoforms, suggesting that hippocampal neurons are affected by apoE4 associated altered cholesterol metabolism compared to hippocampal astrocytes. PMID- 16376011 TI - Assessment of the rangelands of southwestern Santiago del Estero, Argentina, for grazing management and research. AB - Native rangelands of the southwest part of the province of Santiago del Estero, Argentina, are a key source of forage for cow-calf operations. The objectives of this study were to delineate the ecosystem units of the area, to describe the associated plant communities and to interpret the role that physical factors and disturbances such as fire and grazing have had in the changes of the structure of these plant communities. This information is needed for developing recommendations for grazing management, for prescribing appropriate improvement practices (e.g. shrub control, prescribed fire) and as guidelines for future research. The ecosystem was divided into smaller units using a hierarchical method, the categories of practical importance being 'range unit' and 'range site'. They represent the catchment and hillslope scale of the water runoff-runon phenomenon, respectively. Vegetation was sampled using a block and cluster sampling design, registering tree, shrub, forb and grass species frequency, and the standing aerial biomass of the herbaceous layer in a sampling unit=1 ha. Environmental data (topographic position, fire frequency, current and past use, and tree and shrub cover) were also registered for each sampling unit. Indirect ordination of sampling units classified according to range units and range sites, and correlation with environmental variables were performed using multidimensional scaling (MDS) as well as the vector fitting technique. Standing forage and stocking rate were estimated from biomass data. Results indicate that 'range site' is the ecosystem unit that should be considered for management purposes since it correlates well with plant communities: tall, hardwood forests are located on upland sites, woodlands are located on midland sites and savannas are located on lowland sites. Dense shrub thickets dominate in areas rated in poor condition, irrespective of range site. Disturbances such as fire and current and past use have a significant positive and negative correlation with range condition, respectively, suggesting that a state and transition model would explain vegetation dynamics better than the succession model. The estimated stocking rate in lowland sites in good condition was 2 ha UG(-1), while in upland sites in poor condition the stocking rate was 90 ha UG(-1). Active (fire, mechanical treatments) rather than passive (grazing management) methods should be used for range improvement in order to achieve the full potential of the ecosystem. PMID- 16376012 TI - The androgen receptor and prostate cancer invasion. AB - Recent evidence indicates that androgen-sensitive prostate cancer cells are characterized by a less pronounced malignant phenotype. We demonstrate that transfection with an androgen receptor (AR) expression vector of the androgen independent (AI) prostate cancer cell line PC3 decreases invasion and adhesion of these cells through modulation of alpha6beta4 integrin expression. Treatment of PC3-AR cells with the synthetic androgen R1881 further reduced invasion without modifying alpha6beta4 expression on the cell surface, suggesting interference with the invasion process in response to EGF by an alternative mechanism. We investigated EGF-induced auto-transphosphorylation of EGFR in both cell lines. We found that EGFR auto-transphosphorylation was reduced in PC3-AR cells and was further decreased by administration of androgens. Since auto-transphosphorylation regulates many different functions of EGFR, including docking of kinases, ubiquitination and internalization, we next investigated all these processes in PC3-AR cells. EGF-stimulated PI3K activity, a key signalling pathway for invasion of these cells, was decreased in PC3-AR cells and further reduced by treatment with R1881. Interestingly, EGFR-PI3K interaction was also disrupted in these cells. Furthermore, EGFR ubiquitination and internalization were found to be reduced in PC3-AR cells both in basal conditions and following treatment with androgens. According to recent findings, an endocytotic pathway may be important for EGFR signalling by controlling the specificity of the response. By using immunoconfocal fluorescent microscopy, we demonstrated that AR in PC3 cells is mainly located in cytoplasm and transmigrates in part to the nucleus following stimulation with androgens. Interestingly, immunoconfocal and immunoprecipitation experiments demonstrated also the occurrence of co-localization and interaction of AR with EGFR in PC3-AR cells and in another androgen-dependent PC cell line, LNCaP. We hypothesize a mechanism by which, through direct interaction with EGFR, the AR elicits a reduction of EGF-mediated signalling and confers a less malignant phenotype. PMID- 16376013 TI - CNS demyelination in autoimmune diseases. AB - Autoimmune diseases represent a diverse group of disorders that have generally of unknown etiology and poorly understood pathogenesis. They may be organ-specific or systemic, giving rise to overlapping syndromes; more than one autoimmune disease may occur in the same patient. Numerous case reports have documented that multiple sclerosis (MS) may be present concurrently with other autoimmune diseases, most commonly rheumatoid arthritis, autoimmune thyroid disease, type I diabetes mellitus and pernicious anemia. Case reports of disseminated encephalomyelitis (DEM) coincidental with other autoimmune diseases are rare. Many of systemic autoimmune diseases cause central nervous system (CNS) demyelination and are frequently then diagnosed as MS, whereas they often are instances of DEM, the result of vascular, granulomatous or postinfectious manifestations. We have reviewed 15 patients with autoimmune diseases and CNS demyelination in order to determine the nature of the demyelinating process. PMID- 16376014 TI - The differential diagnosis of acute transverse myelitis. AB - The clinical and paraclinical characteristics of acute transverse myelitis (ATM) were analyzed in 31 patients. In some patients there was clinical evidence of complete transection, in others of only partial lesions. Magnetic resonance imaging (MRI) in the acute phase in the first group was normal, but showed cord atrophy subsequently. It is probable that the clinical picture was due to parenchymatous neuronal lesions, analogous to those of axonal polyneuropathy. In the patients with incomplete transverse lesions, the most common finding was demyelination. In the patients with circumscribed demyelinating lesions, the symptoms and MRI were suggestive of clinically isolated syndromes (CIS) predictive of multiple sclerosis (MS). Extensive demyelination was indicative of acute disseminated encephalomyelitis (ADEM) due to hyperergic vasculopathy or various forms of chronic vasculitis. In two patients with variable clinical symptoms, a vascular malformation was the cause of the clinical presentation, and in one patient demyelination was due to the disc compression. PMID- 16376015 TI - Enlargement of the spinal cord: inflammation or neoplasm? AB - Intramedullary spinal tumours are uncommon lesions that can cause significant difficulties in the differential diagnosis between inflammatory diseases such as multiple sclerosis and acute disseminated encephalomyelitis, and vascular malformations or neoplasms. We report five cases in which the history and the clinical symptoms suggested an inflammatory process of the spinal cord but the MRI characteristics were those of neoplastic lesions. Both non-neoplastic and neoplastic intramedullary lesion may have very similar symptoms, and even CSF abnormalities, but in every one of our cases, a more detailed history and longer observation of the clinical course would have led to the correct diagnosis; in such problem cases, empirical treatment and a follow-up MRI after a month's observation would be a more prudent approach providing that the patient is not rapidly deteriorating. PMID- 16376016 TI - Emotion and movement: activation of defensive circuitry alters the magnitude of a sustained muscle contraction. AB - Understanding the emotion-movement relationship is crucial to the development of motor theory and movement rehabilitation recommendations for a wide range of diseases and injuries that involve motor impairment. Behaviorally, when movements are executed following exposure to emotional stimuli, evidence suggests that active defensive circuitry results in faster but more variable voluntary movements. However, each of the existing protocols has involved movement execution following the offset of anxiety or emotion eliciting stimuli. The specific aim of this study, therefore, was to determine whether the continued exposure to emotional stimuli would alter the magnitude and variability of a sustained motor contraction. During the presentation of pleasant, unpleasant, neutral, and blank images, participants (N=45) were instructed to respond to the onset of an auditory stimulus by initiating and then sustaining a maximal bimanual isometric contraction of the wrist and finger extensor muscles against two independent load cells (left/right limb). Corroborating previous evidence and supporting hypothesis 1, findings indicated that exposure to unpleasant images lead to an increase in mean force production. Variability of movement, however, did not vary as a function of affective context. These findings indicate that continued exposure to unpleasant stimuli magnifies the force production of a sustained voluntary movement, without sacrificing the variability of that contraction. Mechanism driven open and closed loop explanations are offered for these phenomena, implications are addressed, and future directions are discussed. PMID- 16376017 TI - Immunohistochemical localization of phospholipase D1 in the retina of pigs. AB - The expression of phospholipase D1 (PLD1) was examined in the retinas of pigs. Western blot analysis detected the expression of PLD1 in the retinas of 1-day-old piglets and showed that it was enhanced in the retinas of 2 years old adult pigs. Immunohistochemically, PLD1 was mainly immunostained in ganglion cell bodies in the ganglion cell layer, in some radial processes of Muller cells in the retinal layer and in the inner and outer segments of the rod and cone layer in newborn and adult pigs, but not in astrocytic bundles in nerve fiber layers. The immunoreactivity of PLD1 in the radial processes of Muller cells across the retinal layers was enhanced in adult pig retinas compared to those of newborn piglets. This was the first demonstration to show that PLD1 is constitutively expressed in the retina of pigs, implying that retinal PLD1 expression is enhanced in radial fibers of Muller cells with age. This finding suggests that PLD1 plays an important role in signal transduction of glial cells and neuronal cells in the retina. PMID- 16376018 TI - NO-mediated alterations in skeletal muscle nutritive blood flow and lactate metabolism in fibromyalgia. AB - The purpose of these investigations was to determine if differences exist in skeletal muscle nutritive blood flow and lactate metabolism in women with fibromyalgia (FM) compared to healthy women (HC); furthermore, to determine if differences in nitric oxide-mediated systems account for any detected alterations in blood flow and lactate metabolism and contribute to exertional fatigue in FM. FM (n = 8) and HC (n = 8) underwent a cycle ergometry test of aerobic capacity, a muscle biopsy for determination of nitric oxide synthase (eNOS, nNOS, iNOS) content, and microdialysis for investigation of muscle nutritive blood flow and lactate metabolism. During prolonged (3h) resting conditions, the ethanol outflow/inflow ratio (inversely related to blood flow) increased in FM over time compared to HC (P < 0.05). FM also exhibited a reduced nutritive blood flow response to aerobic exercise (P < 0.05). There was an increase in dialysate lactate in response to acetylcholine in FM, and to sodium nitroprusside in both groups, with a greater rise in dialysate lactate in FM (P < 0.05). The iNOS protein content was higher in FM and was negatively correlated with total exercise time (r(2) = 0.462, P < 0.05). IN CONCLUSION: (1) There is reduced nutritive flow response to aerobic exercise and reduced maximal exercise time in FM that might relate to higher iNOS protein content and contribute to exertional fatigue in FM; (2) The increased dialysate lactate in FM in response to stimulation of NOS or a nitric oxide donor suggest that FM may be more sensitive than HC to the suppressive effect of nitric oxide on oxidative phosphorylation. PMID- 16376019 TI - Assessment of burn area: can we be more objective? PMID- 16376020 TI - Functional outcome after burns: a review. AB - We conducted a Medline search (1966-11/2003) on empirical studies into the consequences of burns. The International Classification of Functioning, disabilities and health (ICF) was used to classify dimensions of functional outcome. We included 50 studies, reporting a wide spectrum of ICF-dimensions. The current state of knowledge on the functional outcome after burns was hard to summarise, due to the wide variety in study designs and outcome assessment methods. Some indications on the major functional problems after burns were gained. Problems in mental function were described in subgroups of patients, both in children/adolescents and adults. Restrictions in range of motion were observed in about one-fifth of burn patients, even 5 years after injury. Problems with appearance were reported often (up to 43%), even in patients with minor burns (14%). Problems with work were reported in 21-50% of the adult patients, with permanent incapacity for work in 1-5%. None of the publications gave sufficient information to fully estimate the functional consequences of burns. We recommend the development of a standard core set for measurement and reporting of functional outcome after burns. PMID- 16376021 TI - Massive squamous metaplasia of the thyroid gland-- report of three cases. AB - Three cases of massive squamous cell metaplasia in Hashimoto's thyroiditis are reported. The patients were two men and one woman aged 24, 52, and 55 years, respectively. In all three patients, the glandular parenchyma was replaced by hypocellular fibrous tissue with scattered chronic inflammatory infiltrate. Follicular cells were almost absent; the majority of residual epithelial cells formed squamous nests that were partly solid and partly cystic. There were three types of epithelial cells - squamous, basaloid, and follicular, with oncocytic differentiation. The squamous and basaloid cells showed strong positivity high molecular weight (HMW) cytokeratin, moderate to strong expression of galectin-3 (2/3), and nuclear expression of p63 protein (2/3). The staining pattern of p63 was identical to that of HMW, with predominant positivity at the periphery of cell nests. In one case, weak but unequivocal positivity of thyroid transcription factor-1 also was present. We believe that metaplasia was caused by Hashimoto's thyroiditis. The cases presented here are extremely rare, and only two convincing similar cases have been reported in the English literature so far. They may represent a diagnostic pitfall and should not be misdiagnosed as a malignancy, in particular as squamous cell or mucoepidermoid carcinoma. PMID- 16376022 TI - Hepatoprotective activity of Beta vulgaris against CCl4-induced hepatic injury in rats. AB - Ethanolic extract of Beta vulgaris roots given orally at doses of 1000, 2000 and 4000 mg/kg exhibited significant dose-dependent hepatoprotective activity against carbontetrachloride (CCl(4))-induced hepatotoxicity in rats. Hepatotoxicity and its prevention were assessed by serum markers viz. cholesterol, triglyceride, alanine amino transferase and alkaline phosphatase. PMID- 16376023 TI - Antidiabetic potential of Butea monosperma in rats. AB - The antihyperglycemic activity of the ethanolic extract of Butea monosperma (BMEE) was studied in glucose-loaded and alloxan-induced diabetic rats. Single dose treatment of BMEE (200 mg/kg, p.o.) significantly improved glucose tolerance and caused reduction in blood glucose level in alloxan-induced diabetic rats. Repeated oral treatment with BMEE (200 mg/kg/day) for 2 weeks significantly reduced blood glucose, serum cholesterol and improved HDL-cholesterol and albumin as compared to diabetic control group. PMID- 16376024 TI - Antidiarrhoeal activity of Cyperus rotundus. AB - The methanol extract of Cyperus rotundus rhizome, given orally at the doses of 250 and 500 mg/kg b.w., showed significant antidiarrhoeal activity in castor oil induced diarrhoea in mice. Among the fractions, tested at 250 mg/kg, the petroleum ether fraction (PEF) and residual methanol fraction (RMF) were found to retain the activity, the latter being more active as compared to the control. The ethyl acetate fraction (EAF) did not show any antidiarrhoeal activity. PMID- 16376025 TI - Characteristics of clinicians likely to refer clients to 12-Step programs versus a diversity of post-treatment options. AB - Most clients in substance abuse treatment are referred for continuing care. However, post-treatment services vary widely in their approaches to helping individuals achieve better substance use outcomes. This study examined the attitudes of outpatient treatment staff who refer clients exclusively to 12-Step groups (12-Step subgroup) and staff who refer clients both to 12-Step groups and to other continuing care options (Diversity subgroup) toward seven mutual-aid and professional psychosocial post-treatment options: Twelve-Step Programs (12-Step), Cognitive-Behavioral Therapy (CBT), Moderation Management (MM), Smart Recovery((R)) (SMART), Psychodynamic-oriented Therapy (PSY), Secular Organizations for Sobriety (SOS), and Women for Sobriety (WFS). A large percentage of clinicians lacked knowledge about the effectiveness of all alternatives to 12-Step programs with the exception of CBT. Clinicians in the 12 Step subgroup were more likely than those in the Diversity subgroup to be unfamiliar with alternatives to 12-Step programs and to believe less strongly in the effectiveness of CBT and PSY. A logistic regression found beliefs about CBT effectiveness and clinician preference for the 12-Step model to be related to the likelihood of referring exclusively to 12-Step groups. Findings suggest that clinicians could benefit from information and training on assessing and referring clients to various options for continuing care. PMID- 16376026 TI - Evolutionary fate of duplicate genes encoding aspartic proteinases. Nothepsin case study. AB - Gene duplication is considered an important evolutionary mechanism leading to new gene functions. According to the classical model, one gene copy arising from gene duplication retains the ancestral function, whilst the other becomes subject to directional selection for some novel functions. Hence, according to this model, long-term persistence of two paralogous genes is possible only with the acquisition of functional innovation. In the absence of neofunctionalization, one of the duplicate genes may be lost following accumulation of deleterious mutations, ultimately leading to the loss of function. Recently, new mechanisms have been proposed according to which both paralogs are maintained without apparent neofunctionalization. In this paper we describe the molecular evolution of the aspartic proteinase gene family, with particular regard for the nothepsin gene, a sex- and tissue-specific form of aspartic proteinase active in fish. The finding of nothepsin in a reptile is indicative of the presence of this gene in organisms other than fish. However, the failure to find any nothepsin-like gene in avian, murine and human genome suggests that the gene has been lost in certain lineages during evolution. At variance with piscine nothepsin expressed exclusively in female liver under the estrogens action, the reptilian counterpart lacks both tissue and sex specificity, as it is constitutively expressed in different tissues of male and female specimens. The expression of the nothepsin gene in fish and lizard is accompanied by the expression of a paralogous gene encoding for cathepsin D. Functional divergence analysis indicates that cathepsin D accumulated amino acid substitutions, whereas nothepsin retained most of the ancestral functions. Phylogenetic analysis shows a preponderance of replacement substitutions compared to silent substitutions in the branch leading to the cathepsin D clade, whilst nothepsin evolves under negative selection. To explain the loss of the nothepsin gene in certain lineages, we propose a model that takes into account the complementary degenerative mutations occurring in regulatory elements of the promoter regions of the two genes. According to this model, gene loss occurs whenever the two genes acquire the same expression pattern. The coexistence of cathepsin D and nothepsin is explained in terms of metabolic cooperation of the two enzymes. PMID- 16376027 TI - Divergent RNA editing frequencies in hornwort mitochondrial nad5 sequences. AB - Hornwort mitochondrial genomes have some of the highest rates of RNA editing among plants. Comparison of eleven partial mitochondrial nad5 genomic and cDNA sequences from diverse taxa of hornworts reveal 125 edited sites in only 1107 nt. No single sample exhibits more than half of these sites. Ten of the 11 hornwort taxa have between 35 and 54 edited sties each; whereas, the eleventh taxon, Leiosporoceros, which represents a potential sister taxa to all other hornworts, has only eight sites. Comparison of multiple cDNA sequences from several individuals reveals the presence of many immature transcripts showing the heterogonous nature of the progression of editing. Phylogenetic analyses of hornwort genomic and cDNAs sequences reveal that 65 of the 94 phylogenetically informative sites within the hornwort clade are edited positions. PMID- 16376028 TI - Identification of a constitutively active variant of LuxO that affects production of HA/protease and biofilm development in a non-O1, non-O139 Vibrio cholerae O110. AB - Pathogenesis of Vibrio cholerae depends on the concerted action of numerous virulence factors that includes a secreted hemagglutinin (HA) protease. Recent studies have evidenced that the expression of these virulence factors as well as the genes responsible for biofilm development is subject to control by quorum sensing in this organism. At low cell density, LuxO, the pivotal regulator of quorum-sensing circuit, has been shown to be phosphorylated at aspartate-47. Working in concert with sigma-54, LuxO-P activates the downstream repressor, which turned out to be four sRNAs [Lenz, D.H., Mok, K.C., Lilley, B.N., Kulkarni, R.V., Wingreen, N.S., Bassler, B.L., 2004. The small RNA chaperone Hfq and multiple small RNAs control quorum sensing in Vibrio harveyi and Vibrio cholerae. Cell 118, 69-82]. Subsequently, these sRNAs form complex with sRNA chaperone, Hfq. The Hfq-sRNA complex causes the destabilization of hapR mRNA transcript. HapR is a positive regulator of hapA that encodes HA/protease. At high cell density, dephosphorylation of LuxO impairs its function to activate the expression of sRNA, which in turn promotes HapR expression and causes protease production. It has been demonstrated that conversion of aspartate to glutamate (D47E) renders the LuxO molecule active without being phosphorylated. This variant of LuxO is referred as constitutively active LuxO or con-LuxO [Freeman, J.A., Bassler, B.L., 1999. A genetic analysis of the function of LuxO, a two component response regulator involved in quorum sensing in Vibrio harveyi. Mol Microbiol 31, 665-677]. Other than D47E, mutation at L104Q also develops con-LuxO [Vance, R.E., Zhu, J., Mekalanos, J.J., 2003. A constitutively active variant of the quorum-sensing regulator LuxO affects protease production and biofilm formation in Vibrio cholerae. Infect. Immun. 71, 2571-2576]. The purpose of this study was to investigate the cause of protease negative phenotype of a non-O1, non-O139 strain of V. cholerae O110. In the process of exploring the nature of the phenotype, a constitutively active variant of LuxO molecule was characterized which represses protease production and enhances biofilm formation by this strain. Unlike luxU, disruption of luxO restored the protease production, which showed the constitutively active nature of LuxO protein in this strain. PMID- 16376029 TI - Comparative genome analyses of nervous system-specific genes. AB - To elucidate the evolutionary process of the nervous system (NS) in metazoa, we examined the relationship between human genes specifically expressed in the NS (NS-specific genes) and the time of their evolutionary emergence. We obtained 255 human NS-specific genes from the gene expression data of the human full-length cDNA annotation invitational (H-invitational) database. To determine when these genes emerged for the first time during evolution, we searched for orthologues of the 255 NS-specific genes in 13 species (excluding human) by homology searches against their complete genome sequences. We found that 14% of the NS-specific orthologous genes had already emerged before the divergence between yeast and human. This finding suggests that a common ancestor, which should have no nervous system, already possessed a portion of the genes homologous to human NS-specific genes, implying that 14% of the NS-specific genes should have changed differentially their original functions during evolution. If this is the case, then the remaining 86% of the 255 NS-specific human genes have newly emerged during evolution. In particular, we found that the largest portion (24%) of the 255 NS-specific genes had emerged after divergence of urochordata and human but before divergence of fishes and human. These results suggest that the main cause of the NS evolution was the addition of new genes which took place most actively just before or at the evolutionary emergence of vertebrates. PMID- 16376030 TI - Managing the wildlife reservoir of Mycobacterium bovis: the Michigan, USA, experience. AB - Historical, social and economic factors combined to provide a focus where bovine tuberculosis has become established in free-ranging wildlife in northeastern lower Michigan. White-tailed deer, the primary reservoir and maintenance host of tuberculosis, are highly valued by the public, and particularly hunters, for cultural and economic reasons. Since 1995, significant progress has been made in defining and reducing the reservoir of tuberculosis in deer. As yet, no other wildlife species has been shown to play an epidemiologically important role in the disease cycle. The importance of deer and deer hunting to Michigan has uniquely shaped tuberculosis control policies, and poses ongoing challenges as wildlife managers strive to maintain momentum for broad control strategies, and develop focused strategies that are publicly acceptable. Even if momentum and funding can be maintained, tuberculosis will likely continue to be present for a decade or longer. Thus, cattle producers waiting for tuberculosis to be eradicated from wildlife to eliminate risks to their herds and markets face disappointment for the foreseeable future. Such unrealistic expectations also place Michigan's federal tuberculosis accreditation status at perpetual risk. Accredited free status is unlikely to be regained without accompanying changes in cattle management. In Michigan, management of tuberculosis has clearly demonstrated that social issues and public approval are likely to be the critical limiting factors in control. PMID- 16376031 TI - Acquisition of haemoglobin-bound iron by Histophilus somni. AB - Histophilus somni is an important pathogen of cattle and sheep. H. somni requires iron and can use ruminant transferrins as iron sources for growth. Here, we investigated the abilities of bovine (strains 649 and 2,336) and ovine (strains 9L and 3384Y) isolates of H. somni to acquire iron from haemoglobins. Using growth assays, the bovine isolates were shown to acquire iron from bovine haemoglobin, but not from ovine, porcine or human haemoglobins; the ovine isolates, however, failed to use any of these haemoglobins as iron sources for growth. In solid phase binding assays, the bovine isolates, grown under iron restricted conditions in the presence of bovine haemoglobin, bound not only bovine but also ovine and human haemoglobins. Competition binding assays indicated that all three haemoglobins were bound by the same receptor(s) and SDS PAGE of membrane fractions revealed that expression of haemoglobin-binding activity was associated with the production of an approximately 120-kDa outer membrane protein. PCR approaches allowed the amplification and sequencing of hgbA, and also hugX and hugZ homologues from strains 649, 9L and 3384Y. While hgbA of strain 649 was predicted to encode an HgbA precursor that is processed to yield a mature, 123.9-kDa haemoglobin-binding protein, the hgbA genes of strains 9L and 3384Y were predicted to give rise to truncated products. RT-PCR experiments revealed that in strain 649, hugX, hugZ and hgbA are co-transcribed and iron-regulated and additional sequencing suggested that in strain 2336, expression of HgbA is subject to phase variation involving a poly C tract within hgbA. PMID- 16376032 TI - Effect of intestinal fluid flux on ibuprofen absorption in the rat intestine. AB - Previously the apparent permeability coefficient (P(app)) of ibuprofen was observed to vary depending on the perfusion medium employed. The present work explores the possible contributions to these differences. Studies were undertaken using an in situ single pass rat gut technique. Lumenal drug concentrations and plasma drug levels were assayed by HPLC. Absorption rate constants (k(0)) were determined from fractions of drug unabsorbed from the intestineat steady state. Plasma data were fitted to a two compartment open model with zero-order input. Significant differences in net fluid flux were observed between the various buffered perfusion media, with fluxes varying from -0.044+/-0.006 ml min(-1) to +0.057+/-0.013 ml min(-1), the lower and negative values occurring for lower pH media and the larger positive values tending to occur with media of higher pH. A linear relationship was found between the P(app) of ibuprofen and net water flux (y=1.13+11.3x; r(2)=0.80). Apparent zero-order rate constants for ibuprofen appearance in plasma correlated well with absorption rate constants estimated from steady state lumenal drug concentration [k(0(gut))]. From the linear relationship between P(app) and fluid flux a normalized P(app) for ibuprofen (i.e. the P(app) in the absence of net fluid flux) of 1.1 x 10(-4)cms(-1) was determined Net lumenal fluid flux is dependent on perfusion medium composition and significantly alters ibuprofen absorption. The differences observed for P(app) were reflected in systemic drug absorption concentrations. The findings of these studies underline the importance of standardizing the osmolarity of experimental media used for the determination of intestinal permeability data. PMID- 16376033 TI - On the use of the Weibull function for the discernment of drug release mechanisms. AB - Previous findings from our group based on Monte Carlo simulations indicated that Fickian drug release from Euclidian or fractal matrices can be described with the Weibull function. In this study, the entire drug release kinetics of various published data and experimental data from commercial or prepared controlled release formulations of diltiazem and diclofenac are analyzed using the Weibull function. The exponent of time b of the Weibull function is linearly related to the exponent n of the power law derived from the analysis of the first 60% of the release curves. The value of the exponent b is an indicator of the mechanism of transport of a drug through the polymer matrix. Estimates for b< or =0.75 indicate Fickian diffusion in either fractal or Euclidian spaces while a combined mechanism (Fickian diffusion and Case II transport) is associated with b values in the range 0.75 or = 100g) are used. Hence, great amounts of the interfering elements are involved in the process, and therefore special emphasis is put on achieving a good commitment between adequate matrix elements removal and a minimization of the limit of detection. The performances of the method are analyzed in terms of conveniently defined figures of merit. The developed method is applied to several fallout level samples. In this way, the real performances and especially the real limitations of this method are shown. PMID- 16376090 TI - Some aspects involving the use of CdTe for finding end-point energies in diagnostic radiology. AB - We have considered the use of CdTe detectors and commercially available spectroscopy systems for the determination of the end-point energies of diagnostic X-ray devices. A simple technique based upon filtered spectra and simple fitting procedures was used to deduce end-point energies. We observed some unexpected detector-related effects such as significant shifts in the energy calibration at high count rates. When CdTe detectors operate at high counting rates drifts in the DC level, associated with collection rates of the positive charge (holes) can yield a corresponding shift in the spectrum towards lower energies. Only for relatively low counting rates and long detection periods, in excess of 10 min, can end-point energies be obtained with CdTe that reach levels of precision found in recent protocols. High rates also decrease energy resolution and lower the accuracy of recalibrations using characteristic X-rays from the target. PMID- 16376091 TI - Reevaluation of the emergency planning zone for nuclear power plants in Taiwan using MACCS2 code. AB - According to government regulations, the emergency planning zone (EPZ) of a nuclear power plant (NPP) must be designated before operation and reevaluated every 5 years. Corresponding emergency response planning (ERP) has to be made in advance to guarantee that all necessary resources are available under accidental releases of radioisotope. In this study, the EPZ for each of the three operating NPPs, Chinshan, Kuosheng, and Maanshan, in Taiwan was reevaluated using the MELCOR Accident Consequence Code System 2 (MACCS2) developed by Sandia National Laboratory. Meteorological data around the nuclear power plant were collected during 2003. The source term data including inventory, sensible heat content, and timing duration, were based on previous PRA information of each plant. The effective dose equivalent and thyroid dose together with the related individual risk and societal risk were calculated. By comparing the results to the protective action guide and related safety criteria, 1.5, 1.5, and 4.5km were estimated for Chinshan, Kuosheng, and Maanshan NPPs, respectively. We suggest that a radius of 5.0km is a reasonably conservative value of EPZ for each of the three operating NPPs in Taiwan. PMID- 16376092 TI - Intracoronary shunt reduces postoperative troponin leaks: a prospective randomized study. AB - OBJECTIVE: The purpose of this study was to evaluate whether intracoronary shunt usage reduced the myocardial damage on the basis of the cardiac markers when compared with the shuntless anastomosis in off-pump coronary artery bypass grafting (OPCABG) surgery of isolated left anterior descending artery lesions. METHODS: Forty patients who had stable angina with isolated left anterior descending (LAD) coronary artery lesion undergoing OPCABG surgery were randomized into two groups. Shunt group consisted of 20 patients who had OPCABG using intracoronary shunt, whereas the shuntless group consisted of 20 patients who underwent OPCABG without using intracoronary shunt. Cardiac troponin I, CK, and CK-MB before and 24h after the surgery were assessed in the groups. RESULTS: There were no deaths in the study. The two groups were similar with respect to sex and age. Duration of LIMA-LAD anastomosis was significantly higher in the shunt group (p=0.01). There was no significant difference between the groups concerning the preoperative and postoperative CK and CK-MB levels. The preoperative troponin I levels of the groups were not different (p=0.238; NS), whereas postoperative levels of this marker was significantly higher in the shuntless group (p=0.003). CONCLUSION: Intracoronary shunt reduced the postoperative troponin I levels significantly, so it may be indicated in the patients who are thought to be susceptible to transient ischemia. PMID- 16376093 TI - Prognostic value of chronic obstructive pulmonary disease in coronary artery bypass grafting. AB - OBJECTIVE: Coronary and chronic lung diseases have become a common association. This comorbidity has been generically considered by most of the operative risk scores, but its functional severity has seldom been addressed by these models. Our objective was to analyze its prognostic relevance considering preoperative pulmonary function parameters. METHODS: All patients undergoing CABG from May 1993 to December 2004 have been reviewed. One thousand four hundred and twelve patients with preoperative pulmonary function test were finally included in the study. Obstructive lung disease was defined when FEV1/FVC<0.7. In-hospital mortality and complication rate related to chronic obstructive pulmonary disease and its degree of severity (FEV1%) were assessed. Logistic regression analysis was used to determine independent predictors of mortality. RESULTS: A pathologic preoperative pulmonary function test was found in 39% of patients: obstructive in 26% (FEV1/FVC<0.7), restrictive in 9% and combined obstructive-restrictive in 4%. In-hospital mortality was higher in patients with abnormal test: 6.5% versus 0.9% (p<0.001). Mortality was clearly related with the severity of lung disease: 0.9% in patients with FEV1: >80%, 0.4% in FEV1: 60-80%, 10.8% in FEV1: 40-59% and 54% in FEV1: <40%. In the latter group, other intercurrent prognostic factors were observed. Patients with FEV1<60% had higher mortality than those with FEV1>60%: 24.6% versus 1.4% (p<0.001). Chronic obstructive lung disease was not an independent predictor of mortality but FEV1< or =60% was significantly associated with death. CONCLUSIONS: This study on chronic lung comorbidity in CABG patients shows that this association can be of deleterious prognostic value but this effect is directly related to the degree of functional severity. Preoperative FEV1<60% must be considered as a primary prognostic factor in patients undergoing CABG procedures. PMID- 16376094 TI - Extended pulmonary resections of advanced thoracic malignancies with support of cardiopulmonary bypass. AB - OBJECTIVE: Complete resection of advanced pulmonary malignancies infiltrating the heart or the great vessels may require the application of cardiopulmonary bypass (CPB). Extracorporal circulation, however, is known to cause lung injury and may be harmful especially in pneumonectomies. METHODS: Over a period of 10 years extended pulmonary resections requiring cardiopulmonary bypass were analyzed in a retrospective study. RESULTS: From August 1993 to August 2003, 13 patients underwent an extended pulmonary resection for curative indications, requiring support by CPB. Underlying diseases were sarcomas (n=8), non-small cell lung carcinomas (n=3), and others (n=2). Pneumonectomies were performed in nine and lobectomies in four cases. In the majority of cases, several cardiac structures, predominantly the left atrium (n=9), were affected. In four patients (31%), the indication for a CPB-supported procedure was not electively planned, but made intraoperatively. Complete en-bloc resection (R0) was achieved in 12 of 13 cases (92%). The 30-day mortality rate was 15% (n=2). Major complications observed were acute lung injury (n=4), right heart failure (n=1), and multi-organ failure (n=1). The cumulative survival at 1, 3, and 5 years in patients presenting with sarcomas was 62.5% compared to 33%, 0%, and 0%, respectively, in patients with non-small cell carcinoma (n=3). CONCLUSIONS: Our results encourage the application of CPB in extended pulmonary resections to achieve complete resections. In carefully selected patients, especially those with sarcomas, the radical surgical procedure associated with increased pulmonary complications allows for significantly prolonged survival and quality of life. PMID- 16376095 TI - Video-assisted thoracoscopic surgery for primary spontaneous pneumothorax: clinicopathological correlation. AB - OBJECTIVE: To compare the identifiable pulmonary abnormalities during thoracoscopy with the histological findings in patients requiring surgical intervention for recurrent or persistent primary spontaneous pneumothorax (PSP) and correlate these with the postoperative events. METHODS: From January 1999 to December 2002, 94 consecutive patients underwent video-assisted thoracoscopic wedge excision and apical pleurectomy for PSP. Vanderschueren's classification was used for macroscopic staging and histological observation for microscopic features. Clinical data of these patients and the outcome of surgery were described. RESULTS: All patients were successfully treated using video-assisted thoracoscopic technique. Recurrent pneumothorax was the most frequent indication for surgery, occurring in 60 cases. The method of management was stapling of an identified bleb or apex of the upper lobe and apical pleurectomy. In 67 cases (71%), clear bullae were found in types III and IV. In 15 cases (16%), type II pleuropulmonary adhesions were identified and in 12 (13%) cases thoracoscopy failed to reveal any abnormality (type I). The actual site of air leakage could be located during thoracoscopy in 24 (26%) patients. Histologically, 74 patients had subpleural bullae/blebs formation and 20 had emphysema without bullae. Fifty three patients had cellular infiltration and 82 had pleural fibrosis. In the microscopic examination, the actual site of air leakage could be located at the site of subpleural blebs or bullae in 15 patients and elsewhere at the lung surface in five other patients. Postoperative prolonged air leak occurred in 4 out of 12 patients in type I and in two of the remaining patients, p=0.001. Mean follow-up is 48 months (range, 30-60 months) for all patients. Pneumothorax recurred in three patients (3.1%). Two patients from type I (16.6%) and one patient from the other types (1.2%) had recurrence (p=0.01). CONCLUSIONS: Video assisted thoracoscopic stapling of an identified bleb or apex of the upper lobe and apical pleurectomy represents the standard treatment for the majority of recurrent or persistent PSP. Most patients with surgically treated PSP have subpleural blebs or bullae or isolated emphysema. In type I cases, simple apical excision and apical pleurectomy are not sufficient and perhaps additional talc poudrage might be indicated. PMID- 16376096 TI - Performance at 10 years of the CarboMedics "Top-Hat" valve. Postclamping time is a predictor of mortality. AB - OBJECTIVE: The CarboMedics "Top-Hat" supraannular prosthesis was designed to permit the implantation of a larger prosthesis. We evaluated the outcome at 10 years in patients with this prosthesis. METHODS: Between June 1993 and May 2001, 269 patients (average age, 63.9 years) received a CarboMedics "Top-Hat" supraannular aortic prosthesis. Primary valve replacement was performed on 203 patients (75.5%) and repeat valve replacement on 66 (24.5%). The duration of myocardial ischemia was 70.2+/-31.4 min, cardiopulmonary bypass 96.1+/-48.3 min, and postclamping time (time between release of aortic clamp and the end of extracorporeal circulation) 22.1+/-41.3 min. The mean follow-up was 82.3+/-17.8 months. Follow-up was 97.6% complete. RESULTS: The hospital mortality was 5.9%. It was 1% when the duration of postclamping time was <15 min, 2.8% between 15 and 29 min, 13.2% between 30 and 44 min, and 26.9% >44 min. In the multivariate analysis, postclamping time, urgent surgery, and body mass index were statistically significant risk factors for hospital mortality. The late mortality was 17.1%. Cardiac-related mortality showed a linearized rate of 18.1% per 1000 patients-year. The Kaplan-Meier estimates for cardiac-related mortality was 75.0% at 10 years. Postclamping time, aortic valve gradient, age over 70 years, and BMI were statistically significant risk factors for cardiac-related late mortality. The incidence of paravalvular leak in the "Top-Hat" aortic prosthesis was 1.7% per 1000 patients-year. CONCLUSIONS: Using the CarboMedics supraannular prosthesis allows implantation of a larger prosthesis without increasing valve related complications. Postclamping time appears as a strong predictor of both hospital mortality and late cardiac-related death. PMID- 16376097 TI - Infected pulmonary artery aneurysms: CT imaging findings. PMID- 16376098 TI - Successful heart transplantation in Becker's muscular dystrophy. PMID- 16376099 TI - Surgery for type A aortic dissection in pregnant patients with Marfan syndrome. PMID- 16376100 TI - Use of an electrothermal bipolar tissue sealing system in lung surgery. AB - OBJECTIVE: We evaluated the validity of an electrothermal bipolar tissue sealing system (LigaSure, Valleylab Inc., USA) in lung surgery. METHODS: Our study was divided into two phases. EXPERIMENTAL: We performed sutures of pulmonary vessels and bronchi and lung wedge resections by LigaSure in 28 lungs of adult pigs; subsequently, we quantitated the sealing capacity of the system detecting the burst pressure for each anatomical structure. Clinical: LigaSure was used in 36 patients undergoing lung surgery. We performed 23 thoracotomic procedures in 16 patients (19 wedge resections, 2 segmentectomies, and 2 fissure separations), and 20 thoracoscopic procedures (13 wedge resections, 5 bullectomies, and 2 adherence dissections). RESULTS: EXPERIMENTAL: Bronchi and vessels were divided into seven groups (diameter: 1-7 mm); 10 burst pressure measurements for each group were performed. A total of 84 wedge resections were performed; lung specimens were divided into seven groups (weight: 0.2-1.4 g). The percentage of bronchial sutures resistant to the pneumatic critical pressure (60 mmHg) was 100% in the 1 mm and 2-mm groups. No bronchi with 6-mm or 7-mm diameter reached the critical pressure. All pulmonary vessel sutures were resistant to the critical hydrostatic pressure (150 mmHg). The average burst pressure of wedge resection margins was higher than the critical pressure, and the percentage of suture margins resistant to the critical pressure decreased from 95% (0.2-g group) to 68% (1.4-g group). Histology confirmed the sealing of vessels, with a mean depth of thermal injury limited to 1.1mm. Clinical: In all patients, hemostasis obtained by LigaSure was effective, with minimal perioperative bleeding. The mean operating time was 77.2 min (range: 60-97) for thoracotomies and 60.3 min (range: 46-80) for thoracoscopies. The mean drainage duration was 3.1 days (range: 1-8). Two patients had prolonged air leaks (>7 days). The mean postoperative stay was 7.3 days (range: 5-13) for thoracotomies and 4.6 days (range: 1-6) for thoracoscopies. CONCLUSIONS: Use of LigaSure in lung surgery appears feasible and easy. It provides satisfactory hemostasis and air-leak prevention; results are comparable to those of stapling devices, but this system seems to have a better benefit/cost ratio. Larger series are needed to confirm these data. PMID- 16376101 TI - Suppression of anti-cancer immunity by regulatory T cells: back to the future. AB - Suppressor/regulatory T cells were first shown to have an impact on cancer progression in experimental tumor models during the 1970s. However, the lack of specific markers hindered mechanistic investigations, and skepticism grew in the scientific community due to variability in cell populations and reported functions. The identification of regulatory CD4(+)CD25(+) T cells has generated a great deal of renewed interest in cells that have immune regulatory properties. This article will provide a brief historical review of suppressor T cells and cancer, experimental and clinical evidence that CD4(+)CD25(+) natural regulatory T cells play a role in cancer progression, and briefly discuss current strategies to inhibit these cells in an effort to enhance cancer immunotherapy. PMID- 16376102 TI - Regulatory T cells in immune surveillance and treatment of cancer. AB - Naturally occurring CD25(+)CD4(+) regulatory T cells (T(R) cells), which specifically express the transcription factor Foxp3, engage in the maintenance of immunological self-tolerance and suppressive control of aberrant or excessive immune responses to foreign antigens. They may, on the other hand, impede immune surveillance against cancer and hamper the development of effective immunity to autologous tumor cells. Indeed, natural T(R) cells have been observed to predominantly infiltrate tumor masses especially in the early phase of tumor progression. Depletion of natural T(R) cells by removing CD25(+) T cells prior to tumor challenge is therefore able to provoke effective tumor immunity in animals. Furthermore, attenuation of T(R) cell-mediated suppression in on-going anti-tumor immune responses, for example by altering signaling through CTLA-4 or GITR expressed by natural T(R) cells, can enhance the responses and thereby eradicate advanced cancers. A combination of depletion or attenuation of T(R) cells and concomitant stimulation of effector T cells, systemically or locally in tumors, may be a feasible immunotherapy for cancer. PMID- 16376104 TI - Expression and characterization of recombinant tyramine beta-monooxygenase from Drosophila: a monomeric copper-containing hydroxylase. AB - We report here the development of a robust recombinant expression system for Drosophila melanogaster tyramine beta-monooxygenase (TbetaM), the insect analog of mammalian dopamine beta-monooxygenase. Recombinant TbetaM is rapidly purified from the host cell media in three chromatographic steps. The expression system produces approximately 3-10 mg of highly purified, active protein per liter of culture. Recombinant TbetaM requires copper for activity and has a typical type 2 copper EPR spectrum. While TbetaM efficiently hydroxylates the aliphatic carbon of phenolic amines such as tyramine (the physiological substrate) and dopamine, phenethylamine is a poor substrate. TbetaM is most likely a monomer under physiological conditions, although under conditions of high pH and low ionic strength the dimeric form predominates. The lower oligomeric state of TbetaM may provide an advantage for structural studies over DbetaM, which exists as a mixture of dimer and tetramer. PMID- 16376103 TI - Inhibition of sphingolipid synthesis affects kinetics but not fidelity of L1/NgCAM transport along direct but not transcytotic axonal pathways. AB - Glycosphingolipids are constituents of lipid rafts which might function in sorting apical and axonal cargoes in the trans-Golgi network. In fact, two GPI linked proteins, Thy1 and PrPC, require lipid raft lipids for sorting to the axon. It was previously shown that inhibition of glycosphingolipid synthesis by FumonisinB1 (FB1) impairs axon outgrowth but not axon specification, leading to the hypothesis that formation of axonally-targeted vesicles is coupled to sphingolipid synthesis. Since the axonal cell adhesion molecule L1/NgCAM can partition into membrane rafts biochemically, we asked whether correct targeting to the axon is FB1-sensitive, similarly to GPI-linked proteins. We previously showed that cultured hippocampal neurons use more than one trafficking pathway to the axon: a transcytotic pathway and a direct pathway. We show here that reducing raft lipid levels does not disrupt axonal targeting of L1/NgCAM along either pathway. Unexpectedly, FB1 selectively slowed the kinetics of surface expression of a truncated NgCAM using the direct pathway, but not of NgCAM using the transcytotic pathway. Therefore, the formation and/or transport of a subset of axonally-targeted vesicles are coupled to sphingolipid synthesis. Our results yield a mechanism for the axon outgrowth defect observed in FB1. PMID- 16376105 TI - Large-scale purification and characterization of human parathyroid hormone-1 receptor stably expressed in HEK293S GnTI- cells. AB - Human parathyroid hormone-1 receptor (hPTHR1) belongs to class II of the G protein-coupled receptor (GPCR) family, whose members all contain a seven transmembrane helix domain. The receptor regulates bone metabolism through interactions with its ligand, human parathyroid hormone (hPTH). For structural studies of the hPTHR1/hPTH complex, we constructed a mammalian cell line to stably express recombinant hPTHR1 in large-scale. The receptor was solubilized with dodecyl maltoside and purified with affinity chromatography. The purified receptor displayed restricted N-glycosylation as expected. Functionality was demonstrated: the hPTHR1 retained affinity for bPTH-(1-34) and specifically cross linked to a radioiodinated bPTH-(1-34) analog. This work describes an approach for preparing milligram-scale quantities of receptor for elucidation of the structural biology of this seven-transmembrane GPCR. PMID- 16376106 TI - Molecular insight into pseudolysin inhibition using the MM-PBSA and LIE methods. AB - Pseudolysin, the extracellullar elastase of Pseudomonas aeruginosa (EC: 3.4.24.26) plays an important role in the pathogenesis of P. aeruginosa infections. In the present study, molecular dynamics simulations and theoretical affinity predictions were used to gain molecular insight into pseudolysin inhibition. Four low molecular weight inhibitors were docked at their putative binding sites and molecular dynamics (MD) simulations were performed for 5.0 ns, and the free energy of binding was calculated by the linear interaction energy method. The number and the contact surface area of stabilizing hydrophobic, aromatic, and hydrogen bonding interactions appears to reflect the affinity differences between the inhibitors. The proteinaceous inhibitor, Streptomyces metalloproteinase inhibitor (SMPI) was docked in three different binding positions and MD simulations were performed for 3.0 ns. The MD trajectories were used for molecular mechanics-Poisson-Boltzmann surface area analysis of the three binding positions. Computational alanine scanning of the average pseudolysin-SMPI complexes after MD revealed residues at the pseudolysin-SMPI interface giving the main contribution to the free energy of binding. The calculations indicated that SMPI interacts with pseudolysin via the rigid active site loop, but that also contact sites outside this loop contribute significantly to the free energy of association. PMID- 16376107 TI - Analysis and validation of automated skull stripping tools: a validation study based on 296 MR images from the Honolulu Asia aging study. AB - As population-based epidemiologic studies may acquire images from thousands of subjects, automated image post-processing is needed. However, error in these methods may be biased and related to subject characteristics relevant to the research question. Here, we compare two automated methods of brain extraction against manually segmented images and evaluate whether method accuracy is associated with subject demographic and health characteristics. MRI data (n = 296) are from the Honolulu Asia Aging Study, a population-based study of elderly Japanese-American men. The intracranial space was manually outlined on the axial proton density sequence by a single operator. The brain was extracted automatically using BET (Brain Extraction Tool) and BSE (Brain Surface Extractor) on axial proton density images. Total intracranial volume was calculated for the manually segmented images (ticvM), the BET segmented images (ticvBET) and the BSE segmented images (ticvBSE). Mean ticvBSE was closer to that of ticvM, but ticvBET was more highly correlated with ticvM than ticvBSE. BSE had significant over (positive error) and underestimated (negative error) ticv, but net error was relatively low. BET had large positive and very low negative error. Method accuracy, measured in percent positive and negative error, varied slightly with age, head circumference, presence of the apolipoprotein eepsilon4 polymorphism, subcortical and cortical infracts and enlarged ventricles. This epidemiologic approach to the assessment of potential bias in image post-processing tasks shows both skull-stripping programs performed well in this large image dataset when compared to manually segmented images. Although method accuracy was statistically associated with some subject characteristics, the extent of the misclassification (in terms of percent of brain volume) was small. PMID- 16376108 TI - Investigating direct detection of axon firing in the adult human optic nerve using MRI. AB - The aim of this study was to directly detect spectral components of the magnetic fields of ionic currents caused by firing of the axons in the optic nerve in response to visual strobe stimulation. The magnetic field parallel to the main B0 field can potentially alter the local phase and magnitude of the MR signal which can cause signal loss due to intravoxel dephasing. Measured frequency spectra showed evidence of the strobe stimulus localized to regions containing the optic nerve, not thought to be due to motion artifacts, in 30 out of 52 experiments in 5 adult human subjects. The effect was (0.15 +/- 0.05)% of the mean magnitude equilibrium signal from the voxel in the frequency range 0.7-3.3 Hz, corresponding to an estimated field of (1.2 +/- 0.4) nT, at an echo time of TE = 32.4 ms using a 1.5 T MRI scanner. Only 1 of 12 phase image experiments showed effects. These findings provide preliminary evidence for direct detection of axonal firing in the optic nerve. PMID- 16376109 TI - Studies of morphological and molecular phylogenetic divergence in spiders (Araneae: Homalonychus) from the American southwest, including divergence along the Baja California Peninsula. AB - Comparative phylogenetic and phylogeographic analyses have revealed a pervasive midpeninsular divergence in the mitochondrial genealogies of numerous vertebrate taxa distributed on the Baja California Peninsula. In this study, we extend the investigation of regional vicariance in Baja California to an arthropod taxon by examining patterns of phylogenetic and morphological divergence in the spider genus Homalonychus (Araneae, Homalonychidae). We analyzed data from two mtDNA genes (16S rRNA and NADH dehydrogenase subunit (1) and a nuclear gene (28S rRNA) using maximum parsimony and Bayesian phylogenetic analyses, and also conducted geometric morphometric analyses employing landmark data on male and female genitalia. Genes and morphology both reveal a deep split across the Colorado River and Gulf of California, separating Homalonychus selenopoides on the east side of river from its congener Homalonychus theologus on the west side of the river, including the Baja California Peninsula. Along the north-south axis of the Baja Peninsula, an apparently more recent midpeninsular phylogenetic break is evident within H. theologus in the mitochondrial genome and in female genitalia. However, there is no measurable divergence between northern and southern populations in either nuclear DNA or male genitalia. We suggest that this discordance between datasets reflects either a difference in rates of evolution between male versus female systems, or that male-based nuclear gene flow is obscuring a phylogenetic split that is fixed in the female-based systems. Our findings provide additional support for a midpeninsular Baja divergence event, although the timing and geological evidence for such an event remain elusive. PMID- 16376110 TI - Phylogeography of the Galapagos hawk (Buteo galapagoensis): a recent arrival to the Galapagos Islands. AB - Galapagos hawks (Buteo galapagoensis) are one of the most inbred bird species in the world, living in small, isolated island populations. We used mitochondrial sequence and nuclear minisatellite data to describe relationships among Galapagos hawk populations and their colonization history. We sampled 10 populations (encompassing the entire current species range of nine islands and one extirpated population), as well as the Galapagos hawk's closest mainland relative, the Swainson's hawk (B. swainsoni). There was little sequence divergence between Galapagos and Swainson's hawks (only 0.42% over almost 3kb of data), indicating that the hawks colonized Galapagos very recently, likely less than 300,000 years ago, making them the most recent arrivals of the studied taxa. There were only seven, closely related Galapagos hawk haplotypes, with most populations being monomorphic. The mitochondrial and minisatellite data together indicated a general pattern of rapid population expansion followed by genetic isolation of hawk breeding populations. The recent arrival, genetic isolation, and phenotypic differentiation among populations suggest that the Galapagos hawk, a rather new species itself, is in the earliest stages of further divergence. PMID- 16376111 TI - NF-kappaBp65-specific siRNA inhibits expression of genes of COX-2, NOS-2 and MMP 9 in rat IL-1beta-induced and TNF-alpha-induced chondrocytes. AB - OBJECTIVE: Small interfering RNA (siRNA) triggers RNA interference in mammalian somatic cells. Nuclear factor kappaB (NF-kappaB) is a transcription factor that is implicated in inflammation and immune activation. This study was to use NF kappaBp65-specific siRNA to inhibit the expression of genes of cyclooxygenase-2 (COX-2), nitric oxide synthase-2 (NOS-2) and matrix metalloproteinase-9 (MMP-9), which is paralleled with the initiation and progression of cartilage lesions in osteoarthritis (OA) model, in induced chondrocytes, and therefore to explore a new gene therapy for OA. METHODS: Western blot and reverse transcriptase polymerase chain reaction (RT-PCR) were performed to optimize the silencing effects of NF-kappaBp65-specific siRNA in cultured rat chondrocytes, and then to determine the expression of COX-2, NOS-2 and MMP-9 in induced chondrocytes. The activation of NF-kappaB was determined by electrophoretic mobility shift assay (EMSA). Western blot and RT-PCR were subjected to densitometric analysis and then band intensities were also determined. RESULTS: The NF-kappaBp65-specific siRNA inhibited the expression of NF-kappaBp65 and activation of NF-kappaB, reducing significantly the expression of COX-2, NOS-2 and MMP-9 induced by interleukin 1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in cultured chondrocytes. CONCLUSIONS: NF-kappaBp65-specific siRNA can inhibit the expression of COX-2, NOS-2 and MMP-9 in IL-1beta-induced and TNF-alpha-induced chondrocytes. This suggests that NF-kappaBp65-specific siRNA has potential to be a useful, preventive and therapeutic agent for OA at early stages. PMID- 16376112 TI - Presenilin-1 controls the growth and differentiation of endothelial progenitor cells through its beta-catenin-binding region. AB - Presenilin-1 (PS1) is a gene responsible for the development of early-onset familial Alzheimer's disease. Targeted disruption of the PS1 gene in mice suggested that PS1 might be involved in angiogenesis. We have used an in vitro embryonic stem (ES) cell culture system to prepare endothelial progenitor cells (EPC) lacking PS1 and investigated the roles of PS1 in endothelial cell lineage. With this system, Flk-1+ E-cadherin- EPC were generated from PS1-deficient ES cells, and the EPC lacking PS1 as well as wild-type EPC grew to form VE-cadherin+ endothelial colonies supported by a layer of OP9 stromal cells. Although the endothelial colonies from PS1-deficient EPC showed morphology similar to those from wild-type EPC, the PS1-deficient EPC formed a large number of the colonies compared to wild-type EPC. The enhanced colony-forming ability of PS1-deficient EPC was attenuated by the inductions of wild-type human PS1. To differentiate multiple activities of PS1 for colony-forming ability, we used two types of human PS1 mutants: one (hPS1D257A) with the aspartate to alanine mutation at residue 257 that impairs the proteolytic activity of PS1, and the other (hPS1Deltacat) deleting amino acids 340-371 of the cytosolic loop sequence essential for beta catenin binding. hPS1D257A showed activity to regulate the colony-forming ability of PS1-deficient EPC, while hPS1Deltacat failed to exhibit this activity. These results suggest that PS1 regulates the growth and differentiation of endothelial progenitor cells through its beta-catenin-binding region and that the defect of PS1 function in endothelial cell lineage could contribute to the induction of vascular pathology. PMID- 16376113 TI - Osmotically unresponsive water fraction on proteins: non-ideal osmotic pressure of bovine serum albumin as a function of pH and salt concentration. AB - How much does protein-associated water differ in colligative properties (freezing point, boiling point, vapor pressure and osmotic behavior) from pure bulk water? This question was approached by studying the globular protein bovine serum albumin (BSA), using changes in pH and salt concentration to alter its native structural conformation and state of aggregation. BSA osmotic pressure was investigated experimentally and analyzed using the molecular model of Fullerton et al. [Biochem Cell Biol 1992;70(12):1325]. Analysis yielded both the extent of osmotically unresponsive water (OUW) and the effective molecular weight values of the membrane-impermeable BSA solute. Manipulation of BSA conformation and aggregation by membrane-penetrating cosolutes show that alterations in pH and salt concentration change the amount of bulk water that escapes into BSA from a minimum of 1.4 to a maximum of 11.7 g water per g dry mass BSA. PMID- 16376114 TI - Anion-exchange blocker enhances cytoplasmic vacuole formation and cell death in serum-deprived mouse kidney epithelial cells in mice. AB - An anion exchange blocker, DIDS, exhibits anti-apoptotic activity in response to several apoptotic stimuli, but has an opposite effect when apoptosis is induced by serum deprivation. After adding DIDS, serum-deprived MCT cells exhibited vacuole formation in their cytoplasm and underwent cell death. Caspase activity increased with the addition of DIDS to serum-deprived MCT cells, but Z-Asp-CH2 DCB, a caspase inhibitor, did not inhibit cell death in DIDS-treated, serum deprived MCT cells. Cathepsin is considered to be important for vacuole formation and cell lysis, and pepstatin A, a cathepsin D inhibitor, partially inhibited vacuole formation in DIDS-treated, serum-deprived MCT cells, although caspase activation was not inhibited. 3-Methyladenin inhibited vacuole formation and cell death in DIDS-treated, serum-deprived MCT cells. These results suggest that DIDS treated serum-deprived MCT cells undergo autophagy, not apoptosis. PMID- 16376116 TI - Mid-term results of conservative, conventional and endovascular treatment for acute traumatic aortic lesions. AB - BACKGROUND: To analyze our results after conservative, conventional and endovascular treatment for acute traumatic aortic lesions during the last decade. METHODS: From June 1993 to September 2004, a total of 19 patients with traumatic aortic lesions were referred to our department. All patients sustained injuries from blunt deceleration trauma. In hemodynamically stable patients, initial evaluation was by multi-slice CT scan. The diagnosis of traumatic aortic injury was confirmed and an individual treatment strategy was determined. In hemodynamically unstable patients, emergency thoracotomy was performed. RESULTS: An emergency thoracotomy was performed in seven (37%) patients. Mortality in this group was 100%. In the remaining group of 12 (63%) patients without hemodynamic instability at time of admission, in-hospital mortality was 0%. Treatment was surgical in five patients (26%), endovascular in five (26%) and conservative in two patients (11%). Mean follow-up was 63 months (5-108 months). No patient died during follow-up. In patients treated by endovascular stent-graft placement no signs of endoleaks could be detected. CONCLUSIONS: Hemodynamic stability and an individual treatment strategy are prerequisites for survival of acute traumatic aortic lesions. Endovascular stent-graft placement has emerged as an innovative and minimally invasive therapeutic option in this polytraumatic high-risk patient cohort. PMID- 16376115 TI - Cognitive disruption and altered hippocampus synaptic function in Reelin haploinsufficient mice. AB - The heterozygote reeler mouse (HRM) shows many neuroanatomical and biochemical features that are also present in some human cognitive disorders, such as schizophrenia. In the present study, hippocampal dependent plasticity and cognitive function of the HRM were characterized in detail in an attempt to reveal phenotypic functional differences that result from Reelin haploinsufficiency. The HRM and wild type mice show similar levels of overall activity, coordination, thermal nociception, startle responses, and anxiety-like behavior. In addition, both genotypes show similar shock threshold, identical cued freezing behavior and comparable spatial learning in Morris water maze tasks. However, a significant reduction in contextual fear conditioned learning was observed in the HRM. Electrophysiological studies in hippocampal CA1 synapses revealed a plethora of differences between genotypes. The HRM exhibits reduced field excitatory postsynaptic potentials in responses to similar synaptic inputs, lowered paired pulse facilitation ratio and impaired long-term depression and tetanus-induced long-term potentiation (LTP). Also, deficits were detected in LTP elicited by theta burst stimulation or by a whole cell pairing protocol. These physiologic differences could not be accounted for by changes in the overall amount of glutamate receptor subunits. In addition, it was determined that network-driven excitatory and inhibitory activities recorded in CA1 pyramidal neurons showed that the HRM had comparable amplitude and frequency of spontaneous excitatory postsynaptic currents, but a marked reduction in spontaneous inhibitory postsynaptic currents. Thus, the HRM exhibits a specific hippocampal dependent learning deficit accompanied with a pronounced impairment of hippocampal plasticity and functional inhibitory innervation. PMID- 16376117 TI - Plasma D-lactate as a potential early marker for colon ischaemia after open aortic reconstruction. AB - BACKGROUND AND AIM: The breakdown of mucosal barrier function due to intestinal hypo-perfusion is the earliest dysfunction of ischaemic colitis. Severe colon ischaemia after aortic reconstruction is associated with mortality rates up to 90%. Therefore, early detection and treatment of patients with extensive ischaemic colitis is of crucial importance. In experimental studies, both D lactate and bacterial endotoxin have been reported as markers of intestinal mucosal barrier impairment. However, evidence of their value in clinical practice is lacking. The aim of this pilot prospective cohort study was to assess the association between ischaemia of the colon (assessed histologically) and plasma levels of D-lactate and endotoxin in patients undergoing open aortic reconstruction. PATIENTS AND METHODS: Twelve consecutive patients underwent surgery between February and April 2003. Six patients underwent emergency surgery and six patients elective aortic surgery. D-Lactate and endotoxin levels were measured in blood samples collected according to a standardised protocol. For histological examination biopsies were obtained by sigmoidoscopy on days 4-6 after surgery, or earlier if indicated clinically. RESULTS: As early as 2 h postoperatively, elevated plasma levels of d-lactate were measured in patients with histologically proven ischaemic colitis. The peak of D-lactate elevation was on postoperative days 1 and 2. Concentration of plasma endotoxin was not significantly different in patients with or without ischaemic colitis. CONCLUSION: Our data suggest that plasma D-lactate levels are a useful marker for early detection of ischaemic colitis secondary to aortic surgery. PMID- 16376118 TI - Cement embolization into the vena cava and pulmonal arteries after vertebroplasty: interdisciplinary management. AB - PURPOSE: To report interdisciplinary management in a case of cement embolization into the inferior vena cava and peripheral pulmonary arteries after percutaneous vertebroplasty. CASE REPORT: A 50-year-old female patient with an osteoporotic compression fracture of the second lumbar vertebra underwent percutaneous vertebroplasty with polymethylmetaacrylate. Thereafter, CT scanning revealed small asymptomatic cement emboli in peripheral pulmonary arteries, along with a hook-shaped cement fragment in the inferior vena cava. Due to the risk that the large cement fragment could migrate to the pulmonary arteries and cause serious complications, they were retrieved from the inferior vena cava by an endovascular technique and extracted through a surgical groin incision. The patient received anticoagulant treatment for 3 months and is free of complaints after 1 year. CONCLUSION: This case shows that this rare complication following vertebroplasty can be successfully managed with an interdisciplinary approach. PMID- 16376119 TI - The distal zone of the deep femoral artery as the inflow site in femorodistal bypass grafting. AB - The distal zone of the deep femoral artery has been used as an inflow site for femorodistal bypass grafts. Eleven such bypass grafts were performed in patients who had critical limb ischemia either to avoid a hostile groin (n=7) or because of an insufficient length of vein (n=4). The posteromedial route was used. Two of the 11 grafts had early failure and a third failed at 3 months: redo surgery restored arterial blood flow to the limb in all three patients. The procedure was safe and is a widely applicable, effective alternative that extends the limits of infrainguinal vein bypass grafting. PMID- 16376120 TI - The retroperitoneum protects prosthetic graft material from intraperitoneal contamination: an experimental study. AB - OBJECTIVES: To evaluate the ability of the retroperitoneum to serve as a barrier, against bacterial contamination, between the peritoneal cavity to the retroperitoneal space. METHODS: Seventy rats had a small piece of knitted Dacron graft placed in the retroperitoneal space and 10(6)-10(9) colony forming unit (cfu) Enterococcus faecalis was injected into the peritoneal cavity. In half the retroperitoneal (RP) group, the retroperitoneum was preserved and in the remainder, the open peritoneal (OP) group, needle holes were created. Grafts were harvested after 1, 4, or 7 days and cultured for E. faecalis. A blood sample was collected from three rats in each group for culture before the graft was harvested. RESULTS: Graft infection did not develop in any rat injected with 10(6) or 10(7) cfu in the RP group, while seven out of the 10 graft cultures of the OP group grew E. faecalis (P = 0.003). In rats injected with 10(8) or 10(9) cfu, five out of the 10 graft cultures in the RP group and eight out of 10 in the OP group grew E. faecalis. All blood cultures were negative when the injected bacterial count was 10(7) cfu or less. One out of the three blood cultures was positive at 10(8) cfu, and all were positive at 10(9) cfu. CONCLUSIONS: These results suggest that an intact retroperitroneum acts as a protective barrier against intraperitoneal bacterial contamination, particularly when blood cultures are negative. PMID- 16376121 TI - Duplex scan evaluation of carotid stenosis. Which consensus is necessary? PMID- 16376122 TI - Spontaneous resolution of carotid stenosis--a case for routine preoperative duplex ultrasound. AB - Spontaneous resolution of carotid stenosis has not been previously reported in the context of preoperative duplex ultrasound scanning, although it has been described as a recognizable phenomenon in the past. We report a case in whom significant carotid stenosis was noted at the time of listing for surgery on both duplex ultrasound and MRA. On preoperative imaging there was resolution of the lesion and surgery was avoided. This case emphasises that spontaneous resolution of carotid stenosis can occur and that preoperative duplex is useful as a prelude to surgery and can prevent unwarranted intervention. PMID- 16376123 TI - The small abdominal aortic aneurysm. PMID- 16376124 TI - Complicated acute type B dissections--an 8-years experience of endovascular stent graft repair in a single centre. AB - OBJECTIVE: To analyze the experience of a single centre using stent-grafts for treatment of complicated acute aortic type B-dissections (EVR-ABD). DESIGN: Retrospective analysis of prospectively collected data from patients undergoing EVR-ABD between January 1997 and December 2004. METHODS: EVR-ABD was performed in 31 patients (20 males, median age 74 years (IQR: 64-79)). Indications for treatment were aortic rupture (22 patients), intractable pain and hypertension (six patients), acute bowel ischemia (two patients) and transient paraplegia, lower limb and renal ischemia in one patient. Initially home-made devices (five patients) and subsequently commercially available thoracic stent-grafts were used. RESULTS: Five patients (16%) died within 30 days of EVR-ABD. Postoperative complications occurred in 15 (48%) patients, including one paraplegia converted to paraparesis after cerebrospinal fluid drainage, five strokes, three lower limb ischemia, three myocardial infarction, two pneumonia and one colitis). Re interventions were required in nine patients (29%). Six more deaths occurred during a median follow-up of 22 (IQR: 16-34) months, two related to the stent graft and four due to cardiac disease. CONCLUSIONS: Stent-graft repair of complicated acute type B dissections seems to provide acceptable results and, therefore, it may be considered a valuable alternative to open surgery. PMID- 16376125 TI - Emerging research on the treatment and etiology of secondary or comorbid insomnia. AB - Secondary insomnia is defined as difficulty initiating or maintaining sleep that occurs as a consequence of another primary medical or psychiatric disorder. This distinction from primary insomnia has been thought to be important from a diagnostic standpoint so that treatment for secondary insomnia can appropriately target the primary disorder. By convention, treating insomnia directly has only been recommended with primary insomnia. However, a number of recent studies of cognitive-behavioral treatment for a wide range of secondary insomnias have demonstrated that this approach is an effective treatment in this population. These results are not predicted by the traditional conceptualization of secondary insomnia. Results of epidemiological research also argue against viewing insomnia that is comorbid with another disorder as being caused by that disorder. A revised model is needed to understand insomnia that is comorbid with medical and/or psychiatric illness, in addition to re-evaluating standard treatment protocols for comorbid insomnia. PMID- 16376126 TI - Digestibility of a complete ration in horses fed once or three times a day and correlation with key blood parameters. AB - To determine the influence of feeding frequency on apparent digestibility and blood metabolites in horses, four geldings were fed a complete ration either once (at 08:00 h) or three times a day (at 08:00, 13:00 and 18:00 h). Horses were provided with a mixture of cereals and chopped alfalfa hay at maintenance level for energy supply, 344 kJ/kg BW(0.75) on a daily basis. After three weeks' adaptation, total amounts of faeces and urine were collected for five days, using collection harnesses. Serial blood samples were taken at -30, -15, 0, 5, 15, 30, 45, 60, 90, 120, 180, 240, 300, 360, 480 and 600 min after feeding and analysed for glucose, L-lactate, triglycerides (TG), non-esterified free fatty acids (NEFA) and triiodothyronine (T(3)). Apparent digestibility coefficients of dry matter (DM), crude protein (CP), ether extract (EE), crude fibre (CF), ash and nitrogen-free extract (NFE) were not significantly different (P>0.05) between the two feeding frequencies. Basal plasma glucose, plasma glucose curves and other plasma blood metabolites were not affected by meal frequency. The horses fed the whole ration in one morning feeding did not consume the entire meal at once, but spared feed for ingestion throughout the day, which may explain the lack of effect. Feeding frequency did not affect plasma glucose response (area under curve :AUC) (P=0.705), but AUC (glucose) was negatively correlated with CP (R(2)=0.76; P=0.005) and CF digestibility (R(2)=0.61; P=0.022). Further research is needed to clarify whether different endocrine responses or differences in passage rate can explain these correlations. PMID- 16376127 TI - Dental and skeletal characterization of the BR-1 minipig. AB - This study was designed to characterize the development of oral structures in the BR-1 minipig and to compare these data with literature reports on domestic swine and other minipig lineages. Twelve male minipigs were anesthetized and given physical and radiographic examinations. The presence and chronology of tooth eruption from 3 to 15 months of age were assessed, and the radiographic results were interpreted by two certified researchers, who obtained a high index of agreement (kappa>0.81). The eruption chronology was similar to that reported for domestic swine and other minipig lineages. The results indicate that it is possible to use the BR-1 minipig in dental research. PMID- 16376128 TI - Stage-dependent analgesia of electro-acupuncture in a mouse model of cutaneous cancer pain. AB - Acupuncture is one of the most effective alternative medical treatments in pain management with the advantages of simple application, low cost and minimal side effects. However its scientific evidence and laws of action are not very clear in cancer pain relieving. The aim of this study was to examine the immediate and therapeutic anti-hyperalgesic effect of electro-acupuncture (EA) on a mouse model of cutaneous cancer pain. B16-BL6 melanoma cells were inoculated into the plantar region of unilateral hind paw and the thermal hyperalgesia was measured by using radiant heat test and hot plate test. C57BL/6 mice showed moderate and marked hyperalgesia during days 8-12 and from day 14 after the orthotopic inoculation of B16-BL6 melanoma cells into the hind paw. Single EA on day 8 after inoculation showed significant analgesic effect immediately after the treatment, the analgesic effect reached its maximum within 15-30min and declined to its minimum at 50min after EA treatment. Single EA treatment on day 20 showed no significant analgesic effect; Repeated EA treatments (started from day 8, once every other day) showed therapeutic analgesic effect, while it showed no therapeutic effect when started from day 16, a relatively late stage of this cancer pain model. The results demonstrated that EA had anti-hyperalgesic effect on early stage of cutaneous cancer pain but not on late stage. These results indicated a tight correlation of EA anti-hyperalgesic effects with the time window of cancer pain. PMID- 16376129 TI - Comparisons of innate immune activity of all known living crocodylian species. AB - Serum samples from all twenty-three known living members of the Crocodylia were tested for antibacterial activity against eight bacterial species. These data were used to generate an immune profile for each crocodylian species. Statistical analyses revealed that the three living lineages of crocodylians, Alligatoroidea, Crocodyloidea, and Gavialoidea, were distinguishable by their immunological activities. For instance, species within the Alligatoroidea and Crocodyloidea exhibited remarkable immune activity similarities to others in their own lineages. Comparisons of the members of the different lineages, however, revealed substantial differences in immune profiles. Furthermore, species that are in the same genus were shown to exhibit more immune similarities to each other than to members of other genera within the same family. Finally, our immunological analyses reveal that Tomistoma schlegelii aligns more closely with the Gavialoidea than the Crocodyloidea. PMID- 16376130 TI - Colored Petri net modeling and simulation of signal transduction pathways. AB - Presented herein is a methodology for quantitatively analyzing the complex signaling network by resorting to colored Petri nets (CPN). The mathematical as well as Petri net models for two basic reaction types were established, followed by the extension to a large signal transduction system stimulated by epidermal growth factor (EGF) in an application study. The CPN models based on the Petri net representation and the conservation and kinetic equations were used to examine the dynamic behavior of the EGF signaling pathway. The usefulness of Petri nets is demonstrated for the quantitative analysis of the signal transduction pathway. Moreover, the trade-offs between modeling capability and simulation efficiency of this pathway are explored, suggesting that the Petri net model can be invaluable in the initial stage of building a dynamic model. PMID- 16376131 TI - PMM2 intronic branch-site mutations in CDG-Ia. AB - Congenital Disorders of Glycosylation (CDG, OMIM#212065)-Ia is an autosomal recessive disorder, characterized by central nervous system dysfunction and multiorgan failure associated with mutations in the PMM2 gene. We report two patients who are compound heterozygotes with respect to two new intronic mutations that affect a highly conserved adenosine in a consensus branch-site sequence. The mutations, one in intron 7: c.340 -23A > G (IVS7 -23A > G) and the other in intron 2: c.179 -25A > G (IVS2 -25A > G), are associated with the c.422G > A (R141H) and c.193 G > T (D65Y) mutations, respectively. The c.179 -25A > G and the c.340 -23A > G changes cause exon 3 and exon 8 to be lost at the RNA level, respectively. This kind of mutation can cause a problem in molecular diagnosis of CDG-Ia if intronic primers are not correctly chosen, and if molecular diagnosis is not performed at both the DNA and mRNA levels. PMID- 16376132 TI - Functional analysis of acyl-CoA dehydrogenase catalytic residue mutants using surface plasmon resonance and circular dichroism. AB - The acyl-CoA dehydrogenases (ACDs) are a family of flavoenzymes involved in the metabolism of fatty acids and branched-chain amino acids. The ACDs share a similar structure and a common dehydrogenation mechanism in which a catalytic glutamate extracts a proton from an acyl-CoA substrate. The resulting charge transfer complex subsequently passes electrons to electron-transferring flavoprotein (ETF). We previously generated catalytic residue mutants of human short-chain acyl-CoA dehydrogenase (SCAD) and isovaleryl-CoA dehydrogenase (IVD) that were difficult to characterize by traditional methods. In the present study, we developed a novel surface plasmon resonance-based assay to measure substrate binding to these mutants. Replacement of the catalytic glutamate in either SCAD or IVD with glycine resulted in a several-fold reduction in affinity for substrate. Circular dichroism studies substantiated our earlier findings that both SCAD E368G and IVD E254G are unable to form a charge-transfer complex with substrate/product. The CD spectra of IVD E254G also indicated a perturbation of the flavin environment, a finding supported by molecular modeling that predicted a shift in the conformation of a conserved tryptophan that lies in close proximity to the flavin. Lastly, competitive inhibition studies using the ETF fluorescence reduction assay suggested that SCAD E368G and IVD E254G do not effectively compete with the wild-type enzymes for the physiological electron acceptor ETF. PMID- 16376133 TI - Interactions of C-reactive protein with low-density lipoproteins: implications for an active role of modified C-reactive protein in atherosclerosis. AB - The interaction of C-reactive protein with low-density lipoprotein is considered to be one of the key properties that link C-reactive protein with atherosclerosis. However the data obtained to date are controversial, and hence make it difficult to conclude actual physiological or pathological impact of such interaction. The incompatible findings could be ascribed to the different structural state of C-reactive protein and/or low-density lipoprotein. We investigated in detail the interaction of various C-reactive protein isoforms with native and modified low-density lipoprotein. Our data showed "C-reactive protein" could indeed interact with each of native low-density lipoprotein, oxidized or enzymatically modified low-density lipoprotein, but that interaction occurs primarily when C-reactive protein is conformed in a modified form and not pentameric structure. Low level of modified C-reactive protein "contaminant" could confer C-reactive protein obvious low-density lipoprotein binding capacity. Interaction of modified C-reactive protein and low-density lipoprotein was mediated synergistically by both electrostatic association with ApoB and hydrophobic insertion into lipid layer. When complexed with modified C-reactive protein, macrophage binding/uptake of native and oxidized low-density lipoprotein was either increased 150% or decreased 35%, respectively. Thus the interaction of modified C-reactive protein with low-density lipoprotein may contribute to the regulation of low-density lipoprotein metabolism and foam cell formation in arterial wall. These results highlight an active role of modified C-reactive protein in atherosclerotic process. PMID- 16376134 TI - Recent advances of protein microarrays. AB - Technological innovations and novel applications have greatly advanced the field of protein microarrays. Over the past two years, different types of protein microarrays have been used for serum profiling, protein abundance determinations, and identification of proteins that bind DNA or small compounds. However, considerable development is still required to ensure common quality standards and to establish large content repertoires. Here, we summarize applications available to date and discuss recent technological achievements and efforts on standardization. PMID- 16376135 TI - Swallowing after free-flap reconstruction in patients with oral and pharyngeal cancer. AB - Swallowing and intraoral sensation outcome were investigated prospectively after microvascular free-flap reconstruction. Forty-one patients with a large oral or oropharyngeal carcinoma underwent free-flap surgery usually combined with radiotherapy. The patients completed modified barium swallow, self-rating of swallowing, and 2-point moving discrimination preoperatively and at four time points during the 12-month follow-up period, and a plain chest X-ray one year after operation. Swallowing was impaired with respect to an objective and subjective measure after therapy. Rates for nonsilent and silent aspiration increased during the follow-up. Intraoral sensation deteriorated. Swallowing outcome was not related to sensation. One year after surgery, 86% of the patients ate regular masticated or soft food. Microvascular transfers offer a reasonable option for oral reconstruction. This study does not support the need for sensate flaps. Swallowing problems should be routinely sought and patients rehabilitated during a sufficiently long follow-up with videofluorography regardless of the patient's perception of swallowing. PMID- 16376136 TI - Amphiregulin as a tumor promoter for oral squamous cell carcinoma: involvement of cyclooxygenase 2. AB - Amphiregulin (AR), an epidermal growth factor (EGF)-like molecule, is a mitogen for keratinocytes. Squamous cell carcinoma (SCC) is a tumor derived from keratinoctyes. Expression of AR mRNA positively correlated with the clinical progression of 39 oral SCC. Oral SCC line, KB, was used as a model to study if increased expression of AR altered the biological behaviors of SCC cells. Exogenous AR dose-dependently enhanced the proliferation of KB cells expressing EGF receptor 1 (EGFR-1), a major receptor for AR, but little AR. Neutralizing anti-AR antibody significantly reversed the stimulatory effect of exogenous AR on KB cell proliferation. Ectopically expressed AR in stable clones manifested higher abilities to proliferate, migrate and invade Matrigel than vector control. Cyclooxygenase 2 (COX-2), but not metalloprotease 9 (MMP-9) mRNA, was increased in all the AR-expressing stable clones. In summary, AR behaves as a tumor promoter for oral SCC cells partly via increased expression of COX-2. PMID- 16376137 TI - Continuity of care and early diagnosis of head and neck cancer. AB - This study investigated whether receiving continuous regular care with the same primary care provider had an independent association with stage at diagnosis for head and neck cancers (HNCA) in a Medicare population. The study database (N = 10,662) linked files from the Surveillance, Epidemiology, and End Results Program 1991-2000 for patients with cancers of the oral cavity, pharynx, and larynx with their files from the Center for Medicare and Medicaid Services Program. Continuity of care displayed a statistically significant independent association and dose-response pattern with stage at diagnosis when the provider was an internist but not a general or family physician. An independent statistically significant association between continuity of care with an internist and stage at diagnosis was found for oral cavity tumors (adjusted odds ratio [AOR] = 0.85; 95%CI = 0.73-0.99), but not pharyngeal tumors (AOR = 1.02; 95%CI = 0.74-1.41). Laryngeal tumors were borderline statistically significant at p-value = 0.07 (AOR = 0.84; 95%CI = 0.69-1.02). Regular continuous care from the same primary care provider displayed an independent association with early diagnosis of HNCA when the anatomic site was the oral cavity and the provider was an internist, but not when the provider was a general or family physician. Primary care physicians including general/family physicians may represent a relevant group for encouraging increased opportunistic HNCA screening efforts among their patients. PMID- 16376138 TI - Epithelial expression of SHH signaling pathway in odontogenic tumors. AB - Sonic hedgehog (SHH) signaling pathway plays a critical role in tooth development. Recent studies indicate that SHH signaling pathway activation occurs both in the odontogenic cyst and ameloblastoma. However, the association of SHH pathway with other subtypes of odontogenic tumor is not well documented. The objective of this paper is to investigate the protein distribution of SHH and its receptor PTC, SMO and transcription factor GLI1 in various odontogenic tumors. Odontogenic tumor tissues including 34 epithelial derived, 24 epithelial mesenchymal derived, and 26 mesenchymal derived were examined by immunohistochemistry for SHH, PTC, SMO and GLI1. Immunoreactivity for SHH, PTC, SMO and GLI1 was detected in both epithelial derived odontogenic tumors and epithelial-mesenchymal derived odontogenic tumors with or without dental hard tissue formation. Mesenchymal derived odontogenic tumors showed no positive staining except for the focal epithelial cells in island or cord forms within the central portion of the tumor. The protein expression of SHH signaling pathway in malignant odontogenic tumors was no stronger than that in benign tumors. Each of the genes in the pathway was expressed in similar patterns in the same tumor subtype. SHH, PTC, SMO and GLI1 were detected more in the cytoplasm of the epithelial cells than in stromal cells. Immunoreactivity for GLI1 was also detected in the base membrane of the tumor cells. The findings suggest SHH, PTC, SMO and GLI1 protein are predominantly located in epithelial components in various odontogenic tumors and might participate in the proliferation of epithelial components of odontogenic tumors. PMID- 16376139 TI - Historical aspects for the control of soil-transmitted helminthiases. AB - Japan has been one of the very few countries that operated the nationwide control programme against soil-transmitted helminths (STH) infections and gained a great success. On the basis of researches and operational studies, periodical mass examination using cellophane thick smear (Kato technique) and selective mass treatment targeted at school children were employed as the most useful control measure. In later years, Japan's experience and the strategy for STH control programme were transferred to Asian countries through Asian Parasite Control Organization (APCO) and also to countries of Asia, Africa and Latin-America through seminar at a modified form of the integration programme (IP) of STH control with family planning and nutritional improvement. The parasitologists group conducted studies in all aspects to seek effective STH control measures for respective countries and obtained many successful results. PMID- 16376140 TI - Taeniasis and cysticercosis in Bali and North Sumatra, Indonesia. AB - It has been reported that three human Taenia species are distributed in Indonesia: Taenia solium, Taenia asiatica and Taenia saginata. T. asiatica is well known in North Sumatra, especially on Samosir island in Lake Toba. T. solium and T. saginata are known from Bali. T. solium is most serious public health issue in Papua (former Irian Jaya). In this report, we briefly review the present situation of these three human Taenia species mainly in Bali and North Sumatra. For community based epidemiological survey, we have adopted and applied questionnaire, microscopic observation of eggs, coproantigen tests, coproDNA tests, mitochondrial DNA analysis and morphology for isolated specimens, serology for cysticercosis and taeniasis. For detection of taeniasis cases of both T. saginata and T. asiatica, questionnaire by expert doctors is perfect to detect worm carriers. Corpoantigen test developed for detection of taeniasis of T. solium is also highly reliable to detect taeniasis carriers in the community. Epidemiological data from these two islands 2002-2005 are reviewed with backgrounds of historical culture, religions and customs. PMID- 16376141 TI - Transcranial magnetic stimulation--a sandwich coil design for a better sham. AB - OBJECTIVE: To improve the quality of TMS studies by developing a new sham condition. METHODS: We describe a novel and easily arranged TMS set-up of two standard TMS coils and a magnetic shield, stacked like a sandwich. In a first step we compare the magnetic field in the sham and verum conditions. In a second step we ask six subjects to rate the stimulation intensity. RESULTS: The magnetic field in the sham mode is reduced to about one eighth of that during verum stimulation. The attenuation of the magnetic field is not limited to the actual stimulation site but also effective at neighbouring brain areas, avoiding direct and indirect stimulation via connected neural pathways. This also minimizes stimulation of the skin, but as a consequence allows subjects to distinguish between verum and sham conditions when these are contrasted directly. The position of the coil system and the acoustic sensations are indistinguishable between sham and verum condition. Subjects are not able to discriminate TMS position and condition by external cues. CONCLUSIONS: The proposed TMS setup is simple and allows verum and sham TMS without interaction of the researcher. If used with the magnetic shield, the magnetic field in the brain is attenuated most. SIGNIFICANCE: With the sandwich TMS coil system it is possible to improve the quality of TMS studies. PMID- 16376142 TI - Location-specific and task-dependent modulation of cutaneous reflexes in intrinsic human hand muscles. AB - OBJECTIVE: The current study was designed to determine location-specificity in long latency cutaneous reflexes in intrinsic human hand muscles while performing a simple abduction and a manual task. METHODS: Subjects comprised of 13 neurologically intact healthy volunteers. Cutaneous reflexes following non noxious electrical stimulation to the digits of the hand (digit 1, D1; digit 2, D2; and digit 5, D5) were elicited while the subjects performed isolated isometric contraction of the abductor pollicis brevis (APB), first dorsal interosseous (FDI) and abductor digiti minimi muscles (ADM). The cutaneous reflexes were also elicited while the subjects performed a pincer grip with D1 and D2 while slightly lifting the hand from the supporting surface by abduction of D5 (manual task). RESULTS: While performing isolated tonic voluntary contraction of the APB, FDI and ADM, the magnitude of E2 (peak latency approximately 60-90 ms) was larger when stimulation was delivered to the homotopic digit (e.g. APB response following D1 stimulation) than to the heterotopic nearby (e.g. APB response following D2 stimulation) or heterotopic distant digit (e.g. APB response following D5 stimulation). I2 ( approximately 90 120 ms) and E3 ( approximately 120-180 ms) were significantly larger following D5 stimulation than D1 or D2 stimulation in all muscles tested. The size of each component in the ADM following D1 and D2 stimulation did not increase even when the contraction level of the ADM increased. However, while performing the manual task, the E2 response in the ADM following both D1 and D2 stimulation was significantly increased as compared to that recorded during isolated D5 abduction. CONCLUSIONS: Long latency cutaneous reflexes following non-noxious electrical stimulation are organized in a highly location-specific as well as task-dependent manner. SIGNIFICANCE: Our findings provide further insight into the nature and functional significance of long latency cutaneous reflexes in human intrinsic hand muscles. PMID- 16376143 TI - Sources and effects of electrode impedance during deep brain stimulation. AB - OBJECTIVE: Clinical impedance measurements for deep brain stimulation (DBS) electrodes in human patients are normally in the range 500-1500 Omega. DBS devices utilize voltage-controlled stimulation; therefore, the current delivered to the tissue is inversely proportional to the impedance. The goals of this study were to evaluate the effects of various electrical properties of the tissue medium and electrode-tissue interface on the impedance and to determine the impact of clinically relevant impedance variability on the volume of tissue activated (VTA) during DBS. METHODS: Axisymmetric finite-element models (FEM) of the DBS system were constructed with explicit representation of encapsulation layers around the electrode and implanted pulse generator. Impedance was calculated by dividing the stimulation voltage by the integrated current density along the active electrode contact. The models utilized a Fourier FEM solver that accounted for the capacitive components of the electrode-tissue interface during voltage-controlled stimulation. The resulting time- and space-dependent voltage waveforms generated in the tissue medium were superimposed onto cable model axons to calculate the VTA. RESULTS: The primary determinants of electrode impedance were the thickness and conductivity of the encapsulation layer around the electrode contact and the conductivity of the bulk tissue medium. The difference in the VTA between our low (790 Omega) and high (1244 Omega) impedance models with typical DBS settings (-3 V, 90 mus, 130 Hz pulse train) was 121 mm3, representing a 52% volume reduction. CONCLUSIONS: Electrode impedance has a substantial effect on the VTA and accurate representation of electrode impedance should be an explicit component of computational models of voltage-controlled DBS. SIGNIFICANCE: Impedance is often used to identify broken leads (for values > 2000 Omega) or short circuits in the hardware (for values < 50 Omega); however, clinical impedance values also represent an important parameter in defining the spread of stimulation during DBS. PMID- 16376144 TI - Interaction between the flash evoked SSVEPs and the spontaneous EEG activity in children and adults. AB - OBJECTIVE: To evaluate the interaction between the steady-state visual evoked potentials (SSVEPs) recorded during the intermittent photic stimulation (IPS) and the spontaneous EEG activities both in children and adults. METHODS: EEG was recorded during the rest and under 5, 7.5, 10 and 12.5 Hz IPS in 41 children between 3 and 16 years and 10 adults. We distinguished between the spontaneous resting EEG spectra, SSVEPs (1st harmonic) and undriven (ongoing) EEG spectra recorded during the IPS. RESULTS: We show that IPS influences spontaneous EEG activity by specifically suppressing or desynchronizing individual posterior dominant resting EEG frequencies (DF) in both children and adults. Further, this highly significant and consistent suppressing effect positively correlates with the SSVEPs amplitude. CONCLUSIONS: Our data suggest that the desynchronization of the spontaneous EEG activity under IPS and the SSVEPs are related to each other. SIGNIFICANCE: These relationships could be interesting to study in pathological conditions where the neural synchronization and the responses to IPS have been shown to be affected, such as epilepsy and schizophrenia. PMID- 16376145 TI - Is disinfection of mechanical ventilation tubing needed at home? AB - INTRODUCTION: Home mechanical ventilation is used to treat chronic alveolar hypoventilation. Maintenance protocols for home ventilation circuits (HVC) remain empirical and unproven. We have investigated (1) the cleanliness and sterility of the HVC used by home ventilated patients and (2) the efficiency of tubing cleaning and decontamination protocols recommended to them and used for 12 months or more. METHOD: HVC cleanliness was assessed in 39 severe restrictive ventilated patients (16 (T) tracheostomy vs. 23 (N) noninvasive) and in 7 new valves as control. In the first experiment (Exp1), a visual and bacteriologic inspection of the expiratory valve (Eva) was conducted during a consultation in our centre. Eva visual cleanliness was assessed on a 10-point scale and Eva bacteriologic contamination analysis was performed on a dry smear. In the second experiment (Exp2), these analyses were repeated after a cleaning sequence chosen at random, either chemical (ammonium-chlorhexidine complex) (A) or mechanical by dishwasher (B). RESULTS: In Exp1, 69% of Eva were dirty. Dirtiness was worse in (T) than in (N) (5.3 vs. 2; p<0.001). There was a significant positive correlation between visual cleanliness and bacteriologic contamination (r=0.56; p<0.001). Eva in group (T) were more contaminated than in group (N) (p<0.001). Eva contamination rates reached 22% in group (N) but without the presence of any potentially pathogenic organisms (PPO) and 81% in group (T) where 19% were PPO. In Exp2, EVA visual cleanliness was better after dishwasher cleaning (B) compared to chemical (A) (0.16 vs. 1.05; p<0.001) with similar bacteriological decontamination. CONCLUSION: HVC from noninvasive ventilated patients are dirty but not contaminated by PPO. We recommend washing them in a dishwasher or with detergent and hot water without specific disinfection. PPO contaminated 1/5 of invasive HVC, for which we recommend dishwasher cleaning. Decontamination is only indicated when tubing is visually very dirty or/and when tracheostomized patients are particularly sensitive to respiratory tract infections. The expiratory valve must be carefully washed specifically, with care that its balloon is not placed under water. PMID- 16376146 TI - Information needs and experiences: an audit of UK cancer patients. AB - As part of a multi-centred UK study evaluating multidisciplinary team communication, the information needs, decision making preferences and information experiences of 394 cancer patients were audited. A majority of patients (342/394, 87%) wanted all possible information, both good and bad news. Assuming that all clinicians had equal skill, the majority of patients (350/394, 89%) expressed no preference for the sex of their doctor. The largest proportion of patients (153/394, 39%) wanted to share responsibility for decision making, preference was significantly influenced by age (chi2=17.42, df=4 P=0.002) with older patients more likely to prefer the doctor to make the decisions. A majority of patients reported receiving information regarding their initial tests (313/314, 100%), diagnosis (382/382, 100%), surgery (374/375, 100%) and prognosis (308/355, 87%), fewer recalled discussions concerning clinical trials (119/280, 43%), family history (90/320, 28%) or psychosocial issues, notably sexual well-being (116/314, 37%). Cancer patients want to be fully informed and share decision making responsibility, but do not report receiving sufficient information in all areas. Multidisciplinary cancer teams need to ensure that where appropriate, someone provides patients with information about clinical trials, familial risk and psychosocial issues. Regular audits highlight gaps and omissions in the information given to patients. PMID- 16376147 TI - The mechanisms of action of gabapentin and pregabalin. AB - Gabapentin and pregabalin are structurally related compounds with recognized efficacy in the treatment of both epilepsy and neuropathic pain. The pharmacological mechanisms by which these agents exert their clinical effects have, until recently, remained unclear. The interaction of gabapentin and pregabalin with conventional antiepileptic and analgesic drug targets is likely to be modest, at best, and has been largely dismissed in favour of a selective inhibitory effect on voltage-gated calcium channels containing the alpha2delta-1 subunit. This mechanism is consistently observed in both rodent- and human-based experimental paradigms and may be sufficiently robust to account for much of the clinical activity of these compounds. PMID- 16376148 TI - Glutamate-based therapeutic approaches: inhibitors of glycine transport. AB - A growing body of evidence suggests that activation of the glutamatergic system, particularly N-methyl-D-aspartate (NMDA) receptor function, may be a viable approach to the treatment of schizophrenia, and potentially other cognitive disorders. The excitotoxicity associated with direct NMDA receptor agonists limits their therapeutic potential, and the glycine modulatory site of the NMDA receptor has received growing interest as a therapeutic target. One approach to enhance NMDA receptor function is to increase the availability of the necessary co-agonist glycine at this modulatory site through inhibition of glycine reuptake from the synapse via glycine transporter-1 (GlyT1). Both preclinical and clinical evidence provide support for this approach, as do recent findings demonstrating the regulation of dopaminergic neurotransmission by GlyT1 inhibition. As a result, several groups have focused on the development of novel GlyT1 inhibitors. In addition, recent electrophysiological findings and data from transgenic mouse models suggest that GlyT1 might also play a role in terminating the actions of glycine at strychnine-sensitive glycine receptors, and therefore GlyT1 antagonists also have potential for the treatment of conditions where activation of inhibitory pathways in the central nervous system might be beneficial. PMID- 16376149 TI - Glutamate-based therapeutic approaches: NR2B receptor antagonists. AB - Over the past decade, there have been major advances in our understanding of the role of glutamate and N-methyl-d-aspartate (NMDA) receptors in several disorders of the central nervous system, including stroke, Parkinson's disease, Huntington's disease and chronic/neuropathic pain. In particular, NR2B subunit containing NMDA receptors have been the focus of intense study from both a physiological and a pharmacological perspective, with several pharmaceutical companies developing NR2B subtype-selective antagonists for several glutamate mediated diseases. Recent studies have shown the importance of NR2B subunits for NMDA receptor localization and endocytosis, and have suggested a role for NR2B containing NMDA receptors in the underlying pathophysiology of neurodegenerative disorders such as Alzheimer's and Huntington's diseases. Anatomical, biochemical and pharmacological studies over the past five years have greatly added to our understanding of the role of NR2B subunit-containing NMDA receptors in chronic and neuropathic pain states, and have shown that NR2B-mediated analgesic effects might be supra- rather than intra-spinally mediated, and that phosphorylation of the NR2B subunit could be responsible for the initiation and maintenance of the central sensitization seen in neuropathic pain states. These data will hopefully provide the impetus for development of novel compounds that use multiple approaches to modulate the activity of NR2B subunit-containing NMDA receptors, thus bringing to fruition the promise of therapeutic efficacy utilizing this approach. PMID- 16376151 TI - ICAT-based comparative proteomic analysis of non-replicating persistent Mycobacterium tuberculosis. AB - The non-replicating persistence (NRP) phenotype of Mycobacterium tuberculosis (NRP-TB) is assumed to be responsible for the maintenance of latent infection and the requirement of a long treatment duration for active tuberculosis. Isotope coded affinity tag-based proteomic analysis was used for the determination of the relative expression of large numbers of M. tuberculosis proteins during oxygen self-depletion under controlled conditions in a multi-chambered fermentor. Expression of the alpha-crystallin homolog protein, acr, was monitored and quantified to confirm entry into NRP. Relative expression of 586 and 628 proteins was determined in log phase vs. early stage NRP (NRP-1) and log phase vs. later stage NRP (NRP-2), respectively. Relative to expression in log phase and using an abundance ratio of +/-2.0 as a cutoff, 6.5% and 20.4% of proteins were found to be upregulated in NRP-1 and NRP-2, respectively while 20.3% and 13.4% were downregulated, respectively. Functional profiling revealed that 42.1%/39.8% of upregulated proteins and 41.2%/45.2% of downregulated proteins in NRP-1/NRP-2, respectively, were involved in small molecule metabolism. Among those proteins the highest proportions of 37.5% in NRP-1 were involved with degradation and of 45.1% in NRP-2 with energy metabolism. These results suggest distinct protein expression profiles in NRP-1 and NRP-2. PMID- 16376150 TI - GABA-based therapeutic approaches: GABAA receptor subtype functions. AB - It is increasingly being appreciated that GABAA receptor subtypes, through their specific regional, cellular and subcellular localization, are linked to distinct neuronal circuits and consequently serve distinct functions. GABAA receptor subtype-selective drugs are therefore expected to provide novel pharmacological profiles. Receptors containing the alpha1 subunit mediate sedation and serve as targets for sedative hypnotics. Agonists selective for alpha2- and/or alpha3 containing GABAA receptors have been shown to provide anxiolysis without sedation in preclinical models, whereas inverse agonists selective for alpha5-containing GABAA receptors provide memory enhancement. Agonists selective for alpha3 containing GABAA receptors might be suitable for the treatment of deficits in sensorimotor processing in psychiatric disorders. Thus, a new pharmacology based on GABAA receptor subtype-specific actions is emerging. PMID- 16376152 TI - Platelet storage lesion: an update on the impact of various leukoreduction processes on the biological response modifiers. AB - Currently, platelet concentrates are produced either from pooled buffy which are leukoreduced during processing, by various types of WBC removal filters and several apheresis technologies, which are leukocyte-reduced during collection with or without filtration. It is therefore important to define the impact of various leukocyte-removal processes on the acceleration of platelet storage of lesion and cellular apoptosis/necrosis. This overview briefly highlights the effects of exposure to artificial surfaces, during apheresis leukoreduction processes and platelet storage bags on the development of the platelet storage lesion that may contribute to transfusion reactions. The results obtained from three plateletpheresis technologies are compared with data from a "like with like" study on buffy-coat-derived platelet concentrates, using three types of platelet filter/pack assemblies. Emphasis is placed on the combined preparative methods and storage bags on generation/removal of: kallikrein/kinin; activated complement, leukocytes and platelet-derived cytokines and the development of cellular injuries, measured by the release of Annexin V. There was no systematic evidence of significant cellular fragmentation caused by filtration, but the combined preparative methods and storage bags appears to have some impact on the rate of release of soluble HLA. Moreover, large variability was observed between and within groups, in terms of various laboratory markers of biocompatibility and major biological response modifiers, indicating that much still remains to be done on various aspects of quality improvement to fully abrogate platelet concentrates associated transfusion reactions. PMID- 16376153 TI - Diet quality in 116 Norwegian men and women with coronary heart disease. AB - BACKGROUND AND AIMS: Clinical intervention studies have provided strong evidence that a healthy diet can prevent coronary heart disease. The aims of this study were to describe the diet quality in Norwegian subjects from Sogn and Fjordane County with coronary heart disease, and to investigate if the diet quality was associated with demographic and lifestyle variables. METHODS: We used a self administered questionnaire to assess information from 116 participants. A Healthy Diet Score (HDS) indicated the diet quality. Multiple Linear regression analyses were used to investigate associations between relevant variables and the HDS. RESULTS: We found that 26.7% of the participants had a poor HDS, 64.6% had a fair HDS and 8.7% had a good HDS. We found that the widespread suboptimal diet quality (poor or fair HDS) was partially related to low consumption of nonhydrogenated mayonnaise products, vegetables, and fruit/berries/juice. Multiple linear regression analysis showed that women had better HDS than men, and that subjects with high education had better HDS than subjects with low education (P < 0.05). CONCLUSION: We found that 91.3% of the participants had a suboptimal diet quality. The consumption of nonhydrogenated mayonnaise products, vegetables and fruit/berries/juice were poor and a principal goal should be to increase the consumption of these food items. Health professionals should keep in mind that gender and educational level may be associated with dietary habits. An important clinical implication of this study is that more attention should be paid to dietary counselling. PMID- 16376154 TI - IRIS and SLE. PMID- 16376155 TI - Hot and hobbling with hives: Schnitzler syndrome. PMID- 16376156 TI - The end of the beginning: oncolytic virotherapy achieves clinical proof-of concept. PMID- 16376157 TI - Vagus nerve stimulation use and effect in epilepsy: what have we learned? AB - Vagus nerve stimulation (VNS) for epilepsy has been available in the United States for 8 years. Pivotal randomized, blinded clinical trials leading to FDA approval in patients age 12 and older with refractory partial seizures have not been performed for other age groups or epilepsy syndromes. This practical review takes stock of the current information about VNS use and efficacy in various types of epilepsy. We review the evidence for commonly used stimulation parameters, end of battery life, predictors of response including duration of epilepsy, seizure type/epilepsy syndrome, bihemispheric seizures, age at implant, and prior cranial surgery. We review adverse events and VNS effects on respiratory patterns, cardiac function, and mood and behavior. With the recent U.S. approval of VNS for treatment-resistant depression, we anticipate that lessons learned from treating patients with epilepsy will be useful to physicians using VNS to treat patients with depression and possibly other conditions. PMID- 16376158 TI - Carbonic anhydrase inhibitors and hypoxic pulmonary vasoconstriction. AB - Acetazolamide and other related carbonic anhydrase (CA) inhibitors have had a long history of effectiveness in prevention and treatment of acute mountain sickness (AMS) and remain the standard of care for this indication. Despite many decades of CA inhibitor use for AMS, the possibility has never been seriously entertained that these drugs might also afford protection against high altitude pulmonary edema (HAPE). In this paper, I will present our evidence and supporting data of others, that acetazolamide has inhibitory effects on the hypoxic response of the pulmonary circulation that may be useful in HAPE. Data from pulmonary artery smooth muscle cells, isolated perfused lungs, and live unanethetized animals all point to a potent reduction in hypoxic pulmonary vasoconstriction (HPV) by acetazolamide that may have clinical utility in HAPE and possibly other pulmonary hypertensive disorders. Astonishingly, the efficacy of acetazolamide as a HPV inhibitor does not appear to be related to carbonic anhydrase inhibition, since other potent CA inhibitors have no effect on HPV either in the conscious dog or on hypoxic calcium (Ca(2+)) signalling in rat pulmonary artery smooth muscle cells, despite enzyme presence in these cells. Although we have not yet determined the mechanism of action for acetazolamide in HPV, we have ruled out actions on membrane L-type Ca(2+) channels, normoxic and hypoxic membrane potential and rho-kinase activation. Based upon these negative findings in isolated pulmonary artery smooth muscle cells and preliminary data in Ca(2+) free media we propose that acetazolamide may act at the level of Ca(2+) release from the sarcoplasmic reticulum, a process which initiates and amplifies cell membrane Ca(2+) channel opening. In further work, we have developed and will use a methylated analog of acetazolamide to yield a molecule lacking CA inhibiting activity, but which in most other respects (size, pK(a), heterocyclic ring structure, electrostatic charge distribution) is equivalent to acetazolamide. PMID- 16376159 TI - High-pressure studies of the reaction mechanism of nitric-oxide synthase. AB - Nitric-oxide synthase (NOS) generates nitric oxide from l-arginine in two reaction cycles with N(omega)-hydroxy-l-arginine as an obligate intermediate. Although much progress has been made in recent years in the elucidation of the reaction mechanism of NOS, many questions remain to be answered. The use of low temperature has been instrumental in the revelation of the mechanism of NO synthesis, particularly regarding the role of the cofactor 5,6,7,8 tetrahydrobopterin (BH4). High-pressure studies may be expected to be similarly useful, but have been very few so far. In this short review, we depict the present state of knowledge about the reaction mechanism of NO synthesis, and the role(s) BH4 plays in it. This exposition is followed by a summary of the results obtained thus far in high-pressure studies and of the conclusions that can be drawn from them. PMID- 16376161 TI - Comparison of effective dose to children and adults from dual X-ray absorptiometry examinations. AB - Dual X-ray absorptiometry (DXA) is increasingly used to measure bone density in children. If the system software does not include pediatric scan modes, then child examinations must be performed using adult scan modes that give a higher radiation dose to children than adults. This report describes a study to compare the effective dose to children and adults from DXA scans performed on the Hologic Discovery and QDR4500 models. Depth dose measurements were made using thermoluminescent dosimeters in a Rando phantom and were mapped onto the Cristy mathematical phantoms representing a 5-, 10- and 15-year-old child and an adult, and effective dose (ED) was calculated using the ICRP Publication-60 tissue weighting factors. The ED for spine (hip) examinations performed with the Express mode using the default adult scan lengths were 16.1 (9.8), 11.1 (6.7), 5.6 (3.9) and 4.4 (3.1) microSv for a 5-, 10- and 15-year-old child and adult respectively. However, if care is taken to adjust scan lengths appropriately, the child doses were reduced to 9.1 (7.4), 7.1 (5.9) and 5.0 (3.7) microSv. ED figures for the Fast and Array modes were larger by factors of 1.5 and 3 respectively. EDs for whole body scans for a 5-, 10- and 15-year-old child and adult performed on the A model (W-model) were 5.2 (10.5), 4.8 (9.6), 4.2 (8.4) and 4.2 (8.4) microSv. Using the infant whole body mode (only available on the A-model), they were 7.5 microSv for a 1-year-old and 8.9 microSv for a neonate. Although doses from child DXA examinations are low, it is still important to keep them as small as possible. DXA operators using Discovery systems can do this by using the Express scan mode, by setting appropriate values of the scan length before scan acquisition and by avoiding mistakes that lead to scans having to be unnecessarily repeated. PMID- 16376162 TI - Hydrogel-beta-TCP scaffolds and stem cells for tissue engineering bone. AB - Trabecular bone is a material of choice for reconstruction after trauma and tumor resection and for correction of congenital defects. Autologous bone grafts are available in limited shapes and sizes; significant donor site morbidity is another major disadvantage to this approach. To overcome these limitations, we used a tissue engineering approach to create bone replacements in vitro, combining bone-marrow-derived differentiated mesenchymal stem cells (MSCs) suspended in hydrogels and 3-dimensionally printed (3DP) porous scaffolds made of beta-tricalcium-phosphate (beta-TCP). The scaffolds provided support for the formation of bone tissue in collagen I, fibrin, alginate, and pluronic F127 hydrogels during culturing in oscillating and rotating dynamic conditions. Histological evaluation including toluidine blue, alkaline phosphatase, and von Kossa staining was done at 1, 2, 4, and 6 weeks. Radiographic evaluation and high resolution volumetric CT (VCT) scanning, expression of bone-specific genes and biomechanical compression testing were performed at 6 weeks. Both culture conditions resulted in similar bone tissue formation. Histologically collagen I and fibrin hydrogels specimens had superior bone tissue, although radiopacities were detected only in collagen I samples. VCT scan revealed density values in all but the Pluronic F127 samples, with Houndsfield unit values comparable to native bone in collagen I and fibrin glue samples. Expression of bone-specific genes was significantly higher in the collagen I samples. Pluronic F127 hydrogel did not support formation of bone tissue. All samples cultured in dynamic oscillating conditions had slightly higher mechanical strength than under rotating conditions. Bone tissue can be successfully formed in vitro using constructs comprised of collagen I hydrogel, MSCs, and porous beta-TCP scaffolds. PMID- 16376165 TI - Letter from the editor: the history and future of transplantation. PMID- 16376166 TI - Case of the season: intercostal hemangioma. PMID- 16376167 TI - Preoperative imaging in heart and lung transplantation in the adult. PMID- 16376168 TI - Heart transplantation imaging in the adult. PMID- 16376169 TI - Lung transplantation imaging in the adult. PMID- 16376170 TI - Liver transplantation in adults: postoperative imaging evaluation and interventional management of complications. PMID- 16376171 TI - Hepatic transplantation: pretransplant evaluation of donors and recipients. PMID- 16376172 TI - Pediatric liver transplantation. PMID- 16376173 TI - [Emergence of plasmid-mediated resistance to quinolones in Enterobacteriaceae]. PMID- 16376174 TI - Celiac disease in Tunisia: serological screening in healthy blood donors. AB - BACKGROUND: Recent epidemiological studies in Europe and in USA using antigliadin antibodies and antiendomysium antibodies for initial screening have shown that the overall prevalence of celiac disease (CD) is about 1:200 (0.5%). AIM: To screen for CD in healthy blood donors in Tunisia. PATIENTS AND METHODS: Sera from 2500 healthy blood donors (median age: 21 years, 70% men and 30% women) were screened for IgG-antigliadin antibodies and IgA-antigliadin antibodies with an enzyme-linked immunosorbent assay. All sera with positive antigliadin antibodies were tested for antiendomysium antibodies using human umbilical cord cryosections as substrate. RESULTS: Seven healthy blood donors (median age: 21 years; four men, three women) have antiendomysium antibodies. The prevalence of antiendomysium antibodies in healthy blood donors in Tunisia is 1:355 (0.28%). CONCLUSIONS: On the basis of a high specificity of the antiendomysium antibodies, it is likely that the seven blood donors identified in this study have CD. These data suggest that CD is frequent in Tunisia. PMID- 16376175 TI - High circulating levels of 7beta- and 7alpha-hydroxycholesterol and presence of apoptotic and oxidative markers in arterial lesions of normocholesterolemic atherosclerotic patients undergoing endarterectomy. AB - In previous investigations, we found that 7beta-hydroxycholesterol had potent pro apoptotic, and pro-oxidative properties. So, we asked whether the circulating level of this oxysterol was enhanced in atherosclerotic patients undergoing endarterectomy of the superficial femoral artery. To this end, 7beta hydroxycholesterol serum concentrations were determined and compared with common lipid parameters in atherosclerotic patients, and in healthy subjects. 7alpha hydroxycholesterol was simultaneously measured to evaluate the reliability of the method used for oxysterol analysis. On normal and atherosclerotic arterial fragments from patients, markers of oxidation (4-hydroxynonenal (4-HNE) adducts), and apoptosis (activated caspase-3; condensed/fragmented nuclei) were studied. Interestingly, high serum concentrations of 7beta- and 7alpha-hydroxycholesterol were found in normocholesterolemic atherosclerotic patients. However, in statin treated patients, the circulating levels of 7beta- and 7alpha-hydroxycholesterol tend towards normal values. Therefore, 7beta- as well as 7alpha hydroxycholesterol could be more appropriate markers of lipid metabolism disorders than cholesterol or LDL in normocholesterolemic patients with atherosclerosis of the lower limbs, and statins could normalize their serum concentrations. At the arterial level, apoptotic cells were mainly identified in low grade lesions and no statin effects were found on oxidation and apoptosis. PMID- 16376176 TI - [Antibiotics resistance of meticilline-resistant Staphylococcus aureus: detection of the first glycopeptides low sensibility strains in Tunisia]. AB - The adaptation of Staphylococcus aureus to the hospital environment led to the acquisition of resistance to all antibiotics available in clinical practice. The aim of this study was to investigate the methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in the F. Bourguiba hospital of Monastir (Tunisia). We determined the antibiotype of all Staphylococcus aureus strains identified. Susceptibility rates to fosfomycin, chloramphenicol, rifampicin and pristinamycin were 7%, 3%, 2% and 0%, respectively. The prevalence of MRSA was 15.5% (96 strains); their susceptible to gentamicin progressively increased. The minimum inhibitory concentration (MICs) of oxacillin, vancomycin and teicoplanin were evaluated for the 96 MRSA strains. We identified two MRSA strains (M4 and M41) showing reduced glycopeptides susceptibility. Further analysis revealed that M4 and M41 harbor the gene encoding the class S and class F proteins specific for the Panton-Valentine Leukocidin (PVL). The mecA gene was detected only in strain M41 which harbors the Staphylococcal Cassette Chromosome (SCCmec) type III. This is the first reported MRSA showing reduced susceptibility to glycopeptides in Tunisia. Regulatory surveillance of susceptibility to antibiotics is needed to reduce the morbidity and the mortality rates as well as societal costs of S. aureus infections. PMID- 16376177 TI - [Modulatory effect of antimicrobial agents or aminoacids on the oxidative burst of polymorphonuclear neutrophils triggered by formylmethionyl-leucyl phenylalanine (fMLP)]. AB - We have compared the interplay of several antimicrobial agents and aminoacids on the neutrophil respiratory burst in response to formylmethionyl-leucyl phenylalanine (fMLP), a chemoattractant. Mainly, an inhibitory effect has been observed in the penicillin family of agents and an enhancing effect in the cephalosporin family of agents. The molecules in which the sulfur numbered 1 in the 6-APA or 7-ACA nucleus was replaced by a carbon or an oxygen, had a different effect as compared with the other members of the family. The modulatory effects of ampicillin and cephalothine were not significant at a concentration lower than 10 mg/l and the effect of cephalothine looked maximum at 20-40 mg/l. If studies in cell-free systems demonstrated that the inhibitory effect of some antimicrobials could be due to a direct oxidant-scavenger activity mainly of HOCl, only hypotheses are proposed to explain the enhancing activity of the others. It could be in relation with (i) a synergistic effect upon fMLP receptor leading to an increase in H(2)O(2)/HOCl production or (ii) the generation of new oxydant products originating in cephalosporin lysis under HOCl attack, which would be able to react with luminol. The interplay of antimicrobial agents with the respiratory burst measured outside the cells probably has no therapeutic consequences because the bactericidal activity of neutrophils is achieved inside phagosomes where few agents are known to come into and where chemical conditions are different. On the opposite, in clinical use, this interplay could be interesting to study for a prevention of side effects. PMID- 16376178 TI - [Contamination of blood cultures drawn from central vein catheter and peripheral venipuncture. Prospective study of 75 pairs]. AB - OBJECTIVE: To determine the frequency of contamination, the sensitivity, specificity, and predictive values of cultures done with blood drawn through a central venous catheter or peripheral venipuncture. DESIGN: Prospective cohort study of critical ill medical surgical intensive care patients in whom samples for paired culture were drawn through a central venous catheter or peripheral venipuncture during a period of 8 months (from August 1st 2001 to Mars 31st 2002). RESULTS: During the study period, 75-paired cultures were studied. Fifteen peripheral blood cultures (20%) and 27 central blood cultures (36%) were positive and 6 peripheral blood cultures (8%) and 15 central blood cultures (20%) were contaminated (P=0.034). The organism most commonly responsible for contamination was Coagulase Negative Staphylococcus (16% of central blood cultures and 6.7% of peripheral blood cultures). The frequency of contamination was of 42.9% for blood cultures drawn through a femoral central venous catheter, of 10% for jugular, and of 19% for subclavian central venous catheter (P=0.22). For catheter draws compared with peripheral venipuncture, sensitivity was 100 and 75%, specificity was 76.2 and 90.5%, positive predictive value was 44.4 and 60% and negative predictive value was 100 and 95%. CONCLUSION: Cultures of blood drawn through a catheter are more sensible and less specific than those obtained from a peripheral venipuncture and the organism most commonly responsible for contamination of blood cultures is Coagulase Negative Staphylococcus. PMID- 16376179 TI - [DNA microarrays for gene expression profiling of breast cancer: principles and prognostic applications]. AB - Breast cancer is a major health problem in developed countries. Pathological and clinical heterogeneity, partly responsible of therapeutic failures, reflects its poorly documented complex and combinatory molecular basis. Thorough molecular typing could allow not only better tackling that diversity and improving the current prognostic classifications, but also could help in the identification of new molecular therapeutic targets. The recently developed DNA microarray technology allows the analysis of the RNA expression of several thousands of genes simultaneously in a sample. Recent studies have shown the promising prognostic impact of gene expression profiling in breast cancer by identifying new prognostic subclasses unidentifiable by conventional parameters. PMID- 16376180 TI - A fast spin echo two-point Dixon technique and its combination with sensitivity encoding for efficient T2-weighted imaging. AB - A fast spin echo two-point Dixon (fast 2PD) technique was developed for efficient T2-weighted imaging with uniform water and fat separation. The technique acquires two interleaved fast spin echo images with water and fat in-phase and 180 degrees out-of-phase, respectively, and generates automatically separate water and fat images for each slice. The image reconstruction algorithm uses an improved and robust region-growing scheme for phase correction and achieves consistency in water and fat identification between different slices by exploiting the intrinsic correlation between the complex images from two neighboring slices. To further lower the acquisition time to that of a regular fast spin echo acquisition with a single signal average, we combined the fast 2PD technique with sensitivity encoding (SENSE). Phantom experiments show that the fast 2PD and SENSE are complementary in scan efficiency and signal-to-noise ratio (SNR). In vivo data from scanning of clinical patients demonstrate that T2-weighted imaging with uniform and consistent fat separation, including breath-hold abdominal examinations, can be readily performed with the fast 2PD technique or its combination with SENSE. PMID- 16376181 TI - Implementation of a rapid inversion-prepared dual-contrast gradient echo sequence for quantitative dynamic contrast-enhanced magnetic resonance imaging of the human prostate. AB - The first step in quantitative pharmacokinetic modeling is to determine the arterial input function (AIF) by deriving the contrast medium (CM) concentration from an appropriate imaging sequence by monitoring changes in either the amplitude or the phase signal of an accommodative artery. The bolus passage is best detected on T2- or T2*-weighted images, while extravasation is best assessed on T1-weighted images. Here, an imaging sequence is used that employs a parallel acquisition technique for the interleaved acquisition of an inversion-prepared T1 weighted image and a T1/T2*-mixed-weighted image for determination of the AIF. The sequence was applied in six patients with prostate cancer. A method is presented for quantifying the AIF derived from the signal intensity-time courses of both the T1/T2*-mixed-weighted and the T1-weighted image. Furthermore, in some patients the signal intensity-time course of the T1-weighted image exhibits flow induced signal modulations. To reduce the effect of this flow-related signal enhancement the corresponding phase information was used. The sequence presented here has the potential to improve the quantification of the AIF at all time points and pharmacokinetic modeling of the CM dynamics of the prostate. PMID- 16376182 TI - Assessment of pulmonary air trapping and obstruction in expiration: an experimental MRI study. AB - PURPOSE: The aim of this experimental study was to evaluate the potential of a simple expiration technique by means of magnetic resonance imaging (MRI) in an animal model to detect pulmonary air-trapping areas after artificial bronchial obstruction. MATERIAL AND METHODS: Sixteen pigs were evaluated by means of a modified T1-weighted FLASH with fat saturation in respiratory arrest (TR=4.6 ms, TE=1.8 ms, alpha=10 degrees, S.D.=3-5 mm). A measurement of the signal intensity (SI) in the peripheral lung tissue was made in both inspiration and expiration before and after inhalation of 2 ml of 0.5% acetylcholine to simulate a bronchial obstruction. A final measurement of the lung SI was also made after bronchospasmolytic induction through salbutamol (beta2-mimetic bronchodilator). RESULTS: In expiration, a mean SI increase in peripheral lung tissue of about 183% was seen in comparison to inspiration (mean SI increase of 11-32). After inhalation of 0.5% acetylcholine, the expirational signal increase in peripheral lung tissue was only 114% of the original SI. The expirational signal homogeneity decreased after inhalation of acetylcholine. After inhalation of salbutamol, the lung tissue signal elevation in expiration was 193%. CONCLUSION: We interpret the low expiratory signal elevation after acetylcholine inhalation as a result of an air-trapped bronchial constriction in certain areas. The simple expiratory technique in an animal model showed that it is suitable to demonstrate obstructive air trapping using MRI. PMID- 16376183 TI - Spinal effects of acupuncture stimulation assessed by proton density-weighted functional magnetic resonance imaging at 0.2 T. AB - Signal changes can be detected by proton density-weighted functional imaging in both the brain and the spinal cord. These are attributed to changes in extravascular water proton (signal enhancement by extravascular protons) density during neuronal activation. In this study, we used this technique to detect correlations between acupoint stimulation and neural activity in the spinal cord. Stimulation of acupoints associated with treatment of sensorimotor deficits (LI4 and LI11) was performed on 11 volunteers. During stimulation, 8 of the 11 subjects had consistent functional activations in C6/C7. A bilateral activation pattern was common. Our findings show that acupoint stimulation modulates activity in the spinal cord. PMID- 16376184 TI - Relaxivities of human liver and spleen ferritin. AB - Ferritin, the iron-storing protein of mammals, is known to darken T2-weighted magnetic resonance images. This darkening can be used to noninvasively measure an organ's iron content. Significant discrepancies exist between T2 data obtained with ferritin-containing tissues and with aqueous solutions of horse spleen ferritin (HSF). The NMR properties of stable human ferritin have never been studied in aqueous solutions. Relaxometry results on human liver and spleen ferritin are reported here, showing that the relaxation induced in aqueous solutions by human ferritins is comparable to that induced by HSF. As a consequence, the differences between ferritin-containing human tissues and ferritin solutions cannot be attributed to different NMR properties of human and horse ferritins, but probably to a clustering of the protein in vivo. PMID- 16376185 TI - Potential of magnetic resonance spectroscopy to detect metastasis in axillary lymph nodes in breast cancer. AB - Focused pathological evaluation of axillary lymph nodes in breast cancer is gaining importance. Nuclear magnetic resonance (NMR) spectroscopy that assesses the whole of the specimen has the potential in evaluating micrometastases. The biochemical changes associated with breast cancer metastases in axillary nodes by in vitro NMR and its use in the detection of axillary metastases in a clinical setting in comparison with conventional histopathology is presented in this study. Eighty-eight lymph nodes obtained from 30 patients with breast cancer were investigated. Histopathology revealed metastases in 20 nodes from 11 patients, while in vitro NMR spectroscopy revealed metastases in 22 nodes. Out of these 22 nodes, 16 were the same, which showed metastases on histopathology, while 6 nodes have shown metastases only on in vitro magnetic resonance spectroscopy (MRS). These 6 nodes with suspicion of metastases on MRS were subjected to reevaluation with serial sectioning and immunohistochemistry, but no additional metastases were revealed. Forty metabolites could be identified from the MR spectrum of lymph nodes. The levels of the glycerophosphocholine-phosphocholine (GPC-PC), choline, lactate, alanine and uridine diphosphoglucose were elevated significantly in nodes with metastases. In addition, the intensity ratio of GPC PC/threonine (Thr) was higher in nodes with metastases, and using this as marker, MRS detected the axillary metastases with a sensitivity, specificity and accuracy of 80%, 91% and 88%, respectively. Neoadjuvant chemotherapy (NACT) lowered the concentrations of GPC-PC and GPC-PC/Thr ratio. The accuracy of MRS in detecting metastases was 75% in patients who received NACT (n=9) as compared to 96% in those who did not (n=21). Our results demonstrate the potential of in vitro MRS in characterizing the metabolite profile of the axillary nodes with breast cancer metastases. It detected axillary metastases with reasonable accuracy and can be complementary to histopathological evaluation and immunohistochemistry. PMID- 16376186 TI - Trabecular bone volume fraction measurements of a large number of subjects using a compact MRI. AB - Trabecular bone volume fraction (TBVF) and speed of sound (SOS) were measured for the right calcanei of 416 female volunteers. The TBVF was measured with a compact MRI developed in our laboratory. The SOS was measured with a commercial quantitative ultrasound system. It was observed that the correlation coefficient between TBVF and SOS and that between TBVF and age varied depending on the location of region of interest (ROI) in the calcaneus. As a result, an optimum circular ROI with a diameter of 20 mm was determined so that the correlation coefficients were maximized. In the optimum ROI, transverse relaxation time (T2) of the bone marrow protons of the calcaneus was found to be concentrated in a narrow range over the subjects. This result suggested that a 50% scan time reduction in the TBVF measurements could be made by skipping the T2 correction procedure. PMID- 16376187 TI - Ferrite-enhanced MRI monitoring in hyperthermia. AB - In an MRI hyperthermia hybrid system, T1 changes are investigated for monitoring thermal therapy at 0.2 T. The water bolus, which is needed for power transmission and cooling of the skin, limits MR image quality by signal compression and artifacts. Superparamagnetic ferrofluid in different concentration was investigated with MR relaxometry and MRI methods. We found that using ferrofluid in a low concentration of 70-90 ppm magnetite the water signal can be suppressed without susceptibility artifacts. With our method of signal suppression, a significant improvement of spatial and temporal resolution is possible. The ferrofluid is stable and allows RF heating at 100 MHz. This method of signal extinction may also be useful for other experimental setups where suppression of water is necessary. PMID- 16376188 TI - Septal repair implants: evaluation of magnetic resonance imaging safety at 3 T. AB - Specialized implants are used for transcatheter closure of septal defects, including atrial and ventricular septal defects, and patent foramen ovale. These metallic devices may pose a risk to patients undergoing magnetic resonance imaging (MRI) procedures especially if performed at 3 T. Therefore, this investigation evaluated MRI safety at 3 T for septal repair implants (CardioSEAL Septal Repair Implant and STARFlex Septal Repair Implant, NMT Medical, Boston, MA, USA) by characterizing magnetic field interactions, heating and artifacts. These implants exhibited minor magnetic field interactions; heating was not excessive (+0.5 degrees C); and artifacts will only create a problem if the area of interest is in the same area as or near these devices. Thus, the findings indicated that it would be safe for a patient with these implants to undergo MRI at 3 T or lower. Importantly, because of the minor magnetic field interactions, MRI may be performed immediately after implantation. PMID- 16376189 TI - Experimental development of a petal resonator surface coil. AB - A surface coil for MRI was designed and built based on the principles of the petal resonator proposed by Mansfield [J Phys D Appl Phys 21 (1988) 1643]. This resonator coil design was named the petal resonator surface (PERES) coil and is composed of an eight-petal coil array and a central circular coil. A minimum separation of three times the petal coil radius is necessary to significantly decrease the mutual inductance. An analytical function for the PERES Signal-to noise ratio (SNR) is obtained based on the quasistatic method. Theoretical plots of SNR enhancement yielded 26% and 35% more SNR over the circular coil and phased array coils. Imaging experiments were first performed using a spectroscopy phantom on a 1.5-T commercial imager. Subsequently, brain images of healthy volunteers were obtained. Clinical MR imager compatibility allows this resonator coil to be used with conventional pulse sequences and imaging protocols. This coil design offers a new alternative to existing surface coils because it significantly increases the SNR. PMID- 16376190 TI - Factors associated with abdominal compartment syndrome complicating endovascular repair of ruptured abdominal aortic aneurysms. AB - BACKGROUND: Endovascular treatment of ruptured abdominal aortic aneurysms (r AAAs) has the potential to offer improved outcomes. As our experience with endovascular repair of r-AAA evolved, we recognized that the development of abdominal compartment syndrome (ACS) led to an increase in morbidity and mortality. We therefore reviewed our experience to identify risk factors associated with the development of ACS. METHODS: From January 2002 to December 2004, 30 patients underwent emergent endovascular repair of r-AAA by using commercially available stent grafts. All patients who developed ACS underwent emergent laparotomy. Physiological and clinical parameters were analyzed between patients with and without ACS after endovascular r-AAA repair. RESULTS: Over the past 3 years, 30 patients underwent endovascular r-AAA repair, and 6 (20%) patients developed ACS. Patients with ACS had a higher incidence of the need for aortic occlusion balloon (67% vs 12%; P = .01), a markedly longer activated partial thromboplastin time (128 +/- 84 seconds vs 49 +/- 31 seconds; P = .01), a greater need for blood transfusion (8 +/- 2.5 units vs 1.8 +/- 1.7 units; P = .08), and a higher incidence of conversion to aortouni-iliac devices because of ongoing hemodynamic instability and an inability to expeditiously cannulate the contralateral gate (67% vs 8%) when compared with patients without ACS. The mortality was significantly higher in the patients with ACS (67%; 4 of 6) compared with patients without ACS (13%; 3 of 24; P = .01). CONCLUSIONS: ACS is a potential complication of endovascular repair of r-AAA and negatively affects survival. Factors associated with the development of ACS include (1) use of an aortic occlusion balloon, (2) coagulopathy, (3) massive transfusion requirements, and (4) conversion of bifurcated stent grafts into aortouni-iliac devices. We recommend that, after endovascular repair of r-AAA, these patients undergo vigilant monitoring for the development of ACS. PMID- 16376191 TI - Midterm clinical success and behavior of the aneurysm sac after endovascular AAA repair with the Excluder graft. AB - OBJECTIVE: Recent studies have reported different sac behavior after endovascular repair of abdominal aortic aneurysms, depending on the endografts. This study was designed to evaluate mid-term outcome and sac behavior after treatment with the Gore Excluder stent-graft. METHODS: Between June 1999 and January 2005, 109 selected patients with suitable anatomy were treated electively for abdominal aortic aneurysm with the Excluder stent graft. Data were prospectively collected in a computerised database and included demographics, details of the aortoiliac anatomy, procedural and clinical success, and postoperative complications. Postoperative sac size and the presence of endoleaks were assessed with computed tomography scans obtained at 1, 6, and 12 months, and yearly thereafter. All diameter measures in patients followed for > 1 year (84.4%) were analyzed. RESULTS: Assisted primary technical success was achieved in 108 cases (99.1%). No type I endoleaks and 12 (11.1%) type II endoleaks were recorded < or = 1 month from the procedure. Mean follow-up was 29.6 +/- 16.1 months. We recorded one new onset type IA endoleak, complete resolution of five type II endoleaks, and eight new-onset type II endoleaks. The overall prevalence of type II endoleaks was 14%. Shrinkage at 1, 2, 3, and 4 years was observed in 20.7%, 30.5%, 38.9%, and 36.8% of cases. The presence of type II endoleak influenced the trend of aneurysm size throughout the 4 years. Aneurysms without endoleak shrank more than aneurysms with type II endoleak (P < .0001). We observed two cases of sac enlargement due to the presence of endoleaks. No cases of endotension with sac enlargement, late open conversion, or aneurysm-related deaths were observed. Unchanged aneurysmal sacs remained stable during follow-up, with no adverse events. CONCLUSIONS: Endovascular treatment with the Excluder device in selected patients produces low rates of shrinkage, but this peculiar sac behavior does not affect mid-term clinical success. PMID- 16376192 TI - Amplatzer Vascular Plug to occlude the internal iliac arteries in patients undergoing aortoiliac aneurysm repair. AB - PURPOSE: The purpose of this report is to evaluate the use of conventional coils and the Amplatzer Vascular Plug, a type of nitinol-based self-expanding device, to occlude the internal iliac artery in patients undergoing aortoiliac or common iliac aneurysm endograft repair, or both. METHODS: Between August and December 2004, in preparation for endograft repairs of aortoiliac or common iliac artery aneurysms, or both, at a community hospital system, five patients underwent the occlusion of the internal iliac artery with an Amplatzer Vascular Plug to prevent endoleak. During the preceding 12 months, the conventional coil embolization of the internal iliac artery was used for the same purpose in 10 patients. RESULTS: In five patients undergoing the Amplatzer Vascular Plug occlusion of the internal iliac artery, precise deployment at the origin of the artery was achieved. Complete and precise occlusion was confirmed angiographically, and only one device was used for each internal iliac artery. Two patients reported mild buttock claudication 2 weeks after occlusion, which resolved completely by 6 and 8 weeks, respectively. A type II endoleak from the inferior mesenteric artery developed in one patient. In the previous 10 patients, 11 internal iliac arteries were treated with conventional coils. Subsequent repeat coil embolization was required for three patients. The procedural complications in this second group included one case of coil embolization into the superficial femoral artery and one into the common iliac artery; both errant coils were retrieved successfully by endovascular techniques. An average of 7 +/- 3.4 (mode of 5) coils were used for each internal iliac artery. Three cases of buttock claudication occurred after the unilateral internal iliac artery occlusion in this group and did not resolve. No evidence of ischemic bowel, buttock necrosis, or sexual dysfunction was observed in either group. The estimated average cost to occlude one internal iliac artery was 375 dollars for Amplatzer Vascular Plugs and 3,500 dollars for conventional coils. CONCLUSIONS: The Amplatzer Vascular Plug allows for a cost effective method to occlude the internal iliac artery in patients undergoing endograft repairs of aortoiliac aneurysms. The use of a single device with a precise placement at the origin of the artery minimizes cost and avoids ischemic complications. PMID- 16376193 TI - Endovascular repair of lesions involving the descending thoracic aorta. AB - BACKGROUND: Vascular lesions involving the thoracic aorta are often life threatening conditions that carry significant morbidity and mortality with traditional open surgical repair. Preliminary results suggest that endovascular therapy is an effective and possibly advantageous treatment for diseases of the descending thoracic aorta. METHODS: Between October 2000 and May 2004, 50 consecutive patients underwent endovascular stent-grafting of lesions involving the descending thoracic aorta. Attempted stent-graft deployment was performed electively in 39 patients and emergently in 11. The pathology of electively treated aortic lesions included degenerative/atherosclerotic aneurysms (n = 24), pseudoaneurysms (n = 11), aortic dissections (n = 2), and penetrating ulcers (n = 2). Emergently treated aortic lesions were for acute rupture due to infectious (mycotic) aneurysms (n = 4), atherosclerotic/degenerative aneurysms (n = 3), acute type B dissections (n = 2), and acute transections (n = 2). Devices used include Talent (n = 45), AneuRx aortic cuffs (n = 2), custom-fabricated Gianturco Dacron grafts (n = 2), and a modified Cook-Zenith abdominal aortic graft (n = 1). Follow-up was performed at 1-month, 6-months, 1-year, and annually thereafter. RESULTS: Primary technical success, defined as successful deployment and exclusion of the lesion without evidence of type I or type III endoleak, was achieved in 48 (96%) of 50 patients. In one patient, the procedure was terminated due to inability to access the iliac vessels. In another patient, a type III endoleak was observed at the completion of the primary procedure that required deployment of an additional stent-graft component 2 months later. Of the 49 patients who received endografts, seven underwent secondary procedures to correct endoleaks, with five of these seven requiring the deployment of additional endovascular stent-graft components. Major complications included four in hospital deaths, with three of these occurring in patients treated emergently. Additionally, respiratory failure (n = 6), multisystem organ failure (n = 2), cerebrovascular accident (n = 2), retroperitoneal hematoma (n = 2), acute renal insufficiency (n = 1), and pulmonary embolus (n = 1) were also observed. The overall endoleak rate was 20%, with five primary (< or = 30 days) and five secondary (> 30 days) endoleaks observed. Five of the endoleaks were treated with the deployment of one or more additional endovascular stent-graft components. Two of the endoleaks were treated with endovascular balloon remolding. Mean follow-up was 271 days. There were no aneurysm ruptures or aneurysm-related deaths. CONCLUSIONS: Endovascular treatment of vascular lesions involving the descending thoracic aorta can be safely performed with low morbidity in high-risk patients. Endovascular repair may become an attractive alternative for the treatment of a wide range of pathology along this vascular territory. PMID- 16376194 TI - Carotid atherosclerotic plaques in patients with transient ischemic attacks and stroke have unstable characteristics compared with plaques in asymptomatic and amaurosis fugax patients. AB - INTRODUCTION: Atherosclerotic carotid artery disease is responsible for a variety of clinical presentations, ranging from asymptomatic to cerebral ischemic events. Considering the upcoming use of noninvasive imaging modalities, plaque characteristics could serve as a marker in the selection of patients eligible for carotid endarterectomy (CEA). This would be more likely if characteristics corresponded with clinical manifestations and were predictive of future events. In this study, we hypothesized that plaque characteristics correlate with the clinical presentation of carotid artery disease. METHODS: We included 404 patients undergoing a carotid endarterectomy (CEA). Ipsilateral clinical symptoms and duplex measurements were recorded. Patients could be asymptomatic (23.5%) or symptomatic with stroke (26.5%), transient ischemic attack (TIA) (36.1%), or amaurosis fugax (AFX) (13.9%). Plaques were stained and semi-quantitatively analyzed for the presence of macrophages, smooth muscle cells, collagen, calcifications, and thrombus. Plaques were categorized in three phenotypes by their overall presentation and the amount of fat. In addition, plaque matrix metalloproteinase (MMP) activity and cytokines expressions were measured. RESULTS: Fibrous, fibro-atheromatous, and atheromatous plaques were observed in 30.2%, 35.6%, and 34.2%, respectively. Atheromatous plaques were more prevalent in patients with stroke and TIA compared with asymptomatic patients or patients with AFX (P = .001). Collagen staining was less evident in patients with TIA and stroke compared with asymptomatic patients or patients with AFX (P < .001). Plaques of patients with TIA and stroke showed significantly higher activity levels of MMP-8 and MMP-9 and higher levels of interleukin-8 compared with asymptomatic and AFX patients. CONCLUSION: Plaque phenotype of patients with TIA is comparable to that of patients with stroke; whereas, the plaque phenotype of patients with AFX resembles the plaque phenotype of asymptomatic patients. Follow up studies should be encouraged to determine whether plaque characteristics visualized by imaging techniques might help to identify patients most likely to benefit from CEA. PMID- 16376195 TI - Closure of the arteriotomy after carotid endarterectomy: patch type is related to intraoperative microemboli and restenosis rate. AB - OBJECTIVE: Patch closure after carotid endarterectomy (CEA) improves clinical outcome compared with primary closure. Whether there are differences in outcome between various patch materials is still not clear. The objective of this retrospective study was to investigate whether a relationship exists between the patch type and the number of microemboli as registered during CEA by transcranial Doppler imaging, the clinical outcome (transient ischemic attack and cerebrovascular accident), and the occurrence of restenosis. METHODS: We included 319 patients who underwent CEA. Intraoperative microembolus registration was performed in 205 procedures. Microembolization was recorded during four different periods: dissection, shunting, clamp release, and wound closure. The decision to perform primary closure or to use a patch for the closure of the arteriotomy was made by the surgeon, and Dacron patches were used when venous material was insufficient. Cerebral events were recorded within the first month after CEA, and duplex scanning was performed at 3 months (n = 319) and 1 year (n = 166) after CEA. A diameter reduction of more than 70% was defined as restenosis. RESULTS: Primary, venous, and Dacron patch closures were performed in 83 (26.0%), 171 (53.6%), and 65 (20.4%) patients, respectively. Primary closure was significantly related to sex (Dacron patch, 35 men and 30 women; venous patch, 108 men and 63 women; primary closure, 72 men and 11 women; P < .001). The occurrence of microemboli during wound closure was also related to sex (women, 2.5 +/- 0.6; men, 1.0 +/- 0.2; P = .01). Additionally, during clamp release, Dacron patches were associated with significantly more microemboli than venous patches (11.1 +/- 3.4 vs 4.0 +/- 0.9; P < .01), and this difference was also noted during wound closure (3.1 +/- 0.9 vs 1.4 +/- 0.4; P < .05). Transient ischemic attacks and minor strokes after CEA occurred in 5 (2.4%) of 205 and 6 (2.9%) of 205 procedures, respectively, and the degree of microembolization during dissection was related to adverse cerebral events (P = .003). In contrast, the type of closure was not related to immediate clinical adverse events. However, primary closure and Dacron patches were associated with an increase in the restenosis rate compared with venous patches: after 400 days, the restenosis rate for Primary closure was 11%, Dacron patch 16%, and venous patch 7% (P = .05; Kaplan Meier estimates). CONCLUSIONS: Microemboli are more prevalent during clamp releases and wound closure when Dacron patches are used. Additionally, the observed differences in embolization noted by patch type were mainly evident in women. However, the use of Dacron patches was not related to immediate ischemic cerebral events but was associated with a higher restenosis rate compared with venous patch closure. This suggests that venous patch closure may be preferred for CEA. PMID- 16376196 TI - Mini skin incision for carotid endarterectomy (CEA): a new and safe alternative to the standard approach. AB - PURPOSE: Patients requiring surgery are naturally attracted to shorter incisions because they tend to cause less pain and are esthetically more appeasing. To substantially shorten the length of standard skin incisions (4 to 7 inches) for carotid endarterectomy (CEA), we used preoperative duplex scanning to outline the carotid bifurcation as well as to determine the extent of disease in both the internal and common carotid arteries. METHODS: During the last 21 months, 265 consecutive primary CEAs were performed in 253 patients (mean age 72 +/- 10 years) at a single institution. Of these, 142 were men (56%). Hypertension, coronary artery disease, diabetes mellitus, smoking, and chronic renal failure were present in 81%, 44%, 43%, 28%, and 19% of the patients, respectively. Neurologically asymptomatic patients accounted for 71% of the cases. All patients received general anesthesia. Duplex-assisted skin markings of the diseased carotid artery were performed after proper patient positioning on the operating table. Synthetic patches were routinely used, and intraluminal shunts were deemed necessary by low stump pressures in 64 cases (24%). Completion duplex scanning was performed in all cases. RESULTS: The length of the longitudinal skin incision varied from 0.8 to 3.5 inches (average 1.4 +/- 0.5 inches). It was < or = 1 inch in 56 cases (21%), 1.1 to 1.5 inches in 110 (42%), 1.6 to 2 inches in 85 (32%), and 2.1 to 3.5 inches in the remaining 14 cases (5%). Intraluminal shunts were required in 9 (16%), 18 (16%), 29 (34%), and 8 (57%) of the cases, respectively. Incisions were longer in cases requiring an indwelling shunt (1.6 +/- 0.6 inches vs 1.4 +/- 0.4 inches) (P < .0001). The average patch length was 1.3 +/- 0.3 inches (range, 0.7 to 2.6 inches). The skin incision averaged 1.54 +/- 0.45 inches for the first 133 cases and 1.35 +/- 0.45 inches for the remaining 132 cases (P < .0001). Technical defects occurred in 10 cases (3.8%). The overall incidence of ipsilateral stroke and death was 1.9% and 0%. There were no technical defects or strokes in patients with the shortest incisions (< or = 1 inch). Overall, there were three transitory peripheral nerve injuries (1.1%). A comparative analysis with 265 consecutive CEAs performed immediately before this series without duplex-assisted skin markings revealed no significant differences in age (71 +/- 11 years), incidence of neurologically symptomatic patients (26%), sex (60% men), shunt use (24%), and major technical defects (3%). Also, postoperative transitory peripheral nerve injury (0.8%), stroke (0%), and death (0%) were not significantly different from the duplex-assisted group. It is of interest to note that none of the former cases was performed with a skin incision < or = 2 inches. CONCLUSION: Most CEAs (95%) can safely be performed with < or = 2-inch skin incisions. Pre-CEA duplex-assisted skin marking is a novel approach that confirms the side of the operation, localizes the disease, and minimizes the magnitude of the operation via shorter, more esthetically pleasing incisions. PMID- 16376197 TI - Assessment of short-term outcomes for protected carotid angioplasty with stents using recent evidence. AB - BACKGROUND: Carotid artery stenosis is an important risk factor and etiology of stroke. Carotid endarterectomy (CEA) is the gold standard for the treatment of carotid artery stenosis; however, there are potential benefits to adopting the use of carotid artery stenting (CAS) with protection devices. A number of large protected CAS (PCAS) trials are underway, but final results are still several years away. In the interim, numerous PCAS studies have recently been published, and the aim of this study was to combine the published results and examine the outcomes and safety of PCAS. METHODS: Electronic, manual, and bibliographic searches of PubMed and PreMedline were conducted. Proportion differences were calculated for the periprocedural (30-day) outcomes of any stroke and any stroke or death. RESULTS: More than 400 articles were identified. Only 26 studies met the inclusion criteria, resulting in the inclusion of 2,992 patients treated with PCAS. Within this patient group, the pooled perioperative PCAS rate of any type of stroke was 2.4% +/- 0.3% (95% confidence interval [CI]). The 30-day minor stroke rate was 1.1% +/- 0.2% (95% CI), and the 30-day major stroke rate was 0.6% +/- 0.2% (95% CI). The 30-day mortality rate was 0.9% +/- 0.4% (95% CI). CONCLUSION: This study demonstrates relatively low rates of reported perioperative adverse events in PCAS. Selective use of PCAS to treat carotid artery stenosis in those at highest risk for surgical complications is appropriate until the randomized trials of CEA vs PCAS provide concurrent short- and long-term outcome data. PMID- 16376198 TI - Incidence and risk factors for medical complications after carotid artery stenting. AB - OBJECTIVE: Carotid angioplasty and stenting (CAS) is being evaluated as an alternative to carotid endarterectomy for the treatment of carotid artery stenosis; however, to date little is known about the incidence of medical complications after CAS. The goal of this study was to determine the frequency of, and to identify potential clinical risk factors for, the development of medical complications after CAS. METHODS: Medical complications that occurred < or = 30 days after CAS in 327 consecutive patients (241 men, 86 women; mean age, 69 +/- 9 years; range, 45 to 90 years) treated for symptomatic (n = 182, 56%) or asymptomatic (n = 145, 44%) carotid artery stenosis were recorded. The effect of clinical characteristics on the subsequent development of medical complications was analyzed by logistic regression. RESULTS: Fifty-one patients (15%) had 62 medical complications: 3 (0.9%) myocardial infarctions, 3 (0.9%) cardiac arrhythmias, 4 (1.2%) episodes of angina pectoris, 3 (0.9%) episodes of symptomatic hypertension, 16 (4.9%) episodes of symptomatic hypotension, 10 (3.1%) chest infections, 9 (2.7%) had periods of confusion, 5 (1.5%) had urinary retention, and 9 (2.7%) urinary tract infections. One chest infection was fatal and 16 complications prolonged the intensive care unit monitoring period > 24 hours. Advanced age (odds ratio [OR], 1.1; 95% confidence interval [CI], 1.05 to 1.14) and a symptomatic carotid stenosis (OR, 2.1; 95% CI, 1.07 to 4.1) independently predicted the occurrence of medical complications. CONCLUSION: Although life-threatening or fatal non-neurologic events were uncommon in this series, the overall incidence of medical complications after CAS might be higher than currently anticipated. Older and symptomatic patients are at the highest risk, and these subgroups should be monitored closely. PMID- 16376200 TI - Duplex-guided balloon angioplasty and stenting for femoropopliteal arterial occlusive disease: an alternative in patients with renal insufficiency. AB - BACKGROUND: The technique of balloon angioplasty of infrainguinal arteries requires standard arteriography and fluoroscopic guidance. We attempted to perform this procedure under duplex guidance to avoid the use of nephrotoxic contrast material and radiation exposure in patients with renal insufficiency. METHODS: Over 16 months, 28 patients (17 men) with serum creatinine levels of 1.5 mg/dL or more underwent 37 lower extremity duplex-guided balloon angioplasties at our institution. Ages ranged from 58 to 92 years (mean +/- SD, 74 +/- 9 years). Disabling claudication was the indication in 24 cases (65%), and critical ischemia, in 13 cases. Preoperative duplex arterial mapping showed severe superficial femoral artery and/or popliteal artery stenoses in all cases. No arterial occlusions were treated in this series. Seven procedures (19%) were performed for restenosis. The ipsilateral common femoral artery was cannulated in 32 cases (86%), and the contralateral common femoral artery, in 5 cases (14%), under direct duplex visualization. Contralateral common iliac artery cannulations were performed with the help of fluoroscopy. By using sonographic visualization, a guidewire was directed into the origin of the superficial femoral artery, across the diseased segment, and into the popliteal artery. The diseased segment was then balloon-dilated. Balloon diameter and length were chosen according to arterial measurements obtained by duplex scan. Plaque dissections and recoils causing stenosis of 30% or more, a peak systolic velocity ratio of 2 or more, or both were stented under duplex guidance. Arterial duplex examinations and ankle/brachial indexes were obtained before hospital discharge, within 1 month after the procedure, and every 3 months thereafter. RESULTS: Thirty-day survival was 100%. Local complications included one open exploration for expanding hematoma. Technical success was achieved in all cases. Placement of intraluminal stents was deemed appropriate in 23 (62%) of 37 cases. The 1-month patency and limb salvage rates were 100%. Preprocedure and postprocedure ankle/brachial indexes ranged from 0.3 to 0.9 (mean +/- SD, 0.64 +/- 0.16) and 0.64 to 1.2 (mean +/- SD, 0.92 +/- 0.15), respectively (P < .0001). CONCLUSIONS: Duplex-guided balloon angioplasty seems to be a safe and effective technique that allows renal patients to experience continued limb salvage and relief from claudication without the risk of developing dye-induced acute renal failure. Other advantages include direct visualization of the puncture site, accurate selection of the proper size of balloon and stent, confirmation of the adequacy of the technique by hemodynamic and imaging parameters, and avoidance of radiation. Although this technique holds considerable potential, longer follow-up will help to fully evaluate its broader applicability. PMID- 16376201 TI - Duplex-guided balloon angioplasty and subintimal dissection of infrapopliteal arteries: early results with a new approach to avoid radiation exposure and contrast material. AB - OBJECTIVE: Balloon angioplasties of stenotic or occluded infrapopliteal arteries may be helpful in selected high-risk patients threatened with limb loss. Thus far, these procedures have demanded fluoroscopy and the injection of potentially nephrotoxic contrast material. Herein, we proposed a new alternative to avoid the harmful effects of radiation exposure and the risk of acute renal failure. METHODS: Over the last 16 months, 30 patients (57% male) aged 74 +/- 9 years (mean +/- SD) had a total of 52 attempted balloon angioplasties of the infrapopliteal arteries in 32 limbs under duplex guidance. Indications for the procedure were critical ischemia in 20 limbs (63%), including rest pain, ischemic ulcers, and gangrene in 4 (13%), 10 (31%), and 6 (19%) limbs, respectively. Severe disabling claudication was an indication in the remaining 12 limbs (37%). All patients had concomitantly performed balloon angioplasties of the superficial femoral and popliteal arteries (28 cases) or the popliteal artery alone (4 cases). Balloon angioplasty of the infrapopliteal arteries was performed as an adjunct to improve runoff. Hypertension, diabetes, renal insufficiency, smoking, and coronary artery disease were present in 77%, 73%, 50%, 47%, and 37% of cases, respectively. There were 42 cases (81%) with infrapopliteal arterial stenoses (25 tibioperoneal trunks, 9 peroneal arteries, 4 anterior tibial arteries, and 4 posterior tibial arteries) in 26 limbs. The remaining 10 cases (19%) had infrapopliteal arterial occlusions (4 tibioperoneal trunks, 5 peroneal arteries, and 1 anterior tibial artery) in 6 limbs. All these cases were combined with more proximal endovascular procedures (21 femoropopliteal stenoses and 11 femoropopliteal occlusions). All patients had preprocedure duplex arterial mapping and ankle/brachial index (ABI) measurement. Local anesthesia with light sedation was used in all cases. The common femoral artery was cannulated under direct duplex visualization. Still under duplex guidance, a guidewire was directed into the proximal superficial femoral artery and distally, beyond the infrapopliteal diseased segment. The diseased segment was then balloon-dilated. Balloon diameter and length were chosen according to the arterial measurements obtained by duplex guidance. Completion duplex examinations were performed and postprocedure ABIs were obtained in all cases. RESULTS: Although the overall technical success was 94% (49/52 cases), it was 95% for those with stenoses (40/42 cases) and 90% for those with occlusions (9/10 cases; P < .5). Intraoperative thrombosis occurred in three infrapopliteal cases (two tibioperoneal trunks and one peroneal artery) and in one popliteal artery. All four cases were successfully managed with intra-arterial infusion of thrombolytic agents under duplex guidance. Overall, the preprocedure and postprocedure ABIs ranged from 0.4 to 0.8 (mean +/- SD, 0.58 +/- 0.15) and 0.7 to 1.1 (mean +/- SD, 0.9 +/- 0.16), respectively (P < .0001). Twenty-two (88%) of 25 patients experienced a significant (> 0.15) postoperative ABI increase. Overall 30-day survival and limb salvage rates were 100%. CONCLUSIONS: The proposed technique eliminates the need for radiation exposure and the use of contrast material, and it seems to be an effective alternative approach for the treatment of infrapopliteal occlusive disease. Additional advantages include accurate selection of the proper size of balloon and confirmation of the adequacy of the technique by hemodynamic and imaging parameters. PMID- 16376202 TI - Upper- vs lower-limb aerobic exercise rehabilitation in patients with symptomatic peripheral arterial disease: a randomized controlled trial. AB - OBJECTIVES: To investigate the effects of a 24-week program of upper- and lower limb aerobic exercise training on walking performance in patients with symptomatic peripheral arterial disease (PAD) and to study the mechanisms that could influence symptomatic improvement. METHODS: After approval from the North Sheffield Local Research Ethics Committee, 104 patients (median age, 69 years; range, 50 to 85 years) with stable PAD were randomized into an upper- or lower limb aerobic exercise training group (UL-Ex or LL-Ex), or to a nonexercise training control group. Training was performed twice weekly for 24 weeks at equivalent relative exercise intensities. An incremental arm- and leg-crank test (ACT and LCT) to maximum exercise tolerance was performed before and at 6, 12, 18, and 24 weeks of the intervention to determine peak oxygen consumption (VO(2)). Walking performance, defined as the claudicating distance (CD) and maximum walking distance (MWD) achieved before intolerable claudication pain, was assessed at the same time points by using a shuttle-walk protocol. Peak blood lactate concentration, Borg ratings of perceived exertion (RPE) and pain category ratio (CR-10) were recorded during all assessments. RESULTS: CD and MWD increased over time (P < .001) in both training groups. At 24 weeks, CD had improved by 51% and 57%, and MWD had improved by 29% and 31% (all P < .001) in the UL-Ex and LL Ex groups, respectively. An increase in peak heart rate at MWD in the UL-Ex group (109 +/- 4 vs 115 +/- 4 beats/min; P < .01) and LL-Ex group (107 +/- 3 vs 118 +/- 3 beats/min; P = .01) was accompanied by an increase in the amount of pain experienced in both groups (P < .05), suggesting that exercising patients could tolerate a higher level of cardiovascular stress and an increased intensity of claudication pain before test termination after training. Patients assigned to exercise training also showed an increase in LCT peak VO2 at the 24-week time point in relation to baseline (P < .01) and control patients (P < .01), whereas ACT peak VO2 was only improved in the UL-Ex group (P < .05). CONCLUSIONS: Our results suggest that a combination of physiologic adaptations and improved exercise pain tolerance account for the improvement in walking performance achieved through upper-limb aerobic exercise training in patients with PAD. Furthermore, that both arm- and leg-crank training could be useful exercise training modalities for improving cardiovascular function, walking performance, and exercise pain tolerance in patients with symptomatic PAD. PMID- 16376203 TI - Functional decline in lower-extremity peripheral arterial disease: associations with comorbidity, gender, and race. AB - PURPOSE: To identify comorbidities associated with increased rates of functional decline in persons with lower-extremity peripheral arterial disease (PAD). We also determined whether female sex and black race were associated with greater functional decline than male sex and white race, respectively, in PAD. METHODS: Three-hundred ninety-seven men and women with PAD were followed prospectively for a median of 36 months. The presence of comorbid illnesses was determined with medical record review, patient report, medications, laboratory values, and a primary care physician questionnaire. Functional outcomes, measured annually, included the 6-minute walk, usual-paced and fast-paced 4-meter walking speed, and summary performance score. The summary performance score is a composite measure of lower-extremity functioning (score range, 0 to 12; 12 = best). RESULTS: Adjusting for known and potential confounders, PAD patients with pulmonary disease had a significantly greater average annual decline in 6-minute walk performance of -34.02 ft/y (95% confidence interval [CI], -60.42 to -7.63; P = .012), rapid-paced 4-meter walk speed of -0.028 m/s/y (95% CI, -0.054 to -0.001; P = .042), and summary performance score of -0.460/y (95% CI, -0.762 to -0.157; P = .003) compared with those without pulmonary disease. PAD patients with spinal stenosis had a greater average annual decline in 6-minute walk performance of 77.4 ft/y (95% CI, -18.9 to -35.8; P < .001) and usual-paced 4-meter walking velocity of -0.045 m/s/y (95% CI, -0.081 to -0.009; P = .014) compared with participants without spinal stenosis. CONCLUSION: At 3-year follow-up, pulmonary disease and spinal stenosis were each associated with a significant decline in functioning among persons with PAD. In contrast, female sex and black race were not associated with functional decline among persons with PAD. PMID- 16376204 TI - Mid-term results of endovascular treatment for symptomatic chronic nonmalignant iliocaval venous occlusive disease. AB - BACKGROUND: The goal of this article is to present clinical and patency results of endovascular treatment of nonmalignant, iliocaval venous obstructive disease and to discuss the evolution of technical details. METHODS: From November 1995 to June 2004, 44 patients (female-male ratio, 3.9:1; left-right lower limb ratio, 8.6:1; median age, 42 years; range, 21-80 years) had treatment for chronic disabling obstructive venous insufficiency with iliocaval stenosis or occlusion. The clinical class of CEAP was 2 in 11 limbs, 3 in 31, 4 in 4, 5 in 1, and 6 in 1; etiology was primary in 32 patients, secondary in 10, and congenital in 2. Anatomic involvement included superficial veins in 16 patients and perforator veins in 11. Obstruction was associated with superficial reflux in 4 patients, deep reflux in 13, and both in 13. Ten patients had occlusion. All procedures were performed in the operating room with perioperative angiography and angioplasty with or without self-expanding stent implantation. Venous clinical severity and disability scores were obtained before and after treatment. Patency and restenosis were evaluated by duplex Doppler ultrasonography. RESULTS: No perioperative death or pulmonary embolism occurred. The technical success rate was 95.5% (two recanalization failures), and two (4.5%) perioperative stent migrations occurred. One early thrombosis (2.4%) was treated by thrombectomy and creation of an arteriovenous fistula. One late death and one thrombosis occurred. Restenoses were found in five patients and were all treated successfully (four needed iterative stenting). Median follow-up was 27 months (range, 2-103 months). Median venous clinical severity score improved from 8.5 to 2, and median venous disability score improved from 2 to 0. Cumulative primary, assisted primary, and secondary patency rates of the venous segments at 36 months were 73%, 88%, and 90%, respectively, in intention to treat. The survival rate was 100% at 12 months and 97.3% at 60 months. CONCLUSIONS: Endovascular treatment of benign iliocaval occlusive disease is a safe and efficient minimally invasive technique with good mid-term patency rates. Moreover, it improves cases with obstruction only, as well as cases with associated reflux and obstruction. Primary stenting should always be performed by using self-expanding stents deployed under general anesthesia to avoid lumbar pain. In case of failure, the endovascular procedure does not preclude further surgical reconstruction. PMID- 16376206 TI - Can phlebectomy be deferred in the treatment of varicose veins? AB - OBJECTIVE: This study was designed to observe the clinical sequelae of varicose veins after great saphenous vein (GSV) ablation and to assess possible predictability of spontaneous varicose vein regression. METHODS: Patients with symptomatic varicose veins secondary to GSV insufficiency treated with radiofrequency ablation (RFA) were enrolled in the study. Up to five of the largest varicose veins in each limb were mapped, sized, and documented before RFA. No varicose vein was treated either at the time of RFA or within 6 months postoperatively. Varicose vein status was recorded at follow-up visits. RESULTS: Fifty-four limbs in 45 patients were included. A total of 222 varicose veins were documented before RFA (4.1 +/- 1.1 varicose veins per limb) with an average size of 11.4 +/- 3.7 mm. During the follow-up period, complete resolution of visible varicose veins was seen in 13% of limbs after RFA alone, and 63 (28.4%) varicose veins spontaneously resolved. A further 88.7% (141/159) of varicose veins decreased in size an average of 34.6% (4.3 +/- 3.4 mm). Preoperatively, 19.4% of varicose veins were above the knee and 75.7% were below the knee. Complete varicose vein resolution was 41.9% (18/43) above the knee and 25.6% (43/168) below the knee. For the above-knee varicose veins, 88.4% (38/43) were located medially, and all the resolved ones (47.4%, 18/38) were medial varicose veins. Resolution rates of the 168 below-knee varicose veins were 30.6% (33/108) of medial, 23.1% (6/26) of anterior, 20.0% (3/15) of lateral, and 5.3% (1/19) of posterior. CONCLUSIONS: Great saphenous vein ablation resulted in subsequent resolution or regression of many lower-limb visible varicose veins. With further study, the predictability of varicose vein regression may perhaps be increased, which can then direct the treatment strategy to further leverage the advantages of minimally invasive endovenous procedures. PMID- 16376207 TI - Comparison of elastic versus nonelastic compression in bilateral venous ulcers: a randomized trial. AB - BACKGROUND: There is controversy regarding the effectiveness of elastic vs nonelastic compression in the healing of venous leg ulcers. To gain insight into this controversy, we randomly compared a four-layer elastic bandage with a nonelastic garment (CircAid) in patients with venous leg ulcers. METHODS: Twenty four extremities of 12 patients with bilateral leg ulcers were randomized to have a four-layer elastic bandage in 1 extremity and a nonelastic compression garment CircAid in the contralateral limb. The CEAP classification in 22 extremities was C(6 S); ES; A(D6,14,15,P18); PR; 1 extremity in 2 patients was C(6 S); E(S); A(D 6,14,15,P18); PRO. There were seven men and five women. Age ranged from 45 to 82 years, with a mean of 61 years. All patients had postthrombotic leg ulcers diagnosed clinically by duplex ultrasonography (n = 8) or by phlebography (n = 4). Every 4 weeks, patients had the ulcer area measured in square centimeters by a computerized scanning method and had the limb girth measured in centimeters at the foot, ankle, and calf. The ankle-brachial index was determined in all cases. Color photographs of the ulcer areas were taken monthly to assess healing progress. All patients were compliant. Results were assessed at 12 weeks. A patient satisfaction sheet was filled out by the patient, who circled one of the following scores: 3, very satisfied; 2, moderately satisfied; and 1, not satisfied. Cox proportional hazards models or paired t tests were used for comparison. RESULTS: The 24 limbs were randomized and divided into 2 groups of 12 each. Group A received CircAid, and group B, a four-layer elastic bandage. Duplex scanning showed a pattern of reflux in 11 limbs of each group. One limb in each group had a pattern of obstruction documented by air plethysmography (n = 2). Phlebography demonstrated the anatomic site in both limbs. The initial ulcer area in group A was 48.98 +/- 14.13 cm2 and was 50.08 +/- 18.30 cm2 in group B (P = .9285). The ulcer healing rate was significantly faster in group A compared with group B (hazard ratio, 0.56; 95% confidence interval, 0.33-0.96; P = .0173). Ulcers with hemodynamic obstruction had a protracted course when compared with the contralateral limbs with reflux. There was no significant difference in girth reduction between groups A and B (hazard ratio, 2.36; 95% confidence interval, 0.30-18.52; P = .3580). The ankle-brachial index was normal (> or = 1.0) in all patients. CONCLUSIONS: In compliant patients, venous leg ulcers randomized to nonelastic compression had a significantly faster healing rate per week than ulcers treated by the conventional four-layer compression system. PMID- 16376208 TI - A prospective study of the fate of venous leg perforators after varicose vein surgery. AB - OBJECTIVE: To describe the fate of perforator veins after surgical treatment of varicose veins and factors that influence this. METHODS: This prospective study of 104 patients assessed perforator veins by using duplex ultrasound scanning in 145 limbs before superficial vein surgery for varicose veins. Veins were marked preoperatively with ultrasound guidance and ligated with an open procedure; those missed were later treated with sclerotherapy. Duplex ultrasound scans and air plethysmography were used to confirm surgical success within 1 month and to monitor recurrence at 6 months, 1 year, and 3 years. RESULTS: A total of 850 incompetent perforators were treated, but 5.7% were missed and required further ablation. After 3 years, 75.8% of the limbs had developed further incompetent perforators for a total of 380 incompetent perforators. The number of ultrasound detectable competent perforators had also increased from 356 to 1047 in that time. The incompetent perforators arose by (1) new vessel formation at the site of previous ligation in 152 (40.4%), (2) changes in pre-existing perforator vessels at other sites in 225 (59.2%), and (3) vessels missed at treatment (< 1%). The diameter of the neovascular channels (3.0 +/- 1.0 mm) was greater than the other incompetent perforators (2.7 +/- 1.0 mm; P < .001). The anatomic distribution of the neovascular recurrences was also different, with 63% found in the paratibial region. The number of new incompetent perforators in a limb was associated with the clinical and physiologic severity of venous disease before surgery, but not to body mass index, gender, or age (P < .01). CONCLUSION: This study shows that incompetent perforator recurrence after surgery is far more common than previously recognized and is primarily due to either neovascularization of previously ligated perforators or the development of incompetence in newly detected perforators in association with persistent venous disease rather than due to poor surgery. PMID- 16376209 TI - Clinical and hemodynamic significance of corona phlebectatica in chronic venous disorders. AB - BACKGROUND: Corona phlebectatica (corona) is a clinical sign associated with chronic venous disorders but is not yet included in the CEAP classification. Our aim was to analyze the clinical and hemodynamic correlations of corona to determine its significance and potential usefulness in classification grading and systems such as the CEAP classification. METHODS: A full clinical and duplex ultrasound record of 287 patients was collected by 60 vascular physicians from 10 European countries. They used the same computer software. This was designed to improve the interobserver consistency of computer-assisted procedures for clinical data gathering. Corona was defined as fan-shaped intradermal telangiectases in the medial and sometimes lateral portions of the ankle and foot. This definition was further refined into three severity grades of light, moderate, and severe, according to reference pictures provided by the software. To avoid overpowering the statistical tests, only data from right lower limbs were analyzed. RESULTS: In this series of 287 unselected patients, a corona was detected in 96 right lower limbs (27.7%) and graded as light in 61 (21.1%), moderate in 24 (8.3%), and severe in 11 (3.8%). The presence of a corona and its severity grade were found significantly related to two risk factors of chronic venous disorders (age and personal history of deep vein thrombosis), the presence of so-called venous symptoms, including pain, pruritus, burning sensation, feeling of swelling, cramps, heavy legs; and most individual items of CEAP clinical classes (varicose veins, edema, all skin changes, C5; and open ulcers, C6). Furthermore, among the patients without skin changes (classes C1 to C3), the presence and grade of corona appeared to be significantly associated with the severity of the disease, with increasing CEAP (P < .05), disability (P < .03), and anatomic (P < .01) scores. The presence of a corona and its severity grade were also found significantly related to the CEAP clinical classes themselves (P < .001), the CEAP disability (P < .001) and clinical severity scores (P < .001), and the presence of superficial reflux in the saphenous and nonsaphenous territories (P = .05) and in the perforators (P < .001). The presence of a deep venous reflux was not found to be significant in this series. The relative risk of finding incompetent leg or calf perforators by duplex ultrasound is 4.4 times greater in patients with corona (P < .001). A comparison between the CEAP classification without corona and a modified classification including corona in C3 shows a better reliability coefficient of Cronbach. CONCLUSION: Corona strongly correlates with the clinical severity and hemodynamic disturbances of the disease. The inclusion of corona in the C3 class should probably improve the reliability of the CEAP clinical classes. PMID- 16376210 TI - Computational investigations of a new prosthetic femoral-popliteal bypass graft design. AB - OBJECTIVE: Prosthetic femoral-popliteal bypasses are performed by using an end-to side anastomosis, and disease can develop at the distal end; this can lead to poor long-term patency rates. Disturbed flow characteristics are hypothesized as being a major factor in promoting disease development. The objective of this study was to propose a new prosthetic femoral-popliteal bypass graft configuration specifically engineered to reduce or eliminate certain disease influencing factors that act on the host artery. METHODS: The proposed device contains a streamlined bifurcation toward its distal end that results in two end to-end anastomoses, rather than the single end-to-side anastomosis in the traditional procedure. Comparisons are made between idealized representations of it and the traditional end-to-side anastomosis for specific femoral artery flow rates. Qualitative results in the form of velocity vector plots and wall shear stress contour plots are compared, and quantitative results examine the wall shear stress magnitudes and gradients along the bed and roof of each graft model. RESULTS: Velocity vector plots through each junction suggest that the proposed graft configuration promotes streamlined flow and helps to reduce the magnitude of flow recirculation and separation regions associated with the traditional end to-side anastomosis. At peak velocity, the flow separation region distal to the toe is eliminated, as evidenced by the change in toe wall shear stress from -0.2 Pa in the traditional anastomosis to +0.5 Pa in the proposed device. Normal fully developed flow occurs sooner in the distal host artery, approximately 15 mm downstream from the toe, unlike 20 mm in the conventional case. The proposed design results in reductions of up to 58% in peak wall shear stress and 86% in peak wall shear stress gradient during the decelerative phase of the femoral pulse in the vicinity of the artery bed below the toe. CONCLUSIONS: In vitro tests on the proposed device suggest that the streamlined nature of blood flow through the junction does result in less disturbed hemodynamic conditions within the host artery junction. Abnormal wall shear stress magnitudes and gradients are reduced, and normal fully developed flow occurs sooner in the distal host artery. This suggests that the proposed graft may have design attributes that are relevant in the search for increased prosthetic femoral-popliteal bypass graft patency rates. A drawback of the proposed device is that significant flow recirculation and separation exist within the prosthesis itself. CLINICAL RELEVANCE: The search for a viable alternative to traditional end-to-side anastomosis for prosthetic femoral-popliteal bypass procedures is ongoing. Prosthetic femoral-popliteal bypass procedures have low long-term patency rates, and there is a need for methods that increase the life span of the procedure. Although research is progressing on a variety of different fronts, this study is significant in that it reports on in vitro tests on a potential device that may increase bypass patency. The device is simple, may be manufactured from clinically proven materials, does not require any additional training in its use, and combines attributes of end-to-side anastomoses with those of end-to-end anastomoses. In addition, the design concept behind the device, the natural bifurcation, may be relevant in other aspects of cardiovascular surgery. PMID- 16376211 TI - Effect of intraluminal thrombus on pressure transmission in the abdominal aortic aneurysm. AB - OBJECTIVE: To clarify the effect of intraluminal thrombus on pressure transmission. METHODS: A saccular aneurysm was inserted into an artificial circulation system. Subsequently, the saccular aneurysm was filled with eight different human aortic aneurysm thrombus samples. Starch solution in an empty aneurysm was used as a control. A pressure sensor measured the pressure in the circulation, and a second piezoelectric sensor measured the pressure in the saccular aneurysm at 3, 2, and 1 cm from the endoluminal surface (23 locations). The influence of the elastic characteristics of the aneurysm wall on the extent of pressure reduction was evaluated by experiments performed with aneurysms made of rubber and paraffin. RESULTS: The pressures measured in the empty aneurysm were identical to those measured in circulation (P > .05). The pressure measured in the thrombus was significantly lower than the pressure measured in the circulation (P < .05). The mean pressure ratio between the systolic thrombus pressure and systolic circulation pressure at 1, 2, and 3 cm was 0.90 +/- 0.09, 0.86 +/- 0.10, and 0.81 +/- 0.09, respectively. However, there was a clear correlation between the pressure in the circulation and in the thrombus (Pearson correlation coefficient: mean, r = 0.997; range, 0.975-0.999; P < .01). The change in circulatory pressure was followed by an almost identical change in thrombus pressure (regression coefficient: mean, beta = .997; range, .983-1.000; P < .01). In stiff aneurysms, the pressure reduction is less than in more compliant ones (P < .05). CONCLUSIONS: In an in vitro model, pressure in the aneurysmal sac is reduced by fibrinous thrombus. CLINICAL RELEVANCE: Endovascular aneurysm repair (EVAR) aims at reducing the pressure in the aneurysmal sac. Therefore, it seems attractive to use pressure monitoring in the aneurysmal sac as a follow-up after EVAR. This study contributes to the development of the rationale of pressure monitoring in the aneurysmal sac as a follow-up method after EVAR. The aneurysmal sac is filled with thrombus. To interpret pressure measurements in the thrombus, we have to learn about the effect of the thrombus on pressure transmission and on the pressure measurements themselves. Our study demonstrates that reduction of pressure occurs as it is transmitted through a human aortic thrombus. PMID- 16376212 TI - Vascular endothelial growth factor naked DNA gene transfer enhances thrombus recanalization and resolution. AB - OBJECTIVES: Enhancing thrombus resolution may reduce the long-term complications of venous thrombosis. The aim of this study was to examine whether a sustained release of vascular endothelial growth factor (VEGF) would further improve thrombus recanalization. METHODS: Inferior caval vein thrombosis was induced in a cohort of 21 male Wistar rats. A plasmid encoding the human VEGF gene (phVEGF) was injected directly into thrombus (30 to 50 microg) and the muscle adjacent to the inferior vena cava (300 to 400 microg). A plasmid containing the gene encoding beta-galactosidase (pCMVbeta) was injected into the same sites of a separate cohort of rats to act as a control. Tissues were harvested after 1 and 2 weeks, and beta-galactosidase activity was measured to estimate transfection efficiency. Muscle and serum VEGF were measured by enzyme-inked immunosorbent assay. Thrombus size, recanalization, and organization were determined by computer-assisted image analysis. RESULTS: The efficiency of control plasmid transfection into muscle was about 1%. No serum hVEGF was detected in phVEGF- or pCMVbeta-treated animals. Significantly raised levels of hVEGF (P < .01) were detected in the muscle injected with phVEGF after 2 weeks compared with control muscle. There was a significant reduction in thrombus size of 23% (P < .05) and 48% (P < .001) in phVEGF-treated animals compared with pCMVbeta-treated controls after 1 and 2 weeks, respectively. Thrombus recanalization was a significantly greater in the phVEGF-treated group after 2 weeks (mean 19% +/- 2% [SEM]) compared with controls (mean 13% +/- 2%, P < .01). There were no differences in the thrombus organization score. CONCLUSION: VEGF gene therapy of venous thrombus resulted in smaller thrombi with greater recanalization. Angiogenic gene therapy may form the basis of a novel treatment that may improve the resolution of venous thrombi. CLINICAL RELEVANCE: Deep vein thrombosis may lead to residual venous obstruction or reflux and result in post-thrombotic complications that are debilitating and have a substantial socioeconomic impact. Enhancing the resolution of venous thrombi may reduce post thrombotic complications. PMID- 16376213 TI - Resveratrol inhibits vascular smooth muscle cell proliferation and induces apoptosis. AB - OBJECTIVE: In France, despite a high intake of dietary cholesterol and saturated fat, the cardiovascular death rate is one of the lowest among developed countries. This "French paradox" has been postulated to be related to the high red wine intake in France. The aim of this study was to determine the effects of resveratrol, a major polyphenol component of red wine, on vascular smooth muscle cell (SMC) proliferation in vitro. METHODS: SMCs were exposed to 10(-6) to 10(-4) M resveratrol and cell proliferation was assessed by cell counting. Cell cycle analysis was done by treating cells with propidium iodide followed by flow activated cell sorting. Apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling staining. RESULTS: We demonstrate that resveratrol inhibited bovine aortic SMC proliferation in a dose dependent manner. The lowest concentration of resveratrol resulting in a significant decrease in SMC proliferation compared with control was 10(-5) M. By flow cytometry, we observed a block in the G1-S phase of the SMC cycle. Resveratrol treatment also resulted in a dose-dependent apoptosis of SMCs but had no effects on SMC morphology. CONCLUSION: The results indicated that vascular SMC proliferation could be inhibited by resveratrol through a block on G1-S phase and by an increase in apoptosis. It supports the conjecture that red wine consumption may have a beneficial effect on cardiovascular mortality. CLINICAL RELEVANCE: Our results suggest that resveratrol inhibits, in a dose-dependent manner, smooth muscle cell proliferation, which may help to partially explain a beneficial effect of wine drinking. This inhibition is related to an early block in the cell cycle and also to a dose-dependent apoptotic effect. The present study demonstrates that resveratrol not only is an indirect marker of a healthy life style and alimentation but may also be directly responsible for the French paradox. PMID- 16376214 TI - Role of nitric oxide in resveratrol-induced renal protective effects of ischemic preconditioning. AB - BACKGROUND: Resveratrol, a natural antioxidant and polyphenol found in red wine and grapes, has been found to pharmacologically precondition the heart through upregulation of nitric oxide (NO). This study was designed to explore the involvement of NO in the renoprotective effect of resveratrol in renal ischemic preconditioning in rat kidney. METHODS: Ischemic preconditioning was induced by three cycles 2-minutes of ischemia followed by 5 minutes of reperfusion before 45 minutes of prolonged ischemia. Resveratrol was given 1 hour before the surgical procedures. RESULTS: Ischemic preconditioning and resveratrol treatment significantly improved the renal dysfunction, decrease in total NO levels, and oxidative stress induced by 45 minutes of ischemia followed by 24 hours of reperfusion. Histopatholgic examination of the kidneys of ischemic/reperfusion rats revealed severe renal damage, which was attenuated in both preconditioned and resveratrol-treated animals. Preconditioning and resveratrol administration led to a marked increase in NO levels in kidney. Renoprotective effects of resveratrol were abolished when animals were pretreated with NG-nitro-L-arginine methyl ester, a nonspecific NO synthase inhibitor. CONCLUSIONS: These findings demonstrate an important contributory role of NO in the protection afforded by resveratrol in renal ischemic preconditioning. CLINICAL RELEVANCE: It is now well established that brief periods of ischemia followed by reperfusion render a variety of tissues tolerant to subsequent ischemia/reperfusion-induced injury. This phenomenon, referred to as ischemic preconditioning, was first demonstrated in the dog myocardium. The potential for clinical application of such a powerful protective phenomenon has generated enormous interest in identifying the underlying intracellular signaling pathways, with the ultimate aim of pharmacologically exploiting these mechanisms to develop therapeutic strategies that can enhance tolerance to ischemia/reperfusion injury in patients. This study explored the possible involvement of nitric oxide in renal ischemic preconditioning. PMID- 16376215 TI - Combined open and endovascular treatment of a right subclavian artery aneurysm: a case report. AB - A 69-year-old man with previous ascending aortic repair combined with valve replacement for an ascending aortic aneurysm presented with a type 2 thoracoabdominal aneurysm and a 4.4-cm aneurysm of the right subclavian artery. Because of the anatomic location of the aneurysm and his previous operation, an innominate to carotid artery stent graft and a carotid-subclavian bypass and vertebral artery bypass were performed. Postoperative computed tomographic angiography confirmed good flow in the right carotid and vertebral artery, and the patient recovered without complication. PMID- 16376216 TI - Endovascular repair of symptomatic external carotid artery stenosis. AB - The treatment of external carotid artery stenosis has been described with a variety of operative interventions. We present a patient who presented with amaurosis fugax and a critical left external carotid artery stenosis with known left internal carotid artery occlusion. We treated this stenosis with angioplasty and stenting rather than endarterectomy. Our patient did well and had no complications from the procedure. Endovascular repair of symptomatic external carotid artery stenosis provides an alternative treatment method to conventional endarterectomy for patients with high surgical risk. PMID- 16376217 TI - In situ aortic allograft insertion to repair a primary aortoesophageal fistula due to thoracic aortic aneurysm. AB - Aortoesophageal fistula due to thoracic aortic aneurysm is an uncommon cause of gastrointestinal bleeding and has an extremely poor prognosis. In the English literature, we found only 27 successfully managed cases of primary aortoesophageal fistula due to thoracic aortic aneurysm. We present a case of 74 year-old man who experienced the erosion of a thoracoabdominal aortic aneurysm into the esophagus. We successfully performed resection and replacement of the thoracoabdominal aorta with a cryopreserved allograft and total thoracic esophagectomy. A few months later, the esophagus was reconstructed with orthotopic colonic interposition. The patient recovered well and resumed a normal life (12 months' follow-up). PMID- 16376218 TI - Successful management of concomitant aortoesophageal and aortotracheal fistulae secondary to a thoracic aortic aneurysm: case report and review of literature. AB - Thoracic aortic aneurysm with fistulization into the trachea or the esophagus is usually a lethal complication. An unusual case of combined aortotracheal and aortoesophageal fistulae is presented here. The patient had a successful outcome after resection of the aneurysm with tracheal repair and esophageal resection. A review of literature on this subject is also included in this manuscript. PMID- 16376219 TI - Endograft repair of partially anomalous pulmonary venous connection with dual drainage. AB - A 73-year-old woman with dyspnea and fatigue was found to have a partially anomalous pulmonary venous connection with communication between the right upper lobe pulmonary vein and the superior vena cava, as well as a normal pulmonary vein confluence with the left atrium. She was successfully treated with an endograft implanted in the superior vena cava, covering the defect. The unique dual drainage of the pulmonary vein allowed percutaneous treatment of a condition that ordinarily requires complex caval and intracardiac reconstruction. Except for several episodes of arrhythmias, she has been doing well, with exercise tolerance improving from New York Heart Association class III at baseline to class I after treatment. Follow-up imaging and clinical findings at 36 months showed no migration of the stents and significant improvement of quality of life. PMID- 16376220 TI - Ruptured aneurysm of the inferior thyroid artery: repair with coil embolization. AB - Although pseudoaneurysm of the thyrocervical trunk is seen with increasing frequency as a result of trauma or central venous cannulation, true aneurysm of the thyrocervical trunk is exceedingly rare. A 44-year-old woman presented with acute left neck swelling that progressed rapidly to respiratory distress and subsequent hemothorax. Emergency endotracheal intubation was performed for airway control, and tube thoracostomy was placed for drainage of the pleural space. A review of the literature reveals that this is the second patient known to be successfully treated by arterial embolization of a ruptured aneurysm of the inferior thyroid artery and the fifteenth known case of aneurysm of the inferior thyroid artery. Surgical or endovascular intervention has been successful. Observation has been uniformly fatal. Because life-threatening rupture is possible, the presence of this aneurysm mandates intervention. Endovascular coil embolization is a viable option for treatment of this entity. PMID- 16376221 TI - Transection of the brachial artery after closed posterior elbow dislocation. AB - Although elbow dislocations are common orthopaedic lesions, vascular complications remain rare. We report the cases of three patients who presented with a rupture of the brachial artery after closed posterior dislocation, which is even more uncommon. Arteriograms were performed in all cases because of the persistent absence of pulses at the wrist after emergency reduction. In each patient, the treatment consisted of the insertion of reversed end-to-end saphenous bridges. None of them presented mid-term vascular complications (mean follow-up, 17 months). Brachial artery disruption can result from closed posterior elbow dislocation and responds well to vascular repair. PMID- 16376222 TI - Nontraumatic pseudoaneurysm of the proximal ulnar artery with eosinophilia. AB - Proximal ulnar artery aneurysms, including pseudoaneurysms, have not been described in the English literature. We report a nontraumatic pseudoaneurysm of the proximal ulnar artery with eosinophilia in a 54-year-old man. Radial, coronary, and hepatic artery aneurysms associated with eosinophilia in idiopathic hypereosinophilic syndrome or allergic granulomatous angiitis (Churg Strauss syndrome) have been reported. Although it is unclear in the present case whether eosinophilia was associated with the pseudoaneurysm of the ulnar artery, eosinophil infiltration into the aneurysmal wall may have influenced vascular injury as the cause of the pseudoaneurysm formation. PMID- 16376223 TI - Middle aortic syndrome: surgical treatment in a child with neurofibromatosis. PMID- 16376224 TI - Radiation: basic principles. AB - This communication describes some basic principles of radiation that are relevant for understanding its use in vascular brachytherapy. It describes various characteristics of gamma-rays and beta-particles, including their interaction with matter such as tissues. It also explains different terms and concepts such as radioactivity, half-life, absorbed dose, range which would help the brachytherapy practitioners appreciate the physics aspects of the therapy. PMID- 16376225 TI - Whodunit? Ghost surgery and ethical billing. AB - A senior vascular surgery resident started an autogenous radical-cephalic arteriovenous fistula procedure on a comatose patient in the surgical intensive care unit (SICU), expecting you to arrive momentarily. You were nevertheless unexpectedly detained establishing hemostasis in the main operating suite. You arrived in the SICU as the dressing was being applied. Fistula flows were excellent and there were no operative complications. The resident who began and finally completed the case was highly skilled and in the final month of his vascular training; you had supervised his satisfactory performance of many procedures like this one during the last 2 years. The patient's elderly wife had consented to the procedure, which she was told you would be directly supervising while the resident performed the surgery. When the operation was over you met with her to explain your emergency conflict and assure her that you checked the resident's work and found it entirely satisfactory. She accepted your explanation and was relieved that the operation went well. The patient's multiple comorbidities nevertheless necessitated an extended postoperative stay in the SICU, where you personally cared for him. The resident had dictated routinely that you attended the procedure, and your billing clerk had no reason to doubt the operative report's accuracy when she submitted your surgical fees to Medicare and the patient's private insurer, which paid to their contractual limits without challenge. On many occasions you have had your billings shorted by both. The resident since has graduated. What should you do? PMID- 16376226 TI - Regarding "Cost-effectiveness of different screening strategies for abdominal aortic aneurysm". PMID- 16376228 TI - Arthroscopic versus mini-open rotator cuff repair: a comparison of clinical outcome. AB - PURPOSE: To compare the outcome of patients who underwent rotator cuff repair using all arthroscopic or mini-open repair techniques. TYPE OF STUDY: Retrospective comparative study. METHODS: We retrospectively reviewed 54 patients who underwent either mini-open or arthroscopic rotator cuff repair. Twenty-six patients underwent mini-open repair and 28 patients had arthroscopic repair. Follow-up averaged 33 months (range, 18 to 48 months) for the mini-open group and 19 months (range, 13 to 26 months) for the arthroscopic group. The patient groups were similar with regard to age, activity level, mechanisms of injury, associated findings at surgery, and tear size measured in square centimeters. The outcome for the 2 groups was evaluated using a modified American Shoulder and Elbow Society (ASES) score. Statistical analysis was performed using Pearson correlations and the Student t test. RESULTS: The tear size averaged 2.7 cm2 for the mini-open group and 2.0 cm2 for the arthroscopic group (P = .754). All patients showed significant improvement in their scores for pain, satisfaction, and function at the time of follow-up. The average preoperative and postoperative scores for the mini-open group were as follows: pain 17 and 27 (30 possible points), satisfaction 3 and 9 (10 possible points), function 32 and 53 (60 possible points), and total 52 and 89 (100 possible points) (P < .05). For patients who underwent arthroscopic repair, average preoperative and postoperative scores were as follows: pain 12 and 26, satisfaction 2 and 9, function 28 and 51, and total, 42 and 86 (P < .05). Improvement in scores within each group was significant, but the difference in total scores between the 2 techniques was not statistically significant. CONCLUSIONS: This study confirms that short-term results for arthroscopic and mini-open rotator cuff repair are similar and supports continued use of arthroscopic repair techniques. LEVEL OF EVIDENCE: Level III, retrospective comparative study. PMID- 16376229 TI - Arthroscopic repair of full-thickness rotator cuff tears using bioabsorbable tacks. AB - PURPOSE: The purpose of this study was to evaluate the clinical results of bioabsorbable tacks (Suretac; Smith & Nephew Endoscopy, Mansfield, MA) for the arthroscopic repair of full-thickness rotator cuff tears. TYPE OF STUDY: Prospective case series. METHODS: A prospective clinical review was performed on 53 patients (38 men, 15 women) with full-thickness rotator cuff tears repaired arthroscopically using bioabsorbable tacks. Minimum follow-up was 24 months and patients were evaluated using preoperative and postoperative American Shoulder and Elbow Society (ASES) scores and Short-Form 36 Health Surveys (SF-36). A Constant and Murley score was performed at a minimum of 2 years postoperatively. RESULTS: The average patient age was 51 years (range, 23 to 74 years) and the average time for review was 29 months (range, 24 to 60 months). The average size of rotator cuff tears measured 2.5 cm (range, 1 to 5 cm) and an average of 2 Suretacs (range, 1 to 4) were used for the repair. Ten patients had tears larger than 3 cm and 5 patients had tears that measured 5 cm or larger. The average total score according to the ASES shoulder index improved from 33 to 85 points. The average score for pain improved from 6.9 to 1.3 points. The score for function improved from 11 to 25 points. These results were statistically significant (P < 0.01). There was a significant improvement in all components of the SF-36 survey. The average postoperative Constant score was 87 with the pain component measuring 12 points, range of movement 38 points, and power 21 points. CONCLUSIONS: Arthroscopic repair of full-thickness rotator cuff tears using bioabsorbable tacks produces satisfactory outcomes with regard to objective orthopaedic criteria as well as overall patient satisfaction in terms of pain relief and function. Arthroscopic rotator cuff repairs are technically demanding procedures. The use of tacks facilitates this procedure without any apparent deterioration in clinical results. LEVEL OF EVIDENCE: Level IV, case series. PMID- 16376230 TI - Use of preoperative magnetic resonance imaging to predict rotator cuff tear pattern and method of repair. AB - PURPOSE: To determine the magnetic resonance imaging (MRI) criteria for predicting rotator cuff tear pattern and method of repair. TYPE OF STUDY: Retrospective MRI/arthroscopy correlation. METHODS: Sixty-six preoperative MRI scans were evaluated. The maximum medial to lateral length (L) of the tear was measured on T2-weighted coronal cuts. The maximum anterior to posterior width (W) was measured on T2-weighted sagittal cuts. The cases were divided into 3 groups: group 1, short-wide tears, L < or = W, L < 2 cm; group 2, long-narrow tears, L > W, W < 2 cm; and group 3, long-wide tears, L > or = 2 cm, W > or = 2 cm. RESULTS: Of the 66 MRI scans, 55 were adequate for standardized measurement. Group 1, 16 cases: 15 were found at arthroscopy to be crescent-shaped tears repaired end-to bone; 1 was repaired with interval slides. Group 2, 22 cases: all 22 were repaired side-to-side/margin convergence. Group 3, 17 cases: 12 required interval slides, 1 partial repair was performed, and 4 were repaired side-to-side/margin convergence. CONCLUSIONS: Tear pattern and method of repair can be predicted on high-quality MRI scan. Group 1, L < or = W and L < 2 cm, predicts a crescent shaped tear and end-to-bone repair (positive predictive value, 93.8%). Group 2, L > W and W < 2 cm, predicts a longitudinal tear and side-to-side/margin convergence repair (positive predictive value 100%). Group 3, L > or = 2 cm and W > or = 2 cm, predicts a massive contracted tear and that primary end-to-bone or side-to-side repairs are usually not possible and that interval slides or partial repair may be necessary (positive predictive value, 76.5%). The overall diagnostic model based on usable MRI scans significantly predicted arthroscopic findings (P < .001 for chi-square test). LEVEL OF EVIDENCE: Level III, development of diagnostic criteria with universally applied reference (nonconsecutive patients). PMID- 16376231 TI - Arthroscopic double-bundle augmentation of posterior cruciate ligament using split Achilles allograft. AB - PURPOSE: The purpose of this study was to assess the outcome of arthroscopic double-bundle posterior cruciate ligament (PCL) augmentation using split Achilles allograft. TYPE OF STUDY: Prospective case series. METHODS: We analyzed 27 knees in 26 patients whose PCL had been augmented by the arthroscopic double-bundle technique using split Achilles allograft. There were 19 male and 7 female patients with a mean age of 27.9 years. Follow-up averaged 25 months (range, 12 to 48 months). The clinical results were evaluated according to the Lysholm knee scores. The posterior laxity was radiographically measured with differences of posterior tibial translation between the injured and the uninjured knee. RESULTS: The Lysholm knee scores improved from 59.5 to 91.8 points (P < .05). The average radiographic side-to-side difference of the posterior tibial translation was 12.7 mm (range, 10 to 26 mm) preoperatively and 2.4 mm (range, 0 to 8 mm) at the time of the latest follow-up (P < .05). Eighteen knees (67%) had 0 to 2 mm translation, 6 knees (22%) had 3 to 5 mm, and 3 knees (11%) had 6 to 10 mm. CONCLUSIONS: PCL injuries can be successfully treated with arthroscopic double bundle augmentation using split Achilles allograft. This technique can preserve remnant fibers and restore both anterolateral and posteromedial bundles of the PCL. LEVEL OF EVIDENCE: Level IV, case series. PMID- 16376232 TI - A biomechanical comparison of Bosworth and poly-L lactic acid bioabsorbable screws for treatment of acromioclavicular separations. AB - PURPOSE: The purpose of this study was to determine the failure load of intact coracoclavicular ligaments and the pullout strength of a poly-L lactic acid screw (PLLA, Linvatec, Largo, FL) compared with a titanium screw (Bosworth, Stryker Howmedica Osteonics, Allendale, NJ) in the repair of fifth-degree acromioclavicular separations. TYPE OF STUDY: Biomechanical cadaveric study. METHODS: The ultimate tensile strength of coracoclavicular ligaments was determined using a servohydraulic testing machine (MTS, Eden Prairie, MN) by creating grade V acromioclavicular separations. Titanium Bosworth screws and Linvatec PLLA screws were placed across the reduced coracoclavicular joint and through the base of the coracoid process of right and left matched cadaveric shoulder specimens, respectively. The axial pullout strength was determined by MTS uniaxial tensile testing along the screw axis to determine the ultimate fixation failure load at a rate of 1 mm/second. RESULTS: Intact coracoclavicular ligaments required an axial load of 340.15 +/- 100.77 N to fail. The bioabsorbable PLLA screw provided fixation strength that was significantly less than the Bosworth screw fixation strength: 272.01 +/- 63.26 and 367.01 +/- 111.54 N, respectively (P < .05, paired t test). CONCLUSIONS: The titanium Bosworth screw restored strength to the acromioclavicular joint equivalent to the intact coracoclavicular ligament; however, the bioabsorbable PLLA screw fixation was significantly weaker than the intact ligaments and the Bosworth repair. CLINICAL RELEVANCE: Titanium Bosworth screws supply more strength of fixation in grade V acromioclavicular separations than the PLLA screw tested in this study. PMID- 16376233 TI - Suture strength and angle of load application in a suture anchor eyelet. AB - PURPOSE: To assess the effect of suture material, anchor orientation, and anchor eyelet design on the static loading properties of suture anchors. TYPE OF STUDY: Biomechanical bench study. METHODS: Two metallic suture anchors, Mitek GII (Mitek, Westwood, MA) and Corkscrew (Arthrex, Naples, FL) and a bioabsorbable anchor (Biocorkscrew; Arthrex) were tested with single strand of No. 2 Ethibond (Ethicon, Norderstedt, Germany) or No. 2 FiberWire (Arthrex) suture. Suture pull angle was varied through 0 degrees, 45 degrees, and 90 degrees with the anchor rotation angle in either a sagittal or coronal plane. Constructs were tested to failure using an MTS 858 Bionix testing machine (Material Testing Systems, Eden Prairie, MN). Peak loads, stiffness, energy to peak load, and failure modes were determined for all samples. RESULTS: FiberWire showed superior static mechanical properties when compared with single-strand Ethibond over all testing conditions (P < .05). Suture pull angle had a significant effect on load to failure with both metallic anchors but not on the bioabsorbable anchor (P < .05). CONCLUSIONS: Suture pull angle and anchor rotation angle play an important role in the failure load of suture when placed in an eyelet. The polyaxial nature of the Biocorkscrew eyelet allows for increased degrees of freedom but introduces failure of the suture eyelet as a new failure mode. CLINICAL RELEVANCE: The loading direction and placement of the suture anchor plays a role in the performance of the suture anchor-suture complex. PMID- 16376234 TI - Compressive loads in longitudinal lateral meniscus tears: a biomechanical study in porcine knees. AB - PURPOSE: To determine the displacement forces across a lateral meniscal tear during motion. TYPE OF STUDY: Experimental laboratory biomechanical study. METHODS: A middle third longitudinal lateral meniscal cut was created arthroscopically at the "red-white" junction in 5 intact porcine knees. With a pressure transducer in the tear, the knees were repeatedly cycled through a full range of motion. Pressure data were gathered with the knees held at neutral, internal rotation (IRot), and external rotation (ERot) and matched to the specific flexion angle measured by electronic goniometer. Averaged pressure measurements were calculated at each 5 degrees interval. RESULTS: The highest pressures were seen at full extension (neutral, 589 mm Hg; IRot, 1,110 mm Hg; ERot, 337 mm Hg) and declined to a low at 90 degrees of flexion (neutral, 133 mm Hg; IRot, 314 mm Hg; ERot, 187 mm Hg). Then the pressures increased steadily after 100 degrees as the knees were further flexed. The highest pressure was always seen with IRot. IRot during flexion resulted in higher lateral meniscus compressive loads than ERot. CONCLUSIONS: This model demonstrated that a middle third longitudinal lateral meniscal cut is compressed throughout the full range of knee motion. At no time were negative intrameniscal tear pressures registered that would suggest meniscal cut separation. CLINICAL RELEVANCE: These data suggest that meniscal compressive loads, not distractive loads, occur throughout knee flexion and extension. The absence of distractive loads across a meniscal cut suggests that the ability of a repair to align the meniscal fragment may be more important than a high load to failure strength. PMID- 16376235 TI - Localized synovial hypertrophy in the anteromedial compartment of the osteoarthritic knee. AB - PURPOSE: The purpose of this study was to investigate the clinical and arthroscopic findings of osteoarthritis (OA) of the knee with localized synovial hypertrophy in the anteromedial compartment and the clinical results of arthroscopic partial synovectomy. TYPE OF STUDY: Retrospective case series. METHODS: We treated 19 osteoarthritic knees with localized synovial hypertrophy in 19 patients by arthroscopic partial synovectomy under local anesthesia. All patients had complained of knee pain (mechanical type pain) and catching sensations preoperatively. Preoperative diagnosis based on physical findings and imaging studies was a medial meniscus tear with medial OA in all patients. The mean follow-up was 37 months (range, 28 to 46 months). Clinical results were assessed with the Hospital for Special Surgery (HSS) scoring scale, overall subjective estimation, and patient satisfaction. RESULTS: The catching sensation resolved immediately after surgery in all patients. The mean HSS score improved from 56.8 to 72.4 points (P < .01). Nine patients (47.4%) were rated excellent or good according to the overall subjective estimation, and 11 patients (57.9%) were satisfied with the treatment. Both HSS score and patient satisfaction were higher in patients (10 patients) who complained of intraoperative pain during synovectomy than in those (9 patients) who hardly felt the pain. CONCLUSIONS: Localized synovial hypertrophy in the anteromedial compartment of OA knees occasionally caused symptoms of pain and catching sensations that resembled meniscal symptoms. Arthroscopic partial synovectomy was effective, especially for those who complained of intraoperative pain during synovectomy under local anesthesia. LEVEL OF EVIDENCE: Level IV. PMID- 16376236 TI - Arthroscopic assessment of cartilage repair: a validation study of 2 scoring systems. AB - PURPOSE: This study tested the validity and reliability of the International Cartilage Repair Society (ICRS) cartilage repair assessment and the Oswestry Arthroscopy Score (OAS), which have been designed to assess repair of articular cartilage. TYPE OF STUDY: Prospective validation study of arthroscopic cartilage repair scores. METHODS: Arthroscopic videos were assessed by a panel of orthopaedic surgeons specializing in cartilage repair. Scoring was repeated after a 2-month interval. Scorers also answered a questionnaire to assess the face and content validity of the scoring systems. Validity of the 2 systems was compared and reliability and repeatability were measured. Pearson's correlation coefficient was used to measure equivalence reliability. The interclass correlation coefficient (ICC) was used to assess the repeatability and inter rater reliability of each score, and internal consistency was assessed with Cronbach's alpha. RESULTS: Face and content validity are acceptable for both scores. There is good agreement (equivalence reliability) between the scores (Pearson's correlation coefficient, r = .88; P < .001). Stability (interobserver reliability) and repeatability (test-retest reliability) are satisfactory for both scores with an ICC >0.7 for each score. Cronbach's alpha was 0.91 for ICRS and 0.82 for OAS, indicating better internal consistency for the ICRS score. CONCLUSIONS: The ICRS and OAS arthroscopic scores have been validated for the assessment of cartilage repair and both have been found to be statistically reliable and repeatable. The ICRS score does not allow for graft hypertrophy and may overscore in this situation, whereas the OAS includes assessment of graft stiffness. Both scores show satisfactory stability and repeatability. Internal consistency is adequate for both scores, although it is higher for the ICRS score. Both the ICRS and OAS arthroscopic scores are effective tools in the evaluation of cartilage repair. LEVEL OF EVIDENCE: Level III, diagnostic study of nonconsecutive patients (no consistently applied reference gold standard). PMID- 16376237 TI - Evaluation of the neurosensory function of the medial meniscus in humans. AB - PURPOSE: Menisci are known to have receptors mainly concentrated at the anterior and posterior horns. Although they are purported to send afferent impulses to the central nervous system, this function has not been thoroughly evaluated. The purpose of the study was to investigate whether stimulation of the menisci initiates a cortical response. The reaction of the end organ to the reflex arc is also evaluated. TYPE OF STUDY: Prospective case series. METHODS: Fourteen patients with normal medial menisci were included in the study. Different parts of the knee joint (the posterior horn and the body of the medial meniscus, the medial femoral condyle, the capsule, and the joint space) were electrically stimulated by a probe during arthroscopy. The cortical response was monitored with somatosensory-evoked potentials (SEPs). The compound muscle action potentials (CMAPs) of the semimembranosus, quadriceps, and biceps femoris muscles were also monitored with electroneuromyography (ENMG). RESULTS: Among the stimulated parts, only the posterior horn of the meniscus produced cortical responses. No response was obtained with stimulation of the medial femoral condyle, the body of the medial meniscus, the capsule, or the joint space. Stimulation of the posterior horn of the medial meniscus produced a measurable amount of CMAP latency for the semimembranosus muscle, but not for the quadriceps and biceps femoris muscles. CONCLUSIONS: Stimulation of the posterior horn of the medial meniscus produces reproducible cortical SEPs and results in ENMG-verified response of the semimembranosus muscle where no response of the semimembranosus muscle is detected with stimulation of the other parts of the knee. CLINICAL RELEVANCE: The knowledge that only the horns of the medial meniscus have mechanoreceptors in the medial compartment of the knee helps to understand patients' signs and symptoms in medial compartment disease. PMID- 16376238 TI - Percutaneous correction (humeroplasty) of humeral head defects (Hill-Sachs) associated with anterior shoulder instability: a cadaveric study. AB - PURPOSE: The goal of our study was to determine whether a bone tamp could be used to correct a Hill-Sachs lesion by a unique technique that we developed termed "humeroplasty." TYPE OF STUDY: In vitro cadaveric study. METHODS: We created reproducible Hill-Sachs lesions in 14 cadaveric humerii, which we then reduced with a curved bone tamp using our proposed technique. Paired t tests, with the significance set at P < .05, were then used to compare the prereduction and postreduction depths, widths, lengths, and volumes of the lesions. RESULTS: The average prereduction defect measurements were 8.9 mm deep, 16.3 mm wide, 24.1 mm long, and 1,755 mm3 in volume. The average postreduction defect measurements were 1.6 mm deep, 6.2 mm wide, 10.4 mm long, and 50.3 mm3 in volume. A statistically significant improvement (P < .001) was seen in all parameters. CONCLUSIONS: Our technique was able to consistently and significantly restore all measured parameters of the Hill-Sachs lesions using a curved bone tamp. This reduction to a "small" defect may reduce the higher rate of recurrence normally found with the surgical treatment of shoulder instability when "large" Hill-Sachs lesions are present. CLINICAL RELEVANCE: Humeroplasty potentially might reduce the higher rate of recurrence observed when the surgical treatment of shoulder instability involves the presence of large Hill-Sachs lesions. PMID- 16376239 TI - Radiofrequency energy effects on the mechanical properties of tendon and capsule. AB - PURPOSE: To compare the mechanical properties of tendon and capsule after radiofrequency (RF) energy treatment. TYPE OF STUDY: An in vitro study. METHODS: RF energy was applied to ovine extensor tendon and human cadaveric glenohumeral capsule varying in the treatment wattage and time (5, 10, or 20 W for 10 or 30 seconds). The associated tissue length changes and dynamic and failure properties of the tissues were investigated using a materials testing machine. RESULTS: Length changes in the 2 tissues were comparable across the range of treatment settings used with both increases in the treatment wattage and time increasing the amount of tissue shrinkage observed. However, tendon showed greater changes in its mechanical properties after RF treatment, with significant decreases in the failure properties of the tissue as well as the dynamic and static stiffness. CONCLUSIONS: RF treatment shrinks collagenous tissues in a progressive manner correlated to the treatment wattage. However, it has different effects on the mechanical properties of tendon and capsule with the properties of tendinous tissues dramatically reduced. CLINICAL RELEVANCE: RF treatment has been shown to effect the mechanical properties of different collagenous tissues differently; therefore, it must be used specifically and with caution around areas of mixed tissue origin. PMID- 16376240 TI - Arthroscopic dorsal radiocarpal ligament repair. AB - PURPOSE: The goal of this study was to determine if arthroscopic repair of a dorsal radiocarpal ligament (DRCL) tear is effective in ameliorating wrist pain. TYPE OF STUDY: Retrospective study. METHODS: A chart review was performed of 53 patients (56 wrists) who underwent wrist arthroscopy with use of a volar radial portal. There were 21 patients with DRCL tears. Mean follow-up was 16 months. Thirteen patients underwent arthroscopic DRCL repair and/or thermal shrinkage (5 repairs, 6 repair plus shrinkage, and 2 shrinkage). Lunotriquetral tears were treated with debridement and pinning. Triangular fibrocartilage (TFC) tears were debrided or repaired. Scapholunate ligament tears/instability were treated with capsulodesis. RESULTS: The 4 patients who underwent repair of an isolated DRCL tear had excellent results with no or mild pain. All returned to their previous occupation. Dorsal capsulodesis was performed in 7 patients with 4 fair/poor results. Nine DRCL repairs/shrinkage were in association with other procedures for ulnar-sided pathology with 6 fair/poor results. CONCLUSIONS: Tears of the DRCL are more common than previously suspected. They are best seen through a volar radial portal and are amenable to arthroscopic repair. DRCL tears appear to be part of a spectrum of radial and ulnar-sided carpal instability as evidenced by the frequent association with scapholunate ligament tears/instability or ulnar sided pathology. Isolated DRCL tears can be solely responsible for wrist pain. Good results are obtained with arthroscopic repair of isolated DRCL tears. The presence of a DRCL tear when seen in combination with a scapholunate, lunotriquetral, or TFC tear connotes a greater degree and/or duration of carpal instability, and portends a poorer prognosis following treatment. Recognition of this condition and further research into treatment methods is needed. LEVEL OF EVIDENCE: Level IV. PMID- 16376241 TI - Arthroscopic debridement of the elbow for arthrofibrosis resulting from nondisplaced fracture of the radial head. AB - PURPOSE: Mason type I fractures of the radial head consist of fractures that occur without displacement. Arthrofibrosis is a rarely reported complication of Mason I radial head fractures. Symptoms include pain, stiffness, and crepitus. We assessed the efficacy of an arthroscopic procedure including debridement and capsular release for patients with persistent symptoms and failure of nonoperative therapy. TYPE OF STUDY: Therapeutic study, case series. METHODS: Retrospective analysis of functional outcome data was carried out for all eligible cases treated at our institution between 1995 and 2003. Twenty procedures were performed, with 8 patients lost to follow-up. The outcome data consisted of range of motion measurements and functional indices derived from the Mayo Performance Index (MPI). Mean follow-up duration was 54 months. RESULTS: Surgical findings included extensive scarring in the radiocapitellar joint, cartilage loss in the radial head and capitellum, scarring and synovitis in the ulnohumeral joint, and scarring with adhesions in the posterior compartment and posterolateral gutter. The mean MPI score preoperatively was 64.1 and the mean postoperative score was 89.5. Total range of motion arc rose from 108 degrees preoperatively to 126 degrees postoperatively. CONCLUSIONS: The outcome data suggest that arthroscopic debridement and capsular release is an effective method of treating arthrofibrosis resulting from Mason I radial head fractures. LEVEL OF EVIDENCE: Level IV. PMID- 16376242 TI - Arthroscopic labral repair in the hip: surgical technique and review of the literature. AB - Hip pain can be caused by multiple pathologies. Injuries to the acetabular labrum are the most common pathologic findings identified at the time of hip arthroscopy. Five causes of labral tears have been identified; these include trauma, femoroacetabular impingement, capsular laxity, dysplasia, and degeneration. Studies have shown the function and the importance of the labrum. To restore function of the labrum, new surgical techniques, such as suture anchor repair, have been described. The goal of arthroscopic treatment of a torn labrum is to relieve pain by eliminating the unstable flap tear that causes hip discomfort. The goals of these treatments are to maintain the function of the hip joint and decrease the development of premature arthrosis. PMID- 16376243 TI - Rapid progression of chondral disease in the lateral compartment of the knee following meniscectomy. AB - We present 2 cases of severe, rapidly progressive chondral disease in the lateral compartment within 12 months after meniscectomy. In both cases, the lateral compartment was salvaged with simultaneously performed cartilage repair techniques and meniscal transplantation. The first case is of a 16-year-old boy who suffered a complex irreparable posterior horn lateral meniscus tear that was treated with an aggressive partial meniscectomy, and developed a rapid onset of severe lateral compartment symptoms associated with a focal grade IV chondral defect of the lateral femoral condyle within 10 months of his index meniscectomy. The second case is that of an athletic 43-year-old orthopaedic surgeon who suffered a complex lateral meniscus tear that required a near total lateral meniscectomy. Within 5 months of the lateral meniscectomy, he developed severe lateral symptoms with a focal grade IV chondral defect of the lateral femoral condyle. In both cases, the articular cartilage defects were treated with osteochondral grafting at the time of lateral meniscus transplantation with excellent results at 2-year follow-up. These cases highlight the significant need for a heightened the awareness of the relatively increased risk of rapid lateral compartment degeneration following lateral meniscectomy. This, in combination with the appropriate use of cartilage restoration techniques, provides the potential to salvage or prevent rapid onset, unicompartmental degenerative disease, and the ability to reduce symptoms and improve function in these challenging patients. PMID- 16376244 TI - Safe arthroscopic access to the central compartment of the hip. AB - This technical note describes a new method that allows access to the central compartment of the hip under arthroscopic control via the peripheral compartment with less risk of labral perforation and/or cartilage scuffing. After placement of the anterolateral portal in the peripheral compartment without traction, the anterior head area with the anterior acetabular labrum and the anterior surface of the femoral head are inspected. Under arthroscopic control, a guidewire is introduced through the anterior portal in between the anterior labrum and anterior femoral head cartilage and into the central compartment. The arthroscope is then removed from the anterolateral portal, the hip distracted, and the arthroscope introduced via cannulated instruments over the guidewire into the central compartment. Further portal placement can be controlled arthroscopically. PMID- 16376245 TI - Arthroscopic excision of the symptomatic discoid medial meniscus in one piece: a surgical technique. AB - We introduce a surgical technique of arthroscopic excision of the symptomatic discoid medial meniscus in one piece. Compared with other surgical techniques of partial meniscectomy, our technique has several advantages. It is easier, less aggressive, there is less formation of foreign bodies, and it is a time-saving technique. PMID- 16376246 TI - Arthroscopic Lapidus arthrodesis. AB - Lapidus arthrodesis is the surgical treatment of choice for symptomatic hypermobility of the medial cuneiform metatarsal joint that is not responding to conservative treatment. The open procedure was criticized for its prolonged healing, high nonunion rate, and the tendency for dorsal angulation of the first metatarsal. We describe an arthroscopic approach that has the advantage of more thorough preparation of the fusion site with minimal bone removal and better control of the arthrodesis position and with less chance of malunion. PMID- 16376247 TI - Arthroscopic glenoid resurfacing with meniscal allograft: a minimally invasive alternative for treating glenohumeral arthritis. AB - The chronically painful arthritic glenohumeral joint recalcitrant to nonsurgical treatment modalities generally has been treated with an open arthroplasty type of procedure. Certain patients may benefit from a less invasive surgical technique in which a meniscal allograft is used to resurface the glenoid, resulting in decreased pain and increased function. We describe an arthroscopic method of glenoid resurfacing with a meniscal allograft to aid in the restoration of function by providing pain relief to patients debilitated by arthritic conditions of the glenohumeral joint. PMID- 16376248 TI - Perioperative morbidity and 30-day mortality after intertrochanteric hip fractures treated by internal fixation or arthroplasty. AB - The conventional treatment for intertrochanteric hip fracture is open reduction and internal fixation. However, hip arthroplasty is occasionally performed. The objective of this study was to determine the 30-day mortality for patients with intertrochanteric hip fracture treated with open reduction, internal fixation, or hip arthroplasty. The mortality rate for patients treated with arthroplasty at 4.8% (23/478) was slightly, but not significantly, higher than that for patients treated with open reduction and internal fixation at 4.5% (62/1395). However, more of the patients in the arthroplasty group exhibited serious intraoperative cardiorespiratory disturbances (62% vs 22%) and died in the hospital (77% vs 35%) when compared with the patients in the open reduction and internal fixation group. Although the incidence of 30-day mortality in these groups was not significantly different, the patients in the arthroplasty group were more likely to have a complicated intraoperative course and die in the hospital. PMID- 16376249 TI - Blood loss after total hip replacement: a prospective randomized study between wound compression and drainage. AB - A randomized, controlled study compared the effects of wound compression with drainage after primary total hip arthroplasty. In 51 patients, an inflatable cuff was placed over the wound underneath a girdle (System Calmed, Calmed AB, Askim, Sweden). Control patients had wound drainage (n = 54). Preoperative and intraoperative variables did not differ between groups. Total blood loss was calculated using hemoglobin balance; with compression it was 1510 +/- 656 mL (mean +/- SD) and in controls 1695 +/- 712 mL (P = .13). However, less blood was transfused in the compression group (P = .05). Wound infection was seen in 2 patients with compression and in 3 controls. Deep venous thrombosis occurred in 3 controls. Wound discharge was more frequent in controls (19/54 vs 8/51; P = .04). Thus, wound compression had no obvious negative effects and reduced wound discharge and need for transfusion. It may replace drainage after total hip arthroplasty. PMID- 16376250 TI - Perioperative blood loss in cementless or hybrid total knee arthroplasty without patellar resurfacing: a prospective, randomized study. AB - The use of cement is considered as an important way to control perioperative blood loss in knee arthroplasty. We prospectively randomized 57 patients (60 knees) who underwent total knee arthroplasty with (30 knees) or without (30 knees) tibial cement to evaluate perioperative blood loss. The measured total blood loss did not differ significantly between the 2 groups (with tibial cement, 731 +/- 288 mL; without cement, 731 +/- 331 mL; P = .9117). The red blood cell count, hemoglobin level, and hematocrit returned to the preoperative levels within 3 months in both groups. Therefore, tibial cement does not appear to affect perioperative blood loss. This finding has implications when planning blood replacement in cementless and hybrid-type arthroplasties. PMID- 16376251 TI - Effect of the timing of tourniquet release on perioperative blood loss associated with cementless total knee arthroplasty: a prospective randomized study. AB - The control of perioperative blood loss is a major concern in cementless knee arthroplasty surgery. We randomized retrospectively 55 patients (60 knees in total) who had undergone cementless total knee arthroplasty and determined the efficacy of tourniquet release either "before" (30 knees) or "after" (30 knees) wound closure. Measurements of total blood loss showed significant differences between the groups ("Before" group, 906 +/- 238 mL; "After" group, 731 +/- 332 mL; P = .0225). The levels of red blood cells, hemoglobin, and hematocrit recovered to the preoperative levels by 3 months after surgery in both groups. In summary, we recommend that the tourniquet be released after wound closure and that a compressive dressing be applied with the aim of limiting or reducing perioperative blood loss without significant effects of the transfer requirement in the 2 groups, provided that tourniquet time is kept below 60 minutes to avoid ill effects of the tourniquet. PMID- 16376252 TI - Early healing with locked condylar plating of periprosthetic fractures around the knee. AB - Management of periprosthetic fractures around the knee is often difficult because of poor bone quality, comminution, and constraints imposed by the existing prosthesis. Locked condylar plates may provide more reliable fixation for these fractures than traditional methods of fixation. Eleven patients with periprosthetic fractures around the knee were treated with open reduction and internal fixation using a locked condylar plate. All 9 acute fractures and 1 of 2 periprosthetic nonunions healed at an average of 21 weeks, and no patient required additional surgery. Nine fractures healed in anatomical alignment, whereas 1 healed in 5 degrees valgus. The remaining periprosthetic nonunion developed a persistent nonunion with subsequent hardware failure. The average range of motion was 4 degrees to 92 degrees . Locked plating systems are highly effective for the management of complex periprosthetic fractures around the knee. They result in reliable fracture healing and permit early motion in complex fractures. PMID- 16376253 TI - Waiting for hip arthroplasty: economic costs and health outcomes. AB - This prospective cohort study of 153 patients aimed to determine the economic and health costs of waiting for total hip arthroplasty (THA). Health-related quality of life, using self-completed WOMAC and EQ-5D questionnaires, was assessed monthly from enrolment preoperatively to 6 months postsurgery. Monthly cost diaries were used to record costs. The mean waiting time was 5.1 months and mean total cost of waiting for surgery was NZ 4305 dollars(US 2876 dollars) per person (pp) (NZ 1 dollar = US 0.668 dollar). Waiting more than 6 months was associated with a higher total mean cost (NZ 4278 dollars/US 2858 dollars pp) than waiting less than 6 months (NZ 2828 dollars/US 1889 dollars pp; P < .01). Improvements from preoperative to postoperative WOMAC and EQ-5D scores were identified (P < or = .01). Waiting longer led to poorer physical function preoperatively (P < or = .01). Those with poor initial health status showed greater improvement in WOMAC (P = .0001) and EQ-5D (P = .003) measures by 6 months after surgery. Longer waits for total hip arthroplasty incur greater economic costs and deterioration in physical function while waiting. PMID- 16376254 TI - Isolated revision of failed metal-backed patellar components: outcome with minimum 4-year follow-up. AB - Several reports document high failure rates of metal-backed patellar components, but few report the outcome of revising these components to all-polyethylene, cemented implants. At a mean 87.2-month follow-up, we describe a series of 36 patients (40 knees) who underwent isolated metal-backed patellar revision to a cemented, all-polyethylene patellar component. After the patellar revision, 5 patients (6 knees) underwent additional surgery, but no patellar components required revision. The additional surgeries were performed at an average of 77.6 months after patellar revision and included 3 tibial insert exchanges for polyethylene wear, 2 revisions of femoral and tibial components for osteolysis, and 1 realignment procedure for recurrent subluxation of the patella. We conclude that revision of a failed metal-backed patellar component to a cemented, all polyethylene patella is a durable, successful procedure. PMID- 16376255 TI - Evaluation of a porous tantalum uncemented acetabular cup in revision total hip arthroplasty: clinical and radiological results of 60 hips. AB - Long-term survival of the acetabular component in revision total hip arthroplasty (THA) is determined by initial implant stability and bone ingrowth. Highly porous Trabecular Metal (TM, Zimmer, Inc, Warsaw, Ind) facilitates ingrowth and may be an ideal material for revision THA. The outcome and survivorship of a TM, nonmodular, uncemented acetabular cup were assessed in 60 consecutive patients undergoing revision THA (mean follow-up, 42 months; range, 14-68 months). Most (55/60) acetabular cups were implanted without screw fixation. Mean Harris Hips Scores improved from 74.8 preoperatively to 94.4 at last follow-up. Serial radiographs demonstrated excellent bone apposition and bone graft incorporation. Complications were limited to 7 cases of dislocation and 1 revision for aseptic loosening. The TM acetabular cup appears suitable for use in revision THA and warrants further study. PMID- 16376256 TI - Clinical results in valgus total knee arthroplasty with the "pie crust" technique of lateral soft tissue releases. AB - Numerous methods for creating symmetric flexion and extension gaps during knee arthroplasty in valgus knees have been proposed, and no consensus exists about the optimal technique. The "pie crust" technique for lateral soft tissue releases has been used extensively, yet few clinical results have been published. In this study, the clinical outcomes of 24 consecutive knees in 24 patients in whom this method was used in conjunction with a cemented posterior-stabilized prosthesis were evaluated. At a mean of 54 months' (range 24-69 months) follow-up, the knees were performing well with a mean Knee Society score of 97 (range 87-100) and mean range of motion of 121 degrees (range 100 degrees -145 degrees). Importantly, there were no clinical failures or cases of postoperative instability and no cases of radiographic loosening or wear. PMID- 16376257 TI - Intraoperative frozen section for detecting active infection in failed hip and knee arthroplasties. AB - The authors reported the result of 40 sessions of intraoperative frozen section on polymorph count in 33 patients undergoing exploration or revision hip and knee surgery to detect active infection. The correlation rate between frozen section and permanent section was 0.95 to 1.00. If the polymorph count more than 5 per high-power field (40 X) was chosen as positive, the sensitivity, specificity, positive predictive value, and negative predictive value would be 0.93, 0.77, 0.68, and 0.95, respectively. Taking polymorph count of more than 10 as positive, the sensitivity, specificity, positive predictive value, and negative predictive value would then be 0.86, 0.85, 0.75, and 0.92, respectively. The authors concluded that intraoperative frozen section is an inexpensive, rapid, and helpful adjunct to detect active infection. A polymorph count of lower than 5 highly suggests the absence of active infection. PMID- 16376258 TI - Total hip arthroplasty in developmental high dislocation of the hip. AB - In total hip arthroplasty for developmental high dislocations, placement of the implant cup in the true acetabulum and femoral-shortening osteotomy can produce satisfactory results. We performed total hip arthroplasties in 25 high dislocated hips (22 patients) between 1992 and 2000, placing all cups in the true acetabula and using noncemented components and performing a femoral-shortening osteotomy in 22 hips. The overall complication rate was 36%. At follow-up evaluation at an average of 5 years later, patients' mean scores had improved as follows: pain, from 2.3 to 5.7; function scores, from 2.3 to 4.5; mobility scores, from 2.3 to 4.4; Harris hip scores, from 37.8 to 95. We recommend both placing the cup in the true acetabulum to maximize host-bone contact with the implant and preserve as much host bone as possible and femoral-shortening osteotomy for a lower incidence of nerve injury than with aggressive soft-tissue release. PMID- 16376259 TI - Total hip arthroplasty in adult patients who had developmental dysplasia of the hip. AB - This study evaluated the hypothesis that the clinical results are equivalent in the group of patients with dysplasia, low dislocation, and high dislocation types using a contemporary technique for hip arthroplasty. The mean age of patients at the time of the index operation was 49.5 years (range, 29-61 years). Thirty patients (40 hips, 35%) had dysplasia, 22 (34 hips, 29%) had a low dislocation, and 24 (42 hips, 36%) had a high dislocation. The mean follow-up was 9.7 years (range, 6-14 years). In the high dislocation group, 7 hips (17%) had a revision of one or both component. In low dislocation group, 3 hips (9%) had a revision of one or both components. In the dysplastic group, 2 hips (5%) had a revision of one or both components. PMID- 16376260 TI - Health outcome measures in the evaluation of total hip arthroplasties--a comparison between the Harris hip score and the Oxford hip score. AB - There has been an increasing need for the sensitive and reproducible measurement of the outcome after hip surgery. Numerous hip scoring systems, varying in their complexity and disease specificity, have been designed to achieve a measure of outcome-some rely ultimately on the judgement of the surgeon, whereas others rely on the patients' perceptions. The Oxford hip score (OHS) has been found to be easier to administer and achieves a much higher follow-up rate than that of the Harris hip score (HHS). Unfortunately, with the availability of numerous scoring systems and the publication of data in these systems, it has been difficult to compare results. Our aim was to compare the more widely used HHS to the shorter OHS. We followed 200 consecutive primary total hip arthroplasties (196 patients between January 1994 and May 1995) for an average of 5 years. All patients had a preoperative HHS recorded. At the 5-year review, assessment was made using OHS and the HHS. There were 115 hips that had full OHS and HHS available. The mean OHS was 19.1 (range 12-52, SD 9.5), and HHS was 89.4 (range 47-100, SD 13.3) at follow-up. The Spearman correlation showed good negative correlation between the 2 results (Spearman rank -0.712, P < .0001). The HHS vs OHS shows good correlation at 5 years. This is the first study to confirm that correlation persists for the OHS in the medium term. We include a classification of OHS of excellent (<19), good (19-26), fair (27-33), and poor (>33) outcomes which correlate well with the HHS. This study enables the case for the Oxford data with its easier analysis and higher compliance rate to be used more directly to compare studies that use the HHS. PMID- 16376261 TI - Mechanisms for pumping fluid through cementless acetabular components with holes. AB - The pumping of fluid and polyethylene wear debris from the joint space to the retroacetabular bone is implicated in the pathogenesis of osteolysis. Three possible mechanisms for this pumping: pressure gradients, diaphragm pumping, and piston pumping were studied in vitro in a laboratory model. The simulated activities of rising from a chair and climbing stairs produced high-pressure gradients and high angles of loading that could pump fluid through the apical hole to the retroacetabular bone. A noncongruent liner acted as a diaphragm pump, producing pressures 6 times higher than that seen with a congruent liner. Pistoning motion of the liner produced pressures 8 times higher than when no pistoning occurs. These pumping mechanisms could be mitigated by the use of acetabular components without holes. PMID- 16376262 TI - Serum interleukin 8 levels correlate with synovial fluid levels in patients with aseptic loosening of hip prosthesis. AB - Levels of inflammatory cytokines (tumor necrosis factor alpha, interleukin [IL] 6, and IL-8) in serum from patients with osteolysis on radiographs after hip arthroplasty [osteolysis(+), n = 28], patients without osteolysis after hip arthroplasty [osteolysis(-), n = 24], and nonoperated healthy subjects [controls, n = 20] were determined. In addition, cytokine levels in synovial fluid from patients undergoing revision total hip arthroplasty (n = 14) for loosening were measured and compared with each other and with the area of osteolysis on radiographs. Serum IL-6 and IL-8 levels were significantly higher in the osteolysis(+) group than in the osteolysis(-) or the control groups. Furthermore, a significant correlation was found between the serum and synovial fluid IL-8 levels and between synovial fluid IL-8 levels and the area of osteolysis in patients undergoing revision total hip arthroplasty. Therefore, serum IL-8 levels could be a useful periprosthetic osteolysis marker. PMID- 16376263 TI - In vitro elution of vancomycin from calcium phosphate cement. AB - Antibiotic-impregnated bone cement beads have become popular for the treatment of osteomyelitis and/or prosthesis infection. However, bone cement has the disadvantage of heating up during polymerization of cement. Recently, calcium phosphate cement (CPC) has been used as a bone replacement and augmentation, and it does not heat up during polymerization. First, we measured the release rate of vancomycin (VCM) from bone cement and CPC impregnated with VCM for 2 weeks in vitro. The mean concentration of VCM for CPC was 62.6 times at 7 days (258 +/- 29 vs 4.12 +/- 1.0) and 6.7 times at 13 days (15.5 +/- 5.5 vs 2.3 +/- 0.7). Second, we were successful in treating 2 cases of osteomyelitis and prosthesis infection with VCM-impregnated CPC. From this study, we concluded that VCM-impregnated CPC might be an effective material for the treatment of osteomyelitis and/or prosthesis infection. PMID- 16376264 TI - The functional flexion-extension axis of the knee corresponds to the surgical epicondylar axis: in vivo analysis using a biplanar image-matching technique. AB - We investigated the concept that the knee has a fixed flexion-extension axis in the posterior femoral condyles and that this functional axis corresponds to the surgical epicondylar axis in vivo. We used a biplanar image-matching technique to perform the in vivo analysis of 9 normal knees to determine the location of the functional flexion-extension axis of the knee using an optimization technique. The functional flexion-extension axis passed through the sulcus of the medial epicondyle and the prominence of the lateral epicondyle. Flexion and extension of the knee could be represented as a rotation around a fixed axis, and this functional axis corresponded to the surgical epicondylar axis during a 0 degrees to 90 degrees flexion. This study assists more understanding of knee kinematics and provides useful information for the design and positioning of the prostheses used in total knee arthroplasty. PMID- 16376265 TI - Peroneal nerve palsy after total knee arthroplasty. PMID- 16376266 TI - Total hip arthroplasty in osteopetrosis using computer-assisted fluoroscopic navigation. AB - Hybrid total hip arthroplasty with computer-assisted fluoroscopic navigation was performed on a patient with osteoarthritis due to an autosomal dominant form of osteopetrosis. The surgical procedures were difficult because the bone was extremely hard and brittle with obliteration of the medullary cavity. Especially, preparation of a femoral canal for the stem was technically challenging and required changes from a conventional surgical procedure. Therefore, we used a computer-assisted fluoroscopic navigation system to create the femoral cavity for the stem, and an accurate placement of the prosthesis was achieved. Navigation guidance can be a useful tool when performing arthroplasty in patients with hip osteoarthritis associated with osteopetrosis. PMID- 16376267 TI - Flexion contracture after total knee arthroplasty in a patient with Parkinson's disease: successful treatment with botulinum toxin type A. AB - The few reports in the orthopedic literature that discuss outcomes after total knee arthroplasty in patients with Parkinson's disease cite mixed results. These patients are at increased risk for the development of flexion contracture, which has been shown to significantly worsen functional scores. The present report describes the development of a flexion contracture in a patient with Parkinson's disease after total knee arthroplasty. This contracture was successfully treated with manipulation under anesthesia and injections of botulinum toxin type A into the hamstring and gastrocnemius muscles, in conjunction with a static progressive extension orthosis and rigorous physical therapy. PMID- 16376268 TI - Bilateral gluteal compartment syndrome after total hip arthroplasty under epidural anesthesia. AB - The gluteal compartment syndrome is uncommon and is discussed in only a few published case reports. The simultaneous bilateral gluteal compartment syndrome is exceptionally rare and is tackled in only 4 case reports to date. We report a case of bilateral gluteal compartment syndrome after total hip arthroplasty under epidural anesthesia and discuss its management. PMID- 16376269 TI - Late fatigue fracture of a modern cemented [corrected] cobalt chrome stem for total hip arthroplasty: a report of 10 cases. AB - We report 10 fatigue fractures of a modern, cemented, cobalt chromium alloy stem (Osteonics Omnifit) for total hip arthroplasty occurring between 1995 and 2004. The primary total hip arthroplasties had been performed between 1989 and 1996. The average age at the time of surgery was 54 years (range, 34-70 years), and the average body mass index was 29 (range, 20-38). The time in situ of the prosthesis at the time of fracture averaged 8 years (range, 4-12 years). Intermediate follow up radiographs before the fracture were available in 7 cases, all of which demonstrated loss of calcar support. Scanning electron microscopy of the fracture surfaces in 3 of the components showed porosity near the initiation site. Metallography of polished and etched cross sections near the fracture surface revealed large grain size. In the presence of a spontaneous onset of thigh pain at intermediate follow-up in patients with this stem, particularly if associated with loss of calcar support, a fatigue fracture should be considered. PMID- 16376270 TI - Pharmacotherapy of spasticity in children with cerebral palsy. AB - Spasticity is one of the most common symptoms presented by neurologic patients. Apart from surgical management, drug therapy is an important treatment of children suffering from spasticity. In this review, recent advances in the pharmacologic armamentarium are reported in detail. In particular, there are oral medications (benzodiazepines, baclofen, dantrolene sodium, alpha 2 adrenergic agonists) and parenteral medications (botulinum toxin type A and B, alcohol). Moreover, there is also baclofen that can be administered intrathecally. There are some reports supporting the use of intramuscular alcohol (45% and/or 5-7% phenol) to reduce spasticity without the loss of voluntary movement or loss of sensation. Among these drugs, intrathecal baclofen is one of the most effective substances that can reduce spasticity significantly in the upper and lower extremities. Finally, the effectiveness of therapy with botulinum toxin type A in the management of spasticity is analyzed. Botulinum toxin type A reduces hypertonia in the injected muscles for a period of 2 to 4 months without important side effects. The purpose of this article is to provide an overview of available oral and parenteral drugs for treatment of spasticity in cerebral palsy and to outline indications and contraindications. PMID- 16376271 TI - Characterization of symptoms of sleep disorders in children with headache. AB - To investigate the prevalence of sleep disorders and their symptoms in children with headaches, 64 patients in the outpatient clinics of the University of Chicago Department of Pediatric Neurology were interviewed. Investigated disorders included excessive daytime sleepiness, narcolepsy, insomnia, sleep apnea, restlessness, and parasomnias. Unlike previous studies, subjects were compared with matched control patients by age and sex. Both headache and nonheadache groups completed a 111-item questionnaire detailing sleep symptoms and behaviors. It was found that children with headaches have a significantly higher prevalence of excessive daytime sleepiness, narcolepsy, and insomnia than children without headaches (P < 0.005), which is consistent with prior literature. A similar result was obtained in examining only migraines. However, we did not find a significantly higher prevalence of symptoms of sleep apnea, restlessness, and parasomnias, which contradicts previous literature. Also, the effect of medications taken by headache patients as a confounding factor was insignificant. Overall, pediatricians may find it beneficial to ask about daytime sleepiness, narcolepsy, and insomnia when treating a headache patient. PMID- 16376272 TI - Everyday cognitive function after craniopharyngioma in childhood. AB - Despite clinical impressions that cognitive complaints are prominent in patients with a history of craniopharyngioma, formal neuropsychologic documentation is inconsistent. This study assessed everyday cognitive complaints and neuropsychologic test performance to evaluate the prevalence of problems and the relationship of these domains to one another in patients treated for craniopharyngioma in childhood or adolescence. Ten patients treated for craniopharyngioma completed measures of everyday cognitive function (Cognitive Failures Questionnaire, Rivermead Behavioural Memory Test) and a battery of standard neuropsychologic tests. The prevalence of problems was ascertained for each measure. Most patients demonstrated significant deficits in everyday memory (Cognitive Failures Questionnaire, 9/10 patients; Rivermead Behavioural Memory Test, 7/10 patients). Scores were within normal limits, however, for intelligence quotient, achievement, attention, verbal memory, and spatial working memory. Processing speed was slow (5/10 patients). Spatial working memory predicted Cognitive Failures Questionnaire (P < 0.07), as did somatic symptoms from the Beck Depression Inventory (P < 0.01), but these associations appeared independent. Adolescents and young adults with treated craniopharyngioma experience deficits in everyday cognitive functions, many involving memory, that are not easily detected by standard neuropsychologic testing. The extent of self rated cognitive problems is related to spatial working memory and somatic concerns. PMID- 16376273 TI - New proteins configure a brain drug resistance map in tuberous sclerosis. AB - Epileptogenic cortical tubers, characterized by dysplastic neurons and balloon cells, is a frequent feature of tuberous sclerosis. In severe tuberous sclerosis affected individuals, seizures are refractory to medication. Multidrug resistance proteins (multidrug resistance protein-1 [MDR-1] and multidrug resistance associated protein-1 [MRP-1]) have been found to be highly expressed in epileptogenic cortical tubers. However, two new proteins related to refractoriness in cancer (breast cancer resistance protein and major vault protein) have not been investigated in tuberous sclerosis and refractory epilepsy. On the same brain specimens previously describing the MDR-1 and MRP-1 expression, we investigated retrospectively breast cancer resistance protein and major vault protein by specific monoclonal antibodies and routine immunohistochemistry methods. Breast cancer resistance protein was present in vascular endothelial cells from all the vessels examined in 3 of 3 cases. Major vault protein was detected in only one case, and selectively expressed in several but not all ballooned cells. In epileptogenic cortical tubers, the additional expression of breast cancer resistance protein on vessels and major vault protein in some ballooned cells to the previously demonstrated expression of MDR-1 and MRP-1 (in vessels, astroglia, microglia, neurons, and ballooned cells) configures a brain protein pharmacoresistance map from patients with tuberous sclerosis and refractory epilepsy. PMID- 16376274 TI - TNF-alpha and IL-6 in the diagnosis of bacterial and aseptic meningitis in children. AB - The objective of this study was to analyze the usefulness of tumor necrosis factor-alpha and interleukin-6 cerebrospinal fluid concentrations for the differential diagnosis between bacterial and aseptic meningitis in children and in the prognostic evaluation. A cross-sectional study was performed on 35 children between 1 month and 12 years of age with suspected meningitis. Cytokines determination was performed by enzyme-linked immunosorbent assay technique. The Mann-Whitney test and Spearman's correlation coefficients were used for statistical analysis. Six children presented bacterial meningitis, 13 aseptic, and 16 had no meningitis. The tumor necrosis factor-alpha concentrations were significantly higher in the bacterial meningitis group as compared with the aseptic group (P = 0.001) and among groups with and without meningitis (P = 0.000). There was correlation between tumor necrosis factor-alpha and cerebrospinal fluid leukocytes (P = 0.019), protein (P = 0.000), and glucose (P = 0.038). There was no association between cytokines and complications of the meningitis. The tumor necrosis factor-alpha concentrations in the cerebrospinal fluid were useful markers for distinguishing bacterial from aseptic meningitis and were demonstrated to be useful in evaluating the intensity of the inflammatory process in the central nervous system. PMID- 16376275 TI - Amplitude spectral analysis of theta/alpha/beta waves in preterm infants. AB - The aim of this study is to clarify the usefulness of amplitude spectral analysis for the evaluation of maturational changes of theta, alpha, and beta activities in preterm infants. We chose 10 healthy infants, who were born without complications other than prematurity, who were 29-30 weeks, 31-32 weeks, and 33 34 weeks of postconceptional age, respectively, at time of electroencephalogram recordings. Fast Fourier transform algorithm was applied for amplitude spectral analysis. The analyzed data were divided into five frequency bands: theta 1 4-6 Hz, theta 2 6-8 Hz, alpha 8-13 Hz, beta 1 13-20 Hz, and beta 2 20-30 Hz. The average amplitude of six segments with continuous patterns with high-voltage slow waves of preterm infants was calculated in each frequency band. A significant reduction of the amplitude along with postconceptional age was present in all leads in theta 1 and theta 2 bands. On the other hand, a significant correlation with postconceptional age was not observed in alpha, beta 1, or beta 2 bands. In conclusion, maturational electroencephalographic changes assessed by amplitude spectral analysis were prominent in theta activities and were not remarkable in alpha or beta activities. PMID- 16376276 TI - Magnetic resonance imaging and proton magnetic resonance spectroscopy of megalencephaly and dilated Virchow-Robin spaces. AB - Megalencephaly with dilated Virchow-Robin spaces has been suggested to represent a new clinical entity. This report describes two males and a female who have been monitored from pregnancy. The patients manifest a relatively normal psychomotor development with some minor neurologic symptoms such as mild muscle hypotonia and clumsy motor performance. Biochemical and electrophysiologic tests were normal. In the white matter of the brain, a prominent dilatation of the Virchow-Robin spaces with some adjacent signal alterations could be demonstrated by magnetic resonance imaging. Magnetic resonance spectroscopy revealed normal metabolite concentrations in the cortical and deep gray matter and normal-appearing white matter. Affected white matter was characterized by mildly reduced to normal levels of myo-inositol and a decrease of all other metabolites including total N acetyl moieties, choline-containing compounds, and total creatine. These data indicate that the dilatation of Virchow-Robin spaces reflects an underlying brain pathology causing neuroaxonal damage. Possible differential diagnoses are discussed. PMID- 16376277 TI - Congenital muscular torticollis: evaluation and classification. AB - In this investigation of congenital muscular torticollis, 311 infants treated consecutively for congenital torticollis over an 8-year period (1995-2003) at the Pediatric Neurology Clinic of Istanbul Medical Faculty, Istanbul University, Turkey were reviewed retrospectively. The clinical presentation, associated abnormalities, treatment, and outcomes of the overall group and of subgroups divided according to an ultrasonography-based classification were evaluated. All patients were evaluated using a standard approach: cervical ultrasonography was performed, and the patients were divided into two subgroups. Each group was scanned for other anomalies, and outcomes were compared. The mean age at diagnosis was 2.3 months; patients included in this study were 138 males and 173 females. Two clinical subgroups, comprised of sternomastoid tumors 85% and postural torticollis 15%, were identified. Passive range of motion was the initial treatment recommended for all of the patients. Follow-up data were available for all 311 patients; 95% experienced total resolution and 5% experienced subtotal resolution. We conclude that the majority of children with congenital muscular torticollis experience total resolution of symptoms. The success rate of conservative treatment is primarily dependent on the patients' age at the initiation of exercises and ultrasonographic findings. PMID- 16376278 TI - Vestibular neuritis caused by enteroviral infection. AB - Vestibular neuritis is characterized by the sudden onset of nausea, vomiting, and spontaneous horizontal or horizonto-rotatory nystagmus. The etiology of the disease is multifactorial. Mumps, rubella, herpes simplex virus type 1, cytomegalovirus, and Epstein-Barr virus may have a role in the disease. Enteroviruses are among the other rare causes. This report presents a 7-year-old male admitted with nausea, vomiting, rotatory vertigo, horizonto-rotatory nystagmus with positive Romberg's sign and positive head-thrust test. Cranial magnetic resonance imaging and audiometry of the patient were normal. He was diagnosed with vestibular neuritis, and steroid therapy was initiated. At the second month of follow-up, all symptoms had regressed. To the best of our knowledge, this case report describes the first pediatric patient in whom enteroviral ribonucleic acid is documented both in cerebrospinal fluid and in nasopharyngeal material in active disease. This finding supports the possible role of enteroviruses in the etiology of vestibular neuritis. PMID- 16376279 TI - Focal glucose hypermetabolism in interictal state of West syndrome. AB - This report concerns two siblings from a tetrad, both of whom had West syndrome with atypical findings on positron emission tomography using [(18)F] fluorodeoxyglucose. One manifested periventricular leukoencephalopathy, and the other had periventricular leukoencephalopathy as well as porencephaly because of fetal distress and brain parenchymal hemorrhage in the neonatal period. They developed West syndrome at the age of 9 months. Fluorodeoxyglucose-positron emission tomography study performed after cessation of their seizures revealed an increase in glucose metabolism. The corresponding region presented low-level accumulation in [(11)C]flumazenil positron emission tomography. The patients remained seizure-free for more than 1 month, and their electroencephalograms only occasionally disclosed sporadic paroxysmal discharges. Because of the decreased density of benzodiazepine receptor in these lesions, the activity of the excitatory neuron system may overexpress that of the inhibitory neuron system, thus resulting in epileptogenesis of the lesions. It is suggested that fluorodeoxyglucose and flumazenil-positron emission tomography revealed functional abnormalities and that epileptogenesis of these patients is still active even when the patient is seizure-free and there are mild epileptogenic discharges on electroencephalogram. PMID- 16376280 TI - Hypomelanosis of Ito associated with precocious puberty. AB - Hypomelanosis of Ito has been associated with precocious puberty in two cases. This study reports a third case involving a female with hypomelanosis of Ito including severe mental retardation and seizure disorder with autonomic symptoms (gastroesophageal reflux and asthma). At age 5 she developed vaginal discharge, thelarche, and adrenarche. Soon after, she died of pneumonia stemming from her neurologic deficits. A postmortem examination revealed normal gonads and endocrine organs. The breasts and vulva were prematurely developed. Central nervous system findings included megalencephaly, neuronal eosinophilic inclusions, leptomeningeal neuroglial heterotopias, and cortical dysplasia. Both previously reported cases of hypomelanosis of Ito associated with precocious puberty had abnormal gonads and responded to cyproterone acetate therapy, indicating a peripheral mechanism of precocious puberty (gonadotrophin independent). The current case, which was autopsied, lacks significant gonadal pathology, and has extensive neurologic involvement that suggests that a central mechanism of precocious puberty (gonadotrophin-dependent) can also be associated with hypomelanosis of Ito. PMID- 16376281 TI - Levetiracetam in a neonate with malignant migrating partial seizures. AB - This report describes the first neonatal case of "malignant migrating partial seizures in infancy" with a positive therapeutic response to levetiracetam. This patient is the youngest reported infant with this rare syndrome, and the report provides the first documentation on levetiracetam treatment in a neonatal patient. Treatment with levetiracetam improved both ictal and interictal status. This observation also highlights the need to consider and include malignant migrating partial seizures in the differential diagnosis of early neonatal seizure disorders, even during the first hours of life. PMID- 16376282 TI - Immunoglobulin for concurrent Guillain-Barre and immune thrombocytopenic purpura. AB - Guillain-Barre syndrome, or acute inflammatory demyelinating polyradiculoneuropathy, and immune thrombocytopenic purpura are both autoimmune disorders thought to result from molecular mimicry in response to an antecedent introduction of foreign antigen. Guillain-Barre syndrome is an ascending motor paralysis that can lead to respiratory compromise. Immune thrombocytopenic purpura is an isolated disorder of platelet destruction leading to mucocutaneous bleeding. Guillain-Barre does not typically occur with other autoimmune disorders, and concurrent Guillain-Barre and immune thrombocytopenic purpura has only rarely been reported. We present a patient with both conditions who experienced prompt resolution of neurologic and hematologic sequelae after intravenous immunoglobulin therapy was initiated within 12 hours of presentation. The case provides further evidence that Guillain-Barre syndrome and immune thrombocytopenic purpura can occur simultaneously, possibly caused by a similar pathogenic mechanism, as well as suggesting that the prompt initiation of intravenous immunoglobulin is an effective monotherapy leading to prompt resolution of both conditions and prevention of further sequelae. PMID- 16376283 TI - Very-high-dose phenobarbital for childhood refractory status epilepticus. AB - This report describes the management of three children, ranging in age from 7 to 9 years, who had refractory status epilepticus. This condition was likely the result of viral encephalitis; it manifested initially as generalized tonic-clonic seizure and later frequent subtle eye staring and twitching of mouth or limbs as well as tachycardia and dilatation of pupils. The seizures were not responsive to treatment with conventional doses of intravenous phenobarbital, phenytoin, and midazolam infusion. Very-high-dose phenobarbital was administered at accumulated daily doses up to 80 mg/kg, with a resulting serum level of more than 1000 mumol/L. It was effective in achieving seizure control, with milder adverse effects compared with thiopental infusion. PMID- 16376284 TI - Septo-optic dysplasia complex: a heterogeneous malformation syndrome. AB - Septo-optic dysplasia is defined by a variable combination of dysgenesis of midline brain structures including optic nerve hypoplasia and hypothalamic pituitary dysfunction often associated with a wide variety of brain malformations of cortical development. Multiple congenital anomalies have been reported only sporadically. Despite recent demonstration of the possible pathogenic role of HESX1/Hesx1 gene (a homeobox gene important for development of prosencephalon), the etiology of most cases of septo-optic dysplasia still remains unclear. This report describes eight children (4 males, 4 females; age 2 to 17 years) with septo-optic dysplasia who manifested dysmorphic features (involving not only the midline facial structures) and a spectrum of additional clinical and imaging features including autism, facial hemangioma, and holoprosencephaly. Full mutational screening for the HESX1 gene in seven of eight children was negative. Based on the extreme variability of the clinical and imaging phenotypes hereby observed, on literature review, and on septo-optic dysplasia animal models, this study confirmed that the phenotypic heterogeneity in septo-optic dysplasia is high. We suggest that: (1) dysmorphic features are more frequent than previously thought--they may represent a relevant part of the phenotype; (2) septo-optic dysplasia should be recategorized as an heterogeneous malformation syndrome (septo-optic dysplasia complex) (encompassing multiple brain, endocrine, and systemic anomalies) rather than a single precisely defined entity. PMID- 16376285 TI - Hypertension-induced cerebellar encephalopathy and hydrocephalus in a male. AB - Hypertensive encephalopathy is believed to be caused by an abrupt elevation in systemic blood pressure. It rarely occurs in children and can be neurologically devastating if it is not recognized and treated immediately. This report describes an 11-year-old male who presented with edema and a cerebellar lesion, with acute obstructive hydrocephalus resulting from hypertensive encephalopathy. A shunt was inserted to relieve pressure in the acute stage. The patient's hydrocephalus and cerebellar swelling subsided when his blood pressure was controlled. The cerebellar lesion had been initially diagnosed as a glioma. In children, a cerebellar lesion occurring with acute obstructive hydrocephalus and hypertensive encephalopathy is rare but reversible. Clinicians should be aware of this condition because it might be misdiagnosed as a tumor of the posterior fossa. PMID- 16376286 TI - Monozygotic twins discordant for Klippel-Feil syndrome. AB - In a pair of Japanese monozygotic twins, one manifested Klippel-Feil syndrome, a short neck with C(1-4) vertebra fusion, whereas the other was normal. The discordance between the twins suggests that Klippel-Feil syndrome results in part from a postzygotic somatic mutation or intrauterine environmental factors. PMID- 16376287 TI - Immunoaffinity extraction of a peptide modified by a small molecule. AB - We investigated the affinity extraction conditions required to isolate peptide fragments modified with small molecules using an antibody that has a high affinity for the target small molecule. Investigation of antibody conformation and the retention behavior of the modified peptides on an immunosorbent matrix demonstrated the importance in efficient extraction of both the dissociation of hydrophobic interactions and the breakdown of the antibody conformation. Hydrophobic interactions, which anchor the small ligand to the paratope, were retained even when the three-dimensional structure of the antibody disintegrated in an acidic solution. For efficient extraction of a target peptide modified by a small molecule, it is therefore important to use an acidic solvent containing an organic modifier such as methanol at a concentration greater than 40% (v/v). We demonstrated the feasibility of this immunoaffinity extraction by application of this procedure to the analysis of modified peptide fragments obtained from a digestion of human serum albumin. The peptide fragments were affinity labeled with chenodeoxycholyl adenylate for analysis of the chenodeoxycholate binding site. This purification method could isolate the low levels of modified peptide contained in the reaction mixture, despite the presence of appreciable quantities of unlabeled peptide fragments. PMID- 16376288 TI - Inhibition assay of beta-hematin formation initiated by lecithin for screening new antimalarial drugs. AB - Measurement of heme crystallization provides a tool for screening new antimalarial drugs. Current assays for heme crystallization have employed initiators such as thermo, histidine-rich proteins, and lipids extracted from parasites and infected plasma. These initiators are unnatural or require laborious steps to prepare. In this study, we used a commercially available lipid, lecithin, a kind of phospholipid containing about 50% unsaturated fatty acids, as an initiator for heme crystal (beta-hematin) formation. We demonstrated that the inhibition of lecithin-based beta-hematin formation by antimalarial drugs is highly correlated with the preformed beta-hematin-based method. In addition, the lecithin-based assay is sensitive and convenient for large-scale screening of new novel antimalarials. We also indicated that dimethyl sulfoxide is an ideal solvent for preparation of heme stock solution, which is stable and can be used for 1 month. PMID- 16376289 TI - Single-step purification of myristoylated and nonmyristoylated recoverin and substrate dependence of myristoylation level. AB - Recoverin is cotranslationally modified by the covalent linkage of a myristoyl group to its N terminus. It is a member of a family of Ca(2+)-myristoyl switch proteins. Recombinant myristoylated revoverin is currently produced by the cotransformation of bacteria with recoverin and an enzyme that allows N myristoylation and by supplementing the culture medium with myristic acid. A large variation in the myristoylation level of recoverin and in the amount of myristic acid supplied to the culture medium can be found in the literature. Moreover, although it is known to strongly affect bacterial growth, the amount of ethanol used to solubilize myristic acid is only scarcely mentioned. To improve our understanding of the parameters responsible for recombinant recoverin myristoylation, the effects of myristic acid and ethanol on recoverin myristoylation and expression levels have been systematically studied. In addition, a single-step purification procedure to produce purified myristoylated and nonmyristoylated recombinant recoverin has also been devised. Finally, sodium myristate has been used as an efficient alternative substrate to achieve high myristoylation and expression levels of recoverin. Given that a large number of proteins are myristoylated, these procedures could be applied to several other proteins in addition to recoverin. PMID- 16376290 TI - Best literature values for the pK of carbonic and phosphoric acid under physiological conditions. PMID- 16376291 TI - Biting efficiency in relation to incisal angulation. AB - Previous studies suggest the orientation of the incisor teeth to the path of jaw movement pronouncedly affects their efficiency of action. To test this hypothesis, upper and lower incisal portions of a study model with ideal Asian occlusion were mounted on a mechanical tester and impressed into rectangular blocks made of 15.3% w/w starch gels or Cheddar cheese. At varying angulations of attack between the long axis of the teeth (defining the orientation of the tooth crown) and force direction, the teeth were driven into the blocks for 4-6mm until fractures had grown in the foods. Both the angle of attack and the work divided by fractured area produced in the food, termed 'work to fracture', were measured. The food type (cheese or gel), incisal type (upper or lower pair) and angle of inclination were significant effects on the works to fracture. The minimal work was for angles slightly proclined to the vertical, although only angles of proclination >40 degrees were significantly higher than all other angles. Retroclination or large angles of proclination made little difference to the work done, but produced markedly off-axis cracks. It is suggested that human incisors act most efficiently at small gapes and that orthodontic corrections will thus offer definite functional benefits. PMID- 16376292 TI - Osmotic pressure and vasculature of gingiva in periodontal disease: an experimental study in rats. AB - Alterations in vascularisation, vasodilatation and tissue osmotic pressure (OP) are inevitable aspects of the inflammatory process that have an adverse effect on the fluid dynamics of the tissue involved. The aim of this study was to investigate tissue OP and its relationship with the vasculature in inflammed gingival tissues, in order to reveal the possible effects of vascular changes on OP in the fluid dynamics of periodontal soft tissues during periodontal disease. The parameters of fluid dynamics assessed in this study were OP, vascularisation and vasodilatation. Ligature-induced periodontitis was performed in 10 rats (test group), and gingival biopsies taken from the diseased teeth were utilised for the test procedures. These biopsies were compared with biopsies of the same teeth from 10 periodontally healthy rats (control group). OP was measured in mosmol/kg using a semi-micro digital osmometer. Vascularisation and vasodilatation were examined histopathologically; the number of vessels (VN) was quantified and the micrometric changes in vessel diameters (VD) were calculated as the alterations in the vasculature. OP, VN and VD were found to be higher in the test group (84.3+/-37.1 mosmol/kg, 13.2+/-3.2 and 19.5+/-1.3 microm, respectively) than the control group (11.6+/-3.8 mosmol/kg, 6.8+/-1.1 and 15.5+/-2.4 microm, respectively) (P<0.000). There was a strong, positive correlation between OP and VN (r=0.55, P<0.000) and a weak, negative correlation between OP and VD (r=0.1, P>0.05) in the test group. These results confirm that the OP of periodontal soft tissues does change during inflammatory conditions. The increase in OP during this process may be affected by increased vascularisation in the inflammed tissue. PMID- 16376293 TI - Proton transfers in the bacteriorhodopsin photocycle. AB - The steps in the mechanism of proton transport in bacteriorhodopsin include examples for most kinds of proton transfer reactions that might occur in a transmembrane pump: proton transfer via a bridging water molecule, coupled protonation/deprotonation of two buried groups separated by a considerable distance, long-range proton migration over a hydrogen-bonded aqueous chain, and capture as well as release of protons at the membrane-water interface. The conceptual and technical advantages of this system have allowed close examination of many of these model reactions, some at an atomic level. PMID- 16376294 TI - Lysophospholipid and fatty acid inhibition of pulmonary surfactant: non-enzymatic models of phospholipase A2 surfactant hydrolysis. AB - Secretory A(2) phospholipases (sPLA(2)) hydrolyze surfactant phospholipids cause surfactant dysfunction and are elevated in lung inflammation. Phospholipase mediated surfactant hydrolysis may disrupt surfactant function by generation of lysophospholipids and free fatty acids and/or depletion of native phospholipids. In this study, we quantitatively assessed multiple mechanisms of sPLA(2)-mediated surfactant dysfunction using non-enzymatic models including supplementation of surfactants with exogenous lysophospholipids and free fatty acids. Our data demonstrated lysophospholipids at levels >or=10 mol% of total phospholipid (i.e., >or=10% hydrolysis) led to a significant increase in minimum surface tension and increased the time to achieve a normal minimum surface tension. Lysophospholipid inhibition of surfactant function was independent of the lysophospholipid head group or total phospholipid concentration. Free fatty acids (palmitic acid, oleic acid) alone had little effect on minimum surface tension, but did increase the maximum surface tension and the time to achieve normal minimum surface tension. The combined effect of equimolar free fatty acids and lysophospholipids was not different from the effect of lysophospholipids alone for any measurement of surfactant function. Surfactant proteins did not change the percent lysophospholipids required to increase minimum surface tension. As a mechanism that causes surfactant dysfunction, depletion of native phospholipids required much greater change (equivalent to >80% hydrolysis) than generation of lysophospholipids. In summary, generation of lysophospholipids is the principal mechanism of phospholipase-mediated surfactant injury in our non-enzymatic models. These models and findings will assist in understanding more complex in vitro and in vivo studies of phospholipase-mediated surfactant injury. PMID- 16376295 TI - Hiding anxiety versus acknowledgment of anxiety in social interaction: relationship with social anxiety. AB - According to cognitive and interpersonal models, safety behaviors in social phobia (e.g., avoiding eye contact, hiding blushing) erroneously induce negative evaluation by interaction partners. Presumably, a bias about the social outcome of safety behaviors causes this negative interaction cycle. Such a bias might be subject to double standard in social phobia (i.e., having more stringent rules for oneself than for others). Female students (n=81) predicted more negative social outcomes for a prominent type of safety behavior, hiding anxiety, than for the opposite behavior, acknowledgment of anxiety and ongoing behavior (control condition) in scripts of self- and other-target persons. The relation between social anxiety and double standard was robust. Social anxiety did not relate to a cognitive bias regarding hiding ones anxiety, as we expected, but was associated with the belief that acknowledgment of anxiety has negative social outcomes specifically for them. These results are evaluated in light of the interpersonal consequences anxiety-related behaviors have in social interactions. PMID- 16376296 TI - A cognitive behavioral workshop for subclinical obsessions and compulsions. AB - The present study investigates the effectiveness of a 3-h cognitive behavioral workshop for individuals, ages 18-22, with subclinical obsessions and compulsions. It was hypothesized that, compared to individuals in an assessment only waitlist group (n = 42), individuals assigned to the workshop group (n = 43) would experience a significant decrease in obsessive-compulsive (OC) symptomatology, comorbidity, and thought action fusion endorsement at 1-month and 5-month follow-up assessments. An additional outcome of interest was the number of incident cases of obsessive-compulsive disorder (OCD) over the course of the study. The results indicated that the workshop group reported a significantly fewer number of OC symptoms at 5-month follow-up and endorsed significantly less thought action fusion at both follow-up points. However, there were no differences between groups in severity of OC symptoms and number of comorbid diagnoses endorsed. Only one incident case of OCD was observed during the study, from a participant in the waitlist group. These results are discussed in reference to treatment of subclinical anxiety symptoms. PMID- 16376297 TI - A fibronectin-binding protein from Streptococcus equi binds collagen and modulates cell-mediated collagen gel contraction. AB - The N-terminal fragment (FNZN) of the fibronectin-binding protein FNZ from Streptococcus equi subspecies zooepidemicus was investigated as to effects on murine cell interactions with extracellular matrix proteins. FNZN bound to immobilized fibronectin (FN) and native, but not denatured, collagen type I. FNZN had no effect on primary adhesion of cells from the murine myoblastic C2C12 cell line to immobilized fibronectin. C2C12 cells adhered to immobilized FNZN, a process that was not inhibited by anti-human FN IgG or by an inhibitor of integrin alphaVbeta3. C2C12 cells lack collagen-binding beta1 integrins and neither adhere to native collagen nor mediate contraction of three-dimensional collagen gels. FNZN stimulated collagen gel contraction by C2C12 cells but not adhesion of C2C12 cells to collagen. Experiments with an alphaVbeta3-inhibitor suggested that FNZN promoted contraction by a process requiring alphaVbeta3. Our data suggest that FNZN by binding to cells, collagen, and FN modulate complex adhesive processes mediated by the alphaVbeta3 integrin. Since alphaVbeta3 mediated contractile events function to counteract edema formation during inflammation, it is possible that FNZN and its secreted homologue FNE modulate edema responses in infected tissues. PMID- 16376298 TI - Streptozotocin inhibits O-GlcNAcase via the production of a transition state analog. AB - Streptozotocin (STZ) is a 2-deoxy-d-glucopyranose derivative of a class of drugs known as alkylnitrosoureas, and is an established diabetogenic agent whose cytotoxic affects on pancreatic beta-cells has been partially explained by the presence of its N-methyl-N-nitrosourea side chain, which has the ability to release nitric oxide as well as donate methyl groups to nucleotides in DNA. It has also been observed that STZ administration results in a rise in the level of O-GlcNAcylated proteins within beta-cells. Not coincidentally, STZ has also been shown to directly inhibit the O-GlcNAcase activity of the enzyme NCOAT in vitro, which is the only enzyme that possesses the ability to remove O-GlcNAc modifications on proteins in the nucleus and cytosol. Since O-GlcNAc modification plays a role on a number of proteins in a vast amount of cellular processes, this shift in whole-cell protein O-GlcNAcylation state affords another source of cell death. We set about to find the exact mechanism by which STZ inhibits O-GlcNAcase activity. Inhibition is achievable because the GlcNAc analog STZ targets the active site of the enzyme whereby it is catalyzed. During this process, the enzyme converts STZ to a compound that closely resembles the natural ligand transition state, but is distinctly more stable energetically. As a result, this analog is catalyzed to completion at a much slower rate, thereby out-competing GlcNAc substrate for the active site, and inhibiting the enzyme. PMID- 16376299 TI - A binding site for Pur alpha and Pur beta is structurally unstable and is required for replication in vivo from the rat aldolase B origin. AB - The rat aldolase B promoter acts as a replication origin in vivo, as well as an autonomously replicating sequence (ARS). Here, we examined roles of a polypurine stretch (site PPu) in this origin, which is indispensable to the ARS activity. Purification of site PPu-binding protein revealed that site PPu binds Puralpha and Purbeta, i.e., single-stranded DNA-binding proteins whose roles in replication have been implicated, but less clear. Biochemical analyses showed that site PPu even in a longer DNA fragment is unstable in terms of double-helix, implying that Puralpha/beta may stabilize single-stranded state. Deletion of site PPu from the origin DNA, which was ectopically positioned in the mouse chromosome, significantly reduced replicator activity. Chromatin immunoprecipitation experiments showed that deletion of site PPu abolishes binding of the Puralpha/beta proteins to the origin. These observations suggest functional roles of site PPu and Puralpha/beta proteins in replication initiation. PMID- 16376300 TI - A voltage-activated proton current in human cardiac fibroblasts. AB - A voltage-activated proton current in human cardiac fibroblasts, measured using the whole-cell recording configuration of the patch-clamp technique, is reported. Increasing the pH of the bathing solution shifted the current activation threshold to more negative potentials and increased both the current amplitude and its rate of activation. Changing the pH gradient by one unit caused a 51mV shift in the reversal potential of the current, demonstrating a high selectivity for protons of the channel carrying the current. Extracellularly applied Zn(2+) reversibly inhibited the current. Activation of the current contributes to the resting membrane conductance under conditions of intracellular acidosis. It is proposed that this current in cardiac fibroblasts is involved in the regulation of the intracellular pH and the membrane potential under physiological conditions as well as in response to pathological conditions such as ischemia. PMID- 16376301 TI - Correlation of PIK3Ca mutations with gene expression and drug sensitivity in NCI 60 cell lines. AB - The gene that encodes the alpha-isoform of phosphatidylinositol 3-kinase (PIK3Ca) is frequently mutated in human cancers. We profiled the mutation status of the PIK3Ca gene in the National Cancer Institute (NCI)-60 panel of human cancer cell lines maintained by the Developmental Therapeutics Program of the NCI. Mutation hotspots on the gene were PCR amplified and sequenced, and the trace data were analyzed with software designed to detect mutations. Seven of the cell lines tested have PIK3Ca mutations: two lines derived from breast cancer, two from colon cancer, two from ovarian cancer, and one from lung cancer. BRAF and EGFR genes were normal in the PIK3Ca mutant lines. Two of the cell lines with mutant PIK3Ca also have a mutant version of the KRAS gene. The mutation status was correlated with array-based gene expression that is publicly available for the NCI-60 cell lines. We found increased expression levels for estrogen receptor (ER) and ERBB2 in PIK3Ca mutant lines. The PIK3Ca mutation status was also correlated with compound screening data for the cell lines. PIK3Ca-mutant cell lines were relatively more sensitive than PIK3Ca-normal cell lines to the ER inhibitor tamoxifen and the AKT inhibitor triciribine, among other compounds. The results provide insights into the role of mutant PIK3Ca in oncogenic signaling and allow preliminary identification of novel targets for therapeutic intervention in cancers harboring PIK3Ca mutations. PMID- 16376302 TI - Altered NMDA receptor trafficking contributes to sleep deprivation-induced hippocampal synaptic and cognitive impairments. AB - Recent evidence indicates that continuous wakefulness (sleep deprivation, SD) causes impairments in behavioral performance and hippocampal long-term potentiation (LTP) in animals. However, the mechanisms by which SD impairs long term synaptic plasticity and cognitive function are not clear. Here, we report that 24-h SD in mice results in impaired hippocampus-dependent contextual memory and LTP and, unexpectedly, in reductions of the surface expression of NMDA receptor (NMDAR) subunit NR1 and NMDAR-mediated excitatory post-synaptic currents at hippocampal perforant path-dentate granule cell synapses. The results suggest that the reduction of functional NMDAR in hippocampal neurons may underlie the SD induced deficits in hippocampus-dependent contextual memory and long-term synaptic plasticity. PMID- 16376303 TI - Characterization of ARD1 variants in mammalian cells. AB - Mouse ARD1 (mARD1) has been reported to negatively regulate the hypoxia-inducible factor 1alpha (HIF-1alpha) protein by acetylating a lysine residue and enhancing HIF-1alpha ubiquitination and degradation. However, it was recently reported that human ARD1 (hARD1) does not affect HIF-1alpha stability. To further explore the activities of the two orthologs, three mouse (mARD1(198), mARD1(225), mARD1(235)) and two human (hARD1(131), hARD1(235)) variants were identified and characterized. Among these, mARD1(225) was previously reported as a novel negative regulator of HIF-1alpha. Amino acid sequence analysis showed that the C terminal region (aa 158-225) of mARD1(225) completely differs from those of mouse and human ARD1(235), although all three proteins share a well-conserved N acetyltransferase domain (aa 45-130). The effects of ARD1 variants were evaluated with respect to HIF-1alpha stability and acetylation activity. Interestingly, mARD1(225) strongly decreased the level of HIF-1alpha and increased the extent of acetylation, whereas mARD1(235) and hARD1(235) variants had a much weaker effect on HIF-1alpha stability and acetylation. These results suggest that ARD1 variants might have different effects on HIF-1alpha stability and acetylation, which may reflect diverse biological functions that remain to be determined. PMID- 16376304 TI - Cloning and genomic characterization of sytdep, a new synaptotagmin XIV-related gene. AB - We have identified a new human gene coined sytdep (synaptotagmin XIV-derived protein) in human neutrophils. Sytdep encodes a 188-amino acid sequence with a 21.435kDa deduced molecular mass, showing 75% identity to human synaptotagmin (syt) XIV. Human neutrophils express sytdep, but not syt XIV. Sytdep was upregulated during HL-60 neutrophil differentiation. Sytdep gene is located in human chromosome 4 and contains a unique exon, whereas syt XIV gene, located in chromosome 1, comprises 10 exons with 9 introns. Mouse genome did not contain sytdep. The N-terminal region of sytdep shows no homology with any known protein and, unlike synaptotagmin XIV isoforms, sytdep shows a unique C-terminal C2B domain. Polyclonal antibodies against the C2B domain of syt XIV recognized sytdep as a 27-kDa protein in human neutrophils. Genomic analyses suggest that human sytdep could derive from a retrotranslocation of a syt XIV transcript into chromosome 4. PMID- 16376305 TI - Perinuclear localisation of cellular retinoic acid binding protein I mRNA. AB - Retinoids are important metabolic and developmental regulators that act through nuclear receptors. The cellular retinoic acid binding protein CRABPI has been suggested to play a role in trafficking of retinoic acid but its exact functions and subcellular localisation remain unclear. Here we show that in CHO cells both exogenous CRABPI transcripts and tagged CRABPI protein have a perinuclear distribution that depends upon the 3'UTR of the mRNA. The CRABPI 3'UTR conferred perinuclear localisation on globin reporter transcripts. Deletion analysis indicated that the first 123nt of CRABPI 3'UTR are necessary for localisation of both CRABPI mRNA and protein. We propose that CRABPI mRNA is localised by a signal within its 3'UTR and that this partly determines the distribution of CRABPI protein. PMID- 16376306 TI - Inhibition of RNAse A family enzymes prevents degradation and loss of silencing activity of siRNAs in serum. AB - Small interfering RNAs (siRNA), RNA duplexes of approximately 21 nucleotides, offer a promising approach to specifically degrade RNAs in target cells by a process termed RNA interference. Insufficient in vivo-stability is a major problem of a systemic application of siRNAs in humans. The present study demonstrated that RNAse A-like RNAses degraded siRNAs in serum. The susceptibility of siRNAs towards degradation in serum was strongly enhanced by local clustering of A/Us within the siRNA sequence, i.e. regions showing low thermal stability, most notably at the ends of the molecule, and by 3' overhanging bases. Importantly, inhibition of RNAse A family enzymes prevented the degradation and loss of silencing activity of siRNAs in serum. Furthermore, the degradation of siRNAs was considerably faster in human than in mouse serum, suggesting that the degradation of siRNAs by RNAse A family enzymes might be a more challenging problem in a future therapeutic application of siRNAs in humans than in mouse models. Together, the present study indicates that siRNAs are degraded by RNAse A family enzymes in serum and that the kinetics of their degradation in serum depends on their sequence. These findings might be of great importance for a possible future human therapeutic application of siRNAs. PMID- 16376307 TI - Overcoming methotrexate resistance in breast cancer tumour cells by the use of a new cell-penetrating peptide. AB - Resistance to chemotherapy limits the effectiveness of anti-cancer drug treatment. Here, we present a new approach to overcome the setback of drug resistance by designing a conjugate of a cell-penetrating peptide and the cytostatic agent methotrexate (MTX). Two different peptides, YTA2 and YTA4, were designed and their intracellular delivery efficiency was characterized by fluorescence microscopy and quantified by fluorometry. MTX was conjugated to the transport peptides and the ability of the peptide-MTX conjugates to inhibit dihydrofolate reductase, the target enzyme of MTX, was found to be 15 and 20 times less potent than MTX. In addition, in vitro studies were performed in a drug resistant cell model using the 100-fold MTX resistant breast cancer cells MDA-MB-231. At a concentration of 1 microM, the peptide-MTX conjugates were shown to overcome MTX resistance and kill the cells more efficiently than MTX alone. Estimated EC50's were determined for MTX, MTX-YTA2 and YTA2 to be 18.5, 3.8 and 20 microM, respectively. In summary, cell-penetrating peptide conjugation of MTX is a new way of increasing delivery, and thereby, the potency of already well characterized therapeutic molecules into drug resistant tumour cells. PMID- 16376308 TI - Contribution of an unidentified sodium-dependent nucleoside transport system to the uptake and cytotoxicity of anthracycline in mouse M5076 ovarian sarcoma cells. AB - In the present study, we investigated whether an unidentified system for Na(+) dependent nucleoside transport is expressed by mouse M5076 ovarian sarcoma cells, besides concentrative nucleoside transporter 2 (CNT2(M)), and is involved in the uptake and cytotoxicity of anthracyclines. In a transport assay involving CNT2(M) transfectants, CNT2(M) was found to transport [(3)H]cytidine in a Na(+)-dependent manner, and 500 microM cytidine completely inhibited the Na(+)-dependent uptake of [(3)H]uridine via the transporter. In contrast, the Na(+)-dependent [(3)H]uridine uptake by M5076 cells decreased with 500 microM cytidine only to 70% of the control level. Furthermore, transfection of CNT2(M)-specific siRNAs into M5076 cells resulted in a reduction in the Na(+)-dependent uptake of [(3)H]uridine by only 23%, although the expression of CNT2(M) mRNA and Na(+) dependent uptake of [(3)H]cytidine disappeared in the cells. The uptake of pirarubicin (THP), an anthracycline, by M5076 cells requiring extracellular Na(+) was significantly inhibited by 500 microM uridine, but not 500 microM cytidine. The Na(+)-dependent and cytidine-insensitive uptake of [(3)H]uridine and the that of THP by M5076 cells significantly increased on cotreatment with both cholate and taurocholate, and the enhancement of THP uptake by the bile acids was reversed by cotreatment with 500 microM uridine. Furthermore, the cytotoxicity of THP and doxorubicin, which were previously reported to be taken up via the same transporter, toward M5076 cells was enhanced by cotreatment with both the bile acids. Therefore, it was indicated that an unidentified Na(+)-dependent transport system for nucleosides is expressed by M5076 cells, and contributes to the uptake and cytotoxicity of the anthracyclines. PMID- 16376309 TI - The effects of prostaglandin F2alpha treatment on peripheral-type benzodiazepine receptors in the ovary and uterus during pseudopregnancy of rats. AB - A previous study by us indicated that peripheral-type benzodiazepine receptor (PBR) density may be increased in the ovaries and uterus of pregnant rats (Weizman R, Dagan E, Snyder SH, Gavish M. Impact of pregnancy and lactation on GABAA receptor and central-type and peripheral-type benzodiazepine receptors. Brain Res 1997;752:7-14). In the present study, the effects of prostaglandin F2alpha (PGF2alpha) on PBR density in the ovary and uterus of pseudopregnant rats were assayed. Pseudopregnancy was induced on day 29 post-partum (PP) by s.c. injection of 50IU pregnant mare serum gonadotropin (PMSG) and 3 days later by s.c. injection of 20IU human chorionic gonadotropin (hCG). PBR ligand binding density was assayed with the specific PBR ligand [3H]PK 11195. A two-fold increase in ovarian PBR density was observed 2 days after hCG administration compared with vehicle control rats and this effect was maintained for 3 weeks. In the uterus, a three-fold increase in PBR density was observed and this increase was maintained for 1 week after hCG administration. Pseudopregnancy did not appear to affect renal PBR density or affinity. Treatment with PGF2alpha, which causes luteolysis, resulted in an approximately 50% reduction of PBR density in the ovaries of pseudopregnant rats at day 53 PP compared to pseudopregnant control rats. Treatment with indomethacin, which prevents the formation of PGF2alpha, caused the PBR density in the uterus of pseudopregnant rats at day 53 PP to be twice as high as in pseudopregnant control rats. All the above treatments did not affect the affinity of [3H]PK 11195 to ovarian and uterine PBR. These data suggest that PBR density in corpora lutea and uterus during pseudopregnancy is regulated by PGF2alpha. PMID- 16376310 TI - Region-, age-, and sex-specific effects of fetal diazepam exposure on the postnatal development of neurosteroids. AB - Fetal exposure to diazepam (DZ), a positive modulator of GABA(A) receptors and an agonist at mitochondrial benzodiazine receptors, induces long-term neural and behavioral effects. This study evaluated whether the early manipulation influenced the normal development of brain levels of neurosteroids or altered steroid action at GABA(A) receptors. Pregnant dams were injected over gestation days 14 through 20 with DZ (2.5 mg/kg) or the vehicle. Male and female offspring were analyzed at five postnatal ages. The levels of progesterone (P), dihydroprogesterone (DHP), 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha THP), testosterone (T), dihydrotestosterone, and 5alpha-androstan-3alpha,17beta diol were measured in the cerebral cortex and diencephalon. The results indicated that development of brain steroid levels and the impact of fetal DZ exposure were region- and sex-specific. Age-related changes in brain steroids did not mirror associated changes in circulating P and T. Age regulated the levels of all 3 progestins in the cerebral cortex, and fetal DZ exposure interacted with the development of P and DHP. The development of 3alpha,5alpha-THP in the cortex was markedly influenced by sex, with levels in males decreasing over postnatal development whereas they increased over postpubertal development in females. An adolescent surge in T levels was observed in male cortex and fetal DZ exposure prevented that surge. Steroid levels in the diencephalon were altered by age mainly in females, and DZ exposure had little effect in this region. The data support region-specific regulation of brain steroid synthesis. Only in the cerebral cortex are relevant mechanisms readily modifiable by fetal DZ exposure. However, neither sex nor fetal DZ exposure altered the response of GABA(A) receptors in adult cortex to neurosteroid. PMID- 16376311 TI - Theta bursts set up glutamatergic as well as GABA-ergic plasticity in neonatal rat hippocampal CA1 neurons. AB - gamma-Aminobutyric acid (GABA) is inhibitory in adult, but excitatory in neonatal, neurons. The switch from excitatory to inhibitory action is due to a negative shift in the equilibrium potential for the GABA(A) receptor-mediated postsynaptic current (E(GABA-PSC)). Here, we report that, in neonatal rat hippocampal CA1 neurons, presynaptic theta-burst activation induces not only a shift in E(GABA-PSC) towards that in adult neurons, but also a recruitment of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-receptor-mediated postsynaptic currents. PMID- 16376312 TI - Nuclear factor-kappa B nuclear translocation in the cochlea of mice following acoustic overstimulation. AB - There is increasing evidence to suggest that the expression of many molecules in the lateral wall of the cochlea plays an important role in noise-induced stress responses. In this study, activation of the nuclear transcription factor nuclear factor-kappa B (NF-kappaB) was investigated in the cochlea of mice treated with intense noise exposure (4 kHz, octave band, 124 dB, for 2 h). The present noise exposure led to remarkable auditory brainstem response threshold shifts and cochlear damage on surface preparations. To assess the effects of noise exposure on NF-kappaB/DNA binding activity in the cochlea, we prepared nuclear extracts from the cochlea at different time points after noise exposure and carried out an electrophoretic mobility shift assay using a probe specific to NF-kappaB. NF kappaB/DNA binding was significantly enhanced in the cochlea 2-6 h after noise exposure and returned to basal levels after 12 h. Supershift analysis using antibodies against p65 and p50 proteins, which are components of NF-kappaB, demonstrated that enhancement of NF-kappaB/DNA binding was at least in part due to nuclear translocation of p65. An immunohistochemical study also showed that nuclear translocation of both p65 and p50 was observed in the lateral wall after noise exposure and that there may be a possible close association between p65 and enhanced inducible nitric oxide synthase expression. These results suggest that NF-kappaB may have a detrimental role in the response to acoustic overstimulation in the cochlea of mice. PMID- 16376313 TI - Schwann cell: a source of neurotrophic activity on cortical glutamatergic neurons in culture. AB - Glial cells secrete numerous soluble molecules that enhance the development and the survival of different neuronal types cultured in vitro. Schwann cells (SC) play an important role as they are the source of different trophic substances and present a great neurotrophic activity. The aim of this study is to investigate the influence of postnatal SC on embryonic glutamatergic neurons. Co-cultures of SC from sciatic nerve of postnatal rats and neurons from rat embryonic cerebral cortex were successfully established, and cells were immunocytochemically characterized using mono and polyclonal antibodies as different glial and neuronal markers. Furthermore, some neuronal cultures were added with Nerve Growth Factor (NGF) and Insulin-like Growth Factor (IGF) to compare to co cultures. Our results show that SC promote an increase in the number of glutamatergic cortical neurons; moreover, these neurons present an evidence of dense axonal and dendritic outgrowth even when were fed with conditioned medium obtained from SC cultures. In conclusion, our data suggest that substances produced by SC exert a positive effect on central neuron survival and differentiation as indicated by processes of elongation and that this activity is mediated by soluble factors. Therefore, it is possible to consider the SC as a source of growth factors and might be suitable for the development of a neuroprotective effect in neurodegenerative disorders. PMID- 16376314 TI - The effect of interpolation and perceptual difficulty on the visual potentials evoked by illusory figures. AB - Completion is the process by which the brain unifies and segregates the parts of an incomplete form. It is qualified as amodal when the form is placed behind an obstacle and modal when the form is at the foreground and closed by illusory contours. The N1, and sometimes the N2, deflections of the visual evoked potentials are known to be larger for modal figures, such as the Kanizsa triangle, than for control figures. This result is generally linked to completion or illusory contours, but it could also be related to a third process: the interpolation of the form by connecting its separate parts. To test the influence of interpolation, a modal triangle, an amodal triangle, a figure with outlined inducers, and a no-triangle figure were randomly presented to 26 subjects. The N1 evoked by the three triangle figures were all larger than the N1 to the no triangle figure. These results suggest that the N1 amplitude is largely determined by the possibility of interpolating a form in the figure. The greatest N1 to the modal figure further suggests that interpolation may be increased by modal completion and decreased by the features that diminish the saliency of triangle in the amodal figure and the figure with outlined inducers. On the other hand, the largest N2 was evoked by the amodal figure. This effect may index processes activated in response to the great difficulty in perceiving the triangle in the amodal figure, a difficulty that is initially caused by a conflict of perceptions characterizing this figure. PMID- 16376315 TI - Rodent BDNF genes, novel promoters, novel splice variants, and regulation by cocaine. AB - Results from studies using molecular and genetic methods in humans and rodents suggest that brain-derived neurotrophic factor (BDNF) is involved in the behavioral effects of abused drugs, making understanding of its genomic structure and regulation of substantial interest. Recently, we have reported that the human BDNF gene contains seven upstream exons that can each be spliced independently to the major BDNF coding exon to form diverse bipartite BDNF transcripts. We also identified a novel "BDNFOS" gene that is transcribed to produce alternatively spliced natural antisense transcripts (NATs); its fifth exon overlaps with the protein coding exon VIII of human BDNF. To better understand BDNF's genomic structure and differential regulation, we now describe the rodent BDNF gene and transcripts. This gene includes six bipartite transcripts that are generated by six independently transcribed exons, each of which is spliced to a major coding exon and a tripartite transcript that is composed of two upstream exons and one coding exon. In addition, we found no evidence for antisense, opposite strand BDNFOS gene transcripts in mice or rats. The BDNF rodent splice variants display specific patterns of differential expression in different brain regions and peripheral tissues. Acute cocaine administration increased striatal expression of a specific BDNF4 splice variant by up to 5-fold. Interestingly, however, neither experimenter- nor self-administered chronic cocaine administration enhanced striatal BDNF expression. These data suggest a role of specific BDNF promoter regions and regulatory sequences in stimulant-induced alterations in BDNF expression, and in the alterations that changed BDNF expression is likely to confer in the brain. PMID- 16376316 TI - Electrophysiological evidence for a natural/artifactual dissociation. AB - The aim of this work was to test whether the differences usually found between the processing of visual stimuli corresponding to natural and artifactual domains reflect the different ways in which these domains are organized in the brain or are rather due to varying tasks demands. For this purpose, we designed two tasks in which subjects had to classify a series of line drawings. In one task (semantic categorization), the subjects were asked to categorize the stimuli as corresponding either to the natural or the artifactual class, and in the other (gender decision), the subjects had to decide if the names of the stimuli corresponded to either the masculine or the feminine gender. Event-related potentials (ERPs) and reactions times (RTs) were registered during the two tasks. We found both quantitative and topographical differences between ERPs elicited by natural stimuli and those by artifactual stimuli. In the 50- to 200-ms period, ERPs were more positive for the natural stimuli in the categorization task, but more positive for the artifactual stimuli in the gender decision task. In addition, natural stimuli elicited larger P600 and were associated with shorter RTs than artifactual stimuli in the categorization task. These results likely reflect differences concerning the relative difficulty of processing the stimuli of each domain in each task. In the N400 range, in contrast, there were differences between the two domains which were independent of task. In the two tasks, natural and artifactual stimuli elicited ERPs with a different scalp distribution: ERPs were more positive at posterior (parietal and temporal) sites for the natural stimuli and in the frontal areas for the artifactual ones. This finding is consistent with the hypothesis that semantic knowledge associated with natural and artifactual domains is represented in separate subsystems with presumably different anatomical bases. PMID- 16376317 TI - Differential expression of NMDA and AMPA/KA receptor subunits in the inferior olive of postnatal rats. AB - We have employed immunohistochemistry to determine the expression patterns of receptor subunits of N-methyl-d-aspartate (NMDA-NR1 and NR2A/B) and alpha-amino-3 hydroxy-5-methyl-isoxazole-4-propionic acid/kainic acid (AMPA/KA-GluR1, GluR2, GluR2/3, GluR4, and GluR5/6/7) in the inferior olive of postnatal rats up to adulthood. Immunoreactivity for distinct receptor subunits was predominantly localized in the soma and dendrites of neurons. Semi-quantification showed that the overall immunoreactivity in the inferior olive of adults was intense for GluR1, moderate for NR1 and NR2A/B, and low for GluR2, GluR2/3, GluR4, and GluR5/6/7. At P7, GluR1 was restricted to the dorsomedial cell column, subnucleus beta, principal nucleus and ventrolateral protrusion while the other subunits were found in all subnuclei of the inferior olive. The immunoreactivities for all glutamate receptor subunits ranged from low to moderate. As the rats matured, the immunoreactivity of GluR4 decreased after the second postnatal week, while those of the other subunits showed a general trend of increase, reaching adult level during the third postnatal week. Double immunofluorescence revealed that all NR1 containing neurons exhibited NR2A/B immunoreactivity, indicating that native NMDA receptors comprise of hetero-oligomeric combinations of NR1 and NR2A/B. Furthermore, co-localization of NMDA and AMPA/KA receptor subunits was demonstrated in individual neurons of the inferior olive. All NR1-containing neurons exhibited GluR1 immunoreactivity, and all NR2A/B-containing neurons showed GluR5/6/7 immunoreactivity. Our data suggest that NMDA and AMPA/KA receptors are involved in glutamate-mediated neurotransmission, contributing to synaptic plasticity and reorganization of circuitry in the inferior olive during postnatal development. PMID- 16376318 TI - Correlation of psycho-neuroendocrine-immune (PNI) gene expression with symptoms of acute infectious mononucleosis. AB - Acute infection is known to perturb psycho-neuroendocrine-immune (PNI) gene expression. Oligonucleotide microarrays were used to examine PNI gene expression in the peripheral blood of 13 subjects with infectious mononucleosis (IM). Novel peripheral blood gene expression activity was correlated with central-nervous system-mediated symptoms including fatigue and sleep disturbance. Of note, expression of the MADS box transcription enhancer factor 2 polypeptide C (MEF2C) gene, previously implicated in skeletal muscle myogenesis, correlated with symptoms of musculo-skeletal pain and fatigue. Expression of the hypocretin/orexin receptor HCRTR2, which has been implicated in narcolepsy, correlated with sleep disturbance. And, VACHT, the vesicular acetylcholine transporter, was highly correlated with neurocognitive disturbance. The expression of both HCRTR2 and MEF2C in the peripheral blood was validated by reverse transcription PCR. Thus, investigation of the PNI response in peripheral blood may provide novel insights into the complex pathophysiology of centrally mediated disease states. PMID- 16376322 TI - Atherosclerosis and proteinase activation. AB - Rapidly accumulating evidence points to the matrix metalloproteinases (MMPs) as major molecular mediators of arterial diseases. Findings from human pathological specimens, animals, and cell and molecular biology implicate matrix metalloproteinases in all stages of atherosclerosis including lesion initiation and progression and ultimately in plaque complication and triggering of thrombosis. The complex interactions within the proteolytic cascade allow multiple levels of control over these functions. This review weighs the evidence for the role of MMPs in arterial biology with particular reference to their activation in the atherosclerotic plaque. PMID- 16376323 TI - Direct effects of leptin on size and extracellular matrix components of human pediatric ventricular myocytes. AB - OBJECTIVE: There is a well-documented association between obesity and heart failure although the mechanistic basis for this correlation is unclear. Both extracellular matrix remodeling and left ventricular hypertrophy are well-defined components of remodeling in heart failure, and here we further investigate the role of leptin, the obese gene product, on these parameters. METHODS: We used primary human pediatric ventricular cardiomyocytes combined with gelatin zymography, quantitative PCR analysis, proline and leucine incorporation assays, and investigation of kinase activation by Western blotting. RESULTS: We show using gelatin zymography that leptin dose-dependently (0-60 nM) increased proteolytic activity at approximately 72 kDa. Accordingly, upon quantitative PCR analysis we found that leptin increased expression of matrix metalloproteinase-2 (MMP-2). Leptin also caused an increase in collagen type III and IV mRNA expression and a decrease in collagen type I mRNA expression. This was reflected in no significant change in total collagen synthesis, measured by [3H]proline incorporation, in response to leptin. A statistically significant increase in cell size, [3H]leucine incorporation, and expression of well-characterized markers of cardiac hypertrophy, namely cardiac alpha-actin and myosin light chain, were observed in response to leptin. We demonstrate activation of Janus activated kinase and mitogen-activated protein kinase pathways by leptin, and using pharmacological inhibitors we show that these signaling pathways play a role in mediating the effects of leptin. CONCLUSIONS: Our findings show that leptin regulates cell size, stimulates MMP-2 expression, and alters the profile, but not the total content, of collagen in human cardiomyocytes. This indicates the potential for altered leptin sensitivity to directly regulate cardiac remodeling in obesity. PMID- 16376324 TI - Cardiac mast cell regulation of matrix metalloproteinase-related ventricular remodeling in chronic pressure or volume overload. AB - The chronic elevation in ventricular wall stress secondary to ventricular volume or pressure overload leads to structural remodeling of the muscular, vascular and extracellular matrix components of the myocardium. While initially a compensatory response, the progressive hypertrophy and ventricular dilatation induced by this condition ultimately have a detrimental effect on ventricular function, resulting in heart failure. Fibrillar collagen provides the skeletal framework which interconnects the cardiomyocytes, thereby maintaining ventricular shape and size and contributing to tissue stiffness. Accordingly, these myocardial collagen fibers must be disrupted for ventricular dilatation, sphericalization and wall thinning to occur. The presence of an abundant, latent matrix metalloproteinase (MMP) population which coexists with myocardial fibrillar collagen has been documented. Thus, the potential for collagen degradation to exceed synthesis exists should there be significant activation of this latent MMP system. Mast cells are known to store and release a variety of biologically active mediators including TNF-alpha and proteases such as tryptase and chymase, which can induce MMP activation. Increased cardiac mast cell density has been implicated in the pathophysiology of human end-stage cardiomyopathy and experimental myocardial infarction, hypertension and chronic volume overload secondary to mitral regurgitation and aorto-caval fistula. The potential role of cardiac mast cells in activating MMPs, which then results in fibrillar collagen degradation and adverse myocardial remodeling secondary to chronic volume and pressure overload will be the subject of this review. PMID- 16376325 TI - Changes in cardiac lipid metabolism during sepsis: the essential role of very low density lipoprotein receptors. AB - OBJECTIVE: Sepsis accompanies myocardial dysfunction and dynamic alterations of cardiac metabolism. We have recently demonstrated that the very low-density lipoprotein receptor (VLDL-R), which is abundantly expressed in the heart, plays a key role in energy metabolism of the fasting heart. However, little is known about the function and regulation of the VLDL-R during sepsis. In the present study, we explored lipid accumulation and VLDL-R expression in the lipopolysaccharide (LPS)-stimulated heart in vivo and regulation of VLDL-R expression in vitro. METHODS AND RESULTS: Electron microscopy and immunohistochemistry demonstrated that LPS significantly decreased both lipid accumulation and VLDL-R expression in the hearts of fasting mice. Treatment with LPS also downregulated VLDL-R in rat neonatal cardiac myocytes, and this downregulation was completely reversed by interleukin (IL)-1beta receptor antagonist. IL-1beta downregulated the expression of VLDL-R in a time- and dose dependent manner and markedly reduced the uptake of DiI-labeled beta-VLDL but not DiI-labeled low-density lipoprotein (LDL). Use of specific pharmacologic inhibitors and short interference RNA revealed that Hsp90 was required for IL 1beta to downregulate VLDL-R expression. CONCLUSIONS: These findings suggest that IL-1beta is a principle mediator of changes in cardiac lipid and energy metabolism during sepsis through the downregulation of myocardial VLDL-R expression. PMID- 16376326 TI - Alterations of the preproenkephalin system in cardiac hypertrophy and its role in atrioventricular conduction. AB - OBJECTIVE: The goal of this study was to investigate alterations of the endogenous opioid system in cardiac hypertrophy, to elucidate mechanisms of preproenkephalin (ppENK) gene expression, and to assess effects of endogenous opioids on myocardial contractility and atrioventricular conduction. METHODS: Hypertrophy was induced by ligation of a renal artery (2K1C) or chronic isoprenaline infusion (ISO). ppENK and opioid receptor (mu-, delta-, kappa-OR) mRNA expression was quantified by Northern blot and quantitative RT-PCR, respectively. Isolated cardiac myocytes and non-myocytes from neonatal rat heart were used for cell culture experiments. RESULTS: Overall expression of OR in the heart was markedly lower than in brain tissue, with kappa-OR being the most abundant isoform in the heart. We did not observe differences in kappa-OR expression in ventricular and atrial myocardium. In contrast, delta-OR expression was markedly higher in atria than in ventricles. Mu-OR expression in the heart was below the detection limit of the developed qRT-PCR assay. In left ventricular myocardium, ppENK mRNA levels were significantly increased in 2K1C rats but decreased in ISO rats. Cell culture experiments from neonatal rat hearts revealed that myocytes and non-myocytes express ppENK mRNA. In these cells, receptor dependent and -independent stimulation of the beta-adrenergic signalling pathway caused an increase in ppENK mRNA. Furthermore, inactivation of inhibitory G proteins by pertussis toxin increased basal and noradrenaline-stimulated ppENK mRNA expression. The physiological significance of myocardial opioids was investigated in isolated perfused rat hearts. Opioid receptor antagonists (nor BNI, naltrindol) and the enkephalinase inhibitor CPL had no effect on contractility but significantly altered atrioventricular conduction. CONCLUSION: These observations suggest that the cardiac opioid system is activated in cardiac hypertrophy. Pressure overload and stimulation of the beta-adrenergic signalling pathway have been identified as a possible mechanism leading to increased ppENK expression, which may contribute to opioid system activation. Finally, endogenous opioids modulate the dromotropic response to catecholamine stimulation. The latter finding raises the possibility that endogenous opioids may contribute to the pathogenesis of arrhythmias. PMID- 16376327 TI - Autologous myoblast transplantation after myocardial infarction increases the inducibility of ventricular arrhythmias. AB - BACKGROUND: Small scale clinical trials suggested the feasibility and the efficacy of autologous myoblast transplantation to improve ventricular function after myocardial infarction. However, these trials were hampered by unexpected episodes of life-threatening ventricular tachyarrhythmias (VT). We investigated cardiac electrical stability after myoblast transplantation to the myocardium. METHODS AND RESULTS: Seven days after coronary ligation, Wistar rats were randomized into 3 groups: a control group receiving no further treatment, a vehicle group injected with culture medium into the infarcted myocardium, and a myoblast group injected with autologous myoblasts. Holter monitoring did not discriminate the myoblast from the vehicle groups. Programmed Electrical Stimulation (PES) was performed to evaluate further a cardiac substrate for arrhythmia susceptibility. The occurrence of sustained VT during PES was similar in control and vehicle groups (5/17 and 4/19 rats, respectively; p=0.50). In contrast, 13/20 rats (65%) from the myoblast group showed at least one episode of sustained VT during PES (p<0.05 and p<0.005 versus control and vehicle groups). As a further control group, rats injected with autologous bone marrow mononuclear cells into the infarcted myocardium did not show increased susceptibility to PES. CONCLUSIONS: In an infarcted rat model, myoblast transplantation but not bone marrow mononuclear cells or myocardial injection per se induces electrical ventricular instability. Because ventricular arrhythmias are life-threatening disorders, we suggest that such preclinical evaluation should be conducted for any new source of cells to be injected into the myocardium. PMID- 16376328 TI - Measurement of testosterone and its sub-fractions in Canada. AB - Interest in measuring bioactive testosterone in aging males has increased considerably in the last 5 years in Canada. Emerging andropause clinics have submerged our laboratories with requests for bioavailable testosterone (BAT) testing in replacement or addition to the traditional total testosterone (TT) and direct free testosterone (FT) assays. Beginning with a brief explanation of the bioavailability concept of Pardridge, this review examines the technical characteristics of various approaches currently available to measure TT and its sub-fractions. First, limitations in the measurement of TT, SHBG, and particularly direct (analog) FT assays are extracted from the scientific literature and recent external and internal QC reports. It is concluded that the free direct T assay is useless in the clinical context of andropause. The impact of the observed limitations of TT and SHBG measurements on calculated FT and BAT or BAT obtained by precipitation with ammonium sulfate is then discussed. A comparative evaluation of the advantages and disadvantages of calculated FT or BAT vs. precipitated BAT is presented before concluding that doing a TT as a first line test remains overall the most cost-effective measurement in the diagnosis of hypogonadism in males, and that this sole determination will be sufficient in over 75% of the cases. PMID- 16376329 TI - Regulation of neuronal lineage decisions by the HES-related bHLH protein REF-1. AB - Members of the HES subfamily of bHLH proteins play crucial roles in neural patterning via repression of neurogenesis. In C. elegans, loss-of-function mutations in ref-1, a distant nematode-specific member of this subfamily, were previously shown to cause ectopic neurogenesis from postembryonic lineages. However, while the vast majority of the nervous system in C. elegans is generated embryonically, the role of REF-1 in regulating these neural lineage decisions is unknown. Here, we show that mutations in ref-1 result in the generation of multiple ectopic neuron types derived from an embryonic neuroblast. In wild-type animals, neurons derived from this sublineage are present in a left/right symmetrical manner. However, in ref-1 mutants, while the ectopically generated neurons exhibit gene expression profiles characteristic of neurons on the left, they are present only on the right side. REF-1 functions in a Notch-independent manner to regulate this ectopic lineage decision. We also demonstrate that loss of REF-1 function results in defective differentiation of an embryonically generated serotonergic neuron type. These results indicate that REF-1 functions in both Notch-dependent and independent pathways to regulate multiple developmental decisions in different neuronal sublineages. PMID- 16376330 TI - Antiangiogenesis in cancer therapy--endostatin and its mechanisms of action. AB - The first angiogenesis inhibitors for cancer have now been approved by the F.D.A. in the U.S. and in 28 other countries, including China. The majority of these are monotherapies that block VEGF. However, mutant tumor cells may over time produce redundant angiogenic factors. Therefore, for long-term use in cancer, combinations of angiogenesis inhibitors or broad spectrum angiogenesis inhibitors will be needed. The two most broad spectrum and least toxic angiogenesis inhibitors are Caplostatin and endostatin. Endostatin inhibits 65 different tumor types and modifies 12% of the human genome to downregulate pathological angiogenesis without side-effects. The recent discovery that small increases in circulating endostatin can suppress tumor growth and that orally available small molecules can increase endostatin in the plasma suggests the possible development of a new pharmaceutical field. PMID- 16376331 TI - Identification of cytoplasmic p53 protein in corneal epithelium of vertebrates. AB - The tumour suppressor gene p53 encodes a phosphoprotein involved in the control of cell growth. It's expression and function have been documented in malignancy, apoptosis and the aging processes. Recently, p53 expression has been demonstrated in normal murine tissues, including whole eye. Currently, we intend to map and to characterize p53 expression in the normal cornea across different species. To do this, eyes of animals were enucleated after sacrifice by CO(2) narcosis and then p53 expression in whole eyes (cornea) was mapped by indirect immunohistochemical staining techniques using the anti-p53 monoclonal antibodies PAb 248, PAb 421 and PAb 240 (alternatively called mAb 248, mAb 421 and mAb 240, respectively). Additionally, eyes were freshly dissected to separate the corneas, for quantitating p53 expression, using Western blot analysis. We found strong cytoplasmic p53 expression in the corneal epithelium of various vertebrate species by immunohistochemistry and by Western analysis. High levels of cytoplasmic p53 protein were normally found in normal corneal epithelium of various vertebrate species. Hence, these data may indicate that p53 may have a new evolutionary significant function in the eye. PMID- 16376332 TI - Trypanosoma cruzi: sensitivity of the polymerase chain reaction for detecting the parasite in the blood of mice infected with different clonal genotypes. AB - The polymerase chain reaction showed high sensitivity for detecting Trypanosoma cruzi in the blood of mice, independent of clonal genotype (19, 20-T. cruzi I; 32, 39-T. cruzi II) or phase of the infection (acute or chronic). PMID- 16376333 TI - Leishmania major: species specific delayed hypersensitivity reaction induced by exogenous secreted antigen in the guinea pig. AB - The cellular response to Leishmania major (L. major) is usually evaluated in vivo by the delayed-type-hypersensitivity (DTH) test using leishmanin. Leishmanin can give false-positive reactions in areas where there is a background of leishmaniasis. In a previous study, it was shown that a 56 kDa antigen purified from promastigote and culture supernatant of L. major induce strong DTH reactions in sensitized guinea pigs. In this study, the species-specificity of this antigen was further investigated. Three groups of guinea pigs were sensitized with L. major, L. tropica, and L. infantum and both flanks of sensitized animal were injected intradermally with purified 56 kDa antigen or soluble leishmania antigen (SLA). The extent of indurations were measured after 24, 48, and 72 h. In animals which were sensitized with three species of leishmania, only those immunized with L. major showed skin reactions to purified antigen by an increase in skin thickness. Since complex antigen mixtures such as SLA and leishmanin show cross reactivity and can be non-specific, the result obtained here suggest that 56 kDa antigen may be a useful diagnostic tool for species specific diagnosis in field studies of leishmaniasis. PMID- 16376334 TI - ABC lipid transporters: extruders, flippases, or flopless activators? AB - Many mammalian ABC transporters move membrane lipids to acceptor lipid assemblies in the extracellular aqueous milieu. Because the desorption from the membrane costs more energy than provided by two ATPs, the transporter probably only translocates the lipid to a partially hydrophilic site on its extracellular face. From this high-energy site, the lipid may efficiently move to the acceptor, which ideally is bound to the transporter, or, in the absence of an acceptor, fall back into the membrane. If the lipid originated from the cytosolic membrane surface, this represents lipid flop and is probably a side activity of the transporters. PMID- 16376335 TI - Phosphorylation and concomitant structural changes in human 2-Cys peroxiredoxin isotype I differentially regulate its peroxidase and molecular chaperone functions. AB - The H2O2-catabolizing peroxidase activity of human peroxiredoxin I (hPrxI) was previously shown to be regulated by phosphorylation of Thr90. Here, we show that hPrxI forms multiple oligomers with distinct secondary structures. HPrxI is a dual function protein, since it can behave either as a peroxidase or as a molecular chaperone. The effects of phosphorylation of hPrxI on its protein structure and dual functions were determined using site-directed mutagenesis, in which the phosphorylation site was substituted with aspartate to mimic the phosphorylated status of the protein (T90D-hPrxI). Phosphorylation of the protein induces significant changes in its protein structure from low molecular weight (MW) protein species to high MW protein complexes as well as its dual functions. In contrast to the wild type (WT)- and T90A-hPrxI, the T90D-hPrxI exhibited a markedly reduced peroxidase activity, but showed about sixfold higher chaperone activity than WT-hPrxI. PMID- 16376336 TI - Crystal structure of the PB1 domain of NBR1. AB - The scaffold protein NBR1 is involved in signal transmission downstream of the serine/protein kinase from the giant muscle protein titin. Its N-terminal Phox and Bem1p (PB1) domain plays a critical role in mediating protein-protein interactions with both titin kinase and with another scaffold protein, p62. We have determined the crystal structure of the PB1 domain of NBR1 at 1.55A resolution. It reveals a type-A PB1 domain with two negatively charged residue clusters. We provide a structural perspective on the involvement of NBR1 in the titin kinase signalling pathway. PMID- 16376337 TI - NF-kappaB activation is required for cisplatin-induced apoptosis in head and neck squamous carcinoma cells. AB - This study demonstrates a requirement for NF-kappaB activation in cis diamminedichloroplatinum (cisplatin)-induced apoptosis in human head and neck squamous cell carcinoma (HNSCC) cell lines. This conclusion was supported by the following observations: cisplatin induced IkappaBalpha degradation and NF-kappaB dependent transcriptional activation prior to cell death; pyrrolidine dithiocarbamate (PDTC), a chemical inhibitor of NF-kappaB activation, prevented apoptosis; lactacystin, an inhibitor of IkappaBalpha degradation, also prevented apoptosis; and finally, the expression of a super-repressor mutant IkappaBalpha blocked apoptosis. The expression of tumor necrosis factor alpha (TNFalpha) was promoted by cisplatin treatment and was suppressed by PDTC treatment. In addition, a neutralizing antibody against TNFalpha protected cells from cisplatin induced apoptosis. These findings suggest that NF-kappaB activation is required for cisplatin-induced apoptosis and TNFalpha may play an important role in NF kappaB-mediated apoptosis in cisplatin-treated HNSCC cell lines. PMID- 16376338 TI - Protein kinase A phosphorylates and regulates dimerization of 14-3-3 epsilon. AB - Recognition of phosphorylated serine/threonine-containing motifs by 14-3-3 depends on the dimerization of 14-3-3. However, the molecular cues that control 14-3-3 dimerization are not well understood. In order to identify proteins that control 14-3-3 dimerization, we analyzed proteins that have effects on 14-3-3 dimerization and report that protein kinase A (PKA) phosphorylates 14-3-3zeta at a specific residue (Ser58). Phosphorylation by PKA leads to modulation of 14-3 3zeta dimerization and affect its interaction with partner proteins. Substitution of Ser58 to Ala completely abolished phosphorylation of 14-3-3zeta by PKA. A phospho-mimic mutant of 14-3-3zeta, Ser58 to Glu substitution, failed to form homodimers, showed reduced interaction with 14-3-3epsilon and p53, and could not enhance transcriptional activity of p53. Moreover, activation of PKA decreases and inhibition of PKA increases the dimerization of 14-3-3zeta and the functional interaction of 14-3-3zeta with p53. Therefore, our results suggest that PKA is a new member of protein kinases that can phosphorylate and impair the function of 14-3-3. PMID- 16376339 TI - The mechanisms of regulation of Hdm2 protein level by serum growth factors. AB - Cell cycle progression in response to serum growth factors is dependent on the expression of functional Hdm2 (Mdm2), which inhibits p53-dependent transcription of anti-proliferative genes. In a well characterised non-transformed human fibroblast model, growth factors induce the expression of Hdm2 with rapid kinetics. Here we dissect the mechanistic basis for this critical response. In contrast to previous studies in which components of the growth factor signalling pathways were overexpressed, hdm2 mRNA expression is not induced with immediate early kinetics in these cells. Rather, the elevated Hdm2 protein levels which follow growth factor stimulation are primarily a consequence of phosphatidylinositol-3 kinase-dependent stabilisation of the Hdm2 protein combined with a global increase in protein synthesis. PMID- 16376340 TI - Novel bioactive parathyroid hormone and related peptides in teleost fish. AB - We report the identification, gene expression and biological activity of two parathyroid hormones (PTH; PTHA and PTHB), two PTH-related peptides (PTHrP; PTHrPA and PTHrPB) and a PTH-like ligand (PTH-L) with hybrid characteristics in puffer fishes (Takifugu rubripes and Tetraodon fluviatilis). Experimental data are consistent with PTH-L and PTHrPA having calciotropic activities equivalent, respectively, to tetrapod PTH and PTHrP. We hypothesise on the basis of phylogenetic and functional analysis that PTH-L could be a fish relic of an ancestral PTH/PTHrP gene. PMID- 16376341 TI - Dehydroepiandrosterone increases beta-cell mass and improves the glucose-induced insulin secretion by pancreatic islets from aged rats. AB - The effect of dehydroepiandrosterone (DHEA) on pancreatic islet function of aged rats, an animal model with impaired glucose-induced insulin secretion, was investigated. The following parameters were examined: morphological analysis of endocrine pancreata by immunohistochemistry; protein levels of insulin receptor, IRS-1, IRS-2, PI 3-kinase, Akt-1, and Akt-2; and static insulin secretion in isolated pancreatic islets. Pancreatic islets from DHEA-treated rats showed an increased beta-cell mass accompanied by increased Akt-1 protein level but reduced IR, IRS-1, and IRS-2 protein levels and enhanced glucose-stimulated insulin secretion. The present results suggest that DHEA may be a promising drug to prevent diabetes during aging. PMID- 16376342 TI - Tea polyphenol epigallocatechin 3-gallate impedes the anti-apoptotic effects of low-grade repetitive stress through inhibition of Akt and NFkappaB survival pathways. AB - V79 Chinese Hamster lung fibroblasts were subjected to repetitive low-grade stress through multiple exposures to 30 microM H2O2 in culture for 4 weeks. Akt/protein kinase B became phosphorylated at serine473 and threonine308 during this period of repetitive stress. Concurrent exposure of the cells to LY294002 (5 microM), a phosphoinositide-3 kinase inhibitor or 4.5 microM epigallocatechin 3 gallate (EGCG), a tea polyphenol almost completely blocked Akt activation by repetitive stress. Phosphorylation of I kappa B kinase (IKK) and transcriptional activity driven by nuclear factor kappa B (NFkappaB) were significantly enhanced by repetitive oxidative stress. These increases were largely abolished by simultaneous exposure to EGCG. The repetitively stressed cells demonstrated a significant resistance to apoptosis by subsequent acute stress in the form of ultraviolet radiation at 5 J/m2 or H2O2 (7.5 mM). The resistance to apoptosis conferred by repetitive stress was drastically reduced (>80%) by constant exposure to EGCG during the stress period while the presence of LY294002 or the NFkappaB inhibitor SN50 brought about a relatively moderate effect (about 50 65%). Our data indicate that activation of Akt and NFkappaB pro-survival pathways by repetitive low-grade stress results in a strong inhibition of the normal apoptotic response after subsequent acute stress. The tea polyphenol EGCG impedes the activation of both Akt and NFkappaB by repetitive stress and as a result preserves the normal apoptotic response during subsequent acute stress. PMID- 16376343 TI - Structural insight into binding of Staphylococcus aureus to human fibronectin. AB - Staphylococcus aureus possesses cell-wall attached proteins that bind the human protein fibronectin (Fn). An intermodule interface between the 4F1 and 5F1 modules in the N-terminal domain of Fn is maintained on bacterial peptide binding but there is a small change in the intermodule orientation and alignment of beta strands that are predicted to bind the peptide. The module pair is elongated, as in the unbound state. Combined with evidence that residues in both 4F1 and 5F1 are directly involved in peptide binding, this observation supports the hypothesis that, when bound to intact Fn, the bacterial protein adopts an unusual, highly extended conformation. PMID- 16376344 TI - FCH/Cdc15 domain determines distinct subcellular localization of NOSTRIN. AB - NOSTRIN, an NO synthase binding protein, belongs to the PCH family of proteins, exposing a typical domain structure. While its SH3 domain and the C-terminal coiled-coil region cc2 have been studied earlier, the function of the N-terminal half comprising a Cdc15 domain with an FCH (Fes/CIP homology) region followed by a coiled-coil stretch cc1 is unknown. Here, we show that the FCH region is necessary and sufficient for membrane association of NOSTRIN, whereas the Cdc15 domain further specifies subcellular distribution of the protein. Thus, the FCH region and the Cdc15 domain fulfill complementary functions in subcellular targeting of NOSTRIN. PMID- 16376345 TI - Diagnostic performance of serial haematocrit measurements in identifying major injury in adult trauma patients. AB - Determination of occult haemorrhage is an essential part of trauma assessment. We evaluated the diagnostic utility of decreasing haematocrit (DeltaHct) in detecting major injury. Additionally, we tested the correlation between the volume of infused intravenous fluid (IVF) and DeltaHct. METHODS: Prospective observational study at a level one trauma centre. INCLUSION CRITERIA: Patients with suspected major injury. exclusion criteria: Patients who received blood transfusion in the first 4 h, and those who deceased or were transferred to other units before the completion of the observation period (4 h). We measured IVF and DeltaHct at 4 h after triage. We classified patients as having minor or major injury on the basis of injury severity score > or =15. Receiver Operating Characteristic (ROC) curve was used to test the diagnostic performance of DeltaHct in identifying major injury. We tested the operating characteristics of DeltaHct cut-off values of 5 and 10 in detecting major injury. We also measured the correlation of IVF and DeltaHct in a subgroup of patients with low potential for blood loss (ISS<3) to account for possibility of haemodilution. RESULTS: Four hundred and ninety-four patients (convenience sample) were enrolled (age 36+/-17 years, 82% male, 57% blunt trauma). Sixty-three patients (13%) had major injury. The area under the ROC curve for DeltaHct was not significantly different from the unity line (p=0.20). DeltaHct-4 h>5 points had a sensitivity of 40% (95% CI, 29-52%), specificity of 94% (95% CI, 92-96%), likelihood ratio for a positive test (LR+) of 7.1 (95% CI, 4.4-11.7), and likelihood ratio for a negative test ( LR) of 0.64 (95% CI, 0.52-0.78) in identifying major trauma. DeltaHct-4 h>10 points had sensitivity of 16% (95% CI, 9-27%), specificity of 95% (95% CI, 92 0.96%), +LR of 3.0 (95% CI, 1.5-5.9), and -LR of 0.89 (95% CI, 0.80-0.99). In our subgroup analysis, we detected no significant correlation (p=0.09) between the IVF and DeltaHct-4 h. CONCLUSIONS: DeltaHct-4 h>5 or 10 points is suggestive of major injury (high specificity and +LR). However, the failure to drop the Hct cannot be used to rule out major injury (low sensitivity and -LR). PMID- 16376346 TI - Live births, cesarean sections and the development of menstrual abnormalities. AB - OBJECTIVES: To examine the relationship between number of livebirths and cesarean sections on the development of menstrual abnormalities. METHODS: The effect of number of livebirths and cesarean sections on menstrual abnormalities in the late reproductive period was evaluated using data from the Harvard Study of Moods and Cycles. RESULTS: There were 298 women with no livebirths and 606 women with livebirths of which 184 had a history of cesarean section. Among all women, increasing number of livebirths was associated with an increasing trend in reported heavy menses, prolonged flow >6 days, and a decreasing trend in reported dysmenorrhea. This effect was amplified among women with a history of cesarean section. CONCLUSIONS: Increasing number of livebirths is associated with heavy and prolonged menstrual flow during the late reproductive period. This association is particularly stronger in women with a history of cesarean section. PMID- 16376347 TI - Work situation of registered nurses in municipal elderly care in Sweden: a questionnaire survey. AB - BACKGROUND: Organizational changes have occurred in municipal elderly care in Sweden during the past decades. The 'Adel' reform transferred responsibility for the care of older persons from the county councils to the municipalities. Furthermore, the specialisation in dementia care divided elderly care into two groups: dementia and general care. This change has had a significant impact on the work situation of registered nurses (RNs). AIM: The main focus was to describe RNs' work situation and their characteristics in municipal elderly care. Another aim was to compare RNs working solely in dementia care with those working in general care of older persons with diverse diagnoses. DESIGN: A non experimental, descriptive design with a survey research approach was used. SETTINGS: Sixty special housing units with underlying units including those offering daytime activities in a large city in the middle of Sweden. PARTICIPANTS: The number of participating RNs was a total of 213, with a response rate of 62.3%. Of the 213 RNs, 95 (44.6%) worked in dementia care, and 118 (55.4%) in general care. METHOD: A questionnaire survey. RESULTS: The results indicated high levels of time pressure in both groups. Greater knowledge and greater emotional and conflicting demands were found in dementia care. The majority perceived a greater opportunity to plan and perform daily work tasks than to influence the work situation in a wider context. Support at work was perceived as generally high from management and fellow workers and higher in dementia care. CONCLUSION: It is important to decrease RNs' time pressure and increase their influence on decisions made at work. PMID- 16376348 TI - Is the introduction of a student-centred, problem-based curriculum associated with improvements in student nurse well-being and performance? An observational study of effect. AB - AIMS: To explore the impact of curriculum redesign and innovation on student well being and performance, including essay and examination marks and sickness absence. BACKGROUND: While the emotional impact of preparing to be a health professional can be reduced by helping students to adapt, the positive effect of curriculum innovation and redesign is rarely evaluated. DESIGN, SAMPLE AND METHODS: Student nurse well-being and performance was compared at weeks 24/25 and 40/50 following course entry between comparable independent cohorts of students undertaking a traditional programme (N=406, 83% return rate) with those on an innovative, student-centred, problem-based educational programme (N=447, 79% return rate). The setting was a School of Nursing and Midwifery in the North-East of Scotland. Measures included stress and mental health outcomes and measures of performance including academic marks and sickness absence. RESULTS: At week 25 into the course students on the innovative course had fewer academic, clinical and personal worries than students in the previous more traditional programme and were more likely to report using adaptive direct, problem-solving coping at week 50. While students on the innovative course reported less distress in their first year of the course, they scored less well on comparable essay assignments and had reliably greater sickness absence totals than those educated by traditional methods. CONCLUSION: In this setting, curriculum innovation was associated with positive changes in student well-being but not on performance. PMID- 16376349 TI - Unconfined compression of white matter. AB - The porous properties of brain tissue are important for understanding normal and abnormal cerebrospinal fluid flow in the brain. In this study, a poroviscoelastic model was fitted to the stress relaxation response of white matter in unconfined compression performed under a range of low strain rates. A set of experiments was also performed on the tissue samples using a no-slip boundary condition. Results from these experiments demonstrated that the rheological response of the white matter is primarily governed by the intrinsic viscoelastic properties of the solid phase. The permeability of white matter was found to be of the order of 10( 12) m4/Ns. PMID- 16376350 TI - Dynamic dorsoventral stiffness assessment of the ovine lumbar spine. AB - Posteroanterior spinal stiffness assessments are common in the evaluating patients with low back pain. The purpose of this study was to determine the effects of mechanical excitation frequency on dynamic lumbar spine stiffness. A computer-controlled voice coil actuator equipped with a load cell and LVDT was used to deliver an oscillatory dorsoventral (DV) mechanical force to the L3 spinous process of 15 adolescent Merino sheep. DV forces (48 N peak, approximately 10% body weight) were randomly applied at periodic excitation frequencies of 2.0, 6.0, 11.7 and a 0.5-19.7 Hz sweep. Force and displacement were recorded over a 13-22 s time interval. The in vivo DV stiffness of the ovine spine was frequency dependent and varied 3.7-fold over the 0.5-19.7 Hz mechanical excitation frequency range. Minimum and maximum DV stiffness (force/displacement) were 3.86+/-0.38 and 14.1+/-9.95 N/mm at 4.0 and 19.7 Hz, respectively. Stiffness values based on the swept-sine measurements were not significantly different from corresponding periodic oscillations (2.0 and 6.0 Hz). The mean coefficient of variation in the swept-sine DV dynamic stiffness assessment method was 15%, which was similar to the periodic oscillation method (10-16%). The results indicate that changes in mechanical excitation frequency and animal body mass modulate DV spinal stiffness. PMID- 16376351 TI - Accuracy and repeatability of an optical motion analysis system for measuring small deformations of biological tissues. AB - Optical motion analysis techniques have been widely used in biomechanics for measuring large-scale motions such as gait, but have not yet been significantly explored for measuring smaller movements such as the tooth displacements under load. In principle, very accurate measurements could be possible and this could provide a valuable tool in many engineering applications. The aim of this study was to evaluate accuracy and repeatability of the Qualisys ProReflex-MCU120 system when measuring small displacements, as a step towards measuring tooth displacements to characterise the properties of the periodontal ligament. Accuracy and repeatability of the system was evaluated using a wedge comparator with a resolution of 0.25 microm to provide measured marker displacements in three orthogonal directions. The marker was moved in ten steps in each direction, for each of seven step sizes (0.5, 1, 2, 3, 5, 10, and 20 microm), repeated five times. Spherical and diamond markers were tested. The system accuracy (i.e. percentage of maximum absolute error in range/measurement range), in the 20-200 microm ranges, was +/-1.17%, +/-1.67% and +/-1.31% for the diamond marker in x, y and z directions, while the system accuracy for the spherical marker was +/ 1.81%, +/-2.37% and +/-1.39%. The system repeatability (i.e. maximum standard deviation in the measurement range) measured under the different days, light intensity and temperatures for five times, carried out step up and then step down measurements for the same step size, was +/-1.7, +/-2.3 and +/-1.9 microm for the diamond marker, and +/-2.6, +/-3.9 and +/-1.9 microm for the spherical marker in x, y and z directions, respectively. These results demonstrate that the system suffices accuracy for measuring tooth displacements and could potentially be useful in many other applications. PMID- 16376352 TI - Development of a locking femur nail for mice. AB - We herein report on a novel locking intramedullary nail system in a murine closed femur fracture model. The nail system consists of a modified 24-gauge injection needle and a 0.1-mm-diameter tungsten guide wire. Rotation stability was accomplished by flattening the proximal and distal end of the needle. Torsional mechanical testing of the implants in osteotomized cadaveric femora revealed a superiority of the locking nail (3.9+/-1.0 degrees rotation at a torque of 0.9 Nmm, n=10) compared to the unmodified injection needle (conventional nail; 52.4+/ 3.2 degrees, n=10, p<0.05). None of the implants, however, achieved the rotation stability of unfractured femora (0.3+/-0.5 degrees, n=10). In a second step, we tested the feasibility of the in vivo application of the locking nail to stabilize a closed femoral midshaft fracture in C57BL/6 mice. Of interest, none of the 10 animals showed a dislocation of the locking nail over a 5-week period, while 3 of 4 animals with conventional nail fracture stabilization showed a significant pin dislocation within the first 3 days (p<0.05). Mechanical testing after 5-weeks stabilization with the locking nail revealed an appropriate bone healing with a torque at failure of 71.6+/-3.4% and a peak rotation before failure of 68.4+/-5.3% relative to the unfractured contralateral femur. With the advantage that closed fractures can be fixed with rotation stability, the herein introduced model may represent an ideal tool to study bone healing in transgenic and knockout mice. PMID- 16376353 TI - Finger joint coordination during tapping. AB - We investigated finger joint coordination during tapping by characterizing joint kinematics and torques in terms of muscle activation patterns and energy profiles. Six subjects tapped with their index finger on a computer keyswitch as if they were typing on the middle row of a keyboard. Fingertip force, keyswitch position, kinematics of the metacarpophalangeal (MCP) and the proximal and distal interphalangeal (IP) joints, and intramuscular electromyography of intrinsic and extrinsic finger muscles were measured simultaneously. Finger joint torques were calculated based on a closed-form Newton-Euler inverse dynamic model of the finger. During the keystroke, the MCP joint flexed and the IP joints extended before and throughout the loading phase of the contact period, creating a closing reciprocal motion of the finger joints. As the finger lifted, the MCP joint extended and the interphalangeal (IP) joints flexed, creating an opening reciprocal motion. Intrinsic finger muscle and extrinsic flexor activities both began after the initiation of the downward finger movement. The intrinsic finger muscle activity preceded both the IP joint extension and the onset of extrinsic muscle activity. Only extrinsic extensor activity was present as the finger was lifted. While both potential energy and kinetic energy are present and large enough to overcome the work necessary to press the keyswitch, the motor control strategies utilize the muscle forces and joint torques to ensure a successful keystroke. PMID- 16376355 TI - Ultrahigh pressure liquid chromatography using elevated temperature. AB - Fast liquid chromatographic (LC) methods are important for a variety of applications. Reducing the particle diameter (d(p)) is the most effective way to achieve fast separations while preserving high efficiency. Since the pressure drop along a packed column is inversely proportional to the square of the particle size, when columns packed with small particles (<2 microm) are used, ultrahigh pressures (>689 bar) must be applied to overcome the resistance to mobile phase flow. Elevating the column temperature can significantly reduce the mobile phase viscosity, allowing operation at higher flow rate for the same pressure. It also leads to a decrease in retention factor. The advantage of using elevated temperatures in LC is the ability to significantly shorten separation time with minimal loss in column efficiency. Therefore, combining elevated temperature with ultrahigh pressure facilitates fast and efficient separations. In this study, C6-modified 1.0 microm nonporous silica particles were used to demonstrate fast separations using a temperature of 80 degrees C and a pressure of 2413 bar. Selected separations were completed in 30 s with efficiencies as high as 220,000 plates m(-1). PMID- 16376354 TI - Computational and experimental models of the human torso for non-penetrating ballistic impact. AB - Both computational finite element and experimental models of the human torso have been developed for ballistic impact testing. The human torso finite element model (HTFEM), including the thoracic skeletal structure and organs, was created in the finite element code LS-DYNA. The skeletal structure was assumed to be linear elastic while all internal organs were modeled as viscoelastic. A physical human surrogate torso model (HSTM) was developed using biosimulant materials and the same anthropometry as the HTFEM. The HSTM response to impact was recorded with piezoresistive pressure sensors molded into the heart, liver and stomach and an accelerometer attached to the sternum. For experimentation, the HSTM was outfitted with National Institute of Justice (NIJ) Level I, IIa, II and IIIa soft armor vests. Twenty-six ballistic tests targeting the HSTM heart and liver were conducted with 22 caliber ammunition at a velocity of 329 m/s and 9 mm ammunition at velocities of 332, 358 and 430 m/s. The HSTM pressure response repeatability was found to vary by less than 10% for similar impact conditions. A comparison of the HSTM and HTFEM response showed similar pressure profiles and less than 35% peak pressure difference for organs near the ballistic impact point. Furthermore, the peak sternum accelerations of the HSTM and HTFEM varied by less than 10% for impacts over the sternum. These models provide comparative tools for determining the thoracic response to ballistic impact and could be used to evaluate soft body armor design and efficacy, determine thoracic injury mechanisms and assist with injury prevention. PMID- 16376356 TI - Determination of trace levels of dinitrophenolic compounds in environmental water samples using hollow fiber supported liquid membrane extraction and high performance liquid chromatography. AB - A hollow fiber supported liquid membrane extraction method for the liquid chromatographic determination of dinitrophenolic compounds at ppt levels has been developed. Different variables affecting the extraction process, such as extraction time, shaking speed, acceptor pH, acceptor buffer concentration, salt content and humic acids have been studied. Enrichment factors up to 7000 times were obtained. Validation of the method included calibration experiments and studies of the linearity of the responses in different matrices. Good linearity was obtained in the environmental matrices evaluated. Detection limits range from 6.0 to 8.0 ng/L, and the relative standard deviations do not exceed 7% in terms of repeatability. PMID- 16376357 TI - Sedimentation velocity and potential in concentrated suspensions of charged porous spheres. AB - The body-force-driven migration in a homogeneous suspension of polyelectrolyte molecules or charged flocs in an electrolyte solution is analyzed. The model used for the particle is a porous sphere in which the density of the hydrodynamic frictional segments, and therefore also that of the fixed charges, is constant. The effects of particle interactions are taken into account by employing a unit cell model. The overlap of the electric double layers of adjacent particles is allowed and the relaxation effect in the double layer surrounding each particle is considered. The electrokinetic equations which govern the electrostatic potential profile, the ionic concentration (or electrochemical potential energy) distributions, and the fluid velocity field inside and outside the porous particle in a unit cell are linearized by assuming that the system is only slightly distorted from equilibrium. Using a regular perturbation method, these linearized equations are solved for a symmetrically charged electrolyte with the density of the fixed charges as the small perturbation parameter. An analytical expression for the settling velocity of the charged porous sphere is obtained from a balance among its gravitational, electrostatic, and hydrodynamic forces. A closed-form formula for the sedimentation potential in a suspension of identical charged porous spheres is also derived by using the requirement of zero net electric current. The dependence of the sedimentation velocity and potential of the suspension on the particle volume fraction and other properties of the particle-solution system is found to be quite complicated. PMID- 16376358 TI - Pressure dependence of the contact angle in a CO2-H2O-coal system. AB - Carbon dioxide injection into coal layers serves the dual purpose to enhance coal bed methane production (ECBM) and to store CO2. The efficiency of this process is expected to be much higher if water is the non-wetting phase in the coal-water gas system. Therefore, contact angles in the coal-water-CO2 system have been measured using the captive bubble technique in the pressure range between atmospheric pressure and 141 bar at a temperature of 45 degrees C. At atmospheric pressure the contact angle of a shrinking CO2 droplet increases with time, but stays below 90 degrees . At higher pressures (>2.6 bar) the contact angle increases beyond 90 degrees . The pressure dependence of the contact can be represented by theta=(111 degrees +/-10.5 degrees )+(0.17+/-0.14)P [bar]. The exceptional behavior at atmospheric pressure is possibly related to the stability of water patches on the coal surface. It is concluded that water is the non wetting phase in this coal-water-CO2 system. PMID- 16376359 TI - The temperature dependence of the heat capacity change for micellization of nonionic surfactants. AB - The thermodynamic parameters that govern micelle formation by four different nonionic surfactants were investigated by ITC and DSC. These included n dodecyldimethylphosphine oxide (APO12), Triton X-100 (TX-100), n octyltetraoxyethylene (C8E4), and N,N-dimethyloctylamine-N-oxide (DAO8). All of these surfactants had been previously investigated by solution calorimetry over smaller temperature ranges with conflicting conclusions as to the temperature dependence of the heat capacity change, DeltaCp, for the process. The temperature coefficient of the heat capacity change, B (cal/mol K2), was derived from the enthalpy data that were obtained at small intervals over a broad temperature range. The values obtained for each of the surfactants at 298.2 K for DeltaCp and B were -155+/-2 and 0.50+/-0.36 (APO12), -97+/-3 and -0.24+/-0.18 (TX-100), 105+/-2 and 1.0+/-0.3 (C8E4), and -82+/-1 and 0.36+/-0.04 (DAO8), cal/mol K and cal/mol K2, respectively. The resulting B-values did not correlate with the cmc, aggregation number, or structure of the monomer in an obvious way, but they were found to reflect the relative changes in hydration of the polar and nonpolar portions of the surfactant molecule as the micelles are formed. An analysis of the data obtained from DSC scans was used to describe the temperature dependence of the critical micelle concentration, cmc. An abrupt increase in heat capacity was observed for TX-100 and C8E4 solutions of 36.5+/-0.5 and 21+/-5 cal/mol K, respectively, as the temperature of the scan passed through the cloud point. This change in heat capacity may reflect the increased monomer concentration of the solutions that accompanies phase separation, although other interpretations of this jump are possible. PMID- 16376360 TI - Maximum stability of a single spherical particle attached to an emulsion drop. AB - The stability of a single, solid, spherical particle attached to a liquid drop in an emulsion is discussed. Ignoring effects due to gravity and line tension, we calculate the energies required to either detach the particle from the drop or to engulf the particle within the drop. The stability of the attached particle is here defined as the smaller of these two energies. A simple formula is derived for the value of Young's angle which gives maximum stability for a given radius ratio of particle to detached drop. For maximum stability the particle should be preferentially wetted by the liquid forming the drop. PMID- 16376361 TI - Eddies in a bottleneck: an arbitrary Debye length theory for capillary electroosmosis. AB - Using an applied electrical field to drive fluid flows becomes desirable as channels become smaller. Although most discussions of electroosmosis treat the case of thin Debye layers, here electroosmotic flow (EOF) through a constricted cylinder is presented for arbitrary Debye lengths (kappa(-1)) using a long wavelength perturbation of the cylinder radius. The analysis uses the approximation of small potentials. The varying diameter of the cylinder produces radially and axially varying effective electric fields, as well as an induced pressure gradient. We predict the existence of eddies for certain constricted geometries and propose the possibility of electrokinetic trapping in these regions. We also present a leading-order criterion which predicts central eddies in very narrow constrictions at the scale of the Debye length. Eddies can be found both in the center of the channel and along the perimeter, and the presence of the eddies is a consequence of the induced pressure gradient that accompanies electrically driven flow into a narrow constriction. PMID- 16376362 TI - Electroviscous cylinder-wall interactions. AB - A theoretical analysis is presented to determine the forces of interaction between an electrically charged cylindrical particle and a charged plane boundary wall when the particle translates parallel to the wall and rotates around its axis in a symmetric electrolyte solution at rest. The electroviscous effects, arising from the coupling between the electrical and hydrodynamic equations, are determined as a solution of three partial differential equations, derived from R.G. Cox's general theory [J. Fluid Mech. 338 (1997) 1], for electroviscous ion concentration, electroviscous potential, and electroviscous flow field. It is assumed a priori that the double layer thickness surrounding each charged surface is much smaller than the length scale of the problem. Using the matched asymptotic expansion technique, the electroviscous forces experienced by the cylinder are explicitly determined analytically for small particle-wall distances for low and intermediate Peclet numbers. It is found that the tangential force usually increases the drag above the purely hydrodynamic drag, although for certain conditions the drag can be reduced. Similarly the normal force is usually repulsive, i.e., it is an electrokinetic lift force, but under certain conditions the normal force can be attractive. PMID- 16376363 TI - Electrooptics of beta-FeOOH particle in aqueous media 2. Field-strength dependence of decay and steady-state birefringence, and the hydrodynamic and electrooptic properties. AB - Steady-state and decay birefringence, expressed in terms of the optical phase retardation per cell length delta/d, was measured on beta-FeOOH in aqueous ionic media at 633 nm and at 25 degrees C by an electric square-pulse technique over a wide range of field strength E to ca. 6 kV/cm. The field-strength dependence of both delta/d and field-free rotational relaxation time tau was determined at the sample concentrations between 0.0011 and 0.055 g/L and in the 0.02-2.0 mM NaCl concentration range. Extrapolation of both delta/d and tau values to infinitely high fields (E(2)-->infinity) could yield birefringence- and weight-average quantities, respectively. Observed tau values were decreased at weak fields but leveled off to ca. 0.3 ms at very high fields due to a slight polydispersity regarding the length and volume of particles. The weight-average relaxation time tau(w) was calculated with Perrin's expressions theoretically from the length, width, and volume of beta-FeOOH particles estimated in the dried state from electron micrograph. These quantities were variously averaged. The size distribution was discussed in terms of observed discrete histogram and theoretical (Weibull and Lansing-Kraemer) distribution functions. The sign of observed delta/d value was always positive. The infinitely high-field (delta/d)(infinity) values and the reduced optical anisotropy factor Delta g/n were evaluated by fitting to theoretical orientation functions. The intrinsic birefringence (n3-n1) could be estimated with the mean refractive index n(p) reported in the literature. For the spindle-shaped particle with an axial ratio of ca. 4, the sign of Delta g/n is always positive, whereas the quantity (n3-n1) was either negative (n(p) > 2.35) or positive (n(p) < 2.05) in sign or nearly zero (ca. n(p) = 2.26), depending critically on the n(p) values. PMID- 16376364 TI - Inner- and outer-sphere complexation of ions at the goethite-solution interface. AB - Formation of inner- and outer-sphere complexes of environmentally important divalent ions on the goethite surface was examined by applying the charge distribution CD model for inner- and outer-sphere complexation. The model assumes spatial charge distribution between the surface (0-plane) and the next electrostatic plane (1-plane) for innersphere complexation and between the 1 plane and the head end of the diffuse double layer (2-plane) for the outersphere complexation. The latter approach has been used because the distance of closest approach to a charged surface may differ for different ions. The surface structural approach implies the use of a Three-Plane model for the compact part (Stern layer) of solid-solution interface, which is divided into two layers. The thickness of each layer depends on the capacitance and the local dielectric constant. The new approach has been applied to describe the adsorption of magnesium, calcium, strontium, and sulfate ions. It is shown that the concept can successfully describe the development of surface charge in the presence of Ca(+2), Mg(+2), Sr(+2), and SO4(-2) as a function of loading, pH, and salt level, and also the shift in the isoelectric point (IEP) of goethite. The CD modeling revealed that, for the conditions studied, magnesium is mainly adsorbed as a bidentate innersphere complex, calcium can be a combination of bidentate innersphere and a monodentate inner- or outer-sphere complexes, and strontium is probably adsorbed as an outersphere complex. Sulfate is present as a mixture of inner- and outer-sphere monodentate complexes. Outersphere complexation is less pH dependent than innersphere complexation. The CD model predicts that the outersphere complexation of divalent cations and anions is relatively favorable at respectively low and high pH. Increase of ion loading favors the formation of innersphere complexes. PMID- 16376365 TI - Cloud point extraction and simultaneous determination of zirconium and hafnium using ICP-OES. AB - In the present study a simple versatile separation method using cloud point procedure for extraction of trace levels of zirconium and hafnium is proposed. The extraction of analytes from aqueous samples was performed in the presence of quinalizarine as chelating agent and Triton X-114 as a non-ionic surfactant. After phase separation, the surfactant-rich phase was diluted with 30% (v/v) propanol solution containing 1 mol l(-1) HNO3. Then, the enriched analytes in the surfactant-rich phase were determined by inductively coupled plasma-optical emission spectrometry (ICP-OES). The different variables affecting the complexation and extraction conditions were optimized. Under the optimum conditions (i.e. 3.4 x 10(-5) mol l(-1) quinalizarine, 0.1% (w/v) Triton X-114, 55 degrees C equilibrium temperature) the calibration graphs were linear in the range of 0.5-1000 mug l(-1) with detection limits (DLs) of 0.26 and 0.31 microg l(-1) for Zr and Hf, respectively. Under the presence of foreign ions no significant interference was observed. The precision (%RSD) for 8 replicate determinations at 200 microg l(-1) of Zr and Hf was better than 2.9% and the enrichment factors were obtained as 38.9 and 35.8 for Zr and Hf, respectively. Finally, the proposed method was successfully utilized for the determination of these cations in water and alloy samples. PMID- 16376366 TI - The generation of void morphology inside soap-free P(MMA-EA-MAA) particles prepared by seeded emulsion polymerization. AB - Monodisperse soap-free P(MMA-EA-MAA) latex particles were synthesized by seeded emulsion polymerization of methyl methacrylate (MMA), ethyl acrylate (EA) and methacrylic acid (MAA), and the particles with void morphology were obtained after undergoing alkali post-treatment. Effects of treatment conditions on particle morphology were investigated. Results showed that the void particles can be obtained under the conditions of the temperature >60 degrees C, initial pH >10.0, treatment time >20 min and 2-butanone amount >2.0 ml. The particle volume and the void size increased to the maximum and then decreased with the increases of initial pH and the treatment time, and these two values increased monotonously with the treatment temperature or 2-butanone amount increased. When the treatment temperature was elevated to 90 degrees C, the treatment time was longer than 180 min, or the 2-butanone amount was more than 8.0 ml, the relatively small voids inside most of the particles combined together to form a large one. The void structure disappeared completely as the initial pH was higher than 12.0. The generation mechanism of the void morphology was discussed. PMID- 16376367 TI - Microstructure and rheological properties of liquid crystallines formed in Brij 97/water/IPM system. AB - The phase diagram of Brij 97/water/IPM systems was determined at 25 degrees C. Rich liquid crystalline phases including Lalpha, H1, and cubic Fd3m phases were identified by means of small angle X-ray scattering (SAXS). Microstructure transitions of liquid crystals with changes in surfactant concentration and oil content are explained qualitatively by the surfactant packing parameter (vL/aSlc). Dynamic rheological results indicate that all three kinds of liquid crystals investigated show high elasticity. The lamellar, Lalpha, phases formed in Brij 97/water with two different oils, oleic acid and geraniol, were also studied in comparison with those of Brij 97/water/IPM systems. The strength of the network of lamellar phases formed in Brij 97/water/oleic acid and Brij 97/water/geraniol systems are appreciably stronger than for Brij 97/water/IPM systems, indicated by the smaller area of surfactant molecules at the interface and the higher moduli (G' and G''). PMID- 16376368 TI - Intelligibility of conversational speech produced by children with cochlear implants. AB - The intelligibility of conversational speech produced by six children fitted with cochlear implants before age 3 years was measured longitudinally. Samples were obtained every 3 months during periods of 12-21 months. Intelligibility was measured using both an utterance-by-utterance approach and an approach to the sample as a whole. Statistically significant differences were observed between the two approaches, but the differences were all within the realm of measurement error. Findings indicated that intelligible speech emerges quite rapidly in these children. Conversational intelligibility appears to be superior to that reported in the literature for similar children who use hearing aids but not necessarily as good as in children with normal hearing. Both intelligibility measures were significantly correlated with chronological age, hearing age, and amount of implant use, but were most strongly correlated with chronological age. LEARNING OUTCOMES: The reader will be able to (1) describe some of the issues involved in measuring speech intelligibility in children with cochlear implants and (2) describe the pattern of outcomes for the intelligibility of speech produced by children receiving cochlear implants before age 3 years. PMID- 16376369 TI - The effects of variation on learning word order rules by adults with and without language-based learning disabilities. AB - Non-adjacent dependencies characterize numerous features of English syntax, including certain verb tense structures and subject-verb agreement. This study utilized an artificial language paradigm to examine the contribution of item variability to the learning of these types of dependencies. Adult subjects with and without language-based learning disabilities listened to strings of three non words for which the first and third elements had a dependent relationship. In the low variability condition, 12 non-words occurred in the middle position, and in the high variability condition, 24 non-words occurred in this position. Non disabled adults were able to learn the non-adjacent contingencies and generalize the underlying structure to new strings, but only when variability was high. Adults with language-based learning disabilities did not perform above chance levels under either variability condition. Thus, this group showed poor sensitivity to statistical information in speech input that both infants and non disabled adults are known to track. LEARNING OUTCOMES: As a result of this activity, the reader will: (1) understand the advantages of using an artificial language to investigate language learning; (2) become familiar with a paradigm for studying the rapid learning of syntactic contingencies; (3) comprehend how the ability to map language structure differs for non-disabled adults and adults with a history of language/learning disability as a function of variability in the input the listener receives. PMID- 16376370 TI - The representation of multiplication facts: developmental changes in the problem size, five, and tie effects. AB - In this study, we investigated the development of basic effects that have been found in single-digit multiplication arithmetic: the problem size, five, and tie effects. Participants (9-, 10-, and 11-year-olds and adults) performed a production task on simple multiplication. The procedure replicated study [Canadian Journal of Psychology, Vol. 39, pp. 338-366], but the results show that the gradual decrease of the problem size effect ends in sixth grade. We report analyses on raw latencies and state trace analyses that take into account reaction time scaling as a function of age. The results show that 11-year-olds do not differ significantly from adults on any of the three effects. Before 11 years of age, interesting developmental changes occur. PMID- 16376371 TI - An enzyme-linked immunosorbent assay for human cathepsin X, a potential new inflammatory marker. AB - The human lysosomal cysteine-type carboxypeptidase cathepsin X is mainly present in monocytes and macrophages and may be released into the circulation due to constitutive and/or regulated secretion by (activated) immune cells. To define its potential diagnostic value as an inflammatory marker, we have developed a highly sensitive and specific sandwich-type immunoassay (ELISA) for cathepsin X permitting both intra- and extracellular detection and quantification. The dynamic range of the cathepsin X ELISA was determined to be 100 (detection limit) to 8000 pg/ml. Reproducibility of both within and between runs yielded coefficients of variation (CVs) of 2.7-3.5% and 6.3-7.3%, respectively. Cross reactivity with other members (cathepsin B, L) of the thiol-dependent cathepsin family was not observed. The ELISA was used to quantify cathepsin X in leukocytes as well as in plasma of healthy volunteers and patients with multiple trauma. During the first 72 h after trauma, plasma levels of cathepsin X increased significantly, particularly in patients who died during the posttraumatic period. In comparison to the well-known inflammation marker neutrophil elastase, cathepsin X levels predicted survival with a higher significance in the later posttraumatic phase. In conclusion, this report provides the first evidence of cathepsin X immunoreactivity not only in cell lysates but also in plasma samples. We suggest that the newly developed highly reproducible ELISA will be of great value for further evaluation of this protease as a diagnostic and/or prognostic marker in inflammatory diseases. PMID- 16376372 TI - Mapping of FVIII inhibitor epitopes using cellulose-bound synthetic peptide arrays. AB - Epitope mapping using antibodies against factor VIII (FVIII) has been performed using blotting techniques with truncated and/or digested FVIII molecules. Here, we focused on the precise mapping of affinity purified IgG from patients with an immune response against blood clotting FVIII using synthetic peptide arrays on cellulose membranes comprising the entire sequence of FVIII. The aim was to elucidate the epitope profile from different inhibitors and possibly detect new epitopes, which have not been described before. The epitope patterns from five patients showed reactivity with all domains in the FVIII molecule, but were different between various patients. These results included epitopes usually buried within the folded protein. However, in competition assays using FVIII as competitive agent in a mixture with inhibitor IgG, the most immunogenic regions were located in the FVIII light chain. Our results show that the C1 domain was the region with highest immunogenicity in all patients. Here, we demonstrate that the SPOT method is very well suited for the precise location of epitopes in the core of the protein, which usually cannot be detected by other methods. PMID- 16376373 TI - Specific and sensitive detection of Nosema bombi (Microsporidia: Nosematidae) in bumble bees (Bombus spp.; Hymenoptera: Apidae) by PCR of partial rRNA gene sequences. AB - A polymerase chain reaction (PCR) based method was developed for the specific and sensitive diagnosis of the microsporidian parasite Nosema bombi in bumble bees (Bombus spp.). Four primer pairs, amplifying ribosomal RNA (rRNA) gene fragments, were tested on N. bombi and the related microsporidia Nosema apis and Nosema ceranae, both of which infect honey bees. Only primer pair Nbombi-SSU-Jf1/Jr1 could distinguish N. bombi (323bp amplicon) from these other bee parasites. Primer pairs Nbombi-SSU-Jf1/Jr1 and ITS-f2/r2 were then tested for their sensitivity with N. bombi spore concentrations from 10(7) down to 10 spores diluted in 100 microl of either (i) water or (ii) host bumble bee homogenate to simulate natural N. bombi infection (equivalent to the DNA from 10(6) spores down to 1 spore per PCR). Though the N. bombi-specific primer pair Nbombi-SSU-Jf1/Jr1 was relatively insensitive, as few as 10 spores per extract (equivalent to 1 spore per PCR) were detectable using the N. bombi-non-specific primer pair ITS f2/r2, which amplifies a short fragment of approximately 120 bp. Testing 99 bumble bees for N. bombi infection by light microscopy versus PCR diagnosis with the highly sensitive primer pair ITS-f2/r2 showed the latter to be more accurate. PCR diagnosis of N. bombi using a combination of two primer pairs (Nbombi-SSU Jf1/Jr1 and ITS-f2/r2) provides increased specificity, sensitivity, and detection of all developmental stages compared with light microscopy. PMID- 16376374 TI - New cell lines from larval fat bodies of Spodoptera exigua: characterization and susceptibility to baculoviruses (Lepidoptera: Noctuidae). AB - Two new cell lines, designated IOZCAS-Spex-II and IOZCAS-Spex-III, were initiated from the fat bodies of larvae of the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae) in TNM-FH medium containing 10% fetal bovine serum. The spherical cells were predominant among the various cell types and measures approximately 15 microm in diameter. The cell lines were mainly composed of tetraploid cells with chromosome numbers ranging from 116 to 131 (n=31). The cell lines were confirmed to have originated from the S. exigua by DAF-PCR technique. They were susceptible to the multiple nucleocapsid nuclear polyhedrosis viruses from S. exigua. PMID- 16376375 TI - Infection of the malaria mosquito Anopheles gambiae with the entomopathogenic fungus Metarhizium anisopliae reduces blood feeding and fecundity. AB - The entomopathogenic fungus Metarhizium anisopliae is being considered as a biocontrol agent against adult African malaria vectors. In addition to causing significant mortality, this pathogen is known to cause reductions in feeding and fecundity in a range of insects. In the present study we investigated whether infection with M. anisopliae affected blood feeding and fecundity of adult female malaria vectors Anopheles gambiae Giles sensu stricto. Mosquitoes were contaminated with either a low or a moderately high dose of oil-formulated conidia of M. anisopliae, and offered a single human blood meal 48, 72, or 96 h later to assess feeding propensity and individual blood meal size. In a second experiment, individual fungus-infected females were offered a blood meal every third day (to a total of 8 gonotrophic cycles), and allowed to oviposit after each cycle in order to quantify feeding propensity and fecundity. Infected females took smaller blood meals and displayed reduced feeding propensity. It was found that mosquitoes, inoculated with a moderately high dose of fungal conidia, exhibited reduced appetite related to increasing fungal growth. Of the fungus infected females, the proportion of mosquitoes taking the second blood meal was reduced with 51%. This was further reduced to 35.3% by the 4th blood meal. During 8 feeding opportunities, the average number of blood meals taken by uninfected females was 4.39, against 3.40 (low dose), and 2.07 (high dose) blood meals for the fungus-infected females. Moreover, infected females produced fewer eggs per gonotrophic cycle and had a lower life-time fecundity. Epidemiological models show that both blood feeding and fecundity are among the most important factors affecting the likelihood of a mosquito transmitting malaria, which suggests that this fungus may have potential as biocontrol agent for vector-borne disease control. PMID- 16376376 TI - Solvation-assisted pressure tuning of insulin fibrillation: from novel aggregation pathways to biotechnological applications. AB - Solvation-assisted pressure tuning has been employed to unravel unknown structural and kinetic aspects of the insulin aggregation and fibrillation process. Our approach, using fluorescence, Fourier transform infrared and atomic force microscopy techniques in combination with pressure and solvent perturbation, reveals new insights into the pre-aggregated regime as well as mechanistic details about two concurrent aggregation pathways and the differential stability of insulin aggregates. Pressure uniformly fosters the dissociation of native insulin oligomers, whereas the aggregation pathways at elevated temperatures are affected by pressure differently and in a cosolvent dependent manner. Moderate pressures accelerate the amyloid pathway in the presence of EtOH (leading to essentially monomeric aggregating species) via relatively dehydrated transition states with negative activation volumes for nucleation and elongation. Alternatively, a novel, fast equilibrium pathway to distinct beta-sheet-rich oligomers with thioflavin T-binding capability is accessible to partially unfolded insulin monomers at pressures below approximately 200 bar in the absence of EtOH. These oligomers, probably off the normal fibrillation pathway, are stabilized mainly by electrostatic and hydrophobic interactions, lacking the precise packing of mature insulin fibrils, which renders them susceptible to quantitative pressure-induced dissociation. Due to a highly negative activation volume for dissociation (-70(+/-16)ml/mol), pressure dissociation is fast and technologically feasible at ambient temperatures and moderate pressures. Becoming kinetically very labile above 35 degrees C, the pressurized oligomers can re-enter the slower, ultimately irreversible, fibrillation pathway at higher temperatures. At pressures above approximately 1000 bar, the partial unfolding of insulin monomers, accompanied by a volumetric expansion, dominates the aggregation kinetics, which manifests in a progressive inhibition of the fibrillation. Unlike their precursors, the pressure insensitivity of mature insulin fibrils demonstrates that an extensive hydrogen bonding network and optimized side-chain packing are crucial for their stability. PMID- 16376377 TI - Influence of the 2'-hydroxyl group conformation on the stability of A-form helices in RNA. AB - The 2'-hydroxyl group plays fundamental roles in both the structure and the function of RNA, and is the major determinant of the conformational and thermodynamic differences between RNA and DNA. Here, we report a conformational analysis of 2'-OH groups of the HIV-2 TAR RNA by means of NMR scalar coupling measurements in solution. Our analysis supports the existence of a network of water molecules spanning the minor groove of an RNA A-form helix, as has been suggested on the basis of a high-resolution X-ray study of an RNA duplex. The 2' OH protons of the lower stem nucleotides of the TAR RNA project either towards the O3' or towards the base, where the 2'-OH group can favorably participate in H bonding interactions with a water molecule situated in the nucleotide base plane. We observe that the k(ex) rate of the 2'-OH proton with the bulk solvent anti correlates with the base-pair stability, confirming the involvement of the 2'-OH group in a collective network of H-bonds, which requires the presence of canonical helical secondary structure. The methodology and conformational analysis presented here are broadly applicable and facilitate future studies aimed to correlate the conformation of the 2'-OH group with both the structure and the function of RNA and RNA-ligand complexes. PMID- 16376378 TI - GTP-dependent formation of a ribonucleoprotein subcomplex required for ribosome biogenesis. AB - Ribosome biogenesis in eukaryotic organisms involves the coordinated assembly of 78 ribosomal proteins onto the four ribosomal RNAs, mediated by a host of trans acting factors whose specific functions remain largely unknown. The essential GTPase Bms1, the putative endonuclease Rcl1 and the essential U3 small nucleolar RNA form a stable subcomplex thought to control an early step in the assembly of the 40S ribosomal subunit. Here, we provide a complete thermodynamic analysis of GTP-dependent subcomplex formation, revealing strong thermodynamic coupling of Rcl1, U3 small nucleolar RNA and GTP binding to Bms1 that is eliminated in the presence of GDP. The results suggest that Rcl1 activates Bms1 by promoting GDP/GTP exchange, analogous to ribosome-promoted nucleotide exchange within translation elongation factor EF-G. These and other data unveil thermodynamic similarities between Bms1 and the subgroup of GTPases involved in translation, providing evidence that parts of the ribosome assembly machinery may have evolved from the translation apparatus. This quantitative description of an early and essential step in pre-ribosome assembly provides a framework for elucidating the network of interactions between the Bms1 subcomplex and additional factors involved in ribosome biogenesis. PMID- 16376379 TI - DNA recombination activity in soybean mitochondria. AB - Mitochondrial genomes in higher plants are much larger and more complex as compared to animal mitochondrial genomes. There is growing evidence that plant mitochondrial genomes exist predominantly as a collection of linear and highly branched DNA molecules and replicate by a recombination-dependent mechanism. However, biochemical evidence of mitochondrial DNA (mtDNA) recombination activity in plants has previously been lacking. We provide the first report of strand invasion activity in plant mitochondria. Similar to bacterial RecA, this activity from soybean is dependent on the presence of ATP and Mg(2+). Western blot analysis using an antibody against the Arabidopsis mitochondrial RecA protein shows cross-reaction with a soybean protein of about 44 kDa, indicating conservation of this protein in at least these two plant species. mtDNA structure was analyzed by electron microscopy of total soybean mtDNA and molecules recovered after field-inversion gel electrophoresis (FIGE). While most molecules were found to be linear, some molecules contained highly branched DNA structures and a small but reproducible proportion consisted of circular molecules (many with tails) similar to recombination intermediates. The presence of recombination intermediates in plant mitochondria preparations is further supported by analysis of mtDNA molecules by 2-D agarose gel electrophoresis, which indicated the presence of complex recombination structures along with a considerable amount of single-stranded DNA. These data collectively provide convincing evidence for the occurrence of homologous DNA recombination in plant mitochondria. PMID- 16376382 TI - Position preference on glucuronidation of mono-hydroxylflavones in human intestine. AB - Extensive intestinal glucuronidation has been previously reported in both human and animals after oral administration of naturally occurred flavonoids. The present study aims to investigate the relationship between human intestinal glucuronidation activity and the position of hydroxyl substitution on flavonoids. Seven commercially available mono-hydroxyflavones (HF), namely 3-, 5-, 6-, 7-, 2' , 3'- and 4'-mono-hydroxyflavones, were chosen as model compounds. Glucuronidation activity of the selected seven HFs was investigated by incubating each HF at various concentrations with human jejunum S9 at 37 degrees C for 10 min. The generated glucuronides were identified by HPLC/MS and quantified by HPLC/UV. Metabolic kinetics parameters including Km and Vmax of each HF were determined. The results demonstrated that the glucuronidation activity of 6- and 3'-mono-hydroxyflavones was much greater than that of 3-, 4'-, 7- and 2'-HF with 5-HF to be the lowest. The findings imply that nucleophilicity and stereo conformation of OH substituents are crucial for the intestinal glucuronidation of flavonoids. PMID- 16376383 TI - Characteristics and inhibition by flavonoids of 20alpha-hydroxysteroid dehydrogenase activity in mouse tissues. AB - Progesterone was stereoselectively reduced to a metabolite 20alpha-hydroxy-4 pregnen-3-one in the cytosolic fraction from the liver of male mice, indicating that the reduction of progesterone is catalyzed by 20alpha-hydroxysteroid dehydrogenase (20alpha-HSD). The cytosolic 20alpha-HSD activity was observed not only in the liver, but also in the kidney and lung. In liver cytosol, both NADPH and NADH were effective as cofactors for 20alpha-HSD activity, although NADPH was better than NADH for the enzyme activity. On the other hand, 20alpha-HSD activity in kidney cytosol required only NADPH as a cofactor. No significant sex-related difference of 20alpha-HSD activity was observed in liver and kidney cytosols. Flavonoids have been reported to inhibit the biosynthesis and metabolism of steroids. However, little is known about inhibitory effects of flavonoids on 20alpha-HSD activity. Thus, the effects of 16 flavonoids on 20alpha-HSD activity were examined, using liver cytosol of male mice. Among flavonoids tested, fisetin, apigenin, naringenin, luteolin, quercetin and kaempferol exhibited high inhibitory potencies for the 20alpha-HSD activity. We propose the possibility that these flavonoids augment progesterone signaling by inhibiting potently 20alpha-HSD activity in non-reproductive tissues. PMID- 16376384 TI - Expression of glucocorticoid receptor in the intestine of a euryhaline teleost, the Mozambique tilapia (Oreochromis mossambicus): effect of seawater exposure and cortisol treatment. AB - Cortisol plays an important role in controlling intestinal water and ion transport in teleosts possibly through glucocorticoid receptor (GR) and/or mineralocorticoid receptor. To better understand the role of GR in the teleost intestine, in a euryhaline tilapia, Oreochromis mossambicus, we examined (1) the intestinal localizations of GR; (2) the effects of environmental salinity challenge and cortisol treatment on GR mRNA expression. The mRNA abundance of GR in the posterior intestinal region of tilapia was found to be higher than that in the anterior and middle intestine. In the posterior intestine, GR appears to be localized in the mucosal layer. GR mRNA levels in the posterior intestine were elevated after exposure of freshwater fish to seawater for 7 days following an increase in plasma cortisol. Similarly, cortisol implantation in freshwater tilapia for 7 days elevated the intestinal GR mRNA. These results indicate that seawater acclimation is accompanied by upregulation of GR mRNA abundance in intestinal tissue, possibly as a consequence of the elevation of cortisol levels. In contrast, a single intraperitoneal injection of cortisol into freshwater tilapia decreased intestinal GR mRNA. This downregulation of the GR mRNA by cortisol suggests a dual mode of autoregulation of GR expression by cortisol. PMID- 16376385 TI - Cardiovascular, anthropometric and neurocognitive features of healthy postmenopausal women: effects of hormone replacement therapy. AB - Randomized clinical trials have not shown long-term benefit of postmenopausal hormone replacement therapy (PHT) nor have they shown conclusively that the harmful consequences outweighs the benefits of the treatment. Rather, it is possible that an individualized hormone replacement therapy in questionably clinically healthy postmenopausal women may lead to different results than randomized trials. DESIGN: In this cross-sectional study we evaluated anthropometric parameters, body composition, serum lipids, blood pressure, heart rate variability (HRV) and neurocognitive functions in 39 healthy postmenopausal women PHT users or not users (n=13, age 53.0+/-3.3 and n=26, age=53.3+/-5.0 SD, respectively) as well as in 27 younger controls (ages=33.3+/-7.1). RESULTS: Demographic parameters were similar in women PHT users and not users. Postmenopausal women showed a significantly increase of body mass index (BMI) as well as of waist circumference, compared to younger controls, but in PHT users the values of fat free mass were intermediate between the ones of not treated and younger women. The study of HRV showed a reduction in low frequency (LF) component (sympathetic modulation) during the day, and a reduction in high frequency (HF) component (parasympathetic modulation), particularly in postmenopausal women without PHT. PHT users were characterized by autonomic parameters intermediate between younger controls and age-matched women without PHT. CONCLUSIONS: The impact of PHT on the age-dependent changes of anthropometric features and body composition seems to be modest but positive. Furthermore, PHT seems to play a positive role on the autonomic modulation of cardiac function, through a shift of LF/HF ratio values towards those of young controls. PMID- 16376386 TI - Scoparone inhibits PMA-induced IL-8 and MCP-1 production through suppression of NF-kappaB activation in U937 cells. AB - Scoparone is a major component of the shoot of Artemisia capillaris (Compositae), which has been used for the treatment of hepatitis and biliary tract infection in oriental countries. In this study, the effects of scoparone on the expression of interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1) and activation of nuclear factor-kappaB (NF-kappaB) were examined in U937 human monocytes activated with phorbol 12-myristate 13-acetate (PMA). Scoparone (5-100 microM) had no cytotoxic effect in unstimulated cells and concentration-dependently reversed PMA induced toxicity in the cells stimulated with PMA. Scoparone concentration dependently reduced the release of IL-8 and MCP-1 protein and expression of IL-8 and MCP-1 mRNA levels induced by PMA. Moreover, scoparone inhibited the levels of NF-kappaB-DNA complex and NF-kappaB activity in the cells stimulated with PMA in a concentration-dependent manner. Scoparone dose-dependently inhibited the phosphorylation of IkappaBalpha and nuclear translocation of NF-kappaB1 p50, RelA p65, and c-Rel p75. These data suggest that scoparone may inhibit the expression of chemokines (IL-8 and MCP-1) in PMA-stimulated U937 cells and a potential mechanism of scoparone may be inhibition of NF-kappaB activation, which is linked to inhibition of NF-kappaB subunits (NF-kappaB1 p50, RelA p65, and c-Rel p75) translocation via suppression of IkappaBalpha phosphorylation. PMID- 16376387 TI - A survey of organic solvent extractable metal concentrations in the bottom sediments in Osaka Bay, Japan. PMID- 16376388 TI - Characterization of cycA mutants of Escherichia coli. An assay for measuring in vivo mutation rates. AB - Quantitative assessment of the spontaneous or induced genomic mutation rate, a fundamental evolutionary parameter, usually requires the use of well characterized mutant selection systems. Although there is a great number of genetic selection schemes available in Escherichia coli, the selection of D cycloserine resistant mutants is shown here to be particularly useful to yield a general view of mutation rates and spectra. The combination of a well-defined experimental protocol with the Ma-Sandri-Sarkar maximum likelihood method of fluctuation analysis results in reproducible data, adequate for statistical comparisons. The straightforward procedure is based on a simple phenotype genotype relationship, and detects mutations in the single-copy, chromosomal cycA gene, involved in the uptake of D-cycloserine. In contrast to the widely used rifampicin resistance assay, the procedure selects mutations which are neutral in respect of cell growth. No specific genetic background is needed, and practically the entire mutation spectrum (base substitutions, frameshifts, deletions, insertions) can simultaneously be measured. A systematic analysis of cycA mutations revealed a spontaneous mutation rate of 6.54 x 10(-8) in E. coli K-12 MG1655. The mutation spectrum was dominated by point mutations (base substitutions, frameshifts), spread over the entire gene. IS insertions, caused by IS1, IS2, IS3, IS4, IS5 and IS150, represented 24% of the mutations. PMID- 16376389 TI - Effects of selenium depletion and selenium repletion by choice feeding on selenium status of young and old laying hens. AB - We investigated the effect of choice feeding two diets with different selenium (Se) content to young and old moderately Se-deficient laying hens on serum Se (SSe), glutathione peroxidase (GPX), vitamin E, creatine kinase (CK), aspartate aminotransferase (ASAT), thyroxine (T4) and triiodothyronine (T3). Each of two consecutive study parts (I and II) with the same hens and treatments began with a 6-week baseline period (Medium-Se diet), followed by a 9-week depletion period (Low-Se or Medium-Se diet), followed by a 6-week choice period with two different diets offered simultaneously (Medium-Se/Low-Se, Medium-Se/High-Se, or Low-Se/High Se). During both depletion periods, SSe and GPX gradually decreased, whereas T4 gradually increased in hens fed Low-Se confirming gradual Se-depletion. T3 decreased transiently in young hens only. As reported earlier, Se-deficient hens preferred High-Se over Low-Se diet during the first 3 weeks of choice feeding in part I, not however in part II. This preference resulted in higher SSe in these hens. GPX activity did not reflect feed preference, probably because Se-intake exceeded Se-requirement for maximal GPX activity. In Part II, hens depleted with Low-Se diet had higher SSe when previously offered High-Se diet in either combination, than when offered Low-Se/Medium-Se, presumably due to Se-stores built during choice feeding in part I, which possibly prevented development of Se deficiency in part II. In addition, in older hens, Se depletion proceeded faster, whereas Se-repletion by choice feeding was slower than in young hens, indicating the increase in Se requirement with advancing age. Vitamin E, ASAT and CK remained largely unchanged by the treatments. PMID- 16376390 TI - Gastric compliance in bulimia nervosa. AB - Bulimia nervosa (BN) is a psychiatric illness characterized by eating binges followed by inappropriate behavioral attempts to compensate for the binges, usually vomiting or laxative abuse. Patients with BN have disturbances in the development of satiety during a meal as well as disturbances in functions of the upper gastrointestinal tract such as slowed gastric emptying, impaired gastric accommodation reflex and blunted cholecystokinin release. The present study examined gastric compliance and sensory responses to gastric distention in women with BN and controls. Sixteen women with BN and 13 healthy control subjects swallowed an inflatable bag that was placed in the proximal stomach. The bag was inflated to produce increasing steps of pressure against the stomach wall, before and after consumption of a 200 ml (200 Kcal) liquid meal. Pressure and volume were recorded for 2-min periods, beginning at 0 mm Hg pressure and increasing in steps of 2 mm Hg until subjects reported discomfort, gastric volume reached 600 ml, or pressure reached 20 mm Hg. At each pressure step subjects made sensory ratings. Gastric compliance was calculated as the slope of the best-fit straight line of each subject's gastric volume vs. gastric pressure. There was a significant postmeal increase in gastric compliance in both groups of subjects but there was no difference in compliance between patients with BN and controls. Patients with BN appeared to have diminished sensitivity to gastric distention. In conclusion, although other studies have described gastrointestinal abnormalities associated with BN, the current study found gastric compliance of patients with BN to be normal. PMID- 16376391 TI - Stilbenoids of Kobresia nepalensis (Cyperaceae) exhibiting DNA topoisomerase II inhibition. AB - Resveratrol oligomers, nepalensinol A, B and C, were isolated from the stem of Kobresia nepalensis (Cyperaceae). The structures were established on the basis of chemical properties and spectroscopic evidence including 2D NMR spectroscopic analysis. Nepalensinol A, B and C showed a potent inhibitory effect on topoisomerase II -- stronger than etoposide (VP-16), a topoisomerase II inhibitor used as an anti-cancer drug. Nepalensinol B, in particular, exhibited the most potent activity with an IC(50) of 0.02 microg/ml. PMID- 16376392 TI - Polyphenoloxidase activity in coffee leaves and its role in resistance against the coffee leaf miner and coffee leaf rust. AB - In plants, PPO has been related to defense mechanism against pathogens and insects and this role was investigated in coffee trees regarding resistance against a leaf miner and coffee leaf rust disease. PPO activity was evaluated in different genotypes and in relation to methyl-jasmonate (Meja) treatment and mechanical damage. Evaluations were also performed using compatible and incompatible interactions of coffee with the fungus Hemileia vastatrix (causal agent of the leaf orange rust disease) and the insect Leucoptera coffeella (coffee leaf miner). The constitutive level of PPO activity observed for the 15 genotypes ranged from 3.8 to 88 units of activity/mg protein. However, no direct relationship was found with resistance of coffee to the fungus or insect. Chlorogenic acid (5-caffeoylquinic acid), the best substrate for coffee leaf PPO, was not related to resistance, suggesting that oxidation of other phenolics by PPO might play a role, as indicated by HPLC profiles. Mechanical damage, Meja treatment, H. vastatrix fungus inoculation and L. coffeella infestation caused different responses in PPO activity. These results suggest that coffee resistance may be related to the oxidative potential of the tissue regarding the phenolic composition rather than simply to a higher PPO activity. PMID- 16376393 TI - Saponins from the seeds of Mimusops laurifolia. AB - Nine saponins were isolated from the seeds of Mimusops laurifolia. Their structures were established using one- and two-dimensional NMR spectroscopy and mass spectrometry. Three of them are identified as: 3-O-(beta-d-apiofuranosyl-(1- >3)-beta-d-glucuronopyranosyl)-28-O-(alpha-l-rhamnopyranosyl-(1-->3)-beta-d xylopyranosyl-(1-->4)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranosyl) 16alpha-hydroxyprotobassic acid, 3-O-(beta-d-glucopyranosyl-(1-->3)-beta-d glucopyranosyl)-28-O-(alpha-l-rhamnopyranosyl-(1-->3)-beta-d-xylopyranosyl-(1- >4)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranosyl)-16alpha hydroxyprotobassic acid and 3-O-(beta-d-glucopyranosyl-(1-->6)-beta-d glucopyranosyl-(1-->6)-beta-d-glucopyranosyl)-28-O-(alpha-l-rhamnopyranosyl-(1- >3)-beta-d-xylopyranosyl-(1-->4)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l arabinopyranosyl)-16alpha-hydroxyprotobassic acid. PMID- 16376394 TI - Flavonols and an indole alkaloid skeleton bearing identical acylated glycosidic groups from yellow petals of Papaver nudicaule. AB - From yellow petals of Iceland poppy, besides the known flavonoid gossypitrin, seven kaempferol derivatives were isolated. In addition to kaempferol 3-O-beta sophoroside and kaempferol 3-O-beta-sophoroside-7-O-beta-glucoside, known from other plants, the mono- and dimalonyl conjugates of the latter were identified by MS and NMR spectroscopy. Structure analyses of a set of co-occurring pigments, the nudicaulins, revealed that they have the identical acylated glycoside moieties attached to a pentacyclic indole alkaloid skeleton for which the structure of 19-(4-hydroxyphenyl)-10H-1,10-ethenochromeno[2,3-b]indole-6,8,18 triol was deduced from MS and NMR as well as chemical and chiroptical methods. PMID- 16376395 TI - Comparison of ventral coccygeal arterial and jugular venous blood samples for pH, pCO2, HCO3, Be(ecf) and ctCO2 values in calves with pulmonary diseases. AB - The aim of this study was to determine whether venous blood samples can be used as an alternative to arterial samples in calves with respiratory problems and healthy calves. Jugular vein and ventral coccygeal artery were used to compare blood gas values. Sampling of the jugular vein followed soon after sampling of the ventral coccygeal artery in healthy calves (group I) and calves with respiratory problems (group II). Mean values of arterial blood for pH, pCO2, HCO3act in healthy calves were 7.475 +/- 0.004, 4.84 +/- 0.2 kPa, 28.45 +/- 1.30 mmol/L compared with venous samples, 7.442 +/- 0.006, 6 +/- 0.3 kPa, 30.93 +/- 1.36 mmol/L, respectively. In group II, these parameters were 7.414 +/- 0.01, 5.93 +/- 0.3, 27.73 +/- 1.96 mmol/L for arterial blood and 7.398 +/- 0.008, 6.85 +/- 0.2 kPa, 29.77 +/- 1.91 mmol/L for venous blood, respectively. There were no statistically significant differences between arterial and venous pH, HCO3act, Be(ecf), ctCO2 values with the exception of pCO2 (P = 0.001) in group II. In group I, correlation (r2) between arterial and venous blood pH, pCO2, HCO3act were 84.5%, 87.5%, 95.7%, respectively compared with the same parameters in group II, 80.8%, 77.1%, 70.3%. In conclusion, venous blood gas values can predict arterial blood gas values of pH, pCO2 and HCO3ecf, Be(ecf) and ctCO2- for healthy calves but only pH values in calves with acute respiratory problems (r2 value>80%). PMID- 16376396 TI - Helicobacter pylori-associated chronic atrophic gastritis involving the gastric body and severe disease by Vibrio cholerae. AB - Evidence has associated chronic infection by Helicobacter pylori with chronic gastritis, low gastric acid production and an increased risk of life-threatening cholera. However, the relationship of specific patterns of histological damage in the gastric mucosa associated with H. pylori infection and the occurrence of cholera has not been described. The purpose of this study was to compare the gastric pH and histopathological findings in gastric biopsies taken from patients with severe diarrhoeal disease due to Vibrio cholerae with those taken from a control (cholera-negative) population. Thirty-five H. pylori-positive patients who had severe dehydration from culture-confirmed cholera (cases) and 40 patients with H. pylori but with no history of cholera (controls) were recruited. Gastric pH was measured and multiple biopsies were taken from the gastric antrum and body for histopathological examination. The results revealed that patients with severe cholera had a significantly higher prevalence of hypochlorhydria at endoscopy compared with controls. Furthermore, cases had significantly more chronic atrophic gastritis (45.7% vs. 12.5%; P=0.002) and intestinal metaplasia (37.1% vs. 2.5%; P<0.01) in the gastric body than controls. Our findings suggest that the nature and location of these gastric lesions may predispose a subset of H. pylori-infected individuals to severe disease by V. cholerae. PMID- 16376397 TI - Hormesis may provide a central concept for homeopathy development. PMID- 16376398 TI - Measurements of velocity and attenuation of leaky waves using an ultrasonic array. AB - A new method of measuring velocity and attenuation of leaky surface waves is presented. A single focused transmitting transducer and linear receiving array in a pitch-catch arrangement are used in the proposed system. The spatial distribution of the acoustic field in the leaky wave is recorded by the array, and the parameters of the leaky wave can be obtained by processing the output waveforms. In comparison with existing material characterization systems, the mechanical scanning of the transducers is not used any more, and the measurement time is only limited by the time of the wave propagation and speed of the electronic data acquisition system. PMID- 16376399 TI - Effect of adhesion force between crack planes on subharmonic and DC responses in nonlinear ultrasound. AB - Subharmonic and DC responses in nonlinear ultrasound have been expected as a possible means of detecting closed cracks. Recently, it has been reported that subharmonics in a closed crack markedly increases above a certain input wave amplitude. Such a phenomenon is called "threshold behavior". However, the mechanism of threshold behavior has yet to be elucidated. To clarify this, we introduced adhesion force as a short-range force into the previous analytical model, which expresses the nonlinear contact vibrations of crack planes with intense ultrasound and provides a DC displacement as an approximation of the subharmonic response. Consequently, upward convex curves of displacement against input wave amplitude above the threshold were reproduced for the first time. The validity of the derived analytical solution is discussed by comparison with experimentally observed subharmonics. PMID- 16376400 TI - Acoustic streaming in lithotripsy fields: preliminary observation using a particle image velocimetry method. AB - This study considers the acoustic streaming in water produced by a lithotripsy pulse. Particle image velocimetry (PIV) method was employed to visualize the acoustic streaming produced by an electromagnetic shock wave generator using video images of the light scattering particles suspended in water. Visualized streaming features including several local peaks and vortexes around or at the beam focus were easily seen with naked eyes over all settings of the lithotripter from 10 to 18 kV. Magnitudes of the peak streaming velocity measured vary in the range of 10-40 mm s(-1) with charging voltage settings. Since the streaming velocity was estimated on the basis of a series of the video images of particles averaged over 1/60s, the time resolution limited by the video frame rate which is 1-2 orders of magnitude larger than driving acoustic activities, measured velocities are expected to be underestimated and were shown a similar order of magnitude lower than those calculated from a simple theoretical consideration. Despite such an underestimation, it was shown that, as predicted by theory, the magnitude of the streaming velocity measured by the present PIV method was proportional to acoustic intensity. In particular it has almost a linear correlation with peak negative pressures (r=0.98683, p=0.0018). PMID- 16376401 TI - Developmental asymmetries between horizontal and vertical tracking. AB - The development of the asymmetry between horizontal and vertical eye tracking was investigated longitudinally at 5, 7, and 9 months of age. The target moved either on a 2D circular trajectory or on a vertical or horizontal 1D sinusoidal trajectory. Saccades, smooth pursuit, and head movements were measured. Vertical tracking was found to be inferior to horizontal tracking at all age levels. The results also show that the mechanisms responsible for horizontal and vertical tracking mutually influence one another in the production of 2D visual pursuit. Learning effects were observed within-trials but no transfer between trials was found. PMID- 16376402 TI - Mirror-image symmetry and search asymmetry: a comparison of their effects on visual search and a possible unifying explanation. AB - Visual search may be affected by mirror-image symmetry between target and non targets and also by switching the roles of target and non-target. Do different attention mechanisms underlie these two phenomena? Can a unifying explanation account for both? We conducted two experiments to decompose processing into component parts, and compared results to competing models' predictions. Mirror image search was unimpaired after target discrimination had been balanced across search conditions-results were consistent with an unlimited-capacity, decision noise model. Search asymmetry affected higher-level processing, however, resulting in capacity limitations that necessitated serial processing. A unifying explanation can account for these two seemingly unrelated phenomena. PMID- 16376403 TI - Short- and long-range spatial interactions: a redefinition. AB - First level short- and long-range spatial interactions are considered to be processed in the primary visual cortex. In psychophysics, they are measured with two kinds of stimuli, Gabor patches and lines/points. Each has its own short- and long-range definitions. We show that first, in terms of visual angle separation, the two definitions do not correspond to identical scales of interactions and second, that Gabor data can be matched to the lines/points definition by properly considering the observed effects. As a consequence, three regimes of spatial interaction are present: a case where overlapping of stimuli is present, and two others for spatially separated stimuli which we define as the short- and long range regimes. Both types of stimuli show compatible lateral interactions and, we think, permit the measurement of the same underlying mechanisms. PMID- 16376404 TI - Color blindness and contrast perception in cuttlefish (Sepia officinalis) determined by a visual sensorimotor assay. AB - We tested color perception based upon a robust behavioral response in which cuttlefish (Sepia officinalis) respond to visual stimuli (a black and white checkerboard) with a quantifiable, neurally controlled motor response (a body pattern). In the first experiment, we created 16 checkerboard substrates in which 16 grey shades (from white to black) were paired with one green shade (matched to the maximum absorption wavelength of S. officinalis' sole visual pigment, 492 nm), assuming that one of the grey shades would give a similar achromatic signal to the tested green. In the second experiment, we created a checkerboard using one blue and one yellow shade whose intensities were matched to the cuttlefish's visual system. In both assays it was tested whether cuttlefish would show disruptive coloration on these checkerboards, indicating their ability to distinguish checkers based solely on wavelength (i.e., color). Here, we show clearly that cuttlefish must be color blind, as they showed non-disruptive coloration on the checkerboards whose color intensities were matched to the Sepia visual system, suggesting that the substrates appeared to their eyes as uniform backgrounds. Furthermore, we show that cuttlefish are able to perceive objects in their background that differ in contrast by approximately 15%. This study adds support to previous reports that S. officinalis is color blind, yet the question of how cuttlefish achieve "color-blind camouflage" in chromatically rich environments still remains. PMID- 16376405 TI - Size distribution of wastewater COD fractions as an index for biodegradability. AB - The study proposes direct particle size measurement by sequential filtration and ultrafiltration as a convenient method for wastewater characterization for appropriate treatment technology. It also explores the correlation between particle size distribution (PSD) and chemical oxygen demand (COD) fractionation, as an index for biological treatability. Profiles obtained through PSD-based COD fractionation serve as the fingerprints for wastewaters, and as demonstrated in this study, reflect different pictures for textile wastewater and domestic sewage. PSD-based COD fractionation profiles identify the soluble range below 2 nm as the size interval housing both the soluble inert COD initially present in the wastewater and soluble inert microbial products generated during biological treatment, as also supported by the metabolic fractionation attained through respirometric analyses. Moreover, PSD-based color profiling offers a good index for the fate of biologically resistant chemicals passing through biological treatment. Compatible results obtained from comparative evaluation of PSD-based COD and color profiles provide useful information on the biodegradability of the textile wastewater studied. PMID- 16376406 TI - Linking catchment characteristics and water chemistry with the ecological status of Irish rivers. AB - Requirements of the EU Water Framework Directive for the introduction of ecological quality objectives for surface waters and the stipulation that all surface waters in the EU must be of 'good' ecological status by 2015 necessitate a quantitative understanding of the linkages among catchment attributes, water chemistry and the ecological status of aquatic ecosystems. Analysis of lotic ecological status, as indicated by an established biotic index based primarily on benthic macroinvertebrate community structure, of 797 hydrologically independent river sites located throughout Ireland showed highly significant inverse associations between the ecological status of rivers and measures of catchment urbanisation and agricultural intensity, densities of humans and cattle and chemical indicators of water quality. Stepwise logistic regression suggested that urbanisation, arable farming and extent of pasturelands are the principal factors impacting on the ecological status of streams and rivers in Ireland and that the likelihood of a river site complying with the demands of the EU Water Framework Directive, and be of 'good' ecological status, can be predicted with reasonable accuracy using simple models that utilise either widely available landcover data or chemical monitoring data. Non-linear landcover and chemical 'thresholds' derived from these models provide a useful tool in the management of risk in catchments, and suggest strongly that more careful planning of land use in Ireland is essential in order to restore and maintain water quality as required by the Directive. PMID- 16376407 TI - Halogenated acetaldehydes: analysis, stability and fate in drinking water. AB - In our previous studies, chloral hydrate has been the only chlorinated acetaldehyde determined in drinking water because authentic standards of other related haloacetaldehydes were not available. Recently, standards of dichloroacetaldehyde, bromochloroacetaldehyde, dibromoacetaldehyde, bromodichloroacetaldehyde, chlorodibromoacetaldehyde, and tribromoacetaldehyde have become available commercially. They were obtained and verified for purity and stability using a dual-column GC-ECD system. Each commercial standard was found to contain small amounts of the other target haloacetaldehydes (HAs). The stability of the HAs stock solutions was solvent dependent: in acetone, the brominated species partially degraded to bromoacetone, while all target HAs were stable in MTBE for up to 8 months. The analytical parameters, required for the quantification of HAs in water, were determined and used to evaluate the stability of the HAs in water. Under the conditions of the sampling protocol (field pH adjustment to pH 4.5 and storage at 4 degrees C), the target HAs were stable in water for up to 14 days. However, at typical drinking water pH and temperature conditions, the stability varied with the HA species, pH, temperature and storage period. The trihalogenated acetaldehydes degraded, in part, to their corresponding trihalomethanes (THMs) at increasing pH and temperature. Most target HAs were detected in drinking water samples collected from various Canadian drinking water systems, and the speciation was dependent on water parameters (e.g. bromide concentration) and treatment processes. From the water samples analysed, chloral hydrate ranged between 7% and 51% of the total HAs (w/w). The weigh ratio of total HAs to total THMs (10-46%) indicated that HAs contributed significantly to the pool of DBPs in drinking water. PMID- 16376408 TI - Effect of phosphate addition on the sorption-desorption reaction of selenium in Japanese agricultural soils. AB - Desorption levels of soil-sorbed selenium (Se) were studied by adding phosphate to 22 typical Japanese agricultural soils. Soil-soil solution distribution coefficients of Se (Kd-Se) were measured using a batch process as an index of Se sorption level, adding 75Se as a tracer. After the Kd measurement, extraction of soil-sorbed 75Se with a 0.1 M or 1 M Na2HPO4 solution followed to determine the amount of 75Se desorbed by the phosphate. When the 0.1 M Na2HPO4 solution was used, 18-70% of soil-sorbed Se was extracted (average: 47%). However, when the 1 M Na2HPO4 solution was used, 27-83% of soil-sorbed Se was extracted (average: 57%). The observed 75Se desorption percentage indicated the maximum Se removability by phosphate addition. The desorption percentage of Se with 1 M Na2HPO4 correlated with Kd-Se values, suggesting that the soil sample with higher Kd-Se contained more reactive components for phosphate-sorption than the soil sample with lower Kd-Se. To evaluate the effect of phosphate concentration on the Se sorption, the Kd-Se was measured for two typical soils under different levels of phosphate (0.1-10 mM PO4). The Kd values were decreased by phosphate addition for both soils. The Kd decrease was observed even for just 1 mM PO4. The phosphate addition with 1 mM PO4 is the same level as in P fertilizer applied to paddy fields in Japan. Therefore, it was suggested that Se desorption should occur in Japanese soils due to the phosphate input. PMID- 16376409 TI - Effect of calcite on Pb-doped solidified waste forms in leaching. AB - The alkalinity of cementitious materials rectifies a low pH of leachant to be over 12 in leaching. The rapid change of leachant pH produces the remarked variation of solubility of heavy metals in the toxicity characteristic leaching procedure. The release of lead on cementitious solid waste forms in leaching was observed in the pH range of particularly 12. The release of lead is significant dependant on the changed values of leachant pH. The pH static leaching procedure (PSLP) was implemented for assessing a quantitative measurement of total potential leachability with the continuing addition of acidic leachant up to the point of no change of leachant pH. The release of lead on Pb-doped solidified waste forms (SWF), in the PSLP, was 27.0% of initial concentration at the maximum meq CH3COOH (about 24.0) added to g dry solidified wastes. In this study, the immobilization effect of calcite on SWF was investigated in the acidic condition by the PSLP. Calcite additions to SWF make a good fixation efficiency of about 92%. Calcite addition abundantly provides the acid neutralization capacity to protect SWF from the attack of acid, with the marked development of hydrated minerals (mainly portlandite and C-S-H) in accordance with the accelerating effect of cement hydration. PMID- 16376410 TI - Limits on tongue deformation--Diana monkey formants and the impossible vocal tract shapes proposed by Riede et al. (2005). PMID- 16376411 TI - Of muscle-bound crania and human brain evolution: the story behind the MYH16 headlines. PMID- 16376413 TI - Locomotor behavior and long bone morphology in individual free-ranging chimpanzees. AB - We combine structural limb data and behavioral data for free-ranging chimpanzees from Tai (Ivory Coast) and Mahale National Parks (Tanzania) to begin to consider the relationship between individual variation in locomotor activity and morphology. Femoral and humeral cross sections of ten individuals were acquired via computed tomography. Locomotor profiles of seven individuals were constructed from 3387 instantaneous time-point observations (87.4 hours). Within the limited number of suitable chimpanzees, individual variation in locomotor profiles displayed neither clear nor consistent trends with diaphyseal cross-sectional shapes. The percentages of specific locomotor modes did not relate well to diaphyseal shapes since neither infrequent nor frequent locomotor modes varied consistently with shapes. The percentage of arboreal locomotion, rather than estimated body mass, apparently had comparatively greater biological relevance to variation in diaphyseal shape. The mechanical consequences of locomotor modes on femoral and humeral diaphyseal shapes (e.g., orientation of bending strains) may overlap between naturalistic modes more than currently is recognized. Alternatively, diaphyseal shape may be unresponsive to mechanical demands of these specific locomotor modes. More data are needed in order to discern between these possibilities. Increasing the sample to include additional free-ranging chimpanzees, or primates in general, as well as devoting more attention to the mechanics of a greater variety of naturalistic locomotor modes would be fruitful to understanding the behavioral basis of diaphyseal shapes. PMID- 16376414 TI - Electrodialytic remediation of CCA-treated waste wood in a 2 m(3) pilot plant. AB - Waste wood that has been treated with chromated-copper-arsenate (CCA) poses a potential environmental problem due to the content of copper, chromium and arsenic. A pilot plant for electrodialytic remediation of up to 2 m(3) wood has been designed and tested and the results are presented here. Several process parameters were investigated, and it was found that the use of collecting units and soaking of the wood prior to the electrodialytic process had a positive influence on the remediation process. There was a tendency towards higher removal of CCA from wood chips <2 cm, compared to larger wood size fractions. The best remediation efficiency was obtained in an experiment with an electrode distance of 60 cm, and 100 kg wood chips. In this experiment 87% copper, 81% chromium and >95% arsenic were removed. One other experiment was also analysed for arsenic. In this experiment the distance between the working electrodes was 1.5 m and here 95% As was removed. The results showed that arsenic may be the easiest removable of the copper, chromium and arsenic investigated here. This is very encouraging since arsenic is the CCA components of most environmental concern. PMID- 16376417 TI - Quality of life (QOL) outcomes from a randomized trial of cisplatin versus cisplatin plus paclitaxel in advanced cervical cancer: a Gynecologic Oncology Group study. AB - OBJECTIVE: Chemotherapy can profoundly affect patients' quality of life (QOL), yet few clinical trials in advanced cervical cancer have included QOL outcomes. Our purpose was to assess the impact of cisplatin (C) versus cisplatin plus paclitaxel (CP) on overall QOL and pain in cervical cancer patients. METHODS: QOL was assessed using FACT-Cx, consisting of the Functional Assessment of Cancer Therapy (FACT-G) plus a cervix cancer-specific subscale, the Brief Pain Inventory Short Form (BPI-SF), and a neurotoxicity subscale. Time points were: baseline (prior to randomization) and prior to chemotherapy cycles 2, 3, and 4. RESULTS: Overall (FACT-G) scores did not differ significantly between arms at the fourth assessment (C = 70.3 (19.6); CP = 72.8 (17.4)). Scores were stable over time and considerably lower than the general population norms. The BPI-SF revealed a decline in pain scores in both arms from the first to fourth assessments. The CP arm produced a significantly higher response rate and progression-free survival (PFS) but not overall survival (OS). Greater myelosuppression was reported in the combination arm. The rate of QOL drop-out for any reason was higher for C (53%) compared to CP (38%) (P < 0.05). At the fourth time point, 60% of living patients in both arms completed a QOL assessment. CONCLUSION: There was no significant difference in overall QOL scores between treatment arms or serially. Combined with QOL results, the significant increase in response and PFS in the CP arm and the higher drop-out rate in the C arm suggest a better outcome for the combination regimen despite its increased myelosuppression. PMID- 16376418 TI - SLE and CA-125. PMID- 16376419 TI - Re: "Gemcitabine and liposomal doxorubicin in the salvage treatment of ovarian cancer: updated results and long-term survival". PMID- 16376421 TI - The influence of environmental conditions on the material properties of setting glass-ionomer cements. AB - OBJECTIVES: Aim of this study was to investigate the influence of temperature on the setting time and compressive strength of two conventional glass-ionomer cements (GIC's) and to determine the influence of storage medium, oil or water and storage time. MATERIALS AND METHODS: Two conventional GIC's, Ketac Molar (3 M ESPE Dental Products, Seefeld, Germany) and Fuji IX Fast (GC Corp., Tokyo, Japan) were used to perform flow property tests and compression tests. Flow property measurements were performed using a displacement rheometer at six different temperatures. From the results of the rheometer tests, the working times and setting times could be determined. The samples for the compressive tests were stored at four different temperatures and in two different media. Testing took place at five time intervals reaching from 1 h to 3 months. RESULTS: The results of rheometer tests showed that a temperature increase speeded up the setting reaction significantly. The compressive strength results showed a jump in time as a result of the higher curing temperature but no long-term strength effect was observed. Materials curing in oil reached a significantly higher compressive strength compared to storage in water and Fuji IX Fast is significantly stronger than Ketac Molar. SIGNIFICANCE: It was concluded that a temperature between 333 and 343 K almost sets conventional GIC's on command and improves the early compressive strength. PMID- 16376420 TI - Estrogenic encounters: how interactions between aromatase and the environment modulate aggression. AB - Initial investigations into the mechanistic basis of aggression focused on the role of testosterone (T) and a variety of studies on non-human animals found that elevated T levels promote aggression. However, many correlational studies have not detected a significant association between aggression and peripheral T levels. One reason for this inconsistency may be due to differential metabolism of T within the brain, in particular, the conversion of T to estrogen by aromatase. Thus, differences in aromatase enzyme activity, estrogen receptor expression, and related cofactors may have important effects on how steroids affect aggressive behavior. Hormone manipulation studies conducted in a wide variety of species indicate that estrogens modulate aggression. There is also growing evidence that social experience has important effects on the production of estrogen within the brain, and some cases can not be explained by androgenic regulation of aromatase. Such changes in central aromatase activity may play an important role in determining how social experiences affect the probability of whether an individual engages in aggressive behavior. Although studies have been conducted in many taxa, there has been relatively little integration between literatures examining aggression in different species. In this review, we compare and contrast studies examining aggression in birds, mammals, and humans. By taking an integrative approach to our review, we consider mechanisms that could explain species differences in how estrogen modulates aggression. PMID- 16376422 TI - Network structure of Bis-GMA- and UDMA-based resin systems. AB - OBJECTIVES: The commonly used dental base monomers 2,2-bis[p-(2'-hydroxy-3' methacryloxypropoxy)phenylene]propane (Bis-GMA) and 1,6-bis(methacryloxy-2 ethoxycarbonylamino)-2,4,4-trimethylhexane (UDMA) require the use of a diluent monomer, such as triethylene glycol dimethacrylate (TEGDMA). The aim of this study was to measure double bond conversion of UDMA/TEGDMA and Bis-GMA/TEGDMA polymeric systems, determine the leachable portion, and analyze network formation by evaluating crosslinking and pendant double bonds. METHODS: UDMA or Bis-GMA was combined with TEGDMA in systematic increments and irradiated to form light cured polymers. Fourier transform infrared spectroscopy in the near-infrared region was used to measure double bond conversion. The leachable sol fraction was analyzed by 1H NMR. Resin composites were formulated. Flexural strength was measured by three-point bending and volumetric shrinkage was determined with a mercury dilatometer. RESULTS: The amount of base monomer greatly influenced double bond conversion, sol fraction, and crosslinking. Increasing base monomer concentration decreased double bond conversion, increased the leachable fraction, and decreased crosslinking and network formation. At mole fractions higher than 0.125, the UDMA polymers had significantly higher conversion than the Bis-GMA polymers. Bis-GMA polymers had higher leachable amounts of unreacted monomer, while UDMA mixtures had more crosslinking than the Bis-GMA mixtures. In regards to the physical properties of resin composites, increasing the base monomer improved flexural strength and decreased volumetric shrinkage. SIGNIFICANCE: This systematic study for the evaluation of conversion, leachability, crosslinking, and network structure along with physical properties, like volumetric shrinkage and flexural strength, are required for the optimization of competing desirable properties for the development of durable materials. PMID- 16376423 TI - Membrane-mimetic films containing thrombomodulin and heparin inhibit tissue factor-induced thrombin generation in a flow model. AB - Membrane-mimetic thin films containing thrombomodulin (TM) and/or heparin were produced and their capacity to inhibit thrombin generation evaluated in a continuous flow system. Tissue factor (TF) along with TM and heparin were immobilized in spatially restricted zones as components of a membrane-mimetic film. Specifically, TF was positioned as an upstream trigger for thrombin generation and TM and/or heparin positioned over the remaining downstream portion of test films. Peak and steady-state levels of thrombin were decreased by antithrombin III (ATIII), as well as by surface bound heparin and TM. Although physiologic concentrations of ATIII have the capacity to significantly inhibit thrombin activity, surface bound TM and heparin nearly abolished steady-state thrombin responses. In particular, surface bound TM appears to be superior to heparin in reducing local thrombin concentrations. These studies are the first to demonstrate the additive effect of surface bound heparin and TM as a combined interactive strategy to limit TF-induced thrombin formation. PMID- 16376425 TI - Ghrelin is expressed in trigeminal neurons of female mice in phase with the estrous cycle. AB - Several disorders mediated by the trigeminal nerve including migraine and temporomandibular disorder (TMD) are more common in women than in men, and painful attacks are often linked to the menstrual cycle. Estrogen receptors in trigeminal neurons may be involved in regulating neuronal function, causing changes in sensitivity that contribute to these attacks. In a previous study, we demonstrated that expression of specific neuropeptides including galanin and neuropeptide Y in trigeminal ganglia of female rodents varies with the estrous cycle. In this study, we examined expression of the orexigenic peptide ghrelin in trigeminal ganglia of cycling female mice. RT-PCR studies demonstrated that ghrelin mRNA is upregulated by over 5-fold at the high estrogen stages of the cycle, proestrus and early estrus over the levels expressed at the low estrogen stage of the cycle, diestrus. Double-labeling immunohistochemical studies and cell size measurements were conducted to identify the phenotype of neurons in trigeminal ganglia containing ghrelin. Ghrelin was present in trigeminal neurons containing peripherin, a marker of neurons with unmyelinated axons, in trigeminal neurons binding IB4, a marker of nonpeptidergic nociceptors, in trigeminal neurons containing neurofilament H, a marker of neurons with myelinated axons, and in trigeminal neurons containing the neuropeptide calcitonin gene-related peptide (CGRP). Ghrelin-positive neurons averaged 25.6 microm in diameter, but included neurons in all the size ranges except the smallest peripherin-positive neurons. Thus, nearly all of the major populations of trigeminal neurons including peptidergic and nonpeptidergic nociceptors contain ghrelin. These studies suggest that ghrelin, a multifunctional peptide, may contribute to the mechanism linking orofacial pain syndromes in females, including temporomandibular disorder and migraine, to cyclical hormonal changes. PMID- 16376424 TI - ICAM-1 is overexpressed by villous trophoblasts in placentitis. AB - Although an in vitro study has hypothesized that expression of ICAM-1 by villous trophoblasts could be important for the influx of maternal immune cells in villitis, it remains to be shown whether the same phenomenon occurs in human villitis. To investigate the expression of ICAM-1 by villous trophoblasts, its relationship with rupture of the trophoblastic barrier and influx of immune cells into the villi, we analysed 18 paraffin-embedded placentas with placentitis (5 by Toxoplasma gondii, 3 by Trypanosoma cruzi, 2 by Paracoccidioides brasiliensis and 8 of unknown aetiology - VUE) and 8 control placentas for detection of ICAM-1 by immunohistochemistry. All cases but one of placentitis showed trophoblast overexpression of ICAM-1 in the inflamed villi, located almost exclusively next to the areas of trophoblastic rupture. The villitis cases (caused by T. cruzi, T. gondii and VUE) presented leukocyte adherence in the areas of trophoblastic rupture. When the inflammatory reaction was situated in the intervillous space (placentitis by P. brasiliensis), in spite of the trophoblastic rupture and ICAM 1 overexpression there was no leukocyte influx into villi. None of the control placentas showed ICAM-1 expression by the trophoblast. We concluded that overexpression of ICAM-1 by villous trophoblasts occurs during placentitis characterized by accumulation of leukocytes in the villous or intervillous space and probably plays an important role in the rupture of the trophoblastic barrier. The influx of immune cells into the villi appears to be mediated by ICAM-1 but the location of the antigen within villous stroma is certainly a crucial factor for its occurrence. PMID- 16376426 TI - Accumulation and elimination of cadmium in larval stage zebrafish following acute exposure. AB - A number of recent studies have examined the impact of acute cadmium exposure on early zebrafish development at the morphological, cellular, and molecular levels. However, no information on the accumulation and elimination of cadmium during early life stages of zebrafish development has been available. Here we have quantified cadmium accumulation in larval zebrafish (Danio rerio) by graphite furnace atomic absorption spectroscopy following short-term acute exposure and recovery periods. Zebrafish (80 h postfertilization) were exposed to various concentrations of cadmium (0.2, 1.0, 5.0, 25, 125 microM) for 3 h. Cadmium accumulation in larvae increased with exposure concentration. After exposure at 5.0, 25, and 125 microM cadmium, the fish were allowed to recover in freshwater for 0, 12, or 24 h. Cadmium content did not show a statistically significant decrease over the recovery period when exposed to 5.0 or 25 microM cadmium, whereas significant losses over the recovery period were observed following 125 microM exposure. These results suggest that the larval zebrafish decrease total cadmium body burden only following relatively high short-term acutely toxic exposures. PMID- 16376427 TI - The usage of alternative splice sites in Mus musculus synaptotagmin-like 2 gene is modulated by cyclosporin A and FK506 in T-lymphocytes. AB - Cyclosporin-A and FK506 block the calcineurin activity preventing the transcription of genes sharing NFAT-like binding sequences in their promoter region. We presently show that activation of murine T-cells in presence of these immunosuppressors results in the up-regulation of the synaptotagmin-like 2 gene. However, of the four known isoforms, only mRNAs encoding the a and b isoforms accumulate. Two previously undected isoforms, each characterized by the retention of an intron, were found. The first, Slp2-e, includes exon 8, intron 8 and exon 9. The second, Slp2-f, is composed of exon 7, intron 7 and exon 8. Slp2-f has an open reading frame coding for a putative protein of 1229 amino acids sharing 47% identities with the human breast-associated antigen, SGA-72 M. In addition to the well-documented modulation of gene transcription, the two immunosuppressors also play a role in the choice of alternative splice sites on murine Slp2 pre-mRNA. PMID- 16376428 TI - A rapid method for mass screening for parkinsonism. AB - Epidemiology studies of parkinsonism employ a variety of techniques for unbiased sampling of populations. No current method permits mass screening of all subjects in a population for parkinsonism by movement disorders specialists. We developed and piloted a new approach to facilitate accurate and efficient screening of large populations for diagnosis of parkinsonism and provide data on sensitivity and specificity. We evaluated 2081 welders referred for medical-legal screening. Subjects were video taped using a standardized protocol, and videos were rated on the Unified Parkinson's Disease Rating Scale motor subsection 3 (UPDRS3). A "video rater" viewed video tapes and entered ratings through a web-based database. An "in-person" examiner performed a UPDRS3 examination in a randomly selected subgroup of 48 workers drawn from the 2081. We developed quantitative diagnostic criteria for parkinsonism that established minimum diagnostic thresholds based upon UPDRS3 scores and compared these criteria with diagnosis by an in-person examiner. Specificity of these criteria compared to in-person examination was 91-100% but sensitivity was 56%. A threshold UPDRS3 score greater than nine provided 100% sensitivity and 81% specificity. Liberal criteria identified 266 (13.1%) subjects with probable parkinsonism and 220 (10.8%) subjects with definite parkinsonism. Conservative criteria identified 260 (12.8%) with probable parkinsonism and 122 (6%) with definite parkinsonism. Our screening method permits rapid assessment of parkinsonian signs. An absolute UPDRS3 score greater than nine provided the best combination of sensitivity and specificity for the diagnosis of parkinsonism, while quantitative exam-based criteria for cardinal parkinsonian signs maximized specificity. Parkinsonism as diagnosed by our criteria was common in this group of welders. PMID- 16376429 TI - The binding of Ni(II) ions to hexahistidine as a model system of the interaction between nickel and His-tagged proteins. AB - The aim of this work is to study the binding of nickel ions to hexahistidine (His(6)) combining potentiometric titrations and spectroscopic (UV-Vis and circular dichroism) determinations in order to establish the species distribution as a function of the pH, their stoichiometry, stability and geometry. For comparative purposes, the same procedure was applied to the Ni-histidine (His) system. His behaves as a tridentate ligand, coordinating the carboxyl group, the imidazole and the amino nitrogen atoms to Ni(II) ions in an octahedral coordination and a bis(histidine) complex is formed at pH higher than 5. For the Ni-His(6) system, the complex formation starts at pH 4 and five different species (Ni(His(6))H, Ni(His(6)), Ni(n)(His(6))(n), Ni(n)(His(6))(n)H(-n/2), Ni(n)(His(6))(n)H(-n)) are formed as a function of the pH. Ni(His(6))H involves the coordination of the imidazole nitrogen and a deprotonated amide nitrogen (N(Im), N(-)) resulting in an octahedral geometry. In Ni(His(6)), an imidazole nitrogen is deprotonated and coordinated (2N(Im), N(-)) to the metal ion with a square planar geometry. The aggregated forms result from the extra Ni-N(Im) coordination, resulting in a 4N square planar geometry that is stabilized by inter/intramolecular hydrogen bonds. This coordination mode is not altered during the deprotonation steps from Ni(n)(His(6))(n). PMID- 16376430 TI - A quantitative method for comparison of expression of alternatively spliced genes using different primer pairs. AB - In this paper we describe a novel two-step method for comparison of expression of alternatively spliced genes by quantitative PCR (QPCR) applying different primer pairs. As a model system we used rat decay accelerating factor (DAF; CD55) mRNA, which comprises three different isoforms: soluble (sDAF), transmembrane (tmDAF) and glycosyl-phosphatidylinositol (GPI) anchored (gpiDAF) forms. The first step was to prepare solid phase specific for each mRNA isoform and purify the three DAF-forms from total RNA. We then assessed amplification efficiency of primer pairs designed to recognise each of the isoforms using equimolar amounts of the three purified DAF mRNAs. The final step in our assay was to compare expression of the three DAF-isoforms in testis by QPCR taking into account the efficiency of their amplification to enable quantification. The RNA capture/QPCR method we described here can be used for quantifying the expression ratios of alternatively spliced mRNAs from a single gene or for direct comparison of expression of different genes. PMID- 16376431 TI - Symptomatic and functional outcome of bipolar disorder in Butajira, Ethiopia. AB - BACKGROUND: Limited information is available on the outcome of bipolar disorder in developing countries. OBJECTIVE: To describe the symptomatic and functional outcome of bipolar disorder. METHODS: The psychoses and affective disorder modules of the CIDI were used to screen 68,378 individuals by a door-to-door survey of a defined district in Ethiopia. In addition, key informants were used to identify individuals with probable major mental illnesses. SCAN interviews were completed at the second stage to confirm the diagnosis. A total of 315 cases of bipolar disorder were identified, of which 264 (69 recent-onset and 195 prevalent cases) were prospectively followed for a mean of 2.5 (range 1-4) years by baseline and annual clinical assessments using symptom rating scales. Functional dimensions of the SF-36 scale were used to describe functional outcome. Random coefficient analyses were used to evaluate potential correlates of outcome. RESULTS: The magnitudes of mania and depression symptoms were elevated at baseline but improved with follow-up, although the improvement was less marked for depression. Sociodemographic or clinical variables were not associated with the improvements in symptomatic outcome. Between 35% and 47% of the recent-onset cases had functional role restrictions, while 42-52% of long standing cases had such restrictions during the follow-up years. Similarly, social and physical functioning deficits were also present in 52-86% and 35-47% of recent-onset and long-standing cases, respectively. The magnitude of depression and mania symptoms was associated with poor functional outcome, while male sex, rural residence and being married were associated with better functional outcome. CONCLUSION: Although there were improvements in function with follow-up, between one-third and one-half of cases continued to have functional deficits. PMID- 16376432 TI - Influence of episode duration of major depressive disorder on response to electroconvulsive therapy. AB - BACKGROUND: Longer duration of major depressive episode is supposed to decrease response to electroconvulsive therapy (ECT). Most studies on the subject are dated and their population differs from ours, therefore their results may not be applicable to our population of severely depressed inpatients. METHODS: We reviewed the records of 56 consecutive inpatients with major depressive disorder according to DSM-III-R criteria and assessed each patient's episode duration. We examined whether episode duration has an effect on response to ECT. RESULTS: Episode duration has no significant effect on response to ECT, according to both a reduction on the Hamilton Rating Scale for Depression (HRSD) of at least 50% and a post-treatment HRSD score 100 other normal serum samples tested, indicating a very low frequency in the general population. The low affinity of these unusually present antibodies could explain the absence of disease, despite their relatively high titer. The significance of this finding in the origin of disease-associated anti-GM1 antibodies is discussed. PMID- 16376438 TI - Holt-Oram syndrome presenting as agenesis of the left pericardium. AB - We report on a case of a 60-year-old man with progressive heart failure, mitral and aortic valve insufficiency and bilateral asymmetrical skeletal upper-limb deformities. Central to the suspicion of Holt-Oram syndrome in this patient was the surgical finding of agenesis of the left pericardium. A Holt-Oram syndrome diagnostic was confirmed through molecular analysis of the TBX5 gene. A new amino acid substitution at position 61 of the TBX5 gene was identified and confirmed the clinical diagnosis of Holt-Oram syndrome. The clinical presentation of the present case broadens the clinical spectrum of Holt-Oram syndrome and point out the importance of Tbx 5 in pericardium development. It is still an unstudied issue whether TBX5 mutations may also be present in other clinical presentations where absence of the pericardium is a feature. PMID- 16376439 TI - Evolution of aortic valve replacement in children: a single center experience. AB - BACKGROUND: The availability of an ideal prosthesis for aortic valve replacement (AVR) in children remains controversial due to an early degeneration of xenografts and the potential risks related to anticoagulation with mechanical prostheses. This has led many surgeons to the Ross procedure. This study outlines the evolution of our 30-year experience with AVR in children. METHODS: One hundred and forty-seven children, aged 10 days to 18 years (mean 11.9+/-5.7 years), underwent AVR between 1974 and June 2005. Preoperative diagnosis included aortic insufficiency (n=39), aortic stenosis (n=14), combined aortic stenosis with insufficiency (n=78), and complex left ventricular outflow tract obstruction (n=16). Xenografts (n=11) and mechanical prostheses (n=47) were used in 58 patients. The remaining 89 patients had placement of homografts (n=8) or underwent a Ross procedure (n=81). Of the 147 patients, 87 (59%) had previous procedures. RESULTS: Overall early and late mortality was 7.5% (11/147 pts). Overall survival estimated by the Kaplan-Meier method, including early mortality, was 94% at 1 year, and 93% at 5, 10, and 20 years. Univariate and multivariate analysis identified date of operation (before 1980) as a risk factor for death (p=0.002). Follow-up was complete in 136 patients (5 lost to follow-up), with a total follow-up of 2433.72 patient-years. The overall study group aortic valve related reoperation rate was 20% (20/138 pts), the reoperation rate was highest in xenograft group (60%); followed by mechanical valves group (16%), homograft group (14%), and the Ross procedure group (9%). Ross patients showed significant increase of the annulus diameter (p=0.002) and the aortic sinus diameter (p=0.01) at the last follow-up. The actuarial rate for freedom from aortic valve-related reoperation was 99% at 1 year, 94% at 5 years, 88% at 10 years, and 85% at 20 years. Univariate and multivariate analysis identified the presence of a xenograft as a risk factor for aortic valve-related reoperation (p=0.001). CONCLUSION: AVR in children can be performed with acceptable mortality and minimal mid-term morbidity. The Ross procedure, although more complicated, has the advantage of not requiring anticoagulation. Pulmonary autograft, in our series, has demonstrated growth with no structural degeneration. The potential for development of significant autograft insufficiency and ascending aortic aneurysmal dilatation is small but warrants annual follow-up. Our data supports that the Ross procedure is the AVR of choice in children. PMID- 16376440 TI - Dysautonomia and ventricular dysfunction in the indeterminate form of Chagas disease. AB - BACKGROUND: The associations between autonomic function and biventricular function in patients with the indeterminate form of Chagas disease remains to be elucidated. METHODS: In 42 asymptomatic patients and 19 healthy volunteers, the autonomic function was assessed by time domain indices of heart rate variability (HRV), analyzed for 24 h; the right ventricular function was assessed by fraction area change, right ventricle shortening, and systolic excursion of the tricuspid valve; and the left ventricular function was assessed by ejection fraction and transmitral flow velocities. Data were expressed as mean+/-SD or medians (including the lower quartile and upper quartile). Groups were compared by Student's t or Mann-Whitney U test. Autonomic and ventricular function were correlated by Pearson's or Spearman's correlation coefficient. The level of significance was 5%. RESULTS: Right and left ventricular systolic function indexes were comparable between groups. Transmitral flow velocities were decreased in the Chagas disease group (p<0.05). The patients presented impaired HRV as indicated by the values of SDNN-day (80 (64-99) ms vs. 98 (78-127) ms; p=0.045), SDNNI-24 h (54 (43-71) vs. 65 (54-105) ms; p=0.027), SDNNI-day (49 (42 64) vs. 67 (48-76) ms; p=0.045), pNN50-day (2.2 (0.7-5)% vs. 10 (3-11)%; p=0.033); and pNN50-24 h (3 (1-7)% vs. 12 (8-19)%; p=0.013). There were no correlations between the left ventricular diastolic indices and autonomic dysfunctional indices (p>0.05). CONCLUSION: Patients with the indeterminate form of Chagas disease have both dysautonomia and left ventricular diastolic dysfunction. However, the right ventricular function is preserved. Importantly, ventricular diastolic dysfunction and dysautonomia are independent phenomena. PMID- 16376441 TI - The future of adult patients after Mustard or Senning repair for transposition of the great arteries. AB - BACKGROUND: After Mustard or Senning repair for transposition of the great arteries (d-TGA), qualitative assessment of the systemic right ventricle remains difficult. We wanted to evaluate the relationship between demographic and echocardiographic variables, and exercise performance. METHODS: In 22 consecutive d-TGA patients (9 Mustard and 13 Senning) a standard transthoracic echocardiogram was performed. Peak oxygen consumption (peak VO2, ml/kg/min) was obtained by a bicycle stress test. Cardiac index (CI) at rest, 50 W, and 100 W was calculated from cardiac output, obtained by CO2 rebreathing. RESULTS: The group of patients consisted in 15 male and 7 female with a median age of 21 (range 17-34) years. On echocardiographic evaluation, right ventricular function was normal in one, slightly impaired in 11, moderately impaired in 9, and severely impaired in one. Peak VO2 was 27.4+/-7.9 ml/kg/min, which was 64+/-16% of predicted values in normals. CI at rest, 50 Watt, and 100 Watt, were 2.7+/-0.5, 5.6+/-1.2, and 6.8+/ 1.1 l/min/m2, respectively. No relationship between echocardiographic parameters and exercise capacity was found. Age correlated significantly with CI at 50 and 100 W (rho=-0.44, P=0.045 and rho=-0.77, P=0.0001, respectively). Finally, simple regression analysis identified a linear relationship between age and CI at 100 W (beta=-0.54 and R2=0.29, P=0.014). CONCLUSIONS: Maximal exercise capacity is reduced in adult patients after Mustard or Senning repair. Standard echocardiographic 2D-measurements are insufficient to determine the reduction in their functional capacity. The inverse relationship between age and the cardiac indices suggests an age dependent and progressive diminution of systemic ventricular function. PMID- 16376442 TI - Soluble CD40 ligand and interleukin-6 in the coronary circulation after acute myocardial infarction. AB - BACKGROUND: Inflammation, operated by blood, vascular and immune cells interaction, is implicated in plaque disruption and CD40 ligand (CD40L) was identified on activated T cells and platelets. We sought to investigate the roles of local inflammation in acute myocardial infarction (AMI). METHODS: Coronary sinus (CS) and arterial (A) levels of interleukin (IL)-6 and soluble CD40L (sCD40L) and matrix metalloproteinase (MMP)-9 activity in serial blood samples obtained until 48 h after percutaneous coronary intervention (PCI) were determined. In tissue specimens obtained by aspirating thrombectomy and directional coronary atherectomy, CD40L was immunohistochemically stained. RESULTS: Trans-cardiac gradient (CS-A) of IL-6, indicating cardiac release into the coronary circulation, significantly increased at 24 h after PCI in patients with AMI (group MI, n=17) in contrast with angina pectoris (n=10). Soluble CD40L levels in CS showed earlier peak, yielding trans-cardiac gradient, at 9 h in both groups. The maximum (max) release of IL-6 in MI, but not sCD40L, positively correlated with end-diastolic volume index (R=0.84) and negatively with ejection fraction (R=-0.66) by contrast ventriculography at 6-month follow up. Immunohistological study revealed the expression of CD40L in intra-coronary occlusive and mural thrombi. Aspirating thrombectomy significantly reduced the increase in both sCD40L levels and MMP-9 activity, but not max IL-6 release in MI. CONCLUSIONS: In contrast with myocardial injury represented by IL-6 release, acute rise in sCD40L levels with the MMP-9 activation in the coronary circulation may possibly reflect local inflammation with platelet activation and be a novel marker of plaque damage by PCI. PMID- 16376443 TI - Severe cardiomyopathy associated to cocaine abuse. PMID- 16376444 TI - A technique of using gated-CT images to determine internal target volume (ITV) for fractionated stereotactic lung radiotherapy. AB - BACKGROUND AND PURPOSE: To develop and evaluate a technique and procedure of using gated-CT images in combination with PET image to determine the internal target volume (ITV), which could reduce the planning target volume (PTV) with adequate target coverage. PATIENTS AND METHODS: A skin marker-based gating system connected to a regular single slice CT scanner was used for this study. A motion phantom with adjustable motion amplitude was used to evaluate the CT gating system. Specifically, objects of various sizes/shapes, considered as virtual tumors, were placed on the phantom to evaluate the number of phases of gated images required to determine the ITV while taking into account tumor size, shape and motion. A procedure of using gated-CT and PET images to define ITV for patients was developed and was tested in patients enrolled in an IRB approved protocol. RESULTS: The CT gating system was capable of removing motion artifacts for target motion as large as 3-cm when it was gated at optimal phases. A phantom study showed that two gated-CT scans at the end of expiration and the end of inspiration would be sufficient to determine the ITV for tumor motion less than 1 cm, and another mid-phase scan would be required for tumors with 2-cm motion, especially for small tumors. For patients, the ITV encompassing visible tumors in all sets of gated-CT and regular spiral CT images seemed to be consistent with the target volume determined from PET images. PTV expanded from the ITV with a setup uncertainty margin had less volume than PTVs from spiral CT images with a 10-mm generalized margin or an individualized margin determined at fluoroscopy. CONCLUSIONS: A technique of determining the ITV using gated-CT images was developed and was clinically implemented successfully for fractionated stereotactic lung radiotherapy. PMID- 16376445 TI - The optimization of dose delivery for intraoperative high-dose-rate radiation therapy using curved HAM applicators. AB - BACKGROUND AND PURPOSE: To determine the effect of the curvature of Harrison Anderson-Mick applicators on the dose distribution in high-dose-rate intraoperative radiation therapy (HDR-IORT). MATERIAL AND METHODS: Treatment planning was performed with flat applicators using (192)Ir as the radioactive source, and dwell times were optimized using dose-point optimization techniques. These optimized dwell times were then used for the curved applicators, and the dose distributions that would actually be delivered to patients were determined. RESULTS: The dose directly below the central catheter was strongly dependent on the curvature of the applicator. Steep parabolic curves caused underdoses of as much as 19% at a point 1cm from the convex side of the applicator. The rate of dose reduction with increasing distance from the applicator surface was also a function of the curvature of the applicator. CONCLUSIONS: The curvature of the applicator profoundly affects dosimetry and can be exploited to optimize coverage of the target during HDR-IORT procedures. To ensure accurate dose delivery, these dose perturbations must be accounted for in the planning process. We recommend maintaining a dosimetry atlas of various applicator sizes and curvatures in addition to one for flat applicators. PMID- 16376447 TI - Distribution of airborne microorganisms in commercial pork slaughter processes. AB - The objective of this research was to determine the prevalence and distribution of airborne bacterial contamination, with particular reference to Escherichia coli and Salmonella, at a number of stages in a pork slaughtering plant. Air samples (impaction and sedimentation) were recovered from seven locations before and during operations in a commercial pork processing plant. Aerobic mesophilic bacteria, E. coli counts and the incidence of Salmonella in the air were determined. Most sample locations which provided high impaction counts also provided high sedimentation counts. Before commencement of operations, there were no significant differences in aerobic mesophilic bacteria obtained from the sample locations. However, within 2 h of the commencement of operations, aerobic mesophilic bacteria in the wet room (3.14 log10 cfu/m3) were significantly higher (P < 0.05) than those in the clean room (2.66 log10 cfu/m3) and chiller (2.34 log10 cfu/m3). By the afternoon, similar aerobic mesophilic bacteria counts were recovered in the wet and clean rooms, although counts in both of these areas were significantly higher (P > 0.05) than in the chiller. In general there were no significant differences in E. coli counts between rooms (wet room, clean room and chiller) and these did not increase during the production day. Salmonella were detected at the locations of the dehairing and evisceration operations. Aerobic mesophilic bacteria in the air within the abattoir increased as production proceeded. In addition the air within the abattoir contained organisms such as Salmonella and E. coli. Positive correlations (P < 0.05-P < 0.001) between impaction and sedimentation samples were found suggesting that air may be an important source of carcass contamination. PMID- 16376446 TI - Low-dose intraoperative brachytherapy in soft tissue sarcomas involving neurovascular structure. AB - BACKGROUND AND PURPOSE: To evaluate intraoperative brachytherapy in the management of soft tissue sarcomas involving neurovascular structures, its impact on local control and complications. PATIENTS AND METHODS: Between 01/1989 and 12/2002, 98 patients received an intraoperative implant in conjunction with conservative surgery. Brachytherapy was part of the initial treatment (79 cases) or performed in recurrent disease (19 cases). We studied primary sarcomas involving neurovascular structures treated with conservative surgery and intraoperative brachytherapy (n = 6) or intraoperative brachytherapy and external irradiation (n = 73). Conservative surgery was performed as first treatment (51 cases), after chemotherapy (21 cases) and after primary external radiation (seven cases). Brachytherapy was performed according to Paris system rules. Patients were loaded with Iridium 192 (64 cases) or connected to a Microselectron PDR (15 cases). Mean dose given by brachytherapy was 20 Gy. Mean dose given of external radiotherapy was 46 Gy. RESULTS: With a median follow-up of 58 months, 5-year actuarial survival was 69% and local free disease at 5 years was 90%. Acute side effects occurred in 22/79 requiring surgical repair in 10 patients. Late side effects occurred in 35/79. No patient required amputation for complications. Prognostic factors were studied for the occurrence of acute and late side-effects and local control. CONCLUSIONS: Intraoperative brachytherapy is efficient with excellent local control rates in soft tissue sarcomas presenting with neurovascular involvement and offers an acceptable conservative option. PMID- 16376448 TI - Protease (PrA and PrB) and prolyl and arginyl aminopeptidase activities from Debaryomyces hansenii as a function of growth phase and nutrient sources. AB - The effects of nutrient sources and growth phase of Debaryomyces hansenii on the protease (PrA and PrB) and aminopeptidase (prolyl-[PAP] and arginyl-[AAP] aminopeptidases) activities were investigated. These activities were also monitored during growth on a whole sarcoplasmic muscle protein extract (WSPE) and on an equivalent medium but free of compounds under 10 kDa (SPE>10 kDa). The levels of specific protease and aminopeptidase activities were higher when cells were grown in urea and dipeptides than when grown in either ammonium or free amino acids as nitrogen sources. The level of each aminopeptidase (PAP or AAP) activity was preferentially induced by its own substrate (ProLeu or LysAla), suggesting a role in the utilization of exogenous peptides. Higher specific activities for all proteolytic enzymes were detected when using acetate as carbon source. The time course experiments carried out on urea or sarcoplasmic protein containing media revealed an increase in all activities during transition and advanced stages of stationary phase of growth. In muscle protein extracts, the absence of low molecular mass nutrients (SPE>10 kDa) initially induced the production of PrA, PrB, and AAP activities, possibly involved in the breakdown of muscle oligopeptides. PMID- 16376449 TI - Lactobacillus plantarum inhibits growth of Listeria monocytogenes in an in vitro continuous flow gut model, but promotes invasion of L. monocytogenes in the gut of gnotobiotic rats. AB - The ability of the pediocin AcH producing Lactobacillus plantarum DDEN 11007 and its non-producing plasmid-cured isogenic variant, DDEN 12305 to prevent the persistence and growth of Listeria monocytogenes EP2 in two gastrointestinal (GI) tract models was examined. In vitro studies conducted in a two-stage continuous flow system showed that L. plantarum DDEN 11007 inhibited L. monocytogenes EP2 under these conditions, while less effect was seen of the non-bacteriocin producing variant. The inhibitory effect was more pronounced at pH 5 than at pH 7. No effect on persistence of L. monocytogenes in the GI tract was seen in gnotobiotic rats colonized with either the pediocin AcH producing or the non bacteriocin producing variant of L. plantarum when compared to rats inoculated with L. monocytogenes EP2 alone. Surprisingly, inoculation of the gnotobiotic animals with either of the L. plantarum strains prior to inoculation with L. monocytogenes EP2 resulted in increased occurrence of L. monocytogenes in liver and spleen when compared to the animals inoculated with L. monocytogenes EP2 alone. Our results indicate that the presence of L. plantarum in the gut of gnotobiotes facilitates L. monocytogenes invasion by an unknown mechanism. This observation is however not necessarily specifically related to L. plantarum, and should not be interpreted as the expected effect in animals carrying a conventional intestinal microflora. PMID- 16376450 TI - Mice heterozygous for the Mdr2 gene demonstrate decreased PEMT activity and diminished steatohepatitis on the MCD diet. AB - BACKGROUND/AIMS: The administration of a methionine and choline deficient (MCD) diet to mice serves as an animal model of NASH. The multidrug resistant 2 (Mdr2) P-glycoprotein encodes for the canalicular phospholipid transporter, and Mdr2 (+/ ) mice secrete 40% less phosphatidylcholine than wild-type mice. We have hypothesized that phosphatidylethanolamine-N-methyl transferase (PEMT) up regulation is a consequence of MCD diet administration, and is important for the pathogenesis of steatohepatitis in this model. However, the effect of decreased phosphatidylcholine secretion and modulation of PEMT on the development of diet induced steatohepatitis in Mdr2 (+/-) mice has not been explored. Thus, the purpose of the study is to examine the effects of the MCD diet on Mdr2 (+/-) mice. METHODS: Mdr2 (+/-) and Mdr2 (+/+) mice were treated with an MCD or control diet for up to 30 days, and the severity of steatohepatitis, PEMT activity and hepatic S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH) levels were measured. RESULTS: Serum ALT levels, hepatic inflammation, and PEMT activity were significantly lower, and hepatic SAM:SAH ratios were significantly higher in Mdr2 (+/-) mice at 7 and 30 days on the MCD diet. CONCLUSIONS: Mdr2 (+/-) mice have diminished susceptibility to MCD diet-induced NASH, which is associated with a relative decrease in PEMT activity and increased SAM:SAH ratios. PMID- 16376451 TI - Filter design for cancellation of baseline-fluctuation in needle EMG recordings. AB - Appropriate cancellation of the baseline fluctuation (BLF) is an important issue when recording EMG signals as it may degrade signal quality and distort qualitative and quantitative analysis. We present a novel filter-design approach for automatic cancellation of the BLF based on several signal processing techniques used sequentially. The methodology is to estimate the spectral content of the BLF, and then to use this estimation to design a high-pass FIR filter that cancel the BLF present in the signal. Two merit figures are devised for measuring the degree of BLF present in an EMG record. These figures are used to compare our method with the conventional approach, which naively considers the baseline course to be of constant (without any fluctuation) potential shift. Applications of the technique on real and simulated EMG signals show the superior performance of our approach in terms of both visual inspection and the merit figures. PMID- 16376452 TI - Fast parametric imaging algorithm for dual-input biomedical system parameter estimation. AB - Medical parametric imaging with dynamic positron emission tomography (PET) plays an increasingly potential role in modern biomedical research and clinical diagnosis. The key issue in parametric imaging is to estimate parameters based on sampled data at the pixel-by-pixel level from certain dynamic processes described by valid mathematical models. Classic nonlinear least squares (NLS) algorithm requires a "good" initial guess and the computational time-complexity is high, which is impractical for image-wide parameter estimation. Although a variety of fast parametric imaging techniques have been developed, most of them focus on single input systems, which do not provide an optimal solution for dual-input biomedical system parameter estimation, which is the case of liver metabolism. In this study, a dual-input-generalized linear least squares (D-I-GLLS) algorithm was proposed to identify the model parameters including the parameter in the dual input function. Monte Carlo simulation was conducted to examine this novel fast algorithm. The results of the quantitative analysis suggested that the proposed technique could provide comparable reliability of the parameter estimation with NLS fitting and accurately identify the parameter in the dual-input function. This method may be potentially applicable to other dual-input biomedical system parameter estimation as well. PMID- 16376453 TI - COX-2 inhibitors suppress lung cancer cell growth by inducing p21 via COX-2 independent signals. AB - COX-2 has been implicated in the control of human non-small cell lung carcinoma (NSCLC) cell growth. The mechanisms by which COX-2 exerts its mitogenic effects have not been entirely elucidated, but stimulation of prostaglandin E2 production and alterations in the expression of the cyclin-dependent kinase inhibitor p21(WAF-1/CIP1/MDA-6)(p2i) have been suggested. Here, we demonstrate that two COX 2 inhibitors (NS398 and Nimesulide) inhibit proliferation and induce apoptosis in NSCLC cells, and these effects were associated with induction of p21 mRNA and protein expression. However, the anti-growth effect of the COX-2 inhibitors and their ability to induce p21 were not affected by COX-2 siRNA suggesting that their actions were COX-2 independent. Instead, activation of the MEK-1/Erk pathway was necessary since COX-2 inhibitors stimulated the phosphorylation of ERKs, and their effects were blocked by PD98095, an inhibitor of this pathway. Furthermore, we show that both NS398 and Nimesulide induced p21 gene promoter activity and this was prevented by PD98095. COX-2 inhibitors increased nuclear protein binding to the Spl site in the promoter region of the p21 gene. Consistent with a role for p21, we found that p21 antisense oligonucleotides prevented the effects of COX-2 inhibitors on cell growth. In summary, our results suggest that COX-2 inhibitors suppress NSCLC cell growth by inducing the expression of the p21 gene through MEK-1/ERK signaling and DNA-protein interactions involving Spl. These observations unveil a mechanism for p21 gene regulation by COX-2 inhibitors in lung carcinoma cell growth and this pathway represents a potential target for therapy. PMID- 16376454 TI - The possible involvement of peroxidase in defense of yellow lupine embryo axes against Fusarium oxysporum. AB - Peroxidase activity (EC 1.11.1.7) towards phenolic substrates, i.e. pyrogallol, syringaldazine and guaiacol, and ascorbate peroxidase activity (EC 1.11.1.11) were analyzed in embryo axes of Lupinus luteus L. cv. Polo cultured on Heller medium for 96h after inoculation with the necrotrophic fungus Fusarium oxysporum f.sp. Schlecht lupini. Four variants were compared: inoculated embryo axes cultured with 60mM sucrose (+Si) or without it (-Si), and non-inoculated embryo axes cultured with 60mM sucrose (+Sn) or without it (-Sn). Between 0 and 96h of culture, peroxidase activity towards the phenolic substrates increased in all variants except -Si, where a decrease was noted in peroxidase activity towards syringaldazine and guaiacol, but not towards pyrogallol. In +Si tissues, a considerable increase in enzyme activity towards these substrates was recorded starting from 72h of culture. Lignin content of +Si tissues increased already at the first stage of infection, i.e. 24h after inoculation. Additionally, in +Sn tissues, high ascorbate peroxidase activity was observed during the culture. Its activity increased in +Si tissues, beginning at 72h after inoculation. However, this was lower than in +Sn tissues. At 72h after inoculation, a considerably stronger development of the infection was observed in -Si than in +Si tissues during our earlier research [Morkunas, I. et al., 2005. Sucrose-stimulated accumulation of isoflavonoids as a defense response of lupine to Fusarium oxysporum. Plant Physiol Biochem 2005; 43: 363-73]. Both peroxidases assayed towards phenolic substrates and ascorbate peroxidase was less active in -Si tissues than in -Sn tissues. Hydrogen peroxide concentration was much higher in Si than in +Si tissues. These results indicate that peroxidases may be some of the elements of the defense system that are stimulated by sucrose in yellow lupine embryo axes in response to infection caused by F. oxysporum. PMID- 16376455 TI - Characterization of alternative oxidase (AOX) gene expression in response to methyl salicylate and methyl jasmonate pre-treatment and low temperature in tomatoes. AB - Methyl salicylate (MeSA) vapor increased resistance against chilling injury (CI) in freshly harvested pink tomatoes. The expression patterns of alternative oxidase (AOX) before and during the chilling period demonstrated that pre treatment of tomato fruit with MeSA vapor increased the transcript levels of AOX. We used 4 EST tomato clones of AOX from the public database that belong to two distinctly related families, 1 and 2 defined in plants. Three clones were designated as LeAOX1a, 1b and 1c and the fourth clone as LeAOX2. Using RT-PCR, 1a and 1b genes were found to be expressed in leaf, root and fruit tissues, but 1c was expressed preferentially in roots. RNA transcript from LeAOX1a of AOX subfamily 1 was present in much greater abundance than 1b or 1c. The presence of longer AOX transcripts detected by RNA gel blot analysis in cold-stored tomato fruit was confirmed to be the un-spliced pre-mRNA transcripts of LeAOX1a and LeAOX1b genes. Intron splicing of LeAOX1c gene was also affected by cold storage when it was detected in roots. This alternative splicing event in AOX pre-mRNAs molecules occurred, preferentially at low temperature, regardless of mRNA abundance. Transcript levels of several key genes involved in RNA processing (splicing factors: 9G8-SR and SF2-SR, fibrillarin and DEAD box RNA helicase) were also affected by changes in storage temperature. The aberrant splicing event in AOX pre-mRNA and its possible association with the change in expression of genes involved in RNA processing in tomato fruit having chilling disorder was discussed. PMID- 16376456 TI - Comparison of floor sanitation methods. AB - Two methods for cleaning waxed polyvinylchloride and porcelain gres hospital room floors were compared in order to determine their decontamination capacity: dry wiping followed by damp washing, and damp washing followed by dry wiping. Dry wiping followed by damp washing did not produce any significant reduction in the average bacterial load. However, damp washing followed by dry wiping reduced the bacterial load for both types of flooring. The difference was statistically significant. PMID- 16376457 TI - Multicentre study on hand hygiene facilities and practice in the Mediterranean area: results from the NosoMed Network. AB - Hand hygiene literature is scarce in the southern Mediterranean area. In order to establish a baseline position, a study was performed in four Mediterranean countries. Seventy-seven hospital wards in 22 hospitals were enrolled and information on hand hygiene practice and facilities were collected. The overall compliance rate was very low (27.6%), and was significantly higher where the perceived risk was considered to be high. Intensive care units showed the highest level of compliance. Analysis by country indicated higher compliance in Egypt (52.8%) and Tunisia (32.3%) compared with Algeria (18.6%) and Morocco (16.9%). Facilities for hand hygiene, particularly consumables, were shown to be deficient. Multi-approach programmes combining the production of official local recommendations, education and regular evaluation of hand hygiene practice are much needed to improve the present situation. PMID- 16376458 TI - Molecular cloning and characterization of FGLamide allatostatin gene from the prawn, Macrobrachium rosenbergii. AB - Allatostatins are important regulatory neuropeptides that inhibit juvenile hormone (JH) biosynthesis by the corpora allata (CA) in insects. However, to date, the structure and expression of the gene encoding allatostatins have not been reported in any species other than insects. In this study, we used a combination of a semi-nested polymerase chain reaction (PCR) and screening of a central nervous system cDNA library of Macrobrachium rosenbergii to isolate and sequence a cDNA clone (2885 bp) encoding a 701 amino acid FGLamide allatostatin precursor polypeptide. This is the first reported allatostatin gene in crustacean. The deduced precursor was conceptually split into at least 35 FGLamide allatostatins at dibasic cleavage sites (Lys and Lys/Arg), far more than reported for any other known FGLamide allatostatin precursors from insects (13-14 allatostatins). Reverse transcription-polymerase chain reaction (RT-PCR) analysis demonstrated that the gene was expressed in the brain, gut, thoracic and abdominal ganglia, but not in the heart, muscle, ovary, gill, or hepatopancreas. Furthermore, developmentally-dependent expression of the gene was observed in the brain and thoracic ganglia of the prawn by using semi-quantitative RT-PCR analysis. PMID- 16376459 TI - Identification of adipocyte differentiation-related regulatory element for adrenomedullin gene repression (ADRE-AR) in 3T3-L1 cells. AB - Adrenomedullin (AM), a potent vasodilator peptide, has been suggested to act against cardiovascular complications and insulin resistance in the metabolic syndrome. We have already reported the AM gene repression in the early phase of adipocyte differentiation of NIH 3T3-L1 cells. Here we show adipocyte differentiation-related regulatory element for AM gene repression (ADRE-AR) in 36 bp region (-2135/-2100) of the AM gene. 3T3-L1 cells were induced to differentiate to adipocytes by insulin, dexamethasone and 3-isobutyl-1 methylxanthine. On the third day of differentiation, the promoter function was analyzed using the reporter plasmids, which contain the promoter region of AM gene (-4616/+108) in pGL3-basic luciferase reporter vector. The promoter activity decreased to about 20% in 3T3-L1 adipocytes when compared with 3T3-L1 preadipocytes, and a 36-bp region (-2135 to -2100) upstream from the transcription initiation site of the AM gene was necessary for higher AM gene expression in preadipocytes. This 36-bp ADRE-AR contains three copies of G/AAAA sequence (5'-GAAATGAAAGTAAAA-3') (-2124/-2110), which are conserved between mouse and human, and the introduction of mutations in each copy of G/AAAA sequence decreased the promoter activity in preadipocytes and adipocytes. Electrophoretic mobility shift assay showed that the full-length ADRE-AR was specifically bound by a certain nuclear protein(s). The present study has raised the possibility that ADRE-AR may play important roles in the AM gene expression in preadipocytes, and that the AM gene may be repressed through the ADRE-AR in adipocytes. PMID- 16376460 TI - [Aortic involvement in giant cell arteritis. A prospective follow-up of 11 patients using computed tomography]. AB - PURPOSE: To determine clinical and radiological features, using computed tomography (CT-scan) in patients with aortic involvement related to giant cell arteritis (GCA), and to assess both clinical and CT-scan outcome after therapy institution. METHODS: Aortic involvement due to GCA was investigated in all patients, using CT-scan at diagnosis, and at 3, 6 and 12 months follow-up after therapy institution. RESULTS: The 11 consecutive patients consisted of 4 men and 7 women with mean age of 64.5 years. Patients exhibited: constitutional symptoms (N=9; 82%), dorsalgia (N=3; 27%), clinical signs of GCA (N=3; 27%) and of upper limb large vessel impairment (N=6; 55%). CT-scan showed aortitis involving both thoracic and abdominal aorta (N=6; 55%), abdominal (N=2; 18%) or thoracic aorta (N=2; 18%) and thoracic aortic aneurysm (N=1; 9%). At one-year follow-up, CT-scan revealed: complete resolution (N=7; 64%) and improvement (N=3; 27%) of aortic damage; the patient, who had thoracic aortic aneurysm, underwent surgical treatment, as aortic lesion remained unchanged on CT-scan. CONCLUSION: Our study underlines that CT-scan is a helpful test in diagnosis and follow-up of aortic involvement in patients with GCA. PMID- 16376461 TI - [Factors associated with hyperhomocysteinemia in inflammatory bowel disease: prospective study in 81 patients]. AB - BACKGROUND: A high prevalence (52%) of hyperhomocysteinemia is observed in Crohn disease (CD), however it is not well documented in ulcerative colitis (UC). Furthermore, in the different works studying hyperhomocysteinemia the associated factors are different. AIM: Prospective evaluation of hyperhomocysteinemia in inflammatory bowel disease (IBD) patients, of the risk factors and the determination of a potential risk of colorectal carcinoma in case of hyperhomocysteinemia. PATIENTS AND METHODS: IBD patients followed in our department were prospectively recruited between November 2003-September 2004. To be included patients should have passed a coloscopy in the two years. Patients with kidney failure or drugs supposed, to interfere with homocysteine metabolism (folates, vitamin B12, methotrexate) were excluded from the study. The following parameters were analysed: age, sex, clinical activity indexes (CDAI for Crohn disease and CAI for ulcerative colitis), length-extent and type of the disease (CD or UC), smoking, plasma homocystein concentration, folates and vitamin B12. RESULTS: Eighty-one patients (60 CD, 21 UC, mean age 43.8 +/- 17.3) were included, 30 had an active disease at inclusion and 16 were smokers. The prevalence of high homocystein concentration was 55.6%. In univariate analysis a low rate of folates was the only risk factor for a high homocystein concentration (74 vs. 52.8%; P = 0.018). Smoking was almost an associated factor. In multivariate analysis, a low rate of folate was the only risk factor of hyperhomocysteinemia, OR = 3.59 [1.27-10.17]. Five endoscopic lesions considered as precancerous were described; these patients had all a hyperhomocysteinemia. CONCLUSION: The prevalence of hyperhomocysteinemia is high in UC and in CD. A low folate rate is the only risk factor observed in our study. There is a possible link between colorectal cancer and hyperhomocysteinemia. A high Plasma homocystein concentration must be search in inflammatory bowel disease patients and a substitutive treatment of folates and vitamin B12 is necessary in case of hyperhomocysteinemia. PMID- 16376462 TI - The key role of micronutrients. AB - Micronutrients play a central role in metabolism and in the maintenance of tissue function, but effects in preventing or treating disease which is not due to micronutrient deficiency cannot be expected from increasing the intake. There is a highly integrated system to control the flux of micronutrients in illness, and this demonstrates just how important the body perceives the micronutrients to be. An adequate intake therefore is necessary to sustain metabolism and tissue function, but provision of excess supplements to individuals who do not need them may be harmful. Clinical benefit is most likely in those individuals who are severely depleted and at risk of complications, and is unlikely if this is not the case. Much more research is needed to characterise better markers of micronutrient status both in terms of metabolic effects and antioxidant effects, so that at-risk patients can be identified and supplementation modified accordingly. Large-scale trials of different doses of micronutrients are required with precise outcome markers to optimise intakes in different groups of patients as well as in the general population. PMID- 16376463 TI - Estimates of body height in adult inpatients. AB - BACKGROUND AND AIMS: Different estimates of body height are broad applied in clinical practice, although its accuracy and precision are few known. In order to clarify the applicability of these estimates, concerned with the hypothesis that it is of too few accuracy and precision, the authors compared patient's actual height to their self reported height (SRH), to the value estimated by the equation proposed by World Health Organization (WHO-EQ) and total arm span (TAS). METHODS: 334 adult inpatients had their measured height (gold standard) compared to each estimate through paired t-test. Proportion of patients who had unacceptable errors (difference > 5 cm) is described. RESULTS: Mean difference between actual height and SRH was 1.9 cm, 3.3 cm for WHO-EQ and 5.7 cm for TAS. However, even accepting a 5 cm difference in relation to actual height, the percentage of patients who had errors bigger than so was 16.9% for SRH, 41.9% for WHO-EQ and 51.2% for TAS. A new equation could be obtained from this sample, but, despite methodologically adequate, it wrongly predicted height in more than 5 cm in 27.8% of patients. CONCLUSIONS: Tested estimates appeared to be little reliable to be applied to patients confined to bed due to lack of precision and accuracy. PMID- 16376464 TI - Intraduodenal infusion of alpha-ketoglutarate decreases whole body energy expenditure in growing pigs. AB - BACKGROUND AND AIMS: alpha-Ketoglutarate (AKG) has been suggested to play a particular role as an oxidative fuel for the gut, and thus may have a sparing function for fuels such as glutamate and aspartate. Using the pig model we aimed to quantify how the route of administration (intravenous, i.v.; intragastric, i.g.; intraduodenal, i.d.) affects AKG utilization, whole body energy expenditure (EE) and nutrient oxidation. METHODS: Pigs (15 kg) were supplied with a complete nutrient solution (NS) via catheters. To explore the metabolic effects of AKG, 1.0 g AKG kgBW(-1)d(-1) was infused simultaneously with the NS using either the i.d., i.v. or i.g. route. [1-(13)C]AKG (15 mg kgBW(-1)) was infused i.d., i.v. or i.g., respectively, for 3h. AKG utilization (AKG UTIL) was estimated as AKG UTIL=100-(13)C recovery (% of (13)C dose). (13)C recovery was calculated from the (13)C enrichment in breath CO(2) and the whole-body CO(2) production. RESULTS: AKG infusion and NS via the i.d. route resulted in a reduced AKG UTIL (40.1+/ 6.7) as compared to the i.v. route (62.9+/-2.4, P<0.001) and i.g. route (62.3+/ 1.6, P<0.001). The total EE was lower with the i.d. route of AKG and NS (745+/-68 kJkgBW(-0.62)d(-1)) as compared to the i.v. route (965+/-54 kJkgBW(-0.62)d(-1), P<0.005) and i.g. route (918+/-43 kJkgBW(-0.62)d(-1), P<0.005). Carbohydrate oxidation was increased with the i.d. route (38.2g+/-3.4 kgBW(-0.62)d(-1)) as compared to the i.v. route (27.8+/-2.9 gkgBW(-0.62)d(-1), P<0.08) and i.g. route (23.9+/-8.5 gkgBW(-0.62)d(-1), P<0.05). Fat oxidation was decreased (2.1+/-1.9 gkgBW(-0.62)d(-1); P<0.001) with the i.d. route as compared to the i.v. route (11.5+/-1.4 gkgBW(-0.62)d(-1), P<0.001) and i.g. route (11.9+/-3.1 gkgBW(-0.62)d( 1), P<0.001). CONCLUSIONS: The i.d. infusion of AKG in combination with the NS affected the whole body EE and nutrient oxidation, in comparison to that obtained with the i.v. and i.g. routes. It was concluded that the i.d. administration of AKG markedly controlled the nutrient partitioning in the oxidation processes. Finally, in contrary to the observations with glutamine or glutamate, a considerable percentage of the AKG infusion was retained in the body irrespective of the route of administration. PMID- 16376465 TI - Human tumor membrane vesicles modified to express glycolipid-anchored IL-12 by protein transfer induce T cell proliferation in vitro: a potential approach for local delivery of cytokines during vaccination. AB - Administration of soluble human interleukin-12 (hIL-12) has been shown to induce a potent anti-tumor response. However, the use of soluble hIL-12 is hindered by its cytotoxicity when systemically administered and the difficulty of transferring multiple genes into primary tumor cells. In this study, we developed a membrane-anchored hIL-12 and expressed it on tumor membrane vesicles to deliver and confine IL-12 to the vaccination site. We constructed a glycolipid-anchored hIL-12 (GPI-hIL-12) by fusing the coding region of p35 and p40 subunits of hIL-12 with the GPI-signal sequence of CD59 at the C-terminal ends. The two subunits were processed correctly and expressed as a GPI-anchored disulfide-linked heterodimeric protein on the cell surface. GPI-hIL-12 cells induced proliferation of activated T cells and augmentation of allogeneic T cell generation in an MLR assay. Purified GPI-hIL-12 was efficiently intercalated onto isolated tumor cell membrane vesicles prepared from various human tumor cell lines. Further, the incorporation of GPI-hIL-12 onto tumor membrane vesicles induced proliferation of T cells and the release of IFN-gamma by activated T cells. Notably, GPI-hIL-12 enhanced the proliferative response initiated by CD80, a principal costimulatory molecule for T cell activation. These studies suggest that tumor membrane vesicles modified with GPI-anchored cytokines can be used to create potent immunogenic tumor vaccines for use in human immunotherapy. Since protein transfer can be used to modify tumor membrane vesicles obtained from surgical specimens, this approach offers a useful alternative to gene therapy as a means of developing cancer vaccines. PMID- 16376467 TI - Gender differences in pattern of hip and lumbopelvic rotation in people with low back pain. AB - BACKGROUND: Findings from previous studies suggest gender may affect the pattern of hip and lumbopelvic motion during a multi-segmental movement. To date, no studies have examined movement patterns and low back pain symptom behavior during hip lateral rotation. METHODS: Forty-six people (27 males and 19 females) with low back pain were examined. Three-dimensional kinematic data and low back pain symptoms were recorded during active hip lateral rotation. Percent of maximum lumbopelvic rotation was calculated for each 10% increment of maximum active hip lateral rotation. FINDINGS: Men exhibited a greater percent of maximum lumbopelvic rotation (mean 49.3, SD 13.3) during the first 60% of hip lateral rotation than women (mean 36.2, SD 16.4) (P < 0.01). Nineteen (70.4%) of the men and seven (36.8%) of the women had pain with the hip lateral rotation test (P = 0.02). INTERPRETATION: Men exhibited more lumbopelvic rotation in the early part of hip lateral rotation than women, and hip lateral rotation was more likely to be associated with symptoms in men than women. Greater lumbopelvic motion, earlier in hip lateral rotation, may make men more vulnerable to low back pain associated with hip lateral rotation. Factors that contribute to these gender differences should be investigated further. PMID- 16376468 TI - Distribution of sorbed phenanthrene and pyrene in different humic fractions of soils and importance of humin. AB - Contributions of fulvic-humic acids (FA/HA) and humin (HM) to sorption of phenanthrene (PHE) and pyrene (PYR) in a soil were differentiated using a humic separation procedure after multi-concentration sorption experiments. It was found that the amount of solutes in FA/HA did not change significantly after 48 h, while that in HM increased continuously and slowly up to the end of the experimental period (720 h), indicating that HM was the main region for slow sorption. Based on the fitting results using Freundlich equation, it was found that nonlinearity of both solutes was greater in HM than in FA/HA, consistent with the sorption characteristics of individually extracted HA and HM in a separate experiment. The observed nonlinearity of the solute distribution was confirmed by using three other soil samples with organic carbon contents ranging from 0.7 to 7.9%. Distribution dynamics of PHE and PYR among various fractions were also discussed. PMID- 16376469 TI - Cadmium and lead concentrations in Skrjabinotaenia lobata (Cestoda: Catenotaeniidae) and in its host, Apodemus sylvaticus (Rodentia: Muridae) in the urban dumping site of Garraf (Spain). AB - The present study evaluates the parasitological model constituted by the wood mouse (Apodemus sylvaticus) and its intestinal cestode (Skrjabinotaenia lobata) as a potential bioindicator of Cd and Pb in the urban dumping site of Garraf near the city of Barcelona (Spain) and in Begues (reference site). Tissues and respective S. lobata specimens of 38 wood mice captured in Garraf and Begues were analyzed for Cd and Pb by means of ICP-MS. Higher cadmium levels in S. lobata were found only in respect to the muscular levels of their hosts. Nevertheless, lead levels were 8.5-, 53.2- and 81.4-fold higher in S. lobata than kidney, liver and muscle levels of A. sylvaticus from Garraf, respectively. Thus, the proposed model seems to be a promising bioindicator to evaluate environmental lead exposure in terrestrial habitats. In addition, all available data on lead bioaccumulation by cestode parasites of terrestrial mammals are generally discussed. PMID- 16376470 TI - The role of evidence in alternative medicine: contrasting biomedical and anthropological approaches. AB - The growth of alternative medicine and its insurgence into the realms of the biomedical system raises a number of questions about the nature of evidence. Calls for 'gold standard' randomised controlled trial evidence, by both biomedical and political establishments, to legitimise the integration of alternative medicine into healthcare systems, can be interpreted as deeply political. In this paper, the supposed objectivity of scientific, biomedical forms of evidence is questioned through an illumination of the multiple rhetorics embedded in the evidence-based medicine phenomenon, both within biomedicine itself and in calls for its use to evaluate alternative therapeutic systems. Anthropological notions of evidence are constructed very differently from those of biomedical science, and offer a closer resonance with the philosophy of alternative medicine. Examples are given of the kinds of evidence produced by anthropologists researching alternative medicine. Ethnographic evidence of 'what works' in alternative medicine includes concepts such as transcendent, transformational experiences; changing lived-body experience; and the gaining of meaning. It is proposed that the promotion of differently constructed modes of evidence can be used to legitimise alternative medicine by widening the definition of what works in therapy, and offering a critique of what people feel is lacking from much of orthodox medical care. PMID- 16376472 TI - The political contexts of evidence-based medicine: policymaking for daily hemodialysis. AB - Policymakers and clinicians increasingly rely on evidence-based medicine (EBM) to make decisions about insurance coverage and clinical treatment. Conflicting value judgments about evidence and pressures exerted by stakeholders render health policymaking a political process. This paper examines how value judgments become embedded in the process of improving medical outcomes by focusing on health policymaking. Specifically, this paper highlights how EBM is variably used as a standard for decision-making depending on perceived risks by policymakers and what is on the competing agenda. I draw upon the case study of the policymaking process for the recent US bill, H.R. 1004: Kidney Patient Daily Dialysis Act, which would legislate daily hemodialysis (DHD) as a new renal replacement therapy modality, and provide federal medicare funding of hemodialysis from 3 to 6 times per week. DHD constitutes an ideal case study with which to explore the political underpinnings of EBM. The interpretations of substantial outcome data showing medical, quality of life, and hypothetical economic improvements of DHD over conventional dialysis are currently being contested in the medical and political spheres. Accordingly, the drive for what some stakeholders view as better evidence through randomized clinical trials is central to the debate and policymaking process. This paper underscores how the demand for, the interpretations, the funding for, and the use of evidence render EBM a political endeavor with vital ethical implications for clinical care. PMID- 16376471 TI - The social and cultural shaping of medical evidence: case studies from pharmaceutical research and obstetric science. AB - Most critiques of evidence-based medicine (EBM) focus on the scientific shortcomings of the technique. Social scientists are more likely to criticize EBM for it ideological biases, a criticism that makes sociological sense but is difficult to substantiate. Using data from our studies of (1) the influence of pharmaceutical companies on the conduct and reporting of clinical trials, and (2) obstetric science in the Netherlands (where nearly one-third of births occur at home) we show how the evidence of evidence-based medicine is shaped by forces both structural and cultural. The threats to objective evidence are many, and, if EBM is to be true to its own principles, it must take these threats into account. PMID- 16376473 TI - Antipsychotic-associated neuronal changes in the brain: toxic, therapeutic, or irrelevant to the long-term outcome of schizophrenia? AB - The increasingly wide-spread use of antipsychotics in both adults and children calls for a detailed examination of antipsychotic-associated neuronal changes in the brain, and whether these changes are toxic, therapeutic, or perhaps irrelevant to the outcome of major psychiatric disorders, especially schizophrenia. In this review we will examine the extensive evidence demonstrating both acute and longer-term antipsychotic-associated neurotoxicity and neuroplasticity, as well as the more specific cellular changes that appear to underlie these phenomena. These include changes in proteins affecting cell survival, impairment of the mitochondrial respiratory chain, increases in DNA fragmentation, injury to dendritic microtubules, increases in dopamine-generated reactive oxygen species, changes in cell morphology, and rapid induction of apoptosis. We shall also examine the correlation between these changes and alterations in gross brain structure. There appears to be a disjunction between the widespread cellular and gross structural brain changes in schizophrenia, and the duration of illness, expression of symptoms, and response to treatment. We shall explore possible explanations for this apparent paradox. PMID- 16376474 TI - Insights into metal ion binding in phospholipases A2: ultra high-resolution crystal structures of an acidic phospholipase A2 in the Ca2+ free and bound states. AB - The electrophile Ca(2+) is an essential multifunctional co-factor in the phospholipase A(2) mediated hydrolysis of phospholipids. Crystal structures of an acidic phospholipase A(2) from the venom of Bothrops jararacussu have been determined both in the Ca(2+) free and bound states at 0.97 and 1.60 A resolutions, respectively. In the Ca(2+) bound state, the Ca(2+) ion is penta coordinated by a distorted pyramidal cage of oxygen and nitrogen atoms that is significantly different to that observed in structures of other Group I/II phospholipases A(2). In the absence of Ca(2+), a water molecule occupies the position of the Ca(2+) ion and the side chain of Asp49 and the calcium-binding loop adopts a different conformation. PMID- 16376475 TI - Escherichia coli RecA promotes strand invasion with cisplatin-damaged DNA. AB - The antitumor drug cisplatin causes intrastrand cross-linking of adjacent guanine residues that severely distorts the DNA backbone. These DNA adducts impede the progress of the replisome and may result in replication fork arrest. In Escherichia coli, the response to cisplatin involves the action of the prototypic recombinase RecA. Here we show that RecA can utilize, albeit at reduced levels, oligonucleotides that bear site-specific cisplatin-induced 1,2 d(GpG) intrastrand cross-links in strand invasion reactions. Binding of RecA to cisplatin-damaged oligonucleotides was not affected, indicating that the impediment was in the pairing step. The cognate E. coli single-strand DNA-binding protein specifically stimulated strand invasion particularly with cisplatin-damaged DNA. These results indicate that RecA is capable of processing the major cisplatin-induced lesion via a recombination mechanism. PMID- 16376476 TI - Biochemical analysis of alginate biosynthesis protein AlgX from Pseudomonas aeruginosa: purification of an AlgX-MucD (AlgY) protein complex. AB - AlgX was found to be an essential protein for alginate biosynthesis, but its function is unknown. In this study, an isogenic, marker-free algX-knock out mutant was generated. In-frame fusions of algX with phoA and lacZ were analysed, respectively. No LacZ-activity was detected, but the PhoA fusion showed alkaline phosphatase activity. These data indicated that the C-terminus of AlgX is located in the periplasm, but is not required for protein function. Accordingly, AlgX with C-terminal fusion of strep tag II restored alginate production in the algX negative mutant and was purified under native conditions from periplasmic and crude cell extracts, respectively. AlgX was identified by MALDI/TOF-MS analysis of tryptic peptides. TritonX-100 mediated solubilisation of cytoplasmic membrane and subsequent strep tag II affinity chromatography led to purification of an AlgX-MucD (AlgY) protein complex as identified by MALDI/TOF-MS analysis. This data suggested a protein-protein interaction between AlgX and MucD (AlgY) with a 1:1 stoichiometry. Thus AlgX might exert its function via interaction with MucD (AlgY). Immunoelectron microscopic localisation of AlgX-strep tag II suggested a localisation close to the cytoplasmic membrane. PMID- 16376477 TI - New insights on brain stem death: from bedside to bench. AB - As much as brain stem death is currently the clinical definition of death in many countries and is a phenomenon of paramount medical importance, there is a dearth of information on its mechanistic underpinnings. A majority of the clinical studies are concerned only with methods to determine brain stem death. Whereas a vast amount of information is available on the cellular and molecular mechanisms of cell death, rarely are these studies directed specifically towards the understanding of brain stem death. This review presents a framework for translational research on brain stem death that is based on systematically coordinated clinical and laboratory efforts that center on this phenomenon. It begins with the identification of a novel clinical marker from patients that is related specifically to brain stem death. After realizing that this "life-and death" signal is related to the functional integrity of the brain stem, its origin is traced to the rostral ventrolateral medulla (RVLM). Subsequent laboratory studies on this neural substrate in animal models of brain stem death provide credence to the notion that both "pro-life" and "pro-death" programs are at work during the progression towards death. Those programs (mitochondrial functions, nitric oxide, peroxynitrite, superoxide anion, coenzyme Q10, heat shock proteins and ubiquitin-proteasome system) hitherto identified from the RVLM are presented, along with their cellular and molecular mechanisms. It is proposed that outcome of the interplay between the "pro-life" and "pro-death" programs (dying) in this neural substrate determines the final fate of the individual (being dead). Thus, identification of additional programs in the RVLM and delineation of their regulatory mechanisms should shed new lights on future directions for clinical management of life-and-death. PMID- 16376478 TI - Primary lymphoma of the cervix: unusual location for a common disease. PMID- 16376479 TI - Prenatal diagnosis of beta-thalassemia in Southern China. AB - OBJECTIVE: To control the birth of thalassemic children in Southern China. STUDY DESIGN: DNA-based diagnosis was offered on fetal tissues in pregnancies when beta globin gene mutations were identifiable in both parents using polymerase chain reaction (PCR)-reverse dot blot (RDB) assay. An automated high-performance liquid chromatography (HPLC) system was used to analyze fetal hemoglobin in pregnancies when mutation was unidentified in at least one parent. Fetal samplings were collected by chorionic villi sampling (CVS) in the first trimester, and by amniocentesis or cordocentesis in the second trimester. Maternal contamination of fetal DNA was ruled out by short tandem repeats (STR) analysis. RESULTS: Five hundered and forty-five fetuses of 540 at-risk pregnancies were performed prenatal diagnosis. Out of 540 fetuses tested by DNA analysis, 150 were found to be normal, 257 were carriers, whereas 133 were affected. Out of five fetuses diagnosed by HPLC, one fetus was affected and four were unaffected. Totally, 133 pregnancies with affected fetuses, except for one twin pregnancy, were voluntarily terminated, leading to a marked reduction of severe beta-thalassemia in this region. CONCLUSIONS: Our prenatal diagnosis strategy proved to be highly effective. DNA- and HPLC-based testing could enable prenatal diagnosis of beta thalassemia in all at-risk pregnancies. PMID- 16376480 TI - Laparoscopic surgery versus antegrade scrotal sclerotherapy: Retrospective comparison of two different approaches for varicocele treatment. AB - OBJECTIVE: Both laparoscopic surgery and antegrade sclerotherapy are effective treatment options in the management of varicoceles. However, very limited data comparing these two approaches are available in the literature. We present our experience regarding outcome and complications with each treatment modality. PATIENTS AND METHODS: During a 10-year period, 122 cases of laparoscopic varicocelectomy (LV) and 108 cases of antegrade sclerotherapy (AS) were performed in our institution. Diagnosis and postoperative results were established clinically and using Doppler ultrasonography. Data regarding failure rate, complications, operative time and length of hospital stay of each procedure were retrospectively evaluated. RESULTS: Median follow-up was 59 months (5-130). Failure rates for LV and AS were 4.9% and 15.7%, respectively (p < 0.01). Complications occurred in 13.1% after LV, including 13 cases (10.7%) of hydrocele formation, and 4.6% after AS. Hydroceles following LV were significantly more frequent in patients with compared to patients without previous inguinal surgery (27.8% vs. 8.5%; p < 0.05). There was no significant difference between LV and AS regarding operative time (36 vs. 34 min, p > 0.05) and hospital stay (2.2 vs. 2.1 days, p > 0.5). CONCLUSIONS: In our experience, LV was more effective than AS in correcting varicoceles. Complications other than hydroceles were higher after AS. The higher incidence of postoperative hydrocele formation following LV warrants strategies such as the lymphatic sparing approach, especially in patients with previous inguinal surgery. PMID- 16376481 TI - Promoter cloning and characterisation of the transcriptional regulation of the human thyrostimulin A2 subunit. AB - The novel heterodimeric glycoprotein hormone thyrostimulin consists of two unique subunits, A2 and B5. To understand its yet unknown transcriptional regulation, we characterised the 3.1-kb immediate 5'-flanking region of the human A2 gene localised on chromosome 11q13. In transient transfection assays this sequence exhibited promoter activity, which could be confined to nucleotides -506 to -347 relative to the ATG start codon. Interestingly, this minimal promoter appeared to be non-tissue-specific. Deletional, mutational and gel shift analyses revealed regulatory elements that are essential for the regulation of the A2 gene expression. Another noteworthy feature of this gene is the presence of silencer elements upstream and downstream of the promoter. To surmise, our results provide an initial step toward a detailed analysis of the underlying molecular mechanisms of the human thyrostimulin gene expression. PMID- 16376482 TI - Evaluation of danger from fermentation-induced spontaneous ignition of wood chips. AB - Recently we conducted investigations in biological wastes because large pile-up storage of waste wood chips and others caused many fires in Japan. This paper shows the experimental results on wood chips with thermal analysis, by using a Thermogravimetry-differential thermal analysis (TG-DTA) and micro calorimeters, and as well with spontaneous ignition measurements, such as a UN wire mesh cube tester and a spontaneous ignition tester (SIT). Exothermic reaction of wood chips was observed during 45-60 degrees C only by the high sensitive microcalorimeters, TAM and MS 80. This reaction is far apart from the second major reaction by oxidation and is not easy to be recognized by the conventional detectors, like the TG-DTA and the wire mesh cube tester, because their sensitivity cannot meet the strict requirement. Correspondingly, experimental results under the adiabatic condition in the SIT confirmed this theory, in which the onset temperature of spontaneous ignition of wood chips was measured as 50-80 degrees C. This implies that the weak initial reaction at ambient temperatures mainly results from microbial fermentation in the presence of its inherent moisture and possibly gives rise to the further intense combustion sustained by a chemical reaction if the heat cannot be removed from the large scale storage of wood chips. PMID- 16376483 TI - Debridement of burn wounds with a water jet surgical tool. AB - A new instrument, the Versajet system, based on fluid jet technology has recently been advocated as an alternative to standard surgical excisional technique for burn wounds. In this paper we describe our experience with this tool, the technique employed as well as its clinical feasability for debriding partial thickness burn wounds. Seventeen patients were evaluated for clinical efficacy of debridement of their burn wounds using the Versajet system. Burn wound areas of between 0.5 and 5% total body surface (TBSA) involving the face, arm, hand, leg and foot underwent debridement using the Versajet system. The Versajet system was able to sufficiently debride superficial partial thickness and mid-dermal partial thickness wounds for subsequent placement of Biobrane. Deeper partial thickness wounds could be excised for successful autografting. The Versajet system demonstrated some particular advantage in the surgical treatment of superficial to mid-partial thickness burns in areas like the face, hand and foot which can often be difficult to reach and contour with conventional modalities. PMID- 16376484 TI - Mutational analysis of proapoptotic ARTS P-loop domain in common human cancers. AB - Mounting evidence indicates that deregulation of apoptosis is involved in mechanisms of cancer development. ARTS is released from mitochondria into the cytosol during apoptosis, promoting caspase activation by neutralizing the inhibition of inhibitor of apoptotic protein (IAP) on caspases. ARTS contains a P loop GTP-binding domain, which is essential for the apoptotic function of ARTS. The aim of this study was to ascertain whether a genetic alteration of ARTS gene is involved in the development of human cancers possibly by inactivating the apoptotic function of ART. We analyzed the coding region of the P-loop domain of human ARTS gene for the detection of somatic mutations in 100 gastric carcinomas, 100 non-small cell lung cancers, and 69 hepatocellular carcinomas using a polymerase chain reaction (PCR)-based single strand conformation polymorphism (SSCP). However, there was no mutation in the P-loop coding region in the cancers. The data presented here suggest that ARTS P-loop is not frequently mutated in gastric, lung, and hepatocellular carcinomas, and that apoptosis deregulation in cancers is not dependent on the mutation of ARTS gene. PMID- 16376485 TI - P16(INK4a) expression as a potential prognostic marker in cervical pre-neoplastic and neoplastic lesions. AB - An immunohistochemical analysis with monoclonal antibody p16(INK4a) was performed in formalin-fixed, paraffin-embedded samples of 60 cases. The aim was to investigate in biopsies the expression of p16(INK4a) of normal uterine cervical tissue, pre-cancerous and cancerous lesions, and their relation with human papilloma virus (HPV) and HIV status. Three parameters were evaluated: percentage of p16(INK4a) positive cells, reaction intensity, and cell staining pattern. All of these parameters were statistically different when compared among different histological groups. However, logistic regression model showed that the reaction intensity was the best indicator of the expression of p16(INK4a). This expression increases from normal to invasive squamous carcinoma. Sixty-six percent of the patients with CIN grade 1 (CIN1) expressed p16(INK4a) (all these cases were infected with high risk HPV). Our study supports the hypothesis that p16(INK4a) expression in pre-cancerous lesions and cancers can be used to identify HPV transformed cells. Of great interest for routine diagnostic use is the fact that immunohistochemical testing for p16(INK4a) seems to be capable of identifying HPV positive cells and potentially recognizing those lesions with an increased risk of progression to high-grade lesions. PMID- 16376486 TI - Radical radiotherapy for invasive bladder cancer: What dose and fractionation schedule to choose? AB - PURPOSE: To establish the alpha/beta ratio of bladder cancer from different radiotherapy schedules reported in the literature and provide guidelines for the design of new treatment schemes. METHODS AND MATERIALS: Ten external beam radiotherapy (EBRT) and five brachytherapy schedules were selected. The biologically effective dose (BED) of each schedule was calculated. Logistic modeling was used to describe the relationship between 3-year local control (LC3y) and BED. RESULTS: The estimated alpha/beta ratio was 13 Gy (95% confidence interval [CI], 2.5-69 Gy) for EBRT and 24 Gy (95% CI, 1.3-460 Gy) for EBRT and brachytherapy combined. There is evidence for an overall dose-response relationship. After an increase in total dose of 10 Gy, the odds of LC3y increase by a factor of 1.44 (95% CI, 1.23-1.70) for EBRT and 1.47 (95% CI, 1.25-1.72) for the data sets of EBRT and brachytherapy combined. CONCLUSION: With the clinical data currently available, a reliable estimation of the alpha/beta ratio for bladder cancer is not feasible. It seems reasonable to use a conventional alpha/beta ratio of 10-15 Gy. Dose escalation could significantly increase local control. There is no evidence to support short overall treatment times or large fraction sizes in radiotherapy for bladder cancer. PMID- 16376487 TI - Linac-based radiosurgery or hypofractionated stereotactic radiotherapy in the treatment of large cerebral arteriovenous malformations. AB - PURPOSE: We investigate retrospectively clinical outcome after radiosurgery (RS) or hypofractionated stereotactic radiotherapy (HSRT) in patients with large cerebral arteriovenous malformations (AVMs). METHODS AND MATERIALS: This analysis is based on 48 patients with cerebral AVM greater than 4 cm treated with HSRT or RS at our institution. Fifteen patients received HSRT, with 26 Gy median total dose in 4 to 5 fractions, and 33 patients received RS with 17 Gy median total dose in 4 to 5 fractions. Median target volume was 27 cc in HSRT and 7 cc in RS; median maximum diameter was 6 cm and 5 cm, respectively. Seventeen patients experienced intracranial hemorrhage before treatment. Median follow-up was 2.6 years. RESULTS: The 3-year and 4-year actuarial complete obliteration (CO) after HSRT was 17% and 33% and after RS was 47% and 60%, respectively. Actuarial CO was higher in AVMs less than 5 cm (66% vs. 37% after 4 years). Intracranial hemorrhage after HSRT occurred in 3 of 15 patients after 18 months median, and after RS in 7 of 33 patients after 17 months median. Bleeding risk was significantly higher in patients with prior hemorrhage (p < 0.04). Preexisting neurologic dysfunction improved/dissolved in 50% and remained stable in 45%. CONCLUSIONS: Large AVMs need a long time period to obliterate and show a high bleeding risk. Multimodal treatment strategies are required to reduce treatment volume before radiotherapy. PMID- 16376489 TI - French multicenter phase III randomized study testing concurrent twice-a-day radiotherapy and cisplatin/5-fluorouracil chemotherapy (BiRCF) in unresectable pharyngeal carcinoma: Results at 2 years (FNCLCC-GORTEC). AB - BACKGROUND: Unresectable carcinomas of the oropharynx and hypopharynx still have a poor long-term prognosis. Following a previous phase II study, this phase III multicenter trial was conducted between November 1997 and March 2002. METHODS: Nontreated, strictly unresectable cases were eligible. Twice-daily radiation: two fractions of 1.2 Gy/day, 5 days per week, with no split (D1-->D46). Total tumor doses: 80.4 Gy/46 day (oropharynx), 75.6 Gy/44 day (hypopharynx). Chemotherapy (arm B): Cisplatin 100 mg/m2 (D1, D22, D43); 5FU, continuous infusion (D1-->D5), 750 mg/m2/day cycle 1; 430 mg/m2/day cycles 2 and 3. RESULTS: A total of 163 evaluable patients. Grade 3-4 acute mucositis 82.6% arm B/69.5% arm A (NS); Grade 3-4 neutropenia 33.3% arm B/2.4% arm A (p < 0.05). Enteral nutrition through gastrostomy tube was more frequent in arm B before treatment and at 6 months (p < 0.01). At 24 months, overall survival (OS), disease-free survival (DFS), and specific survival (SS) were significantly better in arm B. OS: 37.8% arm B vs. 20.1% arm A (p = 0.038); DFS: 48.2% vs. 25.2% (p = 0.002); SS: 44.5% vs. 30.2% (p = 0.021). No significant difference between the two arms in the amount of side effects at 1 and 2 years. CONCLUSION: For these unresectable cases, chemoradiation provides better outcome than radiation alone, even with an "aggressive" dose-intensity radiotherapy schedule. PMID- 16376490 TI - Outcome of hyperfractionated radiotherapy in chemotherapy-resistant non-Hodgkin's lymphoma. AB - PURPOSE: Patients with chemotherapy-resistant lymphoma have rapidly progressive disease and a poor prognosis. Local symptoms are treated with radiotherapy (RT) for local control. We have reviewed local control and toxicity in patients treated with hyperfractionated accelerated RT. METHODS AND MATERIALS: A total of 34 patients received hyperfractionated RT between 1997 and 2003. The radiation dose was 39.9-40.5 Gy in 30 fractions. The median treatment time was 22 days with twice-daily involved-field RT. The median follow-up was 4.4 years. Response was assessed <3 months after RT and was classified as a complete response, a complete response-unconfirmed, a partial response, or no response. Local control was defined as maintenance of local complete response, complete response-unconfirmed, or lack of local progression with a partial response. Recurrence or progression outside the RT volume was regarded as distant disease. RESULTS: The median age was 53 years; 20 patients were men and 14 were women. The initial diagnosis was Stage I-II in 56% and Stage III-IV in 44%. The disease bulk was > or =10 cm in 35% (n = 12). The histologic features at diagnosis were follicular in 11 (Grade 1 in 4, Grade 2 in 3, and Grade 3 in 4), diffuse large B-cell in 14, peripheral T cell lymphoma in 2, Burkitt-like in 1, mantle cell in 2, natural killer cell in 2, plasmacytoma/lymphoma in 1, and T-cell lymphoblastic in 1. The initial treatment was chemotherapy in 32 patients (94%); 71% were refractory to initial chemotherapy and 29% developed a relapse after an initial response. The RT response was complete in 24% (n = 8), complete, unconfirmed in 26% (n = 9), partial in 47% (n = 16), and none in 3% (n = 1). The local control rate was 73% at 1, 2, and 3 years. Grade 1 dermatitis was the most common side effect. CONCLUSION: Hyperfractionated RT provided good local control and was well tolerated. This encouraging result requires additional study with comparison to conventional fractionation regimens. PMID- 16376491 TI - Prospective trial of radiotherapy for patients 80 years of age or older with squamous cell carcinoma of the thoracic esophagus. AB - PURPOSE: To assess the safety and efficacy of external beam radiotherapy for elderly patients with esophageal cancer. METHODS AND MATERIALS: A trial testing external beam radiotherapy (66 Gy within 6.5 weeks) as a single-modality treatment was performed for biopsy-proven squamous cell carcinoma of the thoracic esophagus clinically staged as Stage I and IIA (T1-T3N0M0, International Union Against Cancer, 1987) in patients aged > or =80 years. RESULTS: From January 1999 through December 2002, 51 evaluable patients (35 men and 16 women) with a median age of 83 years (range, 80-91 years) were enrolled from 22 institutions. Of the 51 patients, 18 (35%) had Stage T1 and 33 (65%) had Stage T2-T3 disease. Radiotherapy could be completed in 47 patients (92%) within 43-58 days (median, 49). The actuarial incidence of Grade 3 or worse cardiopulmonary complications at 3 years was 26%, with 3 early deaths, and correlated significantly with the size of the anteroposterior radiotherapy portals. The median survival time and overall survival rate at 3 years was 30 months and 39% (95% confidence interval, 25-52%), respectively. CONCLUSION: The results of high-dose radiotherapy in octogenarians are comparable to those in younger patients, but meticulous treatment planning and quality control is required. PMID- 16376492 TI - Conservation treatment of the eye: Conformal proton reirradiation for recurrent uveal melanoma. AB - PURPOSE: To evaluate the outcomes of a second course of proton beam radiation therapy (PBRT) in patients with recurrent uveal melanoma. METHODS AND MATERIALS: Thirty-one patients received a second course of PBRT. The mean interval between the first and the second PBRT course was 50.2 months (range, 8-165 months). Most patients (87%) received 70 cobalt Gray equivalent (CGE) for both courses. Visual acuity was 20/200 or better in 30 patients initially and in 22 patients at the second treatment. The mean follow-up time after the second treatment was 50 months (range, 6-164 months). RESULTS: At the time of the last follow-up, 20 patients were classified as having no evidence of disease, defined as tumor regression or an absence of tumor progression. Nine eyes (29%) were enucleated because of local recurrence (n = 5) or intractable pain (n = 4). The 5-year eye retention rate was 55% (95% confidence interval: 25.2-77.4). Six of the 22 patients who retained the eye (27%) had useful vision (20/200 or better). CONCLUSIONS: A second course of PBRT for recurrent uveal melanoma to total doses between 118 and 140 CGE was associated with a relatively good probability of local control and a low enucleation rate. Although most patients lost vision, the majority were able to retain the reirradiated eye. Further evaluation is needed to assess metastasis-free survival of additional proton irradiation vs. enucleation after local recurrence. PMID- 16376493 TI - Prevention of urinary tract infections in palliative radiation for vertebral metastasis and spinal compression: a pilot study in 71 patients. AB - PURPOSE: To assess the impact of bladder instillations of hyaluronic acid (HA) on the prevalence of urinary tract infection (UTI) in patients receiving emergency radiotherapy for metastatic spinal cord compression. METHODS AND MATERIALS: Patients were recruited consecutively at one center and assigned to usual care (UC) (n = 34, mean age 62.2 years) or UC with once-weekly HA instillation (UC + HA) (Cystistat: 40 mg in 50 mL phosphate-buffered saline) (n = 37; mean age, 63.1 years). All patients had an indwelling catheter and received radiotherapy. UTI status was assessed at baseline and during hospitalization. RESULTS: At baseline, patient groups were comparable, except for the prevalence of UTI at baseline, which was 11.8% and 0% in the UC and UC + HA patients, respectively (p = 0.0477). During hospitalization, 76.5% (vs. 11.8% at baseline, p < 0.0001) of the UC patients had a UTI compared with 13.5% (vs. 0% at baseline, p = 0.0541) of the UC + HA patients (p < 0.0001). Both groups were hospitalized for similar periods (19.8 days [UC] vs. 18.5 days, p = 0.4769) and received equivalent radiotherapy sessions (4.6 [UC] vs. 5.8 sessions, p = 0.2368). CONCLUSIONS: Patients receiving UC + HA had a 5.7-fold decrease in UTI prevalence over the hospitalization period compared to UC patients, suggesting that bladder instillations of HA effectively prevent UTI in patients with indwelling catheters receiving radiotherapy for nerve compression. PMID- 16376494 TI - X-ray volume imaging in bladder radiotherapy verification. AB - PURPOSE: To assess the clinical utility of X-ray volume imaging (XVI) for verification of bladder radiotherapy and to quantify geometric error in bladder radiotherapy delivery. METHODS AND MATERIALS: Twenty subjects undergoing conformal bladder radiotherapy were recruited. X-ray volume images and electronic portal images (EPIs) were acquired for the first 5 fractions and then once weekly. X-ray volume images were co-registered with the planning computed tomography scan and clinical target volume coverage assessed in three dimensions (3D). Interfraction bladder volume change was described by quantifying changes in bladder volume with time. Bony setup errors were compared from both XVI and EPI. RESULTS: The bladder boundary was clearly visible on coronal XVI views in nearly all images, allowing accurate 3D treatment verification. In 93.5% of imaged fractions, the clinical target volume was within the planning target volume. Most subjects displayed consistent bladder volumes, but 25% displayed changes that could be predicted from the first three XVIs. Bony setup errors were similar whether calculated from XVI or EPI. CONCLUSIONS: Coronal XVI can be used to verify 3D bladder radiotherapy delivery. Image-guided interventions to reduce geographic miss and normal tissue toxicity are feasible with this technology. PMID- 16376495 TI - A novel analytical ELISA-based methodology for pharmacologically active saikosaponins. AB - A competitive enzyme-linked immunosorbent assay (ELISA) for saikosaponins was established using monoclonal antibody (MAb) 3G10. Hybridoma 3G10 prepared by fusing splenocytes immunized with saikosaponin a-BSA (SSa-BSA) conjugate and a hypoxanthine-aminopterin-thymidine (HAT)-sensitive mouse myeloma cell line, P3 X63-Ag8-U1, secreted monoclonal antibodies with wide cross-reactivity to saikosaponins including saikosaponin b(2) (SSb(2)), c (SSc) and d (SSd), which are stereo and/or regio isomers of SSa. The method, at an effective measuring range of 0.6 mug /ml to 2.3 mug/ml of SSa, successfully detected total saikosaponins in Bupleuri radix and Kampo medicines prescribed with Bupleuri radix. Good correlation between ELISA and HPLC analyses of total saikosaponin in a crude extract of Bupleuri radix was obtained after hydrolysis of acyl saikosaponins by treatment with a mild alkaline solution. PMID- 16376496 TI - Medicinal potential from in vivo and acclimatized plants of Cleome rosea. AB - Methanolic extracts obtained from different organs of Cleome rosea, collected from its natural habitat and from in vitro-propagated plants, were submitted to in vitro biological assays. Inhibition of nitric oxide (NO) production by J774 macrophages and antioxidant effects by protecting the plasmid DNA from the SnCl(2)-induced damage were evaluated. Extracts from the stem of both origins and leaf of natural plants inhibited NO production. The plasmid DNA strand breaks induced by SnCl(2) were reduced by extracts from either leaf or stem of both sources. On the other hand, root extracts did not show any kind of effects on plasmid DNA, and presented significant toxic effects to J774 cells. The results showed that C. rosea presents medicinal potential and that the acclimatization process reduces the plant toxicity both to plasmid DNA and to J774 cells, suggesting the use of biotechnology tools to obtain elite plants as source of botanical material for pharmacological and phytochemical studies. PMID- 16376497 TI - Transposable elements as a significant source of transcription regulating signals. AB - Transposable elements (TEs) are major components of eukaryotic genomes, contributing about 50% to the size of mammalian genomes. TEs serve as recombination hot spots and may acquire specific cellular functions, such as controlling protein translation and gene transcription. The latter is the subject of the analysis presented. We scanned TE sequences located in promoter regions of all annotated genes in the human genome for their content in potential transcription regulating signals. All investigated signals are likely to be over represented in at least one TE class, which shows that TEs have an important potential to contribute to pre-transcriptional gene regulation, especially by moving transcriptional signals within the genome and thus potentially leading to new gene expression patterns. We also found that some TE classes are more likely than others to carry transcription regulating signals, which can explain why they have different retention rates in regions neighboring genes. PMID- 16376498 TI - Whole genome computational comparative genomics: A fruitful approach for ascertaining Alu insertion polymorphisms. AB - Alu elements are the most active and predominant type of short interspersed elements (SINEs) in the human genome. Recently inserted polymorphic (for presence/absence) Alu elements contribute to genome diversity among different human populations, and they are useful genetic markers for population genetic studies. The objective of this study is to identify polymorphic Alu insertions through an in silico comparative genomics approach and to analyze their distribution pattern throughout the human genome. By computationally comparing the public and Celera sequence assemblies of the human genome, we identified a total of 800 polymorphic Alu elements. We used polymerase chain reaction-based assays to screen a randomly selected set of 16 of these 800 Alu insertion polymorphisms using a human diversity panel to demonstrate the efficiency of our approach. Based on sequence analysis of the 800 Alu polymorphisms, we report three new Alu subfamilies, Ya3, Ya4b, and Yb11, with Yb11 being the smallest known Alu subfamily. Analysis of retrotransposition activity revealed Yb11, Ya8, Ya5, Yb9, and Yb8 as the most active Alu subfamilies and the maintenance of a very low level of retrotransposition activity or recent gene conversion events involving S subfamilies. The 800 polymorphic Alu insertions are characterized by the presence of target site duplications (TSDs) and longer than average polyA tail length. Their pre-integration sites largely follow an extended "NT-AARA" motif. Among chromosomes, the density of Alu insertion polymorphisms is positively correlated with the Alu-site availability and is inversely correlated with the densities of older Alu elements and genes. PMID- 16376499 TI - Characterization of kynurenine aminotransferase III, a novel member of a phylogenetically conserved KAT family. AB - Kynurenine aminotransferase (KAT) is an enzyme responsible for synthesis of kynurenic acid (KYNA), a well established neuroprotective and anticonvulsant agent, involved in synaptic transmission and implicated in the pathophysiology of schizophrenia, Huntington's disease and other neurological disorders. We have shown previously that kat2-/- mice had lower hippocampal KYNA levels and were more hyperactive than wild-type mice. However, these abnormalities occur early and are transitory coinciding with restoration of KYNA levels, suggesting that compensatory changes or ontogenetic expression of another unknown homolog may account for the normalization of KYNA levels in the adult kat2-/- mice brain. Here, we report the isolation of a novel KAT molecule, kat3, from mouse and human brain cDNA libraries. The encoded 454 amino acids of human KAT III share 64.8% similarity to that of KAT I and 30.1% to KAT II. Northern blot analysis demonstrated that kat3 mRNA is widely expressed but with higher expression levels in liver, kidney, heart, and neuroendocrine tissues. RT-PCR and Northern analysis showed that kat3 expression starts as early as postnatal day (PND) 7 and peaks in adult. The mRNA level of kat3 and kat1 when measured together is significantly higher at PND 60 in kat2-/- mice than those of wild-type mice indicating possible co-regulation of expression levels. RNA-interference (RNAi) directed towards transcripts for either R03A10.4 or F28H6.3 in Caenorhabditis elegans which are kat1 and kat3 orthologs, respectively, did not result in any gross abnormalities. Our results show that upregulation of kat3 and kat1 may be responsible for the phenotypic rescue on kat2-/- mice. PMID- 16376500 TI - A keratin 18 transgenic zebrafish Tg(k18(2.9):RFP) treated with inorganic arsenite reveals visible overproliferation of epithelial cells. AB - Inorganic arsenic has strong human carcinogenic potential, but the availability of an animal model to study toxicity is extremely limited. Here, we used the transgenic zebrafish line Tg(k18(2.9):RFP) as an animal model to study arsenite toxicity. This line was chosen because the red fluorescent protein (RFP) is expressed in stratified epithelia (including skin), due to the RFP reporter driven by the promoter of the zebrafish keratin 18 gene. We titrated doses of inorganic arsenite for zebrafish embryos and found that arsenite exposure at 50 microM for 120 h was suitable for mimicking a long-term, chronic effect. When embryos derived from Tg(k18(2.9):RFP) adults were treated with this arsenite dose and time of exposure, abnormal phenotypes were not noticeable under the light microscope. However, arsenic keratosis was visible in the epithelial cells under the fluorescent microscope. Morphological defects became more severe with increased dose and exposure duration, suggesting that the severity of skin lesions was dose- and time-dependent. Histochemical examination of keratosis after 4',6'-diamidino-2-phenylindole hydrochloride (DAPI) staining showed that the epithelial cells overproliferated after treatment with arsenite. Therefore, this Tg(k18(2.9):RFP) zebrafish line is an excellent model for studying toxicity induced by inorganic arsenite and may have potential for studying other environmental pollutants. PMID- 16376501 TI - Solubilization and quantification of lycopene in aqueous media in the form of cyclodextrin binary systems. AB - An optimized kneading method for the preparation of lycopene-cyclodextrin binary systems was developed leading to solubilization of lycopene in water and 5% (w/v) dextrose solution. Lycopene quantification in the prepared binary systems was performed by a developed spectrometric method that followed a successful single step extraction with dichloromethane. Storage stability characteristics of the binary systems were studied at 4 degrees C in solution and at -20 degrees C in the lyophilized products. Lycopene content was monitored at lambda(max)=482 nm, the limit of detection was 0.41 microg/ml and relative standard deviation was less than 3.1%. The results obtained with the spectrometric method were confirmed by a HPLC method. In the presence of cyclodextrins, lycopene concentration in water was 8.0+/-1.0, 27.1+/-3.2 and 16.0+/-2.2 microg/ml for beta-CD, HP-beta-CD and Me-beta-CD, respectively. In 5% (w/v) aqueous dextrose solutions the corresponding values were 16.0+/-1.8, 48.0+/-5.1 and 4.0+/-0.5 microg/ml, respectively. At 4 degrees C, storage stability of lycopene-cyclodextrin binary systems in water or 5% (w/v) aqueous dextrose solutions, was limited (t(1/2)=1-4 days). Addition of the antioxidant sodium metabisulfite increased the stability of lycopene-HP-beta-CD binary system in water. At -20 degrees C, the lyophilized lycopene-cyclodextrin binary systems were stable for at least 2 weeks. PMID- 16376502 TI - Remarks on: "Paternity analysis in special fatherless cases without direct testing of alleged father" [Forensic Science International 146S (2004) S159 S161]. PMID- 16376503 TI - Dermal lesions after post mortem petrol-exposure. AB - The effect of post mortem contact of the skin with petrol was investigated in 18 corpses with exposure times between 10 min and 24 h. The earliest onset of skin changes was observed within 2 h of exposure. They consisted of swelling and wrinkling with detachment of the upper layers of the skin (positive Nikolski's sign). Histologically the lesions appeared as non-vital acantholyses located in the prickle-cell layer with formation of intra-epidermal bullae. An influence of age and sex could be excluded, the earlier onset of lesions at 22 degrees C than at 4 degrees C was not statistically significant. These findings prove that post mortem petrol exposure may lead to dermal lesions. PMID- 16376504 TI - Small cell carcinoma of the larynx treated with irinotecan and cisplatin. AB - We report a case of advanced small cell carcinoma in the larynx, which was treated with Irinotecan hydrochloride (CPT-11) chemotherapy. The patient was free of disease for 4 years after treatment. Several chemotherapeutic agents for small cell carcinoma have been proposed; however, median survival time has been miserable, especially in advanced cases. For the cure of the aggressive lethal behavior of this disease, chemotherapy with CPT-11 might be effective to improve median survival of patients with small cell carcinomas of the larynx. PMID- 16376505 TI - Early-onset absence epilepsy: clinical and electroencephalographic features in three children. AB - To describe the clinical and electroencephalographic features of three infants diagnosed as having early-onset absence seizures. Two males and one female, aged 21-29 months were seen in our neuropaediatric outpatient clinic because of daily episodes of motor arrest and loss of contact. Neurological examination and mental development was considered normal in all of them. Two out three had first-degree relatives with seizures with onset in the childhood and favourable evolution in the adulthood. A video-electroencephalogram was requested. Ictal EEG revealed a normal background and generalised spike-and-wave complexes at 3-3.5 Hz accompanied by disruption of ongoing activity in keeping with absence seizures. The duration of seizures ranged from 2 to 10s. One child (patient 2) experienced rhythmic myoclonic jerks in upper limbs and head as those described in myoclonic absences. Clinical and electroencephalographic follow-up ranged from 8 months to 4 years. Two children were on treatment with valproate and in the case of the patient 3, the combination of valproate and ethosuximide was necessary. Control of absence seizures was achieved in all our cases. Absence seizures should be considered as a possible cause of transient impairment of consciousness even among infants minor than 3 years of age. A video-electroencephalogram is the method of choice in the diagnostic evaluation and syndromic classification of these paroxysmal events. PMID- 16376506 TI - Aberrant peroxisome morphology in peroxisomal beta-oxidation enzyme deficiencies. AB - Peroxisomes are ubiquitous organelles in eukaryotic cells and surrounded by a single membrane, and undergo considerable changes in size, shape and number. Peroxisomal disorders are classified into two categories: peroxisome biogenesis disorders (PBDs) and single-enzyme deficiencies (SEDs). Morphologically aberrant peroxisomes called 'peroxisomal ghosts' in PBDs are well known, however, a morphological approach to the study of peroxisomes in SEDs has been rarely reported. Here, we investigated the morphology of peroxisomes in cultured fibroblasts from patients lacking peroxisomal beta-oxidation enzymes, including acyl-CoA oxidase (AOX) or D-3-hydroxyacyl-CoA dehydratase/D-3-hydroxyacyl-CoA dehydrogenase bifunctional protein (D-BP). Morphological analysis by immunofluorescence examination using an antibody against catalase revealed a smaller number of large peroxisomes in fibroblasts from these patients. Moreover, immunoelectron microscopy using an antibody against the 70-kDa peroxisomal membrane protein (PMP70) showed large peroxisomes with various horseshoe-shaped membrane structures. These results give an important clue to elucidating the division of peroxisomes and how peroxisomes change in size, shape, number and position within cells, which are subjects for future study. PMID- 16376507 TI - A distinct autosomal recessive ataxia maps to chromosome 12 in an inbred family from Jordan. AB - Autosomal recessive ataxias are a heterogeneous group of rare disorders characterized by early onset ataxia associated with neurologic, ophthalmologic or systemic signs. The ataxias associated with myoclonus, epilepsy and progressive neurological degeneration are usually included with the progressive myoclonus epilepsies, one of which is Unverricht-Lundborg disease. We identified four siblings with ataxia, juvenile onset progressive action tremor and atonic seizures from a Jordanian family. The mode of inheritance of this syndrome is autosomal recessive. We performed a genome-wide screen for linkage and fine mapped the region that contains the disease locus. The four affected siblings have ataxia noted at the onset of walking with dysarthria and bulbar features, but no cerebellar hypoplasia on MRI. They all developed a fine tremor that progressed to a coarse action tremor, as well as atonic seizures. Treatment with valproate fully controlled the seizures and improved the tremor, but did not change the course of the ataxia. We mapped the gene responsible for this disorder to the pericentromeric region of chromosome 12. A recently described autosomal recessive variant of Unverricht-Lundborg disease also maps to the same region. We discuss the similarities and differences between our family and the family with the Unverricht-Lundborg disease variant. PMID- 16376508 TI - Treatment of epilepsy with electrical status epilepticus during slow sleep and its related disorders. AB - To elucidate an effective therapeutic strategy for 'ESES syndrome', epilepsy with electrical status epilepticus during slow sleep (ESES) and its related epileptic disorders, we studied the effect of treatment on the EEG pattern of continuous spike-waves during slow wave sleep (CSWS) in 15 afflicted patients. Basically performed in the following order, the employed therapies included (1) high-dose valproate (VPA) therapy (serum level >100 microg/ml); (2) a combination therapy of VPA and ethosuximide (ESM); (3) short cycles of high-dose diazepam (oral or intrarectal DZP, 0.5-1 mg/kg per day for 6-7 days); and (4) intramuscular synthetic ACTH-Z therapy (0.01-0.04 mg/kg per day for 11-43 days). Regarding the initial EEG effect, a remission of CSWS was achieved by high-dose VPA therapy in 7 of 15 trials (47%), by the combination therapy of VPA and ESM in 3/7 trials (43%), by short cycles of high-dose DZP in 2/4 trials (50%), and by ACTH-Z therapy in 2/5 trials (40%). A permanent remission of ESES syndrome was achieved by high-dose VPA therapy and/or combination therapy of VPA and ESM in 10 patients (67%). The effects of short cycles of high-dose DZP and ACTH-Z therapy were at best temporary. Our strategy for the treatment of ESES syndrome is therefore considered valid. PMID- 16376509 TI - Cerebrovascular disease and varicella in children. AB - Varicella-associated stroke has been reported with increasing frequency in recent years. In many cases, diagnosis is difficult because of the late onset of manifestations after the acute infectious episode. Four cases of cerebrovascular disease after varicella infection were observed. Three children presented hemiparesis and one facial paresis. The neuroradiological findings comprised stenosis/occlusion of middle cerebral artery or nucleo capsular signal alteration. Because, several pathogenetic mechanisms have been proposed as the cause of stroke, the relationship between prothrombotic conditions, antipospholipid antibodies and stroke in these patients is discussed. The difficulty in defining the pathogenesis of the ischemic episode is related to problems in the choice of antithrombotic treatment, which is still not standardized and must be decided on individual basis. In the event of rapid onset of stroke after exanthem high dose antiviral therapy seems to be justified. On the basis of our experience and of literature data on varicella-associated stroke, we recommend that VZV infection be taken into account in every episode of stroke in children. PMID- 16376510 TI - MECP2 mutations are an infrequent cause of mental retardation associated with neurological problems in male patients. AB - Mutations in the methyl-CpG-binding protein 2 (MECP2) gene located on Xq28, cause Rett syndrome (RTT) in female patients. Meanwhile, nonmosaic MECP2 mutations unknown in girls have been found in an increasing number of male patients with a normal 46, XY karyotype. They can cause a broad spectrum of neurodevelopmental disorders which often show a combination of mental retardation (MR) with neurological symptoms. We present the results of MECP2 analysis in a group of 72 male patients with an unexplained combination of MR and neurological features, and review the mutational reports published on male patients since the discovery of the MECP2 gene. Analysis included sequencing of exon 1 which thus far was mostly omitted from DNA screening. One pathogenic mutation has been found in a patient with Rett variant, in addition to an unclassified variant and a series of nonpathogenic changes. No changes have been found in exon 1. Criteria for testing of male patients are classic RTT, severe neonatal encephalopathy, and RTT variant which may be clinically underrecognized. Testing can also be considered in males with a combination of unexplained MR and (progressive) neurological manifestations although the yield of MECP2 analysis is probably low in this situation. Based on the literature, MECP2 testing in males with MR only is debatable. PMID- 16376511 TI - A novel R275X mutation of the SLC25A15 gene in a Japanese patient with the HHH syndrome. AB - The hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome (MIM 238970) is an autosomal recessive metabolic disorder caused by a deficiency of the mitochondrial ornithine transporter, one of the urea cycle components. Mutations in the SLC25A15 gene have been coupled to the HHH syndrome. We describe a Japanese female patient with the HHH syndrome due to a novel homozygous R275X SLC25A15 mutation and male sibling who presumably carried the same mutation. He exhibited slowly progressive deterioration with seizures, a gait disturbance due to polyneuropathy, episodic confusion, and died of acute encephalopathy at 34 years of age while the proband exhibited moderate mental retardation, seizures, mild spastic paraplegia, and deafness without neurological deterioration for more than 20 years. The clinical features of previously documented patients with the homozygous SLC25A15 mutation demonstrated that genotype did not simply correlate with clinical severity. The phenotypic variability might depend on other factors, such as dietary and other genetic ones. PMID- 16376512 TI - A novel splicing mutation of the ATRX gene in ATR-X syndrome. AB - X-linked alpha-thalassemia/mental retardation syndrome (ATR-X, MIM#301040) is an X-linked recessive condition affecting males. ATR-X is characterized by severe mental retardation, mild HbH disease, dysmorphic facies, and genital and skeletal abnormalities. ATR-X is caused by mutations in the ATRX gene. Most mutations affect two functionally important domains, the ADD domain and the helicase domain. Here, we report on two brothers with the ATR-X phenotype without HbH disease; both had a mutation in the 5' upstream region of the ADD domain of the ATRX gene. This mutation was a G to T nucleotide substitution at the 3' end of exon 5 and resulted in splicing out of exons 5 and 6. Analysis of cDNA structure may clarify genotype-phenotype correlations in ATR-X because splicing mutation could be detectable only by cDNA analysis. PMID- 16376513 TI - Subacute sclerosing panencephalitis presenting with Balint's syndrome. AB - Visual-spatial agnosis, praxis deficits and hallucinations can be the features of subacute sclerosing panencephalitis (SSPE) in the early period. This study describes a 15-year-old boy with SSPE presenting with visual agnosia, prosopagnosia, simultanagnosia, optic ataxia, and oculomotor apraxia which are compatible with Balint syndrome. MRI revealed heterogenous and abnormal signal changes in the bilateral parieto-occipital areas. The signals were more prominent on the left. The case discussed herein is important because he was diagnosed as Balint's syndrome by means of clinical and neuroradiological findings before the onset of known symptomatology as dementia/myoclonus of SSPE. The case is also illustrative of the need to emphasize the prompt evaluation of a patient's cortical features before clinical progression becomes apparent. This evaluation should be performed in the early period if correct diagnosis is to be reached. PMID- 16376514 TI - Ethylmalonic encephalopathy-report of two cases. AB - Ethylmalonic encephalopathy is a rare metabolic disease presenting in infancy with developmental delay, acrocyanosis, petechiae, chronic diarrhea and early death. The biochemical characteristics of this autosomal recessive disease are urinary organic acid abnormalities. Recently it has been found to be caused by mutations in the ETHE1 gene, located on Ch19q13. Only about 30 patients have been reported, and we describe two additional cases. The first patient showed a typical clinical picture and biochemical abnormalities, with additional atypical clinical features. Neuroimaging studies showed extensive changes. A new homozygous mutation in exon 3 of the ETHE1 gene was found. The second patient was not investigated genetically; however besides the typical clinical picture and biochemical profile he was found to have cytochrome C oxidase deficiency. PMID- 16376515 TI - Sentinel node mapping in colon carcinoma: in-vivo versus ex-vivo approach. AB - AIM: The aim of this study was to determine if ex-vivo and in-vivo technique of lymphatic mapping for colorectal cancer (CCR) result in similar sentinel lymph node (SLN) identification and accuracy rates. METHODS: Thirty consecutive patients with 32 CCR underwent in vivo SLN mapping. After completion of the colectomy, we remapped the SLN in the operative specimens from patients who had undergone successful in vivo lymphatic mapping. RESULTS: At least one SLN was identified by in vivo approach in 32 tumours. 1.5 SLNs (1-3) and 1.8 SLNs (1-4) (p=0.24) were identified by the in vivo and the ex vivo technique, respectively. All SLNs identified by the in vivo technique were also identified by the ex vivo technique. In six cases one and in two cases two additional SLNs were identified with the ex vivo technique. Twelve percent of tumours were upstaged. CONCLUSION: Ex vivo SLN mapping is as accurate as the in vivo technique in defining SLN and does have the ability to upstage some patients with CCR. The ex vivo technique could be used either as a primary lymphatic mapping procedure or secondarily for failed in vivo attempts at lymphatic mapping. PMID- 16376516 TI - Genomic organization of the rat Clock gene and sequence analysis in inbred rat strains. AB - While mutations in genes that function in the core molecular clock may disrupt circadian periodicity, their relevance to diurnal variation in metabolic, cardiovascular, and respiratory function is unknown. The circadian Clock gene product is an essential regulator of central and peripheral circadian rhythms in mammals. We have elucidated the complete exon-intron organization of the Clock gene in rat and have carried out an extensive search for single nucleotide polymorphisms (SNPs) in a panel of 12 inbred rat strains that exhibit diversity in studies of central and peripheral organ function and disease. The rat Clock gene consists of 23 exons spanning approximately 75 kb. Comparative sequence analysis identified 33 novel SNPs, including 32 that distinguish the Brown Norway (BN) rat from the other strains studied. Most notable were two novel mutations in the BN sequence at exon 8, Ile131Val and Ile132Val, occurring in a segment of the highly conserved PAS-A domain of the protein. These results afford the opportunity to assess the impact of genetic variation in Clock on central and peripheral functions subject to the core molecular clock and to test the importance of Clock variants in explaining diversity among rat strains in the expression of phenotypes, such as blood pressure, subject to circadian oscillation. PMID- 16376517 TI - Comparative study of the linkage disequilibrium of an ENCODE region, chromosome 7p15, in Korean, Japanese, and Han Chinese samples. AB - The extent and pattern of linkage disequilibrium (LD) in the human genome provide important information for disease gene mapping. Previous studies have shown that LDs vary depending on chromosomal regions and populations. As the Asian samples of the International HapMap Project consisted of Japanese and Chinese populations, it was of interest whether we could use the HapMap data as a reference to carry out association studies of common complex diseases in a closely related population, such as Koreans. We have compared the LD and recombination patterns defined by single-nucleotide polymorphisms (SNPs) in ENCODE region ENm010, chromosome 7p15.2, in Korean, Japanese, and Chinese samples and further tested the robustness of tagSNPs among the Asian samples. We genotyped 792 SNPs in 500 kb (chromosome 7: 26699793-27199792, NCBI build 34) from 90 unrelated Koreans by fluorescence polarization detection and compared the data with Asian data from the HapMap project. Despite some differences in the position of high LD region boundaries, the overall patterns of LD were remarkably similar across the three samples, reflecting strong genetic affinities among them. Furthermore, the haplotype tag SNP transferability across the three samples was greater than 90%. Our results support the initial suggestion that the populations genotyped in the HapMap project might serve as reference populations for the selection of tagSNPs in association studies. PMID- 16376518 TI - Hostility and pain are related to inflammation in older adults. AB - Chronically elevated systemic inflammation has a dramatic impact on health for older individuals. As stress-related responses, both hostility and pain perception may contribute to inflammation which in turn may maintain negative emotion and pain over time. We used structural equation modeling to examine the degree to which trait hostility and pain were uniquely associated with C-reactive protein (CRP) and serum IL-6 levels over a 6-year span in a sample of older adults. The sample included 113 present or former caregivers of a spouse with dementia and 101 non-caregivers. After accounting for depression, health behaviours, and other risk factors, which were also assessed longitudinally, pain and, to a lesser extent, hostility were uniquely associated with plasma levels of CRP but not IL-6. When examined separately, the association between pain and CRP was significant only for caregivers, while the association between hostility and CRP was comparable for the two groups. These findings suggest that hostility may play a role in a cycle of inflammation among older adults, and that pain may be particularly problematic for those under chronic stress. Our results also shed light on inflammation as a mechanism underlying the effects of hostility on cardiovascular disease morbidity and mortality. PMID- 16376519 TI - Gender difference in basal and stress levels of peripheral blood leukocytes in laboratory rats. AB - The aim of the present study was to investigate gender differences in numbers and function of blood immune cells in stressed and non-stressed laboratory rats. Psychosocial stress in adult male or female rats was induced by social confrontation of an intruder rat with a resident opponent for 2 h. Behavioral analysis indicated that intruders of both sexes were clearly defeated and had markedly higher plasma corticosterone concentrations than unstressed home cage controls at the end of the confrontation. Lower numbers of CD4, CD8, and B cells as well as a reduced proliferative response of lymphocytes to ConA were observed in stressed groups of either sex. However, some important gender differences were also observed. Stressed males had higher granulocyte numbers than controls, while granulocyte numbers remained unchanged in stressed females. Similarly, stressed males had higher phagocytic activity than stressed females. Second, there was a gender difference in some basal values. Female controls had lower NK cell numbers than control males. Interestingly, NK numbers in stressed males decreased considerably, reaching the same low levels as in (stressed and control) females. In addition, females exhibited higher basal corticosterone concentrations than males. To summarise, these data do not indicate a superior blood cellular immune function in female rats, neither for the control nor the stress condition. However, the data clearly suggest that male and female rats should not be considered as a uniform group with respect to their immunological response to stress. PMID- 16376520 TI - Regulation of cellular functions by the ERK5 signalling pathway. AB - Extracellular-regulated protein kinase 5 (ERK5) is a mitogen-activated protein kinase (MAPK) regulated by a wide range of mitogens and cellular stresses. Since its cloning in 1995, the lack of biological tools, including antibodies and specific inhibitors, have made it one of the least studied MAPK subfamilies. The discovery that ERK5 was an important contributor to cell survival mechanisms has increased interest in this signalling pathway. The ability of inhibitors of the classical MAPK (ERK1/2) cascade to block ERK5 activation suggested that ERK5 might regulate some cellular functions originally attributed to ERK1/2. For example, ERK5 is suspected to mediate the effects of numerous oncogenes. A link between abnormal levels of ERK5 expression and cancers was established by the analysis of human tumours. Recently, the targeted deletions of the erk5 and the mek5 genes in mice have provided genetic evidence that the ERK5 cascade is a non redundant signalling pathway essential for normal cardiovascular development. The analysis of genetically modified mice in which the erk5 gene can be specifically deleted in certain tissues is shedding light into the physiological function of the ERK5 pathway during development and pathogenesis. PMID- 16376522 TI - Electronically monitored adherence in outpatients with schizophrenia or schizoaffective disorder: a comparison of first- vs. second-generation antipsychotics. AB - OBJECTIVE: To examine the effect of first- vs. second-generation antipsychotics on electronically monitored adherence in outpatients with schizophrenia or schizoaffective disorder. METHOD: The sample consisted of 61 outpatients with a DSM-IV diagnosis of schizophrenia or schizoaffective disorder who took either a first-generation (N=25) or second-generation (N=36) oral antipsychotic during study participation. Treatment group designation (first-vs. second-generation antipsychotic) was based on the particular antipsychotic medication the patient was receiving as part of routine care at study entry (i.e., non-random assignment). Adherence to prescribed antipsychotic medication was assessed monthly over a 6-month study period using electronic monitoring (EM) of medication bottle opening. Various participant characteristics were collected at baseline to test for group differences and for potential associations with prospectively measured adherence. The primary data analysis was a mixed-model analysis of repeated measures. RESULTS: The analysis of EM adherence revealed no significant difference between those taking first-generation (6-month adjusted mean adherence=64.35%) and second-generation antipsychotics (6-month adjusted mean adherence=69.17%; Group effect, p=.29) and no significant Group X Period interaction (p=.13). CONCLUSION: There was no statistical difference in EM adherence, over a 6-month period, between patients taking first- and second generation antipsychotics. However, since the patients were not randomized, conclusions must be interpreted within the context of the quasi-experimental design used in the current study. PMID- 16376523 TI - Extreme life on Earth--past, present and possibly beyond. AB - Life may have been present on Earth since about 3.8 billion years ago or earlier. Multidisciplinary research, especially on the paleobiology and evolution of early microorganisms on Earth and the microbiology of extremophiles in the Earth's environments and under space conditions, enables the defining of strategies for the detection of potential extraterrestrial life by determining biosignatures and the environmental envelope of life. PMID- 16376521 TI - Vitamin C-induced activation of phospholipase D in lung microvascular endothelial cells: regulation by MAP kinases. AB - Our earlier studies have shown that vitamin C at pharmacological doses (mM) induces loss of redox-dependent viability in bovine lung microvascular endothelial cells (BLMVECs) that is mediated by oxidative stress. Therefore, here, we investigated the vitamin C-induced activation of the lipid signaling enzyme, phospholipase D (PLD) in BLMVECs. Monolayer cultures of BLMVECs were treated with vitamin C (0-10 mM) for different time periods (0-2 h) and the activity of PLD was determined. Vitamin C induced activation of PLD in BLMVECs in a time- and dose-dependent fashion that was significantly attenuated by antioxidants, p38 mitogen-activated protein kinase (p38 MAPK)-specific inhibitor (SB203580), extracellular signal-regulated protein kinase (ERK)-specific inhibitor (PD98059), and transient transfection of cells with dominant-negative (DN)-p38 MAPK and DN-ERK1/ERK2. Vitamin C also induced phosphorylation and enhanced the activities of p38 MAPK and ERK in BLMVECs in a time-dependent fashion. It was also evident that vitamin C induced translocation of PLD(1) and PLD(2), association of p38 MAPK and ERK with PLD(1) and PLD(2), threonine phosphorylation of PLD(1) and PLD(2) and SB203580- and PD98059-inhibitable threonine phosphorylation of PLD(1) in BLMVECs. Transient transfection of BLMVECs with DN-p38 MAPK and DN-ERK1/ERK2 resulted in marked attenuation of vitamin C induced phosphorylation of threonine in PLD(1) and PLD(2). We, for the first time, showed that vitamin C at pharmacological doses, activated PLD in the lung microvascular ECs through oxidative stress and MAPK activation. PMID- 16376524 TI - The C-terminus of MinE from Neisseria gonorrhoeae acts as a topological specificity factor by modulating MinD activity in bacterial cell division. AB - MinE regulates the proper placement of the cytokinetic FtsZ ring at midcell by inducing the pole-to-pole movement of MinCD complexes. While the N-terminus of MinE has been implicated in MinD binding, a clear functional role of the C terminus has not been elucidated. We previously determined that MinE from Neisseria gonorrhoeae (Ng) was functional in Escherichia coli (Ec). Thus, using E. coli as a model organism, gonococcal MinE (MinE(Ng)) function was examined by generating amino acid substitutions of highly conserved MinE(Ng) residues and by testing the ability of the mutant proteins to interact with gonococcal MinD (MinD(Ng)), to induce a minicell phenotype upon overexpression, to initiate MinD(Ng) oscillation, and to stimulate MinD(Ng) ATPase activity. N-terminal MinE(Ng) mutants were unable to bind to MinD(Ng); thus, they did not induce a minicell phenotype, promote MinD(Ng) oscillation or stimulate MinD(Ng) ATPase activity. While C-terminal MinE(Ng) mutants exhibited reduced abilities to bind to MinD(Ng), we show that differences in MinD(Ng) binding to the C-terminus of MinE(Ng) alter the ability of MinE(Ng) to properly stimulate MinD(Ng) activity. We present four major findings from our studies of MinE(Ng): both the N- and C termini of MinE(Ng) interact with MinD(Ng); interaction between MinD(Ng) and MinE(Ng) is required for the recruitment of MinD(Ng) to the coiled array; oscillation of MinD(Ng) does not require ATPase stimulation; and, the extent of MinD(Ng) ATPase stimulation depends on the binding strength between MinD(Ng) and the C-terminus of MinE(Ng.). PMID- 16376525 TI - Effects of melatonin and agomelatine in anxiety-related procedures in rats: interaction with diazepam. AB - The anxiolytic potential of melatonin and agomelatine, a potent MT(1/2) receptor agonist, and their combined effects with diazepam, were investigated in rats using the punished drinking test, the safety signal withdrawal operant paradigm, the elevated-plus-maze and hypophagia-induced novelty. In the punished drinking test, evening injections of melatonin (80 mg/kg, IP, but not 20 and 40 mg/kg) and agomelatine (40 mg/kg, IP) increased the number of foot shocks received. However, neither melatonin (40-80 mg/kg) nor agomelatine (20-40 mg/kg) released response suppression during the period associated with the safety signal withdrawal and affected rats' behaviour in the elevated-plus-maze. Furthermore, agomelatine (40 mg/kg) did not enhance food consumption in unfamiliar environment. However, the co-administration of melatonin (80 mg/kg) or agomelatine (20-40 mg/kg) with diazepam, at a dose (0.25 mg/kg) inactive on its own, induced an anxiolytic-like effect in the punished drinking test and the elevated plus-maze. These results indicate that, although mostly devoid of anxiolytic-like action per se, melatonin and agomelatine can potentiate the anxiolytic effects of diazepam. PMID- 16376527 TI - Regulation of drug transporters by PDZ adaptor proteins and nuclear receptors. AB - Drug transporters have been suggested to be involved in various aspects of pharmacokinetics. Identification and characterization of drug transporters have given us a scientific basis for understanding drug disposition, as well as the molecular mechanisms of drug interaction and inter-individual/inter-species differences. On the other hand, regulatory mechanisms of drug transporters are still poorly understood, and information is limited to induction and down regulation of drug transporters by various microsomal enzyme inducers. Little is known about the molecular machinery that directly interacts with the drug transporters. As a first step to clarify such molecular mechanisms, recent studies have identified PDZ (PSD-95/Discs-large/ZO-1) domain-containing proteins that directly interact with the so-called PDZ binding motif located at the C terminus of drug transporters. Some of the PDZ proteins have been suggested to regulate transporters via at least two pathways, i.e. stabilization at the cell surface and direct modulation of transporter function. Therefore, it is possible that membrane transport of therapeutic agents is not only governed by the drug transporters themselves, but also indirectly by PDZ proteins. The PDZ proteins are classified as a family, the members of which are thought to have distinct, but also redundant physiological roles. The purpose of this review article is to summarize the available knowledge on protein interactions and functional modulation of drug transporters. PMID- 16376528 TI - The kinetics and tissue distribution of protein transduction in mice. AB - Protein transduction domains (PTDs) offer an exciting therapeutic opportunity for the treatment of many diseases. An 11-amino acid fragment of human immunodeficiency type 1 (HIV-1) TAT-protein can transduce large, biologically active proteins into mammalian cells; recent evidence has shown an in vivo PTD for the 116 kDa beta-galactosidase protein. However, there is little information on the in vivo distribution of the TAT fusion protein to define the viability of PTDs for human studies. In this study we examined the tissue kinetics and tissue distribution of the PTD-transduced TAT fusion protein in mice. Low (100 microg) or high (500 microg) doses of TAT-beta-galactosidase fusion protein were administrated to mice through four routes (portal vein, i.v., i.p., and oral). Tissues were harvested 15 min, 1h, 6h, 10h, and 24h after treatment. Distribution of beta-galactosidase in various tissues was analysed by in situ staining, enzymatic activity assay, and Western blot analysis. Beta-galactosidase enzyme activity was observed in all tissues (liver, kidney, spleen, lung, bowel, and brain). Beta-galactosidase activity peaked at 15 min in most tissues after portal vein, i.v., and i.p. administration and at 1h after oral dosing in all tissues. Beta-galactosidase activity in the liver at 15 min after portal vein injection (67 milliunits [mU]/mg) was higher than after i.v. (9.8 mU/mg), i.p. (4.4 mU/mg), and oral (0.3 mU/mg) dosing. In situ staining and Western blot results correlated closely with beta-galactosidase enzyme activity assay. The median initial half life for activity was 2.2h, ranging from 1.2h to 3.4h (coefficient of variation=28.9%). The bioavailability of beta-galactosidase activity after an orally administered PTD was 24%. This study details the kinetics and tissue distribution of delivering of a model TAT fusion protein into the mouse via PTD. These data allow rational selection of delivery route and schedules for therapeutic PTD and will aid the use of TAT fusion protein transduction in the development of protein therapies. PMID- 16376529 TI - [Botulism in a neonate]. AB - Botulism was suspected in a 17-day-old breastfed infant who developed over 2 days progressive muscular weakness and hypoventilation. The patient also presented with pupil dilation and light unresponsiveness. The electroencephalogram was normal. Full recovery was obtained after 85 days of artificial ventilation. Diagnosis was confirmed by the presence of the botulin toxin B in the patient serum. The source of the infection was not identified. PMID- 16376530 TI - [Childhood cancer incidence and survival rates in the Rhone-Alpes regional paediatric registry 1987-1999]. AB - Cancer is rare in children, and pediatric malignancies represent only 1% of all cancers. OBJECTIVES: The cure rate is high and increasing, and ongoing data collection is therefore warranted. MATERIALS AND METHODS: Here we report the incidence and survival rates of childhood cancers between 1987 and 1999 in the Rhone-Alpes region of France. RESULTS: A total of 1945 cases were recorded during the study period, with an average of 149.6 new cases per year. The approximate incidence rate was 134.1/10(6) per year and the age-standardized incidence rate was 139.2/10(6) per year. The histological distribution and 5-year survival rates were respectively 30.2 and 73% for leukemia, 12.3 and 91.6% for lymphoma, 24.7 and 60.1% for CNS tumors, 9.1 and 71.1% for neuroblastoma, 2.5 and 94.1% for retinoblastoma, 5.8% and 89.9% for renal tumors, 1 and 75% for liver tumors, 6.1 and 60.9% for bone tumors, 4.1 and 58.6% for soft-tissue tumors, 1.1 and 71% for germ cell tumors, and 2.4 and 85.1% for carcinomas. CONCLUSION: The overall survival rate was 75%. Long-term treatment complications warrant further studies of children who survive into adulthood. PMID- 16376531 TI - Armillaria luteobubalina mycelium develops air pores that conduct oxygen to rhizomorph clusters. AB - Armillaria luteobubalina produces air pores in culture. They consist of two parts: a basal region of tissue elevated to form a mound covered with a rind continuous with that of the colony, but perforated; and an apical region of long parallel hyphae, cemented together by scattered patches of extracellular material. This forms a hydrophobic structure that is elevated above the general level of the mycelial crust and does not easily become waterlogged. Air pores develop near the inoculum plug shortly after inoculation, arising directly from the mycelium, and rhizomorphs are initiated from them. The air pore contains a complex system of gas space connecting the atmosphere with the central canal of each rhizomorph. The tissue beneath the melanised colony crust also contains gas space, especially near air pores. This is also connected with the gas space of each rhizomorph and of each air pore. Measurements with oxygen electrodes show that air pores and their associated rhizomorphs conduct oxygen. The average oxygen conductance of a group of air pores with associated rhizomorphs, within agar blocks, but with rhizomorph apices cut off, was about 700 x 10(-12) m3s(-1), equivalent to about 200 x 10(-12) m3s(-1) for each air-pore. We conclude that the air pores conduct oxygen into the gas space below the pigmented mycelium of the colony, where the rhizomorphs - which also conduct oxygen - originate. A. luteobubalina thus has a complex aerating system which allows efficient diffusion of oxygen into rhizomorphs, and this is likely to facilitate extension of inoculum into low-oxygen environments. PMID- 16376532 TI - Evolution and phylogenetic relationships within Porinaceae (Ostropomycetidae), focusing on foliicolous species. AB - A phylogeny of the lichen family Porinaceae using mitochondrial SSU rDNA sequences is presented, with special focus on foliicolous taxa. Fifty specimens of 28 mostly tropical species, representing eight species groups of Porina as well as the genus Trichothelium, were analysed together with species of other members of Ostropomycetidae, and using Agyriaceae as outgroup. We performed the phylogenetic analyses with a Bayesian approach and under the criterion of maximum parsimony. Four main clades can be distinguished: the P. nitidula-group s. lat. (including Trichothelium, P. papillifera and P. rubescens), the Porina epiphylla group s. lat. (including the P. radiata-, the P. nucula-, the P. imitatrix- and the P. epiphylla-group s. str.) and two clades of the P. rufula-group. The genus Porina as understood by all recent concepts is paraphyletic, and Trichothelium is nested within the Porina nitidula-group. The non-setose P. repanda forms a monophyletic clade with Trichothelium. The tree does not support a monophyletic origin of substrate preferences or photobiont selection. Species-specific associations with morphologically different trentepohlioid photobionts mapped on the tree suggest that closely related mycobiont species switch between different types of algae. PMID- 16376533 TI - Chaetosphaeria tortuosa, the newly discovered teleomorph of Menispora tortuosa, with a key to known Menispora species. AB - Chaetosphaeria tortuosa is described as the newly discovered teleomorph of Menispora tortuosa, based on specimens from Canada and the Czech Republic, and single spore isolations from both morphs. The fungus produces superficial, more or less globose, papillate, dark brown to black smooth perithecia (200-)220-250 x (220-)230-260 microm. The asci are unitunicate, 8-spored, cylindrical-fusiform, (110-)120-133(-145) x 12-14 with a distinct apical, nonamyloid annulus 1-1.5 microm high, 3.5-4 microm wide. The ascospores are fusiform, 19-24 x 5-6 microm, hyaline, 3-septate, smooth, and 2-seriate in the ascus. The morphology of the teleomorph and anamorph are similar to that of C. ovoidea (anamorph: M. glauca), differing in dimensions of asci and ascospores, and in the disposition and morphology of the phialides of the anamorphs. The generic concept and phylogeny of Menispora is briefly discussed, and a key to the 11 species currently accepted in the genus is provided. PMID- 16376534 TI - Septic pulmonary emboli due to periodontal disease. AB - Septic pulmonary embolization (SPE) is a rare but serious disorder. It is a well recognized potential problem in the settings of tricuspid valve endocarditis, septic thrombophlebitis, infected central venous catheters, and postanginal septicemia. Less well documented is the occurrence of SPE in patients with periodontal disease without suppurative thrombophlebitis of the great vessels of the neck. We report a patient with SPE in whom periodontal disease was the only identifiable nidus of infection and review the literature regarding the four other patients reported to have suffered this complication. PMID- 16376535 TI - The use of endobronchial valve device to eliminate air leak. AB - We evaluated an endobronchial valve device in the treatment of surgically created air leak or pneumothorax by eliminating antegrade flow. METHODS: Six sheep underwent general anesthesia with positive pressure ventilation and left thoracotomy. After division of the mediastinal pleura, the contralateral cranial lobe was identified and a 2.5 cmx1.5 cm laceration created with resultant air leak. Using bronchoscopy, we deployed a valve device in the bronchus of the injured segment. Chest drainage tube was placed and the thoracotomy closed. At 1 week (n=3) and 4 weeks (n=3), the animals underwent general anesthesia, bronchoscopy and right thoracotomy. RESULTS: All animals survived the procedure. Bronchoscopic valve device placement in the segmental bronchus resolved the air leak immediately. After closure of thoracotomy, the chest tube demonstrated minimal drainage with no air leak. At 1 and 4 weeks, bronchoscopy showed no change in device location, and the treated segments were atelectatic with fibrous scar at the injured site. CONCLUSIONS: Collapse of a selected lung segment with resolution of air leak can be achieved using bronchoscopically implanted valve device. The valve device may facilitate treatment of patients with post-surgical or post-traumatic persistent air leak. PMID- 16376536 TI - Membrane diffusion in diseases of the pulmonary vasculature. AB - INTRODUCTION: We examined pulmonary diffusing capacity (D(LCO)) and its partition in pulmonary vascular diseases without evident parenchymal disease to assess the pattern and proportionality of change in membrane diffusion (D(m)) and capillary blood volume (V(c)). Disproportionate reduction in D(m) relative to V(c) (low D(m)/V(c)) in these diseases has been attributed to associated alveolar membrane/parenchymal disease, thus providing a potentially important diagnostic tool. METHODS: Diseases included: idiopathic pulmonary arterial hypertension (n=6), chronic thromboembolic disease (n=5), and intravenous drug use (n=14), providing a spectrum of pulmonary vascular diseases. V(c) and D(m) were determined as described by Roughton and Forster. RESULTS: All diseases showed a reduced V(c) (59+/-10, 69+/-14, 71+/-21 % predicted, respectively) and D(m) (76+/ 22, 53+/-19, 63+/-16 % predicted, respectively) with no differences between groups (p>0.05). Disproportionate reduction of D(m) (D(m)/V(c) % predicted <1) was seen in all diseases (range 0.36-1.89). A mathematical analysis is presented to illustrate that changes in vascular geometry may additionally influence the proportionality of changes in D(m) and V(c). The mathematical analysis suggests that when reduction in patency of some vessels co-exits with compensatory dilatation of the remaining vasculature, a disproportionate reduction in D(m) relative to V(c) may result. CONCLUSIONS: The balance between vascular curtailment and compensatory dilatation may contribute to the variability of the D(m)/V(c) relationship seen in pulmonary vascular disease. Disproportionate reduction in D(m) relative to V(c) may result from this imbalance and need not imply subclinical alveolar membrane and/or parenchymal disease. PMID- 16376537 TI - Clinical and bacteriologic efficacy of telithromycin in patients with bacteremic community-acquired pneumonia. AB - This retrospective analysis was performed to determine the clinical and bacteriologic efficacy of the ketolide antibacterial telithromycin in patients with community-acquired pneumonia (CAP) with pneumococcal bacteremia. Patients 13 years old with radiologically confirmed CAP and a positive blood culture for Streptococcus pneumoniae at screening were analyzed from eight multicenter Phase III/IV clinical trials. In four open-label, non-comparative studies, patients received telithromycin 800 mg once daily for 7-10 days. In four randomized, controlled, double-blind, comparative studies, patients received telithromycin 800 mg once daily for 5-10 days or a comparator antimicrobial (amoxicillin 1000 mg three times daily, clarithromycin 500 mg twice daily, or trovafloxacin 200 mg once daily) for 7-10 days. In total, 118 patients (telithromycin, 94/1061 [8.9%]; comparator, 24/244 [9.8%]) had documented pneumococcal bacteremia. Those who were treated with telithromycin achieved a clinical cure rate of 90.2% (74/82, per protocol population); S. pneumoniae was eradicated in 77/82 (93.9%) bacteremic patients who received telithromycin and 15/19 (78.9%) comparator-treated patients. Clinical cure was also observed among telithromycin-treated bacteremic patients who were infected with penicillin- or erythromycin-resistant strains of S. pneumoniae (5/7 and 8/10, respectively). In conclusion, telithromycin achieves high clinical and bacteriologic cure rates in CAP patients with pneumococcal bacteremia. PMID- 16376538 TI - Comparison of induced sputum inflammatory profiles between childhood and adult onset asthma. AB - The aim of this study was to investigate differences in airway inflammation between childhood and adult-onset asthma. A total of 47 asthmatic subjects were recruited from patients attending outpatient clinic. A group of 32 adults, mean age 42.8 years (yrs) and a group of 15 children, mean age 11.7 yrs were included. The two groups did not differ in respect to gender, dose of inhaled corticosteroids, atopy status or duration of asthma (mean duration 7.75 yr). Lung function tests, and sputum induction were performed. Flowcytometry was used to study cell population and interleukin-8, eosinophilic cationic protein (ECP) and granulocyte-macrophage colony stimulating factor were measured by enzyme-linked immunosorbent assay (ELISA). Three out of 15 (20%) of the children and 6 out of 32 (19%) of the adult patients were unable to produce a sufficient sputum sample. However, all individuals tolerated the procedure well. The viability of induced sputum cells did not differ among adult-onset asthmatics and children with asthma. Children had greater number of total cells in induced sputum compared with adult subjects (P=0.02). No statistical difference in T-lymphocytes subsets was found between the two groups, except for CD25 (P=0.04). A negative correlation was found between forced expiratory volume (FEV1) values and ECP levels (r=0.338, P=0.04) in the whole population (children and adults). Our study showed that the immunopathology of pediatric and adult asthma is similar and sputum induction provides opportunities for comparison of airway inflammation in childhood and adult asthma safely. PMID- 16376539 TI - Natural disasters: a view from afar. PMID- 16376540 TI - Making bio-sense of toxicity: new developments in whole-cell biosensors. AB - Bacterial whole-cell biosensors are very useful for toxicity measurements of various samples. Semi-specific biosensors, containing fusions of stress-regulated promoters and reporter genes, have several advantages over the traditional, general biosensors that are based on constitutively expressed reporter genes. Furthermore, semi-specific biosensors are constantly being refined to lower their sensitivity and, in combination, are able to detect a wide range of toxic agents. However, the requirement for a positive response of these biosensors to toxicants can result in false-negative responses. The application of in situ inoculation and single-cell detection, combined with the introduction of new reporter genes and refined detection equipment, could lead to the extensive use of semi specific, stress-responsive biosensors for toxicity estimations in the future. PMID- 16376541 TI - Key cancer cell signal transduction pathways as therapeutic targets. AB - Growth factor signals are propagated from the cell surface, through the action of transmembrane receptors, to intracellular effectors that control critical functions in human cancer cells, such as differentiation, growth, angiogenesis, and inhibition of cell death and apoptosis. Several kinases are involved in transduction pathways via sequential signalling activation. These kinases include transmembrane receptor kinases (e.g., epidermal growth factor receptor EGFR); or cytoplasmic kinases (e.g., PI3 kinase). In cancer cells, these signalling pathways are often altered and results in a phenotype characterized by uncontrolled growth and increased capability to invade surrounding tissue. Therefore, these crucial transduction molecules represent attractive targets for cancer therapy. This review will summarize current knowledge of key signal transduction pathways, that are altered in cancer cells, as therapeutic targets for novel selective inhibitors. The most advanced targeted agents currently under development interfere with function and expression of several signalling molecules, including the EGFR family; the vascular endothelial growth factor and its receptors; and cytoplasmic kinases such as Ras, PI3K and mTOR. PMID- 16376542 TI - Assessment of hearing in very young children receiving carboplatin for retinoblastoma. AB - Children with retinoblastoma have increasingly been treated with carboplatin in the past decade. Ototoxicity is a known, possible, side-effect of carboplatin. Since retinoblastoma patients are very young and frequently have impaired vision, the evaluation of hearing loss is very important. The hearing status of 25 children with retinoblastoma treated with carboplatin (median cumulative dose 2,240 mg/m(2)) was evaluated in detail. Median age at first carboplatin administration was 7 months. The evaluation of hearing loss was performed by an age-appropriate measurement protocol consisting of tympanometry, otoacoustic emission measurements, auditory brainstem responses and (high-frequency) visual reinforcement audiometry (VRA) or play-audiometry. The median follow-up time after last carboplatin dose was 25 months (range 1-94 months). In none of the children was hearing loss detected after carboplatin administration. A measurement protocol that includes tympanometry, distortion product otoacoustic emission measurements and high-frequency VRA is recommended for young children receiving carboplatin or other ototoxic drugs. PMID- 16376543 TI - Application of dried sewage sludge as phenol biosorbent. AB - The aim of this work was to determine the potential application of dried sewage sludge as a biosorbent for removing phenol from aqueous solution. Results showed that biosorption capacity was strongly influenced by the pH of the aqueous solution with an observed maximum phenol removal at pH around 6-8. Biosorption capacity increased when initial phenol concentration was increased to 110 mg/L but beyond this concentration, biosorption capacity decreased suggesting an inhibitory effect of phenol on biomass activity. Biosorption capacity decreased from 94 to 5 mg/g when biosorbent concentration was increased from 0.5 to 10 g/L suggesting a possible competitive effect of leachable heavy metals from the sludge. The effect of Cu2+ on biosorption capacity was also observed and the results confirmed that the phenol biosorption capacity decreased when concentration of Cu2+ in the sorption medium was increased up to 15 mg/L. Desorption of phenol using distilled deionized water was less than 2% suggesting a strong biosorption by the biomass. PMID- 16376544 TI - The reversed binding of beta-phenethylamine inhibitors of DPP-IV: X-ray structures and properties of novel fragment and elaborated inhibitors. AB - The co-crystal structure of beta-phenethylamine fragment inhibitor 5 bound to DPP IV revealed that the phenyl ring occupied the proline pocket of the enzyme. This finding provided the basis for a general hypothesis of a reverse binding mode for beta-phenethylamine-based DPP-IV inhibitors. Novel inhibitor design concepts that obviate substrate-like structure-activity relationships (SAR) were thereby enabled, and novel, potent inhibitors were discovered. PMID- 16376545 TI - Synthesis and cytotoxicity of new heterocyclic terpenylnaphthoquinones. AB - Several 2-arylamino-, 2-aryloxy- and 2-arylsulfanyl-6(7)-alkyl-1,4 naphthoquinones (NQ) have been prepared and further transformed into the corresponding heterocyclic-fused naphthoquinones by palladium (II)-catalyzed oxidative cyclization. The compounds synthesized have been evaluated against neoplastic cell lines. The extension of the polycyclic system clearly decreased the cytotoxic potency of the 2-substituted terpenylnaphthoquinones. PMID- 16376546 TI - Design and synthesis of novel amino-substituted xanthenones and benzo[b]xanthenones: evaluation of their antiproliferative activity and their ability to overcome multidrug resistance toward MES-SA/D x 5 cells. AB - A number of new xanthenone and benzo[b]xanthenone amino derivatives and their pyrazole-fused counterparts have been designed and synthesized possessing structural analogy to the potent anticancer agent 9-methoxypyrazoloacridine. The synthesis of the compounds proceeds through nucleophilic substitution of 1-chloro 4-nitroxanthenone or the corresponding benzo[b]xanthenone by the appropriately substituted amine or hydrazine, reduction of the nitro group, and conversion into the suitable dialkylaminoacetamides. This method cannot be applied for synthesis of the pyrazole-fused benzo[b]xanthenones, consequently a different, simple, and high-yielding synthetic procedure was developed for the preparation of the target molecules. In vitro cytotoxic potencies of the new derivatives toward the murine leukemia L1210 cell line, human colorectal adenocarcinoma (HT-29), and human uterine sarcoma (MES-SA and its 100-fold resistant to doxorubicin variant MES SA/D x 5) cell lines are described and compared to those of reference drugs. The compounds exhibited significant cytotoxic activity against the tested cell lines and in addition they retain activity against the multidrug resistant MES-SA/D x 5 subline, showing resistant factors close to 1. A number of derivatives were found to possess DNA binding capacity, according to a standard ethidium bromide displacement assay. The majority of the studied compounds induce a G2/M arrest, although among them some G1 or S blockers have also been identified. PMID- 16376547 TI - Synthesis, metal complexation and biological evaluation of a novel semi-rigid bifunctional chelating agent for 99mTc labelling. AB - A novel bifunctional chelating agent bearing an aromatic ring has been synthesised and characterised. This ligand formed well-defined oxorhenium complexes. The analogous 99mTcO-complex was obtained in an excellent yield with high radiochemical purity (>95%). The biodistribution of the 99mTc-complex after intravenous injection studied in normal rats showed that the activity was excreted mainly via renal-urinary pathway indicating its use for labelling peptides with 99mTc. PMID- 16376548 TI - Binding free energy calculations of adenosine deaminase inhibitors. AB - The interactions between four inhibitors and adenosine deaminase (ADA) were examined by calculating their binding free energies after molecular dynamics simulations. A bonded model was used to represent the electrostatic potentials of the zinc coordination site. The charge distribution of the model was derived by using a two-stage electrostatic potential fitting calculations. The calculated binding free energies are in good agreement with the experimental data and the ranking of binding affinities is well reproduced. Notably, our findings suggest that non-polar contributions play an important role for ADA-inhibitor interactions. PMID- 16376549 TI - Synthesis and transdermal permeation-enhancing activity of ketone, amide, and alkane analogs of Transkarbam 12. AB - Transkarbam 12 (5-(dodecyloxycarbonyl)pentylammonium-5 (dodecyloxycarbonyl)pentylcarbamate, T12) is a highly active transdermal permeation enhancer. In this study, ketone, amide, and alkane analogs of T12 have been synthesized and evaluated for their permeation-enhancing activity using porcine skin and theophylline as a model drug. Replacement of ester by methylene and ketone, respectively, led to a significant decrease of activity. Amide analogs displayed lower activity in 60% propylene glycol and were comparable to T12 in isopropyl myristate. An intramolecular H-bond between ester and ammonium carbamate group was suggested to be important for the permeation-enhancing activity of T12. PMID- 16376550 TI - First-principles calculation of the electronic structure and energy loss near edge spectra of chiral carbon nanotubes. AB - We present first principles calculations of the electronic structure of small carbon nanotubes with different chiral angles theta and different diameters (d<1 nm). Results are obtained with a full potential method based on the density functional theory (DFT), with the local density approximation (LDA). We compare the band structure and density of states (DOS) of chiral nanotubes with those of zigzag and armchair tubes with similar diameters. The carbon K-edge energy loss near edge structures (ELNES) have been studied and pi* and sigma* contributions have been evaluated. These contributions give information on the degree of hybridization for the small chiral nanotubes. PMID- 16376551 TI - Improving the energy resolution of X-ray and electron energy-loss spectra. AB - We discuss some practical problems of improving the resolution of X-ray and electron spectra. Iterative Bayesian methods promise greater resolution enhancement than Fourier techniques but they also give rise to spectral artifacts. Satellite peaks are generated adjacent to strong peaks in the original spectrum and oscillatory artifacts become prominent after a large number of iterations, particularly when the original data contain high noise content. In the case of valence-electron energy-loss spectra, satellite peaks are reduced by removing the zero-loss peak prior to spectral sharpening. Even so, care should be exercised in interpreting low intensity at low energy loss (after sharpening) as evidence for a bandgap in the electronic density of states. PMID- 16376552 TI - Algae and UV irradiation: effects on ultrastructure and related metabolic functions. AB - The effects of ultraviolet radiation in the biological relevant wavebands of UV-A (315-400 nm) and UV-B (280-315 nm) on algae have become an important issue as a man-made depletion of the protecting ozone layer has been reported. However, experimental designs to investigate this issue are manifold and the target organisms are extremely diverse. Data are included from the prokaryotic cyanobacteria, haptophytes, diatoms, brown algae to green algae (fresh water, snow algae and marine species) including different habitats from marine littoral and open ocean to freshwater ponds, lakes and snow fields. A broad overview on UV effects on algae is given, with a focus on structurally visible changes. Here we report on destruction in chloroplasts, mitochondria, and the occurrence of structures that are likely to be related to the UV stress. In addition several new data are presented from organisms that have to face naturally high UV irradiation due to their habitats. As no disturbances are reported in these organisms, they obviously have a set of protective mechanisms allowing survival in extreme habitats such as snow fields. Physiological changes as a consequence of UV irradiation are included, effects on the DNA level are summarized, and avoidance strategies are discussed. Every effort has been made to summarize the diverse observations and critically evaluate and compare the different experimental strategies to study UV effects in algae. PMID- 16376553 TI - Analysis of element accumulation in cell wall attached and intracellular particles of snow algae by EELS and ESI. AB - Snow algae frequently occur in alpine and polar permanent snow ecosystems and have developed adaptations to their harsh environment, where extreme temperature regimes high irradiation and low nutrient levels prevail. They live in a unique microhabitat, namely the liquid water between snow crystals. The predominant form appears as 'red snow' and in polar environment also 'green snow' frequently occurs. Light microscopy showed that most cells are densely covered by non-biotic particles of so far unknown composition. As snow normally contains very low amounts of nutrients, introduced mainly airborne like dust and precipitation, the inorganic particles at the surface of the snow algae may be important for their survival. By using electron energy loss spectroscopy (EELS) and electron spectroscopic imaging (ESI), we investigated element distribution in ultrathin sections of snow algae from different polar (Svalbard, 5 m a.s.l., 79 degrees N and maritime Antarctic, King George Island, 10 m a.s.l., 62 degrees S) and alpine habitats (2400-3100 m a.s.l. Tyrol) for the present study. It turned out that the main elements of the cell wall attached particles are Si, Al, Fe and O independently from the origin of the snow algae. Interestingly, the same elements were also found in vacuolar compartments inside the cells. These vacuoles contain electron dense granules or crystals and are frequently found to be connected to the cortical cytoplasm. This finding suggests an uptake mechanism of the respective elements by pinocytosis. Co-transport of toxic aluminium together with silicon may be unavoidable as the inorganic nutrient uptake of the snow algae is limited to the thin water layer between the ice crystals. However, formation of insoluble aluminium silicates may serve as detoxification mechanism. PMID- 16376554 TI - Dermal collagen organization in Bufo ictericus and in Rana catesbeiana integument (Anuran, Amphibian) under the evaluation of laser confocal microscopy. AB - Collagen structural organization plays an important role in the mechanical property of the vertebrate integument. Bufo ictericus and Rana catesbeiana integument was investigated by light microscopy and laser confocal microscopy. Collagenous elements of the dermis were statistical analyzed. The integument is formed by the keratinized squamous stratified epidermis supported by the dermis that is subdivided into the spongious layer with a loose arrangement, and the compact layer formed by collagenous fibers arranged compactly in a criss-crossed manner. Thick collagenous columns have a perpendicular trajectory, and are formed by the assembling of alternating collagenous lamellae in both animals. Short intercolumns of collagenous fibrils connecting collagenous lamellae obliquely or transversally are observed in R. catesbeiana dorsal integument. The present study provides evidences that B. ictericus and R. catesbeiana integument has well organized compact dermis, constituted by collagenous lamellae in a plywood manner. The integument organization is in contrast to the literature in some aspects. This dermal arrangement is important to the biomechanical property of both anuran integuments. PMID- 16376555 TI - Venom gland of Pachycondyla striata worker ants (Hymenoptera: Ponerinae). Ultrastructural characterization. AB - Morphological data concerning the venom gland of worker ants of Pachycondyla striata revealed that this gland consists of three distinct regions: an external secretory portion, composed by a secretory filament that bifurcates in order to give rise to other two filaments; an internal secretory portion, represented by the convoluted gland; and a storage portion, represented by a sac-shaped reservoir. The ultrastructural analysis showed that the reservoir is enveloped by a simple pavementous epithelium, coated internally with a cuticle. The external secretory portion is composed by cells forming a simple cubic epithelium, in which the apical portion presents numerous microvilli while the basal portion of the cells shows infoldings of the plasma membrane containing numerous mitochondria. The convoluted gland possesses cells of irregular morphology with nuclei containing condensed chromatin, suggesting inactivity. However, these cells are in fact undergoing secretory activity, which is probably added to the final secretion produced by the gland. The cytoplasm of these cells contains several elements distributed therein, such as ribosomes and polyribosomes, lipid droplets, and protein inclusions in the form of crystals, thus suggestive of protein storage, which would be used by the insect when metabolically required. PMID- 16376556 TI - Structural and magnetic studies of Co thin films. AB - The trend in reducing device dimension induces new physical properties and requires the development of measurement tools at the nanometer scale. This paper deals with the relation between magnetism and structure of thin films. We have chosen cobalt as a ferromagnetic layer and chromium as a bcc buffer. Magnetic and structural investigations have been led on epitaxial Co/Cr layers grown on MgO (001) substrates. The thickness of the cobalt layer varies from 0.75 to 20 nm. Investigations on the cobalt layer by EXAFS and HRTEM give evidence for a bcc or a hcp structure depending on the cobalt thickness. Magnetic measurements using SQUID indicate that the saturation magnetisation per volume unit is constant for the layers. EELS experiments have been carried out to measure any evolution in the I(L3)/I(L2) ratio for ferromagnetic layers of different thickness. We discuss the influence of structural and magnetic contributions on the evolution of the ratio with the cobalt thickness. PMID- 16376557 TI - Synthesis and electrical properties of ZnO nanowires. AB - Vertically aligned ZnO nanowires were synthesized on the p(+) silicon chip by modifying the CVD process with a vapor trapping design. Scanning electron microscopy was used to investigate the morphology of as-obtained nanowires. X-ray diffraction showed that the obtained nanowires were ZnO crystalline. The rectifying characteristics of the p-n heterojunction composed of ZnO nanowires and a p(+) silicon chip were observed. The positive turn-on voltage was 0.5V and the reverse saturation current was 0.01mA. These vertically aligned ZnO nanowires showed a low field emission threshold of 4V/microm at a current density of 0.1microA/cm(2). The dependence of emission current density on the electric field followed Fowler-Nordheim relationship. PMID- 16376558 TI - The virtual point detector concept for HPGe planar and semi-planar detectors. AB - The virtual point detector assumption was validated for planar and semi-planar detectors. The distance of the virtual point detector from the detector face is larger than the thickness of the detector, which means that the virtual detector position is outside the volume of the detector. PMID- 16376560 TI - Cost-effectiveness analysis of prophylactic respiratory physiotherapy in pulmonary lobectomy. AB - OBJECTIVE: To evaluate the cost-effectiveness balance of implementing an intensive program of chest physiotherapy in pulmonary lobectomy. METHODS: DESIGN: cross-sectional study with historical controls. Cases are 119 patients operated on during a 15-month period of time, after implementation of an intensive chest-physiotherapy program. Controls are 520 patients operated on by the same team before the program started. In these patients, only incentive spirometry was indicated besides routine nursing care. In both series, operative selection criteria and anaesthetic management were similar. Population homogeneity was assessed by comparing age, body mass index (BMI) and estimated postoperative FEV1 (ppoFEV1) of the patients in both series. Selected outcomes were as follows: 30-day mortality, prevalence of respiratory morbidity (atelectasis and pneumonia) and hospital stay. Hospital stay was estimated by Cox regression using age, ppoFEV1, BMI, diagnosis and postoperative morbidity as covariates. Costs were calculated adding chest therapists' salaries and acquisition value of specific training and monitoring devices and its consumable items. Savings from avoided hospitalisation days was discounted. RESULTS: Prevalence of atelectasis and median hospital stay decreased in physiotherapy group. Cost of the program was 48,447.81 (407.12 per treated patient). An estimated total of 151.75 hospital days was saved in the physiotherapy group. Since daily hospitalisation cost is 590.00 in our centre, 89,532.50 savings was estimated from avoided hospitalisation days. CONCLUSIONS: We have found a significant decrease in the rate of postoperative atelectasis without additional costs. In fact, the program has produced considerable monetary savings. PMID- 16376561 TI - Myasthenia gravis developed 30 months after resection of recurrent thymoma. PMID- 16376562 TI - Residual air in the venous cannula increases cerebral embolization at the onset of cardiopulmonary bypass. AB - OBJECTIVE: When the right atrium (RA) cannula is connected to the venous return line of the cardiopulmonary bypass (CPB) circuit, air is often introduced. Air in the venous cannula may increase cerebral air embolization at initiation of CPB despite the arterial line filter. We measured the volume of air present in the venous cannula after cannulation of the RA. Transcranial Doppler quantified emboli as high-intensity transient-signals (HITS) in both middle-cerebral arteries (MCA) at the beginning of CPB. METHODS: After RA cannulation, the air column in the venous line was measured and the total volume calculated using the known lumen diameter. CPB onset was defined as the instant when the CPB machine started moving the patient's blood from the RA into the venous reservoir. Starting from CPB onset, HITS were counted: (a) until completion of the first minute on CPB (1-min count) and (b) until aortic cross clamping (pre-clamping count). RESULTS: We studied 135 patients during coronary artery bypass surgery operated on by 10 cardiac surgeons. HITS during onset of CPB were detected in 95% of patients. Median counts were 10 HITS (25th, 75th percentiles: 3, 26) at 1-min and 21 HITS (8, 51) during pre-clamping. A significant correlation was found between the volume of air in the venous cannula and the HITS counts (r=0.524, p<0.0001). Absence of retained air was associated with lower HITS counts [3 HITS (1, 11)] compared with any amount of air [13 HITS (4, 29), p=0.002)]. The volume of air in the venous cannula, the MCA mean blood flow velocity and the pre clamping time were the only independent predictors of the pre-clamping HITS counts (p<0.001). CONCLUSION: Air in the venous cannula can result in HITS in the MCA. Minimizing the volume of air introduced into the venous cannula after cannulation of the RA can decrease cerebral air embolization at the beginning of CPB. PMID- 16376563 TI - Total arch replacement using a stepwise distal anastomosis for arch aneurysms with distal extension. AB - A total of 120 patients having arch to distal arch aneurysm with downstream extension underwent total arch replacement, with individual arch-vessel reconstruction through median sternotomy using a novel "stepwise" distal aortic anastomosis. Cardiopulmonary bypass was established by cannulating the right axillary artery and the ascending aorta or femoral artery. Hypothermia was at 22 28 degrees C. Through the aneurysm, the descending aorta was divided. Distal anastomosis using the stepwise technique was performed; a tube graft of length 7 12 cm was inserted into the descending aorta and anastomosed by running suture. The distal end of the inserted graft was extracted, and a further four-branched arch graft was joined to it. Selective cerebral perfusion was used for cerebral safety during arch repair. There were three hospital deaths (2.5%). Two patients (1.7%) developed permanent neurological dysfunction and three patients (2.5%) suffered transient cerebral deficits. Three patients (2.5%) required reentry for postoperative bleeding although in none of them bleeding was from the distal anastomosis site with the stepwise technique. Stepwise anastomosis is a useful and secure alternative for distal anastomosis in total arch replacement for arch to distal arch aneurysms with distal extension. PMID- 16376564 TI - Preoperative statin use and in-hospital outcomes following heart surgery in patients with unstable angina. PMID- 16376565 TI - Indexed effective orifice area after mechanical aortic valve replacement does not affect left ventricular mass regression in elderly. AB - OBJECTIVE: After aortic valve replacement, the effects of a small functional prosthesis on the extent and pattern of regression of left ventricular hypertrophy and on clinical outcomes may be less significant in older patients with low cardiac output requirements. The objective of this study was therefore to determine whether patient-prosthesis mismatch affects left ventricular mass regression in the elderly. METHODS: The population studied was made up of 88 patients over 65 years of age with pure aortic stenosis who underwent mechanical aortic valve replacement. The effective orifice area index was calculated for each patient on the basis of the projected prosthesis in vivo effective orifice area. It was considered a continuous variable and influence of its entire range of values on the extent of left ventricular mass regression was analyzed in a multivariate prediction model. RESULTS: Even though, in the group with prosthesis patient mismatch there was a trend for lower postoperative left ventricular mass index (115+/-24 g/m(2) vs 102+/-27 g/m(2), p=0.24) and postoperative peak trans prosthetic gradients (32+/-9.8 mmHg vs 28.9+/-7.79 mmHg, p=0.35) these differences were not statistically significant. The prevalence of residual left ventricular hypertrophy at follow-up was 50% in the group with patient-prosthesis mismatch and 50% in the group without patient-prosthesis mismatch (p=0.83). In multivariate analysis the only factors associated with indexed left ventricular mass were the follow-up time (p=0.015, r(2)=0.22) and preoperative indexed left ventricular mass (p=0.0012, r(2)=0.11). CONCLUSIONS: The major finding of our study is that patient-prosthesis mismatch does not affect left ventricular mass regression in patients older than 65 with pure aortic stenosis who underwent mechanical aortic valve replacement. In older patients with low cardiac output requirements, even a small change in the valve effective orifice area after aortic valve replacement with modern efficient mechanical prosthesis, will result in a marked reduction of pressure gradient and this will be associated with a significant regression of left ventricular mass. PMID- 16376567 TI - Brain oxygenation and metabolism during selective cerebral perfusion in neonates. AB - OBJECTIVE: To investigate the possible neuroprotective effects of selective cerebral perfusion (SCP) during deep hypothermic circulatory arrest on brain oxygenation and metabolism in newborn piglets. METHODS: Newborn piglets 2-4 days of age, anesthetized and mechanically ventilated, were used for the study. The animals were placed on cardiopulmonary bypass, cooled to 18 degrees C and put on SCP (20 ml/(kg min)) for 90 min. After rewarming, the animals were monitored through 2h of recovery. Oxygen pressure in the microvasculature of the cortex was measured by oxygen-dependent quenching of phosphorescence. The extracellular level of dopamine in striatum was measured by microdialysis and hydroxyl radicals by ortho-tyrosine levels. Levels of phosphorylated cAMP response element binding protein (pCREB) in striatal tissue were measured by Western blots using antibodies specific for phosphorylated CREB. The results are presented as mean+/ SD (p<0.05 was significant). RESULTS: Pre-bypass cortical oxygen pressure was 48.9+/-11.3 mmHg and during the first 5 min of SCP, the peak of the histogram, corrected to 18 degrees C, decreased to 11.2+/-3.8 mmHg (p<0.001) and stayed near that value to the end of bypass. The mean value for the peak of the histograms measured at the end of SCP was 8+/-3 mmHg (p<0.001). SCP completely prevented the deep hypothermic circulatory arrest-dependent increase in extracellular dopamine and hydroxyl radicals. After SCP, there was a statistically significant increase in pCREB immunoreactivity (534+/-60%) compared to the sham-operated group (100+/ 63%, p<0.005). Measurements of total CREB showed that SCP did induce a statistically significant increase in CREB as compared to sham-operated animals (168+/-31%, p<0.05). CONCLUSION: SCP, as compared to DHCA, improved cortical oxygenation and prevented increases in the extracellular dopamine and hydroxyl radicals. The increase in pCREB in the striatum following SCP may contribute to improved cellular recovery after this procedure. PMID- 16376568 TI - The state of the actin cytoskeleton determines its association with gephyrin: role of ena/VASP family members. AB - The role the cytoskeleton plays in generating and/or maintaining gephyrin dependent receptor clusters at inhibitory synapses is poorly understood. Here, the effects of actin cytoskeleton disruption were investigated in eGFP-gephyrin transfected cells and hippocampal neurons. While gephyrin was not associated with microfilaments in transfected cells, it colocalized with G-actin and cytochalasin D-induced F-actin patches. The linker region between the MoeA and MogA homology domains of gephyrin was required for colocalization with F-actin patches and for the binding of gephyrin to ena/VASP, an actin anti-capping factor that, in vitro, caused gephyrin binding to polymerized actin. In hippocampal neurons, treatment with cytochalasin D resulted in the redistribution of the neuronal ena/VASP homologue Mena into actin patches and, at early stages of development, a reduction in the number of gephyrin clusters. Our data suggest that Mena binding to F-actin allows for gephyrin recruitment to the leading edge of uncapped actin filaments. PMID- 16376569 TI - The Synthetic Gene Designer: a flexible web platform to explore sequence manipulation for heterologous expression. AB - "Codon optimization" is a general approach to improving heterologous expression where genes are moved from their native genomes into alternatives that exhibit different patterns of codon usage. However, despite reports of successful manipulations and the existence of stand-alone codon optimization software packages or commercial services that offer to redesign genes, the scientific community lacks any systematic understanding of what exactly it means to optimize codon usage. Thus we present a bona fide web application, the "Synthetic Gene Designer," which contrasts with existing software by providing a centralized, free, and transparent platform for the broader scientific community to develop knowledge about synthetic gene design. Consistent with this goal, our software is associated with a moderated e-forum that promotes discussion of synthetic gene design and offers technical support. In addition, the Synthetic Gene Designer presents enhanced functionality over existing software options: for example, it enables users to work with non-standard genetic codes, with user-defined patterns of codon usage and an expanded range of methods for codon optimization. The Synthetic Gene Designer, together with on-line tutorials and the forum, is available at . PMID- 16376570 TI - Cloning, expression, purification, and properties of a putative plasma membrane hexokinase from Solanum chacoense. AB - A full-length hexokinase cDNA was cloned from Solanum chacoense, a wild relative of the cultivated potato. Analysis of the predicted primary sequence suggested that the protein product, ScHK2, may be targeted to the secretory pathway and inserted in the plant plasma membrane, facing the cytosol. ScHK2 was expressed as a hexahistidine-tagged protein in Escherichia coli. Expression conditions for this construct were optimized using a specific anti-hexokinase polyclonal anti serum raised against a truncated version of ScHK2. The full-length recombinant protein was purified to electrophoretic homogeneity using immobilized metal ion affinity chromatography followed by anion exchange chromatography on Fractogel EMD DEAE-650 (S). The purified enzyme had a specific activity of 5.3 micromol/min/mg protein. Its apparent Kms for glucose (23 microM), mannose (30 microM), fructose (5.2 mM), and ATP (61 microM) were in good agreement with values found in the literature for other plant hexokinases. Hexahistidine-tagged ScHK2 was highly sensitive to pH variations between 7.7 and 8.7. It was inhibited by ADP and insensitive to glucose-6-phosphate. These findings constitute the first kinetic characterization of a homogeneous plant hexokinase preparation. The relevance of ScHK2 kinetic properties is discussed in relation to the regulation of hexose metabolism in plants. PMID- 16376571 TI - Molecular cloning and characterisation of prophenoloxidase cDNA from haemocytes of the giant freshwater prawn, Macrobrachium rosenbergii, and its transcription in relation with the moult stage. AB - Expression of prophenoloxidase (proPO) cDNA was determined from haemocytes of the giant freshwater prawn Macrobrachium rosenbergii by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA using oligonucleotide primers based on the proPO sequence of tiger shrimp Penaeus monodon, freshwater crayfish Pacifastacus leniusculus, green tiger shrimp Penaeus semisulcatus, kuruma shrimp Marsupenaeus japonicus, and white shrimp Litopenaeus vannamei. The proPO of M. rosenbergii was constitutively expressed. The 2,547-bp cDNA contained an open reading frame (ORF) of 2,013 bp, a 96-bp 5'-untranslated region, and a 438-bp 3'-untranslated region containing the poly A tail. The molecular mass of the deduced amino acid (aa) sequence (671 aa) was 76.7 kDa with an estimated pI of 7.05. It contained putative copper-binding sites, a complement like motif (GCGWPRHM), a proteolytic activation site, and a conserved C-terminal region common to all known proPOs. However, no signal peptide sequence was detected in giant freshwater prawn proPO. Comparison of amino acid sequences showed that prawn proPO is similar to the proPO of penaeid, crayfish and lobster. Prawn proPO was only synthesised in haemocytes. The proPO transcript was significantly increased in the A stage and achieved the highest level in the B stage, and then declined sharply in the C stage and reached the lowest level in the D(2)/D(3) stage. PMID- 16376572 TI - Different sensitivity of carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) to the immunomodulatory effects of UVB irradiation. AB - In order to study the sensitivity of two fish species, carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss), to the immunomodulatory effects of ultraviolet B (UVB) radiation, the fish were exposed to a single UVB dose of 50, 250, 500 or 1,000 mJ cm(-2). These species represent different phylogenetic groups of fish, and they differ also in their behaviour inhabitating often dark and turbid (carp) or clear and transparent waters (salmonids). Immune responses were studied on day 1 post-irradiation. Unexposed fish, and fish exposed to radiation depleted of UV wavelengths served as controls. UVB irradiation markedly enhanced the blood respiratory burst and cytotoxic activity in carp, but in the head kidney these parameters were significantly suppressed. Rainbow trout respiratory burst was affected only after exposure with the highest dose of UVB. Lymphopenia and granulophilia were noted in both fish blood after exposure. This study indicates that UVB irradiation modulates immune functions in both fish species studied, and that rainbow trout is more tolerant than carp against UVB. Fish are clearly adapted to the environmental UVB levels prevailing in their usual living habitats, but are also a target of undesired effects of UVB on immune functions whenever exposed to increased radiation levels. PMID- 16376573 TI - Change blindness and priming: When it does and does not occur. AB - In a series of three experiments, we explored the nature of implicit representations in change blindness (CB). Using 3 x 3 letter arrays, we asked subjects (Ss) to locate changes in paired arrays separated by 80 ms ISIs, in which one, two or three letters of a row in the second array changed. In one testing version, a tone followed the second array, signaling a row for partial report (PR). In the other version, no PR was required. After Ss reported whether a change had been detected and the PR had been completed (if required), they were asked to identify a degraded letter trigram that was either novel, or from a previously shown row (repetition priming). Our findings indicate that when CB occurs, both the pre-change and post-change stimulus information primes despite its unavailability to consciousness. Surprisingly, findings also indicate that when change detection occurs only the post-change information primes. PMID- 16376574 TI - Learning under anesthesia: checking the light in the fridge? Commentary on Deeprose and Andrade (2006). AB - Research on learning under anesthesia has focused on showing that learning is possible in the absence of awareness. However, a simple dissociation between learning and awareness is conclusive only under strong additional assumptions, and the actual state of consciousness of an anesthetized person is difficult to determine. Instead of trying to establish complete unconsciousness, one might employ gradual anesthesia to experimentally vary the level of consciousness in a controlled fashion, checking whether cognitive processes exist that can change in opposite direction to measures of awareness. PMID- 16376575 TI - Modulation of spectral power and of phase resetting of EEG contributes differentially to the generation of auditory event-related potentials. AB - Nowadays, the mechanisms involved in the genesis of event-related potentials (ERPs) are a matter of debate among neuroscientists. Specifically, the debate lies in whether ERPs arise due to the contribution of a fixed-polarity and fixed latency superimposed neuronal activity to background electroencephalographic oscillations (evoked model) and/or due to a partial phase synchronization of the ongoing EEG (oscillatory model). The participation of the two mechanisms can be explored by the spectral power modulation and phase coherence of scalp EEG rhythms, respectively. However, an important limitation underlies their measurement: the fact that an added neural activity will be relatively phase locked to stimulus, thus enhancing both spectral power and phase synchrony measures and making the contribution of each mechanism less clear-cut. This would not be relevant in the case that an increase in phase concentration was not accompanied by any concurrent spectral power modulation, thus opening the way to an oscillatory-based explanation. We computed event-related spectral power modulations and phase coherence to an auditory repeated-stimulus presentation paradigm with tone intensity far from threshold (90 dB SPL), in which N1 decreases its amplitude (N1 gating) as an attenuation brain process. Our data indicate that evoked and oscillatory activity could contribute together to the non-attenuated N1, while N1 to repeated stimuli could be explained by partial phase concentration of scalp EEG activity without concurrent power increase. Therefore, our results show that both increased spectral power and partial phase resetting contribute differentially to different ERPs. Moreover, they show that certain ERPs could arise through reorganization of the phase of ongoing scalp EEG activity only. PMID- 16376576 TI - Peripheral blood pressure changes induced by dobutamine do not alter BOLD signals in the human brain. AB - In extending the use of functional MRI to neuropharmacology, a primary area of concern is that peripheral blood pressure changes induced by pharmacological agents could independently produce a change in the blood oxygenation level dependent (BOLD) signal, resulting in difficulties distinguishing or interpreting drug-induced neural activations. In the present study, we utilized intravenous dobutamine, a beta-adrenergic receptor agonist, to increase the mean arterial blood pressure (MABP), while examining the effects of MABP changes on the BOLD signal in cocaine-dependent participants. Dobutamine infusion significantly increased the MABP from 93 +/- 8 mm Hg to 106 +/- 12 mm Hg (P < 0.0005), but did not produce a significant global BOLD signal. Yet, a few voxels in the anterior cingulate showed BOLD signal changes that paralleled the changes in blood pressure (BP). Our observations support the conclusion that following the infusion of psychoactive agents, brain BOLD signals accurately reflect neuronal activity, even in the face of relatively large peripheral cardiovascular effects that transiently increase systemic BP. PMID- 16376577 TI - The NIH MRI study of normal brain development. AB - MRI is increasingly used to study normal and abnormal brain development, but we lack a clear understanding of "normal". Previous studies have been limited by small samples, narrow age ranges and few behavioral measures. This multi-center project conducted epidemiologically based recruitment of a large, demographically balanced sample across a wide age range, using strict exclusion factors and comprehensive clinical/behavioral measures. A mixed cross-sectional and longitudinal design was used to create a MRI/clinical/behavioral database from approximately 500 children aged 7 days to 18 years to be shared with researchers and the clinical medicine community. Using a uniform acquisition protocol, data were collected at six Pediatric Study Centers and consolidated at a Data Coordinating Center. All data were transferred via a web-network into a MYSQL database that allowed (i) secure data transfer, (ii) automated MRI segmentation, (iii) correlation of neuroanatomical and clinical/behavioral variables as 3D statistical maps and (iv) remote interrogation and 3D viewing of database content. A population-based epidemiologic sampling strategy minimizes bias and enhances generalizability of the results. Target accrual tables reflect the demographics of the U.S. population (2000 Census data). Enrolled subjects underwent a standardized protocol to characterize neurobehavioral and pubertal status. All subjects underwent multi-spectral structural MRI. In a subset, we acquired T1/T2 relaxometry, diffusion tensor imaging, single-voxel proton spectroscopy and spectroscopic imaging. In the first of three cycles, successful structural MRI data were acquired in 392 subjects aged 4:6-18:3 years and in 72 subjects aged 7 days to 4:6 years. We describe the methodologies of MRI data acquisition and analysis, using illustrative results. This database will provide a basis for characterizing healthy brain maturation in relationship to behavior and serve as a source of control data for studies of childhood disorders. All data described here will be available to the scientific community from July, 2006. PMID- 16376578 TI - Phylogenetic analysis with multiple markers indicates repeated loss of the adult medusa stage in Campanulariidae (Hydrozoa, Cnidaria). AB - The Campanulariidae is a group of leptomedusan hydroids (Hydrozoa, Cnidaria) that exhibit a diverse array of life cycles ranging from species with a free medusa stage to those with a reduced or absent medusa stage. Perhaps the best-known member of the taxon is Obelia which is often used as a textbook model of hydrozoan life history. However, Obelia medusae have several unique features leading to a hypothesis that Obelia arose, in a saltational fashion, from an ancestor that lacked a medusa, possibly representing an example of a rare evolutionary reversal. To address the evolution of adult sexual stages in Campanulariidae, a molecular phylogenetic approach was employed using two nuclear (18S rDNA and calmodulin) and two mitochondrial (16S rDNA and cytochrome c oxidase subunit I) genes. Prior to the main analysis, we conducted a preliminary analysis of leptomedusan taxa which suggests that Campanulariidae as presently considered needs to be redefined. Campanulariid analyses are consistent with morphological understanding in that three major clades are recovered. However, several recognized genera are not monophyletic calling into question some "diagnostic" features. Furthermore, ancestral states were reconstructed using parsimony, and a sensitivity analysis was conducted to investigate possible evolutionary transitions in life-history stages. The results indicate that life cycle transitions have occurred multiple times, and that Obelia might be derived from an ancestor with Clytia-like features. PMID- 16376579 TI - Mutational and functional analyses of xylosyltransferases and their implication in osteoarthritis. AB - OBJECTIVE: The hallmark in osteoarthritis (OA) is the loss of proteoglycans (PGs) in articular cartilage (AC). Xylosyltransferase I (XT-I) catalyzes the transfer of xylose to serine residues in the core protein and initiates the biosynthesis of PGs in AC. The XYLT-II gene encodes a highly homologous protein but its biological function is not yet known. Here we investigate for the first time genetic variations in the XYLT-genes and serum XT-I activities and their implication in OA. METHODS: Denaturing high-performance liquid chromatography (DHPLC) was used for the screening of the XYLT-genes in 49 OA patients. For a detailed characterization of XT-I amino acid exchanges we performed recombinant expression of XT-I mutants in insect cells. Furthermore, the XT activity was measured in the patients' serum. RESULTS: The variation c.1569C>T (XYLT-II) occurs with a significantly higher frequency in younger OA patients in comparison with the older ones (P<0.001) and the controls (P<0.02). Furthermore, significantly higher serum XT activities were found in patients with a long disease duration of OA (P<0.04). The recombinant XT-I mutants p.P385L and p.I552S had reduced enzymatic activity (85% and 74%) compared with the wildtype (wt). CONCLUSIONS: Our findings indicate a correlation of the c.1569 T-allele in XYLT II with an earlier manifestation of OA and that the serum XT activity is a potential biochemical marker for staging and monitoring the progression of AC damage in OA. Comparison of XT-I activity in mutant enzymes in vivo and in vitro revealed that heterozygous mutations are not involved in OA. PMID- 16376580 TI - Electrically induced gel-to-gel phase-transition in neurons. AB - At the end of transcardial perfusions with ice-cold physiological saline for 30 min or with isoosmotic potassium chloride for 5 min, but immediately before perfusion fixation, condenser-discharge electric shocks were administered to rats through surface electrodes pressed onto the temporal muscles of the scalped skull. As a result, striking ultrastructural compaction came about in numerous neurons thinly scattered in certain brain areas. Its features displayed a high degree of similarity to those previously observed following the in vivo administration of the same kind of electric shocks. This surprising independence from the actual state of metabolism questions whether the ultrastructural compaction, induced either in vivo or post mortem, is the result of any cascade of enzyme-mediated processes. On the other hand, a physical mechanism, phase transition propagated by non-covalent free energy stored in a cytoplasmic gel structure, which was proposed recently to explain a mechanically induced similar ultrastructural compaction, appears to apply also to the present case. PMID- 16376581 TI - Role of protein conformation and aggregation in pumping water in and out of a cell. AB - Dialysis cassettes containing BSA solutions were used to simulate passive in vivo conditions to assess the effect of protein conformation and aggregation on cell water content. The cassettes were suspended in dextran solutions to provide a range of fixed osmotic stress values simulating blood plasma. The system was placed on a shaker for 24 h to attain equilibrium. Four manipulation methods; pH, cosolute salt concentration, e.g. NaCl, temperature annealing and urea concentration denaturant were varied to produce well-known manipulations of BSA conformation. It was observed that the cell water content varied from +14% to about -13% with changes in protein conformation and aggregation. The findings demonstrate that a change in protein conformation and aggregation, pumps water in and out of a cell to maintain equilibrium % water content matching the protein conformational hydration parameter. This concept supplements existing theories on cell volume regulation. PMID- 16376582 TI - An NMR method to characterize multiple water compartments on mammalian collagen. AB - A molecular model is proposed to explain water 1H NMR spin-lattice relaxation at different levels of hydration (NMR titration method) on collagen. A fast proton exchange model is used to identify and characterize protein hydration compartments at three distinct Gibbs free energy levels. The NMR titration method reveals a spectrum of water motions with three well-separated peaks in addition to bulk water that can be uniquely characterized by sequential dehydration. Categorical changes in water motion occur at critical hydration levels h (g water/g collagen) defined by integral multiples N = 1, 4 and 24 times the fundamental hydration value of one water bridge per every three amino acid residues as originally proposed by Ramachandran in 1968. Changes occur at (1) the Ramachandran single water bridge between a positive amide and negative carbonyl group at h1 = 0.0658 g/g, (2) the Berendsen single water chain per cleft at h2 = 0.264 g/g, and (3) full monolayer coverage with six water chains per cleft level at h3 = 1.584 g/g. The NMR titration method is verified by comparison of measured NMR relaxation compartments with molecular hydration compartments predicted from models of collagen structure. NMR titration studies of globular proteins using the hydration model may provide unique insight into the critical contributions of hydration to protein folding. PMID- 16376583 TI - Does yeast shmooing mean a commitment to apoptosis? AB - Treatment of yeast Saccharomyces cerevisiae with alpha-pheromone has been reported to lead to massive apoptosis of cells finding no conjugation partner [Severin FF, Hyman AA. Pheromone induces programmed cell death in S. cerevisiae. Curr Biol 2002;12:R233-5]. We report here that this effect is not common in yeast. Using different yeast strains, we demonstrate that identical treatment results in a low mortality even after prolonged treatment with the pheromone. These findings are followed by a general discussion of the biological relevance of apoptosis in yeast. PMID- 16376584 TI - Cyclosporin A does not protect the disruption of the inner mitochondrial membrane potential induced by potassium ionophores in intact K562 cells. AB - Mitochondrial dysfunction has been widely associated with programmed cell death. Studies of intact cells are important for the understanding of the process of cell death and its relation to mitochondrial physiology. Using cytofluorometric approaches we studied the mitochondrial behavior in an erythroleukemic cell line. The effects of protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP), potassium exchanger (nigericin), potassium ionophore (valinomycin), Na+K+-ATPase inhibitor (ouabain) and mitochondrial permeability transition pore inhibitor (cyclosporin A) were evaluated. Cyclosporin A (CSA) was very effective in attenuating the disruption of inner mitochondrial membrane potential induced by CCCP. However, CSA failed to protect the loss of inner mitochondrial membrane potential induced by potassium intracellular flux manipulation. Our findings suggest that mitochondrial cyclophilin is not involved in the cell events mediated by deregulation of potassium flux, underlining the need for further studies in intact tumor cells for a better understanding of the involvement of mitochondria physiology in cell death events. PMID- 16376585 TI - Antiproliferation and apoptosis induced by curcumin in human ovarian cancer cells. AB - Curcumin, an active ingredient from the rhizome of the plant, Curcuma longa, has antioxidant, anti-inflammatory and anti-cancer activities. It has recently been demonstrated that the chemopreventive activities of curcumin might be due to its ability to inhibit cell growth and induce apoptosis. In the present study, we have investigated the effects of curcumin on growth and apoptosis in the human ovarian cancer cell line Ho-8910 by MTT assay, fluorescence microscopy, flow cytometry and Western blotting. Our data revealed that curcumin could significantly inhibit the growth and induce apoptosis in Ho-8910 cells. A decrease in expression of Bcl-2, Bcl-X(L) and pro-caspase-3 was observed after exposure to 40 microM curcumin, while the levels of p53 and Bax were increased in the curcumin-treated cells. These activities may contribute to the anticarcinogenic action of curcumin. PMID- 16376586 TI - Induction of BAALC and down regulation of RAMP3 in astrocytes treated with differentiation inducers. AB - Genes controlling proliferation and differentiation of astrocytes are likely to play an important role in the pathogenesis of astrocytic gliomas and could serve as therapeutic targets. mRNA differential display analysis identified two genes, viz. BAALC and RAMP3, whose expression is altered in primary astrocytes treated with differentiation inducers. BAALC, which has been reported to be expressed in neural progenitor cells, was found to be up regulated in both normal and malignant astrocytes on inhibition of proliferation. RAMP3 functions as a modulator of the receptor for adrenomedullin (AM). AM has been suggested to act as autocrine/paracrine growth factor for gliomas. Our studies show RAMP3 down regulation on staurosporine treatment of astrocytes, suggesting protein kinase C inhibition as a possible strategy for inhibiting AM activity and thereby growth of glioma cells. PMID- 16376587 TI - The human red cell voltage-dependent cation channel. Part III: Distribution homogeneity and pH dependence. AB - The homogeneity of the distribution of the non-selective voltage-dependent cation channel (the NSVDC channel) in the human erythrocyte, and the pH dependence was investigated. Activation of this channel caused a uniform cellular dehydration, which was characterized by the changes in the erythrocyte osmotic resistance profiles: after 1/2 h of activation, the osmolarity at 50% hemolysis changed from 73 mM (control) to 34 mM NaCl, corresponding to 0.48% and 0.21% NaCl respectively. Unchanging standard deviations show participation of the entire erythrocyte population, which implies an even distribution of the NSVDC channel among the cells. Inactivation of the NSVDC channel with N-ethyl-maleimide (NEM) or blocking of the Cl(-) conductance with NS1652 retarded the migration of the resistance profiles towards lower osmolarities. The NSVDC channel activation was blocked by a decrease of the intracellular -- but not the extracellular -- pH. The apparent pK(A) value for the effect was estimated to be 6.5, and the specific histidine reagent 2.4'-dibromoacetophenone (DBAB) inactivated the NSVDC channel. PMID- 16376588 TI - Year ten--and things could not look brighter! PMID- 16376589 TI - Cheyne-Stokes respiration with central sleep apnoea in chronic heart failure: proposals for a diagnostic and therapeutic strategy. AB - Central sleep apnoea (CSA) is highly prevalent in the evolutionary course of chronic heart failure. Such a ventilatory pattern during sleep is independently associated with poor prognosis in people with congestive heart failure. Chronic hyperventilation and daytime hypocapnia are the main mechanisms underlying the frequent association between CSA and cardiac failure. Simplified diagnostic strategies allowing easier recognition of CSA among people with severe heart failure are obviously needed but remain to be validated. Treatment of CSA is essentially aimed at improving cardiac function. When CSA persists, after appropriate adjustment of medication and resynchronisation therapy when indicated, specific ventilatory support during sleep should be considered. Continuous positive airway pressure (CPAP), oxygen, adaptive Servo-ventilation (ASV) and non-invasive ventilation have been proposed. Large randomised trials demonstrating survival and time free from heart transplantation are lacking. PMID- 16376590 TI - Distinguishing sleepiness and fatigue: focus on definition and measurement. AB - Sleepiness and fatigue are two interrelated, but distinct phenomena; observed in a number of psychiatric, medical and primary sleep disorders. Despite their different implications in terms of diagnosis and treatment, these two terms are often used interchangeably, or merged under the more general lay term of 'tired'. Sleepiness is multidimensional and has many causes (multidetermined) and distinguished from fatigue by a presumed impairment of the normal arousal mechanism. Despite its ubiquity, no clear consensus exits as yet as to what constitutes sleepiness. Definitions of sleepiness, to date, are at best operational definitions, conceptualized so as to produce specific assessment instruments. As a result, while a number of subjective and objective measurement tools have been developed to measure sleepiness, each only captures a limited aspect of an otherwise heterogeneous entity. Fatigue is an equally complex phenomenon, its nature captured by a number of conceptualizations and definitions. Measures of fatigue have remained subjective, with a 'gold standard' for its measurement remaining elusive. Despite a high prevalence and high degree of morbidity, fatigue has remained a relatively under appreciated symptom, from both a clinical and research point of view. PMID- 16376591 TI - Sleep function and synaptic homeostasis. AB - This paper reviews a novel hypothesis about the functions of slow wave sleep-the synaptic homeostasis hypothesis. According to the hypothesis, plastic processes occurring during wakefulness result in a net increase in synaptic strength in many brain circuits. The role of sleep is to downscale synaptic strength to a baseline level that is energetically sustainable, makes efficient use of gray matter space, and is beneficial for learning and memory. Thus, sleep is the price we have to pay for plasticity, and its goal is the homeostatic regulation of the total synaptic weight impinging on neurons. The hypothesis accounts for a large number of experimental facts, makes several specific predictions, and has implications for both sleep and mood disorders. PMID- 16376592 TI - Analysis of the Aspergillus nidulans thaumatin-like cetA gene and evidence for transcriptional repression of pyr4 expression in the cetA-disrupted strain. AB - The asexual spore or conidium plays a critical role in the life cycle of many filamentous fungi. However, the process of conidial germination remains surprisingly obscure. To better understand this process at the molecular level we characterized the Aspergillus nidulans cetA gene which is uniquely transcribed in conidiating cultures and whose transcript is significantly enriched in mature conidia. CetA is a member of a novel family of fungal genes of unknown function with homology to plant thaumatin-like (PR-5) defense proteins. We demonstrate by Northern analysis that cetA is a glucose-repressible gene. Transcriptional repression is dependent on the presence of protein kinase A. Western analysis indicates that the CETA protein is absent from conidia but is highly expressed during the first 6h of germination and is secreted into the medium. Disruption of the cetA gene seemingly results in delayed germination, slow growth, abnormal hyphal branching, and cell-wall defects. However, further analysis shows that the mutant phenotype is the result of glucose-dependent transcriptional repression of the pyr4 selectable marker used to disrupt the cetA gene. This is the first time that repression of a selectable marker ("position effect") has been reported in A. nidulans, a finding that may well be of significance in the analysis and interpretation of mutant phenotypes in this organism. PMID- 16376593 TI - Blood cell NO synthesis in response to exercise. AB - Nitric oxide (NO) is important for the maintenance of cardiovascular homeostasis and is also involved in immunity and inflammation. The aim of our work was to determine the effects of intense exercise on plasma and blood cell NO handling. Nine voluntary male professional cyclists participated in the study. Blood samples were taken in basal conditions and 3h after finishing a mountain cycling stage. Exercise-induced neutrophilia, lymphopenia, and hemolysis. Plasma and erythrocytes maintained basal nitrite levels, whereas neutrophils and lymphocytes decreased nitrite concentration after intense exercise. Basal iNOS levels and SOD activity were similar in neutrophils and lymphocytes. iNOS levels and SOD activity dropped in neutrophils and rose in lymphocytes after exercise. Arginase activity rose only in lymphocytes. Neutrophil nitrite was correlated with SOD activity and iNOS levels, but not in lymphocytes. iNOS levels were correlated with SOD in both neutrophils and lymphocytes. Intense exercise maintained plasma basal arginine and ornithine concentration, and decreased citrulline concentration. Intense exercise induced important changes in NO handling in neutrophils and lymphocytes, yet the basal picture was maintained in erythrocytes. PMID- 16376594 TI - The role of glutamate in central nervous system health and disease--a review. AB - Glutamate is the principal excitatory neurotransmitter in the brain. Knowledge of the glutamatergic synapse has advanced enormously over the last 10 years, primarily through application of cellular electrophysiological and molecular biological techniques to the study of glutamate receptors and transporters. There are three families of ionotropic glutamate receptors with intrinsic cation permeable channels. There are also three groups of metabotropic, G-protein coupled glutamate receptors that can modify neuronal excitability. There are also two glial glutamate transporters and three neuronal transporters in the brain. Endogenous glutamate may contribute to the brain damage occurring acutely after traumatic brain injury as well as having a role in the excitatory imbalance present in epileptic conditions and contributing to the pathophysiology of hepatic encephalopathy in animals. Understanding the role of glutamate in these neurological diseases may highlight treatment potentials of antagonists to glutamatergic transmission. This paper presents a review of the literature of glutamate and its role in neurological function and disease. PMID- 16376595 TI - Multiple target screening method for robust and accurate in silico ligand screening. AB - We developed a new in silico multiple target screening (MTS) method, based on a multi-receptor versus multi-ligand docking affinity matrixes, and examined its robustness against changes in the scoring system. According to this method, compounds in a database are docked to multiple proteins. The compounds among these proteins that are likely bind to the target protein are selected as the members of the candidate-hit compound group. Then, the compounds in the group are sorted into descending order using the docking score: the first (n-th) compound is expected to be the most (n-th) probable hit compound. This method was applied to the analysis of a set of 142 receptors and 142 compounds using a receptor ligand docking program, Sievgene [Y. Fukunishi, Y. Mikami, H. Nakamura, Similarities among receptor pockets and among compounds: analysis and application to in silico ligand screening, J. Mol. Graphics Modelling, 24 (2005) 34-45], and the results demonstrated that this method achieves a high hit ratio compared to uniform sampling. We prepared two new scores: the DeltaG score, designed to reproduce the protein-ligand binding free energy, and the hit-optimized score, designed to maximize the hit ratio of in silico screening. Using the Sievgene docking score, DeltaG score and hit-optimized score, the MTS method is more robust than the multiple active-site correction scoring method [G.P.A. Vigers, J.P. Rizzi, Multiple active site corrections for docking and virtual screening, J. Med. Chem., 47 (2004) 80-89]. PMID- 16376596 TI - [Transfusion incidents reports of grade zero: a national and descriptive analysis upon two years of practical survey]. AB - A grade zero transfusion incident is defined as an inappropriate transfusion of blood component due to one or several failures without immediate clinical or biological consequences for the recipient. Two years after the setting up of the mandatory notification of these incidents, Afssaps haemovigilance unit performed a first descriptive national analysis of the data collected in years 2003 and 2004 at the national level. This analysis was based on one part on computarised e Fit national database and on the other part on investigation results documents and additional surveys set up by the network professionals. From a quantitative point of view, this study reveals differences in notification as well as in the type of analysis from one region to another. Quantitatively, 45% of grade zero transfusion incidents correspond to attribution errors. The site of origin of grade zero incidents is for almost 73% linked to health establishment, clinical unit or hospital blood bank, and for almost 23% linked to blood establishment. Complete analysis has notably shown that 9% of the incidents are due to errors in blood component prescription. This descriptive analysis, which identifies recurrent failure and critical points originating from non-appropriated transfusions, should constitute the starting point of a reflection aiming at optimising and standardising methods of analysis of grade zero transfusion incidents and at elaborating suggestions to better control critical points. PMID- 16376597 TI - [Third prize from the jury of the National Syndicate of medical and professional health journals]. PMID- 16376598 TI - Socioeconomic impact of rheumatoid arthritis in Morocco. AB - OBJECTIVE: To estimate the socioeconomic impact of rheumatoid arthritis (RA) in Morocco. MATERIALS AND METHODS: We identified 100 consecutive patients (88 women and 12 men) with RA receiving follow-up either at a teaching hospital or from office-based physicians. For each patient, we recorded direct costs, indirect costs (productivity losses), and intangible costs (deterioration in the social domain of quality of life). RESULTS: Mean age at symptom onset was 31+/- 13.6 years and mean disease duration was 12.8 +/- 7.8 years. RA-related expenses caused financial difficulties for 90% of patients, resulting in poor treatment compliance (61% of cases) and school absenteeism in the children (19% of cases). Of the 34 patients who had paid jobs at symptom onset, 65% stopped working, 6.9 years on average after the diagnosis. Older age, male gender, and a physically strenuous job were associated with stopping work. Six women (10% of married patients) divorced because of their disease. Sexual problems were reported by 67% of patients. The ability to perform domestic chores was affected in 84% of cases and participation in leisure activities in 46% of cases. CONCLUSION: RA has a major socioeconomic impact on affected families. In addition to the disease itself, the low socioeconomic status of many patients and the inadequate social welfare and health insurance systems contribute to the burden. PMID- 16376599 TI - Back problems in Parkinson's disease: an underestimated problem. AB - STUDY DESIGN: Cross-sectional survey. OBJECTIVES: To estimate the extent of back pain in Parkinson's disease (PD). SUMMARY OF BACKGROUND DATA: PD is a common and disabling condition during the course of which back pain may develop. In contrast, the literature on the epidemiology of back pain in PD is poor. METHODS: Patients with PD, seen consecutively in a neurology clinic over a period of 4 months, were inquired about back pain through a self-questionnaire and compared to an age- and sex-matched control group of chronically ill patients. RESULTS: The study involved 104 parkinsonians (mean age: 67.3 years) who had had PD for an average of 11.6 years, and 100 controls (mean age: 65.8 years) who had chronic heart disease or diabetes for an average of 14.2 years. Sixty-two parkinsonians and 23 controls reported back pain. The prevalence was 59.6% in the parkinsonian group and 23.0% in the control group (P < 0.0001). Pain severity was evaluated with a visual analogic scale and averaged 54 +/- 23 mm in parkinsonians and 41 +/ 19 mm in control (P < 0.0001). CONCLUSIONS: Chronic back pain is quite common in PD. It is responsible for a substantial functional impact and needs more attention to reduce disability of such patients. PMID- 16376600 TI - Effects of atrial contraction, atrioventricular interaction and heart valve dynamics on human cardiovascular system response. AB - Various simulation models of different complexity have been proposed to model the dynamic response of the human cardiovascular system. In a related paper we proposed an improved numerical model to study the dynamic response of the cardiovascular system, and the pressures, volumes and flow-rates in the four chambers of the heart, which included the effects of atrial contraction, atrioventricular interaction, and heart valve dynamics. This paper investigates the effects of each one of these aspects of the model on the overall dynamic system response. The dynamic response is studied under different situations, with and without including the effect of various features of the model, and these situations are studied and compared among themselves and to detailed aspects of expected healthy-system response. As an important contribution with potential clinical applications, this paper examines the corresponding effects of atrioventricular interaction, and heart valve opening and closing dynamics to the general system dynamic response. This isolation of physical cause-effect relationships is difficult to study with purely experimental methods. The simulation results agree well with results in the open literature. Comparison shows that introduction of these new features greatly improves the simulation accuracy of the effects of a, v and c waves, and in predicting regurgitant valve flow, the dichrotic notch, and E/A velocity ratio. PMID- 16376601 TI - Bone SPECT reduces the number of unnecessary mandibular resections in patients with squamous cell carcinoma. AB - High percentages (35-78%) of resected mandibles without bone invasion have been reported in squamous cell carcinoma (SCC), adjacent or fixed to the mandible, stressing the need for accurate imaging methods. The aim of this study is to determine the value of bone single photon emission computed tomography (SPECT) for the evaluation of mandibular invasion by SCC. Seventy-nine patients with SCC, adjacent or fixed to the mandible, were included. The results of bone SPECT were compared to the histology of the resection specimens. Bone SPECT was positive in all 50 patients with mandibular invasion and negative in 17 of 29 patients without mandibular invasion. In conclusion, a negative bone SPECT rules out mandibular invasion. Important clinical implications are then that further imaging is not necessary and that the resection can be kept limited. Inclusion of SPECT in the preoperative assessment of these patients will lead to a considerable reduction of unnecessary mandibular resections. PMID- 16376602 TI - Modeling plant morphogenesis. AB - Applications of computational techniques to developmental plant biology include the processing of experimental data and the construction of simulation models. Substantial progress has been made in these areas over the past few years. Complex image-processing techniques are used to integrate sequences of two dimensional images into three-dimensional descriptions of development over time and to extract useful quantitative traits. Large amounts of data are integrated into empirical models of developing plant organs and entire plants. Mechanistic models link molecular-level phenomena with the resulting phenotypes. Several models shed light on the possible properties of active auxin transport and its role in plant morphogenesis. PMID- 16376603 TI - Taeniasis/cysticercosis in Papua (Irian Jaya), Indonesia. AB - Reports showed that an important parasitic zoonotic disease caused by Taenia solium, Taenia saginata and Taenia asiatica is found endemic in several areas of Indonesia including Papua, Bali and North Sumatra. At present it is known that the highest prevalence of taeniasis/cysticercosis in Indonesia, caused by T. solium is among the indigenous communities in Papua (formerly Irian Jaya). In the early 1970s, 8-9% of stool samples from the Enarotali hospital, Paniai District (Irian Jaya) were found positive with Taenia eggs. The samples were from members of the Ekari (Kapauku) ethnic group. Stool samples from the Moni ethnic group, living east of surrounding lakes, were egg negative. Cysticerci of T. solium were discovered in pigs. During the years 1973-1976 cases of burns increased and were ultimately found to be primarily associated with epileptic seizures induced by neurocysticercosis cases. Among 257 cases of burns, 88 cases (62.8%) were suffering from epileptic seizures before or during hospitalization. In the year 1981 T. solium seropositive persons were mostly (16%) found in the endemic Obano village. In 1997 the parasite was discovered in Jayawijaya District, which is located approximately 250 km east of Paniai District. During 1991-1995, a local health center in Assologaima, Jayawijaya District reported 1120 new cases with burns and a further 293 new cases of epileptic seizures among 15,939 inhabitants. The histopathologic appearance and mitochondrial DNA analysis found the cysts to be similar to those of T. solium from other regions of the world. Sensitive and specific serological diagnostic methods were used and improved. Cysticerci were detected in dogs, as well as in pigs. A coproantigen test for detection of adult tapeworms in patients was carried out. Medical treatment with praziquantel for taeniasis and albendazole for cysticercosis (with prednisone and sodium phenytoin treatment in cases with neurocysticercosis) was undertaken. Lifestyle, religion, and socioeconomic aspects are important issues in the perpetuation and enhancing the endemicity of T. solium taeniasis and cysticercosis in Papua, Indonesia. PMID- 16376604 TI - Cystic echinococcosis in the Americas. AB - Echinococcus granulosus and related genotypic variants, the agents of cystic hydatid disease, occur widely in the American continents from Alaska and Northern Canada in North America to Tierra del Fuego in South America. Here we review the historical and current distribution and prevalence of these infections throughout the American countries and the results of programs to control or eliminate the infection. PMID- 16376605 TI - Quantitative analysis of surface EMG activity of cranial and leg muscles across sleep stages in human. AB - OBJECTIVE: The aim of this study was to make a quantitative analysis of the changes in cranial and limb muscle activity from wakefulness to light and deep sleep stages and during rapid eye movement (REM) sleep of normal subjects. METHODS: Polysomnographic recordings were made of the sleep of 9 healthy human subjects, including electromyograms of the suprahyoid, temporalis and masseter cranial muscles and the anterior tibialis limb muscle. Quantitative assessments of EMG activity were carried out with root mean square (RMS) and frequency spectral analysis (FSA) methods. RESULTS: From wakefulness to sleep, a significant reduction (-25.2 to -71.2%; P < 0.01) was observed in EMG activity (for both RMS and FSA) of the 3 cranial muscles using both methods of analysis. The EMG activity of suprahyoid muscle further decreased from non-REM to REM sleep (-17.8 to -43.0%; P < 0.01). In contrast, the EMG activity of the anterior tibialis muscle was only slightly reduced across sleep stages and did not further reduce during REM sleep. During REM sleep, all the 4 muscles maintained minimal activity. CONCLUSIONS: The maintenance of muscle activity during REM sleep suggests that a minimal level of activity is required to preserve physiological functions (e.g. airway patency, posture) related to homeostasis and bodily protection. SIGNIFICANCE: This study suggests that quantitative sleep EMG analysis is important for understanding the mechanisms of sleep-related movement disorders or when objective assessment of changes in EMG activity are needed for diagnostic purposes or for the assessment of drug efficiency. PMID- 16376606 TI - Automated neonatal seizure detection: a multistage classification system through feature selection based on relevance and redundancy analysis. AB - OBJECTIVE: Automatic seizure detection obtains valuable information concerning duration and timing of seizures. Commonly used methods for EEG seizure detection in adults are inadequate for the same task in neonates because they lack the specific age-dependant characteristics of normal and pathological EEG. This paper presents an automatic seizure detection system for newborn with focus on feature selection via relevance and redundancy analysis. METHODS: Two linear correlation based feature selection methods and the ReliefF method were applied to parameterized EEG data acquired from six neonates aged between 39 and 42 weeks. To evaluate the effectiveness of these methods, features extracted from seizure and non-seizure segments were ranked by these methods. The optimized ranked feature subsets were fed into a backpropagation neural network for classifying. Its performance was used as indicator for the feature selection effectiveness. RESULTS: Results showed an average seizure detection rate of 91%, an average non seizure detection rate of 95%, an average false rejection rate of 95% and an overall average detection rate of 93% with a false seizure detection rate of 1.17/h. CONCLUSIONS: This good performance in detecting newborn ictal activities has been achieved based on an optimized subset of 30 features determined by the ReliefF-based detector, which corresponds to a reduction of the number of features of up to 75%. SIGNIFICANCE: The presented approach takes into account specific characteristics of normal and pathological EEG. Thus, it can improve the accuracy of conventional seizure detection systems in newborn. PMID- 16376607 TI - Slow mu variant. PMID- 16376608 TI - Age and gender effects in EEG coherence: III. Girls with attention deficit/hyperactivity disorder. AB - OBJECTIVE: This study investigated intrahemispheric and interhemispheric EEG coherences as a function of age in girls with different DSM-IV subtypes of Attention-Deficit/Hyperactivity Disorder (AD/HD). It completes a series of 3 studies aimed at clarifying developmental and gender impacts on the coupling between brain regions in this disorder. METHODS: Three groups of 40 children (AD/HD combined type, AD/HD inattentive type, and normal controls) participated. Each group contained 8 females in each of 5, 1-year age bands from 8 to 12 years. EEG was recorded from 21 sites during an eyes-closed resting condition. Wave shape coherence was calculated for 8 intrahemispheric electrode pairs (4 in each hemisphere), and 8 interhemispheric electrode pairs, within each of the delta, theta, alpha and beta bands. RESULTS: Developmental effects in intrahemispheric coherences at shorter and longer inter-electrode distances were not as predicted by Thatcher's two-compartment model, contrary to previous findings in boys. Females with AD/HD showed evidence of developmental delay and widespread anomalous elevations in coherence. Girls with AD/HD of the combined type showed similar but greater anomalies than girls with AD/HD of the inattentive type. CONCLUSIONS: Girls with AD/HD show coherence anomalies relative to age- and gender-matched controls, which differ substantially from those shown by boys with AD/HD. These coherence anomalies did not differ in nature between girls with different DSM-IV subtypes of AD/HD, suggesting that subtype differences in girls reflect only symptom severity. SIGNIFICANCE: The data reported here indicate qualitative differences in EEG coherences in girls with AD/HD compared with controls, and quantitative differences between girls with different subtypes of AD/HD. Both sets of effects differ in nature from those previously found in boys with AD/HD, showing the need to carefully consider gender in future studies of AD/HD. PMID- 16376609 TI - EMG findings of facial muscles in ALS. PMID- 16376610 TI - Desmin accumulation restrictive cardiomyopathy and atrioventricular block associated with desmin gene defects. AB - BACKGROUND: Primary desminopathies are caused by desmin gene [DES (MIM*125660)] mutations. The clinical spectrum includes pure myopathies, cardiomuscular diseases and cardiomyopathies. Patients with restrictive cardiomyopathy (RCM) plus atrioventricular block (AVB) due to DES defects are frequently unrecognized unless desmin accumulation is specifically investigated in endomyocardial biopsy (EMB) by ultrastructural study. AIMS: To describe a cardiological phenotype characterized by RCM plus AVB due to desmin accumulation caused by DES defects. METHODS AND RESULTS: Desmin accumulation was diagnosed by means of ultrastructural and immunocytochemical studies of EMB in four unrelated probands with RCM and AVB. Candidate genes [DES and alphaB-crystallin (CRYAB)] were screened using sequence analysis. Four DES gene mutations were identified: three new (R16C, T453I and a 10 bp deletion at the exon-intron boundary of exon 3 disrupting the donor splice site) and one known (R406W). The disease was autosomal dominant in two families, recessive in one and associated with a de novo mutation in one. The mutations cosegregated with phenotype in all patients. CRYAB gene screening was negative. CONCLUSIONS: A cardiac phenotype characterized by RCM and AVB caused by desmin accumulation is associated with DES mutations. Although the mutations affected different domains, the cardiac phenotype was identical. PMID- 16376611 TI - Effect of beta-blocker therapy on functional status in patients with heart failure--a meta-analysis. AB - BACKGROUND: The results of randomised control trials (RCTs) evaluating the effect of beta-blockers on functional status in patients with chronic heart failure are conflicting. AIM: To perform a systematic review and meta-analysis of RCTs evaluating the effect of beta-blockers on New York Heart Association (NYHA) classification and exercise tolerance in chronic heart failure. METHODS AND RESULTS: We selected 28 RCTs evaluating beta-blocker versus placebo in addition to ACE inhibitor therapy. Combined results of 23 RCTs showed that beta-blockers improved NYHA class by at least one class with odds ratio (OR) 1.80 (1.33-2.43) p<0.0001. Meta-analysis of 10 RCTs showed a significant prolongation of exercise time by 44.19 (6.62-81.75) s p=0.021. Combining 8 RCTs evaluating the maximal peak oxygen uptake and 9 RCTs evaluating 6-min walk distance showed that beta blockers had no significant effect compared with placebo, p=0.484, and p=0.730, respectively. Combined results of the 23 RCTs showed significant reducing effect on all cause mortality with OR=0.69 (0.59-0.82) p<0.0001. CONCLUSION: Chronic use of a beta-blocker in conjunction with ACE inhibitor therapy improves dyspnoea and prolongs exercise tolerance time, but has no significant effect on 6-min walk test or maximal oxygen uptake in patients with heart failure. PMID- 16376612 TI - Chronic infarct-related artery occlusion is associated with a reduction in capillary density. Effects on infarct healing. AB - AIM: To assess the relationship between infarct-related artery (IRA) stenosis and capillary density and to assess its effect on scar formation in the human heart. MATERIALS AND METHODS: Morphometric evaluation was performed in 51 human hearts, as follows. Group I non-cardiac death (control), Group II post-Q-wave myocardial infarction (QMI) death and Group III patients who survived QMI and who underwent aneurysmectomy. Using morphometric parameters, the relationship between left ventricle (LV) mass, infarct size, IRA stenosis, cellular hypertrophy and changes in microcirculation were analyzed within the infarcted area and free LV wall. RESULTS: A significant reduction in capillary density within the infarcted area was noted in group II when compared to the control group (1525.6+/-378.5/mm(2) vs. 2968.7+/-457.3/mm(2); p<0.001). Reduction in capillary density was inversely related to infarct size (r=-0.616; p=0.006) and degree of IRA stenosis (r(S)= 0.512; p=0.03). The most significant reduction in capillary density was observed in patients with total IRA occlusion (1204.6+/-156.9/mm(2) vs. 1676.6+/ 245.8/mm(2); p<0.001). Similarly, a reduction in capillary density of over 60% (1030.7+/-241.8/mm(2)) was observed within aneurysms resected surgically. CONCLUSIONS: The study demonstrated precise quantification of the capillary network in patients following QMI. The most significant reduction in capillary density was observed in patients with chronic total IRA occlusion. PMID- 16376613 TI - Lead exposure and birth outcomes in five communities in Shoshone County, Idaho. AB - INTRODUCTION: This study examined birth outcomes in five towns in Shoshone County, Idaho, where residents were exposed to high levels of lead in air emissions during a 6-month period after a fire had damaged the main baghouse (pollution-control device) of a local lead smelter plant in September 1973. METHODS: We studied birth certificate data of 169,878 live singleton infants born to mothers who resided in Idaho at the time of delivery. The outcomes evaluated were preterm infants, small-for-gestational-age (SGA) infants, low birthweight among term infants (TLBW), and mean birthweight among term infants (TMBW). The study compared births in the five towns in Shoshone County (exposed group) to births in the rest of Idaho during three exposure periods: "pre-fire," January 1, 1970-August 31, 1973; "high exposure," September 1, 1973-December 31, 1974; and "post-fire," January 1, 1975-December 31, 1981. RESULTS: During the high-exposure period, the exposed group had an increased prevalence of TLBW (OR=2.4; 90% CI: 1.6-3.6) and SGA (OR=1.9; 90% CI: 1.3-2.8) compared with the rest of Idaho. During the pre- and post-fire periods, the ORs for TLBW were 0.8 and 1.3, respectively, and for SGA, 1.0, and 1.3, respectively. During the high-exposure period, TMBW for the exposed group was 71 g lower than in the comparison group. The TMBW in the exposed group was 8 g lower in the pre-fire period and 26 g lower in the post-fire period than in the comparison group. The study found no increased risk for preterm birth in the exposed group. CONCLUSIONS: Maternal exposures to airborne lead emissions appeared to be associated with increased risks for SGA, TLBW, and reduced TMBW. PMID- 16376615 TI - The carbon starvation-inducible genes Rv2557 and Rv2558 of Mycobacterium tuberculosis are not required for long-term survival under carbon starvation and for virulence in SCID mice. AB - Mycobacterium tuberculosis genes Rv2557 and Rv2558 have no known function. However, proteome, transcriptome and in situ hybridization studies have shown that these genes are significantly upregulated under carbon-starved conditions and in human granulomas, suggesting that they may play a role in persistence. Single and double deletion mutants of M. tuberculosis H37Rv in Rv2557 and/or Rv2558 were generated to explore their individual and/or collective role(s) in growth and survival. The mutants were assessed for growth and survival in vitro under normal and nutrient-deprived conditions and for virulence in the SCID mouse model. Although highly induced by carbon starvation, loss of Rv2557 and/or Rv2558 affected neither the long-term survival of M. tuberculosis under carbon-starved conditions in vitro, nor its virulence in SCID mice. PMID- 16376616 TI - Daily plasma-exchange for life-threatening class I HELLP syndrome with prevalent pulmonary involvement. AB - HELLP syndrome (Hemolysis, Elevated Liver enzymes, Low Platelets) is a thrombotic microangiopathy affecting a minority of women with pre-eclampsia and usually resolves after delivery. The role of plasma exchange in HELLP syndrome has to be defined. Herein is reported a case of a primipara with a class I HELLP syndrome with prevalent pulmonary involvement successfully treated with 8 consecutive plasma exchange plus corticosteroids. In class I HELLP syndrome with cardiopulmonary complications early plasma exchange could be considered a therapeutic option. PMID- 16376617 TI - Oxygen carriers: a selected review. AB - The most common and widely transplanted tissue world wide is blood, which in 2000 resulted in the transfusion of 12.5 million units of blood in the US alone [Goodnough LT, Shander A, Brecher ME. Transfusion medicine: looking to the future. Lancet 2003;361:161-9]. The current use of donated blood products is relatively safe; however, there are inherent problems with allogeneic blood transfusions. The wide spread use of blood in procedures results in problems involving inadequate supply exacerbated in times of war and disasters and by the limited storage life of blood donations (30-42 days). Blood contamination due to patient pre-disposition, poor collection, sterilization, or storage is the second most common cause of death from transfusion in the US [Hillyer CD, Josephson CD, Blajchman MA, Vostal JG, Epstein JS, Goodman JL. Bacterial contamination of blood components: risks, strategies, and regulation: joint ASH and AABB educational session in transfusion medicine. Hematology (Am Soc Hematol Educ Program) 2003:575-89]. Blood is a complex tissue involved in a plethora of homeostatic roles, including immunity, wound healing and the transport of nourishment, electrolytes, hormones, vitamins, heat, oxygen and the removal of metabolic waste products. However, by far the principle role of blood transfusions is the replacement of red cell volume and the maintenance of oxygen levels within the circulation. Creation of investigational new drugs (INDs) which would function as oxygen carriers and prolong shelf life is now a very active arena of scientific research. Several such IND products are now in clinical trials. This article gives an easy to follow concise evaluation of major areas of focus and current testing for each type of blood substitution molecule. PMID- 16376618 TI - Optimization of CD34+ collection for autologous transplantation using the evolution of peripheral blood cell counts after mobilization with chemotherapy and G-CSF. AB - BACKGROUND: Peripheral blood progenitor cells (PBPC) collection after high dose chemotherapy can be influenced by several factors. We searched for parameters that may predict the best day to start harvesting of PBPC in order to collect most CD34+ cells with the least number of aphereses. METHODS: We studied patients who underwent mobilization chemotherapy for autologous transplantation. The influence of age, sex, diagnosis, number of previous chemotherapy cycles, peripheral blood (PB) counts at day of mobilization (D0), day of neutrophils <1.0 x 10(9) l(-1) and day of nadir and interval between both (delta) on harvesting was investigated. Multivariate linear correlation models were built to predict the best harvesting with principles of parsimony. In patients where sequential CD34+ cell count was performed, the theoretical day of peak was calculated by interpolation in polynomial regression. RESULTS: One hundred and thirty four patients entered the analysis: 36 Hodgkin's lymphoma (HL), 65 B-large cell lymphoma (NHL) and 33 multiple myeloma (MM). Day of harvesting correlated with nr CHT, hemoglobin on D0, day of granulocytes <1.0 x 10(9) l(-1), delta and dosis of mobilization therapy. The day of CD34+ peak could be calculated by the formula = (-0.41) x Hemoglobin D0 + (day peripheral CD34+ cells = 10 x 10(6) microl(-1)) x 0.99 + 7.8. This model could explain 81% of the variance of the peak day and was stable by bootstrap resampling. Day of peripheral CD34+ cells = 10 x 10(6) microl(-1) preceded the calculated peak by 3-9 days. CONCLUSIONS: Although the day of best collection can be predicted using only sequential PB counts after mobilization chemotherapy, a model of prediction using peripheral CD34+ cell count is important especially for optimizing collection in poor mobilizing patients. PMID- 16376619 TI - Pharmacogenetics of multigenic disease: heart disease as an example. AB - We now have a greater understanding of the workings of the human genome as well as a wide assortment of pharmacological and mechanical therapies that clearly improve mortality and quality of life for our patients with cardiovascular disease, but these areas of therapeutics and genomics have essentially advanced independently with little interaction up until recently. Pharmacogenetics is the study of the effect of a medication as it relates to single or defined sets of genes. A major goal will be to integrate the two so that true personalized therapy can be delivered. This review explores the clinical implications of the complex interactions in multigenic disease and pharmacology with examples of atherosclerosis and heart failure. The therapies of today are the direct result of understanding the epidemiological, molecular and genetic basis of cardiovascular disease with the application to clinical practice. The complexity of multigenic disease and the promise of pharmacogenetics will require that we improve on the methods of drug evaluation and this includes the need for new statistical methods, bioinformatics, and novel clinical trial design with sufficient power to detect differences in therapy. There must be a continued effort to apply biological and mechanistic plausibility in understanding disease and pharmacology but openness to new ideas and concepts, especially in understanding the workings of the genome. PMID- 16376620 TI - Blood flow and O2 extraction as a function of O2 uptake in muscles composed of different fiber types. AB - We examined how the greater vasodilatory capacity of slow--(ST) versus fast twitch (FT) muscles impacts the relationship between blood flow (Q ) and O2 uptake (VO2) and, consequently, the O2 extraction (a-vO2 diff.)-to-VO2 relationship. Q was measured with radiolabelled microspheres, while VO2 was calculated by the Fick principle using measurements of microvascular O2 pressure (phosphorescence quenching) at rest, low--(2.5 V) and high-intensity contractions (4.5 V) for soleus (Sol; ST, n=5), mixed-gastrocnemius (MG; FT, n=7) and white gastrocnemius (WG; FT, n=7). The slope of the Q-to-VO2 relationship (delta Q/delta VO2] ) was not different among muscles (Sol = 5.5 +/- 0.2, MG = 6.0 +/- 0.11 and WG = 5.8 +/- 0.06; P > 0.05). In contrast, the intercept was greater (P < 0.05) for Sol (16.3 +/- 2.7 ml min(-1) 100 g(-1)) versus MG and WG (in ml min( 1) 100 g(-1): 1.39 +/- 0.26 and 1.45 +/- 0.23, respectively; MG and WG, P > 0.05). In addition, the a-vO2 diff.-to-VO2] relationship for Sol was shifted rightward compared to MG and WG. These data suggest that the increase in Q for a given change in VO2 is similar for slow- and fast-twitch muscles, at least for the range of metabolic rates and muscles studied herein and that a-vO2 diff. differences result from the lower resting Q in FT muscles. PMID- 16376621 TI - Determination of lactate dehydrogenase isoenzymes in single rat glioma cells by capillary electrophoresis with electrochemical detection. AB - A method for determination of lactate dehydrogenase (LDH) isoenzymes in single rat glioma cells (C6) was developed. In this method, a whole cell was electrokinetically injected into the front end of the separation capillary. After that, the cell was lysed by ultrasonication and the isoenzymes in the cell were pre-separated at 20 kV for 5 min and then incubated for 2 min with the enzyme substrates nicotinamide adenine dinucleotide (NAD(+)) and lactate in the capillary electrophoresis running buffer. The electroactive product NADH generated by the isoenzymes through on-capillary enzyme-catalyzed reaction was detected at the outlet of capillary by using the end-capillary amperometric detection with a constant potential mode at a carbon fiber bundle microdisk electrode. Since the amplification of signal via the enzyme reaction, the concentration of nicotinamide adenine dinucleotide (NADH) is much higher than that of LDH. The external standardization was used to quantify isoenzymes in individual cells. Three LDH isoenzymes in single rat glioma cells (C6) were determined and quantified. PMID- 16376622 TI - The current state of newborn health in low income countries and the way forward. AB - Of the 4 million neonatal deaths that occur worldwide each year 99% of these occur in developing countries. South Asia and sub-Saharan Africa regions account for two thirds of the global burden. Skilled professionals attend only 35% deliveries in South Asia and 41% in sub-Saharan Africa. Known, affordable interventions delivered through a rational mix of outreach, family/community and clinical services can reduce over 70% of all neonatal deaths. The Millennium Development Goal of reducing the mortality of children under 5 years by two thirds by the year 2015 from the 1990 baseline would require a substantial reduction in neonatal mortality in the next decade. For this, the low and middle income countries must urgently review their existing programs, and design and implement improved, integrated action plans for maternal, newborn and child health. International community, including the academics, institutions and professional bodies in developed countries can play a crucial role to make this mission a success. PMID- 16376623 TI - Letter to the editor regarding "Bone mineral density of the proximal femur is not related to dynamic joint loading during locomotion in young women." by Bareither et al. PMID- 16376625 TI - Improving the eating patterns of infants and toddlers. PMID- 16376626 TI - Improving the diets of the young: considerations for intervention design. PMID- 16376627 TI - Feeding Infants and Toddlers Study (FITS): development of the FITS survey in comparison to other dietary survey methods. AB - OBJECTIVE: This article describes the steps in the planning and development of the 2002 Feeding Infants and Toddlers Study. METHODS: We describe the study's rationale, sampling methodology, survey questionnaire development, dietary methodology, field data collection, and data processing and analysis. A brief review of existing national nutrition surveys and studies of infants and toddlers, and available study designs and dietary methods, is also included. SUBJECTS/SETTING: Most national studies have been cross-sectional and assessed breastfeeding rates, dietary intake, and nutritional status among general and high-risk populations. Other specialized studies have been longitudinal and tracked dietary intake and nutritional status from infancy to the preschool years, or focused on studying a specific research topic, such as the relationship between fluoride intake and dental caries. CONCLUSIONS: The 2002 Feeding Infants and Toddlers Study has advanced the knowledge base on infant and toddler nutrition by using state-of-the-art methodology and by providing researchers with updated information to develop further research questions. Our findings can be used by child health and nutrition organizations to develop dietary recommendations and improved nutrition education materials. Clinicians and practitioners in the fields of public health and wellness can use the information to provide practical advice to parents in a variety of settings to help give children a more healthful start. PMID- 16376628 TI - Sources of energy and nutrients in the diets of infants and toddlers. AB - OBJECTIVE: To identify major sources of energy and 24 nutrients and dietary constituents in the diets of US infants and toddlers and to describe shifts in major nutrient sources as children age. DESIGN: Data from 24-hour recalls collected in the 2002 Feeding Infants and Toddlers Study were analyzed to determine the percentage contribution of foods and supplements to total intakes of energy, nutrients, and other dietary constituents. A total of 3,586 unique foods and dietary supplements were reported. Reported foods and supplements were classified into 71 groups based on similarities in nutrient content and use. Nine hundred seventy-nine food mixtures were disaggregated into their ingredients and ingredients were classified into one of the 71 groups using the same decision rules that guided classification of foods analyzed at the whole food level. SUBJECTS/SETTING: A national random sample of 3,022 US infants and toddlers 4 to 24 months of age. STATISTICAL ANALYSES PERFORMED: The population proportion formula was used to determine the percentage contribution of each of the 71 groups to total intakes. This was done by summing the weighted amount of a given nutrient provided by a given group for all individuals in the sample and dividing by the total weighted amount of that nutrient consumed by all individuals from all foods and supplements. Groups that provided at least 1% of the nutrient in question were rank-ordered. Separate tabulations were prepared for three age groups (4-5 months, 6-11 months, and 12-24 months). RESULTS: Infant formula, breast milk, and milk are major contributors of energy and most nutrients in the diets of infants and toddlers. Among toddlers, juices and fruit-flavored drinks are the second and third most important sources of energy. Fortified foods make substantial contributions to intakes of many essential nutrients, and these contributions increase as children age. For example, among toddlers, fortified grain-based foods make substantial contributions to intakes of vitamin A, iron, and folate, relative to foods that are naturally rich in these nutrients. Supplements also make substantial contributions to intakes of vitamins and selected minerals, particularly among toddlers. CONCLUSIONS: In assessing dietary intakes of infants and toddlers, dietetics professionals need to carefully consider contributions of fortified foods and supplements. Dietetics professionals should educate caregivers of infants and toddlers about the importance of foods (rather than just nutrients) in promoting health and about the importance of early feeding practices in the development of lifelong eating habits. Caregivers should be encouraged to avoid relying on fortified foods and supplements to meet nutrient needs and educated about the potential risk of excessive intakes. Caregivers of toddlers and infants over 4 to 6 months of age who are consuming solid foods should be encouraged to feed a wide variety of fruits, vegetables, and whole grains, as well as foods naturally rich in iron. PMID- 16376629 TI - Current electrolyte intakes of infants and toddlers. AB - OBJECTIVE: To determine how sodium, chloride, and potassium intakes of today's infants and toddlers compare with the Dietary Reference Intakes (DRIs) of these nutrients established recently by the Food and Nutrition Board of the Institute of Medicine. STUDY DESIGN: Population estimates of usual intake distributions of sodium, chloride (assumed to be equamolar to sodium), and potassium of 4- to 5-, 6- to 11-, and 12- to 24-month-old infants and toddlers were calculated and compared with DRIs of these nutrients by 0- to 6-month-old, 7- to 12-month-old, and 1- to 3-year-old children. SUBJECTS: Infants and toddlers (n=3,022) who participated in the 2002 Feeding Infants and Toddlers Study. STATISTICAL ANALYSES: Means and distributions (percentiles) of the usual intakes of sodium, chloride, and potassium were calculated using Institute of Medicine-recommended procedures and compared with the DRIs (ie, Adequate Intake [AI] and tolerable upper intake level [UL]). RESULTS: Mean sodium and chloride intakes of 4- to 5 month-old infants (188 mg/day and 290 mg/day, respectively) were 57% greater than the AIs (120 mg/day and 180 mg/day) and mean potassium intake (730 mg/day) was 83% higher than the AI (400 mg/day). Mean sodium, chloride, and potassium intakes of 6- to 11-month-old infants were 493 mg/day, 761 mg/day, and 1,225 mg/day, respectively-33%, 33%, and 75% higher than the AIs of these nutrients for this age group (sodium, 370 mg/day; chloride, 570 mg/day; potassium, 700 mg/day). Even the 10th percentile of potassium intake of this age group was greater than the AI. The usual mean sodium and chloride intakes of 12- to 24-month-old toddlers (1,638 mg/day and 2,528 mg/day, respectively) were 64% higher than the AIs (1,000 mg/day and 1,540 mg/day, respectively) and the usual mean sodium and chloride intakes of 58% of this age group were above the ULs. In contrast, mean potassium intake of 12- to 24-month-old toddlers (1,971 mg/day) was only 66% of the AI (3,000 mg/day). At all ages, sources of sodium, chloride, and potassium intakes reflected current feeding guidelines, primarily human milk and formula prior to 6 months of age and primarily cow's milk and table foods after 1 year of age. CONCLUSIONS: Mean sodium and chloride intakes of infants and toddlers who participated in the 2002 Feeding Infants and Toddlers Study exceeded the recently established AIs of these nutrients and the mean intake of 58% of toddlers exceeded the ULs. Mean potassium intake of infants also exceeded the AI of potassium, but the mean potassium intake of toddlers was only 66% of the AI. Whether current intakes of sodium, chloride, and potassium by infants and toddlers are problematical is not clear. Nonetheless, it seems desirable to bring these intakes closer to AIs. This can be accomplished by continuing breast- or formula-feeding and delaying the introduction of cow's milk; limiting the amount of salt added to home-prepared foods; limiting the intake of high-sodium foods, such as processed meats and salty snacks; and increasing the intake of fruits (high potassium and low sodium content) and vegetables (moderate potassium and sodium content). PMID- 16376630 TI - Feeding Infants and Toddlers Study: do vitamin and mineral supplements contribute to nutrient adequacy or excess among US infants and toddlers? AB - OBJECTIVE: To report the prevalence of dietary supplement use in a random sample of US infants 4 to 24 months of age, and to compare demographic characteristics, usual nutrient intakes, and food patterns of supplement users and nonusers. DESIGN: Data from 24-hour recalls collected for the 2002 Feeding Infants and Toddlers Study were analyzed. Recalls included nutrient contributions from dietary supplements as well as all foods and beverages. We estimated usual energy and nutrient intakes of supplement users and nonusers, as well as the prevalence of nutrient adequacy and excess in the two groups. We also compared demographic characteristics and food patterns of supplement users and nonusers and, for supplement users, estimated the proportion of total intake provided by foods and the proportion provided by supplements. SUBJECTS: A national random sample of 3,022 infants and toddlers age 4 to 24 months, including 430 vitamin and/or mineral supplement users and 2,592 nonusers. STATISTICAL ANALYSIS: We compared means, percentile distributions, and proportions by age and supplement subgroup, and applied the Dietary Reference Intakes to assess usual nutrient intakes. We conducted regression analysis to determine which population characteristics predict the use of dietary supplements in this population. RESULTS: Overall, 8% of infants age 4 to 5 months received some type of dietary supplement. The prevalence of supplement use increased with age, to 19% among infants 6 to 11 months and 31% among toddlers 12 to 24 months. The vast majority of supplement users (97%) received only one type of supplement, most commonly a multivitamin and/or mineral supplement. Vitamin/mineral supplement use among infants and toddlers was associated with being a first-born child and being reported by the primary caretaker as being a picky eater. Characteristics that were independent predictors of supplement use were living in the Northeast, being male, and living in a household with fewer children. We found no significant differences between supplement users and nonusers in mean daily intakes of nutrients or nutrient density from foods alone, and few differences in food consumption. Overall, the prevalence of inadequate intakes was low (<1% to 2%). However, 65% of supplement nonusers and 9% of supplement users had vitamin E intakes less than the Estimated Average Requirement. Excessive intakes (ie, intakes above the Tolerable Upper Intake Level) were noted for both supplement users and nonusers for vitamin A (97% and 15% of toddlers) and zinc (60% and 59% of older infants and 68% and 38% of toddlers) as well as for folate among supplement users (18% of toddlers). CONCLUSIONS: Generally, healthy infants and toddlers can achieve recommended levels of intake from food alone. Dietetics professionals should encourage caregivers to use foods rather than supplements as the primary source of nutrients in children's diets. Vitamin and mineral supplements can help infants and toddlers with special nutrient needs or marginal intakes achieve adequate intakes, but care must be taken to ensure that supplements do not lead to excessive intakes. This is especially important for nutrients that are widely used as food fortificants, including vitamin A, zinc, and folate. PMID- 16376631 TI - Average portions of foods commonly eaten by infants and toddlers in the United States. AB - OBJECTIVES: To examine average portion sizes consumed per eating occasion by infants and toddlers. Average portions reported for toddlers were compared to average portions for comparably aged children reported in the 1994 to 1996 and 1998 Continuing Survey of Food Intakes by Individuals. In addition, reported average portions were compared with minimum required portion sizes for meals served to infants and toddlers in the Child and Adult Care Food Program (CACFP). DESIGN: Data from 24-hour recalls collected in the 2002 Feeding Infants and Toddlers Study (FITS) were analyzed. Average portion sizes were determined for major food groups and individual foods that were reported by at least 5% of the population. Most foods were reported separately; however, sandwiches were disaggregated into their components. Gram weights of portions consumed were converted, on a food-by-food basis, to household units so that foods with different volume-to-weight ratios could be analyzed together. SUBJECTS/SETTING: A national random sample of 3,022 US infants and toddlers 4 to 24 months of age. STATISTICAL ANALYSES PERFORMED: For each food and food group, average portion sizes per eating occasion were computed for up to six age groups. An average per eating occasion portion was determined for each child who consumed a given food by summing the total amount of food consumed over the day and dividing by the number of eating occasions. These estimates were then summed across all children who consumed the food and divided by the total number of consumers. The number of eating occasions was defined as the total number of times a child had anything to eat or drink during the day, excluding eating occasions that included only water and/or supplements. RESULTS: For most foods, there was a gradual increase in the average portion as age increased. Average portions reported for FITS toddlers were consistent with those reported for comparably aged children in the most recent Continuing Survey of Food Intakes by Individuals. The average portions reported for FITS infants and toddlers were consistent with CACFP-recommended portion sizes for formula, juice, meats, and cheese. For milk (toddlers only), cereal, breads, fruits, and vegetables, average portions reported in FITS were consistently larger than CACFP portion sizes. Distributions showed that, in many cases, the per-eating occasion portion sizes of 50% to 90% of FITS infants and toddlers exceeded the CACFP portion sizes. CONCLUSIONS: Dietitians, pediatricians, and health educators can use the data presented in this article to provide guidance to parents and caregivers about reasonable portion sizes for infants and toddlers. The data should also be useful to those who plan meals for infants and toddlers in child care settings and to researchers studying dietary intakes of infants and toddlers. Advice about reasonable portion sizes should always be tempered with appropriate cautions about avoiding coercive "clean your plate" feeding practices. Parents and caregivers should be encouraged to offer infants and toddlers appropriate portions of healthful foods from the basic food groups, with a special emphasis on fruits, vegetables, and whole grains, and allow them to eat until they are satiated. PMID- 16376632 TI - Relationship between portion size and energy intake among infants and toddlers: evidence of self-regulation. AB - OBJECTIVES: To assess whether dietary intakes of infants and young toddlers show evidence of energy self-regulation. DESIGN: Data from 24-hour recalls collected in the 2002 Feeding Infants and Toddlers Study were analyzed. Multivariate regressions were used to explore the relationship between portion size and usual energy intake as well as the relationship between portion size, number of eating occasions, number of unique foods, and energy density. SUBJECTS/SETTING: A national random sample of 3,022 US infants and toddlers 4 to 24 months of age. STATISTICAL ANALYSES PERFORMED: To measure variability in portion size, an average portion size z score was computed for each child in the sample, across 45 different food groups. The number of eating occasions was defined as the total number of times a child had anything to eat or drink during the day, excluding eating occasions that included only water and/or supplements. The total number of unique foods in a day was defined as the number of unique food codes included in the 24-hour recall, and energy density was computed as kilocalories/gram, including all foods, beverages, and water. Linear regression models were used to assess the effect of portion size and other self-regulation mechanisms on energy intake and to assess the effect of these self-regulation mechanisms on portion size. Separate analyses were performed for three age groups: 4 to 5 months, 6 to 11 months, and 12 to 24 months. RESULTS: A significant negative association was found for all age groups between the number of eating occasions and average portion size z scores, indicating that children who eat less often during the day consume larger-than-average-portion sizes and children who eat more often during the day consume smaller-than-average portions. For infants (11 months and younger), a significant negative association was noted between energy density and average portion size z scores, indicating that, as the energy density of the diet goes down, infants consume larger-than-average portions and, as the energy density of the diet goes up, they consume smaller-than-average portions. Among infants 6 to 11 months, there was a significant positive relationship between portion size and the number of unique foods consumed. For toddlers, there was no association between average portion size z scores and energy density, suggesting that energy self-regulation mechanisms are diminished in this age group. CONCLUSIONS: Our findings confirm the presence of energy self-regulation among infants and young toddlers. These findings can be used to assure parents and caregivers that infants have an innate ability to regulate energy intake. At the same time, it is important to educate parents and caregivers about the potential for environmental cues to diminish natural hunger-driven eating behaviors, even among young toddlers. Dietetics professionals should emphasize the potential adverse effects that coercive feeding behaviors can have on children's innate ability to regulate energy intake. This includes not only admonitions to "clean your plate," but overrestriction of intake that may be motivated by concerns that children are overeating. PMID- 16376633 TI - Feeding Infants and Toddlers Study: characteristics and usual nutrient intake of Hispanic and non-Hispanic infants and toddlers. AB - OBJECTIVE: To compare demographic and maternal characteristics and usual nutrient intakes of Hispanic and non-Hispanic infants and toddlers 4 to 24 months of age in the United States. DESIGN: We conducted three interviews by telephone to collect information on sociodemographic and maternal characteristics, feeding practices, and dietary intake in the 2002 Feeding Infants and Toddlers Study. We collected 24-hour dietary recalls, including a second day's intake on a subsample, using the Nutrition Data System for Research. We used the Personal Computer version of the Software for Intake Distribution Estimation to estimate usual nutrient intake and nutrient adequacy and excess for three age subgroups infants 4-5 months, infants 6-11 months, and toddlers 12-24 months-and Hispanic or non-Hispanic ethnicity. SUBJECTS: A national sample of 3,022 infants and toddlers age 4-24 months, including 371 Hispanic and 2,637 non-Hispanic subjects. STATISTICAL ANALYSIS: We compared means, percentile distributions, and percentages by age/Hispanic ethnicity subgroup, and applied the Dietary Reference Intakes to assess nutrient intakes. RESULTS: Mothers of Hispanic infants and toddlers were younger, less likely to be married, and had lower education levels than mothers of non-Hispanic infants and toddlers (P<.01). Hispanic infants and toddlers had significantly higher rates of participation in the Special Supplemental Nutrition Program for Women, Infants, and Children than non-Hispanic infants and toddlers (42% to 23%) and were more likely to reside in urban areas and have lower annual household income levels (P<.01). There were no significant differences in usual energy intake between Hispanic and non-Hispanic infants and toddlers, and mean usual energy intake exceeded the mean estimated energy requirement for all age/ethnicity subgroups. Hispanic toddlers consumed a significantly higher proportion of energy from carbohydrate (56% to 53%, P<.01) and a significantly lower percentage of energy from fat (31% to 33%, P<.01) than non-Hispanics. Comparing usual mean intakes, Hispanic infants age 6 to 11 months had a significantly lower intake of calcium than non-Hispanics (means of 574 mg and 626 mg per day, respectively, P<.05) and a significantly higher intake of sodium compared with non-Hispanics of the same age (means of 647 mg to 476 mg per day, P<.01). For infants, mean usual intakes were adequate for all nutrients. For toddlers, the prevalence of nutrient inadequacy was low (<1%) with the exception of vitamin E, which was inadequate for 39% of Hispanic toddlers and 50% of non Hispanic toddlers. For nutrients with defined Tolerable Upper Intake Levels, more than one third to almost half of toddlers exceeded the Tolerable Upper Intake Levels for vitamin A and zinc, and more than half (53% and 58% for Hispanics and non-Hispanic toddlers, respectively) exceeded the Tolerable Upper Intake Level for sodium. Usual mean intakes of vitamins A, C, and E and folate, potassium, and fiber were significantly higher among Hispanic toddlers compared with non Hispanic toddlers. CONCLUSIONS: The Feeding Infants and Toddlers Study data provide information that is useful to practitioners, Special Supplemental Nutrition Program for Women, Infants, and Children program staff, and parents for delivering nutrition education messages that are consistent with dietary guidance for infants and toddlers as well as compatible with cultural preferences. PMID- 16376634 TI - Feeding Infants and Toddlers Study: the types of foods fed to Hispanic infants and toddlers. AB - OBJECTIVE: To assess the prevalence of breastfeeding and formula feeding, the age of introduction to specific foods, and the types of foods and beverages consumed by Hispanic infants and toddlers compared with similarly aged non-Hispanic infants and toddlers living in the United States. DESIGN: Descriptive and comparative analysis of dietary recall data and responses to specific interview questions, which were collected in the 2002 Feeding Infants and Toddlers Study. Breastfeeding status, timing of introduction of complementary foods, percentage consuming foods from specific food groups, and the most frequently consumed fruits and vegetables by Hispanic and non-Hispanic children by age group (4-5 months, 6-11 months, 12-24 months). SUBJECTS: A national random sample of 371 Hispanic and 2,637 non-Hispanic infants and toddlers between the ages of 4 and 24 months. STATISTICAL ANALYSIS: To test for differences between Hispanic and non Hispanic children in the percentage who consumed a particular food item, we calculated percentages and standard errors in SUDAAN and 95% and 99% confidence intervals. The most frequently consumed fruits and vegetables were determined by tallying the percentage of infants and toddlers who consumed each specific fruit or vegetable on a given day. RESULTS: Although there were some similarities, the early flavor and food experiences of Hispanic infants were different from similarly aged non-Hispanic infants in several ways. Hispanic infants younger than 1 year of age were more likely to have ever been breastfed and those who were 4 to 5 months were more likely than non-Hispanics to be eating pureed baby foods on a daily basis. Although less likely to be eating non-infant cereals and baby food vegetables, 6- to 11-month-old Hispanics were more likely to be eating fresh fruits, fruit-flavored drinks, baby cookies, and foods such as soups, rice, and beans that are common in many Hispanic cultures. When fruits were introduced into the Hispanic child's diet, they were most commonly consumed fresh. This higher prevalence of being fed soups, rice, beans, and sweetened fruit-flavored drinks as well as tortillas was also observed among the 12- to 24-month-old toddlers. CONCLUSIONS: Dietetics professionals should be aware of the cultural differences in the foods fed to infants and toddlers that may contribute to the development of long-term food preferences and impact on nutrition. Understanding the factors that underlie food preferences is important if we are to develop evidence-based strategies to improve children's eating habits and lower their risks factors associated with obesity and other chronic diseases. PMID- 16376635 TI - Feeding Infants and Toddlers Study: meal and snack intakes of Hispanic and non Hispanic infants and toddlers. AB - OBJECTIVE: To describe meal and snack patterns of Hispanic and non-Hispanic infants and toddlers. DESIGN: A cross-sectional telephone survey in which mothers or other primary caregivers reported their infants' and toddlers' food and beverage intake for a 24-hour period. SUBJECTS/SETTING: Subjects were a subset of the national random sample of children aged 4-24 months who participated in the 2002 Feeding Infants and Toddlers Study. The Feeding Infants and Toddlers Study includes a stratified random sample of 3,022 infants and toddlers aged 4-24 months. Three hundred seventy-one Hispanic and 2,637 non-Hispanic children who had 24-hour dietary recalls are included in the subset. ANALYSES: Means+/ standard errors of daily intakes of energy, nutrients, and nutrient densities were calculated, as were percentages of children consuming foods at each eating occasion. RESULTS: Hispanic and non-Hispanic infants and toddlers, on average, were fed seven times per day. Overall, the percentages of children who ate snacks increased with age, and more than 80% of toddlers aged 12-24 months consumed afternoon snacks, with more than 90% of Hispanic children consuming an afternoon snack. In each age group, there were significant differences between ethnic groups in nutrient intakes by eating occasion. No significant difference was seen for energy across all meal occasions. At age 6-11 months, Hispanic children had a significantly lower intake of carbohydrate at dinner and lower intake of saturated fat at afternoon snacks compared with non-Hispanic children (P<.05). The main difference between Hispanic children's and non-Hispanic children's intakes by eating occasion is at age 12-24 months. Hispanics aged 12-24 months had significantly (P<.05) lower percentages of energy from fat and saturated fat and a significantly (P<.05) higher percentage of carbohydrate at lunch compared with non-Hispanic children. For dinner, Hispanic toddlers had significantly (P<.05) lower intakes of total fat and saturated fat compared with non-Hispanic toddlers at age 12-24 months. Overall fiber intake contributed 2 g/meal for both ethnic groups. Snacks contributed, on average, less than 1 g fiber, except Hispanic toddlers had significantly higher fiber intake at afternoon snacks (1.5 g) than non-Hispanic toddlers. Foods frequently consumed at meals and snacks were lacking in whole grains, vegetables, and fruits. Most nutrients were not significantly different between Hispanics and non-Hispanics for meals and snacks. CONCLUSIONS: Considering the sizeable contribution that snacks make toward overall energy, parents and caregivers should plan toddlers' snacks to complement meals by including additional fruits, vegetables, and whole grains that are culturally appropriate rather than fruit drinks, cookies, and crackers. This will increase fiber intake and limit fat and sugar intakes. To develop healthful eating patterns, introduce toddlers to foods eight to 10 times to increase food acceptance and the likelihood of establishing healthful eating patterns. Dietetics professionals need to consider cultural differences when developing meal and snack patterns for Hispanic and non-Hispanic infants and toddlers. PMID- 16376636 TI - Nutrient intakes and food patterns of toddlers' lunches and snacks: influence of location. AB - OBJECTIVE: To describe nutrient intake and food patterns of lunches and snacks eaten at various locations by US toddlers. DESIGN: A national, cross-sectional telephone survey in which mothers and primary caretakers reported toddlers' food and beverage intake for a 24-hour period. SUBJECTS: Toddlers (n=632), aged 15 to 24 months, a subset in the 2002 Feeding Infants and Toddlers Study. ANALYSES: Means+/-standard errors of the mean, percentages, t tests of mean differences, mean energy and nutrient intake, and nutrient density of toddlers' lunches and snacks. RESULTS: Overall, on any given day, 42.6% of toddlers consumed all meals and snacks at home, 8.1% consumed any meal or snack at day care (and others at home), and 49.3% consumed any meal or snack away from home (all other locations excluding day care). Mean energy intake at lunch ranged from 281 kcal at home to 308 kcal away from home to 332 kcal at day care. There were no significant differences in mean macronutrient intake or fiber intake across locations, but lunches eaten at day care were significantly higher in calcium, phosphorus, magnesium, vitamin D, potassium, and riboflavin compared with those eaten at home or away (P<.05). Mean trans fat intake was significantly (P<.01) lower for lunches consumed at home compared with away from home. For lunches consumed at away locations, the most frequently consumed item, by 35% of toddlers, was french fries. Carbonated beverages were consumed at away lunches by 16% of toddlers, compared with 3% at home and none at day care. Morning snacks provided 124 to 156 kcal and afternoon snacks provided from 139 to 170 kcal, depending on the location. Foods typically eaten at morning snacks for all locations were water, cow's milk, crackers, and 100% juice. Beverages frequently consumed at afternoon snacks were water, whole cow's milk, fruit-flavored drinks, and 100% apple juice. The most frequently consumed foods for an afternoon snack at home or day care were crackers or non-baby food cookies. CONCLUSIONS: Nutritious choices such as milk, fruits, vegetables, and whole grains need to be encouraged in a variety of forms to give toddlers an opportunity to build broader food preferences for life. The consumption of milk at home and other locations, such as restaurants and friends' homes, needs to be encouraged in place of fruit-flavored drinks or other sweetened beverages. For lunches at home, parents may be especially receptive to suggestions about appropriate and easy-to-serve foods, homemade or commercial, for a toddler's lunches and snacks. Day-care providers should be encouraged to use menu planning aids, such as those available from the US Department of Agriculture, even if they are not regulated by a government program. PMID- 16376637 TI - Maternal and child characteristics associated with infant and toddler feeding practices. AB - OBJECTIVE: To describe maternal/child characteristics associated with important practices of feeding US infants and toddlers aged 4 to 24 months. DESIGN: Cross sectional analysis of data collected in the 2002 Feeding Infants and Toddlers Study. Maternal/child characteristics associated with compliance to American Academy of Pediatrics feeding guidelines, and maternal/child characteristics associated with specific feeding patterns were assessed. SUBJECTS: A national random sample of mothers (n=2,515) whose infants and toddlers aged 4 to 24 months made up the Feeding Infants and Toddlers Study cohort. STATISTICAL ANALYSIS: Student t tests were used to compare the means and standard errors and were considered significant if P<.05. To predict if the mother/child met a particular recommendation, logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals. RESULTS: Having a college education was the maternal characteristic associated with the largest number of positive child feeding behaviors. Mothers with a college education were significantly more likely than mothers without a college education to initiate breastfeeding and breastfeed the child to age 6 and 12 months (OR 2.8, 3.2, and 3.9, respectively). College educated mothers were significantly more likely to comply with the American Academy of Pediatrics juice and complementary feeding recommendations (OR 1.4 and 2.0). In addition, infants and toddlers whose mother had a college education were more likely to consume fruit and less likely to consume sweetened beverages and desserts or candy. Ever breastfeeding the sample child, living in the western region of the United States, and being married and older were also associated with multiple positive practices. The child being in day care was associated with decreased duration of breastfeeding at age 6 and 12 months as well as with consumption of salty snacks. CONCLUSIONS: Initiatives to improve infant and toddler feeding practices should focus on assisting mothers who have less than a college education, who are unmarried, whose child is in day care, or who are enrolled in the Special Supplemental Nutrition Program for Women, Infants, and Children. PMID- 16376638 TI - Visual experiences during vitreous surgery under regional anesthesia: a multicenter study. AB - PURPOSE: To investigate patients' subjective intraoperative visual experiences during vitreous surgery performed under regional anesthesia, to ascertain if patients were frightened by their visual experiences, and to determine the risk factors associated with a frightening visual experience. DESIGN: Multicenter, prospective study. METHODS: Sixty-five patients who had vitreous surgery under regional (retrobulbar or peribulbar) anesthesia in five centers in Ireland, Singapore, and Hong Kong were interviewed within 2 hours of their operation using a standardized questionnaire. RESULTS: Thirty patients (46.2%) perceived light perception throughout the entire operation, 19 patients (29.2%) experienced transient loss of light perception, and 16 patients (24.6%) experienced no light perception throughout the entire duration of the surgery. Nine patients (13.8%) were frightened by their intraoperative visual experiences. Patients who were frightened by their visual experiences were more likely to see color (100%) than those who were not frightened (55.4%) (P = .010). The mean age of the patients who were frightened was lower (51.8 years) compared with those who were not frightened (64.6 years) (P = .003). The mean duration of surgery was longer for patients who were frightened (118.9 minutes) compared with those who were not frightened (91.2 minutes) (P = .047). CONCLUSIONS: Most patients undergoing vitreous surgery under regional anesthesia retained at least light perception intraoperatively. Importantly, 13.8% of patients were frightened by their visual experiences. A younger age, longer duration of surgery, and perception of color were risk factors for a frightening visual experience. PMID- 16376639 TI - Ultrahigh resolution optical coherence tomography in macular dystrophy. AB - PURPOSE: To visualize and investigate intraretinal changes in macular dystrophies with ultrahigh resolution optical coherence tomography (UHR OCT). DESIGN: Prospective observational case series. METHODS: setting: Department of Ophthalmology and Center for Biomedical Engineering and Physics, Christian Doppler Laboratory, Medical University of Vienna, Vienna, Austria. patients: Thirteen patients (23 eyes) with adult-onset foveomacular vitelliform dystrophy (AOFVD) and 14 patients (27 eyes) with Stargardt's disease (SD) or fundus flavimaculatus (FF). OBSERVATIONS: Imaging using a compact, new generation UHR OCT system, achieving considerably improved visualization of intraretinal layers, especially the photoreceptor layer. main outcome measures: UHR OCT tomograms visualizing intraretinal differences in morphology of AOFVD and SD/FF as location and extension of deposits and loss of photoreceptors. Central foveal thickness defined as distance between internal limiting membrane and photoreceptors/retinal pigment epithelium interface. RESULTS: Patients with AOFVD had a mostly intact photoreceptor layer, a central foveal thickness of 142 +/- 23 microm as well as subretinal deposits. Patients with SD generally had a diffuse degenerative change with a visible reduction in thickness of all intraretinal layers, resulting in a corresponding reduction of central foveal thickness (94 +/- 38 microm) and central loss of photoreceptors (PRs). Comparative central foveal thickness of patients with AOFVD and SD/FF was significantly different (P < .001). Patients with FF had pigment epithelial deposits and paracentral focal photoreceptor loss. CONCLUSIONS: UHR OCT is a clinically feasible tool for examining intraretinal changes, in particular photoreceptor atrophy in macular dystrophies and, therefore, has the potential to be an adequate imaging system for monitoring the course of disease. PMID- 16376640 TI - Conductive keratoplasty: histological study of human corneas. AB - PURPOSE: To determine the morphologic changes in human corneas over time following radiofrequency-based conductive keratoplasty (CK) treatment. DESIGN: Prospective, observational case series. METHODS: In a single-center study six human corneas of six patients with localized peripheral keratoconus underwent CK treatment followed by penetrating keratoplasty. Three spots were applied in the periphery of each cornea (6 mm optical zone). Corneal buttons were examined with light and electron microscopy at different postoperative intervals up to 6 months post-CK. RESULTS: In samples assessed on day one post-CK, small areas of detachment between the basal layer of epithelial cells and Bowman's layer were observed. At 1 week after the CK procedure, the epithelium appeared almost normal. Endothelium and Descemet's membrane had no alterations. In all samples, thermally induced misconfiguration of collagen fibers, described as "crumpled" changes of collagen layers, was observed reaching 75% to 80% of the stromal depth. The area of alterations had a cylindrical shape with a diameter of 120 mum. CONCLUSIONS: The conductive keratoplasty procedure produced collagen "crumpling and splitting" changes in human corneas, which were observed during the follow-up of 6 months. Areas adjacent to treatment site were minimally damaged. PMID- 16376641 TI - Characteristics of keratoconus and pellucid marginal degeneration in mean curvature maps. AB - PURPOSE: To characterize keratoconus and pellucid marginal degeneration (PMD) on mean curvature maps. DESIGN: Observational case series. METHODS: A retrospective analysis of clinical records of 19 keratoconus eyes and eight PMD eyes. Keratometric mean curvature maps were computed from a placido-disk-based corneal topography system. The peak location and amplitude of distortions were quantified by fitting two-dimensional Gaussian functions. Computer simulations of keratoconic corneal topography were used to help interpret the results. RESULTS: Keratoconus and PMD eyes had a characteristic localized increase in convexity (cone) on mean curvature maps. Computer simulations showed that the peak on the mean curvature map accurately represented the peak of the cone-like distortion. However, the peak location and appearance of the same conic distortion on axial and tangential maps were greatly influenced by coexisting astigmatism. The Gaussian function provided an excellent model of keratoconus on mean curvature maps, with a mean cross-correlation of 0.80. According to Gaussian fitting, the vertical cone peak locations were -1.10 +/- 0.43 mm (mean +/- SD) in keratoconus eyes and -1.94 +/- 0.53 mm in PMD eyes (P < .01). The locations of peaks on axial and tangential maps differed considerably from those on mean curvature maps. Advanced keratoconus tended to have higher cones (amplitude 13.31 +/- 6.87 diopters) than early keratoconus (amplitude 10.65 +/- 1.56 diopters). CONCLUSIONS: Gaussian fitting of mean curvature maps accurately quantifies the peak location and amplitude of the cone in keratoconus and PMD eyes. This new topographic analysis might be useful in the diagnosis and tracking of corneal ectatic diseases. PMID- 16376642 TI - Choroidal biopsies for intraocular tumors of indeterminate origin. AB - PURPOSE: To evaluate the role of pars plana vitrectomy-assisted incisional biopsies in the management of choroidal tumors of unclear origin. DESIGN: Retrospective, noncomparative, consecutive interventional case series. METHODS: Ten consecutive patients with indeterminate choroidal tumors underwent a standardized three-port pars-plana vitrectomy-assisted subretinal biopsy using a bimanual approach with standard intraocular forceps and a diamond knife. Specimens were fixed in formaldehyde embedded in paraffin and further subjected to histopathological and immunohistochemical analyses. RESULTS: A histologic diagnosis was obtained in all (10 of 10) cases including choroidal melanoma (five of 10), metastasis (two of 10), subretinal hemorrhage (two of 10), and nodular scleritis (one of 10). Five eyes were enucleated as a result of the histologic diagnosis. Three cases of postoperative complications were seen in three patients (newly formed rhegmatogenous retinal detachment, increased serous retinal detachment, and vitreous hemorrhage). No cases of intra- or extraocular tumor spread were detected through follow-up periods ranging from 3 to 29 months. CONCLUSIONS: Pars plana vitrectomy-assisted incisional biopsy is a valuable diagnostic procedure for cases of choroidal tumors of unknown origin in selected patients. However, the relatively high frequency of postoperative complications noted in the present study and the potential risk of dissemination of tumor cells underscores the importance of rigorous case selection. PMID- 16376643 TI - Pediatric Amblyopia Risk Investigation Study (PARIS). AB - PURPOSE: To assess the learning curve, testability, and reliability of vision screening modalities administered by pediatric health extenders. DESIGN: Prospective masked clinical trial. METHODS: Two hundred subjects aged 3 to 6 underwent timed screening for amblyopia by physician extenders, including LEA visual acuity (LEA), stereopsis (RDE), and noncycloplegic autorefraction (NCAR). Patients returned for a comprehensive diagnostic eye examination performed by an ophthalmologist or optometrist. RESULTS: Average screening time was 5.4 +/- 1.6 minutes (LEA), 1.9 +/- 0.9 minutes (RDE), and 1.7 +/- 1.0 minutes (NCAR). Test time for NCAR and RDE fell by 40% during the study period. Overall testability was 92% (LEA), 96% (RDE), and 94% (NCAR). Testability among 3-year-olds was 73% (LEA), 96% (RDE), and 89% (NCAR). Reliability of LEA was moderate (r = .59). Reliability of NCAR was high for astigmatism (Cyl) (r = .89), moderate for spherical equivalent (SE) (r = .66), and low for anisometropia (ANISO) (r = .38). Correlation of cycloplegic autorefraction (CAR) with gold standard cycloplegic retinoscopic refraction (CRR) was very high for SE (.85), CYL (.77), and moderate for ANISO (.48). CONCLUSIONS: With NCAR, physician extenders can quickly and reliably detect astigmatism and spherical refractive error in one-third the time it takes to obtain visual acuity. LEA has a lower initial cost, but is time consuming, moderately reliable, and more difficult for 3-year-olds. Shorter examination time and higher reliability may make NCAR a more efficient screening tool for refractive amblyopia in younger children. Future study is needed to determine the sensitivity and specificity of NCAR and other screening methods in detecting amblyopia and amblyopia risk factors. PMID- 16376644 TI - Imaging polarimetry in central serous chorioretinopathy. AB - PURPOSE: To evaluate a noninvasive technique to detect the leakage point of central serous chorioretinopathy (CSR), using a polarimetry method. DESIGN: Prospective cohort study. METHODS: SETTING: Institutional practice. PATIENTS: We examined 30 eyes of 30 patients with CSR. MAIN OUTCOME MEASURES: Polarimetry images were recorded using the GDx-N (Laser Diagnostic Technologies). We computed four images that differed in their polarization content: a depolarized light image, an average reflectance image, a parallel polarized light image, and a birefringence image. Each polarimetry image was compared with abnormalities seen on fluorescein angiography. RESULTS: In all eyes, leakage area could be clearly visualized as a bright area in the depolarized light images. Michelson contrasts for the leakage areas were 0.58 +/- 0.28 in the depolarized light images, 0.17 +/ 0.11 in the average reflectance images, 0.09 +/- 0.09 in the parallel polarized light images, and 0.11 +/- 0.21 in the birefringence images from the same raw data. Michelson contrasts in depolarized light images were significantly higher than for the other three images (P < .0001, for all tests, paired t test). The fluid accumulated in the retina was well-visualized in the average and parallel polarized light images. CONCLUSIONS: Polarization-sensitive imaging could readily localize the leakage point and area of fluid in CSR. This may assist with the rapid, noninvasive assessment of CSR. PMID- 16376645 TI - Antioxidant supplements prevent oxidation of cysteine/cystine redox in patients with age-related macular degeneration. AB - PURPOSE: Determine whether antioxidant supplements alter the plasma glutathione and/or cysteine redox potential in age-related macular degeneration (AMD) patients. DESIGN: This was an ancillary study to the Age-Related Eye Disease Study (AREDS), where subset of AREDS subjects at two sites were studied at two time points, an average of 1.7 and 6.7 years after enrollment. METHODS: Plasma glutathione (GSH), glutathione disulfide (GSSG), cysteine (Cys), and cystine (CySS) were measured by high-performance liquid chromatography, and redox potentials of GSH/GSSG (E(h) GSH) and Cys/CySS (E(h) Cys) were calculated. The means of the metabolites and redox potentials were compared by repeated-measures analysis of variance for subjects receiving antioxidants and those not receiving antioxidants. RESULTS: At the first blood draw, the means for the antioxidant group (n = 153) and no antioxidant group (n = 159) were not significantly different for any of the metabolites or redox potentials. At the second draw, the GSH parameters were not significantly different between the antioxidant (n = 37) and no antioxidant (n = 45) groups; however, mean Cys was significantly higher in the antioxidant group (9.5 vs 7.2 micromol/l, P = .008). Also, mean E(h) Cys was significantly more reduced in the antioxidant group (-74 vs -67.3 mV, P = .03). CONCLUSIONS: The AREDS antioxidant supplements reduced oxidation of E(h) Cys but had no effect on GSH. Because Cys is important for cell growth, apoptosis, and immune function, the beneficial effect of antioxidant supplementation on progression to advanced AMD may be partially explained by its effect on E(h) Cys and/or its effect on Cys availability. PMID- 16376646 TI - Reliability of transport medium in the laboratory evaluation of corneal ulcers. AB - PURPOSE: To compare the microbiological yield of corneal ulcer cultures established by direct inoculation of culture media vs indirect inoculation by means of transport medium (Amies without charcoal). DESIGN: Single masked, prospective clinical trial. METHODS: Scrapings were obtained for Gram and potassium hydroxide (KOH) stains from eyes with presumed infectious keratitis and cultured by direct plating onto standard media. Samples were also held in transport media (Amies without charcoal) at room temperature and then plated after 4 and 24 hours. Yields from direct plating vs cultures by means of transport media were compared. RESULTS: Of 100 consecutive eyes examined with presumed infectious keratitis, Gram or KOH stain revealed a bacterial or fungal agent in 69 cases (69%). Of these, 26 were bacterial and 43 fungal. Twenty-two bacterial infections produced positive cultures by direct plating, and all produced the same organism with Amies medium after 4 and 24 hours, respectively. For 43 fungal infections identified by KOH stain, 29 (67%) yielded a positive result after 4 hours in Amies transport medium and 27 (63%) after 24 hours in Amies medium. A total of three cases (7%) that showed fungal infection on KOH stain but did not yield organisms by direct plating did so after inoculation with Amies transport medium. For all comparisons, there was no difference in recovery rates by means of transport medium compared with direct plating (McNemar exact P > .05). CONCLUSIONS: In the clinical setting, Amies transport medium may be a useful alternative to direct inoculation onto blood agar for the laboratory evaluation of infectious keratitis. PMID- 16376647 TI - Keratoprosthesis: the Dohlman-Doane device. AB - PURPOSE: To determine the usefulness of the Dohlman-Doane type I keratoprosthesis for visual rehabilitation in cases of poor prognosis for traditional penetrating keratoplasty. DESIGN: A retrospective, noncomparative interventional series of 25 patients who had sustained multiple graft failure or who were otherwise deemed poor candidates for conventional keratoplasty. METHODS: Candidates were evaluated for potential acuity, intraocular pressure, inflammation, the quality of the ocular surface, and overall prognosis for penetrating keratoplasty. The keratoprosthesis was assembled and mounted on an 8.5- or 9.0-mm diameter donor corneal button and sutured into an 8.0- or 8.5-mm diameter recipient bed with 12 to 16 9-0 nylon sutures. Patients were examined on the first day and subsequently in 1 week and 1 month and then at 3-month intervals. RESULTS: All devices were retained without dislocation or extrusion. There were no instances of endophthalmitis or surface infection. Fundus details were visible on the first postoperative day. Patients achieved their best acuity in an average of 2 months (range, 1 to 180 days). Improvement in acuity was observed in an average of 13 days (range, 1 to 60 days). Retroprosthetic membranes occurred in three cases, with multiple recurrences in one instance. Visual acuity ranged from no light perception to 20/25. CONCLUSION: We conclude that this prosthesis can be implanted routinely and maintained with minimal complications in poor prognosis keratoplasty, which presents the potential for visual rehabilitation. PMID- 16376648 TI - Laboratory model of tissue adhesive (2-octyl cyanoacrylate) in sealing clear corneal cataract wounds. AB - PURPOSE: To determine the efficacy of a tissue adhesive (2-octyl cyanoacrylate with parabens; Liquid Bandage, Johnson & Johnson, Skillman, New Jersey) in sealing clear corneal cataract wounds. DESIGN: Laboratory investigation. METHODS: Seven human donor globes were prepared for Miyake video microscopy. A 3.0-mm clear corneal incision was created. A transscleral cannula was inserted and connected to a bottle of saline. The bottle height was varied to alter intraocular pressure. Droplets of India ink were placed on the wound. Main outcome measure was any influx of India ink into the anterior chamber as viewed through the Miyake system with intraocular pressure (IOP) fluctuation or with manual pressure. If India ink was present in the eye, it was irrigated out, and the experiment was repeated with IOP fluctuation and manual pressure after the application of 2-octyl cyanoacrylate to the wound. RESULTS: One eye demonstrated the presence of India ink inside the eye on IOP reduction to <5 mm Hg. Three eyes demonstrated the presence of India ink inside the eye with manual pressure. Three eyes did not leak with manual pressure or IOP variation. All seven eyes without glue leaked with exaggerated manual pressure at the wound edge. Of the seven eyes with tissue adhesive, none demonstrated influx of India ink with IOP variation or manual wound manipulation. CONCLUSIONS: Our laboratory model demonstrates that 2 octyl cyanoacrylate prevents the influx of ocular surface fluid independent of IOP and manual wound manipulation. Further investigations in clinical models are necessary to determine the future use of this adhesive barrier substance. PMID- 16376649 TI - Neuropilin-1 expression by endothelial cells and retinal pigment epithelial cells in choroidal neovascular membranes. AB - PURPOSE: To determine if vascular endothelial growth factor (VEGF) coreceptor neuropilin-1 (NP-1) is expressed in choroidal neovascularization (CNV) and to localize the expression. DESIGN: Laboratory investigation. METHODS: Six CNV membranes (CNVMs) obtained from patients with subfoveal CNV attributable to age related macular degeneration (AMD) underwent immunohistochemistry for VEGF receptor-2 (VEGFR-2) and NP-1. The positive cell types were identified by double staining with anticytokeratin, anti-CD31, and antismooth muscle actin (SMA). RESULTS: Immunohistochemical staining revealed positivity for VEGFR-2 and NP-1 in all six CNVMs. Both receptors were strongly expressed by new choroidal endothelial cell forming vessels and CD-31-positive cells in the nonvascular area. They were also expressed in the pigmented retinal pigment epithelial layer and by cytokeratin-positive, nonpigmented retinal pigment epithelium cells in the nonvascular area. The nonpigmented retinal pigment epithelium cells positive for VEGFR-2 and NP-1 were highly colocalized with SMA. CONCLUSIONS: The presence of both VEGF and NP-1 suggest that NP-1 may play a role in the evolution of CNV in AMD. PMID- 16376650 TI - Tilt and decentration of three-piece foldable high-refractive silicone and hydrophobic acrylic intraocular lenses with 6-mm optics in an intraindividual comparison. AB - PURPOSE: Intraindividual comparison of tilt and decentration of three-piece foldable intraocular lenses (IOLs) with 6-mm optics and different edge design and material. DESIGN: Prospective randomized study. METHODS: Twenty-five patients with senile cataract (group I) received a foldable silicone, sharp optic edge IOL in one eye and a silicone, rounded optic edge IOL in the other eye. Group II (n = 28) received the foldable silicone, sharp optic edge IOL in one eye and an acrylate, sharp optic edge IOL in the other eye. Scheimpflug photography was performed after the procedure with an anterior eye segment analysis system. Tilt and decentration of the IOL optic were measured 1 week, 6 months, and 12 months after the procedure. RESULTS: In group I, the foldable silicone, sharp optic edge IOL showed a mean optic tilt of 3.03 +/- 1.79 degrees and an optic decentration of 0.24 +/- 0.13 mm; the silicone, rounded optic edge IOL showed a tilt of 3.26 +/- 1.69 degrees and a decentration of 0.23 +/- 0.13 mm. In group II, the foldable silicone, sharp optic edge IOL showed an average tilt of 2.34 +/- 1.81 degrees and a decentration of 0.29 +/- 0.21 mm after 12 months; the acrylate, sharp optic edge IOL had a tilt of 2.32 +/- 1.41 degrees and a decentration of 0.24 +/- 0.10 mm. There were no significant differences in either group. CONCLUSION: The examined IOLs showed a stable position regarding tilt and decentration in the first 12 postoperative months, independent of material and edge design. Slight deviations from the optical axis might affect the outcome in aspheric or multifocal IOLs. PMID- 16376651 TI - Recovery of corneal subbasal nerve density after PRK and LASIK. AB - PURPOSE: To measure and compare the return of corneal innervation up to 5 years after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK). DESIGN: Prospective, nonrandomized clinical trial. METHODS: Eighteen eyes of 12 patients received PRK to correct a mean refractive error of -3.73 +/- 1.30 diopters, and 16 eyes of 11 patients received LASIK to correct a mean refractive error of -6.56 +/- 2.44 diopters. Corneas were examined by confocal microscopy before and at 1, 2, 3, and 5 years after the procedures. Subbasal nerve fiber bundles were measured to determine density (visible length of nerve/frame area) and expressed as micrometers per square millimeters. Differences were compared by Friedman's test and adjusted for multiple comparisons by the Student-Newman-Keuls procedure. RESULTS: After PRK, mean subbasal nerve density was reduced by 59% at 1 year (2764 +/- 1321 microm/mm(2) [+/-SD]) when compared with preoperative (6786 +/- 1948 microm/mm(2); P < .001). By 2 years, subbasal nerve density (6242 +/- 1763 microm/mm(2)) was not significantly different from density before PRK and remained unchanged to 5 years (5903 +/- 3086 microm/mm(2)). After LASIK, subbasal nerve density was reduced by 51%, 35%, and 34% at 1, 2, and 3 years, respectively (P < .001). By 5 years, subbasal nerves had returned to densities (4441 +/- 2819 microm/mm(2)) that were not significantly different from densities before LASIK (5589 +/- 2436 microm/mm(2)). CONCLUSION: Corneal subbasal nerve density does not recover to near preoperative densities until 5 years after LASIK, as compared with 2 years after PRK. PMID- 16376652 TI - Nasolacrimal duct obstruction classified by dacryoendoscopy and treated with inferior meatal dacryorhinotomy. Part I: Positional diagnosis of primary nasolacrimal duct obstruction with dacryoendoscope. AB - PURPOSE: To investigate the site of primary nasolacrimal duct obstruction (PNLDO) using a dacryoendoscope and nasal endoscope. DESIGN: Retrospective, observational case series. METHODS: After local anesthesia, dacryoendoscopy and nasal endoscopy were performed in 149 cases (138 Japanese subjects) of PNLDO. Membranous nasolacrimal duct obstruction was termed as lower PNLDO, and occlusion at sac/duct junction termed as higher PNLDO. RESULTS: The incidence of lower PNLDO was 26.8% (40 of 149 cases), and 109 cases (73.2%) were diagnosed with higher PNLDO. CONCLUSIONS: The dacryoendoscope is extremely useful to observe directly and evaluate the site of obstruction with accuracy in PNLDO. The higher and lower PNLDO may have a different etiologies. PMID- 16376653 TI - Nasolacrimal duct obstruction classified by dacryoendoscopy and treated with inferior meatal dacryorhinotomy: Part II. Inferior meatal dacryorhinotomy. AB - PURPOSE: To report the performance of a newly devised technique, inferior meatal dacryorhinotomy (iDR), for membranous PNLDO (lower PNLDO). DESIGN: Retrospective, nonrandomized clinical trial. METHODS: Forty cases (37 patients) who ranged in age from 42 to 89 years old with lower PNLDO were treated by iDR, which is intubation that is assisted by dacryoendoscope, nasal endoscope, and radio frequency scalpel. The obstructed site was incised with a radio-frequency scalpel without osseous ablation, after which silicone tubes were placed temporarily. RESULTS: Postoperative follow-up evaluation showed that the success rate was 87.5% after 6 months to 2 years. CONCLUSION: The iDR technique is minimally invasive and effective for lower PNLDO. PMID- 16376654 TI - A progressive anterior fibrosis syndrome in patients with postsurgical congenital aniridia. AB - PURPOSE: To report the characteristics of a newly recognized clinical entity in congenital aniridia that we have termed aniridic fibrosis syndrome. DESIGN: Interventional case series. METHODS: Retrospective chart review of 155 eyes in 80 patients with congenital aniridia was carried out to identify and characterize eyes that had anterior chamber fibrosis. Histopathologic evaluation was performed in three eyes. RESULTS: Seven eyes in six patients were identified to have aniridic fibrosis syndrome. All eyes had undergone previous intraocular anterior segment surgery, some eyes with multiple procedures. Seven eyes had undergone cataract surgery with posterior chamber intraocular lens; six eyes had undergone previous implantation of tube shunt devices, and four eyes had undergone previous penetrating keratoplasty. Clinically, the syndrome was characterized by a progressive retrolenticular and retrocorneal membrane that caused forward displacement of intraocular lenses. Surgical findings indicated that the fibrotic membrane also can involve the ciliary body and anterior retina. Histopathologic evidence from three eyes indicated that the extensive fibrotic tissue originated from the root of the rudimentary iris and entrapped the intraocular lens haptics. Endothelial decompensation that was subsequent to the formation of the aniridic fibrosis syndrome was seen in all eyes. CONCLUSION: Aniridic fibrosis syndrome is characterized by the development of a progressive anterior chamber fibrosis. A possible mechanism that promotes the formation of this fibrotic material may be the proximity or touching of intraocular devices on immature vessels in the rudimentary iris found in aniridia. Patients with aniridia with a history of penetrating keratoplasty, intraocular lenses, and tube shunts should be monitored for aniridic fibrosis syndrome; early surgical intervention is recommended. PMID- 16376655 TI - Results of penetrating keratoplasty for visual rehabilitation after Acanthamoeba keratitis. AB - PURPOSE: To assess the results of penetrating keratoplasty in quiet eyes after resolution of Acanthamoeba keratitis. DESIGN: A retrospective interventional case series. METHODS: Penetrating keratoplasty was undertaken in 13 eyes of 13 patients with an average age of 29 +/- 13 years and a history of Acanthamoeba keratitis that was diagnosed by culture and/or confocal microscopy between January 1995 and September 2004. All eyes were observed for at least 3 months (average, 5 months) after the discontinuation of antiamebic therapy that had been administered for at least 4.5 months. In vivo confocal microscopy was used to ensure that no residual amoeba were present before the operation. Two keratoplasties were combined with a valve insertion; five keratoplasties were combined with cataract extraction, and one keratoplasty was preceded by a ciliary laser ablation. RESULTS: Postoperative best-corrected visual acuity ranged from 20/40 to 20/15, with the exception of one eye with advanced glaucoma that did not improve from a preoperative vision of light perception. No eye experienced rejection or amoebic recurrences, and 12 grafts remained clear throughout the follow-up period (8 months to 9 years; median, 23 months). One graft failed at 4 months because of uncontrolled glaucoma. The eye was regrafted, and the graft remained clear during the 28 months of follow-up evaluation. Two eyes with preoperatively diagnosed glaucoma needed subsequent surgery to control their intraocular pressure. CONCLUSION: Penetrating keratoplasty for visual restoration after Acanthamoeba keratitis appears to have an excellent long-term prognosis, provided amoebic infection has resolved and concurrent glaucoma is controlled. PMID- 16376657 TI - Diagnostic biopsy of selected intraocular tumors. PMID- 16376658 TI - Vitrectomy in the treatment of uveitis. AB - PURPOSE: To assess the evidence that pars plana vitrectomy (PPV) is useful in improving vision, reducing disease activity, or ameliorating cystoid macular edema (CME) in patients with uveitis. DESIGN: Review of the literature. METHODS: A Medline search was conducted for relevant articles published in English, German, or French. Articles were analyzed for content and evidence level. RESULTS: A total of 44 interventional case series published between 1981 and 2005 were identified that included 1575 patients (1762 eyes). Evidence level was grade CII-3 indicating possibly improved clinical outcomes with fair or poor evidence. The average age of patients was 36 years with a median duration of uveitis before surgery of 48 months and a median follow-up of 1.9 years. Intermediate uveitis was present in 841 eyes. Cystoid macular edema and cataract were common co morbidities, and there were large numbers of additional surgical procedures. Visual outcomes in 39 articles were stated as improved in 708 eyes (68%), unchanged in 202 eyes (20%), and worsened in 124 eyes (12%). Reduction in systemic medication following PPV was reported in 25 studies. The median reported percentage of patients per study with CME was 36% preoperatively and 18% postoperatively. CONCLUSIONS: Based on the evidence in the literature, PPV is possibly relevant to the outcomes of improving vision and reducing inflammation and CME. Randomized, controlled, collaborative trials or hypothesis-based case series with precise outcome measures that incorporate control groups would improve the quality of evidence supporting PPV as an adjunct to the medical treatment of uveitis. PMID- 16376656 TI - Long-term follow-up of fixation patterns in eyes with central scotomas from geographic atrophy that is associated with age-related macular degeneration. AB - PURPOSE: To study whether fixation patterns changed over time in patients with central scotomas from geographic atrophy from age-related macular degeneration. DESIGN: Prospective cohort study. METHODS: setting: Institutional. patient or study population: Prospective natural history study of geographic atrophy included 34 eyes of 25 patients with baseline acuity between 20/80 and 20/200 and with subsequent follow-up. observation procedures: Baseline and annual follow-up visits included best-corrected visual acuity, scanning laser ophthalmoscope macular perimetry, reading rate, and clinical evaluation. main outcome measures: Location of eccentric preferred retinal locus for fixation (PRL). RESULTS: At baseline, 77% of study eyes had a PRL. At the final visit (median follow-up, 5.3 years), 91% of study eyes had a PRL, with 81% of the eyes retaining the baseline PRL location. Fixation with the scotoma to the right and fixation with the scotoma superior were the first and second most common fixation patterns, respectively. Reading rates of <50 words/min were present in 54% of eyes. Eyes fixating with the scotoma to the left tended to have lower reading rates than eyes fixating with right or superior patterns. CONCLUSION: Fixation with right pattern remained the most common fixation pattern, both in patients with a PRL at baseline and in patients who had a PRL during follow-up. Eyes with a PRL at baseline generally retained this pattern. The reading rate data suggest an advantage of fixation with right or superior pattern, rather than left. Reading rate declined further during follow-up in most patients. PMID- 16376659 TI - Medical professional liability insurance and its relation to medical error and healthcare risk management for the practicing physician. AB - PURPOSE: To review the history and current issues surrounding medical professional liability insurance and its relationship to medical error and healthcare risk management. DESIGN: Focused literature review and authors' experience. METHODS: Medical professional liability insurance issues are reviewed in association with the occurrence of medical error and the role of healthcare risk management. RESULTS: The rising frequency and severity of claims and lawsuits incurred by physicians, as well as escalating defense costs, have dramatically increased over the past several years and have resulted in accelerated efforts to reduce medical errors and control practice risk for physicians. Medical error reduction and improved patient outcomes are closely linked to the goals of the medical risk manager by reducing exposure to adverse medical events. Management of professional liability risk by the physician-led malpractice insurance company not only protects the economic viability of physicians, but also addresses patient safety concerns. CONCLUSIONS: Physician owned malpractice liability insurance companies will continue to be the dominant providers of insurance for practicing physicians and will serve as the primary source for loss prevention and risk management services. To succeed in the marketplace, the emergence and importance of the risk manager and incorporation of risk management principles throughout the professional liability company has become crucial to the financial stability and success of the insurance company. The risk manager provides the necessary advice and support requested by physicians to minimize medical liability risk in their daily practice. PMID- 16376660 TI - The impact of corneal allograft rejection on the long-term outcome of corneal transplantation. AB - PURPOSE: To examine the influence of corneal allograft rejection on the survival of penetrating corneal transplantation, to review the status of conventional therapies to improve graft survival, and to consider prospects for alternative approaches to reduce the impact of rejection. DESIGN: Perspective, including prospective, observational cohort study. METHODS: An examination of the literature on human corneal graft rejection and data from the Australian Corneal Graft Registry, reviewed in the context of clinical experience. RESULTS: Corneal graft outcome is not improving with era. The sequelae of inflammation, whether occurring before corneal transplantation or subsequently, exert a profound influence by predisposing the graft to rejection. Of the developments that have been instrumental in reducing rejection in vascularized organ transplantation, living-related donation is not an option for corneal transplantation. However, HLA matching may be beneficial and requires reassessment. The evidence base to support the use of systemic immunosuppressive agents in corneal transplantation is thin, and topical glucocorticosteroids remain the drugs of choice to prevent or reverse rejection episodes. Experimental approaches to local allospecific immunosuppression, including the use of antibody-based reagents and gene therapy, are being developed but may be difficult to translate from the laboratory bench to the clinic. CONCLUSIONS: Corneal allograft rejection remains a major cause of graft failure. High-level evidence to vindicate the use of a particular approach or treatment to prevent or treat corneal graft rejection is lacking. In the absence of extensive data from randomized, controlled clinical trials, corneal graft registers and extrapolation from experimental models provide some clinically useful information. PMID- 16376661 TI - Immunohistochemical analysis of the internal limiting membrane peeled with infracyanine green. AB - PURPOSE: To investigate whether the peeled internal limiting membrane (ILM) contains cellular retinal cell fragments, and to learn more about their possible origin. DESIGN: Experimental study. METHODS: ILM peeled from ten eyes during vitrectomy by infracyanine green (ICG) was studied immunohistochemically using the markers: GFAP, S-100, and vimentin. Five ILM specimens were from eyes with diabetic macular edema (DME), two from eyes with a macular hole, and three from eyes with persisting macular edema after retinal detachment surgery. RESULTS: In eight of the ten ILM specimens, we found GFAP-positive staining, indicating the presence of remnants of footplates from Muller cells or glial cells. Two ILM specimens were positive for S-100, indicating the presence of neural cells or ganglion cells. CONCLUSIONS: ILM peeled from the retina during vitrectomy using ICG may contain remnants of Muller cell footplates, neural cells, and ganglion cells. PMID- 16376662 TI - Progression of peripheral anterior synechiae after laser iridotomy. AB - PURPOSE: To determine the incidence and to characterize the risk factors of peripheral anterior synechiae (PAS) progression after laser iridotomy (LI). DESIGN: Retrospective observational cohort study. METHODS: The extents of pre- and postlaser PAS and ocular and systemic data were obtained from the charts of 54 primary angle-closure glaucoma patients (59 eyes), all of whom had undergone LI. RESULTS: PAS progressed in 19 of the 59 eyes (32.2%), during a mean follow-up period of 34.4 months (range, 6-95 months). Multivariate analysis showed that the presence of plateau iris (P = .024), and a high intraocular pressure 1 hour before LI (P = .042) were risk factors of PAS progression. CONCLUSION: Approximately one of three of the eyes showed PAS progression during a 3-year follow-up period after LI. The probability of progression was found to be higher in the eyes that exhibited plateau iris and in eyes that proved to be relatively unresponsive to medication before LI. PMID- 16376664 TI - Accuracy of the Lang II stereotest in screening for binocular disorders in 6-year old children. AB - PURPOSE: To assess the accuracy of the Lang II stereotest in screening for strabismus, amblyopia, and anisometropia in 6-year-old children. DESIGN: Cross sectional population-based study. METHODS: The Sydney Myopia Study examined 1765 6-year-old children (78.9% of eligible) who were identified by random cluster sampling of 34 schools in Sydney, Australia. Sensitivity and specificity of the Lang II stereotest was determined by best stereoacuity. Cycloplegic autorefraction, assessment of visual acuity, and ocular motility were conducted. RESULTS: Test sensitivity ranged from 21.4% for anisometropia (> or =1.0 diopter) to 31.3% for amblyopia. The detection rate for new cases of amblyopia ranged from 20% to 40%; the detection rate for new cases of strabismus was 30%. Specificity was >98% in all three conditions. Children with false-negative results included newly diagnosed cases of strabismus (14 of 25 children) or amblyopia (5 of 12 children). CONCLUSION: The Lang II stereotest, when used alone, has very limited value as a screening test of binocular dysfunction. PMID- 16376663 TI - Long anterior lens zonules in late-onset retinal degeneration (L-ORD). AB - PURPOSE: We report new findings of peripupillary iris atrophy and long anteriorly inserted zonules in a family with late-onset retinal degeneration (L-ORD). DESIGN: The proband was noted to have anterior segment findings not previously described in L-ORD, an autosomal dominant condition resulting in severe visual impairment. A mutation in the C1QTNF5 (CTRP5) gene is causal. We identified family members with anterior segment findings. METHODS: Family members were examined with slit-lamp biomicroscopy and psychophysical tests including dark adaptation and visual fields. Genetic testing for the C1QTNF5 mutation was carried out. RESULTS: In this family with a proven mutation in this gene, peripupillary iris atrophy and abnormally long anterior zonular insertions were present before retinal changes and visual loss. CONCLUSIONS: Anterior segment findings have not previously been reported and along with impaired dark adaptation may serve as an early marker for this condition thus facilitating counseling and possible therapeutic intervention. PMID- 16376665 TI - Treatment of corticosteroid dependent optic neuropathy with intravenous immunoglobulin. AB - PURPOSE: To report a patient with corticosteroid dependent optic neuropathy treated with intravenous immunoglobulin (IVIg). DESIGN: Interventional case report. METHODS: Records review. RESULTS: A 25-year-old woman developed bilateral vision loss with pain on eye movement in association with disk edema, dilated retinal veins, and hemorrhage. The vision improved markedly with prednisone, but she required a minimum of 20 mg/d despite concomitant therapy with methotrexate, cyclosporine, and/or mycophenolate mofetil. IVIg, 0.5 g/kg/d for 3 days each month with subsequent reduction in frequency, allowed discontinuation of all immunosuppression. CONCLUSIONS: Corticosteroid dependent optic neuropathy was successfully treated with IVIg. PMID- 16376666 TI - High-speed optical coherence tomography of laser iridotomy. AB - PURPOSE: To describe high-speed (4000 axial scans/s) optical coherence tomography (OCT) findings in a patient with narrow angles. DESIGN: Interventional case report. METHODS: A 56-year-old woman was found to have occludable narrow angles on OCT screening. This was confirmed by gonioscopy. Bilateral iridotomy was performed. Imaging of the angles was performed with a high-speed OCT prototype before and after iridotomy. RESULTS: OCT showed narrow angle bilaterally. Cornea, sclera, scleral spur, trabecula, iris, and iris recess were visualized. After iridotomy, the OCT showed reduction of iris concavity and patent iridotomies. Quantitative measurements of trabecula-iris space area on the OCT images showed widening of the angles to nonoccludable values. CONCLUSIONS: A 1.3-microm wavelength OCT allows noncontact quantitative assessment of the angle and may be useful in the management of narrow-angle glaucoma. PMID- 16376667 TI - Immunotherapy with imiquimod 5% cream for eyelid nodular basal cell carcinoma. AB - PURPOSE: To evaluate the efficacy and safety of topical imiquimod 5% cream for the treatment of eyelid basal cell carcinoma. DESIGN: Two interventional case reports. METHODS: Imiquimod 5% cream was applied topically once daily, 3 days a week for 8 to 12 weeks, in two patients affected by eyelid nodular basal cell carcinoma. Patients were followed up clinically with slit-lamp examination for evidence of tumor disappearance or recurrence, and local and systemic side effects. RESULTS: Complete clinical response was obtained in both patients. No severe local side effects were observed. Patients did not show any local recurrence after 1 year. CONCLUSIONS: Topical imiquimod 5% cream seems to be a useful treatment for eyelid nodular basal cell carcinoma in selected cases, but further long-term studies are needed to assess the efficacy and safety of this approach. PMID- 16376668 TI - Central corneal thickness and development of glaucomatous optic disk hemorrhages. AB - PURPOSE: To evaluate whether central corneal thickness influences the development of optic disk hemorrhages in chronic open-angle glaucoma. DESIGN: Prospective observational clinical study. METHODS: The study included 390 eyes of 223 white subjects with chronic open-angle glaucoma observed during a mean follow-up time of 61.3 +/- 36.4 months. Central corneal thickness was measured by corneal pachymetry. RESULTS: The event of optic disk hemorrhages during follow-up was detected in 63 eyes (16.2%). Development of optic disk hemorrhages was, univariately (P = .73) as well as in a multiple Cox regression analysis, controlling for age, sex, normal tension glaucoma, intraocular pressure, neuroretinal rim area, and size of beta zone of peripapillary atrophy, statistically independent (P = .56) of central corneal thickness. CONCLUSIONS: Development of optic disk hemorrhages may not be markedly influenced by central corneal thickness. PMID- 16376669 TI - Cytomegalovirus retinitis after intravitreous triamcinolone in an immunocompetent patient. AB - PURPOSE: To report the case of a 75-year-old man with diabetes who developed cytomegalovirus (CMV) retinitis after intravitreous injection of triamcinolone acetonide (IVTA). DESIGN: Observational case report. METHODS: Review of medical records. RESULTS: A 75-year-old man with diabetic macular edema developed arcuate retinal whitening after IVTA. A presumptive diagnosis of viral retinitis was made, and a vitrectomy was performed. Polymerase chain reaction of the vitreous was positive for CMV DNA. An infectious disease consultant found no signs of systemic CMV infection, and laboratory examination revealed that the patient was HIV negative. The patient responded well to intravitreal ganciclovir and oral valganciclovir, but when therapy was discontinued, the retinitis recurred and CMV DNA was again detected in the vitreous. The retinitis once again responded to antiviral therapy. CONCLUSIONS: CMV retinitis can occur after local immunosuppression with IVTA. Clinicians should be aware of this rare complication of IVTA. PMID- 16376670 TI - Anterior uveitis and iris nodules that are associated with Langerhans cell histiocytosis. AB - PURPOSE: To describe a case of Langerhans cell histiocytosis (LCH) that involved the anterior uveal tract. DESIGN: Interventional case report. METHODS: A retrospective review was conducted on a patient with iris nodules and anterior uveitis in the setting of LCH. Visual acuity and clinical findings that were noted on slit lamp biomicroscopy were extracted. RESULTS: An 18-year-old male patient with unilateral anterior segment inflammation and iris nodules experienced visual improvement from 20/200 to 20/25 after treatment with a 5-day course of topical corticosteroids. Regression of the iris nodules and anterior segment inflammation was also noted. Bone marrow aspirate confirmed recurrent, active LCH. CONCLUSION: The clinician should include LCH in the differential diagnosis when faced with anterior segment inflammation in conjunction with iris nodules. Additionally, LCH can be treated successfully with topical corticosteroid therapy. PMID- 16376671 TI - Changes of the lid fissure after surgery on horizontal extraocular muscles. AB - PURPOSE: To evaluate the effect of isolated medial and lateral rectus muscle surgery on eye lid position. DESIGN: Prospective, interventional case series. METHODS: We recruited 36 patients with strabismus with surgery performed on one horizontal extraocular muscle only. The width of the lid fissure and the position of the upper and lower lids in relation to the pupil center were measured before and 3 months after surgery. Regression analysis was used to evaluate the surgical effect. RESULTS: Recessions induced a widening of and resections a narrowing of the lid fissure, mainly because of changes of the lower lid position. The effect was statistically significant (change in lid fissure width, = 0.13 x surgical dose in millimeters; r(2) = .35; P = .0001). CONCLUSION: Patients should be informed about changes in lid fissure width after operations on horizontal extraocular muscles. If adequately integrated into the surgical plan, this side effect may alleviate pre-existing interocular differences in lid configuration. PMID- 16376672 TI - Accuracy of referencing in the ophthalmic literature. AB - PURPOSE: To investigate the frequency of citation and quotation errors in the ophthalmic literature. DESIGN: Analysis of 200 references from 100 papers published in 10 ophthalmic journals. METHODS: A sample of 20 references was randomly selected from each of the 10 journals and each reference was checked for accuracy. Quotations were categorized as totally, partially, or not accurate. RESULTS: There were 35 citation errors in 32 references, only four of which were errors in PubMed. Thirty quotations of references were not accurate; 20 were partially accurate. CONCLUSIONS: Citation and quotation errors are relatively common within the ophthalmic literature. This may be improved through technical editing. PMID- 16376673 TI - Amniotic membrane as an adjunct to donor sclera in the repair of exposed glaucoma drainage devices. AB - PURPOSE: To describe a novel method of repairing exposed glaucoma drainage devices (GDD) using amniotic membrane transplantation (AMT) in place of conjunctiva to cover donor sclera when conjunctival closure is hampered by adjacent scarring. DESIGN: Retrospective noncomparative interventional case series. METHODS: Consecutive case series of nine GDD erosions over 5 years repaired by a double-layer technique of scleral allograft plus AMT. RESULTS: Successful repair was achieved in seven cases. One GDD plate re-eroded after pars plana vitrectomy; a second leaked from the tube entry site. CONCLUSIONS: AMT is a safe and useful conjunctival substitute to cover donor sclera in situations where conjunctival scarring might otherwise preclude successful repair. PMID- 16376674 TI - Histopathologic examination of conjunctival tophi in gouty arthritis. AB - PURPOSE: To report a case of conjunctival tophi in a 59-year-old woman with gouty arthritis. DESIGN: Observational case report. METHODS: A 59-year-old woman with a 25-year history of severe gouty arthritis presented with bilateral chalky white conjunctival deposits. Conjunctival biopsies were obtained, and light and electron microscopy studies were performed. RESULTS: Light microscopy confirmed multiple gouty tophi. Transmission electron microscopy revealed intracytoplasmic crystalline deposits within histiocytes. Extracellular aggregates of the crystalline deposits were enclosed by membranous material. CONCLUSIONS: Conjunctival crystals may aid in identifying undiagnosed patients. In addition to conjunctival tophi, patients may also have uveitis, corneal crystals, and band keratopathy. PMID- 16376675 TI - Pars plana vitrectomy with gas tamponade for lamellar macular holes. AB - PURPOSE: To report anatomic and visual improvement after pars plana vitrectomy with gas tamponade for a lamellar macular hole with poor central visual acuity. DESIGN: Two interventional case reports. METHODS: Two patients with a lamellar macular hole underwent vitrectomy, internal limiting membrane peeling, and long acting gas injection. Main outcome measures included best-corrected visual acuity, biomicroscopic appearance, and optical coherence tomography findings. RESULTS: Vitrectomy with gas tamponade resulted in biomicroscopic, functional, and tomographic improvement in both patients for follow-up periods of 12 months. CONCLUSIONS: Vitrectomy with gas tamponade may be an effective method for a lamellar macular hole with poor visual acuity. PMID- 16376676 TI - Climatology of dematiaceous fungal keratitis. AB - PURPOSE: To explore how seasonal climate patterns may influence the epidemiology of human keratomycosis. DESIGN: Retrospective ecological study. METHODS: The seasonality of 52 episodes of dematiaceous fungal keratitis occurring along the Gulf of Mexico from 1972 to 2004 was examined. The temperature and relative humidity on the date of onset of 36 eyes with Curvularia keratitis was then compared with regional atmospheric conditions. RESULTS: A relative increase in the diagnosis of dematiaceous fungal keratitis occurred during late summer and persisted throughout autumn. The onset of Curvularia keratitis clustered in warm, humid months. CONCLUSIONS: Ocular phaeohyphomycosis, including curvulariosis, varies with seasonal weather in the subtropics. PMID- 16376677 TI - Retinopathy in persons with impaired glucose metabolism: the Australian Diabetes Obesity and Lifestyle (AusDiab) study. AB - PURPOSE: To describe risk factors and vascular disease associations of retinopathy in people with impaired glucose metabolism. DESIGN: Population-based cross-sectional study. METHODS: Retinal photographs taken of participants with impaired glucose tolerance and impaired fasting glucose were graded for retinopathy. Risk factors and cardiovascular outcomes were ascertained. RESULTS: Retinopathy was present in 69 (6.7%) of 1027 participants with impaired glucose tolerance or impaired fasting glucose. After multivariate adjustment, retinopathy was associated with prevalent stroke (odds ratio 4.2, 95% confidence interval, 1.8 to 9.7). CONCLUSIONS: Retinopathy is common in persons with impaired glucose metabolism and may be associated with prevalent stroke. PMID- 16376678 TI - Ankle-brachial index and the prevalence of age-related maculopathy. AB - PURPOSE: To examine the association of ankle-brachial blood pressure index (ABI) with prevalence of age-related maculopathy (ARM). DESIGN: A cross-sectional cohort study. METHODS: ABI was measured in 2447 subjects aged 53 to 97 years. ARM was determined from 30-degree color stereoscopic fundus photographs. RESULTS: Low ABI (< or =0.9) was present in 5.4% of subjects. Early ARM was present in 22.1% of subjects with and 18.8% without low ABI. Late ARM was present in 5.3% of subjects with and 1.7% without low ABI. This result was not statistically significant. CONCLUSIONS: Low ABI does not appear to be a risk factor for ARM. PMID- 16376679 TI - Performing vitreous biopsy by perfluorocarbon-perfused vitrectomy. AB - PURPOSE: To evaluate the safety and efficacy of perfluorocarbon-perfused vitrectomy (PCPV) as a novel technique in obtaining a large undiluted vitreous biopsy. DESIGN: Cross-sectional interventional study. METHODS: Patients with undiagnosed posterior uveitis scheduled for vitreous biopsy underwent PCPV. A syringe containing 5 ml of perfluorocarbon liquid (PCL) was connected to the infusion line. Aspiration of the central and superior vitreous was initiated with simultaneous infusion of the PCL. RESULTS: Twenty eyes of 20 patients were included in this study. The mean +/- SD amount of PCL used in each eye was 4.50 +/- 0.69 ml. The volume of vitreous sample obtained in each eye was 2.25 +/- 0.413 ml. No complications occurred. CONCLUSIONS: PCPV is a safe and effective method for obtaining large undiluted vitreous biopsy. PMID- 16376680 TI - Fundus autofluorescence imaging in serous and drusenoid pigment epithelial detachments associated with age-related macular degeneration. AB - PURPOSE: To investigate the fundus autofluorescence (FAF) imaging findings in patients with serous and drusenoid pigment epithelial detachment (PED) associated with age-related macular degeneration (AMD). DESIGN: Observational case series. METHODS: Fourteen eyes of 13 consecutive patients with serous (n = 10) and drusenoid (n = 4) PED were prospectively included. RESULTS: In all cases of serous vascularized (n = 7) and avascular (n = 3) PED, the FAF signal was increased, corresponding to the area of detachment. In drusenoid PED, the FAF was either increased or decreased. The decreased signal was recorded in cases associated with pigmented clumping. CONCLUSIONS: These findings are useful for a more detailed phenotyping of patients with PED associated with AMD. FAF may derive not only from lipofuscin, but also from other sources, such as sub-retinal pigment epithelium fluid. PMID- 16376681 TI - External (subciliary) vs internal (transconjunctival) involutional entropion repair. PMID- 16376683 TI - Natural history of asymptomatic clinical retinal detachments. PMID- 16376684 TI - Scleral canal size in patients with optic nerve drusen. PMID- 16376686 TI - Natural history of asymptomatic clinical retinal detachments. PMID- 16376689 TI - Natural history of asymptomatic clinical retinal detachments. PMID- 16376691 TI - Postoperative corneal swelling correlates strongly to corneal endothelial cell loss after phacoemulsification cataract surgery. PMID- 16376693 TI - The cytotoxicities induced by PM 10 and particle-bound water-soluble species. AB - A 1-year field sampling of PM(10) was performed at a town that usually has the worst air quality in Taiwan to examine if PM(10) is a good indicator for pollutant-induced cytotoxicity. The average PM(10) concentration in summer was the lowest, while the other three seasons did not show statistical difference in their PM(10) means. The pollutant-induced cytotoxicity presented as the cumene hydroperoxide equivalent concentration (CEC) was found to positively correlate with PM(10) concentrations and this study yielded a yearly average of the seasonal CEC 12.+/-8.54 microM with the magnitudes in sequence for the four seasons as: fall>winter>spring>summer. Positive relationship was also found between seasonal PM(10) and their corresponding CECs. The exponential regression model obtained from this study shows: CEC=3.305 exp(0.0118 PM(10)) (R(2)=0.634). The CEC correlates more significantly with NO(3)(-), SO(4)(2-), NH(4)(+) and Cl( ) (secondary aerosol species) than with the Na(+), K(+), Ca(2+) and Mg(2+) (crust related species) in PM(10). However, the best multivariable model obtained from this study to relate CEC with the concentrations of PM(10)-bearing water-soluble species shows: CEC=exp(1.4751+0.0470[SO(4)(2-)]+0.0143[NO(3)(-)]) (R(2)=0.550). PMID- 16376694 TI - Effect of maternal manganese blood levels on erythrocyte calcium-pump activity in newborns. AB - Manganese (Mn) is widely distributed in the biosphere but occurs in only trace amounts in animal tissues. Although Mn deficiency and toxicity both have pathological consequences, the underlying biochemical lesions have not been well defined. In vitro studies suggest that transport proteins are affected by Mn, lead (Pb), and selenium (Se). Among these transport proteins, the calmodulin regulated calcium pump (Ca(2+)Mg(2+)ATPase) could be inhibited by Mn. In order to understand Mn biochemical pathways, we examined the relationships between Mn blood levels and red blood cell Ca-pump activity among 248 mothers and newborns, environmentally exposed to Mn, Pb, and Se. POPULATION AND METHODS: 248 mother newborn pairs were recruited at Robert Debre University Hospital (Paris). Blood Mn and Pb concentrations were measured by absorption spectrophotometry. Se was measured by fluorometric method. Red blood cell membrane suspensions were obtained for Ca-pump activity measurements. Linear and quadratic regression models and Pearson correlation were performed. RESULTS: A non-linear parabolic relationship between maternal Mn blood levels and newborn Ca-pump activity was discovered from the analysis of the observed data. The peak level of maternal Mn that corresponded to a maximal activity of the newborn Ca-pump was estimated at 23.9 microg/l with a 95% confidence interval of 17.6 to 32.4 microg/l. An inhibition of this enzyme was observed at low and high levels of maternal Mn. The relationships between the newborn Ca-pump activity and maternal Se and Pb levels became non-significant after adjustment on all the co-factors included in the final model. CONCLUSION: Maternal environmental exposure to Mn, as reflected by maternal blood levels of this metal, is associated with a reduced activity of newborn erythrocyte Ca-pump in a non-linear pattern. Mn levels between 17.6 and 32.4 microg/l in maternal blood probably correspond to the optimal physiological concentration for the metabolism of this enzyme in newborns. PMID- 16376695 TI - Considerations for improving the accuracy of exposure to disinfection by-products by ingestion in epidemiologic studies. AB - Disinfection by-product (DBP) exposure characterization studies are often based on the analysis of a limited number of samples collected from a distribution system (DS) in which DBP levels are variable over time and space. A compositing technique was developed to simplify the sample collection procedures for integrating over temporal variations in DBPs measured in terms of trihalomethanes (THMs), haloacetic acids (HAAs), and total organic halogen (TOX). Over the course of 5 days analysis, the single composited sample was within 94-100% of the average THM concentration in all grab samples, 92-105% of HAAs, and 130% of the TOX concentration. Additionally, temporal variability factors such as timing of sample collection and the handling of tap water prior to consumption were found to influence DBP levels in consumers' drinking water. Included in our study of home water use are the effects of boiling which removed up to 98% of THMs and point of use (POU) devices which all showed DBP removal but differed depending on the device used. These factors should be taken into consideration in DBP exposure characterization for epidemiologic studies. PMID- 16376696 TI - The use of town refuse ash in urban agriculture around Jos, Nigeria: health and environmental risks. AB - This paper reports on a study that examines the health and environmental risks of using town refuse ash in urban vegetable production in Jos, Nigeria, in terms of heavy metal accumulation in the food chain. Soil and crop samples, collected from five study farms, and samples of the river water used for irrigation, were analysed for seven heavy metals Fe, Mn, Zn, Cu, Ni, Cd and Pb. On the basis of the field data the paper discusses: (1) the potential soil deficiencies and toxicities; (2) the probable links between soil heavy metal levels and fertilisation practices; (3) the heavy metal concentrations in crop tissue in relation to crop growth and human health. The findings suggest that soil concentrations of the seven metals fall within 'typical' soil levels, and that there should not be any problems of either toxicities or deficiencies for crop growth. There was evidence of slight accumulation of Zn, Cu and Cd on some of the farms with a history of town refuse ash use. However, in all farms lettuce crops contained very large concentrations of Fe, and Pb concentrations that were 20 to 40 times higher than the WHO/FAO maximum recommended level in leafy vegetables for human consumption. The Cd content of carrot tissue was 10 times higher than the WHO/FAO recommended limit. The relatively small number of soil and crop samples precluded any formal attempt at correlating the concentrations of heavy metals found in the vegetable crops with the farm levels. Nevertheless, the data suggested that these were not linked. The paper goes on to consider various potential sources of the metals found in the crops, including irrigation water, town refuse ash and air-borne dust, and discusses additional health and environmental risks pertaining to the use of town refuse ash. Undoubtedly, the heavy Pb and Cd contamination of certain crops indicates the urgent need for future studies to ascertain the precise source of these metals, and although the practice of using town refuse ash does not appear to have resulted in large-scale contamination of soil in the farming area, there are a number of unsafe practices associated with it that call for the identification of strategies for the safe utilisation of urban waste in Jos. PMID- 16376697 TI - Association of polychlorinated biphenyls (PCBs) with live algae and total lipids in rivers-a field-based approach. AB - The association of PCBs and live algal cells in rivers was studied at four locations during four seasons in two Wisconsin rivers. Positive relations between particle-associated PCBs and both chlorophyll-a and algal carbon concentrations indicated that live algal cells were a significant sorption phase for dissolved PCBs. Large Pennate diatoms (Navicula, Synedra, Pinnularia, Diatoma, and Cocconeis), or more rarely, Euglenoids (Trachelomonas sp.), dominated most sample assemblages on an algal carbon basis. These assemblages made up the highest percentage of total SOC during spring (average=50%) and lowest during summer (average=15%). At the three impounded sites, most individual PCB congeners were relatively enriched in samples characterized by: (1) high concentrations of algal carbon (as a percent of SOC), (2) algal assemblages dominated (or co-dominated) by Euglenoids, and (3) high concentrations of total lipids. Despite relatively higher masses of sorbed PCBs in the most lipid-rich samples, there was no robust correlation between total lipid content and particle-associated PCBs when aggregating all samples from the study. A possible explanation is that PCBs are associated with other structural components in live algae and (or) departure from chemical equilibrium in the river due to algal growth kinetics. A kinetic uptake model was used to calculate the mass of PCBs associated with the total organic carbon content of live algae. Based on this model, PCBs were enriched in algal cells during bloom seasons (spring and fall) compared to non-bloom seasons (summer and winter). Further, although individual PCB congener partition coefficients (log) to live algal cells (range=5.3-6.4) overlapped to those for detritus (range=3.6-7.4), PCBs tended to be enriched in detrital carbon pools during non-bloom conditions. The larger range of estimated PCB partition coefficients for detritus likely reflects the more heterogeneous nature of this material compared to live algal cells. PMID- 16376698 TI - The distribution and transport of Sellafield derived 137 Cs and 241 Am to western Irish Sea sediments. AB - Sellafield derived (137)Cs and (241)Am were analysed in surficial sediments at 23 sites in the western Irish Sea. Concentrations varied between 3-175 Bq (137)Cs/kg and 1-147 Bq (241)Am/kg. There are two distinct basins in the western Irish Sea separated by an area of restricted depth and this bathymetry is reflected in the radionuclide concentrations. Highest concentrations were found in the northerly samples and substantially lower concentrations further south. This pattern of distribution can be explained by the predominantly northerly residual flow and the existence of seasonal stratification in this area of weak tides. (137)Cs/(241)Am ratios were also highest in the northern basin, and it is suggested that this is caused by differences in the mechanisms of transport of the two elements. The direct physical migration of contaminated particles from the eastern Irish Sea is the dominant transport mechanism in the north of the study area, whereas transport in the dissolved phase is most important to the southern region. Concentrations of both elements were best correlated with the percentage <15 microm particulate fraction suggesting that particles within this range are most important for direct transport and scavenging from the soluble phase. PMID- 16376699 TI - High background radiation areas of Ramsar in Iran: evaluation of DNA damage by alkaline single cell gel electrophoresis (SCGE). AB - The hot springs in special areas in Ramsar, a northern coastal town in Iran, contain (226)Ra and (222)Rn. The natural radiation effects, radiosensitivity or adaptive responses, on the inhabitants of high natural radiation in Ramsar were studied. The single cell gel electrophoresis was used to monitor DNA damages. Three groups of volunteers were selected, one from high natural background radiation areas as the case group and two from normal background radiation areas as controls (control 1 and control 2). The latter one had the similar living situation to case group while the other (control 2) had different living situation from the other groups. Peripheral blood mononuclear cells (PMNCs) were separated and irradiated by (60)Co source at five different gamma doses. It was found that the spontaneous level of DNA damage and the induced DNA damage in all challenging doses in case group was considerably higher than control groups (p < 0.05). On the other hand, the repair rate in those volunteers, who received less than 10.2 mSv/y was significantly more than the control groups. In the contrary, individuals who live in homes with more than 10.2 mSv/y had incomplete repair. Additionally the plasma and urinary levels of vitamin C were measured spectrophotometrically. Although the concentration of vitamin C of plasma was equal in case and control 1 groups, the urinary level of vitamin C was found to be lower in the case group. PMID- 16376700 TI - Lifestyle risk factors predict healthcare costs in an aging cohort. AB - BACKGROUND: While the U.S. elderly population uses a disproportionate amount of healthcare resources, there is limited knowledge from prospective studies regarding the impact of lifestyle-related factors on costs in this group. The association was examined between smoking, drinking, exercise, body mass index (BMI), and changes in these risk factors, and healthcare costs after 4 years among 68- to 95-year-olds. METHODS: A total of 1323 participants completed annual surveys providing information on lifestyle factors (1986-1994) and health utilization (1994-1998). Healthcare costs in nine categories were ascertained from validated utilization. The relationships between risk factors and costs were examined in 2004 using linear regression models. RESULTS: Fewer cigarette pack years and lower BMI were the most significant predictors of lower total costs in 1998 (p<0.001), controlling for baseline sociodemographic factors, costs, and conditions. Associations with smoking were strongest for hospitalizations, diagnostic tests, and physician and nursing-home visits. Those who reduced smoking by one pack per day experienced cost savings of 1160 dollars (p<0.05). The costs for normal weight compared to minimally obese seniors were approximately 1548 dollars lower, with diagnostic testing, physician visits, and medications accounting for much of this difference. Daily walking, measured at baseline, also predicted lower costs for hospitalizations and diagnostic testing. CONCLUSIONS: Seniors who were leaner, smoked fewer cigarettes over a lifetime, reduced their smoking, or walked farther had significant subsequent cost savings compared to those with less-healthy lifestyle-related habits. PMID- 16376701 TI - Racial/ethnic disparities in the use of preventive services among the elderly. AB - OBJECTIVES: Minorities have worse health outcomes compared to whites, which are partially explained by racial/ethnic disparities in use of health services. Less well known, however, are whether these disparities persist among the elderly, the only group that possesses near universal health insurance coverage by Medicare, and how variation in Medicare coverage affects the receipt of preventive services. The scope of racial/ethnic disparities in the use of preventive services in the elderly was assessed, and the impact of the type of health insurance coverage on the use of preventive services was measured. METHODS: Data were derived from the 2001 California Health Interview Survey, a random-digit dial population-based survey, collected between November 2000 and October 2001. Analysis for this project was conducted in 2004. The association of race/ethnicity and type of health insurance with receipt of preventive services was assessed using bivariate and multivariate logistic regression models. RESULTS: African Americans and Latinos were significantly less likely to be vaccinated for influenza, and Asian Americans were significantly less likely to obtain a mammogram compared to whites, while controlling for other explanatory factors. Moreover, those with Medicare plus Medicaid coverage were significantly less likely to use all four preventive services compared to those with Medicare plus private supplemental insurance. CONCLUSIONS: Despite near-universal coverage by Medicare, racial/ethnic disparities in the use of some preventive services among the elderly persist. Further research should focus on identifying potential cultural and structural barriers to receipt of preventive services aimed at designing effective intervention among high-risk groups. PMID- 16376702 TI - Trends in care practices and outcomes among Medicare beneficiaries with diabetes. AB - BACKGROUND: While diabetes is a major issue for the aging U.S. population, few studies have described the recent trends in both preventive care practices and complications among the Medicare population with diabetes. Using the Medicare Quality Monitoring System (MQMS), this 2004 study describes these trends from 1992 to 2001 and how these rates vary across demographic subgroups. METHODS: Outcomes include age- and gender-adjusted rates of 15 indicators associated with diabetes care from 1992 to 2001, the absolute change in rates from 1992 to 2001, and 2001 rates by demographic subgroups. The data were cross-sectional samples of Medicare beneficiaries with diabetes from 1992 to 2001 from the Medicare 5% Standard Analytic Files. RESULTS: Use of preventive care practices rose from 1992 to 2001: 45 percentage points for HbA1c tests, 51 for lipid tests, 8 for eye exams, and 38 for self-monitoring of glucose levels (all p<0.05). Rates for short term and some long-term complications of diabetes (e.g., lower-extremity amputations and cardiovascular conditions) fell from 1992 to 2001 (p<0.05). However, rates of other long-term complications such as nephropathy, blindness, and retinopathy rose during the period (p<0.05). Nonwhites and beneficiaries aged <65 and >85 exhibited consistently higher complication rates and lower use of preventive services. CONCLUSIONS: The Medicare program has seen some significant improvement in preventive care practices and significant declines in lower-limb amputations and cardiovascular conditions. However, rates for other long-term complications have increased, with evidence of subgroup disparities. The MQMS results provide an early warning for policymakers to focus on the diabetes care provided to some vulnerable subgroups. PMID- 16376703 TI - Comparison of racial/ethnic disparities in adult immunization and cancer screening. AB - BACKGROUND: Racial/ethnic disparities in adult influenza and pneumococcal vaccination are marked and poorly understood. The purpose of this study was to contrast these disparities with disparities in other clinical preventive services -mammography and colorectal cancer screening--that are targeted to older populations. METHODS: Data from the 2000 National Health Interview Survey were analyzed in 2004 to determine to what degree race/ethnicity remains a predictor of the receipt of each service after adjusting for personal and health characteristics, socioeconomic status (SES), and access to and utilization of care variables. RESULTS: Blacks and Hispanics were significantly less likely to report receipt of nearly all preventive services examined. Among whites, 57%, 67%, 67%, and 40% reported pneumococcal vaccination, influenza vaccination, mammography, and colorectal cancer screening, respectively. Among blacks, those proportions were 31%, 48%, 60% and 33%, respectively; among English-speaking Hispanics, 35%, 60%, 60%, and 30%, respectively; and among Spanish-speaking Hispanics, 24%, 49%, 52%, and 19%, respectively. After adjusting for personal and health characteristics, socioeconomic factors, and measures of access to and utilization of care, blacks and English- and Spanish-speaking Hispanics remained significantly less likely than whites to report the receipt of pneumococcal vaccination; blacks remained significantly less likely to report influenza vaccination than whites; and Spanish-speaking Hispanics remained significantly less to report colorectal cancer screening than whites. CONCLUSIONS: Most racial/ethnic disparities seen in breast and colorectal cancer screening are explained by differences in SES. In contrast, racial/ethnic disparities in adult immunization persist, and especially for pneumococcal vaccination, suggesting that different barriers may be involved. PMID- 16376704 TI - Influenza and pneumococcal vaccination of adults aged > or = 65: racial/ethnic differences. AB - BACKGROUND: Influenza and pneumococcal polysaccharide vaccination (PPV) rates among persons aged > or = 65 years are significantly below national objectives of 90%, particularly among blacks and Hispanics. This study of the 2002-2003 influenza season examines factors that may be associated with low coverage. METHODS: A national sample of 1839 community-dwelling adults aged > or = 65 years was surveyed by telephone during January-May 2003. Outcomes analyzed in 2004-2005 included self-reported influenza vaccination and PPV; place of vaccination; and among the unvaccinated, main reasons for nonvaccination, awareness of vaccination, and receipt of provider recommendation for vaccination. RESULTS: Influenza vaccine coverage was 67.8%, and PPV coverage was 60%. Coverage among blacks and Hispanics was > or = 15 percentage points below that of whites. Half (52%) of persons who had not received PPV were aware it was recommended for persons their age, and < 10% had received a recent physician recommendation for PPV. Concern about side effects and not thinking that they needed the vaccine were the most frequently cited reasons for not receiving an influenza vaccination. In each racial/ethnic group, prevalence of potential missed opportunities (recent doctor visit, but no vaccine recommendation from provider and no influenza vaccination) was higher than prevalence of potential vaccine refusal (recent doctor visit and vaccine recommendation from provider, but no vaccine): blacks, 26.9% versus 7.9%; Hispanics, 19.9% versus 12.1%; and white non Hispanics, 16.2% versus 6.1%. CONCLUSIONS: Improved adherence to vaccination guidelines by healthcare providers could substantially raise coverage in all racial/ethnic groups. Multiple factors contribute to racial/ethnic disparities, and their relative contributions should be further quantified. PMID- 16376705 TI - Immunization coverage among Hispanic ancestry, 2003 National Immunization Survey. AB - BACKGROUND: The Hispanic population is increasing and heterogeneous (Hispanic refers to persons of Spanish, Hispanic, or Latino descent). The objective was to examine immunization rates among Hispanic ancestry for the 4:3:1:3:3 series (> or = 4 doses diphtheria, tetanus toxoids, and pertussis vaccine; > or = 3 doses poliovirus vaccine; > or = 1 doses measles-containing vaccine; > or = 3 doses Haemophilus influenzae type b vaccine; and > or = 3 doses hepatitis B vaccine). METHODS: The National Immunization Survey measures immunization coverage among 19 to 35-month-old U.S. children. Coverage was compared from combined 2001-2003 data among Hispanics and non-Hispanic whites using t-tests, and among Hispanic ancestry using a chi-square test. Hispanics were categorized as Mexican, Mexican American, Central American, South American, Puerto Rican, Cuban, Spanish Caribbean (primarily Dominican Republic), other, and multiple ancestry. RESULTS: Children of Hispanic ancestry increased from 21% in 1999 to 25% in 2003. These Hispanic children were less well immunized than non-Hispanic whites (77.0%, +/ 2.1% [95% confidence interval] compared to 82.5%, +/-1.1% (95% CI) > in 2003). Immunization coverage did not vary significantly among Hispanics of varying ancestries (p=0.26); however, there was substantial geographic variability. In some areas, immunization coverage among Hispanics was significantly higher than non-Hispanic whites. CONCLUSIONS: Hispanic children were less well immunized than non-Hispanic whites; however, coverage varied notably by geographic area. Although a chi-square test found no significant differences in coverage among Hispanic ancestries, the range of coverage, 79.2%, +/-5.1% for Cuban Americans to 72.1%, +/-2.4% for Mexican descent, may suggest a need for improved and more localized monitoring among Hispanic communities. PMID- 16376706 TI - Depression as a predictor of hospitalization due to coronary heart disease. AB - BACKGROUND: Studies have shown that patients with depression have higher rates of coronary heart disease (CHD) than people in the general population. However, large-scale population-based data on incidence rates of CHD in people with depression are needed. This study analyzed whether hospitalization for depression predicts CHD in men and women after accounting for socioeconomic status and geographic region. METHODS: Data from the family coronary heart disease database at the Karolinska Institute, Stockholm, were used to identify all people in Sweden aged 25 to 64 at onset of depression and aged 25 to 79 at onset of nonfatal CHD during the study period (1987 to 2001). Standardized incidence ratios (SIRs) of CHD among those with and without depression were compared. All analyses were conducted in 2005. RESULTS: There were 1767 cases of CHD among those with depression during the study period. The risk of developing CHD was strongest for those aged <40; the SIR was 2.17 (95% confidence interval [CI]=1.50 3.03). The risk was attenuated with increasing age in both men and women. People aged 70 to 79 at onset of depression did not have an increased risk of CHD. CONCLUSIONS: Even after accounting for socioeconomic status and geographic region, depression is a clinically significant risk factor for developing CHD, especially in men and women aged 25 to 50. Primary healthcare teams should make particular efforts to identify young to middle-aged women and men who have depression, especially in combination with other CHD risk factors. PMID- 16376707 TI - Natural language processing in the electronic medical record: assessing clinician adherence to tobacco treatment guidelines. AB - BACKGROUND: Comprehensively assessing care quality with electronic medical records (EMRs) is not currently possible because much data reside in clinicians' free-text notes. METHODS: We evaluated the accuracy of MediClass, an automated, rule-based classifier of the EMR that incorporates natural language processing, in assessing whether clinicians: (1) asked if the patient smoked; (2) advised them to stop; (3) assessed their readiness to quit; (4) assisted them in quitting by providing information or medications; and (5) arranged for appropriate follow up care (i.e., the 5A's of smoking-cessation care). DESIGN: We analyzed 125 medical records of known smokers at each of four HMOs in 2003 and 2004. One trained abstractor at each HMO manually coded all 500 records according to whether or not each of the 5A's of smoking cessation care was addressed during routine outpatient visits. MEASUREMENTS: For each patient's record, we compared the presence or absence of each of the 5A's as assessed by each human coder and by MediClass. We measured the chance-corrected agreement between the human raters and MediClass using the kappa statistic. RESULTS: For "ask" and "assist," agreement among human coders was indistinguishable from agreement between humans and MediClass (p>0.05). For "assess" and "advise," the human coders agreed more with each other than they did with MediClass (p<0.01); however, MediClass performance was sufficient to assess quality in these areas. The frequency of "arrange" was too low to be analyzed. CONCLUSIONS: MediClass performance appears adequate to replace human coders of the 5A's of smoking-cessation care, allowing for automated assessment of clinician adherence to one of the most important, evidence-based guidelines in preventive health care. PMID- 16376708 TI - Death and injury from motor vehicle crashes: a tale of two countries. AB - OBJECTIVE: To determine why road deaths dropped by 33.9% in the United Kingdom, compared to 6.5% in the United States, between 1990 and 1999. METHODS: Deaths per billion vehicle kilometers traveled (D/BVKM), and case fatality rates (CFR) in the United States and the United Kingdom were tracked. Time trends in CFR can be used to track the direct effects of speed of impact. CFR is a crash-phase outcome that is independent of exposure, and varies approximately to the fourth power of the speed of crash impact. Joinpoint regression analysis was used to analyze changes in time trends of CFR. RESULTS: In the 1990s, the decrease in deaths in the United Kingdom was attributable mostly to the 29.6% drop in the CFR. In the United States, the CFR dropped by only 6.6%. The United Kingdom introduced speed cameras and an array of speed-calming measures. By contrast, in the United States, use of speed cameras was extremely rare, and speed limits and speeds increased in 32 of the 50 states, mostly in 1995 and 1996, after which CFR actually rose (p<.0001). Intercountry differences in time trends in seat belt use, trauma care, vehicle kilometers traveled, congestion, and driving under the influence of alcohol (DUI), along with massive increase in use of higher-risk sports utility vehicles in the United States, did not account for the contrasting trends in deaths through the 1990s. But increases in DUI in the United States after 1997 may have contributed to increases in speed-related crashes. CONCLUSIONS: The reductions in CFR, probably from small drops in speed of impact account for the disproportionately greater drop in death tolls in the United Kingdom compared to the United States. The temporal fit between drops in CFR and deaths following the introduction of speed cameras in the United Kingdom and increases in speed (speed creep), CFR, and deaths in the United States following raised speed limits suggests that diverging changes in speeds of impact accounted mainly for these changes. Use of D/BVKM to correct for exposure concealed the lack of progress after 1990 in the United States, since falling time trends in D/BVKM reflect increases in congestion. If the United States had implemented United Kingdom-type speed control policies and not raised speed limits, there would have been an estimated 6500 to 10,000 (approximately 16% to 25%) fewer road deaths per year during the period following speed-limit increases (1996 to 1999), including many DUI-related deaths. Reductions of up to 50% are now achievable based on newer population-wide strategies for speed control. PMID- 16376709 TI - Statewide impact of pharmacist-delivered adult influenza vaccinations. AB - BACKGROUND: Oregon law has allowed pharmacists to provide adult immunizations since 2000. Every vaccination delivered must be reported to the state health department. Previous reports indicate that pharmacists vaccinate individuals unlikely to receive vaccinations elsewhere. METHODS: Administration reports were analyzed in 2005 for the first three influenza seasons (2000 to 2003). The number of pharmacies participating, type and quantity of vaccinations, and county where provided were analyzed. RESULTS: A total of 13,116 adult patients received influenza vaccinations during 2000-2001 at 56 pharmacies. The number of pharmacies participating increased to 88 and 132, and vaccinations provided to 25,785 and 30,218 in the next two seasons, respectively. The mean number of vaccinations per pharmacy was 250 (standard deviation 236) for the 3-year period. Rural counties accounted for 28.4% of influenza vaccinations. CONCLUSIONS: Pharmacists provided a substantial number of influenza vaccinations during this 3 year period. More than one quarter of the vaccinations were provided in rural counties. PMID- 16376710 TI - Development of public health priorities for end-of-life initiatives. AB - OBJECTIVE: Recently, end-of-life (EOL) issues have captured the attention of the public health community. This study reports a project to help state health departments better understand their potential role in addressing EOL issues and develop initial priorities for EOL activities. METHODS: The project involved two studies. Study 1 (October 2002 to September 2003) involved a concept mapping process to solicit and organize recommendations from key stakeholders. Concept mapping integrates qualitative group processes with multivariate statistical analysis to represent the ideas of stakeholders visually through maps. A key informant approach was used to identify stakeholder participants with expertise in aging, cancer, public health, and EOL. In two meetings, stakeholders used the maps to develop short-, intermediate-, and long-term recommendations for EOL initiatives. Study 2 (October 2003 to September 2004) involved a modified Delphi process with three iterations to prioritize recommendations for initial action from among a group of short-term recommendations. RESULTS: Study 1 resulted in 103 recommendations for EOL initiatives across nine domains. Study 2 resulted in consensus on five initial recommendations from three domains: identifying an EOL point of contact in state health departments, collecting and analyzing data about EOL, incorporating EOL principles into state comprehensive cancer control plans, educating the public about hospice and palliative care, and educating the public about the importance of advance directives. CONCLUSIONS: Diverse perspectives of key public health stakeholders resulted in a series of short- and longer-term recommendations for EOL action. These recommendations can guide future efforts by state health departments and other public health agencies to address EOL issues. PMID- 16376711 TI - Transforming work breaks to promote health. AB - Public health, medicine, and nursing are disciplines charged with promoting health, preventing and treating disease, and improving quality of life. One of the most effective ways to achieve these objectives is to make health-promoting behaviors a part of people's everyday routines. Because many adults spend a substantial portion of the day in the workplace, and because it is common for workers to have two 10- to 15-minute work breaks each day, work breaks are a potentially important--but currently underutilized--venue in which to promote healthy behaviors. Unfortunately, work breaks frequently involve health compromising behaviors. This paper presents an argument to restructure work breaks into "booster breaks" that improve public health by encouraging health promoting practices. This new way of thinking about work breaks is described. The implications and challenges of this paradigm shift for workplace culture and public health are addressed. PMID- 16376712 TI - Health professional training in youth violence prevention: a commentary by the Surgeon General. PMID- 16376713 TI - Training health professionals in youth violence prevention: overview of extant efforts. PMID- 16376714 TI - Healthcare professionals and the prevention of youth violence: where do we go from here? PMID- 16376715 TI - The matrix comes to youth violence prevention: a strengths-based, ecologic, and developmental framework. PMID- 16376716 TI - What health professionals should know: core competencies for effective practice in youth violence prevention. AB - BACKGROUND: Youth violence has been identified as a critical health concern in the United States; however, few training resources are available for preparing health professionals to contribute to prevention efforts in their professional practices. Identification of core competencies for health professionals in youth violence prevention can be used to support the development of training resources in this area of professional practice. METHODS: In 2001, experts in youth violence, health care, and health professional education from eight of the ten Academic Centers of Excellence on Youth Violence Prevention met to develop a list of core competencies that health professionals need for effective practice in youth violence prevention. Experts participated in a 2-day facilitated session to identify these competencies. RESULTS: The group identified 40 core competencies that health professionals should acquire for effective practice in youth violence prevention. The competencies were organized across seven domains of practice and at three levels of expertise. CONCLUSIONS: Training is needed to prepare health and public health professionals to contribute to efforts in youth violence prevention in the United States. The core competencies identified by the Academic Centers of Excellence Working Group can support the development of curricula in this area. PMID- 16376717 TI - Training healthcare professionals in youth violence prevention: an overview. AB - Youth violence prevention (YVP) is a top priority for all healthcare providers. Violence ranks among the leading causes of death for children and adolescents. The Centers for Disease Control and Prevention established the Academic Centers of Excellence on Youth Violence Prevention (ACE YVP) to address this important national problem. One of the tasks of each ACE YVP is to develop YVP curricula for healthcare professionals. This article describes the experience of three ACE YVPs in developing curricula for nurses, allied health professionals, and physicians, including the identification of national resources, incorporation with local community resources, and areas of commonality and difference for incorporation at other sites. Each of the ACE YVP curricula described uses a multidisciplinary approach to teach learners. Two of the sites developed separate courses for students (University of California, San Diego and University of Puerto Rico) whereas the third integrated teaching through the medical school (University of Hawaii). PMID- 16376718 TI - Youth violence prevention in a problem-based clerkship curriculum. AB - This article describes the development of a problem-based learning case focused on youth violence prevention. Development of the case involved: a review of the literature on youth violence prevention and youth violence prevention education, drafting of a fictitious case that integrated prominent themes in the literature, review of the case by a multidisciplinary curriculum planning group whose intent was to oversee youth violence prevention training throughout medical training, and integration of the case in the psychiatry clerkship experienced by all students at the University of Hawaii John A. Burns School of Medicine. The text of the case, along with references, tutor's notes, intended learning objectives, and evaluation measures, are included in this article. Students and faculty thus far have given positive feedback on the case. We recommend that other programs consider problem-based learning as one of several methods to teach youth violence prevention and that educational outcomes be studied further. PMID- 16376719 TI - Youth violence prevention curriculum for undergraduate nursing and allied health students. AB - This paper presents the curriculum-development process, implementation, and relevant evaluation outcomes of the Introduction to Violence Prevention in Children and Youth (INTD 4065) course designed by the Center for Hispanic Youth Violence Prevention (CHYVP) of the University of Puerto Rico. The training curriculum was implemented in the academic year 2003-2004 as an undergraduate interdisciplinary three-credit course for nursing and allied health students at one of the schools of the Medical Sciences Campus, University of Puerto Rico. Objectives for the curriculum include developing competencies in child and youth violence prevention in undergraduate allied health and nursing students and promoting involvement of undergraduate health professionals in child and youth violence prevention efforts. PMID- 16376720 TI - American Academy of Pediatrics' Connected Kids program: case study. PMID- 16376721 TI - Training emergency medicine nurses and physicians in youth violence prevention. AB - Adolescents seen in an urban Emergency Department (ED) are more likely to die from violence than from any other illness or condition for which they seek care in the ED. Most injured patients presenting to our nation's EDs are treated and released, even after a firearm-related injury. These youth who are discharged from the ED will not interface with resources on the inpatient trauma unit. The current standard of care in the ED involves no referral for violence-related prevention services. Despite the fact that ED physicians and nurses frequently medically manage victims of violent assault, there are few courses on youth violence prevention (YVP) framed from the viewpoint of emergency healthcare providers, and ED staff remain relatively uneducated as a specialty on the identification, assessment, and referral resources available for early intervention and prevention. This article focuses on the development and in-depth description of a case-based, 1-hour continuing medical education presentation for ED physicians, residents, and nursing staff on YVP. This presentation is aimed to increase awareness of the role of ED personnel in YVP and to provide basic knowledge and skills needed to begin to incorporate YVP into routine clinical practice in an Emergency Department setting. PMID- 16376722 TI - American Medical Association's Youth Violence Prevention Training and Outreach Guide. AB - This article describes the development and contents of a training and outreach guide Connecting the Dots to Prevent Youth Violence: A Training and Outreach Guide for Physicians and Other Health Professionals (the Guide) on youth violence prevention for healthcare providers developed by the American Medical Association. The Guide, was developed to help translate recommendations made by the Commission for the Prevention of Youth Violence in their 2000 report, Youth and Violence: Medicine, Nursing, and Public Health: Connecting the Dots to Prevent Violence, into healthcare practice. The Guide, which will also be available in Spanish in early 2006, is structured as a speaker's kit and includes prepared speeches, case studies, issue briefs, and copies of screening tools and patient education materials from a variety of sources appropriate for use in the clinical setting. Results of a preliminary evaluation of the Guide indicate that the training can be effective in increasing providers' awareness about the problem of youth violence and encouraging them to incorporate into healthcare visits violence prevention activities such as screening youth for exposure to violence and educating patients and caregivers on strategies for reducing the risk for violence. PMID- 16376723 TI - Bringing violence prevention into the clinic: the Massachusetts Medical Society Violence Prevention Project. AB - This case study describes the experience of the Massachusetts Medical Society Violence Prevention Program. Beginning in 1996, this program developed materials to support physicians who wanted to incorporate violence prevention into primary care and urgent care practice. With technical assistance from the Harvard Youth Violence Prevention Center, the society produced parent information cards in English and Spanish, a clinical guide, and online training resources. Orders have been received from around the country and from community-based organizations and local governments. The success of this project illustrates the general interest in authoritative and useable violence prevention information. PMID- 16376724 TI - Public health training on the prevention of youth violence and suicide: an overview. AB - Although injury is the leading cause of death for Americans aged 40 and under, curricula in U.S. Schools of Public Health rarely include training on injury prevention or control. Domestically and internationally, when the topic of injury is addressed, the focus is often on unintentional injuries. Yet intentional injuries from violence and self-harm (apart from acts of war and terrorism) and the acute and chronic health problems associated with them take a large and often hidden toll on individuals, families, and communities worldwide. Adequate education on the prevention of violence and suicide by teenagers remains missing from public health and medical training. Public health and medical practitioners are confronted by violence-related injury but are provided little formal education on youth violence or suicide, effective responses, or prevention. Adolescents' involvement in violence remains a serious public health problem. Involvement in aggression and self-harm by adolescents leaves them at immediate risk of injury and often has ongoing and negative effects on future development, involvement in community and family life, and risk of morbidity and mortality for self and others. Public health practitioners are at the nexus of health care and service provision at local, state, federal, and multinational levels, and are well suited to provide training and technical assistance on youth violence prevention across disciplines and settings. In this article, training resources, opportunities, and strategies for prevention of the high prevalence of youth violence and suicide in the U.S. are discussed and recommendations for a new public health training initiative are outlined. PMID- 16376725 TI - A youth violence prevention curriculum for public health students. AB - A six-module curriculum on youth violence prevention, The Epidemiology and Prevention of Intentional Injury, was developed for inclusion in a Virginia Commonwealth University Department of Epidemiology and Community Health graduate course for public health students. The content of the curriculum was determined by reviewing key literature and the consensus group reports of experts in the area of youth violence prevention and prevention research more generally. A rationale of course content related to youth violence prevention and an overview of course content, objectives, required reading, and assignments are provided. Suggestions are made for possible adaptation of this youth violence prevention curriculum into public health courses other than intentional injury. PMID- 16376726 TI - Public health training online: the National Center for Suicide Prevention Training. AB - BACKGROUND: Suicide is a serious public health problem, and training in suicide prevention has not kept pace with recent rapid growth of the field. METHODS: To address this concern, the Harvard Injury Control Research Center and Education Development Center, Inc., launched the National Center for Suicide Prevention Training (NCSPT) with funding by the Maternal and Child Health Bureau of the Health Resources Services Administration. NCSPT offers an online professional development workshop series for public officials, service providers, and community-based coalitions involved in suicide prevention. RESULTS: Using a public health framework of prevention, the three workshops implemented to date have drawn over 1200 participants nationally and internationally over the past 3 years. Workshop participants completing the post-test and evaluation show consistent improvement in their knowledge of suicide, rate their online training experience positively, and report that they would take additional online courses if offered. A barrier to objective evaluation of the courses, however, is the high attrition rates of the courses, an expected feature of free online courses. CONCLUSIONS: Online training is a valuable option to help meet suicide prevention training needs employing flexible, easy-to-use, and inexpensive Internet technology. With its growing presence in the field, NCSPT will continue to develop new courses to improve the ability of professionals and community-based coalitions to reduce suicide and its devastating impact on public health. PMID- 16376727 TI - PREVENT: a program of the National Training Initiative on Injury and Violence Prevention. AB - Training practitioners to use evidence-based approaches to the primary prevention of violence is challenging as a result of the dearth of well-evaluated intervention programs and the lack of familiarity of some practitioners in drawing critically on existing literature. An element of the National Training Initiative in Injury and Violence Prevention, the PREVENT (Preventing Violence Through Education, Networking, and Technical Assistance) program began in late 2003 to train practitioners to address multiple types of violence by encouraging more widespread use of evidence-based approaches to primary prevention. It is intended to reach practitioners involved in addressing violence against women, sexual violence, child maltreatment, youth violence, and suicide in varied community settings. The program uses a combination of varied types of face-to face training and distance learning coupled with opportunities for networking and technical assistance. Ultimately the program intends to stimulate and facilitate changes in individual, organizational, and cultural awareness and practices fostering primary prevention of violence. The project employs formative, process, and impact evaluation techniques aimed at improving delivery of the training as well as tracking changes in individual and organizations. PMID- 16376728 TI - World Health Organization's TEACH-VIP: contributing to capacity building for youth violence prevention. AB - Youth violence is a major public health problem in every region of the world, yet it is especially prevalent in specific settings. Youth homicide rates exceeding 10.0/100,000 occur most often in countries that are low or middle income, or which are experiencing rapid economic or social change. Particularly in low- and middle-income countries, the capacity to develop and implement the comprehensive, multisectoral strategies to prevent youth violence is only just emerging. The prevention of youth violence requires multidisciplinary approaches and a variety of trained professionals. A public health approach to training in the area of injury prevention focuses on providing professionals and paraprofessionals a common understanding of essential skills and knowledge. One important benefit of this is that it addresses a major gap in current public health training that until recently has devoted relatively little attention to injury prevention. Another benefit is that it allows professionals from a variety of backgrounds to work together more effectively to reduce injury. This article will provide a broad overview of youth violence in low- and middle-income countries and will discuss the existing level of capacity within healthcare and public health sectors for responding to these problems. It concludes with a discussion of next steps for increasing capacity and a profile of the World Health Organization (WHO) training curriculum on injury and violence prevention called TEACH-VIP, an acronym for Training, Education, and Advancing Collaboration in Health on Violence and Injury Prevention, as one important effort undertaken by WHO and global injury partners to build capacity. PMID- 16376729 TI - Medical training in child maltreatment. AB - Training of medical professionals about child maltreatment may provide useful insights into the means of incorporating violence detection and prevention into healthcare practice. Despite major progress since Caffey and Kempe identified child abuse as a medical issue, more needs to be done to ensure that proper recognition, diagnostic, and reporting strategies are used when faced with the possible abuse and neglect of children. Systematic data concerning training programs are lacking. The perceived needs include more consistent education for medical professionals, more federal support for clinical research funding, higher reimbursement rates from third-party payers for clinical care for maltreated children, board certification for doctors who want to specialize in this field, and medical licensure requirements for continuing education in child and family violence. It is clear that interpersonal violence, whether it is child maltreatment, intimate partner violence, or elder abuse, is a significant public health problem in the United States, requiring support for the education and training of medical professionals. PMID- 16376730 TI - Training clinicians to identify the hidden victims: children and adolescents who witness violence. AB - In this article, we will provide an overview of the effects of witnessing violence on children and adolescents, discuss the importance of health clinicians inquiring about violence in the lives of children, and discuss the role of the clinician in educating parents about children's responses to violence. In addition, we will describe training resources that improve clinicians' skills at identifying and responding to children and their parents. PMID- 16376731 TI - Collaborating for violence prevention: training health professionals to work with schools. AB - The issue of youth violence continues to be a pressing public health problem. Increasingly schools are adopting a public health framework to address this problem and could benefit from the expertise of health professionals. The article explores opportunities for collaboration with schools, recommendations for successful collaboration, and implications for training. PMID- 16376732 TI - Homicide survivors: research and practice implications. AB - Approximately 16.4 million people in the United States have been affected by homicide. Five million adults have experienced the murder of an immediate family member; 6.6 million people have experienced the murder of a relative other than a family member, and 4.8 million have experienced the murder of a close friend. These homicide survivors experience a variety of difficulties, some similar to post-traumatic stress disorder (PTSD). The large incidence of homicide in the U.S. warrants an examination of the research on the impact of a murder on a victim's friends and family and the implications for healthcare providers. Homicide survivors experience negative psychological and physical effects that often result in an increase in the usage of primary care services. Provider training should include protocols to screen for, discuss, and make referrals for the family and friends of homicide victims. This article recommends the development of a training program to equip providers with the tools to recognize and serve this growing population of patients. PMID- 16376733 TI - Training healthcare professionals in youth violence prevention: closing the gap. PMID- 16376734 TI - The efficacy and tolerability of long-term use of dronabinol in cancer-related anorexia: a case series. PMID- 16376735 TI - Oral transmucosal fentanyl citrate and xerostomia. PMID- 16376736 TI - Correspondence between patients' preferences and surrogates' understandings for dying and death. AB - We examined the agreement between hospice patients' preferences for desired experiences during the last week of life and their surrogates' understandings of those preferences (n=92 pairs). Analyses included percent agreement, intraclass correlation coefficients, and Bland-Altman plots. Demographic characteristics and communication measures associated with better agreement were identified using t tests and analysis of variance. The median number of items on which patients and family members agreed was 14 of 30 (interquartile range, IQR 10, 16). Preferences with good agreement included both observable and non-observable experiences. Patients who reported having had conversations about treatment preferences and who reported that their surrogates knew their preferences reported higher agreement. Surrogates display a better understanding of what is important to patients at the end of life if they have had discussions about patient preferences. These discussions may enable surrogates and clinicians to more accurately follow patient preferences. PMID- 16376737 TI - Concordance of preferences for end-of-life care between terminally ill cancer patients and their family caregivers in Taiwan. AB - There is a dearth of information in the literature about the concordance of preferences for end-of-life care between terminally ill patients and their family surrogates outside the Western countries. The purpose of this study was to examine the extent of concordance in preferences for end-of-life care goals and life-sustaining treatments between Taiwanese terminally ill cancer patients and their primary family caregivers. A total of 617 dyads of patients-family caregivers across 21 hospitals throughout Taiwan were surveyed. Overall agreements on the goals for end-of-life care and preferences for initiating life sustaining treatments ranged from 62.4% to 96.9% (average: 71.0%). Kappa values for the extent of concordance ranged from 0.13 to 0.46 (average: 0.29), indicating poor to moderate consistency in personal preferences. Family caregivers had a significantly more aggressive attitude toward each examined life sustaining treatment for their ill family members than the patients' own stated preferences. In societies, such as in Asian countries, where physicians' respect for patient autonomy is frequently subordinate to the power of family, disagreements between a patient and family about end-of-life care may result in the patient's preferences being overridden at the end of life. To effect real change and to gain increased agreement on preferences for end-of-life care, an open dialogue between patients and their primary family caregivers should become standard. PMID- 16376738 TI - Measuring pain intensity in nursing home residents. AB - Assessing pain intensity in nursing home residents remains a challenge. As part of a multifaceted intervention study to improve pain practices in nursing homes, quarterly pain assessments were conducted in 12 Colorado nursing homes. Residents who reported pain or discomfort of any kind in the past 24 hours were asked to choose one of three pain intensity scales to quantify their current and highest level of pain intensity. They were also observed for pain behaviors using Feldt's Checklist of Nonverbal Pain Indicators. Residents preferred the Verbal Descriptor Scale almost 2:1 over the 11-point Verbal Numeric Rating Scale and the Faces Pain Scale. Sex and ethnicity were associated with differences in scale preference. More than one-half of residents reporting pain had an observable pain indicator. There was a monotonic relationship between reported pain intensity and number of observed pain indicators. To improve pain assessment and management in nursing homes, residents should be given a choice of pain intensity scales and observed for possible pain behaviors. PMID- 16376739 TI - Factors predicting home death for terminally ill cancer patients receiving hospital-based home care: the Lyon comprehensive cancer center experience. AB - This study aimed to determine factors favoring home death for cancer patients in a context of coordinated home care. A retrospective study was conducted among patients followed up by the home care coordinating unit of the cancer center of Lyon. The main endpoint was place of death. Univariate analysis included general characteristics (age, gender, rural or urban residence, disease), Karnofsky Index (KI), type of care at referral (chemotherapy, palliative care, or other supportive care), and coordinating medical oncologist (MCO) home visits. Significant factors were used in a logistic regression analysis. Of 250 patients, 90 (36%) had home death. Low KI and MCO home visit were correlated with home death (odds ratio, respectively, 2.1 and 3.1). These results indicate that health care support favors home death. A hospital-based home care unit is effective for bridging the gap between community and hospital. MCO home visits offer concrete support to health care professionals, patients, and relatives. PMID- 16376740 TI - Retrospective analysis of antibiotic use and survival in advanced cancer patients with infections. AB - Infection is common among advanced cancer patients. This study was undertaken to review the pattern of use of antibiotics and to identify potential factors that could affect outcomes after infection. The medical records of all patients with advanced cancer who were enrolled into the palliative care service of a district hospital during the period January, 2002 to July, 2002 were retrospectively reviewed for infections and the use of antibiotics. Among the eligible 87 patients, 17 did not have any infective episode and 70 had at least one infective episode and accounted for a total of 120 episodes. Sixty-eight episodes were associated with survival for >14 days, and 52 episodes were associated with survival of < or =14 days. The most frequent sites of infection were chest (n=63, 52.5%), urinary tract (n=35, 29.2%), and skin/wound (n=6, 5%). Antibiotics were prescribed for 97.5% (n=117) episodes. The use of second-line antibiotics was 16.2% (n=19). By multivariate logistic regression analysis, dyspnea [odds ratio (OR)=2.6, 95% confidence interval (CI)=1.1-6.3], antibiotic utilization pattern [empirical therapy (OR=4.8, 95% CI=1.7-13.2) vs. therapy according to antibiotic sensitivity], and route of administration [parenteral route (OR=3.3, 95% CI=1.3 8.2) vs. oral route] were identified as independent determinants affecting survival after infection. Dyspnea was possibly associated with poor prognosis during the treatment of infections in patients with advanced cancer, and antibiotic therapy according to sensitivity was associated with better prognosis. Further studies are encouraged to verify this. The bioethical principles on the use of antibiotics as a life-sustaining treatment should always be followed. PMID- 16376741 TI - Parental narratives of quality of life in children with leukemia as associated with the placement of a central venous catheter. AB - Thirty mothers of children with leukemia were interviewed about the child's and family's daily routines using a version of the Ecocultural Family Interview. Parental narratives were analyzed qualitatively and quantitatively. Four broad dimensions, encompassing 23 subthemes, were identified: child coping (alpha=0.88), child quality of life (alpha=0.72), parental coping (alpha=0.72), and parental trust in the medical care (alpha=0.73). Two objective variables were drawn from the medical charts (time from the diagnosis, time from central venous catheter [CVC] placement). Regression analyses showed that the number of days from the CVC placement (beta=0.46) and child coping (beta=0.44) significantly predicted children's quality of life, which in turn predicted parental trust in the medical care (beta=0.31). The methodological implications of our narrative approach are discussed. PMID- 16376742 TI - Gender differences in symptom experiences of patients with acute coronary syndromes. AB - To compare the symptom experiences between men and women with acute coronary syndromes (ACS), we surveyed a convenience sample of 112 subjects with a final diagnosis of ACS in four hospitals. Our study found that after adjusting for cardiac diagnosis, diabetes, and age, women were more likely than men to experience chest discomfort rather than chest pain; pain/discomfort only in areas of the body other than the chest; pain/discomfort that started first either in the arm(s) or in areas of the body other than the chest; and unexplained anxiety. Women were less likely than men to experience chest pain/discomfort, pain/discomfort in the left side of the chest, and chest pain/discomfort as the most worrisome symptom. Significant gender differences were observed in the reports of several symptoms associated with ACS. This study is the first to identify different pain/discomfort referral patterns between men and women that require further validation. PMID- 16376743 TI - Long-acting octreotide for the treatment and symptomatic relief of bowel obstruction in advanced ovarian cancer. AB - Symptoms of malignant bowel obstruction in patients with recurrent ovarian cancer lead to a poor quality of life. Sandostatin LAR Depot (LAR) is an intramuscular, monthly administered, long-acting form of octreotide. LAR's safety and utility were evaluated in a pilot study enrolling 15 advanced ovarian cancer patients with bowel dysfunction. Once safety with subcutaneous (SQ) octreotide was assessed, patients were given 30 mg LAR on Day 1 and octreotide SQ for 2 weeks. Of 13 evaluable patients, three patients had a major response to LAR treatment with reduction in bowel obstruction symptoms, two had a minor response, four had no response, and four had progressive symptoms. Three patients remained on LAR for more than 9 months. No significant toxicities were attributable to octreotide or LAR. Because three patients received nine or more monthly injections of LAR, possible direct antitumor effects of LAR or synergy with chemotherapy needs to be explored. PMID- 16376744 TI - When nothing helps: propofol as sedative and antiemetic in palliative cancer care. AB - Benzodiazepines, neuroleptics, and barbiturates are commonly used for sedation to achieve symptom control in end-of-life care. Propofol has several advantages over traditional sedating agents that would indicate its use in treatment-refractory situations. We report on the use of propofol in 35 patients. In 22 patients, propofol was used for palliative sedation when treatment with benzodiazepines had failed. The mean dose range during treatment was between 0.90 and 2.13 mg/kg/h. The effect was assessed as good or very good in 91% of the patients. Thirteen patients were treated with propofol due to intractable nausea and vomiting. The mean dose range during the infusion period was 0.67-1.01 mg/kg/h. The effect was judged as good or very good in 69% of the patients. Based on our experience, we propose clinical guidelines on the safe use of propofol in specialized palliative inpatient units. PMID- 16376746 TI - Vascular ultrasound: the next frontier for the echocardiographer. PMID- 16376747 TI - Insuring your workplace--a new member benefit. PMID- 16376748 TI - Evaluation of left anterior descending coronary artery stenosis of intermediate severity using transthoracic coronary flow reserve and dobutamine stress echocardiography. AB - BACKGROUND: The physiologic significance of left anterior descending coronary artery (LAD) stenosis of intermediate angiographic severity is of clinical importance and difficult to assess. Assessment of coronary flow reserve (CFR) by Doppler transthoracic echocardiography (TTE) is a new tool and could allow rapid, noninvasive evaluation of stenosis severity in this setting. OBJECTIVE: We sought to evaluate the value of CFR measurement determined by TTE, compared with dobutamine stress echocardiography (DSE), in the setting of LAD stenosis of intermediate angiographic severity. METHODS: A total of 51 consecutive stable patients in sinus rhythm (33 men; age 65 +/- 12 years; left ventricular ejection fraction 59 +/- 7%) with no previous anterior myocardial infarction and with an angiographic proximal LAD stenosis of intermediate severity (56 +/- 8% quantitative coronary angioplasty) were prospectively studied. Coronary flow velocity was measured in the distal part of the LAD by TTE at rest and during continuous infusion of 0.14 mg/kg/min of adenosine over 2 minutes, using a multifrequency transducer, in the modified parasternal or 3-apical view. CFR was calculated as the ratio of hyperemic to basal mean (mean CFR) and peak (peak CFR) diastolic flow velocity. DSE was performed immediately after the adenosine test to assess ischemia in the LAD territory (percent maximum predicted heart rate = 94 +/- 8). RESULTS: Adequate recording of CFR was possible in 46 patients. Of the 35 patients with a CFR of 2 or more (peak CFR = 2.7 +/- 0.6), DSE was normal in 34. Of the 11 patients with a CFR less than 2 (peak CFR = 1.7 +/- 0.2), 7 had an abnormal response with DSE in the LAD territory. In this range of intermediate stenosis, there was a poor correlation between percent LAD diameter stenosis and CFR. For patients with positive DSE, CFR was 1.6 +/- 0.2 compared with 2.7 +/- 0.6 for patients with normal DSE (P < .05). The sensitivity, specificity, and the positive and negative predictive values of TTE CFR for detecting ischemia on DSE were 88%, 89%, 64%, and 97%, respectively, with an overall agreement of 89% between the two tests. CONCLUSION: Given its high negative predictive value, noninvasive CFR could be a useful aid in reaching clinical decisions promptly at the bedside in patients with moderately severe lesions of the proximal LAD. PMID- 16376749 TI - Isovolumic but not ejection phase Doppler tissue indices detect left ventricular dysfunction caused by coronary stenosis. AB - BACKGROUND: Isovolumic acceleration (IVA) obtained by tissue Doppler echocardiography (TDE) is a sensitive and relatively load-independent index for assessing systolic ventricular function. IVA also has the ability to describe the force-frequency relationship during incremental atrial pacing in vivo. OBJECTIVE: We sought to assess the ability of IVA to detect global left ventricular (LV) dysfunction induced by coronary constriction. METHODS: In 6 open-chest anesthetized pigs we examined right ventricular and LV long-axis function by TDE (4-chamber view) with simultaneous invasive measurements of intraventricular pressure, maximum dP/dt, minimum dP/dt, and tau by microtip catheter. A pneumatic cuff was placed around the proximal portion of left anterior descending coronary artery (LAD) and distal flow was monitored by transonic flow probe. Mean arterial pressures were monitored by indwelling cannula. Baseline studies assessed force frequency relationships with TDE and invasive measurements during incremental pacing from 100 to 200/min (20/min increments every 10 minutes). The protocol was repeated 10 minutes after balloon inflation to reduce LAD blood flow by 50%. RESULTS: Compared with baseline, LV pressure decreased significantly (P = .03, 2 way analysis of variance) as did maximum dP/dt (P < .004) with LAD constriction. At the same time IVA and isovolumic velocity at the LV free wall were significantly reduced (P < .002 and P = .04, respectively) and both IVA and isovolumic velocity were correlated with dP/dt (r = 0.45, P < .002, and r = 0.35, P < .02, respectively). TDE systolic indices were unchanged in the right ventricle. CONCLUSION: IVA detects changes in global LV systolic function during LAD constriction and may be a useful clinical tool to diagnose ischemia. PMID- 16376750 TI - Two-dimensional strain--a Doppler-independent ultrasound method for quantitation of regional deformation: validation in vitro and in vivo. AB - BACKGROUND: A new 2-dimensional strain echocardiography (2DSE) method has been introduced that measures myocardial deformations by tracking localized acoustic markers. We compared strains measured in vitro and in vivo by 2DSE with those obtained by sonomicrometry. METHODS: For the in vitro study, a tissue-mimicking gelatin block was cyclically compressed and longitudinal strains obtained by 2DSE and sonomicrometry crystals. For the in vivo study, arrays of crystals were implanted into the apical anteroseptal (test region) and midposterior (control region) in 16 open-chest pigs and strains measured by 2DSE and crystals at baseline and after acute ischemia. RESULTS: In vitro, pooled data demonstrated good correlation (r = 0.99, P < .0001) and close agreement (bias +/- 2SD = 0.7 +/ 2.2%) of 2DSE and sonomicrometry. For a combination of low testing strains (5.4%) and strain rates (0.8 and 1.2 Hz), 2DSE overestimated strains by sonomicrometry. In vivo, linear regression analysis of pooled measurements demonstrated again a good correlation (r = 0.94, P < .0001) and agreement (1.1 +/ 7.5%) of 2DSE with sonomicrometry with a trend toward lower absolute values of strains by 2DSE. CONCLUSION: The 2DSE demonstrated good overall correlation and agreement with sonomicrometry for the tested in vitro and in vivo values. Some caution with 2DSE measurements is needed for combinations of low strains and strain rates. PMID- 16376751 TI - Preload and flow propagation velocity: insight into patients with uremia and different left ventricular systolic function. AB - OBJECTIVES: The aim of this study was to investigate the changes of flow propagation velocity (FPV) after regular hemodialysis (H/D) in patient who were uremic with different baseline left ventricular (LV) functions. METHODS: In all, 24 patients with uremia and abnormal LV systolic function were enrolled in this study and 30 patients with uremia and normal LV systolic performance served as the control group. All patients had in sinus rhythm and underwent a comprehensive transthoracic echocardiographic examination and mitral FPV by color M-mode echocardiography before and after H/D. RESULTS: The decline in FPV before H/D was proportional to the decline in the baseline LV function, whereas FPV after H/D varied according to the LV ejection fraction. An obvious decrement of FPV for patients with normal LV systolic function was noted, but no obvious difference was seen for patients with abnormal LV systolic function. CONCLUSION: FPV was strongly preload dependent for patients with normal LV systolic function. It was not significantly influenced by preload for patients with abnormal LV systolic function. FPV is a relatively good diastolic parameter for investigating patients with LV systolic dysfunction. PMID- 16376752 TI - Which Doppler parameters are load independent? A study in normal volunteers after blood donation. AB - OBJECTIVES: We sought to assess the effects of blood donation on different echocardiographic parameters in healthy volunteers. METHODS: A total of 101 healthy male volunteers were evaluated by echocardiography before and immediately after a 500-mL blood donation. In addition to traditional Doppler indices of left ventricular filling, Doppler tissue, color flow propagation, strain (epsilon), and epsilon rate were measured. RESULTS: There was a statistically significant decrease in mitral peak E and A values after blood donation (E wave 0.85 +/- 0.12 vs 0.79 +/- 0.14 cm/s, P = .01; A wave 0.65 +/- 0.10 vs 0.60 +/- 0.12 cm/s, P = .05). Mitral color flow propagation velocity was not affected (560 +/- 123 vs 571 +/- 132 mm/s, P = not significant). There were no significant differences in the Doppler tissue parameters of peak systolic, and early and late diastolic velocities after blood donation (Sm 13.5 +/- 4.6 vs 13.3 +/- 4.9 cm/s, P = not significant; Em 15.5 +/- 4.9 vs 15.9 +/- 5.1 cm/s, P = not significant; and Am 14.1 +/- 3.9 vs 14.1 +/- 3.5 cm/s, P = not significant, respectively). The peak systolic decreased significantly (-28 +/- 8% vs -21 +/- 4%, P = .03) whereas the peak systolic rate was not affected (1.5 +/- 0.35 vs 1.4 +/- 0.40 s(-1), P = not significant). CONCLUSION: In healthy male volunteers, preload reduction induced by a 500-mL blood donation does not affect the color propagation velocity, rate, and Doppler tissue velocities. PMID- 16376753 TI - Evaluation of left ventricular systolic function using automated angle independent motion tracking of mitral annular displacement. AB - Determination of ejection fraction (EF) in clinical practice typically involves manual tracing of endocardial borders. This method is time-intensive and highly dependent on image quality. Mitral annular displacement (MAD) has been shown to correlate well with EF. Previously, this method involved tedious analysis of M mode tracings. We developed and studied a new technique that is ultrasound beam angle-independent for automated detection of MAD, based on a tissue tracking algorithm. A regression formula was derived in a study group to predict EF from MAD measurements and tested prospectively in a separate group of patients. We found that our technique provides accurate, ultra-fast estimation of EF with lower inter- and intraobserver variability when compared with manually traced biplane EF. PMID- 16376754 TI - Left ventricle myocardial performance index derived either by conventional method or mitral annulus tissue-Doppler: a comparison study in healthy subjects and subjects with heart failure. AB - OBJECTIVES: The aims of this study were to investigate the clinical agreement between myocardial performance index (MPI) measured conventionally and by pulsed wave tissue Doppler (PW-TD) of the mitral annulus, and to test whether PW-TD MPI can accurately differentiate between healthy subjects and patients affected by congestive heart failure (CHF) with mild to moderate reduction of systolic function. BACKGROUND: Calculation of MPI using PW-TD may have advantages over conventional left ventricle inflow/outflow tract pulsed-wave Doppler (PWD) method; for example, all of the data needed for PW-TD MPI calculation can be derived from one single cardiac cycle, whereas with PWD at least two different cycles are needed. Thus, heart rate variability does not interfere with PW-TD MPI. METHODS AND RESULTS: In group A, we included 70 healthy adults with normal left ventricular ejection fraction and normal diastole, whereas for group B we studied 50 patients with CHF and left ventricular ejection fraction between 35% and 45%. MPI measured with PWD was statistically different (P < .0001) from MPI measured with PW-TD both in group A (0.33 +/- 0.09 vs 0.42 +/- 0.09) and group B (0.69 +/- 0.15 vs 0.79 +/- 0.12). Nonetheless, clinical agreement existed between the two methods in the single subject. Receiver operating characteristic curves showed very high accuracy for both methods to discriminate patients with CHF from healthy subjects; the optimal cutoff point was different and specific to each method: 0.50 for the conventional method and 0.60 for the PW-TD method. CONCLUSION: We found clinical agreement between MPI measured in the same subject with the conventional PWD method and with PW-TD. Both methods had similarly high diagnostic accuracy for CHF, but this study supports the use of a higher MPI cut point for best diagnostic accuracy when using the new PW-TD method. SUMMARY: We performed a study in healthy adults and in patients with congestive heart failure to investigate the clinical agreement between MPI measured conventionally and by PW-TD of the mitral annulus. We found mild agreement between MPI measured by the conventional method and by PW-TD. Both methods had high diagnostic accuracy for CHF. PW-TD method requires a higher MPI cut-point for best diagnostic accuracy. PMID- 16376755 TI - Doppler tissue imaging and color M-mode flow propagation velocity: are they really preload independent? AB - OBJECTIVE: This study investigated the change in new diastolic indexes in patients with uremia who undergo regular hemodialysis (H/D). METHODS: We studied 81 patients with uremia (41 men and 40 women) receiving regular H/D. All patients were in sinus rhythm before H/D. They had normal left ventricular systolic performance without regional wall-motion abnormality. Three patients were excluded because of atrial fibrillation after H/D. Patients were separated by the amount of body fluid removed during H/D procedure: 30 patients with H/D amount less than 2 kg (group 1), 33 patients with H/D amount between 2 and 3 kg (group 2), and 18 patients with H/D amount larger than 3 kg (group 3). They received complete transthoracic echocardiographic examinations. Flow propagation velocity (FPV) was measured by color M-mode echocardiography in apical 4-chamber view. Mitral annulus Doppler tissue velocities (peak systolic [Sa], early diastolic [Ea], and late diastolic [Aa]) were measured from septal, lateral, inferior, and posterior walls. All these parameters were obtained immediately before and after H/D. Paired data were compared. RESULTS: In patients with removed fluid amount less than 2 kg (group 1), the change of all diastolic parameters showed insignificant change except FPV (peak mitral E, P = .14; peak mitral A, P = .916; FPV, P = .009; septal Sa, P = .173; septal Ea, P = .295; septal Aa, P = .649). In patients with H/D amount between 2 and 3 kg, the change of all diastolic parameters showed statistically significant difference except Sa (peak mitral E, P = .001; peak mitral A, P = .001; FPV, P = .001; Sa, P = .589; Ea, P = .001; Aa, P = .015). In patients with H/D amount larger than 3 kg, Sa still showed insignificant change. Ea, Aa, and FPV showed significant difference after H/D (peak mitral E, P = .001; peak mitral A, P = .035; FPV, P = .008; septal Sa, P = .777; septal Ea, P = .014; septal Aa, P = .048). CONCLUSION: In patients with normal left ventricular systolic function, FPV was preload dependent. Diastolic phase mitral annulus Doppler tissue velocities (Ea and Aa) behaved differently according to the amount removed during H/D. They were preload independent when the amount removed during H/D was less than 2 kg. When the amount removed during H/D was larger than 2 kg, diastolic phase mitral annulus Doppler tissue velocities changed significantly. That is to say, diastolic phase mitral annulus Doppler tissue velocities were not totally preload independent. For systolic phase mitral annulus Doppler tissue velocity index (Sa), it was preload independent. PMID- 16376756 TI - Plasma triglyceride level is an independent predictor of altered left ventricular relaxation. AB - BACKGROUND: Diastolic dysfunction, manifested by impaired left ventricular (LV) relaxation, is prevalent among individuals with metabolic disorders. The objective of this study was to evaluate the extent to which plasma triglyceride (TG) levels are related to LV diastolic function. METHODS: A total of 424 subjects (age 49 +/- 12 years) had fasting plasma TG levels measured and underwent echocardiography for assessment of LV structure and function: LV ejection fraction and LV mass indexed to height (LVM/Ht(2.7)); transmitral inflow early diastolic peak velocity (E wave) and late diastolic peak velocity (A wave), and E wave to A wave ratio (E/A); deceleration time; and Doppler tissue imaging early diastolic myocardial velocity (EM), an index of LV relaxation. RESULTS: All subjects had normal LV ejection fraction, 48% had hypertension, 16% had increased LVM/Ht(2.7), 11% had type 2 diabetes mellitus, 37% were obese, and 27% had hypertriglyceridemia (TG > 150 mg/dL). Univariate analysis showed significant relationships between TG level and E/A, deceleration time, and Em (P < or = .001 for all). After adjustment for potential confounders in multivariate models (eg, age, systolic blood pressure, and LVM/Ht(2.7)), TG levels remained predictive of E/A, deceleration time, and Em (P < or = .05, <.001, and < or =.0001, respectively). Stepwise multivariate analysis showed that after age and body mass index, the TG level was the next most predictive variable of Em. CONCLUSIONS: Plasma TG levels show a strong relationship with impaired LV relaxation, an early marker of diastolic dysfunction in human beings. These findings support a hypothesis whereby elevated TG levels favor myocyte intracellular lipid accumulation, possibly leading to lipotoxic diastolic dysfunction. PMID- 16376757 TI - Dual triggering improves the accuracy of left ventricular volume measurements by contrast-enhanced real-time 3-dimensional echocardiography. AB - Real-time 3-dimensional echocardiographic continuous imaging (CIM) with contrast underestimates left ventricular (LV) volumes. We studied the effects of dual triggered (DT) acquisition on the accuracy of LV volume measurements for patients with poor acoustic windows. Real-time 3-dimensional echocardiographic imaging was performed in 20 patients during LV opacification (Definity) on the same day as cardiac magnetic resonance imaging. Both CIM and DT data were analyzed using custom software to calculate end-systolic volume (ESV) and end-diastolic volume (EDV), which were compared with the cardiac magnetic resonance reference. CIM correlated well with the cardiac magnetic resonance reference (EDV: r = 0.89; ESV: r = 0.93), but underestimated EDV and ESV by 17% and 19%, respectively. In contrast, DT resulted in higher correlation (EDV: r = 0.95; ESV: r = 0.96) and smaller biases (9% and 6%, respectively). In conclusion, because the accuracy of LV volume measurements depends on the acquisition strategy of contrast-enhanced real-time 3-dimensional echocardiographic images, the use of DT instead of the conventional CIM acquisition is recommended. PMID- 16376758 TI - Absence of clinically significant increase in pulmonary artery pressure after intravenous perflutren injection for myocardial perfusion imaging in pigs. AB - OBJECTIVE: We sought to evaluate the impact of a continuous intravenous infusion of perflutren on systemic pulmonary artery pressures at clinically relevant doses for myocardial perfusion imaging in pigs. METHODS: Five anesthetized, ventilated, open-chest pigs were administered perflutren intravenously at a rate of 0.0364 mL/kg/min over approximately 5 minutes. RESULTS: Optimal, sustained myocardial opacification was achieved in all animals. Perflutren produced transient, reversible increases in pulmonary artery pressures versus baseline: 10.6% (3.0 +/ 1.4 mm Hg; 95% confidence interval 1.7-4.2; P < .01) for systolic, 15.2% (2.5 +/ 1.4 mm Hg; 95% confidence interval 1.3-3.7; P < .05) for diastolic, and 11.6% (2.6 +/- 1.1 mm Hg; 95% confidence interval 1.68-3.65; P < .01) for mean pressures. Heart rate and systemic arterial pressures displayed nonsignificant increases during perflutren infusion compared with baseline. CONCLUSION: A continuous intravenous infusion of perflutren at a rate achieving optimal, sustained myocardial perfusion imaging in pigs induces a mild, transient, not clinically significant increase in pulmonary artery pressures without affecting heart rate or systemic arterial pressures. PMID- 16376759 TI - Quantitative echocardiographic evaluation of myocardial perfusion using interrupted contrast infusion technique: in vivo validation studies and feasibility in human beings. AB - BACKGROUND: We recently developed a new approach for contrast echocardiographic quantification of myocardial perfusion, based on brief interruptions of contrast infusion, which was designed to overcome the limitations of existing techniques. In this study, our technique was initially validated in a series of animal experiments designed to detect regional perfusion variations in vivo. Subsequently, clinical feasibility of perfusion measurements was tested. METHODS: Regional perfusion was measured transthoracically in 6 anesthetized pigs during baseline, partial left anterior descending coronary artery occlusion, and reperfusion, and validated with fluorescent microspheres. Adenosine-induced changes in perfusion were measured in 8 healthy volunteers. In both protocols, imaging was optimized during contrast infusion (Definity). Infusion was interrupted to allow contrast clearance and images were acquired during subsequent contrast inflow. Myocardial videointensity was measured over time and peak contrast inflow rate was calculated. RESULTS: In pigs, partial coronary occlusion resulted in a 47 +/- 23% decrease in peak contrast inflow rate in the left anterior descending coronary artery perfusion territory (P < .05), which was reversed during reperfusion, without concomitant decrease in other perfusion territories. These changes were in agreement with microspheres. In human beings, adenosine increased peak contrast inflow rate to 278 +/- 123% of baseline (P < .05). CONCLUSION: The interruption of contrast infusion technique is a sensitive tool for accurate quantification of myocardial perfusion, which may constitute an alternative to currently used techniques. PMID- 16376760 TI - Interrupted infusion of echocardiographic contrast as a basis for accurate measurement of myocardial perfusion: ex vivo validation and analysis procedures. AB - BACKGROUND: Echocardiographic quantification of myocardial perfusion is based on analysis of contrast replenishment after destructive high-energy ultrasound impulses (flash-echo). This technique is limited by nonuniform microbubble destruction and the dependency on exponential fitting of a small number of noisy time points. We hypothesized that brief interruptions of contrast infusion (ICI) would result in uniform contrast clearance followed by slow replenishment and, thus, would allow analysis from multiple data points without exponential fitting. METHODS: Electrocardiographic-triggered images were acquired in 14 isolated rabbit hearts (Langendorff) at 3 levels of coronary flow (baseline, 50%, and 15%) during contrast infusion (Definity) with flash-echo and with a 20-second infusion interruption. Myocardial videointensity was measured over time from flash-echo sequences, from which characteristic constant beta was calculated using an exponential fit. Peak contrast inflow rate was calculated from ICI data using analysis of local time derivatives. Computer simulations were used to investigate the effects of noise on the accuracy of peak contrast inflow rate and beta calculations. RESULTS: ICI resulted in uniform contrast clearance and baseline replenishment times of 15 to 25 cardiac cycles. Calculated peak contrast inflow rate followed the changes in coronary flow in all hearts at both levels of reduced flow (P < .05) and had a low intermeasurement variability of 7 +/- 6%. With flash-echo, contrast clearance was less uniform and baseline replenishment times were only 4 to 6 cardiac cycles. beta Decreased significantly only at 15% flow, and had intermeasurement variability of 42 +/- 33%. Computer simulations showed that measurement errors in both perfusion indices increased with noise, but beta had larger errors at higher rates of contrast inflow. CONCLUSION: ICI provides the basis for accurate and reproducible quantification of myocardial perfusion using fast and robust numeric analysis, and may constitute an alternative to the currently used techniques. PMID- 16376761 TI - Quantitative analysis of myocardial perfusion in rats by contrast echocardiography. AB - BACKGROUND: The ability to assess myocardial perfusion in small animals is important, especially to investigate models of myocardial ischemia. Myocardial perfusion is usually assessed by postmortem techniques, eliminating the possibility of follow-up. We sought to evaluate whether contrast echocardiography was able to quantify myocardial perfusion in rats. METHODS: Twenty-four rats divided in 3 groups (sham-operated, and 8 and 21 days after left anterior descending coronary artery stenosis) underwent myocardial contrast echocardiography using intermittent triggered imaging. Peak plateau intensity and slope of refilling were compared with myocardial blood flow achieved with fluorescent microspheres. RESULTS: High-quality images were easily obtained for each experiment. Close correlation was found between myocardial contrast echocardiography and myocardial blood flow, especially for measurements of peak plateau intensity x slope of refilling relative to the control area (y = 1.15 x 0.14, r = 0.86). CONCLUSION: Quantification of myocardial perfusion in rats is feasible by myocardial contrast echocardiography using intermittent triggered imaging. PMID- 16376762 TI - Improved visualization of aortic intima-media thickening with the use of perflutren lipid microspheres. AB - BACKGROUND: Transesophageal echocardiography (TEE) has been used to diagnose atherosclerotic disease for patients who present with systemic embolic events. The primary aim of this study was to assess the supplemental value of echocardiographic contrast to standard TEE in identifying the aortic intima medial thickness. METHODS: An aorta phantom was used to validate the accuracy of border delineation with and without contrast during TEE imaging. In all, 44 patients underwent TEE imaging of the thoracic aorta with a subsequent administration of a 0.3-mL bolus of perflutren lipid microspheres (Definity). Precontrast and postcontrast images were analyzed semiquantitatively for border delineation and quantitatively for intima-medial thickness and cross-sectional area. RESULTS: Wall thickness in the aortic phantom model with contrast was smaller, more accurate, and more reproducible to the true measurement. The clinical studies reflected similar results with average contrast intima-medial thickness measured at 0.15 +/- 0.08 cm and noncontrast at 0.18 +/- 0.08 cm (P < .01). The lateral wall showed the greatest improvement with a score of 1.23 increasing to an average of 1.82 (P < .001) with contrast. The medial, anterior, and posterior walls improved to an average 1.98, 1.39, and 2.0, respectively (P = .01). CONCLUSIONS: Contrast agents provide better aortic intima-media enhancement during TEE. PMID- 16376763 TI - Stress function index, a novel index for risk stratification and prognosis using stress echocardiography. AB - OBJECTIVES: The purpose of the study was to define an appropriate parameter for risk stratification and prognosis of patients undergoing stress echocardiography. BACKGROUND: Among stress echocardiography variables, peak wall-motion score index (WMSI) and ejection fraction (EF) have been shown to be independent and significant predictors of cardiovascular morbidity and mortality. Data on the impact and importance of each parameter in risk stratification and prognosis are limited. METHODS: We evaluated 1560 patients (59 +/- 13 years; 51% men) undergoing stress echocardiography (36% treadmill, 64% dobutamine). Peak WMSI was derived from the cumulative sum of 16 left ventricular segments divided by sum of visualized segments at peak stress. The ratio of peak WMSI to EF was calculated for the entire cohort. Based on this ratio and using the receiver operating characteristic curve, patients were divided into 3 groups: low- (< 1.9), intermediate- (1.9-3.1), and high- (> 3.1) risk subgroups. Follow-up (2.8 +/- 1.1 years) for confirmed myocardial infarction (n = 26) and cardiac death (n = 38) were obtained. RESULTS: Stress echocardiography effectively risk stratified patients into low- (0.7%/y), intermediate- (2.0%/y), and high- (4.4%/y) risk subgroups (P < .0001) based on the ratio of peak WMSI to EF. Cox proportional hazard model showed that risk stratification based on the ratio of peak WMSI to EF (global chi2 = 106.05; P < .0001) provided incremental value beyond that provided by risk stratification by peak WMSI (global chi2 = 79.23; P < .0001) or risk stratification by EF alone (global chi2 = 87.12; P < .0001). CONCLUSIONS: The ratio of peak WMSI to EF (stress function index) provides best incremental prognostic value and effectively risk stratifies patients into low-, intermediate , and high-risk subgroups and is better than risk stratification by either peak WMSI or EF alone. The results of stress echocardiography should routinely combine peak WMSI and EF for effective risk stratification. PMID- 16376764 TI - Doppler tissue imaging during supine and upright exercise in healthy adults. AB - Doppler tissue imaging variables were measured in 18 healthy men and women during supine bicycle stress exercise and upright exercise treadmill testing. From a color Doppler tissue imaging cineloop, tissue velocities in basal and mid segments of the standard 3 apical views during rest and peak exercise were assessed. Compared segments were adjusted for double product, and peak variables were compared using the paired t test. Peak systolic velocity and strain rate increased with both forms of exercise (P < .03). Time to compression-expansion crossover shortened in all segments with exercise and at peak exercise treadmill testing versus peak supine bicycle stress exercise (P < .03). Of all segments, 81% were analyzable for peak systolic velocity, 79% for strain rate. Doppler tissue imaging is a feasible tool for quantitatively assessing left ventricular response to supine and upright exercise. PMID- 16376765 TI - Exercise intolerance in patients with atrial fibrillation: clinical and echocardiographic determinants of exercise capacity. AB - Although exercise intolerance is a major symptom of patients with atrial fibrillation (AF), the factors limiting these patients' exercise capacity remains uncertain. This study evaluated the correlation of clinical and echocardiographic parameters with exercise capacity of patients with AF. In all, 73 patients (61 men and 12 women; mean age 61 years) with chronic AF were included in this study. Those patients with primary valvular diseases were excluded. Standard 2 dimensional and Doppler echocardiography was performed, and we averaged 10 consecutive measurements of each variable. Patients then underwent a symptom limited treadmill exercise testing. We also measured patients' plasma levels of B type natriuretic peptide before exercise testing. Of all clinical and echocardiographic parameters we assessed, age (r = -0.45, P = .006), ratio of early mitral inflow velocity to mitral annular velocity (r = -0.35, P = .032), and baseline heart rate were independent predictors of exercise capacity on multivariate regression analysis. In conclusion, patient's age, averaged ratio of early mitral inflow velocity to mitral annular velocity, and baseline heart rate provided useful information on exercise intolerance for patients with AF. Ratio of early mitral inflow velocity to mitral annular velocity, a noninvasive tool for estimating left ventricular filling pressure, may especially have important value for predicting functional capacity in this population as it has in individuals with in sinus rhythm. PMID- 16376766 TI - Predictive value of biphasic response during dipyridamole echocardiography test in the low-risk group of patients after acute myocardial infarction. AB - To investigate whether biphasic response during dipyridamole echocardiography test (DET), which represents viable but potentially ischemic myocardium in the infarcted region, affects prognosis of patients after uncomplicated acute myocardial infarction, we performed high-dose DET in 80 consecutive patients younger than 65 years with first acute myocardial infarction and preserved left ventricular function. Patients were followed up for 27 +/- 12 months for new coronary events. According to DET results all patients were classified into 4 groups: group 1 consisted of 20 patients with biphasic response; group 2 included 14 patients with sustained improvement in regional contractility; group 3 consisted of 24 patients showing no change in contractility; and group 4 included 22 patients with worsening response. Cumulative survival free of total coronary events was significantly lower in group 1 patients compared with all other groups (P < .05). By multivariate Cox analysis biphasic response was the strongest independent predictor of stable angina pectoris (odds ratio = 12.1, P = .0002), followed by hyperlipoproteinemia (odds ratio = 5.9, P = .006). On the other hand, development of acute coronary syndromes could not have been predicted by actual clinical or DET parameters. PMID- 16376767 TI - Echocardiographic predictors of left atrial appendage spontaneous echocontrast in patients with stroke and atrial fibrillation. AB - OBJECTIVE: The presence of spontaneous echocontrast (SEC) in the left atrium (LA) or LA appendage (LAA) has been associated with a higher risk of thromboembolism and cerebrovascular accidents. The purpose of this study was to define the new transesophageal echocardiographic predictors of SEC for patients with stroke and atrial fibrillation. METHODS: We studied 47 patients with stroke and atrial fibrillation who were undergoing transesophageal echocardiography for the evaluation of source of cardiac emboli. Patients were divided into two groups based on the absence (group 1) or presence (group 2) of SEC in the LAA. RESULTS: Compared with group 1, group 2 had larger LA, larger LAA minimum and maximum areas, and decreased LAA flow velocity, LAA wall velocity, LAA tissue intensity, and intensity ratio. In the stepwise discriminate analysis tissue intensity of LAA, minimum LAA area, LAA flow velocity, and LAA wall velocity were found as independent predictors of LAA SEC. LAA wall velocity had the greatest area under the receiver operating characteristic curve, indicating that the most powerful parameter for SEC is LAA wall velocity. CONCLUSION: Decreased LAA flow velocity and LA wall velocity, increased LAA size, and less negative LAA tissue intensity are associated with SEC in patients with nonvalvular atrial fibrillation. Strain and strain measurements of LAA give no more benefit. PMID- 16376768 TI - Left atrial appendage flow velocity as a quantitative surrogate parameter for thromboembolic risk: determinants and relationship to spontaneous echocontrast and thrombus formation--a transesophageal echocardiographic study in 500 patients with cerebral ischemia. AB - BACKGROUND: Hemostasis in the left atrial (LA) appendage (LAA) is an important cause in the formation of thrombi. Determination of the LAA flow velocity (LAAV) could be a quantitative parameter for estimating thromboembolic risk. The objective of this study was to: (1) determine the relationship between LAAV and qualitative parameters with elevated thromboembolic risk (thrombus/spontaneous echocontrast [SEC]); and (2) define factors that influence LAAV. METHODS: In all, 500 patients with stroke were examined consecutively by transesophageal echocardiography. In addition to measurement of the LAAV, the atrial appendage was examined for the presence of thrombi or SEC. RESULTS: LAAV differed significantly among patients with sinus rhythm (71 +/- 16 cm/s), paroxysmal atrial fibrillation (AF) and in sinus rhythm during transesophageal echocardiography (46 +/- 13 cm/s), paroxysmal AF and AF during transesophageal echocardiography (32 +/- 12 cm/s), and chronic AF (27 +/- 9 cm/s, P < .001). Independent of the rhythm, the risk of thrombus/SEC increased significantly at an LAAV less than 55 cm/s. At an LAAV 55 cm/s or more there is only a minimal risk of thrombus/SEC (negative predictive value 100% and 99%, respectively). Multivariate analysis showed that LAAV is the strongest predictor for the occurrence of thrombus/SEC (P < .0001). Further multivariate analysis showed that left ventricular ejection fraction, LA size, (paroxysmal) AF, age, and sex are independent parameters influencing LAAV. CONCLUSION: Independent of the basic rhythm, there is a close relationship between LAAV and qualitative parameters of elevated thromboembolic risk. LAAV could, therefore, be a quantitative surrogate parameter for risk stratification. It is influenced by both cardiac and extracardiac factors. PMID- 16376769 TI - Left atrial index is a predictor of exercise capacity in patients with hypertrophic cardiomyopathy. AB - BACKGROUND: Left atrial (LA) enlargement is related to diastolic dysfunction and mitral regurgitation (MR), both of which are common in patients with hypertrophic cardiomyopathy (HCM). This study investigates the association between LA size and exercise capacity in patients with HCM. METHODS: All HCM patients who underwent a treadmill test with direct measurement of oxygen consumption (VO2) and a standard transthoracic echocardiography within 30 days in the years 2001-2003 were identified. Patients with significant comorbidities were excluded. Exercise capacity was defined as percentage of predicted peak VO2. Clinical and echocardiographic parameters were compared with those of a group of normal subjects. RESULTS: Compared with normal subjects, HCM patients had increased left atrial (LA) volume index (36 vs 21 mL/m2; P < .0001) and mitral E/e' ratio (14 vs 9; P < .0001); 27% of the patients had at least moderate MR. LA volume index demonstrated borderline correlation with exercise capacity (r = -.20; P = .06) but was an independent predictor of exercise capacity in a multivariate linear analysis, together with body mass index, heart rate at rest, and left ventricular end-systolic diameter. Including the parameters E/e' ratio or moderate or severe MR did not add incremental value to the model. CONCLUSION: LA volume index, reflecting the combined influences of MR and diastolic dysfunction, was independently associated with objective measures of exercise capacity in patients with isolated HCM. PMID- 16376770 TI - Transesophageal assessment of left atrial thrombus using a 3.3-mm monoplane probe. AB - OBJECTIVES: Our study was designed to demonstrate that transesophageal echocardiography using a 3.3-mm monoplane probe can accurately evaluate the left atrium for patients with arrhythmias before cardioversion. BACKGROUND: Standard probes cause discomfort during intubation requiring sedation, but miniature probes do not. METHODS: With topical anesthesia alone, a 3.3-mm probe was used for transesophageal echocardiography in 60 patients. After intravenous sedation, standard transesophageal echocardiography was then performed. RESULTS: In 51 of 60 patients, the left atrium was visualized with the 3.3-mm probe. In 43 of 51 patients the appendage was clear. A thrombus was seen in 7 patients on both studies. In one patient spontaneous echocontrast was seen only with the 3.3-mm probe (sensitivity 100%, specificity 97%). In 9 of 60 patients, the appendage could not be assessed. CONCLUSIONS: In many patients the 3.3-mm probe can visualize the appendage and obviate the need for sedation. Technical advances will improve image quality with miniature probes. PMID- 16376771 TI - Is surgical closure of patent foramen ovale the gold standard for treating interatrial shunts? An echocardiographic follow-up study. AB - BACKGROUND: Surgical closure of patent foramen ovale (PFO) has been considered definitive and the gold standard for preventing recurrent paradoxical embolism. However, in contrast to transcatheter PFO closure, patients undergoing operation have not systematically been re-evaluated for residual shunting. This study aimed to assess the efficacy of surgical PFO closure during follow-up by transesophageal echocardiography (TEE). METHODS: Eleven adult patients with diagnosis of a PFO by contrast and/or color Doppler TEE underwent PFO closure by thoracotomy and direct suturing because of paradoxical embolism (n = 4), because of impending paradoxical embolism (n = 1), or during valve operation (n = 6). RESULTS: TEE performed 5 days to 7 months after PFO closure revealed residual shunting in 8 of 11 patients (73%). The shunt size was unchanged in two patients. According to the color Doppler jet width across the atrial septum, the postoperative PFO diameter was smaller in 5 of the remaining 6 patients. Right-to left shunting by contrast TEE, however, had newly developed (n = 1) or increased (n = 2) for patients with concomitant valve operation. Despite therapeutic anticoagulation a cerebrovascular event occurred in one patient 4 weeks after attempted PFO closure. The mechanism for persistent shunting was incomplete sealing of septum primum and septum secundum by the suture line (n = 6) or a new iatrogenic defect of the fossa ovalis caused by surgical manipulation (n = 2). In two patients a second TEE after 12 and 41 months revealed enlargement of the PFO diameter. CONCLUSIONS: Surgical PFO closure can not be regarded as the gold standard for definitive treatment of interatrial shunts. Residual shunting present in a high proportion of patients may partly explain the recurrence of embolic events. PMID- 16376772 TI - Planimetric assessment of anatomic valve area overestimates effective orifice area in bicuspid aortic stenosis. AB - BACKGROUND: Although the continuity equation remains the noninvasive standard, planimetry using transesophageal echocardiography is often used to assess valve area for patients with aortic stenosis (AS). Not uncommonly, however, anatomic valve area (AVAA) obtained by planimetry overestimates continuity-derived effective valve area (AVAE) in bicuspid AS. METHODS: Transthoracic Doppler and transesophageal echocardiography were performed to obtain AVAE and AVAA in 31 patients with bicuspid AS (age 61 +/- 11 years) and 22 patients with degenerative tricuspid AS (age 71 +/- 13 years). Aortic root and left ventricular outflow tract dimensions and the directional angle of the stenotic jet were assessed in all patients. Using these data, a computational fluid dynamics model was constructed to test the effect of these variables in determining the relationship between AVAE and AVAA. RESULTS: For patients with tricuspid AS, the correlation between AVAA (1.15 +/- 0.36 cm2) and AVAE (1.13 +/- 0.46 cm2) was excellent (r = 0.91, P < .001, Delta = 0.02 +/- 0.21 cm2). However, AVAA was significantly larger (1.19 +/- 0.35 cm2) than AVAE (0.89 +/- 0.29 cm2) in the bicuspid AS group (r = 0.71, P < .001, Delta = 0.29 +/- 0.25 cm2). Computer simulation demonstrated that the observed discrepancy related to jet eccentricity. CONCLUSION: For a given anatomic orifice, functional severity tends to be greater in bicuspid AS than in tricuspid AS. This appears to be primarily related to greater jet eccentricity and less pressure recovery. PMID- 16376773 TI - Doppler echocardiography of normal Starr-Edwards mitral prostheses: a comprehensive function assessment including continuity equation and time-velocity integral ratio. AB - The purpose of this study was to provide comprehensive Doppler echocardiographic assessment of the function of the normal Starr-Edwards mitral valve prosthesis using all the Doppler hemodynamic variables described to date, including the mitral valve prosthesis time-velocity integral (TVI)/left ventricular outflow tract TVI ratio and the prosthesis performance index. All patients had a peak early mitral diastolic velocity of no more than 2 m/s or a pressure half-time that was less than 130 milliseconds. All but one patient had either a peak early mitral diastolic velocity of no more than 2 m/s or a mitral valve prosthesis TVI/left ventricular outflow tract TVI ratio of less than 2.2, regardless of prosthesis size or left ventricular systolic function. There was a trend of decreasing prosthesis performance index with increasing prosthesis valve size that was not statistically significant, however. PMID- 16376774 TI - Predictors of mortality in paravalvular abscess. AB - BACKGROUND: Paravalvular abscess formation is an ominous complication of infective endocarditis; however, prognostic variables in paravalvular abscess are poorly defined. METHODS: We examined our experience in patients with paravalvular abscess between 1987 and 2004. Clinical, echocardiographic, microbiologic, and surgical data were examined. RESULTS: There were 45 patients (17 females), age 57 +/- 17 years. Twenty-four patients had prosthetic valve endocarditis. Methicillin sensitive Staphylococcus aureus and coagulase-negative S. aureus were the most common organisms accounting for 25 (56%) cases. Thirty-eight patients (84%) underwent surgery during initial admission. Surgical mortality was 7%, in hospital mortality was 31%, and 1-year mortality was 38%. Between patients who died and patients who survived, there were no differences in age (61 +/- 20 years vs 55 +/- 15 years, P = .3), type of microorganism, presence of prosthetic heart valves (47% vs 57%), presence of moderate to severe or severe regurgitation of involved valve (47% vs 57%, P = .37), presence of associated valvular vegetation (93% vs 93%), area of abscess (5.6 +/- 2.9 cm2 vs 4.4 +/- 3.2 cm2, P = .39), left ventricular systolic function (56% +/- 13% vs 56% +/- 10%, P = .9), white cell count (13 +/- 4 vs 13 +/- 7, P = .9), or polymorphonuclear leukocytosis (86% +/- 6% vs 81% +/- 9%, P = .1). Patients who died were sicker on admission compared with those who survived (33% had stroke or altered mental status vs 7%, P = .03) and had worse renal function compared with those who survived (creatinine 4 +/- 4 mg/dL vs 1.6 +/- 1.9 mg/dL, P = .009). CONCLUSION: Neurologic impairment and renal impairment are significant determinants of 1-year survival in patients who present with paravalvular abscess. PMID- 16376775 TI - Echocardiographic evaluation of the mitral valve area before and after percutaneous mitral commissurotomy: the pressure half-time method revisited. AB - BACKGROUND: Doppler pressure half-time (PHT) is widely used for mitral valve area (MVA) assessment but its accuracy has not been fully evaluated before and after percutaneous mitral commissurotomy (PMC) in a large series of patients. METHODS: In 120 patients with severe mitral stenosis, MVA(PHT) was prospectively evaluated before and 24 to 48 hours after PMC and compared with 2-dimensional planimetry (MVA(2D)) as a reference method. RESULTS: After PMC, MVA(2D) significantly increased (1.81 +/- 0.30 vs 1.03 +/- 0.23 cm2, P < .0001), mean transmitral gradient decreased (5 +/- 3 vs 10 +/- 5 mm Hg, P < .0001), and a good valve opening (MVA(2D) > or = 1.5 cm2) was observed in 107 patients (89%). Before PMC, correlation between MVA(PHT) and MVA(2D) was only fair overall (r = 0.52, P < .0001) and weak in subgroups of older patients (> or = 60 years; r = 0.16, P = .37) and in patients in atrial fibrillation (r = 0.38, P < .05). After PMC, MVA(PHT) (1.62 +/- 0.39 cm2) was significantly lower than MVA(2D) (P < .0001) and correlation was poor overall (r = 0.30, P = .0004; mean difference 0.33 +/- 0.30 cm2) and in all subgroups (r < 0.35). However, for the prediction of a good valve opening, a PHT less than 130 milliseconds (observed in 43 patients, 36%) had an excellent specificity (100%) despite a poor sensitivity (44%). CONCLUSION: For MVA assessment, the PHT method should be used cautiously even before PMC, especially in older patients or those in atrial fibrillation. After PMC, it does not provide an accurate MVA evaluation but can still be used as a semiquantitative method: a PHT less than 130 milliseconds is associated with a good valve opening, which can be useful in difficult cases. PMID- 16376776 TI - Noninvasive localization of nuclear factor of activated T cells c1-/- mouse embryos by ultrasound biomicroscopy-Doppler allows genotype-phenotype correlation. AB - Ultrasound biomicroscopy (UBM)-Doppler allows study of cardiovascular physiology in the in utero mouse embryo from embryonic day (E)8.25 onward. We determined the accuracy of localization of embryos by transabdominal, noninvasive 40-MHz UBM Doppler imaging. Nuclear factor of activated T cells c1-/- mice lack semilunar valves, exhibit outflow tract regurgitation, and die in utero. In timed pregnant mice generated from heterozygote crosses, an UBM-derived map of the in situ litter was compared with a definitive laparotomy map, and UBM-Doppler cardiac screen attempted for each embryo. All 109 living and dead (nonresorbed) E10.5 to 17.5 embryos were imaged and accurately localized. All 10 embryos with reversed diastolic aortic flow and 7 of 9 dead embryos genotyped were nuclear factor of activated T cells c1-/-. In 30 embryos followed up serially over 1 to 2 days from E12.5 to E16.5, we again achieved 100% accuracy in localizing at follow-up. Noninvasive localization and UBM-Doppler analysis of in situ mouse embryos can provide accurate genotype-phenotype correlation, along with nontraumatic serial imaging of embryos. PMID- 16376777 TI - Long-term follow-up of intramyocardial dissecting hematomas complicating acute myocardial infarction. AB - Intramyocardial dissecting hematoma is a form of subacute cardiac rupture complicating acute myocardial infarction. Initially contained within the myocardial wall, the hematoma may expand, rupture into adjacent structures, or spontaneously resolve. However, long-term follow-up is unknown because clinical and serial imaging data are lacking. The purpose of this study was to characterize the early and late myocardial wall changes after transmural myocardial infarction using serial ultrasound examinations of the infarct-related segments. Clinical, electrocardiographic, and echocardiographic features of 8 patients (7 men, mean age 59 years) who presented with acute myocardial infarction and echocardiographically documented intramyocardial dissecting hematoma were analyzed. All patients had precordial echocardiography and 6 underwent transesophageal echocardiography. Differentiating hematoma from trabeculations, thrombus, or pseudoaneurysm was done with contrast and color flow Doppler. Seven patients presented with S-T elevation in V1 to V4, and in 3 the elevation extended to V5, V6, I, and aVL. One patient presented with S-T elevation in II, III, aVF, V3R, and V4R. The most striking feature was persistent S-T elevation of more than 72 hours in all patients. Hematoma consisted of a cysticlike, echolucent cavity variable in size, adjacent to severely hypokinetic or dyskinetic infarct-related segments. Hematoma acoustic characteristics depended on time of evolution. Two patients underwent elective revascularization and the rest were medically treated. Two patients died and 6 were alive at the mean follow-up of 12 months. In conclusion, persistent S-T elevation is an important clue in suggesting intramyocardial dissecting hematoma, which is confirmed by its unique ultrasound appearance. Serial echocardiography is useful in determining its evolving nature, and may guide outcome. PMID- 16376778 TI - Left ventricular support by axial flow pump: the echocardiographic approach to device malfunction. AB - Axial flow pumps have gained increased acceptance in recent years as a bridge to heart transplantation and, more recently, as destination therapy. As left ventricular (LV) assist device dysfunction will be increasingly prevalent, the aim of our work was to introduce an echocardiographic management protocol as a guide to recognize the causes of pump failure. In this article we describe the echocardiographic approach to 5 episodes of malfunction of an axial flow pump (DeBakey, MicroMed Technology Inc, Houston, Tex) in 4 patients: 4 episodes caused by thrombosis of LV assist device and one caused by abnormal increase of systemic vascular resistance. In our experience, echocardiography played a pivotal role in clinical management of LV assist device failure. It allowed us to: assess patency and position of inflow and outflow cannulae; research the source of thromboembolic material; assess adequate LV filling and unloading; and optimize right ventricular function, volume replacement therapy, and pharmacologic support. PMID- 16376779 TI - Early diagnosis of stress-induced apical ballooning syndrome based on classic echocardiographic findings and correlation with cardiac catheterization. AB - Stress-induced apical ballooning has been described as a reversible condition involving the apical left ventricular wall, sparing the base, and causing a ballooning appearance of the left ventricle during systole despite normal coronaries. We are presenting 4 cases of apical ballooning seen at our institution with echocardiographic correlation. Echocardiography showed similar anatomical apical ballooning of the left ventricular apex. The diagnosis of apical ballooning syndrome was suspected based on echocardiography in conjunction with clinical data before cardiac catheterization was performed. In one case, in addition to classic left ventricular apical ballooning, marked right ventricular apical akinesia was present on the initial echocardiographic examination. This makes diagnosis of apical ballooning syndrome most likely in this patient before cardiac catheterization. Therefore, we suggest using echocardiography more often for the early diagnosis of this disease, based on careful anatomic evaluation in conjunction with clinical data. Wall motion analysis should reveal an apical ballooning appearance involving many coronary territories. Furthermore, the additional presence of right ventricular apical akinesia during echocardiographic examination makes the diagnosis of this syndrome more likely. PMID- 16376780 TI - Retained left atrial catheter: a case report and review of the literature. AB - We describe a patient in whom transesophageal echocardiography detected a portion of retained left atrial catheter that had been left in situ for 21 years after aortic valve replacement. Although thrombus was present on the catheter, the patient had no history of systemic embolization, potentially as a result of the use of chronic anticoagulation with warfarin. A review of the literature identified 8 previous cases of a retained left atrial catheter. Retained left atrial catheters can be easily detected with transesophageal echocardiography and appear to have a significant risk of thrombus formation and systemic embolization that warrants removal of the catheter when identified. PMID- 16376781 TI - Echocardiographic assessment of ventricular asynchrony in dilated cardiomyopathy and congenital heart disease: tools and hopes. AB - Ventricular dyssynchrony is a relatively common problem in patients with heart failure, in particular those with wide QRS complex, and appears to have a deleterious effect on the natural history of heart failure, as it has been associated with increased mortality. Mechanistic studies, observational evaluations, and randomized trials have consistently demonstrated the beneficial effects of cardiac resynchronization therapy (CRT) in patients with moderate-to severe chronic systolic heart failure and ventricular dyssynchrony who have failed optimal medical treatment. However, despite the promising results, it is estimated that in approximately 30% of patients undergoing CRT, the symptoms of heart failure do not improve or become even worse. One of the most important reasons for this failure is probably the lack of distinct mechanical dyssynchrony before implantation. A number of echocardiographic tools have been developed during the past 3 years for quantitative measurement of the severity of dyssynchrony before and after CRT. This review discusses the actual and potential role of different echocardiographic techniques in selection of patients and optimization of CRT and the value of some new clinical applications such as in congenital heart disease. PMID- 16376782 TI - Recommendations for chamber quantification: a report from the American Society of Echocardiography's Guidelines and Standards Committee and the Chamber Quantification Writing Group, developed in conjunction with the European Association of Echocardiography, a branch of the European Society of Cardiology. PMID- 16376783 TI - Can we finally measure blood flow velocity all through the coronary artery three by transthoracic Doppler echocardiography in patients with myocardial hypertrophy? PMID- 16376785 TI - ISAT and ISUIA: the impact on informed consent. AB - The International Subarachnoid Aneurysm Trial (ISAT) was a multicenter randomized trial that compared the safety and efficacy of endovascular coil treatment versus surgical clipping for the treatment of ruptured brain aneurysms. To be eligible for enrollment in the ISAT trial, each patient had to be deemed equally suitable for either coiling or clipping. The investigators used the term "clinical equipoise" to describe this balance. This study more than any other has set the playing field for the future of interventional radiology/endovascular neurosurgery politics. PMID- 16376786 TI - Endovascular management of a patient after SAH. AB - Treatment of acute aneurysmal subarachnoid hemorrhages consists of occluding the aneurysm to prevent rebleed, attempting to prevent vasospasm, and maintaining blood flow to the brain through vessels in vasospasm. Endovascular treatment has been shown to be as safe as, or safer, than surgical clipping for patients with SAH. Engineering solutions to our clinical problems continue to improve endovascular outcomes. This article reviews the current state of endovascular therapy. PMID- 16376787 TI - Endovascular treatment of cerebral arteriovenous malformations. AB - The modern management of cerebral arteriovenous malformations (AVMs) is based on three therapeutic modalities: microneurosurgery, endovascular embolization, and stereotactic radiosurgery. Embolization facilitates subsequent radiosurgery by reducing the volume of the nidus, prepares the resection of surgically accessible AVMs, and immediately addresses the risks related to associated intra/extranidal aneurysms and arteriovenous fistulas. We discuss in this article the current state of AVM endovascular therapy. PMID- 16376788 TI - Worksheets for use while performing carotid stenting or stroke intervention. PMID- 16376789 TI - Endovascular treatment of vertebral artery origin lesions. AB - Endovascular management of vertebral artery stenosis has been shown to have significantly better outcomes than surgical therapy or medical therapy alone. The average mortality is zero and morbidity is about 1%. There is a 97% procedural success rate. The issue is a restenosis rate that is between 30% and 41%. This article will discuss the indications, procedure, and treatment options. PMID- 16376790 TI - Embolization for intractable epistaxis. AB - It is estimated that 60% of the population experiences epistaxis at least once in a lifetime, and that approximately 6% of these patients require treatment. We discuss the risks and benefits of an endovascular approach in our experience. We believe this option is often offered too late. By following basic rules the procedure is very safe and highly effective. PMID- 16376792 TI - Molecular control of liver development. AB - Recent studies using animal models have elucidated a growing number of evolutionarily conserved genes and pathways that control liver development from the embryonic endoderm. It is increasingly clear that the genetic programs active in embryogenesis are often deregulated or reactivated in disease, cancer, and tissue repair. Understanding the molecular control of liver development should impact diagnosis and treatment of pediatric and adult liver diseases and aid in efforts to differentiate liver tissue in vitro for stem cell-based therapies. PMID- 16376793 TI - Whatever happened to "neonatal hepatitis"? AB - 'Idiopathic neonatal hepatitis' is a term that has traditionally been used to denote a clinical syndrome manifest by prolonged jaundice in the neonate. This description is now used much less frequently because recent studies unite well defined clinical, biochemical and molecular features of intrahepatic cholestasis into specific syndromes. Advances in the understanding of the molecular basis of cholestatic syndromes now enable the classification of syndromes based on biology and offer an opportunity to develop new diagnostic approaches and treatment strategies that take into account the genetic make-up of the child with cholestasis. PMID- 16376794 TI - The hepatic fibrocystic diseases. AB - The hepatic fibrocystic diseases present with variable intrahepatic biliary abnormalities, which range from portal tract enlargement and fibrosis to cystic formations. They may present as autosomal recessive or dominant polycystic kidney diseases, with associated dilatation of the renal collecting system, or as incompletely characterized cystic diseases. Symptoms from the liver disease often result from complications of fibrosis or dilated ducts/cyst (sludge, lithiasis, infection). The treatment is supportive, with careful attention to associated renal disease. Liver transplantation is an option in selected patients. PMID- 16376795 TI - Pathogenesis and treatment opportunities for biliary atresia. AB - Biliary atresia is the single most common cause of chronic cholestasis in children and is the leading indication for liver transplantation worldwide. It results from an inflammatory and fibrosing obstruction of the extrahepatic bile ducts in the first few months of life. Early diagnosis and timely surgical portoenterostomy are necessary for improved biliary drainage, but the liver disease progresses to end-stage biliary cirrhosis in most patients. Although the pathogenesis of disease is largely unknown, recent patient- and animal-based experiments indicate interactions between infectious agents and inflammatory circuits may be important pathogenic mechanisms of disease. PMID- 16376796 TI - Autoimmune hepatitis and primary sclerosing cholangitis. AB - Autoimmune liver disease in children presents predominantly as autoimmune hepatitis (AIH) and primary sclerosing cholangitis (PSC). These diagnoses must be considered in patients who have acute and chronic hepatitis, particularly when an extrahepatic autoimmune disorder is present. In AIH, the timely and sustained control of liver inflammation is critical to improve the short- and long-term outcomes. No effective treatment for PSC has been identified to date, but supportive care, careful attention to complications and associated nonhepatic diseases, and liver transplantation significantly improve the long-term outcome. PMID- 16376797 TI - Nonalcoholic fatty liver disease in children. AB - Nonalcoholic fatty liver disease (NAFLD) is likely to reach epidemic proportions in children worldwide in the next decade. NAFLD may be the hepatic aspect of the metabolic syndrome in adults and children. The entire range of liver involvement characterizing NAFLD can occur in children: hepatic macrovesicular steatosis without inflammation, steatosis with inflammation or fibrosis, and cirrhosis. NAFLD may be more severe in children from certain ethnic groups or in association with metabolic disorders characterized by abnormalities in insulin receptor structure and function. Treatment strategies focus on modifying risk factors because specific drug treatments are lacking. Overweight/obesity should be identified as early as possible. Comprehensive clinical management to normalize weight should be instituted immediately to avoid hepatic and nonhepatic complications. PMID- 16376798 TI - Hepatitis B and C in children: current treatment and future strategies. AB - A large proportion of chronic hepatitis B virus (HBV) infections are acquired during childhood. Fewer chronic hepatitis C virus (HCV) infections occur in children than in adults, but thousands of children worldwide have this serious infection. Optimal treatment strategies for these chronic infections in children have not been determined, because data on the natural history are limited. Few medications are currently approved for use in this population. This article discusses how chronic viral hepatitis differs in children and adults in epidemiology, natural history, progression of disease, and response to treatment. Treatment options are discussed, including patient selection, specific medications, ongoing studies, and future treatment options. PMID- 16376799 TI - Acute liver failure in children. AB - Acute liver failure (ALF) is a rare but devastating illness. Specific therapy to promote liver recovery is often not available, and the underlying cause of the liver failure is often unknown. This article examines current knowledge of the epidemiology, pathobiology, and treatment of ALF in children and identifies potential gaps in this knowledge for future study. PMID- 16376800 TI - Pediatric liver transplantation. AB - In the past two decades, pediatric liver transplantation has become the state-of the-art operation with anticipated success and limited mortality. The future success of pediatric liver transplantation will require thoughtful solutions to the delicate balance of risk to donors and recipients, the complex needs of the acute postoperative patient, and the long-term challenges of chronic immunosuppression in these previously unsalvageable patients. PMID- 16376801 TI - Rigorous evaluation of functional rehabilitation and physiotherapy: an imperative. PMID- 16376802 TI - Parkinson's disease and osteoporosis. AB - Parkinson's disease is associated with an increased risk of falls. The risk is greatest in patients with advanced disease. Because Parkinson's disease usually occurs late in life, the risk factors related to the neurological impairments add to those associated with aging. The incidence of fractures is high in patients with Parkinson's disease, with femoral neck fractures in older women being particularly common. Risk factors for fractures include a low body mass index, limited exposure to sunlight, an inadequate vitamin D intake with low 25-OH vitamin D levels, and bone loss. Several studies found decreased bone mineral density values at the femoral neck and lumbar spine in patients with Parkinson's disease. Although this decrease is ascribable in part to factors unrelated with Parkinson's disease, such as older age and female gender, Parkinson's disease itself also plays a role, most notably in patients with severe neurological impairments (Hoehn and Yahr stages III and IV). PMID- 16376803 TI - Pleuropulmonary involvement in ankylosing spondylitis. AB - Pleuropulmonary involvement was long described as an uncommon and late event in the course of ankylosing spondylitis (AS). This belief was based on studies that relied on symptoms and chest radiographs to evaluate the lungs. However, pleuropulmonary involvement in AS patients is usually asymptomatic, and the early lesions are undetectable on chest radiographs. Apical fibrosis, interstitial infiltrates, and pleural thickening were considered to be the main patterns. However, the introduction of high-resolution computed tomography (HRCT) has led to the description of many pulmonary abnormalities that are clinically silent and undetectable on plain radiographs. These abnormalities mainly affect the interstitium and have no influence on respiratory function, which is dependent on the severity of chest wall inflammation or ankylosis in recent-onset and established AS, respectively. Cytological and histological studies suggest that, in common with uveitis and aortic regurgitation, the structural lung changes shown by HRCT may be specific of AS. PMID- 16376804 TI - Bone and joint disease associated with primary immune deficiencies. AB - Primary immune deficiencies (PIDs) are characterized by functional and/or quantitative abnormalities of one or more immune system components. Several bone and joint abnormalities can occur in patients with PID, with arthritis being the most common. Joint manifestations, of which arthritis is the most common, occur chiefly in humoral PIDs (agammaglobulinemia, common variable immunodeficiency, hyper-IgM syndromes, and IgA deficiency) and occasionally in other PIDs (chronic granulomatous disease and Wiskott-Aldrich syndrome). Monoarthritis or oligoarthritis is the usual pattern, although polyarthritis may occur, occasionally with nodules suggesting rheumatoid arthritis. Arthritis in patients with PID is usually infectious in nature, the most common causative organism being Mycoplasma, followed by Staphylococcus, Streptococcus, and Haemophilus. These bacteria can induce not only synovial infections, but also aseptic arthritogenic inflammatory responses. Arthritis having no demonstrable relation to chronic infection has been reported also and ascribed to dysimmunity-driven mechanisms that exhibit a number of specific features. Bone lesions are far less common and usually due to infections complicating humoral PID. Distinctive bone manifestations occur in a number of rare PIDs (e.g., hyper-IgE syndrome and Di George syndrome) and in syndromes characterized by spondyloepiphyseal dysplasia. Familiarity with PID syndromes both enhances the diagnostic capabilities of physicians and provides insight into the pathophysiology of bone and joint abnormalities associated with immune dysfunction. In children and occasionally in adults, a combination of bone and/or joint manifestations and hypogammaglobulinemia may indicate PID. When there is no evidence of lymphoproliferative disease, infection, or iatrogenic complications, investigations for PID should be obtained. PID-related arthritis is a unique model for studying the pathogenesis of presumably postinfectious arthritis and of inflammatory joint diseases including rheumatoid arthritis. PMID- 16376805 TI - Magnetic resonance imaging quantification of hand synovitis in patients with rheumatoid arthritis treated with infliximab. AB - AIM: To investigate the clinical response and to evaluate by magnetic resonance imaging the inflammatory tissue changes in refractory rheumatoid arthritis patients treated with infliximab. METHODS: Sixteen refractory rheumatoid arthritis patients who were treated with intravenous infliximab (3 mg/kg) at weeks 0, 2, 6 and every 8 weeks thereafter were examined with magnetic resonance imaging of the dominant affected wrist and hand before treatment and 1 year after therapy. The volume of the enhancing inflammatory tissue was evaluated in fat suppressed contrast enhanced T1-weighted images by using the Analyse 4.0 software. Disease activity was evaluated by assessing the disease activity score for 28 joint indices. The clinical improvement was evaluated according to the American College of Rheumatology 20% response criteria. RESULTS: There were 13 females and 3 males with mean age 49.5 (17.0) years and mean disease duration 10.5 (8.0) years. Ten patients had positive IgM rheumatoid factor. One year after treatment, a significant reduction of the erythrocyte sedimentation rate, the C reactive protein, the disease activity score for 28 joint indices and the volume of the enhancing inflammatory tissue was observed. All but two of the rheumatoid arthritis patients achieved the American College of Rheumatology 20% response criteria, while 9 (56.25%) and 5 (31.25%) patients achieved the 50% and 70% American College of Rheumatology response criteria, respectively. A positive correlation among the volume of the enhancing inflammatory tissue, swollen joint count, tender joint count, as well as disease activity score for 28 joint indices (r=0.66, r=0.79, r=0.57 respectively) was found before treatment. CONCLUSIONS: In refractory rheumatoid arthritis patients, the addition of infliximab therapy may result in clinical, laboratory and magnetic resonance imaging improvement. Magnetic resonance imaging assessment of the volume of the enhancing inflammatory tissue may represent an additional tool for the investigation of joint disease activity and responsiveness to treatment. PMID- 16376806 TI - No serological evidence that fibromyalgia is linked with exposure to human parvovirus B19. PMID- 16376808 TI - Druggability: selecting optimized drug candidates. PMID- 16376809 TI - Massive variability in a stable binding protein. PMID- 16376810 TI - Nanomedicine transforms drug delivery. PMID- 16376811 TI - Sourcing natural killer cells for antitumor immunotherapy. PMID- 16376812 TI - Hope for Huntington's from an old antibiotic. PMID- 16376813 TI - Multiple sclerosis poses tough drug development challenges. PMID- 16376814 TI - Medicinal mushroom cuts off prostate cancer cells' blood supply. PMID- 16376815 TI - Andy Southan talks about ion channel research services at BioFocus. PMID- 16376816 TI - Antimalarial therapies. AB - Analysis of the trends in antimalarial research depicts a slow growth in launched products since 1995. However, 2002 saw the completion, after ten years, of the malarial genome project and an upsurge in products entering active development can be seen at the end of this period. PMID- 16376817 TI - Rapid growth in Israel's life science industry. PMID- 16376818 TI - Microarrays, databases and hard, hard sums. PMID- 16376820 TI - The druggable genome: an update. AB - Annotating the druggable genome estimates the potential maximum size of the playing field for current small-molecule drug design but It does not consider biologicals or future breakthroughs in medicinal chemistry or biology. PMID- 16376819 TI - Therapies for rheumatoid arthritis: hope springs eternal. PMID- 16376821 TI - More songs o' science. AB - A follow-up article concentrating on the use of songs on science in education and general entertainment. PMID- 16376822 TI - Keynote review: histamine H3 receptor antagonists reach out for the clinic. AB - Antagonists of the histamine H(1) and H(2) receptors have been successful as blockbuster drugs for treating allergic conditions and gastric ulcers, respectively. As such, histamine receptors have made a significant contribution to establishing G-Protein-coupled receptors as the favored drug targets of the industry. In this light, it can easily be understood that the discovery of a third histamine receptor subtype (H(3)R) in 1983 was greeted with considerable excitement. However, characterization of the H(3)R turned out to be far from trivial. In the past five years, molecular biology approaches have given fresh impetus to the H(3)R research field. As a result, H(3)R ligands are where they were anticipated to be 20 years ago: at the center of attention and on the verge of an anticipated breakthrough as the next generation of histaminergic blockbuster drugs. Here, we assess the status of the H(3)R medicinal chemistry programs of the various players in the field, as far as can be deduced from patent applications and scientific literature. PMID- 16376823 TI - Representativity of target families in the Protein Data Bank: impact for family directed structure-based drug discovery. AB - Analysis of the population of enzyme structures in the Protein Data Bank across all levels of the functional classification based on enzyme commission (EC) numbers reveals that, in spite of the almost exponential growth in the number of structures deposited, progress in achieving complete occupancy at all EC levels is relatively slow. Moreover, inspection of the distribution of the population among the members of the different enzyme families uncovers a strong bias towards enzymes widely recognized as therapeutically relevant targets. The low representativity levels identified in some target families warn on the current scope and applicability of structure-based approaches to family-directed strategies in drug discovery. PMID- 16376824 TI - Significance of release technology in tissue engineering. AB - Regenerative medical therapy has been expected to compensate for the therapeutic disadvantages of reconstructive surgery and organ transplantation, as well as offering a new therapeutic strategy. The objective of regenerative medical therapy is to induce the repair of defective tissues based on the natural healing potential of patients. For successful tissue regeneration, it is indispensable to provide cells with a local environment of artificial extracellular matrix where they can proliferate and differentiate efficiently. Tissue engineering is the key to this regeneration environment; release technology often enhances the in vivo stability of growth factors and related genes and prolongs the maintenance of biological functions for tissue regeneration. PMID- 16376825 TI - Novel and emerging therapies for asthma. AB - At present, there are a wide variety of novel and emerging therapeutic approaches for the treatment of asthma. Here, we will summarize these state-of-the-art approaches, including specific and nonspecific mediator inhibition-- a quest that has been on going for more than 25 years-- together with cytokine modulation in asthma (primarily attempting to modulate the Th2-Th1 balance in asthma), targeting cell recruitment, angiogenesis, signal transduction and gene transduction pathways. Finally, we will discuss the recently approved anti-IgE therapy for the treatment of allergic asthma and immune modulation using CpG oligodeoxynucleotides. PMID- 16376826 TI - From ligand design to therapeutic efficacy: the challenge for nicotinic receptor research. AB - S-Nicotine, the principal psychoactive constituent of Nicotiana tabacum, underpins addiction to tobacco smoking. Although tobacco consumption is a leading cause of death worldwide, nicotine itself is also proposed to have potential therapeutic benefits for a diverse range of conditions. Nicotine interacts with its cognate receptors in the central nervous system to exert a predominantly modulatory influence, making neuronal nicotinic receptors attractive therapeutic targets. Here, we focus on three natural products as lead compounds for drug discovery programs, nicotine, epibatidine and cytisine, and consider the aims and limitations that shape these drug discovery endeavors. PMID- 16376827 TI - Essential properties of drug-targeting delivery systems. AB - How, if at all, can drug delivery help to create ideal drugs? After four decades of trying, an effective site-specific drug-delivery system has not yet been developed. This review draws attention to the pharmacokinetic conditions that must be met to achieve a successful performance by site-selective drug-carrier delivery systems. In a drug-carrier approach, a drug is attached to a macromolecular carrier via a chemically labile linker. The carrier transports the drug to its site of action and releases it at the target site. For this simple approach to work, several fundamental conditions (nonspecific interactions, target site access, drug release and drug suitability) must be satisfied. The importance of these essential requirements, not always recognized in the development of drug-delivery systems, is discussed and illustrated by recent examples selected from the literature. PMID- 16376828 TI - Predicting protein druggability. AB - The ability to predict whether a particular protein can bind with high affinity and specificity to small, drug-like compounds based solely on its 3D structure has been a longstanding goal of structural biologists and computational scientists. The promise is that an accurate prediction of protein druggability can capitalize on the huge investments already made in structural genomics initiatives by identifying highly druggable proteins and using this information in target identification and validation campaigns. Here we discuss the potential utility of tools that characterize protein targets and describe strategies for the optimal integration of protein druggability data with bioinformatic approaches to target selection. PMID- 16376829 TI - Finding new drug targets in the 21st century. AB - The past 30 years have witnessed a steady decline in the number of new drug targets. This review concentrates on the initial process of target identification and argues that current problems have resulted from a decrease in clinical research, an overemphasis on the discovery of new targets through an understanding of the molecular causes of disease and the adoption of cell and animal models that are poor predictors of human disease. To resolve this situation, we argue for increased clinical research and show that an intervention at the physiological level, using drugs to target at the extracellular signalling pathways, will facilitate identification of novel drug targets in the 21st century. PMID- 16376830 TI - Storage and transmission of microarray images. AB - With the recent explosion of interest in microarray technology, massive amounts of microarray images are currently being produced. The storage and transmission of these types of data are becoming increasingly challenging. This article reviews the latest technologies that allow for the compression and storage of microarray images in dedicated database systems. PMID- 16376831 TI - Practical proteomic biomarker discovery: taking a step back to leap forward. AB - There is a pressing need for radically improved proteomic screening methods that allow for earlier diagnosis of disease, for systematic monitoring of physiological responses and for uncovering the fundamental mechanisms of drug action. Recent developments in proteomic technology offer tremendous, yet untapped, potential to yield novel biomarkers that are translatable to routine clinical use. Despite the significant conceptual promise of comparative proteomic profiling as a research platform for biomarker discovery, however, major hurdles remain for practical and clinical implementation. In particular, there is growing recognition that rigorous experimental design principles are urgently required to validate conclusively the unproven methodologies currently being touted. Debate and confusion persist about where the burden of proof lies: statistically, biologically or clinically? Moreover, there is no consensus about what constitutes a meaningful benchmark. An important question is how to achieve a scientifically rigorous, and therefore convincing, proof-of-concept that can be accepted by the field. Key analytical challenges related to these issues that must be addressed by the burgeoning biomarker community are discussed here. PMID- 16376836 TI - Pathogeny of archaic mycobacteria at the emergence of urban life in Egypt (3400 BC). AB - In a previous study, we extracted, amplified and sequenced a DNA fragment from bone lesions similar to those of bone tuberculosis in a predynastic skeleton (Egypt, around 3400 BC). This 65 kDa gene fragment encodes the surface of the mycobacterium responsible for the lesions. In the present study, we reconstructed the phylogenetic tree of the Mycobacterium family using this fragment and 51 sequences of pathogens and environmental mycobacteria. This reconstruction enabled us to polarise the phylogenic tree and to confirm the originality of the sequence and its ancestral character. According to a recent evolutionary scenario for the Mycobacterium tuberculosis complex, these data suggest the pathogenicity of an archaic mycobacterium at the emergence of urban life. This result could lead to a better understanding of present day evolutionary processes, especially the emergence or re-emergence of non-tuberculosis mycobacteria infection and disease. PMID- 16376837 TI - Genetic analysis of natural recombinant Brazilian Toxoplasma gondii strains by multilocus PCR-RFLP. AB - Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) at eight independent loci was used to determine the types (I, II or III) lineage of 20 Toxoplasma gondii strains isolated from humans and animals in Brazil. RH (type I, highly virulent), ME49 (type II, avirulent) and VEG (type III, avirulent) were used as reference strains. Differently from expected frequencies, all Brazilian strains showed to have recombinant genotypes, with typical alleles of types I, II or III at almost all loci assessed. The cB21-4 locus, a microsatellite marker, showed a higher allelic polymorphism with seven alleles among strains under analysis. Data have also shown that many Brazilian T. gondii strains presented a new haplotype at the L363 locus. When results of the eight loci were combined, 14 schizodemes were characterized out of the 20 T. gondii strains isolated in Brazil. The phenogram representing PCR-RFLP data separated Brazilian strains into two distinct genetic groups associated with murine virulence phenotype, termed groups I-A and I-B. Strains from group I-A (AS28, BV and N) that were highly virulent in BALB/c mice, were clustered with RH reference strain. Only those strains presented the haplotype I at the L363 locus, suggesting that this could be a possible marker of highly virulent strains. Strains from group I-B (cystogenic strains) showed a more heterogeneous behavior regarding virulence: a few of them (EGS, RAR, SAF, D5 and D6) were virulent, others (C4, P and D8) avirulent and most of them (D1, D2, D3, D4, D7, EFP, CH1, CH2 and CH3) intermediate virulent in mice. A significant linkage disequilibrium was observed in the population surveyed. However, the role of sexual recombination in the population structure of T. gondii in Brazil seems to be more central than in Europe and North America, where most studies have been performed. PMID- 16376838 TI - Identification and characterization of microsatellite markers in the Chagas disease vector Triatoma infestans (Heteroptera: Reduviidae). AB - Triatoma infestans, the main vector of Chagas disease in the southern cone countries, is the principal target of a regional elimination program. A better understanding of its dispersal, sources of reinfestation, and insecticide resistance is key to an effective control program. To address such problems, we identified and characterized 13 microsatellite loci of T. infestans. For each locus, primer sequences and PCR conditions are presented. Allele variability and frequency were analyzed in 59 T. infestans specimens from different rural communities in northwestern Argentina; nine loci were considered suitable for population genetic studies. Departure from Hardy-Weinberg equilibrium was detected in 10/13 loci with F(IS) values ranging from 0.04 to 0.91, indicating heterozygote deficit and a possible grade of sub-structure in the sample analyzed. Presence of null alleles in some loci cannot be discarded. The present work provides a promising tool to develop a population genetic study of natural populations of T. infestans in tandem with field studies and analyses of bug dispersal and the reinfestation process. PMID- 16376839 TI - A novel approach to characterise pathogen candidate genetic polymorphisms involved in clinical outcome. AB - Understanding the key factors influencing the clinical outcome of an infection is crucial for early diagnosis and optimised treatment. Despite widespread recognition of the importance of the genetics composition of pathogens, most efforts so far have focused on characterising disease and susceptibility genes in humans. Here, we propose a new flexible and powerful methodological framework to detect candidate genetic polymorphisms influencing clinical outcome from pathogen genomes. The rationale is to use well-supported clades in a phylogeny as statistical predictors for clinical outcomes rather than the individual polymorphisms themselves. This greatly increases the statistical power to detect candidate polymorphisms when analysing a large number of variable sites. In a second step, the candidate polymorphisms are recovered by characterising the polymorphisms that most strongly support the clades predicting the clinical outcome. The modelling approach further allows including host factors and testing for possible interactions between factors. We illustrate the approach by an application on a dataset of hepatitis B polymerase genes. The statistical model retains age at infection as well as six candidate polymorphisms as predictors for clinical outcome (acute, chronic and fulminant). The method is straightforward to apply and computationally effective. While the approach is focused on detecting candidate polymorphisms from pathogen genomes, the method might be more broadly applied for characterising the link between genotype and phenotype while statistically controlling for environmental factors. PMID- 16376840 TI - Origin and phylogeography of the Chagas disease main vector Triatoma infestans based on nuclear rDNA sequences and genome size. AB - For about half of all Chagas disease cases T. infestans has been the responsible vector. Contributing to its genetic knowledge will increase our understanding of the capacity of geographic expansion and domiciliation of triatomines. Populations of all infestans subcomplex species, T. infestans, T. delpontei, T. platensis and T. melanosoma and the so-called T. infestans "dark morph", from many South American countries were studied. A total of 10 and 7 different ITS-2 and ITS-1 haplotypes, respectively, were found. The total intraspecific ITS-2 nucleotide variability detected in T. infestans is the highest hitherto known in triatomines. ITS-1 minisatellites, detected for the first time in triatomines, proved to be homologous and thus become useful markers. Calculations show that ITS-1 evolves 1.12-2.60 times faster than ITS-2. Despite all species analyzed presenting the same n=22 chromosome number, a large variation of the haploid DNA content was found, including a strikingly high DNA content difference between Andean and non-Andean specimens of T. infestans (mean reduction of 30%, with a maximum of up to 40%) and a correlation between presence/absence of minisatellites and larger/smaller genome size. Population genetics analysis of the eight composite haplotypes of T. infestans and net differences corroborate that there are clear differences between western and eastern populations (60%), and little genetic variation among populations (1.3%) and within populations (40%) within these two groups with migration rates larger than one individual per generation corresponding only to pairs of populations one from each of these groups. These values are indicative either of a large enough gene flow to prevent population differentiation by drift within each geographic area or a very recent spread, the latter hypothesis fitting available data better. Phylogenetic trees support a common ancestor for T. infestans and T. platensis, an origin of T. infestans in Bolivian highlands and two different dispersal lines, one throughout Andean regions of Bolivia and Peru and another in non-Andean lowlands of Chile, Paraguay, Argentina, Uruguay and Brazil. PMID- 16376842 TI - Proteomic studies of Plasmodium knowlesi SICA variant antigens demonstrate their relationship with P. falciparum EMP1. AB - Malaria variant antigens are encoded by large multigene families and expressed on the surface of infected erythrocytes. The Plasmodium knowlesi Schizont-infected cell agglutination (SICA) antigens are encoded by the SICAvar multigene family, and the P. falciparum erythrocyte membrane protein-1 (PfEMP1) antigens are encoded by the var gene family. Although these variant antigens share many fundamental features, P. knowlesi and P. falciparum are phylogenetically distantly related, and so far a significant level of sequence identity has not been observed in alignments of either the SICAvar and var gene families or their encoded proteins. In support of their orthologous relationship, however, here we demonstrate through proteomic analysis that the P. knowlesi SICA variant antigens share significant common sequences with P. falciparum EMP1 molecules. As many as forty P. knowlesi SICA peptides show identity with a particular P. falciparum EMP1, mapping throughout all characterized domains, including the externally exposed cysteine-rich domains that are characteristic of both proteins. These findings provide further validation of the classical in vivo P. knowlesi-rhesus monkey model system for advancing our understanding of the immunobiology of antigenic variation and variant antigen gene expression in Plasmodium. PMID- 16376843 TI - The impact of a novel educational curriculum for first-time DUI offenders on intermediate outcomes relevant to DUI recidivism. AB - The Preventing Alcohol-Related Convictions (PARC) program is a novel educational curriculum for first-time DUI offenders, with the ultimate goal of reducing DUI recidivism. It differs from traditional DUI education and prevention programs in that it does not suggest to DUI offenders that they must abstain from alcohol entirely or control their drinking to prevent a future DUI; rather, it teaches students to prevent a future DUI by not driving their cars to drinking events. Thus, the emphasis of the curriculum is on controlling driving rather than controlling drinking to avoid future DUI convictions. The implementation of the program is ongoing throughout the state of Florida. The current randomized study focused on intermediate outcomes relevant for DUI recidivism; specifically, individuals' readiness for change regarding drinking and driving, and their endorsement of a PARC planning and action approach (controlling driving) versus a traditional approach (controlling drinking). The current research demonstrated that the PARC program is effective in moving participants toward more readiness for change and toward a strategy of planning ahead to avoid driving to any venue in which drinking may occur. Future research will assess the ultimate effect on DUI recidivism. PMID- 16376841 TI - Patterns of Eurasian HSV-1 molecular diversity and inferences of human migrations. AB - Following our recent report of high levels of recombination and geographic structuring amongst isolates from two populations, we have investigated global patterns of herpes simplex virus type 1 (HSV-1) molecular diversity using population samples from six countries in Europe, Asia and Africa. Sequence comparisons show that HSV-1 from Kenya is both highly diverse and distinct from either European or Asian HSV-1. HSV-1 populations are much more highly differentiated than human populations at the same geographic scales, with 35% of total variation at the level of inter-population comparisons, a difference likely to be due to higher rates of both mutation and genetic drift in HSV-1 than in equivalent human data. There is substantial differentiation between northwestern European HSV-1 populations and those from East Asia, and while patterns of British and Swedish HSV-1 variation were indistinguishable, differentiation was detectable amongst Chinese, Korean and Japanese HSV-1 samples, in spite of their lower overall diversity. The program Structure was used to reconstruct ancestral Eurasian lineages, which we estimated to have originated approximately 60,000 years ago. A specific pattern detected amongst East Asian HSV-1 isolates is currently best explained by the two waves of migration responsible for the peopling of Japan. PMID- 16376844 TI - The potentiation of two components of the reach-to-grasp action during object categorisation in visual memory. AB - Stimulus-Response Compatibility Effects have been reported for several components of the reach-to-grasp action during visual object recognition [Tucker, M., & Ellis, R. (1998). On the relations between seen objects and components of potential actions. Journal of Experimental Psychology: Human Perception and Performance, 24, 830-846; Ellis, R., & Tucker, M. (2000). Micro-affordance: The potentiation of actions by seen objects. British Journal of Psychology, 91, 451 471; Tucker, M., & Ellis, R. (2001). The potentiation of grasp types during visual object categorization. Visual Cognition, 8, 769-800; Creem, S. H., & Proffitt, D. R. (2001). Grasping objects by their handles: A necessary interaction between cognition and action. Journal of Experimental Psychology: Human Perception and Performance, 27, 218-228; Craighero, L. Bello, A. Fadiga, L., & Rizzolatti, G. (2002). Hand action preparation influences the responses to hand pictures. Neuropsychologia, 40, 492-502]. The present study investigates compatibility effects for two elements of reach-to-grasp action during the visual mental imagery of objects-the compatibility of an object for grasping with a power and precision grasp, and the orientation of an object (left/right) for grasping by a particular hand (left/right). Experiment 1 provides further evidence for compatibility effects of a 'seen' object for grasping with a power and precision grasp. The experiment shows that compatibility effects are obtainable when an object is presented in an array of four objects and not just on its own. Experiment 2 provides evidence that compatibility effects of an object for grasping with a power and precision grasp can also be observed when participants make an action response to an object 700 ms after it has been removed from view. Experiment 3 investigates compatibility effects for the orientation of an object for grasping by a particular hand during visual mental imagery, but finds no evidence for such effects. The findings are discussed in relation to two arguments put forward to reconcile ecological and representational theories of visual object recognition. PMID- 16376845 TI - [Surgical treatment of the single non traumatic perforation of small bowel: excision-suture or resection-anastomosis]. AB - OBJECTIVE: To evaluate morbidity and mortality following excision-suture and resection-anastomosis for single non traumatic perforations of small bowel (SNTPB). METHODS: From July 2002 to June 2003, a simple blind randomized study comparing excision-suture with resection-anastomosis SNTPB. RESULTS: Of the 125 patients included, 112 were operated by surgeons on training (89.6%). The perforation sat on the antimesenteric edge of the last ileal portion with an average diameter of 0.8 cm (extreme 0.1 and 4 cm). An excision-suture was performed 66 times (52%) including 5 times by an experienced surgeon. 56 patients had simple continuations (45.2%). Morbidity concerned 68 patients (54,8%). 06 patients died of general complications (4.8%). The technique did not influence the mortality. All dead patients were operated by surgeons on training, P=0.25. The complications were significantly more frequent after resection-anastomosis (79.7%) than after excision-suture (32.3%). The difference was significant for the parietal abscesses (P=0,01), the exteriorized fistulas (P=0.04), the septic shocks (P=0.05). CONCLUSION: Whereas mortality was not influenced by the technique, the postoperative course was more complicated after resection anastomosis (performed in majority by less experienced surgeons). We recommend excision-suture to repair SNTPB. PMID- 16376846 TI - [Endoscopic treatment of Barrett's oesophagus]. AB - Aims of the treatment of Barrett's oesophagus (BO) are disappearance of symptoms and inflammatory complications of gastro-oesophageal reflux disease (GORD), prevention of occurrence of dysplasia and adenocarcinoma, and early treatment of high-grade dysplasia (HGD) and adenocarcinoma. Anti-secretory treatment with proton-pump inhibitors (PPI) must result in disappearance of both symptoms and oesophagitis. The only correction of symptoms, as well as normalization of pHmetry, are not considered as adequate criteria for efficiency of treatment. It has not been demonstrated that treatment with PPI prevented occurrence of dysplasia and adenocarcinoma so the only BO is not an indication for treatment with PPI, which results in only partial regression of height and/or surface of BO. Endoscopic ablation of BO, combined with PPI, allows complete regression of intestinal metaplasia in about 50% of cases. Photodynamic therapy (PDT) seems the best technique for treatment of HGD and mucosal adenocarcinoma. This treatment is not indicated in case of low-grade dysplasia, since its benefit on survival is less clear than for HGD. Endoscopic treatment does not suppress the need for prolonged endoscopic follow-up since BO recurs in approximately one third of patients. For HGD isolated or associated with mucosal adenocarcinoma (proven by endoscopic ultrasound), endoscopic treatment can consist in mucosectomy or ablation with PDT or plasma argon coagulation (PAC) of these lesions if localized, possibly followed by complete ablation of BO by PAC, and always associated with an efficient treatment of GORD by PPI or anti-reflux surgery. Submucosal adenocarcinomas must be treated by oesophagectomy if allowed by the general condition of the patient. PMID- 16376847 TI - [The Association Francaise de Chirurgie (AFC) colorectal index: a reliable preoperative prognostic index in colorectal surgery]. AB - INTRODUCTION: Evaluation of outcome after colorectal surgery is always necessary. A new index which permits to appreciate preoperatively postoperative mortality after colorectal resection in colorectal cancer (CRC) and in diverticular disease has been published (i.e., Association Francaise de Chirurgie, AFC colorectal index). PATIENTS AND METHODS: From November 2002 to July 2004, in-hospital mortality was analysed on 253 patients who underwent colic resection (N = 220, 87%) or rectal resection, with anastomosis (N = 175, 70%). Mortality was analysed according to emergency resection, neurological co morbidity, lost of weight more than 10% of weight, age older than 70 years. RESULTS: Mean age of patients was 63 +/- 18 years (17-92) (45% older than 70 years), 26% of patients were ASA >or= III, 35% underwent surgery in emergency, and 12% underwent laparoscopic surgery. One hundred and fifteen (45%) patients underwent surgery for CRC and 50 (20%), for diverticular disease and 11 patients underwent surgery for ischemic colitis. Overall mortality rate was 10% (N = 26), it was 19% in emergency surgery versus 5% after elective surgery. Global morbidity was 38%, percentage of anastomotic leak was 8% (N = 14/175), reoperation was necessary in 14%. The mean length of stay was 13 +/- 8 days. Ten percent of patients necessitated unplanned readmission. After surgery for CCR or diverticular disease. -i) overall mortality was 9% - ii) among patients who had 0, 1, 2, or 3 predictive risk factors of mortality; mortality was 0% , 5% 15% and 33%. After surgery for other aetiology than CCR or diverticular disease, among patients who had 0, 1, 2, or 3 predictive risk factors of mortality; mortality was 0%, 12% 36% and 25%. CONCLUSIONS: These results showed the reproducibility of the AFC colorectal index and its potential application in all aetiologies after colorectal surgery. PMID- 16376848 TI - [Management of post-traumatic isolated adrenal haematoma]. AB - We report three cases of isolated adrenal haematoma diagnosed at admission to the Emergency Department. Post-traumatic adrenal haematoma have been observed in 2% of patients with abdominal injury and can be life threatening in cases of bilateral haemorrhage with risk of acute adrenocortical insufficiency. Diagnosis remains difficult due to absence of clinical and biological specific parameters. Therefore, abdominal CT examination should be the gold standard in management of abdominal trauma patients. PMID- 16376849 TI - [Barrett's oesophagus: endoscopic diagnosis and follow-up]. AB - Barrett's oesophagus (BO), or replacement of the squamous mucosa by a specialized intestinal metaplasia due to gastro-oesophageal reflux disease (GORD), predisposes to adenocarcinoma. It is estimated that 6 to 12% of patients undergoing GI endoscopy have short BO (< 3 cm), and 1% have a long BO. Macroscopic diagnosis of BO is sometimes difficult and, in case of doubt, endoscopy should be redone after a period of efficient anti-secretory treatment. Diagnosis of BO is histological and should be confirmed by biopsies. The incidence of adenocarcinoma is globally estimated at 0.5% patient by year of follow-up, and exists for both short and long BO. Due to this low incidence, screening for BO is only justified in patients at high risk for adenocarcinoma (male gender, age > 50 ans, old GORD in a young patient). Low-grade dysplasia (LGD) then high-grade dysplasia (HGD) precedes adenocarcinoma. Histological diagnosis of LGD is difficult: the main cause of confusion is inflammation so diagnosis of LGD must be confirmed after a 3-month high-dose anti-secretory treatment. Diagnosis of HGD is easier but multiple biopsies are needed to determine the focal or multifocal disposition of HGD. The benefit of follow-up of BO is debated. Aged patients should be followed only if dysplasia is present. When dysplasia is absent, an endoscopic control with biopsies is desirable within 3 to 5 years. In case of dysplasia, the latter must be confirmed by another examination of biopsies, particularly in case of suspicion of HGD and after antisecretory treatment. In case of LGD, endoscopy with biopsies should be redone 6 months later to screen for HGD, then every year if LGD is confirmed. In case of HGD, the 5-year risk of cancer is 60% so surgical or endoscopic treatment is usually proposed. If HGD follow-up is decided, it should be performed on a 3- to 6-month basis. PMID- 16376851 TI - Eating patterns and breakfast consumption in obese patients with binge eating disorder. AB - This study examined eating patterns and breakfast consumption, and their relationships to weight and binge eating, in obese individuals with binge eating disorder (BED). One-hundred seventy-three consecutively evaluated men (n=46) and women (n=127) with BED were administered semi-structured interviews and self report measures to assess the frequency of meals and snacks eaten, as well as binge eating and eating disorder features. Overall, those who consumed more frequent meals, particularly breakfast, and snacks, weighed less. Breakfast, which was eaten on a daily basis by less than half of participants (n=74; 43%), was the least frequently eaten meal of the day. Participants (n=56; 32%) who ate three meals per day weighed significantly less, and had significantly fewer binges, than participants (n=117; 68%) who did not regularly eat three meals per day. Thus, eating more frequently, having breakfast and consuming three meals every day, have potentially important clinical applications for the treatment of BED given that the effectiveness of specific interventions within treatments for BED are unknown, and that weight loss outcome for BED has been poor. PMID- 16376850 TI - Upstream stimulatory factor 1 transactivates the human gene promoter of the cardiac isoform of acetyl-CoA carboxylase. AB - E-box cis-elements act as binding sites for upstream stimulatory factors (USFs), putative glucose-responsive transcriptional modulators. Since four E-boxes were identified on the human ACCbeta promoter, we hypothesized that USF1 induces ACCbeta expression in a glucose-dependent manner. Here, murine cardiac ACCbeta expression was significantly increased in response to high carbohydrate re feeding after fasting. However, transfection studies showed no difference in ACCbeta promoter activity in neonatal cardiomyocytes and CV-1 fibroblasts after low (5.5mM) and high (25 mM) glucose exposure. USF1 overexpression significantly increased ACCbeta promoter activity in both cell lines under low glucose conditions. With high glucose exposure, USF1 further induced ACCbeta promoter activity only in CV-1 fibroblasts. USF1-induced ACCbeta promoter responsiveness was markedly attenuated when co-transfecting cardiomyocytes with a -93/+65 or 38/+65 promoter deletion construct (lacking E-boxes 1-3). Thus, USF1 transactivates the human ACCbeta promoter in the heart, likely through an E-box cis-element located close to the transcription start site. PMID- 16376852 TI - C/EBPalpha-dependent induction of glutathione S-transferase zeta/maleylacetoacetate isomerase (GSTzeta/MAAI) expression during the differentiation of mouse fibroblasts into adipocytes. AB - Western blot analysis of 3T3-L1 adipocyte proteins using an anti-C/EBPalpha antibody detected a 24kD polypeptide in addition to the expected 42 and 30kD isoforms of C/EBPalpha. Mass spectrometric sequencing of the protein following its purification by HPLC and preparative 2D gel electrophoresis identified it as glutathione S-transferase zeta/maleylacetoacetate isomerase (GSTzeta/MAAI). Expression of GSTzeta/MAAI mRNA and protein was induced during the terminal phase of adipogenesis in 3T3-L1 preadipocytes. Ectopic expression of PPARgamma2 in NIH 3T3 fibroblasts exposed to insulin and troglitazone-induced perilipin production, but was incapable of activating GSTzeta/MAAI unless C/EBPalpha was also expressed. Similarly, ectopic expression of C/EBPalpha in PPARgamma +/- or PPARgamma -/- MEFs demonstrated that the C/EBPalpha-dependent induction of GSTzeta/MAAI production was dependent on expression of endogenous PPARgamma. These data suggest a role for GSTzeta/MAAI in mature adipocytes that may be responsive to the thiazolidinedione class of insulin sensitizing PPARgamma ligands. PMID- 16376853 TI - Detection of presenilin-1 homodimer formation in intact cells using fluorescent lifetime imaging microscopy. AB - Presenilin-1 (PS1) is a multipass transmembrane domain protein, which is believed to be the catalytic component of the gamma-secretase complex. The complex is comprised of four major components: PS1, nicastrin, Aph-1, and Pen-2. The exact stoichiometric relationship between the four components remains unclear. It has been shown that gamma-secretase exists as high molecular weight complexes, suggesting the possibility of dimer/multimer formation. We combined a biochemical approach with a novel morphological microscopy assay to analyze PS1 dimer formation and subcellular distribution in situ, in intact mammalian cells. Both coimmunoprecipitation and fluorescent lifetime imaging microscopy approaches showed that wildtype PS1 molecules form dimers. Moreover, PS1 holoproteins containing the D257A mutation also come into close enough proximity to form a dimer, suggesting that cleavage within the loop is not necessary for dimer formation. Taken together these data suggest that PS1 dimerization occurs during normal PS1 function. PMID- 16376854 TI - Lipid membrane interaction and antimicrobial activity of GsMTx-4, an inhibitor of mechanosensitive channel. AB - GsMTx-4, a polypeptide from the spider Grammostola spatulata, is an inhibitor of mechanosensitive channels. It is known to interact with lipid membranes, suggesting it partitions into the membrane to alter the channel gating, but the effect of the membrane charge on GsMTx-4 activity remains unknown. In this study, we found that GsMTx-4 more effectively interacts with anionic lipids than zwitterionic ones. The effect of GsMTx-4 on negatively charged membranes was similar to that of the antimicrobial peptide melittin, which led us to assess GsMTx-4's antimicrobial activity. Interestingly, we found that, in contrast to other neurotoxins, GsMTx-4 exhibited antimicrobial properties and was more active against Gram-positive than Gram-negative bacteria. These results suggest that GsMTx-4 exerts its antimicrobial effect by altering the packing of the membrane and/or inhibiting mechanosensitive channels. These findings could point the way towards a new class of antimicrobial peptides. PMID- 16376855 TI - Isolation and the characterization of the degradation products of the mediator ABTS-derived radicals formed upon reaction with polyphenols. AB - Two degradation products were obtained from the incubation of the widely used 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), ABTS, radical cations with the polyphenols, (+)-catechin, (-)-epicatechin, and phloroglucinol in acetate buffer (pH 5). The products were purified by reversed-phase chromatography and characterized by UV-visible detection, mass spectrometry, and (1)H NMR spectroscopy. The data allowed us to identify the degradation products as 3-ethyl-6-sulfonate benzothiazolinone imine and the corresponding sulfoxide, 3 ethyl-6-sulfonate benzothiazolone. Elemental composition strongly supported the proposed structures. Our results unequivocally demonstrated that ABTS radicals are not as stable as usually claimed because they could be degraded upon interaction with polyphenols, in addition to being reduced by these antioxidants back to the parent compound. Therefore, it is concluded that caution must be exercised in using ABTS radicals as a basis for the evaluation of antioxidant capacities of pure compounds and/or complex mixtures. PMID- 16376856 TI - Granulocyte colony-stimulating factor treatment enhances the efficacy of cellular cardiomyoplasty with transplantation of embryonic stem cell-derived cardiomyocytes in infarcted myocardium. AB - We tested the hypothesis that granulocyte colony-stimulating factor (G-CSF) administration would enhance the efficacy of cellular cardiomyoplasty with embryonic stem (ES) cell-derived cardiomyocytes in infarcted myocardium. Three weeks after myocardial infarction by cryoinjury, Sprague-Dawley rats were randomized to receive either an injection of medium, ES cell-derived cardiomyocyte transplantation, G-CSF administration, or a combination of G-CSF administration and ES cell-derived cardiomyocyte transplantation. Eight weeks after treatment, the cardiac tissue formation, neovascularization, and apoptotic activity in the infarct regions were evaluated by histology and immunohistochemistry. The left ventricular (LV) dimensions and function of the treated heart were evaluated by echocardiography. Transplanted ES cell-derived cardiomyocytes survived and participated in the myocardial regeneration in the infarcted heart. A combination of G-CSF treatment and ES cell-derived cardiomyocyte transplantation significantly promoted angiogenesis and reduced the infarct area and cell apoptosis in the infarcted myocardium compared with ES cell derived cardiomyocyte transplantation alone. The combination therapy also attenuated LV dilation, as compared with ES cell-derived cardiomyocyte transplantation alone. G-CSF treatment can enhance the efficacy of cellular cardiomyoplasty by ES cell-derived cardiomyocyte transplantation to treat myocardial infarction. PMID- 16376857 TI - Clonal proliferation of multipotent stem/progenitor cells in the neonatal and adult salivary glands. AB - Salivary gland stem/progenitor cells are thought to be present in intercalated ductal cells, but the fact is unclear. In this study, we sought to clarify if stem/progenitor cells are present in submandibular glands using colony assay, which is one of the stem cell assay methods. Using a low-density culture of submandibular gland cells of neonatal rats, we developed a novel culture system that promotes single cell colony formation. Average doubling time for the colony forming cells was 24.7 (SD=+/-7.02)h, indicating high proliferative potency. When epidermal growth factor (EGF) and hepatocyte growth factor (HGF) were added to the medium, the number of clonal colonies increased greater than those cultured without growth factors (13.2+/-4.18 vs. 4.5+/-1.73). The RT-PCR and immunostaining demonstrated expressing acinar, ductal, and myoepithelial cell lineage markers. This study demonstrated the presence of the salivary gland stem/progenitor cells that are highly proliferative and multipotent in salivary glands. PMID- 16376858 TI - Characterization of enzymatic properties of human ribonucleotide reductase holoenzyme reconstituted in vitro from hRRM1, hRRM2, and p53R2 subunits. AB - Ribonucleotide reductase (RR) is a highly regulated enzyme in the deoxyribonucleotide synthesis pathway. RR is responsible for the de novo conversion of ribonucleoside diphosphates to deoxyribonucleoside diphosphates, which are essential for DNA synthesis and repair. Besides two subunits, hRRM1 and hRRM2, p53R2 is a newly identified member of RR family that is induced by ultraviolet light in a p53-dependent manner. To understand the molecular interaction of RR subunits, we employed a eukaryotic expression system to express and purify all three subunits. After in vitro reconstitution, the results of [(3)H]CDP reduction assay showed that both eukaryotic recombinant hRRM2 and p53R2 proteins could interact with hRRM1 to form functional RR holoenzyme. The reconstituted RR activity was time-dependent and the reaction rate reached the plateau phase after 40min incubation. No matter the concentration, RR holoenzyme reconstituted from p53R2 and hRRM1 could only achieve about 40-75% kinetic activity of that from hRRM2 and hRRM1. The synthetic C-terminal heptapeptide competition assays confirmed that hRRM2 and p53R2 share the same binding site on hRRM1, but the binding site on hRRM1 demonstrated higher affinity for hRRM2 than for p53R2. In allosteric regulation assay, the effect of activation or inhibition of hRRM1 with ATP or dATP suggested that these effectors could regulate RR activity independent of different RR small subunits. Taken together, the eukaryotic expression system RR holoenzyme will provide a very useful tool to understand the molecular mechanisms of RR activity and the interactions of its subunits. PMID- 16376859 TI - Gene expression profiles of the rat brain both immediately and 3 months following acute sarin exposure. AB - We have studied sarin-induced global gene expression patterns at an early time point (15 min; 0.5xLD50) and a later time point (3 months; 1xLD50) using Affymetrix: Rat Neurobiology U34 chips in male, Sprague-Dawley rats and have identified a total of 65 (early) and 38 (late) genes showing statistically significant alterations from control levels at 15 min and 3 months, respectively. At the early time point, those that are classified as ion channel, cytoskeletal and cell adhesion molecules, in addition to neuropeptides and their receptors predominated over all other groups. The other groups included: cholinergic signaling, calcium channel and binding proteins, transporters, chemokines, GABAnergic, glutamatergic, aspartate, catecholaminergic, nitric oxide synthase, purinergic, and serotonergic signaling molecules. At the late time point, genes that are classified as calcium channel and binding proteins, cytoskeletal and cell adhesion molecules and GABAnergic signaling molecules were most prominent. Seven molecules (Ania-9, Arrb-1, CX-3C, Gabab-1d, Nos-2a, Nrxn-1b, PDE2) were identified that showed altered persistent expression in both time points. Selected genes from each of these time points were further validated using semi quantitative RT-PCR approaches. Some of the genes that were identified in the present study have been shown to be involved in organophosphate-induced neurotoxicity by both other groups as well as ours. Principal component analysis (PCA) of the expression data from both time points was used for comparative analysis of the gene expression, which indicated that the changes in gene expression were a function of dose and time of euthanasia after the treatment. Our model also predicts that besides dose and duration of post-treatment period, age and possibly other factors may be playing important roles in the regulation of pathways, leading to the neurotoxicity. PMID- 16376860 TI - Physical activity and restlessness correlate with leptin levels in patients with adolescent anorexia nervosa. AB - BACKGROUND: In food-restricted rats, leptin suppresses semistarvation-induced hyperactivity (SIH) and decreases exploratory behavior. Leptin ameliorates anxiety-related movement in ob/ob mice. In this study, we assessed the relationship between leptin and qualities of physical activity and restlessness in acute anorexia nervosa (AN). METHODS: Serum leptin, body mass index (BMI), % body fat, and self- and expert-ratings of qualities of physical activity and restlessness were assessed in 26 inpatients with acute AN. Accelerometry was also performed. Regression analyses were used to predict activity and restlessness using BMI, % body fat, and leptin levels as predictor variables. RESULTS: Leptin levels significantly contributed to the prediction of all measures of activity and restlessness. CONCLUSIONS: This is the first study linking hypoleptinemia in AN patients to subjective and objective measures of higher physical activity and motor and inner restlessness. Leptin may directly or indirectly (or both) influence behaviors and cognitions contributing to hyperactivity and motor restlessness. PMID- 16376861 TI - Short- and long-term changes in anterior cingulate activation during resolution of task-set competition. AB - Alternating between task sets involves detection that the current task set is unfavorable, initiation of a change in set, and application of the new task set while fine-tuning to optimally adjust to the demands of the environment. Functional magnetic resonance imaging (fMRI) studies of cognitive flexibility consistently report activation of the anterior cingulate cortex and/or adjacent pre-supplementary motor regions (ACC/pre-SMA, medial Brodmann's areas 24/32/6), suggesting that these cortical regions are involved in switching task set. In the current study, our objective was to probe whether ACC/pre-SMA activation would decrease for a number of trials following a switch in task set, implying longer term involvement in fine-tuning adjustments. By measuring activation when switching between word reading and color naming in response to Stroop stimuli, ACC/pre-SMA activation was observed when actively countering the influence of the irrelevant task set, and this activation decreased as a function of the number of trials since a task switch. Basal ganglia and thalamic regions also displayed a decreased response over successive trials after task switches. These findings suggest that the ACC/pre-SMA are not only involved in generating a new course of action, but are also involved (along with subcortical regions) in fine-tuning operations that resolve competition between task sets over subsequent repetitions of the same task. PMID- 16376862 TI - Investigation of changes in global gene expression in the frontal cortex of early weaned and socially isolated piglets using microarray and quantitative real-time RT-PCR. AB - We hypothesize that early-weaned piglets experience aberrant expression of stress responsive genes in the frontal cortex, a key brain area involved in cognitive function and behavior organization. To test this hypothesis, female early-weaned piglets (EW; n = 6) were weaned 10 days after birth, while non-weaned piglets (NW; n = 6) were left with their dams. Half of EW (n = 3) and NW (n = 3) animals were socially isolated (SI) for 15 min at 12 days of age, when all animals (n = 12) were euthanized and tissue collected. The effects of EW and SI were examined by gene expression profiling using cDNA microarray hybridizations, generated from a porcine brain cDNA library. A total of 103 genes were differentially expressed (P < 0.05, fold change >1.25) among four direct comparisons. Forty-two genes had known functions, from which 24 showed relevant brain-related functions. Quantitative real-time polymerase chain reaction (Q-RT-PCR) was used to confirm regulation of expression of a subset of 6 genes with important brain functions, selected from the microarray outcomes. In non-weaned animals, a significant suppression of mRNA abundance for carboxypeptidase E, 14-3-3 protein and phosphoprotein enriched in astrocytes 15 kDa was observed in response to SI. Also, in early-weaned animals, diazepam binding inhibitor and actin-related protein 2/3 complex mRNA levels were suppressed in response to SI. Results suggest that social isolation of non- and early-weaned piglets may impact expression of genes involved in regulation of neuronal function, development, and protection in the frontal cortex of young pigs. PMID- 16376863 TI - Stress-induced heat shock protein 27 expression and its role in dorsal root ganglion neuronal survival. AB - Heat shock protein 27 (Hsp27), a molecular chaperone ubiquitously expressed in many cell types, has been shown to play a role in protecting neurons from cellular stresses. Unlike adult DRG neurons in vitro, neonatal DRG neurons require NGF for survival; withdrawal of NGF results in apoptosis of a majority of neonatal neurons. We hypothesized that Hsp27 contributes to the neurotrophin independent survival of adult DRG neurons. Constitutive Hsp27 expression is higher in adult DRG neurons compared to neonates, although both upregulate Hsp27 expression after heat shock (HS). We found that increasing endogenous Hsp27 by HS in neonatal neurons was able to inhibit NGF withdrawal-induced apoptosis. Heat shock of adult and neonatal neurons also resulted in Akt activation, which could be a mechanism for the increased survival. Hsp27 siRNA treatment of adult neurons effected a decreased expression of Hsp27, which correlated with increased apoptosis in these neurons. Downregulation of Hsp27 via siRNA also blocked the HS induced rescue of neonatal neurons after NGF withdrawal. These results indicate that physiologically induced upregulation of Hsp27 is sufficient to provide some degree of neuronal protection. Further, this induction appears to be regulated by the transcriptional activation of HSF1 as shown by HSF1 nuclear translocation and by EMSA analyses of HSF1 binding to nuclear protein. PMID- 16376864 TI - Identification of forkhead transcription factors in cortical and dopaminergic areas of the adult murine brain. AB - The murine forkhead family of transcription factors consists of over 30 members, the vast majority of which is important in embryonic development. Implicated in processes such as proliferation, differentiation and survival, forkhead factors show highly restricted expression patterns. In search for forkhead genes expressed in specific neural systems, we identified multiple family members. We performed a detailed expression analysis for Foxj2, Foxk1 and the murine orthologue of the human ILF1 gene, which show a remarkable preference for complex cortical structures. In addition, a comprehensive examination of forkhead gene expression in dopamine neurons of the ventral tegmental area and substantia nigra pars compacta, revealed Ilf1 as a novel transcriptional regulator in midbrain dopamine neurons. These forkhead transcription factors may play a role in maintenance and survival of developing and adult neurons. PMID- 16376865 TI - Microarray profiling of gene expression patterns in adult male rat brain following acute progesterone treatment. AB - Progesterone can influence various behaviors in adult male rats, however, little is known about which particular genes are regulated by progesterone in the male rat brain. Using focused microarray technology, we where able to define a subset of genes that are responsive to progesterone. Nylon membrane-based cDNA microarrays were used to profile gene expression patterns in the preoptic area/mediobasal hypothalamus (POA/MBH) of male rat brain 7 h following a single injection of progesterone. RNA was isolated from the brains of 6 male rats injected with progesterone and 6 male rats injected with sesame oil. Next, we hybridized the RNA from each animal to individual cDNA microarrays that contained more than 100 target genes, all of which are involved in cAMP and or calcium signaling pathways. Direct side-by-side comparison of all 12 arrays revealed differences in the expression patterns of 12 different genes. We confirmed the data gathered from the arrays on 4 different genes using Real-Time PCR. These data begin to outline the important role played by progesterone in mediating changes in gene expression within the male brain. PMID- 16376867 TI - New regioselective derivatives of sucrose with amino acid and acrylic groups. AB - We report here a range of new sucrose derivatives obtained from '3-ketosucrose' in aqueous medium with few reaction steps. As an intermediate, 3-amino-3-deoxy alpha-D-allopyranosyl beta-D-fructofuranoside (1) was obtained via the classical route of reductive amination with much improved yield and high stereoselectivity. Building blocks for polymerization were synthesized by introduction of acrylic type side chains, for example, with methacrylic anhydride. Corresponding polymers were synthesized. Aminoacyl and peptide conjugates were obtained through conventional peptide synthesis with activated and protected amino acids. Deprotection yielded new glycoderivatives having an unconventional substitution pattern, namely 3-(aminoacylamino) allosaccharides. Both mono- and di-peptide conjugates of allosucrose have been synthesized. PMID- 16376868 TI - Chitosan bicomponent nanofibers and nanoporous fibers. AB - Nanofibers with average diameters between 20 and 100nm have been prepared by electrospinning of 82.5% deacetylated chitosan (Mv=1600 kDa) mixed with poly(vinyl alcohol) (PVA, Mw=124-186 kDa) in 2% (v/v) aqueous acetic acid. The formation of bicomponent fibers was feasible with 3% concentration of solution containing up to an equal mass of chitosan. Finer fibers, fewer beaded structures and more efficient fiber formation were observed with increasing PVA contents. Nanoporous fibers could be generated by removing the PVA component in the 17/83 chitosan/PVA bicomponent fibers with 1M NaOH (12 h). Fiber formation efficiency and composition uniformity improved significantly when the molecular weight of chitosan was halved by alkaline hydrolysis (50 wt% aqueous NaOH, 95 degrees C, 48 h). The improved uniform distribution of chitosan and PVA in the bicomponent fibers was attributed to better mixing mostly due to the reduced molecular weight and to the increased deacetylation of the chitosan. PMID- 16376869 TI - Nitric oxide is a preconditioning mimetic and cardioprotectant and is the basis of many available infarct-sparing strategies. AB - Ischemic preconditioning is a powerful infarct-sparing intervention. Intensive investigations have revealed many of the signaling steps used to elicit this protection. One of the steps involves activation of nitric oxide synthase (NOS) by phosphorylation, with the production of NO and subsequent activation of guanylyl cyclase, production of cGMP, activation of protein kinase G, opening of mitochondrial KATP channels, and generation of reactive oxygen species. The latter act as second messengers to activate critical kinase cascades that trigger entrance into the preconditioned state. Thus, NO exposure before ischemia can act as a powerful preconditioning mimetic. Elevating NO just prior to or at reperfusion can still be an effective cardioprotective strategy. Activation of NOS or production of NO can be done pharmacologically with exogenous agents to trigger this cascade. Many of these strategies are already available and safe. PMID- 16376870 TI - Functional effects of protein kinase C-mediated myofilament phosphorylation in human myocardium. AB - OBJECTIVE: In human heart failure beta-adrenergic-mediated protein kinase A (PKA) activity is down-regulated, while protein kinase C (PKC) activity is up regulated. PKC-mediated myofilament protein phosphorylation might be detrimental for contractile function in cardiomyopathy. This study was designed to reveal the effects of PKC on myofilament function in human myocardium under basal conditions and upon modulation of protein phosphorylation by PKA and phosphatases. METHODS: Isometric force was measured at different [Ca(2+)] in single permeabilized cardiomyocytes from non-failing and failing human left ventricular tissue. Basal phosphorylation of myofilament proteins and the influence of PKC, PKA, and phosphatase treatments were analyzed by one- and two-dimensional gel electrophoresis, Western immunoblotting, and ELISA. RESULTS: Troponin I (TnI) phosphorylation at the PKA sites was decreased in failing compared to non-failing hearts and correlated well with myofilament Ca(2+) sensitivity (pCa(50)). Incubation with the catalytic domain of PKC slightly decreased maximal force under basal conditions, but not following PKA and phosphatase pretreatments. PKC reduced Ca(2+) sensitivity to a larger extent in failing (DeltapCa(50)=0.19+/ 0.03) than in non-failing (DeltapCa(50)=0.08+/-0.01) cardiomyocytes. This shift was reduced, though still significant, when PKC was preceded by PKA, while PKA following PKC did not further decrease pCa(50). Protein analysis indicated that PKC phosphorylated PKA sites in human TnI and increased phosphorylation of troponin T, while myosin light chain phosphorylation remained unaltered. CONCLUSION: In human myocardium PKC-mediated myofilament protein phosphorylation only has a minor effect on maximal force development. The PKC-mediated decrease in Ca(2+) sensitivity may serve to improve diastolic function in failing human myocardium in which PKA-mediated TnI phosphorylation is decreased. PMID- 16376871 TI - Endoplasmic reticulum Ca2+ depletion induces endothelial cell apoptosis independently of caspase-12. AB - OBJECTIVE: Apoptosis of endothelial cells is considered an initial step in the development of atherosclerosis. Recent studies have indicated that depletion of the endoplasmic reticulum (ER) Ca(2+) content plays an important role in apoptosis. Caspase-12 is a key signal in ER stress-induced apoptosis. However, it is not known whether the depletion of ER Ca(2+) is linked to caspase-12 signalling in endothelial cells. Here we have investigated the interaction of Ca(2+) signalling and caspase-12 cleavage in apoptosis of endothelial cells. METHODS: Cytosolic Ca(2+) concentration ([Ca(2+)](i)) of primary porcine aortic endothelial cells was measured using fura-2/AM. Apoptosis was assessed by DNA fragmentation, and cleavage of caspase-12 using Western blotting techniques. RESULTS: Thapsigargin (5 microM), an inhibitor of the ER Ca(2+)-ATPase, depleted ER Ca (2+) content, increased [Ca(2+)](i), cleaved caspase-12, and induced apoptosis. Bradykinin (10 nM) also increased [Ca(2+)](i) but did not cleave caspase-12 or induce apoptosis. However, when intracellular Ca(2+) was chelated with BAPTA/AM (100 microM), bradykinin caused ER Ca(2+) depletion and apoptosis without accompanying caspase-12 cleavage. A non-selective caspase inhibitor, z VAD.fmk (100 microM), inhibited apoptosis and cleavage of caspase-12 stimulated by thapsigargin, while a calpain inhibitor, MDL 28170 (120 microM), inhibited caspase-12 cleavage but not apoptosis. CONCLUSIONS: Thus, increases in intracellular Ca(2+) concentration are not sufficient for the induction of apoptosis in endothelial cells, and ER Ca(2+) depletion appears to induce apoptosis independently of caspase-12. PMID- 16376872 TI - The role of the laminin beta subunit in laminin heterotrimer assembly and basement membrane function and development in C. elegans. AB - Laminins are components of basement membranes that are required for morphogenesis, organizing cell adhesions and cell signaling. Studies have suggested that laminins function as alpha(x) beta(y) gamma(z) heterotrimers in vivo. In C. elegans, there is only one laminin beta gene, suggesting that it is required for all laminin functions. Our analysis is consistent with the role of the laminin beta as a subunit of laminin heterotrimers; the same cells express the laminin alpha, beta, and gamma subunits, the laminin beta subunit localizes to all basement membranes throughout development, and secretion of the beta subunit requires an alpha subunit. RNAi inhibition of the beta subunit gene or of the other subunit genes causes an embryonic lethality phenotype. Furthermore, a distinctive set of phenotypes is caused by both viable laminin alpha and beta partial loss-of-function mutations. These results show developmental roles for the laminin beta subunit, and they provide further genetic evidence for the importance of heterotrimer assembly in vivo. PMID- 16376873 TI - Hemodynamic effects of bupropion in anesthetized dogs. AB - Bupropion is a non-nicotinic drug used in smoking cessation therapy. However, its acute effects remain unclear. In this study, we investigated the effects of bupropion on hemodynamic parameters in pentobarbital-anesthetized mongrel dogs. Bupropion administered either in bolus injections (3 or 6 mg/kg, i.v.) or in cumulative doses of 0.01, 0.1, 1, 3 and 10 mg/kg showed, in both studies, a significant increase of mean pulmonary arterial pressure and pulmonary vascular resistance index. These results show that bupropion can elevate the pulmonary pressure. Further investigations should be done to test this effect in smokers with chronic obstructive pulmonary disease. PMID- 16376874 TI - Cell transplantation to the auditory nerve and cochlear duct. AB - We have developed a technique to deliver cells to the inner ear without injuring the membranes that seal the endolymphatic and perilymphatic chambers. The integrity of these membranes is essential for normal hearing, and the technique should significantly reduce surgical trauma during cell transplantation. Embryonic stem cells transplanted at the internal auditory meatal portion of an atrophic auditory nerve migrated extensively along it. Four-five weeks after transplantation, the cells were found not only throughout the auditory nerve, but also in Rosenthal's canal and the scala media, the most distal portion of the auditory nervous system where the hair cells reside. Migration of the transplanted cells was more extensive following damage to the auditory nerve. In the undamaged nerve, migration was more limited, but the cells showed more signs of neuronal differentiation. This highlights an important balance between tissue damage and the potential for repair. PMID- 16376875 TI - Thr199 phosphorylation targets nucleophosmin to nuclear speckles and represses pre-mRNA processing. AB - Nucleophosmin (NPM) is a multifunctional phosphoprotein, being involved in ribosome assembly, pre-ribosomal RNA processing, DNA duplication, nucleocytoplasmic protein trafficking, and centrosome duplication. NPM is phosphorylated by several kinases, including nuclear kinase II, casein kinase 2, Polo-like kinase 1 and cyclin-dependent kinases (CDK1 and 2), and these phosphorylations modulate the activity and function of NPM. We have previously identified Thr(199) as the major phosphorylation site of NPM mediated by CDK2/cyclin E (and A), and this phosphorylation is involved in the regulation of centrosome duplication. In this study, we further examined the effect of CDK2 mediated phosphorylation of NPM by using the antibody that specifically recognizes NPM phosphorylated on Thr(199). We found that the phospho-Thr(199) NPM localized to dynamic sub-nuclear structures known as nuclear speckles, which are believed to be the sites of storage and/or assembly of pre-mRNA splicing factors. Phosphorylation on Thr(199) by CDK2/cyclin E (and A) targets NPM to nuclear speckles, and enhances the RNA-binding activity of NPM. Moreover, phospho Thr(199) NPM, but not unphosphorylated NPM, effectively represses pre-mRNA splicing. These findings indicate the involvement of NPM in the regulation of pre mRNA processing, and its activity is controlled by CDK2-mediated phosphorylation on Thr(199). PMID- 16376876 TI - A novel strategy to identify the regulatory DNA-organized cooperations among transcription factors. AB - To identify the functional contributions of cooperations among transcription factors on regulatory DNA is critical for understanding transcription activation. But so far there is a great lack of effective identifying methods. Here we describe a novel strategy, based on comprehensively perturbed experiments and a computational model, to identify the cooperations among NF-kappaB (p65), CREB, and AP-1 in transcription activation of human cytomegalovirus major IE1 promoter/enhancer (MIEP). In this strategy, functional profiles of protein-MIEP association and RNA synthesis are achieved through comprehensively perturbing the association of p65, CREB or AP-1 with MIEP and then subjected to the computational model. Consequently, the 'real' cooperations contributing to MIEP activation are found to comprise five but not seven types of potential cooperations. Thus, our research provides a facile systematic approach to identifying the DNA-organized cooperations among transcription factors and understanding transcription activation. PMID- 16376877 TI - In vivo tracing of canonical Wnt signaling in Xenopus tadpoles by means of an inducible transgenic reporter tool. AB - The canonical Wnt pathway is recurrently used during embryogenesis and adult life. To track the cellular output of Wnt signaling in a living organism, we designed a hormone-inducible Wnt responsive system, capable to dynamically and specifically report Wnt pathway activities through eGFP expression. In contrast to previous in vivo reporters, our system essentially avoids interference of consecutive signals by remaining dormant until addition of hormone, which makes it a valuable tool to map canonical Wnt signaling in post-embryonic stages. Transgenic Xenopus laevis embryos were analyzed revealing at tadpole stage in specific tissues and organs cell populations with high Wnt pathway activity. PMID- 16376878 TI - Predicting protein interaction sites from residue spatial sequence profile and evolution rate. AB - This paper proposes a novel method that can predict protein interaction sites in heterocomplexes using residue spatial sequence profile and evolution rate approaches. The former represents the information of multiple sequence alignments while the latter corresponds to a residue's evolutionary conservation score based on a phylogenetic tree. Three predictors using a support vector machines algorithm are constructed to predict whether a surface residue is a part of a protein-protein interface. The efficiency and the effectiveness of our proposed approach is verified by its better prediction performance compared with other models. The study is based on a non-redundant data set of heterodimers consisting of 69 protein chains. PMID- 16376879 TI - Elasticity and adhesion of resting and lipopolysaccharide-stimulated macrophages. AB - Colloidal Force Microscopy was employed to study the viscoelastic and adhesive properties of macrophages upon stimulation with lipopolysaccharide (LPS). Force vs. distance measurements were performed. The adhesion of LPS-stimulated cells (separation force=37+/-3 nN) was almost twice as high as that of resting macrophages (16+/-1 nN). Upon retraction pulling of membrane tethers was observed. Tether lengths and forces at which rupture take place did not depend on stimulation. The reduced Young's modulus K, a measure of cytoskeleton elasticity, was three times lower than that of the control. The data show that LPS has profound effects on cytomechanical and adhesion properties of macrophages. PMID- 16376880 TI - The X protein of hepatitis B virus binds to the F box protein Skp2 and inhibits the ubiquitination and proteasomal degradation of c-Myc. AB - The HBx protein of hepatitis B virus is involved in deregulation of cell cycle and development of hepatocellular carcinoma. Since c-Myc also plays an important role in cell proliferation and tumor development, we studied its regulation by HBx in a human hepatoma cell line. Co-expression of HBx and c-Myc resulted in increased stability of intracellular c-Myc. HBx blocked the ubiquitination of Myc through a direct interaction with the F box region of Skp2 and destabilization of the SCF(Skp2) complex. We suggest that sustained presence of c-Myc combined with mitogenic activity inherent to HBx may be associated with cell cycle deregulation and transformation. PMID- 16376881 TI - ABCA7, a molecule with unknown function. AB - Many ABC proteins are functional in cellular lipid transport in various different manners. ABCA7 is a full-size ABC transporter, the physiological function of which is unknown to date. This is a protein that shows the highest homology known to ABCA1, an essential molecule for producing of plasma high-density lipoprotein (HDL), and in fact it mimics ABCA1 to mediate the production of HDL from cellular lipid when transfected in vitro. It is therefore rational to assume that ABCA7 plays a relevant role in regulating of lipid metabolism. However, the ABCA7 expression profile is distinct from that of ABCA1, with respect to tissue specific distribution and response to some reagents, presumably because of different transcriptional and posttranscriptional regulation. Potential roles and functions of ABCA7 in lipid homeostasis are discussed, especially in relation to HDL metabolism, based on available publications. PMID- 16376883 TI - Gene conversion from SLG to SRK resulting in self-compatibility in Brassica rapa. AB - Self-compatible S-54 homozygotic plants were found in progenies of an F(1) hybrid cultivar in Chinese cabbage. Pollination tests revealed that this self compatibility is controlled by the S locus and caused by the loss of the recognition function of the stigma. SRK, the gene for the recognition molecule in the stigma, was normally transcribed and translated in the self-compatible plants. The 1034-bp region in the receptor domain of SRK in the self-compatible plants was 100% identical to SLG in S-54, while that in self-incompatible S-54 homozygotic plants was 95.1% identical. These results suggest that the self compatibility of the S-54 homozygotes is due to amino-acid changes caused by gene conversion from SLG to SRK. PMID- 16376882 TI - Selective induction of cyclooxygenase-2 plays a role in lysophosphatidic acid regulated Fas ligand cell surface presentation. AB - Previous studies found that lysophosphatidic acid (LPA) upregulated Fas ligand (FasL) presentation on the ovarian cancer cell surface and lead to apoptosis of activated lymphocytes. In this report, we investigated the role of selective induction of cyclooxygenase-2 (Cox-2) in FasL cell surface presentation stimulated by LPA. Ovarian cancer cells pretreated with general aspirin derivative acetylsalicylic acid and specific Cox-2 inhibitor (NS-398) before stimulation with LPA, FasL cell surface presentation was significantly blocked, so was the apoptosis of activated lymphocytes mediated by increasing FasL on the ovarian cancer cell surface. Using the specific inhibitors PD98059, AG1478 or dominant-negative epidermal-growth-factor receptor (EGFR-DN) plasmid, we found that the activation of ERK1/2 played a role in Cox-2 induction, and the transactivation of EGFR worked as an upstream signaling pathway in ERK1/2 phosphorylation. This study first revealed the selective induction of Cox-2 by LPA led to FasL presentation on ovarian cancer cell surface and provide cancer cell immune privilege, and might provide important information of Cox-2 in cancer progression and Cox-2 inhibitors' application in cancer chemoprevention. PMID- 16376884 TI - Characterisation of the interface between nucleophosmin (NPM) and p53: potential role in p53 stabilisation. AB - We have used surface plasmon resonance to quantify the kinetics and stoichiometry of the interaction between p53 and nucleophosmin (NPM). Domains characterising the interface between the two proteins were identified by chemical cross-linking, proteolytic digestion and mass spectrometry based peptide mapping. We show that the C-terminal domain of NPM (residues 242-269) interacts with two regions of p53 (residues 175-196 and residues 343-363) which belong, respectively, to the DNA binding domain and the tetramerisation domain. Potential biological consequences of such interactions are discussed. PMID- 16376885 TI - Arabidopsis KRPs have distinct inhibitory activity toward cyclin D2-associated kinases, including plant-specific B-type cyclin-dependent kinase. AB - Arabidopsis contains seven Kip-related protein (KRP) genes encoding CDK (cyclin dependent kinase) inhibitors (CKIs), which shares a restricted similarity with mammalian p27Kip1. Here, we analyze the characteristics of the KRPs. Although KRP1-KRP7 interact with active cyclin D2 (CYCD2)/CDKA and CYCD2/CDKB complexes to a similar extent, they inhibit kinase activity to a different extent. Our results suggest that inhibitory activity is related to the binding ability between KRP proteins and cyclin/CDK complexes, but secondary and tertiary structure may be also involved. These data provide the first evidence that KRPs inhibit kinase activity associated with plant-specific CDKB. PMID- 16376886 TI - Microarray analysis of gene expression in adult retinal ganglion cells. AB - Retinal ganglion cells (RGCs) transfer visual information to the brain and are known to be susceptible to selective degeneration in various neuropathies such as glaucoma. This selective vulnerability suggests that these highly specialized neurons possess a distinct gene expression profile that becomes altered by neuropathy-associated stresses, which lead to the RGC death. In this study, to identify genes expressed predominantly in adult RGCs, a global transcriptional profile of purified primary RGCs has been compared to that of the whole retina. To avoid alterations of the original gene expression profile by cell culture conditions, we isolated RNA directly from adult RGCs purified by immunopanning without prior sub-cultivation. Genes expressed predominantly in RGCs included: Nrg1, Rgn, 14-3-3 family (Ywhah, Ywhaz, Ywhab), Nrn1, Gap43, Vsnl1, Rgs4. Some of these genes may serve as novel markers for these neurons. Our analysis revealed enrichment in genes controlling the pro-survival pathways in RGCs as compared to other retinal cells. PMID- 16376887 TI - On the contribution of stereochemistry to human ITPK1 specificity: Ins(1,4,5,6)P4 is not a physiologic substrate. AB - Ins(1,4,5,6)P4, a biologically active cell constituent, was recently advocated as a substrate of human Ins(3,4,5,6)P4 1-kinase (hITPK1), because stereochemical factors were believed relatively unimportant to specificity [Miller, G.J., Wilson, M.P., Majerus, P.W. and Hurley, J.H. (2005) Specificity determinants in inositol polyphosphate synthesis: crystal structure of inositol 1,3,4 triphosphate 5/6-kinase. Mol. Cell. 18, 201-212]. Contrarily, we provide three examples of hITPK1 stereospecificity. hITPK1 phosphorylates only the 1-hydroxyl of both Ins(3,5,6)P3 and the meso-compound, Ins(4,5,6)P3. Moreover, hITPK1 has >13,000-fold preference for Ins(3,4,5,6)P4 over its enantiomer, Ins(1,4,5,6)P4. The biological significance of hITPK1 being stereospecific, and not physiologically phosphorylating Ins(1,4,5,6)P4, is reinforced by our demonstrating that Ins(1,4,5,6)P4 is phosphorylated (K(m) = 0.18 microM) by inositolphosphate-multikinase. PMID- 16376888 TI - The SigB sigma factor mediates high-temperature responses in the cyanobacterium Synechocystis sp. PCC6803. AB - The sigma factors of RNA polymerase play central roles when bacteria adapt to different environmental conditions. We studied heat-shock responses in the cyanobacterium Synechocystis sp. PCC6803 using the sigma factor inactivation strains deltasigB, deltasigD and deltasigBD. The SigB factor was found to be important for short-term heat-shock responses and acquired thermotolerance. The normal high-temperature induction of the hspA gene depended on the SigB factor. The SigD sigma factor had a role in high-temperature responses as well, and the double inactivation strain deltasigBD grew more slowly at 43 degrees C than the deltasigB and deltasigD strains. PMID- 16376889 TI - Proteolytic processing and differential distribution of secretogranin-II in goldfish. AB - Secretoneurin (SN) is a 33-34 amino acid neuropeptide derived by endoproteolysis of secretogranin-II (SgII), a chromogranin. A multi-antigenic strategy was used to generate a rabbit polyclonal goldfish SN antiserum that was characterized for Western blot analysis. In the goldfish pituitary two intermediate proteins containing SN and likely processed from the 69.6-kDa SgII precursor were detected. No immunoreactive proteins were observed in the goldfish interrenal, ovary, cerebellum, and telencephalon whereas SgII mRNA was expressed in all these tissues. Immunoreactive levels of the approximately 57 kDa product were higher in the pars distalis (PD) than in the neurointermediate lobe (NIL). The abundance of the approximately 57 kDa protein indicates that this SgII-product containing the SN sequence is a major stored form in secretory granules of the goldfish pituitary. High expression and processing of SN in the hypothalamus and pituitary suggest important roles for SgII-derived peptides in neuroendocrine tissues. PMID- 16376890 TI - Involvement of corticotropin-releasing factor and adrenocorticotropic hormone in the ovarian maturation, seawater acclimation, and induced spawning of Liza ramada. AB - In the present study, we investigated the distribution and activities of corticotropin-releasing factor (CRF) and adrenocorticotropic hormone (ACTH) immunoreactive (ir) cells in the brain and pituitary of Liza ramada during ovarian maturation, seawater acclimation, and induction of spawning. Using immunohistochemistry, we detected that CRF-ir cell bodies exist in different brain regions: medulla oblongata (MO), midbrain tegmentum, habenula, nucleus preopticus (NPO), and in a ventral hypothalamic region corresponding to the nucleus lateralis tuberis (NLTP). In the pituitary gland, we detected some ACTH producing cells in the rostral pars distalis (RPD) containing CRF immunoreactivity. The synthetic and secretory activity of CRF-ir cells in the NPO and MO as well as ACTH-ir cells in the pituitary were enhanced during ovarian maturation. During seawater acclimation, CRF-ir cells in the NPO and MO and ACTH ir cells in the pituitary showed dramatic increases in their synthetic activity. These cells showed dramatic increase in their secretory activity during spawning induced by human chorionic gonadotropin (HCG) injection in L. ramada. Finally, hormonally induced ovulation was accompanied with elevation of plasma cortisol and depletion of CRF and ACTH immunoreactivity within the brain and the pituitary gland, respectively. Taken together, our findings suggest that mature breeders of L. ramada may respond to stress resulting from ovarian maturation, and seawater acclimation as well as induced spawning. Mechanisms include enhancement of the synthetic and/or secretory activity of CRF-ir cells in the NPO and MO as well as ACTH-ir cells in the pituitary gland along with a rise in plasma cortisol during ovulation, supporting the possible role of these hormones during stress and reproduction in L. ramada. PMID- 16376892 TI - The effectiveness of postconditioning and preconditioning on infarct size in hypercholesterolemic and normal anesthetized rabbits. AB - BACKGROUND: Postconditioning with multiple very short coronary occlusions immediately after prolonged ischemia is a new endogenous mechanism for protection of the ischemic heart. We tested whether postconditioning is effective in limiting infarct size in hypercholesterolemic rabbits. METHODS: Male rabbits were fed a cholesterol-enriched diet for 6 weeks. Animals were subjected to 30 min of regional ischemia of the heart and 3h of reperfusion with the following additional interventions: Control group no additional intervention, ischemic preconditioning (IPC) group to two cycles of 5 min ischemia separated by 10 min reperfusion before the index ischemia, 6/10 IPostC group to six cycles of 10s ischemia separated by 10s reperfusion and 4/30 IPostC group to four cycles of 30s ischemia separated by 30s reperfusion immediately after the end of the index ischemia. In a second series of experiments the same interventions were applied to animals fed with a normal diet. Infarct size (I) was determined by triphenyltetrazolium chloride staining. RESULTS: The I/R in hypercholesterolemic rabbits was 55.2+/-5.9% in Control, 17.9+/-4.2% in IPC (P<0.01 versus Control), 63.4+/-4.8% in 6/10 IPostC and 55.6+/-8.2% in 4/30 IPostC (P=NS between them and the Control group). In the normal diet rabbits, the I/R was 48.2+/-4.3% in Control, 15.1+/-2.1% in IPC, 20.4+/-2.9% in 6/10 IPostC (P<0.01 versus Control) and 45.1+/-8.9% in 4/30 IPostC (P=NS versus Control). Blood cholesterol levels were increased and atheromatic lesions were present in the first series. CONCLUSION: Postconditioning is ineffective in limiting the infarct size in rabbits with hypercholesterolemia and atherosclerosis while preconditioning continued to be effective under the same conditions. PMID- 16376893 TI - Treatment with niacin lowers ADMA. PMID- 16376894 TI - Experimental assessment and analytical 2D predictions of the stocking pressures induced on a model leg by Medical Compressive Stockings. AB - Stocking supports have represented, for more than two millennia, the most efficient way to treat the veinous diseases and lymphatic disorders. Although this treatment consists solely in the application of a mechanical pressure to help the blood in reaching back the heart, very little is known on this mechanical effort exerted on a human limb by knitted fabrics. However, nowadays the precise assessment of this pressure distribution is crucial in fitting the treatment to the patient pathology and morphology. In order to describe rationally, for the first time, the pressure distribution induced on a leg, a combined experiment-simulation 2D methodology has been set to validate this mechanical approach. The present article is the first part of a two-papers communication. Experimental aspects are presented here, first to measure these stocking pressures on a rigid leg using the SIGaT((R)) device based on a pneumatic sensor. Then, the knitted fabric mechanical response is characterized under uniaxial tension for large strains, to evaluate the simplified Laplace based pressure that can be compared with the pressure measurements, knowing the local curvature radii of a leg section. This experimental approach is to be completed with numerical simulations of the stocking mechanism on the same model leg. PMID- 16376895 TI - Separation of cationic aracyl derivatives of betaines and related compounds. AB - Cationic aracyl esters of betaines can be formed by alkylation with aracyl halides or trifluoromethanesulfonates. HPLC on a non-endcapped strong cation exchange (SCX) column gave high retention of these derivatives. Cation exchange HPLC may be carried out on a normal-phase (silica or alumina) column using a polar organic solvent (acetonitrile, propan-2-ol) containing an aqueous buffer with an organic cation and a hydrophilic anion. Selectivity is affected by the choice of organic solvent and buffer, e.g. alcohols decrease the retention times of hydroxybetaines such as carnitine. Retention is reduced by increasing the water content and the buffer concentration. Capillary electrophoresis migration times are affected by the choice of buffer anion, with low pH citrate buffers favoured. PMID- 16376896 TI - High-performance liquid chromatography method for simultaneous determination of aliphatic acid, aromatic acid and neutral degradation products in biomass pretreatment hydrolysates. AB - A variety of degradation products are produced upon dilute acid pretreatment of lignocellulosic biomass. Within this larger construct, organic acids, phenols and aromatic aldehydes represent important compound classes to investigate due to increasing evidence of their inhibitory effect on fermentative microorganisms. An analytical extraction procedure is presented, enabling isolation of potential analytes away from alternative products in biomass hydrolysates. Additionally, a reversed-phase high-performance liquid chromatography (HPLC) method has been developed and validated, affording simultaneous separation and quantitative determination of 32 potential analytes in water with UV detection at 210 nm. The method was subsequently employed to quantify a variety of aliphatic acid, aromatic acid, aldehyde and phenolic degradation products in a corn-stover hydrolysate at concentration levels ranging from 0.02 to 41 mM. PMID- 16376897 TI - Molecularly imprinted polymers grafted to flow through poly(trimethylolpropane trimethacrylate) monoliths for capillary-based solid-phase extraction. AB - Monolithic molecularly imprinted polymers (mMIPs) have been synthesized in a novel way using a trimethylolpropane trimethacrylate core material photo polymerized in situ in a 100 microm I.D. UV-transparent capillary and further photo-grafted to create specific cavities in the grafted layer. This polymerization technique allows the imprints to be directly created on the surface of the material using a minimum amount of template. Three different anaesthetics of similar structures (bupivacaine, mepivacaine and S-ropivacaine) were used as model target molecules to synthesize sample enrichment media. Hence, various mMIPs have been prepared and evaluated on a micro-system against each analyte in order to test the retention properties and cross-selectivities of the materials. The retention factors were determined and compared with the non imprinted reference column (mNIP), yielding high imprinting factors together with good selectivity factors between the three analytes. A study with a pure enantiomeric target was carried out to assess the degree of stereo-specific imprinting for injection of racemic mixtures. Finally, one column was imprinted with an equimolar mixture of all three anaesthetics to provide further comprehension of the retention mechanism and accredit the possibility of using the material as a sample enrichment entity. Scanning electron microscopy (SEM), nitrogen absorption/desorption (BET) and mercury intrusion porosimetry were used to characterize the monolith and the mMIPs properties. Nuclear magnetic resonance (NMR) has been used to assess the similarities between the mMIP and mNIP. PMID- 16376898 TI - Micellar liquid chromatography retention model based on mass-action concept of micelle formation. AB - Mass-action model of surfactant micelle formation has been used to develop a conceptual retention model in micellar liquid chromatography (MLC). The retention model bases on the consideration of the changes of the sorbate microenvironment at its transferring from the mobile phase (hybrid micellar eluent) to the stationary phase (a modified surface of alkyl-bounded sorbent). Principal retention equation contains the characteristics of hybrid micelles (critical micelle concentration, degree of counterion binding, partition coefficient of modifier between aqueous solution and micellar pseudo-phase) as well as three fitting parameters. The fitting parameters are an absolute term and coefficients that are equal to the number of molecules of surfactant and modifier, which are attached/detached by sorbate transferring from a hybrid micellar eluent to a modified surface of the stationary phase. On the MLC separation of five antibiotics of rubomicin derivatives and four esters of 4-hydroxybenzoic acid the model of the change of sorbate microenvironment has been tested. The adequateness of model to experimental data has been shown. A simple three-parameter function connecting log k with log cS and log cR that provides a high goodness-of-fit follows from principal retention equation (cS and cR are the molar concentrations of surfactant and organic modifier in the micellar eluent, respectively). PMID- 16376899 TI - Rapid analysis of oligosaccharides derived from glycoproteins by microchip electrophoresis. AB - A novel method for fast profiling of complex oligosaccharides released from glycoproteins based on microchip electrophoresis (mu-CE) is presented here. The characterization of separation conditions, i.e., the composition, concentration and pH of running buffer as well as the applied voltage, has been performed using maltose (G2), cellobiose ( G2'), maltriose (G3) and panose (G3') as oligosaccharide isomer models. In mu-CE, much better separation of oligosaccharide isomers and oligosaccharide ladder was obtained in phosphate buffer than in borate buffer over a wide pH range. Under optimal conditions, high performance separation of the N-linked complex oligosaccharides released from ribonuclease B, fetuin, alpha1-acid glycoprotein (AGP) and IgG was achieved using polymethylmethacrylate (PMMA) microchips with an effective separation channel of 30 mm. These results represent the first reported analysis of the N-linked oligosaccharides derived from glycoproteins by mu-CE, indicating that the present mu-CE-based method is a promising alternative for characterization of the N linked oligosaccharides in glycoproteins. PMID- 16376900 TI - In vivo protein sampling using capillary ultrafiltration semi-permeable hollow fiber and protein identification via mass spectrometry-based proteomics. AB - Here, we advanced a novel technique using capillary ultrafiltration (CUF) probes to collect in vivo secreted proteins in the subcutaneous tissue of mouse ear. We fabricated two kinds of CUF probe, one with and one without a semi-permeable membrane hollow fiber. Proteins collected by CUF probes were profiled and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MADLI-TOF-MS) and quadrupole time-of-flight tandem mass spectrometry (Q-TOF-MS/MS) without using two-dimensional gel electrophoresis (2 DE) separation. Five proteins including cofilin-1, futuin-A, complement C3, gelsolin, and apolipoprotein C-1 were identified from the sample collected by the CUF probe with a semi-permeable membrane hollow fiber. The presence of well documented secretory proteins supports the efficiency of CUF probes in sampling in vivo secreted proteins. We also found that hemoglobin collected by the CUF probe without a semi-permeable membrane hollow fiber completely masked protein identification by mass spectrometry. The presence of relatively large amounts of hemoglobin in this condition illustrates the necessity of the semi-permeable membrane hollow fiber to the technique of CUF probe in conjunction with mass spectrometry. Also, the technique represents a powerful method for the identification of in vivo secreted proteins and has potential application for in the detection of biomarkers for human diseases. PMID- 16376901 TI - Chiral capillary electrophoretic analysis of verapamil metabolism by cytochrome P450 3A4. AB - Cytochrome P450 (CYP), which is one of the most important enzymes in human liver, is responsible for a large portion of the first-pass metabolism of drugs. Many studies have focused on the determination of CYP activity by substrate assays. Most of them used liquid chromatography (LC) as analytical technique, while only a few studies used capillary electrophoresis (CE) for the separation and quantitation of reaction components. In this study, the feasibility of using CE in an in vitro metabolism study with CYP was tested. Verapamil was chosen as the substrate for CYP 3A4 isozyme (Supersome). A chiral capillary electrophoretic method was developed and validated for the simultaneous determination of R,S verapamil (VER) and their major metabolites, R,S-norverapamil (NOR). A method for CYP 3A4 activity assay was proposed with VER as a probe. At the same time, the enantioselective metabolism of VER was studied. Michaelis-Menten constants of R- and S-VER were determined. S-VER was metabolised faster and more extensively than R-VER, with K(m)=167+/-23 microM, V(max)=3,418+/-234 pmol/min/mg for S-VER, and K(m)=168+/-35 microM, V(max)=2,502+/-275 pmol/min/mg for R-VER. PMID- 16376902 TI - Measurement of monomolecular binding constants of neutral phenols into the beta cyclodextrin by continuous frontal analysis in capillary and microchip electrophoresis via a competitive assay. AB - Measurement of binding constant by chip electrophoresis is a very promising technique for the high throughput screening of non-covalent interactions. Among the different electrophoretic methods available that yield the binding parameters, continuous frontal analysis is the most appropriate for a transposition from capillary electrophoresis (CE) to microchip electrophoresis. Implementation of this methodology in microchip was exemplified by the measurement of inclusion constants of 2-naphtalenesulfonate and neutral phenols (phenol, 4-chlorophenol and 4-nitrophenol) into beta-cyclodextrin by competitive assays. The issue of competitor choice is discussed in relation to its appropriateness for proper monitoring of the interaction. PMID- 16376903 TI - Continuous microwave-assisted extraction coupled on-line with liquid-liquid extraction: determination of aliphatic hydrocarbons in soil and sediments. AB - This paper describes a new extraction method for the determination of aliphatic hydrocarbons (AHs) in soil and sediment samples, using continuous microwave assisted extraction (MAE) combined with liquid-liquid extraction, for clean-up purposes. Analytical determinations were carried out by gas chromatography coupled with impact ionization mass spectrometry. The influence of the experimental conditions was tested using an agricultural soil spiked with standards (stored at 4 degrees C for 1 month) as reference soil. Maximum extraction efficiencies (80-90%) were achieved using 0.1-1.0g of sample, 60microl of water and 3ml of n-hexane (extractant) and 5min of extraction time; less than 70% of the most volatile hydrocarbons (C(9)-C(12)) were recovered since many evaporated during the drying step of the sample. MAE was compared with a conventional extraction method such as Soxhlet and a good agreement in the results was obtained (average recovery percentage value of 105% by comparing MAE against Soxhlet). Quality parameters such as linear range (0.5-800microg/g), limits of detection (LODs) (0.1-0.2microg/g) and precision (RSD, 4-6%) were determined using spiked soil samples. This method was successfully applied to the analysis of aliphatic hydrocarbons (C(9)-C(27) including pristane and phytane) in contaminated real samples. PMID- 16376904 TI - Looking inside the pores of a chromatographic column. I. Variation of the pore volume with mobile phase composition. AB - The dependence of void volume and pore volume on mobile phase composition was studied on a reversed phase column based on poly(divinyl benzene) in mobile phases consisting of 100% tetrahydrofuran (THF) and 40-90% THF-water. Void volumes V0 are obtained from a series of polymers with different molar mass by extrapolation of elution volume to M = 0, the interstitial volume Vi from the elution volume of polymers with a sufficiently high molar mass. The pore volume Vp is obtained as the difference (V0 - Vi). While Vi does not depend on mobile phase composition, a dramatic decrease of the pore volume is observed, when the mobile phase is changed from THF to THF-water. This effect of the mobile phase composition on the pore volume is observed using PEG and PPG as test macromolecules, regardless the retention mechanism: at 40-50% THF both PEG (in exclusion mode) and PPG (in adsorption mode) experience the same reduced pore volume, which is considerably lower than that in 100% THF. PMID- 16376905 TI - Determination of bisphenol diglycidyl ether residues in canned foods by pressurized liquid extraction and liquid chromatography-tandem mass spectrometry. AB - A new confirmatory method for simultaneous determination of bisphenol diglycidyl ether residues (BADGE, BADGE.H(2)O, BADGE.2H(2)O, BADGE.H(2)O.HCl, BADGE.HCl, BADGE.2HCl, BFDGE and BFDGE.2HCl) from canned food has been developed. The proposed method includes extraction by pressurized liquid extraction (PLE) followed by liquid-liquid partition and purification by solid phase extraction (SPE). Several solvent systems and different operating conditions (time, temperature) have been investigated for PLE optimization. A reversed-phase high performance liquid chromatography (RP-HPLC) coupled to atmospheric pressure chemical ionisation tandem mass spectrometry (APCI-MS-MS) method was developed for the separation, quantification and confirmation. The ion source settings were optimized using a design of experiments (DOE). The optimized method was applied to the determination of these chemicals at very low levels in different samples with a quantification limit of 5 ng/g. Recoveries ranged between 82 and 101% and standard deviations were less than 10%. PMID- 16376906 TI - Low level determination of p-toluenesulfonate and benzenesulfonate esters in drug substance by high performance liquid chromatography/mass spectrometry. AB - GC/FID and HPLC/MS single ion monitoring methods have been evaluated for the determination of trace levels of methyl, ethyl and isopropyl esters of p toluenesulfonic acid and methyl, ethyl, isopropyl and n-butyl esters of benzenesulfonic acid in drug substances. These sulfonate esters have been highlighted as potential genotoxins. HPLC/MS was found to be more promising and limits of quantification were between 2.5 and 5 ng/mL, which enabled detection limits in drug substance at 0.01-0.1 ppm for a 50 mg/mL solution. For one drug substance excellent recoveries of 94-95% were obtained at the 1.0 ppm level, however, with a second drug substance, a besylate salt, recoveries ranged from 86% to 100% and were dependant on the sample preparation. Limited stability of the sulfonate esters in various potential sample solutions indicated that samples may need to be prepared immediately before injection. PMID- 16376907 TI - A protocol for designing comprehensive two-dimensional liquid chromatography separation systems. AB - In this paper a protocol is proposed for establishing suitable column dimensions (length and diameters), particle sizes, flow rates, and second-dimension injection volumes (i.e. loop sizes) in comprehensive two-dimensional liquid chromatography (LC x LC). The chromatographer should select the maximum allowable first-dimension retention time, which is approximately equal to the overall analysis time. Also, (s)he should define the maximum allowable pressure in both dimensions and the (minimum) diameter of the first-dimension column. The proposed protocol provides design parameters corresponding to the ideal (theoretically optimal) conditions or to realistic practical conditions. The protocol also allowed us to study the implications of contemporary developments in LC, such as the use of high temperatures (implying reduced viscosities and increased diffusion coefficients), monolithic columns (implying smaller flow-resistance factors), and ultra-high-pressure LC. The combination of (reversed-phase or normal-phase) liquid chromatography with size-exclusion chromatography (LC x SEC) is frequently employed for analysing complex polymers. The proposed protocol is used to design a suitable LC x SEC system under realistic conditions. The results resemble the systems that have been designed and implemented by expert chromatographers, but they also indicate how current practice can be improved. PMID- 16376908 TI - On-line cut-off technique and organic modifier addition aided signal enhancement for trace analysis of carbohydrates in cellulase hydrolysate by ion exclusion chromatography-electrospray ionization mass spectrometry. AB - Paper cellulose has been hydrolyzed with calcium alginate immobilized cellulase to produce carbohydrate products and the three trace sugars, galactose, arabinose, and mannose in the cellulase hydrolysate have been analyzed by HPIEC/ESI-MS. Applying the on-line cut-off technique to the HPIEC/ESI-MS can cut the high concentration glucose off to eliminate its interference on the peaks of minor sugars and enhance their signals from 1.1- to 1.6-fold. However, the on line post column addition of 15% ethanol to the eluate can increase the signal of the three trace sugars, galactose, arabinose, and mannose up to 17-, 23-, and 11 fold, respectively, and make the corresponding detection limits as 0.04, 0.04, and 0.03 ppm. The accuracies of the quantitative analysis for the three trace sugars with the signal enhanced HPIEC/ESI-MS by the two enhancement methods were larger than 95%. The precisions of the analytical results were also greatly improved by the assistance of the two techniques and were less than 6.5%. The quantitative analysis of the three trace sugars was performed with the internal standard method and the internal standard (IS) was sorbitol. Overall, the signal enhancement of HPIEC/ESI-MS and quantification of the three trace sugars by the on-line cut-off technique and organic modifier addition was successful. PMID- 16376909 TI - Simple analysis of four bisphosphonates simultaneously by reverse phase liquid chromatography using n-amylamine as volatile ion-pairing agent. AB - Volatile organic amine was used as the mobile phase addictive during the separation of four bisphosphonates (alendronate, pamidronate, zoledronic acid and etidronate). An isocratic liquid chromatography method with evaporative light scattering detection (ELSD) was developed for these bisphosphonates which are not retained on non-polar column and lack chromophore for detection. The analytes have sufficiently separated from each other on a Phenomenex C18 column. The effects of mobile phase composition and instrumental parameters of ELSD were studied. This newly developed method enables direct measurement for analysis of bisphosphonates without the need of derivatization. This developed method provides high separation and specificity to bisphosphonate analysis. In quantitative analysis, the method showed satisfactory precision (less than 2.8%) and accuracy (higher than 94.4%), good linearity (r=0.9991-0.9997) and sufficient sensitivity (15-18 microg/ml). It can be easily and conveniently adopted for the routine quality control analysis. PMID- 16376910 TI - Determination of ergosterol in Fusarium-infected wheat by liquid chromatography atmospheric pressure photoionization mass spectrometry. AB - A rapid liquid chromatography-atmospheric pressure photoionization mass spectrometry (LC-APPI-MS) method was developed for the determination of ergosterol in wheat grains. The effects of the dopants acetone, toluene and anisole on the ionization efficiency were studied. To identify the predominant ions, APPI-MS-MS studies were performed. Different LC and MS parameters were optimized to obtain maximum sensitivity. The effects of the mobile phase composition and of the flow rate were investigated. Additionally, the effects of the nebulizer gas pressure, the drying gas flow, the vaporizer temperature, the fragmentor voltage and the capillary voltage on the ionization efficiency were evaluated. The calibration curve exhibited good linearity and reproducibility. The detection limit (S/N=3) was 0.15 ng on column, which allows the determination of ergosterol in wheat at a concentration as low as 0.12 microg/g. Twenty wheat varieties artificially infected with Fusarium graminearum were investigated by this method. PMID- 16376911 TI - Detection and quantification of low-molecular-weight aldehydes in pharmaceutical excipients by headspace gas chromatography. AB - The adverse effect of reactive chemical residues on the quality of drug products has necessitated the determination of low-molecular-weight aldehydes in pharmaceutical excipients. An analytical methodology for the detection of trace amounts of C1-C8 aliphatic aldehydes and benzaldehyde in excipients is described. The proposed procedure is based on the derivatization of aldehydes in aqueous solution with O-2,3,4,5,6-(pentafluorobenzyl) hydroxylamine hydrochloride (PFBHA), followed by static headspace gas chromatographic (SHS-GC) analysis of PFBHA aldehyde oximes with negative chemical ionization (NCI) MS detection. The method developed was demonstrated to be simple, selective, sensitive, and was successfully applied to the screening of aldehydes at sub-microg/g levels in over 30 typical excipients. The most abundant aldehydes found in the samples were formaldehyde and acetaldehyde, for which a rapid and reliable routine quantification method by readily available SHS-GC instrumentation coupled with flame-ionization detection was also developed and validated. PMID- 16376912 TI - Characterisation by liquid chromatography-electrospray tandem mass spectrometry of anthocyanins in extracts of Myrtus communis L. berries used for the preparation of myrtle liqueur. AB - Anthocyanins in extracts of berries of Myrtus communis, prepared following a typical Sardinia myrtle liqueur recipe, were identified and quantified by HPLC coupled with electrospray/tandem mass spectrometry using, respectively, an ion trap and a triple quadrupole mass analyser. The fragmentation patterns of the anthocyanidins were dependent on the MS technique employed, and differed considerably from those previously reported. The anthocyanin profile of five anthocyanin glucosides and four anthocyanin arabinosides, the latter not previously identified in this specie, was specific for myrtle berry extracts. The quantitative compositions of extracts of myrtle berries derived from different geographical areas in Italy were compared. PMID- 16376913 TI - Simultaneous quantification of reduced and oxidized glutathione in plasma using a two-dimensional chromatographic system with parallel porous graphitized carbon columns coupled with fluorescence and coulometric electrochemical detection. AB - A method for the simultaneous quantification of reduced and oxidized glutathione in human plasma employing a two-dimensional chromatographic system with parallel porous graphitized carbon (PGC) columns coupled with fluorescence (FLD) and coulometric electrochemical detection (ED) has been developed. Post-sampling oxidation of reduced glutathione (GSH) was prevented by derivatizing the -SH group with monobromobimane (MBB) and the glutathione-bimane adduct (GSMB) was detected by FLD. Oxidized glutathione (GSSG) was detected by ED optimized to give lowest possible limits of detection (LOD). The method is fully validated and is currently used for determination of GSH, GSSG and its redox potential in different clinical studies. PMID- 16376914 TI - Solid-phase microextraction with on-fiber silylation for simultaneous determinations of endocrine disrupting chemicals and steroid hormones by gas chromatography-mass spectrometry. AB - Based on solid-phase microextraction (SPME) and on-fiber silylation, a method for simultaneous determinations of exogenous endocrine disrupting chemicals (EDCs) and endogenous steroid hormones in environmental aqueous and biological samples by gas chromatography-mass spectrometry (GC-MS) was developed. The selected target compounds were: octylphenol (OP), technical grade nonylphenol (t-NP), diethylstilbestrol (DES), dehydroisoandrosterone (DEHA), estrone (E1), 17beta estradiol (E2), testosterone (T) and pregnenolone (PREG). The optimization of operating conditions influencing the performances of SPME and derivatization were studied in detail. The average correlation coefficient of the calibration curves of the target compounds was 0.9968 and the linear ranges of most compounds spanned over three orders of magnitude. The LOD/(LOQ) values of the target compounds in river water and blood serum were in the range of 0.002-0.378/(0.008 1.261) microg L(-1) and 0.004-0.474/(0.013-1.579) microg L(-1), respectively, which were a bit higher than those in the pure water due to matrix effects. The developed method was applied to the determinations of target compounds in real samples. Exogenous OP, t-NP and DES were at 0.15, 4.67 and 0.02 microg L(-1) in river water and 3.21, 12.17 and 0.15 microg L(-1) in fish blood serum. Natural steroid hormones E1, E2 and T were at 0.18, 0.10 and 5.55 microg L(-1) in river water, and in female fish serum, E1, E2 and PREG were at 1.61, 1.08 and 4.58 microg L(-1), respectively. The proposed SPME method was compared with traditional SPE procedure and the results found using both methods were in the same order of magnitude and both are quite agreeable. PMID- 16376915 TI - Studies on the interaction of some azo dyes (naphthol red-J and direct orange) with nontronite mineral. AB - The adsorption of two azo dyes, namely naphthol red-J and direct orange, on nontronite mineral was studied as a function of pH and temperature. All the sorption studies were conducted in batch mode. The Langmuir and Freundlich isotherm models were applied. The isotherms are Langmuirian in nature, while the Freundlich equation is only valid over the low concentration range. The adsorption of the two dyes increases with increased temperature and decreases with increased pH. The data have been explained in terms of the area of contact of the dye molecule on the clay platelet vis-a-vis the area per anion exchange site. Estimated area per anion exchange site of the mineral and area associated per anionic dye molecule at nontronite surface was 642 and 454 A2 for naphthol red-J and 642 and 440 A2 for direct orange respectively. This has been further supported by X-ray measurements. Thermodynamic parameters of the process were evaluated. PMID- 16376916 TI - Chemical activation of carbon mesophase pitches. AB - This paper studies the chemical activation of mesophase pitches of different origins in order to obtain activated carbons suitable for use as electrodes in supercapacitors. The effect that the activating agent (NaOH, LiOH, and KOH), the alkaline hydroxide/pitch ratio, and the activation temperature had on the characteristics of the resultant activated carbons was studied. LiOH was found to be a noneffective activating agent, while activation with NaOH and KOH yielded activated carbons with high apparent surface areas and pore volumes. The increase of the KOH/pitch ratio caused an increase of the chemical attack on the carbon, producing higher burnoffs and development of porosity. Extremely high apparent surface areas were obtained when the petroleum pitch was activated with 5:1 KOH/carbon ratio. The increase of the activation temperature caused an increase of the burnoff, although the differences were not as significant as those derived from the use of different proportions of activating agent. PMID- 16376917 TI - Interaction of cationic surfactants with carboxymethylcellulose in aqueous media. AB - We have examined the polymer-surfactant interaction in mixed solutions of the cationic surfactants, i.e., dodecyltrimethylammonium chloride, dodecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, tetradecyltriphenylphosphonium bromide, and tetradecylpyridinium bromide and a semiflexible anionic polyelectrolyte carboxymethylcellulose in water and aqueous salt solutions by various techniques: tensiometry, viscosimetry or ion-selective electrode method, and dynamic light scattering. We have investigated the effect of varying surfactant chain length, head group size, counterion, and ionic strength on the critical aggregation concentration (CAC) of mixed polymer surfactant systems and the collapse of the polymer molecule under different solution conditions. The CAC decreases with increasing alkyl chain length. Above a certain surfactant concentration, mixed aggregates start growing until their macroscopic phase separation. The growth is more rapid with greater surfactant tail length and with increasing head group size. This is attributed in both cases to the increasing hydrophobic interaction between polymer and surfactant. Among surfactants with monovalent halide counterions, iodide induces the strongest binding, reflected by the onset of growth of the mixed aggregates at low surfactant concentration. This is perhaps related to the decreasing hydration of the counterion from chloride to iodide. The surfactant concentration at which the viscosity of the solution starts to decrease sharply is smaller than the CAC, and probably reflects polymer chain shrinkage due to noncooperative binding. PMID- 16376918 TI - Thermodynamics of monochlorophenol isomers and pyrite interfacial interactions in the activation state. AB - Thermodynamic parameters of the activation state for phenol and three monochlorophenol (MCP) isomer-pyrite complexes, i.e., MCP isomers used were 2 chlorophenol (2-CP), 3-chlorophenol (3-CP), 4-chlorophenol (4-CP), have been derived from the temperature-dependent kinetic data. Both the initial rate and adsorption density values increased in the order phenol < 2-CP < 3-CP < 4-CP. This suggests that the presence of chlorine substituent on the aromatic ring results in enhanced CP adsorption on pyrite. The activation energy (E(a)), Gibbs free energy (deltaG#), entropy (deltaS#), and enthalpy (deltaH#) of the activation stage for MCP adsorption on pyrite were calculated by Arrhenius and Eyring models. Always deltaS# values approximate to zero and -TdeltaS# values are positive, which indicates that the activation state of MCP adsorption process is entropy-controlled, and the observed linear dependence of deltaH# on -TdeltaS# signals an entropy-enthalpy compensation effect of the MCP adsorption process. The gammaMCP data were quantified well both by 1 - pK diffused double layer (1 - pK DLM) and Langmuir models. PMID- 16376919 TI - High internal phase CO2-in-water emulsions stabilized with a branched nonionic hydrocarbon surfactant. AB - A nonionic-methylated branched hydrocarbon surfactant, octa(ethylene glycol) 2,6,8-trimethyl-4-nonyl ether (5b-C12E8) emulsifies up to 90% CO2 in water with polyhedral cells smaller than 10 microm, as characterized by optical microscopy. The stability of these concentrated CO2/water (C/W) emulsions increases with pressure and in some cases exceeds 24 h. An increase in pressure weakens the attractive van der Waals interactions between the CO2 cells across water and raises the disjoining pressure. It also enhances the solution of the surfactant tail and drives the surfactant from water towards the water-CO2 interface, as characterized by the change in emulsion phase behavior and the decrease in interfacial tension (gamma) to 2.1 mN/m. As the surfactant adsorption increases, the greater tendency for ion adsorption is likely to increase the electrostatic repulsion in the thin lamellae and raise the disjoining pressure. As pressure increases, the increase in disjoining pressure and decrease in the capillary pressure (due to the decrease in gamma) each favor greater stability of the lamellae against rupture. The electrical conductivity is predicted successfully as a function of Bruggeman's model for concentrated emulsions. Significant differences in the stability are observed for concentrated C/W emulsions at elevated pressure versus air/W or C/W foams at atmospheric pressure. PMID- 16376920 TI - Photophysics and charge dynamics of Q-PbS based mixed ZnS/PbS and PbS/ZnS semiconductor nanoparticles. AB - The surface of ZnS and PbS has been modified by interfacing PbS on ZnS and ZnS on PbS nanoparticles. This produced core-shell nanocomposites ZnS/PbS and PbS/ZnS with tunable electronic properties. In both structures PbS particles are present in cubic form with an average diameter of about 6 nm. The addition of Pb2+ (3 x 10(-4) mol dm(-3)) to Q-ZnS (1.5 x 10(-4) mol dm(-3)) in the basic pH range produces size-quantized fluorescent PbS particles coated by metal hydroxides. In these particles the relaxation kinetics of charge carriers has been followed using a picosecond single-photon counting technique. At 1.5 x 10(-4) mol dm(-3) Pb2+ an interfacial relaxation of charge from ZnS to PbS phase could be observed in subnanosecond time domain. An increase in [Pb2+] from 2 x 10(-4) to 1 x 10(-3) mol dm(-3) enhanced the average emission lifetime from 9.4 to 19.4 ns. Composite PbS/ZnS particles are produced at high [ZnS] only. These particles had emission lifetime in mus time range. The extent of charge separation and the dynamics of charge carriers could be manipulated by the surface modification of these nanostructures. PMID- 16376921 TI - Removal of pentachlorophenol from aqueous solutions by dolomitic sorbents. AB - The partial decomposition of dolomite carried out within the temperature range 600-1000 degrees C provides new sorbents, called dolomitic sorbents. Their surface properties and identification by X-ray diffraction are discussed. The lowest specific surface area value was found for the raw dolomite, while the highest value was achieved by the D-1000 sample. The adsorption equilibrium of pentachlorophenol from aqueous solutions on the examined sorbents was investigated at 30, 40, and 50 degrees C via a bath process. Langmuir, Freundlich, Langmuir-Freundlich, and Redlich-Petersen models were fitted to experimental equilibrium data, and their goodnesses of fit are compared. The adjustable parameters of Langmuir-Freundlich and Redlich-Petersen isotherms were estimated by nonlinear least-squares analysis. Langmuir and Freundlich models were found insufficient to explain the adsorption equilibrium, while Langmuir Freundlich and Redlich-Petersen isotherms provide the best correlation of the pentachlorophenol adsorption onto dolomitic sorbents. PMID- 16376922 TI - Dechlorination of beta-methylallyl chloride by electrogenerated [Co(I)(bipyridine)3]+: an electrochemical study in the presence of cationic surfactants. AB - We have investigated the electrocatalytic dehalogenation of beta-methylallyl chloride (beta-mAC), widely used in the polymer industry, using [Co(I)(bpy)3]+ (where bpy=2,2'-bipyridine) electrochemically generated in situ from [Co(II)(bpy)3]2+ at a glassy carbon electrode in the presence of three different cationic surfactants in aqueous solution. Cetyltrimethylammonium bromide (CTAB), tetradecyltrimethylammonium bromide (TDTAB), and cetylbenzyldimethylammonium chloride (CBDAC) were employed in the present investigation. The [Co(II)(bpy)3]2+ cationic surfactant systems show excellent electrocatalytic activity toward dehalogenation of beta-mAC. The dependence of the catalytic current, the corresponding potential, and the current function on the potential scan rate has been analyzed to assess the nature of the catalytic reaction. The second-order rate constant, kchem, for the reaction between the beta-mAC substrate and the electrogenerated-micelle stabilized-Co(I) complex has been calculated by a cyclic voltammetry technique. The reduction products after 3 h of bulk electrolysis have been identified by GC/MS to be one nonchloro compound (2-methyl-1,5-hexadiene (IV)) and two chloro compounds (1-chloro-2,5-dimethyl-2,5-hexadiene (V) and spiro[1.2]cylopropyl-6-chloro-5-methyl-hex-4-ene (VI)). Based on the electrochemical results and the mass spectral data, a reaction scheme involving all the reduction products has been proposed. Finally, a good correlation between the catalytic efficiency and the structural features of the surfactant molecules is demonstrated. The present study emphasizes the need for further optimization work to achieve maximum yield of nonchloro compound (IV) to employ the present [Co(II)(bpy)3]2+-cationic surfactant systems with a high catalytic efficiency as promising for possible applications. PMID- 16376923 TI - The role of few-asperity contacts in adhesion. AB - The surface roughness of a few asperities and their influence on the work of adhesion is of scientific interest. Macroscale and nanoscale adhesion data have seemingly given inconsistent results. Despite the importance of bridging the gap between the two regimes, little experimental work has been done, presumably due to the difficulty of the experiment needed to determine how small amounts of surface roughness might influence adhesion data lying in between the two scales. To investigate the role of few-asperity contacts in adhesion, the pull-off force was measured between different sized atomic-force microscope (AFM) tips (with different roughnesses) and sample surfaces that had well-controlled material properties. There were seventeen tips of four different types, with radii from 200 nm to 60 microm. The samples were unpatterned single crystal silicon with a chemical silicon dioxide surface resulting from a standard silicon wafer clean. Some of the samples were treated with a few angstroms of vapor deposited diphenylsiloxane. We observed that the uncorrected (for surface roughness) pull off force was independent of the radius of the AFM tip, which was contrary to all continuum-mechanics model predictions. To explain this behavior, we assumed that the interactions between the AFM tip and sample were additive, material properties were constant, and that the AFM tip, asperities, and sample surfaces were of uniform density. Based on these assumptions, we calculated a simple correction due to the measured root mean square (RMS) surface roughness of the AFM tips. The simple correction for the RMS surface roughness resulted in the expected dependence of the pull-off force on radius, but the magnitudes were higher than expected. Commercial and heat-treated AFM tips have minimal surface roughness and result in magnitudes that are more reliable. The relative uncertainty for the pull-off force was estimated to be 10%. In this paper, we derive how the cantilever and tip parameters contribute to the measured pull-off force and show how the corrected results compare with theory. Although much work is still needed, the work presented here should advance the understanding of adhesion between the macroscale and nanoscale regimes. PMID- 16376924 TI - Fundamental studies on the intermediate layer of a bipolar membrane V. Effect of silver halide and its dope in gelatin on water dissociation at the interface of a bipolar membrane. AB - The effect of silver ions on the water dissociation of bipolar membranes was first investigated in this paper. To do this, the bipolar membranes were prepared by immersing the anion exchange layers in an AgNO3 solution and then coating a solution of sulfonated polyphenylene oxide (SPPO) on the anion exchange layers. XPS and AES observations indicated that silver at the intermediate layer was in the form of AgCl. The experimental results proved that AgCl has an excellent catalytic function for water dissociation in terms of I-V curves, and the quantity of AgCl played an important role in the behavior of a bipolar membrane. The bipolar membranes with gelatin and the gelatin doped with silver as a catalytic layer were also prepared in the same way, and their I-V behavior and the water dissociation pilot tests were also investigated. The experimental results showed that in the case of gelatin alone, the voltage drop increased slightly at high gelatin concentrations, due mainly to the steric effect and electrostatic interaction, but decreased at low gelatin concentrations due to the hydrophilicity. However, when gelatin was doped with AgCl, the bipolar membranes have an appreciable improvement in both stability and catalytic function, in comparison with those prepared from silver or gelatin. PMID- 16376925 TI - Solubilization and location of phenethylalcohol, benzaldehyde, and limonene in lamellar liquid crystal formed with block copolymer and water. AB - Three fragrances, phenethylalcohol, benzaldehyde, and limonene (that differ in their polarity), were solubilized in lamellar liquid crystal using the polyethylene oxide-polypropylene oxide-polyethylene oxide, (EO27PO61EO27), P104 triblock copolymer, and water. The interlayer spacing of the lamellar liquid crystal were established using small angle X-ray scattering (SAXS). The SAXS data are used to calculate the interfacial area per polyethylene oxide (PEO) block and to determine the location of each fragrance in the association structure. The results show the presence of phenethylalcohol at the interface while part of benzaldehyde contributing to the interface and the other part being located inside the polar/apolar domains. The third fragrance, limonene, did not participate in the formation of the interface and is located inside the apolar domain. PMID- 16376926 TI - Self-assembly of Ag nanoparticle-biotin composites into long fiberlike microstructures. AB - An end-to-end assembly of spherical Ag nanoparticles takes place in the presence of biotin to form long fiberlike microstructures. These microstructures are about 4 mum long with a thickness of 1 mum, obtained from SEM studies. TEM studies showed the presence of spherical silver nanoparticles having an average size of 20 nm. ATR-FTIR studies revealed that silver ions interact with biotin involving the carboxylate group. A weak binding of the silver particles with the thioether and ureido groups helps in connecting the Ag nanoparticles to form long fiberlike structures. Elucidation of the mechanism of formation of the spherical Ag clusters was done by pulse radiolysis. PMID- 16376927 TI - Adsolubilization of 2-naphthol into adsorbed layer of PEO-PPO-PEO triblock copolymers on hydrophilic silica. AB - Adsolubilization of 2-naphthol into an adsorbed layer of poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) triblock copolymers (Pluronics) on hydrophilic silica has been investigated. Four kinds of Pluronics (P103, P105, P123, and F108) were used in order to understand the effect of the hydrophobicity of surfactant on the adsolubilization. The order of the adsorption in the saturation level was found to be P123 approximately P103 > P105 >> F108, meaning that Pluronics with higher hydrophobicity can adsorb preferentially to the silica surface. Indeed, this order was parallel to the order of the adsolubilization amount of 2-naphthol. In the case of co-addition of the Pluronics and 2-naphthol, the adsolubilization amount increased gradually at lower surfactant concentration regions, reached a maximum, and then decreased with increasing concentration of the Pluronics. The maximum amount appeared at critical polymolecular micelle concentration of each Pluronics. On the other hand, the final decrement was not observed when 2-naphthol was added after replacement of the Pluronics supernatant by the Pluronics free solution. These results suggest that adsolubilization behavior is influenced by the existence of the polymolecular micellar aggregates in the solution phase. PMID- 16376928 TI - Histopathology of idiopathic lesions in the eyes of Homarus americanus from Long Island Sound. AB - In 1999, American lobsters, Homarus americanus, from western Long Island Sound (WLIS) experienced a significant mortality. In 2001 and 2004, the eyes and eyestalks of lobsters from WLIS and central LIS were examined for histopathological changes. Idiopathic lesions were identified in the ommatidia and optic nerve fibers proximal to the ommatidia in 29 (56%) of the lobsters from LIS. Lesions were categorized as either moderate or severe. Moderate lesions had altered rhabdoms, clumped pigment, and altered optic nerve fibers. Severe lesions were marked by absent rhabdoms, clumped pigment in both the ommatidial region and in the optic nerve region; and optic nerve fibers that had been completely destroyed and were replaced by vascular tissue. Idiopathic lesions occurred primarily in the central and ventral regions of the eye, and with much less frequency in the dorsal region. In addition, damage to the dorsal area tended to occur only when the severity of lesions was high, indicating a spatially progressive pattern to the lesion development. The lesions occurred in both western and central Long Island Sound, with no significant differences in severity between locations. The prevalence of lesions did not vary between years, but in 2004, several eyes had less severe pathology than those from 2001. These data indicate that the etiological agent is present throughout a large portion of the Sound, and that lobsters are probably continually exposed to it. PMID- 16376929 TI - Phagocytosis by Lymnaea stagnalis haemocytes: a potential role for phosphatidylinositol 3-kinase but not protein kinase A. AB - The molecular events that regulate phagocytosis, an important innate immune response, in invertebrate defence cells (haemocytes) are poorly understood. Lymnaea stagnalis haemocytes were used as a model to elucidate the role of cell signalling pathways in phagocytosis by molluscan defence cells. The phosphatidylinositol 3-kinase (PI3-K) inhibitor, LY294002, significantly impaired haemocyte phagocytic activity in a dose-responsive manner with 10 microM LY294002 reducing internalization of fluorescent-conjugated Escherichia coli by 62% (P < or = 0.001). In contrast, the protein kinase A (PKA) inhibitor KT5720 was without effect. Therefore, PI3-K, but not PKA, appears to control phagocytosis by haemocytes in these gastropod molluscs. PMID- 16376930 TI - A scientific note on the detection of honeybee viruses using real-time PCR (TaqMan) in Varroa mites collected from a Thai honeybee (Apis mellifera) apiary. AB - Bee parasitic mite syndrome is a disease complex of colonies simultaneously infested with Varroa destructor mites and infected with viruses and accompanied by high mortality. By using real-time PCR (TaqMan), five out of seven bee viruses were detected in mite samples (V. destructor) collected from Thailand. Moreover, the results of this study provide an evidence for the co-existence of several bee viruses in a single mite. This is also the first report of bee viruses in mites from Thailand. PMID- 16376931 TI - Molecular analysis of an occlusion body protein from Culex nigripalpus nucleopolyhedrovirus (CuniNPV). AB - CUN085 is an occlusion body (OB) protein from the nucleopolyhedrovirus of Culex nigripalpus (CuniNPV). SDS-PAGE analysis indicated that the CuniNPV OB protein is about 3 times the size (approximately 90 kDa) of characterized nucleopolyhedrovirus (NPVs) and granulovirus OB proteins. Rapid amplification of cDNA ends (RACE), RNase protection assay, real-time PCR, and protein sequencing were used to characterize CUN085 from CuniNPV. RACE data revealed that the transcriptional start and termination sites for the CUN085 gene yielded a polypeptide comprised of 822 amino acids indicating that translation initiates within a larger 882 amino acid open reading frame that was originally predicted from the CuniNPV genome sequence. Transcription of CUN085 started at a consensus baculovirus late transcription start site TAAG at nucleotide position 75433 of the CuniNPV genome sequence. RNase protection assays and quantitative real-time PCR show that the CUN085 transcript is first detected in mosquito larvae at approximately 6 h after infection with CuniNPV and its prevalence increased progressively over the subsequent 18 h. PMID- 16376932 TI - Molecular and biological characterization of a Cypovirus from the mosquito Culex restuans. AB - A cypovirus from the mosquito Culex restuans (named CrCPV) was isolated and its biology, morphology, and molecular characteristics were investigated. CrCPV is characterized by small (0.1-1.0 microm), irregularly shaped inclusion bodies that are multiply embedded. Laboratory studies demonstrated that divalent cations influenced transmission of CrCPV to Culex quinquefasciatus larvae; magnesium enhanced CrCPV transmission by approximately 30% while calcium inhibited transmission. CrCPV is the second cypovirus from a mosquito that has been confirmed by using molecular analysis. CrCPV has a genome composed of 10 dsRNA segments with an electropherotype similar to the recently discovered UsCPV-17 from the mosquito Uranotaenia sapphirina, but distinct from the lepidopteran cypoviruses BmCPV-1 (Bombyx mori) and TnCPV-15 (Trichoplusia ni). Nucleotide and deduced amino acid sequence analysis of CrCPV segment 10 (polyhedrin) suggests that CrCPV is closely related (83% nucleotide sequence identity and 87% amino acid sequence identity) to the newly characterized UsCPV-17 but is unrelated to the 16 remaining CPV species from lepidopteran hosts. A comparison of the terminal segment regions of CrCPV and UsCPV-17, an additional method for differentiating various Cypovirus species, revealed a high level of conservation. Therefore, we propose that CrCPV is a member of the Cypovirus-17 group and designate this species as CrCPV-17. PMID- 16376934 TI - Purification, properties and extended solution structure of the complex formed between human immunoglobulin A1 and human serum albumin by scattering and ultracentrifugation. AB - Immunoglobulin A (IgA) is unique amongst antibodies in being able to form polymeric structures that may possess important functions in the pathology of specific diseases. IgA also forms complexes with other plasma proteins, the IgA1 human serum albumin (HSA) complex (IgA1-HSA) being typical. We have purified this complex using a novel two-step purification based on thiophilic chromatography and gel filtration, and characterised this. HSA is linked covalently to the tailpiece of IgA1 by a disulphide bond between Cys471 in IgA1 and Cys34 in HSA. IgA1-HSA binds to IgA receptors on neutrophils and monocytes, and elicits a respiratory burst that is comparable in magnitude to that of monomeric IgA1. The solution arrangement of IgA1-HSA was identified by X-ray scattering and ultracentrifugation. The radius of gyration R(G) of 7.5(+/-0.3) nm showed that IgA1-HSA is more extended in solution than IgA1 (R(G) of 6.1-6.2 nm). Its distance distribution function P(r) showed two peaks that indicated a well separated solution structure similar to that for IgA1, and a maximum dimension of 25 nm, which is greater than that of 21 nm for IgA1. Sedimentation equilibrium showed that the IgA1:HSA stoichiometry is 1:1. Sedimentation velocity resulted in a sedimentation coefficient of 6.4S and a frictional ratio of 1.87, which is greater than that of 1.56 for IgA1. The constrained modelling of the IgA1-HSA structure using known structures for IgA1 and HSA generated 2432 conformationally randomised models of which 52 gave good scattering fits. The HSA structure was located at the base of the Fc fragment in IgA1 in an extended arrangement. Such a structure accounts for the functional activity of IgA1-HSA, and supports our previous modelling analysis of the IgA1 solution structure. The IgA1-HSA complex may suggest the potential for creating a new class of targeted therapeutic reagents based on the coupling of IgA1 to carrier proteins. PMID- 16376933 TI - Multiple U2AF65 binding sites within SF3b155: thermodynamic and spectroscopic characterization of protein-protein interactions among pre-mRNA splicing factors. AB - Essential, protein-protein complexes between the large subunit of the U2 small nuclear RNA auxiliary factor (U2AF65) with the splicing factor 1 (SF1) or the spliceosomal component SF3b155 are exchanged during a critical, ATP-dependent step of pre-mRNA splicing. Both SF1 and the N-terminal domain of SF3b155 interact with a U2AF homology motif (UHM) of U2AF65. SF3b155 contains seven tryptophan containing sites with sequence similarity to the previously characterized U2AF65 binding domain of SF1. We show that the SF3b155 domain lacks detectable secondary structure using circular dichroism spectroscopy, and demonstrate that five of the tryptophan-containing SF3b155 sites are recognized by the U2AF65-UHM using intrinsic tryptophan fluorescence experiments with SF3b155 variants. When compared with SF1, similar spectral shifts and sequence requirements indicate that U2AF65 interactions with each of the SF3b155 sites are similar to the minimal SF1 site. However, thermodynamic comparison of SF1 or SF3b155 proteins with minimal peptides demonstrates that formation the SF1/U2AF65 complex is likely to affect regions of SF1 beyond the previously identified, linear interaction site, in a remarkably distinct manner from the local U2AF65 binding mode of SF3b155. Furthermore, the complex of the SF1/U2AF65 interacting domains is stabilized by 3.3 kcal mol-1 relative to the complex of the SF3b155/U2AF65 interacting domains, consistent with the need for ATP hydrolysis to drive exchange of these partners during pre-mRNA splicing. We propose that the multiple U2AF65 binding sites within SF3b155 regulate conformational rearrangements during spliceosome assembly. Comparison of the SF3b155 sites defines an (R/K)nXRW(DE) consensus sequence for predicting U2AF65-UHM ligands from genomic sequences, where parentheses denote residues that contribute to, but are not required for binding. PMID- 16376935 TI - Diversification of catalytic activities and ligand interactions in the protein fold shared by the sugar isomerases, eIF2B, DeoR transcription factors, acyl-CoA transferases and methenyltetrahydrofolate synthetase. AB - Evolution of diverse catalytic and ligand-binding activities in a given protein fold is a widely observed phenomenon in the protein-domain universe. However, the details of this evolutionary process, general principles, if any, and implications for origins of particular catalytic mechanisms are poorly understood in many common protein folds. Taking advantage of the wealth of currently available protein structure and sequence data, we explore these issues in the context of a large assemblage of biochemically diverse protein domains sharing a common origin, namely the sugar isomerases, translation factor eIF2B, ligand binding domains of the DeoR-family transcription factors, acetyl-CoA transferases and methenyltetrahydrofolate synthetase. We show that in at least three independent instances, including the sugar-binding domains of the DeoR family transcription factors, this domain has been used as small molecule sensor coupled to helix-turn-helix DNA-binding domains. In at least two of these instances the domain functions as a non-catalytic sensor of ligands. We provide evidence that the ancestral version of this fold was a distinct version of the Rosmann-like folds, which probably possessed two distinct ligand-binding areas that were differentially utilized in different descendents. Analyzing the sequences and structures of proteins in this fold we show that there are two principal factors related to the origin of catalytic diversity in this fold. Firstly, specific inserts and extension added to the core domain on multiple occasions in evolution have affected the access to the active site regions, and thereby allowed for different substrates and allosteric regulators. The second major factor appears to be the emergence of considerable diversity of family-specific residues with important biochemical roles. Interestingly, proteins of this fold, which catalyze similar reactions on similar substrates, might possess very distinctive sets of active residues required for substrate binding catalysis. In particular, different sugar isomerases or acyl transferases in this fold might show distinct constellations of active site residues. These findings suggest that whereas ligand-binding, and even generic catalytic ability emerged early in the evolution of the fold, the specific catalytic mechanisms appear to have independently emerged on multiple occasions in the generic precursors of this fold. PMID- 16376936 TI - Cis and trans-acting effects on a mutational hotspot involving a replication template switch. AB - A natural mutational hotspot in the thyA gene of Escherichia coli accounts for over half of the mutations that inactivate this gene, which can be selected by resistance to the antibiotic trimethoprim. This T to A transversion, at base 131 of the coding sequence, occurs within a 17 bp quasi-palindromic sequence. To clarify the mechanism of mutagenesis, we examine here cis and trans-acting factors affecting thyA131 mutational hotspot activity at its natural location on the E.coli chromosome. Confirming a template-switch mechanism for mutagenesis, an alteration that strengthens base-pairing between the inverted repeat DNA sequences surrounding the hotspot stimulated mutagenesis and, conversely, mutations that weakened pairing reduced hotspot activity. In addition, consistent with the idea that the hotspot mutation is templated from DNA synthesis from mispaired strands of the inverted repeats, co-mutation of multiple sites within the quasipalindrome was observed as predicted from the DNA sequence of the corresponding repeat. Surprisingly, inversion of the thyA operon on the chromosome did not abolish thyA131 hotspot mutagenesis, indicating that mutagenesis at this site occurs during both leading and lagging-strand synthesis. Loss of the SOS-induced DNA polymerases PolII, PolIV, and PolV, caused a marked increase in the hotspot mutation rate, indicating a heretofore unknown and redundant antimutagenic effect of these repair polymerases. Hotspot mutagenesis did not require the PriA replication restart factor and hence must not require fork reassembly after the template-switch reaction. Deficiency in the two major 3' single-strand DNA exonucleases, ExoI and ExoVII, stimulated hotspot mutagenesis 30-fold and extended the mutagenic tract, indicating that these exonucleases normally abort a large fraction of premutagenic events. The high frequency of quasipalindrome-associated mutations suggests that template switching occurs readily during chromosomal replication. PMID- 16376937 TI - Structural bases of feed-back control of arginine biosynthesis, revealed by the structures of two hexameric N-acetylglutamate kinases, from Thermotoga maritima and Pseudomonas aeruginosa. AB - N-Acetylglutamate kinase (NAGK) catalyses the second step in the route of arginine biosynthesis. In many organisms this enzyme is inhibited by the final product of the route, arginine, and thus plays a central regulatory role. In addition, in photosynthetic organisms NAGK is the target of the nitrogen signalling protein PII. The 3-D structure of homodimeric, arginine-insensitive, Escherichia coli NAGK, clarified substrate binding and catalysis but shed no light on arginine inhibition of NAGK. We now shed light on arginine inhibition by determining the crystal structures, at 2.75 A and 2.95 A resolution, of arginine complexed Thermotoga maritima and arginine-free Pseudomonas aeruginosa NAGKs, respectively. Both enzymes are highly similar ring-like hexamers having a central orifice of approximately 30 A diameter. They are formed by linking three E.coli NAGK-like homodimers through the interlacing of an N-terminal mobile kinked alpha helix, which is absent from E.coli NAGK. Arginine is bound in each subunit of T.maritima NAGK, flanking the interdimeric junction, in a site formed between the N helix and the C lobe of the subunit. This site is also present, in variable conformations, in P.aeruginosa NAGK, but is missing from E.coli NAGK. Arginine, by gluing the C lobe of each subunit to the inter-dimeric junction, may stabilize an enlarged active centre conformation, hampering catalysis. Acetylglutamate counters arginine inhibition by promoting active centre closure. The hexameric architecture justifies the observed sigmoidal arginine inhibition kinetics with a high Hill coefficient (N approximately 4), and appears essential for arginine inhibition and for NAGK-PII complex formation, since this complex may involve binding of NAGK and PII with their 3-fold axes aligned. The NAGK structures allow identification of diagnostic sequence signatures for arginine inhibition. These signatures are found also in the homologous arginine-inhibited enzyme NAG synthase. The findings on NAGK shed light on the structure, function and arginine inhibition of this synthase, for which a hexameric model is constructed. PMID- 16376938 TI - Interaction of gp16 with pRNA and DNA for genome packaging by the motor of bacterial virus phi29. AB - One striking feature in the assembly of linear double-stranded (ds) DNA viruses is that their genome is translocated into a preformed protein coat via a motor involving two non-structural components with certain characteristics of ATPase. In bacterial virus phi29, these two components include the protein gp16 and a packaging RNA (pRNA). The structure and function of other phi29 motor components have been well elucidated; however, studies on the role of gp16 have been seriously hampered by its hydrophobicity and self-aggregation. Such problems caused by insolubility also occur in the study of other viral DNA-packaging motors. Contradictory data have been published regarding the role and stoichiometry of gp16, which has been reported to bind every motor component, including pRNA, DNA, gp3, DNA-gp3, connector, pRNA-free procapsid, and procapsid/pRNA complex. Such conflicting data from a binding assay could be due to the self-aggregation of gp16. Our recent advance to produce soluble and highly active gp16 has enabled further studies on gp16. It was demonstrated in this report that gp16 bound to DNA non-specifically. gp16 bound to the pRNA-containing procapsid much more strongly than to the pRNA-free procapsid. The domain of pRNA for gp16 interaction was the 5'/3' paired helical region. The C18C19A20 bulge that is essential for DNA packaging was found to be dispensable for gp16 binding. This result confirms the published model that pRNA binds to the procapsid with its central domain and extends its 5'/3' DNA-packaging domain for gp16 binding. It suggests that gp16 serves as a linkage between pRNA and DNA, and as an essential DNA-contacting component during DNA translocation. The data also imply that, with the exception of the C18C19A20 bulge, the main role of the 5'/3' helical double-stranded region of pRNA is not for procapsid binding but for binding to gp16. PMID- 16376939 TI - Retention of misfolded mutant transthyretin by the chaperone BiP/GRP78 mitigates amyloidogenesis. AB - Carriers of the D18G transthyretin (TTR) mutation display an unusual central nervous system (CNS) phenotype with late onset of disease. D18G TTR is monomeric and highly prone to misfold and aggregate even at physiological conditions. Extremely low levels of mutant protein circulate both in human serum and cerebrospinal fluid, indicating impaired secretion of D18G TTR. Recent data show efficient selective ER-associated degradation (ERAD) of D18G TTR. One essential component of the ER-assisted folding machinery is the molecular chaperone BiP. Co expression of BiP and D18G TTR, or BiP and wild-type (wt) TTR, or mutants A25T TTR and L55P TTR in Escherichia coli showed that only D18G TTR was significantly captured by BiP. Negligible capture of wt TTR and L55P TTR was seen and a sixfold smaller amount of A25T TTR bound to BiP compared to D18G TTR. These data correlate very well with thermodynamic and kinetic stability of the TTR variants, indicating that folding efficiency is inversely correlated to BiP capture. The complexes between BiP and D18G TTR were stable and could be isolated through affinity chromatography. Analytical ultracentrifugation and size-exclusion chromatography revealed that D18G TTR and BiP formed a mixture of 1:1 complexes and large soluble oligomers. The stoichiometry of captured D18G TTR versus BiP increased with increasing size of the oligomers. This indicates that BiP either worked as a molecular shepherd collecting the aggregation-prone mutant into stable oligomers or that BiP could bind to oligomers formed from misfolded mutant protein. Sequence analysis of bound TTR peptides to BiP revealed a bound sequence corresponding to residues 88-103 of TTR, comprising beta-strand F in the folded TTR monomer constituting part of the hydrogen bonding tetramer interface in native TTR. The F-strand has also been suggested as a possible elongation region of amyloid fibrils, implicating how substoichiomeric amounts of BiP could sequester prefibrillar amyloidogenic oligomers through binding to this part of TTR. BiP binding to D18G TTR was abolished by addition of ATP. The released D18G TTR completely misfolded into amyloid aggregates as shown by ThT fluorescence and Congo red binding. PMID- 16376940 TI - Regulation of Fas-mediated apoptosis in neutrophils after surgery-induced acute inflammation. AB - BACKGROUND: Neutrophils undergo rapid Fas-mediated apoptosis during in vitro culture. The purpose of this study was to investigate the effects of surgical stress upon the Fas-mediated apoptotic response in circulating neutrophils. MATERIALS AND METHODS: Blood samples were drawn from eight patients with a mandibular prognathism, and who had undergone a bilateral sagittal split ramus osteotomy, at 2 days before, and at 1 and 5 days after surgery. The circulating neutrophils in each blood sample were then evaluated for their susceptibility to Fas-mediated apoptosis in either the presence or the absence of autogenous plasma. RESULTS: Fas-induced apoptosis in the neutrophils of these surgically treated patients was found to be slightly accelerated at 1 day postoperatively in the presence of FBS, compared with 2 days preoperatively and 5 days postoperatively. However, we obtained different results for these experiments in the presence of autogenous plasma. The Fas-induced apoptotic response levels in the neutrophils at day 1 postsurgery following exposure to autogenous plasma were significantly suppressed compared with the levels at both 2 days preoperatively and 5 days postoperatively. The Fas expression levels on the cell surface of the neutrophils were not altered, but the levels of soluble Fas (sFas) in the plasma were reduced to almost inverse levels during the postoperative periods. The levels of granulocyte-macrophage colony-stimulating factor, interleukin-6, and interleukin-8 levels in the plasma were also markedly raised in the plasma from each of these patients at 1 day postoperatively. However, the anti-apoptotic effects of the plasma on the Fas-mediated neutrophil apoptosis were not influenced by the addition of their neutralizing antibodies for these cytokines. The suppressive effects of postoperative plasma on Fas-mediated neutrophil apoptosis were blocked by the phosphatidylinositol 3-kinase (PI 3-K) inhibitors, LY294002, and wortmannin. Additionally, these effects were also abrogated by the extracellular signal-regulated kinase (ERK) inhibitor, PD98059, but not by the p38 mitogen-activated protein kinase inhibitor, SB203580. CONCLUSIONS: The increase in sFas levels in the plasma of patients with acute inflammation may lead to the inhibition of Fas-mediated neutrophil apoptosis. Moreover, the activation of the PI 3-K and ERK signaling-dependent pathways may, in part, also contribute to the down-regulation of the Fas-mediated apoptotic response in neutrophils. PMID- 16376941 TI - An ergonomic comparison of robotic and laparoscopic technique: the influence of surgeon experience and task complexity. AB - BACKGROUND: This study compares the mental and physical workload of laparoscopic and robotic technique while performing simulated surgical tasks in a laboratory setting. MATERIALS AND METHODS: Ten volunteer surgeons performed two tasks in a laparoscopic trainer using laparoscopic (LAP) and robotic (ROB) techniques. Outcome measures included: Task time, task-error, vertical/horizontal arm displacement, percent maximum electromyographic signal from the thenar, forearm flexor, and deltoid muscle compartments, skin conductance, and perceived difficulty and discomfort levels. A two-way repeated-measures ANOVA compared surgical technique and laparoscopic experience level (E = expert, N = novice). RESULTS: For the simple task, ROB technique was slower and had higher errors, and the surgeon's arm was more elevated. For the complex task, ROB electromyographic signal was lower. Stress was lower in both tasks for ROB, but the decrease was not statistically significant. CONCLUSIONS: Robotic technique appears slower and less precise than laparoscopic technique for simple tasks, but equally fast and possibly less stressful for complex tasks. Previous laparoscopic experience has a complex influence on the physical and mental adaptation to robotic surgery. PMID- 16376942 TI - Uncommon V599E BRAF mutations in Japanese patients with lung cancer. AB - BACKGROUND: Recently, somatic mutations of the epidermal growth factor receptor (EGFR) gene and Braf gene were found in patients with lung cancer. These mutations might be correlated with a clinical response to molecular target therapy. Although a few Caucasian lung cancer patients harbored BRAF mutations, there have been no reports about the BRAF mutation in Japanese patients with lung cancer. MATERIALS AND METHODS: We investigated BRAF mutations in Japanese lung cancer patients. The study included 129 surgically removed lung cancer cases from Nagoya City University Hospital. Braf, EGFR, and erbB2 mutations also were analyzed by reverse transcript polymerase chain reaction (RT-PCR) and direct sequencing. RESULTS: In exon 15, one BRAF mutation (1796 thymine to adenine; V599E) was found in nonsmoking woman with well-differentiated adenocarcinoma. We detected the 43 EGFR mutations, including 19 at exon 19 and 20 at exon 21 from the 129 patients analyzed (33.3%). We also detected one erbB2 mutation from 76 patients analyzed. All these mutations existed exclusively. CONCLUSIONS: V599E BRAF mutation was uncommon in Japanese lung cancer. All three genes mutations were predominantly found in female nonsmoking subjects with adenocarcinomas. However, completely exclusive mutation status would help us to choose custom-made molecular target therapy for the lung cancer. PMID- 16376943 TI - Efficacy of transient treatment with FK506 in the early phase on cyclophosphamide induced bone marrow chimerism and transplant tolerance across MHC barriers. AB - BACKGROUND: The use of mixed allogeneic bone marrow chimerism to induce donor specific transplantation tolerance has been extensively demonstrated. In the present study, we assessed the effect of combined use of a short course of FK506 and a single-dose cyclophosphamide (CYP) on the induction of tolerance and development of GVHD after allogeneic BMT. MATERIALS AND METHODS: Lewis rat (RT1(l)) recipients received BMT from Brown Norway (RT1(n)) donors on the next day after injection of CYP at a dose of 200 mg/kg. The recipients were further treated with no FK506 (n = 8), 0.3 mg/kg/day FK506 on days 10-16 (n = 6), or the same dose of FK506 on days 0-6 (n = 6). In a subgroup of animals, heterotopic heart transplantation was performed to investigate transplantation tolerance. RESULTS: Six of eight recipient rats that did not receive FK506 died of severe GVHD, while high levels of chimerism were induced. Recipients of FK506 in the later phase developed mild transient GVHD around 2 to 3 weeks after BMT and recovered thereafter; however, the level of chimerism was significantly decreased (2.8 +/- 2.3% on day 100). Treatment with FK506 in the early phase completely prevented the development of GVHD and induced stable allogeneic chimerism in the long-term (13.8 +/- 8.3% on day 100). These recipients with stable chimerism accepted subsequent BN heart allografts indefinitely (>200 days x 5), while rejecting third-party (BUF) heart allografts by day 12. CONCLUSIONS: Early transient FK506 promotes the induction of stable bone marrow chimerism without GVHD after BMT with CYP pretreatment. The timing of treatment with FK506 is critical with a view to preventing GVHD and inducing stable long-lasting chimerism. PMID- 16376944 TI - Upregulation of vascular growth factors in multiple sclerosis: correlation with MRI findings. AB - Vascular permeability changes precede the development of demyelinating lesions in multiple sclerosis (MS), and vessel wall thickening and capillary proliferation are frequently seen in autopsied MS lesions. Although vascular growth factors are critical for inducing such vascular changes, their involvement in MS has not been extensively studied. Thus, we examined the involvement of various vascular growth factors in MS according to their clinical phase and subtype. We measured serum levels of vascular endothelial growth factor (VEGF), acidic and basic fibroblast growth factors (FGF) and platelet-derived growth factors (PDGFs)-AA, -AB and -BB in 50 patients with MS (27 opticospinal MS and 23 conventional MS patients) and 33 healthy controls using sandwich enzyme immunoassays. Correlations between growth factor changes and brain and spinal cord MRI findings were then analyzed. Serum VEGF concentrations were significantly higher in MS patients in relapse than in controls (p = 0.0495) and in MS patients in remission (p = 0.0003), irrespective of clinical subtype. Basic FGF was significantly increased in conventional MS patients, but not opticospinal MS patients compared with controls (p = 0.0291), irrespective of clinical phase. VEGF at relapse showed a significant positive correlation with the length of spinal cord lesions on MRI (r = 0.506, p = 0.0319). The results suggest that an increase in serum VEGF concentration might be involved in MS relapse and the formation of longitudinally extensive spinal cord lesions. PMID- 16376946 TI - An alternative formulation for a delayed logistic equation. AB - We derive an alternative expression for a delayed logistic equation, assuming that the rate of change of the population depends on three components: growth, death, and intraspecific competition, with the delay in the growth component. In our formulation, we incorporate the delay in the growth term in a manner consistent with the rate of instantaneous decline in the population given by the model. We provide a complete global analysis, showing that, unlike the dynamics of the classical logistic delay differential equation (DDE) model, no sustained oscillations are possible. Just as for the classical logistic ordinary differential equation (ODE) growth model, all solutions approach a globally asymptotically stable equilibrium. However, unlike both the logistic ODE and DDE growth models, the value of this equilibrium depends on all of the parameters, including the delay, and there is a threshold that determines whether the population survives or dies out. In particular, if the delay is too long, the population dies out. When the population survives, i.e., the attracting equilibrium has a positive value, we explore how this value depends on the parameters. When this value is positive, solutions of our DDE model seem to be well approximated by solutions of the logistic ODE growth model with this carrying capacity and an appropriate choice for the intrinsic growth rate that is independent of the initial conditions. PMID- 16376947 TI - The astroglial-derived S100beta protein stimulates the expression of nitric oxide synthase in rodent macrophages through p38 MAP kinase activation. AB - S100beta is an astroglial-derived Ca2+ -binding protein having neurotrophic role on neurons and glial cells. An aberrant S100beta production has been observed in neurodegenerative disease, as Alzheimer's disease and Down syndrome. S100beta is responsible to start up a gliotic reaction by the release of pro-inflammatory mediators, including nitric oxide (NO) and cytokines from microglia and astrocytes, which are, in turn, deleterious for neurons. Interestingly, pro inflammatory effect of S100beta seems not be restricted into the brain. Macrophages play a pivotal role in inflammatory diseases, occurring both in the brain and in the periphery. In this study, we tested the hypothesis that S100beta may affect macrophage functions, amplifying thus the inflammatory process. Our results demonstrate that S100beta stimulates both NO production and iNOS protein transcription and expression in J774 and rat peritoneal macrophages. NO production was concentration and time-dependently inhibited by two iNOS inhibitors, L-NAME and SMT. We also demonstrated that S100beta induced oxidative stress by increasing H2O2 production and lipid peroxidation of cell membrane in both macrophage types. The pro-oxidant potential of S100beta activates p38 MAP kinase (MAPK), which has been described to directly activate NF-kappaB. In our study, SB203580, a p38 MAPK inhibitor, and two NF-kappaB inhibitors, TLCK and BAY 11-7082, decreased both NO production and iNOS protein transcription and expression in S100beta-stimulated J774 and peritoneal rat macrophages. Moreover, additional studies demonstrated that S100beta affected also TNF-alpha protein expression in J774 macrophages. In conclusion, our results highlight the potential role of S100beta during an inflammatory scenario identifying macrophages as a novel S100beta-responsive cell-type. PMID- 16376949 TI - Improvement of pollutant drift forecast system applied to the Prestige oil spills in Galicia Coast (NW of Spain): development of an operational system. AB - An integrated system named METEOMOHID, developed by MeteoGalicia in the first stage of the Prestige accident in November 2002 was used successfully in an operational form to support decision making and assist in recovering tasks. Afterwards, METEOMOHID has been enhanced with the aim of developing an operational oceanography system to be used in the NW of the Iberian Peninsula. The METEOMOHID system includes local area hydrodynamic coastal ocean modelling (MOHID), real time atmospheric forcing from a local meteorological model (ARPS). Using the available data from the Prestige crisis, a set of simulations were designed in order to reproduce the oil spill drift. The implementation of a detailed vertical resolution in the model has allowed obtaining a detailed surface dynamic, improving our knowledge of the behaviour of tarballs into the water column. Thus, the wind-driven Eckman drift, the direct dragging of the wind were detached, and the possible existence of subsurface oil was assessed. In addition, the present work evaluates the effects of introducing climatologic large scale currents in the METEOMOHID system. PMID- 16376948 TI - Selective modulation of L-type calcium current by magnesium lithospermate B in guinea-pig ventricular myocytes. AB - Magnesium lithospermate B (MLB) is the main water-soluble principle of Salviae Miltiorrhizae Radix (also called as 'Danshen' in the traditional Chinese medicine) for the treatment of cardiovascular diseases. MLB was found to possess a variety of pharmacological actions. However, it is unclear whether and how MLB affects the cardiac ion channels. In the present study, the effects of MLB on the voltage-activated ionic currents were investigated in single ventricular myocytes of adult guinea pigs. MLB reversibly inhibited L-type Ca(2+) current (I(Ca,L)). The inhibition was use-dependent and voltage-dependent (the IC(50) value of MLB was 30 microM and 393 microM, respectively, at the holding potential of -50 mV and -100 mV). In the presence of 100 microM MLB, both the activation and steady state inactivation curves of I(Ca,L) were markedly shifted to hyperpolarizing membrane potentials, whereas the time course of recovery of I(Ca,L) from inactivation was not altered. MLB up to 300 microM had no significant effect on the fast-inactivating Na(+) current (I(Na)), delayed rectifier K(+) current (I(K)) and inward rectifier K(+) current (I(K1)). The results suggest that the voltage-dependent Ca(2+) antagonistic effect of MLB work in concert with its antioxidant action for attenuating heart ischemic injury. PMID- 16376950 TI - Mathematical modeling of cancer radiovirotherapy. AB - Cancer virotherapy represents a dynamical system that requires mathematical modeling for complete understanding of the outcomes. The combination of virotherapy with radiation (radiovirotherapy) has been recently shown to successfully eliminate tumors when virotherapy alone failed. However, it introduces a new level of complexity. We have developed a mathematical model, based on population dynamics, that captures the essential elements of radiovirotherapy. The existence of corresponding equilibrium points related to complete cure, partial cure, and therapy failure is proved and discussed. The parameters of the model were estimated by fitting to experimental data. By using simulations we analyzed the influence of parameters that describe the interaction between virus and tumor cell on the outcome of the therapy. Furthermore, we evaluated relevant therapeutic scenarios for radiovirotherapy, and offered elements for optimization. PMID- 16376951 TI - Albumin leak across human pulmonary microvascular vs. umbilical vein endothelial cells under septic conditions. AB - Human pulmonary microvascular endothelial cell (HPMVEC) injury is central to the pathophysiology of human lung injury. However, septic HPMVEC barrier dysfunction and the contribution of neutrophils have not been directly addressed in vitro. Instead, human EC responses are often extrapolated from studies of human umbilical vein EC (HUVEC). We hypothesized that HUVEC was not a good model for investigating HPMVEC barrier function under septic conditions. HPMVEC was isolated from lung tissue resected from lung cancer patients using magnetic bead bound anti-PECAM-1 antibody. In confluent monolayers in 3-mum cell-culture inserts, we assessed trans-EC Evans-Blue (EB)-conjugated albumin leak under basal, unstimulated conditions and following stimulation with either lipopolysaccharide or a mixture of equal concentrations of TNF-alpha, IL-1beta and IFN-gamma (cytomix). Basal EB-albumin leak was significantly lower across HPMVEC than HUVEC (0.64 +/- 0.06% vs. 1.13 +/- 0.10%, respectively, P < 0.001). Lipopolysaccharide and cytomix increased leak across both HPMVEC and HUVEC in a dose-dependent manner, with a similar increase relative to basal leak in both cell types. The presence of neutrophils markedly and dose-dependently enhanced cytomix-induced EB-albumin leak across HPMVEC (P < 0.01), but had no effect on EB albumin leak across HUVEC. Both cytomix and lipopolysaccharide-induced albumin leak was not associated with a loss of cell viability. In conclusion, HPMVEC barrier dysfunction under septic conditions is dramatically enhanced by neutrophil presence, and HUVEC is not a suitable model for studying HPMVEC septic barrier responses. The direct study of HPMVEC septic responses will lead to a better understanding of human lung injury. PMID- 16376952 TI - Suppression of post-hypoxic-ischemic EEG transients with dizocilpine is associated with partial striatal protection in the preterm fetal sheep. AB - In vitro studies suggest that glutamate receptor activation is important in the genesis of post-hypoxic preterm brain injury, but there are limited data on post hypoxic N-methyl-D-aspartate (NMDA) receptor activation. We therefore examined an infusion of the specific, non-competitive NMDA receptor antagonist dizocilpine (2 mg kg(-1) bolus plus 0.07 mg kg(-1) h(-1) i.v.) from 15 min to 4 h after severe hypoxia-ischemia induced by umbilical cord occlusion for 25 min in fetal sheep at 70% of gestation. Dizocilpine suppressed evolving epileptiform transient activity in the first 6 h after reperfusion (2.3 +/- 0.9 versus 9.3 +/- 2.3 maximal counts min(-1), P < 0.05) and mean EEG intensity up to 11 h after occlusion (P < 0.05). Fetal extradural temperature transiently increased during the dizocilpine infusion (40.1 +/- 0.2 versus 39.3 +/- 0.1 degrees C, P < 0.05). After 3 days recovery, treatment was associated with a significant reduction in neuronal loss in the striatum (31 +/- 7 versus 58 +/- 2%, P < 0.05), expression of cleaved caspase-3 (111+/-7 versus 159 +/- 10 counts area(-1), P < 0.05) and numbers of activated microglia (57 +/- 9 versus 92 +/- 16 counts area(-1), P < 0.05); there was no significant effect in other regions or on loss of immature O4-positive oligodendrocytes. In conclusion, abnormal NMDA receptor activation in the first few hours of recovery from hypoxia-ischemia seems to contribute to post-hypoxic striatal damage in the very immature brain. PMID- 16376953 TI - D2 dopamine receptor-induced sensitization of adenylyl cyclase type 1 is G alpha(s) independent. AB - Acute activation of D2 dopamine receptors inhibits adenylyl cyclase (EC 4.6.1.1), whereas persistent activation of these inhibitory receptors results in a compensatory increase in cyclic AMP accumulation. This sensitization of adenylyl cyclase is thought to involve enhanced Galpha(s)-adenylyl cyclase interactions; however, the absolute requirement of Galpha(s) has not been determined. The present study used a Galpha(s)-deficient cell line to examine directly the role of Galpha(s) in D2 dopamine receptor-induced sensitization of recombinant adenylyl cyclase type 1 (AC1) and 5 (AC5). In acute experiments, quinpirole activation of the D2 dopamine receptor inhibited AC1 and AC5 activity, indicating that the acute regulatory properties of AC1 and AC5 were retained in the absence of Galpha(s). Subsequent experiments revealed that short-term (2 h) activation of the D2 dopamine receptor resulted in significantly enhanced forskolin-stimulated AC1 activity in the absence of Galpha(s), whereas sensitization of forskolin stimulated AC5 activity appeared to require Galpha(s). The Galpha(s)-independent sensitization of AC1 was explored further using AC1-selective activation protocols (A23187 and CCE) following short- and long-term agonist treatment. These studies revealed that persistent activation of D2 dopamine receptors sensitized AC1 activity to Ca2+ stimulation in cells devoid of endogenous Galpha(s) and demonstrate directly that sensitization of AC1 is Galpha(s) independent. PMID- 16376954 TI - Dissociating cognitive deficits involved in voluntary eye movement dysfunctions in Parkinson's disease patients. AB - In an attempt to distinguish and define the altered cognitive processes associated with Parkinson's disease (PD), we examine and try to dissociate the components of an effective voluntary saccade: (1) the planning and execution of a voluntary saccade; (2) the suppression of reflexive eye movements; and (3) the working memory processes required. We tested 14 PD patients (off their medications) and 11 control subjects on antisaccade (AS), delayed antisaccade (DAS), and remembered antisaccade (RAS) paradigms. The three tasks required identical responses, each task only differing in a single manipulation for direct comparison--a delay period was added in the DAS, and the target was removed during the delay period of the RAS--allowing us to study the specific cognitive processes involved in the execution of a voluntary saccade. Voluntary saccade response times were longer in the PD group compared to controls on all three tasks, suggesting difficulties in voluntary saccade execution. Furthermore, PD patients showed difficulty suppressing reflexive saccades (increased number of errors in the AS task and increased number of disinhibitions in the DAS task). Finally, our study did not show significant differences in either response time or error rate between the RAS and the DAS tasks for either control subjects or PD patients. In sum, we report evidence for voluntary saccade execution deficits together with problems inhibiting reflexive saccades in Parkinson's disease patients. These findings were correlated with each other and disease severity, suggesting that eye movement measurement may be a useful tool for studying higher cognitive function. PMID- 16376955 TI - Effects of AP5 infusion into the lateral ventricle on the activities and hippocampal electrical patterns of sleep episodes in rats. AB - Although N-methyl-D-aspartate (NMDA) receptors of the hippocampus are mainly associated with learning and memory that might occur "on-line" during sharp waves (SPWs) and theta-rhythm, the participation of hippocampal NMDA receptors in sleep related processes has not been well studied. In this study, the activity of sleep episodes, hippocampal SPWs and theta-rhythm were recorded in rats received a repeated infusion of NMDA receptor antagonist, D,L-2-amino-5-phosphonopentanoic acid (AP5), into the lateral ventricle in a 5-h daytime sleep. The first trial AP5 infusion (30 mM/2 microl) did not change measures of the activity of slow wave sleep (SWS), paradoxical sleep (PS) and awake episodes, but induced a delay in the latency of the first onset of PS; in the hippocampal EEG, it increased the amplitude of SPWs within SWS and shifted the amplitude/spectral power of theta rhythm from high to low frequency within PS. The repeated AP5 infusion augmented the activity of SWS, and impaired PS and awake episodes; in the EEG-sleep picture, it maintained high scores of SPWs with the complete blockade of theta rhythm generation. When AP5 rat was woken, the theta-rhythm was seen during exploratory behavior. These findings provide evidence that hippocampal NMDA receptors via SPWs or directly associated with the synaptic events of theta rhythm generation are critical for the PS activities. PMID- 16376956 TI - Transgenic barley plants overexpressing a 13-lipoxygenase to modify oxylipin signature. AB - Three chimeric gene constructs were designed comprising the full length cDNA of a lipoxygenase (LOX) from barley (LOX2:Hv:1) including its chloroplast targeting sequence (cTP) under control of either (1) CaMV35S- or (2) polyubiquitin-1 promoter, whereas the third plasmid contains 35S promoter and the cDNA without cTP. Transgenic barley plants overexpressing LOX2:Hv:1 were generated by biolistics of scutella from immature embryos. Transformation frequency for 35S::LOX with or without cTP was in a range known for barley particle bombardment, whereas for Ubi::cTP-LOX no transgenic plants were detected. In general, a high number of green plantlets selected on bialaphos became yellow and finally died either in vitro or after potting. All transgenic plants obtained were phenotypically indistinguishable from wild type plants and all of them set seeds. The corresponding protein (LOX-100) in transgenic T0 and T1 plants accumulated constitutively to similar levels as in the jasmonic acid methyl ester (JAME)-treated wild type plants. Moreover, LOX-100 was clearly detectable immunocytochemically within the chloroplasts of untreated T0 plants containing the LOX-100-cDNA with the chloroplast target sequence. In contrast, an exclusive localization of LOX-100 in the cytoplasm was detectable when the target sequence was removed. In comparison to sorbitol-treated wild type leaves, analysis of oxylipin profiles in T2 progenies showed higher levels of jasmonic acid (JA) for those lines that displayed elevated levels of LOX-100 in the chloroplasts and for those lines that harboured LOX-100 in the cytoplasm, respectively. The studies demonstrate for the first time the constitutive overexpression of a cDNA coding for a 13-LOX in a monocotyledonous species and indicate a link between the occurrence of LOX-100 and senescence. PMID- 16376957 TI - A Kunitz proteinase inhibitor from Archidendron ellipticum seeds: purification, characterization, and kinetic properties. AB - Leguminous plants in the tropical rainforests are a rich source of proteinase inhibitors and this work illustrates isolation of a serine proteinase inhibitor from the seeds of Archidendron ellipticum (AeTI), inhabiting Great Nicobar Island, India. AeTI was purified to homogeneity by acetone and ammonium sulfate fractionation, and ion exchange, size exclusion and reverse phase chromatography (HPLC). SDS-PAGE of AeTI revealed that it is constituted by two polypeptide chains (alpha-chain, M(r) 15,000 and beta-chain, M(r) 5000), the molecular weight being approximately 20 kDa. N-terminal sequence showed high homology with other serine proteinase inhibitors belonging to the Mimosoideae subfamily. Both Native PAGE as well as isoelectric focussing showed four isoinhibitors (pI values of 4.1, 4.55, 5.27 and 5.65). Inhibitory activity of AeTI remained unchanged over a wide range of temperatures (0-60 degrees C) and pH (1-10). The protein inhibited trypsin in the stoichiometric ratio of 1:1, but lacked similar stoichiometry against chymotrypsin. Also, AeTI-trypsin complex was stable to SDS unlike the SDS unstable AeTI-chymotrypsin complex. AeTI, which possessed inhibition constants (K(i)) of 2.46 x 10(-10) and 0.5 x 10(-10)M against trypsin and chymotrypsin activity, respectively, retained over 70% of inhibitory activity after being stored at -20 degrees C for more than a year. Initial studies on the insecticidal properties of AeTI indicate it to be a very potent insect anti-feedant. PMID- 16376958 TI - Oligomeric secoiridoid glucosides from Jasminum abyssinicum. AB - From the root bark of Jasminum abyssinicum (Oleaceae) collected in Congo was isolated tree oligomeric secoiridoid glucosides named craigosides A-C. The three compounds are esters of a cyclopentanoid monoterpene with an iridane skeleton, esterified with three, two and two, respectively, units of oleoside 11-methyl ester. The structures were elucidated by spectroscopic methods and chemical correlations. PMID- 16376959 TI - Sex allocation and dispersal in a heterogeneous two-patch environment. AB - We investigate the evolution of sex allocation and dispersal in a two-habitat environment using a game theoretic analysis. One habitat is of better quality than the other and increased habitat quality influences the competitive ability of offspring in a sex-specific manner. Unlike previous work, we allow incomplete mixing of the population during mating. We discuss three special cases involving the evolution of sex allocation under fixed levels of dispersal between habitats. In these special cases, stable sex-allocation behaviors can be both biased and unbiased. When sex-allocation behavior and dispersal rates co-evolve we identify two basic outcomes. First-when sex-specific differences in the consequences of spatial heterogeneity are large-we predict the evolution of biased sex-allocation behavior in both habitats, with dispersal by males in one direction and dispersal by females in the other direction. Second-when sex-specific differences are small unbiased sex-allocation is predicted with no dispersal between habitats. PMID- 16376960 TI - Proteomic analysis of Myrmecia pilosula (jack jumper) ant venom. AB - Ant sting allergy in Australia is predominantly due to the Myrmecia pilosula species complex. Gel separation of M. pilosula venom is necessary so that the allergenic importance of each component can be defined by western blotting. However, previous PAGE methods produced suboptimal resolution and the components of each band were not precisely defined. Venom was resolved in both non-reduced and reduced form by one-dimensional acid urea PAGE, SDS-PAGE and two-dimensional acid urea-SDS PAGE. Resolved peptides were extracted and analysed by HPLC-MS. Acid urea PAGE and acid urea-SDS PAGE proved more effective than SDS-PAGE for resolution of peptides smaller than 10 kDa. All of the major peptides previously observed in M. pilosula venom were observed in gel resolved venom. Venom was found to primarily consist of peptides with molecular weight <10 kDa, most of which contain disulfide bridges. SDS-PAGE of non-reduced venom clearly defined six higher molecular weight proteins between 26 and 90 kDa. An 8546 Da dimer named pilosulin 5 was observed, but pilosulin 4, a peptide recently proposed to be present in venom was not. A variant of pilosulin 4 here named pilosulin 4.1a, existing as an 8198 Da dimer, was observed and has been characterised. PMID- 16376961 TI - An advanced technique for immuno-labelling of microcystins in cryosectioned cells of Microcystis aeruginosa PCC 7806 (cyanobacteria): implementations of an experiment with varying light scenarios and culture densities. AB - The intracellular localisation of cyanobacterial toxins might well indicate production sites and possible shifts to destination points, thus giving information on possible functions of these toxins within algal cells or at the ecological level beyond. By preparing cells of Microcystis aeruginosa PCC 7806 by cryofixation/cryosectioning and using purified high quality antibodies for immunogold-localisation, excellent ultrastructural integrity and labelling of microcystins was shown. Compared to conventional techniques, including organic solvents, possible dislocation/extraction was significantly minimised, hence, the labelling density was enhanced and the labelling pattern changed. The microcystins were mainly localised within the inner nucleoplasmic area and accumulations of epitopes could be detected around/within intracellular inclusions, such as polyphosphate bodies and carboxysomes. Photosynthetic active radiation (PAR) had a significant effect on microcystin biosynthesis, and the medium light intensity of 25 microE m(-2) s(-1) induced the highest intracellular microcystin contents (up to 160 epitopes per cell and 26 epitopes per microm2). The restriction of the full light spectrum to blue (400-500 nm) or red (>610 nm) wavelengths did not result in any significant effect on microcystin production. However, the subcultures harvested at higher optical densities (>0.5) revealed significantly higher microcystin labelling compared to the less dense cell cultures (OD < 0.5). Altogether, the possibility was discussed whether microcystin might function as an inhibitor of RUBISCO under conditions of C limitations. The effects of light intensity and cell suspension density on intracellular microcystin shown by immuno-detection matched the pattern of microcystin concentrations determined in parallel by HPLC and ELISA. PMID- 16376962 TI - Human papillomaviruses bind a basal extracellular matrix component secreted by keratinocytes which is distinct from a membrane-associated receptor. AB - Human papillomaviruses (HPVs) have previously been shown to adsorb to cultured cells via membrane-associated heparan sulfate (HS) and alpha6 integrin. We demonstrate that cultured keratinocytes uniquely secrete a component into the basal extracellular matrix (ECM) which can function to adsorb HPV particles which can then be internalized by adherent cells. This uncharacterized basal ECM adsorption receptor was secreted by normal human epidermal keratinocytes (NHEK) and by each of the four keratinocyte-derived cell lines we examined, but not by non-keratinocyte cell lines. Multiple HPV types bound preferentially to this keratinocyte-specific receptor over the membrane-associated receptor, and binding to the basal ECM adsorption receptor was refractory to inhibition by heparin. Like the membrane-associated receptor, this basal ECM component was functional as an adsorption receptor in our in vitro infection model using HPV-11. Unlike particle adsorption, however, successful infection with HPV-11 virions remained sensitive to the pretreatment of virions with heparin. The secreted basal ECM receptor did not colocalize with antibodies against HS, perlecan, or alpha6 integrin, but colocalized with antibody against laminin-5, a marker of keratinocyte ECM and an abundant component of the basement membrane in mucosa and skin. These findings suggest a model for natural infections in which HPV virions, nonspecifically adsorbed to HS on suprabasal keratinocytes throughout an epithelial wound, might be transferred to mitotically active migrating keratinocytes via an intermediate association with the ECM secreted by these cells as they reestablish the basement membrane. PMID- 16376963 TI - A common mechanism for perceptual filling-in and motion-induced blindness. AB - Perceptual-filling-in (PFI) and motion-induced-blindness (MIB) are two phenomena of temporary blindness in which, after prolonged viewing, perceptually salient targets repeatedly disappear and reappear, amidst a field of distracters (i.e., non-targets). Past studies have shown that boundary adaptation is important in PFI, and that depth ordering between target and distracter pattern is important in MIB. Here we show that the reverse is also true; that boundary adaptation is important in MIB, and that depth ordering is important in PFI. Results corroborate our earlier conjecture that PFI and MIB are highly related phenomena that share a common underlying mechanism. We argue that this mechanism involves boundary adaptation, but also that the depth effect shows that boundary adaptation can be no more than a sufficient cause of PFI and MIB, and not a necessary one. PMID- 16376964 TI - Absorbability of estrone and 17beta-estradiol in water onto activated carbon. AB - With a view to reducing pollution in the aquatic environment, estrone (E1) and 17beta-estradiol (E2) were tested for their absorbability in water onto activated carbons (ACs) with various pore-size distributions. In batch-type adsorption measurements, all adsorption isotherms obtained were found to fit a Freundlich equation. In case of eight different kinds of commercial AC in pure water, the amount adsorbed at equilibrium concentration of 1 microg/L was in the range of 25.6-73.5mg/g for E1 and 21.3-67.6 mg/g for E2. In case of pre-used ACs in water sand-filtered for use as drinking water, the amount adsorbed at 1 microg/L was in the range of 3.5-8.2mg/g for E2. In the case of two commercial ACs in river water and in effluent from secondary clarifier at municipal sewage treatment plant (MSTP), both originally containing E2, was in the range of 0.1-0.2 and 0.3-1 microg/g, respectively, at 1 ng/L. The difference of amount adsorbed onto AC was discussed in view of hydrophobicity of target chemicals, specific surface area and mean pore diameter of AC, and the difference in the absorbability and the ratios of co-present substances for the target compound. PMID- 16376965 TI - Oxidation of resin and fatty acids by ozone: kinetics and toxicity study. AB - The ozonation of resin and fatty acids (RFAs) found in pulp mill effluents was investigated using rapid-scan stopped-flow spectrophotometry. RFAs oxidation (i.e., degradation) efficiency increased with increasing the amount of used ozone and temperature. The degradation process with respect to the acid was found to follow first-order kinetics. The ozonation of RFAs was modeled as an overall second-order reaction for both reactants. The apparent overall second-order rate constants were calculated based on the pseudo first-order rate constants obtained from the kinetic data fitting for the acid degradation. The apparent overall second-order rate constant was affected by pH and temperature. At 20 degrees C and when pH increased from 8 to 11, the apparent overall second-order rate constant increased almost by a factor of 5 (from 3.9 x10(3) to 1.8 x10(4)M(-1)s( 1)) for 9 mgL(-1) resin acid and a factor of 4 (from 9.6 x10(3) to 3.9 x10(4)M( 1)s(-1)) for 8 mgL(-1) fatty acid. At pH 8 and as temperature increased from 10 to 20 degrees C, the apparent overall second-order rate constant increased almost by a factor of 5 (from 8.2 x10(2) to 3.9 x10(3)M(-1)s(-1)) for 9 mgL(-1) resin acid and a factor of 3 (from 3.5 x10(3) to 9.6 x10(3)M(-1)s(-1)) for 8 mgL(-1) fatty acid. Microtox bioassay tests were completed to evaluate the toxicity of RFAs samples before and after ozonation. For the resin acid, there was an increase in toxicity as a result of ozonation. Meanwhile, toxicity of fatty acid samples decreased as a result of ozonation. PMID- 16376966 TI - Biological activation of carbon filters. AB - To prepare biological activated carbon (BAC), raw surface water was circulated through granular activated carbon (GAC) beds. Biological activity of carbon filters was initiated after about 6 months of filter operation and was confirmed by two methods: measurement of the amount of biomass attached to the carbon and by the fluorescein diacetate (FDA) test. The effect of carbon pre-washing on WG 12 carbon properties was also studied. For this purpose, the nitrogen adsorption isotherms at 77K and Fourier transform-infrared (FT-IR) spectra analyses were performed. Moreover, iodine number, decolorizing power and adsorption properties of carbon in relation to phenol were studied. Analysis of the results revealed that after WG-12 carbon pre-washing its BET surface increased a little, the pH value of the carbon water extract decreased from 11.0 to 9.4, decolorizing power remained at the same level, and the iodine number and phenol adsorption rate increased. In preliminary studies of the ozonation-biofiltration process, a model phenol solution with concentration of approximately 10mg/l was applied. During the ozonation process a dose of 1.64 mg O(3)/mg TOC (total organic carbon) was employed and the contact time was 5 min. Four empty bed contact times (EBCTs) in the range of 2.4-24.0 min were used in the biofiltration experiment. The effectiveness of purification was measured by the following parameters: chemical oxygen demand (COD(Mn)), TOC, phenol concentration and UV(254)-absorbance. The parameters were found to decrease with EBCT. PMID- 16376967 TI - Study of catalyzed ozonation for advanced treatment of pulp and paper mill effluents. AB - Ozonation and catalytic ozonation (TOCCATA process) were used as tertiary treatments of wastewaters from three different pulp and paper mills. Laboratory batch experiments were conducted to assess the efficiency of each oxidation system for removal of organic matter. The investigations measured ozone consumption rate, variations in chemical oxygen demand (COD), total organic carbon (TOC), suspended solids (SS) and molecular weight distribution with contact time. For conventional ozonation, ozone consumption rate was dependent on the nature of the effluent. Organic matter elimination occurred both by oxidation and precipitation. Precipitation played a major role on TOC removal varying with the effluent, and was responsible for production of high final SS concentrations. However, the effluent type did not affect the ozone consumption rate for TOCCATA catalyzed reactions. Using TOCCATA, it was shown that organic matter was removed through steady conversion of organic carbon to carbon dioxide. Finally the two oxidation systems were compared with respect to their impact on molecular weight distribution. A total removal of the two initial fractions of compounds (high and low molecular weights) was observed with two effluents. With the third effluent, only the initial fraction of low molecular weight compounds was removed by the two oxidizing systems. The results showed that ozonation and TOCCATA-catalyzed ozonation could achieve removals of COD of 36-76%. Depending on the effluent type, the amount of ozone consumed per gram of COD removed was lower for conventional or for catalytic ozonation. PMID- 16376968 TI - Distribution and sources of hydrocarbons in surface sediments of Gemlik Bay (Marmara Sea, Turkey). AB - Seabottom sediments from Gemlik Bay, one of the most polluted spots in SW Marmara Sea, were analyzed for parent polycyclic aromatic hydrocarbons (PAHs) using gas chromatography-mass spectrometry. The concentration of 14 PAH compounds in sediment samples collected from 61 locations are distributed in a broad spectrum from low to very high concentration levels (50.8-13482 ng g-1). No significant correlation was found between summation operatorPAHs and organic carbon content while summation operatorPAHs increase slightly with silt/clay ratio. Therefore the distribution and concentrations of PAHs would be determined more by direct input, rather than by the type of sediment found locally. The most polluted areas are distributed nearshore eastern (Gemlik) and southern (Kursunlu, Mudanya and Trilye) coasts which are mainly influenced by rapid ecotourism development, direct discharges from rivers, surface run-off and drainage from port areas, domestic and industrial effluent discharges through outfalls and various contaminants from ships. Special PAH compound ratios, such as Phe/Anth, Flu/Py, B[a]A/Chry; LMWPAH/HMWPAH; Per/; Per/summation operatorPAH; Per/summation operator(penta-aromatics) and Flu/(Py+Flu), were calculated to evaluate different hydrocarbon origins and their relative importance. Pyrolytic activity is dominant along the highly-populated eastern and southern coasts. Meanwhile, petrogenic activity mixed with pyrolytic activity is a matter of fact in front of the main industrial-tourism ports and anchoring areas as well. Higher concentration of perylene are distributed along the mostly polluted eastern and southern coastal areas, however, the concentrations of perylene relative to the penta-aromatic isomers are dominant especially in the northern and deepest sectors of the bay, indicating diagenetic origin for the presence of perylene. PMID- 16376969 TI - Estimation of 1999-2004 dietary daily intake of PCDDs, PCDFs and dioxin-like PCBs by a total diet study in metropolitan Tokyo, Japan. AB - A study of the dietary intake of dioxins, consisting of polychlorinated dibenzo-p dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dioxin-like PCBs) through foods retailed in the metropolitan Tokyo area from 1999 to 2004 was carried out by the total diet market basket method on the basis of food classification (14 groups) and the data on food consumption in the Tokyo region obtained from the Japan Nutrition Survey. The daily intake of dioxins per kg of body weight for a 50 kg average adult body was 2.18 pg TEQ/kg/day in 1999, 1.87 pg TEQ/kg/day in 2000, 1.25 pg TEQ/kg/day in 2001, 1.60 pg TEQ/kg/day in 2002 and 2003 and 1.55 pg TEQ/kg/day in 2004, respectively. These amounts were less than the tolerable daily intake (TDI) of 4 pg TEQ/kg/day for dioxins established in Japan. The dioxins taken daily through fish and shellfish (group 10) accounted for more than 50% of sum WHO-TEQs. In addition, more than 90% of the daily intake of dioxins was taken through fish and shellfish (group 10), meat and eggs (group 11), milk and dairy products (group 12). Also, this study clearly showed that the ratio of dioxin-like PCBs in the daily intake of dioxins was increasing yearly because the reduction rate of dioxin-like PCBs was lower than that of PCDDs and PCDFs in foods. PMID- 16376970 TI - Mercury in sediments and Nassarius reticulatus (Gastropoda Prosobranchia) in the southern Venice Lagoon. AB - The southern basin of the Venice Lagoon has been the focus of fewer studies concerning contamination from heavy metals than the northern and central basins. A recent increase in urban waste waters from Chioggia town, as well as dockyards, shipping and fishing activities, affect this part of the lagoon. The aim of this study was to investigate the total mercury (THg) incidence in sediments and Nassarius reticulatus gastropods in order to assess its distribution and evaluate the level of contamination. THg concentration measured in bottom sediments ranged between 0.1 and 3.4 mg/kg d. wt. The enrichment factor (EF) showed high values (avg. 30, max 49) near the dockyards of Chioggia; the lowest (avg. 9, max 17) were found in the coastal marine sediments near the port entrance of the southern basin. THg in marine scavenger gastropods accumulated in N. reticulatus with concentrations falling within the range of 0.3-1.3 mg/kg d. wt. A positive correlation was found between THg concentration in sediments and in N. reticulatus in all sites, excluding the dockyards. A first local cause for mercury pollution might be attributed to the antifouling paints used in great quantity in the recent past near the town of Chioggia. Moreover, fine suspended sediments associated with tidal flushing are suggested as possibly being the vehicle for pollutant dispersal from the Marghera industrial area to the whole of Venice's lagoon. PMID- 16376971 TI - Quantification and speciation of mercury in soils from the Tripui Ecological Station, Minas Gerais, Brazil. AB - Contents of total mercury, organic carbon, total sulfur, iron, aluminum and grain size and clay mineralogy were used along with Pearson's correlation and Hg thermal desorption technique to investigate the presence, distribution and binding behavior of Hg in soils from three depths from the Tripui Ecological Station, located near Ouro Preto, Minas Gerais State, Brazil. The soils studied had predominantly medium and fine sand texture (0.59-0.062 mm), acid character and Hg contents ranging from 0.09 to 1.23 microg/g. The granulometric distribution revealed that Hg is associated with coarse sand (2-0.59 mm) and silt and clay (<0.062 mm) and presents similar Hg concentrations in both fractions. Mercury distribution in soil profiles showed that Hg was homogeneously distributed throughout the depths at most sites. Hg thermal desorption curves show that mercury occurs not only as Hg2+ predominantly bound to organic components in most of the samples, but also in the form of cinnabar in some. Pearson's correlation confirmed that mercury is associated with organic matter and sulfur and possibly with sulfur-bearing organic matter in most samples. PMID- 16376972 TI - Occurrence and hazard screening of alkyl sulfates and alkyl ethoxysulfates in river sediments. AB - Alkyl sulfates (AS) and alkyl ethoxysulfates (AES) are High Production Volume (HPV) 'down-the-drain' chemicals widely used globally in detergent and personal care products, resulting in low levels (ng to microg L(-1) range) ultimately released to the environment via wastewater. These surfactants have a strong affinity for sorption to sediments. However, data regarding the fate and effects following release into the environment has not been reported. Sediment samples from both normal exposed and presumably low exposed locations (background) were analyzed to determine the levels of AS/AES. The method used in this study shows broad applicability across various sediment types and the most common congeners of AS/AES. The combined levels of AS/AES detected in the two presumed lower exposed sites ranged from 0.025 and 0.034 microg g(-1) on a dry weight (dw) basis while the presumed higher exposed site had combined levels of AS/AES of 0.117 microg g(-1) (dw) based on triplicate analyses. Results indicate that detectable levels of AS/AES can be found in sediments in the environment at these three sites that are below the concentrations expected to produce significant adverse ecological effects for individual homologues and the whole mixture, the hazard screening for these three sites had PEC(porewater)/PNEC(total mixture) ratios of 0.007-0.024. However, further investigation of potential effects and risk assessment is warranted. PMID- 16376975 TI - Mast cell density, angiogenesis, and their significance in tumor development. PMID- 16376976 TI - Primary lymphoma of the uterine cervix: An approach to management. PMID- 16376977 TI - A randomized intervention study of sun protection promotion in well-child care. AB - BACKGROUND: This study evaluated the behavioral impact of a skin cancer prevention program in which health care providers delivered advice and materials to parents of infants over a 3-year period from 1998 to 2001. METHODS: Fourteen offices of a large managed care organization in Colorado were randomly assigned to the intervention or control groups. 728 infants and their parents were recruited within 6 months of birth. At intervention offices, health care providers attended orientation sessions, prompts for delivering sun protection advice were placed in medical records, and parents received sun protection packets at each well-child visit between 2 and 36 months of age. RESULTS: Based on provider self-report and exit interviews of parents, providers in the intervention group delivered approximately twice as much sun protection advice as providers in the control group. Annual telephone interviews of parents indicated small but statistically significant differences in parent sun protection practices favoring the intervention. Skin exams revealed no significant differences in tanning, freckling, or number of nevi. Behavioral differences between groups appeared to grow over the 3 years of follow-up. CONCLUSIONS: This intervention strategy was successful in increasing the delivery of sun protection advice by health care providers and resulted in changes in parents' behaviors. While the behavioral effect was probably not strong enough to reduce risk for skin cancer, the effect may increase as children age and have more opportunities for overexposure to the sun. PMID- 16376978 TI - Metabolic syndrome in Japanese men and women with special reference to the anthropometric criteria for the assessment of obesity: Proposal to use the waist to-height ratio. AB - BACKGROUND: Obesity is one of the criteria for defining metabolic syndrome. However, overt obesity in Asians is relatively low despite high prevalence of metabolic risks. METHODS: We investigated the effectiveness of various anthropometric indices {body mass index (BMI), waist circumference, waist-to height ratio (W/Ht)} for the evaluation of coronary risk factors (hypertension, hyperglycemia, hypertriglyceridemia, and low HDL cholesterol) and as one of the criteria for metabolic syndrome (clustering of three or more from one obesity and four coronary risk factors) in Japanese on 6141 men and 2137 women. RESULTS: (1) The areas under the receiver operating characteristic curves for the different anthropometric indices to identify any one and two or more coronary risk factors were highest for W/Ht. (2) The sensitivities for the identification of any one and two or more coronary risk factors were greater for W/Ht > or =0.5 than BMI > or =25, > or =23, and indices of waist circumference (Adult Treatment Panel III and Japan Society for the Study of Obesity). (3) The prevalence of metabolic syndrome varied greatly by different anthropometric indices, and the percentages of obesity risk factors in metabolic syndrome were highest for W/Ht > or =0.5 in both genders (approximate 95%). CONCLUSIONS: W/Ht >/=0.5 may be the most effective anthropometric index for screening Japanese people for metabolic syndrome. PMID- 16376979 TI - Effect of 4 weeks of Pilates on the body composition of young girls. AB - BACKGROUND: There is a need to find ways to increase the physical activity levels and improve the body composition and blood pressure of girls. METHODS: Thirty 11 year-old girls were recruited from two after school programs in Houston Texas in Spring 2005. Participants from one program (16) were randomly assigned to intervention, the other (14) served as controls. BMI, BMI percentile, waist circumference and blood pressure were assessed before and after the intervention. Pilates classes were provided free of charge for an hour per day at the intervention site, 5 days a week, for 4 weeks. Four participants wore heart rate monitors during every session and completed enjoyment and perceived exertion questionnaires. Repeated measures analysis of variance with time (within) and group (between) as factors was performed. RESULTS: Mean attendance was 75%, mean heart rate 104 bpm, mean perceived exertion 5.9 (1-10 scale) and enjoyment 4.4 (1 5 scale). There was a significant (P = 0.039) time by group interaction for BMI percentile. Graphs indicated that this difference was influenced by large reductions in the BMI percentile of healthy girls. CONCLUSIONS: Girls enjoyed Pilates, and participation for 4 weeks lowered BMI percentile. Pilates holds promise as a means of reducing obesity. PMID- 16376980 TI - Birth of live Spanish ibex (Capra pyrenaica hispanica) derived from artificial insemination with epididymal spermatozoa retrieved after death. AB - As a consequence of increasing limitations to maintaining genetic variability in endangered wildlife species, methods of assisted reproduction widely used in domestic animals are being applied to nondomestic species. However, practical efforts have met limited success to date. The Spanish ibex (Capra pyrenaica hispanica) is a wild caprine originating exclusively in the mountains of Spain. This study was designed to evaluate the fertilizing capability of cryopreserved Spanish ibex epididymal spermatozoa recovered postmortem. For this purpose, we have previously evaluated the effect of time elapsed between death and sperm recovery on spermatic parameters, and the fertilization ability of frozen-thawed spermatozoa using heterologous in vivo fertilization by intrauterine insemination in domestic goat (Capra hircus). The time of death significantly affected most sperm quality parameters (motility, viability and intact acrosomes). The fertility obtained by heterologous artificial insemination was 18.7%, and only goats inseminated with spermatozoa recovered within 8h after death became pregnant. Our findings showed that heterologous in vivo fertilization is a useful method to evaluate the fertilizing capacity of sperm samples in rare or wild species. Sperm samples, with verified fertilization ability in the previous trial, were used to inseminate a total of six ibex females. Inseminations resulted in one pregnancy. The study demonstrated for the first time the feasibility of applying artificial insemination in Spanish ibex. PMID- 16376981 TI - Microtensile bond strength of different components of core veneered all-ceramic restorations. Part II: Zirconia veneering ceramics. AB - OBJECTIVES: The purpose of this research was to investigate microtensile bond strength between one type of zirconia substrate and various commercial veneer ceramics. The effect of an optional liner material between the core and veneer was also evaluated where applicable. METHODS: Bilayered zirconia veneer discs were fabricated from five layering and two pressable veneer ceramics. Additionally, discs from each veneer ceramic were prepared. The discs were cut into microbars of 6 mm in length and 1mm in cross-section. The microtensile bond strength was tested in a universal testing machine. The fracture surfaces of the microbars were examined with scanning electron microscopy (SEM) and EDAX. RESULTS: The microtensile strength of Rondo Dentine and Lava Dentine veneer ceramics were significantly higher than the other tested veneer ceramics. Furthermore, the layered systems Rondo Dentine and Ceram Express were significantly stronger than the other tested core-veneer ceramics. The application of liner material dramatically affected the bond strength and failure mode, which was also material dependent. SEM analysis showed that two pressable veneers and one type of layering veneer ceramic failed entirely cohesively in veneer side while the remaining test groups had higher percentage of interfacial failure. SIGNIFICANCE: Selection of stronger veneer ceramics which have good bond strength with zirconia can reduce the chances of chipping and delamination under function. The liner material should only be used with some layering veneers but not in combination with pressable veneers as it will result in weakening of the microtensile bond strength. PMID- 16376982 TI - Involvement of oxidative stress in mutagenicity and apoptosis caused by dental resin monomers in cell cultures. AB - OBJECTIVE: This investigation studied the possibility that apoptosis as well as mutagenicity induced by resin monomers are mediated by oxidative stress. METHODS: A range of dilutions of three resin monomers (GMA, TEGDMA, and HEMA) was added to culture medium (DMEM/10% FBS), of V79-4 fibroblasts and RPC-C2A pulp cells for 24 h. Their cytotoxic effects were measured by a colorimetric functional assay (MTT). Chromosomal aberration induced by the resin monomers was investigated by counting micronuclei in V79-4 cells. The effects of the resin monomers on DNA fragmentation were viewed by agarose gel electrophoresis of DNA, isolated from RPC-C2A pulp cells that were treated by resin compounds. Resin monomer-induced apoptosis was further confirmed by flow cytometry (staining with both annexin V FITC and PI). RESULTS: All monomers exhibited a dose-dependent cytotoxic effect, and the ranking of the cytotoxicity based on TC50 was GMA > TEGDMA > HEMA. The resin monomer-induced cytotoxicity was significantly decreased by co-treatment with N-acetylcystein (NAC), an antioxidant. The authors also confirmed a dose dependent genotoxicity of the resin monomers that had induced micronucleated cells in V79-4 fibroblasts. Similar to the effects on cytotoxicity, NAC reduced the numbers of micronuclei in comparison with those generated by the resin monomers. The preventive effects of NAC were also observed in monomer-induced apoptosis in RPC-C2A cells. A DNA ladder pattern, characteristic of apoptosis, was shown at cytotoxic concentrations, but NAC blocked the resin monomer-mediated DNA fragmentation. The preventive effects of NAC on apoptosis were confirmed by Annexin V staining. Cells exposed to 300 microM GMA, 7 mM TEGDMA, or 14 mM HEMA for 24 h showed a significant increase in apoptotic cells, while NAC co-treatment caused a reduction in apoptotic cells compared to controls. SIGNIFICANCE: These findings suggest that glutathione depletion and oxidative stress are responsible for GMA, TEGDMA, and HEMA-induced mutagenicity and apoptosis. PMID- 16376983 TI - Effect of pontic height on the fracture strength of reinforced interim fixed partial dentures. AB - OBJECTIVES: This in vitro study evaluated the fracture load of interim FPDs made with various materials and pontic heights. The hypothesis was that different materials and pontic heights result in different fracture resistance. METHODS: Groups of interim FPDs were fabricated with prosthodontic resin materials on two abutments with two different pontic heights (4.3 and 5.8 mm) and a pontic width of 4 units (19 mm) (n = 3). The following materials were tested: (1) a thermoplastic polymer (Promysan Star), (2) Promysan Star with a veneering composite (Vita Zeta), (3) a non-impregnated polyethylene fiber reinforced resin (Ribbond) with a veneering composite (Sinfony), (4) an impregnated fiber reinforced composite system (Targis/Vectris), and (5) a conventional poly methyl methacrylate (PMMA) (Biodent K+B, control group). After 5000 thermocycles, the FPDs were temporarily fixed with a provisional cement on the corresponding abutments and tested for fracture strength. One-way and two-way ANOVA and Bonferroni-Dunn's multiple comparison tests were performed for the statistical analysis (alpha = 0.05). RESULTS: The mean fracture strength ranged from 83.0 to 625.9 N for a pontic height of 4.3 mm and from 97.2 to 893.7 N for a pontic height of 5.8 mm. Vectris/Targis FPDs of both pontic heights exhibited significantly superior fracture resistance compared to the corresponding Promysan, Promysan/Vita Zeta, Ribbond/Sinfony and Biodent groups. Except Biodent FPDs, fracture resistance of FPDs with a pontic height of 4.3 mm showed no significant differences compared to a pontic height of 5.8 mm for each material. SIGNIFICANCE: Material type of the FPDs has a significant influence on the fracture strength, whereas pontic height has no significant effect (except control group). PMID- 16376985 TI - Enhanced basal apoptosis in cultured term human cytotrophoblasts is associated with a higher expression and physical interaction of p53 and Bak. AB - We tested the hypothesis that the expression levels of p53 and the pro-apoptotic mediators from the Bcl-2 family are higher in cytotrophoblasts, when compared to cultures with abundant syncytiotrophoblasts. Cytotrophoblasts isolated from normal term human placentas were cultured in Dulbecco's Modified Eagle medium (DMEM) for 24 h, when the cytotrophoblast phenotype predominates, in DMEM for 72 h, when the syncytiotrophoblast phenotype predominates, or in Ham's-Waymouth medium or DMEM with 1.5% dimethylsulfoxide, each of which maintains the cytotrophoblast phenotype through 72 h of culture. Apoptosis was assessed by detection of cleavage products of poly-ADP-ribose polymerase, by expression of cleaved cytokeratin 18 intermediate filaments, and by assessment of caspase-3 activity. Independent of time in culture, cytotrophoblasts showed higher levels of apoptosis compared to syncytiotrophoblasts. Cytotrophoblasts also expressed a 2-fold higher level of p53, a 2-fold lower level of 60 kDa Mdm-2 protein, a 2 fold higher level of Bak, but no differences in the expression of 90 kDa Mdm-2, Bcl-2, Bcl-X(L), Mcl-1, Bax, Bad, and Bad phosphorylated at the serine(112), serine(136), or serine(155) sites, compared to the syncytiotrophoblasts. Using co immunoprecipitation, we demonstrated a greater degree of Bak-p53 interaction in cytotrophoblasts than in syncytiotrophoblasts. We also detected Bak-Mcl-1 interaction that was no different between the two phenotypes. Among the proteins studied, enhanced p53 activity, differential Bak expression, and Bak-p53 interactions may contribute to the higher level of constitutive apoptosis in cultures of cytotrophoblasts compared to syncytiotrophoblasts. PMID- 16376986 TI - Tumour necrosis factor-alpha converting enzyme in human gestational tissues from pregnancies complicated by chorioamnionitis. AB - Chorioamnionitis increases the risk of preterm labour and is associated with adverse neonatal outcomes including cerebral palsy. Tumour necrosis factor-alpha (TNF-alpha) derived from the gestational tissues (placenta, fetal membranes and maternal decidua) is thought to play a pivotal role in the induction of cytokine response in chorioamnionitis. Tumour necrosis factor-alpha converting enzyme (TACE) is essential for the release of TNF-alpha. Our aim was to determine whether the expression of TACE is increased in human gestational tissues from pregnancies complicated by chorioamnionitis, and whether lipopolysaccharide (LPS) causes increased expression of TACE in the human gestational tissues in vitro. The immunostaining of TACE was generally more intense, in particular in the syncytiotrophoblast and stromal cells, in villous samples from pregnancies complicated by chorioamnionitis than those from normal pregnancies. Increased immunoreactivity of TACE was also noted in the amnion and choriodecidua. In parallel, there was an increased infiltration of monocytes/macrophages within the villous stroma and choriodecidua. As a complement to our in vivo findings, LPS significantly increased the levels of mRNA and protein of TACE in a dose dependent response in villous and fetal membrane explant cultures. Together, our results imply a potential role of TACE in the pathogenesis of chorioamnionitis. PMID- 16376984 TI - Surface-immobilization of adhesion peptides on substrate for ex vivo expansion of cryopreserved umbilical cord blood CD34+ cells. AB - The interaction between integrins and extracellular matrix proteins play an important role in the regulation of hematopoiesis. Human hematopoietic progenitor cells express very late antigen-4 (VLA-4) and VLA-5, which mediate their interaction with fibronectin by recognizing the connecting segment-1 (CS-1 and RGD motifs, respectively. In this study, we investigated the ex vivo expansion of human umbilical cord blood (UCB) CD34+ cells on synthetic substrates surface immobilized with peptides containing the CS-1 binding motif (EILDVPST) and the RGD motif (GRGDSPC). These peptides were covalently conjugated to poly(ethylene terephthalate) (PET) film at a surface density of 2.0-2.3 nmol/cm2. UCB CD34+ cells were cultured for 10 days in serum-free medium supplemented with recombinant human thrombopoietin, stem cell factor, flt3-ligand and interleukin 3. The highest cell expansion fold was observed on the CS-1 peptide-modified surface, where total nucleated cells, total colony forming unit, and long-term culture initiating cells were expanded by 589.6+/-58.6 (mean+/-s.d.), 76.5+/-8.8, and 3.2+/-0.9-fold, respectively, compared to unexpanded cells. All substrates surface-immobilized with peptides, including the control peptides, were more efficient in supporting the expansion of CD34+, CFU-GEMM and LTC-ICs than tissue culture polystyrene surface. Nevertheless, after 10-days of ex vivo expansion from 600 CD34+ cells, only cells cultured on CS-1-immobilized surface yielded positive engraftment, even though the frequency was low. PET surface immobilized with RGD peptide was less efficient than that with CS-1 peptide. Our results suggest that covalently immobilized adhesion peptides can significantly influence the proliferation characteristics of cultured UCB CD34+ cells. PMID- 16376987 TI - Defining the role of gene regulation in resistance to HDAC inhibitors--mechanisms beyond P-glycoprotein. AB - The manuscript by Yamada et al. "Depsipeptide-resistant KU812 cells show reversible P-glycoprotein expression, hyperacetylated histones, and modulated gene expression profile," describes not only development of the major resistance mechanism, P-glycoprotein expression, but how histone deacetylase inhibitors can modulate the gene expression profile to promote survival and resistance to depsipeptide. PMID- 16376988 TI - Functional analysis of two putative chromosomal replication origins from Pseudomonas aeruginosa. AB - Two autonomously replicating elements previously isolated from Pseudomonas aeruginosa were characterized in vitro for pre-priming complex formation using combinations of replication proteins from P. aeruginosa and Escherichia coli. The results of these studies showed that the P. aeruginosa DnaA and DnaB proteins could form a pre-priming complex on plasmid templates containing either of the two autonomously replicating elements of P. aeruginosa, pYJ50 (containing oriCI), and pYJ52 (containing oriCII), or the E. coli chromosomal origin (plasmid pYJ2). The E. coli DnaA, DnaB, and DnaC proteins were also able to form a pre-priming complex on pYJ2, pYJ50, and pYJ52. Neither pYJ50 nor pYJ52 could be established in E. coli, suggesting a block in steps subsequent to the formation of the pre priming complex. Similarly, pYJ2 could not be established in P. aeruginosa. Since pYJ50 and pYJ52 could be established in P. aeruginosa and both putative origins form a pre-priming complex in vitro, attempts were made to delete each of these two putative origins. The results indicate that the oriCI sequence is essential for cell viability under typical laboratory growth conditions but that oriCII is not. PMID- 16376989 TI - Use of plant genotoxicity bioassay for the evaluation of efficiency of algal biofilters in bioremediation of toxic industrial effluent. AB - The toxicity and efficacy of an algal-based bioremediation technology were assessed through bioassays for ecological risk of contaminated industrial effluents. The algal bioremoval of heavy metals was evaluated using an in vitro approach. Phytogenotoxicity tests were conducted with Allium cepa and Vicia faba plants to evaluate the genotoxicity of the industrial effluents before and after treatment with different kinds of algal biofilters (BF). Root cells were exposed for 24 h to different dilutions of both raw and treated effluent of a chemical fertilizer factory. Three cytogenetic endpoints were used to assess the mutagenic potencies of the industrial effluent: mitotic inhibition, mitotic chromosome aberrations, and nuclear irregularities in interphase cells. Before algal treatment, the industrial effluent caused strong genotoxic effects represented by severe inhibition in mitotic activity of meristematic cells and high frequency of both chromosome and nucleus abnormalities. After algal treatment, the cytotoxic effects of 30% and 60% concentrations of the treated effluent were comparable to those of 5% and 10% concentrations before treatment, respectively, and the frequency of both chromosome and nuclear abnormalities declined by approximately 50%. Statistical analysis of the data indicates a significant reduction in genotoxicity associated with a remarkable reduction in heavy metal concentrations after bioremediation by algal BF. The Allium and Vicia genotoxicity approach was effective in monitoring bioremediated effluent for toxicity. PMID- 16376990 TI - Chlamydia trachomatis occurrence and its impact on sperm quality in chronic prostatitis patients. AB - OBJECTIVES: The role of Chlamydia trachomatis (CT) in the pathogenesis of chronic prostatitis and its impact on male fertility remain controversial. The aim of this study was to assess the occurrence of chlamydial infection in chronic prostatitis patients and its impact on semen quality. METHODS: Urine and semen samples were assayed for the presence of microbial infection. CT-specific IgG and IgA antibodies were measured in serum and seminal plasma. Semen parameter analysis, anti-sperm antibody determinations and inflammatory cytokines measurements were performed. RESULTS: CT was detected in 10% of semen from chronic prostatitis patients. CT-specific IgG and IgA were found in 7.5% and 32.5% of the seminal plasma and in 15.0% and 2.5% of the serum samples from patients. Most of the patients that evidenced CT infection also evidenced CT specific antibodies either in semen or in serum. We found that chlamydial infection has no detrimental effects on sperm quality. We neither found abnormal levels of serum PSA nor of seminal inflammatory cytokines in CT-infected patients. CONCLUSIONS: Our results support the potential role of CT in chronic prostatitis, its importance in diagnosis and that this infection does not seriously compromise sperm quality. PMID- 16376991 TI - Online measurement of motivational processes: introducing the Continuous Delay Aversion Test (ConDAT). AB - The Continuous Delay Aversion Test (ConDAT), a new computer task for online monitoring and continuously measuring delay aversion (DA), is introduced. DA is a motivational style related to a shortened delay gradient which is proposed as a major endophenotype of attention deficit hyperactivity disorder (ADHD). It is characterised by avoiding or escaping from delay-rich situations despite the prospects of a reward. In each ConDAT trial the rapidly diminishing reward/delay ratio, which tends asymptotically towards zero, is visually presented on the computer screen. The test subject is permanently confronted with the question whether to quit or to continue the trial in the face of the deteriorating reward/time ratio. An elaborated control of stimuli and responses, including the sending of trigger codes to external recording devices, makes the task useful for neurophysiological or brain imaging experiments. Compared to existing tasks, the ConDAT is more flexible and sensitive due to its asymptotic open-ended trials and the interval-scaled output measure. Pilot data give evidence for satisfactory reliability and external validity of the task. PMID- 16376992 TI - Errors in the estimation of the variance: implications for multiple-probability fluctuation analysis. AB - Synapses play a crucial role in information processing in the brain. Amplitude fluctuations of synaptic responses can be used to extract information about the mechanisms underlying synaptic transmission and its modulation. In particular, multiple-probability fluctuation analysis can be used to estimate the number of functional release sites, the mean probability of release and the amplitude of the mean quantal response from fits of the relationship between the variance and mean amplitude of postsynaptic responses, recorded at different probabilities. To determine these quantal parameters, calculate their uncertainties and the goodness-of-fit of the model, it is important to weight the contribution of each data point in the fitting procedure. We therefore investigated the errors associated with measuring the variance by determining the best estimators of the variance of the variance and have used simulations of synaptic transmission to test their accuracy and reliability under different experimental conditions. For central synapses, which generally have a low number of release sites, the amplitude distribution of synaptic responses is not normal, thus the use of a theoretical variance of the variance based on the normal assumption is not a good approximation. However, appropriate estimators can be derived for the population and for limited sample sizes using a more general expression that involves higher moments and introducing unbiased estimators based on the h-statistics. Our results are likely to be relevant for various applications of fluctuation analysis when few channels or release sites are present. PMID- 16376993 TI - Delayed-onset PTSD: a prospective study of injury survivors. AB - BACKGROUND: Recent studies have indicated that delayed-onset posttraumatic stress disorder (PTSD) (i.e., the development of PTSD more than 6 months posttrauma) is generally characterised by subsyndromal diagnoses within the first 6 months. This study sought to examine the relationship between sub-clinical levels of PTSD symptoms at 3 months posttrauma and delayed onset PTSD at 12 months in a large sample of traumatic injury survivors. METHODS: Three hundred and one consecutively admitted injury survivors were assessed at 3 and 12 months posttrauma. PTSD was diagnosed according to DSM-IV criteria, while partial and subsyndromal diagnoses were based on recent definitions developed by Mylle and Maes [Mylle, J., Maes, M., 2004. Partial posttraumatic stress disorder revisited. J. Affect. Disord. 78, 37-48]. RESULTS: Eight percent of participants was diagnosed with 3-month PTSD while 10% was diagnosed with 12-month PTSD. Nearly half (47%) of 12-month PTSD cases were of delayed onset. The majority of those with delayed-onset were diagnosed with partial or subsyndromal PTSD at 3 months. Ten percent of delayed onset cases did not meet partial or subsyndromal criteria. LIMITATIONS: As symptoms were not assessed at 6 months (the DSM cut-off for delayed PTSD), it could not be conclusively determined that delayed-onset cases had not developed PTSD between 3 and 6 months posttrauma. CONCLUSION: A considerable proportion of 12-month PTSD diagnoses was delayed in onset. While most demonstrated 3-month morbidity in the form of partial and subsyndromal diagnoses, a minority did not. Thus, clinicians should consider subthreshold diagnoses as potential risk factors for delayed-onset PTSD. Future research is required to identify factors that may predict delayed-onset PTSD in trauma survivors without evidence of prior PTSD pathology. PMID- 16376994 TI - Odor perception in patients with mood disorders. AB - BACKGROUND: Olfaction has obvious correlates with emotional processes but little is known about the several aspects of olfaction in psychiatric disorders characterized by mood disturbance. This research aims at pointing out the specificities of olfactory perception in patients in order to identify the specific cerebral impairments involved in these disorders. METHODS: Olfactory sensitivity, detection, identification, self-evaluation of intensity and pleasantness have been recorded in a control group of healthy subjects (N = 58) and in three sample populations admitted to a Psychiatry Department: depressive patients (N = 49), anorectic patients (N = 17), and patients suffering from addiction to alcohol or drugs (N = 21). RESULTS: Depressive patients have a poor sensitivity, poor detection abilities but over-evaluate the pleasantness of odors. Anorectic patients have a high sensitivity, over-evaluate the intensities of the odors but under-evaluate their pleasantness. Alcoholic/drug addicted patients showed impairments in identification. LIMITATION: This study does not identify inter-individual differences in olfactory perception. CONCLUSION: The psychiatric diseases, here at hand although every one of them may be characterized by depressive components, show diverging impairments in olfactory perception. When variations in sensitivity are usually attributed to peripheral cues, impairments in emotional and cognitive aspects of olfaction are typically related to specific brain structures and processes which could be particularly involved in these diseases. PMID- 16376995 TI - Lymphopenia and spontaneous autorosette formation in SOD1 mouse model of ALS. AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by motoneuron degeneration. Increasing evidence suggests immune system involvement in ALS pathogenesis but information about peripheral blood characteristics has been lacking. We evaluated hematological and morphological parameters in peripheral blood of G93A SOD1 mice. A significant decrease in white blood cells was found at the end stage of disease. The lymphocyte reduction may suggest immunodeficiency in ALS. Spontaneously forming rosettes with autologous erythrocytes were noted in approximately 28% of lymphocytes in SOD1 mice. To our knowledge, this is the first study characterizing hematology and revealing autorosettes in the SOD1 mouse model of ALS at the terminal phase of disease. PMID- 16376996 TI - D3 dopamine receptor mRNA is elevated in T cells of schizophrenic patients whereas D4 dopamine receptor mRNA is reduced in CD4+ -T cells. AB - The expression of dopamine receptors was examined in purified human neutrophils, monocytes, B cells, natural killer cells and CD4+ - and CD8+ -T lymphocytes by RT PCR. In healthy subjects, D1 and D2 receptors were not expressed in leukocytes. Real Time PCR for dopamine receptors D3 and D4 disclosed that D3 receptors are expressed in T cells and natural killer cells and D4 receptors in CD4+ -T cells. The comparison of schizophrenic patients with sex- and age-matched controls revealed a significantly higher expression of D3 receptor mRNA in T cells of schizophrenic patients, whereas D4 receptor mRNA in CD4+ -T cells was downregulated. PMID- 16376997 TI - Targeting Rho and Rho-kinase in the treatment of cardiovascular disease. AB - The small GTPase Rho and its downstream effector Rho-kinase contribute to agonist induced vascular contraction via Ca2+ sensitization. Reasonably selective pharmacological inhibitors of these proteins have been developed and are now widely used experimentally to investigate the role of this signaling pathway in vascular function. Rho and Rho-kinase have attracted increasing clinical interest as a result of emerging evidence for their roles in the pathogenesis of several cardiovascular disorders, including hypertension, coronary and cerebral vasospasm, atherosclerosis and diabetes, and are now considered important future therapeutic targets. A major challenge lies in further developing selective inhibitors of this pathway beyond experimental use. Consideration should perhaps also be given to widening the application of existing clinical drugs now known to also interfere with Rho-Rho-kinase signaling. PMID- 16376998 TI - Novel class of pain drugs based on antagonism of NGF. AB - Nerve growth factor (NGF) was identified originally as a survival factor for sensory and sympathetic neurons in the developing nervous system. In adults, NGF is not required for survival but it has a crucial role in the generation of pain and hyperalgesia in several acute and chronic pain states. The expression of NGF is high in injured and inflamed tissues, and activation of the NGF receptor tyrosine kinase trkA on nociceptive neurons triggers and potentiates pain signalling by multiple mechanisms. Inhibition of NGF function and signalling blocks pain sensation as effectively as cyclooxygenase inhibitors and opiates in rodent models of pain. Several pharmaceutical companies have active drug discovery and development programs that are based on a variety of approaches to antagonise NGF, including NGF 'capture', blocking the binding of NGF to trkA and inhibiting trkA signalling. NGF antagonism is expected to be a highly effective therapeutic approach in many pain states, and to be free of the adverse effects of traditional analgesic drugs. PMID- 16376999 TI - Paradox of Ca2+ signaling, cell death and stroke. AB - Since the early 1960s it has been known that restoring extracellular Ca2+ following a period of low Ca2+ concentrations paradoxically causes an increase in intracellular Ca2+ levels that can lead to cell death. The mystery of this 'Ca2+ paradox' is made more intriguing by observations that lowering concentrations of extracellular Ca2+ and/or Mg2+ paradoxically enhances the entry of Ca2+ into hippocampal neurons. Until recently, the entry of Ca2+ through NMDA receptors was accepted as the major pathway leading to the excitotoxic, delayed cell death associated with the ischemic periods of stroke. Here, we discuss how several transient receptor potential (TRP) channels are likely to contribute to both the Ca2+ paradox and the delayed death of neurons following an ischemic stroke. PMID- 16377000 TI - Glial cell biology in Drosophila and vertebrates. AB - Glia are the most abundant cell type in the mammalian nervous system and they have vital roles in neural development, function and health. However our understanding of the biology of glia is in its infancy. How do glia develop and interact with neurons? How diverse are glial populations? What are the primary functions of glia in the mature nervous system? These questions can be addressed incisively in the Drosophila nervous system--this contains relatively few glia, which are well-defined histologically and amenable to powerful molecular-genetic analyses. Here, we highlight several developmental, morphological and functional similarities between Drosophila and vertebrate glia. The striking parallels that emerge from this comparison argue that invertebrate model organisms such as Drosophila have excellent potential to add to our understanding of fundamental aspects of glial biology. PMID- 16377001 TI - Treatment of intravaginal HSV-2 infection in mice: a comparison of CpG oligodeoxynucleotides and resiquimod (R-848). AB - The mammalian innate immune system recognizes pathogens via a series of pattern recognition receptors such as the toll-like receptors (TLR) that interact with pathogen-associated molecular patterns (PAMPs) and lead to the rapid activation of innate immune cells. In this study, we compared the efficacy of CpG ODN (a TLR9 agonist) and resiquimod (R-848; a TLR7/8 agonist) for topical immunoprophylaxis or immunotherapy of vaginal herpes simplex virus type 2 (HSV-2) infection in mice. Efficacy against HSV infection was observed with CpG ODN but less so with R-848, even after repeated administrations. Intravaginal (IVAG) administration of CpG ODN resulted in strong local but relatively weak systemic immune activation, as determined by levels of the chemokines IP-10, MIG and I-TAC in vaginal tissue and plasma, respectively. In contrast, IVAG administration of R 848 resulted in high levels of plasma IP-10, similar to those seen after parenteral administration, but overall, weaker or shorter-lived local immune responses than obtained with CpG ODN. These findings suggest that differences in biodistribution and sites of immune activation between CpG ODN and R-848 after IVAG delivery account for differences in efficacy, and demonstrate the need for local mucosal innate activation for protection against HSV-2. PMID- 16377002 TI - Microinjection of ritanserin into the dorsal hippocampal CA1 and dentate gyrus decrease nociceptive behavior in adult male rat. AB - Prenatal 5HT depletion causes a significant decrease in the level of nociceptive sensitivity during the second phase of the formalin test behavioral response. These experiments were designed to test whether blocking 5HT2A/2c receptors in the CA1 region of the hippocampus and dentate gyrus would decrease nociceptive behaviors induced by a peripheral noxious stimulus formalin as an animal model of unremitting human being. The 5HT2A/2c receptor antagonist ritanserin (2, 4 and 8 microg/0.5 microl) was injected into the CA1 area and dentate gyrus of behaving rats 5 min before subcutaneous injection of formalin irritant. Nociceptive behaviors in both phases of the formalin test were significantly decreased by ritanserin (4 and 8 microg/0.5 microl) and ritanserin had no effect at 2 microg/0.5 microl. These results support the hypothesis that the hippocampal formation may modify the processing of incoming nociceptive information and that 5HT2A/2c receptor-sensitive mechanisms in the hippocampus may play a role in nociception and/or the expression of related behaviors. PMID- 16377004 TI - Adrenergic modulation of the type 1 IP3 receptors in the rat heart. AB - Inositol 1,4,5-trisphosphate (IP3) receptors are calcium-releasing channels localized on the sarcoplasmic reticulum. IP3 receptors mediate the calcium mobilizing effect of a wide range of hormones, cytokines, and neurotransmitters and play an important role in variety of cell functions. The aim of this work was to study, how partial depletion of catecholamines affects the gene expression and protein levels of the type 1 IP3 receptors in rat heart. The type 1 IP3 receptor mRNA levels were studied in the left cardiac atrium and ventricle of rats treated with 6-hydroxydopamine (6-OHDA) in control and stressed conditions. The 6-OHDA produces anatomical and functional denervation resulting in decreased levels of noradrenaline and adrenaline. We also used corticoliberin (CRH) knockout mice, where secretion of adrenaline is significantly suppressed. Administration of 6 OHDA significantly decreases mRNA levels of the type 1 IP3 receptor in both, the left atrium and the left ventricle, while the gene expression of the sarcoplasmic reticular Ca2+-ATPase (SERCA 2) was unaffected. CRH knockout mice possess markedly lower levels of the type 1 IP3 receptor mRNA compared to wild-type mice in both, control and stressed conditions. These data point to the adrenergic modulation of the type 1 IP3 receptors in the rat hearts. PMID- 16377003 TI - The induction of NOS2 expression by the hybrid cecropin A-melittin antibiotic peptide CA(1-8)M(1-18) in the monocytic line RAW 264.7 is triggered by a temporary and reversible plasma membrane permeation. AB - There is an increasing awareness of immune cell modulation by antimicrobial peptides. While this process often requires specific receptors for the peptides involved, several reports point out to a receptor-independent process. The cecropin A-melittin hybrid peptide CA(1-8)M(1-18) (KWKLFKKIGIGAVLKVLTTGLPALIS amide) modifies gene expression in the macrophage line RAW 264.7 in the absence of any previous macrophage priming, suggesting a membrane permeation process. To further analyze the initial steps of this mechanism, we have studied the interaction of the peptide with these cells. Below 2 microM, CA(1-8)M(1-18) causes a concentration-dependent membrane depolarization partially reversible with time. At 2 microM, the accumulation of the SYTOX green vital dye is one half of that achieved with 0.05% Triton X-100. The binding level, as assessed by fluorescein-labeled CA(1-8)M(1-18), varies from 7.7+/-1.2 to 37.4+/-3.9 x 10(6) molecules/cell over a 0.5-4.0 microM concentration range. Electrophysiological experiments with 0.5 microM CA(1-8)M(1-18), a concentration that triggers maximal NOS2 expression and minimal toxicity, show a reversible current induction in the RAW 264.7 plasma membrane that is maintained as far as peptide is present. This activation of the macrophage involves the production of nitric oxide, a metabolite lethal for many pathogens that results from unspecific membrane permeation by antimicrobial peptides, and represents a new mode of action that may open new therapeutic possibilities for these compounds against intracellular pathogens. PMID- 16377005 TI - Magnetic resonance blood flow measurements in the follow-up of pediatric patients with aortic coarctation - a re-evaluation. AB - BACKGROUND: Previous studies have suggested the feasibility of a non-invasive quantification of vascular trans-stenotic pressure gradients (DeltaP) by phase contrast MR imaging (PC-MRI). Our purpose was to assess the value of MRI estimated pressure gradients as a screening tool for assessing hemodynamically significant (re-)coarctation of the aorta (CoA) in pediatric patients. METHODS: Forty-three patients (median age (range), 16 (5-25) years) with CoA (38 postoperative and 5 native) and clinically suspected hemodynamically significant stenosis underwent quantitative and semi-quantitative PC-MRI blood flow measurements and 3D MR-angiography, Doppler ultrasound (US) and conventional catheter angiography (CCA, n=20). Estimated DeltaP for each modality was correlated with percent stenosis. RESULTS: The percent stenosis correlated only moderately with DeltaP(MRI) (r=0.55, p<0.001) and DeltaP(CCA) (r=0.48, p<0.001). Only moderate correlations were observed between DeltaP(MRI) vs. DeltaP(CCA) (r=0.54, p=0.02) and vs. DeltaP(US) (r=0.40, p=0.01). In contrast, semi quantitative analysis of PC-MRI flow profiles predicted with good sensitivity (88%) and specificity (88%) who would be operated on. Thirteen patients met hemodynamic and percent stenosis criteria by CCA for surgical intervention. CONCLUSION: Measured pressure gradients using PC-MRI should be used cautiously when assessing patients for recoarctation of the aorta. The analysis of blood flow profiles by PC-MRI might be a promising alternative in assessing the hemodynamic significance of CoA. PMID- 16377006 TI - Detection of plaque of saphenous vein graft by multidetector row computed tomography and comparison with gray-scale/virtual histology intravascular ultrasound. PMID- 16377007 TI - Clinical significance of perfusion techniques utilising different physiological mechanisms to detect myocardial viability: a comparative study with myocardial contrast echocardiography and single photon emission computed tomography. AB - Myocardial uptake using (99m)Tc-sestamibi single photon emission computed tomography (SPECT) depends largely on myocardial microvascular volume. Myocardial contrast echocardiography (MCE) is a relatively new technique that detects not only microvascular volume but also blood flow. These differing mechanisms may affect the relative accuracies of MCE and SPECT for detecting myocardial viability (MV) early after acute myocardial infarction (AMI) and thrombolysis. Accordingly 56 patients underwent resting transthoracic echocardiography, low power MCE and SPECT 7+/-2 days following first AMI and thrombolysis. Contractile reserve (CR) was assessed 3 months following revascularization. The sensitivity and specificity of MCE and SPECT were 83% and 78% (p=ns) and 78% and 45% (p<0.01) respectively. MCE was the only multivariate predictor of global recovery of function and CR (OR=3.5, p=0.01). The different physiological mechanisms employed by MCE and SPECT translate into different relative accuracies for the detection of MV. PMID- 16377008 TI - Primary stenting of the left main coronary artery with anomalous origin from the right sinus of valsalva. AB - The presence of an aberrant origin of left main coronary artery from the right sinus of valsalva passing between the aorta ascendens and pulmonary trunk is one of the anomalies most frequently associated with malignant clinical events, and surgical treatment is recommended. We report of a 64-year-old patient with a highly increased risk for surgery due to severe chronic obstructive pulmonary disease. This case demonstrates that the use of a drug-eluting stent offers a suitable and valuable alternative. PMID- 16377009 TI - Self-blood letting in congestive cardiac failure. AB - A 33-year-old male with idiopathic restrictive cardiomyopathy had dilated and tortuous veins over both lower limbs and over the scrotum with multiple hyperpigmented scars. He was in the habit of puncturing the veins to let out blood during episodes of worsening of dyspnoea, thereby relieving dyspnoea by decreasing the preload. PMID- 16377010 TI - Clinical, biochemical and genetical resistance to clopidogrel in a patient with the recurrent coronary stent thrombosis--a case report and review of the literature. AB - The in-stent thrombosis is considered as a serious complication of percutaneous coronary intervention. The impaired response to the antiplatelet therapy may play an important role in this crucial problem. Recurrent thrombosis can be explained by genetic background. Because the phenomenon of clopidogrel resistance is associated with an increased risk of recurrent cardiovascular events, genetic variants of a platelet receptor P2Y(12) might be the risk factor. Although there is no test recommended to assess the response to the antiplatelet therapy, the presented case suggests platelet function analysis should be undertaken in patients with recurrent stent thrombosis. PMID- 16377011 TI - Patient outcomes when hospitals experience a surge in admissions. AB - In January 2004, a California law passed in 1999 became effective regulating minimum nurse to patient ratios in hospital units. This legislation was prompted by results from previous research showing higher adverse patient outcomes when hospital nurse/patient ratios are low. In this study, we use a census of hospital discharges in California over the 1996-2000 period to estimate the impact of hospital staff levels on adverse events by examining whether outcomes are correlated with the number of admissions in the hospital over the next 2 days. Because hospital staff levels are determined in advance, a large influx of patients to a hospital over the next 2 days will reduce the effective staff levels for patients admitted previously. We focus on the outcomes of patients admitted on Thursdays because hospital admissions are the most variable on Friday and Saturday and hospital staffs are lowest during the weekend. We find some evidence that large shocks to admissions on Friday and Saturday tend to reduce the length of stay and increase the chance of a subsequent readmission, but these coefficients are very small. We find quantitatively small and statistically insignificant effects of Friday and Saturday admission shocks on mortality rates of patients admitted on Thursdays. These results suggest that the portion of the California law designed to guarantee adequate staffing when the patient census increases unexpectedly should have little impact on patient outcomes. PMID- 16377012 TI - Removal of humic substances from soil DNA using aluminium sulfate. AB - Direct extraction of soil DNA has become essential for the study of soil microorganisms. Humic substances co-extracted during DNA retrieval is a big problem because it greatly inhibits the enzymes involved in manipulating DNA. Popular commercial kits available for soil DNA extraction are unable to overcome this problem. Here we report an effective protocol for the removal of humic substance from soil DNA. The protocol involves flocculation of the humic substance by excessive Al(3+), then removal of superfluous Al(3+) via pH adjustment and finally release of soil microbial DNA by SDS lysis. This technique is superior to that employed by the UltraClean Soil DNA Kit and can be applied to a wide variety of soils. PMID- 16377013 TI - Amplitude modulation of gamma band oscillations at alpha frequency produced by photic driving. AB - Gamma band response to visual stimulation in humans has been observed to have both burst and resonance properties. Amplitude modulation of gamma activity at low frequencies has been seen in rat hippocampus and modeled in a number of forms. Significant amplitude modulation (p=0.05) of 33 Hz gamma frequency activity at the frequency of an 8 1/3 Hz photic driving stimulus, which also produced strong alpha entrainment, was observed in 67% of the channels in 42 human subjects. Similar amplitude modulation was found at a range of frequencies from greater than 50 Hz to about 28 Hz. The peak of the gamma amplitude modulation curve trailed the peak of the alpha signal by 25 to 30 ms, corresponding to a phase difference of 150 degrees to 180 degrees. The phase consistency of the gamma signal, measured across comparable times of the alpha signal, was least at the minimum amplitude modulation, and largest at the maximum. Although there was no consistent overall relation between the gamma amplitude and alpha amplitude, peak gamma amplitude values were consistently higher during post-target-stimulus alpha suppression, which occurs about 300-750 ms subsequent to stimulus presentation, than they were at the time of maximum alpha activity during the immediate post-stimulus period. It is hypothesized that there is an interaction between the alpha and gamma generating systems, in which gamma triggers alpha activity and is subsequently inhibited by it, thus producing the observed amplitude modulation. The transition from dark to light of the photic driving stimulus begins a phase resetting process in the gamma system and a concomitant burst of gamma activity; this produces an activation in the alpha system, similar to that found in the P1-N1 response in evoked potential experiments, and a subsequent inhibition of gamma production. PMID- 16377014 TI - Roles of glial glutamate transporters in shaping EPSCs at the climbing fiber Purkinje cell synapses. AB - Glial glutamate transporters, GLAST and GLT-1, are co-localized in processes of Bergmann glia (BG) wrapping excitatory synapses on Purkinje cells (PCs). Although GLAST is expressed six-fold more abundantly than GLT-1, no change is detected in the kinetics of climbing fiber (CF)-mediated excitatory postsynaptic currents (CF EPSCs) in PCs in GLAST(-/-) mice compared to the wild-type mice (WT). Here we aimed to clarify the mechanism(s) underlying this unexpected finding using a selective GLT-1 blocker, dihydrokainate (DHK), and a novel antagonist of glial glutamate transporter, (2S,3S)-3-[3-(4-methoxybenzoylamino)benzyloxy]aspartate (PMB-TBOA). In the presence of cyclothiazide (CTZ), which attenuates the desensitization of AMPA receptors, DHK prolonged the decay time constant (tau(w)) of CF-EPSCs in WT, indicating that GLT-1 plays a partial role in the removal of glutamate. The application of 100 nM PMB-TBOA, which inhibited CF-mediated transporter currents in BG by approximately 80%, caused no change in tau(w) in WT in the absence of CTZ, whereas it prolonged tau(w) in the presence of CTZ. This prolonged value of tau(w) was similar to that in GLAST(-/-) mice in the presence of CTZ. These results indicate that glial glutamate transporters can apparently retain the fast decay kinetics of CF-EPSCs if a small proportion ( approximately 20%) of functional transporters is preserved. PMID- 16377015 TI - Internal initiation: IRES elements of picornaviruses and hepatitis c virus. AB - The scanning hypothesis provides an explanation for events preceding the first peptide bond formation during the translation of the vast majority of eukaryotic mRNAs. However, this hypothesis does not explain the translation of eukaryotic mRNAs lacking the cap structure required for scanning. The existence of a group of positive sense RNA viruses lacking cap structures (e.g. picornaviruses) indicates that host cells also contain a 5' cap-independent translation mechanism. This review discusses the translation mechanisms of atypical viral mRNAs such as picornaviruses and hepatitis c virus, and uses these mechanisms to propose a general theme for all translation, including that of both eukaryotic and prokaryotic mRNAs. PMID- 16377016 TI - Glutathione and thiolated chitosan inhibit multidrug resistance P-glycoprotein activity in excised small intestine. AB - The aim of the present study was to evaluate the influence of glutathione (GSH), the thiomer chitosan-4-thiobutylamidine (chitosan-TBA) and a combination of both compounds on P-glycoprotein (P-gp) activity. Permeation studies were performed with freshly excised guinea pig ileum mounted in Ussing chambers using the fluorescent dye rhodamine-123 (Rho-123) as P-gp substrate. Apparent permeability coefficients (Papp) as well as efflux ratios (secretory Papp/absorptive Papp) were calculated and compared with values gained from experiments with the well established P-gp inhibitors terfenadine and verapamil. In the presence of terfenadine, verapamil as well as GSH, the absorptive transport of Rho-123 across intestinal tissue increased, while the secretory decreased with efflux ratios around 1.0. Chitosan-TBA and especially chitosan-TBA/GSH not only enhanced absorption of Rho-123, but also reduced the basolateral to apical secretion of Rho-123 resulting in efflux ratios of 1.1, 0.8 and 0.5. The study indicates that chitosan-TBA/GSH is a potentially valuable tool for inhibiting the ATPase activity of P-gp in the intestine. PMID- 16377017 TI - One-step preparation of polyelectrolyte-coated PLGA microparticles and their functionalization with model ligands. AB - This work aimed at the development of a novel surfactant-free, one-step process for the concomitant formation of poly(lactide-co-glycolide) (PLGA) microparticles (MP) and surface coating with the polyelectrolyte chitosan, which is suitable for subsequent covalent conjugation of bioactive ligands. The technology is based on solvent extraction from an O/W-dispersion using a static micromixer. Surface coating occurred through interaction of the negatively charged, nascent PLGA MP with the polycationic chitosan, which was dissolved in the aqueous extraction fluid. Particles of 1-10 mum in diameter were produced with excellent reproducibility. The chitosan-coated PLGA MP were spherical and showed a smooth surface without pores, as demonstrated by scanning electron microscopy (SEM). The chitosan coatings were characterized by zeta potential measurements and X-ray photoelectron spectroscopy (XPS). The functional amino groups of chitosan were used to conjugate two model ligands to the coating, i.e. fluorescamine and NHS PEG-biotin. The presence of the conjugated ligands was revealed by confocal laser scanning microscopy (CLSM) and fluorescence activated cell sorting (FACS). Evidence for biotinylation was demonstrated through binding of fluorescently labelled streptavidin. The developed platform technology is straightforward and flexible. Future studies will focus on the design of microparticulate carriers with bioactive surfaces, e.g. as antigen delivery systems. PMID- 16377018 TI - Gastro-intestinal patch system for the delivery of erythropoietin. AB - The absorption of erythropoietin (EPO) from rat small intestine was studied using gastro-intestinal patches (GI-PS) in the presence of absorption enhancers. Surfactants such as a saturated polyglycolysed C8-C18 glyceride (Gelucire 44/14), PEG-8 capryl/caprylic acid glycerides (Labrasol), and polyoxyethylene hydrogenated castor oil derivative (HCO-60) were used as absorption enhancers at 143, 94 and 20 mg/kg, respectively. The absorption of EPO was studied by measuring serum EPO levels by an ELISA method after small intestinal administration of EPO-GI-PS preparation in rats at the EPO dose level of 100 IU/kg. Labrasol showed the highest absorption enhancing effect after intrajejunum administration with maximum serum EPO level of 84.1+/-11.4 mIU/ml while Gelucire 44/14 and HCO-60 showed 43.5+/-9.8 and 26.5+/-2.3 mIU/ml, respectively. The appropriate site for EPO absorption was also investigated. Jejunum was found to be the most efficient absorption site for the absorption of EPO from GI-PS. Using Labrasol as the absorption enhancer and jejunum as the absorption site, the effect of EPO dose on EPO absorption was studied by increasing the EPO dose from 50, to 100, 300 and 600 IU/kg. It was found that 100 IU/kg was the optimum dose with a serum EPO level of 84.1+/-11.4 mIU/ml while escalating doses showed decreases in serum EPO levels 48.3+/-5.6 for 300 IU/kg and 50.6+/-10.3 mIU/ml for 600 IU/kg. The percent bioavailability (BA) of EPO-GI-PS with Labrasol as absorption enhancer was 7.9 at 50 IU/kg, 12.1 at 100 IU/kg, 3.2 at 300 IU/kg and 1.2 at 600 IU/kg. Histological studies showed no adverse effect at the site of administration. PMID- 16377019 TI - Endosome trapping limits the efficiency of splicing correction by PNA-oligolysine conjugates. AB - Splicing correction by steric-blocking oligonucleotides (ON) might lead to important clinical applications but requires efficient delivery to cell nuclei. The conjugation of short oligolysine tails has been used to deliver a correcting peptide nucleic acid (PNA) sequence in a positive readout assay in which ON hybridization to the cryptic splice site is strictly required for the expression of a luciferase reporter gene. We have investigated the mechanism of cellular uptake and the efficiency of a (Lys)(8)-PNA-Lys construction in this model system. Cell uptake is temperature-dependent and leads to sequestration of the conjugate in cytoplasmic vesicles in keeping with an endocytic mechanism of internalization. Accordingly a significant and sequence-specific splicing correction is achieved only in the presence of endosome-disrupting agents as chloroquine or 0.5 M sucrose. These endosome-disrupting agents do not affect the activity of free PNA, and do not increase (Lys)(8)-PNA-Lys uptake. PMID- 16377020 TI - Association of the endothelial nitric oxide synthase (ecNOS) gene polymorphism with carotid atherosclerosis in type 2 diabetes. AB - This study was done to see whether 27-base pair repeats polymorphism in intron 4 of ecNOS gene is associated with carotid atherosclerosis in type 2 diabetic patients. The polymorphism was identified by polymerase chain reaction (PCR). Ultrasound parameters of carotid atherosclerosis were analyzed in relation to the genotype in 210 patients with type 2 diabetes. The ecNOS4a allele was detected in 34 (16.2%) of this study group. With the exception of the plaque count (P = 0.069), all other parameters obtained by ultrasound examination of carotid arteries were significantly correlated with presence of ecNOS4a allele (P < 0.05). As all the measured carotid parameters correlated well each other, we selected the total mean carotid IMT (intima-media thickness) value to be used for this analysis. In the multivariate analysis including several variables such as age, sex, hypertension, LDL cholesterol, waist-hip ratio, and fasting insulin, all determined to be significant by univariate analysis, ecNOS4a allele had a significant correlation with total mean IMT (P < 0.001). In conclusion, the ecNOS4a allele is associated with carotid atherosclerosis in type 2 diabetic patients in Korea. PMID- 16377021 TI - Concerns about methodology and use of 3,5,7-cholestatriene as a marker of oxidative stress Response to "Relationship between carotid atherosclerosis and erythrocyte membrane cholesterol oxidation products in type 2 diabetic patients". PMID- 16377022 TI - Serum CRP levels are equally elevated in newly diagnosed type 2 diabetes and impaired glucose tolerance and related to adiponectin levels and insulin sensitivity. AB - AIMS: To measure the serum highly sensitive C-reactive protein (hs-CRP) and adiponectin levels, assess insulin sensitivity index (SI) and acute insulin response (AIR) in normal control (NC) subjects, patients with impaired glucose tolerance (IGT) and newly diagnosed type 2 diabetes mellitus (DM), and further explore the possible correlation between hs-CRP and SI, AIR and adiponectin in IGT and newly diagnosed type 2 DM groups. METHODS: Age and sex matched 28 normal subjects, 31 patients with IGT, and 31 patients with newly diagnosed type 2 DM were included in the study. SI and AIR were assessed by the reduced sample number of Bergman's minimal model method with intravenous glucose tolerance test in subjects of each group. RESULTS: Compared with NC group, serum hs-CRP was significantly increased in IGT and type 2 DM groups (p < 0.001), although there was no significant difference between the latter groups. Hs-CRP was negatively correlated with high density lipoprotein cholesterol (HDL-C), SI and adiponectin levels (p < 0.05 to p < 0.001), and positively correlated with systolic blood pressure (SBP), fasting plasma glucose (FPG), BMI, waist-to-hip ratio (WHR), postprandial 2h plasma glucose (2hPG), fasting serum insulin (FINS) and postprandial serum insulin (PSI) in IGT and newly diagnosed type 2 DM groups (p < 0.05 to p < 0.001). In general multivariate regression, only adiponectin was the significantly independent determinant for serum hs-CRP (regression coefficient 1.380; 95% CI -2.062 to 0.698, p < 0.001); meanwhile, TG, SI, hs-CRP, FINS, 2hPG and WHR were significantly independent determinants for serum adiponectin concentration (p < 0.05 to p < 0.001). CONCLUSIONS: Elevated serum hs-CRP may play a role in the development of insulin resistance syndrome and type 2 diabetes. This elevation is accompanied by the opposite changes of adiponectin. PMID- 16377023 TI - Leadership and priority setting: the perspective of hospital CEOs. AB - The role of leadership in health care priority setting remains largely unexplored. While the management leadership literature has grown rapidly, the growing literature on priority setting in health care has looked in other directions to improve priority setting practices--to health economics and ethical approaches. Consequently, potential for improvement in hospital priority setting practices may be overlooked. A qualitative study involving interviews with 46 Ontario hospital CEOs was done to describe the role of leadership in priority setting through the perspective of hospital leaders. For the first time, we report a framework of leadership domains including vision, alignment, relationships, values and process to facilitate priority setting practices in health services' organizations. We believe this fledgling framework forms the basis for the sharing of good leadership practices for health reform. It also provides a leadership guide for decision makers to improve the quality of their leadership, and in so doing, we believe, the fairness of their priority setting. PMID- 16377024 TI - Sex is a threshold dichotomy mimicking a single gene effect. AB - The discovery of SRY ended the search for the male-determining gene, but the much sought after molecular pathway that connects the gene to its phenotype remains elusive. It is timely, therefore, to consider an alternative route of inheritance that can simulate mendelian segregation. The term "threshold dichotomy" is applied to contrasting phenotypes that have their origin in multiple genes with quantitative effects and are divided by a physiological threshold. I suggest that this mechanism provides a plausible basis for male development in mammals, which is known to depend on enhanced rates of cell proliferation in the developing embryo and can be expected to require increased levels of cellular energy. The system is affected by the genetic and environmental background. I also propose that the Y chromosome minimizes the effects of these variables on XY embryos so that, barring rare exceptions, the inheritance of sex resembles a single gene effect. PMID- 16377025 TI - Inferential literacy for experimental high-throughput biology. AB - Many biologists believe that data analysis expertise lags behind the capacity for producing high-throughput data. One view within the bioinformatics community is that biological scientists need to develop algorithmic skills to meet the demands of the new technologies. In this article, we argue that the broader concept of inferential literacy, which includes understanding of data characteristics, experimental design and statistical analysis, in addition to computation, more adequately encompasses what is needed for efficient progress in high-throughput biology. PMID- 16377026 TI - Assessment of biogeochemical natural attenuation and treatment of chlorinated solvents, Altus Air Force Base, Altus, Oklahoma. AB - Biogeochemical reductive dechlorination (BiRD) is a newly recognized method for the remediation or natural attenuation of chlorinated solvents. Chlorinated solvents are rapidly treated by abiotic reaction with reduced mineral iron sulfides. Iron sulfides are formed by naturally occurring sulfate-reducing bacteria when sufficient SO(4)(2-) and organic carbon are present or supplied to sediments containing mineral iron. An example of site characterization focusing on BiRD is presented focusing on mineral phases. Methods demonstrated here may be employed at other sites to evaluate naturally occurring BiRD or to evaluate an engineered BiRD remediation. A field investigation was performed at a TCE contaminated site at Altus AFB with naturally high concentrations of SO(4)(2-) and Fe(III) minerals and where an accidental fuel spill provided organic carbon. In the area of this fuel spill significant mineral iron sulfides were found, sulfate was almost completely removed, and TCE was absent. Only small amounts of daughter products were found, further indicating that the BiRD pathway was operative. Mass balance data indicates all of the remaining TCE (182 kg) could be treated by the remaining FeS (66.5 kg) in the upper aquifer; however, the FeS was not co-located with TCE to enable complete reaction. Laboratory microcosm tests with FeS amended and FeS-rich sediment from Altus AFB also suggest that BiRD is capable of destroying TCE. The results suggest that an engineered BiRD treatment is possible for this site. PMID- 16377027 TI - Numerical simulation of reactive processes in an experiment with partially saturated bentonite. AB - Bentonites are preferred materials for use as engineered barriers for high-level nuclear waste repositories. Simulation of geochemical processes in bentonite is therefore important for long-term safety assessment of those repositories. In this work, the porewater chemistry of a bentonite sample subject to simultaneous heating and hydration, as studied by Cuevas et al. [Cuevas, J., Villar, M., Fernandez, A., Gomez, P., Martin, P., 1997. Porewaters extracted from compacted bentonite subjected to simultaneous heating and hydration. Applied Geochemistry 12, 473-481.], was assessed with a non-isothermal reactive transport model by coupling the geochemical software PHREEQC2 with the object-oriented FEM simulator GeoSys/RockFlow. Reactive transport modelling includes heat transport, two-phase flow, multicomponent transport and geochemical reactions in the liquid phase, i.e. ion exchange, mineral dissolution/precipitation and equilibrium reactions. Simulations show that the easily soluble minerals in bentonite determine the porewater chemistry. Temperature affects both two-phase flow and geochemical reactions. Porosity change due to dissolution/precipitation is low during the experiment. However, changes of the effective porosity caused by bentonite swelling can be very large. The simulated results agree well with the experimental data. PMID- 16377028 TI - Methicillin-resistant Staphylococcus aureus: awareness and perceptions. AB - The purpose of this study was to investigate levels of awareness and the perceived risk of contracting methicillin-resistant Staphylococcus aureus (MRSA) amongst patients/visitors and employees of the UK National Health Service (NHS). Differences between the two cohorts were also investigated, particularly sources of information about MRSA. In March 2005, a piloted questionnaire was circulated to 50 patients/visitors and 100 NHS employees (25 doctors, 25 nurses, 25 domestics and 25 porters). No information was returned from porters, but the overall response rate was 67%. There was a high level of awareness of MRSA among both patients/visitors (94%) and NHS employees (100%). General media was the most common source of information for patients/visitors (68%) compared with 24% of NHS employees (P < 0.01). Perceived risk of contracting MRSA was very similar among patients/visitors and NHS employees (34% vs 35%, P > 0.10). Fifty-two percent of doctors felt that they were at risk compared with 13% of domestic employees. In conclusion, this study showed a high level of awareness among the general public and healthcare workers alike. PMID- 16377029 TI - Surveillance for mupirocin resistance following introduction of routine peri operative prophylaxis with nasal mupirocin. AB - The authors have previously described the successful use of a five-day peri operative prophylaxis regimen using nasal mupirocin and topical triclosan (PPNMTT) to prevent methicillin-resistant Staphylococcus aureus (MRSA) infection. The present article describes the results of repeated point-prevalence surveillance for four years to determine whether mupirocin resistance has emerged in surgical units using empirical, short-term, peri-operative prophylaxis with nasal mupirocin. Before starting PPNMTT and every six months thereafter for four years, point-prevalence surveillance was performed for nasal S. aureus carriage in all patients on five orthopaedic surgery wards, one vascular surgery ward and one elderly medicine control ward. S. aureus screening and clinical isolates (surgical patients) were undertaken for low- [minimum inhibitory concentration (MIC) 8-128 mg/L] and high-level (MIC > 128 mg/L) mupirocin resistance. All isolates were phage typed to determine whether there was evidence of the spread of clonal mupirocin-resistant strains. Of 593, 139 and 206 nasal screening swabs (taken after the regimen had started) from orthopaedic, vascular and control patients, 28%, 24% and 48% (orthopaedic/vascular surgery vs elderly medicine, P < 0.001) yielded S. aureus isolates, respectively, and 12%, 11% and 30% (P < 0.001) were MRSA positive, respectively. Of the S. aureus nasal screen isolates from orthopaedic/vascular surgery and control patients, 5% and 4%, respectively, were low-level mupirocin resistant (P > 0.1). Of 286 (orthopaedic/vascular surgery) and 68 (elderly medicine) clinical S. aureus isolates obtained after the regimen had started, 7% and 9% (P > 0.1), respectively, were low-level mupirocin resistant. No high-level mupirocin-resistant isolates were isolated from mupirocin (orthopaedic/vascular surgery) or elderly medicine control ward patients. There was no trend towards increasing prevalence of low-level mupirocin resistance during the four-year study period. The results of phage typing did not support the clonal spread of resistant strains. Long-term follow-up confirmed the efficacy of PPNMTT in reducing the prevalence of nasal carriage of S. aureus and MRSA in orthopaedic and vascular surgery patients. Despite four years of use of PPNMTT, there was no evidence of sustained emergence or spread of mupirocin resistance. PMID- 16377030 TI - Infectious complications of implantable ports and Hickman catheters in paediatric haematology-oncology patients. AB - The aim of this study was to define and compare the infectious and non-infectious complications associated with Hickman catheters and implantable ports in children. The study was conducted over a three-year period in the Department of Haematology-Oncology at the Schneider Children's Medical Center of Israel. All patients who required a central venous catheter (CVC) were included in the study. For each episode of catheter-associated bloodstream infection, demographic, clinical and microbiology data were recorded. During the study period, 419 tunnelled CVCs (246 implantable ports and 173 Hickman) were inserted in 281 patients. Compared with implantable ports, Hickman catheters were associated with a significantly higher rate of bloodstream infections (4.656 vs 1.451 episodes per 1000 catheter-days), shorter time to first infection (52.31 vs 108.82 days, P < 0.001), shorter duration of catheterization (140.75 vs 277.28 days, P < 0.001), and higher rate of removal because of mechanical complications (P < 0.005). Gram positive bacterial infections were more prevalent in the implantable port group (63.6% vs 41.6%), whereas Gram-negative rods, polymicrobial infections and mycobacterial infections were more prevalent in the Hickman group (31.4% vs 50.9%, 17% vs 36% and 0% vs 4.4%, respectively; P < 0.05 for all). Haematopoietic stem cell transplantation was identified as an independent risk factor for infection [odds ratio (OR) -1.68, P = 0.005] and for catheter removal due to complications (OR -2.0, P < 0.001). Implantable ports may be considered the preferred device for most paediatric oncology and stem cell transplantation patients. PMID- 16377031 TI - Structural analysis of peptides that interact with Newcastle disease virus. AB - A peptide with the sequence CTLTTKLYC has previously been identified to inhibit the propagation of Newcastle disease virus (NDV) in embryonated chicken eggs and tissue culture. NDV has been classified into two main groups: the velogenic group, and mesogenic with lentogenic strains as the other group based on its dissociation constants. In this study the peptide, CTLTTKLYC, displayed on the pIII protein of a filamentous M13 phage was synthesized and mutated in order to identify the amino acid residues involved in the interactions with NDV. Mutations of C1 and K6 to A1 and A6 did not affect the binding significantly, but substitution of Y8 with A8 dramatically reduced the interaction. This suggests that Y8 plays an important role in the peptide-virus interaction. The three dimensional structure of the peptide was determined using circular dichroism (CD), nuclear magnetic resonance (NMR), and molecular modeling. The peptide exhibited two possible conformers. One that consists of consecutive beta-turns around T2-L3-T4-T5 and K6-L7-Y8-C9. The other conformer exhibited a beta-hairpin bend type of structure with a bend around L3-T4-T5-K6. PMID- 16377032 TI - FMRFamide related peptide ligands activate the Caenorhabditis elegans orphan GPCR Y59H11AL.1. AB - G-protein coupled receptors (GPCRs) are ancient molecules that can sense environmental and physiological signals. Currently, the majority of the predicted Caenorhabditis elegans GPCRs are orphan. Here, we describe the characterization of such an orphan C. elegans GPCR, which is categorized in the tachykinin-like group of receptors. Since the C. elegans genome predicts only one tachykinin-like peptide (SFDRMGGTEFGLM), which could not activate the receptor, we hypothesized that one or some of the numerous FMRFamide related peptides (FaRPs) could be the cognate ligands for this receptor. This hypothesis was based on the suggestion that RFamides may be ancestral neuropeptides, from which a lot of the amidated neuropeptides, including tachykinins, derived. Indeed, we found that the orphan receptor encoded by the Y59H11AL.1 gene is activated by several C. elegans neuropeptides, including SPMERSAMVRFamide. These peptides activate the receptor in a concentration-dependent way. PMID- 16377033 TI - Increased NGF proforms in aged sympathetic neurons and their targets. AB - Target-derived neurotrophins such as nerve growth factor (NGF) and neurotrophin-3 (NT-3) regulate sympathetic neuron survival. Here, NGF and NT-3 protein and transcript were examined in sympathetic neurons and targets in order to determine their role in age-related neuronal atrophy. One obvious alteration was a dramatic increase (up to 50-fold) in NGF protein forms, corresponding to proNGF-B, in the superior cervical ganglion (SCG) and targets where sympathetic innervation shows atrophy. In the iris, where sympathetic innervation is protected into old age, proNGF-B was decreased. Alterations in NGF transcript paralleled changes in NGF protein, albeit to a lesser degree. Though significantly increased in aged SCG, NT-3 protein, found primarily as the 'mature' form, showed only minor changes in most tissues, though NT-3 mRNA generally was decreased. In contrast, both NT-3 transcript and NT-3 precursors were increased in iris. The dramatic increases in proNGF, together with minimal changes in NT-3, suggest that alterations in NGF regulation may contribute to the loss of sympathetic innervation observed in many aged peripheral targets. PMID- 16377034 TI - Synthesis, structure and anticancer activity of novel 2,4-diamino-1,3,5-triazine derivatives. AB - A series of 2-(4,6-diamino-1,3,5-triazin-2-yl)-2-{[4-(dimethylamino) phenyl]imino}acetonitriles 19-27 have been synthesized by the reaction of 2-(4 amino-6-alkylamino-1,3,5-triazin-2-yl)acetonitriles 10-15 with p nitrosodimethylaniline. Unexpectedly, a similar reaction of acetonitriles 10, 14, 15, 17 and 18 with nitrosobenzene led to the formation of 4,6-diamino-N-phenyl 1,3,5-triazine-2-carboxamides 28-32. The in vitro antitumor activity of the compounds obtained has been tested and 2-[4-Amino-6-(4-phenylpiperazin-1-yl) 1,3,5-triazin-2-yl]-2{[4-(dimethylamino)phenyl]imino}acetonitrile (19) having remarkable activity against melanoma MALME-3 M cell line (GI(50)=3.3 x 10(-8) M, TGI=1.1 x 10(-6) M) is a leading candidate for further development. PMID- 16377035 TI - Synthesis and in vitro selective anti-Helicobacter pylori activity of N substituted-2-oxo-2H-1-benzopyran-3-carboxamides. AB - In order to develop new anti-Helicobacter pylori agents, five new and three already known N-substituted-2-oxo-2H-1-benzopyran-3-carboxamides (coumarin-3 carboxamides) were prepared and evaluated for their antibacterial activity. All synthesized compounds showed little or no activity against different species of Gram-positive and Gram-negative bacteria of clinical relevance and against various strains of pathogenic fungi. Among the prepared compounds those with a 4 acyl-phenyl group showed the best activity against H. pylori metronidazole resistant strains in the 0.25-1 microg/ml MIC range, indicating the presence of an acyl function as an important feature for activity. PMID- 16377036 TI - Immunogenicity and protective efficacy of Escherichia coli expressed Plasmodium falciparum merozoite surface protein-1(42) using human compatible adjuvants. AB - The C-terminal 42-kDa fragment of the merozoite surface protein-1 of Plasmodium falciparum (PfMSP-1(42)) was expressed as a recombinant protein in Escherichia coli and purified to near homogeneity. We tested the immunogenicity of recombinant PfMSP-1(42) in three clinically acceptable adjuvants (Montanide ISA 720, alum and MF59) in mice and in rabbits. High antibody responses were obtained with two adjuvant formulations with IgGl being the predominant immunoglobulin isotype. Significant T-cell proliferation responses were also observed. Competitive enzyme linked immunosorbant assay (ELISA) showed the presence of both invasion and processing inhibitory antibodies in sera obtained from the immunized rabbits. Passive immunizations of mice with anti-PfMSP-1(42) IgG purified from the rabbit-sera were found to be protective against a parasite challenge with P. berghei/P. falciparum chimeric line (Pb-PfM19) that expresses Plasmodium falciparum MSP-1(19). These findings may be useful for the development of a malaria vaccine based on Plasmodium falciparum MSP-1(42). PMID- 16377037 TI - Respiratory immunity is an important component of protection elicited by subunit vaccination against pneumonic plague. AB - Mice were vaccinated with a recombinant fusion protein, rF1-V, by an intramuscular prime followed by an intranasal boost, to evaluate protection against pneumonic plague. Forty-two days after the intranasal boost, the mice were challenged by aerosol exposure to Yersinia pestis. Survival after exposure depended upon the dose of rF1-V given i.n. with > or = 80% survival in the highest dose groups. Pulmonary and serum antibody titers to V were the best predictors of outcome. For vaccinated mice that succumbed to the infection, death was delayed by 1-2 days compared to sham-inoculated controls. Weight loss early after exposure correlated with outcome. Pathology studies indicated a severe, necrotizing bronchopneumonia in vaccinated mice that succumbed to the infection, compatible with a prolonged disease course, while the lungs of sham-inoculated mice had only mild pneumonia, which is compatible with a more rapid disease course. Immunity in the respiratory tract appears to be critical for protection against primary pneumonia caused by Y. pestis. PMID- 16377038 TI - Parameterization of a dynamic specific activity model of 14C transfer from surface water-to-humans. AB - Carbon-14 is a particularly interesting radionuclide from the perspective of dose estimation because it mixes readily with stable CO2, and hence enters the food chain as fundamental biomolecules. A model was developed for the situation of 14C releases to surface waters, where there are distinct changes in the water 14C activity concentrations throughout the year. The model computes the specific activity in water, phytoplankton, fish, crops, meat, milk and air, following a typical irrigation-based food-chain scenario. This paper describes the derivation of the required 14C-specific parameter values. Many of the key parameters are not commonly measured, at least not in the context of dose assessment. Thus, inference from other sources of data was required, and this is the scientific contribution described in this paper. The best estimates and appropriate measures of statistical dispersion are provided. This required consideration of both the temporal and spatial averaging domains to ensure they were correct for parameters as defined in the model. The model coupled with these parameter values represents several new developments for modelling 14C transfers. PMID- 16377039 TI - Choosing an alpha radiation weighting factor for doses to non-human biota. AB - The risk to non-human biota from exposure to ionizing radiation is of current international interest. In calculating radiation doses to humans, it is common to multiply the absorbed dose by a factor to account for the relative biological effectiveness (RBE) of the radiation type. However, there is no international consensus on the appropriate value of such a factor for weighting doses to non human biota. This paper summarizes our review of the literature on experimentally determined RBEs for internally deposited alpha-emitting radionuclides. The relevancy of each experimental result in selecting a radiation weighting factor for doses from alpha particles in biota was judged on the basis of criteria established a priori. We recommend a nominal alpha radiation weighting factor of 5 for population-relevant deterministic and stochastic endpoints, but to reflect the limitations in the experimental data, uncertainty ranges of 1-10 and 1-20 were selected for population-relevant deterministic and stochastic endpoints, respectively. PMID- 16377040 TI - A congenital teratoma with a cleft palate: report of a case. AB - Teratomas are benign tumors containing cells from ectodermal, mesodermal and endodermal layers. They occur in about 1 in every 4000 births and most commonly in the sacrococcygeal region, followed by the ovaries. Congenital epignathus teratomas are rare embryological neoplasms localised in the region of head and neck. An epignathus is found in approximately 1:35,000 to 1:200,000 live births. This accounts for 2-9% of all teratomas. Size and location of the neoplasm in the oronasopharynx is variable. Teratomas are partly undiagnosed at the time of birth. They may exist with an intracranial extension or as small polyps. Large epignathi can lead to difficult management during and after birth. The case of a newborn girl with a combination of an epignathus and a cleft palate is described. The epignathus presented as a huge mass extending out of the mouth of the infant girl. On the day of birth debulking of the extraoral portion of the tumor, followed by intraoral exstirpation, was performed. The results of the histologic examination indicated a congenital epignathus. Six months later a recurrence was found. PMID- 16377041 TI - Physicochemical and biological characterisation of different dredged sediment deposit sites in France. AB - The aim of this work is to determine sediment properties, metal contents and transfers of Cd and Zn from dredged sediments to plants. To this end 10 deposit sites with different contexts were visited in France. The main agronomic characteristics and metal contents for surface soil layers were measured, the plant species present at the sites, such as Brassicaceae and Fabaceae, were listed, and the distribution of their root systems described. Soil characteristics such as available P (Olsen) varied between sites, with values ranging from 0.01 to 0.49 g kg(-1). Total contents and enrichment factors were studied, highlighting metal contamination in most of the sites. Despite carrying out principal component analyses, it was not possible to group deposits by age or geographical localisation. However, deposits could be distinguished as a function of proximity of industrial facilities, sediment grain size and carbonate content. Associations between metals were also highlighted: (1) Cd, Pb and Zn, and (2) Al, Cr, Cu and Fe. Consequently, we propose classifying them as technogenic anthrosols. PMID- 16377043 TI - Acid sulphate soil disturbance and metals in groundwater: implications for human exposure through home grown produce. AB - A significant emerging environmental problem is the disturbance and oxidation of soils with high levels of iron sulphide minerals resulting in acidification and causing the mobilization of metals into groundwater. This process is occurring in many parts of the world. In Western Australia, impacted groundwater is extracted by residents for domestic use. We sought to establish domestic use patterns of bore water and the concentration of metals. Sixty-seven domestic bore water samples clearly indicated oxidation of sulphidic materials with heavy metal concentrations ranging for aluminium (male for CYP2A4, CYP2B9, CYP2B10, and CYP3A41, and male>female for CYP2D9. In mice neonatally treated with monosodium L glutamate to produce GH-deficiency, GH was found to be an essential factor with GH archetype as a determinant in the regulatory mechanism of hepatic CYP2D9 and CYP3A41 expression, and GH was shown to be a repressive factor for the constitutive expression in females. Implantation with micro-osmotic pump containing GH (to yield a constant release of GH to mimic the plasma GH profile in females) to male mice increased CYP2A4, CYP2B9, CYP2B10, and CYP3A41, but decreased CYP2D9, expression to female levels, while conversely, twice-daily administration of GH (to produce the so-called male pattern of plasma GH levels) to female mice resulted in the repression of female-specific, CYP2B9 and CYP3A41, as well as female-predominant, CYP2A4 and CYP2B10, expression, and induction of male-specific CYP2D9 expression. Thus, the sex-dependent plasma GH profile (referred to hereafter as the GH archetype) was a decisive factor for the expression of sex-specific P450 genes in adult mouse liver. On the other hand, the regulation of CYP1A2, CYP2C29, and CYP3A11 expression was either sex independent or GH archetype-independent, considering the comparable levels between sexes of the constitutive expression and GH-inducible expression of these isoforms. Moreover, the observations suggested for the first time that the expression of CYP2B9 and CYP2A4 was not entirely GH-independent, but rather involved an imprinting GH-related factor that participated in the regulatory mechanism of P450 expression in females. PMID- 16377054 TI - Geranylgeranylacetone protects against acetaminophen-induced hepatotoxicity by inducing heat shock protein 70. AB - Geranylgeranylacetone (GGA), an anti-ulcer drug, has been reported to induce heat shock protein (HSP) 70 in several animal organs. The present study was performed to determine whether GGA protects mouse liver against acetaminophen (APAP) induced injury and whether it has potential as a therapeutic agent for APAP overdose. Hepatic damage was induced by single oral administration of APAP (500 mg/kg). GGA at 400 mg/kg was given orally 4 or 8h before, or 0.5h after APAP administration. Treatment of mice with GGA 4h before or 0.5h after APAP administration suppressed increases in transaminase activities and ammonia content in blood as well as hepatic necrosis. Such GGA treatment significantly increased hepatic HSP70 accumulation after APAP administration. Furthermore, GGA inhibited increases in hepatic lipid peroxide content and hepatic myeloperoxidase activity after APAP administration. In contrast, GGA neither inhibited hepatic cytochrome P450 2E1 activity nor suppressed hepatic glutathione depletion after APAP administration. The protective effect of GGA treatment 4h before APAP on hepatotoxicity induced by APAP was completely inhibited with quercetin, known as an HSP inhibitor. In conclusion, GGA has been identified as a new antidote to APAP injury, acting by induction of HSP70. The potential of GGA as a therapeutic tool is strongly supported by its ability to inhibit hepatic injury even when administered after ingestion of APAP. PMID- 16377055 TI - Plant metallothionein domains: functional insight into physiological metal binding and protein folding. AB - Plant metallothioneins (MTs) differ from animal MTs by a peculiar sequence organization consisting of two short cysteine-rich terminal domains linked by a long cysteine-devoid spacer. The role of the plant MT domains in the protein structure and functionality is largely unknown. Here, we investigate the separate domain contribution to the in vivo binding of Zn and Cu and to confer metal tolerance to CUP1-null yeast cells of a plant type 2 MT (QsMT). For this purpose, we obtained three recombinant peptides that, respectively, correspond to the single N-terminal (N25) and C-terminal (C18) cysteine-rich domains of QsMT, and a chimera in which the spacer is replaced with a four-glycine bridge (N25-C18). The metal-peptide preparations recovered from Zn- or Cu-enriched cultures were characterized by ESI-MS, ICP-OES and CD and UV-vis spectroscopy and data compared to full length QsMT. Results are consistent with QsMT giving rise to homometallic Zn- or Cu-MT complexes according to a hairpin model in which the two Cys-rich domains interact to form a cluster. In this model the spacer region does not contribute to the metal coordination. However, our data from Zn-QsMT (but not from Cu-QsMT) support a fold of the spacer involving some interaction with the metal core. On the other hand, results from functional complementation assays in endogenous MT-defective yeast cells suggest that the spacer region may play a role in Cu-QsMT stability or subcellular localization. As a whole, our results provide the first insight into the structure/function relationship of plant MTs using the analysis of the separate domain abilities to bind physiological metals. PMID- 16377056 TI - Gamma-irradiated RecD overproducers become permanent recB-/C- phenocopies for extrachromosomal DNA processing due to prolonged titration of RecBCD enzyme on damaged Escherichia coli chromosome. AB - The RecBCD enzyme of Escherichia coli consists of three subunits RecB, RecC and RecD. RecBCD enzyme activities are regulated by its interaction with recombination hotspot Chi. Biochemical and genetic evidence suggest that interaction with Chi affects RecD subunit, and that RecD polypeptide overproduction antagonizes this interaction, suggesting that intact RecD replaces a Chi-modified one. We used bacteria with fragmented chromosomes due to double strand breaks inflicted by UV and gamma-irradiation to explore in which way increased concentrations of RecBCD's individual subunits affect DNA metabolism. We confirmed that RecD overproduction alters RecBCD-dependent DNA repair and degradation in E. coli. Also, we found that RecB and RecC overproduction did not affect these processes. To determine the basis for the effects of RecD polypeptide overproduction, we monitored activities of RecBCD enzyme on gamma damaged chromosomal DNA and, in parallel, on lambda and T4 2 phage DNA duplexes provided at intervals. We found that gamma-irradiated wild-type bacteria became transient, and RecD overproducers permanent recB(-)/C(-) phenocopies for processing phage DNA that is provided in parallel. Since this inability of irradiated bacteria to process extrachromosomal DNA substrates coincided in both cases with ongoing degradation of chromosomal DNA, which lasted much longer in RecD overproducers, we were led to conclude that the RecB(-)/C(-) phenotype is acquired as a consequence of RecBCD enzyme titration on damaged chromosomal DNA. This conclusion was corroborated by our observation that no inhibition of RecBCD activity occurs in gamma-irradiated RecBCD overproducers. Together, these results strongly indicate that RecD overproduction prevents dissociation of RecBCD enzyme from DNA substrate and thus increases its processivity. PMID- 16377057 TI - Wolff-Parkinson-White syndrome mimicking acute inferolateral myocardial ischaemia in pneumonia. PMID- 16377058 TI - Quality of BLS decreases with increasing resuscitation complexity. AB - OBJECTIVE: Multiple procedures performed in parallel may cause each procedure to be performed less effectively than if performed in isolation. BLS performed by prehospital providers potentially includes artificial ventilations, chest compressions, and application of an automated external defibrillator (AED). This study examines the effectiveness of artificial ventilation and chest compressions both with and without an AED. METHODS: Thirty-six prehospital providers participated in a prospective observational study. Tested in pairs (n=18), subjects randomly completed three, 6-min scenarios [apneic patient with a pulse (VENT), a pulseless patient (CPR), and a pulseless patient with an AED available (CPR+AED)]. A full-torso manikin capable of generating a carotid pulse was connected to a computer to record number of ventilations, tidal volume, flow rate, number of compressions, and compression depth. Data were analyzed by t test, ANOVA, and Mann-Whitney U-test. RESULTS: Artificial ventilation performed in isolation provided more correct ventilations than during CPR or CPR+AED (25.7%, 14.2%, 13.7%, p=0.02). Fewer ventilations were delivered during CPR and CPR+AED (p=0.03). More compressions were delivered with CPR alone vs. CPR+AED (51.9, 35.7 min(-1), p=0.00). More correct compressions were delivered during CPR alone vs. CPR+AED (p=0.05). CONCLUSIONS: Both the quality and quantity of BLS decreases as the number of procedures performed simultaneously increases. Further decrements might occur when ALS skills enter into resuscitation. These results suggest a need to automate and/or prompt the performance of BLS to optimize resuscitation. PMID- 16377059 TI - The simple perfection of quantum correlation in human vision. AB - A theory is presented that specifies the amount of light that is needed for the perception of any stimulus that is defined in space, time and color. For detection and discrimination mechanistic neural elements with deterministic procedures exist. Twin pairs of red and green cones are ordered in three sets along clockwise and counter clockwise revolving spirals and along circles around the center of the fovea. In the rod-free fovea the red pairs are ordered along the spirals and the green along the circles. Each cone is accompanied by- dependent on retinal eccentricity--up to 100 satellite rods. For the retinal signal processing such a receptor group constitutes a space-quantum in analogy with time-quanta of about 0.04 s. In the peripheral retina the red and green twin pairs of space-quanta are roughly ordered along and at random distributed over the spirals and circles. Over each time-quantum, the cone and rods of a space quantum sum their responses in a common nerve circuit of the luminosity channel. The summation's results from twin pairs of the same set of space-quanta are correlated by two-fold spatio-temporal coincidence mechanisms in the retina. Their outcome signals the perception of light, movement and edge. In the fused binocular visual field the movement and edge signals of the three sets from both eyes perfectly join vectorially together, provided the responding pairs of space quanta are binocularly in perfect register as they normally are. The receptor's Weber gain control makes the receptor an all-or-none-system. The space-quantum's De Vries gain control makes its sensitivity equal to the average of the poisson fluctuations in quantum absorption per time-quantum. The controls are based on, respectively, arithmetically feed forward and backward inhibitive nerve mechanisms. The thermal noise of the photo-pigment resets the controls. The response to the second quantum absorption in a time-quantum in the individual rod, red or green cone has accession to the white, red or green nerve color circuit, respectively, and produces there a corresponding color signal. Already a single absorption in a blue cone is for a blue signal. In the retina, for the generation of yellow signals, the color circuits of individual red and green cones of each mixed entwined triple of red and green twin pairs of space-quanta are cross-connected through a nerve opponent color circuit. In the lateral geniculate nucleus in groups of seven neighboring triples, through two nerve opponent color circuits that are common for the two eyes together, the red and green signals as well as the yellow and blue mutually annihilate each other's color. White signals remain. In anomalous trichromacy, the space-quanta of some pairs have different cones or in one of them the cone is missing. In dichromacy, all pairs have different cones or one type of cones is missing. For perceptive resolution the periodic scanning of the retinal image by the eye tremor in synchrony with the time-quanta, overrules the limit of optical resolution as set by diffraction in the eye optics. Dependent on pupil diameter the scanning contributes up to a factor of about 30 to resolution. The action potentials of the Purkinje cells in the myocardium generate the time-quanta of the central nervous system as well as the mechanical scanning of the retinal image through the synchronic periodic variation of the tonus in the eye muscles. PMID- 16377060 TI - Reference values for the weight of freshly delivered term placentas and for placental weight-birth weight ratios. AB - BACKGROUND: There is evidence for a correlation between placental weight and future chronic disease, notably hypertension and diabetes. However, there are no reference scales for placentas that are readily weighed in the delivery room. METHODS: This cross-sectional study generated reference values for the weight of freshly delivered untrimmed placentas, and placental weight-birth weight (pw/bw) ratios from a database of 11,141 uncomplicated singleton term pregnancies (37-42 weeks). The data analysis followed stringent validated and state of the art methodological recommendations. A regression model was fitted to estimate the mean and standard deviation for placental weight and pw/bw ratios at each week of gestational age. RESULTS: Reference scales, percentile tables and regression equations are presented for placental weights according to the mode of delivery and for pw/bw ratios. Mean placental weight from vaginal deliveries was 76 g lighter than from Caesarean sections (545+/-107 g versus 621+/-139 g, respectively, P<0.05). Mean placental weight increased by 60 g from 37 to 42 weeks irrespective of the mode of delivery. The pw/bw ratio decreased from 17.6 to 15.6 between 37 and 42 weeks. CONCLUSION: For the first time, reference values for freshly delivered term placental weights depending on the mode of delivery were generated. In the light of growing evidence for a correlation of placental weight with chronic diseases in later life, these values provide the possibility to judge placentas at site for abnormalities in weight and to estimate the potential risks for chronic diseases in later life. PMID- 16377061 TI - Long-term outcome of vaginal sacrospinous colpopexy for marked uterovaginal and vault prolapse. AB - BACKGROUND: This study was carried out to evaluate the safety and long-term outcome of sacrospinous colpopexy in marked genital prolapse. SETTING: Gynaecology Department, Benenden Hospital, Kent, UK. METHODS: A prospective observational study was conducted between September 1993 and May 2000 on 305 women who underwent transvaginal sacrospinous colpopexy. The indications for surgery were marked vault prolapse in 43% and uterovaginal prolapse or enterocele in 57%. Patient follow up was at 6 weeks, 6 months, 1 year and then annually. Data was collected prospectively at the time of initial recruitment, during hospital stay and at the end of each follow up visit. RESULTS: Hysterectomy was performed in 117 patients and anterior colporrhaphy in 182. The mean operative time for the entire surgery was 65.6 min (S.D. 27.4, range 20-160 min) and estimated blood loss was 81.8 ml (S.D. 92, range 20-800 ml). After a mean follow up period of 57 months (range 24-84), vault support was maintained in 96%; recurrent vault prolapse occurred in 12 patients (4%) and the mean vaginal length at 1 and 5 years of follow up was 8+/-0.9 and 7.8+/-1.2 cm. Symptomatic cystocele occurred in 15 patients (5%). There were six recurrences of rectocele (2%) and there was no enterocele recurrence. Sexual function was maintained in all sexually active women and 43% reported improvement in sexual function. Out of 14 women who complained of fecal incontinence, 10 (71%) reported cure and 3 (21%) improved after surgery. CONCLUSIONS: Vaginal sacrospinous colpopexy is associated with a high long-term success rate in correcting upper genital prolapse. PMID- 16377062 TI - Recombinant gonadotrophins associated with GnRH antagonist (cetrorelix) in ovarian stimulation for ICSI: comparison of r-FSH alone and in combination with r LH. AB - OBJECTIVE: The objective was to verify the outcome of intracytoplasmic sperm injection (ICSI) with ovulation induction performed with GnRH antagonists, comparing the use of recombinant follicle-stimulating hormone (r-FSH) alone and in combination with recombinant luteinizing hormone (r-LH) in a prospective and randomized trial. STUDY DESIGN: Forty male-factor infertile normo-ovulatory patients undergoing ovarian stimulation for ICSI took part in the study. After initiating ovarian stimulation with only r-FSH, all patients were treated with GnRH antagonist (cetrorelix). When beginning cetrorelix administration, the patients were randomized into two groups: in group I, 20 patients continued to receive r-FSH alone and in group II, 20 patients received combined r-FSH and r LH. The number of metaphase II oocytes, estradiol concentration at the time of hCG administration, fertilization rate, grade 1 embryo rate, pregnancy rate per cycle, and implantation rate were measured. Results are expressed as mean+/-S.D. RESULTS: In group I, the women's age was 32.3+/-2.30 years, and FSH concentration was 7.8+/-1.7 IU/ml. In group II, the women's age was 32.2+/-2.46 years and FSH concentration was 7.5+/-1.7 IU/ml. The number of oocytes retrieved was 9.6+/-2.9 and the number of metaphase II oocytes was 6.7+/-2.2 in group I. In group II the number of retrieved oocytes were 9.9+/-2.6 and the number of metaphase II oocytes 6.9+/-2.1 (p>0.05). Estradiol concentration at the time of hCG was 4.6+/-1.8 nm/l in group I and 6.7+/-2.0 nm/l in group II (p<0.01). Fertilization rate was 73.0% in group I versus 78.2% in group II. In group I, we obtained 53.9% of grade 1 embryos versus 54.4% in group II (p>0.05). Pregnancy and implantation rates in group I were 30.0 and 16.7%, respectively and in group II 35.0 and 20.4%, respectively (p>0.05). CONCLUSIONS: The use of recombinant LH in addition to recombinant FSH may prevent a decrease in estradiol after GnRH antagonist administration, but does not influence positively the outcome of oocyte number, maturation, embryo quality, fertilization rate, pregnancy rate per cycle, and implantation rate. PMID- 16377063 TI - The impact of maternal cardioversion on fetal haemodynamics. PMID- 16377064 TI - Reproductive outcome after laparoscopic treatment of minimal and mild endometriosis using Helica Thermal Coagulator. AB - OBJECTIVE: To investigate the reproductive outcome following Helica Thermal Coagulator (Helica TC) in an infertile population with early-stage endometriosis. DESIGN: A clinical observational trial conducted in a NHS tertiary referral centre for treatment of endometriosis. Forty-four healthy infertile women (mean age 21 years, range 18-34 years) with laparoscopically confirmed diagnosis of minimal (stage I) or mild (stage II) endometriosis were included. All visible endometriotic lesions and adhesions were treated using Helica TC at the same time of diagnostic laparoscopy. Following laparoscopy, women attempted to conceive spontaneously and were followed for 12 months or until delivery. No hormonal therapy for endometriosis was given whatsoever. RESULTS: One woman was lost to follow-up and the remaining 43 women were considered in the final analysis. The cumulative pregnancy rate was 23.2% (10/43). All pregnancies occurred spontaneously and neither miscarriages nor ectopic pregnancies were observed. Pregnancy rate did not differ between women with stage I and those with stage II endometriosis. The time-to-pregnancy was similar in women with different stage endometriosis. No surgical complications were encountered. CONCLUSIONS: These findings suggest that Helica TC is a simple and safe conservative laparoscopic surgery for early-stage endometriosis. Helica TC appears to improve reproductive outcome in women with otherwise unexplained infertility. Large RCTs are needed to define these preliminary data. PMID- 16377065 TI - Safety and efficacy of low dose hCG for luteal support after triggering ovulation with a GnRH agonist in cases of polyfollicular development. AB - OBJECTIVE: The use of GnRH agonists instead of hCG to trigger ovulation seems to be an effective way to prevent subsequent hCG induced ovarian hyperstimulation in cases of polyfollicular development. But conflicting results are reported on the efficiency of subsequent luteal support using hCG and/or progesterone supplementation. STUDY DESIGN: We investigated the efficiency and safety of different luteal support regimes in low dose gonadotropin stimulation non-ivf cycles. A risk for an imminent ovarian hyperstimulation was assumed if preovulatory estradiol levels rose up higher than 700 pg/ml and more than 12 intermediate sized follicles (8-14 mm) were observed. Thirty-six women received 0.5mg Triptorelin subcutaneously to trigger the ovulation inducing LH surge. After randomization, luteal support regimes started on day 2 after the Triptorelin administration with injections every second day five times in all. Group (a) received 5 x 1000 IU hCG, group (b) received 5 x 500 IU hCG, and group (c) received 5 x 250 mg progesterone, intramuscularly. The monitoring of the ovulation period and the subsequent luteal phase included sonographic measurement of ovarian diameter and estimation of LH, FSH, estradiol and progesterone levels 10 and 34 h as well as 8 days after Triptorelin administration. RESULTS: We could prove ovulation in all women and did not find symptoms of ovarian hyperstimulation in any case. Midluteal controls showed extremely low gonadotropins in all groups indicating a long lasting pituitary down regulation after one injection of 0.5 mg Triptorelin. We found high normal sex steroid levels in both hCG groups. The progesterone group displayed a marked luteal phase defect with low levels of progesterone and estradiol in all cases. CONCLUSION: The use of GnRH agonist in cases of polyfollicular development is capable to induce ovulation without a subsequent ovarian enlargement and/or any sign of hyperstimulation syndrome. Luteal support by low dose hCG does not counteract the benefit of GnRH agonist in preventing an ovarian hyperstimulation syndrome, but seems to remedy at least in part the possible deleterious effects of GnRH agonists on luteal functionality. PMID- 16377066 TI - Risk factors of preterm delivery at less than 35 weeks in patients with renal transplant. AB - BACKGROUND: To identify prenatal events associated with preterm delivery at less than 35 weeks of gestation in women with renal transplant. METHODS: A case control study of 53 pregnancies in 42 renal transplant recipients, at a single center from 1984 to 2003 was analyzed. Preterm delivery cases (n=23) at less than 35 weeks of gestation were compared with the controls (n=30). RESULTS: Preterm delivery at less than 35 weeks of gestation occurred in 23 cases (43.4%). Hypertension (> or =140/90 mmHg) prior to pregnancy (odds ratio (OR) 6.3, 95% confidence interval (CI) 1.0-38.6), proteinuria (> or =0.3g/day) prior to delivery (OR 11.7, CI 2.7-51.8) and serum creatinine (> or =1.5mg/dl) prior to delivery (OR 4.4, CI 1.0-19.5) were significantly associated with increased risk of preterm delivery. Perinatal or neonatal deaths were not found. Fetal anomaly was seen in one case (polydactyly), and periventricular leukomalacia was found in two cases. CONCLUSION: In this case-control study, hypertension prior to pregnancy, proteinuria and serum creatinine (> or =1.5mg/dl) prior to delivery were related to the occurrence of preterm delivery at less than 35 weeks in renal transplant pregnancies. PMID- 16377067 TI - Vaginal douching and associated factors among married women attending a family planning clinic or a gynecology clinic. AB - OBJECTIVE: The aim of this study was to determine the vaginal douching habits and associated factors of women attending two different healthcare clinics. STUDY DESIGN: This cross-sectional study was conducted between 1 and 31 May 2004. All participants were women (n=635) who attended either a university hospital gynecology clinic or a primary health care center family planning unit. One-way ANOVA, chi-squared test, and binary and multiple logistic regression analyses were used for the statistical evaluation of data. RESULTS: The mean age of the participants was 36.90+/-10.72 years (range: 18-75). Half of the participants believed vaginal douching had a positive effect on health. They believed vaginal douching demonstrates cleanliness, prevents infections and pregnancy, removes sperm following intercourse, a necessity of Islamic doctrine, and reduces symptoms like discharge, unpleasant odor, etc. Of the women, 50.2% performed vaginal douching. Vaginal douching was associated with age, education level, type of dwelling, working outside of the home, age at marriage, age at birth of first child, parity, spontaneous abortion, history of pelvic inflammatory disease, use of contraceptives, and attending a healthcare clinic. CONCLUSION: Douching is a common habit among Turkish women. Many women are not aware of the harmful effects of douching. Public health and health professionals should monitor more closely this traditional habit in Turkey. PMID- 16377068 TI - A very low transverse uterine incision. PMID- 16377069 TI - Preferential hydration and solubility of proteins in aqueous solutions of polyethylene glycol. AB - This paper is focused on the local composition around a protein molecule in aqueous mixtures containing polyethylene glycol (PEG) and the solubility of proteins in water+PEG mixed solvents. Experimental data from literature regarding the preferential binding parameter were used to calculate the excesses (or deficits) of water and PEG in the vicinity of beta-lactoglobulin, bovine serum albumin, lysozyme, chymotrypsinogen and ribonuclease A. It was concluded that the protein molecule is preferentially hydrated in all cases (for all proteins and PEGs investigated). The excesses of water and deficits of PEG in the vicinity of a protein molecule could be explained by a steric exclusion mechanism, i.e. the large difference in the sizes of water and PEG molecules. The solubility of different proteins in water+PEG mixed solvent was expressed in terms of the preferential binding parameter. The slope of the logarithm of protein (lysozyme, beta-lactoglobulin and bovine serum albumin) solubility versus the PEG concentration could be predicted on the basis of experimental data regarding the preferential binding parameter. For all the cases considered (various proteins, various PEGs molecular weights and various pHs), our theory predicted that PEG acts as a salting-out agent, conclusion in full agreement with experimental observations. The predicted slopes were compared with experimental values and while in some cases good agreement was found, in other cases the agreement was less satisfactory. Because the established equation is a rigorous thermodynamic one, the disagreement might occur because the experimental results used for the solubility and/or the preferential binding parameter do not correspond to thermodynamic equilibrium. PMID- 16377070 TI - Optimization of Fenton's oxidation of chemical laboratory wastewaters using the response surface methodology. AB - Establishing a treatment process for practical and economic disposal of laboratory wastewaters has become an urgent environmental concern of the Department of Chemical Engineering of the Universidade Estadual de Maringa (State University of Maringa), Brazil. Fenton and related reactions are potentially useful oxidation processes for destroying toxic organic compounds in water. In these reactions, hydrogen peroxide is combined with ferrous or ferric iron in the presence or absence of light to generate hydroxyl radicals (.OH). The feasibility of Fenton's reagent to treat waste chemicals from an academic research laboratory was investigated in this study. A response surface methodology was applied to optimize the Fenton oxidation process conditions using chemical oxygen demand (COD) removal as the target parameter to optimize, and the reagent concentrations, as related to the initial concentration of organic matter in the effluent, and pH as the control factors to be optimized. Maximal COD removal (92.3%) was achieved when wastewater samples were treated at pH 4 in the presence of hydrogen peroxide and iron in the ratios [COD]:[H2O2]=1:9 and [H2O2]:[Fe2+]=4.5:1. Under these conditions, it was possible to obtain simultaneously maximal COD removal and minimal chemical sludge after treatment, which is a residue that needs further processing. PMID- 16377071 TI - Approximating pollution abatement costs via alternative specifications of a multi output production technology: a case of the US electric utility industry. AB - Using a panel of observations from the US electric utility industry, we analyze the output set frontiers that are produced by modeling the technology with different functional forms. Our specifications rely on a series of mapping regimes, based on the paths in which the outputs are scaled toward the frontier, and their outcomes are assessed against the general axiomatic framework of the production model. We demonstrate that different parameterization methodologies produce different empirical analogues of the output set and, as a consequence, can generate rather different estimates of the shadow prices of socially undesirable outputs, or 'bads,' as well. We benchmark our results by assessing the shadow price estimates, often interpreted as the opportunity cost of pollution reduction, against the market prices of pollution permits. The outcomes of our analysis suggest that the estimate of the output set boundary varies with the choice of a parameterization technique, which provides a possible explanation for the sizeable differences between the shadow prices of bads reported in the literature. We also demonstrate that among a variety of existing parametric methodologies no single technique is superior to all others. PMID- 16377072 TI - Characterization of haloacetic acid precursors in source water. AB - Raw water from the Bangkok (Thailand) main municipal water supply canal was examined for its natural organic composition by fractionation with adsorption resins. DAX-8 resin was the first resin employed to fractionate the hydrophobic fractions. Fractionation at neutral pH resulted in the separation of the hydrophobic neutral components; at a high pH level (approx. 10) separation of the hydrophobic base components occurred; and at a low pH level (approx. 2) the hydrophobic acid components were separated. AG-MP-50 cationic resin was then used to separate the hydrophilic base components, and WA-10, a weak anionic resin, was applied finally to fractionate the hydrophilic acid and neutral components. Subsequently, each fraction was tested for its chlorine disinfection by-product (DBP) formation potential. The HAA formation tests demonstrated that the various organic fractions had different reactivity levels for the formation of haloacetic acids (HAAs). For this source water, the hydrophilic neutral fraction dominated over the other five fractions in being the main organic component and the most significant precursor of HAAs formation. On the other hand, in terms of specific HAA formation potential (FP), the hydrophobic and hydrophilic base fractions were the most reactive precursors to the formation of HAAs. In all cases, the quantity of HAAs formed depended linearly upon the amount of organic constituents in the water sample. PMID- 16377074 TI - Biological significance of autoregulation through steady state analysis of genetic networks. AB - Autoregulation of regulatory proteins is a recurring theme in genetic networks. Autoregulation is an important component of a genetic regulatory network besides protein-protein and protein-DNA interactions, stoichiometry, multiple binding sites and cooperativity. Although the biological significance of autoregulation has been studied before, its significance in presence of other mechanisms is not clearly enumerated. We have analyzed at steady state the significance of autoregulation in presence of other molecular mechanisms by considering hypothetical genetic networks. We demonstrate that autoregulation of a regulatory protein can impart amplification to the response. Further, autoregulation of an activator binding to the DNA as a dimer can introduce bistability, thus forcing the system to reside in two distinct steady states. In combination with autoregulation, cooperative binding can further increase the sensitivity and can yield a highly ultrasensitive response. We conclude that autoregulation with the help of other molecular mechanisms can impart distinct system level properties such as amplification, sensitivity and bistability. The results are further discussed in relation to various examples of genetic networks that exist in biological systems. PMID- 16377073 TI - Retroperitoneoscopic versus open surgical radical nephrectomy for large renal cell carcinoma in clinical stage cT2 or cT3a: quality of life, pain and reconvalescence. AB - OBJECTIVES: To determine whether retroperitoneoscopic radical nephrectomy for large renal cell carcinoma in stage cT2 or cT3a is a feasible, safe and effective therapy option and if it shows any advantage regarding quality of life in comparison to open procedure. METHODS: 23 patients who underwent RPNx for tumor size greater than 7 cm (group 1) were matched and compared with 25 patients, who underwent ONx (group 2) for tumor with similar size characteristics. Patient and surgical data, QoL variables and complications were statistically analyzed. RESULTS: The median followup was 12 (range: 6-18) months for both groups. Group 1 had significantly (p < 0.001) less blood loss, shorter hospital stay, and shorter postoperative analgesic requirements. No conversion to open surgery was necessary, and no major complications requiring an invasive intervention occurred. Retroperitoneoscopic patients had significantly better QoL and pain scores postoperatively to 6 months (p < 0.001) and they return to baseline QoL status faster (p < 0.001). CONCLUSIONS: RPNx for large RCC in stage cT2 or cT3a is a safe and efficacious procedure with good short-term outcome results and significantly shorter recovery of QoL variables. PMID- 16377075 TI - Proceedings of the 3rd Dubrovnik International Conference on Multiple Sclerosis, Dubrovnik, Croatia, 19-21 May 2005. PMID- 16377076 TI - Cranberry phytochemical extracts induce cell cycle arrest and apoptosis in human MCF-7 breast cancer cells. AB - Breast cancer is the most commonly diagnosed cancer in women in the US and is one of the leading causes of death due to cancer. Epidemiological studies have consistently suggested the inverse association between cancer risk and intake of fruits and vegetables. These health benefits have been linked to the additive and synergistic combination of phytochemicals in fruits and vegetables. Cranberries have been shown to possess anti-carcinogenic activities such as inhibition of growth of several cancer cell lines, and inhibition of ornithine decarboxylase (ODC) activity in vitro. However, the molecular mechanisms of the anti-cancer properties of cranberry phytochemical extracts have not been completely understood. Our data showed that cranberry phytochemical extracts significantly inhibited human breast cancer MCF-7 cell proliferation at doses of 5 to 30mg/mL (P<0.05). Apoptotic induction in MCF-7 cells was observed in a dose-dependent manner after exposure to cranberry phytochemical extracts for 4h. Cranberry phytochemical extracts at a dose of 50mg/mL resulted in a 25% higher ratio of apoptotic cells to total cells as compared to the control groups (P<0.05). Cranberry phytochemical extracts at doses from 10 to 50mg/mL significantly arrested MCF-7 cells at G0/G1 phase (P<0.05). A constant increasing pattern of the G1/S index was observed in the cranberry extract treatment group while the G1/S ratio of the control group decreased concomitantly between 10 and 24h treatment. After 24-h exposure to cranberry extracts, the G1/S index of MCF-7 cells was approximately 6 times higher than that of the control group (P<0.05). These results suggest that cranberry phytochemical extracts possess the ability to suppress the proliferation of human breast cancer MCF-7 cells and this suppression is at least partly attributed to both the initiation of apoptosis and the G1 phase arrest. PMID- 16377077 TI - Single nucleotide polymorphisms in ABCC2 and ABCB1 genes and their clinical impact in physiology and drug response. AB - Among the ABC proteins, some members including ABCB1, ABCC1, ABCC2 and ABCG2 are believed to contribute to multidrug resistance of cancer chemotherapy. In addition, the broad substrate-specificity and apical localization of the ABCB1 and ABCC2 in mucosal epithelium of intestine and hepatocyte give them a protective role against xenobiotics. The inter-individual variations in activity and expression levels of ABCB1 and ABCC2, thus, might affect on drug response and response to toxic substrates. In this review, I focus on (1) physiological and toxicological relevance of ABCB1 and ABCC2, and on (2) genetic variations of ABCB1 and ABCC2 genes and their association with biochemical function, expression level and tumor incidence. PMID- 16377078 TI - Establishment of a highly tumorigenic LNCaP cell line having inflammatory cytokine resistance. AB - Human androgen-dependent prostate cancer LNCaP cells are low tumorigenic even in immunodeficient mice and were killed by the synergistic effect of inflammatory cytokines, IL-beta and IL-6. To establish a highly tumorigenic LNCaP cell line, we isolated the cytokine-resistant LNCaP-CR cell line and examined the phenotypes. The parental LNCaP cells were induced to commit apoptosis by the addition of IL-1beta and IL-6, but LNCaP-CR cells showed strong resistance against the cytokine action. However, LNCaP-CR cells did not exhibit any resistance to various antitumor drugs investigated. While LNCaP cells formed only palpable tumors in SCID mice, LNCaP-CR cells readily made tumors and their growth was significantly higher than that of LNCaP cells. Moreover, LNCaP tumor-bearing mice gained the weight gradually, but LNCaP-CR tumor-bearing mice significantly lost their body weight. LNCaP-CR cells still responded to androgen action and expressed AR, erbB2, IL-1R, IL-6R, gp130, STAT3, p21, Bcl-2 and caspase-3 as well as LNCaP cells. These results indicate that LNCaP-CR cell line is a new type of tumorigenic LNCaP cell lines and should be useful for identifying responsible genes of tumorigenicity, cytokine resistance, and also cachexia. PMID- 16377079 TI - An echo-contrast agent, Levovist, lowers the ultrasound intensity required to induce apoptosis of human leukemia cells. AB - To verify the effect of echo-contrast agent (ECA) on apoptosis induced by ultrasound, leukemia cell lines (Jurkat, Molt-4 and U937) were sonicated at intensities previously shown to induce optimal apoptosis with or without Levovist, an ECA. The results showed that loss of viability and apoptosis can be induced in all three cell lines, apoptosis highest with Molt-4, based on viability and DNA fragmentation assay. Such finding was supported by corresponding increase of cells with low mitochondrial membrane potential, high superoxide production, increased intracellular calcium concentration, and phosphorylation of histone H2AX after sonication. Optimal ultrasound condition was 0.3W/cm(2), 1MHz, 10% duty factor pulsed at 100Hz; but in the presence of Levovist, an apparent shift of cell killing induction was observed at 0.2W/cm(2). While these results further confirmed previous findings on ultrasound-induced apoptosis, they also suggest that use of an enhancing factor, such as addition of ECA, may be useful in cancer therapy when a much lower intensity is desired. PMID- 16377080 TI - The anti-obesity agent Orlistat is associated to increase in colonic preneoplastic markers in rats treated with a chemical carcinogen. AB - Orlistat is an anti-obesity agent that increases the fecal fat excretion, which promotes colon carcinogenesis. Therefore, the present study was designed to verify the effects of Orlistat on the formation of rat colonic aberrant crypt foci (ACF) and cell proliferation evaluated by the PCNA method. Male Wistar rats received either a standard diet or a high fat diet (HFD), supplemented or not with Orlistat (200mg/kg chow) and two doses of the carcinogen dimethyl-hydrazine (25mg/Kg). After 30 days, Orlistat was associated to a significant increase in the number of colonic ACFs and cell proliferation in DMH-treated animals, independently of the HFD. PMID- 16377081 TI - L1 on ovarian carcinoma cells is a binding partner for Neuropilin-1 on mesothelial cells. AB - The progression of ovarian cancer is driven by a variety of cellular factors that are incompletely understood. Binding of tumor cells to normal cells and to soluble factors influence tumor growth, angiogenesis and the stimulation of vascular permeability leading to ascites production. L1 adhesion molecule is overexpressed in ovarian carcinoma and is associated with bad prognosis. One receptor for L1 is Neuropilin-1 (NRP-1) that is also known as a receptor for VEGF(165). In the nervous system a complex of NRP-1 and L1 transmits signals by the neurorepellant Sem3A that is critical for the control of neurite outgrowth. NRP-1 has also been detected in human carcinomas but its function remains unknown. Here, we have examined NRP-1 expression in ovarian carcinoma cell lines and tissue. We report that little NRP-1 protein was detected in primary ovarian carcinoma tissues or established cell lines although mRNA for soluble and transmembrane NRP-1 were detected by RT-PCR. Instead, we observed strong expression of NRP-1 in mesothelial cells, which form the lining of the peritoneum. NRP-1 could serve as an isolation marker for primary mesothelial cells present in ascites fluid. We demonstrate that ovarian cancer cells expressing L1 can bind to NRP-1 overexpressing cells and mesothelial cells. Likewise, soluble L1 isolated from ascites of patients or produced as a fusion protein could bind to NRP-1 overexpressing cells and a direct interaction was demonstrated at the protein level. These findings suggest that L1 can support the binding of ovarian carcinoma cells to mesothelial cells via NRP-1. The L1-NRP-1 binding pathway could contribute to the growth of ovarian carcinomas and to reciprocal signalling between mesothelial cells and tumors. PMID- 16377082 TI - Cytochrome P450 2A6 polymorphism in nasopharyngeal carcinoma. AB - Nitrosamine has been identified as a carcinogen for nasopharyngeal carcinoma (NPC). Here, we investigated if a nitrosamine metabolizing gene, cytochrome P450 2A6 (CYP2A6) played an important role in NPC development. Relationships between the disease and the CYP2A6 were studied in 74 NPC patients and 137 age-matched healthy controls by using PCR-RFLP assay to distinguish between a wide type allele, *1A, and two mutant alleles, *1B and *4C. Overall, a significant association between CYP2A6 polymorphism and NPC development was observed (P<0.05). Individual with mutant alleles had an increased risk for NPC when compared to those with *1A/*1A (OR=2.37, 95% CI=1.27-4.46). In addition, males who carried mutant alleles of CYP2A6 had a fivefold increased risk for NPC when compared with those who carried *1A/*1A genotype (OR=5.02, 95% CI=1.82-14.14). It is thus suggested that CYP2A6 polymorphism may play a crucial role in NPC susceptibility and it may be used as a risk marker for NPC. PMID- 16377083 TI - Differential effects of Stat3 inhibition in sparse vs confluent normal and breast cancer cells. AB - The signal transducer and activator of transcription-3 (Stat3) is persistently activated in many cancers such as cancer of the breast and is required for transformation by a number of oncogenes. Signaling through Stat3 is determined by a key phosphorylation at tyr-705. We previously demonstrated that cell-to-cell adhesion brought about through cell aggregation or confluence of cultured cells causes a dramatic increase in Stat3 tyr705 phosphorylation and consequently Stat3 activity in both normal and tumor cells. To examine the role of Stat3 at specific time-points relative to confluence, we used two different approaches of Stat3 inhibition: (1). Introduction of high levels of peptide analogues, which block the Stat3-SH2 domain, to inhibit Stat3 binding to and phosphorylation by growth factor receptors. (2). Treatment with two platinum compounds which bind the Stat3 protein and inhibit its activity without affecting its phosphorylation directly. The results demonstrate that Stat3 downregulation in vSrc transformed NIH3T3 cells or in breast cancer lines harboring activated Src induces apoptosis, which is evident at all densities but is more pronounced at post-confluence. In normal cells on the other hand, Stat3 inhibition at post-confluence caused apoptosis while in sparsely growing cells it induced merely a growth retardation. PMID- 16377084 TI - Graft derived cells with double nuclei in the penumbral region of experimental brain trauma. AB - Recent in vitro studies showed that stem cells might fuse with mature cells or each other; however, there is no in vivo evidence for this phenomenon in the cerebral cortex. Our goal was to find evidence for cell fusion in a model of traumatic brain injury followed by grafting of embryonic cortical cells. Cold lesion protocol was applied to induce lesion of the motor cortex in adult male rats. Six days later we grafted a suspension of freshly isolated rat brain cortical cells of early embryonic stage (E14) into the penumbra area of the lesion. The grafted cell nuclei were labelled with bromodeoxyuridine (BrDU). Six days after transplantation 4,328 BrDU positive cells were observed in nine animals. 89.5% of these cells had cytoplasmic staining probably representing dead or phagocyted grafted cells. Ten percent of surviving BrDU positive cells had only one BrDU positive nucleus and negative cytoplasm, while 0.5% had two distinct nuclei, one was unlabelled and one was BrDU positive. These cells were similar in appearance and size to the astrocytes in the vicinity and expressed the astocyte specific glial fibrillaly acidic protein. Thus, these cells showed a possible sign of cell fusion in the penumbral region of the injured brain. PMID- 16377085 TI - Increases in TH immunoreactivity, neuromelanin and degeneration in the substantia nigra of middle aged mice. AB - The dopaminergic (DArgic) neurons in the substantia nigra (SN) are particularly vulnerable to oxidative stress and during aging. The present study was undertaken in order to determine whether aging is associated with changes in the DA synthesizing enzyme tyrosine hydroxylase (TH) as early as middle age by comparing 7- and 50-week-old mice. Quantitative analysis, performed by measuring the density of TH-immunopositive neurons, revealed that in the older animals, the number of DArgic neurons was decreased by 10% while TH immunodensity was 24 +/- 3% higher compared to the younger animals. Based on Masson-Fontana staining for neuromelanin (NM), the number of NM-containing neurons in the SN and the volume of NM per NM-positive neurons in the older animals were 5- and 11.6 +/- 0.1-fold higher, respectively. The silver stain-positive fibers, indicative of degeneration, were higher in the SN and striatum of the older animals, with the optical density 3.3 +/- 0.1- and 5.4 +/- 0.2-fold of the younger animals. The present study demonstrates that aging is associated with changes in the DA synthesizing enzyme TH as early as middle age and that this is associated with dramatic increases in the number of NM-containing neurons, volume of NM per cell, and degeneration. PMID- 16377086 TI - Role of Staufen in dendritic mRNA transport and its modulation. AB - Staufen is a double-stranded RNA-binding protein and a core component in various RNP complexes or RNA granules, and plays an important role in dendritic mRNA transport. In this study, a ribosomal marker and a dominant-negative form of Staufen (stau-RBD), containing the RNA-binding domains, but lacking a microtubule association domain, was used to determine the role of Staufen in dendritic mRNA transport. The results showed that the overexpression of stau-RBD significantly decreased the levels of ribosomal staining in the dendrites, which was illustrated by Y10B immunostaining. In contrast, the overexpression of Staufen increased the ribosomal level. The regulatory mechanisms of the dendritic mRNA transport were examined using a GFP-tagged Staufen (GFP-Stau), which was produced by means of a Sindbis viral expression system. Depolarization increased the amount of Staufen-containing the RNP complexes and endogenous Staufen in the dendrites. This increase was independent of protein synthesis. This suggests that dendritic mRNA transport is mediated via Staufen, and is regulated by the neuronal activity. PMID- 16377087 TI - Effects of neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one on ethanol-mediated paired-pulse depression of population spikes in the CA1 region of rat hippocampal slices. AB - While it is known that ethanol augments GABA-A receptor mediated inhibition in the central nervous system (CNS), demonstrating direct effects of ethanol on GABA transmission has been difficult in brain slices, suggesting that these preparations may lack factors that are required for ethanol's actions. Recent studies indicate that the GABA-enhancing neurosteroid 3alpha-hydroxy-5alpha pregnan-20-one (3alpha5alphaP) mediates at least some effects of ethanol in the CNS. In the CA1 region of rat hippocampal slices, we found that 60mM ethanol failed to alter paired pulse depression (PDD) of population spikes (PSs) when paired stimuli were delivered to the Schaffer collateral pathway at an interval of 21ms. Following 2-h preincubation of slices with 100nM 3alpha5alphaP, however, ethanol augmented PS PPD. This effect was not observed in the presence of picrotoxin, a GABA-A receptor antagonist, or ADVASEP-7, a beta-cyclodextrin that binds 3alpha5alphaP. These results indicate that 3alpha5alphaP modulates the inhibitory effects of ethanol on hippocampal excitability via GABA-A receptors. PMID- 16377088 TI - Opioid-induced regulation of gene expression in PC12 cells stably transfected with mu-opioid receptor. AB - It has been postulated that opiates induce addictive behaviour via changes in gene expression. PC12 cells were stably transfected with the recombinant human mu opioid receptor (MOR) to study opioid-induced gene expression. Expression was verified by binding assay, immunocytochemistry, and immunblotting experiments. Forskolin-induced cAMP formation was inhibited by [D-Ala(2), N-Me-Phe(4), Gly(5) ol]-enkephalin (DAMGO 1 microM), a specific MOR agonist. This effect was completely antagonized by naloxone. By using cDNA arrays, including approximately 1,200 well-defined genes normally expressed in neural tissue, we monitored semi quantitative changes in gene expression after 3 h short-term exposure to DAMGO. Incubation with DAMGO increased mRNA levels for 13 genes and down-regulated 13 other genes. Annexin V, RGS4 and CREB genes showed pronounced increase in expression after stimulation with DAMGO. Quantitative RT-PCR confirmed that DAMGO increased mRNA levels of Annexin V, an apoptosis-induced gene. We suggest that the PC12 cell transfected with the recombinant human MOR is a useful tool for identification of opioid-induced genes that may provide information on opiate effects of relevance for dependence. PMID- 16377089 TI - Cross-modal innervation of primary visual cortex by auditory fibers in congenitally anophthalmic mice. AB - Auditory-visual cross-modal innervation was examined in control (sighted, ZRDCT N) and congenitally anophthalmic (eyeless, ZRDCT-AN) mice using electrophysiological recording and pathway tracing with carbocyanine dyes. Electrophysiological data demonstrate that the primary visual cortex of congenitally eyeless, blind, mice receives auditory stimuli. Neuroanatomical data demonstrate a direct connection between the inferior colliculus (IC) and visual cortex. Our experiments provide new information about how the brain adapts to the loss of sight. PMID- 16377090 TI - World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guidelines for evaluating the efficacy of acaricides against ticks (Ixodidae) on ruminants. AB - These guidelines have been prepared to assist in the planning, conduct and interpretation of studies for the assessment of the efficacy of acaricides (excluding vaccines and other bio-control agents) against single and multi-host ticks (Ixodidae) on ruminants. Information is provided on the selection of animals, dose determination, dose confirmation and field studies, record keeping and result interpretation. The use of pen facilities is advocated for dose determination and confirmation studies for defining therapeutic and persistent efficacy. A minimum of two studies per tick species for which claims are sought is recommended for each dose determination and dose confirmation investigation. If dose confirmation studies demonstrate greater than 95% efficacy the sponsor may proceed to field studies, where a minimum of two studies per geographical location is preferred to confirm the therapeutic and persistent efficacy under field conditions. If dose confirmation studies demonstrate less than 95% efficacy then longer-term field studies can be conducted over two tick seasons with a minimum of two studies per geographical location. These studies can incorporate other control methods such as tick vaccines, to demonstrate stable long-term tick management. Specific advice is also given on conducting studies with paralysis ticks. These guidelines are also intended to assist investigators on how to conduct specific experiments, to provide specific information for registration authorities involved in the decision-making process, to assist in the approval and registration of new acaricides, and to facilitate the worldwide adoption of standard procedures. PMID- 16377091 TI - Lipid rafts are involved in C95 (4,8) agrin fragment-induced acetylcholine receptor clustering. AB - During development of the neuromuscular junction, high densities of acetylcholine receptors accumulate beneath the overlying nerve terminal. A defining feature of mature synapses is the sharp demarcation of acetylcholine receptor density, which is approximately 1000-fold higher in the postsynaptic as compared with the contiguous extrasynaptic muscle membrane. These high densities of receptors accumulate by at least four mechanisms, re-distribution of existing surface receptors, local synthesis of new receptors, decreased turnover of synaptic receptors, and limitation of diffusion of sub-neural, aggregated receptors. The limitation of receptor diffusion within the membrane is likely in part due to the anchoring of acetylcholine receptor complexes to components of the cytoskeleton. Here we have tested the idea that lipid rafts--mobile, cholesterol enriched microdomains within the lipid bilayer--are another mechanism by which acetylcholine receptors are clustered in the postsynaptic apparatus. Using mouse C2C12 cells, a muscle cell line, we show that a carboxy terminal 95 amino acid fragment [C95 (4,8)] of the extracellular matrix molecule agrin that is essential for nerve-induced postsynaptic differentiation, promotes the redistribution of acetylcholine receptors into lipid rafts. Disruption of lipid rafts before agrin treatment largely inhibits de novo agrin-induced acetylcholine receptor clustering. Moreover, mature acetylcholine receptor clusters are destabilized if lipid rafts are disrupted. These results show that lipid rafts are important in both the initial clustering and later stabilization of agrin-induced acetylcholine receptor clusters and also suggest that lipid rafts may contribute to the postsynaptic localization of acetylcholine receptors in vivo. PMID- 16377092 TI - Dynamic changes in orbitofrontal neuronal activity in rats during opiate administration and withdrawal. AB - The orbitofrontal cortex is involved in the reinforcing effects of drugs of abuse. However, how the dynamic activity in OFC changes during opiate administration and withdrawal period has not been investigated. We first tested the effects of opiates and drug craving with the conditioned place preference paradigm, using manganese-enhanced magnetic resonance imaging and traditional electroencephalograph recording techniques in rats. T1-weighted 2D MRI (4.7 T) was used after unilateral injection of MnCl(2) (200 nL, 80 mM) into the right orbitofrontal cortex. The manganese-enhanced magnetic resonance imaging data suggested that the OFC activity decreased during the opiate administration period but recovered increasingly during the withdrawal period. Also, we found decreases and increases in gamma-band (20-100 Hz) activity during the opiate administration and withdrawal period, respectively. Our results showed that orbitofrontal cortex activity decreased during morphine administration and then went up progressively over several days during withdrawal. The time course of the recovery of orbitofrontal activity from inhibition during the withdrawal period may be related to the experience of drug craving. PMID- 16377093 TI - Cocaine elicits action potential bursts in a central snail neuron: the role of delayed rectifying K+ current. AB - The effects of cocaine were studied in an identifiable RP4 neuron of the African snail, Achatina fulica Ferussac, using the two-electrode voltage-clamp method. The RP4 neuron generated spontaneous action potentials and bath application of cocaine (0.3-1 mM) reversibly elicited action potential bursts of the central RP4 neuron in a concentration-dependent manner. The action potential bursts were not blocked when neurons were immersed in high-Mg(2+)solution, Ca(2+)-free solution, nor after continuous perfusion with atropine, d-tubocurarine, propranolol, prazosin, haloperidol, or sulpiride. Similarly, the action potential bursts were not abolished by pretreatment with N-[2-(p-bromocinnamylamino)ethyl]-5 isoquinolinesulfonamide dihydrochloride, (9S,10S,12R)-2,3,9,10,11,12-hexahydro-10 hydroxy-9-methyl-1-oxo-9,12-epoxy-1H-diindolo[1,2,3-fg:3',2',1'-kl]pyrrolo[3,4 i][1,6]benzodiazocine-10-carboxylic acid hexyl ester or anisomycin. Injection of hyperpolarizing current at an intensity of greater than 2 nA effectively suppressed the cocaine-elicited action potential bursts and no postsynaptic potentials were observed under these conditions. These results suggest that the generation of action potential bursts elicited by cocaine was not due to (1) the synaptic effects of neurotransmitters, (2) the cholinergic, adrenergic or dopaminergic receptors of the excitable membrane, or (3) the cAMP second messengers and new protein synthesis of the RP4 neuron. Notably, the induction of action potential bursts was blocked by pretreatment with 1-[6-[((17beta)-3 methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1H-pyrrole-2,5-dione. Voltage clamp studies conducted on the RP4 neuron revealed that cocaine at 0.3 mM decreased (1) the Ca(2+) current, (2) the delayed rectifying K(+) current, (3) the fast-inactivating K(+) current and (4) the Ca(2+)-activated K(+) current, but had no remarkable effects on the Na(+) current. Perfusion with Ca(2+)-free solution, which may abolish the Ca(2+) current and Ca(2+)-activated K(+) current, did not cause any bursts of action potentials in control RP4 neurons. Application of 4-aminopyridine, an inhibitor of fast-inactivating K(+) current, and paxilline, an inhibitor of Ca(2+)-activated K(+) current, failed to elicit action potential bursts, whereas tetraethylammonium chloride, a blocker of Ca(2+) activated K(+) current and delayed rectifying K(+) current, and tacrine, an inhibitor of delayed rectifying K(+) current, successfully elicited action potential bursts. Further, while 1-[6-[((17beta)-3-methoxyestra-1,3,5[10]-trien 17-yl)amino]hexyl]-1H-pyrrole-2,5-dione did not affect the delayed rectifying K(+) current of the RP4 neuron, 1-[6-[((17beta)-3-methoxyestra-1,3,5[10]-trien-17 yl)amino]hexyl]-1H-pyrrole-2,5-dione decreased the inhibitory effect of cocaine on the delayed rectifying K(+) current. It is concluded that cocaine elicits action potential bursts in the central snail RP4 neuron and that the effect is closely related to the inhibitory effects on the delayed rectifying K(+) current. PMID- 16377094 TI - Trait anxiety moderates the impact of performance pressure on salivary cortisol in everyday life. AB - Stress and negative affective states are associated with cortisol in everyday life. However, it remains unclear what types of stressors and which affective states yield these associations, and the effect of trait anxiety is unknown. This study investigates the associations of specific task-related stressors and negative affective states in everyday life with salivary cortisol, and explores the mediating and moderating role of state negative affect and trait anxiety, respectively. Salivary cortisol, subjective stress, and state negative affect were measured three times a day on 2 days in 71 participants in everyday life, using a handheld computer to collect self-reports and time stamps and an electronic device to monitor saliva sampling compliance. Stress measures comprised the experience of performance pressure and failure during daily tasks; measures of negative affect comprised worn-out, tense, unhappy, and angry. Effects were tested using multilevel fixed-occasion models. Momentary performance under pressure was related to higher momentary cortisol measures, while mean task failure was related to lower daily cortisol concentrations. The association of performance pressure with cortisol varied between subjects, and this variation was explained by trait anxiety, yielding stronger associations in participants scoring high on trait anxiety. No evidence was found for a mediating role of state negative affect. These results describe the well-documented associations of everyday stressors and affect with salivary cortisol more precisely, suggesting that performance pressure is a significant condition related to short-term changes in cortisol. Subjects scoring high on trait anxiety seem to process stress-relevant information in a way that amplifies the association of performance pressure with reactions of the hypothalamus-pituitary-adrenal axis. PMID- 16377095 TI - Association study of androgen receptor CAG repeat polymorphism and male violent criminal activity. AB - Androgens exert their effects primarily by stimulating androgen receptors (ARs) and androgen activity has been implicated in antisocial or violent criminal behaviour. Exon 1 of the AR gene contains a highly polymorphic glutamine (CAG) repeat sequence. We tested the hypothesis that shorter AR CAG repeat polymorphisms, which have a greater AR gene expression, are related to violent criminal activity in Chinese males using a sample of 146 extremely violent criminals and 108 normal controls. The results show no association between the AR repeat length and violent convicts, although we found that there are more violent criminal cases than control cases with the shorter (<17) AR gene trinucleotide repeat polymorphisms (7.5% vs. 1.9%; P=0.047). No correlation was found between the AR CAG repeat length and the first criminal record age in the violent criminal group. The mean AR repeat length is not significantly different between substance dependence cases and antisocial personality disorder diagnosis in the violent criminals and normal controls. Our findings suggest that the AR CAG repeat polymorphism does not play a major role in the susceptibility of male violent criminal activity. PMID- 16377096 TI - Mammalian oocyte polarity can be exploited for the automation of somatic cell nuclear transfer--in the development of a 'cloning biochip'. PMID- 16377097 TI - Sudden infant death syndrome (SIDS): microgravity and inadequate sensory stimulation. AB - In early gestation, the human foetus develops in a buoyant environment, which is similar to the near-weightlessness of space flight. After the 26th week of gestation, the foetus gradually becomes exposed to gravitational forces. The influence of fluid immersion declines as the weight of the foetus increases. In this way, the foetus adapts and trains for the gravity environment after birth. Failure of gravitational loading in the last trimester of pregnancy delays development and maintains the pathophysiological environment of microgravity as experienced by the astronaut in space flight. The deconditioning effects of microgravity during space flight are the reverse processes of intrauterine development after the 26th week when the foetus begins training body processes for adaptation to an earthly environment. Growth requires space and movement, which suggests that a growth-retarded foetus may have been deprived of the mechanical dimension of uterine wall pressure, and, in twins, the smaller sibling may have been deprived of space. The behaviour of a study group of sudden infant death syndrome infants suggested a continuation of the effects of the foetal akinesia syndrome during the third trimester period of gestation. NASA research into the pathophysiology of microgravity was based on a simple insight: that the physiological effects of human space travel were virtually identical to the adjustments the body makes when lying down. This is the same environment as that of the human foetus in the first 22 weeks of gestation after which the uterine environment becomes a prelude to adaptations to the force of gravity. PMID- 16377098 TI - Potential use of therapies for osteoporosis to treat neurological diseases of elderly people. PMID- 16377099 TI - Intrahepatic cholangiocarcinoma arising in cirrhotic liver frequently expressed p63-positive basal/stem-cell phenotype. AB - In general, intrahepatic cholangiocarcinoma (ICC) is not related to liver cirrhosis. However, a few cases have been reported in which ICC was accompanied by severe liver fibrosis. Some researchers have proposed that hepatocellular and cholangiocellular (HC-CC) carcinoma, an intermediate mixed phenotype possibly arising in cirrhotic liver, might originate from hepatic precursor cells. In the liver, hepatocytes and cholangiocytes form the epithelial element, but stromal and mesenchymal elements may be produced by hepatic stem cells. Based on these aspects, not only HC-CC, but also other combinations of cellular phenotypes, would cover all the cancers with stem cell features. In this study, which aimed at determining the characteristics of the ICC phenotype, we used immunohistochemistry to examine the expression of basal/stem-cell markers, i.e., p63 in ICC with and without liver cirrhosis, as well as the expressions of cytokeratin (CK) 34 beta E12, specific for the basal-cell marker, and c-kit, specific for the stem-cell marker. Aberrant p63 was frequently expressed in ICC arising in cirrhotic liver. This result suggests that ICC cancer cells originate from hepatic precursor cells with a hidden multi-differentiation potential. PMID- 16377100 TI - Isolated Rosai-Dorfman disease of intracranial meninges. AB - Rosai-Dorfman disease (RDD) is a non-neoplastic proliferative histiocytic disorder that primarily affects lymph nodes (sinus histiocytosis with massive lymphadenopathy). Primary RDD of the central nervous system is most uncommon. We report on a 35-year-old man with isolated RDD of the meninges overlying the left cerebral hemisphere. Presenting signs and symptoms included severe progressive ipsilateral headaches of 4 months duration, as well as laboratory evidence of mild non-specific systemic inflammatory reaction. On magnetic resonance imaging, the lesion was seen as a contrast-enhancing, plaque-like thickening of the dura mater over the left convexity,without impinging on adjacent bone or cerebral parenchyma. Meningeal biopsy revealed a mixed mononuclear infiltrate dominated by CD68(+), S100(+), CD1a(-) non-Langerhans type histiocytes on a background of fibrosis. Bacteria, in particular mycobacteria, and fungi were excluded with special stains. Extensive clinical workup, encompassing computed tomography of thoracal and abdominal organs, bone marrow biopsy, and bronchoalveolar lavage failed to reveal any extracranial involvement. Laboratory tests for autoimmunity, including C- and P-antineutrophil cytoplasmic antibodies, antinuclear antibody, and serum rheumatoid factor, were negative. Methylprednisolone therapy induced complete remission of symptoms, with the neuroradiologic status remaining unchanged on follow-up after 2 months. We discuss the complex clinicopathologic differential diagnosis and therapeutic issues of this rare condition. While the correct diagnosis of central nervous system RDD is unlikely to be established without invasive procedures (biopsy), a conservative therapeutic approach may be considered a legitimate option. PMID- 16377101 TI - Prize-based contingency management does not increase gambling. AB - A contingency management (CM) intervention that provides drug-abstinent patients a chance to win prizes of varying magnitudes is efficacious in retaining patients in treatment and reducing drug use. However, this intervention has been criticized as possibly increasing gambling because it contains an element of chance. Gambling behaviors before, during and 3 months after participation in a multi-site study of CM were compared for stimulant users randomly assigned to 12 weeks of standard care with (N=407) or without (N=396) prize-based CM. Among study participants enrolled in outpatient non-methadone drug abuse treatment (N=415), 26% reported gambling during the observation period, and this rate was 37% among participants (N=388) enrolled in methadone maintenance programs. No differences in gambling over time were noted between those assigned to the prize CM versus standard care conditions, indicating that this prize CM procedure does not adversely impact gambling behavior among stimulant abusers. PMID- 16377102 TI - Epidermal growth factor receptor (EGFR) signaling in cancer. AB - The epidermal growth factor receptor (EGFR) belongs to the ErbB family of receptor tyrosine kinases (RTK). These trans-membrane proteins are activated following binding with peptide growth factors of the EGF-family of proteins. Evidence suggests that the EGFR is involved in the pathogenesis and progression of different carcinoma types. The EGFR and EGF-like peptides are often over expressed in human carcinomas, and in vivo and in vitro studies have shown that these proteins are able to induce cell transformation. Amplification of the EGFR gene and mutations of the EGFR tyrosine kinase domain have been recently demonstrated to occur in carcinoma patients. Interestingly, both these genetic alterations of the EGFR are correlated with high probability to respond to anti EGFR agents. However, ErbB proteins and their ligands form a complex system in which the interactions occurring between receptors and ligands affect the type and the duration of the intracellular signals that derive from receptor activation. In fact, proteins of the ErbB family form either homo- or hetero dimers following ligand binding, each dimer showing different affinity for ligands and different signaling properties. In this regard, evidence suggests that cooperation of multiple ErbB receptors and cognate ligands is necessary to induce cell transformation. In particular, the growth and the survival of carcinoma cells appear to be sustained by a network of receptors/ligands of the ErbB family. This phenomenon is also important for therapeutic approaches, since the response to anti-EGFR agents might depend on the total level of expression of ErbB receptors and ligands in tumor cells. PMID- 16377103 TI - In vitro degradation of equine keratin by dermatophytes and other keratinophilic fungi. AB - Keratinolytic properties of two dermatophytes (Microsporum gypseum, Trichophyton mentagrophytes) and three moulds (Scopulariopsis brevicaulis, Alternaria alternata, Geotrichum candidum) isolated from diseased equine hooves were examined to improve the understanding of pathogenic mechanisms leading to equine onychomycosis. Equine hoof horn material and skin, as well as hoof keratin and dermal keratin extracted from corresponding tissues, were used as sole carbon and nitrogen sources in five test tubes for each fungus. Within 18 days, supernatants of all tubes were repeatedly examined for keratinolytic activity by SDS-PAGE and Western blot analysis. In addition, fungal growth rates were determined to identify the preferred tissue of the individual fungi. Among the fungi examined, M. gypseum was the most keratinolytic species, followed by T. mentagrophytes and S. brevicaulis. In the concentration applied, the moulds A. alternata and G. candidum showed minimal keratinolytic activity. With respect to growth rates, M. gypseum favoured hoof horn material, S. brevicaulis and G. candidum preferred skin as a keratin source, whereas for the other two fungi no clear preference was detectable. PMID- 16377104 TI - Obstructive uropathy. AB - Routine second trimester ultrasound screening has resulted in more infants diagnosed with antenatal hydronephrosis. Current recommendations suggest postnatal evaluation of all infants with a renal pelvic diameter >5 mm with ultrasound and voiding cystourethrogram (VCUG.) There are many etiologies of obstructive uropathy including ureteropelvic junction (UPJ) obstruction, ureterovesical junction (UVJ) obstruction, posterior urethral valves (PUV), prune belly syndrome, and vesicoureteral reflux (VUR). Obstructive uropathy can result in tubular damage and decreased nephron number. Tubular damage can result in sodium wasting, hyperkalemic acidosis, and nephrogenic diabetes insipidus. Most patients do not require renal replacement therapy in the neonatal period; however, chronic renal insufficiency can occur if the neonate has a significant reduction in nephron number or progressive renal damage from obstruction or infection. PMID- 16377105 TI - Effect of milling and sieving on functionality of dry powder inhalation products. AB - Alpha-lactose monohydrate is the standard excipient used as diluent or carrier in dry powder inhaler (DPI) formulations. Earlier studies have already revealed that raw materials for the production of inhalation grade lactose have to be carefully selected in order to avoid batch-to-batch variability. In the present study, the effect of milling and milling intensity on the flow properties and the physico chemical characteristics of lactose crystals has been determined. The milled lactoses were then further processed by sieving to give lactose qualities with identical size distribution data, but different batch history (non-milled and milled at different conditions). These were then used to manufacture low concentration (0.25%) drug blends with the model drugs salbutamol sulphate (SBS) and beclometasonedipropionate (BDP); the blends were analysed with a Multistage Liquid Impinger (MLI) after delivery from an Easyhaler and an Aerolizer device. It could be shown that gentle milling already results in surface defects on the lactose crystal which are further enhanced by using a higher milling intensity. Produced fine lactose particles during the milling process strongly adhere to the lactose surface and cannot be removed by compressed air which is used for the particle sizing. By trend, a higher milling intensity resulted in higher fine particle fractions (FPF) with both devices. Also, SBS was found to generally give higher fine particle fractions than BDP, independent from the device used. In conclusion, lactose pre-treatment by gentle or strong milling affects the carrier surface and thereby the aerosolization properties of drug/lactose blends produced. PMID- 16377106 TI - Self-assembled drug delivery systems. 1. Properties and in vitro/in vivo behavior of acyclovir self-assembled nanoparticles (SAN). AB - Self-assembled drug delivery systems (SADDS) were designed in the paper. They can be prepared from the amphiphilic conjugates of hydrophilic drugs and lipids through self-assembling into small-scale aggregates in aqueous media. The outstanding characteristic of SADDS is that they are nearly wholly composed of amphiphilic prodrugs. The self-assembled nanoparticles (SAN) as one of SADDS had been prepared from the lipid derivative of acyclovir (SGSA) in the previous paper. They were further studied on the properties and the in vitro/in vivo behavior in this paper. The SAN kept the physical state stable upon centrifugation or some additives including some inorganic salts, alkaline solutions, surfactants and liposomes except for HCl solution, CaCl(2) solution and animal plasma. Autoclave and bath heat for sterilization hardly influenced the SAN. However, gamma-irradiation strongly destroyed the structure of SAN and SGSA was degraded. SGSA in SAN showed good stability in weak acidic or neutral buffers although it was very sensitive to alkaline solutions and carboxylester enzymes, the half-lives (t(1/2)) of which in the buffer at pH 7.4, the alkaline solution at pH 12.0, pig liver carboxylester enzyme solution, rabbit plasma, and rabbit liver tissue homogenate were 495, 21, 4.7, 25 and 8.7 h, respectively. Compared with SGSA in a disordered state, the specific bilayer structures of SAN could protect SGSA from hydrolysis through hiding the sensitive ester bonds. The SAN showed hemolytic action because the amphiphilic SGSA could insert into rabbit erythrocyte membranes. Both the high concentration of SGSA in samples and the long incubation time improved hemolysis. No hemolysis was observed if the additional volume of the SAN was less than 10% of rabbit whole blood in spite of the high concentration of SGSA. Plasma proteins could interfere the interaction between the SAN and erythrocytes by binding the SAN. The in vitro antiviral activity of acyclovir SAN was limited possibly because of the weak hydrolysis of SGSA in Vero cells, and the SAN showed a little cell toxicity possible due to the amphiphilicity of SGSA. A macrophage cell line of QXMSC1 cells showed uptake of the SAN but not significantly. The SAN were rapidly removed from blood circulation after bolus iv administration to rabbits with the very short distribution t(1/2) (1.5 min) and the elimination t(1/2) (47 min). The SAN were mainly distributed in liver, spleen and lung after iv administration, and SGSA was eliminated slowly in these tissues (t(1/2), about 7 h). It would appear that the nanosized SAN were trapped by the mononuclear phagocyte system. SADDS including SAN combine prodrugs, molecular self-assembly with nanotechnology, and hopefully become novel drug delivery approaches. PMID- 16377107 TI - Formulation and biological evaluation of glimepiride-cyclodextrin-polymer systems. AB - Glimepiride is one of the third generation sulfonylureas used for treatment of type 2 diabetes. Poor aqueous solubility and slow dissolution rate of the drug lead to irreproducible clinical response or therapeutic failure in some cases due to subtherapeutic plasma drug levels. Consequently, the rationale of this study was to improve the biological performance of this drug through enhancing its solubility and dissolution rate. Inclusion complexes of glimepiride in beta cyclodextrin (beta-CyD), hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) and sulfobutylether-beta-cyclodextrin (SBE-beta-CyD), with or without water soluble polymers were prepared by the kneading method. Binary systems were characterized by thermogravimetric analysis, IR spectroscopy and X-ray diffractometry. Phase solubility diagrams revealed increase in solubility of the drug upon cyclodextrin addition, showing A(p) type plot indicating high order complexation. All the ternary systems containing beta-CyD or HP-beta-CyD showed higher dissolution efficiency compared to the corresponding binary systems. The hypoglycemic effect of the most rapidly dissolving ternary system of glimepiride-HP-beta-CyD-PEG 4000 was evaluated after oral administration in diabetic rats by measuring blood glucose levels. The results indicated that this ternary system improves significantly the therapeutic efficacy of the drug. In conclusion, the association of water soluble polymers with glimepiride-CyD systems leads to great enhancement in dissolution rate, increased duration of action and improvement of therapeutic efficacy of the drug. PMID- 16377108 TI - Influence of absorption enhancers on the pharmacokinetic properties of non-oral beta-lactam-cefpirom using the rabbit (Chinchilla) in vivo model. AB - The oral application is the application of the first choice for drug administration. A lot of drugs exhibit relatively low bioavailability. This may be caused by binding of the drug in the gastro-intestinal tract, by poor penetration of the intestinal mucose or by highly hydrophilic properties. Therefore, problem drugs were only used for i.v. administration (intravenously) or for i.m. administration (intramuscularly). In the present study, cefpirom was investigated as a model substance. Cefpirom (Cp) is a semi-synthetic amino-2 thiazolyl-methoxyimino cephalosporin. It exhibits highly hydrophilic properties (P(ow)=0.02+/-0.01) and a very low bioavailability (AUC=524+/-403 microg min/ml). It was only applied i.v. or i.m. In this work, the influence of absorption enhancers (aggregation and ion-pair formation) on the bioavailability and on the hydrophilic properties of Cp was investigated. The bioavailability of cefpirom was improved through the combination with absorption enhancers (hexadecyldimethylbenzylammonium chloride, BAC; hexylsalicylic acid, HSA). The absolute bioavailability of the Cp combination with absorption enhancers was 21 times larger for BAC and 15 times larger for HSA than in the case when Cp was used alone. PMID- 16377109 TI - 2,3-Dihydroxybenzoic acid attenuates kanamycin-induced volume reduction in mouse utricular type I hair cells. AB - The aminoglycoside kanamycin is a commonly used antibiotic, but unfortunately it is oto- and nephrotoxic in large doses. The negative effects are thought to be due to the formation of free radicals which is why strong antioxidants and iron chelators like 2,3-dihydroxybenzoic acid (DHB) are of great interest. This study estimates cellular quantitative changes in the utricular macula of mice following systemic treatment with kanamycin alone or in combination with DHB. The animals were injected with either saline, kanamycin or kanamycin + DHB for 15 days and perfusion fixed three weeks after last injection. Total volume of the utricle, as well as total number of hair and supporting cells, were estimated on light microscopic sections. Total volume and mean volume of hair cell types I and II and supporting cells were estimated on digital transmission electron micrographs. Total volume of the utricular macula, hair cell type I and supporting cells decreased significantly in animals injected with kanamycin but not in animals co treated with DHB. Hair and supporting cell numbers remained unchanged in all three groups. In conclusion, the kanamycin-induced volume reduction of type I hair cells was attenuated by DHB. PMID- 16377110 TI - Analysis of estrogenic compounds in Polygonum cuspidatum by bioassay and high performance liquid chromatography. AB - The estrogenic activity of traditional Chinese herb-Polygonum cuspidatum Sieb. et Zucc. was investigated by a recombinant yeast screening (YES) assay. Anthraquinones are the main components in the plant, of which emodin is the most abundant one. The ethyl acetate fraction of the ethanol extract of Polygonum cuspidatum was separated on a silica gel TLC plate and seven sub-fractions were collected. The results of bioassay demonstrated that Hzs1 and Hzs6 showed higher estrogenic activities than that of others and the potency of these two compounds were approximately 10(-4) g/L and 10(-3) g/L, respectively. HPLC analysis was performed to determine the activities and the active components. Combining the results of HPLC analysis and estrogenic activity test by YES led to the conclusion that an unknown bioactive compound might exist in the extraction of Polygonum cuspidatum. PMID- 16377111 TI - Aging alters the reduction of pro-apoptotic signaling in response to loading induced hypertrophy. AB - This study tested the hypothesis that loading decreased apoptosis in skeletal muscle in an aging-dependent fashion. One wing of young and aged Japanese quails was loaded for 7 or 21 days to induce hypertrophy. The contralateral wing served as the intra-animal control. Loading increased fast-twitch quail patagialis muscle mass by 28 and 49%, after 7 or 21 days of loading, respectively in young adult birds. Muscle mass was not elevated after 7 days of loading, but increased by 29% after 21 days of loading in aged birds. Seven days of loading reduced DNA fragmentation and cytosolic accumulation of cytochrome c in muscles from young birds but not in muscles from aged birds. ARC protein content was lower and H2O2 content was higher in muscles from aged birds following 7 days of loading. The mitochondria-free cytosolic protein fraction from muscles loaded for 7 days had 41 and 29% lower AIF content than control muscles in young and aged birds, respectively. XIAP, an apoptotic suppressor protein increased after 7 days of loading in muscles from young adult but not aged birds. Our results suggest that loading suppresses pro-apoptotic signaling in quail muscle but aging delays or attenuates these anti-apoptotic changes. PMID- 16377112 TI - Methods of fetal MR: beyond T2-weighted imaging. AB - The present work reviews the basic methods of performing fetal magnetic resonance imaging (MRI). Since fetal MRI differs in many respects from a postnatal study, several factors have to be taken into account to achieve satisfying image quality. Image quality depends on adequate positioning of the pregnant woman in the magnet, use of appropriate coils and the selection of sequences. Ultrafast T2 weighted sequences are regarded as the mainstay of fetal MR-imaging. However, additional sequences, such as T1-weighted images, diffusion-weighted images, echoplanar imaging may provide further information, especially in extra- central nervous system regions of the fetal body. PMID- 16377113 TI - Lung cancer detected in patients presenting to the Emergency Department studies for suspected pulmonary embolism on computed tomography pulmonary angiography. AB - PURPOSE: To study the frequency and demographics of lung cancer on CT pulmonary angiography in patients with suspected pulmonary embolism referred from the Emergency Department. MATERIALS AND METHODS: Retrospective review of the medical records and radiology reports, clinical and imaging follow-up studies and pathological reports revealed 1106 CT pulmonary angiography studies referred from our Emergency Department during the 15-month period between March 2003 and June 2004. RESULTS: Five incidental lung cancer cases were found in 1106 studies from 1081 patients (0.47%). Pulmonary embolism was found in 95 patients (8.5%). Among the five incidental cases three patients were female and two were male (62-81 years old; mean 73 years, 17-130 packs year; mean 51 packs year). Tumor size ranged from 1.8 to 4.5 cm (mean 3.3 cm). The stagings of the lung cancers were IIIB in one patient and IV in four patients. CONCLUSION: Previously undiagnosed lung cancer was detected in 0.45% of patients among 1081 patients referred from Emergency Department, one of whom had coexistent pulmonary embolism. All five patients presented at advanced lung cancer stages of IIIB and IV. PMID- 16377114 TI - Four cases of solid pseudopapillary tumors of pancreas: imaging findings and pathological correlations. AB - OBJECTIVE: Solid pseudopapillary tumor of the pancreas (SPTP tumor) is a rare pancreatic neoplasm with low malignant potential, which usually affects female patients in the second or third decades of life. It is a non-functional, slow growing neoplasm that very often reaches considerable size before the first symptoms appear. Symptomatology is frequently related to tumor size. Surgical excision is usually curative in most cases. Infrequently the tumor can appear in male patients or in aged women, which can make the diagnosis more difficult. Some patients develop liver metastases in the follow-up that can be resected. Our purpose is to review the radiological and pathological findings of SPTP with emphasis on these infrequent cases. SUBJECTS AND METHODS: The medical records and radiological findings of patients who underwent surgery for SPTP between 2000 and 2005 were retrospectively reviewed. Study eligibility required that patients had undergone surgical resection and that a SPTP had been pathologically proved. RESULTS: Four cases of solid pseudopapillary tumor of the pancreas were diagnosed and treated in our institution in the study period. Two of the patients, developed on follow-up liver metastases, and peritoneal, hepatic, and nodal metastases, respectively. CONCLUSION: Solid pseudopapillary tumors are well encapsulated neoplasms that usually have a good prognosis after surgical excision. A malignant behavior is uncommon and in this case lymph node involvement, hepatic metastases and occasionally peritoneal invasion may also occur. Resection of liver metastases can prolong the long-term survival of the patients. PMID- 16377115 TI - Validated HPLC-MS/MS method for simultaneous determination of simvastatin and simvastatin hydroxy acid in human plasma. AB - Cholesterol lowering statin drugs are the most frequently prescribed agents for reducing morbidity and mortality related to coronary heart disease. This publication presents a validated, highly sensitive and selective isocratic HPLC method for the quantitative determination of the major statin drug simvastatin (SIM) and its metabolite simvastatin hydroxy acid (SIMA). Detection was performed on an electrospray ionization triple quadrupole mass spectrometer equipped with an ESI interface operated in positive and negative ionization mode. The multiple reaction-monitoring mode (MRM) was used to provide MS/MS detection. The linearity for the calibration curve in the concentration range of 0.10-16.00 ng/mL for SIM and 0.10-16.00 ng/mL for SIMA is presented. Inter- and intra-day precision and accuracy of the proposed method were characterized by relative standard deviation (R.S.D.) and percentage deviation, respectively; with both lower than 7% for all analytes. The limit of quantitation was 0.03 ng/mL for SIM and 0.02 ng/mL for SIMA. The devised method was employed in the pharmacokinetic study of SIM and the pharmacokinetic parameters of all analytes are also presented. PMID- 16377116 TI - Studies of the pharmacokinetics of paeoniflorin in two Jing-Zhi-Guan-Xin formulations after oral administration to beagle dogs. AB - Paeoniflorin is the principal bioactive component of Paeoniae Radix. The traditional chinese medicine compound recipe (TCMCR) tablets of Jing-Zhi-Guan-Xin (JZGX), which is composed of Radix Salviae Miltiorrhizae, Radix Paeoniae Rubrae, Rhizoma Chuan-xiong, Flos Carthami and Lignum Dalbergiae Odorafera, have been widely used in China and Japan. The plasma concentrations of paeoniflorin in beagle dogs after oral administration of two Jing-Zhi-Guan-Xin formulations (the dose used in the two formulations were both 200 mg paeoniflorin) were measured using a simple and rapid HPLC method. The mean terminal half-lives (t1/2) of JZGX tablet and JZGX elementary osmotic pump tablet (EOPT) formulations of paeoniflorin were 147.52 +/- 28.98 and 276.60 +/- 24.24 min, the maximum plasma concentrations (Cmax) of paeoniflorin were 210.49 +/- 23.89 and 94.36 +/- 14.01 ng/ml, times to reach maximum concentrations (tmax) were 130.00 +/- 30.98 and 280.00 +/- 48.99 min, the area under the plasma concentration-time curves (AUC0 infinity) were 43066.50 +/- 10119.51 and 42266.87 +/- 2654.90 ng min/ml, the mean residence times (MRT) were 212.87 +/- 41.82 and 399.14 +/- 34.98 min, respectively, and the relative bioavailability (Fr) of JZGX EOPT compared with JZGX tablet was 101.8 +/- 18.8%. These results, compared with the pharmacokinetic parameters of paeoniflorin after oral administration of Paeoniae Radix extract alone, indicated that the absorption of paeoniflorin after oral administration of the two JZGX formulations was significantly greater than that after oral administration of Paeoniae Radix extract alone. PMID- 16377117 TI - Investigation of degradation products in a topical gel containing erythromycin and benzoyl peroxide by liquid chromatography-mass spectrometry. AB - Benzamycin, combining benzoyl peroxide and erythromycin, is a topical gel used in the treatment of acne vulgaris. Because of the reactivity of benzoyl peroxide, preparations containing both erythromycin and benzoyl peroxide might be unstable and degradation products could be formed. To investigate and identify these latter products, a gradient-based liquid chromatographic method using volatile mobile phase constituents was developed. Mass spectrometry data were acquired on solutions containing erythromycin and benzoyl peroxide and on freshly prepared, 2 month-old and 18-month-old samples of Benzamycin. With the reference spectra as interpretative templates, it was concluded that erythromycin undergoes oxidation, followed by benzoylation. PMID- 16377118 TI - Flavour retention and release from protein solutions. AB - This paper briefly presents the main results obtained up to now on protein flavour binding and release in relation with flavour perception. Among the food proteins, beta-lactoglobulin is the most extensively studied for its binding properties, which involve both hydrophobic and hydrogen binding. Recent developments using molecular modelling and Quantitative Structure-Activity Relationship confirmed the existence of two different binding sites for flavour compounds on beta-lactoglobulin. During the aroma release process in the mouth, not only free aroma compounds are released but also those reversibly bound by the protein, pointing out the fact that flavour perception is only affected if strong binding occurs. PMID- 16377119 TI - Significance of lymph node micrometastasis in pN0 hilar bile duct carcinoma. AB - AIMS: To examine micrometastasis in node-negative hilar bile duct carcinoma (HBDC) using an immunohistochemical method and evaluated the clinical significance. METHODS: Four hundred and twenty three regional lymph nodes from 28 patients with node-negative HBDC who had undergone a resection were immunostained with an antibody against cytokeratins eight and 18 (CAM 5.2). RESULTS: Lymph node micrometastasis was detected in 11 of the 28 patients and 14 of the 423 lymph nodes. Lymph node micrometastasis was significantly correlated with the pT classification (p=0.03), the histopathological grading (p=0.01) and venous invasion (p=0.05). The 5-year survival rate of the patients with lymph node micrometastasis was 21.8%, as opposed to 66.5% in the patients without micrometastasis. Patients with micrometastasis showed a significantly poorer survival rate than those without micrometastasis (p=0.02). CONCLUSION: The results suggest that immunohistochemically detected lymph node micrometastasis has an impact on the outcome in HBDC. PMID- 16377120 TI - Prognostic factors after surgery for locally recurrent rectal cancer: an overview. AB - PURPOSE: Local recurrence of rectal cancer occurs in a considerable group of patients who have undergone radical treatment for primary tumour. The treatment of choice is surgical resection but the prognosis remains poor, as a negative margin excision is possible in only a small subset of patients. A review of prognostic factors for locally recurrent rectal cancer (LRRC) after surgery is presented. METHODS: We systematically reviewed the literature for reports on prognostic factors after surgical excision of LRRC. These reports were identified through a review of the Medline database from 1982 to 2004. RESULTS: This review highlights the most important prognostic factors for LRRC patients treated with surgery. Data are grouped on the basis of the prognostic factors investigated. CONCLUSIONS: R0 resection seems to be the only reliable prognostic factor; however, symptoms, pre-operative CEA doubling time, performance status and pre operative radiotherapy can help patient selection before surgery. The results of this review provide the basis for improved outcome, aiming to assess patients who would benefit from reoperation. PMID- 16377121 TI - [Recombinant activated factor VII to control bleeding in cardiovascular surgery: need of efficacy and safety validation]. PMID- 16377122 TI - [A rare aetiology of the post-partum fever: ovarian vein thrombophlebitis]. AB - OBJECTIVES: To clarify the contribution of the doppler and the CT in the balance aetiology of a fever of the post-partum and to connect it with a thrombophlebitis of ovarian vein. PATIENTS AND METHODS: Five patients presented there post-partum a fever with pointed abdominal painful syndrome. A doppler and a CT were performed. RESULTS: Doppler showed a hypoechoic tubular structure located forward and laterally with regard to the psoas with a flat spectre in pulsed Doppler in every case. CT confirmed the diagnosis of a thrombophlebitis of the right ovarian vein in three cases and left in two cases. Evolution after anticoagulation and an antibiotic therapy was favourable with regression of clinical signs and doppler evaluation. CONCLUSION: In front of any fever of the post-partum, it is necessary to evoke a thrombophlebitis of the vein ovarian, although it is about a rare aetiology. In spite of the superiority of the CT-scan and RP imaging for such a diagnosis, doppler is a simple and reproducible diagnostic tool for the monitoring which must be practised in first intention. PMID- 16377123 TI - [Guidelines of the French "Haute Autorite de Sante" to improve safety in extracorporeal circulation]. PMID- 16377124 TI - [Hypothermia after spinal anaesthesia: implication of morphine?]. AB - Spinal anaesthesia is the gold standard for elective caesarean section. This technique presents several adverse effects. We report a severe case of hypothermia (33.3 degrees C) after spinal administration of bupivacaine (10 mg) and morphine (100 microg) for elective caesarean section. After excluding other causes of hypothermia, this one could be explained by both the own effects of local anaesthesia (i.e. peripheral vasodilatation) and by the central effect of intrathecal morphine. Because hypothermia is not predictable after spinal injection of morphine both monitoring of central temperature and active warming of the patients could be proposed. Naloxone has been proposed in a case of hypothermia related to spinal injection of morphine. PMID- 16377125 TI - [Prolonged neuromuscular block induced by mivacurium]. PMID- 16377126 TI - Gold-induced reactive oxygen species (ROS) do not mediate suppression of monocytic mitochondrial or secretory function. AB - The toxicity of anti-rheumatic gold compounds has limited their use and development, yet both the toxicological and therapeutic actions of these compounds remain unclear. In the current study, we tested the hypothesis that intracellular reactive oxygen species (ROS) induced by Au(I) or Au(III) compounds mediate their ability to suppress mitochondrial activity. METHODS: Human THP1 monocytes were exposed to HAuCl(4) x 3H(2)O (Au(III)), or the anti-rheumatic compounds auranofin (AF) or gold sodium thiomalate (GSTM) for 6-72 h, after which mitochondrial activity (succinate dehydrogenase) was measured. To assess the role of cellular redox status as a mediator of mitochondrial suppression, monocytes were pre-treated with a pro-oxidant (t-butyl hydroquinone, t-BHQ) or antioxidant (N-acetyl cysteine, NAC ). ROS levels were measured 0-24h post-gold addition to determine their role as mediators of mitochondrial activity suppression. RESULTS: AF was the most potent inhibitor of mitochondrial activity, followed by Au(III) and GSTM. Only Au(III) induced intracellular ROS; no ROS formation was observed in response to AF or GSTM exposure. Although anti- and pro-oxidants had some effects on mitochondrial suppression of Au compounds, collectively the data do not support redox effects or ROS formation as major mediators of Au-compound mitochondrial suppression. CONCLUSIONS: Our results do not indicate that ROS and redox effects play major roles in mediating the cytotoxicity of AF, GSTM or Au(III). PMID- 16377128 TI - Intelligibility of tracheoesophageal speech in noise. AB - The purpose of this investigation is to determine the extent to which background noise negatively impacts the intelligibility of tracheoesophageal (TE) speech. Four male TE speakers provided speech samples that were recorded in quiet and in noise conditions. The listener/subjects occupied a sound-treated booth and were presented with two tasks. In Task 1, the subjects were required to transcribe TE speech stimuli recorded in quiet. In Task 2, the subjects were required to transcribe TE speech stimuli recorded in noise. Repeated measures 2 x 4 factorial analyses of variance were calculated for the dataset. The results of the statistical analysis revealed that the TE speech produced in quiet was significantly more intelligible to the listeners than the TE speech produced in noise for three of the four TE speakers. Furthermore, the results seem to support the hypothesis that the activation of a Lombard effect in TE speakers may detract from their overall speech intelligibility. PMID- 16377127 TI - Rapid evolution of a recently retroposed transcription factor YY2 in mammalian genomes. AB - YY2 was originally identified due to its unusual similarity to the evolutionarily well-conserved zinc finger gene YY1. In this study, we have determined the evolutionary origin and conservation of YY2 using comparative genomic approaches. Our results indicate that YY2 is a retroposed copy of YY1 that has been inserted into another gene locus named Mbtps2 (membrane-bound transcription factor protease site 2). This retroposition is estimated to have occurred after the divergence of placental mammals from other vertebrates based on the detection of YY2 only in the placental mammals. The N- and C-terminal regions of YY2 have evolved under different selection pressures. The N-terminal region has evolved at a very fast pace with very limited functional constraints, whereas the DNA binding, C-terminal region still maintains a sequence structure very similar to that of YY1 and is also well conserved among placental mammals. In situ hybridizations using different adult mouse tissues indicate that mouse YY2 is expressed at relatively low levels in Purkinje and granular cells of cerebellum and in neuronal cells of cerebrum, but at very high levels in testis. The expression levels of YY2 are much lower than those of YY1, but the overall spatial expression patterns are similar to those of Mbtps2, suggesting a possible shared transcriptional control between YY2 and Mbtps2. Taken together, the formation and evolution of YY2 represent a very unusual case where a transcription factor was first retroposed into another gene locus encoding a protease and survived with different selection schemes and expression patterns. PMID- 16377129 TI - Vocal expression of emotions in normally hearing and hearing-impaired infants. AB - The vocalizations of seven normally hearing (NH) and seven severely hearing impaired (HI) infants were compared to find out the influence of auditory feedback on preverbal utterances. It was tested whether there are general differences in vocalizations between NH and HI infants, and whether specific emotional states affect the vocal production of NH and HI infants in the same way. First, the acoustic structure of the three most common vocal types was analyzed; second, the composition of vocal sequences was examined. Vocal sequence composition turned out to be more affected by hearing impairment than the acoustic structure of single vocalizations. This result indicates that the acoustic structure of preverbal vocalizations is to a great extent predetermined, whereas the composition of vocal sequences is influenced by auditory input. PMID- 16377130 TI - Phonatory impairment in Parkinson's disease: evidence from nonlinear dynamic analysis and perturbation analysis. AB - Many persons with Parkinson's disease (PD) will eventually experience vocal impairment as their condition advances. Using standard perturbation analyses (parameters like jitter and shimmer) to measure fluctuations in phonatory signal may inhibit researchers from recognizing severely disordered patterns that seem to be present in the voices of some PD patients. Nonlinear dynamic analysis can quantify these aperiodic patterns, which indicate severe pathology that is usually characterized perceptually by hoarseness. Here, sustained vowel phonations of a heterogeneous group of PD subjects (20 women and 21 men) were compared with those of a control group (22 women and 18 men) based on results of nonlinear dynamic analyses (D(2)) and perturbation analyses. Results showed PD subjects as a whole to have significantly higher D(2) values than control subjects (P = 0.016), which indicates increased signal complexity in PD vocal pathology. Differences in the comparison of these two groups were significant in jitter (P = 0.014) but nonsignificant in shimmer (P = 0.695). Furthermore, the performance on these three measures was affected by subject sex. Nonlinear dynamic analysis showed significantly higher D(2) in the female PD group than in the female control group (P = 0.001), but jitter and shimmer did not show such a difference. The male PD group had statistically higher jitter than the male control group (P = 0.036), but these groups did not differ in D(2) or shimmer. Overall, nonlinear dynamic analysis may be a valuable method for the diagnosis of Parkinsonian laryngeal pathology. PMID- 16377131 TI - Accuracy evaluation of image registration and segmentation tools used in conformal treatment planning of prostate cancer. AB - Segmentation and registration tools are commonly used in radiotherapy for target and at risk organs localisation. In this work the performances of three different segmentation tools and of a surface matching registration technique, used on computed tomography (CT) and magnetic resonance (MR) images for the treatment planning of conformal prostate carcinoma, are studied. The accuracy of the segmentation and registration tools was evaluated by phantom experiment and on patient data, respectively. A preliminary estimate of MR image distortion was also performed. PMID- 16377132 TI - Phosphorylation of the Pro-X-Thr-Pro site in phosphatase inhibitor-2 by cyclin dependent protein kinase during M-phase of the cell cycle. AB - Protein phosphorylation serves as a primary mechanism for triggering events during mitosis and depends on coordinated regulation of kinases and phosphatases. Protein Ser-Thr phosphatase-1 (PP1) activity is essential for the metaphase to anaphase transition and the most ancient regulator of PP1 conserved from yeast to human is inhibitor-2 (I-2), an unstructured heat-stable protein. A unique sequence motif in I-2 from various species surrounds a phosphorylation site PXTP that can be phosphorylated in biochemical assays by GSK3, MAPK and CDK kinases. Here we used a phosphosite specific antibody to investigate the phosphorylation of I-2. We fractioned extracts from HeLa cells arrested with nocodazole and assayed for PXTP kinases using recombinant I-2. One major and two minor peaks of kinase activity were identified and the major peak contained both active MAPK and cdk1::cyclinB1, confirmed by immunoblotting. Cells released from a double thymidine block synchronously progressed through mitosis and immunoblotting revealed transient phosphorylation of endogenous I-2 in cells only during mitosis, and corresponding phosphorylation of histone H3 (Ser10) and PP1 (Thr320). Activation of cdk1::cyclinB1 was coincident with I-2 phosphorylation, but neither MAPK nor GSK3 were phosphorylated at this time, so we concluded that in living cells only cdk1::cyclinB1 phosphorylated the PXTP site in I-2. Immunofluorescent staining of cells with the PXTP phosphosite antibody revealed highly specific staining of mitotic cells prior to anaphase, at which point the staining disappeared. Thus, phosphorylation of I-2 is catalyzed by cdk1::cyclinB1 and staining with a specific antibody should prove useful as a selective marker of cells in the early stages of mitosis. PMID- 16377133 TI - Pontine activation during focal status epilepticus secondary to hamartoma of the floor of the fourth ventricle. AB - Epileptic seizures associated with hamartoma of the floor of the fourth ventricle (HFFV) are generally resistant to antiepileptic medication, may evolve into status epilepticus, and can respond favorably to surgical therapy. HFFV are rare, and during the neonatal or infantile period may be associated with repetitive and stereotyped attacks of hemifacial spasm, eye blinking, facial movements, head deviation and dysautonomic manifestations. Similarly, to gelastic seizures provoked by hypothalamic hamartomas, it has been suggested that these spells arise from within the HFFV, thus constituting a type of non-cortical seizure. We report an infant female patient that developed continuous left hemifacial attacks since she was 2-month-old, and that underwent presurgical investigation when she was 18-month-old. MRI disclosed a left sided HFFV, Video-EEG showed non localizing and non-lateralizing findings, and SPECT aligned with MRI showed marked hyperperfusion within the hamartoma, spreading to ipsilateral cerebellar parenchyma and brainstem nuclei. Patient underwent lesionectomy and became seizure-free. We found two evidences on literature supporting the hypothesis of non-cortical seizures related to HFFV. The first, intra-cerebellar recordings surrounding hamartoma showed electrical activity related to seizures. The second, subtracted SPECT co-registered MRI showed hyperemia within hamartoma. The present report provides the third additional evidence. We found the involvement not only of the hamartoma, and pars of cerebellar hemisphere, but also an intense hyperemia involving brainstem nuclei during seizures. We believe that all these findings suggest a short subcortical network responsible for generating seizures in HFFV patients. PMID- 16377134 TI - Antiepileptic drug trials in the elderly. AB - It is important to perform clinical trials of antiepileptic drugs (AEDs) in elderly because they differ in many ways from younger people. The response of elderly people to AEDs may not be predictable from studies focused exclusively on younger adults. Major issues of concern include altered absorption and metabolism, as well as the likelihood of interactions with concomitant medications. PMID- 16377135 TI - Obtaining pediatric indications for new anti-epileptic drugs: how and when. AB - Drug development in children poses a number of challenges that must be overcome to obtain adequate information in product labeling. Trials in children must be supported by appropriate toxicology and formulation work, which tends to delay completion of work in children until after the drug is available for adults. The Pediatric Research Equity Act (PREA) contains a decision tree to help companies devise an appropriate pediatric program for drugs in development. The medical community does not currently endorse the assumption that the progression of epilepsy and response to treatment is the same in adults and children. Therefore, a complete drug development program in children is necessary and includes efficacy and safety trials along with pharmacokinetic studies. These studies are needed to justify the risk/benefit in children. Formulations appropriate for children are needed. Seizure diaries must be maintained by caretakers and in the case of infants, seizures may need to be counted by EEG. Early planning and discussion of a pediatric program with regulatory agencies will facilitate this work. PMID- 16377136 TI - Models of epileptogenesis in adult animals available for antiepileptogenesis drug screening. AB - Epileptogenesis is the process by which parts of a normal brain are converted to a hyperexcitable brain, often after an injury. Researchers must understand this process before they know where and how to change it. Animal models are used to evaluate the process of epileptogenesis by studing status epelepticus, electrical kindling, or other methods that provoke injuries. All are associated with neuronal loss to more or less degree, synaptic reorganization, axon sprouting, neurogenesis, gliosis, and changes in gene expression in neurons and astrocytes. He describes several types of animal models and how they might be useful in developing effective strategies for preventing epilepsy. PMID- 16377137 TI - Antiepileptic drug trials - VIII. An overview of issues in the development of new antiepileptic drugs. Symposium proceedings. Key Biscayne, Florida, USA, March 17 19, 2005. PMID- 16377138 TI - Adverse psychiatric effects of antiepileptic drugs. AB - Psychiatric symptoms provide potential insights into the pathophysiology of psychiatric conditions in epilepsy patients. Evaluating the psychiatric effects of newer AEDs is complicated by several important factors including (a) the relatively high rate of mood and anxiety disorders in persons with pharmacoresistant epilepsy, and (b) the variety of methods used to assess the occurrence and severity of psychiatric symptoms. Clinical trials to assess adverse psychiatric effects of AEDs need a control group to adjust for the spontaneous occurrence of symptoms, as well as to demonstrate potential positive psychotropic effects. The follow-up should be long enough to differentiate between drug-related effects and the natural course of the comorbid disorder. Psychiatric problems should be defined using appropriate instruments as well as self-report. PMID- 16377139 TI - Large clinical trials in epilepsy: funding by the NIH versus pharmaceutical industry. AB - Different approaches to clinical trials are taken by government-sponsored (NIH, VA, MRC) and industry-sponsored clinical trials. Each sector has a different perspective and funding capacity. Sponsorship often depends on the question. Initial safety and efficacy studies for regulatory purposes usually are industry driven. Studies of drug use as monotherapy, direct comparisons among drugs, and comparisons between categories of drugs often are more important to prescribers and payors, thereby requiring government funding. The challenge to clinician investigators is to now find ways to fund comparison trials to determine evidence based guidelines so patients receive the best treatment both in the short- and long-term, and the health care system gets the best value. PMID- 16377140 TI - Antiepileptic drugs trials: neonates and infants. AB - Special issues are related to AED testing in several populations. Pharmacokinetics, pharmacodynamics and underlying neurochemistry and developing systems require specific testing in appropriate infants with refractory seizures. EEG monitoring is an essential part of seizure definition and recognition, making it a necesssity along with clinical semiology to define the seizure types and changes in seizure frequency. (1) Neonates: A trial design for neonatal seizures should be similar to those used for the treatment of status epilepticus. Proposed study end points should be seizure cessation for some period of time, or time to next seizure. The use of placebo is questionable. (2) Partial seizures occurring in young infants: A trial designed for topirimate included a placebo controlled, double blind study with fixed dose trials exploring the range of tolerated doses. Forty-eight hour video EEGs were used for quantification of seizures. (3) Uncommon forms of encephalopathic epilepsy: a proposed design includes randomization to sequential monotherapy with prescribed titration/dose defined by seizure control or tolerance. Outcome variables include seizure reduction, tolerability, and time continued on AED. PMID- 16377141 TI - Clinical trials for prevention of epileptogenesis. AB - Methods to evaluate treatments that could prevent epileptogenesis in subjects at risk are described. Epilepsy prevention trials are more complex, lengthy, and costly than standard epilepsy treatment trials for many reasons. Issues revolve around selection of subjects, consent for participation, length of follow-up, and selection of an appropriate endpoint. The use of biomarkers are a possible solution in future research. PMID- 16377142 TI - Pediatric epilepsy models. AB - Pediatric epilepsy models are needed to help with development of drugs for specific childhood and infantile epilepsy syndromes. The major forms of pediatric epilepsy can be divided into those that occur in the neonatal period, infancy, early childhood and late childhood. Seizures in the immature brain are different from those in adult brain, often resulting in neuronal death. Rodent models are useful in mimicking seizures in the immature brain (neonatal seizures, infantile spasms, and febrile seizures). No specific models exist for syndromes (e.g., Lennox-Gastaut, Landau-Kleffner). The interaction between brain development and epilepsy in humans can be assisted by use of high resolution MRI, diffusion tensor imaging, and functional MRI. PMID- 16377143 TI - FDA update. AB - This report addresses: (1) a general update of FDA activity in areas relevant to AED development; (2) an update on issues relevant to the development of AEDs in the pediatric population; and (3) an update on the Agency's approach to the evaluation of AEDs as monotherapy. FDA ACTIONS: Since January 2002, 47 Approval actions for 10 AEDs were issued, but none for a new chemical entity. Nine of the ten Approvable actions taken were relatively minor changes to existing applications. An Approvable letter was issued for Lyrica (pregabalin) for the treatment of post-herpetic neuralgia, painful diabetic neuropathy, and partial seizures in adults. The primary issue to be addressed in the face of post marketing reports of adverse events is one of causality. The FDA has requested that sponsors search their databases for selected problems under review (e.g., suicidality). PEDIATRICS: The Pediatric Research Equity Act (PREA) and the Best Pharmaceuticals for Children Act (BPCA) require studies in pediatric patients for those indications granted for adults that are relevant for the pediatric population. Current FDA policy asks sponsors to undertake a development program in pediatric patients essentially analogous to that for adults. MONOTHERAPY TRIALS: Establishing the effectiveness of AEDs as monotherapy continues to be desirable, but problematic. Problems include the difficulty of performing monotherapy trials, ethical issues, designation of patients as "newly diagnosed," and endpoints. Historical controls may be acceptable if: (a) there is a consensus that it is essentially impossible to conduct controlled trials designed to demonstrate a difference between treatments; (b) there is an adequate historical database against which the seizure rate seen with the new drug can reasonably be compared; and (c) there is evidence from adequate and well-controlled trials that the treatment is effective as adjunctive therapy. FDA is Agency is reviewing analyses describing historical controls. PMID- 16377144 TI - Bioequivalence in development of antiepileptic drugs. AB - Bioequivalence is an important component of the development of AEDs. Development of new formulations after the original testing of any drug requires demonstration that the compounds are therapeutically equivalent and additional efficacy studies may not be required. Extended-release formulations may reduce toxicity with a lower maximum blood concentration (C(max)) and improve efficacy with a higher minimum blood concentration (C(min)). Obtaining an equivalent area under the curve (AUC) while slowing the gastrointestinal transit and avoiding food effects and dose dumping among a population with epilepsy with individual variability requires extensive engineering of the formulation. The development of extended release divalproex (Depakote ER) is used as an example of the challenges of this phase of drug development. Other routes of administration discussed are rectal preparations, nasal formulations, and intravenous infusions. These newer formulations may offer better patient care and more efficient development. PMID- 16377145 TI - Historical data in the design and interpretation of trials with newly diagnosed patients. AB - An International League Against Epilepsy (ILAE) subcommission is exploring the possibility of utilizing historical data from treated and control (untreated or under-treated) patients who have been enrolled in monotherapy trials in newly diagnosed epilepsy. Active-control trials have not convincingly distinguished between equivalent effectiveness and equivalent ineffectiveness. Thus, there is insufficient historical evidence of sensitivity to drug effects. Noninferiority trials are based upon experience acquired with previous trials. Optimizing the exploitation of historical data obtained in newly diagnosed patients may be a way to improve the validity of future trials. A concern about use of historical controls is lack of assurance that historical populations and newly recruited populations are similar. The best approach to pooling and modeling data may be an Individual Patient Data (IPD) approach as (1) analysis of data from active control trials, (2) demonstration that the subpopulation carrying an intermediate probability of recurrence is the most sensitive to drug effect, by comparison to similar subpopulations who received a placebo or a pseudoplacebo in historical studies. This subpopulation could become a reference for patient selection or stratification in future trials to improve assay sensitivity. PMID- 16377146 TI - Historical control withdrawal to monotherapy. AB - The design of clinical trials can be explained in terms of selection of a control group. Two options have included (a) using an active comparator, usually a standard drug such as carbamazepine (active control) or (b) using a placebo or low-dose control. Because neither active control nor placebo/dose control appeared to be acceptable alternatives for monotherapy studies in epilepsy, a new concept has been considered: the use of "historical control". This method uses the "expected" (imputed) behavior of placebo, based on prior trials. Some trials have been performed in patients with refractory epilepsy, who are withdrawn from their previous antiepileptic drugs, and converted to monotherapy on a study drug, using a low-dose comparator, frequently called "pseudo-placebo." A meta-analysis of all pseudo-placebo arms was performed to determine whether they could serve as a historical control for future monotherapy trials. The estimate of the combined percent exit was 86.1% (CI 78.6% to 93.6%). These data may be able to serve as a historical control for future monotherapy studies, obviating the need for a placebo/pseudo placebo arm. PMID- 16377147 TI - Active control comparisons: the ideal trial design. AB - This report describes the concept for a clinical trial that uses carbamazepine as the gold-standard active control for a study of newly diagnosed patients. The authors describe an endpoint including efficacy and tolerability, and a stopping rule that uses a series of interim analyses in order to reach a conclusion as efficiently as possible without sacrificing reliability. PMID- 16377148 TI - Cognitive and memory effects of the new antiepileptic drugs. AB - Problems with cognition are common in patients with epilepsy. A series of double blind, randomized, crossover, healthy volunteer studies have been conducted to avoid a variety of the confounding effects on cognition such as those produced by changes in seizure frequency. All of the older AEDs produced cognitive effects compared to the non-drug conditions. The cognitive effects of several of the new AEDs are described although data are limited. Studies have demonstrated that in utero AED exposure in humans may affect cognitive development. Although the cognitive effects of AEDs are generally modest, these effects can have clinical significance. The available data suggest that some of the new AEDs have fewer effects on cognition and memory than the older AEDs, and these differences can have clinical impact. PMID- 16377149 TI - Therapeutic drug monitoring assay development to improve efficacy and safety. AB - The development of therapeutic drug management (TDM) utilizing diagnostic assays as biomarker tests is described. TDM can be useful in establishing an individual patient's optimal blood concentration range, and benchmarking blood concentrations at which seizures are controlled, as well as those associated with AED-specific adverse effects. TDM requires the application of pharmacokinetic, pharmacodynamic, pharmacogenomic, pathophysiological and clinical principles to the management of patients in order to achieve safe and effective therapy. Optimal therapy requires rational application of all these principles to personalize patient care. PMID- 16377150 TI - How trial designs impact on guidelines. AB - Evidence-based treatment guidelines and practice parameters have been developed in most areas of medicine. They are important because insurance companies, formulary managers, and health maintenance organizations rely on guidelines for formulary and coverage decisions. The methodology for practice parameter development by the AAN is described as it was applied to evaluating AEDs. Systematic criteria are used to classify the evidence as class I, II, III or IV. There is value in designing clinical trials so they will be considered to provide a high level of evidence. The major variables that will be assessed include presence of randomization, use of a control group, use of masked outcome assessment, and in an active-controlled trial, use of an adequate comparator and adequate enrollment to detect a difference if one exists. Minor variables also must be addressed, including a priori definition of the primary outcome variable, clearly defined Inclusion/exclusion criteria, presence of equivalent treatment groups at baseline, adequate duration of assessment to answer the clinical question, and appropriate management and statistical handling of drop-outs. Understanding of the variables that will be considered for treatment guidelines can improve the methodological strength of studies and lead to recommendations for practice. PMID- 16377151 TI - Molecular targets versus models for new antiepileptic drug discovery. AB - Animal models have played a key role in the discovery and characterization of all marketed antiepileptic drugs (AED). The conventional wisdom is that the standard animal screening models are becoming obsolete because they fail to identify compounds that act in mechanistically new ways and as a result do not offer therapeutic advantages over presently available agents. In fact, far from only detecting me-too drugs, the models often uncover compounds with distinctive profiles of activity in various types of epilepsy and in addition have unexpected efficacy in non-epilepsy conditions, such as neuropathic pain, bipolar disorder, and migraine. Moreover, the animal models-because they are unbiased with respect to mechanism-provide an opportunity to uncover drugs that act in new ways and through new targets, such as alpha2delta and SV2A. In vitro testing is not likely to replace screening in animal models because in vitro systems cannot model the specific pharmacodynamic actions required for seizure protection, and do not assess bioavailability and brain accessibility. PMID- 16377152 TI - Active control trials: endpoints beyond conventional efficacy and tolerability measures. AB - Clinical trial endpoints often extend beyond conventional efficacy and tolerability measures. While it is generally assumed that seizure control without major adverse drug effects is by far the most important determinant of quality of life, other factors also are important. Pharmacokinetic endpoints could also provide relevant information for optimal drug use. Outcome measures could reveal drug effects on mood, headache, and sleep disorders, as well as overall quality of life and seizure severity. PMID- 16377153 TI - Proof of Principle studies. AB - Proof of Principle studies are an early stage of clinical drug development when a compound has shown potential in animal models and early safety testing. This step of proof-of-principle (PoP) or proof-of-concept (PoC) often links between Phase-I and dose ranging Phase-II studies. These small-scale studies are designed to detect a signal that the drug is active on a pathophysiologically relevant mechanism, as well as preliminary evidence of efficacy in a clinically relevant endpoint. Sponsors use these studies to estimate whether their compound might have clinically significant efficacy in other diseases states as well as epilepsy (e.g., migraine, neuropathic pain, anxiety, depression). Endpoints other than seizure frequency often reveal special characteristics of the drug. A structured dose escalation design can reveal dose-dependent effects and adverse effects, crossover studies can demonstrate change, presurgical studies can define efficacy, interictal discharges and photosensitivity models can explain changes in seizure features, transcranial magnetic stimulation evaluates hyperexcitability. PoP studies allow exploration of a wide range of potential therapeutic areas beyond epilepsy as part of an integrated CNS development plan. PMID- 16377154 TI - Increased medial temporal lobe activation during the passive viewing of emotional and neutral facial expressions in schizophrenia. AB - INTRODUCTION: Patients with schizophrenia show deficits in facial affect and facial identity recognition and exhibit structural and neurophysiological abnormalities in brain regions known to mediate these processes. Functional neuroimaging studies of neural responses to emotional facial expressions in schizophrenia have reported both increases and decreases in medial temporal lobe (MTL) activity in schizophrenia. Some of this variability may be related to the tasks performed and the baseline conditions used. Here we tested whether MTL responses to human faces in schizophrenia are abnormal when unconstrained by a cognitive task and measured relative to a low-level baseline (fixation) condition. METHODS: 15 patients with schizophrenia and 16 healthy control subjects underwent functional magnetic resonance imaging (fMRI) while passively viewing human faces displaying fearful, happy, and neutral emotional expressions. RESULTS: Relative to control subjects, the patients demonstrated (1) significantly greater activation of the left hippocampus while viewing all three facial expressions and (2) increased right amygdala activation during the initial presentation of fearful and neutral facial expressions. CONCLUSIONS: In schizophrenia, hippocampal and amygdala activity is elevated during the passive viewing of human faces. PMID- 16377155 TI - Self-initiated encoding facilitates object working memory in schizophrenia: implications for the etiology of working memory deficit. AB - BACKGROUND: Working memory (WM) deficit is present in a majority of patients with schizophrenia but it is unclear which components of WM are impaired. Past studies suggest that encoding may be compromised. One important determinant of encoding is the deployment of selective attention to the target stimulus. In addition, attention and encoding are modulated by motivational factors. In this study, we investigated the effects of self-initiated encoding (i.e., voluntary attention) on WM. METHODS: 19 patients with schizophrenia and 19 matched control subjects participated in visual WM and control tasks. Encoding was manipulated by asking subjects to select from two face targets and memorize 1) one of the two identical faces (Non-preference condition), 2) one that is marked (Non-choice condition), and 3) one they prefer (Preference condition). WM accuracy for both location (spatial) and identity (object) was measured. RESULTS: Overall, patients with schizophrenia were less accurate and slower than the control subjects but the deficit was greater for object WM. However, patients were more accurate in object WM when they selected a preferred face as their target during encoding (preference condition) compared with the other two conditions. This effect was not significant for spatial WM. CONCLUSIONS: These results suggest that voluntary, self-initiated attention may facilitate object encoding especially if the selection of the target involves affective choice, and that attention may play different roles in encoding 'what' versus 'where' in WM. Since encoding affects all forms of memory, these results may have a more general implication for memory. PMID- 16377156 TI - Brain morphology in first-episode schizophrenia: a meta-analysis of quantitative magnetic resonance imaging studies. AB - BACKGROUND: A number of meta-analytic reviews of structural brain imaging studies have shown that multiple subtle brain abnormalities are consistently found in schizophrenia. However, quantitative reviews till now published have included mainly studies performed on chronic schizophrenic patients but have failed to provide clear information on specific, possibly different, findings in first episode schizophrenia. METHODS: We performed a systematic search for MRI studies that reported quantitative measurements of volumes of brain regions in first episode schizophrenic patients and in healthy controls. Twelve meta-analyses were performed for 6 cerebral regions. RESULTS: Twenty-one studies were identified as suitable for analysis. Significant overall effect sizes were demonstrated for lateral and third ventricular volume increase, and for volume reduction of whole brain and hippocampus, but not for temporal lobe, amygdala and total intracranial volumes. CONCLUSIONS: The available literature data strongly indicate that some brain abnormalities are already present in first-episode schizophrenic patients. However, unlike the results of published meta-analyses conducted primarily on samples of chronic schizophrenic patients, the present study did not confirm a significant reduction of temporal lobe or amygdala volumes in first-episode schizophrenia. These findings support the hypothesis of different patterns of involvement of various cerebral areas over the time course of schizophrenia. PMID- 16377157 TI - The detection of neurocognitive decline in schizophrenia using the Mindstreams Computerized Cognitive Test Battery. AB - BACKGROUND: The Mindstreams Computerized Cognitive Test Battery (Mindstreams) is a standardized computer-based battery that was designed for widespread clinical and research use. The capability of Mindstreams to test cognitive impairment in schizophrenia has yet to be evaluated. The aim of the present study was to determine the ability of Mindstreams in detecting cognitive dysfunction in schizophrenia patients and to compare it to the Cambridge Neuropsychological Test Automated Battery (CANTAB). METHOD: Fifty-five schizophrenia patients and 63 healthy subjects were enrolled in the study. The Positive and Negative Syndrome Scale (PANSS) was used to quantify symptom severity. Neurocognitive functions were assessed using Mindstreams and CANTAB. RESULTS: The schizophrenia patients scored significantly more poorly than healthy subjects on all tests comprising the Mindstreams battery. Comparable tasks of the Mindstreams and CANTAB batteries significantly correlated on raw scores and the standardized cognitive indices. The Mindstreams executive function tasks had significant correlations with the PANSS negative, autistic preoccupation and activation cluster scores, and with global functioning. Two-week test-retest reliability correlations were all significant (N=17, p<0.05-p<0.001). CONCLUSIONS: This study indicates that Mindstreams is reliable in assessing the cognitive function of patients with schizophrenia and may play a role in standardizing the cognitive assessment of these patients in clinical and research settings. PMID- 16377158 TI - Clinical significance of sleep EEG abnormalities in chronic schizophrenia. AB - This study aimed to investigate the relationship between measures of clinical symptom severity and sleep EEG parameters in a relatively diagnostically homogeneous group of patients with schizophrenia. We obtained sleep EEG data in 15 drug-free inpatients who met DSM-IV-R criteria for schizophrenia, undifferentiated type, with 15 age- and sex-matched normal controls over two consecutive night polysomnographic recordings. Clinical symptoms were assessed by the Positive and Negative Symptom Scale (PANSS) and Hamilton Rating Scale for Depression. Characteristic features of sleep disturbance were seen in patients with schizophrenia: profound difficulties in sleep initiation and maintenance, poor sleep efficiency, a slow wave sleep (SWS) deficit, and an increased REM density. SWS was inversely correlated with cognitive symptoms. REM density was inversely correlated with positive, cognitive, and emotional discomfort symptoms as well as PANSS total score. Our data demonstrate that drug-free patients with chronic undifferentiated type schizophrenia suffer from profound disturbances in sleep continuity and sleep architecture. Both the SWS deficit and cognitive impairment found in schizophrenics in this study may relate to similar underlying structural brain abnormalities. PMID- 16377159 TI - Screening for Neuroligin 4 (NLGN4) truncating and transmembrane domain mutations in schizophrenia. PMID- 16377160 TI - A novel method for automated EMG decomposition and MUAP classification. AB - OBJECTIVE: This paper proposes a novel method for the extraction and classification of individual motor unit action potentials (MUAPs) from intramuscular electromyographic signals. METHODOLOGY: The proposed method automatically detects the number of template MUAP clusters and classifies them into normal, neuropathic or myopathic. It consists of three steps: (i) preprocessing of electromyogram (EMG) recordings, (ii) MUAP detection and clustering and (iii) MUAP classification. RESULTS: The approach has been validated using a dataset of EMG recordings and an annotated collection of MUAPs. The correct identification rate for MUAP clustering is 93, 95 and 92% for normal, myopathic and neuropathic, respectively. Ninety-one percent of the superimposed MUAPs were correctly identified. The obtained accuracy for MUAP classification is about 86%. CONCLUSION: The proposed method, apart from efficient EMG decomposition addresses automatic MUAP classification to neuropathic, myopathic or normal classes directly from raw EMG signals. PMID- 16377161 TI - Spray coated pellets as carrier system for mucoadhesive drug nanocrystals. AB - High pressure homogenization can be employed to produce drug nanocrystals with a number of advantages, like improved solubility behaviors, better drug targeting or even increased mucoadhesiveness. To obtain a controlled drug delivery system it is necessary to transform the resulting nanosuspension into a solid dosage form. The present study shows the feasibility to use a mucoadhesive nanosuspension of poorly soluble hydrocortisone acetate produced by high pressure homogenization as layering dispersion in a fluidized bed process, followed by the application of an enteric coating to achieve a controlled drug release. To point out the advantages of drug nanocrystals the new fomulation was compared with a formulation containing micronized drug. Both formulations were characterized with regard to their particle size and crystallinity by using laser diffractometry, photon correlation spectroscopy and X-ray diffraction. The pellet morphology was characterized by using the environmental scanning electron microscopy (ESEM). In the in vitro dissolution tests an accelerated dissolution velocity and an increased drug release could be shown for the pellets containing drug nanocrystals. PMID- 16377162 TI - Differential segmental growth of the vertebral column of the rat (Rattus norvegicus). AB - Despite the pervasive occurrence of segmental morphologies in the animal kingdom, the study of segmental growth is almost entirely lacking, but may have significant implications for understanding the development of these organisms. We investigate the segmental and regional growth of the entire vertebral column of the rat (Rattus norvegicus) by fitting a Gompertz curve to length and age data for each vertebra and each vertebral region. Regional lengths are calculated by summing constituent vertebral lengths and intervertebral space lengths for cervical, thoracic, lumbar, sacral, and caudal regions. Gompertz curves allow for the estimation of parameters representing neonatal and adult vertebral and regional lengths, as well as initial growth rate and the rate of exponential growth decay. Findings demonstrate differences between neonatal and adult rats in terms of relative vertebral lengths, and differential growth rates between sequential vertebrae and vertebral regions. Specifically, relative differences in the length of vertebrae indicate increasing differences caudad. Vertebral length in neonates increases from the atlas to the middle of the thoracic series and decreases in length caudad, while adult vertebral lengths tend to increase caudad. There is also a general trend of increasing vertebral and regional initial growth and rate of growth decay caudad. Anteroposterior patterns of growth are sexually dimorphic, with males having longer vertebrae than females at any given age. Differences are more pronounced (a) increasingly caudad along the body axis, and (b) in adulthood than in neonates. Elucidated patterns of growth are influenced by a combination of developmental, functional, and genetic factors. PMID- 16377163 TI - Intramembranous ossification of scleral ossicles in Chelydra serpentina. AB - Scleral ossicles are present in many reptiles, including turtles and birds. In both groups the sclerotic ring situated in the eye is composed of a number of imbricating scleral ossicles or plates. Despite this gross morphological similarity, Andrews (1996. An endochondral rather than a dermal origin for scleral ossicles in Cryptodiran turtles. J. Herpetol. 30, 257-260) reported that the scleral ossicles of turtles develop endochondrally unlike those in birds, which develop intramembranously after a complex epithelial-mesenchymal inductive event. This study re-explores one of the species examined by Andrews in order to determine the mode of ossification of scleral ossicles in turtles. A growth series of Chelydra serpentina embryos, including the stages examined by Andrews, were examined by staining separately for cartilage and bone. Results clearly contradict Andrews (1996) and show that the scleral ossicles of Chelydra serpentina develop similarly to those in birds. That is, they develop intramembranously without a cartilage precursor and are likely induced by transient scleral papillae. The sequence of scleral papillae development is broadly similar, but the papillae themselves are not as distinct as those seen in chicken embryos. This study has important consequences for understanding the homology of scleral ossicles among tetrapods. PMID- 16377164 TI - Cross-sectional geometry of the dentary in bats. AB - Bats exhibit remarkable diversity in dietary habits, with species specializing on insects, fruit, nectar, vertebrates and blood. Studies of larger mammals have shown that structural differences in dentary cross-sectional properties exist among species with different diets. Unfortunately, few of these studies have considered the role of phylogeny in shaping these apparent form-function associations. Here we ask whether a relationship exists between diet and dentary structure in bats when phylogenetic history is factored into the analysis. To answer this question, we compared results from phylogenetic generalized least squares (PGLS) and traditional (nonphylogenetic) regression analyses of dentary cross-sectional shape in frugivorous, nectarivorous, and insectivorous bats (253 individuals representing 72 species). Cross-sectional moments of inertia of the dentary between M(1) and M(2) were computed from bone densitometry scans of skeletal specimens. Traditional regressions of cross-sectional parameters against dentary length detected significant departures from isometry among frugivores. In contrast, PGLS analyses indicated that cross-sectional variables for each dietary group scaled with isometry. Thus, the allometric patterns illuminated by traditional statistics are linked to the phylogenetic structure of the sample. Identical patterns of significant differences in slopes and intercepts between frugivores and nectarivores emerged from both traditional and PGLS analyses. As predicted, the cross-sectional shape of the dentary in frugivores is consistent with increased resistance to torsion and bending, while that of nectarivores suggested a less resistant dentary. Although traditional and PGLS analyses yielded some similar results, the phylogenetic structure of a sample can drive apparent patterns of scaling and should be considered in comparative functional analyses. PMID- 16377165 TI - Chitinolytic activities of ericoid mycorrhizal and other root-associated fungi from Epacris pulchella (Ericaceae). AB - Ericoid mycorrhizal endophytes and other root-associated fungi from Epacris pulchella (Ericaceae) in an eastern Australian sclerophyll forest, along with Hymenoscyphus ericae, were tested for their abilities to produce extracellular chitinolytic activities during growth in axenic culture. Two root-associated fungi produced activities that were active against only a monomeric 4 methylumbelliferyl (4-MU) glycoside of N-acetylglucosamine, suggesting exo-acting beta-N-acetylhexosaminidase (EC 3.2.1.52) activity. All ericoid mycorrhizal fungi and two root-associated fungi produced activities against dimeric and trimeric 4 MU glycosides of N-acetylglucosamine, suggesting production of chitobiosidase and endo-acting chitinase (EC 3.2.1.14) respectively in addition to beta-N acetylhexosaminidase. Specific activities for all ericoid mycorrhizal fungi, including H. ericae, were of the same order of magnitude, suggesting that their chitinolytic potential is broadly similar. Chitinase activities were only produced by an ericoid mycorrhizal fungus when chitin was included in the medium, however, no activity was produced if glucose was also present in the medium. PMID- 16377166 TI - Genetic diversity of the Chestnut blight fungus Cryphonectria parasitica in four French populations assessed by microsatellite markers. AB - Microsatellites are powerful markers to infer population genetic parameters. Here, 13 microsatellite loci isolated from a genomic and a cDNA library of Cryphonectria parasitica were used to characterize the genetic diversity and structure of four French populations. Twelve of these loci were polymorphic within populations, and average gene diversity (H(e)) was estimated to be 0.35. There was a lower genetic diversity in a south-eastern population relative to three south-western populations. In these three populations, microsatellite genotypic diversity was higher than vegetative compatibility type diversity. A high genetic differentiation (G(ST) = 0.27) suggested a low gene flow and/or founder effects of French populations which are in agreement with low dispersal of spores and different introductions of this species in southern France. This study demonstrates the significance of these microsatellite loci to assess gene flow and reproductive system in this important pathogen. PMID- 16377168 TI - A GIS based transportation model for solid waste disposal--a case study on Asansol municipality. AB - Uncontrolled growth of the urban population in developing countries in recent years has made solid waste management an important issue. Very often, a substantial amount of total expenditures is spent on the collection of solid waste by city authorities. Optimization of the routing system for collection and transport of solid waste thus constitutes an important component of an effective solid waste management system. This paper describes an attempt to design and develop an appropriate storage, collection and disposal plan for the Asansol Municipality Corporation (AMC) of West Bengal State (India). A GIS optimal routing model is proposed to determine the minimum cost/distance efficient collection paths for transporting the solid wastes to the landfill. The model uses information on population density, waste generation capacity, road network and the types of road, storage bins and collection vehicles, etc. The proposed model can be used as a decision support tool by municipal authorities for efficient management of the daily operations for transporting solid wastes, load balancing within vehicles, managing fuel consumption and generating work schedules for the workers and vehicles. The total cost of the proposed collection systems is estimated to be around 80 million rupees for the fixed cost of storage bins, collection vehicles and a sanitary landfill and around 8.4 million rupees for the annual operating cost of crews, vehicles and landfill maintenance. A substantial amount (25 million rupees/yr) is currently being spent by AMC on waste collection alone without any proper storage/collection system and sanitary landfill. Over a projected period of 15 yr, the overall savings is thus very significant. PMID- 16377169 TI - Conferences on solid waste management. PMID- 16377170 TI - Linking microbial community structure with function: fluorescence in situ hybridization-microautoradiography and isotope arrays. AB - The ecophysiology of microorganisms has been at the heart of microbial ecology since its early days, but only during the past decade have methods become available for cultivation-independent, direct identification of microorganisms in complex communities and for the simultaneous investigation of their activity and substrate uptake patterns. The combination of fluorescence in situ hybridization (FISH) and microautoradiography (MAR) is currently the most widely applied tool for revealing physiological properties of microorganisms in their natural environment with single-cell resolution. For example, this technique has been used in wastewater treatment and marine systems to describe the functional properties of newly discovered species, and to identify microorganisms responsible for key physiological processes. Recently, the scope of FISH-MAR was extended by rendering it quantitative and by combining it with microelectrode measurements or stable isotope probing. Isotope arrays have also been developed that exploit the parallel detection offered by DNA microarrays to measure incorporation of labelled substrate into the rRNA of many community members in a single experiment. PMID- 16377171 TI - Stem cells in the etiology and treatment of cancer. AB - Using approaches first applied in human leukemias, recent progress has been made in the identification of putative cancer stem cells in several different carcinomas and other solid cancers. Additional studies have suggested that cancer stem cells may be derived not only from transformation of quiescent, long-term stem cells but also from short-lived progenitors that then obtain the ability to undergo self-renewal. Therefore, the heterogeneity observed in many types of human cancers may reflect the activation of specific oncogenes and/or loss of specific tumor suppressor genes and the different stem and/or progenitor cell populations in which these genetic or epigenetic events occur. Similarities have been observed in the pathways regulating stem cell homing and metastasis, and increasing evidence also suggests that treatment failure and the recurrence of human cancer may reflect the intrinsic quiescence and drug resistance of cancer stem cells. PMID- 16377172 TI - Matrix metalloproteinase-induced genomic instability. AB - Increased expression of matrix metalloproteinases (MMPs) is associated with nearly every tumor type. Although many studies have shown that MMPs can promote malignancy, recent evidence has revealed that MMPs can play a causative role also in the earliest stages of cancer development. A complex story is now emerging in which MMPs not only compromise cell-cell and cell-substratum adhesion processes that impact genomic surveillance mechanisms but also act directly on molecules at the cell surface to stimulate physiological processes that cause genetic alterations. Delineating the mechanisms involved in these processes and identifying how they are coordinated in vivo could aid identification of the crucial contribution of MMPs to tumorigenesis. PMID- 16377173 TI - Invasion is a genetic program regulated by transcription factors. AB - The invasive and metastatic behaviour of tumours impacts crucially on the clinical management of cancer. Accordingly, it is important to understand the regulation of tumour cell invasiveness. Genetic analysis of worms, Drosophila and mice has provided evidence that invasion is a genetic pathway regulated by transcription factors that are often implicated in tumour cell invasion. Recent evidence has revealed much concerning the role of one particular transcription factor, AP1, which is involved in the regulation of a multigenic invasion program in which upregulated and downregulated genes function as invasion effectors and suppressors, respectively. Differentially expressed genes cooperatively enhance pseudopod elongation during the mesenchymal mode of invasion by altering the function, localisation and activity of non-differentially expressed proteins. PMID- 16377174 TI - Balancing cell adhesion and Wnt signaling, the key role of beta-catenin. AB - Controlled regulation of cell proliferation and differentiation is essential for embryonic development and requires the coordinated regulation of cell-cell adhesion and gene transcription. The armadillo repeat protein beta-catenin is an important integrator of both processes. Beta-catenin acts in the Wnt signaling pathway, activating the transcription of crucial target genes responsible for cellular proliferation and differentiation. Beta-catenin also controls E-cadherin mediated cell adhesion at the plasma membrane and mediates the interplay of adherens junction molecules with the actin cytoskeleton. Both functions of beta catenin are de-regulated in human malignancies, thereby leading both to the loss of cell-cell adhesion and to the increased transcription of Wnt target genes. PMID- 16377175 TI - Inhibition of TGFbeta signaling in cancer therapy. AB - Recent evidence continues to support a central role for TGFbeta in tumor maintenance and progression. Although this may involve TGFbeta-mediated paracrine effects that modulate the tumor microenvironment and the host immune system, some studies causally implicate autocrine TGFbeta in cancer cell motility and survival. Other recent evidence indicates synergy between oncogene and TGFbeta signaling in epithelial cell transformation. This suggests opportunities for dissecting molecular mechanisms of cross-talk as well as providing insights into possible combinatorial molecular anticancer therapies that will include TGFbeta inhibitors. PMID- 16377176 TI - Specific peptides for the therapeutic targeting of oncogenes. AB - Tumors are dependent on oncogenic proteins for their maintenance and survival. The ideal cancer therapy would include drugs that specifically target these proteins. Many such proteins function through interfaces that can be difficult to target effectively with small molecules. However, recent advances in cell permeable peptide technology, improving cellular penetration and stability, raise the possibility that specific peptide interference of oncogenic proteins could be successfully translated to the clinic. Several active anti-tumor peptides were recently described. For example, a stable peptide inhibitor of the Hsp90 ATP binding pocket killed a wide range of tumors in vitro and in vivo, and a peptide inhibitor of the BCL6 oncoprotein was active in B-cell lymphomas; both peptides functioned without toxicity to normal tissues. PMID- 16377177 TI - Development of wiring specificity in the olfactory system. AB - The olfactory system discriminates a large number of odorants using precisely wired neural circuits. It offers an excellent opportunity to study mechanisms of neuronal wiring specificity at the single synapse level. Each olfactory receptor neuron typically expresses only one olfactory receptor from many receptor genes (1000 in mice). In mice, this striking singularity appears to be ensured by a negative feedback mechanism. Olfactory receptor neurons expressing the same receptor converge their axons to stereotypical positions with high precision, a feature that is conserved from insects to mammals. Several molecules have recently been identified that control this process, including olfactory receptors themselves in mice. The second order neurons, mitral cells in mammals and projection neurons in insects, have a similar degree of wiring specificity: studies in Drosophila suggest that projection neuron-intrinsic mechanisms regulate their precise dendritic targeting. Finally, recent studies have revealed interactions of different cell types during circuit assembly, including axon-axon interactions among olfactory receptor neurons and dendro-dendritic interactions of projection neurons, that are essential in establishing wiring specificity of the olfactory circuit. PMID- 16377178 TI - Clamp loaders and replication initiation. AB - Clamp loaders are ATP-driven multiprotein machines that couple ATP hydrolysis to the opening and closing of a circular protein ring around DNA. This ring-shaped clamp slides along DNA, and interacts with numerous proteins involved in DNA replication, DNA repair and cell cycle control. Recently determined structures of clamp loader complexes from prokaryotic and eukaryotic sources have revealed exciting new details of how these complex AAA+ machines perform this essential clamp loading function. PMID- 16377179 TI - Quality of life in long-term Hodgkin's disease survivors with chronic fatigue. AB - The prevalence of chronic fatigue (CF), elevated fatigue levels >six months, is 2.5-3 times higher in long-term Hodgkin disease survivors (HDSs) than in the general population (GP). This is the first study comparing the quality of life (QoL) of chronic fatigued survivors with that of chronic fatigued subjects in the GP. The fatigue questionnaire (FQ) and the SF-36 were mailed to 591 HDSs successfully treated at the Norwegian Radium Hospital from 1971 to 1997. Patient data were compared with nationally representative GP data, adjusted for age, gender and education. The response rate was 80% (N = 475), median follow-up 195 months (range 53-431), mean age 46 years (range 21-74), and 44% were females. HDSs reported significantly poorer QoL than the GP with lower scores on six of eight SF-36 scales (P < 0.001). The subgroup of HDSs with chronic fatigue however (N = 142/475, 30%) had better QoL on five of eight SF-36 scales than subjects with CF in the GP (N = 224/2141, 11%), particularly mental health and emotional role-functioning (P = 0.005 and <0.0001). No differences were found on the physical SF-36 domains. Overall, HDSs reported lower QoL than the GP, in particular the HDSs with CF. Their QoL was similar to that of GP subjects with CF, but with significantly better mental health status. This suggests that CF in long-term survivors may be associated with more physical than psychological aspects of long-term cancer survivorship. PMID- 16377180 TI - Prognostic value of lymphovascular invasion in women with lymph node negative invasive breast carcinoma. AB - This study aimed to test the hypothesis that lymphovascular invasion adds prognostic information to histological grade and tumour size in node-negative invasive carcinoma of the breast. Lymphovascular invasion was assessed in haematoxylin and eosin tumour sections from 2760 patients with node-negative invasive breast carcinoma treated with definitive surgery. Patients were divided into two groups: 990 in the no adjuvant therapy series (diagnosed in 1974-1988) with median follow-up of 13 years; and 1765 in the selective adjuvant therapy series (1988-2000) with median follow-up of 6.8 years. Lymphovascular invasion was identified in 19% of tumours and was associated with larger tumour size, higher histological grade and younger age. Overall, survival was associated on multivariate analysis with lymphovascular invasion, histological grade and tumour size in both patient series, and with histological type in the no adjuvant therapy series. In conclusion, lymphovascular invasion is an independent prognostic factor in node-negative breast cancer and should be considered in decisions about adjuvant treatment in this group of women. PMID- 16377181 TI - The experiences of men with breast cancer in the United Kingdom. AB - To investigate the experiences of men with breast cancer across the United Kingdom, a multi-phase study using: (a) focus groups (n = 4) with men and women with breast cancer and with healthcare professionals; (b) questionnaires to men with breast cancer (n = 161); (c) follow-up interviews with these men (n = 30) and (d) reconvening the focus groups (n = 2) for the men and women with breast cancer. The majority of men (84%, n = 135) reported their symptoms early, but were shocked to receive a breast cancer diagnosis. Disclosure of the diagnosis was commonly made to partners (80%, n = 129) and other close family and was influenced by perceptions of embarrassment, stigma and altered body image. Very little information was available to participants; that which was available was often inappropriate as it was intended for women. Over half the sample wanted much more information (56%, n = 90). This study also demonstrated low utilisation of formal support services and initiatives are needed to improve the information and support provided to men with breast cancer after diagnosis and treatment. Increasing the profile of breast cancer in men generally amongst healthcare professionals and the public is also needed. PMID- 16377182 TI - DNA triplex stabilization by a delta-carboline derivative tethered to third strand oligonucleotides. AB - A delta-carboline derivative was covalently coupled to a 7 mer oligonucleotide at its 5'- or 3'-end. The stability of triplexes formed from the conjugates and a double-helical target was studied by UV melting experiments. Compared to the unmodified control triple helices, triplexes with the conjugate exhibit a significantly higher stability. However, the degree of stabilization depends on the particular triplex structure formed. PMID- 16377183 TI - New photoantimicrobial films composed of porphyrinated lipophilic cellulose esters. AB - Porphyrinated cellulose laurate esters have been prepared in homogeneous DMA/LiCl medium by "one-pot, two-step" reactions starting from cellulose, protoporphyrin IX, and lauric acid and using a TsCl/Pyridine system. The plastic films obtained after casting were shown to display photobactericidal activity against Gram positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacteria. This new photobactericidal polymer has potential for industrial, medical, or household applications. PMID- 16377184 TI - Natural and synthetic G-quadruplex interactive berberine derivatives. AB - The interaction of the natural alkaloid berberine with various G-quadruplex DNA structures and its ability to inhibit telomerase have been examined and compared with those of a synthetic piperidino derivative and the related compound coralyne. The results show that these molecules have selectivity for G-quadruplex compared to duplex DNA, and that their aromatic moieties play a dominant role in quadruplex binding. PMID- 16377185 TI - Synthesis of the first sulfur-35-labeled hERG radioligand. AB - The synthesis of the first high specific activity S-35-labeled hERG radioligand, [(35)S]MK-0499, for use in HTS assays of drug candidates for hERG interaction is described. The radioligand is prepared by [(35)S]sulfonylation of a high diastereomeric excess (de) aniline precursor prepared from unlabeled MK-0499. PMID- 16377186 TI - 1,4-Anhydrogalactopyranose is not an intermediate of the mutase catalyzed UDP galactopyranose/furanose interconversion. AB - UDP-galactopyranose mutase (UGM) catalyzes the isomerization of UDP galactopyranose (UDP-Galp) into UDP-galactofuranose (UDP-Galf), an essential step of the mycobacterial cell wall biosynthesis. The first mechanistic assumption proposed in the literature was the involvement of 1,4-anhydrogalactose 1 as intermediate of this ring contraction. To confirm or rule out this hypothesis, we synthesized 1 and engaged it in reactions with UGM. The expected formations of UDP-Galf and UDP-Galp were never observed, thus showing that 1 is not, in fact, a low energy intermediate of this enzymatic contraction. PMID- 16377187 TI - Oxadiazole derivatives as a novel class of antimitotic agents: Synthesis, inhibition of tubulin polymerization, and activity in tumor cell lines. AB - Oxadiazole derivatives were synthesized and evaluated for their ability to inhibit tubulin polymerization and to cause mitotic arrest in tumor cells. The most potent compounds inhibited tubulin polymerization at concentrations below 1 microM. Lead analogs caused mitotic arrest of A431 human epidermoid cells and cells derived from multi-drug resistant tumors (10, EC(50)=7.8 nM). Competition for the colchicine binding site and pharmacokinetic properties of selected potent compounds were also investigated and are reported herein, along with structure activity relationships for this novel series of antimitotic agents. PMID- 16377190 TI - Characteristics of patients who did not wait for treatment in the emergency department: a follow up survey. AB - A significant number of patients leave the emergency department (ED) before being treated or after treatment has been initiated but not completed. This paper reports the findings of a study examining the demographics and characteristics of those who did not wait for treatment in an ED in Melbourne. A telephone survey was undertaken to examine the reasons they did not wait. Data were collected in four one-month periods across the year and patients were telephoned within 72 h of their departure from the ED. The majority of those who did not wait were Australian Triage Scale category 4 or 5, male, and with a mean age of 37 years. Sixty per cent of those who did not wait presented between the hours of 6 pm and 6 am and waited an average of 130 min before leaving. Over fifty per cent (127) of those surveyed (n=243) sought treatment elsewhere and over a third (72) thought their problem was inappropriate for an ED. However, the need for communication with patients in the waiting room should not be under-played. PMID- 16377188 TI - Novel endotoxin-sequestering compounds with terephthalaldehyde-bis guanylhydrazone scaffolds. AB - We have shown that lipopolyamines bind to the lipid A moiety of lipopolysaccharide, a constituent of Gram-negative bacterial membranes, and neutralize its toxicity in animal models of endotoxic shock. In an effort to identify non-polyamine scaffolds with similar endotoxin-recognizing features, we had observed an unusually high frequency of hits containing guanylhydrazone scaffolds in high-throughput screens. We now describe the syntheses and preliminary structure-activity relationships in a homologous series of bis guanylhydrazone compounds decorated with hydrophobic functionalities. These first generation compounds bind and neutralize lipopolysaccharide with a potency comparable to that of polymyxin B, a peptide antibiotic known to sequester LPS. PMID- 16377191 TI - Breach avoidance facilitator--managing the A&E 4-hour target. AB - This paper aims to explore the concept of the breach avoidance facilitator (BAF) within the Accident & Emergency (A&E) department. This is a role that has been introduced utilising Senior Nursing staff on a supernumerary basis to predominantly provide a trouble-shooting role to manage the A&E 4-hour target. The paper will explore the initial practical difficulties that were encountered during the introduction of the role. The associated benefits of the role including monitoring of the 4-hour target, co-ordination of resources, increased communication with a variety of staff members, and the completion of a real time electronic hand-over form will be explored. The paper will also discuss how the role will evolve to provide support and clinical supervision for junior and ancillary staff. PMID- 16377192 TI - A moral dilemma in the emergency room: confidentiality and domestic violence. AB - A professional colleague presents for treatment at an Emergency Department. Investigations reveal signs that indicate the possibility of domestic violence. This is subsequently confirmed in confidence. Personal reflections identify and explore the dilemmas and the ethical issues involved. PMID- 16377193 TI - 3D-QSAR and docking studies of aldehyde inhibitors of human cathepsin K. AB - In order to better understand the structural and chemical features of human cathepsin K (CatK), which is an important cysteine protease in the pathogenesis of osteoporosis, the 3D-QSAR (CoMFA) studies were conducted on recently explored aldehyde compounds with known CatK inhibitory activities. The genetic algorithm of GOLD2.2 has been employed to position 59 aldehyde compounds into the active sites of CatK to determine the probable binding conformation. Good correlations between the predicted binding free energies and the experimental inhibitory activities suggested that the identified binding conformations of these potential inhibitors are reliable. The docking results also provided a reliable conformational alignment scheme for 3D-QSAR model. Based on the docking conformations, highly predictive comparative molecular field analysis (CoMFA) was performed with q2 value of 0.723. The predictive ability was validated by some compounds that were not included in the training set. Furthermore, the CoMFA model was mapped back to the binding sites of CatK, to get a better understanding of vital interactions between the aldehyde compounds and the protease. The CoMFA field distributions are in good agreement with the structural characteristics of the binding groove of the CatK, which suggested that the n-Bu in R4 position is the favor group substitute at P1 and moderate groups in R2 group are required on P2 substitute. In addition, 3D-QSAR results also demonstrated that aldehyde is an important pharmacophore because of electrostatic effect. These results, together with the good correlations between the inhibitory activities and the binding free energies predicted by GOLD2.2, demonstrated the power of combining docking/QSAR approach to explore the probable binding conformations of compounds at the active sites of the protein target, and further provided useful information in understanding the structural and chemical features of CatK in designing and finding new potential inhibitors. PMID- 16377194 TI - Synthesis and in vitro evaluation of pseudosaccharinamine derivatives as potential elastase inhibitors. AB - Pseudosaccharinamine derivatives were evaluated for elastase inhibitory activity. Ester derivatives of pseudosaccharinamine displayed reversible and high inhibition of human leukocyte elastase (HLE) as compared to porcine pancreatic elastase (PPE). Cyanomethyl (2S,3S)-2-(1,1-dioxobenzo[d]isothiazol-3-ylamino)-3 methylpentanoate was found to inhibit HLE at Ki=0.8 microM. PMID- 16377195 TI - Synthesis, antiproliferative and antifungal activities of some 2-(2,4 dihydroxyphenyl)-4H-3,1-benzothiazines. AB - A new method for the synthesis of 4H-3,1-benzothiazine skeleton is described. The compounds were obtained by the reaction of sulfinylbis(2,4-dihydroxythiobenzoyl) with o-substituted anilines bearing an activated methylene group (-CH2OH, CH2NR1R2), o-aminobenzanilides or 2-aminobenzophenones. The reaction proceeded through thiobenzanilide intermediates, which were converted to the 4H-3,1 benzothiazine fused ring by an endocyclization process. The compounds were tested for their antiproliferative properties against the cells of a human breast cancer T47D line. The activity of some compounds was comparable to that of cisplatin, studied as a control. A strong antifungal effect against the strains of moulds, yeasts and dermatophytes was also found. PMID- 16377196 TI - Synthesis and pharmacology of pyrido[2,3-d]pyrimidinediones bearing polar substituents as adenosine receptor antagonists. AB - Amino-substituted pyrido[2,3-d]pyrimidinediones have previously been found to bind to adenosine A1 and A2A receptors in micromolar concentrations. The present study was aimed at studying the structure-activity relationships of this class of compounds in more detail. Most of the investigated compounds were provided with polar substituents, such as ethoxycarbonyl groups and basic amino functions, in order to improve their water-solubility. The compounds were synthesized starting from 6-amino-1,3-dimethyluracil via different reaction sequences involving (cyano)acetylation, Vilsmeier formylation, or reaction with diethyl ethoxymethylenemalonate (EMME). The most potent and selective compound of the present series was 6-carbethoxy-1,2,3,4-tetrahydro-1,3-dimethyl-5-(2 naphthylmethyl)aminopyrido[2,3-d]pyrimidine-2,4-dione (11c) with a Ki value of 5 nM at rat and 25 nM at human A1 receptors. The compound was more than 60-fold selective versus A3 and more than 300-fold selective versus A2A receptors. It showed an over 300-fold improvement with respect to the lead compound. In GTPgammaS binding studies at membranes of Chinese hamster ovary cells recombinantly expressing the human adenosine A1 receptor, 11c behaved as an antagonist with inverse agonistic activity. A regioisomer of 11c, 6-carbethoxy 1,2,3,4-tetrahydro-1,3-dimethyl-7-(2- naphthylmethyl)aminopyrido[2,3-d]pyrimidine 2,4-dione (7a) in which the 2-naphthylmethylamino substituent at position 5 of 11c was moved to the 7-position, was a relatively potent (Ki=226 nM) and selective (>20-fold) A3 ligand. In the series of compounds lacking an electron withdrawing ethoxycarbonyl or cyano substituent in the 6-position, compounds with high affinity for adenosine A2A receptors were identified, such as 1,2,3,4 tetrahydro-1,3-dimethyl-5-(1-naphthyl)aminopyrido[2,3-d]pyrimidine-2,4-dione 16b (Ki human A2A=81.3 nM, Ki human A1=153 nM, and Ki human A3>10,000 nM). PMID- 16377197 TI - Synthesis and antifungal activity of substituted-10-methyl-1,2,3,4 tetrahydropyrazino[1,2-a]indoles. AB - Series of substituted-10-methyl-1,2,3,4-tetrahydropyrazino[1,2-a]indoles derivatives have been synthesized and examined for their activity against pathogenic strains of Aspergillus fumigatus (ITCC 4517), Aspergillus flavus (ITCC 5192) Aspergillus niger (ITCC 5405) and Candida albicans (ITCC No 4718). All synthesized compounds showed mild to moderate activity, except for 2-substituted 10-methyl-1,2,3,4-tetrahydropyrazino[1,2-a]indoles 6a-d. The most active 1-(4 chlorophenyl)-10-methyl-1,2,3,4-tetrahydropyrazino[1,2-a]indole 4c exhibited a MIC value of 5.85 microg/disc against A. fumigatus and 11.71 microg/disc against A. flavus and A. niger in disc diffusion assay. Anti-Aspergillus activity of active compound 4c by microbroth dilution assay was found to be 15.62 microg/ml in case of A. fumigatus and 31.25 microg/ml with A. flavus and A. niger. The MIC90 value of the most active compound by percent germination inhibition assay was found to be 15.62 microg/ml against A. fumigatus. The MIC90 values of substituted-10-methyl-1,2,3,4-tetrahydropyrazino[1,2-a]indoles against C. albicans ranged from 15.62 to 250 microg/ml. The in vitro toxicity of the most active 1-(4-chlorophenyl)-10-methyl-1,2,3,4-tetrahydropyrazino[1,2-a]indole 4c was evaluated using haemolytic assay, in which the compound was found to be non toxic to human erythrocytes up to a concentration of 312.50 microg/ml. The standard drug amphotericin B exhibited 100% lysis at a concentration of 37.5 microg/ml. PMID- 16377198 TI - Rapid and efficient oxidation of Hantzsch 1,4-dihydropyridines with sodium periodate catalyzed by manganese (III) Schiff base complexes. AB - Rapid and efficient oxidation of Hantzsch 1,4-dihydropyridine with sodium periodate is reported. The Mn(III)-salophen/NaIO4 catalytic system converts 1,4 dihydropyridines to their corresponding pyridine derivatives at room temperature in a 1:1, CH3CN/H2O mixture. The ability of various Schiff base complexes in the oxidation of 1,4-dihydropyridine was also investigated. PMID- 16377199 TI - Synthesis of verbenachalcone congeners and their biological assessment against activation of the NGF-mediated neurite outgrowth of PC12D cells' activity. AB - Synthesis of the verbenachalcone derivatives 3-5 involving littorachalcone 2 from diaryl ether 7 enabled an SAR study of enhancement activity against the NGF mediated neurite outgrowth from PC12D cells. Littorachalcone 2 and o deoxyverbenachalcone 5 showed similar activity to that of verbenachalcone 1. PMID- 16377200 TI - QSAR models for Daphnia toxicity of pesticides based on combinations of topological parameters of molecular structures. AB - A topological parameter is defined as an integer value of a given local or global invariant of a molecular graph. We examined three types of local graph invariants, the vertex degrees (0EC), the extended connectivity of first order (1EC), and the numbers of paths of length two (P2), as elementary invariants for construction of quantitative structure-activity relationships (QSAR). We also examined combined invariants, obtained by multiplying one of these three elementary types with another (i.e., [0EC.1EC], [0EC.P2], and [1EC.P2]), as graph invariants. Finally, global invariants were used in the QSAR analyses, codifying the presence and nature of cycles in the molecular structures under consideration. We used the correlation weights of these invariants to obtain optimal descriptors. These descriptors have been used in one-variable models to predict toxicity toward Daphnia magna for a set of pesticides. Statistical characteristics of the best model, based on the correlation weight of local topological parameters (the [0EC.P2]) together with the global topological parameters, are the following: n=220, r2=0.7822, s=0.849, F=783 (training set); n=42, r2=0.7388, s=0.941, F=113 (test set). The role of these topological parameters is discussed. PMID- 16377201 TI - Synthesis, biological evaluation, and pharmacokinetic study of prolyl-1 piperazinylacetic acid and prolyl-4-piperidinylacetic acid derivatives as VLA-4 antagonists. AB - A series of prolyl-1-piperazinylacetic acid and prolyl-4-piperidinylacetic acid derivatives were synthesized and evaluated for their activity as VLA-4 antagonists. Of 22 compounds synthesized, 19 compounds showed potent activity with low nanomolar IC50 values. In addition, the representative compounds 11o and 11p with a hydroxy group in the pyrrolidine ring showed moderate plasma clearance in rats (11o, 30 ml/min/kg and 11p, 21 ml/min/kg) and in dogs (11o, 12 ml/min/kg and 11p, 9 ml/min/kg). PMID- 16377202 TI - An overview of copper radionuclides and production of 61Cu by proton irradiation of (nat)Zn at a medical cyclotron. AB - In this article, production methods and applications of copper radionuclides are overviewed with special attention toward (61)Cu, due to its interesting nuclear properties. Selection of production parameters for (61)Cu including: appropriate nuclear reaction, proton beam energy, target thickness and targetry method are discussed for NRCAM 30MeV medical cyclotron. (64)Zn(p,alpha)(61)Cu was selected as the best reaction and (61)Cu was produced by 22MeV proton bombardment of a 80 microm thick natural zinc target. After 180microAh irradiation, the resultant activity of (61)Cu was 6.006Ci (12.015mCi/microAh). The chemical separation method was easy, quick and efficient (>95%) and yielded a no carrier added product with high chemical and radionuclidic purity (>99%). Detailed comparison with previous production methods confirms that our results are superior to other reports published to date. PMID- 16377203 TI - Cytokine interactions in mesenchymal stem cells from cord blood. AB - We used cytokine protein array to analyze the expression of cytokines from human cord blood-derived mesenchymal stem cells (CB-MSCs). Several cytokines, interleukins (IL), and growth factors, including ENA-78, GM-CSF, GRO, IL-1beta, IL-6, IL-8, MCP-1, OSM, VEGF, FGF-4, FGF-7, FGF-9, GCP-2, IGFBP-1, IGFBP-2, IGFBP 3, IGFBP-4, IP-10, LIF, MIF, MIP-3alpha, osteoprotegerin, PARC, PIGF, TGF-beta2, TGF-beta3, TIMP-1, as well as TIMP-2, were secreted by CB-MSCs, while IL-4, IL-5, IL-7, IL-13, TGF-beta1, TNF-alpha, and TNF-beta were not expressed under normal growth conditions. IL-6, IL-8, TIMP-1, and TIMP-2 were the most abundant interleukins expressed by CB-MSCs. A set of growth factors were selected to evaluate their stimulatory effects on the IL6 secretion for CB-MSCs. IL-1beta was the most important factor inducing CB-MSC to secret IL-6. The mechanism by which IL-1beta promoted IL-6 expression in CB-MSCs was studied. By using various inhibitors of signal transduction, we found that activation of p38 mitogen activated protein kinases (MAPK) and MAPK kinase (MEK) is essential in the IL 1beta stimulated signaling cascade which leads to the increase in IL-6 synthesis. Additionally, continuous supplement of IL-1beta in the CB-MSCs culture will facilitate adipogenic maturation of CB-MSCs as evidenced by the presence of oil drops in the CB-MSCs and secretion of leptin, a molecule marker of adipocytes. These results strongly suggest that cytokine induction and signal transduction are important for the differentiation of CB-MSCs. PMID- 16377204 TI - High efficiency single step production of expression plasmids from cDNA clones using the Flexi Vector cloning system. AB - The success of structural genomics and proteomics initiatives is dependent on the availability of target genes in vectors suitable for protein production. Here, we compare two high-throughput methods for producing expression vectors from plasmid derived cDNA fragments. Expression vectors were constructed for compatibility with the Gateway recombination cloning system and the Flexi Vector restriction based cloning system. Cloning protocols for each system were conducted in parallel for 96 different target genes from PCR through the production of sequence-verified expression clones. The short nucleotide sequences required to prepare the target open reading frames for Flexi Vector cloning allowed a single step PCR protocol, resulting in fewer mutations relative to the Gateway protocol. Furthermore, through initial cloning of the target open reading frames directly into an expression vector, the Flexi Vector system gave time and cost savings compared to the protocol required for the Gateway system. Within the Flexi Vector system, genes were transferred between four different expression vectors. The efficiency of gene transfer between Flexi Vectors depended on including a region of sequence identity adjacent to one of the restriction sites. With the proper construction in the flanking sequence of the vector, gene transfer efficiencies of 95-98% were demonstrated. PMID- 16377206 TI - Two-dimensional crystallization and analysis of projection images of intact Thermus thermophilus V-ATPase. AB - H(+)-ATPase/synthases are membrane-bound rotary nanomotors that are essential for energy conversion in nearly all life forms. A member of the family of the vacuolar-type ATPases (V-ATPases) from Thermus thermophilus, sometimes also termed A-type ATPase, was purified to homogeneity and subjected to two dimensional (2D) crystallization trials. A novel approach to the 2D crystallization of unstable complexes yielded densely packed sheets of V-ATPase, exhibiting crystalline arrays. Aggregation of the V-ATPase under acidic conditions during reconstitution circumvented the continuous dissociation of the whole complex into the V(1) and V(o) domains. The resulting three-dimensional aggregates were converted into 2D sheets by the use of a basic buffer, and after a short annealing cycle, ordered arrays of up to 1.5 microm diameter appeared. Fourier transforms calculated from micrographs taken from the negatively stained sample showed diffraction spots to a resolution of 23A. The Fourier transforms of the untilted images revealed unit-cell dimensions of a=232A, b=132A, and gamma=90 degrees , and a projection map was calculated by merging 11 images. The most probable molecular packing suggests p22(1)2(1) symmetry of the crystals and dimer contacts between the V(1) domains. PMID- 16377205 TI - Assessing methods for identifying pair-wise atomic contacts across binding interfaces. AB - An essential step in understanding the molecular basis of protein-protein interactions is the accurate identification of inter-protein contacts. We evaluate a number of common methods used in analyzing protein-protein interfaces: a Voronoi polyhedra-based approach, changes in solvent accessible surface area (DeltaSASA) and various radial cutoffs (closest atom, Cbeta, and centroid). First, we compared the Voronoi polyhedra-based analysis to the DeltaSASA and show that using Voronoi polyhedra finds knob-in-hole contacts. To assess the accuracy between the Voronoi polyhedra-based approach and the various radial cutoff methods, two sets of data were used: a small set of 75 experimental mutants and a larger one of 592 structures of protein-protein interfaces. In an assessment using the small set, the Voronoi polyhedra-based methods, a solvent accessible surface area method, and the closest atom radial method identified 100% of the direct contacts defined by mutagenesis data, but only the Voronoi polyhedra-based method found no false positives. The other radial methods were not able to find all of the direct contacts even using a cutoff of 9A. With the larger set of structures, we compared the overall number contacts using the Voronoi polyhedra based method as a standard. All the radial methods using a 6-A cutoff identified more interactions, but these putative contacts included many false positives as well as missed many false negatives. While radial cutoffs are quicker to calculate as well as to implement, this result highlights why radial cutoff methods do not have the proper resolution to detail the non-homogeneous packing within protein interfaces, and suggests an inappropriate bias in pair-wise contact potentials. Of the radial cutoff methods, using the closest atom approach exhibits the best approximation to the more intensive Voronoi calculation. Our version of the Voronoi polyhedra-based method QContacts is available at . PMID- 16377207 TI - Crystallization and preliminary X-ray crystallographic analysis of the N-terminal photosensory module of phytochrome Agp1, a biliverdin-binding photoreceptor from Agrobacterium tumefaciens. AB - Phytochromes are photochromic photoreceptors with a bilin chromophore that have been found in plants and bacteria. Typical bacterial phytochromes are composed of an N-terminal photosensory chromophore module and a C-terminal protein kinase. The former contains the chromophore, which allows phytochromes to adopt the two interconvertible spectral forms, Pr and Pfr. The N-terminal photosensory module of Agrobacterium phytochrome Agp1, Agp1-M15, was used for crystallization studies. The protein was either assembled with the natural chromophore biliverdin or a sterically locked synthetic biliverdin-derivative, termed 15Za. The last named adduct does not undergo photoisomerization due to an additional carbon chain between the rings C and D of the chromophore. Both adducts could be crystallized, but the resolution was largely improved by the use of 15Za. Crystals of biliverdin-Agp1-M15 diffract to 6A resolution and belong to the tetragonal space group I422 with unit cell dimensions a = b = 171 Angstroms, c = 81 Angstroms, crystals of 15Za-Agp1-M15 belong to the same space group with similar unit cell dimensions a = b = 174 Angstroms, c = 80 Angstroms, but diffract to 3.4 Angstroms resolution. Assuming the asymmetric unit to be occupied by one monomer of 55kDa, the unit cell contains 54-55% solvent with a crystal volume per protein mass, V(m), of 2.7 Angstroms(3) Da(-1). PMID- 16377208 TI - Production of reactive oxygen species (ROS) by devil stinger (Inimicus japonicus) during embryogenesis. AB - In aqua-cultural industry, the seed production of devil stinger, a valuable fish in Japan, has not succeeded yet due to the cryptogenic mass mortality. We found that survival rate of the larvae of devil stinger increased by the addition of green tea extract rich in catechin into rearing tank. Generation of reactive oxygen species (ROS) was detected in the embryo of devil stinger by chemiluminescence analysis under the normal growth conditions without addition of specific stimulants. Even in the unfertilized egg, certain level of ROS was detected. ROS were continuously detected during the development from fertilized egg to larva and tended to increase gradually. Observation of embryos and post hatching larvae with hypersensitive photon-counting microscopy indicated that ROS were produced on the surface of embryo and the head region of larva especially peripheries of eyes. When the embryo proteins were analyzed by immunoblotting using antibody against the human neutrophil cytochrome b558 large subunit (gp91 phox), a main band of approximately 91 kDa was detected, suggesting the presence of NADPH oxidase-like ROS generating system in the embryo of devil stinger. After treatment with streptomycin and penicillin G for 1 day, the level of ROS production in larvae decreased with increase in the survival rate of larvae. Our results suggest that devil stinger has ROS generation system that is already activated at fairly early stage of development before the maturation of usual immune system. PMID- 16377209 TI - Differential expression of three Paralichthys olivaceus Hsp40 genes in responses to virus infection and heat shock. AB - Heat shock proteins (Hsps) are a family of highly conserved cellular proteins present in all organisms, mediating a range of essential housekeeping and cytoprotective functions as well-known molecular chaperones and recently as regulators of the immune response. By subtractive suppression hybridization, three Hsp40 homologues have been identified in the flounder (Paralichthys olivaceus) embryonic cells (FEC) after treatment with UV-inactivated turbot (Scophthalmus maximus L.) rhabdovirus (SMRV), termed PoHsp40A4, PoHsp40B6 and PoHsp40B11, whose encoded proteins all possess the conserved DnaJ domain, a signature motif of the Hsp40 family. Based on different protein structure and phylogenetic analysis, they can be categorized into two subfamilies, PoHsp40A4 for Type I Hsp40, PoHsp40B6 and PoHsp40B11 for Type II Hsp40. Further expression analysis revealed two very different types of kinetics in response either to heat shock or to virus infection, with a marked induction for PoHsp40A4 and a weak one for both PoHsp40B6 and PoHsp40B11. A very distinct tissue distribution of mRNA was also revealed among the three genes, even between PoHsp40B6 and PoHsp40B11. This is the first report on the transcriptional induction of Hsp40 in virally stimulated fish cells, and the differential expressions might reflect their different roles in unstressed and stressed cells. PMID- 16377210 TI - Molecular cloning and characterisation of peroxinectin, a cell adhesion molecule, from the giant freshwater prawn Macrobrachium rosenbergii. AB - Expression of peroxinectin cDNA was determined from haemocytes of giant freshwater prawn Macrobrachium rosenbergii using oligonucleotide primers and reverse transcription polymerase chain reaction (RT-PCR) based on the peroxinectin sequence of white shrimp Litopenaeus vannamei, tiger shrimp Penaeus monodon, and freshwater crayfish Pacifastacus leniusculus. The peroxinectin of M. rosenbergii was constitutively expressed. Analysis of the nucleotide sequence revealed that the cDNA clone has an open reading frame of 2,403 bp encoding a protein of 801 amino acids including a 20 amino acid signal peptide. The calculated molecular mass of the mature protein (781 amino acids) was 88.7 kDa with an estimated pI of 6.8. A putative peroxidase domain and a putative integrin binding motif, KGD (Lys-Gly-Asp) were observed in prawn peroxinectin at the C terminal. Sequence comparison showed that peroxinectin deduced amino acid of M. rosenbergii had an overall similarity of 62%, 64%, and 66% to that of P. leniusculus, P. monodon, and L. vannamei, respectively. Quantitative real-time PCR analysis showed that peroxinectin transcript in haemocyte of M. rosenbergii decreased significantly after 3, 6 and 12h injection with Lactococcus garvieae. PMID- 16377211 TI - Noradrenaline modulates the immunity of white shrimp Litopenaeus vannamei. AB - The total haemocyte count (THC), phenoloxidase activity, respiratory burst, superoxide dismutase (SOD) activity, phagocytic activity and clearance efficiency in response to pathogen Vibrio alginolyticus were measured when the white shrimp Litopenaeus vannamei (18.4 +/- 1.2 g) were injected individually with noradrenaline at 10(-8), 10(-7) and 10(-6) mol shrimp(-1). For the shrimp that received noradrenaline at 10(-8), 10(-7) and 10(-6) mol shrimp(-1), the THC decreased by 15%, 21% and 32%, phenoloxidase activity decreased by 15%, 31% and 31%, respiratory burst decreased by 13%, 21% and 32%, and SOD activity decreased by 46%, 56% and 55%, respectively, after 2 h. The phagocytic activity and clearance efficiency of shrimp that received noradrenaline at either dose decreased significantly after 2 h. The THC, phenoloxidase activity, respiratory burst, SOD activity, phagocytic activity and clearance efficiency returned to normal values after 4, 4, 8, 24, 16 and 8 h, respectively, in the shrimp that received noradrenaline at either dose. In another experiment, L. vannamei which had received noradrenaline at 10(-8), 10(-7) and 10(-6) mol shrimp(-1) were challenged after 1h by injection with V. alginolyticus at 1.0 x 10(5) colony forming units (cfu)shrimp(-1) and then placed in seawater of 20 per thousand. The cumulative mortality of shrimp that received noradrenaline at either dose was significantly higher than that of shrimp that received saline after 4 h, and at the termination of the experiment (48 h after the challenge). It is therefore concluded that noradrenaline administration at 10(-6) mol shrimp(-1) or less causes immune modulation of L. vannamei. PMID- 16377212 TI - Dynamics of indirect symptoms of skeletal muscle damage after stretch-shortening exercise. AB - Healthy untrained men (age 20.4+/-1.7 years, n=20) volunteered to participate in an experiment in order to establish dynamics of indirect symptoms of skeletal muscle damage (ISMD) (decrease in maximal isometric voluntary contraction torque (MVCT) and torque evoked by electrostimulation at different frequencies and at different quadriceps muscle length, height (H) of drop jump (DJ), muscle soreness and creatine kinase (CK) activity in the blood) after 100 DJs from 0.75 m height performed with maximal intensity with an interval of 20s between the jumps (stretch-shortening exercise, SSE). All ISMDs remained even 72 h after SSE (P<0.01-0.001). The muscle experienced greater decrease (P<0.01) in torque evoked by electrostimulation (at low stimulation frequencies and at short muscle length in particular) after SSE than neuromuscular performance (MVCT and H of DJ) which demonstrated secondary decrease (P<0.01) in neuromuscular performance during the first 48 h after SSE. Within 24-72 h after the SSE the subjects felt an acute muscle pain (5-7 points approximately) and the CK activity in the blood was significantly increased up to 1200 IU/L (P<0.001). A significant correlation between decrease in MVCT and H of DJ 24-48 h after SSE on the one hand and muscle soreness registered within 24-48 h after SSE on the other was observed, whereas correlation between the other indirect symptoms of skeletal muscle damage was not significant. PMID- 16377213 TI - Reflexes in prior polio and their relation to weakness and anterior horn cell loss. AB - The aim of this study was to evaluate the reflex pattern in patients with prior polio and to relate these findings to the degree of anterior horn cell (AHC) involvement and loss of muscle force. Twenty-five prior polio subjects were investigated with electromyography (EMG), force testing and reflex studies, which included the patellar and Achilles reflex, H-reflex, T-response and interlimb reflex (ILR). The clinical reflexes, H/M-ratio and T-response amplitude at rest were positively correlated with force and negatively correlated with the degree of AHC loss. The H/M-ratio was decreased compared with age matched controls. ILR was present in 68% of the prior polio patients but did not exist in controls. The presence of the ILR was not correlated with the degree of AHC loss or force. The reflex studies gave two main findings. The first is reduced excitability of monosynaptic connections in the motor neuron pool, which is related to weakness. The other is the presence of ILR as an indicator of interneuronal hyper excitability, which is not related to weakness. PMID- 16377214 TI - Gastrocnemius and soleus muscle length, velocity, and EMG responses to changes in pedalling cadence. AB - Several authors have shown different excitation patterns for soleus and gastrocnemius muscles in response to cadence manipulation during cycling. The purpose of this study was to examine gastrocnemius and soleus length and velocity change as a function of pedalling cadence to consider mechanisms underlying these excitation differences. Ten male and two female cyclists rode at five randomly assigned cadences (50, 65, 80, 95, and 110 rpm) at a nominal 200 W power output while EMG of the gastrocnemius and soleus and sagittal plane video were recorded. Joint-coordinate data for the knee and ankle were used with equations of Grieve et al. [Grieve D, Pheasant S, Cavanagh PR. Prediction of gastrocnemius length from knee and ankle joint posture, in: E. Asmussen, K. Jorgensen, editors. International Series on Biomechanics, vol. 2A, Baltimore: University Park Press; 1978. p. 405-412] to compute gastrocnemius and soleus length and velocity. Consistent with previous publications, gastrocnemius displayed a significant (p<0.05) increase in integrated EMG with increased cadence, whereas cadence had no significant effect on integrated EMG of the soleus. The ankle became significantly (p<0.05) more plantar flexed and reflected a reduced range of motion with increased cadence while the knee became significantly (p<0.05) less extended. Soleus decreased its range of motion by 29%, whereas gastrocnemius decreased its range of motion by 9%. In contrast, soleus increased its velocity range by 32% and gastrocnemius increased by 45%. These data show that with increased cadence gastrocnemius operated over a narrower range of operating lengths but at a higher range of shortening velocity than soleus. The higher range of velocity may have resulted in the need for a relatively higher excitation, as indicated by the integrated EMG, as the muscle was working at a different range on its force-velocity curve. During the recovery portion of the pedalling cycle, the soleus was acting eccentrically while the gastrocnemius acted concentrically indicating the triceps surae complex did not always act in unison. PMID- 16377215 TI - Asporin and knee osteoarthritis in patients of Greek origin. AB - Ostearthritis (OA) is characterized by focal areas of loss of the articular cartilage in synovial joints, associated with varying degrees of osteophyte formation, subchondral bone change and synovitis. The Asporin (ASPN) gene which encodes a protein of the extracellular cartilage matrix contains a triplet repeat encoding for aspartic acid (D) within exon 2 The D14 allele was found associated with knee and hip osteoarthritis in case-control study in the Japanese population. Genotyping Greek knee OA patients for the D repeats we determined that the D15 allele could be considered a risk allele for our population. PMID- 16377216 TI - The behavior of SATB1, a MAR-binding protein, in response to apoptosis stimulation. AB - As a MAR-binding protein, SATB1 regulates genes by folding chromatin into a loop domain. Apoptosis is known to be accompanied by a collapse of nuclear architecture and cleavage of condensing chromatin into oligonucleosomal fragments. To further understand the functional role of MAR-binding proteins during apoptosis we investigated the relationship of the behavior of SATB1 and the collapse of nuclear architecture in Jurkat cells with immunostaining and Western blot analysis. We demonstrated that SATB1 formed special three dimensional network distributions during early apoptosis. The distribution change of SATB1 was associated with cleavage of the protein and accompanied by the nuclear architecture collapse. Cleavage of SATB1 was mediated by caspase-3 and was apoptosis specific. Our observations further support the notion that early proteolysis of MAR-binding proteins might represent a universal mechanism that renders these DNA sites vulnerable to endonucleolysis. PMID- 16377217 TI - Nightmares: from anxiety symptom to sleep disorder. AB - The DSM-IV-TR definition of nightmares-extremely frightening dreams from which the person wakes up directly-is unnecessarily narrow. Other emotions (anger, grief) have also been reported in nightmares, and direct awakening from a bad dream seems to be unrelated to increased distress. In addition, assessment of nightmares is problematic. Polysomnographic recordings have an ameliorating effect on nightmare frequency, retrospective measurements tend to underestimate nightmare frequency, and persons with frequent nightmares may feel reluctant to fill out (daily) prospective measurements. For studying nightmares, it is necessary to distinguish idiopathic nightmares from posttraumatic nightmares, which are part of a posttraumatic stress reaction or disorder that may result from experiencing a traumatic event. Both types of nightmares have been associated with an elevated level of periodic limb movements, although only posttraumatic nightmares seem to be related to more and longer nocturnal awakenings. Nightmares have also been repeatedly associated with the general level of psychopathology, or the so-called personality factor neuroticism. Nightmare distress, the impact on daily functioning caused by nightmares, may function as a mediating variable. Several studies in the last decades have shown that nightmares can be treated with several cognitive-behavioral techniques. The cognitive-restructuring technique imagery rehearsal therapy is the treatment of choice for nightmares, although a randomized controlled trial with an attention control-group has not yet been carried out. Nightmares are more than a symptom of a larger (anxiety) syndrome and need to be viewed from a sleep medicine perspective: nightmares are a highly prevalent and separate sleep disorder that can and should receive specific treatment. PMID- 16377218 TI - Cryptic promoter activity in the coding region of the HMG-CoA reductase gene in Fusarium graminearum. AB - Head blight or scab disease caused by Fusarium graminearum poses a major threat to wheat and barley production in North America and other countries. To better understand the molecular mechanisms of F. graminearum pathogenesis, we have generated a collection of random insertional mutants. In mutant 222, one of the transformants significantly reduced in virulence, the transforming vector was inserted at amino acid 269 of the hydroxymethyl-glutaryl CoA reductase gene (HMR1) that encodes a key enzyme in sterol and isoprenoid biosynthesis. The N terminal transmembrane domains of HMR1 were disrupted, but the C-terminal catalytic domain was intact in mutant 222. We failed to isolate mutants deleted of the HMR1 gene, suggesting that HMR1 is an essential gene. Mutants deleted of the N-terminal 254 amino acids of HMR1 were viable and phenotypically similar to mutant 222. In both mutant 222 and the hmr1Delta254 mutants, a 3-kb truncated HMR1 transcript was detectable by northern blot analyses. In the wild-type strain, only the 5-kb messenger was observed. The initiation site of truncated HMR1 transcripts was determined by 5'-RACE to be 507bp upstream from the catalytic subunit. When a HMR1 fragment corresponding to the DNA sequence of HMR1269-641 was translationally fused to a promoter-less GFP construct, green fluorescent signals were detectable in vegetative hyphae of the resulting transformants. These data indicate that this region of HMR1 ORF has cryptic promoter activity and can express the catalytic domain in hmr1 mutants deleted of its N-terminal portion. Our results also illustrate the importance of the HMR1 gene and the function of its transmembrane domains in F. graminearum. PMID- 16377219 TI - Pharmacokinetics and milk penetration of moxifloxacin after intramuscular administration to lactating goats. AB - The pharmacokinetics of moxifloxacin was studied following intramuscular administration of 5mg/kg to healthy lactating goats (n=6). Moxifloxacin concentrations were determined by high performance liquid chromatography assay with fluorescence detection. The moxifloxacin plasma concentration versus time data could best be described by a one-compartment model. The plasma moxifloxacin clearance (Cl) was mean standard deviation (+/-SD) 0.49+/-0.14 L/h kg. The apparent volume of distribution (V(z)) was 0.83+/-0.20 L/kg. The terminal half life (t(1/2 lambda z)) was 1.31+/-0.64 h. Moxifloxacin penetration from blood to milk was rapid and the high AUC(milk)/AUC(plasma) and C(max-milk)/C(max-plasma) ratios reached indicated a good penetration of moxifloxacin into the milk. PMID- 16377220 TI - Plasma leptin, ghrelin and adiponectin concentrations in young fit racehorses versus mature unfit standardbreds. AB - Concentrations of hormones related to energy homeostasis may differ between populations with varied body compositions, acting as signals to increase or decrease energy intake and/or expenditure. How these parameters correlate with body composition in horses and how they vary in fit (F) versus unfit (UF) Standardbred racehorses is unclear. The purpose of this study was to test the hypothesis that plasma concentrations of glucose (GLU), insulin (INS), cortisol (CORT), ghrelin (GHRL), adiponectin (ADIP) and leptin (LEP) would be correlated with body composition and differ in fit (F) versus unfit (UF) horses. Fasting plasma samples were taken from 12 unfit (11 +/- 2 years, 521 +/- 77 kg; mean +/- SD) and 34 fit (4 +/- 2 years, 475 +/- 83 kg) Standardbred horses. GHRL, LEP, ADIP, INS and CORT concentrations were measured using radioimmunoassay. GLU concentration was measured using colorometric kits. Body composition data included body weight, body condition score (BCS), and percent fat (%fat) calculated using rump fat thickness measured ultrasonically and the Westervelt equation. Data were analyzed using Pearson Product moment and Student's t tests. There were no differences (P>0.05) between F and UF horses for the plasma concentrations of CORT (69 +/- 14 versus 76 +/- 23 microg/dL), INS (7.2 +/- 3.5 versus 7.1 +/- 1.8 microIU/mL) or GLU (90 +/- 6 versus 86 +/- 7 mg/dL). Plasma GHRL and ADIP concentrations were greater (P<0.05) in F versus UF horses (54 +/- 27 versus 33 +/- 17 pg/mL and 1820 +/- 276 versus 1333 +/- 249 ng/mL, respectively), while plasma LEP was lower in F versus UF (1.0 +/- 0.6 versus 4.4 +/- 2.4 ng/mL, P<0.001). BCS and %fat were lower in F versus UF horses (4.8 +/- 0.3 versus 6.7 +/- 0.5 and 11.9 +/- 1.6 versus 15.4 +/- 2.5%, respectively), with no correlation between %fat and GHRL (-0.12, P>0.05), although there was a positive correlation between %fat and LEP (+0.72, P<0.05), and a negative correlation between %fat and ADIP (-0.40, P<0.05). The data show that in comparing fit and unfit horses, there are variations in body composition as well as concurrent and substantial differences in the concentrations of hormones, cytokines, and other parameters related to the control of appetite and feed intake. PMID- 16377221 TI - Alleviating the pain caused by the castration of cattle. PMID- 16377222 TI - Involvement of CRF on the anorexic effect of GLP-1 in layer chicks. AB - Glucagon-like peptide-1 (GLP-1) is recognized as an anorexic peptide in the brain of chicks. However, the mechanism underlying the inhibition of feeding has not been well studied. It is reported that GLP-1 activates neurons containing corticotrophin-releasing factor (CRF) in the brain of mammals. Since CRF is also an anorexic peptide, it is possible that the anorexic effect of GLP-1 is mediated by CRF in chicks. The present study was carried out to test this. First, we determined plasma corticosterone (CORT) concentrations after intracerebroventricular (ICV) injection of GLP-1 and found that this treatment increased CORT release in layer chicks. The CORT-releasing effect was partly attenuated by co-injection of astressin, a CRF receptor antagonist, demonstrating that GLP-1 stimulated CORT secretion by activation of CRF neurons. CRF neurons also appear to be involved in mediating the inhibition of food intake by GLP-1 because this effect was also partly attenuated by astressin. Furthermore, we demonstrated that the anorexic effect of GLP-1 was weaker in broiler than layer chicks. The present results suggest that the anorexic effect of GLP-1 might be mediated by CRF neurons in the chick brain and that the sensitivity of the inhibitory response to GLP-1 differs between chick strains. PMID- 16377223 TI - Energy, quiescence and the cellular basis of animal life spans. AB - Animals are routinely faced with harsh environmental conditions in which insufficient energy is available to grow and reproduce. Many animals adapt to this challenge by entering a dormant, or quiescent state. In some animals, such as the nematode Caenorhabditis elegans, quiescence is coincident with increased stress resistance and longevity. Here we review evidence that the rules of life span extension established in C. elegans may be generally true of most animals. That is, that the rate of animal aging correlates inversely with cellular resistance to physiological stress, particularly oxidative stress, and that stress resistance is co-regulated with the quiescence adaptation (where the latter occurs). We discuss evidence for highly conserved intracellular signalling pathways involved in energy sensing that are sensitive to aspects of mitochondrial energy transduction and can be modulated in response to energetic flux. We provide a broad overview of the current knowledge of the relationships between energy, metabolism and life span. PMID- 16377224 TI - Serotonin: a coordinator of feeding-related physiological events in the blood gorging bug, Rhodnius prolixus. AB - Rhodnius prolixus is an obligatory blood-feeder that can ingest blood meals of up to 10 times its mass. Rapid production of urine commences within 2-3 min of the start of feeding in order to eliminate the load of water and salts, and so there is an increase of Malpighian tubule secretion greater than 1,000 fold in response to feeding. Feeding and post-prandial diuresis in Rhodnius are highly coordinated events, including for example, host recognition, probing, injection of saliva, cuticle plasticization, passage of blood through the digestive system, diuresis and excretion. This review illustrates that many of the known functions of serotonin in Rhodnius are feeding-related. Serotonin coordinates or 'orchestrates' feeding-related physiological events either as a neurotransmitter/neuromodulator, delivered to target tissues in the nerve supply, or as a neurohormone, delivered by the haemolymph. Thus, serotonin has physiological effects upon the salivary glands, cuticle, digestive tract, cardiac muscle, and Malpighian tubules. By discussing these aspects, the review illustrates that serotonin acts in a coordinated manner to prepare Rhodnius for this energy-demanding process of feeding and diuresis. PMID- 16377225 TI - Effects of diet on the carotenoid pigment and lipid content of Pomacea bridgesii as determined by quantitative high performance thin layer chromatography. AB - High performance thin layer chromatography was used to quantify the concentrations of beta-carotene and lutein in the whole bodies and digestive gland-gonad complexes of Pomacea bridgesii fed on a lettuce diet and a hen's egg yolk diet, and snails starved for one week. beta-carotene and lutein concentrations were determined in the fecal matter of cultures on the lettuce and yolk diets as well. Significantly higher amounts of beta-carotene were found in the whole bodies of snails fed lettuce compared with those starved for a week. beta-carotene was not detected in the whole bodies and digestive gland-gonad complexes of snails fed hen's egg yolk. Neutral and polar lipid levels were also quantified in P. bridgesii fed lettuce and yolk. Significantly higher amounts of triacylglycerols were detected in the digestive gland-gonad complexes of snails maintained on the egg yolk diet. PMID- 16377226 TI - Riboflavin-responsive glutaryl CoA dehydrogenase deficiency. AB - We report here riboflavin responsiveness in a patient with glutaryl CoA dehydrogenase (GCDH) deficiency, compound heterozygous for the S139L and P248L mutations and with 20% residual GCDH enzyme activity in vitro. Our results suggest the mitochondrial GCDH homotetramer remains intact with one of these mutations associated with the binding site of the single FAD cofactor and that pharmacological doses of the cofactor precursor may be sufficient to induce an increase in activity in the mutant GCDH enzyme, although not sufficient to normalise urinary organic acid excretion. Serine139 is one of nine conserved amino acid residues that line the binding site of the protein and is in close proximity to both substrate and FAD cofactor. It is possible that steric alterations caused by substitution of serine with leucine at this position may be overcome with high cofactor concentrations. P248L is also associated with some residual GCDH activity in other patients and the unique combination of S139L with P248L may also explain the results in our patient. Responsiveness to riboflavin in our patient has been compared with two other patients with glutaric aciduria type 1 and minimal residual GCDH activity, one with homozygosity for the R257Q mutation and one with heterozygosity for the G354S mutation and a novel G156V mutation. A low lysine diet reduced glutaric acid excretion in our riboflavin responsive GCDH-deficient patient almost to control values. She is now 21 years of age and clinically and neurologically normal. PMID- 16377227 TI - Modulation of 6-phosphofructo-1-kinase oligomeric equilibrium by calmodulin: formation of active dimers. AB - Muscle 6-phospho-1-kinase (PFK) is the key regulatory enzyme of the glycolytic pathway and is a calmodulin-binding protein binding two calmodulin molecules per PFK protomer. This enzyme is characterized by a complex regulation that involves its allosteric behavior modulated by several ligands, which modulate the equilibrium between the active tetramers and the inactive dimers of the enzyme. Calmodulin is described to induce the dimerization of PFK, so inhibiting its catalytic activity. Here, we show that binding of calmodulin specifically to its higher-affinity site of PFK induce its dimerization without compromising enzyme catalytic activity forming a hitherto not described active dimmer of PFK. It is also shown that the dimerization is a Ca2+ -dependent event that responds to physiological intracellular Ca2+ concentrations and decrease the interaction of the enzyme to membrane site, which stimulate its catalytic activity. We propose that the effects of calmodulin on PFK reported here are of great physiological significance due to the response to physiological concentrations of Ca2+ and due to be in accordance to the known effects of calmodulin on cell ATP production. We also propose that calmodulin might affect the interaction of PFK to other cellular components as the cytoskeleton. PMID- 16377228 TI - Vaccination strategies to prevent tuberculosis in the new millennium: from BCG to new vaccine candidates. AB - Current global control efforts targeting tuberculosis (TB) include the treatment of latent TB infection, case detection and treatment with directly observed therapy short-course (DOTS), and BCG (bacille Calmette-Guerin) vaccination. However, BCG has been found to decrease only childhood TB morbidity and mortality but has a very limited effect in the transmission dynamics of the infection. These limitations of BCG are the driving force for the development of new TB vaccines. New TB vaccine candidates have entered clinical evaluation and many more are in the pipeline to undergo clinical testing. New vaccine candidates may offer better protection than that afforded by currently available BCG vaccines. Furthermore, combined vaccination schedules against TB seem to be a promising strategy in the new millennium. PMID- 16377229 TI - Rheumatoid nodulosis. Two cases with destructive polyarthritis after 20 years. AB - The term "rheumatoid nodulosis" was coined by Ginsberg in 1975 to designate a rare and distinctive form of rheumatoid disease. Anecdotal case reports suggest a benign nondestructive course, although prolonged follow-up data are usually unavailable. We describe two cases of typical rheumatoid nodulosis with follow ups exceeding 25 years. Onset occurred at 14 and 22 years of age, respectively. Both patients presented with palindromic rheumatism, positive tests for rheumatoid factors, negative tests for other biological markers, and normal radiographs. Multiple subcutaneous nodules developed after 4 and 6 years with palindromic flares, respectively. Functional impairments and disfigurement required several surgical procedures to remove nodules. Histology was typical for rheumatoid nodules. Neither patient responded to disease-modifying anti-rheumatic drugs (gold, antimalarials, and D-penicillamine). Treatment consisted of nonsteroidal anti-inflammatory drugs combined with prednisone as needed. After 20 and 22 years of follow-up, respectively, both patients had typical rheumatoid arthritis with deformities and radiological joint destruction. In conclusion, these two cases establish that rheumatoid nodulosis can occur as a presentation of rheumatoid arthritis with a potential for severe joint damage after many years. PMID- 16377230 TI - Animal models of HLA-B27-associated diseases: new outcomes. AB - The HLA-B27 molecule is strongly associated with the spondyloarthropathies, a group of chronic inflammatory diseases, affecting the skeleton, the bowel and the skin. This association has been largely studied, but mechanisms of pathology remain unclear. The HLA-B27 transgenic rats develop a spontaneous disease that strikingly resembles human spondyloarthropathies, dependent of bacterial flora and implicating the immune system. The presence of CD4+ T cells is required, and antigen presenting cells (APC) expressing high levels of HLA-B27 likely play an important role. Indeed, APC are defective in naive T lymphocytes stimulation. This default appears to implicate the APC/T cells contact, and may result in a loss of tolerance toward microbial flora. Two models of skeletal inflammation linked to HLA-B27 have been developed in mice. The ANKENT mice develop a spontaneous ossifying enthesitis affecting ankle and tarsal joints, with increased frequency in the presence of an HLA-B27 transgene. The HLA-B27 transgenic mice lacking endogenous beta2 microglobulin develop arthritis of hind paws. In this model, homodimers of B27 heavy chains could be implicated in the pathogenesis by presenting exogenous peptides to CD4+ T cells. PMID- 16377231 TI - Comparison of enhancement of pentachlorophenol sonolysis at 20 kHz by dual frequency sonication. AB - The comparison of enhancement effect of pentachlorophenol sonolysis at 20 kHz by different dual-frequency ultrasonic irradiations has been investigated. Dual frequency (20 kHz/40 kHz, 20 kHz/530 kHz, 20 kHz/800 kHz and 20 kHz/1040 kHz) ultrasounds have been used. It has been found that the rate of pentachlorophenol degradation at dual-frequency ultrasonic irradiation is the highest compared to mono-frequency ultrasonic systems. The combination of dual-frequency systems has synergistic effect and the enhancement effect of sonochemical degradation of pentachlorophenol at 20 kHz by dual-frequency systems appears to be remarkable frequency sensitive. The order of contribution to the enhancement effect of sonochemical degradation of pentachlorophenol at 20 kHz is as follows: 530 kHz > 800 kHz > 40 kHz > 1040 kHz. PMID- 16377232 TI - The role of cytokines and trophic factors in epiretinal membranes: involvement of signal transduction in glial cells. AB - Idiopathic epiretinal membranes (ERMs) in the macular region can cause a reduction in vision and sometimes recurs after surgical removal, but its pathogenic mechanisms are still unknown. On the other hand, the presence of secondary ERMs has been associated with various clinical conditions including proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR). Recent studies have shown a significant association between clinical grades of PDR or PVR, and the expression levels of specific cytokines and/or growth factors in the vitreous fluid. Expression of these factors and their receptors are also observed in secondary ERMs. ERMs are composed of many cell types such as retinal pigment epithelial cells and vascular endothelial cells, however the role of glial cells is yet unclear. Interestingly, glial cells in ERMs express some trophic factor receptors and transcription factors, such as NF kappaB, suggesting an involvement of glial signal transduction in the pathogenesis of ERMs. In this review, we summarize recent progress regarding the clinical and laboratory findings of ERMs. PMID- 16377233 TI - Targeted tumor therapy with the TGF-beta 2 antisense compound AP 12009. AB - TGF-beta overexpression is a hallmark of various malignant tumors. This is due to the pivotal role of TGF-beta as it regulates key mechanisms of tumor development, namely immunosuppression, metastasis, angiogenesis, and proliferation. We have developed a new immunotherapeutic approach for the treatment of malignant tumors based on the specific inhibition of TGF-beta2 by the antisense oligodeoxynucleotide AP 12009. After providing preclinical proof of concept, we assessed safety and efficacy of AP 12009 in clinical phase I/II open-label dose escalation studies in high-grade glioma patients. Median survival time after recurrence exceeded the up to date literature data for chemotherapy. A phase I/II study in pancreatic carcinoma and malignant melanoma is currently ongoing. Our results implicate targeted TGF-beta2 suppression as a promising therapeutic approach for malignant tumor therapy. PMID- 16377234 TI - Spectroscopic and conductometric studies of molecular complex formation between 2,4,6-trinitrophenol and diaza-18-crown-6, tetraaza-14-crown-4 and cryptand C222 in 1,2-dichloroethane solution. AB - The formation of molecular complexes between 2,4,6-trinitrophenol (TNP) and aza substituted macrocycles diaza-18-crown-6 (DA18C6), tetraaza-14-crown-4 (TA14C4) and cryptand C222 in 1,2-dichloroethane solution was investigated spectrophotometrically and conductometrically. The mole ratio and continuous variations studies based on both physicochemical techniques employed clearly revealed the formation of both 1:1 and 2:1 (TNP:macrocycle) adducts in solution. Formation of the resulting complexes was also confirmed by 1H NMR and IR spectroscopic studies. Formation constants of the resulting complexes were evaluated from computer fitting of the corresponding mole ratio data. Stability of the resulting complexes was found to vary in the order C222 approximately TA14C4 > DA18C6. PMID- 16377235 TI - Health search engine with e-document analysis for reliable search results. AB - OBJECTIVE: After a review of the existing practical solution available to the citizen to retrieve eHealth document, the paper describes an original specialized search engine WRAPIN. METHOD: WRAPIN uses advanced cross lingual information retrieval technologies to check information quality by synthesizing medical concepts, conclusions and references contained in the health literature, to identify accurate, relevant sources. Thanks to MeSH terminology [1] (Medical Subject Headings from the U.S. National Library of Medicine) and advanced approaches such as conclusion extraction from structured document, reformulation of the query, WRAPIN offers to the user a privileged access to navigate through multilingual documents without language or medical prerequisites. RESULTS: The results of an evaluation conducted on the WRAPIN prototype show that results of the WRAPIN search engine are perceived as informative 65% (59% for a general purpose search engine), reliable and trustworthy 72% (41% for the other engine) by users. But it leaves room for improvement such as the increase of database coverage, the explanation of the original functionalities and an audience adaptability. CONCLUSION: Thanks to evaluation outcomes, WRAPIN is now in exploitation on the HON web site (http://www.healthonnet.org), free of charge. Intended to the citizen it is a good alternative to general-purpose search engines when the user looks up trustworthy health and medical information or wants to check automatically a doubtful content of a Web page. PMID- 16377236 TI - A proposal of the modified liver damage classification for hepatocellular carcinoma. AB - OBJECTIVE: The purpose of this study was to evaluate the validity of the two common classifications for assessing liver function; the Child-Pugh classification and the liver damage classification. We also examined the feasibility of the modified liver damage classification. METHODS: A total of 2306 HCC patients diagnosed between 1990 and 2002 were categorized according to the three classifications. The modified liver damage classification is calculated by summation of the scores for five variables (serum bilirubin level, serum albumin level, prothrombin activity, ICG retention rate at 15min, ascites) of the liver damage classification and classified patients into grades A-C in the similar manner as the Child-Pugh classification. The differences in distribution and survival rate of the patients in each group were compared. RESULTS: With respect to the patient distribution, 1787 (77.5%) and 469 (20.3%) patients were categorized into Child-Pugh grades A and B, respectively, whereas 1187 (51.5%) and 962 (41.7%) patients were categorized into liver damage grades A and B, respectively. As a result of this disproportionate distribution, survival rates of Child-Pugh grades A and B were lower than those of the liver damage grades A and B. Furthermore, some discrepancies were found in the distribution of patients between the liver damage classification and the modified liver damage classification. One hundred and forty-one patients of the 1187 liver damage grade A patients were categorized into grade B by the modified liver damage classification and 71 patients of the 962 liver damage grade B patients were categorized into grade C by the modified liver damage classification. The survival rates of these subgroups were similar to those of the liver damage grades B and C, respectively. CONCLUSION: The modified liver damage classification appears to be the best available tool for assessing residual liver function, for estimating the survival of Japanese HCC patients and for making decisions concerning the treatment of these patients. PMID- 16377237 TI - Th1 response during ribavirin and interferon-alpha combination therapy in chronic hepatitis C. AB - Ribavirin and interferon-alpha induce Th1 polarization of human CD4+ T cells. The study was conducted to investigate the whether cellular immune response during ribavirin/interferon-alpha therapy is associated with viral eradication by examining mRNA expression of molecules relevant to Th1 and Th2 polarization in CD4+ cells of 13 patients with chronic hepatitis C (seven patients with sustained viral response and six with transient response). Peripheral CD4+ T lymphocytes at 0, 4 and 24 weeks of treatment were tested. There were no significant differences in the mRNA levels at each point of time of the treatment between patients with sustained viral response and those with transient response. The percent increase in mRNA level of the IL-12R beta2 chain from the baseline to the end of the treatment was significantly higher in patients with sustained viral response (15.3+/-6.1%) than in those with transient response (-1.6+/-4.7%, p<0.05). There was no significant difference in percent changes in level of IL-12R beta1 chain mRNA between the two groups. In conclusion, the results of this study indicate that the increase of Th1 response is related to the inflammatory activity in the liver and possibly to ribavirin and interferon-alpha therapy. It is also suggested that the measurement of Th1 response has the potential to distinguish patients with relapse from those with sustained virus response. PMID- 16377238 TI - Sources of cortical rhythms change as a function of cognitive impairment in pathological aging: a multicenter study. AB - OBJECTIVE: The present study tested the hypothesis that cortical electroencephalographic (EEG) rhythms. change across normal elderly (Nold), mild cognitive impairment (MCI), and Alzheimer's disease (AD) subjects as a function of the global cognitive level. METHODS: Resting eyes-closed EEG data were recorded in 155 MCI, 193 mild AD, and 126 age-matched Nold subjects. EEG rhythms of interest were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5-13 Hz), beta 1 (13-20 Hz), and beta 2 (20-30 Hz). EEG cortical sources were estimated by LORETA. RESULTS: Occipital delta and alpha 1 sources in parietal, occipital, temporal, and 'limbic' areas had an intermediate magnitude in MCI subjects compared to mild AD and Nold subjects. These five EEG sources presented both linear and nonlinear (linear, exponential, logarithmic, and power) correlations with the global cognitive level (as revealed by mini mental state examination score) across all subjects. CONCLUSIONS: Cortical EEG rhythms change in pathological aging as a function of the global cognitive level. SIGNIFICANCE: The present functional data on large populations support the 'transitional hypothesis' of a shadow zone across normality, pre-clinical stage of dementia (MCI), and AD. PMID- 16377239 TI - CRT improves the exercise capacity and functional reserve of the failing heart through enhancing the cardiac flow- and pressure-generating capacity. AB - BACKGROUND: While information on how cardiac resynchronisation therapy (CRT) affects cardiac performance at rest is readily available, the mechanisms whereby CRT alters cardiac function during maximal exercise are unclear. AIMS: We examined the medium-term effects of CRT on cardiac and physical functional reserve of patients with severe heart failure (CHF) and prolonged QRS duration. METHODS: Seventeen consecutive patients with severe CHF (NYHA III-IV) and widened QRS underwent maximal cardiopulmonary exercise testing with non-invasive central haemodynamic measurements before and 6-8 weeks after CRT pacemaker implantation. RESULTS: After CRT there were significant increases in exercise cardiac output by 19.3% (P=0.0048) from 9.5+/-3.4 l min(-1), peak mean arterial blood pressure by 14.1% (P=0.0001) from 91.3+/-13.6 mm Hg, and peak cardiac power output by 37.2% (P=0.0008) from 1.92+/-0.74 W. There were no significant changes in these variables at rest. Exercise duration (+42.3%, P=0.0002), NYHA functional class (P=0.0001) and SF-36 symptom score (P=0.0006) were also significantly improved. Powerful surrogate indicators of prognosis were also significantly improved with CRT: peak O(2) consumption (+20.9%, P=0.0007), VE/VCO(2) slope (-20.0%, P=0.005) and circulatory power (+42.0%, P=0.0012). CONCLUSION: In this cohort of patients, post-implant CRT significantly improved the flow-, pressure- and power-generating capacity of the failing hearts. This may be causally related to the improvements observed in exercise capacity, functional class and symptom scores. PMID- 16377240 TI - The expression of the virulence-associated effector protein gene avrA is dependent on a Salmonella enterica-specific regulatory function. AB - Although the avrA gene is prevalent among 80% of the Salmonella enterica serovars, only a small number of them usually express the respective virulence associated effector protein AvrA. However, under culture conditions below pH 6.0 many of the AvrA non-producer strains (e.g. S. Agona, S. Bovismorbificans, S. Virchow) begin to produce AvrA, while some others remain silent. Four phenotypical classes of S. enterica were identified under defined standard culture conditions: class 1 comprises strains with a constitutive synthesis of AvrA; class 2 comprises strains with an acid induction of AvrA; class 3 comprises strains with silent avrA genes; and the fourth class comprises strains which do not contain the avrA gene (class 0 strains). The expression of avrA was found to be controlled by a Salmonella-specific mechanism because cloned avrA genes from classes 1, 2, and 3 strains remain silent in Escherichia coli strains, while easily expressed in S. enterica strains. The expression of AvrA in classes 1, 2, and 3 strains does not coincide with the nucleotide sequences of the respective promoter or structural regions of the avrA genes, but depends directly on this Salmonella-specific regulatory system which appears to be differently modulated in the distinct Salmonella serovars. PMID- 16377241 TI - Constipation in paediatric oncology. AB - Constipation is a common problem encountered by many children during treatment for childhood cancer. It can be distressing and impact on the quality of life for the child. However, the advice and information they and their families receive can vary enormously. The clinical practice group (a subgroup of the paediatric oncology nurses forum (PONF)) decided to examine the management of constipation throughout the paediatric oncology units in the UK. This paper presents the findings and the subsequent action taken and highlights the need for further work. PMID- 16377242 TI - Glutamate- and GABA-based CNS therapeutics. AB - Glutamate- and GABA-releasing neurons form the basis for neurotransmission in the mammalian central nervous system (CNS). The co-ordination of these excitatory and inhibitory systems, together with intrinsic voltage-gated ion channels and G protein-coupled receptor modulation, provides the diverse neuronal firing patterns, network activity and synaptic plasticity that are required for the complexity of CNS function. Virtually all of the known molecular components of the gamma-aminobutyric acid (GABA) and glutamate neurotransmitter systems have been considered as potential therapeutic targets. Positive allosteric modulators of GABAA receptors, such as the benzodiazepines, have found wide clinical use, and the N-methyl-D-aspartate receptor antagonists ketamine and memantine have therapeutic utility. In these fundamental neurotransmitter systems, drugs that provide allosteric modulation of ligand-gated ion channels or G-protein-coupled receptors, or seek to selectively target receptor subtypes, appear to hold the greatest promise for the desired balance of efficacy and tolerability. This might also be achieved through targeting transporter subtypes. A large number of compounds based on these strategies are currently in clinical trials for diseases that span a wide range of CNS disorders. PMID- 16377244 TI - Serum lipoproteins: Trojan horses of the immune response? AB - T cells recognizing lipid antigens presented by CD1 molecules have an important role in the immune response. Several lipid antigens for CD1-restricted T cells have been identified, as have some rules of CD1 loading and CD1-restricted presentation. Little is known, however, about the delivery of lipid antigens from either extracellular compartments or CD1-negative cells to CD1-expressing antigen presenting cells (APCs). A recent study provides evidence for a role for apolipoprotein E in binding lipid antigens and delivering them to APCs. PMID- 16377245 TI - Mitochondrial genomes of parasitic nematodes--progress and perspectives. AB - Mitochondria are subcellular organelles in which oxidative phosphorylation and other important biochemical functions take place within the cell. Within these organelles is a mitochondrial (mt) genome, which is distinct from, but cooperates with, the nuclear genome of the cell. Studying mt genomes has implications for various fundamental areas, including mt biochemistry, physiology and molecular biology. Importantly, the mt genome is a rich source of markers for population genetic and systematic studies. To date, more than 696 mt genomes have been sequenced for a range of metazoan organisms. However, few of these are from parasitic nematodes, despite their socioeconomic importance and the need for fundamental investigations into areas such as nematode genetics, systematics and ecology. In this article, we review knowledge and recent progress in mt genomics of parasitic nematodes, summarize applications of mt gene markers to the study of population genetics, systematics, epidemiology and evolution of key nematodes, and highlight some prospects and opportunities for future research. PMID- 16377246 TI - Doubts about Trypanosoma equiperdum strains classed as Trypanosoma brucei or Trypanosoma evansi. PMID- 16377247 TI - The re-emergence of trichinellosis in China? PMID- 16377248 TI - The pitfalls of assigning disease syndromes to pathogens and vice versa. PMID- 16377249 TI - Entomopathogenic fungi as biological insecticides to control malaria. AB - Malaria is arguably the most serious vector-borne disease worldwide. The already alarming number of deaths caused by malaria is increasing, caused in part by the increase in mosquito resistance to chemical insecticides. In two recent articles, the use of an approach was reported that could open a new front in the fight against malaria. Laboratory and field studies demonstrate that entomopathogenic fungi can efficiently kill adult anopheline mosquitoes, the females of which are the obligatory vectors for malaria parasites. PMID- 16377250 TI - Red blood cell age determines the impact of storage and leukocyte burden on cell adhesion molecules, glycophorin A and the release of annexin V. AB - The influence of the age of the red blood cell (RBC) within its 120-day lifecycle at the time of blood donation on the RBC storage lesion is not well understood. Expression of cell adhesion molecules (CAMs) (CD44, CD47, CD58 and CD147), glycophorin A (GPA) and phosphatidylserine (PS) on young and old RBCs density separated prior to storage of the RBC concentrate was determined by flow cytometry. Older RBCs showed significantly reduced expression of GPA throughout storage and CD44 and CD147 from Day 28 onwards compared to young RBCs. Storage in the presence of leukocytes caused a significant decline in the expression of CD44, CD58, CD147 and GPA, whereas RBCs that were pre-storage leukocyte depleted maintained a relatively consistent level of expression throughout storage. PS was not detected at the external RBC membrane of young or old RBCs during storage. Increased levels of annexin V were detected in the supernatant of RBCs stored in the presence of leukocytes, with significantly greater supernatant levels found for old RBCs compared to young RBCs. These findings provide new insight into the RBC storage lesion and indicate that RBC age at the time of donation impacts upon the quality of stored RBC concentrates. PMID- 16377251 TI - The health status and quality of life of adults with X-linked agammaglobulinemia. AB - Forty-one adults (mean age 33) with a definitive diagnosis of X-linked agammaglobulinemia (XLA) completed a questionnaire concerning current and past medical problems and quality of life. Thirty-six of the 41 were working full time or were full time students; 18 had not missed any work or school due to infection in the previous year. Their quality of life was equivalent to that of the general US male population. Thirteen of the 41 reported that they had chronic lung disease, and 33 indicated that they had one or more episodes of sinusitis in the preceding year. Arthritis, diarrhea and skin infections were common but not debilitating. The 41 study subjects were more likely to have a prior family history of XLA, and they were more likely to have milder mutations in Btk, the gene responsible for XLA. These results indicate that most adults with XLA are moderately healthy and lead productive lives. PMID- 16377252 TI - Innate cellular immune responses in newborns. AB - Innate immunity assures the first line of defense against pathogenic microorganisms. Innate immune responses induced by bacteria, fungi, or viral replication are triggered by granulocytes, monocytes, macrophages, dentritic cells, and natural killer cells. Neonatal deficiency of innate cellular immunity includes a decreased production of interferons, IL-12/IL-23, and IL-18, and other proinflammatory cytokines, an impaired type-1 response of macrophages to IFN gamma, the most potent macrophage-activating agent in vivo, and to lipopolysaccharide, the primary constituent of the outer membrane of Gram negative bacteria. An increasing body of evidence suggests impaired responses of neonatal monocytes and macrophages to multiple TLR ligands. This review will discuss recent advances in understanding innate cellular immunity in human neonates, with respect to selected aspects of immune functions that may be related to increased susceptibility to infections. Components of TLR signaling and the immune consequence that may result from neonatal deficiencies will be highlighted. A better understanding of innate immunity can make the development of techniques possible by which physicians more accurately tailor prevention and treatment of neonatal infections. PMID- 16377253 TI - Subjective preference for lamotrigine or topiramate in healthy volunteers: relationship to cognitive and behavioral functioning. AB - OBJECTIVE: Outcomes research emphasizes patient self-assessment and preferences in optimizing treatment. We previously showed that lamotrigine produces significantly less cognitive and behavioral impairment compared with topiramate. In the current study we extend these observations to subject self-report of preference for lamotrigine or topiramate independent of potentially confounding effects of seizures or seizure control. Additionally, drug preference was related to effects of lamotrigine and topiramate on objective neuropsychological tests as well as self-perception on behavioral instruments. METHODS: Thirty-seven healthy volunteers completed a double-blind, randomized crossover design incorporating two 12-week treatment periods of lamotrigine and topiramate each titrated to a dose of 300 mg/day. Evaluation of 23 objective neuropsychological and 15 subjective behavioral measures occurred at four times: pretreatment baseline, first treatment, second treatment, and posttreatment baseline. Preference for lamotrigine or topiramate was assessed, while blinding was maintained, at the final study visit when each subject was asked which drug he or she would prefer to take. RESULTS: A large majority (70%) preferred lamotrigine, 16% stated preference for topiramate, and 14% had no preference (drugs equivalent). Consistent with preference, those preferring lamotrigine performed better on 19 of 23 objective and 13 of 15 subjective behavioral measurements while on lamotrigine. Inconsistent with preference, subjects preferring topiramate performed better on 19 of 23 objective and 9 of 15 subjective behavioral measures while on lamotrigine. Topiramate preference also did not correlate with IQ, serum concentration, body mass index, age, or gender. Topiramate preference did relate to responses on the Profile of Mood States. CONCLUSION: Lamotrigine was preferred by the majority of subjects, congruent with objective neuropsychological and subjective behavioral measures. In contrast, for those stating a preference for topiramate the results on objective neuropsychological measures were impaired while fewer complaints were noted on the Profile of Mood States. This suggests that preference for topiramate may be determined by an effect on mood. PMID- 16377254 TI - Evaluation of the lethal and sub-lethal toxicity and potential endocrine disrupting effect of nonylphenol on the zebra mussel (Dreissena polymorpha). AB - Nonylphenol (NP) is commonly found in surface waters nearby municipal wastewater treatment plants and was shown to have endocrine disrupting effects in aquatic organisms. The purpose of this study was to investigate the toxicity and potential endocrine disrupting effects of NP on the freshwater zebra mussel (Dreissena polymorpha). Toxicity assessment yielded LC(50) values of 3.68, 2.19 and 1.62 mg L(-1) after 15, 35 and 50 days of exposure, respectively. LC(10) values of 1.6, 1.11 and 0.68 mg L(-1) were respectively obtained for similar exposure periods. At concentrations >5 mg L(-1), mortality effects were significant, as were those relating to attachment and siphon extension (indicating filtration), both general indicators of health. Endocrine disruption effects were investigated after a prolonged exposure (112 d) to 5 and 500 microg L(-1) NP by measuring Vitellin (Vn)-like protein levels using the alkali-labile phosphate (ALP) assay and gel electrophoresis (GE). An increase in ALP levels was observed in both male and female mussels, although only marginal owing to a significant decrease in the mussels' health indicated by its condition, during the experiment. These levels, however, increased proportionally with NP concentration. Using solid phase thin-layer chromatography, we confirmed increased levels of the steroid cholesterol and evidence of NP uptake. Cholesterol levels in gonad tissue proved to be a more responsive biomarker of exposure to NP than levels of ALP. Further implications relating to the occurrence of endocrine disruption in the zebra mussel are discussed. PMID- 16377255 TI - Cadmium toxicity related to cysteine metabolism and glutathione levels in frog Rana ridibunda tissues. AB - The level of glutathione and sulfane sulfur and sulfurtransferases activity in adult frogs Rana ridibunda were investigated after the exposure to 40 mg or 80 mg CdCl(2) L(-1) for 96 h or 240 h. Cd accumulation in the liver, kidneys and testes was confirmed, and the highest Cd level was found in the testes. In the liver, the exposure to Cd resulted in an increase of GSH level and the activity of rhodanese, while the activity of 3-mercaptopyruvate sulfurtransferase and cystathionase decreased. The kidneys and brain showed the elevated level of GSH and the activity of all investigated sulfurtransferases, as well as sulfane sulfur especially in brain. In such tissues as the testes, muscles and heart, the level of GSH and the activity of 3-mercaptopyruvate sulfurtransferase were significantly diminished. The increased level of sulfane sulfur was determined in the testes and muscles and the increased activity of rhodanese in the testes and the heart. These findings suggest the possible role of sulfane sulfur and/or sulfurtransferases in the antioxidation processes, which can be generated in cells by cadmium. PMID- 16377256 TI - An evaluation of cobalt chloride as an O2-sensitive chemoreceptor stimulant in channel catfish. AB - The effects of cobalt chloride on heart rate, blood pressure, ventilatory frequency and opercular pressure amplitude in channel catfish, Ictalurus punctatus were measured to evaluate the potential of cobalt as a histochemical probe to study mechanisms of oxygen chemoreception, as well as assess the general effects of cobalt on the cardioventilatory physiology of fishes. Cobalt, like cyanide, has been previously used to stimulate oxygen chemoreceptors and hypoxic reflexes in mammals but there is little information on the cardioventilatory effects of cobalt on fish. Catfish were exposed to increasing concentrations (1 20 mg/kg) of cobalt in the water (external) or injections into the dorsal aorta (internal) and the cardioventilatory effects recorded. Mean arterial pressure showed a significant, dose-dependent increase in response to cobalt injections. Heart rate increased slowly, but significantly after cobalt injections but the magnitude of change was not dose-dependent. There was a small increase in ventilatory rate but no effect on amplitude. External cobalt had similar effects but the responses were weaker. Although cobalt stimulated some cardioventilatory reflexes the pattern and magnitude of the responses were noticeably different from those of cyanide and hypoxia. The results suggest that the cardioventilatory reflexes stimulated by cobalt were not mediated by O(2)-sensitive chemoreceptors and that cobalt is not an effective O(2) receptor stimulant in fishes. PMID- 16377257 TI - Spatio-temporal patterns of intestine-specific transcription factor expression during postnatal mouse gut development. AB - The small intestine matures from a primitive tube into morphologically and functionally distinct regions during gut development. Maximal expression of the genes encoding the digestive enzymes lactase-phlorizin hydrolase and sucrase isomaltase is spatially restricted to distinct segments along the anterior posterior axis of the small intestine and is temporally regulated during postnatal maturation. Transcription factors capable of interacting with the intestinal lactase and sucrase gene promoters are candidate regulators of spatio temporal patterning during gut development and maturation. We aimed to quantitatively examine and compare the relative expression levels of a set of intestine-specific transcription factors along the anterior-posterior gut axis during postnatal maturation. Our analysis was focused on the transcription factors capable of regulating the intestinal lactase and sucrase-isomaltase genes. A real-time PCR protocol was used to quantitatively examine and compare spatially and temporally the relative transcript abundance levels for intestine specific factors during postnatal intestinal maturation. Distinct spatial expressions patterns were detected along the length of the small intestine for PDX-1, Cdx-2, GATA-4, GATA-5, GATA-6, HNF-1alpha, HNF-1beta and CDP transcription factor genes. There is a general decline in transcript abundance for the factor genes during postnatal maturation. Defining the spatio-temporal expression patterns for intestine-specific transcription factor genes contributes to investigation of the roles that factor gradients play in mediating gut development and differentiation. PMID- 16377258 TI - CyNodal, the Japanese newt nodal-related gene, is expressed in the left side of the lateral plate mesoderm and diencephalon. AB - The nodal and nodal-related genes play fundamental roles during deuterostome left right axis formation. Several of these genes show left-sided expression in the lateral plate mesoderm and brain region. We have isolated the nodal-related gene, CyNodal, from Cynops pyrrhogaster. CyNodal mRNA is detected at the marginal zone and left side of several tissues. The left-sideness of CyNodal mRNA expression is highly conserved throughout vertebrate evolution. However, CyNodal mRNA expression shows little variation from the Xenopus nodal-related gene 1, in that CyNodal gene expression in the left lateral plate mesoderm shifts from posterior to anterior at least twice. PMID- 16377259 TI - Expression of the protein kinase D (PKD) family during mouse embryogenesis. AB - The serine/threonine protein kinase D (PKD) family comprises of three members, PKD1 (PKCmu), PKD2 and PKD3 (PKCnu). Like the related C-type protein kinases (PKCs), PKDs are activated by diacylglycerol (DAG). PKDs have been implicated in numerous intracellular signaling pathways including vesicular transport, cell proliferation, survival, migration and immune responses. While experimental data on this recently discovered kinase family are starting to accumulate family member specific information is still sparse and only small effort has been taken to functionally differentiate the three PKDs. To address this issue we followed the expression patterns of PKD1, 2 and 3 during the development of the mouse embryo. Using specific probe sets for RT-PCR and in situ hybridization, we demonstrate shared and differential expression domains for the three PKD family members in both neuronal and non-neuronal tissues. PMID- 16377260 TI - Calpain 10 and development of diabetes mellitus in cystic fibrosis. AB - Diabetes mellitus (DM) and abnormal glucose tolerance (IGT) are common in cystic fibrosis (CF). The loss of pancreatic beta-cells due to pancreatic fibrosis is thought to be one of the principal causes of diabetes in CF, but the aetiology of DM remains somewhat puzzling. Genetic factors may contribute to the development of CF related diabetes (CFRD). The purpose of this study was to investigate the role of polymorphisms in six genes on IGT or DM incidence. PCR and dHPLC were used to screen DNA samples for polymorphisms. Using 2-h oral glucose tolerance tests, 163 adult pancreatic insufficient CF patients have been subdivided in 3 groups: 54 NGT (normal glucose tolerance), 33 IGT and 76 CFRD. We found the first evidence for the association between CFRD and UCSNP-19 polymorphism in the CAPN10 gene. The UCSNP-19 genotype distribution differed significantly between NGT, IGT and CFRD groups. The difference reflected an increase in the 22 genotype (3 copies of 32-bp sequence) in IGT and CFRD patients (p=0.05). Odds ratio for the homozygote 22 versus homozygote 11 was 3.4 (p=0.02). All allele and genotype distributions for the other polymorphisms were similar in the three groups. In conclusion, our observations suggest that UCSNP-19 of CAPN10 may be involved in the pathogenesis of diabetes in CF. PMID- 16377261 TI - Mechanical limitation during CO2 rebreathing in young patients with cystic fibrosis. AB - The aim of the study was to determine whether a decrease in the ventilatory response to carbon dioxide (CO2) in children with cystic fibrosis (CF) is related to a mechanical limitation of the respiratory muscle capacity. The ventilatory response during CO2 rebreathing was performed in 15 patients (mean forced expiratory volume in 1 s (FEV1): 37 +/- 21% predicted, mean arterial CO2: 41+/- 5 mmHg). The slope of the minute ventilation normalised for weight per mmHg CO2 increment correlated negatively with respiratory muscle output, assessed by the oesophageal (p = 0.002), the diaphragmatic pressure time product (p = 0.01), and the tension time index (p = 0.005). In addition, this slope was correlated with dynamic lung compliance (p < 0.0001) and FEV1 (p = 0.03) but not with airway resistance and maximal transdiaphragmatic pressure. Therefore, an excessive load imposed on the respiratory muscles explains the blunting of the ventilatory response to CO2 in young patients with CF. PMID- 16377263 TI - Heterogeneous pulmonary blood flow in response to hypoxia: a risk factor for high altitude pulmonary edema? AB - High altitude pulmonary edema (HAPE) is a rapidly reversible hydrostatic edema that occurs in individuals who travel to high altitude. The difficulties associated with making physiologic measurements in humans who are ill or at high altitude, along with the idiosyncratic nature of the disease and lack of appropriate animal models, has meant that our understanding of the mechanism of HAPE is incomplete, despite considerable effort. Bronchoalveolar lavage studies at altitude in HAPE-susceptible subjects have shown that mechanical stress related damage to the pulmonary blood gas barrier likely precedes the development of edema. Although HAPE-susceptible individuals have increased pulmonary arterial pressure in hypoxia, how this high pressure is transmitted to the capillaries has been uncertain. Using functional magnetic resonance imaging of pulmonary blood flow, we have been able to show that regional pulmonary blood flow in HAPE susceptible subjects becomes more heterogeneous when they are exposed to normobaric hypoxia. This is not observed in individuals who have not had HAPE, providing novel data supporting earlier suggestions by Hultgren that uneven hypoxic pulmonary vasoconstriction is an important feature of those who develop HAPE. This brief review discusses how uneven hypoxic pulmonary vasoconstriction increases regional pulmonary capillary pressure leading to stress failure of pulmonary capillaries and HAPE. We hypothesize that, in addition to the well documented increase in pulmonary vascular pressure in HAPE-susceptible individuals, increased perfusion heterogeneity in hypoxia results in lung regions that are vulnerable to increased mechanical stress. PMID- 16377262 TI - Mechanisms of mucin production by rhinovirus infection in cultured human airway epithelial cells. AB - Mucus hypersecretion relates to exacerbations of bronchial asthma and chronic obstructive pulmonary disease (COPD) caused by rhinovirus (RV) infection. We examined the mechanisms of RV infection-induced mucin production in human tracheal surface epithelial cells and submucosal gland cells. RV14 up-regulated the mRNA expression of MUC2, MUC3, MUC5AC, MUC5B and MUC6, and increased MUC5AC and total mucin concentration in supernatants and lysates of the surface cells. An inhibitor of the nuclear factor kappaB caffeic acid phenylethyl ester, inhibitors of selective p44/42 mitogen-activated protein kinase-kinase PD98059 and U0126, and a selective Src inhibitor PP1 attenuated MUC5AC mRNA expression, and secretion and production of MUC5AC and total mucin glycoprotein in the surface cells. In the gland cells, RV14 also increased mRNA expression of MUC2, MUC5AC, MUC5B and MUC7, and the inhibitors attenuated the secretion of total mucin glycoprotein. Src-related p44/42 mitogen-activated protein kinase pathway may be associated with RV-induced mucin hypersecretion in human airways. PMID- 16377264 TI - Measuring environmental phenols and chlorinated organic chemicals in breast milk using automated on-line column-switching-high performance liquid chromatography isotope dilution tandem mass spectrometry. AB - Breast milk is one possible route of exposure to environmental chemicals, including phenols and chlorinated organic chemicals for breast-fed infants. We developed a highly sensitive method of analyzing breast milk for triclocarban (3,4,4'-trichlorocarbanilide) and eight phenolic compounds: bisphenol A (BPA), 4 tert-octylphenol (4-tOP), ortho-phenylphenol (OPP), 2,4-dichlorophenol, 2,5 dichlorophenol, 2,4,5-trichlorophenol, 2,4,6-trichlorophenol, and 2-hydroxy-4 metoxybenzophenone (BP-3). The method includes adding a solution containing a stable isotope of each chemical, enzymatic hydrolysis of the conjugated chemicals in the milk, and on-line solid-phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry. It can also be used to measure the free (unconjugated) species by omitting the enzymatic deconjugation step. The method, validated using pooled breast milk samples, has inter-day coefficient of variations ranging from 4.8 to 18.9% for most analytes, and spiked recoveries generally about 100%. Detection limits for most analytes are below 1 ng/mL in 100 microL of breast milk. We tested the usefulness of the method by measuring concentrations of these nine compounds in 20 breast milk samples. BPA, OPP, and BP-3 were detected in more than 60% of the samples tested. The free species of these compounds appear to be most prevalent in milk. PMID- 16377265 TI - Height, weight and earnings among coalminers in India. AB - This paper analyses earning/wage differentials by height among coalmine workers in India. Our findings suggest that workers of above average height earn 9-17% more than their shorter counterparts and 6-13% more than average reference height. The results suggest that long-term investments in health human capital might ensure increase of labour productivity and thereby earnings, particularly in underdeveloped economies. PMID- 16377266 TI - Local protein flexibility as a prerequisite for reversible chromophore isomerization in alpha-phycoerythrocyanin. AB - Phycoerythrocyanin is the only cyanobacterial phycobiliprotein containing phycoviolobilin as a chromophore. The phycoviolobilin chromophore is photo reactive; upon irradiation, the chromophore undergoes a Z/E-isomerization involving the rotation of pyrrole-ring D. We have determined the structure of trimeric phycoerythrocyanin at three different experimental settings: monochromatically at 110 K and 295 K as well as with the Laue method at 288 K. Based on their chemical structures, the restraints for the phycoviolobilin of the alpha-subunit and for the phycocyanobilin chromophores of the beta-subunit were newly generated, which allows a chemically meaningful refinement of both chromophores. All three phycoerythrocyanin structures are very similar; the subunits match within 0.5 A. The detailed comparison of the data obtained with the different measurements provided information about the protein properties around the phycoviolobilin chromophore. For the first time, crystals of a phycobilisome protein are used successfully with the Laue technique. This paves the way for time-resolved macromolecular crystallography, which is able to elucidate the exact mechanisms of the phycoviolobilin photoactivity including the protein involvement. PMID- 16377267 TI - Elimination of HIV infection in infants in Europe--challenges and demand for response. AB - Effective interventions for the prevention of mother-to-child transmission (MTCT) of human immunodeficiency virus (HIV) infection now exist and where these are fully implemented, MTCT rates of 1-2% are achievable. Virtual elimination of HIV in infants by 2010 has therefore been set as a goal for European region. There are, however, many challenges. The eastern European and central Asian countries are facing the fastest growing HIV epidemic in the world with a dramatic increase in numbers of HIV-positive pregnancies and new HIV infected infants. Nevertheless, the prevalence of HIV among pregnant women is still relatively low and the high coverage with antenatal care provide an opportunity to decrease the number of new HIV cases among infants to minimal level. The challenge is to move national strategies for prevention of HIV infection among infants from a disease focused vertical approach towards effective preventive interventions integrated into mother-child health (MCH) and reproductive health services. The scaling up of prevention interventions among those most at risk and hard-to-reach women are key priority actions. This goes beyond clinical care and needs to include a range of care and protection issues, both in health institutions and in the community. The WHO Regional Office for Europe, together with other UNAIDS co-sponsors, has developed a regional strategic framework for prevention of HIV infection in infants. The strategic framework promotes a comprehensive approach comprising the four interrelated elements: (1) primary prevention of HIV infection; (2) prevention of unintended pregnancies among HIV-infected women; (3) prevention of HIV transmission from HIV-infected women to their children; (4) provision of care and support to HIV-infected women, their infants and families. Implementation of all four pillars of the strategic framework would help European countries to achieve the goal of virtual elimination of HIV infection in infants. PMID- 16377268 TI - Type II/III Runx2/Cbfa1 is required for tooth germ development. AB - Runx2/Cbfa1 is an essential transcription factor for osteoblast differentiation and bone formation. Runx2/Cbfa1 knockout mice showed both a complete lack of ossification and the developmental arrest of tooth germ. We here report Runx2/Cbfa1 isoform-type specific functional roles in the development of tooth germ by the administration of antisense phosphorothioate oligodioxynucleotides (S ODNs) into cultured mouse mandibles. The administration of type II/III Runx2/Cbfa1 antisense S-ODNs into the culture media resulted in an arrest of tooth germ growth at the bud-like stage in cultured mandible taken from the 11 day-old embryos, while also causing the inhibition of the differentiation of odontogenic cells into ameloblast and odontoblast in cultured tooth germs taken from the 15-day-old embryos. The expression of dentin matrix protein 1, dentin sialophosphoprotein, amelogenin, and ameloblastin was shown to be markedly suppressed in cultured tooth germ by the semi-quantitative RT-PCR. Meanwhile, no developmental arrest of tooth germ, no inhibition of gene expression, or differentiation of odontogenic cells was observed in samples treated with the type I Runx2/Cbfa1 antisense S-ODNs. The same findings were also observed in either the control or the sense and random sequence S-ODNs-treated samples. These data indicate that the type II/III Runx2/Cbfa1 isoform is closely related to the development and differentiation of tooth germ. PMID- 16377269 TI - The calcium-sensing receptor regulates PTHrP production and calcium transport in the lactating mammary gland. AB - Lactating mammals must supply large amounts of calcium to the mammary gland where it is transported across mammary epithelial cells and into milk. This demand for calcium is associated with transient loss of bone mass, triggered, in part, by the secretion of parathyroid hormone-related protein (PTHrP) from the mammary gland into the circulation. The calcium-sensing receptor (CaR) is a G-protein coupled receptor that signals in response to extracellular calcium ions. It is responsible for coordinating calcium homeostasis by regulating parathyroid hormone secretion in the parathyroid glands and by regulating calcium handling in the renal tubules. Previous studies had shown that the CaR is expressed on mammary epithelial cells during lactation, and it had been suggested that CaR signaling in the mammary gland helps to coordinate its production of PTHrP and calcium transport into milk. In this study, we examined mammary gland PTHrP production and calcium transport in CaR(+/-) mice, a genetic model of CaR insufficiency. We found that haploinsufficiency for the CaR resulted in increased PTHrP production both in vivo and in vitro. In contrast, CaR haploinsufficiency impaired calcium transport into milk in vivo and transepithelial calcium transport by mammary epithelial cells in vitro. These data provide genetic confirmation that the CaR regulates PTHrP production and calcium transport in the lactating mammary gland. This allows the mammary gland to become a calcium sensing organ and to participate in systemic calcium homeostasis during lactation. PMID- 16377270 TI - Potential predictive factors of osteoporosis in HIV-positive subjects. AB - Recent reports showed a high frequency of osteopenia/osteoporosis in HIV-infected subjects. Mechanism on the basis of this alteration is still unclear, as the direct effect of virus or of antiretroviral drugs. One hundred sixty-one consecutive HIV-infected outpatients aged 30-50 years, both naive and HAART treated for >1 year, were included. An interview questionnaire was performed to establish prior pathological, toxic, epidemiological histories, medications intake, physical activity and eating habits. Blood and urinary tests were checked to exclude concomitant diseases, as were markers of bone metabolism and vitamin D3-metabolites. Each subject underwent to a lumbar spine and left hipbone mineral density by DEXA, using WHO criteria for diagnosis of osteopenia/osteoporosis. Radiologist was unaware if the subject was receiving HAART or not. For groups' homogeneity Chi-square, Fisher's exact and Student's t tests were used. Logistic regression analysis was used to find predictors of osteopenia/osteoporosis and linear regression model to find differences in bone mass density. The demographic characteristics of the 48 naive subjects and the 113 on HAART were comparable. Eighty subjects (49.7%) showed osteopenia/osteoporosis: 22 (45.8%) naive and 58 (51.3%) on HAART (P = 0.46). Independent predictors of osteopenia/osteoporosis were female gender (OR: 3.02, 95% CI: 1.26-7.25, P = 0.01 vs. male), older age (OR: 1.10, 95% CI: 1.01-1.20, P = 0.03, for each additional year), low body mass index (OR: 0.78, 95% CI: 0.68-0.91, P = 0.001 for each additional unit) and higher HIV-RNA levels at DEXA (OR: 1.97, 95% CI: 1.16-3.34, P = 0.01 for each additional Log(10)), whereas the use of HAART (OR: 2.61, 95% CI: 0.66-10.27, P = 0.17 vs. naive) and the alterations of markers of bone metabolism were not significantly related to osteopenia/osteoporosis. Similar findings were obtained using linear regression model analysis. HIV-infected subjects have a high frequency of osteopenia/osteoporosis. Traditional risk factors are predictive of osteopenia/osteoporosis also in HIV-subjects; the association with higher HIV-RNA levels can suggest a direct role of HIV itself in the occurrence of bone disease. PMID- 16377271 TI - Role of macrophages in LPS-induced osteoblast and PDL cell apoptosis. AB - In periradicular lesions and periodontal disease, bacterial invasion leads to chronic inflammation resulting in disruption of the structural integrity of the periodontal ligament and progressive alveolar bone destruction. The pathogenesis of these conditions has been attributed not only to bacterial-induced tissue destruction but also to a defect in periodontal tissue repair. Accumulated data have also shown that lipopolysaccharide (LPS) can directly induce cell death or apoptosis in many cell types, including macrophages, osteoblasts, vascular endothelial cells, hepatocytes and myocytes. The present study hypothesized that bacterial LPS-induced apoptosis in osteoblasts and periodontal ligament fibroblasts (PDL cells) is an important contributing factor to the defect in periodontal tissue repair in periodontal and periapical disease. Macrophages have been shown to respond to bacterial LPS by increasing the production of proinflammatory cytokines. In addition, large numbers of macrophages are present in inflamed periodontal tissue. We speculated that macrophages were a potential candidate cell for mediating apoptosis in osteoblasts and PDL cells in response to bacteria-derived LPS. The macrophage-like cell line, RAW 264.7, was stimulated with LPS, and the conditioned medium was used to treat osteoblasts and PDL cells. Bacterial LPS had no direct apoptotic effect on mouse osteoblasts or PDL cells, whereas the conditioned medium from LPS-activated macrophages was able to induce apoptosis in these cells. To evaluate the contribution of tumor necrosis factor alpha (TNF-alpha) released from macrophages on osteoblast and PDL cell apoptosis, cells were incubated with conditioned medium from LPS-treated macrophages in the presence and absence of anti-TNF-alpha neutralizing antibodies. TNF-alpha neutralizing antibody pretreatment inhibited the effect of conditioned medium from LPS-treated macrophages on osteoblast and PDL cell apoptosis in a dose dependent manner. These results suggest that LPS could indirectly induce apoptosis in osteoblasts and PDL cells through the induction of TNF-alpha release from macrophages. These studies provide insight into a potential mechanism by which bacterial-derived LPS could contribute to defective periodontal and bone tissue repair in periodontal and periapical disease. PMID- 16377272 TI - Prognostic implications of a normal stress technetium-99m-tetrofosmin myocardial perfusion study in patients with a healed myocardial infarct and/or previous coronary revascularization. AB - Previous studies have shown a good outcome for patients who present with normal findings on stress myocardial perfusion imaging. Currently, the prognostic implications of a normal study in patients who have a history of coronary artery disease (CAD) are not clear. This study investigated the long-term prognosis after a normal finding on stress technetium-99m (Tc-99m)-tetrofosmin single photon emission computed tomography in patients with a history of CAD. The study included 147 consecutive patients with a history of CAD (previous myocardial infarction and/or myocardial revascularization), who underwent exercise bicycle or high-dose dobutamine-atropine stress Tc-99m-tetrofosmin single-photon emission computed tomography, and had normal perfusion results during stress and at rest. Follow-up was completed in all patients. During a follow-up of 6.5 +/- 1.9 years, 20 patients (14%) died, 10 (7%) of whom died due to cardiac causes, and 12 (8%) had a nonfatal myocardial infarction. Annual cardiac death rates were 0.5% during the first 3 years of follow-up and 1.3% in the subsequent 3 years. Independent predictors of cardiac death were male gender, rate-pressure product at rest, and rate-pressure product at peak stress. In conclusion, patients who have a history of CAD have a very low cardiac death rate during the 3 years after a normal finding on stress Tc-99m-tetrofosmin single-photon emission computed tomography. Repeated testing should be reconsidered 3 years after the initial evaluation and when a change in symptoms or clinical condition occurs. PMID- 16377273 TI - Infarct-related coronary artery patency and medication use prior to ST-segment elevation myocardial infarction. AB - In patients who have ST-segment elevation myocardial infarction (STEMI), a patent infarct-related artery on the initial angiogram is associated with improved clinical outcomes, including decreased mortality. The present study assessed the influence of administering aspirin, beta blockers, statins, and angiotensin converting enzyme inhibitors before STEMI on infarct-related artery patency. Our data demonstrate that patients who have STEMI and receive these medications on an outpatient basis before the event have a higher likelihood of having a patent infarct-related artery compared with patients who do not receive these medications. Further, our data demonstrate a graded association according to the number of such medications being administered: the likelihood of a patent infarct related artery increased to >50% as the number of these medications increased. PMID- 16377274 TI - Relation of nocturnal melatonin levels to C-reactive protein concentration in patients with ST-segment elevation myocardial infarction. AB - C-reactive protein, a marker of inflammation, is increasingly measured to stratify risk for coronary artery disease and guide clinical management. Likewise, a relation has been suggested between decreased nocturnal melatonin levels and coronary artery disease. However, little is known about the association between melatonin and C-reactive protein in patients with ischemic heart disease. PMID- 16377275 TI - Remodeling characteristics of minimally diseased coronary arteries are consistent along the length of the artery. AB - Using a method that creates anatomically correct, 3-dimensional arterial reconstructions, 55 minimally diseased coronary arteries from 40 patients were studied. Homogenous remodeling characteristics along the entire length of the artery were observed in 48 arteries (87%). In the aggregate, arteries exhibited compensatory expansive remodeling. Individually, the full spectrum of compensatory expansive remodeling (60%), excessive expansive remodeling (21%), and constrictive remodeling (19%) was observed across arteries. Each artery was consistent in its remodeling characteristics from proximal to distal portions of the artery, and the remodeling pattern of each artery was independent within the same patient. PMID- 16377276 TI - Two-year clinical outcomes after enhanced external counterpulsation (EECP) therapy in patients with refractory angina pectoris and left ventricular dysfunction (report from The International EECP Patient Registry). AB - Enhanced external counterpulsation (EECP) is a noninvasive circulatory assist device that has recently emerged as a treatment option for refractory angina in left ventricular (LV) dysfunction. This 2-year cohort study describes the long term follow-up of patients who had severe LV dysfunction that was treated with EECP for angina pectoris and reports clinical outcomes, event-free survival rates, and the incidence of repeat EECP. This study included 363 patients who had refractory angina and LV ejection fraction < or =35%. Most patients reported quality of life as poor. After completion of treatment, there was a significant decrease in severity of angina class (p < 0.001), and 72% improved from severe angina to no angina or mild angina. Fifty-two percent of patients discontinued nitroglycerin use. Quality of life improved substantially. At 2 years this decrease in angina was maintained in 55% of patients. The 2-year survival rate was 83%, and the major adverse cardiovascular event-free survival rate was 70%. Forty-three percent had no reported cardiac hospitalization; 81% had no reported congestive heart failure events. Repeat EECP was performed in 20% of these patients. The only significant independent predictor of repeat EECP in a proportional hazard model was failure to complete the first EECP treatment course (hazard ratio 2.9, 95% confidence interval 1.7 to 4.9). Improvements in angina symptoms and quality of life were maintained at 2 years. In conclusion, for patients who have high-risk LV dysfunction, EECP offers an effective, durable therapeutic approach for refractory angina. Decreased angina and improvement in quality of life were maintained at 2 years, with modest repeat EECP and low major cardiovascular event rates. PMID- 16377277 TI - Serial angioscopic findings after successful intracoronary brachytherapy for in stent restenosis. AB - The intracoronary changes that occur after brachytherapy for in-stent restenosis (ISR) have yet to be fully established. The purpose of this study in patients who had ISR was to examine the serial angioscopic changes in intracoronary lesions that occurred after brachytherapy. Forty-four patients who had ISR (49 lesions) underwent balloon angioplasty (n = 34) or directional atherectomy (n = 15), followed by intracoronary brachytherapy using a beta-emitting phosphorus-32 source wire. Angioscopic investigations were performed 3 and 9 months after brachytherapy. Uncovered stents were detected in 63.3% of lesions at 3 months. A significant decrease (p = 0.028) in this prevalence occurred over the next 6 months, with 36% of lesions having uncovered stents at 9 months. At 3 months, 33% of the lesions had visible erosion or ulceration and superficial thrombus. The prevalence of these characteristics was decreased at 9 months, although 17% of the lesions were still ulcerated or eroded at that time. Protruding thrombus was not observed in any lesion at 3 and 9 months. In conclusion, uncovered stents and intimal erosions or ulcerations were still present 9 months after brachytherapy in 36% and 17% of lesions, respectively. These results suggest that the healing process was not completed 9 months after brachytherapy in approximately 33% of lesions. PMID- 16377278 TI - Detection of von Willebrand factor and tissue factor in platelets-fibrin rich coronary thrombi in acute myocardial infarction. AB - The rapid closure of coronary arteries due to occlusive thrombi is the major cause of acute myocardial infarction. However, the mechanisms of coronary thrombus formation have not been elucidated. We immunohistochemically assessed the localizations and their changes over time of glycoprotein IIb/IIIa, fibrin, von Willebrand factor (vWF), and tissue factor (TF), after the onset of chest pain (<4, 4 to 6, or 6 to 12 hours), in fresh coronary thrombi causing acute myocardial infarction. The occlusive thrombi were consistently composed of platelets, fibrin, vWF, and TF from the early phase of onset, and glycoprotein IIb/IIIa and fibrin were closely associated with vWF and TF, respectively. vWF and/or TF may contribute to occlusive thrombus formation and be novel therapeutic candidates for treating patients with coronary thrombosis. PMID- 16377279 TI - Comparison of coronary arterial finding by intravascular ultrasound in patients with "transient no-reflow" versus "reflow" during percutaneous coronary intervention in acute coronary syndrome. AB - Previous studies have shown that transient no-reflow during coronary intervention but with Thrombosis in Myocardial Infarction (TIMI) grade 3 flow at the completion of the procedure is associated with increased in-hospital and 6-month mortality. We hypothesized that the use of intravascular ultrasound before intervention could identify morphologic features that were predictive of transient no-reflow in patients who had acute coronary syndrome (ACS). We analyzed 220 patients with ACS who had suitable intravascular ultrasound images that were acquired before intervention. We defined "transient no-reflow" as TIMI grade 0, 1, or 2 flow during the procedure and TIMI grade 3 flow at the completion of the procedure. We defined "reflow" as good coronary flow (TIMI grade 3 flow) during and after the procedure. Patients were categorized to a transient no-reflow group (n = 20) or a reflow group (n = 200). In the transient no-reflow group, vessel area and amount of plaque burden in the culprit lesion were significantly greater than in the reflow group (vessel 20.8 +/- 5.4 vs 16.4 +/- 6.2 mm(2), p < 0.01; plaque burden 0.90 +/- 0.03 vs 0.83 +/- 0.08, p < 0.001). The presence of ruptured plaque, lipid pool-like images, and thrombus formation were significantly higher in the transient no-reflow group than in the reflow group. Multivariate analysis identified the presence of thrombus formation (odds ratio 4.53, 95% confidence interval 1.03 to 20.0, p = 0.04) and larger plaque burden (odds ratio 1.79, 95% confidence interval 1.01 to 3.23, p = 0.05) as independent predictors of transient no-reflow. In conclusion, lesion morphologies are different for transient no-reflow and reflow. These findings suggest that the presence of thrombus formation and large plaque burden increase the risk for developing transient no-reflow during coronary intervention for ACS. PMID- 16377280 TI - Efficacy of sirolimus-eluting stents compared with bare metal stents for saphenous vein graft intervention. AB - Saphenous vein graft (SVG) intervention is associated with a significantly increased rate of periprocedural complications and late clinical and angiographic restenosis. We examined the efficacy and safety of sirolimus-eluting stents (SESs; Cypher) compared with bare metal stents (BMSs) in SVG intervention. Forty eight patients who had 50 SVG lesions and underwent standard percutaneous coronary intervention with SESs (SES group) were compared with 57 patients who had 64 SVG lesions and underwent intervention with BMSs (BMS group). All patients received distal protection devices during SVG intervention. In-hospital, 30-day, 6-month, and 1-year clinical outcomes in the 2 groups were compared. Baseline clinical and procedural characteristics were balanced between groups. There were no deaths or Q-wave myocardial infarctions during the index hospitalization, but compared with the BMS group, patients in the SES group had significantly fewer non-Q-wave myocardial infarctions (4% vs 21%, p = 0.01), which was mainly attributed to increased periprocedural creatine kinase-MB levels. At 30-day, 6 month, and 1-year follow-ups, all clinical outcomes were similar between groups. Event-free survival at 1 year was also similar between groups (p = 0.84). In conclusion, the use of SESs in patients who undergo SVG intervention with a distal protection device is clinically safe and feasible but is not associated with decreased clinical events up to 1 year compared with BMSs. PMID- 16377281 TI - Influence of aspirin resistance on platelet function profiles in patients on long term aspirin and clopidogrel after percutaneous coronary intervention. AB - Increased platelet inhibition is achieved when clopidogrel is added to aspirin (acetylsalicylic acid [ASA]). A broad variability in platelet inhibition profiles during the early phases of treatment has been demonstrated and may be attributed to ASA resistance. However, the influence of ASA sensitivity on platelet function profiles of patients on long-term dual antiplatelet therapy has yet to be explored. A total of 135 patients who had previously undergone percutaneous coronary intervention on long-term (>1 month) ASA and clopidogrel therapy was included. The PFA-100 system was used to define ASA resistance. Platelet aggregation, after adenosine diphosphate (6 and 20 micromol/L) and collagen (6 microg/ml) stimuli, and platelet activation (glycoprotein IIb/IIIa activation and P-selectin expression), after adenosine diphosphate (2 micromol/L) and thrombin receptor-activating peptide (50 micromol/L) stimuli, were assessed by light transmittance aggregometry and flow cytometry, respectively. Patient variability in response to treatment was defined by the coefficient of variability. ASA resistance was found in 60 of 135 patients (44%). Patients with diabetes were more frequently ASA resistant. Collagen/epinephrine- and collagen/adenosine diphosphate-coated cartridges on the PFA-100 had shorter closure times in the ASA resistant population compared with ASA-sensitive patients. Platelet aggregation and activation were significantly higher in ASA-resistant patients. A broad variability (coefficient of variation >0.25) in patient response to treatment was observed in ASA-resistant and -sensitive patients. In conclusion, ASA resistance is associated with increased platelet reactivity in patients on long-term dual antiplatelet treatment. PMID- 16377282 TI - Effects of morning versus evening intake of atorvastatin on major cardiac event and restenosis rates in patients undergoing first elective percutaneous coronary intervention. AB - It is not known whether taking atorvastatin in the morning versus in the evening has a different effect on major cardiac event and restenosis rates in patients undergoing percutaneous coronary intervention. Therefore, the aim of the present study was to investigate the effects of morning versus evening intake of atorvastatin on major cardiac events and restenosis rates and also on serum lipid and high sensitivity C-reactive protein levels in patients with single-vessel disease who underwent first elective percutaneous coronary intervention. PMID- 16377283 TI - ST-segment analysis to predict infarct size and functional outcome in acute myocardial infarction treated with primary coronary intervention and adjunctive abciximab therapy. AB - ST-segment resolution is used to classify the response to reperfusion therapy in acute myocardial infarction, but the possibility to predict outcome in individual patients is unclear, particularly in the setting of primary percutaneous coronary intervention (PCI) and abciximab therapy. We studied 213 patients who underwent successful revascularization with PCI. Maximal ST-segment elevation was measured before and 30 minutes after PCI. Patient outcome was defined on the basis of infarct size and left ventricular ejection fraction (EF) as derived from gated single-photon emission computed tomography that was acquired 1 month after infarction. Patients who had > or =50% ST resolution showed a smaller infarct (15.1 +/- 13.6% vs 19.9 +/- 15.7%, p < 0.05) but not a higher left ventricular EF (48.7 +/- 12.3% vs 45.2 +/- 11.8%) than did patients who had <50% resolution. According to cluster analysis of infarct size and left ventricular EF, 132 patients had favorable outcome (central values: infarct size 7.5%, left ventricular EF 55%) and 81 did not (central values: infarct size 30%, left ventricular EF 36%). Using receiver-operating characteristic curve analysis, the optimal ST-resolution cutoff was >60%, with 77% sensitivity and 51% specificity for predicting favorable outcome. ST-segment elevation < or =4.5 mV before PCI was 80% sensitive and 48% specific, and ST-segment elevation < or =1 mV after PCI was 74% sensitive and 60% specific for predicting favorable outcome. In conclusion, in the setting of primary PCI and abciximab therapy, ST-segment elevation resolution requires a high threshold (>60%) to effectively classify patients; the capability of ST-segment analysis to predict patient outcome is limited, with ST-segment elevation after PCI showing the best compromise between sensitivity and specificity. PMID- 16377284 TI - Statins and postoperative risk of atrial fibrillation following coronary artery bypass grafting. AB - Atrial fibrillation (AF) is a common complication after coronary artery bypass grafting. Atrial remodeling has been observed in AF and has been associated with the development of this arrhythmia. Because 3-hydroxy-3-methylglutaryl coenzyme A inhibitors (statins) have been demonstrated to modify remodeling, we hypothesized a protective role of statins against postoperative AF. We also hypothesized that extracellular matrix turnover and brain natriuretic peptide (BNP) might be related to such atrial remodeling. We studied 234 consecutive patients who underwent coronary artery bypass grafting (173 men; 65 +/- 9 years of age) in whom the occurrence of postoperative AF was monitored. In a subgroup of 66 patients, we measured plasma levels of matrix metalloproteinase-1 (MMP-1), its inhibitor, tissue inhibitor matrix metalloproteinase-1 (TIMP-1; as indexes of extracellular matrix remodeling), and N-terminus pro-BNP (related to left ventricular function) at baseline and at 24 hours after surgery. Of 234 patients, 66 (28.2%) developed postoperative AF. In multivariate analysis, previous AF was related to an increase in the development of AF (odds ratio 11.92, 95% confidence interval 2.37 to 59.98, p = 0.026), whereas statin use was related to a decrease in arrhythmia (odds ratio 0.52, 95% confidence interval 0.28 to 0.96, p = 0.038). A higher TIMP-1/MMP-1 ratio at 24 hours after surgery was present in those who did not develop postoperative AF (p = 0.043). Statin use was associated with increased TIMP-1 levels and TIMP-1/MMP-1 ratio (p = 0.027 and 0.036, respectively). No significant relations to N-terminus pro-BNP were seen. In conclusion, previous AF and nonuse of statins are significantly associated with AF after coronary artery bypass grafting. Statin use may be protective against AF after coronary artery bypass grafting, possibly due to alterations in the extracellular matrix and remodeling after coronary artery bypass grafting. PMID- 16377285 TI - Comparative safety of atorvastatin 80 mg versus 10 mg derived from analysis of 49 completed trials in 14,236 patients. AB - Atorvastatin has been shown to reduce coronary events and revascularization procedures in patients with multiple risk factors for coronary heart disease. Recent studies with atorvastatin 80 mg support the overall safety of this dose during long-term treatment. However, physicians appear reluctant to use high doses of statins. A retrospective analysis of pooled data from 49 clinical trials of atorvastatin in 14,236 patients treated for an average period of 2 weeks to 52 months was conducted. The study compared the safety of atorvastatin 10 mg (n = 7,258), atorvastatin 80 mg (n = 4,798), and placebo (n = 2,180) and included analyses on treatment-associated adverse events; nonserious and serious adverse events related to the musculoskeletal, hepatic, and renal systems; the incidence of elevations of creatine kinase >10 times the upper limit of normal (ULN); and hepatic transaminases >3 times ULN. Percentages of patients experiencing > or =1 adverse event were similar across all 3 groups. Withdrawals due to treatment related adverse events were observed in 2.4%, 1.8%, and 1.2% of patients in the atorvastatin 10 mg, atorvastatin 80 mg, and placebo groups, respectively. Serious adverse events were rare and seldom led to treatment withdrawal with any dose. Treatment-associated myalgia was observed in 1.4%, 1.5%, and 0.7% of patients in the atorvastatin 10 mg, atorvastatin 80 mg, and placebo groups, respectively. No cases of rhabdomyolysis were reported in any group. Persistent elevations in hepatic transaminases >3 times ULN were observed in 0.1%, 0.6%, and 0.2% of patients in the atorvastatin 10 mg, atorvastatin 80 mg, and placebo groups, respectively. The incidence of treatment-associated adverse events for atorvastatin 80 mg was similar to that of atorvastatin 10 mg and placebo. In conclusion, the results of this analysis support the positive safety profile of atorvastatin at the highest dose. PMID- 16377286 TI - Association of increased body mass index and impaired endothelial function among Hispanic women. AB - We tested the hypothesis that an increased body mass index was similarly associated with impaired endothelial function as measured by flow-mediated dilation in a high-risk, Hispanic population of men and women living in northern Manhattan. The association of flow-mediated dilation and body mass index was significant in women (beta -0.16 +/- 0.04, p <0.0001) but not in men (beta -0.02 +/- 0.06, p = 0.72). This is the first study to demonstrate a gender-specific difference in endothelial function associated with body mass index. PMID- 16377287 TI - Effect of glycemic control on left ventricular diastolic function in type 1 diabetes mellitus. AB - Left ventricular (LV) diastolic dysfunction is a main feature of diabetic heart disease. The aim of this prospective study was to evaluate the influence of glycemic control on diastolic function in type 1 diabetes mellitus. Thirty-six normotensive (24-hour blood pressure <130/80 mm Hg) subjects with inadequately controlled (glycated hemoglobin >7%) type 1 diabetes, without clinically detectable heart disease, were enrolled. After the basal evaluation, insulin therapy was modified to improve glycemic control. Glycated hemoglobin, LV echocardiography, 24-hour blood pressure monitoring, and laboratory tests were repeated after 6 months in all patients and after 12 months in 27 patients. At the basal evaluation, LV anatomy and systolic function were normal in all, and diastolic function was impaired in 14 patients. After 6 months, the mean values of body mass index, 24-hour blood pressure, and LV anatomy and systolic function were unchanged; mean glycated hemoglobin was decreased (p < 0.001), and mean values of diastolic parameters were significantly improved. After 12 months, the mean values of all blood pressure, metabolic, and LV parameters were unchanged. Percent changes of diastolic parameters were inversely correlated with percent changes of glycated hemoglobin, considering changes from the basal to the 6-month evaluation, as well as changes from the 6- to the 12-month evaluation. In conclusion, in normotensive patients with type 1 diabetes, a close relation was found between glycemic control and LV diastolic function, which improves when glycemic control improves. Therefore, diastolic dysfunction can be prevented or reversed, at least partly, by tight glycemic control. PMID- 16377288 TI - Flow mediated dilatation and cardiac function in type 1 diabetes mellitus. AB - The present study sought to examine the occurrence of subclinical markers of cardiovascular risk and cardiac dysfunction with increasing disease duration in type 1 diabetes mellitus (DM). There are few data on subclinical cardiovascular abnormalities in type 1 DM. The study included 100 patients without any cardiovascular complaints (mean age 46.6 years, range 22 to 63), with a history of type 1 DM ranging from 2 to 36 years, and 75 age-matched controls. Standard 2 dimensional and Doppler echocardiography and pulse-wave tissue Doppler (tD) mapping of systolic (Sm) and diastolic (Em, Am) velocities (12 left ventricular [LV] segments and right-sided cardiac) were performed. An Em/Am ratio of <1 was considered to represent abnormal segmental diastolic function. Flow-mediated dilation (FMD) of the brachial artery, carotid intima media thickness (IMT) measurement, and extensive laboratory analysis were performed. The FMD was reduced, and IMT increased in patients (p < 0.01). Regional tD-derived diastolic and systolic functional abnormalities were observed within the first decade of the disease. Significant correlations were found between FMD and LV segments with tD-derived dysfunction, the duration of DM, and fibrinogen (p < 0.0001 for all). Stepwise regression analysis showed that FMD was the strongest predictor of abnormal segmental function (p < 0.0001). Data further presented as an analysis of tertiles by DM duration show an increasing occurrence of subclinical cardiac dysfunction and cardiovascular risk markers compared with age-matched controls. In conclusion, FMD is associated with abnormal segmental cardiac function in type 1 DM. PMID- 16377289 TI - Left atrial volume, geometry, and function in systolic and diastolic heart failure of persons > or =65 years of age (the cardiovascular health study). AB - The left atrium enlarges in association with many factors, including aging, atrial fibrillation, hypertension, diastolic dysfunction, and heart failure (HF) with low ejection fraction. However, left atrial (LA) volume, geometry, and emptying have not been compared between diastolic and systolic HF, nor has the association of LA volume for new HF been determined in older subjects, many of whom have normal ejection fraction. We used echocardiography to measure the LA volume, geometry, and emptying in 851 community-dwelling subjects > or =65 years of age, including 180 with HF at baseline and 255 participants who subsequently developed HF. The LA volume, area, and linear dimensions were higher in the prevalent and incident HF groups than in controls and did not differ between those with systolic versus diastolic HF, independent of co-morbidities and Doppler measures of diastolic function. The fractional area change was associated with prevalent, but not incident, HF. In conclusion, in population-based older subjects, the LA size is increased and LA emptying decreased in patients with either systolic or diastolic HF. LA size is associated with the new development of HF. These findings highlight the important role of the left atrium in HF, with or without a decreased ejection fraction. PMID- 16377290 TI - Relation of circulating C-reactive protein to progression of aortic valve stenosis. AB - C-reactive protein (CRP) is a marker of inflammation and predicts outcome in apparently healthy subjects and patients with coronary artery disease. Systemic inflammation is present in patients with aortic valve stenosis (AS). The aim of this prospective study was to assess whether CRP levels predict the progression of AS severity. Blood samples for high-sensitivity CRP measurements and echocardiographic data were obtained in 43 patients (70% men; mean age 73 +/- 8 years) with asymptomatic degenerative AS at study entry. On the basis of repeat echocardiographic assessment at 6 months, patients were grouped as (1) slow progressors (a decrease in aortic valve area [AVA] <0.05 cm2 and/or an increase in aortic peak velocity <0.15 m/s) and (2) rapid progressors (a decrease in AVA > or =0.05 cm2 and/or an increase in aortic peak velocity > or =0.15 m/s). Plasma CRP levels were significantly higher in rapid progressors than slow progressors (median 5.1 [range 2.3 to 11.3] vs 2.1 [range 1.0 to 3.1] mg/L, p = 0.007). In multivariate analysis, CRP levels >3 mg/L were independently associated with rapid AS progression (odds ratio 9.1, 95% confidence interval 2.2 to 37.3). In conclusion, CRP levels are higher in patients with degenerative AS who show rapid valve disease progression. These findings suggest that inflammation may have a pathogenic role in degenerative AS. PMID- 16377291 TI - Feasibility and diagnostic accuracy of quantitative assessment of mechanical prostheses leaflet motion by transthoracic and transesophageal echocardiography in suspected prosthetic valve dysfunction. AB - Transthoracic (TTE) and transesophageal echocardiography (TEE) are the standard techniques for the evaluation of prosthetic valve function. However, quantitative evaluation of leaflet(s) motion is not routinally carried out, although leaflet(s) opening and closing angle measurements are important information to recognize prosthetic dysfunction. For this purpose, cinefluoroscopy is considered the "gold standard" technique. The aim of this study was the evaluate the diagnostic accuracy of TTE and TEE in the quantitative assessment of leaflet motion in patients with mechanical protheses. One hundred-eleven patients with mitral (single disk 18; bileaflet 48) and aortic (single disk 22; bileaflet 23) prostheses, were referred to TTE, TEE, and cinefluoroscopy for electrical cardioversion of atrial fibrillation (n = 40) or suspected prosthesis dysfunction (n = 71). Echocardiographic evaluation included leaflet(s) opening and closing angle measurements; results were compared with cinefluoroscopy. For mitral prostheses, opening and closing angles were correctly identified by TTE in 85% and by TEE in 100% of patients, regardless of prosthetic valve type, with a good concordance with cinefluoroscopy. For aortic prostheses, opening angles were correctly identified by TTE and TEE, respectively, in 40% and 77% of patients with single-disk and in 13% and 35% of patients with bileaflet prostheses. Both TTE and TEE were rarely able to identify closing angles. In conclusion, quantitative evaluation of mitral leaflet(s) motion may be accurately achieved with TTE and TEE, leading to increased diagnostic efficacy of prosthetic valve dysfunction. In the aortic position, TTE and TEE allow a quantitative evaluation of leaflet(s) dynamics only in a minority of patients and cinefluoroscopy still remains the first-choice technique. PMID- 16377292 TI - Cardiac ventricular diastolic and systolic duration in children with heart failure secondary to idiopathic dilated cardiomyopathy. AB - Systole and diastole are the fundamental periods of the cardiac cycle, yet little emphasis has been placed on their relative duration when evaluating heart failure. Cardiac intervals are used to assess ventricular function, but the relative duration of systole and diastole for defining function have not been evaluated. We hypothesized that in heart failure, systole is prolonged and diastole shortened. We defined systole and diastole in 16 children with idiopathic dilated cardiomyopathy and in 16 normal controls, matched for age and gender, using the mitral regurgitant (MR) and tricuspid regurgitant (TR) flow duration. The systole and diastole durations (expressed as a fraction of the cardiac cycle) were correlated with heart rate and age and compared between groups. The subjects were compared with gender- and age-matched controls (9.98 +/ 6.1 vs 9.88 +/- 6.08 years, p = NS). The 2 groups had similar heart rates (104 +/- 31 vs 92 +/- 34 beats/min, p = NS). The systole duration was not significantly different when measured by MR versus TR duration (0.60 +/- 0.10 vs 0.57 +/- 0.11 of the cardiac cycle, respectively, p = NS). Systole was prolonged in subjects compared with controls (0.60 +/- 0.1 vs 0.42 +/- 0.08, respectively, using MR duration, p < 0.0001 and 0.57 +/- 0.11 vs 0.41 +/- 0.07, respectively, using TR duration, p = 0.0008). The systolic/diastolic ratio was 0.77 +/- 0.24 in the controls versus 1.57 +/- 0.98 in the patients with idiopathic dilated cardiomyopathy using the TR duration (p < 0.005) and 1.67 +/- 0.68 using the MR duration (p < 0.0001). The systole duration correlated with heart rate in subjects (r = 0.79, p = 0.0003) and controls (r = 0.69, p = 0.003). In conclusion, systole is significantly prolonged and diastole correspondingly shortened in heart failure. Reversal of the normal systolic/diastolic ratio may compromise cardiac filling and function. The systole and diastole duration are easily measured using routine Doppler flow, enhancing assessment of cardiac function in children. PMID- 16377293 TI - Comparison of diastolic septal perforator flow velocities in hypertrophic cardiomyopathy versus hypertensive left ventricular hypertrophy. AB - In this study, we measured diastolic septal perforator flow velocities by Doppler transthoracic echocardiography (TTE) in patients with hypertrophic cardiomyopathy (HCM). Using color-guided pulsed Doppler TTE, septal perforator flow velocity recordings were attempted in 69 patients and successfully recorded in 47 (68%). First, we compared 14 patients with HCM to 12 controls and to 11 patients with hypertension with left ventricular hypertrophy. Next, in 10 additional patients with HCM, we compared the septal velocities with the epicardial left anterior descending artery (LAD) velocities recorded during the same TTE study. In the patients with HCM, the peak septal diastolic velocities were twice that of the normal controls (88 +/- 40 vs 41 +/- 13 cm/s) and also higher than in hypertensive left ventricular hypertrophy (51 +/- 18 cm/s, p < 0.0001). All 10 patients with HCM showed a step-up of peak diastolic velocity from the LAD to the septal perforator from 41 +/- 9 to 72 +/- 17 cm/s (p < 0.0001). Three patients with HCM had surgical septal myectomy. These patients had luminal narrowings of the small intramural arteries at histopathologic examination. In conclusion, pulsed Doppler measurement of septal perforator flow velocities is feasible. In HCM, the epicardial coronary arteries enlarge to accommodate increased flow, and diastolic velocity is normalized. In contrast, the increased velocities in the septal branches of patients with HCM are similar to those previously observed in tunnel-like obstructions. These findings suggest that in HCM, notwithstanding an increase in coronary flow, hemodynamically significant narrowings are present in the septal branches. Doppler TTE may become useful for evaluation of abnormal intramural coronary flow in HCM. PMID- 16377294 TI - Comparison of real-time three-dimensional echocardiography to magnetic resonance imaging for assessment of left ventricular mass. AB - This is the first study to assess the feasibility and accuracy of real-time 3 dimensional echocardiography (RT-3DE) for the measurements of left ventricular (LV) mass in patients with congenital heart disease (CHD) compared with magnetic resonance imaging (MRI). Twenty patients (60% men) with CHD were evaluated by MRI and RT-3DE on the same day. Their mean age was 29 +/- 8 years (range 19 to 49). RT-3DE was performed with a Philips Sonos 7500 echocardiographic system and LV mass analyses with the assistance of TomTec software. The results for LV mass obtained by manual tracing were compared with Signa 1.5-T MRI data. The acquisition of RT-3DE data sets was feasible in all 20 patients. Nine patients (45%) had good, 5 patients (25%) moderate, and 6 patients (30%) poor image quality of the 3-dimensional data set. The time of 3-dimensional data acquisition was 4 +/- 2 minutes. Off-line image processing and tracing required approximately 11 +/- 3 minutes. A very good correlation was observed between RT-3DE data with sufficient image quality and MRI (r = 0.98, y = 0.96x + 4.1, SEE 9.8 g), with a mean difference of 2.0 +/- 20 g. Interobserver agreement was excellent (r = 0.99, y = 0.97x + 3.81), with a mean difference of -1 +/- 11 g. In conclusion, the assessment of LV mass from RT-3DE data is feasible in patients with CHD. The mass of an abnormally shaped left ventricle can be determined with high accuracy and low interobserver variability in patients with good or moderate echocardiographic image quality. PMID- 16377295 TI - Clinician agreement with perioperative cardiovascular evaluation guidelines and clinical outcomes. AB - The American College of Cardiology/American Heart Association (ACC/AHA) published guidelines for preoperative cardiac risk stratification in 1996. Although clinician practice may differ from the guidelines, it remains unclear whether deviation from these guidelines affects clinical outcomes. This study sought to determine if discordance between clinician practice and the ACC/AHA guidelines affects perioperative cardiac outcomes. Eight hundred twenty-three patients who underwent 864 consecutive preoperative evaluations performed from 1995 to 1997 at a tertiary care academic medical center were prospectively followed. Clinician recommendations for preoperative cardiac testing were compared with ACC/AHA guideline recommendations. Frequencies of perioperative cardiac complications were compared between concordant and discordant testing recommendations. There were 33 perioperative cardiac complications (3.8%). Overall, there was no difference in the frequency of complications when there was discordance with the ACC/AHA guidelines compared with concordance (4.1% vs 3.7%, p = 0.81). The ACC/AHA guidelines recommended cardiac testing for 236 patients (27.3%). Clinicians ordered testing in half of those cases (n = 112). There was a lower frequency of cardiac complications when clinicians did not perform testing as recommended by the ACC/AHA guidelines (3.2% vs 10.7%, p = 0.02). Conversely, clinicians ordered cardiac testing in 45 patients (7%) when not recommended by the guidelines. Patients in this group had a trend toward more cardiac complications (6.7% vs 2.4%, p = 0.09). In conclusion, the failure of clinicians to follow the ACC/AHA guidelines when perioperative testing was recommended did not result in a higher frequency of cardiac complications. PMID- 16377296 TI - Cardiopulmonary exercise testing and six-minute walk correlations in pulmonary arterial hypertension. AB - In a clinical trial of 178 patients with pulmonary arterial hypertension, treatment improved the 6-minute walk (6MW) distance but not the peak VO2. To clarify this discrepancy, we examined the exercise data from all study sites. Patients received either the endothelin receptor antagonist sitaxsentan or placebo and underwent serial 6MW and cardiopulmonary exercise testing (CPET). In 518 pairs of body weight-adjusted and unadjusted 6MW and CPET data, the correlation between 6MW and peak VO2 was 0.48. In the 4 sites with the highest overall correlation at baseline (r = 0.62 compared with 0.46 for the other 19 sites, p = 0.04), the correlations at baseline (0.66) and at week 12 (0.65) were similar (p = 0.90). However, the correlation increased significantly from baseline (0.34) to week 12 (0.54, p = 0.0005) for the other 19 sites. The correlations between weight-adjusted 6MW distance and peak VO2 for all centers (0.76) were significantly higher than the unadjusted correlations (0.48, p < 0.0001). Improvement in the correlations over time between the 6MW and CPET data at less-experienced sites was most consistent with improved technical skill with increasing experience. Weight adjustment of the 6MW improved its correlation with peak VO2. In conclusion, in future multicenter trials, CPET expertise should be validated at all sites before subject enrollment. PMID- 16377297 TI - Predictors of major hemorrhage following fibrinolysis for acute pulmonary embolism. AB - One hundred four patients at Brigham and Women's Hospital who received alteplase for acute pulmonary embolism were evaluated. Major bleeding occurred in 20 patients (19.2%). The principal site of bleeding was unknown in 9 (45.0%), gastrointestinal in 6 (30.0%), retroperitoneal in 3 (15.0%), intracranial in 1 (5.0%), and splenic in 1 (5.0%). Independent predictors of major hemorrhage were the administration of catecholamines for systemic arterial hypotension (odds ratio [OR] 115, 95% confidence interval [CI] 9.4 to 1,410.9, p < 0.001), cancer (OR 16.0, 95% CI 3.2 to 80, p = 0.004), diabetes mellitus (OR 9.6, 95% CI 1.7 to 54, p = 0.010), and elevated international normalized ratio before fibrinolysis (OR 5.8, 95% CI 1.5 to 22, p = 0.012). PMID- 16377298 TI - Consequences of increasing heart rate on deceleration time, the velocity-time integral, and E/A. AB - The ascendancy of diastolic heart failure to "epidemic" proportions has increased the use of and reliance on Doppler echocardiography as a source for diagnosis and as the preferred method for determining indexes of diastolic function (DF). Current indexes are primarily derived from shape-based features of Doppler E and A waves, such as their amplitudes, slopes, durations, and areas. Load dependence and pathologic correlates of these indexes have been considered, but DF indexes are not routinely corrected for heart rate (HR). To determine the dependence of selected Doppler-derived indexes of DF on HR, transmitral Doppler flow velocities and electrocardiograms were simultaneously recorded during supine bicycle exercise in 21 young, healthy volunteers. Standard E- and A-wave shape-based indexes (acceleration time, deceleration time [DT], peak E, peak A) were measured using triangle approximation. Velocity-time integrals (VTIs) were calculated by trapezoidal and triangular approximations. A-wave peak velocity (A) was measured conventionally, relative to baseline, and also using 2 alternative methods: A*, measured relative to the E@A velocity, and Ac, relative to the E-wave deceleration value at peak A-wave velocity. E/A was calculated conventionally and by using A* and Ac. The results showed that DF indexes derived from individual E waves are essentially HR independent. DT showed a mere 20% decrease for a 100% increase in HR. A triangular approximation for the E-wave VTI and the corrected E/Ac were found to be nearly HR independent. In conclusion, on the basis of the established continuity of cardiac output as a function of increasing HR and the observed data, Doppler-derived indexes of DF (DT, VTIs, E/Ac) can be treated as essentially HR independent only if the VTI and A-wave peak are corrected for HR as described. PMID- 16377299 TI - Quantitative estimation of left ventricular ejection fraction from mitral valve E point to septal separation and comparison to magnetic resonance imaging. AB - This study tested the hypothesis that the mitral valve E point-to-septal separation (EPSS) can be used to quantify the left ventricular (LV) ejection fraction (EF) on a continuous scale rather than simply as "normal" or "reduced." After excluding 5 patients with mitral valve prostheses, asymmetric septal hypertrophy, or significant aortic insufficiency, EPSS was measured in 42 patients by 3 independent observers on a cardiac magnetic resonance image identical to the echocardiographic parasternal long-axis view. In each patient, the reference standard LVEF was calculated from the magnetic resonance short-axis cross-sectional stack images by Simpson's rule and ranged from 11% to 72%. For all 42 patients, linear regression revealed the relation magnetic resonance imaging (MRI) LVEF = 75.5 - 2.5. EPSS (millimeters). Correlation between EPSS and the MRI LVEF for the 3 observers agreed closely, ranging from r = 0.78 to r = 0.82 (SEE 9 to 10), with similar regression coefficients. After blinded segmental wall motion scoring of the gated magnetic resonance cine images of the left ventricle in each patient, correlations, SEEs, and regression coefficients were found to be very similar in the 21 patients with the most homogenous wall motion, compared with the 21 patients with the most heterogenous wall motion. In conclusion, clinically useful quantitative prediction of the LVEF as a continuous variable can be obtained from the EPSS with a simple linear regression equation in a substantial portion of patients and may be a useful adjunct for assessment of LV function. PMID- 16377300 TI - Comparison of cardioprotective benefits of vigorous versus moderate intensity aerobic exercise. AB - Aerobic fitness, not merely physical activity, is associated with a reduced risk of cardiovascular disease. Vigorous intensity exercise has been shown to increase aerobic fitness more effectively than moderate intensity exercise, suggesting that the former may confer greater cardioprotective benefits. An electronic search of published studies using PubMed was conducted for 2 types of investigations, epidemiologic studies that evaluated the benefits of physical activity of varying intensity levels and clinical trials that trained individuals at different intensities of exercise while controlling for the total energy expenditure. A secondary search was conducted using the references from these studies. The epidemiologic studies consistently found a greater reduction in risk of cardiovascular disease with vigorous (typically > or =6 METs) than with moderate intensity physical activity and reported more favorable risk profiles for individuals engaged in vigorous, as opposed to moderate, intensity physical activity. Clinical trials generally reported greater improvements after vigorous (typically > or =60% aerobic capacity) compared with moderate intensity exercise for diastolic blood pressure, glucose control, and aerobic capacity, but reported no intensity effect on improvements in systolic blood pressure, lipid profile, or body fat loss. In conclusion, if the total energy expenditure of exercise is held constant, exercise performed at a vigorous intensity appears to convey greater cardioprotective benefits than exercise of a moderate intensity. PMID- 16377301 TI - Familial origin of atrial septal aneurysm. AB - The familial origin of atrial septal defects has been previously reported. This is the first study describing 2 families with atrial septal aneurysm of familial origin. The present study represents both clinically manifested and silent atrial septal aneurysms. Moreover, female gender predominance is also reported. Based on the presented data it could be suggested that all first-degree relatives of affected patients should be screened by cross-sectional echocardiography, particularly if they are women. In these patients, the use of aspirin might be the first line of treatment. PMID- 16377302 TI - When time is about to run out. PMID- 16377303 TI - Drawbacks of nonrandomized trials in acute coronary syndromes. PMID- 16377304 TI - "Improvement" in the placebo group could be due to regression to the mean as well as to sociobiologic factors. PMID- 16377305 TI - Usefulness of statins in protecting against ventricular arrhythmias. PMID- 16377306 TI - AMPK activation regulates apoptosis, adipogenesis, and lipolysis by eIF2alpha in adipocytes. AB - AMP-activated protein kinase (AMPK) is a metabolic master switch regulating glucose and lipid metabolism. Recently, AMPK has been implicated in the control of adipose tissue content. Yet, the nature of this action is controversial. We examined the effect on F442a adipocytes of the AMPK activator-AICAR. Activation of AMPK induced dose-dependent apoptotic cell death, inhibition of lipolysis, and downregulatation key adipogenic genes, such as peroxisome proliferator-activated receptor (PPARgamma) and CCAAT/enhancer-binding protein alpha (C/EBPalpha). We have identified the alpha-subunit of the eukaryotic initiation factor-2 (eIF2alpha) as a target gene which is phosphorylated following AICAR treatment. Such phosphorylation is one of the best-characterized mechanisms for downregulating protein synthesis. 2-Aminopurine (2-AP), an inhibitor of eIF2alpha kinases, could overcome the apoptotic effect of AICAR, abolishing the reduction of PPARgamma and C/EBPalpha and the lipolytic properties of AMPK. Thus, AMPK may diminish adiposity via reduction of fat cell number through eIF2alpha-dependent translation shutdown. PMID- 16377308 TI - Eosinophilic esophagitis in adults: clinical, endoscopic, histologic findings, and response to treatment with fluticasone propionate. AB - BACKGROUND: Eosinophilic esophagitis is an increasingly recognized disorder characterized by intense eosinophilic infiltration of the esophageal mucosa. The aim of this study was to define the clinical syndrome, the endoscopic features, and the distribution of the eosinophil infiltrate in adults with eosinophilic esophagitis. We undertook a prospective evaluation of the symptomatic and histologic response to treatment with fluticasone propionate. METHODS: Twenty-six patients (18 men; mean age 36 years) had symptom assessment and barium studies, esophageal motility recordings, and 24-hour esophageal pH studies. Upper-GI endoscopy was performed with quantitative eosinophil counts of biopsy specimens from the proximal and distal esophagus, the gastric antrum, and the duodenum. Nineteen subjects received 4 weeks of swallowed fluticasone propionate. After treatment, symptom assessment and endoscopic biopsies were repeated. RESULTS: All 26 patients had a history of dysphagia, and 11 presented acutely with food-bolus obstruction. Esophageal peristalsis was normal in most and gastroesophageal reflux coexisted in 10 patients. Characteristic endoscopic findings of furrows (20) and rings (18) were observed. All 19 treated patients had symptom improvement and a significant decrease in esophageal eosinophil counts. CONCLUSIONS: Eosinophilic esophagitis is a distinct entity that may coexist with gastroesophageal reflux. Swallowed fluticasone propionate is an effective treatment. PMID- 16377309 TI - Eosinophilic esophagitis: the endoscopist's enigma. PMID- 16377310 TI - Histologic results of EMR for esophageal lesions diagnosed as high-grade intraepithelial squamous neoplasia by endoscopic biopsy. AB - BACKGROUND: Biopsy specimens obtained from esophageal lesions detected in endoscopic screening with iodine staining have often been diagnosed as high-grade intraepithelial squamous neoplasia (WHO 2000). However, a management strategy for such lesions has not been established. The purpose of this study was to perform EMR for such lesions and to determine the actual tumor stage in patients with complete resection and the outcomes after EMR. PATIENTS: During the study period, 51 patients were found to have esophageal lesions diagnosed as high-grade intraepithelial squamous neoplasia by using endoscopic iodine staining in biopsy specimens. All of the patients underwent EMR, and resected specimens were reviewed microscopically. RESULTS: Histologic examination of totally resected specimens revealed that 12 (23.5%) of the 51 patients had tumor invasion of the lamina propria mucosae and that 4 (7.8%) had tumor invasion of the muscularis mucosae. The remaining 35 patients (68.6%) were confirmed to have high-grade intraepithelial squamous neoplasia. The invasive focus in all of the 16 lesions of invasive squamous-cell carcinoma was surrounded by high-grade intraepithelial squamous neoplasia. After a median of 23 months of follow-up, there were two recurrences, and those patients required second EMR. CONCLUSIONS: Histologic results suggested that high-grade intraepithelial squamous neoplasia of the esophagus has characteristics of carcinoma in the preinvasive stage. EMR should be performed for esophageal lesions diagnosed by endoscopic biopsy as high-grade intraepithelial squamous neoplasia, not only because of its probable malignant potential but also because more than 30% of such lesions are actually invasive carcinoma. PMID- 16377311 TI - Endoscopic measurement of cardia circumference as an indicator of GERD. AB - BACKGROUND: It is theorized that repeated gastric distention leads to dilatation of the cardia and the development of GERD. We hypothesize that cardia circumference correlates with the presence and the severity of GERD, and we developed software to measure cardia circumference from static endoscopic images. Our aims were to validate the software and to quantify cardia circumference along the spectrum of GERD. METHODS: Software-based measurements were compared with actual measurements in animal and mechanical models. A retrospective review of an endoscopic database and patient charts produced 273 subjects, grouped as follows: controls, GERD, < or = 3-cm Barrett's esophagus, or > 3-cm Barrett's esophagus. A blinded abstractor measured cardia circumference by using images from the database. RESULTS: Software and actual measurements correlated closely and were reproducible among observers. Median cardia circumference for each group was the following: control, 31.8 mm; GERD, 37.8 mm; < or = 3-cm Barrett's esophagus, 38.4 mm; and > 3-cm Barrett's esophagus, 45.0 mm (p < 0.001). By using 34.3 mm as a cutoff, cardia circumference was 85.3% sensitive and 89.6% specific for the diagnosis of GERD. CONCLUSIONS: There was a direct relationship between cardia circumference and the presence of GERD. This finding augments our understanding of the anatomic contributions of the esophagogastric junction in the pathogenesis of GERD. Cardia measurement may prove to be a useful diagnostic tool. PMID- 16377312 TI - Can endoscopy tell us anything about GERD in the absence of esophagitis or Barrett's esophagus? PMID- 16377313 TI - EUS characteristics of Nissen fundoplication: normal appearance and mechanisms of failure. AB - BACKGROUND: In patients who develop symptoms after Nissen fundoplication, the precise mechanism of failure can be difficult to determine. Current testing modalities do not demonstrate sufficient anatomic detail to definitively determine the mechanism. This observational study establishes that EUS can determine fundoplication integrity and hiatal anatomic relationships after Nissen fundoplication. METHODS: EUS was performed on the native esophagogastric junction and after Nissen fundoplication in two swine. The EUS characteristics of a properly performed fundoplication were determined. Subsequently, complications of Nissen fundoplication were created, and EUS was performed on each. The EUS criteria of each mechanism of failure were defined. RESULTS: EUS provided sufficient axial resolution to distinguish the esophagus, the fundoplication, and the surrounding hiatal structures within a single image. US of the native esophagogastric junction discerned the length of intra-abdominal esophagus, esophagogastric junction, crura, and anterior hiatus, and, thus, the point of entry into the abdominal cavity. EUS of Nissen fundoplication revealed a 5 layered pattern in a 360 degree configuration. These layers represent the following: (1) the esophageal wall, (2) the space between the esophagus and the fundoplication, (3) the inner gastric wall of the fundoplication, (4) the gastric lumen, and (5) the outer gastric wall of the fundoplication. A slipped repair was identified by the presence of an echogenic gastric serosa within the fundoplication. A tight fundoplication results in attenuation of the gastric walls, thickening of the esophageal wall, and loss of the 5-layer pattern secondary to obliteration of the potential spaces of the gastric lumen. Dehiscence of the fundoplication was evidenced by a less than 360 degree 5-layer pattern. CONCLUSIONS: EUS of hiatal anatomic relationships is feasible and provides detailed information regarding the integrity and the position of a Nissen fundoplication. EUS may enable a precise determination of the anatomic causes of failure after antireflux surgery. PMID- 16377314 TI - Looking for trouble: EUS after failed fundoplication. PMID- 16377315 TI - Argon plasma coagulation for early gastric cancer: technique and outcome. AB - BACKGROUND: Argon plasma coagulation (APC) is a noncontact technique for tissue coagulation. APC has been used to treat early gastric cancer in patients who cannot undergo EMR or open surgery, but a standard procedure for APC is lacking. OBJECTIVE: Our objectives were to assess the clinical usefulness of APC in patients with early gastric cancer. DESIGN: This was a small, retrospective pilot study. SETTING: All patients were treated at the Department of Gastroenterology, Kitasato University East Hospital, Sagamihara, Japan. METHODS: We studied 40 patients with early gastric cancer in whom both EMR and open surgery were contraindicated. The macroscopic tumor type was superficial elevated in 11 patients, superficial depressed in 27, and superficial elevated plus superficial depressed in two. The histologic classification was intestinal type in 37 patients and diffuse type in 3. INTERVENTION: From January 1998 through March 1999, all patients received one session of APC. From April 1999 through August 2001, all patients received two sessions of APC. From September 2001 through March 2002, an additional session of APC was given only to patients who had large protruding lesions, depressed lesions 2 cm or greater in diameter, or submucosal invasion. MAIN OUTCOME MEASUREMENTS: The main outcome measurements were residual tumor or recurrence of early gastric cancer. RESULTS: Intestinal-type intramucosal carcinoma disappeared after one or two sessions of APC. Submucosal and diffuse-type tumors had a high risk of residual tumor cells because of inadequate treatment after one session of APC. However, such lesions were locally controlled by follow-up APC. LIMITATIONS: This was a small, retrospective pilot study. Confirmation of long-term outcome is required. CONCLUSIONS: Small early gastric carcinomas can be successfully treated by a single session of APC. Larger protruding-type lesions and submucosal tumors are likely to require two sessions of APC. PMID- 16377316 TI - The art and science of painting in early gastric cancer: is there a role for ablation therapy? PMID- 16377317 TI - Pancreatic endoprostheses in chronic pancreatitis: criteria to predict stent occlusion. AB - BACKGROUND: Good clinical results of main pancreatic duct (MPD) stent placement in chronic pancreatitis (CP) are clouded by early stent occlusion. The aim of this study was to increase knowledge about stent occlusion and its effects on clinical symptoms, and to define criteria that enable the prediction of clogging. METHODS: A total of 100 pancreatic endoprostheses of 47 patients (32 men, 15 women; mean age, 53 years; standard deviation, 9 years) with CP were bench tested by simulating the pathophysiologically increased MPD pressure. The main study parameter was the reduction of water flow through clogged stents in comparison with native endoprostheses of identical type, length, and diameter. Major stent occlusion was defined as flow reduction by > or = 75%. The association between time to stent occlusion and stent- or patient-related variables was evaluated. RESULTS: Occlusion took place in nearly all endoprostheses (97%). No significant association of occlusion with clinical or blood parameters was found. Multifactorial analysis proved 4 risk factors for major stent occlusion: (A) stent diameter > 8.5F, (B) stent length > 8 cm, (C) female gender, (D) exocrine pancreatic insufficiency that required regular oral enzyme supplementation. According to the relative risk, these factors were given the following scores: A, 3 points; B to D, 2 points. Stents in patients with a score sum > 5 showed a significantly higher risk of major stent occlusion within 90 days. CONCLUSIONS: Stent clogging in CP seems to be an inevitable phenomenon. Because clinical and laboratory data do not reliably indicate clogging, stent removal or exchange should be performed in high-risk patients (score sum > 5) within 3 months. PMID- 16377318 TI - Pancreatic stents in chronic pancreatitis: do they function as a tube, a wick, or a placebo? PMID- 16377319 TI - Effectiveness of a new long cytology brush in the evaluation of malignant biliary obstruction: a prospective study. AB - BACKGROUND: Cancer detection rates with biliary brush sampling remain disappointingly low. A low cellular yield is often the limiting factor in making a diagnosis of malignancy. The new Cytolong brush (Cook Endoscopy, Winston-Salem, NC) is 3 mm in diameter, 5 cm long, with stiffer bristles oriented at 45 degrees on a 7F sheath. We hypothesized that this new brush might improve cancer detection rates by increasing cellular yield. METHODS: Patients found to have a biliary stricture suspicious for neoplasia on ERCP were randomized to undergo brush sampling for cytology with a standard Geenen brush (GB; Cook Endoscopy, Winston-Salem, NC) [3 mm in diameter, 1.5 cm long, bristles oriented at 90 degrees on a 6F sheath] or the Cytolong brush (CB). Repeat sampling was then performed with the other brush. Stricture dilation was not performed prior to brushing. Specimen results were considered normal, atypical (considered benign), highly atypical (suspicious for cancer), or malignant. All specimens were assigned a cellularity score (0 to 3, insufficient to excellent). Final diagnosis was based on cytologic results plus surgery, EUS, autopsy, or clinical follow-up. RESULTS: From November 2001 to July 2003, 102 patients had specimens obtained from 94 malignancies (47% pancreatic cancer). The cancer detection rate was 25 of 94 (27%) using CB and 28 of 94 (30%) with GB (p = NS). No patient had positive cytology results with CB and negative cytology results with GB. The yield of the two brushes combined was 28 of 94 (30%). Cancer detection rates of 28% (18 of 64) and 31% (20 of 64) were found for CB and GB, respectively, in distal biliary strictures, and 23% (7 of 30) and 27% (8 of 30) in proximal strictures (p = NS). Insufficient or limited cellularity was seen less frequently with CB (11 of 98) than with GB (17 of 98), and the mean cellular yield was greater with CB than GB (2.6 vs 2.4, p = 0.006). SUMMARY: Despite improved cellularity, cancer detection rates were not improved by using the larger Cytolong brush in this study. There was no statistical difference between the brushes in both proximal and distal biliary strictures. CONCLUSIONS: The yield of biliary brush cytology at ERCP remains low. Increasing brush size and bristle stiffness does not increase detection rates. Newer devices and processing techniques are required to allow detection rates to approach those attained in other GI tract malignancies. PMID- 16377320 TI - Brush cytology and the diagnosis of pancreaticobiliary malignancy during ERCP. PMID- 16377321 TI - Double-balloon enteroscopy in the diagnosis and the management of small-bowel diseases: an initial experience in 40 patients. AB - BACKGROUND: Double-balloon enteroscopy (DBE) is a new technique, recently described by its innovator, and it is seen to be beneficial in the diagnosis and management of small-bowel disease. OBJECTIVE: To evaluate this new technique in consecutive patients with suspected small-bowel disease. DESIGN: Single-center prospective study. SETTING: Tertiary referral hospital, conducted from July to December 2004. PATIENTS: Forty consecutive patients with suspected small-bowel disease referred for DBE, mean age of 58 years (range, 14-89 years), 17 men. INTERVENTIONS: Endoscopic biopsies or therapy was performed as clinically indicated. MAIN OUTCOME MEASUREMENTS: Significant diagnostic input and therapeutic intervention based on clinical outcomes. RESULTS: Indications included obscure GI bleeding (18), iron deficiency anemia (6), anemia of chronic disease (4), acute obscure GI bleeding (4), abdominal pain with other symptoms (4), Crohn's disease (3), and abdominal pain alone (1). Nineteen patients (47.5%) had a small-bowel finding, with 30 of 40 (75%) of the patients having a significant diagnostic input. Intervention was performed in 13 (32.5%) patients with success in 10 (77%). The only variable significantly associated with therapeutic success was a previous history of blood transfusions (p < 0.01). This was the only independent predictor identified by multiple logistic regression analysis (Odds ratio 13.5: 95% confidence interval [1.5, 120]). One perforation from contact diathermy occurred. In 10 attempts at total enteroscopy, none were successful. LIMITATIONS: Nonblinded nonrandomized study. CONCLUSIONS: These early data suggest DBE to be effective in the diagnosis and the therapy of small-bowel disease, particularly those patients with a history of blood transfusion. PMID- 16377322 TI - Diphenhydramine as an adjunct to sedation for colonoscopy: a double-blind randomized, placebo-controlled study. AB - BACKGROUND: Intravenous benzodiazepines in combination with opiates are used to achieve moderate sedation for colonoscopy. Although effective, these agents have potential adverse effects, such as respiratory depression and hypotension. Diphenhydramine hydrochloride possesses central nervous system depressant effects that theoretically could provide a synergistic effect for sedating patients. OBJECTIVE: The objective was to assess the efficacy of adding diphenhydramine hydrochloride as an adjunct to improve sedation and to reduce the amount of standard sedatives used during colonoscopy. DESIGN: We conducted a prospective, randomized, double-blind, placebo-controlled study. SETTING: The study was conducted in a university hospital with an active GI fellowship training program. PATIENTS: The study group comprised 270 patients undergoing screening/diagnostic/therapeutic colonoscopy were enrolled. INTERVENTIONS: Patients were randomized to receive either 50 mg of diphenhydramine or placebo, given intravenously 3 minutes before starting conscious sedation with intravenous midazolam and meperidine. MAIN OUTCOME MEASUREMENTS: The main outcome measure was anesthetic effect as assessed by the endoscopy team and by the patient; quantity of adjunctive sedatives to achieve adequate sedation. RESULTS: Of 270 patients, data were analyzed for 258 patients, with 130 patients in the diphenhydramine group and 128 patients in the placebo group. There was a 10.1% reduction in meperidine usage and 13.7% reduction in midazolam usage in favor of the diphenhydramine group. The mean evaluation scores as judged by the faculty, the fellows, and the nurses were statistically significant in favor of the diphenhydramine group. In addition, patient scores for overall sedation and pain level favored the group that received diphenhydramine. CONCLUSIONS: Intravenous diphenhydramine given before initiation of standard sedation offers a significant benefit to conscious sedation for patients undergoing colonoscopy. PMID- 16377323 TI - Sedation and analgesia for GI endoscopy. PMID- 16377324 TI - Improving compliance with postpolypectomy surveillance guidelines: an interventional study using a continuous quality improvement initiative. AB - BACKGROUND: Despite guidelines, physicians tend to perform postpolypectomy surveillance colonoscopies too frequently. OBJECTIVE: The objective of the study was to determine the baseline compliance rate with postpolypectomy guidelines in our unit and to determine the influence of a continuous quality improvement (CQI) intervention on improving the compliance rate and on decreasing the potential additional costs because of the scheduling of postpolypectomy surveillance colonoscopies earlier than indicated. DESIGN: This was a single-arm, pretest posttest design. SETTING: This study took place at a tertiary care, academic medical center. PATIENTS: The medical records of all patients who underwent colonoscopy with polypectomy in our unit retrospectively during 6 months preceding (baseline period) and prospectively for 6 months after an intervention (postintervention period) were reviewed for patient demographics, colonoscopy findings, and scheduling of repeat colonoscopies. INTERVENTION: We used 3 components: (1) distribution of a wallet-size card with a summary of postpolypectomy guidelines to all endoscopists, (2) placement of guideline charts near computers used for typing endoscopy reports, and (3) distribution and reinforcement of the guidelines in a monthly continuous quality improvement meeting. MAIN OUTCOME MEASURES: The main outcome measures were compliance rates, mean times to repeat colonoscopy, and additional costs from surveillance colonoscopies being scheduled earlier than indicated were compared between the two periods. RESULTS: There were 278 patients in the baseline period and 242 in the postintervention period, with similar patient and polyp characteristics. After the intervention, the compliance rate with guidelines improved from 57.2% to 81% (p < 0.001). The mean time to a repeat colonoscopy increased from 4.5 to 5.2 years (p = 0.003) (i.e., a 14% reduction in the number of postpolypectomy surveillance colonoscopies performed per year). This would result in a reduction of a total of 73 surveillance colonoscopies per year in our unit, with a projected cost savings of 171,331 dollars per year (cost of a colonoscopy assumed at 2347 dollars). LIMITATIONS: The limitation of the study was possible enhanced performance secondary to being observed (Hawthorne effect). Because more than 1 intervention was used, we do not know which one is more effective. CONCLUSIONS: Relatively simple and easy-to-implement quality improvement initiatives can significantly enhance compliance with postpolypectomy guidelines and result in cost savings because of a reduction in the number of postpolypectomy surveillance colonoscopies being scheduled earlier than recommended guidelines. PMID- 16377325 TI - Reducing unnecessary surveillance colonoscopies: a mandate for endoscopists. PMID- 16377327 TI - The importance of having a mentor, and how to find one. PMID- 16377326 TI - International survey of knowledge of indications for EUS. AB - BACKGROUND: The knowledge level for EUS indications among gastroenterologists across different locations and practices is not known. The aim of this study was to assess knowledge of EUS indications among a diverse group of gastroenterologists, both nationally and internationally. METHODS: A web-based survey was designed to assess knowledge of EUS with respect to 4 organ systems: esophagus, gastroduodenum, hepatopancreatobiliary, and colorectum. The survey was distributed by electronic mail (e-mail) to members of the American Society for Gastrointestinal Endoscopy. RESULTS: The survey was distributed to 3848 physicians, of whom 2848 had an active e-mail address. There were 323 respondents (11.3%), of whom 210 were U.S. members and 113 international. Overall, the mean score for the different organ systems for all respondents was highest for gastroduodenum (93% correct) and lowest for colorectum (71%) compared with esophagus (79%) and hepatopancreatobiliary (83%) systems. The mean total score was higher for U.S. respondents (84% correct) compared with international respondents (79%, p < 0.0001). Endosonographers fared better than those who were not endosonographers (85% vs. 81%, p = 0.0002). CONCLUSIONS: Knowledge levels of colorectal applications of EUS are poorest among the 4 organ systems studied. Future educational initiatives should focus on applications of EUS in this category. Studies are required to assess the impact of this education on the appropriateness of EUS referral patterns. PMID- 16377328 TI - Colonoscopy in the sitting position: lessons learned from self-colonoscopy by using a small-caliber, variable-stiffness colonoscope. PMID- 16377329 TI - CT colonography for colon cancer screening. PMID- 16377330 TI - A new esophageal stent design (Niti-S stent) for the prevention of migration: a prospective study in 42 patients. AB - BACKGROUND: Covered, rather than uncovered, metal stents are used for the palliation of dysphagia from esophageal cancer, but a major drawback is the risk of stent migration, which occurs in up to 20% of patients. To overcome this problem, a double-layered stent, the Niti-S stent (Taewong Medical, Seoul, Korea), has been developed. The Niti-S stent consists of an inner polyurethane layer to prevent tumor ingrowth and an outer uncovered nitinol wire tube to allow the mesh of the stent to embed itself in the esophageal wall. METHODS: Between June 2003 and May 2004, 42 patients with malignant dysphagia caused by inoperable carcinoma of the esophagus or gastric cardia were treated with a Niti-S stent. Patients were prospectively followed and data collection focused on recurrent dysphagia, functional outcome, complications, and survival. RESULTS: At 4 weeks, the dysphagia score had significantly improved from a median of 3 (liquids only) to 0 (ability to eat a normal diet). Five of 42 (12%) patients with a Niti-S stent developed recurrent dysphagia, mainly due to tissue overgrowth (2 of 42; 5%) and stent migration (3 of 42; 7%). Major complications (perforation, 1; aspiration pneumonia, 2; hemorrhage, 2) occurred in 5 of 42 (12%) patients. Pain following stent placement was observed in 5 of 42 (12%) patients, and symptomatic gastroesophageal reflux occurred in 2 of 42 (5%) patients. CONCLUSIONS: The Niti S stent provides symptomatic relief of malignant dysphagia and effectively reduces recurrent dysphagia. Its double-layered design is probably important in preventing migration. In addition, the complete covering of the Niti-S stent may be a factor in preventing tissue overgrowth at both ends of the stent. PMID- 16377331 TI - Colon cleaning during colonoscopy: a new mechanical cleaning device tested in a porcine model. AB - BACKGROUND: Poor colon cleaning frequently impairs colonoscopy. The aim of the study was to develop a method of cleaning the colon during colonoscopy. METHODS: A soft-tipped catheter with a water jet spray at its tip was designed to break up and propel stool contents in the direction of the anus. Stool contents were collected in a container attached to an endoscopy trolley. OBSERVATIONS: In bench tests, colon models filled with porridge were rapidly cleaned. Unprepared colons (n = 15) of anesthetized pigs were cleared in a few minutes by using this device at colonoscopy, by advancing the catheter under direct vision into concretions, which were rapidly broken down. There was minimal trauma to mucosa in these survival studies. CONCLUSIONS: A colon cleaning method for use at colonoscopy was highly effective in cleaning colon models and in unprepared pig colon. This device may have a role in cleaning the colon in unprepared or poorly prepared patients. PMID- 16377332 TI - Colonoscopic polypectomy in retroflexion. AB - BACKGROUND: Little has been written about the value of retroflexion in the removal of large sessile colon polyps. OBJECTIVE: The objective of the study was to evaluate the utility of retroflexion for removal of large sessile colon polyps. DESIGN: This was a retrospective evaluation of consecutive cases. SETTING: This study was conducted at an academic-hospital-based tertiary-referral colonoscopy practice. PATIENTS: The study comprised consecutive patients with sessile polyps > or = 2 cm who were undergoing endoscopic resection. INTERVENTIONS: The intervention was endoscopic resection of 59 consecutive sessile colon polyps 2 cm or larger in size and located proximal to the rectum by using prototype colonoscopes with short bending sections. MAIN OUTCOME MEASURES: The main outcome measurement was successful endoscopic resection. RESULTS: Fourteen of the polyps were removed either entirely (n = 4) or partially (n = 10) in retroflexion. Patients with polyps that were removed in retroflexion were more likely to have been referred by another colonoscopist than those patients with polyps removed entirely in the forward view (p = 0.05). There were no perforations and no complications related to retroflexion. LIMITATIONS: The study is retrospective, and the practice is a tertiary referral colonoscopy practice. The colonoscopes used are not widely available at this time. CONCLUSIONS: Retroflexion is a useful adjunctive procedure for the removal of some colon polyps proximal to the rectum that are difficult to access endoscopically. The use of retroflexion can increase the fraction of proximal sessile colon polyps amenable to endoscopic resection. PMID- 16377333 TI - Optimizing resection of difficult colon polyps. PMID- 16377334 TI - Foreign body "bezoar". PMID- 16377335 TI - Cowden disease. PMID- 16377336 TI - A "hidden treasure" causing microcytic hypochromic anemia. PMID- 16377337 TI - Peutz-Jeghers gastric polyposis. PMID- 16377338 TI - EUS-guided core biopsy for pseudopapillary tumor of the pancreas. PMID- 16377339 TI - Foreign body in the common bile duct for 15 years. PMID- 16377340 TI - Electrocautery therapy for refractory anastomotic strictures of the esophagus. AB - BACKGROUND: Anastomotic esophageal stenoses after esophageal resection are common and sometimes are refractory to Savary bougie dilation. The efficacy of electrocautery needle-knife treatment in these patients is described. METHODS: Twenty patients with a refractory anastomotic stricture of the esophagus were treated with electrocautery and were followed for 12 months. All patients had recurrence of dysphagia despite repeated bougienage. OBSERVATIONS: All 12 patients with a stricture shorter than 1 cm remained without dysphagia after a single treatment. In all 8 patients with a long-segment stenosis of 1.5 to 5 cm, dysphagia recurred, and a mean of 3 treatments were necessary. The interval between electrocautery treatments was significantly longer compared with bougienage. There were no complications. The body weight of all patients increased. CONCLUSIONS: Electrocautery seems to be a good single-treatment modality for refractory short-segment anastomotic strictures, whereas longer segment stenoses appear to require repeated treatment sessions before similar results are obtained. PMID- 16377341 TI - Benign refractory esophageal strictures: widening the endoscopist's role. PMID- 16377342 TI - Sole treatment of lichen planus-associated esophageal stenosis with injection of corticosteroids. PMID- 16377343 TI - Systemic silicosis that involves the pancreas. PMID- 16377344 TI - Epstein-Barr virus-associated cholangiopathy: a new disease entity? PMID- 16377345 TI - Richter's hernia after colonoscopy: a rare complication. PMID- 16377346 TI - Successful endoscopic en bloc resection of a large laterally spreading tumor in the rectosigmoid junction by endoscopic submucosal dissection. PMID- 16377347 TI - Esophageal perforation after placement of wireless Bravo pH probe. PMID- 16377348 TI - Heterotopic gastric mucosa in the upper esophagus manifesting as a polypoidal mass causing dysphagia. PMID- 16377349 TI - One-piece endoscopic resection of a gastric tube cancer on the surgical staple line. PMID- 16377350 TI - Dissection of a refractory pancreatic-duct stricture by using a pre-cutting needle-knife. PMID- 16377351 TI - Ulcerated small-intestine duplication cyst: an unusual source of GI bleeding revealed by wireless capsule endoscopy. PMID- 16377353 TI - Salvage photodynamic therapy by using porfimer sodium after chemoradiation therapy: a Western viewpoint. PMID- 16377354 TI - How should gastric fundal varices be treated after achieving hemostasis? PMID- 16377356 TI - Pancreaticobiliary Heimlich maneuver to facilitate stent placement during ERCP. PMID- 16377358 TI - Perinatal depression. PMID- 16377359 TI - Risk factors for early postpartum depressive symptoms. AB - OBJECTIVE: Postpartum depressive disorders are common and symptoms may appear as early as the first 2 weeks postpartum. Data regarding hormone-related risk factors for depressive symptoms occurring in the very early postpartum period are scarce and may be of importance in identifying serious postpartum illness. We examined the association between the reported history of psychiatric symptoms of possible hormonal etiology and very early postpartum depressive symptoms. METHODS: All women (n= 1,800) in a general hospital maternity ward were assessed during the first 3 days after parturition for potential risk factors for postpartum depressive disorders by a self-reported questionnaire and for present mood symptoms (Edinburgh Postnatal Depression Scale, EPDS). The associations between potential risk factors and postpartum depressive symptoms were analysed. RESULTS: The incidence of women with an EPDS >or=10 was 6.8% (88/1,286). Significant risk factors for early postpartum depressive symptoms were a history of mental illness including past postpartum depression (PPD), premenstrual dysphoric disorder (PMDD), and mood symptoms during the third trimester. CONCLUSION: In accordance with other studies, a history of depression was found to be a risk factor for early postpartum mood symptoms. An association was also found between some risk factors of possible hormone-related etiology such as PMDD and third trimester mood symptoms and early postpartum mood symptoms. As such, early postpartum symptoms may indicate vulnerability to subsequent PPD; it may be of importance to assess these risk factors and mood immediately after parturition. A prospective study is needed to determine which of these risk factors is associated with progression to PPD and which resolves as the blues. PMID- 16377360 TI - Where is the patient? The association of psychosocial factors and missed primary care appointments in patients with diabetes. AB - OBJECTIVE: Missed appointments are associated with poorer health outcomes. We predicted that compared to secure attachment style, fearful and dismissing attachment styles would be associated with greater number of missed primary care visits in patients with diabetes. METHODS: In patients with diabetes from nine health maintenance organization primary care clinics, we collected data on attachment style and major depression status, and determined the number of missed primary care appointments from automated data. We used Poisson and logistic regression analyses to determine if attachment style was associated with the number of missed primary care same day appointments, scheduled office visits and scheduled preventive care visits, after adjusting for demographics, clinical characteristics, appointment frequency and clustering by clinic. We included major depression as a potential effect modifier. RESULTS: Among 3,923 patients with diabetes, prevalence rates of attachment styles were 43.9% for secure, 35.8% for dismissing, 8.1% for preoccupied and 12.2% for fearful attachment style. Major depression was present in 12.4% of patients. Among patients without major depression, there were more missed scheduled office visits (RR=1.46, 95% CI=1.18 1.81) among those with dismissing compared to secure attachment style. The likelihood of having missed same day appointments was lower for those with fearful attachment style relative to those with secure attachment style in nondepressed patients compared to patients with fearful and secure attachment style with major depression (P < .01). CONCLUSIONS: Attachment styles characterized by low levels of collaboration are associated with more missed primary care appointments compared to secure attachment style in patients with diabetes. These associations are moderated by depression status. PMID- 16377361 TI - Design and implementation of the telemedicine-enhanced antidepressant management study. AB - OBJECTIVE: Evidence-based practices designed for large urban clinics are not necessarily transportable into small rural practices. Implementing collaborative care for depression in small rural primary care clinics presents unique challenges because it is typically not feasible to employ on-site mental health specialists. The purpose of the Telemedicine-Enhanced Antidepressant Management (TEAM) study was to evaluate a collaborative care model adapted for small rural clinics using telemedicine technologies. The purpose of this paper is to describe the TEAM study design. METHOD: The TEAM study was conducted in small rural Veterans Administration community-based outpatient clinics with interactive video equipment available for mental health, but no on-site psychiatrists/psychologists. The study attempted to enroll all patients whose depression could be appropriately treated in primary care. RESULTS: The clinical characteristics of the 395 study participants differed significantly from most previous trials of collaborative care. At baseline, 41% were already receiving primary care depression treatment. Study participants averaged 5.5 chronic physical health illnesses and 56.5% had a comorbid anxiety disorder. Over half (57.2%) reported that pain impaired their functioning extremely or quite a bit. CONCLUSIONS: Despite small patient populations in rural clinics, enough patients with depression can be successfully enrolled to evaluate telemedicine-based collaborative care. PMID- 16377362 TI - Treating minority patients with depression and anxiety: what does the evidence tell us? AB - OBJECTIVE: The purpose of this study is to examine the current state of knowledge regarding treating ethnic/racial minority patients with mood and anxiety disorders, emphasizing data-based studies whenever possible. METHOD: This article reviews the evidence on poorer access and quality of care for minorities, the biological and cultural differences between minority and majority populations that may impact care and outcomes, and recent studies that address minority treatment response and outcomes both alone and in comparison to majority groups. RESULTS: Numerous impediments to appropriately treating anxious and depressed minority patients remain. Underutilization and poor quality of mental health care in minorities is due to less-than-favorable illness and treatment beliefs that affect adherence and outcome, stigma, clinician failure to engage the patient, poor patient activation and biological differences that may impact pharmacotherapy choice. However, though limited in number, some studies do indicate that when appropriate treatment is well-delivered to minorities, results are comparable to those seen among Caucasian patients. CONCLUSIONS: The clinician treating members of minority groups must consider differential personal elements, from the biological to the cultural, to achieve treatment success. The limited available data do suggest that minority patients can be successfully treated with available interventions. Of primary importance is for researchers to increase the number of carefully designed intervention studies that allow for ethnic/racial minority-specific analyses. PMID- 16377363 TI - Correlates of alcohol use among anxious and depressed primary care patients. AB - OBJECTIVE: The purpose of this study is to determine the patterns of alcohol use for primary care patients with anxiety disorders and/or major depression in three urban university-affiliated outpatient clinics. METHOD: A waiting room sample of adults was screened for anxiety disorders and major depression. Six hundred fourteen screened patients were assessed using the Composite International Diagnostic Interview [World Health Organization. Composite International Diagnostic Interview (CIDI) 2.1. Geneva: World Health Organization, 1997] and frequency-quantity alcohol use questions. Adjusted for age and gender, logistic regression analyses were used to determine associations between panic disorder, social phobia, PTSD, major depression and typical heavy (three drinks/two or more times a week) and frequent (four or more times a week) alcohol use. RESULTS: Of the patients, 6.19% (38/614) reported typical heavy drinking and 8.31% (51/614) reported frequent drinking in the preceding 3 months. PTSD was associated with heavy drinking (adjusted OR = 3.1; 95% CI, 1.3-7.3). Panic disorder was associated with frequent alcohol use (adjusted OR = 2.2; 95% CI, 1.2-4.2) but reduced heavy drinking (adjusted OR = 0.4; 95% CI, 0.2-0.9). There was no significant relationship between alcohol use and the co-occurrence of two or more anxiety and/or mood disorders. CONCLUSION: In an examination of primary care patients diagnosed, the majority of whom were with at least one anxiety disorder and/or major depression; current heavy and frequent alcohol use was associated with specific individual anxiety disorders and/or major depression. PMID- 16377364 TI - Psychological distress symptoms in women undergoing medical vs. surgical termination of pregnancy. AB - OBJECTIVES: The purpose of this study is to compare the baseline psychological distress and symptom profile of women undergoing either medical (with mifepristone) or surgical pregnancy termination and the psychological outcome 2 weeks after the procedure. METHODS: Women (n = 200) given free choice of pregnancy termination method, either medical or surgical, were assessed before pregnancy termination by a demographic questionnaire including questions regarding the choice of the method of pregnancy termination, the Brief Symptom Inventory (BSI), the Spielberger State Anxiety questionnaire and the Rotter Locus of Control Scale. Two weeks after the procedure, the BSI and Spielberger questionnaires were repeated. RESULTS: Women with a smaller number of past pregnancies tended to choose the medical procedure. Reasons for choosing the medical procedure were fear of surgery, anesthesia and of future fertility difficulties. Prior to the procedure, the "medical group" had significantly higher levels of obsessive-compulsive symptoms, guilt and BSI general symptom index score, and a trend for higher interpersonal sensitivity and paranoid ideation. Postprocedure, both groups showed significant decline in anxiety levels and did not differ on most symptom parameters. CONCLUSIONS: Women who chose to have a medical termination are marginally more symptomatic before the procedure than women choosing surgical termination. However, both methods of pregnancy termination resulted in significant reduction in preabortion psychological distress level. PMID- 16377365 TI - Prevalence and detection of alcohol use disorders among general hospital inpatients in eastern Taiwan. AB - OBJECTIVE: To examine (1) the prevalence of alcohol use disorders (AUD) and its various correlates, and (2) the detection rate of AUD by nonpsychiatric physicians and its related factors associated with increased chances of detection among inpatients in a general hospital in rural eastern Taiwan. METHODS: A well trained psychiatrist interviewed all adult patients admitted during a 1-month period using the Alcohol Inventory and reviewed all medical records for alcohol related diagnoses. RESULTS: A total of 303 inpatients aged 18 to 93 years were evaluated, of whom 78 (25.7%) were diagnosed by the psychiatrist as having AUD within the past year. Males, aborigines, middle-aged, current smokers and betel quid chewers had a significantly higher odds ratio for AUD. Nonpsychiatric physicians detected only 14.1% patients with recent AUD. Internists identified AUD nearly four times as often as surgeons. Better detection was associated with higher level of alcohol intake. CONCLUSIONS: This study demonstrates the high prevalence of AUD among hospitalized patients in eastern Taiwan and the frequent failure of physicians to identify the disorder. These findings suggest that more effort should be directed toward increasing medical professionals' awareness of AUD in general hospital settings, especially among high-risk groups. PMID- 16377366 TI - Validity and reliability of the Hospital Anxiety and Depression Scale in a hypertrophic cardiomyopathy clinic: the HADS in a cardiomyopathy population. AB - OBJECTIVE: The purpose of this study is the validation of the Hospital Anxiety and Depression Scale (HADS) in patients suffering from hypertrophic cardiomyopathy in an inner city teaching hospital. The secondary objective was to establish whether the use of the total HADS score to detect "caseness" is justified. METHODS: One hundred fifteen patients in a cardiac outpatient clinic completed the HADS, which was compared against the gold standard Structured Clinical Interview for DSM-III-R nonpatient version (SCID-np). Receiver operating characteristic (ROC) curves were created for the anxiety and depression subscales, as well as the total score, then sensitivity, specificity, likelihood ratios and positive and negative predictive values were calculated. Retest correlation was assessed at 2 weeks in 31 patients. RESULTS: The depression subscale was highly sensitive (100%) when the cutoff score 8 was used; however, the corresponding specificity was poor (79%). Raising the cutoff to 10 improved specificity but compromised sensitivity. The anxiety subscale was performed similarly though with less accuracy. The total HADS score produced a poor ROC curve and performed best when the cutoff was 14. CONCLUSIONS: The HADS performs well as a screening instrument for anxiety and depression in this population at the designed cutoff score 8. However, its use as a research instrument and the practice of using the total score to detect caseness are not supported by this study. PMID- 16377367 TI - Response to 4-month treatment with reboxetine in Parkinson's disease patients with a major depressive episode. AB - OBJECTIVE: The aim of this study is to evaluate response to reboxetine in a 4 month follow-up study on depression in Parkinson's disease (PD), and to assess its tolerability profile. METHODS: A prospective 4-month follow-up study was performed in 17 PD patients with a major depressive episode. The intensity of depressive symptoms was evaluated mainly with the Hamilton Rating Scale for Depression (HAM-D), and PD was assessed with the Unified Parkinson Disease Rating Scale (UPDRS). RESULTS: Reboxetine causes a progressive decrease in depressive symptoms in PD patients; the initial score of 16.76 (2.68) on HAM-D decreased to 5.85 (2.42) at 4 months (P < .002). Mean UPDRS scores did not show a statistically significant increase: 18.18 (2.6) at the beginning and 18.25 (2.4) at the end of the follow-up period (P = .8). CONCLUSIONS: Reboxetine, as first choice treatment for major depressive episodes in PD patients, seems to be effective in progressively improving depressive symptoms over the first 4 months of treatment until complete remission. Reboxetine does not seem to increase PD symptoms, whereas patients' quality of life improves. PMID- 16377368 TI - Substance abuse and recovery in a Psychiatric Intensive Care Unit. AB - OBJECTIVES: The purpose of this study is to compare the development in symptoms, behaviors, function and treatment between patients with or without a substance use (SU) diagnose in a Psychiatric Intensive Care Unit (PICU). METHODS: A total of 118 admitted patients were assessed at admittance, day 3 and discharge from the PICU. Symptoms of psychopathology, therapeutic steps taken, violent episodes and length of patient stay were recorded. RESULTS: More males than females received an SU diagnosis. Substance use patients had less psychiatric symptoms at admittance and showed a faster symptom reduction, more favorable and faster improvement of function and a shorter length of stay. Except for symptom reduction and shorter length of stay, these differences were largely due to differences in sex and diagnoses in the two groups. CONCLUSION: In a naturalistic group of patients in a PICU, SU is associated with favorable outcomes compared to patients not using substances. PMID- 16377369 TI - Validity of the Brief Patient Health Questionnaire Mood Scale (PHQ-9) in the general population. AB - OBJECTIVE: The aim of this study was to assess the validity of the Patient Health Questionnaire depression module (PHQ-9). It has been subject to studies in medical settings, but its validity as a screening for depression in the general population is unknown. METHOD: A representative population sample (2,066 subjects, 14-93 years) filled in the PHQ-9 for diagnosis [major depressive disorder, other depressive disorder, depression screen-positive (DS+) and depression screen-negative (DS-)] and other measures for distress (GHQ-12), depression (Brief-BDI) and subjective health perception (EuroQOL; SF-36). RESULTS: A prevalence rate of 9.2% of a current PHQ depressive disorder (major depression 3.8%, subthreshold other depressive disorder 5.4%) was identified. The two depression groups had higher Brief-BDI and GHQ-12 scores, and reported lower health status (EuroQOL) and health-related quality of life (SF-36) than did the DS- group (P's < .001). Strong associations between PHQ-9 depression severity and convergent variables were found (with BDI r = .73, with GHQ-12 r = .59). CONCLUSION: The results support the construct validity of the PHQ depression scale, which seems to be a useful tool to recognize not only major depression but also subthreshold depressive disorder in the general population. PMID- 16377371 TI - Psychiatric symptoms in progressive supranuclear palsy. PMID- 16377370 TI - A case of serotonin syndrome precipitated by abuse of the anticough remedy dextromethorphan in a bipolar patient treated with fluoxetine and lithium. PMID- 16377372 TI - Electroconvulsive therapy for the treatment of multiple system atrophy with major depression. PMID- 16377373 TI - Ziprasidone-related agranulocytosis following olanzapine-induced neutropenia. PMID- 16377374 TI - Delusional parasitosis or Ekbom syndrome: a case series. PMID- 16377375 TI - Minimal change disease in association with fire coral (Millepora species) exposure. AB - Numerous agents have been associated with minimal change disease. We describe a previously unreported association in a 45-year-old white woman of scuba diving exposure to fire coral (Millepora species) that was followed by the development of nephrotic syndrome, acute renal failure, pulmonary edema, and intubation. The renal biopsy specimen was consistent with minimal change disease. Institution of corticosteroid therapy resulted in symptomatic improvement and resolution of proteinuria. Physicians, particularly those in scuba-diving areas, should consider minimal change disease in exposed patients with proteinuria because a prompt diagnostic and therapeutic approach may potentially limit complications. PMID- 16377376 TI - Focal segmental glomerular sclerosis in a patient with neurofibromatosis type I. AB - A 22 -year-old white man was found to have a serum creatinine level of 3.4 mg/dL (259 micromol/L) and 6 g of protein in a 24-hour urine collection. Laboratory studies performed 5 years prior showed no evidence of abnormal renal function. Neurofibromatosis had been diagnosed in infancy, and genetic testing showed that the patient had a mutation in chromosome 17, consistent with neurofibromatosis type I. Mutations on chromosomes 1, 11, and 19 have been reported to result in genetic forms of focal segmental glomerular sclerosis. This is the first case report of focal segmental glomerular sclerosis occurring in a patient with neurofibromatosis type I. PMID- 16377377 TI - Macroscopic hematuria in a kidney transplant recipient: a rare cause. AB - Pseudoaneurysm formation in a renal transplant is an uncommon complication of such interventional procedures as percutaneous nephrostomy or renal biopsy; symptoms may be delayed for days or even years. Presentation may vary from incidental discovery to worsening renal insufficiency to life-threatening hemorrhage. We report a case of macroscopic hematuria from a pseudoaneurysm that developed in a kidney transplant recipient after placement of a percutaneous nephrostomy tube. This patient was treated with transcatheter embolization, which is highly effective. A high index of suspicion, along with early diagnosis and transcatheter embolization, are essential for the management of hematuria caused by pseudoaneurysm formation from percutaneous nephrostomy tube placement. PMID- 16377378 TI - Levofloxacin penetration into a renal cyst in a patient with autosomal dominant polycystic kidney disease. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disorder often complicated by cyst infection. Treatment becomes particularly challenging when the culprit organism is resistant to antibiotics known to sufficiently penetrate cysts. We present the case of a patient with ADPKD and suspected cyst infection caused by group B streptococcus who was treated successfully with levofloxacin and ampicillin. Although data support excellent cyst penetration with ciprofloxacin, its gram-positive antimicrobial activity is marginal. The newer quinolones have added gram-positive activity, but little is known about cyst penetration by these antibiotics. We simultaneously measured cyst and serum levels of levofloxacin and ampicillin and found levofloxacin to penetrate cysts well. Therefore, levofloxacin may be useful in the management of renal cyst infection, particularly for the treatment of gram-positive organisms. PMID- 16377379 TI - Timed urine collections are not needed to measure urine protein excretion in clinical practice. PMID- 16377380 TI - Timed urine collections are not needed to measure urine protein excretion in clinical practice. PMID- 16377381 TI - Serial ANCA determinations for monitoring disease activity in patients with ANCA associated vasculitis: systematic review. AB - BACKGROUND: Antineutrophil cytoplasmic antibodies (ANCAs) are considered by some investigators to be sensitive markers of disease activity and have been suggested to predict relapse and guide therapeutic decisions. Studies using serial ANCA monitoring in patients with ANCA-associated vasculitis (AASV) have yielded controversial results during the last 15 years. To assess the diagnostic value of serial ANCA testing in the follow-up of patients with AASV, we conducted a systematic review of the available literature. METHODS: Studies were identified by a comprehensive search of the PubMed and BIOSIS+/RRM databases, as well as hand searching. Method quality of all eligible studies was assessed with respect to external and internal validity according to established criteria for diagnostic studies. RESULTS: Twenty-two studies met our inclusion criteria, including a total of 950 patients. Whereas generalizability was not a major problem, assessment of internal validity showed that only a minority of studies reported the combination of consecutive patient recruitment, prospective data collection, and independent determination of both index and reference tests, considered as the ideal for diagnostic test studies. Quantitative meta-analytic calculations were not conducted because of the presence of considerable method heterogeneity. CONCLUSION: The presence of considerable methodological heterogeneity combined with methodological shortcomings with respect to internal validity in the majority of included studies preclude firm conclusions from the available literature concerning the clinical value of serial ANCA determinations for monitoring the follow-up of patients with AASV. PMID- 16377382 TI - Evidence-based systematic literature review of hemoglobin/hematocrit and all cause mortality in dialysis patients. AB - BACKGROUND: An evidence-based evaluation of peer-reviewed original research published during 1980 to 2004 and examining the relationship between hemoglobin and/or hematocrit values and all-cause mortality in dialysis patients was conducted to compare the studies' designs, analytic strategies, and results. METHODS: The search targeted MEDLINE and EMBASE and included publications referenced in the National Kidney Foundation-Kidney Disease Outcomes Quality Initiative Anemia Guidelines. Both randomized clinical trials (RCTs) and observational studies were considered. RESULTS: Of 7 RCTs and 20 observational studies identified, 5 trials and 13 studies were included in evidence tables. The trials were underpowered to study mortality and enrolled different patient populations, limiting their generalizability. Although none reached statistical significance, trials focusing on a general dialysis population tended to show either no effect or a benefit of greater hemoglobin level target, whereas trials enrolling cardiac patients suggested increased mortality associated with greater hemoglobin levels. The observational studies were heterogeneous in design, used varying exposure categorizations, and controlled for different covariates, but generally were supportive of increased mortality associated with a hemoglobin level less than the reference range. Evidence of benefits or risks of hemoglobin levels greater than the reference was variable. CONCLUSION: RCT-based evidence relating hemoglobin and/or hematocrit values to mortality in dialysis patients is limited. The relationship may be modified by the presence of preexisting conditions (cardiac disease). The published literature is insufficient for generalization of risks or benefits of a hemoglobin level greater than 11 to 12 g/dL (>110 to 120 g/L). There is a need for better designed RCTs focusing on mortality as a primary outcome and enrolling patients without cardiac disease. Observational studies should adequately control for relevant confounders (eg, baseline comorbidities) and assess effect modification in the analysis. PMID- 16377383 TI - Myeloperoxidase as a marker of hemodialysis biocompatibility and oxidative stress: the underestimated modifying effects of heparin. AB - Myeloperoxidase (MPO) is a microbicidal and reactive species-generating enzyme. It traditionally is considered to be stored mostly within polymorphonuclear leukocytes and is strongly implicated in the pathogenesis of numerous diseases. MPO also has been studied for at least 20 years as a marker of hemodialysis procedure biocompatibility and oxidative stress generation; research yielded discordant and inconclusive results. In this review, a novel and growing body of evidence indicating that MPO also is a potent blood vessel-bound enzyme that can be mobilized rapidly and extensively into circulating blood by exogenous heparin is discussed. Beneficial consequences of such evoked arterial wall MPO depletion that may be counterbalanced in part by the harmful effects of circulating MPO on polymorphonuclear leukocyte activation and thus atherosclerosis propagation also are presented. Potential clinical implications of these undervalued phenomena in commonly atherosclerotic maintenance hemodialysis patients regularly administered large doses of heparin for temporary blood anticoagulation (frequently over years) are stressed, including the challenging issue of morbidity and mortality. In view of the plausible clinical importance of the novel MPO-oxidative stress heparin interaction in this population, the need for additional studies assessing different dialyzer membranes, various heparin types (unfractionated heparin versus low-molecular-weight heparins versus pentasaccharides), as well as different anticoagulation regimens, is emphasized. PMID- 16377384 TI - The determinants of endothelial dysfunction in CKD: oxidative stress and asymmetric dimethylarginine. AB - BACKGROUND: Oxidative stress is related to endothelial dysfunction (ED) and cardiovascular outcomes in patients with chronic kidney disease. Increased asymmetric dimethylarginine (ADMA) levels are among the main causes of ED. We aim to investigate any association between ED and ADMA levels, as well as levels of oxidative stress markers, in patients with chronic kidney disease. METHODS: One hundred fifty-nine patients without diabetes with chronic kidney disease were studied. Staging was performed according to glomerular filtration rate, determined as stages 1 to 5 according to the Kidney Disease Outcomes Quality Initiative (n = 30, 33, 28, 32, and 36, respectively). The control group consisted of 30 healthy subjects. Oxidative stress markers (plasma malondialdehyde [MDA], erythrocyte superoxide dismutase [SOD], glutathione peroxidase [GSH-Px]), trace elements (erythrocyte zinc [EZn], erythrocyte copper [ECu]), plasma selenium (Se), and serum ADMA were studied. Brachial artery endothelium-dependent vasodilatation (FMD) was calculated for all. RESULTS: FMD, SOD, GSH-Px, EZn, ECu, and Se values were lower, whereas MDA and ADMA levels were higher in patients than controls. Glomerular filtration rate correlated negatively with MDA and ADMA levels and positively with FMD, SOD, and GSH-Px values. These parameters were significantly different among patients with stages 2, 3, 4, and 5 (hemodialysis group; P < 0.001 for all). Regression analysis showed that ADMA (beta = -0.228; P < 0.01), SOD (beta = 0.405; P < 0.001), and oxidized low-density lipoprotein levels (beta = -0.428; P < 0.001) were related independently to FMD, whereas glomerular filtration rate was not involved in the model. CONCLUSION: The present results imply that FMD, oxidative stress, and ADMA levels all are associated with stage of chronic kidney disease. Additionally, levels of oxidative stress markers and ADMA independently determine endothelial function. PMID- 16377385 TI - Use of allopurinol in slowing the progression of renal disease through its ability to lower serum uric acid level. AB - BACKGROUND: Hyperuricemia is associated strongly with the development of hypertension, renal disease, and progression. Allopurinol decreases serum uric acid levels by inhibiting the enzyme xanthine oxidase. We hypothesized that administrating allopurinol to decrease serum uric acid levels to the normal range in hyperuricemic patients with chronic kidney disease may be of benefit in decreasing blood pressure and slowing the rate of renal disease progression in these patients. METHODS: We conducted a prospective, randomized, controlled trial of 54 hyperuricemic patients with chronic kidney disease. Patients were randomly assigned to treatment with allopurinol, 100 to 300 mg/d, or to continue the usual therapy for 12 months. Clinical, hematologic, and biochemical parameters were measured at baseline and 3, 6, and 12 months of treatment. We define our study end points as: (1) stable kidney function with less than 40% increase in serum creatinine level, (2) impaired renal function with creatinine level increase greater than 40% of baseline value, (3) initiation of dialysis therapy, and (4) death. RESULTS: One patient in the treatment group dropped out because of skin allergy to allopurinol. Serum uric acid levels were significantly decreased in subjects treated with allopurinol, from 9.75 +/- 1.18 mg/dL (0.58 +/- 0.07 mmol/L) to 5.88 +/- 1.01 mg/dL (0.35 +/- 0.06 mmol/L; P < 0.001). There were no significant differences in systolic or diastolic blood pressure at the end of the study comparing the 2 groups. There was a trend toward a lower serum creatinine level in the treatment group compared with controls after 12 months of therapy, although it did not reach statistical significance (P = 0.08). Overall, 4 of 25 patients (16%) in the allopurinol group reached the combined end points of significant deterioration in renal function and dialysis dependence compared with 12 of 26 patients (46.1%) in the control group (P = 0.015). CONCLUSION: Allopurinol therapy significantly decreases serum uric acid levels in hyperuricemic patients with mild to moderate chronic kidney disease. Its use is safe and helps preserve kidney function during 12 months of therapy compared with controls. Results of this study need to be confirmed with an additional prospective trial involving a larger cohort of patients to determine the long term efficacy of allopurinol therapy and in specific chronic kidney disease subpopulations. PMID- 16377386 TI - Plasma reduced homocysteine and other aminothiol concentrations in patients with CKD. AB - BACKGROUND: Hyperhomocysteinemia, a risk factor for cardiovascular disease, is present in the majority of patients with chronic kidney disease (CKD). Several studies indicated that the moiety of homocysteine (Hcy) with an unbound -SH group (reduced Hcy [rHcy]) is the atherogenic molecule. This study is designed to examine the relation between different forms of Hcy and other aminothiols in hemodialysis (HD) patients, peritoneal dialysis (PD) patients, and nondialyzed patients with CKD. METHODS: rHcy, free Hcy (fHcy), and total Hcy (tHcy), as well as different forms of cysteine, cysteinyl-glycine, and glutathione, were studied by using a high-performance liquid chromatography technique in 19 HD patients, 12 PD patients, 47 patients with CKD, and 15 control subjects. RESULTS: In PD patients, tHcy levels were 2.8 times greater compared with controls, and in HD patients and those with CKD, 2.1 and 1.9 times greater, respectively. Mean rHcy/tHcy ratios were significantly greater in both HD (P < 0.05) and PD patients (P < 0.01), but did not differ in patients with CKD compared with controls. The decrease in rHcy levels during 1 HD treatment was smaller than that in tHcy and fHcy levels, and rHcy/tHcy ratio increased (before HD, 1.25% +/- 0.44%; after HD, 1.44% +/- 0.66%; P < 0.05). CONCLUSION: Levels of rHcy and other aminothiols are markedly increased in patients with impaired renal function. In dialysis patients, rHcy/tHcy ratio is markedly elevated and shows greater variability than in patients with CKD and controls. We conclude that because rHcy is believed to induce endothelial dysfunction and may be part of the accelerated atherogenic process in patients with CKD, plasma rHcy level could be a more relevant marker of cardiovascular disease risk than tHcy level. PMID- 16377387 TI - CKD risk factors reported by primary care physicians: do guidelines make a difference? AB - BACKGROUND: End-stage renal disease is epidemic within the United States among certain high-risk groups. The National Kidney Disease Education Program examined the awareness of chronic kidney disease (CKD) risk factors among primary care physicians who treat high-risk populations, such as African Americans, persons with diabetes, persons with hypertension, and family members of dialysis patients. METHODS: We conducted a survey of 465 primary care physicians in 4 communities with high-risk populations. Respondents were asked to score 9 potential CKD risk factors on a 4-point scale graded from "Does not increase risk at all" to "Increases risk greatly." Potential risk factors included African American race, diabetes, hypertension, and family history of CKD. RESULTS: Respondents saw a mean of 414 +/- 222 (SD) patients/mo. Primary care physicians were more likely to report that diabetes and hypertension were significant risk factors for CKD. Conversely, 34.4% did not consider family history of kidney disease to increase the risk for CKD, and 22% of respondents did not consider African-American race a CKD risk factor. CONCLUSION: Primary care physicians need targeted education to increase awareness of populations at high risk for CKD. PMID- 16377388 TI - Renal function preservation in type 2 diabetes mellitus patients with early nephropathy: a comparative prospective cohort study between primary health care doctors and a nephrologist. AB - BACKGROUND: Early referral of patients with end-stage renal disease to a nephrologist is associated with lower morbidity and mortality after initiating dialysis therapy; earlier referral may have better results. The aim of the study is to prospectively determine the impact of earlier referral to a nephrologist on renal damage progression of patients with type 2 diabetes mellitus (DM2) with early nephropathy. METHODS: Fifty-two patients (27 patients, early nephropathy [EN]; 25 patients, overt nephropathy [ON]) from a primary health care unit were referred to a nephrologist (study cohort); 65 patients (34 patients, EN; 31 patients, ON) from another health care unit remained treated by only family doctors (control cohort). Both cohorts were followed up for 1 year. RESULTS: Delta (final-baseline) in serum creatinine levels was maintained better by the nephrologist in the EN (study, 0.02 mg/dL versus control, 0.13 mg/dL [2 versus 11 micromol/L]; P = 0.02) than ON group (study, 0.15 mg/dL versus control, 0.25 mg/dL [13 versus 22 micromol/L]). In concordance, glomerular filtration rate was maintained better by the nephrologist in EN (study, 3.2 mL/min/1.73 m2 versus control, -13.3 mL/min/1.73 m2 [0.05 versus -0.22 mL/s/1.73 m2]; P = 0.01) than ON patients (study, -9.8 mL/min/1.73 m2 versus control, -10.9 mL/min/1.73 m2 [-0.16 versus -0.18 mL/s/1.73 m2]). Albuminuria increased more in patients treated by family doctors in the EN (study, 30 mg/d versus control, 116 mg/d; P < 0.05) and ON groups (study, 160 mg/d versus control, 623 mg/d). The nephrologist controlled systolic blood pressure better in both the EN (study, -3 mm Hg versus control, 2 mm Hg; P < 0.05) and ON groups (study, -19 mm Hg versus control, 5 mm Hg; P < 0.05); diastolic blood pressure had a similar pattern. The nephrologist significantly increased (P < 0.05) the use of angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, and statins and discontinued nonsteroidal anti-inflammatory drugs more than family doctors (study, 42%, 43%, 39%, and -11% versus control, 17%, 4%, -7%, and 27%, respectively). Glycemic and lipid control and stopping smoking were not attained by either the nephrologist or family doctors. CONCLUSION: Earlier referral of patients with DM2 to a nephrologist was associated with better renal function preservation, which was significantly more evident in the EN than ON group. The nephrologist obtained better blood pressure control, more frequently used angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, and statins; and discontinued nonsteroidal anti-inflammatory drugs more than family doctors. However, metabolic control and stopping smoking were not attained by either the nephrologist or family doctors. PMID- 16377389 TI - Absence of HIV-associated nephropathy in Ethiopians. AB - BACKGROUND: Population-based epidemiological surveys in several countries have shown approximately 10- to 15-fold increased susceptibility to human immunodeficiency virus (HIV)-associated nephropathy (HIVAN) for populations of recent African ancestry. Accordingly, we sought to determine whether a similar or different pattern of susceptibility was evident among Ethiopians followed up in an HIV clinic in Israel. METHODS: One hundred seventy-six consecutive patients (126 Ethiopians, 50 non-Ethiopian Israelis) followed up at the HIV clinic of Rambam Medical Center in northern Israel were examined for the presence of proteinuria and/or decreased glomerular filtration rate. HIV viral load, CD4 count, and treatment modality also were determined. RESULTS: Overall, 73% of patients were treated with highly active antiretroviral therapy, and there was no difference between Ethiopians and non-Ethiopian Israelis in this regard. Mean CD4 count in Ethiopians was 288 +/- 140/microL, significantly less than the corresponding CD4 count of 398 +/- 190/microL for non-Ethiopian Israelis. Mean viral loads were greater in Ethiopians compared with non-Ethiopian Israelis. None of 176 HIV-infected patients fulfilled clinical criteria for HIVAN as delineated in this study. CONCLUSION: HIV-infected individuals of Ethiopian descent have a level of susceptibility to HIVAN similar to that of non-Ethiopian Israelis, which is strikingly less than that reported for other populations for recent African ancestry. This does not appear to be attributable to differences in HIV infection control or viral subtype and most likely represents population-based differences in host genetic factors. This finding emphasizes the importance of avoiding generalizations with respect to phylogeographic ancestry in disease susceptibility studies. PMID- 16377390 TI - Embolization of renal angiomyolipomata in patients with tuberous sclerosis complex. AB - BACKGROUND: Renal angiomyolipomata can reduce renal reserve and lead to renal insufficiency and failure. Angiomyolipomata often have abnormal vasculature, with aneurysms that can hemorrhage. Treatment of angiomyolipomata greater than 4 cm in diameter is suggested to decrease the risk for hemorrhage. Nephron-sparing procedures are critical in patients because of their limited renal reserve. Embolization has been used to treat these tumors, but there are limited studies examining efficacy. Our study examines the efficacy of selective embolization in decreasing tumor burden, preventing hemorrhage, and preserving renal function. METHODS: We conducted a retrospective study of 16 patients with 20 angiomyolipomata on 18 kidneys who underwent 18 transcatheter transarterial embolization procedures. Aneurysm number and size were documented and tumor volumes were measured before and after embolization. Preprocedure and follow-up renal function also were measured. Changes in angiomyolipoma volume and kidney function were assessed for significance by using paired t-test. RESULTS: Before embolization, 7 angiomyolipomata had more than 5 aneurysms, 9 had 1 to 5 aneurysms, and 4 had no aneurysms, but showed tortuous dysmorphic arteries. Mean aneurysm size was 5 mm. In patients available for follow-up, 15 of 16 tumors had decreased in volume (mean decrease, 56.1%; P = 0.001). At an average of 40 months' follow-up, there have been no subsequent hemorrhages. Patients' decline in renal function was not significantly different from that expected because of the natural course of the disease. CONCLUSION: Selective embolization decreases tumor size, prevents hemorrhage, and preserves kidney function in patients with tuberous sclerosis with renal angiomyolipomata. PMID- 16377391 TI - An evaluation of a shared primary and secondary care nephrology service for managing patients with moderate to advanced CKD. AB - BACKGROUND: Chronic kidney disease (CKD) is common, and nephrology services may not cope with the comprehensive referral of patients with CKD. We evaluated a shared primary and secondary care nephrology scheme, hypothesizing that some patients with less progressive moderate to advanced CKD can be identified and safely managed without attending the renal unit. METHODS: A retrospective review of 949 new referrals with stages 3 to 5 CKD managed in either the hospital nephrology clinic (HC) or the shared care scheme (SCS), in which nephrologists review patients remotely by using regular biochemical tests and clinical data recorded in primary care. RESULTS: Two hundred sixty-six patients (28%) were enrolled in the SCS and 683 patients (72%) were managed solely in the HC. Median time to entering the SCS was 111 days (interquartile range, 0 to 328 days). Baseline factors independently predictive of enrollment in the SCS were increasing age, greater glomerular filtration rate (GFR) and serum albumin levels, and no diabetic nephropathy. Few SCS patients did not attend reviews. Forty-one patients (15%) required recall to the HC, mostly because of a decline in GFR. Beneficial changes were seen in blood pressure levels and prescribing of angiotensin-system inhibitors from first referral to 3 years in all patients. Those enrolled in the SCS had good prognosis, with a lower risk for death or renal replacement therapy than the HC group after adjustment for age, sex, GFR, diabetic nephropathy, and vascular disease (hazard ratio, 0.64; 95% confidence interval, 0.38 to 0.89; P = 0.003). CONCLUSION: In this setting, it was possible to select nearly 30% of patients with stages 3 to 5 CKD for management in the SCS. More than half enrolled within 4 months of nephrology referral. Systematic surveillance was effective, and most patients remained stable, with few progressing to renal replacement therapy or death. PMID- 16377392 TI - Impact of specialization of primary nephrologist on the care of pediatric hemodialysis patients. AB - BACKGROUND: Children with end-stage renal disease (ESRD) receiving hemodialysis may have their care overseen primarily by a pediatric nephrologist or internal medicine (IM) nephrologist. METHODS: To examine specific clinical outcomes by nephrologist specialization, a cross-sectional analysis of demographic and clinical data collected in the 2002 ESRD Clinical Performance Measures Project was performed. RESULTS: Of 653 pediatric patients meeting inclusion criteria, 27% were cared for by IM nephrologists, and 73%, by pediatric nephrologists. Pediatric nephrologists were significantly more likely than IM nephrologists to care for patients who were younger and of Hispanic ethnicity. Patients of pediatric compared with IM nephrologists also were more likely to have a congenital cause of ESRD, smaller body mass index, and longer time on dialysis therapy. No significant differences in achieving a mean Kt/V of 1.2 or greater or mean hemoglobin level of 11 g/dL or greater (> or =110 g/L) according to nephrologist specialization were observed. After adjustment for patient clinical characteristics, no significant difference in use of arteriovenous fistulae was observed. Patients cared for by pediatric nephrologists were less likely to achieve a mean serum albumin level of 4.0/3.7 g/dL (40/37 g/L; bromcresol green laboratory method/bromcresol purple laboratory method; adjusted odds ratio, 0.60; 95% confidence interval, 0.42 to 0.86). Patients cared for by pediatric nephrologists had significantly greater serum calcium levels, lower serum phosphate levels, and lower intact parathyroid hormone levels. CONCLUSION: Using adult-focused clinical care targets, care provided by pediatric and IM nephrologists to pediatric patients receiving hemodialysis in the United States is similar. However, differences exist, and the significance of these differences requires further study. PMID- 16377393 TI - Families' and physicians' predictions of dialysis patients' preferences regarding life-sustaining treatments in Japan. AB - BACKGROUND: Substituted judgment traditionally has been used often for patient care in Japan regardless of the patient's competency. It has been believed that patient preferences are understood intuitively by family and caregivers. However, there are no data to support this assumption. METHODS: A questionnaire survey was administered to 450 dialysis patients in 15 hospitals to determine their preferences for cardiopulmonary resuscitation (CPR) and dialysis therapy under various circumstances. Simultaneously, we asked family members and physicians of these patients about patient preferences to evaluate their ability to predict what their patients would want. The accuracy of families' and physicians' judgments was assessed by means of kappa coefficient. RESULTS: Three hundred ninety-eight pairs, consisting of a patient, 1 of his or her family members, and the physician in charge, participated from 15 hospitals in Japan, with a response rate of 88%. Sixty-eight percent of family members correctly predicted patients' current preferences for CPR, 67% predicted patients' preferences for dialysis when they were severely demented, and 69% predicted patients' preferences for dialysis when they had terminal cancer. Corresponding figures for physicians were 60%, 68%, and 66%. When using kappa coefficient analysis, those results indicated that neither family members nor physicians more accurately predicted their patients' wishes about life-sustaining treatments than expected by chance alone. (All kappa coefficients <0.4.) CONCLUSION: Our study suggests that patients who want to spend their end-of-life period as they want should leave better advance directives. PMID- 16377394 TI - Cerebrovascular disease in maintenance hemodialysis patients: results of the HEMO Study. AB - BACKGROUND: Cerebrovascular disease (CBVD) in hemodialysis patients is associated with significant morbidity and mortality. A secondary analysis of CBVD in the Hemodialysis (HEMO) Study was performed. Specific objectives were to: (1) determine risk factors for the presence of CBVD at baseline, (2) assess risk factors for the subsequent occurrence of cerebrovascular deaths, and (3) analyze the effects of dose and flux on cerebrovascular mortality. METHODS: The HEMO Study was a randomized multicenter study evaluating the effects of high-dose versus standard-dose and high-flux versus low-flux hemodialysis. There were 1,846 patients enrolled, with a mean follow-up of 2.84 years. RESULTS: Factors associated with the baseline presence of CBVD included age (P < 0.0001), presence of any cardiac disease (P < 0.0001), and diabetes mellitus (P < 0.0001). There were 65 deaths caused by CBVD (event rate, 1.2/100 patient-years). Multivariate Cox regression using a backward-variables selection procedure showed that diabetes, lower albumin level, greater hematocrit, and lower body mass index at baseline were associated significantly with subsequent CBVD death. There was no effect of flux or dose on overall rate of CBVD deaths. However, an interaction was found between baseline CBVD status and flux intervention on CBVD death (P = 0.016). In the subgroup of patients without the baseline presence of CBVD, high flux dialysis was associated with a lower risk for death caused by CBVD (P = 0.016). A borderline interaction between years of dialysis therapy and flux on subsequent CBVD death was detected (P = 0.05). The beneficial effect of high flux was evident in those on hemodialysis therapy for longer than 3.7 years (P = 0.012). CONCLUSION: High flux was associated with decreased CBVD mortality in those without known CBVD at baseline and those on hemodialysis therapy for longer than 3.7 years. This secondary analysis strengthens, but does not prove, the hypothesis that high-flux treatment may attenuate the death rate from vascular disease. PMID- 16377395 TI - Serum albumin, C-reactive protein, interleukin 6, and fetuin a as predictors of malnutrition, cardiovascular disease, and mortality in patients with ESRD. AB - BACKGROUND: Serum albumin (S-Alb), C-reactive protein (CRP), and interleukin 6 (IL-6) predict malnutrition, atherosclerotic cardiovascular disease (CVD), and mortality in patients with end-stage renal disease (ESRD). Fetuin A, an inhibitor of vascular calcification, also is associated strongly with clinical outcome in patients with ESRD. In this study, multivariate analyses were performed to assess these 4 biomarkers as predictors of malnutrition, CVD, and mortality in patients with ESRD. METHODS: One hundred seventy-six patients with ESRD (54 +/- 12 years) underwent measurements of S-Alb, high-sensitivity CRP (hs-CRP), plasma IL-6, and fetuin A close to the start of dialysis therapy and were followed up for a median of 26 months (range, 1 to 66 months). Nutritional status was evaluated by means of subjective global assessment. CVD was defined based on medical history. Associations between biomarker levels and malnutrition, CVD, and mortality were evaluated by means of receiver operating characteristic curve, logistic regression analysis, and Cox proportional hazards model. RESULTS: All 4 biomarkers predicted malnutrition, CVD, and mortality. Multivariate analysis, according to receiver operating characteristic analysis, showed that malnutrition was predicted best by hs-CRP and IL-6 levels; CVD, by IL-6 level; and mortality, by albumin and IL-6 levels. When using the cutoff levels derived from receiver operating characteristics, logistic regression analysis showed that only hs-CRP level (odds ratio [OR], 3.6) was associated with malnutrition, and only IL-6 level (OR, 3.7) was associated with CVD. Levels of S-Alb, IL-6, and fetuin A, but not hs-CRP, were associated with increased relative risk for mortality as assessed by Cox in a model adjusting for age, sex, and diabetes mellitus. CONCLUSION: Multivariate analyses show that in patients with ESRD, malnutrition is predicted best by hs-CRP and IL-6 levels; CVD, by IL-6 level; and mortality, by S-Alb, IL-6, and fetuin A levels, but not by hs-CRP level. This comparative analysis indicates that of these biomarkers, IL-6 level may be the most reliable predictor of CVD and mortality in patients with ESRD. PMID- 16377396 TI - PTH and the risks for hip, vertebral, and pelvic fractures among patients on dialysis. AB - BACKGROUND: Few investigations have described fracture risk and its relation to disorders in calcium (Ca), phosphorus (P), and parathyroid hormone (PTH) metabolism in the end-stage renal disease population. METHODS: Laboratory values for Ca, P, and PTH were obtained from Dialysis Morbidity and Mortality Study (DMMS) Waves 1 to 4. Additional data available from the US Renal Data System were used to determine the incidence and associated costs of hip, vertebral, and pelvic fractures in 9,007 patients with nonmissing laboratory values and Medicare as primary payor. Cox proportional hazards and Poisson models were used to analyze time to first fracture and numbers of fractures, respectively. RESULTS: There was no association between Ca or P values and risk for fracture; risks for vertebral and hip fractures and PTH concentrations were U shaped and weakly significant using Poisson regression (P = 0.03). The age- and sex-adjusted mortality rate after fracture was 2.7 times greater (580/1,000 person-years) than for general dialysis patients from the DMMS (217/1,000 person-years). Mean total episodic costs of hip, vertebral, and pelvic fractures were 20,810 dollars +/- 16,743 dollars (SD), 17,063 dollars +/- 26,201 dollars, and 14,475 dollars +/- 19,209 dollars, respectively. CONCLUSION: Using data from the DMMS, there were no associations between Ca and P concentrations and risk for fracture. Risks for hip and vertebral fracture were associated weakly with PTH concentration, with the lowest risk observed around a PTH concentration of 300 pg/mL (ng/L). Fractures were associated with high subsequent mortality and costs. Prospective studies are needed to determine whether therapies that maintain PTH concentrations within or near the National Kidney Foundation-Kidney Disease Outcomes Quality Initiative range will result in fewer complications of disordered mineral metabolism. PMID- 16377397 TI - Challenging the validity of the EPO index. AB - BACKGROUND: With use of recombinant erythropoietin (EPO) and intravenous iron, the majority of hemodialysis patients can achieve target hemoglobin concentrations. EPO resistance arises as a consequence of inflammation and other processes that can adversely affect survival. We hypothesized that the EPO dose hematocrit (EPO/Hct) ratio, also known as the EPO index, may be a surrogate for inflammation and that greater EPO/Hct ratios would be associated with decreased survival. METHODS: We used proportional hazards regression models and time varying logistic models to analyze the association between EPO index and survival in US hemodialysis patients initiating hemodialysis therapy between January 1, 1999, and December 31, 2000, and followed up for up to 3 years until December 31, 2001. RESULTS: We found an unexpected and consistent association between greater EPO index and survival in all models. The associations of EPO/Hct ratio were most prominent at intermediate Hct values and with longer dialysis vintage. Iron administration was associated with a lower risk for death independent of Hct. Conversely, greater average prior EPO dose was associated with a greater risk for death. CONCLUSION: EPO resistance may be reflected better by total cumulative EPO dose than the EPO/Hct ratio. The mechanism(s) responsible for the association between a greater EPO/Hct ratio and survival remains to be established, but may be a result of nonerythrogenic effects of EPO. PMID- 16377398 TI - Human herpesvirus 8 infection in hemodialysis patients. AB - BACKGROUND: The aim of the present study was to evaluate human herpesvirus 8 (HHV 8) seroprevalence in Greek hemodialysis patients. Patterns of change in HHV-8 serostatus (seroconversions and seroreversions) over time were also evaluated. METHODS: Serum samples obtained from a cohort of 485 Greek hemodialysis patients were tested for antibodies to HHV-8 by whole virus lysate enzyme-linked immunosorbent assay, and reactive samples were confirmed by means of the orf-73 enzyme-linked immunosorbent assay. HHV-8 seroprevalence at study entry and the incidence of seroreversions and seroconversions per 100 person-years were estimated. RESULTS: The prevalence of HHV-8 antibodies in Greek hemodialysis patients at enrollment was 7.2%. No univariate associations were established between HHV-8 serostatus and patients' characteristics. Incidences of seroreversions and seroconversions were 16.4/100 person-years (95% confidence interval, 7.1 to 32.3) and 0.28/100 person-years (95% confidence interval, 0.03 to 1.02), respectively. Patients 50 years and younger had an increased probability for seroreversion to HHV-8 antibodies than patients older than 50 years (log-rank test, P = 0.018). CONCLUSION: We observed a fair number of seroreversions and a low incidence of seroconversion to HHV-8 infection in hemodialysis patients in Greece. Our data provide indirect evidence that HHV-8 transmission in the hemodialysis setting is uncommon. PMID- 16377399 TI - Target hemoglobin level for EPO therapy in CKD. PMID- 16377400 TI - Renal function testing. PMID- 16377401 TI - Evolution of lesions over 10 years in a patient with SLE: flowchart approach to the new International Society of Nephrology (ISN)/Renal Pathology Society (RPS) classification of lupus nephritis. PMID- 16377403 TI - If fat is good, muscle is better. PMID- 16377404 TI - Use of amino-terminal pro-B-type natriuretic peptide to predict coronary disease. PMID- 16377407 TI - Calcimimetics for predialysis patients? PMID- 16377408 TI - Radiation-enhanced immune response to cancer: workshop, Anaheim, CA, April 17, 2005. PMID- 16377409 TI - Pathogenic role of the renin-angiotensin system in modulating radiation-induced late effects. PMID- 16377411 TI - Combining radiotherapy and angiogenesis inhibitors: clinical trial design. AB - Radiotherapy (RT) plays a vital role in the multimodality treatment of cancer. Recent advances in RT have primarily involved improvements in dose delivery. Future improvements in tumor control and disease outcomes will likely involve the combination of RT with targeted therapies. Preclinical evaluations of angiogenesis inhibitors in combination with RT have yielded promising results with increased tumor "cure." It remains to be seen whether these improvements in tumor control in the laboratory will translate into improved outcomes in the clinic. Multiple differences between these agents and cytotoxic chemotherapy must be taken into account when designing clinical trials evaluating their effectiveness in combination with RT. We discuss important considerations for designing clinical trials of angiogenesis inhibitors with RT. PMID- 16377412 TI - Advances in understanding angiogenesis through molecular studies. AB - Tumors, in most cases, need angiogenesis for their sustained growth. A great deal of evidence has suggested that the process of angiogenesis is regulated by the balance between proangiogenic and antiangiogenic factors. Thus, the inhibition of tumor angiogenesis has been considered to be one of the key targets in anticancer therapy, and more than 60 antiangiogenic compounds are currently under clinical evaluation in cancer patients. However, the molecular mechanisms responsible for the activity of many of these antiangiogenic compounds are still not well understood. The recent development of microarray technology has allowed us to investigate the mechanism of action of these inhibitors more rapidly and extensively. With the use of microarray technology, novel molecules and pathways are shown to play a role in angiogenesis. This article also reviews new experimental approaches combined with microarray analysis to identify the molecular pathways involved in tumor-host interactions. Elucidation of the pathways that mediate both angiogenic and antiangiogenic responses will help us to develop better anticancer therapies. PMID- 16377413 TI - ZD6474, an inhibitor of VEGFR and EGFR tyrosine kinase activity in combination with radiotherapy. AB - Radiation enhances both epithelial growth factor receptor (EGFR) and vascular endothelial growth factor (VEGF) expression, which are a part of key pathways for tumor progression. Some tumors may not respond well to EGFR inhibitors alone or may develop resistance to EGFR inhibitors. Therefore, drug therapy targeted to VEGF receptors and EGFRs, when combined with radiotherapy (RT), may improve tumor control and provide wider applicability. This article focuses on ZD6474, an inhibitor of EGFR and VEGF receptor signaling in combination with RT. We discuss preclinical and clinical studies with RT and inhibitors of VEGF or EGFR signaling first. We then address issues associated with ZD6474 pharmacokinetic dosing, and scheduling when combined with RT. We also discuss ZD6474 in the context of anti EGFR therapy resistance. Dual inhibition of EGFR and VEGF receptor signaling pathways shows promise in enhancing RT efficacy. PMID- 16377414 TI - Molecular strategies targeting the host component of cancer to enhance tumor response to radiation therapy. AB - The tumor microenvironment, in particular, the tumor vasculature, as an important target for the cytotoxic effects of radiation therapy is an established paradigm for cancer therapy. We review the evidence that the phosphoinositide 3-kinase (PI3K)/Akt pathway is activated in endothelial cells exposed to ionizing radiation (IR) and is a molecular target for the development of novel radiation sensitizing agents. On the basis of this premise, several promising preclinical studies that targeted the inhibition of the PI3K/Akt activation as a potential method of sensitizing the tumor vasculature to the cytotoxic effects of IR have been conducted. An innovative strategy to guide cytotoxic therapy in tumors treated with radiation and PI3K/Akt inhibitors is presented. The evidence supports a need for further investigation of combined-modality therapy that involves radiation therapy and inhibitors of PI3K/Akt pathway as a promising strategy for improving the treatment of patients with cancer. PMID- 16377415 TI - Chemotherapy in locally advanced nasopharyngeal carcinoma: an individual patient data meta-analysis of eight randomized trials and 1753 patients. AB - OBJECTIVES: To study the effect of adding chemotherapy to radiotherapy (RT) on overall survival and event-free survival for patients with nasopharyngeal carcinoma. METHODS AND MATERIALS: This meta-analysis used updated individual patient data from randomized trials comparing chemotherapy plus RT with RT alone in locally advanced nasopharyngeal carcinoma. The log-rank test, stratified by trial, was used for comparisons, and the hazard ratios of death and failure were calculated. RESULTS: Eight trials with 1753 patients were included. One trial with a 2 x 2 design was counted twice in the analysis. The analysis included 11 comparisons using the data from 1975 patients. The median follow-up was 6 years. The pooled hazard ratio of death was 0.82 (95% confidence interval, 0.71-0.94; p = 0.006), corresponding to an absolute survival benefit of 6% at 5 years from the addition of chemotherapy (from 56% to 62%). The pooled hazard ratio of tumor failure or death was 0.76 (95% confidence interval, 0.67-0.86; p < 0.0001), corresponding to an absolute event-free survival benefit of 10% at 5 years from the addition of chemotherapy (from 42% to 52%). A significant interaction was observed between the timing of chemotherapy and overall survival (p = 0.005), explaining the heterogeneity observed in the treatment effect (p = 0.03), with the highest benefit resulting from concomitant chemotherapy. CONCLUSION: Chemotherapy led to a small, but significant, benefit for overall survival and event-free survival. This benefit was essentially observed when chemotherapy was administered concomitantly with RT. PMID- 16377416 TI - Plasminogen activator inhibitor-I-related regulation of procollagen I (alpha1 and alpha2) by antitransforming growth factor-beta1 treatment during radiation impaired wound healing. AB - PURPOSE: Plasminogen activator inhibitor (PAI)-1 mediates transforming growth factor-beta1 (TGF-beta1)-related signaling by stimulating collagen Type I synthesis in radiation-impaired wound healing. The regulation of alpha(I) procollagen is contradictory in fibroblasts of different fibrotic lesions. It is not known whether anti-TGF-beta1 treatment specifically inhibits alpha(I) procollagen synthesis. We used an experimental wound healing study to address anti-TGF-beta1-associated influence on alpha(I)-procollagen synthesis. METHODS AND MATERIALS: A free flap was transplanted into the preirradiated (40 Gy) or nonirradiated neck region of Wistar rats: Group 1 (n = 8) surgery alone; Group 2 (n = 14) irradiation and surgery; Group 3 (n = 8) irradiation and surgery and anti-TGF-beta1 treatment. On the 14th postoperative day, skin samples were processed for fibroblast culture, in situ hybridization for TGF-beta1, immunohistochemistry, and immunoblotting for PAI-1, alpha1/alpha2(I)-procollagen. RESULTS: Anti-TGF-beta1 significantly reduced TGF-beta1 mRNA (p < 0.05) and PAI-1 expression (p < 0.05). Anti-TGF-beta1 treatment in vivo significantly reduced alpha1(I)-procollagen protein (p < 0.05) and the number of expressing cells (p < 0.05) in contrast to significantly increased (p < 0.05) alpha2(I)-procollagen expression. CONCLUSION: These results emphasize anti-TGF-beta1 treatment to reduce radiation-induced fibrosis by decreasing alpha1(I)-procollagen synthesis in vivo. alpha1(I)-procollagen and alpha2(I)-procollagen might be differentially regulated by anti-TGF-beta1 treatment. Increased TGF-beta signaling in irradiated skin fibroblasts seemed to be reversible, as shown by a reduction in PAI-1 expression after anti-TGF-beta1 treatment. PMID- 16377417 TI - Evaluation of image-guided radiation therapy (IGRT) technologies and their impact on the outcomes of hypofractionated prostate cancer treatments: a radiobiologic analysis. AB - PURPOSE: To quantify the mitigation of geometric uncertainties achieved with the application of various patient setup techniques during the delivery of hypofractionated prostate cancer treatments, using tumor control probability (TCP) and normal tissue complication probability. METHODS AND MATERIALS: Five prostate cancer patients with approximately 16 treatment CT studies, taken during the course of their radiation therapy (77 total), were analyzed. All patients were planned twice with an 18 MV six-field conformal technique, with 10- and 5-mm margin sizes, with various hypofractionation schedules (5 to 35 fractions). Subsequently, four clinically relevant patient setup techniques (laser guided and image guided) were simulated to deliver such schedules. RESULTS: As hypothesized, the impact of geometric uncertainties on clinical outcomes increased with more hypofractionated schedules. However, the absolute gain in TCP due to hypofractionation (up to 21.8% increase) was significantly higher compared with the losses due to geometric uncertainties (up to 8.6% decrease). CONCLUSIONS: The results of this study suggest that, although the impact of geometric uncertainties on the treatment outcomes increases as the number of fractions decrease, the reduction in TCP due to the uncertainties does not significantly offset the expected theoretical gain in TCP by hypofractionation. PMID- 16377418 TI - Spinal cord planning risk volumes for intensity-modulated radiation therapy of head-and-neck cancer. AB - PURPOSE: To assess planning organ at risk volume (PRV) margins of the spinal cord in intensity-modulated radiotherapy (IMRT) of oropharyngeal cancers, by modeling the effect of geometric uncertainties to estimate the probability of the spinal cord receiving a particular dose. METHODS AND MATERIALS: Five patients with oropharyngeal cancer were treated by IMRT with simultaneous doses of 66 Gy (gross disease) and 54 Gy (subclinical disease) in 30 fractions. Spinal cord doses were limited to 45 Gy. The probability, due to random and systematic patient positioning uncertainties (3-mm standard deviation), of the cord receiving a particular dose was determined. The effect of an on-line setup correction protocol was also modeled. RESULTS: The mean probability of a maximum spinal cord dose of 45 Gy was 1%, with a 6-mm PRV margin. The mean probability of a maximum dose exceeding 40 Gy was 37% (range, 13-77%); this probability is reduced with a setup correction protocol. CONCLUSION: A spinal cord PRV generated with a 6-mm margin leads to a 99% probability of maintaining the maximum spinal cord dose below 45 Gy. The application of an on-line setup correction protocol reduces the cord dose by approximately 5 Gy. PMID- 16377420 TI - Serial megavoltage CT imaging during external beam radiotherapy for non-small cell lung cancer: in regard to Kupelian et al. (Int J Radiat Oncol Biol Phys 2005;630:1024-1028). PMID- 16377421 TI - Step-and-shoot IMRT vs. helical tomotherapy: in regard to van Vulpen et al. (Int J Radiat Oncol Biol Phys 2005;62;1535-1539). PMID- 16377422 TI - Large rectal cross section might be more than distention. PMID- 16377423 TI - Prostate-only not dead? PMID- 16377424 TI - Impairment in event sequencing in disorganised and non-disorganised patients with schizophrenia. AB - It has been suggested that thought disorder in schizophrenia is associated with impaired executive functions and with a defective understanding of others' behaviour. Action sequence knowledge processing and goal detection are considered as crucial components of executive functioning. Here, we used a picture sequencing task to assess the ability of schizophrenic patients (n = 40) with and without disorganisation symptoms to arrange different types of action sequence representations. Disorganisation symptoms appear to be associated with a general sequencing impairment, while patients without disorganisation symptoms displayed difficulties in ordering sequences requiring subjects to infer mental states in story characters along with a relatively preserved performance in correctly arranging mechanical or behavioural event sequences. These results reveal that only schizophrenic patients without disorganisation symptoms show a selective deficit in mentalising abilities whereas disorganisation symptoms are associated with a more severe event sequencing impairment probably reflecting basic failures of inferential reasoning. PMID- 16377425 TI - Functional frontoparietal connectivity during encoding and retrieval processes follows HERA model. A high-resolution study. AB - Recent neuroimaging studies of long-term episodic memory have suggested that left prefrontal cortex predominates in encoding condition, whereas right prefrontal cortex predominates in retrieval condition (hemispheric encoding and retrieval asymmetry, HERA model). The present electroencephalographic (EEG) study investigated the functional coupling of fronto-parietal regions during long-term memorization of visuo-spatial contents (i.e. landscapes, interiors of apartments), to test the predictions of the HERA model. Global fronto-parietal coupling was estimated by spectral coherence, whereas the "direction" of the fronto-parietal information flow was estimated by directed transfer function (DTF). The EEG rhythms of interest were theta (4-7 Hz), alpha (8-13 Hz), beta (14 30 Hz), and gamma (30-45 Hz). Statistically significant coherence in line with the HERA model was obtained at the gamma band. Namely, the fronto-parietal gamma coherence prevailed in the left hemisphere during the encoding condition and in the right hemisphere during the retrieval condition. The DTF estimates of the gamma band showed a dominant parietal-to-frontal directional flow in the right hemisphere during the encoding condition and in the left hemisphere during the retrieval condition (i.e. hemisphere-condition combination not involved by the HERA model). In contrast, a balanced bidirectional flow of the fronto-parietal coupling was observed in the left hemisphere during the encoding condition and in the right hemisphere during the retrieval condition (i.e. hemisphere-condition combination involved by the HERA model). In conclusion, the present encoding retrieval conditions induced maximal fronto-parietal gamma coupling with bidirectional information flow in the hemisphere-condition combination predicted by the HERA model. PMID- 16377426 TI - Antibody formation against beta-tubulin class III in response to brain trauma. AB - Brain trauma typically leads to neuronal damage and loss. Assuming a transient autoimmune response to debris of the damaged neurones, we have monitored serum titres of IgG and IgM antibodies to beta-tubulin class III (betaTcIII), which is almost exclusively found in neuronal cytoskeletons. In 15 out of 18 patients, the peak of the IgG or IgM antibody titre appeared in the serum within 3 weeks of a brain trauma. PMID- 16377427 TI - Relation between motion sickness susceptibility and vasovagal syncope susceptibility. AB - Motion sickness is common in the population, especially in children, but its physiopathology is only partially understood and the true nature of the particular susceptibility of certain subjects remains completely unknown. Some symptoms of motion sickness, like pallor and cold sweating, are of an autonomic nature and the role of the autonomic nervous system in vasovagal syncope is well known. Our aim was therefore to study the relationship between motion sickness susceptibility and vasovagal syncope susceptibility. Questionnaires about susceptibility to motion sickness and to vasovagal syncope or presyncope in adulthood and childhood, filled in by 899 students (20.4 +/- 2.1 years, 405 men), were analysed. Motion sickness susceptibility in childhood was 31.1% and in adulthood 7.9% (p < 0.001). Vasovagal syncope susceptibility in childhood was 36.4% and in adulthood 33.9% (NS). A relationship between motion sickness susceptibility in adulthood and vasovagal syncope susceptibility in childhood and adulthood (p = 0.004 and 0.005, respectively) was found. Despite the limitations of a retrospective study this relationship between motion sickness susceptibility and vasovagal syncope susceptibility may indicate that a common mechanism exists, explaining the particular susceptibility of some subjects to both disorders. This paradigm may prove useful in better understanding the true nature of motion sickness and vasovagal syncope. PMID- 16377428 TI - Occurrence of cognitive impairment after age 90: MCI and other broadly used concepts. AB - BACKGROUND: Few data are available on the impact of applying different concepts to define cognitive impairment among nonagenarians and centenarians. This study explored the occurrence of four broadly used concepts in an over 90-year-old population. DESIGN/METHODS: We examined a community-based cohort of 34 people with a mean age (+/- S.D.) of 96.4 (+/- 3.9) years, living in Bologna (Italy). We calculated the proportion of subjects that was given a diagnosis of cognitive impairment according to the following four commonly used concepts: Mild Cognitive Impairment (MCI), Age-Associated Memory Impairment (AAMI), Cognitive Impairment, No Dementia (CIND), and Questionable Dementia stage of the Clinical Dementia Rating (CDR 0.5). RESULTS: The proportion of subjects with cognitive impairment varied from 5.9%, according to CIND, to 32.4%, according to MCI. The four concepts identified different groups of subjects as having cognitive impairment, and no subject was given a diagnosis of cognitive impairment according to all four concepts. CONCLUSION: Commonly used criteria for diagnosis of cognitive impairment can differ by a factor of five in the number of subjects they classified as impaired. Such disagreement has serious implications for prevention, as people with a mild cognitive impairment may be a suitable target group for interventions before the development of dementia. PMID- 16377429 TI - Learning-dependent activation of Fra-1: involvement of ventral hippocampus and SNc/VTA complex in learning and habit formation. AB - Although the effect of overtraining on learning processes in rats has long been studied, only few studies have specifically assessed the differential involvement of brain areas in habit formation. We used the analysis of expression of the immediate early gene Fra-1 as a tool to differentiate the areas involved in training and overtraining. Behavioural experiments showed that instrumental performance (signalled and non-signalled instrumental tasks), but not pavlovian conditioned responses, were no longer under the control of the incentive value of the reward after overtraining. The number of Fra-1 expressing neurons was increased in SNc/VTA and ventral hippocampus after training in all groups independently of behavioural performance. After overtraining, the number of learning-induced Fra-1 immunoreactive neurons remained increased in the SNc/VTA. However, in CA1, it significantly decreased in the signalled instrumental group, whereas it further increased in the pavlovian group, with no modulation in non signalled instrumental animals. The increase in the number of Fra-1 neurons observed after training in SNc/VTA and ventral hippocampus suggests that a general underlying incentive process regulates Fra-1. Moreover, the sustained increased expression of Fra-1 in the SNc/VTA after instrumental overtraining could reflect a possible role of dopaminergic neurons in habit formation. PMID- 16377430 TI - Ultrastructural evidence for direct excitatory retroambiguus projections to cutaneous trunci and abdominal external oblique muscle motoneurons in the cat. AB - The nucleus retroambiguus (NRA) is a group of neurons, located laterally in the caudal medulla oblongata. The NRA is thought to modulate abdominal pressure in the framework of respiration, vomiting, vocalization, probably parturition, and, in all likelihood mating behavior. The NRA exerts this control through its projections to motoneurons to the nucleus ambiguus in the lateral medulla (innervating pharynx, larynx), and spinal cord (innervating cutaneous trunci, intercostal, abdominal, pelvic floor, and lower limb muscles). The nature of these NRA-motoneuronal projections is unknown. In this study we have determined the ultrastructure of the NRA-motoneuronal projections, and especially those to the abdominal external oblique and cutaneous trunci muscles. In four cats 0.1% cholera toxin subunit b was injected in the external oblique and cutaneous trunci muscles to retrogradely label their motoneurons in the spinal cord. Wheat germ agglutinin-conjugated horseradish peroxidase was injected into the NRA to anterogradely label its contralaterally descending fibers to the motoneurons of both muscles. In order to prevent anterograde labeling of ipsilaterally descending systems not originating from the NRA, a hemisection was made at the level of C2 prior to the NRA injection. The ultrastructural results indicate that the majority (60-74%) of the anterogradely labeled NRA-terminals made monosynaptic contacts with retrogradely labeled dendrites of the external oblique and the cutaneous trunci muscle motoneurons. The majority (86-95%) of the NRA terminals made asymmetric synaptic contacts and 79-84% contained round vesicles. These results demonstrate the existence of direct, presumably excitatory, projections from NRA to external oblique and cutaneous trunci muscle motoneurons. PMID- 16377431 TI - Nitric oxide synthesis is required for exercise-induced increases in hippocampal BDNF and phosphatidylinositol 3' kinase expression. AB - Previous studies have shown that running exercise, either alone or in combination with antidepressant treatment, results in increased hippocampal BDNF levels. Nitric oxide (NO) is an important signaling molecule that has neuronal survival promoting properties and has been shown to play an important role in plasticity associated with activating interventions. Herein, we administered the NO synthase (NOS) inhibitor, N-nitro-L-arginine methyl ester (L-NAME), in conjunction with the monoamine oxidase inhibitor (MAOI) antidepressant, tranylcypromine, and voluntary wheel-running exercise to determine whether the enhancement in full length BDNF mRNA occurring with these interventions is dependent upon NO synthesis. Our results demonstrate that both chronic exercise and chronic exercise-plus-tranylcypromine lead to enhanced hippocampal BDNF mRNA and protein expression. NOS inhibition prevents this effect of chronic exercise, but only partly prevents the effects of the exercise/antidepressant combination. Thus, the robust enhancement in BDNF mRNA occurring with exercise appears to be NO synthesis-dependent, but the intervention including antidepressant may enhance BDNF expression through alternative intracellular mechanisms. In addition, because exercise and antidepressants have both been shown to activate survival promoting genes, we evaluated the levels of hippocampal phosphatidylinositol 3' kinase (PI-3K), an important signaling molecule within a principal neuronal survival-promoting intracellular pathway. Like BDNF mRNA and protein, exercise increases the expression of PI-3K, whereas concomitant NOS inhibition prevents this increase in PI-3K immunoreactivity above control levels. Our results are discussed in light of possible overlapping, but distinct intracellular pathways activated by exercise and antidepressant treatment to bring about enhancements in BDNF expression and other survival-promoting effects. These findings further demonstrate the potential therapeutic potential of chronic exercise to supplement pharmacotherapeutic treatment of mood disorders. PMID- 16377432 TI - Talampanel improves the functional deficit after transient focal cerebral ischemia in rats. A 30-day follow up study. AB - The neuroprotective effect of talampanel, a negative allosteric modulator of alpha-amino-3-hydroxy-methyl-4-isoxazolyl-propionic acid (AMPA) receptors has been described previously. However, in these studies the histological changes and not the functional consequences of the brain damage were evaluated. The aim of present investigation was to analyze the sensorimotor function after stroke and to test the influence of talampanel (GYKI-53773, LY-300164) by 30-day monitoring in rats. After 1h middle cerebral artery occlusion (MCAO) general 'well-being', neurological status, spontaneous motor activity, rotation, motor coordination, balancing, muscle strength and reaction time were followed for 1 month. Talampanel (6 x 10 mg/kg i.p. given on the day of stroke) improved the motor coordination in rotarod (p < 0.01) and beam walking (p < 0.01) tests, reduced the number of stroke-induced rotations (p < 0.05), shortened the reflex time on the forelimb contralateral to brain ischemia and improved the survival rate comparing with vehicle treated control. After stroke, serious sensorimotor deficits appeared in rats but they showed partial spontaneous recovery after 30 days. Talampanel treatment enhanced the rate of functional improvement without changing the morphology at the end of the experiment. Our results indicate that modulation of AMPA receptors by talampanel can be a promising therapeutic approach to the treatment of stroke. PMID- 16377433 TI - Cocaine-induced sex differences in D1 receptor activation and binding levels after acute cocaine administration. AB - Although it is established that female rats have a more robust behavioral response to acute cocaine administration than male rats, the neurobiological mechanisms underlying these differences remain unclear. The purpose of the present study was to determine whether dopamine (DA) receptor activation influences sex differences in cocaine-induced behaviors. A second study was performed to determine sex differences in D1/D2 receptor levels prior to and post cocaine administration. Male and female Fischer rats were pre-treated with the D1 antagonist SCH-23390 (0.05, 0.1, and 0.25 mg/kg, i.p.), the D2 antagonist eticlopride (0.03, 0.1 mg/kg, i.p.), or vehicle (saline) 15 min before acute cocaine (20 mg/kg, i.p.) or saline administration. Cocaine-induced ambulatory and rearing activity was greater in female than male rats. Pre-treatment with SCH 23390 affected cocaine-induced ambulatory, rearing, and stereotypic activity in a sex-dependent manner; cocaine-induced ambulatory and stereotypic behavior in female rats was reduced by the lowest dose of SCH-23390. Eticlopride did not alter behavioral responses to cocaine in male or female rats. These results suggest that in both male and female rats, activation of the D1, but not the D2, receptor modulates cocaine's motor effects. There were no sex differences in baseline levels of D1, D2, and DA transporter binding in the caudate putamen (CPu) and the nucleus accumbens (NAc). Cocaine administration reduced D1 binding levels in the CPu only in male rats. Our findings suggest that the regulation of striatal D1 binding levels after acute cocaine administration is a sexually dimorphic process. We also hypothesize that the greater sensitivity to D1 receptor blockade in female rats, as compared to male rats, may contribute to their overall increased hyperactivity in response to acute cocaine. Taken together, the D1 receptor may be an important substrate in the regulation of sex differences to cocaine-induced locomotor activity. PMID- 16377434 TI - Visual responses and afferent connections of the n. ventrolateralis thalami (VLT) in the pigeon (Columba livia). AB - The nucleus ventrolateralis thalami (VLT) in pigeons receives direct retinal and forebrain projections and has reciprocal connections with the optic tectum. Although VLT is a component of the avian visual system, no study directly examined its connections or its cellular response characteristics. We, therefore, recorded from single units in the pigeon's VLT while visually stimulating the ipsi- and/or contralateral eye. In addition, tracing experiments were conducted to investigate its afferent connections. Electrophysiologically, we discovered three types of neurons, two of which were probably activated via a top-down telencephalotectal system (latencies > 100 ms). Type I neurons responded to uni- and bilateral and type II neurons exclusively to bilateral stimulation. Type III neurons were probably activated by retinal or retinotectal input (latencies < 27 ms) and responded to contra- and bilateral stimulation. Retrograde tracer injections into the VLT revealed an ipsilateral forebrain input from the visual Wulst, from subregions of the arcopallium, and bilateral afferents from the optic tectum. Most intriguing was the direct connection between the VLTs of both hemispheres. We suggest that the avian VLT is part of a system that integrates visuomotor processes which are controlled by both forebrain hemispheres and that VLT contributes to descending tectomotor mechanisms. PMID- 16377435 TI - Microinjection of procaine and electrolytic lesion in the ventral tegmental area suppresses hippocampal theta rhythm in urethane-anesthetized rats. AB - The midbrain ventral tegmental area (VTA), a key structure of the mesocorticolimbic system is anatomically connected with the hippocampal formation. In addition mesocortical dopamine was found to influence hippocampus related memory and hippocampal synaptic plasticity, both being linked to the theta rhythm. Therefore, the aim of the present study was to evaluate the possible role of the VTA in the regulation of the hippocampal theta activity. The study was performed on urethane-anesthetized male Wistar rats in which theta rhythm was evoked by tail pinch. It was found that unilateral, temporal inactivation of the VTA by means of direct procaine injection resulted in bilateral suppression of the hippocampal theta which manifested as a loss of synchronization of hippocampal EEG and respective reduction of the power and also the frequency of the 3-6 Hz theta band. Depression of the power of the 3-6 Hz component of the EEG signal was also seen in spontaneous hippocampal EEG after procaine. The permanent destruction of the VTA by means of unilateral electrocoagulation evoked a long-lasting, mainly ipsilateral depression of the power of the theta with some influence on its frequency. Simultaneously, there was a substantial increase of the power in higher frequency bands indicating decrease of a synchrony of the hippocampal EEG activity. On the basis of these results indicating impairment of synchronization of the hippocampal activity the VTA may be considered as another part of the brainstem theta synchroning system. PMID- 16377436 TI - Estrogen and progesterone affect cocaine pharmacokinetics in female rats. AB - Several studies have reported sex differences in behavioral responses to cocaine whereby females display a greater degree of locomotor activity. Fluctuations in estrogen and progesterone during the estrous cycle have been postulated to underlie these behavioral differences. In this study, we tested the hypothesis that hormonal replacement (estrogen or progesterone) in ovariectomized rats affects cocaine pharmacokinetics. We found that estrogen replacement did not affect cocaine-induced locomotor activity, but progesterone attenuated locomotor counts in comparison with control groups receiving only sesame oil. Estrogen, however, decreased brain levels of cocaine and norcocaine 30 min after cocaine administration in comparison to the group-receiving vehicle at that time point. In addition, in progesterone-treated rats, levels of benzoylecgonine and ecgonine methylester were higher at 30 min post-administration than at 15 min. No changes were found in blood levels of the metabolites. These findings suggest that while progesterone has an impact on locomotor behavior, pharmacokinetic effects may have a limited role in mediating behavioral responses to cocaine. PMID- 16377437 TI - Elucidating coding of taste qualities with the taste modifier miraculin in the common marmoset. AB - To investigate the relationships between the activity in different types of taste fibers and the gustatory behavior in marmosets, we used the taste modifier miraculin, which in humans adds a sweet taste quality to sour stimuli. In behavioral experiments, we measured marmosets' consumption of acids before and after tongue application of miraculin. In electrophysiological experiments responses of single taste fibers in chorda tympani and glossopharyngeal nerves were recorded before and after tongue application of miraculin. We found that after miraculin marmosets consumed acids more readily. Taste nerve recordings showed that after miraculin taste fibers which usually respond only to sweeteners, S fibers, became responsive to acids. These results further support our hypothesis that the activity in S fibers is translated into a hedonically positive behavioral response. PMID- 16377438 TI - Looming-sensitive responses and receptive field organization of telencephalic neurons in the pigeon. AB - The tectofugal pathway in birds goes from the optic tectum to the telencephalic entopallium via the thalamic nucleus rotundus (nRt). This pathway may be homologous to the colliculo-pulvinar-cortical pathway in mammals. It is known that a population of rotundal neurons in the pigeon can signal impending collision of looming objects with the animal. Here we show by single-unit recording that there exist two groups of looming-sensitive neurons in the entopallium. A tau cell starts firing at a nearly constant time before collision whereas the response onset time of an eta cell is linearly related to the square root of the diameter/velocity ratio of looming objects. These cells are localized in the caudal entopallium. The receptive field (RF) of looming-sensitive cells was mapped on the screen plane but its inhibitory region could not suppress responses to looming objects. It appears that a population of telencephalic cells in pigeons responds to looming objects and their looming responses are not determined by the receptive field organization mapped on the screen plane. PMID- 16377439 TI - Synthesis, conformational and pharmacological studies of glycosylated chimeric peptides of Met-enkephalin and FMRFa. AB - Our previous study showed that a chimeric peptide of Met-enkephalin and FMRFamide, YFa (YGGFMKKKFMRFa) not only caused antinociception and potentiated morphine analgesia but also blocked the development of tolerance and physical dependence. In the continuation of that study three chimeric analogues of YFa, [Ser5]YFa, [O-Glu-Ser5]YFa and [O-Gal-Ser5]YFa, were synthesized. To increase the bioavailability and penetration of blood brain barrier (BBB), glycosylated analogues, [O-Glu-Ser5]YFa and [O-Gal-Ser5]YFa, have been synthesized by solid phase peptide synthesis by building block method using anomeric acetate activation method. Circular dichroism studies showed that all the three chimeric peptides are stable and have a propensity for adopting helical conformation in the presence of membrane mimicking solvent. In comparison of parent chimeric peptide YFa, helicity of [Ser5]YFa, [O-Glu-Ser5]YFa and [O-Gal-Ser5]YFa has decreased. Pharmacological studies using tail-flick latency in mice showed that [O-Glu-Ser5]YFa have increased analgesia and bioavailability in comparison of [O Gal-Ser5]YFa and non-glycosylated analogue [Ser5]YFa. Exhibition of enhanced analgesia by [O-Glu-Ser5]YFa as compared to [O-Gal-Ser5]YFa seems to be due to preference of GLUT-1 transporter system for glucose. PMID- 16377440 TI - Enhanced central hypertonic saline-induced activation of angiotensin II-sensitive neurons in the anterior hypothalamic area of spontaneously hypertensive and Dahl S rats. AB - High dietary salt intake activates the brain renin-angiotensin system in spontaneously hypertensive rats (SHR) and Dahl S rats, resulting in sympathetic hyperactivity and hypertension. Increases of sodium concentration in cerebrospinal fluid (CSF) and/or enhanced responses to CSF sodium are considered to be involved in the high dietary salt-induced activation of central nervous system pathways in those rats. Previously we have demonstrated that intracerebroventricular injection of hypertonic saline increases the neural activity of angiotensin II-sensitive neurons trans-synaptically via endogenous angiotensins in the anterior hypothalamic area (AHA) of rats. In the present study, we examined whether the AHA angiotensin II-sensitive neuron response to hypertonic saline would differ in SHR and Dahl S rats from those of their controls. Male 15- to 16-week-old SHR and age-matched Wistar Kyoto rats (WKY), Dahl S rats and Dahl R rats and Wistar rats were anesthetized and artificially ventilated. Extracellular potentials were recorded from single neurons in the AHA. Intracerebroventricular injection of hypertonic saline increased the firing rate of AHA angiotensin II-sensitive neurons. The threshold sodium concentration for the central sodium-induced increase of neural firing was lower in SHR than those of WKY, Dahl S rats, Dahl R rats and Wistar rats. The increase in neural firing induced by hypertonic saline (250 mM) was greater in SHR than those of other four kinds of rats. Similarly, the threshold sodium concentration was lower in Dahl S rats than those of WKY, Dahl R rats and Wistar rats and the increase in neural firing induced by hypertonic saline (250 mM) was greater in Dahl S rats than those of WKY, Dahl R rats and Wistar rats. In SHR, intracerebroventricular injection of the amiloride-sensitive sodium channel blocker benzamil abolished the hypertonic saline (250 mM)-induced increase in neural firing, but the sodium channel blocker itself did not affect the basal firing of these neurons. These findings indicate that central sodium-induced activation of AHA angiotensin II sensitive neurons is enhanced in SHR and Dahl S rats. PMID- 16377441 TI - Cholinergic receptor subtypes in the olfactory bulbectomy model of depression. AB - The connection between smoking and depression, the antidepressant actions of nicotine and the targeting of nicotinic acetylcholine receptors (nAChRs) by monoamine re-uptake inhibitors all point to a potential role of nAChRs in the etiology and/or symptomatology of depression. In the current study, we evaluated nAChR subtypes in brain regions of rats subjected to olfactory bulbectomy (OBX), a standard animal model that recapitulates many of the behavioral and neurochemical alterations thought to underlie human depression. Comparisons were made both to sham-operated controls and unoperated animals. OBX led to upregulation of cerebrocortical alpha4beta2 nAChRs and downregulation of striatal alpha7 nAChRs as compared to either the sham-operated or unoperated groups. Striatal alpha4beta2 nAChRs were also downregulated but the sham surgery by itself produced a partial effect, masking the contribution of the OBX lesion. In agreement with earlier studies, we also found downregulation of muscarinic AChRs (both m1 and m2 subtypes) in the striatum when comparing the OBX group to sham operated controls, but because sham surgery evoked mAChR upregulation, the effect was not apparent when the OBX animals were contrasted to the unoperated group. Accordingly, caution needs to be exercised in interpreting studies of cholinergic function in the OBX model that do not include unoperated animals as an additional comparison group. Our results reinforce a relationship between depression and nAChR expression and point to the need for parallel studies in human depression that might lead to the design of novel therapies targeting specific nAChR subtypes. PMID- 16377442 TI - Correlative studies support lipid peroxidation is linked to PrP(res) propagation as an early primary pathogenic event in prion disease. AB - To assess whether heightened oxidative stress plays an early and primary pathogenic role in transmissible spongiform encephalopathies (TSE), we undertook detailed correlative studies using a mouse-adapted model of human disease. The spatio-temporal evolution of the abnormal, protease-resistant isoform of the prion protein (PrP(res)) and neuropathological changes were correlated with the occurrence and type of oxidative stress. Heightened oxidative stress was demonstrated, but restricted to elevated levels of free aldehydic breakdown products of lipid peroxidation, affecting all brain regions to varying extents. The increase in lipid peroxidation was highest over the mid-incubation period, with the onset showing close temporal and general topographical concordance with the first detection of PrP(res) with both pre-empting the typical neuropathological changes of spongiform change, gliosis and neuronal loss. Further, prion propagation over the disease course was assessed using murine bioassay. This revealed that the initial rapid increase in infectivity titres was contemporaneous with the abrupt onset and maximisation of lipid peroxidation. The present results are an important extension to previous studies, showing that heightened oxidative stress in the form of lipid peroxidation is likely to constitute an early primary pathogenic event in TSE, associated temporally with the integral disease processes of prion propagation and PrP(res) formation, and consistent with causal links between these events and subsequent typical neuropathological changes. PMID- 16377443 TI - Effects of extremely low-frequency magnetic field in the brain of rats. AB - An extremely low-frequency magnetic field (50 Hz, 0.5 mT) was used to investigate its possible effect on the brain of adult male Wistar rats following a 7-day exposure. The control rats were sham-exposed. Superoxide dismutase activities and production of superoxide radicals, lipid peroxidation, and nitric oxide were examined in the frontal cortex, striatum, basal forebrain, hippocampus, brainstem, and cerebellum. Significantly increased superoxide radical contents were registered in all the structures examined. Production of nitric oxide, which can oppose superoxide radical activities, was significantly increased in some structures: the frontal cortex, basal forebrain, hippocampus, and brainstem. Augmentation of lipid peroxydation was also observed, with significance only in the basal forebrain and frontal cortex, in spite of the significantly increased superoxide dismutase activities and nitric oxide production in the basal forebrain, and increased production of nitric oxide in the frontal cortex. The results obtained indicate that a 7-day exposure to extremely low-frequency magnetic field can be harmful to the brain, especially to the basal forebrain and frontal cortex due to development of lipid peroxidation. Also, high production of superoxide anion in all regions may compromise nitric oxide signaling processes, due to nitric oxide consumption in the reaction with the superoxide radical. PMID- 16377444 TI - Dopamine release in human neocortical slices: characterization of inhibitory autoreceptors and of nicotinic acetylcholine receptor-evoked release. AB - The autoinhibitory control of electrically evoked release of [3H]-dopamine and the properties of that induced by nicotinic receptor (nAChR) stimulation were studied in slices of the human neocortex. In both models [3H]-dopamine release was action potential-induced and exocytotic. The selective dopamine D2 receptor agonist (-)-quinpirole reduced electrically evoked release of [3H]-dopamine, yielding IC50 and I(max) values of 23 nM and 76%, respectively. Also, the effects of several other subtype-selective dopamine receptor ligands confirmed that the terminal dopamine autoreceptor belongs to the D2 subtype. The autoinhibitory feedback control was slightly operative under stimulation conditions of 90 pulses and 3 Hz, with a biophase concentration of endogenous dopamine of 3.6 nM, and was enhanced under blockade of dopamine reuptake. [3H]-dopamine release evoked in an identical manner in mouse neocortical slices was not inhibited by (-)-quinpirole, suggesting the absence of dopamine autoreceptors in this tissue and underlining an important species difference. Also, nAChR stimulation-induced release of [3H] dopamine revealed a species difference: [3H]-dopamine release was evoked in human, but not in rat neocortical slices. The nAChRs inducing [3H]-dopamine release most probably belong to the alpha3/beta2subtype, according to the potencies and efficacies of subtype-selective nAChR ligands. Part of these receptors may be located on glutamatergic neurons. PMID- 16377445 TI - The effect of leptin on penicillin-induced epileptiform activity in rats. AB - Leptin is an adipose tissue-derived peptide hormone, which acts as a satiety factor to reduce appetite by interactions with hypothalamic neurons. The other possible physiological functions of leptin are still unclear. In this study, we have evaluated dose-dependent effect of leptin on penicillin-induced epileptiform activity, analyzed by electrocorticogram (ECoG). The epileptiform activity was induced by microinjection of penicillin into the left sensorymotor cortex. Thirty minutes after penicillin injection, 1, 2 or 10 microg of leptin was administrated intracerebroventricularly (i.c.v.). Leptin (1, 2 or 10 microg) alone did not significantly change the spike amplitudes in non-penicillin pretreated control animals. One or two micrograms of leptin significantly increased the frequency of epileptiform activity in the penicillin-pretreated animals. The high dose of leptin (10 microg) did not significantly change either amplitude or frequency of epileptiform activity. One microgram i.c.v. leptin was the most effective dose in changing of frequency on penicillin-induced epileptiform activity. The proconvulsant effects of leptin appeared 90 min after leptin (1 and 2 microg) injection. These data indicate that leptin increases the frequency of penicillin induced epileptic activity. We speculate that this action of leptin might suggest that leptin may be a proconvulsant substance. PMID- 16377446 TI - Protective effect of S-allylcysteine on 3-nitropropionic acid-induced lipid peroxidation and mitochondrial dysfunction in rat brain synaptosomes. AB - 3-Nitropropionic acid is a neurotoxin that irreversibly inhibits succinate dehydrogenase, a relevant enzyme constituting the complex II of the respiratory chain during mitochondrial electron transport. 3-Nitropropionic acid is known to produce oxidative/nitrosative stress and evokes an experimental model of Huntington's disease. In this work we evaluated the effects of the antioxidant compound and major organosulfur garlic derivative, S-allylcysteine, on lipid peroxidation and mitochondrial dysfunction induced by 3-nitropropionic acid in synaptosomal fractions from rat brain. 3-Nitropropionic acid, at concentrations ranging 0.75-2.5 mM, produced enhanced levels of lipid peroxidation, while increasing concentrations of S-allylcysteine (0.1-2 mM) decreased the peroxidative action of 3-nitropropionic acid (1 mM) in synaptosomal fractions in a concentration-dependent manner. S-Allylcysteine (0.75 mM) also prevented the 3 nitropropionic acid (1mM)-induced mitochondrial dysfunction. These findings suggest that the protective actions that S-allylcysteine exert on the in vitro neurotoxicity induced by 3-nitropropionic acid are mediated by its antioxidant properties. PMID- 16377448 TI - Program factors related to women's substance abuse treatment retention and other outcomes: a review and critique. AB - This study examined program factors related to women's substance abuse treatment outcomes. Although substance abuse research is traditionally focused on men, some more recent studies target women. A systematic review of 35 empirical studies that included solely women subjects or that analyzed female subjects separately from male subjects revealed five elements related to women's substance abuse treatment effectiveness; these are (1) single- versus mixed-sex programs, (2) treatment intensity, (3) provision for child care, (4) case management and the "one-stop shopping" model, and (5) supportive staff plus the offering of individual counseling. Although all 35 studies contribute to the knowledge base, critiques of six areas of design weakness in the studies were included to provide directions for future studies; these are (1) lack of a randomized controlled design, (2) nondisentanglement of multiple conditions, (3) lack of a consistent definition for treatment factors and outcomes, (4) small sample size, (5) lack of thorough program description, and (6) lack of thorough statistical analyses. PMID- 16377447 TI - A comparative study of morphine treatment regimen prior to mating and during late pregnancy. AB - Pre-mating treatment of female rats with morphine may have long-term effects. In this study, we analyzed the effects of two types of morphine sulfate pre treatment: during pre-mating (5.0 mg/kg on alternate days for a total of seven doses) and during pregnancy (3.5 mg/(kgday) for 5 days starting on day 17 of pregnancy during early lactation. In order to evaluate possible morphine-induced behavioral changes, dams were tested for maternal behavior and locomotor activity during early lactation, and striatal and hypothalamic concentrations of dopamine and their metabolites and serum levels of corticosterone were measured. Maternal behavior was disrupted only in animals treated with morphine sulfate during pregnancy and challenged acutely (1.5 mg/kg) during lactation. Pre-mating treatment with morphine sulfate-induced changes in responses with increased locomotor activity, striatal dopamine turnover and serum corticosterone levels. None of these parameters were affected by morphine sulfate pre-treatment during late pregnancy. These data suggest that morphine has specific long-term and sometimes addictive-like effects on actively reproductive female animals that vary with the pre-treatment period, late pregnancy being particularly sensitive for effects on maternal behavior. PMID- 16377449 TI - An interactive voice response program to reduce drinking relapse: a feasibility study. AB - Substance-abusing patients often relapse soon after undergoing treatment, thus requiring intensive aftercare or re-treatment. More efficient monitoring and follow-up of patients could contribute to better treatment outcomes. This study evaluated the feasibility of a computer-automated interactive voice response (IVR) system to reduce relapse following discharge from residential treatment. Sixty participants completing a residential treatment program and meeting DSM-IV criteria for alcohol dependence were randomized to three groups: (1) daily IVR reporting with personal follow-up on noncompliant callers; (2) daily IVR reporting without follow-up; or (3) no IVR reporting (control group). At 30, 90, and 180 days after discharge, participants were interviewed to obtain timeline follow-back drinking data and completed the Work and Social Adjustment Scale, Obsessive-Compulsive Drinking Scale, SF-36, and Drinker Inventory of Consequences. This pilot study suggests that using automated IVR technology to monitor clients after discharge is feasible and warrants further research and development. IVR systems also provide the potential for delivering individualized feedback. PMID- 16377450 TI - Exposure to violence among substance-dependent pregnant women and their children. AB - This study examined the prevalence of exposure to violence among drug-dependent pregnant women attending a multidisciplinary perinatal substance abuse treatment program. Participants (N = 715) completed the Violence Exposure Questionnaire within 7 days after their admission to the program. Their rates of lifetime abuse ranged from 72.7% for physical abuse to 71.3% for emotional abuse to 44.5% for sexual abuse. Their rates of abuse remained high during their current pregnancy, ranging from 40.9% for emotional abuse to 20.0% for physical abuse to 7.1% for sexual abuse. Nearly one third of the women reported having physical fights with their current partner (lifetime), and 25% of these women reported that children were present during those physical fights. A total of 30% of the women perceived a need for counseling regarding exposure to violence for themselves and 15% perceived a need for counseling for their children. Study findings confirm previous reports of high rates of abuse and violence exposure among substance abusing pregnant women and their strong need for counseling for psychosocial sequelae. This study affirmed the value of routine screening for violence exposure in this at-risk population as well as the need to train therapists in specific strategies for helping such women address this complex array of problems. PMID- 16377451 TI - Outpatient substance abuse treatment and HIV prevention: an update. AB - Testing and counseling, along with community outreach, have been identified as valuable in the prevention of human immunodeficiency virus (HIV) and other blood borne diseases. This article assesses the extent to which outpatient substance abuse treatment (OSAT) programs provide such services. Longitudinal data for 1988 2000 were analyzed from the National Drug Abuse Treatment System Survey (NDATSS). Random-effects regression was used to examine factors associated with the provision of prevention services. HIV testing, which had became more common between 1990 and 1995, continued to proliferate between 1995 and 2000. The proportion of units that provide HIV testing and counseling increased from 66% to 86%. The proportion of units that provide HIV community outreach increased significantly before 1995 but then slightly decreased from 77% to 73% between 1995 and 2000. In conclusion, HIV testing and counseling widely proliferated in OSAT care. However, OSAT units remain less likely to offer HIV community outreach services. PMID- 16377452 TI - Contrasting faith-based and traditional substance abuse treatment programs. AB - This article (a) discusses the definition of faith-based substance abuse treatment programs, (b) juxtaposes Durkheim's theory regarding religion with treatment process model to highlight key dimensions of faith-based and traditional programs, and (c) presents results from a study of seven programs to identify key program dimensions and to identify differences/similarities between program types. Focus group/Concept Mapping techniques yielded a clear "spiritual activities, beliefs, and rituals" dimension, rated as significantly more important to faith-based programs. Faith-based program staff also rated "structure and discipline" as more important and "work readiness" as less important. No differences were found for "group activities/cohesion" and "role modeling/mentoring," "safe, supportive environment," and "traditional treatment modalities." Programs showed substantial similarities with regard to core social processes of treatment such as mentoring, role modeling, and social cohesion. Implications are considered for further research on treatment engagement, retention, and other outcomes. PMID- 16377453 TI - Heroin maintenance treatment for chronic heroin-dependent individuals: a Cochrane systematic review of effectiveness. AB - The provision of prescribed heroin to chronic heroin-dependent individuals failing other treatments has been supported during the last 70 years on the ground that the first goal of interventions on drug users is to keep them in treatment to protect them from criminal activities and to promote social integration. To assess heroin prescription effectiveness, we conducted a Cochrane systematic review of all relevant randomized controlled trials. We searched MEDLINE, EMBASE, CINAHL, and the Cochrane Library and contacted leading researchers for ongoing studies. We found 19 eligible studies, of which 4 met our inclusion criteria (577 patients). In 1 study, patients in the heroin arm remained in treatment longer than those in the methadone arm (n = 96, RR = 2.82, 95% CI = 1.70-4.68); in 2 studies, there was no difference; and in 1 study, patients given heroin left the study earlier than those given methadone (n = 235, RR = 0.79, 95% CI = 0.68-0.90). Heroin was more effective than methadone in refraining people from using street heroin in 2 studies (n = 96, RR = 1.10, 95% CI = 0.79-1.53; n = 51, RR = 0.33, 95% CI = 0.15-0.72). In 1 study, heroin reduced the risk of being charged (RR = 0.32, 95% CI = 0.14-0.78); 2 studies showed no difference, and another 2 studies adopted a multidomain outcome enclosing criminal offense and social functioning and found improvements with heroin + methadone over methadone only. It is unclear if heroin attracts people in treatment; those in treatment use less street heroin and are likely to have less criminal activities. This review systematizes and compares studies showing some inconsistencies between their aims, their adopted outcomes, and their conclusions drawn from results. PMID- 16377454 TI - Quality of care for substance use disorders in patients with serious mental illness. AB - We assessed the quality of care for substance use disorders (SUDs) among 8,083 patients diagnosed with serious mental illness from the VA mid-Atlantic region. Using data from the National Patient Care Database (2001-2002), we assessed the percentage of patients receiving a diagnosis of SUD, percentage beginning SUD treatment 14 days or earlier after diagnosis, and percentage receiving continued SUD care 30 days or less. Overall, 1,559 (19.3%) were diagnosed with an SUD. Of the 1,559, 966 (62.0%) initiated treatment and 847 (54.3%) received continued care. Although patients diagnosed with bipolar disorder were more likely to receive a diagnosis of SUD than those diagnosed with schizophrenia or schizoaffective disorder (22.7%, 18.9%, and 17.7%, respectively; chi(2) = 26.02, df = 2, p < .001), they were less likely to initiate (49.1%, 70.7%, and 68.6%, respectively; chi(2) = 59.29, df = 2, p < .001) or continue treatment (39.9%, 63.2%, and 62.2%, respectively; chi(2) = 72.25, df = 2, p <. 001). Greater efforts are needed to diagnose and treat SUDs in patients with serious mental illness, particularly for those with bipolar disorder. PMID- 16377455 TI - Integrated versus parallel treatment of co-occurring psychiatric and substance use disorders. AB - The study examines 1-year treatment outcomes of 216 individuals with co-occurring severe and persistent mental illness and substance use disorders who were assigned to an integrated or parallel treatment condition. Comparisons indicated that the integrated group achieved greater reductions in the incidence of psychiatric hospitalization and arrest. The results of this study support the enhanced effectiveness of integrated treatment in decreasing the use of higher cost crisis-oriented services in clients with severe mental illness and substance use disorders. PMID- 16377456 TI - Preliminary evidence that prenatal alcohol damage may be visible in averaged ultrasound images of the neonatal human corpus callosum. AB - Brain damage consequent to prenatal alcohol exposure can be detected by measurements of the corpus callosum in the midline magnetic resonance (MR) brain image in adolescents and adults. The present article extends this finding into the neonatal period, when the power of detection to ameliorate the quality of the child's future life is greatest. The midline corpus callosum of the very young infant can be located reliably in multiple frames of clinical transfontanelle ultrasound. We studied a sample of 18 children aged 17 weeks or less, 7 of whom were exposed to high levels of alcohol prenatally and 11 of whom were not exposed or only minimally exposed. The midline callosum of each child was imaged up to 50 times by a standard clinical device, and coplanar subsets of these series were averaged with reference to fiducial image structures. On each average image four semilandmark points were set and their configuration quantified by standard landmark methods. The angle between the terminal bulb of splenium and the long axis of the callosal outline classifies four of the seven exposed infants as different from all 11 of the unexposed infants. This simple angle measurement upon averaged ultrasound images of the human neonatal midline corpus callosum, perhaps a version of the long-sought "biomarker of prenatal alcohol damage," may be able to discriminate baby brains affected by prenatal alcohol exposure from those that were unaffected. PMID- 16377457 TI - Baclofen-induced reduction of alcohol reinforcement in alcohol-preferring rats. AB - Recent studies have demonstrated that treatment with the gamma-aminobutyric acid (GABA(B)) receptor agonist, baclofen, reduces alcohol intake in selectively bred Sardinian alcohol-preferring rats tested under the homecage two-bottle "alcohol versus water" choice regimen. This study was designed to investigate whether baclofen also reduces alcohol-reinforcing effects in Sardinian alcohol-preferring rats. To this aim, sP rats were trained to lever press for oral alcohol (15%, vol/vol) or sucrose (0.3%, wt/vol; included as alternative reinforcer to evaluate the specificity of baclofen effect on alcohol reinforcement) under a fixed ratio schedule of 4. Once steady levels of alcohol or sucrose self-administration behavior were established, the effects of acutely administered baclofen (0, 1.7, and 3 mg/kg, intraperitoneal [ip]) and naloxone (0, 1, and 3 mg/kg, ip; included as reference compound) on alcohol- or sucrose-reinforced responding were evaluated. Baclofen administration dose dependently, although not specifically, reduced alcohol-reinforced responding to an extent comparable to that of naloxone. Baclofen also produced a dose-dependent and specific delay in the onset of alcohol-reinforced responding, suggesting that it suppressed the rats' motivation to start drinking alcohol. These data are discussed in terms of adding further support to the hypothesized involvement of the GABA(B) receptor in the neural system mediating alcohol reinforcement. These data are also in agreement with the results of recent preliminary clinical studies suggesting that baclofen may have therapeutic efficacy in the treatment of alcohol dependence. PMID- 16377458 TI - Effect of natural and synthetic polyamines on ethanol intake in UChB drinker rats. AB - Because of the important glutamatergic mediation of the behavioral effects of ethanol, glutamatergic agents have attracted attention for the treatment of ethanol abuse and dependence in preclinical and clinical studies. In the present study, we investigated the effect of pharmacological doses of the natural polyamines putrescine, spermine, and spermidine and the synthetic polyamine N,N' bis-(3-aminopropyl)cyclohexane-1,4-diamine (DCD) on alcohol consumption in a free choice paradigm carried out in genetically high-ethanol-consumer UChB rats. Short 3-day treatment with either polyamine, administered p.o., significantly reduced ethanol intake without modifying water and food intakes. Neither polyamine was able to increase markedly blood acetaldehyde in rats submitted to a standard challenge dose of ethanol, to rule out a disulfiram-like effect. Besides, blood ethanol disappearance after a test dose of ethanol was not affected by the synthetic polyamine DCD. Long-term treatment with DCD dose-dependently reduced ethanol intake in UChB rats without producing any observable effect on overt behavior, food consumption, and total fluid intake. The present results indicate that pharmacological doses of polyamines can reduce ethanol consumption in genetically drinking rats without producing significant side effects, suggesting that modulation of brain N-methyl-d-aspartate receptors by polyamines could represent a suitable strategy to reduce appetite for ethanol. However, caution must be exercised in interpreting the results because polyamines can also affect neuronal excitability by acting at other receptor targets, such as AMPA and kainate receptors, as well as at some voltage-dependent ion channels. PMID- 16377460 TI - Development of an ethanol model using social insects: V. Honeybee foraging decisions under the influence of alcohol. AB - We examined the effect of ethanol on a complex decision process in honeybees. In many contexts (e.g., flower patches containing both blue and white flowers), honeybees have been shown to make decisions concerning which flowers to visit based on the reward's caloric value offered by competing flower types. However, under some contexts (e.g., flower patches containing both blue and yellow flowers) honeybees ignore the energetic differences that exist between rewards. When this occurs, some bees show extreme fidelity to one flower type, and other bees simultaneously show extreme loyalty to the competing flower type. In this set of experiments we use different combinations of flower colors to elicit these different context-specific behaviors, and examine response under the influence of ethanol in conditions with rewards differing in either quality or quantity. Alcohol affected the same decision situation differently in the alternative contexts we gave foragers. In the blue-white dimorphic flower patch context, foragers in the alcohol condition no longer followed the species' typical behavior of selectively choosing the higher energetic reward; they chose flowers randomly as if rewards did not differ between flower colors. However, in the blue yellow dimorphic flower patch context, foragers in the alcohol condition continued their species-typical behavior of favoring their initial flower-color choice. That is, alcohol had no effect on behavior in the latter context. This pattern of context-dependent effects of alcohol was not associated with motor impairment in either context because flower visitation rate increased when bees were given ethanol. The brain regions responsible for alternative context behaviors (using the Drosophila model), or at least receptors within the same region responsible for these context-dependent behaviors, have very different sensitivities to alcohol, which suggests that differing neural processes are involved. PMID- 16377459 TI - The neuropeptide-Y Y5 receptor antagonist L-152,804 decreases alcohol self administration in inbred alcohol-preferring (iP) rats. AB - Neuropeptide-Y (NPY) is the most abundant and widely distributed peptide in the mammalian central nervous system and increases feeding behavior through actions at the Y5 receptor subtype. Recent pharmacological evidence indicates that NPY activity at this receptor subtype can modulate ethanol reinforcement. The purpose of this study was to determine if NPY Y5 receptor antagonism reduces ethanol self administration and reinforcement in a rodent genetic animal model of alcoholism. Selectively inbred alcohol-preferring (iP) rats were trained to voluntarily consume ethanol (10% vol/vol) versus H2O in a 24-h two-bottle choice test. An additional group of iP rats was trained in operant ethanol self-administration to lever press on a fixed-ratio 1 schedule for ethanol (10% vol/vol) reinforcement. Following establishment of baseline intake or ethanol-reinforced responding, iP rats were injected with L-152,804 (0-20 mg/kg) prior to two-bottle or operant ethanol self-administration sessions. In the two-bottle choice test, L-152,804 (3 and 10 mg/kg, ip) significantly reduced ethanol intake (g/kg) at 4- and 6-h postinjection and had no effect on food intake. In the operant procedure, L 152,804 (10 and 20 mg/kg, ip) significantly reduced both the dosage of self administered ethanol (g/kg/1-h) and the total number of ethanol-reinforced responses. No effect was observed on latency to the first response or the number of inactive lever presses. These results indicate that blockade of NPY Y5 receptor activity decreases both voluntary ethanol drinking and ethanol reinforcement in a rodent genetic animal model of alcoholism. For this reason, NPY Y5 receptor antagonists may be useful in medical management of alcohol abuse and alcoholism in the human population. PMID- 16377461 TI - Effects of alcohol on thermoregulation during mild heat exposure in humans. AB - We investigated the effects of alcohol on thermoregulatory responses and thermal sensations during mild heat exposure in humans. Eight healthy men participated in this study. Experiments were conducted twice for each subject at a room temperature of 33 degrees C. After a 30-min resting period, the subject drank either 15% alcohol (alcohol session) at a dose of 0.36 g/kg body weight or equal volume of water (control session). Skin blood flow and chest sweat rate in the alcohol session significantly increased over those in controls 10 min after drinking. Deep body temperature in the alcohol session started to decrease 20 min after the onset of sweating and eventually fell 0.3 degrees C lower than in the controls. Whole body hot sensation transiently increased after alcohol drinking, whereas it changed little after water drinking. The increased "hot" sensation would presumably cause cool-seeking behavior, if permitted. Thus, alcohol influences thermoregulation so that body core temperature is lowered not only by automatic mechanisms (sweating and skin vasodilation) but also behaviorally. These results suggest that decreases in body temperature after alcohol drinking are not secondary to skin vasodilation, a well-known effect of alcohol, but rather result from a decrease in the regulated body temperature evidenced by the coordinated modulation of various effectors of thermoregulation and sensation. PMID- 16377462 TI - Inhibition of bone repair in a rat model for chronic and excessive alcohol consumption. AB - Alcohol abuse is associated with increases in both the incidence of fractures and complications in fracture healing. The purpose of this study was to determine the dose-dependent effects of ethanol on bone repair in a rat model. Three-month-old male Wistar rats were continuously fed liquid diets containing ethanol as either 36% or 26% of total calories or control diets for 6 weeks. Then, a bone repair model was created in all rats. Bone healing and liver metabolism were evaluated 7 weeks after bone injury. For each dose, there were three ethanol-feeding groups receiving (1) ethanol for 13 weeks, (2) control diet for 13 weeks (pair-fed), and (3) ethanol before bone injury and control diet (pair-fed) after injury. Another group was fed ethanol (36%) before injury and given control diet ad libitum after injury. There were also two nutritional controls consuming control diet and standard rat chow ad libitum for 13 weeks. Abnormal liver metabolism was evident at the higher ethanol dose - increases in cytochrome P4502E1 specific activity (5 fold; P < .01), triglyceride content (4-fold; P < .02), and liver weight (25%; P = .05) - compared with pair-fed controls. The higher dose of ethanol resulted in deficient bone repair when compared with rats receiving ethanol-free control diet by pair-feeding: 26% less (P = .02) rigidity of the repaired bone, 41% less (P = .02) intrinsic stiffness, 24% less intrinsic strength (P = .05), and 14% less (P = .001) ash density of the repair tissue. The reduced food consumption of ethanol fed rats compared with that in the nutritional controls did not contribute to this deficiency. Furthermore, removal of ethanol (as 36% of calories) from the diet after bone injury completely restored normal bone healing and nearly normalized the liver metabolism. The lower ethanol dose (26% of calories) had a minimal effect on liver metabolism and bone repair. We conclude that ethanol (as 36% of calories) in the rat diet, especially during the postinjury period, was solely responsible for the observed inhibition of bone repair. PMID- 16377464 TI - Heparins, low-molecular-weight heparins, and pentasaccharides. AB - Elderly patients require special consideration when administered anticoagulants because of age-related alterations in renal function, protein binding, and increased bleeding risk. Unfractionated heparin can be used in most patients but difficulties with dosing and monitoring often lead to inadequate anticoagulation. Low-molecular-weight heparin has more predictable pharmacokinetics than conventional heparin, but requires dose adjustments in renal impairment and obesity. Fondaparinux is a synthetic pentasaccharide that is being used increasingly for both treatment and prophylaxis of venous thromboembolism. The immune-mediated form of heparin-induced thrombocytopenia is a syndrome with thrombocytopenia or thrombosis in the setting of heparin use. Heparin-induced thrombocytopenia must be identified early, and treated with argatroban or lepirudin to avoid life-threatening complications. PMID- 16377465 TI - Warfarin pharmacology, clinical management, and evaluation of hemorrhagic risk for the elderly. AB - Elderly patients as a group may present more of a challenge in managing warfarin therapy because of alterations in pharmacokinetics from other medications, diet, and disease; pharmacodynamic changes; increased risk for hemorrhage; and difficulty in monitoring. The elderly, however, may derive the most benefit from warfarin therapy for certain indications, such as the prevention of stroke in atrial fibrillation or recurrent events following deep venous thrombosis. Warfarin can be managed as effectively as in other populations with careful attention to these issues. PMID- 16377466 TI - New anticoagulant agents: direct thrombin inhibitors. AB - Decades of research have been devoted to developing effective, safe, and convenient anticoagulant agents. In recent years, much emphasis has been placed on the development of direct thrombin inhibitors (DTIs) that offer benefits over agents like heparin and warfarin including the inhibition of both circulating and clot-bound thrombin; a more predictable anticoagulant response, because they do not bind to plasma proteins and are not neutralized by platelet factor 4; lack of required cofactors, such as antithrombin or heparin cofactor II; inhibiting thrombin-induced platelet aggregation; and absence of induction of immune mediated thrombocytopenia. Various injectable DTIs are currently available and used for many indications. In addition, research is now focusing on oral DTIs that seem promising and offer various advantages, such as oral administration, predictable pharmacokinetics and pharmacodynamics, a broad therapeutic window, no routine monitoring, no significant drug interactions, and fixed-dose administration. PMID- 16377467 TI - Antiplatelet agents and arterial thrombosis. AB - There is an increase in arterial thrombotic events in the elderly. Elderly patients are more likely to have associated diseases, such as diabetes, hypertension and hypercholesterolemia, and when age is confounded by these other predisposing factors, the risk of an arterial ischemic event increases disproportionately. Antithrombotic therapy for geriatric patients is underused, even when one adjusts for potential drug contraindications. This article focuses on the action of the currently available antiplatelet agents--aspirin, clopidogrel, and glycoprotein IIb/IIIa (GPIIb/IIIa) receptor antagonists, and assesses their effects in different disease states, with special attention to data that examine the geriatric population. PMID- 16377468 TI - Pathophysiology of venous thrombosis, thrombophilia, and the diagnosis of deep vein thrombosis-pulmonary embolism in the elderly. AB - This article reviews Virchow's triad to provide clinicians with the pathophysiologic basis for the development of venous thromboembolism (VTE). Armed with these data physicians should be vigilant of the development of VTE when these factors are present. The second clinical point is the appropriate need to evaluate elderly patients for primary or secondary thrombophilia. By using statistical probability and clinical history selected geriatric patients with VTE should be assessed for the etiology of their hypercoagulable state. Finally, evaluating symptomatic patients for VTE should be structured to use the appropriate testing to confirm the diagnosis. PMID- 16377469 TI - Prevention of venous thromboembolism in the geriatric patient. AB - Elderly patients immobilized because of an acute medical illness or surgery have a very high risk of developing venous thromboembolism (VTE). Aggressive pharmacologic prophylaxis is necessary and should be initiated either at admission for a medical condition or shortly after surgery. Aggressive prophylaxis may result in fewer patients developing VTE in the hospital and ultimately lead to fewer patients requiring full-dose anticoagulation for VTE. Mechanical prophylaxis can be used as an adjunct to an anticoagulant-based regimen but should only be used as primary prophylaxis when there is a contraindication, such as active bleeding. It is recommended that the clinician carefully evaluate the elderly patient's creatinine clearance and weight before prescribing anticoagulants, particularly when using fixed dosing regimens. PMID- 16377470 TI - Anticoagulant treatment of deep vein thrombosis and pulmonary embolism. AB - Venous thrombosis is a common disease. As the mean age of the population increases, so does the incidence of venous thromboembolism. Anticoagulant therapy is equally effective in young and older patients, and can reduce substantially the associated morbidity and mortality. When considering long-term oral anticoagulant therapy in older patients, however, careful ongoing evaluation is imperative to ensure that the risk of bleeding does not outweigh the antithrombotic benefits. PMID- 16377471 TI - Antithrombotic and thrombolytic therapy for ischemic stroke. AB - Thrombolytic and antithrombotic agents form the cornerstone of stroke treatment and prevention. Recombinant tissue plasminogen activator improves outcome in patients treated within 3 hours of stroke onset. Emerging trials are directed to extend the therapeutic window and identify agents that could provide better safety profiles. Large, randomized trials have also highlighted the effectiveness and safety of early and continuous antiplatelet therapy in reducing atherothrombotic stroke recurrence. Aspirin has become the antiplatelet treatment standard against which several other antiplatelet agents have been shown to be more effective. The prevention of cardioembolic stroke is best accomplished with oral anticoagulation, barring any contraindications. PMID- 16377472 TI - Antithrombotic therapy in atrial fibrillation. AB - Warfarin is highly effective at reducing the risk of stroke in atrial fibrillation. The benefit of oral anticoagulant therapy strongly outweighs the risk in most patients with atrial fibrillation. More data are needed to define better the overall risk-to-benefit ratio for patients age 80 years and greater. Because a significant proportion of elderly individuals may not be optimal candidates for anticoagulant therapy, alternative stroke prevention strategies must continue to be evaluated while redoubling efforts to understand the mechanisms underlying atrial fibrillation and thrombogenesis. PMID- 16377473 TI - Chronic antithrombotic therapy in post-myocardial infarction patients. AB - Recent advances in the acute treatment of myocardial infarction (MI) have allowed more to survive the initial event in coronary artery disease. Increasing importance has been placed on secondary-prevention strategies, with more attention directed toward an optimal therapeutic regimen for post-MI patients. When it comes to the subpopulation of the elderly who have had an MI, however, current practice relies less on population-specific evidence and more on extrapolation of current data. This article reviews the pathogenesis of coronary heart disease and MI, and highlights recent major trials that have explored antiplatelet and anticoagulant medications in the chronic treatment of the post MI population. Special consideration is given to the geriatric community and its unique challenge regarding chronic antithrombotic therapy. PMID- 16377474 TI - Antithrombotic therapy in peripheral arterial disease. AB - The management of elderly patients with peripheral arterial disease requires a multidisciplinary and individualized approach, especially for patients requiring intervention and for those on antithrombotic therapy. Communication between the patient's primary physician, consulting medical specialists, and vascular surgeon is essential because all may contribute synergistically to deliver optimal care to the patient. This article reviews the pathophysiology of peripheral arterial disease, and data regarding the use of antiplatelet and anticoagulant agents. PMID- 16377475 TI - Perioperative management of oral anticoagulation. AB - Based on an individual assessment of risk factors for arterial or venous thrombosis and the risk of postoperative bleeding, this article outlines the preoperative and postoperative approach to anticoagulant management. Preceding this is a brief description of the therapies most commonly used in the perioperative period. The prevention of arterial thromboembolism is considered separately from the prevention of venous thrombosis. PMID- 16377476 TI - Transcranial color Doppler flow imaging in detecting severe stenosis of the intracranial vertebral artery: a prospective study. AB - OBJECTIVE: The aims of this prospective study were to investigate the manifestations of the hemodynamics and morphology of a normal intracranial vertebral artery (VA) and diffusely severe or localized critical vertebral artery stenosis (VAS) on transcranial color Doppler flow imaging (TCDFI) and to discuss the criteria of TCDFI in detecting severe VAS. METHODS: There were 30 patients suspected of having an intracranial VAS in a patient group and 30 healthy volunteers in a control group. All the patients underwent both TCDFI and digital subtraction angiography (DSA). The intracranial VA was imaged through the suboccipital window by TCDFI. The size and tortuousness of the vessels were observed. The hemodynamics of those vessels were measured by means of peak systolic velocity (PSV) and pulsatility index (PI). RESULTS: In the control group, the VA junction with basilar artery appeared as Y shaped and filled with well-distributed blue coded on color imaging, which showed flow below the baseline with fast early systolic acceleration, and normal PSV ranged from 0.42 to 0.80 m/s on pulse wave Doppler. In the patients with diffusely severe VAS (>70% lumen reduction, the length of plaque >1 cm or multiple plaques) or localized critical VAS (>90% lumen reduction) diagnosed by TCDFI and confirmed with DSA, a narrowed and tortuous VA was demonstrated with minimal flow on color Doppler imaging and decreased 50% of normal PSV and PSV <0.30 m/s accompanied by slow early systolic acceleration on Doppler spectrum. There was a significant difference (P<.05 or P<.01) in the hemodynamic parameters (PSV and PI) between patients with severe VAS and healthy subjects. CONCLUSION: TCDFI is a useful noninvasive tool in assessing the hemodynamics of intracranial arteries. The sensitivity was 90.9% and the specificity was 75.0% by using PSV <0.30 m/s and 50% reduction of normal PSV as low cutoff criteria in the diagnosis of diffusely severe or localized critical VAS with TCDFI. PMID- 16377477 TI - Role of fine needle aspiration and tru cut biopsy in diagnosis of mucinous carcinoma of breast--from a radiologist's perspective. AB - This study was to assess whether fine needle aspiration (FNA) or core biopsy would allow diagnosis of mucinous carcinoma of breast. In 37 mucinous carcinoma in 34 patients, 20 lesions had FNA and 24 lesions with core biopsy. FNA achieved a sensitivity of 66.7% in diagnosis of malignant lesions and 56% sensitivity in diagnosis of mucinous carcinoma. Core biopsy achieved 100% sensitivity and accuracy in the diagnosis of malignant lesions and mucinous carcinoma. PMID- 16377478 TI - CT screening for lung cancer: significance of diagnoses in its baseline cycle. AB - PURPOSE: The aim of this study was to assess the significance of Stage I diagnoses of lung cancer in the baseline cycle of screening for this disease, with special reference to the potential for overdiagnosis. METHODS: We reviewed all 69 cases of Stage I lung cancer diagnosis resulting from our baseline CT screening. Among these 69 cases of lung cancer, 24 presented as solid, 30 as part solid, and 15 as nonsolid nodules. The extent to which these represented genuine malignancy was assessed by a panel of experts on lung pathology, and the "aggressiveness" of these cases was addressed by the criterion of the tumor's volume doubling time being less than 400 days. RESULTS: The expert panel confirmed all 69 cases as representing genuine malignancy. Among the 69 cases without evidence of metastases, the proportion that satisfied the aggressiveness criterion was 60/69=87%. The corresponding proportions by presentation as solid, part-solid, and nonsolid nodule were 23/24 (96%), 27/30 (90%), and 10/15 (67%), respectively. CONCLUSIONS: In baseline CT screening for lung cancer, overdiagnosis of the disease is uncommon, with cases presenting as a nonsolid nodule a possible exception to this. PMID- 16377479 TI - Twenty-four-hour thallium-201 late redistribution imaging enhances the detection of myocardial viability after myocardial infarction. AB - BACKGROUND: Thallium-201 (201Tl) myocardial perfusion imaging has been widely used for evaluation of myocardial ischemia/viability after myocardial infarction. The 3- to 4-h early redistribution imaging has underestimated a considerable part of viable myocardium, while the 24-h late redistribution imaging may enhance the detection of myocardial ischemia/viability, but remains controversial. METHODS: Thirty-eighty patients with myocardial infarction underwent the initial, 3-h, and 24-h redistribution imaging after intravenous injection of 148-185 MBq 201Tl. Image quality analysis was performed using a four-grade model: excellent, good, moderate, and poor. The initial and 3-h images, the initial and 24-h images, and the 3- and 24-h images were compared double-blinded. RESULTS: The 3- and 24-h images showed no significant differences in image quality according to the four grade model (P=.3580). Out of the 194 abnormal segments based on the initial imaging, 60 (31%) segments improved by at least one grade on the 3-h imaging, while 86 (44%) segments improved by at least one grade on the 24-h imaging. The 24-h late imaging detected more viable myocardium than the 3-h imaging did, with a significant difference (chi2=7.4235, P=.0064). There were 164 abnormal segments on the 3-h imaging, with an average 30% (48) segments improved by at least one grade on the 24-h imaging. There were 134 initial abnormal segments without any improvement on the 3-h imaging. Out of these segments, the 24-h late redistribution imaging detected additional redistribution in 30 segments, taking up 22%. The mean global score on the 3-h imaging significantly decreased compared to that on the initial imaging (t=5.71, P<.0001), and the mean global score on the 24-h imaging further decreased significantly compared to that on the 3-h imaging (t=6.28, P<.0001). CONCLUSIONS: Twenty-four-hour late 201Tl imaging, with satisfactory image quality, enhances the detection of myocardial viability after myocardial infarction. PMID- 16377480 TI - Aneurysm shrinkage after endovascular repair of aortic diseases. AB - BACKGROUND: There are two graft materials for endovascular repair of aortic diseases, i.e., polyester and expanded polytetrafluoroethylene (ePTFE). The latest reports have suggested that there is graft-specific difference in outcomes. The purpose of this article was to evaluate the difference in terms of aneurysm shrinkage. PATIENTS AND METHODS: Eighty-six patients who underwent endovascular repair of aortic diseases were included. Forty patients had true aortic aneurysms, 8 had aortic pseudoaneurysms, and 38 had aortic dissections. Eighteen patients with true aortic aneurysms were treated with stent grafts fabricated with polyester, while the other 68 patients, including 22 patients with true aneurysms, 8 patients with pseudoaneurysms, and 38 patients with aortic dissections, were treated with stent grafts fabricated with ePTFE. All patients were followed-up by computed tomography (CT) for more than 1 year. The mean follow-up term was 28 months. Computed tomography confirmed that there were sufficiently long necks, and the aneurysm or the entry tear was completely excluded without any endoleak in all patients. The diameter of the preoperative lesion was compared with that measured on follow-up CT at 1 year and at the end of the follow-up term. Increase or decrease in the diameter by more than 5 mm was defined as a significant diameter change. RESULTS: Aneurysm shrinkage was observed in 40% of patients with true aneurysms, 88% of patients with pseudoaneurysms, and 55% of patients with aortic dissections at 1 year. There was no significant increase in patients with aneurysm shrinkage at the end of follow up in any groups. In the case of true aortic aneurysms, shrinkage of aneurysms was observed more frequently with polyester-fabricated stent grafts (67%, 13/18) than with ePTFE-fabricated ones (18%, 4/22) at 1 year (P<.01). In contrast, expansion of aneurysms was observed only in patients treated with ePTFE (14%, 3/22). Shrinkage of the descending aorta was observed in 55% of patients with acute aortic dissections and 36% of patients with chronic aortic dissections. There was no case with aortic enlargement in either group. There was no significant difference between acute and chronic dissection in terms of shrinkage of the descending aorta. CONCLUSION: Expanded polytetrafluoroethylene appears to be effective for the treatment of pseudoaneurysms and aortic dissections. However, polyester seems to be more effective than ePTFE when true aneurysms are to be treated. PMID- 16377481 TI - Bowel wall enhancement in magnetic resonance colonography for assessing activity in Crohn's disease. AB - BACKGROUND: This study focuses on the correlation of the signal intensity (SI) of the bowel wall in magnetic resonance (MR) colonography with the inflammation activity in Crohn's disease. MATERIAL AND METHODS: We performed MR colonography in 42 patients and compared the change of the SI of the bowel wall (T1 weighted/paramagnetic contrast medium) with colonoscopic findings. RESULTS: Change of the SI and colonoscopically assessed inflammatory activity are significantly correlated (r=.676, P85% of patients. A nasogastric tube was left in situ postoperatively in 40% versus 66% of patients in the United States and Europe, respectively, and was removed about 3 days postoperatively. It took 3 to 4 days until 50% of the patients first tolerated liquids and 4 to 5 days until 50% of patients were eating and having a bowel movement. Postoperative ileus was found to persist for over 5 days in approximately 45% of patients. Mean length of postoperative hospital stay was over 10 days in the United Kingdom, France, Germany, Italy, and Spain, and 7 days in the United States, compared with 2 to 5 days reported in trials of fast-track colonic surgery programs. CONCLUSIONS: Strategies that can contribute to improved recovery and reduced complications after colonic operations do not appear to be applied optimally in clinical practice across Europe and the United States. These findings indicate a potential for major improvements in outcomes and reduction of costs if peri- and postoperative care can be adjusted to be in line with published evidence. PMID- 16377497 TI - Colon and rectal complications after heart and lung transplantation. AB - BACKGROUND: Gastrointestinal complications of solid organ transplantation have been well described, but little attention has been paid to colorectal disorders in particular. The purpose of this study was to identify the incidence and severity of colorectal complications among a large cohort of heart and lung transplant recipients. STUDY DESIGN: We reviewed the medical records of heart, lung, and heart-lung transplant recipients at a single institution between 1978 and 2004. Complications were identified based on need for consultation, endoscopy, or operation by a colorectal surgeon after transplantation. RESULTS: Of 1,012 patients who received transplantations (530 heart, 435 lung, 47 heart lung), 56 patients (6%) required evaluation for 84 colorectal problems. Incidence of complications was 7% in lung transplant recipients, 6% in heart-lung transplant recipients, and 4% in heart transplant recipients. Forty-four events (52%) were considered major (diverticulitis, perforation, malignancy, and other) and 40 (48%) were minor (polyps, pseudo-obstruction treated medically or endoscopically, benign anorectal disease, and other). Twenty-three (27%) required colectomy and 9 (10%) necessitated anal operation. Thirty-six (43%) required less invasive interventions (endoscopy, minor anorectal procedures, and other). Eighteen (21%) were treated with medical therapy alone. Six patients died from colorectal disease (7%). CONCLUSIONS: Colorectal complications are a considerable source of morbidity and mortality after heart and lung transplantation. These complications occur more frequently in patients who undergo lung and heart-lung transplantation as compared with heart transplantation alone. PMID- 16377498 TI - Beta test results of a new system assessing competence in laparoscopic surgery. AB - BACKGROUND: There is currently a need for objective measures of surgical competence. Such measures should assess knowledge, judgment, and manual skills. The Fundamentals of Laparoscopic Surgery (FLS) program was developed by the Society of American Gastrointestinal and Endoscopic Surgeons to meet these criteria. The FLS assessment includes a multiple-choice cognitive test and a manual skills test. We present the results of validation studies of this novel assessment tool. STUDY DESIGN: Beta testing of the FLS examination was undertaken at 7 sites by 70 surgeons representing 4 levels of experience and training. Surgeons provided information about their prior experience and indicated a self assessment of their laparoscopic competence. Results were assessed by ANOVA followed by orthogonal contrasts. RESULTS: Cognitive performance by training level: There was no difference between fellows and staff in percentage of questions answered correctly, but there was a discrepancy between junior and senior residents and between residents and senior surgeons (p < 0.01). Cognitive performance by laparoscopic experience quartiles: There were notable contrasts between the first and second quartiles of experience (p < 0.02) and between the third and fourth quartiles (p < 0.01). No marked difference was found between the second and third quartiles. Cognitive performance compared with self-assessment: Test results were substantially different (p < 0.01) between test-takers who assessed themselves as "better than average" and those who assessed themselves as "average" or "below average." Manual skills performance by training level: The major difference was found between junior residents versus senior residents, fellows or staff (p < 0.01). Manual skills performance by laparoscopic experience level: Differences were primarily seen between the first two quartiles and the last two quartiles of laparoscopic experience (p < 0.001). Manual skills performance compared with self-assessment: Those who assessed themselves as "above average" in laparoscopic skill performed markedly better than those indicating they had "average" or "below average" skill (p < 0.01). CONCLUSIONS: Beta test results for the FLS examination demonstrate satisfactory reliability, appropriate psychometric properties, and substantial initial validity. The FLS project is one of the first validated surgical education efforts to assess the competence of surgeons in a specific field. PMID- 16377499 TI - Assessment of obesity-related comorbidities: a novel scheme for evaluating bariatric surgical patients. AB - BACKGROUND: Bariatric surgery serves as the superior means of achieving sustained weight loss and improvement in obesity-related comorbidities. Results of bariatric surgery have been reported qualitatively without standardized measurement of comorbidity response. The objective of this work was to develop a clinically based, standardized system for scaled assessment of the major comorbidities of obesity in patients undergoing bariatric surgery. STUDY DESIGN: We constructed a standardized grading scheme for the major comorbidities of obesity, with each condition scored from 0 to 5, according to severity. Data were prospectively collected on 226 patients. Ninety patients have already undergone gastric bypass and are being followed at regular intervals postoperatively. Longest current followup interval is 1 year. RESULTS: Preoperative evaluation of comorbidities identified a total of 1,356 medical disorders. Anatomic comorbidities were most prevalent as a category, although psychosocial impairment was the most common single condition. The majority of comorbidities in our patient population were graded mild (score of 1) to moderate (score of 3). Immediate (2 weeks) followup was available for all operated patients and ranged in number to 1 year postoperatively, depending on the date of operation. Statistically significant reduction in the severity of several comorbidities was observed at postoperative evaluation (p < 0.05). CONCLUSIONS: This scheme for assessment of obesity-related comorbidities facilitates evaluation of bariatric surgical patients. The system allows standardized preoperative characterization of a bariatric patient population and uniform postoperative longitudinal assessment of changes in comorbidities after weight reduction operation. PMID- 16377500 TI - Comparative analysis of vagotomy and drainage versus vagotomy and resection procedures for bleeding peptic ulcer disease: results of 907 patients from the Department of Veterans Affairs National Surgical Quality Improvement Program database. AB - BACKGROUND: The purpose of this study was to determine postoperative outcomes and risk factors for morbidity and mortality in patients requiring surgery for bleeding peptic ulcer disease (PUD). Vagotomy and drainage procedures are technically simpler but are usually associated with higher ulcer recurrence rates. In contrast, vagotomy and resection approaches offer lower ulcer recurrences but represent much more challenging operations and are associated with considerable morbidity and mortality. STUDY DESIGN: Data collected through the Department of Veterans Affairs National Surgical Quality Improvement Program database from 1991 to 2001 were submitted for stepwise logistic regression analysis for prediction of 30-day postoperative morbidity and mortality, rebleeding, and postoperative length of stay. The study population included all patients operated on for bleeding PUD within an 11-year period. RESULTS: The 30 day morbidity, mortality, and rebleeding rates were comparable between surgical groups. Age, American Society of Anesthesiologists class, presence of ascites, coma, diabetes, functional status, hemiplegia, and history of steroid use were predictors of postoperative death. Risk factors for rebleeding included dependent functional status, history of congestive heart failure, smoking, steroid use, and preoperative transfusions. Having a resective procedure, American Society of Anesthesiologists class, hemiplegia, history of COPD, and requiring ventilator assisted respirations before surgery were positively associated with increased length of hospital stay. CONCLUSIONS: No differences were observed in 30-day mortality, morbidity, or rebleeding rates between surgical groups. Having a resective procedure was a predictor of prolonged postoperative stay. Dependent status and chronic use of steroids were predictors of both rebleeding and postoperative mortality. PMID- 16377501 TI - Renal function is well maintained after use of left renal vein graft for vascular reconstruction in hepatobiliary-pancreatic surgery. AB - BACKGROUND: Advanced hepatobiliary-pancreatic malignancy occasionally involves major vasculatures, such as the portal vein or the inferior vena cava, and complete removal of the tumor is required for longterm survival. We used a left renal vein graft to reconstruct resected vessels in some patients. In this study, we evaluated early and late renal complications of this procedure. STUDY DESIGN: We identified 14 patients undergoing vascular reconstruction with use of a left renal vein graft in hepatobiliary-pancreatic surgery. Renal function and graft patency were assessed by observing serum creatinine levels and radiologic findings during perioperative and followup periods. RESULTS: Of these 14 patients, 7 were men and 7 were women. Diseases included hilar cholangiocarcinoma in two, gallbladder carcinoma in two, intrahepatic cholangiocarcinoma in one, pancreas carcinoma in five, hepatic metastasis in three, and mass-forming pancreatitis in one. No significant postoperative renal dysfunction was recognized, and the mean value of the maximal serum creatinine was 1.0 mg/dL during the perioperative period. Renal scintigraphy was performed in six patients postoperatively, and there was no significant left renal dysfunction. Mean followup time was 18 months after operation, and no severe renal dysfunction was found. Graft patency, which was assessed with enhanced abdominal CT, was well maintained after operation. CONCLUSIONS: The left renal vein could be safely obtained and used for vascular reconstruction in resection of hepatobiliary pancreatic malignancy without adverse effects on early and longterm renal function. PMID- 16377502 TI - Effect of bile contamination on immediate outcomes after pancreaticoduodenectomy for tumor. AB - BACKGROUND: The influence of preoperative biliary drainage on the postoperative course of patients undergoing pancreaticoduodenectomy (PD) remains controversial. Among drawbacks of biliary drainage, bile contamination and its consequences are incompletely evaluated. This study aimed to compare outcomes after PD in patients with sterile and those with infected bile. STUDY DESIGN: Seventy-nine consecutive patients underwent PD for periampullary tumor with routine bile culture and antibiotic prophylaxis with cefazolin. The postoperative course of 35 patients with infected bile (group B+) was compared with that of 44 patients with sterile bile (group B-). RESULTS: The distribution of tumors was comparable except for ampullary carcinoma, which was more frequent in group B+ patients (p = 0.001). Interventional biliary endoscopy was performed preoperatively in 80% of patients in group B+ versus 14% in group B- (p < 0.001), including 9 isolated sphincterotomies (20% versus 5%, p < 0.03) and 20 endoprosthesis insertions (57% versus 0%, p < 0.0001). More patients in group B+ were classified as American Society of Anesthesiologists (ASA) 2 (p = 0.04). Operative time and blood loss were similar in both groups. One patient died postoperatively (group B+). Overall morbidity was increased in group B+ (77% versus 59%, p = 0.05). Postoperative infectious complications, all demonstrated bacteriologically, were more frequent in group B+: (65% versus 37%, p = 0.003). In group B+, bile was polybacterial in 54% of patients and isolated microorganisms were resistant to cefazolin in 97%. In patients with infectious complications, the same germ was isolated in bile and another sample in 49%. CONCLUSIONS: In patients undergoing PD, bile infection is related to previous interventional biliary endoscopy in 80% of patients and is associated with an increased rate of postoperative infections. During PD for ampullary carcinoma or after interventional biliary endoscopy, a specific antibioprophylaxis should be evaluated. PMID- 16377503 TI - Longterm results and prognostic indicators after cryotherapy and hepatic arterial chemotherapy with or without resection for colorectal liver metastases in 224 patients: longterm survival can be achieved in patients with multiple bilateral liver metastases. AB - BACKGROUND: Only a minority of patients are suitable for liver resection for colorectal liver metastases. Cryotherapy was the first widely used ablative technique, achieving a median survival of more than 2 years. Patient selection is important, but the prognostic criteria have been controversial. The combined treatment modality of cryotherapy, resection, and hepatic arterial chemotherapy has been used in treating unresectable liver disease. Many centers would not surgically treat patients with multiple (five or more lesions) bilateral liver metastases. This series reports on longterm results after cryotherapy and hepatic arterial chemotherapy with or without liver resection in 224 patients with colorectal liver metastases, especially in patients with multiple bilateral liver disease, and identifies important prognostic determinants for survival. STUDY DESIGN: A longterm retrospective analysis was performed of prospectively collected clinical data of 224 patients with colorectal liver metastases who received cryotherapy and hepatic arterial chemotherapy with or without resection. Morbidity, mortality, recurrence, and survival rates were reported and Kaplan Meier and Cox regression analysis were used to identify prognostic indicators. RESULTS: Median length of followup was 26 months (range 1 to 130 months). Perioperative mortality rate was 0.4% and morbidity rate was 21%. Cryosite, remaining liver, and extrahepatic recurrence rates were 39%, 62%, and 67%, respectively. Median survival was 31 months (range 1 to 130 months) with 1-, 3-, and 5-year survival rates of 87%, 43%, and 23%, respectively. Ninety-one patients had five or more bilateral lesions. Median and 5-year survival was 31 months (2 to 88 months) and 26%, respectively. Four factors were independently associated with favorable survival outcomes: cryotherapy with resection, complete tumor eradication, low pre- and postoperative CEA levels. CONCLUSIONS: Cryotherapy and hepatic arterial chemotherapy with or without resection can achieve long survival advantage in patients with unresectable colorectal liver metastases. PMID- 16377504 TI - Predictors of failure after pancreaticoduodenectomy for ampullary carcinoma. AB - BACKGROUND: Complete resection offers the only potential cure for ampullary carcinoma. We analyzed factors that contribute to treatment failure and survival in patients who underwent pancreaticoduodenectomy for ampullary carcinoma. STUDY DESIGN: We retrospectively reviewed all patients who underwent pancreaticoduodenectomy between August 1994 and August 2003 for ampullary carcinoma. Demographic, clinical, and pathologic data were collected. Chi-square analysis was used for categorical data and the t-test was used for continuous variables. Kaplan-Meier analyses were compared using the log-rank test to examine patient survival. RESULTS: Forty-three patients (24 men) aged 63.7 +/- 11.4 years (standard deviation) were followed for a mean of 23.9 months (median 660 days, range 18 to 2,249 days). Jaundice (n = 33) and weight loss (n = 13) were the most common presenting symptoms. Stage (p < 0.01) and degree of differentiation (p < 0.029) were significant predictors of failure by univariate analysis. But only stage (p < 0.04) was a significant predictor by multivariate analysis. Further analysis revealed that nodal status (p < 0.001), but not tumor grade, was a significant predictor of treatment failure. Neither demographic nor clinical variables were significant predictors. Five-year overall and disease-free survival rates were 67.4% and 51.4%, respectively. Both metastases and disease recurrence had significant impact on patient survival. CONCLUSIONS: Tumor stage is associated with treatment failure after pancreaticoduodenectomy for ampullary carcinoma and may identify candidates for adjuvant therapy. Because an aggressive surgical approach can be adopted safely with the best chance for cure, we recommend that pancreaticoduodenectomy be offered to all patients with ampullary tumors when malignancy or dysplasia is in question. PMID- 16377505 TI - Survival, hemodynamics, and tissue oxygenation after head trauma. AB - BACKGROUND: The aims of this study were to describe the early time course of hemodynamic and tissue perfusion and oxygenation patterns in survivors and nonsurvivors after head injury; to suggest physiologic mechanisms responsible for the observed patterns; and to evaluate postinjury parameters that might be useful for treatment. The hypothesis was that reduced hemodynamics and tissue oxygenation and reduced arterial oxygen saturation affect outcomes. STUDY DESIGN: Sixty patients with head trauma were noninvasively monitored on arrival in the emergency department to assess the temporal hemodynamic patterns associated with head injury; patients who were brain dead were excluded because they have very different hemodynamic patterns. Cardiac index, mean arterial pressure, and heart rate were monitored to assess cardiac function, pulse oximetry to reflect changes in pulmonary function, and transcutaneous oxygen and carbon dioxide to reflect tissue perfusion function. Patients were stratified by inhospital survival outcomes, the Glasgow Coma Scale, and the presence or absence of associated somatic injuries. RESULTS: When all head injured patients were considered together, the predominant findings were high cardiac index, hypertension, mild tachycardia, normal pulmonary function, and reduced tissue oxygenation. The subset of survivors and those with high Glasgow Coma Scale had greater than normal cardiac index responses (4.02 +/- 0.01 (SEM) L/min/m2, p < 0.01 versus normal) and better tissue oxygenation (217 +/- 2 mmHg PtcO2/FiO2) than nonsurvivors (70 +/- 3 mmHg, p < 0.01) and those with low Glasgow Coma Scale (160 +/- 2, p < 0.05). Patterns of patients with associated somatic injuries were similar to those with isolated head injury. CONCLUSIONS: The study suggested that survivors' cardiac index, tissue oxygenation, and arterial oxygen saturation may be considered as markers of resuscitation. Nonsurvivors of head injury had normal blood flow with reduced tissue oxygenation that might have contributed to unfavorable outcomes. PMID- 16377506 TI - Transfusion increases the risk of postoperative infection after cardiovascular surgery. AB - BACKGROUND: Because of the immunomodulatory effects of transfusion, we attempted to identify factors associated with blood product use and determine the association of transfusion quantity with postoperative infection. STUDY DESIGN: We studied total perioperative transfusion of blood products for 15,592 cardiovascular operations performed from July 1998 to May 2003. Infection end points were septicemia/bacteremia (n=351, 2.2%) and superficial (n=353, 2.3%) and deep (n=212, 1.4%) sternal wound infections. Factors associated with blood product administration were used to form balancing scores to adjust for differences in patient characteristics among those receiving and not receiving blood products. RESULTS: Fifty-five percent of patients received packed red blood cells (RBC), 21% received platelets, 13% got fresh frozen plasma (FFP), and 3% got cryoprecipitate. Factors associated with RBC use included older age, female gender, higher New York Heart Association class, lower hematocrit, reoperation, and longer cardiopulmonary bypass time--all indicative of higher-risk patients. The more RBC units transfused, the higher was the occurrence of septicemia/bacteremia (p < 0.0001) and superficial (p=0.0007) and deep (p < 0.0001) sternal wound infection. Use of FFP (septicemia/bacteremia) and platelets (septicemia/bacteremia and deep sternal wound infection) mitigated against this association only slightly. CONCLUSIONS: Blood products tended to be used in the sickest patients. But after accounting for this, risk of infection increased incrementally with each unit of blood transfused. Although cause and effect cannot be established, results suggested that blood product transfusion is an independent risk factor for postoperative infection in cardiac surgical patients, blood products are more likely to be used in the sickest patients, and no amount of blood loss treated by transfusion is innocuous. PMID- 16377507 TI - Plasma D-dimer levels show correlation with number of lymph node metastases in patients with esophageal cancer. AB - BACKGROUND: Systemic activation of hemostasis and fibrolysis has been shown to be related to tumor progression in patients with malignancies such as lung cancer and colorectal cancer, but there has been no report of these clotting abnormalities in esophageal cancer. We investigated the clinical importance of measuring plasma levels of D-dimer (DD), which is a marker of the hypercoagulable stage, in preoperative patients with esophageal cancer. STUDY DESIGN: Preoperative plasma DD levels were measured in 96 patients with primary esophageal cancer who were scheduled for esophagectomy with lymphadenectomy without preoperative treatment at our hospital. Results were correlated with the clinicopathological findings. RESULTS: Significantly different plasma DD levels were found with respect to histologic T (p = 0.0015), histologic N (p < 0.0001), number of metastatic nodes (p < 0.0001), and histologic stages (p < 0.0001). The number of lymph node metastases (0/1 to 3/4 to 7/8-) was found to have the strongest association with DD level among the significant clinicopathologic factors (Spearman rank correlation 0.591, p < 0.0001). The most useful cut-off level of the plasma DD levels for diagnosis of lymph node metastasis was determined to be 0.4 microg/mL, with specificity and sensitivity for lymph node metastasis being 62.9% and 88.2%, respectively. CONCLUSIONS: Plasma DD levels are useful for assessing lymph node metastasis in patients with esophageal cancer and should be measured preoperatively. PMID- 16377508 TI - The surgeon and acts of civilian terrorism: radiation exposure and injury. PMID- 16377509 TI - Transarterial chemoembolization for hepatocellular carcinoma. PMID- 16377510 TI - Managing the spectrum of surgical pain: acute management of the chronic pain patient. PMID- 16377511 TI - Optimal nutrition, exercise, and hormonal therapy promote muscle anabolism in the elderly. PMID- 16377512 TI - Albert J and his surgeon: a historical reevaluation of the first parathyroidectomy. PMID- 16377513 TI - Military medicine at Little Bighorn. PMID- 16377514 TI - Venous air embolism from intravenous CT contrast administration. PMID- 16377515 TI - Dysphagia lusoria and aberrant right subclavian artery. PMID- 16377516 TI - Sinusoidal obstruction syndrome is a major feature of hepatic lesions associated with oxaliplatin neoadjuvant chemotherapy for liver colorectal metastases. PMID- 16377517 TI - Caudate hepatectomy for cancer. PMID- 16377518 TI - The aviation paradigm and surgical education. PMID- 16377520 TI - Schistosomal portal hypertension. PMID- 16377523 TI - Who should be vaccinated against human papillomavirus? PMID- 16377524 TI - Mycobacterium-associated immune reconstitution disease: macrophages running wild? PMID- 16377526 TI - Clinical examination versus Chlamydia trachomatis assays to guide antibiotic use in trachoma control programmes. PMID- 16377527 TI - Clinical examination and laboratory tests for estimation of trachoma prevalence in remote settings. PMID- 16377529 TI - Fear of flying. PMID- 16377531 TI - Genital human papillomavirus infection in men. AB - Genital human papillomavirus (HPV) infection, globally one of the most common sexually transmitted infections, is associated with cancers, genital warts, and other epithelial lesions. Although a consistent and coherent picture of the epidemiology and pathogenesis of genital HPV infections in women has developed over the past two decades, less is known about these infections in men. Available data suggest that, as with women, most genital HPV infections in men are symptomless and unapparent, and that HPV16 is probably the most frequently detected type. In populations of similar age, the prevalence of specific HPV types is usually lower in men than in women. Whether this observation relates to lower incidence or shorter duration of infection in men than in women has not yet been determined. Seroprevalence of specific anti-HPV antibodies also seems to be lower in men than in women of similar age, a difference that might be due to lower viral load, lower incidence or duration of infection or lower antibody responses, or both, in men compared with women. Differences in sexual behaviour may also be important predictors of genital HPV infection. With the anticipated availability of prophylactic HPV vaccines in the near future, it becomes increasingly important to understand the incidence and duration of HPV infections in men to develop cost-effective approaches to prevention through a combination of immunisation and promotion of risk-reduction strategies. PMID- 16377532 TI - Barriers to participation in HIV drug trials: a systematic review. AB - The enrolling of adequate participants into HIV experimental drug trials presents an important challenge. We systematically reviewed the literature to identify barriers and concerns amongst HIV patients to participation in HIV clinical drug trials. We reviewed studies for validity and content, and generated pooled estimates of the frequency with which patients identified particular issues by pooling across studies that presented results as proportions. We included three semi-structured interview studies, two open-ended questionnaires, and nine quantitative studies. Major barriers to participation included fear of side effects, distrust of researchers, general concerns about research design, interference in everyday life or changes in routine, and social discrimination. Results from the quantitative studies indicated that the most prevalent barriers were as follows: suspicions about the drug itself (53%, 95% CI 24-83%), patients were not informed or believed they were not eligible (38%, 25-50%), and travel or transport obstacles (39%, 21-57%). The findings of this study should aid drug trialists in developing strategies to maximise participation and cooperation in HIV clinical drug trials while adequately informing and protecting prospective participants. PMID- 16377533 TI - Mathematical modelling: a tool for hospital infection control. AB - Health-care-associated infections caused by antibiotic-resistant pathogens have become a menace in hospitals worldwide and infection control measures have lead to vastly different outcomes in different countries. During the past 6 years, a theoretical framework based on mathematical models has emerged that provides solid and testable hypotheses and opens the road to a quantitative assessment of the main obstructions that undermine current efforts to control the spread of health-care-associated infections in hospitals and communities. We aim to explain to a broader audience of professionals in health care, infection control, and health systems administration some of these models that can improve the understanding of the hidden dynamics of health-care-associated infections. We also appraise their usefulness and limitations as an innovative research and decision tool for control purposes. PMID- 16377534 TI - Beyond registration--measuring the public-health potential of new treatments for malaria in Africa. AB - Malaria claims over one million lives a year in some of the poorest countries of the world. Affected populations and governments cannot afford to pay for expensive new therapies. Most antimalarial treatments are purchased from local shops and administered in the home. These factors make for a complex set of requirements for any new treatment for malaria if a substantial reduction in mortality is ever to be achieved. Thankfully there are several treatments being developed, mostly within public-private partnerships. Typically, the goal of public-private partnerships is the granting of a product license, so work plans end after phase III trials. As these drugs will ultimately be used unsupervised, malaria control programme managers will require further data on safety and whether the drug is as efficacious when used outside of controlled clinical trials before allowing widespread use of these new products. These data need to be collected in highly specific phase IV programmes. We explain why public private partnerships should extend their development plans well beyond drug registration, and set out the requirements of such a programme. We aim to generate debate and discussion so that guidelines that are internationally accepted and adhered to can be developed not only for antimalarials but for all drugs that are being developed specifically for use in resource-poor settings. PMID- 16377535 TI - A new model to monitor the virological efficacy of antiretroviral treatment in resource-poor countries. AB - Monitoring the efficacy of antiretroviral treatment in developing countries is difficult because these countries have few laboratory facilities to test viral load and drug resistance. Those that exist are faced with a shortage of trained staff, unreliable electricity supply, and costly reagents. Not only that, but most HIV patients in resource-poor countries do not have access to such testing. We propose a new model for monitoring antiretroviral treatment in resource limited settings that uses patients' clinical and treatment history, adherence to treatment, and laboratory indices such as haemoglobin level and total lymphocyte count to identify virological treatment failure, and offers patients future treatment options. We believe that this model can make an accurate diagnosis of treatment failure in most patients. However, operational research is needed to assess whether this strategy works in practice. PMID- 16377536 TI - Gastric histoplasmosis. PMID- 16377538 TI - Use of intravenous immunoglobulin G (IVIG). AB - Intravenous immunoglobulin G (IVIG) has become increasingly important both as replacement therapy in primary and acquired humoral immunodeficiency and as an immunomodulatory therapy in autoimmune disease and transplantation. Multiple potential mechanisms for the effects of IVIG have now been recognized but the contribution of each mechanism in different diseases is uncertain. IVIG is generally well tolerated but serious side effects can occur and need to be addressed. IVIG has Food and Drug Administration (FDA) approval for a half dozen indications but these account for only about half the use of IVIG. This chapter reviews the development of IVIG for primary immunodeficiency, the evidence for efficacy of IVIG in autoimmune and inflammatory conditions, the risks associated with administration of IVIG, and steps that can be taken to minimize adverse events. PMID- 16377539 TI - Rh-immunoglobulin: Rh prophylaxis. PMID- 16377540 TI - Clinical uses of plasma and plasma fractions: plasma-derived products for hemophilias A and B, and for von Willebrand disease. AB - The use of plasma-derived factor products to treat hemophilia A, hemophilia B, and von Willebrand disease (vWD) has changed since the start of the human immunodeficiency virus (HIV) epidemic. The use of plasma-derived factor concentrates for hemophilias A and B has decreased in developed countries because of the availability of recombinant products. However, in developing countries, which encompass most of the world's hemophilia community, plasma-product-based therapy remains the backbone of treatment because of economic constraints. Viral attenuation strategies have resulted in a much safer product profile. vWD product selection is less complicated than for hemophilas A and B because plasma-derived products are the only choice for patients who are unresponsive or who cannot receive pharmacologic therapy. As the majority of patients in the world with hemophilias A, B and vWD are treated with plasma-derived clotting factors, the need for these safe and efficacious therapies will continue in the future. This chapter discusses safety strategies for plasma-derived clotting factor, its availability, economics, efficacy, and inhibitor formation. PMID- 16377541 TI - Management of factor VIII inhibitors. AB - The development of inhibitory alloantibodies to factor VIII is arguably one of the most severe and important complications of clotting factor concentrate exposure in haemophilia A. The development of an inhibitor compromises the ability to effectively manage haemorrhage, resulting in a greater rate of disability, complications and costs of therapy. This chapter briefly reviews the epidemiology, immunobiology, and laboratory evaluation of inhibitors. It discusses the therapeutic approach and management of inhibitors in various clinical settings and also focuses on inhibitor eradication practices (immune tolerance) and newer experimental strategies with potential clinical application for inhibitor prevention. PMID- 16377542 TI - Appraisal of the evidence for the clinical use of FFP and plasma fractions. AB - Randomised, controlled trials of good quality are a recognised means to robustly assess the efficacy of interventions in clinical practice. A systematic identification and appraisal of all randomised trials involving fresh frozen plasma (FFP) indicates that most clinical indications for FFP, as currently recommended by practice guidelines, are not supported by evidence from randomised trials. This chapter will largely consider the implications of some of the findings from this systematic review. Many published trials on the use of FFP have enrolled small numbers of patients, and provided inadequate information on the ability of the trial to detect meaningful differences in outcomes between the two patient groups. Other concerns about the design of the trials include the dose of FFP used, and the potential for bias; no studies had taken adequate account of the extent to which adverse effects might negate the clinical benefits of treatment with FFP. In addition, there is little evidence for the effectiveness of the prophylactic use of FFP. There is a pressing need to consider how best to develop new trials to determine the effectiveness of FFP. How this can be achieved can be illustrated by reference to studies of albumin in critical care. A recent, large and well-designed randomised trial (Saline versus Albumin Fluid Evaluation study; SAFE) in critical care found no evidence of an increase in mortality with the use of albumin compared to saline, which had been hypothesised in an earlier systematic review. How the study findings will actually now influence the clinical use of albumin remains to be seen. Although the SAFE trial showed no increase in mortality with albumin compared with saline, it is difficult to justify its use in critical care given its considerably greater cost. PMID- 16377543 TI - Clinical use of plasma and plasma fractions. AB - The use of plasma and plasma-derived products has always involved a careful balance of anticipated benefit versus risk. Risk reduction through pathogen inactivated products has been successful, but the expense of manufacture does not warrant widespread use. Although plasma has always had limited indications for use, these are often misunderstood or ignored in favor of received knowledge and tradition. Solid evidence from multiple trials support the limited indications for FFP described here and support products that target specific coagulation defects. PMID- 16377544 TI - Plasma and plasma products in the treatment of massive haemorrhage. AB - Massive haemorrhage requires the use of plasma products when it is accompanied by a coagulopathy or when the more than one blood volume has been lost and intractable bleeding continues. The coagulopathy results from haemorrhagic shock, hypothermia, and activation, consumption and dilution of coagulation factors. Plasma products have a critical role in maintaining sufficient levels of coagulation proteins to ensure haemostasis can occur. Fresh frozen plasma is a source of all coagulation proteins and is required when the prothrombin time and activated partial thromboplastin time exceed 1.5 times the normal control. Cryoprecipitate is the plasma product of choice if fibrinogen, the most critical coagulation protein, is required rapidly and to maintain levels at >1g/L. Prothrombin complex concentrates, monocomponent factor therapy and fibrin sealants each have a role in specific clinical settings. Recombinant factor VIIa has now been shown to have a role in massive haemorrhage. Randomised controlled trials are currently underway to determine the optimal dose and timing of its administration. The physiology and management of the coagulation disturbance using plasma products in the massive haemorrhage of specific clinical situations are described. PMID- 16377545 TI - Treating coagulopathy in liver disease with plasma transfusions or recombinant factor VIIa: an evidence-based review. AB - In severe liver disease, poor synthetic function leads to characteristic deficiencies in numerous coagulation factors, and plasma transfusions are frequently administered to treat or prevent bleeding. This chapter reviews the available English-language randomized controlled trials, evidence-based practice guidelines, and observational studies relevant to establishing criteria for plasma transfusions in liver disease. The alternatives of pathogen-inactivated plasmas and recombinant factor VIIa were also reviewed from this perspective. In current guidelines, plasma transfusions are justified when haemostasis is needed for bleeding or invasive procedures, and the prothrombin time (PT) or partial thromboplastin time (PTT) is >1.5 times normal (mid-normal or, for PTT, sometimes upper limit). Conversion of the PT to the International Normalized Ratio has not been validated in liver disease. Solvent-detergent or methylene-blue treatments alter various clotting factors, which might affect efficacy in liver disease. Recombinant factor VIIa improves laboratory clotting measurements, but reduction of bleeding is less well established to date. PMID- 16377546 TI - Plasma and plasma components in the management of disseminated intravascular coagulation. AB - A variety of clinical conditions can cause systemic activation of coagulation that ranges from insignificant laboratory changes to severe disseminated intravascular coagulation (DIC). DIC consists of a widespread systemic activation of coagulation, resulting in diffuse fibrin deposition in small and midsize vessels. There is compelling evidence from clinical and experimental studies that DIC is involved in the pathogenesis of microvascular dysfunction and contributes to organ failure. In addition, the massive and ongoing activation of coagulation can result in depletion of platelets and coagulation factors, which might cause bleeding. Recent insight into important pathogenetic mechanisms that might lead to DIC has resulted in novel preventive and therapeutic approaches to patients with sepsis and derangement of coagulation. Supportive strategies aimed at inhibition of coagulation activation might theoretically be justified and have been found beneficial in experimental and initial clinical studies. These strategies comprise inhibition of tissue factor-mediated activation of coagulation or restoration of physiological anticoagulant pathways. PMID- 16377547 TI - Pediatric hemostasis and use of plasma components. AB - Indications for fresh frozen plasma (FFP), once used routinely in the support of critically ill infants and children, have become more specific as evolving evidence has confirmed or disproved the efficacy of plasma in various circumstances. FFP is currently indicated to treat the coagulopathies of massive hemorrhage, liver failure and disseminated intravascular coagulation and sepsis. Whole blood reconstituted from FFP and packed red cells is the product of choice for exchange transfusion, as well as for circuit priming. In the US, FFP remains the only approved source of factors V, XI, protein C, protein S and plasminogen. Cryoprecipitate is used chiefly as a source of fibrinogen, factor VIII and factor XIII in consumptive coagulopathy; recombinant or viral inactivated plasma derivatives are preferred for congenital deficiencies of factor VIII and von Willebrand factor. Recombinant and highly purified, viral inactivated, plasma derived proteins are preferred over FFP for congenital and acquired deficiencies. This chapter reviews evidence to support the use of plasma and plasma derivatives for pediatric patients. PMID- 16377548 TI - Therapeutic apheresis: use of human serum albumin, fresh frozen plasma and cryosupernatant plasma in therapeutic plasma exchange. AB - In therapeutic plasma exchange, patient plasma is removed and a colloid replacement solution is infused in its stead. A solution of 4-5% human serum albumin in saline is the recommended replacement solution in most instances, even though it leads to transient mild deficiencies of most plasma proteins. Albumin solutions are pasteurized to inactivate viruses, carry a very low risk of febrile and allergic reactions, and are convenient to store and administer. Fresh frozen plasma, which must be type specific and needs to be ordered in advance and thawed before use, carries a higher risk of reactions; however, it replaces all plasma constituents and is appropriate for patients with thrombotic thrombocytopenic purpura or a pre-existing coagulopathy. Neither cryosupernatant plasma, which is relatively depleted of the proteins in cryoprecipitate, nor pooled plasma that has been virally inactivated with organic solvents and detergents has been shown to be superior to fresh frozen plasma for any indication. PMID- 16377549 TI - Risks and side effects of therapy with plasma and plasma fractions. AB - Transfusion of plasma can lead to adverse reactions or events. Immune-mediated reactions are most common--these include allergic and anaphylactic reactions, transfusion-related acute lung injury (TRALI) and haemolysis. They can range in severity from mild to fatal. Fluid overload and citrate toxicity can occur after rapid or massive transfusion. In developed countries, microbial transmission rates are low because of donor selection and testing. Pathogen reduction processes can be applied to either single-unit components (methylene blue) or plasma pools (solvent-detergent). They have the unwanted effect of reducing some coagulation factors but reduce viral transmission risk even further. Reactions associated with plasma products or fractions also include allergic reactions, although TRALI is rare. Viral transmission risk is very low because of the use of two independent viral inactivation steps. Different products have particular specific unwanted effects: intravenous immunoglobulin has been associated with thrombotic events, renal toxicity and aseptic meningitis; coagulation factors are associated with development of inhibitors and thrombotic events. The risk of transmission of variant Creutzfeldt-Jakob disease in both plasma components and pooled plasma products is as yet unknown. If anything, the low titre of prion infectivity in the blood of an infected individual (approximately 10 infectious units/ml) will be massively diluted by the thousands of units of plasma in the pool. Subsequent manufacturing processes also remove prions from the final product. PMID- 16377550 TI - Fibrin glues of human origin. AB - Wound healing is the response to injury and the process of tissue repair. Recent advances in cellular and molecular biology have greatly expanded our understanding of this process, which includes chemotaxis, production of matrix protein, cell replication, neovascularization and tissue remodelling. Tissue injury causes the disruption of blood vessels that is responsible for extravasation or haemorrhage of blood constituents and the first step of process in the platelet activation after exposure of collagen. Platelets initiate clotting through the coagulation system. When thrombin is formed, fibrinogen is transformed into fibrin; this is the real first step of wound healing. Fibrin glues reproduce this process and have been widely used in surgery to obtain haemostasis and expedite the process of wound healing. Immunogenicity of xenogenic products and risks of viral or prion disease transmission through commercial products have generated new interest in home-made autologous glues whose complexity was increased when platelets were added to coagulation proteins as a source of cytokines and growth factors. For the preparation of these products, the blood bankers took advantage of their usual technologies and several automated apparatuses are currently employed for the preparation of fibrinogen and thrombin concentrates. The machines for sequestration and multicomponent collection have been adapted for the concurrent procurement of plasma and platelets. A further level of complexity was introduced by the addition to glues of peripheral, bone marrow and cord-blood-derived stem cells to help or determine tissue regeneration. Instead of more new technologies, larger studies are now needed to quantify the benefits and clarify the optimal application for these products. PMID- 16377551 TI - Pathogen inactivation techniques. AB - The desire to rid the blood supply of pathogens of all types has led to the development of many technologies aimed at the same goal--eradication of the pathogen(s) without harming the blood cells or generating toxic chemical agents. This is a very ambitious goal, and one that has yet to be achieved. One approach is to shun the 'one size fits all' concept and to target pathogen-reduction agents at the Individual component types. This permits the development of technologies that might be compatible with, for example, plasma products but that would be cytocidal and thus incompatible with platelet concentrates or red blood cell units. The technologies to be discussed include solvent detergent and methylene blue treatments--designed to inactivate plasma components and derivatives; psoralens (S-59--amotosalen) designed to pathogen-reduce units of platelets; and two products aimed at red blood cells, S-303 (a Frale--frangible anchor-linker effector compound) and Inactine (a binary ethyleneimine). A final pathogen-reduction material that might actually allow one material to inactivate all three blood components--riboflavin (vitamin B2)--is also under development. The sites of action of the amotosalen (S-59), the S-303 Frale, Inactine, and riboflavin are all localized in the nucleic acid part of the pathogen. Solvent detergent materials act by dissolving the plasma envelope, thus compromising the integrity of the pathogen membrane and rendering it non-infectious. By disrupting the pathogen's ability to replicate or survive, its infectivity is removed. The degree to which bacteria and viruses are affected by a particular pathogen reducing technology relates to its Gram-positive or Gram-negative status, to the sporulation characteristics for bacteria, and the presence of lipid or protein envelopes for viruses. Concerns related to photoproducts and other breakdown products of these technologies remain, and the toxicology of pathogen-reduction treatments is a major ongoing area of investigation. Clearly, regulatory agencies have a major role to play in the evaluation of these new technologies. This chapter will cover the several types of pathogen-reduction systems, mechanisms of action, the inactivation efficacy for specific types of pathogens, toxicology of the various systems and the published research and clinical trial data supporting their potential usefulness. Due to the nature of the field, pathogen reduction is a work in progress and this review should be considered as a snapshot in time rather than a clear picture of what the future will bring. PMID- 16377552 TI - Plasma protein therapies: current and future perspectives. AB - Although early developments in immunology and haemostasis indicated the potential therapeutic application of plasma fractions, it was not until Cohn's development of a stable plasma protein solution for the treatment of battlefield injuries in the Second World War that the manufacture of plasma derivatives became part of industrial pharmaceutics. The resulting albumin product remained the mainstay of the plasma fractionation industry for the next 40 years but the sequential removal of 'unwanted' fractions en route to the final albumin product lent itself to the characterization and use of other products. By the 1970s, the harvesting of cryoprecipitate before the initiation of the Cohn fractionation scheme allowed access to products for treating the haemophilias, using simple precipitation and, from the 1970s, chromatographic methods to concentrate the coagulation factors. Further minor modifications allowed the administration of the immunoglobulin in high intravenous dosages, significantly extending the usage and indications of immunoglobulin products. By the 1980s, the needs for haemophilia A had made factor VIII, rather than albumin, the driver for plasma fractionation, and the advent of recombinant coagulation factors in the 1990s contributed to immunoglobulin assuming the position of the plasma procurement driver. In recent years, the plasma industry has developed a relatively large range of products but has also entered a period of consolidation as various pressures, such as technological innovations, infectious and other risks and quality requirements all of which are discussed in this review-decreased the demand of at least two of the three products that underpin the industry's current financial basis. At the same time, modern principles of evidence-based therapeutics are coming into play in the traditionally empirical base for most of the usage for plasma derivatives. The future use of plasma derivatives, at least in the developed world, will probably follow a different path to the one seen so far, and a sound understanding for the pathophysiology of the medical indications for plasma therapies should contribute to a continuing role for these medicines in modern therapeutics. PMID- 16377553 TI - Drumming up dollars for stem cell research. AB - With stringent U.S. federal restrictions on funding for human embryonic stem cell research still in place, states grapple with the issue of whether to fund such research themselves. PMID- 16377554 TI - Genetics in the age of systems biology. AB - Systems biology has become a fashionable label for a new generation of large scale experiments. This essay explores how classical approaches such as forward genetics fit into this emerging framework. PMID- 16377555 TI - 24p3 and its receptor: dawn of a new iron age? AB - 24p3 is a secreted protein that induces apoptosis in leukocytes. Recently, 24p3 has been shown to bind to iron-containing bacterial siderophores. In this issue of Cell, a receptor that internalizes 24p3 is identified. Internalization of iron bound to 24p3 prevents apoptosis. In contrast, internalization of the apo form of 24p3 that does not contain iron leads to cellular iron efflux and apoptosis via the proapoptotic protein Bim. PMID- 16377556 TI - Gene regulation by histone H1: new links to DNA methylation. AB - Linker histones of the H1 family are among the most abundant components of chromatin. In this issue of Cell, the effect of H1 downregulation on gene expression is examined. Although a 50% reduction of histone H1 levels in embryonic stem cells affects chromatin structure globally, the expression of very few genes is altered. Intriguingly, this study reveals a new link between H1 and DNA methylation. PMID- 16377557 TI - Stem cell research goes underground: the RETINOBLASTOMA-RELATED gene in root development. AB - Both cellular differentiation and stem cell maintenance must occur at the root apex in order to ensure the continuous growth of plant roots. In this issue of Cell, it is revealed that a canonical retinoblastoma pathway plays a crucial role in regulating the balance between differentiation and renewal of plant root stem cells. PMID- 16377558 TI - Pushing back the expansion of introns in animal genomes. AB - In a recent paper in Science, surveyed the position of introns in 30 genes of a marine annelid and showed that over 60% of the introns occupy positions identical to those in human homologs. In contrast, both human and marine annelid genes share only 30% of their introns with other invertebrates. These observations suggest that the common ancestor of most animal phyla had intron-rich genes and reinforce the notion that introns proliferated early in the evolutionary history of eukaryotes. PMID- 16377559 TI - Playing ping pong with pins: cortical and microtubule-induced polarity. AB - Cortical cell polarity controls mitotic spindle orientation in many cell types. In this issue of Cell, it is turned around and shown that the transfer of polarity information between the cortex and the spindle is not just one way. In Drosophila neuroblasts, the spindle also has polarizing activity on the cortex. PMID- 16377560 TI - Control of transcription through intragenic patterns of nucleosome composition. AB - Several recent papers show that differences in histone modification and the use of histone variants at the 5' and 3' ends of genes influence the location and kinetics of transcriptional initiation. The ultimate target of most epigenetic mechanisms may be the regulation of nucleosome occupancy, which in turn controls access to DNA at specific genomic locations. PMID- 16377561 TI - Fanconi anemia (cross)linked to DNA repair. AB - Fanconi anemia is characterized by hypersensitivity to DNA interstrand crosslinks (ICLs) and susceptibility to tumor formation. Despite the identification of numerous Fanconi anemia (FANC) genes, the mechanism by which proteins encoded by these genes protect a cell from DNA interstrand crosslinks remains unclear. The recent discovery of two DNA helicases that, when defective, cause Fanconi anemia tips the balance in favor of the direct involvement of the FANC proteins in DNA repair and the bypass of DNA lesions. PMID- 16377562 TI - Histone H1 depletion in mammals alters global chromatin structure but causes specific changes in gene regulation. AB - Linker histone H1 plays an important role in chromatin folding in vitro. To study the role of H1 in vivo, mouse embryonic stem cells null for three H1 genes were derived and were found to have 50% of the normal level of H1. H1 depletion caused dramatic chromatin structure changes, including decreased global nucleosome spacing, reduced local chromatin compaction, and decreases in certain core histone modifications. Surprisingly, however, microarray analysis revealed that expression of only a small number of genes is affected. Many of the affected genes are imprinted or are on the X chromosome and are therefore normally regulated by DNA methylation. Although global DNA methylation is not changed, methylation of specific CpGs within the regulatory regions of some of the H1 regulated genes is reduced. These results indicate that linker histones can participate in epigenetic regulation of gene expression by contributing to the maintenance or establishment of specific DNA methylation patterns. PMID- 16377563 TI - MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. AB - Histone variant H2AX phosphorylation in response to DNA damage is the major signal for recruitment of DNA-damage-response proteins to regions of damaged chromatin. Loss of H2AX causes radiosensitivity, genome instability, and DNA double-strand-break repair defects, yet the mechanisms underlying these phenotypes remain obscure. Here, we demonstrate that mammalian MDC1/NFBD1 directly binds to phospho-H2AX (gammaH2AX) by specifically interacting with the phosphoepitope at the gammaH2AX carboxyl terminus. Moreover, through a combination of biochemical, cell-biological, and X-ray crystallographic approaches, we reveal the molecular details of the MDC1/NFBD1-gammaH2AX complex. These data provide compelling evidence that the MDC1/NFBD1 BRCT repeat domain is the major mediator of gammaH2AX recognition following DNA damage. We further show that MDC1/NFBD1-gammaH2AX complex formation regulates H2AX phosphorylation and is required for normal radioresistance and efficient accumulation of DNA-damage response proteins on damaged chromatin. Thus, binding of MDC1/NFBD1 to gammaH2AX plays a central role in the mammalian response to DNA damage. PMID- 16377564 TI - Termination factor-mediated DNA loop between termination and initiation sites drives mitochondrial rRNA synthesis. AB - The human mitochondrial transcription termination factor mTERF plays a central role in the control of heavy-strand rDNA transcription by promoting initiation, besides termination, of this transcription. However, until now, the mechanism underlying this stimulation of transcription by mTERF was not understood. In the present work, addition of mTERF to a HeLa cell mitochondrial lysate-based reaction mixture containing an artificial rDNA template did indeed specifically stimulate rDNA transcription. This stimulation required that mTERF be simultaneously bound to the rDNA transcription termination and initiation sites in the same molecule, thus forming a loop. Most significantly, a double binding of mTERF to the rDNA molecule, with resulting loop formation, was also shown in vivo. These results strongly suggest that, to satisfy the need for high rate of rDNA transcription, human mitochondrial rRNA synthesis involves mTERF-mediated rDNA looping that promotes recycling of the transcription machinery. PMID- 16377565 TI - In vitro analysis of huntingtin-mediated transcriptional repression reveals multiple transcription factor targets. AB - Transcriptional dysregulation has emerged as a potentially important pathogenic mechanism in Huntington's disease, a neurodegenerative disorder associated with polyglutamine expansion in the huntingtin (htt) protein. Here, we report the development of a biochemically defined in vitro transcription assay that is responsive to mutant htt. We demonstrate that both gene-specific activator protein Sp1 and selective components of the core transcription apparatus, including TFIID and TFIIF, are direct targets inhibited by mutant htt in a polyglutamine-dependent manner. The RAP30 subunit of TFIIF specifically interacts with mutant htt both in vitro and in vivo to interfere with formation of the RAP30-RAP74 native complex. Importantly, overexpression of RAP30 in cultured primary striatal cells protects neurons from mutant htt-induced cellular toxicity and alleviates the transcriptional inhibition of the dopamine D2 receptor gene by mutant htt. Our results suggest a mutant htt-directed repression mechanism involving multiple specific components of the basal transcription apparatus. PMID- 16377566 TI - Crystal structures of the ribosome in complex with release factors RF1 and RF2 bound to a cognate stop codon. AB - During protein synthesis, translational release factors catalyze the release of the polypeptide chain when a stop codon on the mRNA reaches the A site of the ribosome. The detailed mechanism of this process is currently unknown. We present here the crystal structures of the ribosome from Thermus thermophilus with RF1 and RF2 bound to their cognate stop codons, at resolutions of 5.9 Angstrom and 6.7 Angstrom, respectively. The structures reveal details of interactions of the factors with the ribosome and mRNA, including elements previously implicated in decoding and peptide release. They also shed light on conformational changes both in the factors and in the ribosome during termination. Differences seen in the interaction of RF1 and RF2 with the L11 region of the ribosome allow us to rationalize previous biochemical data. Finally, this work demonstrates the feasibility of crystallizing ribosomes with bound factors at a defined state along the translational pathway. PMID- 16377567 TI - A microRNA mediates EGF receptor signaling and promotes photoreceptor differentiation in the Drosophila eye. AB - A critical question about signal transduction is how weak or transient activation of signaling pathways achieves a robust and long-term switch in gene expression. We report that a microRNA is part of a mechanism that makes cells sensitive to signals in the Drosophila eye. Expression of miR-7 is activated in cells as they begin differentiating into photoreceptors. This is dependent on EGF receptor (EGFR) signaling that triggers ERK-mediated degradation of the transcription factor Yan. In nonstimulated cells, Yan represses miR-7 transcription, whereas miR-7 RNA represses Yan protein expression in photoreceptors, by binding to sequences within its mRNA 3'UTR. We propose that reciprocal negative feedback between Yan and miR-7 ensures mutually exclusive expression, with Yan in progenitor cells and miR-7 in photoreceptor cells. Expression is switched when EGFR signaling transiently triggers Yan degradation. This two-tiered mechanism explains how signal transduction activity can robustly generate a stable change in gene-expression patterns. PMID- 16377568 TI - Endogenous siRNAs derived from a pair of natural cis-antisense transcripts regulate salt tolerance in Arabidopsis. AB - In higher eukaryotes, miRNAs and siRNAs guide translational inhibition, mRNA cleavage, or chromatin regulation. We found that the antisense overlapping gene pair of Delta(1)-pyrroline-5-carboxylate dehydrogenase (P5CDH), a stress-related gene, and SRO5, a gene of unknown function, generates two types of siRNAs. When both transcripts are present, a 24-nt siRNA is formed by a biogenesis pathway dependent on DCL2, RDR6, SGS3, and NRPD1A. Initial cleavage of the P5CDH transcript guided by the 24-nt siRNA establishes a phase for the subsequent generation of 21-nt siRNAs by DCL1 and further cleavage of P5CDH transcripts. The expression of SRO5 is induced by salt, and this induction is required to initiate siRNA formation. Our data suggest that the P5CDH and SRO5 proteins are also functionally related, and that the P5CDH-SRO5 gene pair defines a mode of siRNA function and biogenesis that may be applied to other natural cis-antisense gene pairs in eukaryotic genomes. PMID- 16377569 TI - A cell-surface receptor for lipocalin 24p3 selectively mediates apoptosis and iron uptake. AB - The lipocalin mouse 24p3 has been implicated in diverse physiological processes, including apoptosis due to interleukin-3 (IL-3) deprivation and iron transport. Here we report cloning of the 24p3 cell-surface receptor (24p3R). Ectopic 24p3R expression confers on cells the ability to undergo either iron uptake or apoptosis, dependent upon the iron content of the ligand: Iron-loaded 24p3 increases intracellular iron concentration without promoting apoptosis; iron lacking 24p3 decreases intracellular iron levels, which induces expression of the proapoptotic protein Bim, resulting in apoptosis. Intracellular iron delivery blocks Bim induction and suppresses apoptosis due to 24p3 addition or IL-3 deprivation. We find, unexpectedly, that the BCR-ABL oncoprotein activates expression of 24p3 and represses 24p3R expression, rendering BCR-ABL(+) cells refractory to secreted 24p3. By inhibiting BCR-ABL, imatinib induces 24p3R expression and, consequently, apoptosis. Our results reveal an unanticipated role for intracellular iron regulation in an apoptotic pathway relevant to BCR-ABL induced myeloproliferative disease and its treatment. PMID- 16377570 TI - Dietary and genetic control of glucose transporter 2 glycosylation promotes insulin secretion in suppressing diabetes. AB - Pancreatic beta cell-surface expression of glucose transporter 2 (Glut-2) is essential for glucose-stimulated insulin secretion, thereby controlling blood glucose homeostasis in response to dietary intake. We show that the murine GlcNAcT-IVa glycosyltransferase is required for Glut-2 residency on the beta cell surface by constructing a cell-type- and glycoprotein-specific N-glycan ligand for pancreatic lectin receptors. Loss of GlcNAcT-IVa, or the addition of glycan ligand mimetics, attenuates Glut-2 cell-surface half-life, provoking endocytosis with redistribution into endosomes and lysosomes. The ensuing impairment of glucose-stimulated insulin secretion leads to metabolic dysfunction diagnostic of type 2 diabetes. Remarkably, the induction of diabetes by chronic ingestion of a high-fat diet is associated with reduced GlcNAcT-IV expression and attenuated Glut-2 glycosylation coincident with Glut-2 endocytosis. We infer that beta cell glucose-transporter glycosylation mediates a link between diet and insulin production that typically suppresses the pathogenesis of type 2 diabetes. PMID- 16377571 TI - Microtubule-induced Pins/Galphai cortical polarity in Drosophila neuroblasts. AB - Cortical polarity regulates cell division, migration, and differentiation. Microtubules induce cortical polarity in yeast, but few examples are known in metazoans. We show that astral microtubules, kinesin Khc-73, and Discs large (Dlg) induce cortical polarization of Pins/Galphai in Drosophila neuroblasts; this cortical domain is functional for generating spindle asymmetry, daughter cell-size asymmetry, and distinct sibling fates. Khc-73 localizes to astral microtubule plus ends, and Dlg/Khc-73 and Dlg/Pins coimmunoprecipitate, suggesting that microtubules induce Pins/Galphai cortical polarity through Dlg/Khc-73 interactions. The microtubule/Khc-73/Dlg pathway acts in parallel to the well-characterized Inscuteable/Par pathway, but each provides unique spatial and temporal information: The Inscuteable/Par pathway initiates at prophase to coordinate neuroblast cortical polarity with CNS tissue polarity, whereas the microtubule/Khc-73/Dlg pathway functions at metaphase to coordinate neuroblast cortical polarity with the mitotic spindle axis. These results identify a role for microtubules in polarizing the neuroblast cortex, a fundamental step for generating cell diversity through asymmetric cell division. PMID- 16377572 TI - The RETINOBLASTOMA-RELATED gene regulates stem cell maintenance in Arabidopsis roots. AB - The maintenance of stem cells in defined locations is crucial for all multicellular organisms. Although intrinsic factors and signals for stem cell fate have been identified in several species, it has remained unclear how these connect to the ability to reenter the cell cycle that is one of the defining properties of stem cells. We show that local reduction of expression of the RETINOBLASTOMA-RELATED (RBR) gene in Arabidopsis roots increases the amount of stem cells without affecting cell cycle duration in mitotically active cells. Conversely, induced RBR overexpression dissipates stem cells prior to arresting other mitotic cells. Overexpression of D cyclins, KIP-related proteins, and E2F factors also affects root stem cell pool size, and genetic interactions suggest that these factors function in a canonical RBR pathway to regulate somatic stem cells. Expression analysis and genetic interactions position RBR-mediated regulation of the stem cell state downstream of the patterning gene SCARECROW. PMID- 16377573 TI - Stem cells are units of natural selection in a colonial ascidian. AB - Stem cells are highly conserved biological units of development and regeneration. Here we formally demonstrate that stem cell lineages are also legitimate units of natural selection. In a colonial ascidian, Botryllus schlosseri, vascular fusion between genetically distinct individuals results in cellular parasitism of somatic tissues, gametes, or both. We show that genetic hierarchies of somatic and gametic parasitism following fusion can be replicated by transplanting cells between colonies. We prospectively isolate a population of multipotent, self renewing stem cells that retain their competitive phenotype upon transplantation. Their single-cell contribution to either somatic or germline fates, but not to both, is consistent with separate lineages of somatic and germline stem cells or pluripotent stem cells that differentiate according to the niche in which they land. Since fusion is restricted to individuals that share a fusion/histocompatibility allele, these data suggest that histocompatibility genes in Botryllus evolved to protect the body from parasitic stem cells usurping asexual or sexual inheritance. PMID- 16377574 TI - Apoptosis, cell proliferation and serotonin immunoreactivity in gut of Liza aurata from natural heavy metal polluted environments: preliminary observations. AB - In the present paper, the effect of natural environment non-lethal heavy metal concentration on cell renewal of Liza aurata intestinal epithelium, was studied by the TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling) method and anti-PCNA (proliferating cell nuclear antigen) immunohistochemistry, in order to detect, respectively, apoptosis and cell proliferation. In addition, the presence and distribution of the cell renewal regulator, serotonin, was immunohistochemically investigated. In order to reduce variability, only immature specimens were considered. The results indicated that in the control specimens from non-polluted areas, the PCNA immunoreactive nuclei of the proximal intestinal epithelium were only located at the bottom of the intestinal folds, together with a few TUNEL-positive nuclei, and goblet mucous differentiated cells. In the specimens from polluted areas, the number of PCNA immunoreactive cells was greatly enhanced, and they extended along the mid portion of the intestinal folds; the number of TUNEL-positive nuclei was enhanced as well, but they were almost exclusively detected in the third apical portion of the intestinal folds. Serotonin immunoreactive nerve elements were more frequently detected in the intestinal wall of L. aurata specimens from polluted areas, and besides that, some serotonin immunoreactive endocrine cells were also present. Variations in distribution and frequency of TUNEL-positive nuclei, PCNA immunoreactive nuclei, and serotonin immunoreactivity put in evidence an alteration of cell renewal with an enhancement of cell proliferation, probably leading to morphological intestinal fold changes. PMID- 16377575 TI - Presence and distribution of serotonin immunoreactivity in the cyprids of the barnacle Balanus amphitrite. AB - In this work, the presence and distribution of serotonin in the cyprid of the barnacle Balanus amphitrite were investigated by immunohistochemical methods. Serotonin-like immuno-reactive neuronal cell bodies were detected in the central nervous system only. Various clusters of immunoreactive neuronal cell bodies are distributed in the brain (protocerebrum, deutocerebrum, optical lobes), and at least, four pairs of neuronal cell bodies were detected in the centrally positioned neuropil of the posterior ganglion. Rich plexuses of immunoreactive nerve fibers in the neuropil area were also observed. Furthermore, bundles of strongly immunoreactive nerve fibers surrounding the gut wall were localized, and immunoreactive nerve terminals in the antennules and compound eyes were observed. These data demonstrate the presence of a serotonin-like immunoreactive substance in the barnacle cyprids; furthermore, its immunolocalization in the cephalic nerve terminals allows us to postulate the involvement of this bioactive molecule in substrate recognition during the settlement process. PMID- 16377576 TI - A new monoclonal antibody against DNA ligase I is a suitable marker of cell proliferation in cultured cell and tissue section samples. AB - The extensive characterization of the replicative human DNA ligase I (LigI) undertaken in the last decade demonstrated that the level of this protein strongly correlates with the rate of cell proliferation. This may allow to expand the repertoire of clinical biomarkers for the analysis of cell proliferation. We have produced a new monoclonal antibody (5H5) against LigI and exploited it as cell proliferation marker in Western blotting and immunofluorescence as well as in immunohistochemistry on paraffin tissue sections. The Western blot analysis showed that the LigI level detected by 5H5 antibody is high in all proliferating cells. On the contrary the protein is down regulated in resting human fibroblast and peripheral blood lymphocytes. Immunofluorescence analysis on cultured HeLa cells showed that 5H5 antibody labels all proliferating cells and displays the same staining pattern of BrdU in S-phase nuclei. Finally the analysis of serial sections of inflamed tonsils and NHL lymph nodes (either frozen or paraffin embedded) demonstrated that 5H5 marks the same population of cells as the Ki-67 antibody. Our results demonstrate that 5H5 antibody is a valuable tool for labeling proliferating cells that can be conveniently used in Western blotting, immunocytochemistry and immunohistochemistry. PMID- 16377577 TI - Emerin increase in regenerating muscle fibers. AB - The fate of emerin during skeletal muscle regeneration was investigated in an animal model by means of crush injury. Immunofluorescence, immunoblotting and mRNA analysis demonstrated that emerin level is increased in regenerating rat muscle fibers with respect to normal mature myofibers. This finding suggests an involvement of emerin during the muscle fiber regeneration process, in analogy with its reported involvement in muscle cell differentiation in vitro. The impairment of skeletal muscle physiological regeneration or reorganization could be a possible pathogenetic mechanism for Emery Dreifuss muscular dystrophy. PMID- 16377578 TI - Basal lamina structural alterations in human asymmetric aneurismatic aorta. AB - Basal lamina (BL) is a crucial mechanical and functional component of blood vessels, constituting a sensor of extracellular microenvironment for endothelial cells and pericytes. Recently, an abnormality in the process of matrix microfibrillar component remodeling has been advocated as a mechanism involved in the development of aortic dilation. We focused our attention on BL composition and organization and studied some of the main components of the Extracellular Matrix such as Tenascin, Laminins, Fibronectin, type I, III and IV Collagens. We used surgical fragments from 27 patients, submitted to operation because of aortic root aneurysm and 5 normal aortic wall specimens from heart donors without any evidence for aneurysmal or atherosclerotic diseases of the aorta. Two samples of aortic wall were harvested from each patient, proximal to the sinotubular junction at the aortic convexity and concavity. Each specimen was processed both for immunohistochemical examination and molecular biology study. We compared the convexity of each aortic sample with the concavity of the same vessel, and both of them with the control samples. The synthesis of mRNA and the levels of each protein were assessed, respectively, by RT-PCR and Western Blot analysis. Immunohistochemistry elucidated the organization of BL, whose composition was revealed by molecular biology. All pathological samples showed a wall thinner than normal ones. Basal lamina of the aortic wall evidentiated important changes in the tridimensional arrangement of its major components which lost their regular arrangement in pathological specimens. Collagen I, Laminin alpha2 chain and Fibronectin amounts decreased in pathological samples, while type IV Collagen and Tenascin synthesis increased. Consistently with the common macroscopic observation that ascending aorta dilations tend to expand asymmetrically, with prevalent involvement of the vessel convexity and relative sparing of the concavity, Collagen type IV is more evident in the concavity and Tenascin in the convexity. PMID- 16377579 TI - Expression of ATP7B in normal human liver. AB - ATP7B is a copper transporting P-type ATPase, also known as Wilson disease protein, which plays a key role in copper distribution inside cells. Recent experimental data in cell culture have shown that ATP7B putatively serves a dual function in hepatocytes: when localized to the Golgi apparatus, it has a biosynthetic role, delivering copper atoms to apoceruloplasmin; when the hepatocytes are under copper stress, ATP7B translocates to the biliary pole to transport excess copper out of the cell and into the bile canaliculus for subsequent excretion from the body via the bile. The above data on ATP7B localization have been mainly obtained in tumor cell systems in vitro. The aim of the present work was to assess the presence and localization of the Wilson disease protein in the human liver. We tested immunoreactivity for ATP7B in 10 human liver biopsies, in which no significant pathological lesion was found using a polyclonal antiserum specific for ATP7B. In the normal liver, immunoreactivity for ATP7B was observed in hepatocytes and in biliary cells. In the hepatocytes, immunoreactivity for ATP7B was observed close to the plasma membrane, both at the sinusoidal and at the biliary pole. In the biliary cells, ATP7B was localized close to the cell membrane, mainly concentrated at the basal pole of the cells. The data suggest that, in human liver, ATP7B is localized to the plasma membrane of both hepatocytes and biliary epithelial cells. PMID- 16377580 TI - Atrial natriuretic peptide secretion during development of the rat supraoptic nucleus. AB - Since a relationship between atrial natriuretic peptide and oxytocin was recently demonstrated in the heart (Gutkowska et al., 1997), the aim of this study was to determine whether a relationship between the two peptides is present also in the rat hypothalamus. For this purpose, we measured ANP-ontogeny in the rat hypothalamus immunohistochemically and compared it with oxytocin-ontogeny which we previously studied. The results showed that the ANP-peptide and mRNA-ANP start at the 18th day of the fetal life. Our earlier data for oxytocin in the rat hypothalamus showed that only mRNA-oxytocin appeared the 18th day of foetal life (Farina Lipari et al., 2001); thus, at the 18th day of foetal life, mRNA-ANP, ANP peptide and mRNA-oxytocin are present. We conclude that in the hypothalamus, differently from that in the heart, ANP might play a role on the synthesis of the oxytocin since ANP and its mRNA appear earlier than oxytocin. PMID- 16377581 TI - Occurrence of nitric oxide synthase in Megoura viciae Buckton (Homoptera, Aphididae): an histochemical and immunohistochemical localisation. AB - Nitric oxide (NO) is known to be involved in many physiological reactions of insects. We analysed NOS localisation in aphids of the species Megoura viciae by means of histochemical reaction for the NADPH-diaphorase activity and immunohistochemical methods for uNOS, nNOS and iNOS. The obtained data provided a complex and peculiar pattern of NOS distribution in cells and tissue of M. viciae. The histochemical reaction for NADPH-diaphorase was an indicative, but not exact marker of NOS localisation in aphids. The use of anti uNOS antiserum (frequently applied in insects) was of limited value in our specimens, whereas more satisfactory results were obtained with anti nNOS and iNOS antisera of human origin. The results of Western blot analysis confirmed the immunohistochemical ones, showing an aphid protein that reacted strongly with the polyclonal antibody anti-iNOS and anti-nNOS while a similar protein band was weakly immunoreactive with the polyclonal antibody anti-uNOS. Our results suggest that NO, prevalently synthesised by calcium/calmodulin-dependent isoform, plays important physiological roles both in adult and embryological stages of aphids. The data of principal interest was NOS presence in bacteriocytes, cells that host symbiotic prokaryotes belonging to the species Buchnera aphidicola, and in nuclei of adipocytes and gut cells. PMID- 16377582 TI - Liposomal pegylated doxorubicin versus low-dose recombinant interferon Alfa-2a in the treatment of advanced classic Kaposi's sarcoma; retrospective analysis of three German centers. AB - BACKGROUND: Classic Kaposi's sarcoma (KS) is a rare neoplasm, predominantly occurring in older subjects of Eastern Europe or Mediterranean descent. While single lesions may be treated by simple excision, laser therapy, cryotherapy, or intralesional therapy, advanced or disseminated disease requires systemic treatment. Several studies reported the effectiveness of pegylated liposomal doxorubicin (PLD) and low-dose recombinant interferon alfa-2a (IFNalpha) in the treatment of AIDS-associated KS. OBJECTIVE: The aim of this retrospective analysis of three German centers was to compare the effectiveness and tolerability of PLD with IFNalpha in patients with advanced classic KS. METHODS: Retrospective analysis of 18 Caucasian patients who had been treated for histologically proven classic KS, with either with PLD or IFNalpha was performed. Twelve patients received 20 mg/m2 of PLD monthly, and the number of cycles was adapted to the clinical response. Dose reduction or increased cycle length was conducted if toxicity intervened. In 6 patients, 3 million U of IFNalpha was injected subcutaneously 3 times a week. IFNalpha -therapy was adapted according to the clinical response. RESULTS: In the 12 KS patients treated with PLD, complete response (CR) was achieved in 8 (67 percent), major response (MR) in 3 (25 percent), and minor response (mR) in 1 (8 percent). Stable disease (SD) or progression of disease (PD) was not observed. An initial response was noted after 4-16 weeks of treatment (mean 8.6 weeks), the mean cumulative dose of PLD was 571.5 mg/m2 (range, 40 to 1496 mg/m2), and the mean follow-up was 13 months. Neutropenia (33 percent) related to PLD was the most common adverse event (4/12). Vomiting occurred in 3 (25 percent) patients; none of these were severe. Six patients were treated with IFNalpha. MR was achieved in 1 (17 percent), mR in 4 (67 percent) and SD in 1 of 6 patients (17 percent), neither had CR or PD. An initial response was observed after 8-17 weeks of treatment (mean 12.7 weeks). Fever occurred in 4 patients (67 percent). Flu-like symptoms in 3 patients (50 percent) related to IFNalpha were the most common adverse events. Mean follow-up was 6.3 months. The differences in response to treatment between PLD and IFNalpha, in general, were significant with p < 0.05 (T-test for independent samples). Comparing weeks to respond and treatment efficiency data were significant with p < 0.001 (Fisher's exact): response to PLD was up to one-third faster than IFNalpha. Calculating different stages of response (MR, CR, etc.), PLD also was clearly superior (p = 0.018) to IFNalpha (Fisher's exact). CONCLUSION: This retrospective analysis of patients with classic KS confirms the efficacy and safety of PLD. The benefits of PLD, including the monthly application, the high response even after previous treatments have failed, and the low rate of side effects even in elderly individuals, outweigh the risks. PLD is superior to IFNalpha and should be considered as an promising option in the treatment of advanced classic KS. PMID- 16377583 TI - STK15 gene overexpression, centrosomal amplification, and chromosomal instability in the absence of STK15 mutations in laryngeal carcinoma. AB - Centrosomes regulate cell division by forming bipolar mitotic spindles and, thus, play an essential role in the maintenance of chromosomal stability. Centrosomal amplification has been found commonly among tumor cells. Previous studies have suggested that a STK15 (serine/threonine kinase 15) gene can induce centrosomal amplification, chromosomal instability, and cell transformation. To investigate the role of STK15 gene abnormalities in the occurrence of centrosomal amplification and chromosomal instability, a combinatory approach has been taken to investigate the expression level and point mutations of the STK15 and centrosomal/chromosomal aberrations among 72 cases of laryngeal squamous cell carcinoma and a representative Hep-2 cell line. Although no mutation was detected within its exons 6 or 7, overexpression of STK15 has been found in 47 cases (65 percent) as well as in the Hep-2 cell line; for the latter apparent centrosomal amplification also has been noted, with the number of centrosomes within a single cell varying between 1 and 7 and the proportion of cells with amplified centrosomes reaching 11 approximately 23 percent. Karyotype analysis of Hep-2 cell line has suggested common occurrence of chromosomal aberrations, with the number of chromosomes ranging between 43 and 84, modal number between 69 and 74, and structural aberrations, represented by 13 marker chromosomes, including translocations, deletions, and isochromosomes found in various subclones. Our results suggest that in Hep-2 cell line overexpression of STK15 gene may cause centrosomal amplification thereby result in chromosomal instability through abnormal mitosis. Detection of STK15 overexpression in laryngeal carcinoma has led us to propose that the above may be one of the mechanisms underlying laryngeal carcinogenesis. PMID- 16377584 TI - Phase I clinical trial of topotecan and pegylated liposomal doxorubicin. AB - BACKGROUND: The objective of this study was to determine the feasibility and maximum tolerated dose (MTD) of combination topotecan and pegylated liposomal doxorubicin (PLD) administered in 4- or 3-week cycles in patients with advanced or refractory solid tumors. PATIENTS AND METHODS: Patients were treated with intravenous topotecan (0.75-1.25 mg/m2) for 3 days followed by PLD (25-40 mg/m2) on Day 4. The following dose combinations (topotecan/PLD, mg/m2) were explored: 0.75/40, 1.0/40, and 1.25/40 every 28 days; and 1.0/25 and 1.0/30 every 21 days. RESULTS: Thirty-two patients were enrolled, and all had received prior chemotherapy. Most (84 percent) patients had ovarian cancer. A total of 157 cycles (median, 4 cycles; range, 1-19 cycles) of chemotherapy were administered. Dose-limiting toxicities were Grade 4 neutropenia and death at dose level 3 (1.25/40 mg/m2 every 28 days), and neutropenic fever, Grade 3 stomatitis, and Grade 3 peripheral neuropathy (all in one patient) at dose level 5 (1/30 mg/m2 every 21 days). Myelosuppression was the most common serious toxicity. Twenty-six patients were evaluable for response and 7 (27 percent) had partial responses. All responses were seen in patients with ovarian cancer. CONCLUSIONS: This combination is feasible and well tolerated; encouraging activity was observed in heavily pretreated patients with ovarian cancer. The recommended regimens for a Phase II study are topotecan 1.0 mg/m2 on Days 1-3 followed by PLD 40 mg/m2 on Day 4 of a 28-day cycle, and topotecan 1.0 mg/m2 on Days 1-3 and PLD 30 mg/m2 on Day 4 of a 21-day cycle. PMID- 16377585 TI - Phase I study of CT-2103, a polymer-conjugated paclitaxel, and carboplatin in patients with advanced solid tumors. AB - PURPOSE: The primary objective of this study was to determine the maximum tolerated dose (MTD) of CT-2103 (poly L-glutamic acid-paclitaxel) in combination with carboplatin in patients with histologically proven solid tumors that were either refractory to conventional treatment or for which no conventional therapy was available. PATIENTS AND METHODS: Twenty-two adult patients with advanced solid tumors were treated in this dose escalation study. Patients were treated every 21 days with CT-2103 at 175, 210, 225, or 250 mg/m2 (doses expressed as units of conjugated-paclitaxel) via 10-20 minute intravenous (IV) infusion, followed one hour later with carboplatin administered at AUC 5 or 6 via 30 minute IV infusion. No prophylaxis for hypersensitivity was administered with initial treatment. Doses were administered every 21 days until progressive disease or dose-limiting toxicity (DLT) was observed. Toxicity was evaluated using NCI Common Toxicity Criteria for Adverse Events v2.0 (CTCAE v2.0); response to treatment was evaluated using Response Criteria in Solid Tumors (RECIST). RESULTS: The MTD was determined to be 225 mg/m2. DLTs observed at 250 mg/m2 were neutropenia and thrombocytopenia. No hypersensitivity reactions were observed. Three patients achieved partial responses (PR). Fifteen patients received at least 3 cycles of treatment without observation of progressive disease. Median survival time was 5.9 months. Patients that demonstrated partial responses were all ovarian cancer patients that had previously failed paclitaxel therapy. The only Grade 4, nonhematologic treatment-related toxicity was febrile neutropenia. Grade 4 neutropenia (9 patients) was observed across all dose groups. Twelve patients developed thrombocytopenia (Grade 3/4) while receiving combination therapy. All had resolution of thrombocytopenia with discontinuation of carboplatin, suggesting that carboplatin, and not CT-2103, contributed mainly to platelet toxicity. CONCLUSION: CT-2103 administered at 225 mg/m2 every 21 days in combination with carboplatin administered at AUC 6 has a manageable safety profile in patients with solid tumors; further clinical investigation is recommended, especially in patients with ovarian or non-small cell lung cancer. PMID- 16377586 TI - Hepatic arteriogram for gestational trophoblastic tumor: is it useful? AB - The presence of liver metastasis will be staged as IV in the FIGO 1992 Gestational Trophoblastic Tumor (GTT) staging. This study was to determine the role of hepatic arteriogram (HAG) in the management of GTT. It is a retrospective analysis of 309 patients treated from 1981 to 2001. Patients were restaged according to the FIGO 1992 classification with or without taking into account the HAG result. Outcome measures including mortality, drug resistance and recurrence of disease, as well as treatment with and without the HAG result were compared. Eighty-one (26.2 percent) patients had HAG and 11 (3.6 percent) also had ultrasound (USG) features of liver metastasis. Interval between diagnosis and treatment were significantly different between USG and HAG positive groups (Mann Whitney U test, P < 0.05). Seventeen (5.5 percent) of the 309 patients died of the disease and 7 (41.2 percent) of them had liver metastasis. Three (27.3 percent) of the 11 patients who had USG features of liver metastasis died of the disease; mortality is significantly higher than those without USG features of metastasis (Chi-square test, P < 0.05). Patients classified as medium to high risk with or without taking HAG as a feature of liver metastasis were associated with higher mortality and recurrent rate (Chi-square test, P < 0.05). On the other hand, the chance of drug resistance was higher in the medium to high risk group after reclassifying all HAG positive patients as negative for liver metastasis (Chi-square test, P < 0.05). HAG evidence of liver metastasis did not correlate with patient mortality. HAG was not an appropriate investigation in the management of GTT. PMID- 16377587 TI - Cell permeable iron chelators as potential cancer chemotherapeutic agents. AB - Iron is an essential micronutrient for the growth and function of all cells. It is, therefore, an attractive target for chemotherapeutic compounds. Numerous studies in vitro and in vivo provide evidence that iron chelators may be effective antitumor agents. Lipophilic iron chelators that are readily cell permeable and can bind intracellular iron stores may selectively kill cancer cells without damaging normal cells. In this review we discuss the role of iron in cellular processes and how these processes differ between normal and neoplastic cells. We also review the effects on normal and cancer cell growth of several lipophilic iron chelators. PMID- 16377588 TI - Developments in the management of extremity soft tissue sarcomas. AB - Soft tissue sarcomas, a rare and extremely low prevalence tumor, has no uniform treatment recommendation because there have been few randomized prospective trials to guide management. Recently, there have been surgical, medical, biologic, and genetic advances toward improved treatment. A multidisciplinary approach at specialized treatment centers should be sought because the skill set necessary to implement these advances can be provided and outcomes improved. Centralized care will be crucial to improving survival and functional outcome because it will allow for patients to be effectively entered into prospective randomized trials and create regional and national tissue banks to generate meaningful data. PMID- 16377589 TI - Pesticides and human cancers. AB - The potential for human carcinogenicity of almost all pesticides currently on the market has been poorly evaluated and is inadequately understood. Generating mechanistic data in both animal studies and epidemiology will play an increasingly important role in the future. Improved exposure assessment, in large prospective studies that generate reliable exposure-response data that focus on individual pesticide exposures are needed. One of the greatest opportunities to make more rapid progress will be to foster more multi-disciplinary collaborations between toxicologists and epidemiologists. Collaborations on molecular epidemiology investigations offers such opportunities to both toxicologists and epidemiologists that were not possible even a decade ago. PMID- 16377590 TI - Anti-angiogenic treatment of gastrointestinal malignancies. AB - The scientific rationale to block angiogenesis as a treatment strategy for human cancer has been developed over the last 30 years, but is only now entering the clinical arena. Preclinical studies have demonstrated the importance of the vascular endothelial growth factor (VEGF) pathways in both physiologic and pathologic angiogenesis, and have led to the development of approaches to block its role in tumor angiogenesis. Bevacizumab is an antibody to VEGF and has been shown to prolong survival when given with chemotherapy in the treatment of metastatic colorectal cancer (CRC). Although this is the first anti-angiogenic treatment to be approved for the treatment of human epithelial malignancy, a number of other approaches currently are in development. Soluble chimeric receptors to sequester serum VEGF and monoclonal antibodies against VEGF receptors have both shown considerable promise in the laboratory and are being brought into clinical investigation. A number of small-molecule tyrosine kinase inhibitors that have activity against VEGF receptors also are in clinical trials. Although these novel treatments are being pioneered in CRC, anti-angiogenic approaches also are being tested in the treatment of other gastrointestinal malignancies. Anti-VEGF therapy has shown promise in such traditionally resistant tumors as pancreatic cancer and hepatocellular carcinoma. This review will examine the preclinical foundation and then focus on the clinical studies of anti VEGF therapy in gastrointestinal cancers. PMID- 16377591 TI - A multidimensional model for understanding cancer pain. AB - The experience of pain pervades the physical and psychosocial domains of a patient's existence. It has a concrete underpinning in the form of an injury or disease process, yet subjective responses to pain sensations are psychosocial processes that influence the experience of pain and the capacity to cope with it. Anticipation of pain is one of the key fears associated with cancer, and uncontrolled pain strips away morale and quality of life. The interacting biopsychosocial dimensions are important areas for consideration in the comprehensive, skillful approach to assessment and treatment of cancer pain. This article addresses the interactive relationships between pain and mood using the biopsychosocial model as a heuristic for assessment. PMID- 16377592 TI - Reflections on ethical concerns arising from the incorporation of results of randomized trials of antineoplastic therapy into routine clinical practice. AB - There has been little discussion in the medical literature regarding the process whereby physicians elect to accept or reject the incorporation of results of randomized trials into their routine clinical practice. This decision-making process has the potential to result in serious ethical dilemmas. PMID- 16377593 TI - A Porsche for patient accrual. AB - A $1,000 lottery ticket for a Porsche Boxster automobile was offered to the investigator who accrued the most patients to a Southwest Oncology Group prostate cancer protocol. This was done with the admirable intention of increasing patient accrual and improving the outlook for men with high risk prostate cancer. However, the offer of this prize also makes the statement that it is permissible to reward a doctor with an expensive automobile for putting patients on an oncology research protocol. Awarding an expensive prize for patient accrual risks eroding public confidence by creating the perception that clinical investigators, swayed by the allure of an expensive automobile, were motivated by material self interest rather than the welfare of their patients and the advancement of medical science. I suspect that if the practice of expensive rewards for patient accrual became widespread the cumulative damage would ultimately outweigh the benefits. PMID- 16377595 TI - Editorial. PMID- 16377597 TI - The National Cancer Institute's cancer information service: a new generation of service and research to the nation. AB - The National Cancer Institute (NCI), the nation's leading agency for cancer information and research, mandated by the U.S. Congress to provide accurate, up to-date information about cancer to all segments of the U.S. population, established the Cancer Information Service (CIS) on July 1, 1975. Using a two pronged approach, the telephone information service and education programs for the public and health professionals, the CIS was designed to maximize its reach. In 1982, the CIS implemented three tools: the Call Record Form to record data about each call to the service, a national user survey, and a system of national test calls. These resulted in a rich data source and an infrastructure that allowed collaborative health communications research with the CIS to emerge later in the decade. As the CIS embarks on a new generation of service to the nation, it is now characterized by three vital components that advance the NCI's overall mission of cancer prevention and control: a Partnership Program, Multichannel Contact Centers, and a Health Communications Research Program. PMID- 16377598 TI - Cancer patients' information needs across the cancer care continuum: evidence from the cancer information service. AB - This study examines the information needs of cancer patients who contacted the National Cancer Institute's (NCI's) Cancer Information Service (CIS) via a toll free telephone number. Records from 19,030 calls received from cancer patients between September 2002 and August 2003 were analyzed to determine differences in subjects of interaction (main topics of inquiry and discussion) for subgroups of patients based on demographic characteristics and stage along the cancer care continuum (pretreatment, in-treatment, post-treatment, recurrence). Females were more likely than males to inquire about cancer screening/diagnosis, support services, psychosocial issues, and general cancer site information, but they were less likely to seek specific cancer treatment information. Older patients were more likely than younger patients to seek specific treatment information, but they were less interested in support services, psychosocial issues, and prevention/risk factors. Compared with White callers, Hispanics and most minorities were more likely to seek support service information, and African Americans were more likely to have questions related to psychosocial issues. Compared with patients in treatment, patients in recurrence were more likely to seek specific treatment information; patients not in treatment were more likely to seek medical referral information; and patients in post-treatment were more likely to seek screening/diagnosis and prevention/risk factor information. Findings will help the CIS and other cancer-focused organizations address the distinct information needs of different subsets of cancer patients. PMID- 16377599 TI - Exploring e-Health usage and interest among cancer information service users: the need for personalized interactions and multiple channels remains. AB - Since searching for health information is among the most popular uses of the Internet, we analyzed a survey of 6,019 callers to the National Cancer Institute's (NCI's) Cancer Information Service (CIS) to assess Internet usage and interest in technologies to access health and cancer information. Findings suggest that about 40% of CIS callers used the Internet to obtain cancer information and, of these, only about 20% found all the information they sought. Nearly 33% of Internet users called the CIS to discuss information found on the Internet; most (>90%) reported that the CIS was helpful. Those who sought cancer information on the Internet were more likely to call the CIS about this information if they found all or most of the information they were seeking, compared with those who found some or little of the information. New communication services endorsed by most CIS callers included e-mails from an information specialist and telephone support from the CIS while on the Internet. The survey results indicate the importance of multiple access points, both traditional and technology based, and that there is still a need for more traditional, personalized forms of health communication. A crucial question is how best to harness and integrate these new technologies within the current generation of mediated health information systems. PMID- 16377600 TI - Information-seeking behavior of minority breast cancer patients: an exploratory study. AB - Few population-based studies have been conducted with minority breast cancer patients in the United States to understand their information-seeking behaviors. We examined the resources minority breast cancer patients used and the extent to which the Internet was being utilized by Hispanic and African American breast cancer patients. A random sample of 388 Hispanic and African American breast cancer female patients was obtained from the Sacramento Cancer Surveillance Program in California. Various survey and data collection tools were researched and adapted to create the 44-item questionnaire. Subjects were asked to participate in a telephone interview. Of the 388 cases, 74% (n = 287) consented to participate in our study. Sixty-three percent (n = 181) received cancer related information at the time of their diagnosis; 58% reported that a health care professional provided them with this information. Only 17% of study respondents had ever used the Internet. The top 3 sources for cancer information were books, brochures, and pamphlets (98%); doctor or other health professionals (97%); and spouse or partner, family members, friends, or all of these (62%). None of the respondents indicated accessing the National Cancer Institute's cancer.gov website. Minority breast cancer patients were using the Internet for cancer-related information at a very low rate. PMID- 16377601 TI - Multiple tailored messages are effective in increasing fruit and vegetable consumption among callers to the Cancer Information Service. AB - Results are reported from a large (n = 3,402) four-group randomized trial to increase fruit and vegetable consumption among callers to the National Cancer Institute's (NCI's) Cancer Information Service (CIS) using tailored print materials. Following a baseline telephone interview, which included a brief educational message (BEM), participants were assigned randomly within CIS offices to one of four groups: single untailored (SU) group-one untailored set of materials; single tailored (ST) group-one tailored booklet; multiple tailored (MT) group-four tailored materials; and multiple retailored (MRT) group-four tailored materials with retailoring based on new information obtained at 5 months follow-up. Follow-up telephone interviews were conducted at 5 (n = 2,233) and 12 months (n = 1,927) after baseline. The main outcome measure was self-reported fruit and vegetable consumption using a seven-item food frequency questionnaire. At 12 months follow-up, there was a significant linear trend across groups of 0.21 servings (p = 0.0002). Specific nested hypotheses then were tested and revealed significant mean serving differences between SU (5.07) vs. MT (5.64) (p = 0.002) and SU vs. MRT (5.71; p < 0.001). Although the mean for ST (5.40) was greater than that for SU (5.07), the difference was not statistically significant (p = 0.07), and no difference was found between MT vs. MRT (p = 0.69). A higher proportion of recipients of tailored materials reported reading all of the materials and believing that they were written especially for them. No differences by experimental condition were found for the perceived usefulness or motivational impact of the print materials. In this trial, MT print materials were more effective at increasing fruit and vegetable (FV) consumption than were SU materials. The intervention mechanisms responsible for this effect merit further research. Retailoring did not produce a significant difference when compared with longitudinal baseline tailoring. PMID- 16377602 TI - The efficacy of tailored print materials in promoting colorectal cancer screening: results from a randomized trial involving callers to the National Cancer Institute's Cancer Information Service. AB - In this large randomized trial among callers to the Cancer Information Service (CIS), tailored print materials were tested for efficacy in promoting colorectal cancer (CRC) screening (fecal occult blood test [FOBT], flexible sigmoidoscopy, or colonoscopy). All participants completed baseline interviews at the end of their usual service calls to the CIS, as well as short-term (6-month) and longer term (14-month) telephone follow-up interviews. The study sample (n = 4,014) was restricted to English-speaking CIS callers 50 + years of age, who would be eligible for CRC screening at 14 months follow-up and did not call the CIS about CRC or CRC screening. Four experimental conditions were compared: a single untailored (SU) mailout of print material (the control condition); a single tailored (ST) mailout of print material; four (multiple) tailored (MT) mailouts of print materials spanning 12 months, all of which were tailored to information obtained at baseline; and four (multiple) retailored (MRT) mailouts also spanning 12 months, with retailoring of the print materials (mailouts 2, 3, and 4) based on updated information obtained from the 6-month follow-up interviews. Consistent with the main hypothesis of this trial, a significant linear trend across the SU, ST, MT, and MRT groups was found at 14 months (42%, 44%, 51%, and 48%, respectively, p = 0.05). Only for MT was there a significant difference compared with SU (p = 0.03) for the sample as a whole, while no differences were found for MT vs. MRT at 14 months. Significant moderator effects in the predicted direction were found among females, younger participants, and among those with a history of CRC screening, all of which involved the SU vs. MT MRT comparisons. Only among younger participants (ages 50-59) was there a difference between SU vs. ST at 14 months. Given these results, we conclude from this trial the following: (1) the MRT intervention failed to show added benefit beyond the MT intervention, (2) the significant intervention effects involving the MT and MRT conditions can be explained by tailoring and/or the longitudinal nature of both interventions, and (3) the most compelling evidence in support of tailoring was found for the ST condition among younger participants, where a significant need for interventions exists at the national level. Directions for future research are discussed in light of the results summarized above. PMID- 16377603 TI - A randomized controlled trial of multiple tailored messages for smoking cessation among callers to the cancer information service. AB - Self-help materials computer-tailored to the specific needs of smokers have shown promise as a high-reach, low-cost intervention for smoking cessation. Adding tailored cessation materials to telephone-based cessation counseling may be a way of generating greater efficacy in promoting and maintaining cessation. The objective of this study is to assess the efficacy of adding different types of behavioral smoking cessation materials to brief telephone-based cessation counseling.A total of 1,978 smokers calling the National Cancer Institute's (NCI's) Cancer Information Service (CIS) for help in quitting smoking initially received brief cognitive-behavioral cessation counseling from a CIS information specialist. Following a baseline interview administered by the information specialist, subjects were randomly assigned to one of four conditions, each delivered by U.S. mail: a single, untailored smoking cessation guide (SU); a single, tailored smoking cessation guide (ST); a series of four (multiple) printed materials tailored only to baseline data (MT); and a series of four (multiple) printed materials tailored to baseline as well as retailored using 5 month interim progress data (MRT). The primary outcome measure was 7-day point prevalence abstinence rates assessed using a computer-assisted telephone interview (CATI) at 12-month follow-up.At 12-month follow-up, using intent-to treat, imputed, and per-protocol analyses, no differences were found among the four experimental conditions (linear trend), or when the ST, MT, and MRT groups were compared with the control (SU) group. Participants in the two multiple message group conditions combined (MT + MRT), however, had significantly higher abstinence rates than participants in the two single message group conditions combined (SU + ST). Moreover, among subjects who reported quitting at the 5-month follow-up, participants receiving the MRT materials reported higher abstinence rates at 12 months than the other three groups combined (SU + ST + MT). The results of this study support the effectiveness, over and above a single telecounseling interaction, of multiple tailored print material contacts on cessation. These effects, however may be due to tailoring, or the longitudinal nature of the two multiple tailored conditions, or both. The strongest evidence for tailoring occurred in the MRT condition for relapse prevention, suggesting that print materials tailored to interim progress may be especially effective in this context. The qualities of specific psychosocial and communication elements in tailored materials should receive attention in future research. PMID- 16377604 TI - Facilitating informed decision making about breast cancer risk and genetic counseling among women calling the NCI's Cancer Information Service. AB - Despite increased interest among the public in breast cancer genetic risk and genetic testing, there are limited services to help women make informed decisions about genetic testing. This study, conducted with female callers (N = 279) to the National Cancer Institute's (NCI's) Atlantic Region Cancer Information Service (CIS), developed and evaluated a theory-based, educational intervention designed to increase callers' understanding of the following: (a) the kinds of information required to determine inherited risk; (b) their own personal family history of cancer; and (c) the benefits and limitations of genetic testing. Callers requesting information about breast/ovarian cancer risk, risk assessment services, and genetic testing were randomized to either: (1) standard care or (2) an educational intervention. Results show that the educational intervention reduced intention to obtain genetic testing among women at average risk and increased intention among high-risk women at 6 months. In addition, high monitors, who typically attend to and seek information, demonstrated greater increases in knowledge and perceived risk over the 6-month interval than low monitors, who typically are distracted from information. These findings suggest that theoretically designed interventions can be effective in helping women understand their cancer risk and appropriate risk assessment options and can be implemented successfully within a service program like the CIS. PMID- 16377605 TI - Motivating cancer prevention and early detection behaviors using psychologically tailored messages. AB - Tailoring health messages to make them salient to recipients is a strategy to motivate cancer prevention and early detection behaviors. Various tactics can be used to tailor health materials; our approach involves tailoring messages to individual differences in the psychological processes by which people understand health information. To summarize our tailoring approach, we review findings from six field experiments (four published, two pending publication) conducted in collaboration with the National Cancer Institute's (NCI's) Cancer Information Service (CIS) examining the utility of psychologically tailored messages. Messages were tailored to individual variability in the following: (a) need for cognition, (b) monitor/blunting coping style, (c) health locus of control, and (d) regulatory focus. Collectively, the findings suggest that, as hypothesized, messages congruent with the recipients' psychological style of health information processing (i.e., matched messages) are more persuasive in promoting screening mammography and fruit and vegetable consumption than mismatched messages. This line of research provides evidence for the utility of psychological tailoring as a health communication strategy and direction for developing effective health messages in a variety of settings. PMID- 16377606 TI - Raising awareness of on-line cancer information: helping providers empower patients. AB - While the digital divide remains a special problem in health care, providers' reluctance to refer patients to the Internet is an equally important problem. The Bridging the Digital Divide Project: Your Access to Cancer Information was designed with two target audiences in mind-consumers and health care providers. A total of 256 providers from varied health care settings enrolled in workshops over a 10-month period (2001-2002). Results suggest differences in awareness and use of on-line cancer information among providers and confirm that many providers need to become more comfortable with referring patients to on-line information. At completion of the workshops, all participants reported an increased awareness of cancer information websites, increased confidence in making judgments about the reliability and appropriateness of the sites, an increased willingness to discuss Internet information sources with patients and their family members, and an increased awareness of and intention to use the CIS. Providers from the community hospital were more likely than other groups to report that what they had learned about the Internet was helpful and that their comfort level using the Internet had increased. Partnerships between providers and the CIS may help to further increase this comfort level, ultimately benefiting cancer patients. PMID- 16377607 TI - Reducing the digital divide for low-income women with breast cancer: a feasibility study of a population-based intervention. AB - A fundamental challenge to helping underserved women and their families cope with breast cancer is providing them with easily accessible, reliable health care information and support. This is especially true for low-income families living in rural areas where resources are few and frequently distant as well as low income families in urban areas where access to information and support can be complex and overwhelming. The Internet is one mechanism that has tremendous potential to help these families cope with breast cancer. This article describes a feasibility test of the potential for the National Cancer Institute's (NCI's) Cancer Information Service (CIS) to provide access to an Internet-based system that has been shown to improve quality of life for underserved breast cancer patients. The test was conducted in rural Wisconsin (low socioeconomic status [SES] Caucasian women) and in Detroit, Michigan (low SES African American women), and compares the effectiveness of several different dissemination strategies. Using these results we propose a model for how CIS telephone and partnership program services could efficiently disseminate such information and support systems. In doing so we believe that important steps can be taken to close the digital divide that separates low-income families from the resources they need to effectively face cancer. This is the first of two articles coming from this study. A companion article reports on an evaluation of the use and impact of this system on the women who were given access to it. PMID- 16377608 TI - Use and Impact of eHealth System by Low-income Women With Breast Cancer. AB - This article is the second of a two-part series reporting on a population-based study intended to use an eHealth system to examine the feasibility of reaching underserved women with breast cancer (Gustafson, McTavish et al., Reducing the digital divide for low-income women with breast cancer, 2004; Madison Center for Health Systems Research and Analysis, University of Wisconsin; Comprehensive Health Enhancement Support System [CHESS]) and determine how they use the system and what impact it had on them. Participants included women recently diagnosed with breast cancer whose income was at or below 250% of poverty level and were living in rural Wisconsin (n = 144; all Caucasian) or Detroit (n = 85; all African American). Because this was a population-based study all 229 participants received CHESS. A comparison group of patients (n = 51) with similar demographics was drawn from a separate recently completed randomized clinical trial. Use rates (e.g., frequency and length of use as well as type of use) as well as impact on several dimensions of quality of life and participation in health care are reported. Low-income subjects in this study logged on and spent more time on CHESS than more affluent women in a previous study. Urban African Americans used information and analysis services more and communication services less than rural Caucasians. When all low-income women from this study are combined and compared with a low-income control group from another study, the CHESS group was superior to that control group in 4 of 8 outcome variables at both statistically and practically significant levels (social support, negative emotions, participation in health care, and information competence). When African Americans and Caucasians are separated the control group's sample size becomes 30 and 21 thus reducing power. Statistical significance is retained, however, in all four outcomes for Caucasians and in two of four for African Americans. Practical significance is retained for all four outcomes. We conclude that an eHealth system like CHESS will be used extensively and have a positive impact on low income women with breast cancer. PMID- 16377609 TI - A case study in dissemination: lessons learned from a pilot study involving the National Cancer Institute's Cancer Information Service. AB - The Cancer Information Service Research Consortium (CISRC) was funded by the National Cancer Institute to disseminate as a pilot study a longitudinally tailored print intervention to promote the 5 A Day for Better Health program among callers to the National Cancer Institute's Cancer Information Service (CIS). Using a one-group (intervention-only) study design, 1,022 eligible CIS callers were enrolled to receive the intervention consisting of four mailings of tailored print materials over a 3-month period. Program evaluation focused on process and implementation evaluation, including adherence to the baseline interviews by CIS information specialists based on live-call monitoring (n = 55 eligible callers), and the timeliness of the intervention mailouts (4,088 scheduled mailouts). Adherence to the baseline interviews by CIS information specialists was extremely high, exceeding 90% for all indicators of quality control. Of the 4,088 intervention mailings, 75% occurred on or before the target date, while 95% occurred within 21 days of the target date. All delays in the scheduled mailouts occurred in the first mailing, due to changes made in the production process (batch printing of all tailored print materials at baseline). This change required additional system upgrades and more intensive and time consuming quality control than originally anticipated, which was exacerbated by the faster-than-expected accrual of eligible participants. Based on this pilot study, the CIS is now positioned for widespread dissemination of the 5 A Day tailored print intervention. Several key lessons learned are also identified to facilitate the transition from research to dissemination. PMID- 16377610 TI - The CIS model for collaborative research in health communications: a brief retrospective from the current generation of research. AB - The Cancer Information Service (CIS) of the National Cancer Institute (NCI) is the premiere organization for providing cancer information to the nation. The CIS provides a stellar example of how a service organization dedicated to health communications also can serve as a laboratory for research. This journey by the CIS into health communication research is described briefly, along with the current generation of research summarized in this issue of the Journal of Health Communication (JHC). The CIS model for collaborative research is presented as an exemplar that other service organizations might embrace as a strategic tool for quality improvement in health communications. PMID- 16377611 TI - ChroCoLoc: an application for calculating the probability of co-localization of microarray gene expression. AB - With the production of whole genome microarray chips the ability arises to investigate whether the regulation of particular groups of genes may be influenced by their chromosomal localization. Chromosome Co-Localization probability calculator (ChroCoLoc) is a publicly available web-based tool for the analysis of co-localization of co-expressed genes identified by microarray experiments. AVAILABILITY: http://www.ebi.ac.uk/expressionprofiler/ PMID- 16377612 TI - GenomeDiagram: a python package for the visualization of large-scale genomic data. AB - We present GenomeDiagram, a flexible, open-source Python module for the visualization of large-scale genomic, comparative genomic and other data with reference to a single chromosome or other biological sequence. GenomeDiagram may be used to generate publication-quality vector graphics, rastered images and in line streamed graphics for webpages. The package integrates with datatypes from the BioPython project, and is available for Windows, Linux and Mac OS X systems. AVAILABILITY: GenomeDiagram is freely available as source code (under GNU Public License) at http://bioinf.scri.ac.uk/lp/programs.html, and requires Python 2.3 or higher, and recent versions of the ReportLab and BioPython packages. SUPPLEMENTARY INFORMATION: A user manual, example code and images are available at http://bioinf.scri.ac.uk/lp/programs.html. PMID- 16377613 TI - Improving missing value estimation in microarray data with gene ontology. AB - MOTIVATION: Gene expression microarray experiments produce datasets with frequent missing expression values. Accurate estimation of missing values is an important prerequisite for efficient data analysis as many statistical and machine learning techniques either require a complete dataset or their results are significantly dependent on the quality of such estimates. A limitation of the existing estimation methods for microarray data is that they use no external information but the estimation is based solely on the expression data. We hypothesized that utilizing a priori information on functional similarities available from public databases facilitates the missing value estimation. RESULTS: We investigated whether semantic similarity originating from gene ontology (GO) annotations could improve the selection of relevant genes for missing value estimation. The relative contribution of each information source was automatically estimated from the data using an adaptive weight selection procedure. Our experimental results in yeast cDNA microarray datasets indicated that by considering GO information in the k-nearest neighbor algorithm we can enhance its performance considerably, especially when the number of experimental conditions is small and the percentage of missing values is high. The increase of performance was less evident with a more sophisticated estimation method. We conclude that even a small proportion of annotated genes can provide improvements in data quality significant for the eventual interpretation of the microarray experiments. AVAILABILITY: Java and Matlab codes are available on request from the authors. SUPPLEMENTARY MATERIAL: Available online at http://users.utu.fi/jotatu/GOImpute.html. PMID- 16377614 TI - The structure of the interleukin-15 alpha receptor and its implications for ligand binding. AB - Interleukin (IL)-15 is a member of the small four alpha-helix bundle family of cytokines. IL-15 was discovered by its ability to mimic IL-2-mediated T-cell proliferation. Both cytokines share the beta and gamma receptor chains of the IL 2 receptor for signal transduction. However, in addition, they target specific alpha chain receptors IL-15Ralpha and IL-2Ralpha, respectively. The exceptionally high affinity binding of IL-15 to IL-15Ralpha is mediated by its sushi domain. Here we present the solution structure of the IL-15Ralpha sushi domain solved by NMR spectroscopy and a model of its complex with IL-15. The model shows that, rather than the familiar hydrophobic forces dominating the interaction interface between cytokines and their cognate receptors, the interaction between the IL-15 and IL-15Ralpha complex involves a large network of ionic interactions. This type of interaction explains the exceptionally high affinity of the IL-15.IL-15Ralpha complex, which is essential for the biological effects of this important cytokine and which is not observed in other cytokine/cytokine receptor complexes. PMID- 16377615 TI - Inhibition of RecA protein function by the RdgC protein from Escherichia coli. AB - The Escherichia coli RdgC protein is a potential negative regulator of RecA function. RdgC inhibits RecA protein-promoted DNA strand exchange, ATPase activity, and RecA-dependent LexA cleavage. The primary mechanism of RdgC inhibition appears to involve a simple competition for DNA binding sites, especially on duplex DNA. The capacity of RecA to compete with RdgC is improved by the DinI protein. RdgC protein can inhibit DNA strand exchange catalyzed by RecA nucleoprotein filaments formed on single-stranded DNA by binding to the homologous duplex DNA and thereby blocking access to that DNA by the RecA nucleoprotein filaments. RdgC protein binds to single-stranded and double stranded DNA, and the protein can be visualized on DNA using electron microscopy. RdgC protein exists in solution as a mixture of oligomeric states in equilibrium, most likely as monomers, dimers, and tetramers. This concentration-dependent change of state appears to affect its mode of binding to DNA and its capacity to inhibit RecA. The various species differ in their capacity to inhibit RecA function. PMID- 16377616 TI - Membrane insertion of Escherichia coli alpha-hemolysin is independent from membrane lysis. AB - Escherichia coli alpha-hemolysin (HlyA) is a protein exotoxin that binds and lyses eukaryotic cell and model membranes in the presence of calcium. Previous studies have been able to distinguish between reversible toxin binding to the membrane and irreversible insertion into the lipid matrix. Membrane lysis occurs as the combined effect of protein insertion plus a transient perturbation of the membrane bilayer structure. In the past, insertion and bilayer perturbation have not been experimentally dissected. This has now been achieved by studying HlyA penetration into lipid monolayers at the air-water interface, in which three dimensional effects (of the kind required to break down the bilayer permeability barrier) cannot occur. The study of native HlyA, together with the nonlytic precursor pro-HlyA, and of different mutants demonstrates that although some nonlytic variants (e.g. pro-HlyA) exhibit very low levels of insertion, others (e.g. the nonlytic mutant HlyA H859N) insert even more strongly than the lytic wild type. These results show that insertion does not necessarily lead to membrane lysis, i.e. that insertion and lysis are not "coupled" phenomena. Millimolar levels of Ca(2+), which are essential for the lytic activity, cause an extra degree of insertion but only in the case of the lytic forms of HlyA. PMID- 16377617 TI - A reassessment of the FNR regulon and transcriptomic analysis of the effects of nitrate, nitrite, NarXL, and NarQP as Escherichia coli K12 adapts from aerobic to anaerobic growth. AB - The transcription factor FNR, the regulator of fumarate and nitrate reduction, regulates major changes as Escherichia coli adapts from aerobic to anaerobic growth. In an anaerobic glycerol/trimethylamine N-oxide/fumarate medium, the fnr mutant grew as well as the parental strain, E. coli K12 MG1655, enabling us to reveal the response to oxygen, nitrate, and nitrite in the absence of glucose repression or artifacts because of variations in growth rate. Hence, many of the discrepancies between previous microarray studies of the E. coli FNR regulon were resolved. The current microarray data confirmed 31 of the previously characterized FNR-regulated operons. Forty four operons not previously known to be included in the FNR regulon were activated by FNR, and a further 28 operons appeared to be repressed. For each of these operons, a match to the consensus FNR binding site sequence was identified. The FNR regulon therefore minimally includes at least 103, and possibly as many as 115, operons. Comparison of transcripts in the parental strain and a narXL deletion mutant revealed that transcription of 51 operons is activated, directly or indirectly, by NarL, and a further 41 operons are repressed. The narP gene was also deleted from the narXL mutant to reveal the extent of regulation by phosphorylated NarP. Fourteen promoters were more active in the narP+ strain than in the mutant, and a further 37 were strongly repressed. This is the first report that NarP might function as a global repressor as well as a transcription activator. The data also revealed possible new defense mechanisms against reactive nitrogen species. PMID- 16377618 TI - The endonuclease domain of bacteriophage terminases belongs to the resolvase/integrase/ribonuclease H superfamily: a bioinformatics analysis validated by a functional study on bacteriophage T5. AB - Bacteriophage terminases are essential molecular motors involved in the encapsidation of viral DNA. They are hetero-multimers whose large subunit encodes both ATPase and endonuclease activities. Although the ATPase domain is well characterized from sequence and functional analysis, the C-terminal region remains poorly defined. We describe sequence-structure comparisons of the endonuclease region of various bacteriophages that revealed new sequence similarities shared by this region and the Holliday junction resolvase RuvC and to a lesser extent the HIV integrase and the ribonuclease H. Extensive sequence comparison and motif refinement led to a common signature of terminases and resolvases with three conserved acidic residues engaged in catalytic activity. Sequence analyses were validated by in vivo and in vitro functional assays showing that the nuclease activity of the endonuclease domain of bacteriophage T5 terminase was abolished by mutation of any of the three predicted catalytic aspartates. Overall, these data suggest that the endonuclease domains of terminases operate autonomously and that they adopt a fold similar to that of resolvases and share the same divalent cation-dependent enzymatic mechanism. PMID- 16377619 TI - Histone H1 phosphorylation occurs site-specifically during interphase and mitosis: identification of a novel phosphorylation site on histone H1. AB - H1 histones, isolated from logarithmically growing and mitotically enriched human lymphoblastic T-cells (CCRF-CEM), were fractionated by reversed phase and hydrophilic interaction liquid chromatography, subjected to enzymatic digestion, and analyzed by amino acid sequencing and mass spectrometry. During interphase the four H1 subtypes present in these cells differ in their maximum phosphorylation levels: histone H1.5 is tri-, H1.4 di-, and H1.3 and H1.2, only monophosphorylated. The phosphorylation is site-specific and occurs exclusively on serine residues of SP(K/A)K motifs. The phosphorylation sites of histone H1.5 from mitotically enriched cells were also examined. In contrast to the situation in interphase, at mitosis there were additional phosphorylations, exclusively at threonine residues. Whereas the tetraphosphorylated H1.5 arises from the triphosphosphorylated form by phosphorylation of one of two TPKK motifs in the C terminal domain, namely Thr137 and Thr154, the pentaphosphorylated H1.5 was the result of phosphorylation of one of the tetraphosphorylated forms at a novel nonconsensus motif at Thr10 in the N-terminal tail. Despite the fact that histone H1.5 has five (S/T)P(K/A)K motifs, all of these motifs were never found to be phosphorylated simultaneously. Our data suggest that phosphorylation of human H1 variants occurs nonrandomly during both interphase and mitosis and that distinct serine- or threonine-specific kinases are involved in different cell cycle phases. The order of increased phosphorylation and the position of modification might be necessary for regulated chromatin decondensation, thus facilitating processes of replication and transcription as well as of mitotic chromosome condensation. PMID- 16377620 TI - Apoptotic cells, at all stages of the death process, trigger characteristic signaling events that are divergent from and dominant over those triggered by necrotic cells: Implications for the delayed clearance model of autoimmunity. AB - Current models of autoimmunity suggest that delayed clearance of apoptotic cells leads to the presentation of apoptotic antigens in the context of inflammatory signals, with resultant autoimmunity. These models implicitly assume that, in contrast to early apoptotic cells (that retain membrane integrity), late apoptotic cells (with compromised membranes) act like necrotic cells (which also lack intact membranes), possibly because of the release of proinflammatory intracellular contents. We showed previously that early apoptotic and necrotic cells induce distinct mitogen-activated protein kinase modules in macrophages with which they interact. Exposure to apoptotic cells led to nearly complete inhibition of both basal and macrophage colony-stimulating factor-induced ERK1/2 by macrophages. In contrast, necrotic cells induced ERK1/2. We show here that apoptotic cells also strongly induced both c-Jun N-terminal kinase and p38, whereas necrotic cells had no detectable effect on c-Jun N-terminal kinase and p38. We also compared the signaling events induced in macrophages by exposure to early apoptotic cells, late apoptotic cells, and necrotic cells. The signaling events induced by late apoptotic cells were identical to and just as potent as those induced by early apoptotic cells. Thus, apoptotic cells are functionally equivalent throughout the cell death process, irrespective of membrane integrity. Moreover, the effects of both early and late apoptotic cells on signaling were dominant over those of necrotic cells. These data show that apoptotic cells do not become proinflammatory upon the loss of membrane integrity and are inconsistent with the notion that delayed clearance alone can lead to autoimmunity. PMID- 16377621 TI - The role of human HtrA1 in arthritic disease. AB - Human HtrA1 belongs to a widely conserved family of serine proteases involved in various aspects of protein quality control and cell fate. Although HtrA1 has been implicated in the pathology of several diseases, its precise biological functions remain to be established. Through identification of potential HtrA1 targets, studies presented herein propose that within the context of arthritis pathology HtrA1 contributes to cartilage degradation. Elevated synovial HtrA1 levels were detected in fluids obtained from rheumatoid and osteoarthritis patients, with synovial fibroblasts identified as a major source of secreted HtrA1. Mass spectrometry analysis of potential HtrA1 substrates within synovial fluids identified fibronectin as a candidate target, and treatment of fibronectin with recombinant HtrA1 led to the generation of fibronectin-degradation products that may be involved in cartilage catabolism. Consistently, treatment of synovial fibroblasts with HtrA1 or HtrA1-generated fibronectin fragments resulted in the specific induction of matrix metalloprotease 1 and matrix metalloprotease 3 expression, suggesting that HtrA1 contributes to the destruction of extracellular matrix through both direct and indirect mechanisms. PMID- 16377622 TI - Expression and characterization of the peptidase domain of Streptococcus pneumoniae ComA, a bifunctional ATP-binding cassette transporter involved in quorum sensing pathway. AB - ComA, a member of the bacteriocin ATP-binding cassette transporters, is postulated to be responsible for both the processing of the propeptide ComC and secretion of the mature competence-stimulating peptide, which regulates the competence and subsequent genetic transformation in Streptococcus pneumoniae. A recombinant N-terminal peptidase domain of ComA, designated PEP, was expressed as a soluble protein in Escherichia coli, purified to homogeneity, and characterized. Gel-filtration analysis revealed that PEP functions as a monomer. The purified PEP exhibited an efficient proteolytic activity for the substrate ComC, which was cleaved after the double glycine site. The stability of PEP was examined by circular dichroism analyses. A convenient method for analyzing the proteolytic activity of PEP was developed, and the kinetic parameters for ComC were determined (k(cat) = 1.5 +/- 0.083 min(-1) and Km = 62 +/- 9.0 microM). Replacements of Cys17 of PEP with Ser or Ala and His96 with Ala resulted in complete loss of activity, indicating that both Cys17 and His96 are essential for the catalysis. Together with information from a protease data base, the N terminal domain of ComA was concluded to belong to the same clan as the papain like cysteine proteases. Mutant substrates, in which each of the double glycines was replaced with Ala, were cleaved very poorly by PEP. The mechanism of this strict substrate specificity is discussed on the basis of the sequence alignment with other cysteine proteases. PMID- 16377623 TI - Desmoglein endocytosis and desmosome disassembly are coordinated responses to pemphigus autoantibodies. AB - Desmosomes are adhesive intercellular junctions prominent in the skin and heart. Loss of desmosome function is associated with severe congenital and acquired disorders characterized by tissue fragility. Pemphigus vulgaris (PV) is an autoimmune disorder in which antibodies are directed against the desmosomal adhesion molecule Dsg3, resulting in severe mucosal erosions and epidermal blistering. To define the mechanisms by which Dsg3 autoantibodies disrupt keratinocyte adhesion, the fate of PV IgG and various desmosomal components was monitored in primary human keratinocytes exposed to PV patient IgG. PV IgG initially bound to keratinocyte cell surfaces and colocalized with desmosomal markers. Within 6 h after PV IgG binding to Dsg3, electron microscopy revealed that desmosomes were dramatically disrupted and keratinocyte adhesion was severely compromised. Immunofluorescence analysis indicated that PV IgG and Dsg3 were rapidly internalized from the cell surface in a complex with plakoglobin but not desmoplakin. Dsg3 internalization was associated with retraction of keratin filaments from cell-cell borders. Furthermore, the internalized PV IgG-Dsg3 complex colocalized with markers for both endosomes and lysosomes, suggesting that Dsg3 was targeted for degradation. Consistent with this possibility, biotinylation experiments demonstrated that soluble Dsg3 cell surface pools were rapidly depleted followed by loss of detergent-insoluble Dsg3. These findings demonstrate that Dsg3 endocytosis, keratin filament retraction, and the loss of keratinocyte cell-cell adhesion are coordinated responses to PV IgG. PMID- 16377624 TI - Protein kinase C delta regulates Ser46 phosphorylation of p53 tumor suppressor in the apoptotic response to DNA damage. AB - The p53 tumor suppressor is activated in the cellular response to genotoxic stress. Transactivation of p53 target genes dictates cell cycle arrest and DNA repair or induction of apoptosis; however, a molecular mechanism responsible for these distinct functions remains unclear. Recent studies revealed that phosphorylation of p53 on Ser(46) was associated with induction of p53AIP1 expression, resulting in the commitment of the cell fate into apoptotic cell death. Moreover, upon exposure to genotoxic stress, p53DINP1 was expressed and recruited a kinase(s) to p53 that specifically phosphorylated Ser(46). Here, we show that the pro-apoptotic kinase, protein kinase C delta (PKCdelta), is involved in phosphorylation of p53 on Ser(46). PKCdelta-mediated phosphorylation is required for the interaction of PKCdelta with p53. The results also demonstrate that p53DINP1 associates with PKCdelta upon exposure to genotoxic agents. Consistent with these results, PKCdelta potentiates p53-dependent apoptosis by Ser(46) phosphorylation in response to genotoxic stress. These findings indicate that PKCdelta regulates p53 to induce apoptotic cell death in the cellular response to DNA damage. PMID- 16377625 TI - Substrate specificity and domain functions of extracellular heparan sulfate 6-O endosulfatases, QSulf1 and QSulf2. AB - The extracellular sulfatases (Sulfs) are an evolutionally conserved family of heparan sulfate (HS)-specific 6-O-endosulfatases. These enzymes remodel the 6-O sulfation of cell surface HS chains to promote Wnt signaling and inhibit growth factor signaling for embryonic tissue patterning and control of tumor growth. In this study we demonstrate that the avian HS endosulfatases, QSulf1 and QSulf2, exhibit the same substrate specificity toward a subset of trisulfated disaccharides internal to HS chains. Further, we show that both QSulfs associate exclusively with cell membrane and are enzymatically active on the cell surface to desulfate both cell surface and cell matrix HS. Mutagenesis studies reveal that conserved amino acid regions in the hydrophilic domains of QSulf1 and QSulf2 have multiple functions, to anchor Sulf to the cell surface, bind to HS substrates, and to mediate HS 6-O-endosulfatase enzymatic activity. Results of our current studies establish the hydrophilic domain (HD) of Sulf enzymes as an essential multifunctional domain for their unique endosulfatase activities and also demonstrate the extracellular activity of Sulfs for desulfation of cell surface and cell matrix HS in the control of extracellular signaling for embryonic development and tumor progression. PMID- 16377626 TI - Loss of PECAM-1 function impairs alveolarization. AB - The final stage of lung development in humans and rodents occurs principally after birth and involves the partitioning of the large primary saccules into smaller air spaces by the inward protrusion of septae derived from the walls of the saccules. Several observations in animal models implicate angiogenesis as critical to this process of alveolarization, but all anti-angiogenic treatments examined to date have resulted in endothelial cell (EC) death. We therefore targeted the function of platelet endothelial cell adhesion molecule, (PECAM-1), an EC surface molecule that promotes EC migration and has been implicated in in vivo angiogenesis. Administration of an anti-PECAM-1 antibody that inhibits EC migration, but not proliferation or survival in vitro, disrupted normal alveolar septation in neonatal rat pups without reducing EC content. Three-dimensional reconstruction of lungs showed that pups treated with a blocking PECAM-1 antibody had remodeling of more proximal branches resulting in large tubular airways. Subsequent studies in PECAM-1-null mice confirmed that the absence of PECAM-1 impaired murine alveolarization, without affecting EC content, proliferation, or survival. Further, cell migration was reduced in lung endothelial cells isolated from these mice. These data suggest that the loss of PECAM-1 function compromises postnatal lung development and provide evidence that inhibition of EC function, in contrast to a loss of viable EC, inhibits alveolarization. PMID- 16377627 TI - Acetobacter turbidans alpha-amino acid ester hydrolase: how a single mutation improves an antibiotic-producing enzyme. AB - The alpha-amino acid ester hydrolase (AEH) from Acetobacter turbidans is a bacterial enzyme catalyzing the hydrolysis and synthesis of beta-lactam antibiotics. The crystal structures of the native enzyme, both unliganded and in complex with the hydrolysis product D-phenylglycine are reported, as well as the structures of an inactive mutant (S205A) complexed with the substrate ampicillin, and an active site mutant (Y206A) with an increased tendency to catalyze antibiotic production rather than hydrolysis. The structure of the native enzyme shows an acyl binding pocket, in which D-phenylglycine binds, and an additional space that is large enough to accommodate the beta-lactam moiety of an antibiotic. In the S205A mutant, ampicillin binds in this pocket in a non productive manner, making extensive contacts with the side chain of Tyr(112), which also participates in oxyanion hole formation. In the Y206A mutant, the Tyr(112) side chain has moved with its hydroxyl group toward the catalytic serine. Because this changes the properties of the beta-lactam binding site, this could explain the increased beta-lactam transferase activity of this mutant. PMID- 16377628 TI - The PII signal transduction protein of Arabidopsis thaliana forms an arginine regulated complex with plastid N-acetyl glutamate kinase. AB - The PII proteins are key mediators of the cellular response to carbon and nitrogen status and are found in all domains of life. In eukaryotes, PII has only been identified in red algae and plants, and in these organisms, PII localizes to the plastid. PII proteins perform their role by assessing cellular carbon, nitrogen, and energy status and conferring this information to other proteins through protein-protein interaction. We have used affinity chromatography and mass spectrometry to identify the PII-binding proteins of Arabidopsis thaliana. The major PII-interacting protein is the chloroplast-localized enzyme N-acetyl glutamate kinase, which catalyzes the key regulatory step in the pathway to arginine biosynthesis. The interaction of PII with N-acetyl glutamate kinase was confirmed through pull-down, gel filtration, and isothermal titration calorimetry experiments, and binding was shown to be enhanced in the presence of the downstream product, arginine. Enzyme kinetic analysis showed that PII increases N acetyl glutamate kinase activity slightly, but the primary function of binding is to relieve inhibition of enzyme activity by the pathway product, arginine. Knowing the identity of PII-binding proteins across a spectrum of photosynthetic and non-photosynthetic organisms provides a framework for a more complete understanding of the function of this highly conserved signaling protein. PMID- 16377629 TI - HER-2/neu represses the metastasis suppressor RECK via ERK and Sp transcription factors to promote cell invasion. AB - Matrix metalloproteinase (MMP) inhibitory proteins may negatively regulate MMP activity to suppress tumor metastasis. In this study, we demonstrate that the HER 2/neu oncogene inhibits the expression of the MMP inhibitor RECK to promote cell invasion. RECK was inhibited via transcriptional repression in B104-1-1 cells, which express constitutively active HER-2/neu. Overexpression of HER-2/neu in NIH/3T3 or HaCaT cells also suppressed RECK expression. Deletion and mutation assays showed that HER-2/neu repressed RECK via the Sp1-binding site localized in the -82/-71 region from the translational start site. DNA affinity precipitation and chromatin immunoprecipitation assays indicated that binding of Sp1 and Sp3 to this consensus site was increased in B104-1-1 cells. We also found that HER-2/neu inhibited RECK via the ERK signaling pathway. Sp1 proteins phosphorylated at Thr453 and Thr739 by ERK bound preferentially to the RECK promoter, and this binding was reversed by HER-2/neu and ERK inhibitors. Furthermore, our data indicate that HER-2/neu obviously increased HDAC1 binding to the Sp1-binding site localized in the -82/-71 region of the RECK promoter. The histone deacetylase inhibitor trichostatin A reversed HER-2/neu-induced inhibition of RECK. HER-2/neu activation was associated with increased MMP-9 secretion and activation. Re expression of RECK in HER-2/neu-overexpressing cells inhibited MMP-9 secretion and cell invasion. Taken together, our results suggest that HER-2/neu induces the binding of Sp proteins and HDAC1 to the RECK promoter to inhibit RECK expression and to promote cell invasion. Restoration of RECK provides a novel strategy for the inhibition of HER-2/neu-induced metastasis. PMID- 16377630 TI - Phenotypes of mice lacking extracellular superoxide dismutase and copper- and zinc-containing superoxide dismutase. AB - Mice lacking the secreted extracellular superoxide dismutase (EC-SOD) or the cytosolic copper- and zinc-containing SOD (CuZn-SOD) show relatively mild phenotypes. To explore the possibility that the isoenzymes have partly overlapping functions, single and double knockout mice were examined. The absence of EC-SOD was found to be without effect on the lifespan of mice, and the reduced lifespan of CuZn-SOD knockouts was not further shortened by EC-SOD deficiency. The urinary excretion of isoprostanes was increased in CuZn-SOD knockout mice, and plasma thiobarbituric acid-reactive substances levels were elevated in EC-SOD knockout mice. These oxidant stress markers showed potentiated increases in the absence of both isoenzymes. Other alterations were mainly found in CuZn-SOD knockout mice, such as halved glutathione peroxidase activity in the tissues examined and increased glutathione and iron in the liver. There were no changes in tissue content of the alternative superoxide scavenger ascorbate, but there was a 25% reduction in ascorbate in blood plasma in mice lacking CuZn-SOD. No increase was found in the urinary excretion of the terminal metabolites of NO, nitrite, and nitrate in any of the genotypes. In conclusion, apart from the increases in the global urinary and plasma oxidant stress markers, our phenotype studies revealed no other evidence that the copper- and zinc-containing SOD isoenzymes have overlapping roles. PMID- 16377631 TI - Characterization of a novel interaction between ELMO1 and ERM proteins. AB - ERMs are closely related proteins involved in cell migration, cell adhesion, maintenance of cell shape, and formation of microvilli through their ability to cross-link the plasma membrane with the actin cytoskeleton. ELMO proteins are also known to regulate actin cytoskeleton reorganization through activation of the small GTPbinding protein Rac via the ELMO-Dock180 complex. Here we showed that ERM proteins associate directly with ELMO1 as purified recombinant proteins in vitro and at endogenous levels in intact cells. We mapped ERM binding on ELMO1 to the N-terminal 280 amino acids, which overlaps with the region required for binding to the GTPase RhoG, but is distinct from the C-terminal Dock180 binding region. Consistent with this, ELMO1 could simultaneously bind both radixin and Dock180, although radixin did not alter Rac activation via the Dock180-ELMO complex. Most interestingly, radixin binding did not affect ELMO binding to active RhoG and a trimeric complex of active RhoG-ELMO-radixin could be detected. Moreover, the three proteins colocalized at the plasma membrane. Finally, in contrast to most other ERM-binding proteins, ELMO1 binding occurred independently of the state of radixin C-terminal phosphorylation, suggesting an ELMO1 interaction with both the active and inactive forms of ERM proteins and implying a possible role of ELMO in localizing or retaining ERM proteins in certain cellular sites. Together these data suggest that ELMO1-mediated cytoskeletal changes may be coordinated with ERM protein crosslinking activity during dynamic cellular functions. PMID- 16377632 TI - Mucin granule intraluminal organization in living mucous/goblet cells. Roles of protein post-translational modifications and secretion. AB - Recent studies suggest that the mucin granule lumen consists of a matrix meshwork embedded in a fluid phase. Secretory products can both diffuse, although very slowly, through the meshwork pores and interact noncovalently with the matrix. Using a green fluorescent protein-mucin fusion protein (SHGFP-MUC5AC/CK) as a FRAP (fluorescence recovery after photobleaching) probe, we have assessed in living mucous cells the relative importance of different protein post translational modifications on the intragranular organization. Long term inhibition of mucin-type O-glycosylation, sialylation, or sulfation altered SHGFP MUC5AC/CK characteristic diffusion time (t(1/2)), whereas all but sulfation diminished its mobile fraction. Reduction of protein disulfide bonds with tris(hydroxypropyl)phosphine resulted in virtually complete immobilization of the SHGFP-MUC5AC/CK intragranular pool. However, when activity of the vacuolar H+ ATPase was also inhibited, disulfide reduction decreased SHGFP-MUC5AC/CK t((1/2)) while diminishing its intraluminal concentration. Similar FRAP profiles were observed in granules that remained in the cells after the addition of a mucin secretagogue. Taken together these results suggest that: (a) the relative content of O-glycans and intragranular anionic groups is crucial for protein diffusion through the intragranular meshwork; (b) protein-protein, rather than carbohydrate mediated, interactions are responsible for binding of SHGFP-MUC5AC/CK to the immobile fraction, although the degree of matrix O-glycosylation and sialylation affects such interactions; (c) intragranular organization does not depend on covalent multimerization of mucins or the presence of native disulfide bonds in the intragranular mucin/proteins, but rather on specific protein-mediated interactions that are important during the early stages of mucin matrix condensation; (d) alterations of the intragranular matrix precede granule discharge, which can be partial and, accordingly, does not necessarily involve the disappearance of the granule. PMID- 16377633 TI - A molecular determinant of nickel inhibition in Cav3.2 T-type calcium channels. AB - Molecular cloning studies have revealed that heterogeneity of T-type Ca2+ currents in native tissues arises from the three isoforms of Ca(v)3 channels: Ca(v)3.1, Ca(v)3.2, and Ca(v)3.3. From pharmacological analysis of the recombinant T-type channels, low concentrations (<50 microM) of nickel were found to selectively block the Ca(v)3.2 over the other isoforms. To date, however, the structural element(s) responsible for the nickel block on the Ca(v)3.2 T-type Ca2+ channel remain unknown. Thus, we constructed chimeric channels between the nickel-sensitive Ca(v)3.2 and the nickel-insensitive Ca(v)3.1 to localize the region interacting with nickel. Systematic assaying of serial chimeras suggests that the region preceding domain I S4 of Ca(v)3.2 contributes to nickel block. Point mutations of potential nickel-interacting sites revealed that H191Q in the S3-S4 loop of domain I significantly attenuated the nickel block of Ca(v)3.2, mimicking the nickel-insensitive blocking potency of Ca(v)3.1. These findings indicate that His-191 in the S3-S4 loop is a critical residue conferring nickel block to Ca(v)3.2 and reveal a novel role for the S3-S4 loop to control ion permeation through T-type Ca2+ channels. PMID- 16377634 TI - Processivity of chimeric class V myosins. AB - Unconventional myosin V takes many 36-nm steps along an actin filament before it dissociates, thus ensuring its ability to move cargo intracellularly over long distances. In the present study we assessed the structural features that affect processive run length by analyzing the properties of chimeras of mouse myosin V and a non-processive class V myosin from yeast (Myo4p) (Reck-Peterson, S. L., Tyska, M. J., Novick, P. J., and Mooseker, M. S. (2001) J. Cell Biol. 153, 1121 1126). Surprisingly a chimera containing the yeast motor domain on the neck and rod of mouse myosin V (Y-MD) showed longer run lengths than mouse wild type at low salt. Run lengths of mouse myosin V showed little salt dependence, whereas those of Y-MD decreased steeply with ionic strength, similar to a chimera containing yeast loop 2 in the mouse myosin V backbone. Loop 2 binds to acidic patches on actin in the weak binding states of the cycle (Volkmann, N., Liu, H., Hazelwood, L., Krementsova, E. B., Lowey, S., Trybus, K. M., and Hanein, D. (2005) Mol. Cell 19, 595-605). Constructs containing yeast loop 2, which has no net charge compared with +6 for wild type, showed a higher K(m) for actin in steady-state ATPase assays. The results imply that a positively charged loop 2 and a high affinity for actin are important to maintain processivity near physiologic ionic strength. PMID- 16377635 TI - The smaller isoforms of ankyrin 3 bind to the p85 subunit of phosphatidylinositol 3'-kinase and enhance platelet-derived growth factor receptor down-regulation. AB - The Src homology 2 (SH2) domains of the p85 subunit of phosphatidylinositol 3' kinase have been shown to bind to the tyrosine-phosphorylated platelet-derived growth factor receptor (PDGFR). Previously, we have demonstrated that p85 SH2 domains can also bind to the serine/threonine kinase A-Raf via a unique phosphorylation-independent interaction. In this report, we describe a new phosphotyrosine-independent p85 SH2-binding protein, ankyrin 3 (Ank3). In general, ankyrins serve a structural role by binding to both integral membrane proteins at the plasma membrane and spectrin/fodrin proteins of the cytoskeleton. However, smaller isoforms of Ank3 lack the membrane domain and are localized to late endosomes and lysosomes. We found that p85 binds directly to these smaller 120- and 105-kDa Ank3 isoforms. Both the spectrin domain and the regulatory domain of Ank3 are involved in binding to p85. At least two domains of p85 can bind to Ank3, and the interaction involving the p85 C-SH2 domain was found to be phosphotyrosine-independent. Overexpression of the 120- or 105-kDa Ank3 proteins resulted in significantly enhanced PDGFR degradation and a reduced ability to proliferate in response to PDGF. Ank3 overexpression also differentially regulated signaling pathways downstream from the PDGFR. Chloroquine, an inhibitor of lysosomal-mediated degradation pathways, blocked the ability of Ank3 to enhance PDGFR degradation. Immunofluorescence experiments demonstrated that both small Ank3 isoforms colocalized with the lysosomal-associated membrane protein and with p85 and the PDGFR. These results suggest that Ank3 plays an important role in lysosomal-mediated receptor down-regulation, likely through a p85-Ank3 interaction. PMID- 16377636 TI - Formation of tau inclusions in knock-in mice with familial Alzheimer disease (FAD) mutation of presenilin 1 (PS1). AB - Mutations in the presenilin 1 (PS1) gene are responsible for the early onset of familial Alzheimer disease (FAD). Accumulating evidence shows that PS1 is involved in gamma-secretase activity and that FAD-associated mutations of PS1 commonly accelerate Abeta(1-42) production, which causes Alzheimer disease (AD). Recent studies suggest, however, that PS1 is involved not only in Abeta production but also in other processes that lead to neurodegeneration. To better understand the causes of neurodegeneration linked to the PS1 mutation, we analyzed the development of tau pathology, another key feature of AD, in PS1 knock-in mice. Hippocampal samples taken from FAD mutant (I213T) PS1 knock-in mice contained hyperphosphorylated tau that reacted with various phosphodependent tau antibodies and with Alz50, which recognizes the conformational change of PHF tau. Some neurons exhibited Congo red birefringence and Thioflavin T reactivity, both of which are histological criteria for neurofibrillary tangles (NFTs). Biochemical analysis of the samples revealed SDS-insoluble tau, which under electron microscopy examination, resembled tau fibrils. These results indicate that our mutant PS1 knock-in mice exhibited NFT-like tau pathology in the absence of Abeta deposition, suggesting that PS1 mutations contribute to the onset of AD not only by enhancing Abeta(1-42) production but by also accelerating the formation and accumulation of filamentous tau. PMID- 16377637 TI - Dynamics of the upper 50-kDa domain of myosin V examined with fluorescence resonance energy transfer. AB - The upper 50-kDa region of myosin may be critical for coupling between the nucleotide- and actin-binding regions. We introduced a tetracysteine motif in the upper 50-kDa domain (residues 292-297) of myosin V containing a single IQ domain (MV 1IQ), allowing us to label this site with the fluorescein biarscenical hairpin-binding dye (FlAsH) (MV 1IQ FlAsH). The enzymatic properties of MV 1IQ FlAsH were similar to those of unlabeled MV 1IQ except for a 3-fold reduced ADP release rate. MV 1IQ FlAsH was also capable of moving actin filaments in the in vitro motility assay. To examine rotation of the upper 50-kDa region, we determined the difference in the degree of energy transfer from N methylanthraniloyl (mant)-labeled nucleotides to FlAsH in both steady-state and transient kinetic experiments. The energy transfer efficiency was higher with mant-ATP (0.65 +/- 0.02) compared with mant-ADP (0.55 +/- 0.02) in the absence of actin. Stopped-flow measurements suggested that the energy transfer efficiency decreased with phosphate release (0.04 s(-1)) in the absence of actin. In contrast, upon mixing MV 1IQ FlAsH in the ADP.P(i) state with actin, a decrease in the energy transfer signal was observed at a rate of 13 s(-1), similar to the ADP release rate. Our results demonstrate there was no change in the energy transfer signal upon actin-activated phosphate release and suggest that actin binding alters the dynamics of the upper 50-kDa region, which may be critical for the ability of myosin to bind tightly to both ADP and actin. PMID- 16377638 TI - Suberoylanilide hydroxamic acid potentiates apoptosis, inhibits invasion, and abolishes osteoclastogenesis by suppressing nuclear factor-kappaB activation. AB - Because of its ability to suppress tumor cell proliferation, angiogenesis, and inflammation, the histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) is currently in clinical trials. How SAHA mediates its effects is poorly understood. We found that in several human cancer cell lines, SAHA potentiated the apoptosis induced by tumor necrosis factor (TNF) and chemotherapeutic agents and inhibited TNF-induced invasion and receptor activator of NF-kappaB ligand induced osteoclastogenesis, all of which are known to require NF-kappaB activation. These observations corresponded with the down-regulation of the expression of anti-apoptotic (IAP1, IAP2, X chromosome-linked IAP, Bcl-2, Bcl x(L), TRAF1, FLIP, and survivin), proliferative (cyclin D1, cyclooxygenase 2, and c-Myc), and angiogenic (ICAM-1, matrix metalloproteinase-9, and vascular endothelial growth factor) gene products. Because several of these genes are regulated by NF-kappaB, we postulated that SAHA mediates its effects by modulating NF-kappaB and found that SAHA suppressed NF-kappaB activation induced by TNF, IL-1beta, okadaic acid, doxorubicin, lipopolysaccharide, H(2)O(2), phorbol myristate acetate, and cigarette smoke; the suppression was not cell type specific because both inducible and constitutive NF-kappaB activation was inhibited. We also found that SAHA had no effect on direct binding of NF-kappaB to the DNA but inhibited sequentially the TNF-induced activation of IkappaBalpha kinase, IkappaBalpha phosphorylation, IkappaBalpha ubiquitination, IkappaBalpha degradation, p65 phosphorylation, and p65 nuclear translocation. Furthermore, SAHA inhibited the NF-kappaB-dependent reporter gene expression activated by TNF, TNFR1, TRADD, TRAF2, NF-kappaB-inducing kinase, IkappaBalpha kinase, and the p65 subunit of NF-kappaB. Overall, our results indicated that NF-kappaB and NF-kappaB regulated gene expression inhibited by SAHA can enhance apoptosis and inhibit invasion and osteoclastogenesis. PMID- 16377639 TI - Adipose tissue proadipogenic redox changes in obesity. AB - The role of inflammation and oxidative stress in the development of obesity and associated metabolic disorders is under debate. We investigated the redox metabolism in a non-diabetic obesity model, i.e. 11-week-old obese Zucker rats. Antioxidant enzyme activities, lipophilic antioxidant (alpha-tocopherol, coenzymes Q) and hydrophilic antioxidant (glutathione, vitamin C) contents and their redox state (% oxidized form), were studied in inguinal white fat and compared with blood and liver. The adipose tissues of obese animals showed a specific higher content of hydrophilic molecules in a lower redox state than those of lean animals, which were associated with lower lipophilic molecule content and lipid peroxidation. Conversely and as expected, glutathione content decreased and its redox state increased in adipose tissues of rats subjected to lipopolysaccharide-induced systemic oxidative stress. In these in vivo models, oxidative stress and obesity thus had opposite effects on adipose tissue redox state. Moreover, the increase in glutathione content and the decrease of its redox state by antioxidant treatment promoted in vitro the accumulation of triglycerides in preadipocytes. Taken together and contrary to the emergent view, our results suggest that obesity is associated with an intracellular reduced redox state that promotes on its own the development of a deleterious proadipogenic process. PMID- 16377640 TI - Stanniocalcin 1 acts as a paracrine regulator of growth plate chondrogenesis. AB - During embryogenesis, the expression of mammalian stanniocalcin (STC1) in the appendicular skeleton suggests its involvement in the regulation of longitudinal bone growth. Such a role is further supported by the presence of dwarfism in mice overexpressing STC1. Yet, the STC 1 inhibitory effect on growth may be related to both postnatal metabolic abnormalities and prenatal defective bone formation. In our study, we used an organ culture system to evaluate the effects of STC on growth plate chondrogenesis, which is the primary determinant of longitudinal bone growth. Fetal rat metatarsal bones were cultured in the presence of recombinant human STC (rhSTC). After 3 days, rhSTC suppressed metatarsal growth, growth plate chondrocyte proliferation and hypertrophy/differentiation, and extracellular matrix synthesis. In addition, rhSTC increased the number of apoptotic chondrocytes in the growth plate. In cultured chondrocytes, rhSTC increased phosphate uptake, reduced chondrocyte proliferation and matrix synthesis, and induced apoptosis. All these effects were reversed by culturing chondrocytes with rhSTC and phosphonoformic acid, an inhibitor of phosphate transport. The rhSTC-mediated inhibition of metatarsal growth and growth plate chondrocyte proliferation and hypertrophy/differentiation was abolished by culturing metatarsals with rhSTC and phosphonoformic acid. Taken together, our findings indicate that STC1 inhibits longitudinal bone growth directly at the growth plate. Such growth inhibition, likely mediated by an increased chondrocyte phosphate uptake, results from suppressed chondrocyte proliferation, hypertrophy/differentiation, and matrix synthesis and by increased apoptosis. Last, the expression of both STC1 and its binding site in the growth plate would support an autocrine/paracrine role for this growth factor in the regulation of growth plate chondrogenesis. PMID- 16377641 TI - The crystal structure of (S)-3-O-geranylgeranylglyceryl phosphate synthase reveals an ancient fold for an ancient enzyme. AB - We report crystal structures of the citrate and sn-glycerol-1-phosphate (G1P) complexes of (S)-3-O-geranylgeranylglyceryl phosphate synthase from Archaeoglobus fulgidus (AfGGGPS) at 1.55 and 2.0 A resolution, respectively. AfGGGPS is an enzyme that performs the committed step in archaeal lipid biosynthesis, and it presents the first triose phosphate isomerase (TIM)-barrel structure with a prenyltransferase function. Our studies provide insight into the catalytic mechanism of AfGGGPS and demonstrate how it selects for the sn-G1P isomer. The replacement of "Helix 3" by a "strand" in AfGGGPS, a novel modification to the canonical TIM-barrel fold, suggests a model of enzyme adaptation that involves a "greasy slide" and a "swinging door." We propose functions for the homologous PcrB proteins, which are conserved in a subset of pathogenic bacteria, as either prenyltransferases or being involved in lipoteichoic acid biosynthesis. Sequence and structural comparisons lead us to postulate an early evolutionary history for AfGGGPS, which may highlight its role in the emergence of Archaea. PMID- 16377642 TI - Single amino acid mutations alter the distribution of human porphobilinogen synthase quaternary structure isoforms (morpheeins). AB - Porphobilinogen synthase (PBGS) is an obligate oligomer that can exist in functionally distinct quaternary states of different stoichiometries, which are called morpheeins. The morpheein concept describes an ensemble of quaternary structure isoforms wherein different structures of the monomer dictate different multiplicities of the oligomer (Jaffe, E. K. (2005) Trends Biochem. Sci. 30, 490 497). Human PBGS assembles into long-lived morpheeins and has been shown to be capable of forming either a high activity octamer or a low activity hexamer (Breinig, S., Kervinen, J., Stith, L., Wasson, A. S., Fairman, R., Wlodawer, A., Zdanov, A., and Jaffe, E. K. (2003) Nat. Struct. Biol. 10, 757-763). All PBGS monomers contain an alphabeta-barrel domain and an N-terminal arm domain. The N terminal arm structure varies among PBGS morpheeins, and the spatial relationship between the arm and the barrel dictates the different quaternary assemblies. We have analyzed the structures of human PBGS morpheeins for key interactions that would be predicted to affect the oligomeric assembly. Examples of individual mutations that shift assembly of human PBGS away from the native octamer are R240A and W19A. The alternate morpheeins of human PBGS variants R240A and W19A are chromatographically separable from each other and kinetically distinct; their structure and dynamics have been characterized by native gel electrophoresis, dynamic light scattering, and analytical ultracentrifugation. R240A assembles into a metastable hexamer, which can undergo a reversible conversion to the octamer in the presence of substrate. The metastable nature of the R240A hexamer supports the hypothesis that octameric and hexameric morpheeins of PBGS are very close in energy. W19A assembles into a mixture of dimers, which appear to be stable. PMID- 16377643 TI - Didgeridoo playing as alternative treatment for obstructive sleep apnoea syndrome: randomised controlled trial. AB - OBJECTIVE: To assess the effects of didgeridoo playing on daytime sleepiness and other outcomes related to sleep by reducing collapsibility of the upper airways in patients with moderate obstructive sleep apnoea syndrome and snoring. DESIGN: Randomised controlled trial. SETTING: Private practice of a didgeridoo instructor and a single centre for sleep medicine. PARTICIPANTS: 25 patients aged > 18 years with an apnoea-hypopnoea index between 15 and 30 and who complained about snoring. INTERVENTIONS: Didgeridoo lessons and daily practice at home with standardised instruments for four months. Participants in the control group remained on the waiting list for lessons. MAIN OUTCOME MEASURE: Daytime sleepiness (Epworth scale from 0 (no daytime sleepiness) to 24), sleep quality (Pittsburgh quality of sleep index from 0 (excellent sleep quality) to 21), partner rating of sleep disturbance (visual analogue scale from 0 (not disturbed) to 10), apnoea-hypopnoea index, and health related quality of life (SF-36). RESULTS: Participants in the didgeridoo group practised an average of 5.9 days a week (SD 0.86) for 25.3 minutes (SD 3.4). Compared with the control group in the didgeridoo group daytime sleepiness (difference -3.0, 95% confidence interval 5.7 to -0.3, P = 0.03) and apnoea-hypopnoea index (difference -6.2, -12.3 to 0.1, P = 0.05) improved significantly and partners reported less sleep disturbance (difference -2.8, -4.7 to -0.9, P < 0.01). There was no effect on the quality of sleep (difference -0.7, -2.1 to 0.6, P = 0.27). The combined analysis of sleep related outcomes showed a moderate to large effect of didgeridoo playing (difference between summary z scores -0.78 SD units, -1.27 to -0.28, P < 0.01). Changes in health related quality of life did not differ between groups. CONCLUSION: Regular didgeridoo playing is an effective treatment alternative well accepted by patients with moderate obstructive sleep apnoea syndrome. Trial registration ISRCTN: 31571714. PMID- 16377644 TI - Detection of secretory IgA antibodies against gliadin and human tissue transglutaminase in stool to screen for coeliac disease in children: validation study. AB - OBJECTIVE: To evaluate two commercial stool tests for detection of secretory IgA antibodies against gliadin and human tissue transglutaminase for diagnosis of coeliac disease in children with symptoms. SETTING: Tertiary care children's hospital. PARTICIPANTS: Coded stool samples from 20 children with newly diagnosed coeliac disease and 64 controls. Six children with coeliac disease had stool tests every two weeks for three months after starting a gluten-free diet. MAIN OUTCOME MEASURES: Secretory IgA antibodies against gliadin and human tissue transglutaminase in stool samples, determined in duplicate by using recommended cut-off limits. RESULTS: Sensitivity of faecal antibodies against human tissue transglutaminase was 10% (95% confidence interval 1% to 32%), and specificity was 98% (91% to 100%). For antibodies against gliadin, sensitivity was 6% (0% to 29%) and specificity was 97% (89% to 100%). Optimisation of cut-off limits by receiver operating characteristic analysis and use of results of both tests increased sensitivity to 82%, but specificity decreased to 58%. All follow-up stool tests remained negative, except for two positive anti-gliadin results in one patient, six and 10 weeks after the gluten-free diet was started. CONCLUSIONS: Neither stool test was suitable for screening for coeliac disease in children with symptoms. PMID- 16377645 TI - The effects of continuous epidural analgesia on Doppler velocimetry of uterine arteries during different periods of labour analgesia. AB - BACKGROUND: The transient effects of epidural bupivacaine 0.25-0.5% on the Doppler velocimetry of umbilical and uterine arteries had been reported, but the effects of continuous lower dose epidural bupivacaine (0.05-0.1%) infusion for labour analgesia have never been reported. In this study, we evaluated the effects of continuous epidural bupivacaine 0.075% on the Doppler velocimetry of uterine arteries. METHODS: Twenty pregnant women for labour analgesia received continuous epidural bupivacaine 0.075% infusion. We used a 4-MHz continuous-wave Doppler probe (Multigon 500A) with a 200 Hz thump filter to detect uterine blood flow velocity. We recorded the velocimetry data for uterine relaxation and contraction during five time periods: pre-epidural insertion, 1, 2, and 4 h post epidural infusion, and after delivery of fetus. RESULTS: Our data showed that the velocimetric indices of uterine vascular resistance were significantly increased 1, 2, and 4 h after epidural infusion when compared with the pre-epidural level; these returned to the baseline after delivery. CONCLUSION: Continuous epidural analgesia with bupivacaine 0.075% increases the resistance of uterine artery and therefore possibly reduces the uterine blood flow. PMID- 16377646 TI - Chemical encapsulation of rocuronium by synthetic cyclodextrin derivatives: reversal of neuromuscular block in anaesthetized Rhesus monkeys. AB - BACKGROUND: At present, reversal of neuromuscular block induced by steroidal neuromuscular blocking agents (NMBAs) is achieved by administration of cholinesterase inhibitors. Chemical encapsulation of steroidal NMBAs, such as rocuronium, by a cyclodextrin is a new concept in neuromuscular block reversal. The present study evaluates the capacity of nine synthetic cyclodextrin derivatives (Org 25288, Org 25289, Org 25467, Org 25168, Org 25169, Org 25555, Org 25166, Org 26142, and Org 25969) to reverse constant neuromuscular block of approximately 90%, induced by rocuronium infusion in the Rhesus monkey, using single twitch stimulation. The ability of these cyclodextrin derivatives to reverse neuromuscular block was compared with the reversal of the same neuromuscular block by the commonly used combination of neostigmine and atropine. METHODS: After a bolus injection of rocuronium, continuous infusion was started to reduce twitch contractions to approximately 10% of baseline values. After a steady state block of at least 10 min the infusion was stopped and the preparation was allowed to recover spontaneously. This process was repeated, but at the time the infusion was stopped, either one of the nine cyclodextrin derivatives or a combination of neostigmine and atropine was given. RESULTS: Recovery with cyclodextrin derivatives Org 26142 and Org 25969 was faster than after a combination of neostigmine and atropine (P<0.05). Injection of these cyclodextrin derivatives did not affect blood pressure or heart rate. Signs of residual block or recurarization were not observed in any of these experiments. In the experiments in which a combination of neostigmine and atropine was given, two animals showed signs of abdominal discomfort as frequently seen after the administration of neostigmine and significant changes in circulatory variables. CONCLUSIONS: Chemical encapsulation or chelation of rocuronium is a new concept in reversing neuromuscular block induced by rocuronium. PMID- 16377647 TI - Ephedrine and phenylephrine for the treatment of maternal hypotension in a chronic sheep model of increased placental vascular resistance. AB - BACKGROUND: We hypothesized that ephedrine and phenylephrine are equal with respect to uterine and placental haemodynamics and fetal acid-base status after exposure to maternal hypoxaemia and hypotension in a chronic sheep model of increased placental vascular resistance (R(UA)). METHODS: At 114-135 days gestation, chronically instrumented fetal sheep underwent placental embolization leading to increased R(UA). Twenty-four hours after embolization, the ewes were anaesthetized and randomized to receive boluses of ephedrine (n=7) or phenylephrine (n=6) for epidural-induced hypotension after maternal hypoxaemia. Uterine (Q(UtA)) and placental (Q(UA)) volume blood flows and uterine vascular resistance (R(UtA)) and R(UA) were recorded. Uterine (PI(UtA)) and umbilical artery (PI(UA)) pulsatility indices were obtained by Doppler ultrasonography. Fetal arterial blood samples were analysed for acid-base values and lactate concentrations. RESULTS: During hypotension, Q(UtA), fetal pH, BE, and Po(2) decreased whereas R(UtA), PI(UtA), R(UA), and fetal lactate concentration increased. With ephedrine, Q(UtA), R(UtA), PI(UtA), R(UA), and fetal Po(2) returned to baseline. Fetal pH, BE, and lactate concentration did not change from hypotensive values. With phenylephrine, Q(UtA) remained lower (P=0.007) and R(UtA) (P=0.007), PI(UtA) (P=0.013), and R(UA) (P=0.050) higher than at baseline. Fetal Po(2) returned to baseline and fetal pH and BE did not change from hypotensive values. However, fetal lactate concentration increased further (mean difference 1.49, 95% confidence interval 0.72-2.26 mmol litre(-1); P=0.004). CONCLUSIONS: In a chronic sheep model of increased placental vascular resistance, compared with ephedrine administration, phenylephrine administration was associated with impaired uterine and placental haemodynamics and increased fetal lactate concentrations. PMID- 16377648 TI - Use of percutaneous cardiopulmonary support in catastrophic massive pulmonary fat embolism. AB - We report a patient who sustained massive catastrophic pulmonary fat embolism while undergoing surgery for intramedullary nail of femur. Application of percutaneous cardiopulmonary support, in our view, saved the patient from cardiopulmonary collapse. PMID- 16377649 TI - Bedside red cell volumetry by low-dose carboxyhaemoglobin dilution using expiratory gas analysis. AB - BACKGROUND: We developed a non-invasive, continuous, high-resolution method of measuring carboxyhaemoglobin fraction (COHb%) using expiratory gas analysis (EGA). We assessed whether application of EGA to carboxyhaemoglobin dilution provides red cell volume (RCV) measurement with accuracy equivalent to that of CO haemoximetry, with a smaller infusion volume of carbon-monoxide-saturated autologous blood (COB). Method. We assessed the agreement between repeated COHb% measurements by EGA and simultaneous measurement by CO-haemoximetry, using Bland and Altman plot, in healthy subjects and patients with artificially controlled ventilation and no radiological evidence of pulmonary oedema or atelectasis. We assessed the agreement between RCV measurements by EGA with infusion of 20 ml of COB (RCVEGA) and RCV measurements by CO-haemoximetry with infusion of 100 ml of COB (RCVHEM), in healthy subjects. RESULTS: The 'limits of agreement' between COHb% measurement by EGA (1 min average) and CO-haemoximetry were -0.09 and 0.08% in healthy subjects, and -0.11 and 0.09% in patients. Given the resolution of CO haemoximetry (0.1%), the accuracy of EGA was equivalent to or greater than that of CO-haemoximetry. The 'limits of agreement' between RCVEGA and RCVHEM were 0.14 and 0.15 litre. Given the average resolution of RCVHEM (0.14 litre), the accuracy of RCVEGA was equivalent to that of RCVHEM. CONCLUSION: EGA provided non invasive, accurate, continuous, high-resolution COHb% measurements. Applying EGA to carboxyhaemoglobin dilution, we achieved RCV measurements with accuracy equivalent to that of CO-haemoximetry, with one-fifth of the COB infusion volume. However, clinical application of the method is limited to patients with no radiological evidence of pulmonary oedema or atelectasis. PMID- 16377650 TI - Anaesthesia for brachytherapy--51/2 yr of experience in 1622 procedures. AB - BACKGROUND: Brachytherapy presents the anaesthetist with unique problems. Information on anaesthesia for brachytherapy, however, is limited. The aim of this paper is to report on our experience involving a large number of brachytherapy procedures. METHODS: A retrospective analysis of records of 1622 anaesthetic procedures in 952 patients is presented. Records were analysed in respect of patient data, tumour localization, brachytherapy treatment and the type and duration of anaesthetic procedures. RESULTS: More than one-third of patients were at high risk (ASA III or IV) and 40% were more than 60 yr. Repetitive treatments were performed on half of the patients. Breast cancer was the most common indication. The average duration of anaesthesia for pelvic brachytherapy was more than 3 h, with a high degree of variability. Regional anaesthesia was used in 30% of all cases and was the predominant technique for pelvic brachytherapy. Spinal catheter techniques represented a high proportion of those receiving regional anaesthesia. Complications resulting from regional and general anaesthesia were minor and no serious incidents occurred. CONCLUSIONS: Based on a large number of procedures, this study gives an example of anaesthetic management in brachytherapy. A substantial minority of patients would be considered high risk for surgical intervention. Regional anaesthesia was the principal technique used when dealing with tumours of the lower body. PMID- 16377651 TI - Nitrous oxide depresses electroencephalographic responses to repetitive noxious stimulation in the rat. AB - BACKGROUND: Although N(2)O has been widely used as an anaesthetic adjuvant its effect on electroencephalographic (EEG) activity is poorly understood because it is usually studied in the presence of additional anaesthetics, including inhaled anaesthetics. We examined the EEG effects of N(2)O in rats using a hyperbaric chamber that permitted N(2)O to be the sole anaesthetic. METHODS: Rats (n=10) were anaesthetized with isoflurane and EEG activity was recorded from skull screws. The rats were placed into a hyperbaric chamber and mechanically ventilated. Isoflurane was eliminated while the chamber was pressurized with N(2)O. The minimum alveolar concentration (MAC) was determined in five rats by adjusting the chamber pressure and N(2)O concentration, and applying a tetanic noxious stimulus to the tail via an electrical pass-through. EEG responses to noxious stimulation (20 electrical pulses at 40 V applied to the tail at 0.1, 1 and 3 Hz, and 50 Hz tetanic stimulation at 60 mA applied for 30 s) were determined at 1.5 and 2 atm N(2)O. RESULTS: The N(2)O MAC was 1.7+/-0.1 atm. No consistent EEG activation occurred during electrical stimulation at either partial pressure of N(2)O, although spontaneous EEG activation often occurred. Blood pressure increased after the 3 and 50 Hz stimuli. Four other rats anaesthetized with isoflurane had EEG activation with the 3 and 50 Hz stimuli. CONCLUSIONS: These data indicate that N(2)O at peri-MAC partial pressures prevents EEG activation resulting from noxious electrical stimulation. Unlike the situation with isoflurane, stimulus-evoked EEG activation did not occur at peri MAC anaesthetic concentrations, suggesting that N(2)O potently blocked ascending nociceptive transmission. PMID- 16377652 TI - Hypotensive epidural anaesthesia in patients with preoperative renal dysfunction undergoing total hip replacement. AB - BACKGROUND: Hypotensive anaesthesia does not impair renal function after surgery in normal patients but there are no reports of hypotensive anaesthesia in patients with chronic renal dysfunction (CRD). METHODS: From a database of 1893 consecutive patients undergoing total hip replacement (THR) under hypotensive epidural anaesthesia (HEA) from 1999 to 2004, 54 patients were identified with CRD (preoperative serum creatinine > or =124 micromol litre(-1)). Fifty matched pairs were identified for patients with normal renal function who have hypertension (n=50) or no hypertension (n=50). Changes in serum creatinine and blood urea nitrogen (BUN) were recorded daily for 3 days. Acute renal failure was defined as an increase in serum creatinine of 44 micromol litre(-1). RESULTS: The mean duration of hypotension (MAP<55 mm Hg) was 94 min (range 35-305 min). The mean age was 71 yr. All patients with a creatinine level of 124 micromol litre( 1) had a creatinine clearance of <40 ml min(-1) 1.73 m(-2) (range: 13-56). Patients with CRD received more crystalloid during surgery (1755 ml) than the other two groups (1435 ml) (P<0.001). Otherwise, all three groups were similar. No patients developed evidence of acute renal dysfunction immediately after or by 24 h after surgery. Three patients with CRD had an increase in creatinine of >44 micromol litre(-1) at 48 and 72 h after surgery in the setting of volume depletion (acute blood loss in two patients and early ileus in one). Renal function subsequently improved. CONCLUSION: HEA, per se, when carefully managed does not appear to predispose patients with CRD to acute renal failure after THR. PMID- 16377653 TI - Radiation of pollination systems in the Iridaceae of sub-Saharan Africa. AB - BACKGROUND: Seventeen distinct pollination systems are known for genera of sub Saharan African Iridaceae and recurrent shifts in pollination system have evolved in those with ten or more species. Pollination by long-tongued anthophorine bees foraging for nectar and coincidentally acquiring pollen on some part of their bodies is the inferred ancestral pollination strategy for most genera of the large subfamilies Iridoideae and Crocoideae and may be ancestral for the latter. Derived strategies include pollination by long-proboscid flies, large butterflies, night-flying hovering and settling moths, hopliine beetles and sunbirds. Bee pollination is diverse, with active pollen collection by female bees occurring in several genera, vibratile systems in a few and non-volatile oil as a reward in one species. Long-proboscid fly pollination, which is apparently restricted to southern Africa, includes four separate syndromes using different sets of flies and plant species in different parts of the subcontinent. Small numbers of species use bibionid flies, short-proboscid flies or wasps for their pollination; only about 2 % of species use multiple pollinators and can be described as generalists. SCOPE: Using pollination observations for 375 species and based on repeated patterns of floral attractants and rewards, we infer pollination mechanisms for an additional 610 species. Matching pollination system to phylogeny or what is known about species relationships based on shared derived features, we infer repeated shifts in pollination system in some genera, as frequently as one shift for every five or six species of southern African Babiana or Gladiolus. Specialized systems using pollinators of one pollination group, or even a single pollinator species are the rule in the family. Shifts in pollination system are more frequent in genera of Crocoideae that have bilaterally symmetric flowers and a perianth tube, features that promote adaptive radiation by facilitating precise shifts in pollen placement, in conjunction with changes in flower colour, scent and tube length. CONCLUSIONS: Diversity of pollination systems explains in part the huge species diversity of Iridaceae in sub-Saharan Africa, and permits species packing locally. Pollination shifts are, however, seen as playing a secondary role in speciation by promoting reproductive isolation in peripheral, ecologically distinct populations in areas of diverse topography, climate and soils. Pollination of Iridaceae in Eurasia and the New World, where the family is also well represented, is poorly studied but appears less diverse, although pollination by both pollen- and oil-collecting bees is frequent and bird pollination rare. PMID- 16377654 TI - Plant sexual systems and a review of the breeding system studies in the Caatinga, a Brazilian tropical dry forest. AB - BACKGROUNDS AND AIMS: The reproductive biology of a community can provide answers to questions related to the maintenance of the intraspecific pollen flow and reproductive success of populations, sharing and competition for pollinators and also questions on conservation of natural habitats affected by fragmentation processes. This work presents, for the first time, data on the occurrence and frequency of plant sexual systems for Caatinga communities, and a review of the breeding system studies of Caatinga species. METHODS: The sexual systems of 147 species from 34 families and 91 genera occurring in three Caatinga areas in north eastern Brazil were analysed and compared with worldwide studies focusing on reproductive biology of different tropical communities. KEY RESULTS: The frequency of hermaphrodite species was 83.0 % (122 species), seven of these (or 4.8 % of the total) being heterostylous. Monoecy occurred in 9.5 % (14) of the species, and andromonoecy in 4.8 % (seven). Only 2.7 % (four) of the species were dioecious. A high percentage of hermaphrodite species was expected and has been reported for other tropical ecosystems. With respect to the breeding system studies with species of the Caatinga, the authors' data for 21 species and an additional 18 species studied by others (n = 39) revealed a high percentage (61.5 %) of obligatory self-incompatibility. Agamospermy was not recorded among the Caatinga studied species. CONCLUSIONS: The plant sexual systems in the Caatinga, despite the semi-arid climate, are similar to other tropical dry and wet forest communities, including those with high rainfall levels, except for the much lower percentage of dioecious species. The high frequency of self-incompatible species is similar to that reported for Savanna areas in Brazil, and also for dry (deciduous and semideciduous) and humid tropical forest communities. PMID- 16377656 TI - Cohort profile: the Stockholm birth cohort of 1953. PMID- 16377657 TI - Is the association between low job control and coronary heart disease confounded by risk factors measured in childhood and adolescence among Swedish males 40-53 years of age? AB - OBJECTIVES: Low job control is associated with an increased relative risk of coronary heart disease (CHD). The increased risk seems to be most marked in men under 55 years of age. Adverse social circumstances in childhood have been related to an increased risk of CHD in adulthood. The aim of this study was to investigate the role of differences in CHD risk factors measured in childhood or late adolescence (indicators of social circumstances and negative behaviour), as well as social circumstances in adulthood, for the association between low job control and CHD among Swedish men 40-53 years of age. METHODS: The study is based on a cohort of 49 323 young Swedish males, born 1949-51. For the study we have used information on childhood environment collected at the 1960 census (when the subjects were 9-11 years of age), negative behavioural factors collected at compulsory conscription for military training among young Swedish men in 1969/70 (when the subjects were 18-20 years of age), data on job control from 1990 (measured indirectly from occupational titles using a job exposure matrix, when the subjects were 39-41 years of age), and follow-up data on CHD hospitalization and mortality between the years 1991 and 2003. RESULTS: An increased relative risk of CHD (HR = 1.55; 95% CI 1.31-1.84) was found among workers with low job control, compared with workers with high job control. Risk factors for CHD measured in different phases during the life course were strongly associated with level of job control in middle age. In multivariate analyses, taking the factors measured in childhood into consideration and also adjusting for lifestyle factors measured at age 18-20 (smoking, heavy alcohol consumption, and overweight) the increased relative risk of CHD in low control jobs was reduced by 85%. After also adjusting for adult income (according to the 1985 census) no increased relative risks remained. CONCLUSION: Risk factors for CHD measured already in childhood and adolescence could explain a substantial part of the of the increased risk of CHD and mortality among males 40-53 years of age associated with the measure of low work control used in this study. The results suggest that low job control, measured as in this study, is not a risk factor for CHD among men in this age group. PMID- 16377658 TI - Cohort profile: the Metropolit 1953 Danish male birth cohort. PMID- 16377659 TI - Beta-mannosidosis mice: a model for the human lysosomal storage disease. AB - Beta-mannosidase, a lysosomal enzyme which acts exclusively at the last step of oligosaccharide catabolism in glycoprotein degradation, functions to cleave the unique beta-linked mannose sugar found in all N-linked oligosaccharides of glycoproteins. Deficiency of this enzyme results in beta-mannosidosis, a lysosomal storage disease characterized by the cellular accumulation of small oligosaccharides. In human beta-mannosidosis, the clinical presentation is variable and can be mild, even when caused by functionally null mutations. In contrast, two existing ruminant animal models have disease that is consistent and severe. To further explore the molecular pathology of this disease and to investigate potential treatment strategies, we produced a beta-mannosidase knockout mouse. Homozygous mutant mice have undetectable beta-mannosidase activity. General appearance and growth of the knockout mice are similar to the wild-type littermates. At >1 year of age, these mice exhibit no dysmorphology or overt neurological problems. The mutant animals have consistent cytoplasmic vacuolation in the central nervous system and minimal vacuolation in most visceral organs. Thin-layer chromatography demonstrated an accumulation of disaccharide in epididymis and brain. This mouse model closely resembles human beta-mannosidosis and provides a useful tool for studying the phenotypic variation in different species and will facilitate the study of potential therapies for lysosomal storage diseases. PMID- 16377660 TI - Stem-cell protein Piwil2 is widely expressed in tumors and inhibits apoptosis through activation of Stat3/Bcl-XL pathway. AB - The genes of the piwi family are defined by conserved PAZ and Piwi domains and play important roles in stem-cell self-renewal, RNA silencing and translational regulation in various organisms. Both, mouse and human Piwil2 genes, members of the piwi gene family, are specifically expressed in testis. We report here enhanced expression of the human Piwil2 gene in testicular seminomas, but not in testicular non-seminomatous tumors. Expression of the Piwil2 gene was also found in different tumors examined, including prostate, breast, gastrointestinal, ovarian and endometrial cancer of human and in breast tumors, rhabdomyosarcoma and medulloblastoma of mouse. Therefore, Piwil2 can be categorized as a novel member of cancer/testis antigens. To identify genes activated by Piwil2, RNA isolated from NIH-3T3 cells expressing constitutively Piwil2 were compared with RNA samples from control NIH-3T3 cells using a cancer gene array. Induction of high-level expression of the antiapoptotic gene Bcl-X(L) was observed in cells expressing Piwil2. Furthermore, increased Bcl-X(L) expression correlated with increase of signal transducer and activator of transcription 3 (Stat3) expression. Gene silencing of Piwil2 with its small interference RNA suppressed Stat3 and Bcl-X(L) expression and induced apoptosis. A causal link between Piwil2 expression and inhibition of apoptosis and enhanced proliferation was demonstrated in cells expressing Piwil2. Furthermore, results of soft agar assay indicated that Piwil2 overexpression induced transformation of fibroblast cells. In summary, our results demonstrate that Piwil2 is widely expressed in tumors and acts as an oncogene by inhibition of apoptosis and promotion of proliferation via Stat3/Bcl-X(L) signaling pathway. Expression of Piwil2 in a wide variety of tumors could be a useful prognostic factor that could have also diagnostic and therapeutic implications. PMID- 16377662 TI - Quinolones for treatment of human brucellosis: critical review of the evidence from microbiological and clinical studies. PMID- 16377663 TI - Microdialysis study of imipenem distribution in the intraperitoneal fluid of rats with or without experimental peritonitis. AB - The purpose of this study was to extend the use of microdialysis to the investigation of antibiotic distribution into the intraperitoneal fluid of rats with or without peritonitis. Microdialysis probes were inserted into the jugular vein and peritoneal cavity of control rats or rats with intra-abdominal sepsis (n = 8 in each group) induced by cecal ligation and punctures. Imipenem (IPM) probe recoveries were determined in each rat by retrodialysis by drug. IPM was infused intravenously at a dose of 30 mg . kg(-1) over 30 min, microdialysis samples were collected for 120 min, and IPM concentrations were determined by high-performance liquid chromatography. Intraperitoneal infection had no statistically significant effect on IPM clearance (11.9 +/- 2.3 ml.min(-1).kg(-1) in control rats versus 10.9 +/- 2.1 ml.min(-1).kg(-1) in rats with peritonitis) or the volume of distribution (296 +/- 47 ml.kg(-1) in control rats versus 310 +/- 49 ml.kg(-1) in rats with peritonitis). IPM concentration profiles in intraperitoneal fluid and blood were virtually superimposed in control rats, whereas in infected animals, the mean intraperitoneal IPM concentrations were apparently slightly lower than corresponding blood levels. However, the areas under the concentration-versus time curve estimated in intraperitoneal fluid and blood were not significantly different in both groups, with the corresponding ratios close to unity (1.01 +/- 0.19 and 0.89 +/- 0.28 in control rats and rats with peritonitis, respectively). In conclusion, IPM distribution in intraperitoneal fluid is rapid and complete both in control rats and in rats with peritonitis. PMID- 16377664 TI - Ciprofloxacin-resistant Salmonella enterica serovar Typhimurium strains are difficult to select in the absence of AcrB and TolC. AB - It has been proposed that lack of a functional efflux system(s) will lead to a lower frequency of selection of resistance to fluoroquinolones and other antibiotics. We constructed five strains of Salmonella enterica serovar Typhimurium SL1344 that lacked efflux gene components of resistance nodulation cell division pumps (acrB, acrD, acrF, acrBacrF, and tolC) plus three strains that lack genes that effect efflux gene expression (marA, soxS, and ramA) and a hypermutable strain (mutS::aph). Strains were exposed to ciprofloxacin at 2x the MIC in agar, in the presence and absence of Phe-Arg-beta-naphthylamide, an efflux pump inhibitor. Mutants were selected from all strains except those lacking acrB, tolC, or acrBacrF. For strains from which mutants were selected, there were no significant differences between the frequencies of resistance. Except for mutants of the ramA::aph strain, two phenotypes arose: resistance to quinolones only and multiple antibiotic resistance (MAR). ramA::aph mutants were resistant to quinolones only, suggesting a role for ramA in MAR in S. enterica serovar Typhimurium. Phe-Arg-beta-naphthylamide (20 microg/ml) had no effect on the frequencies of resistance or ciprofloxacin MICs. In conclusion, functional AcrB and TolC in S. enterica serovar Typhimurium are important for the selection of ciprofloxacin-resistant mutants. PMID- 16377665 TI - Multidrug-resistant Pseudomonas aeruginosa: risk factors and clinical impact. AB - Pseudomonas aeruginosa, a leading nosocomial pathogen, may become multidrug resistant (MDR). Its rate of occurrence, the individual risk factors among affected patients, and the clinical impact of infection are undetermined. We conducted an epidemiologic evaluation and molecular typing using pulsed-field gel electrophoresis (PFGE) of 36 isolates for 82 patients with MDR P. aeruginosa and 82 controls matched by ward, length of hospital stay, and calendar time. A matched case-control study identified individual risk factors for having MDR P. aeruginosa, and a retrospective matched-cohort study examined clinical outcomes of such infections. The 36 isolates belonged to 12 PFGE clones. Two clones dominated, with one originating in an intensive care unit (ICU). Cases and controls had similar demographic characteristics and numbers of comorbid conditions. A multivariate model identified ICU stay, being bedridden, having high invasive devices scores, and being treated with broad-spectrum cephalosporins and with aminoglycosides as significant risk factors for isolating MDR P. aeruginosa. Having a malignant disease was a protective factor (odds ratio [OR] = 0.2; P = 0.03). MDR P. aeruginosa was associated with severe outcomes compared to controls, including increased mortality (OR = 4.4; P = 0.04), hospital stay (hazard ratio, 2; P = 0.001), and requirement for procedures (OR = 5.4; P = 0.001). The survivors functioned more poorly at discharge than the controls, and more of the survivors were discharged to rehabilitation centers or chronic care facilities. The epidemiology of MDR P. aeruginosa is complex. Critically ill patients that require intensive care and are treated with multiple antibiotic agents are at high risk. MDR P. aeruginosa infections are associated with severe adverse clinical outcomes. PMID- 16377666 TI - High prevalence of antimicrobial resistance among Shigella isolates in the United States tested by the National Antimicrobial Resistance Monitoring System from 1999 to 2002. AB - Shigella spp. infect approximately 450,000 persons annually in the United States, resulting in over 6,000 hospitalizations. Since 1999, the National Antimicrobial Resistance Monitoring System (NARMS) for Enteric Bacteria has tested every 10th Shigella isolate from 16 state or local public health laboratories for susceptibility to 15 antimicrobial agents. From 1999 to 2002, NARMS tested 1,604 isolates. Among 1,598 isolates identified to species level, 1,278 (80%) were Shigella sonnei, 295 (18%) were Shigella flexneri, 18 (1%) were Shigella boydii, and 7 (0.4%) were Shigella dysenteriae. Overall, 1,251 (78%) were resistant to ampicillin and 744 (46%) were resistant to trimethoprim-sulfamethoxazole (TMP SMX). Prevalence of TMP-SMX- or ampicillin- and TMP-SMX-resistant Shigella sonnei isolates varied by geographic region, with lower rates in the South and Midwest regions (TMP-SMX resistance, 27% and 30%, respectively; ampicillin and TMP-SMX resistance, 25% and 22%, respectively) and higher rates in the East and West regions (TMP-SMX resistance, 66% and 80%, respectively; ampicillin and TMP-SMX resistance, 54% and 65%, respectively). Nineteen isolates (1%) were resistant to nalidixic acid (1% of S. sonnei and 2% of S. flexneri isolates); 12 (63%) of these isolates had decreased susceptibility to ciprofloxacin. One S. flexneri isolate was resistant to ciprofloxacin. All isolates were susceptible to ceftriaxone. Since 1986, resistance to ampicillin and TMP-SMX has dramatically increased. Shigella isolates in the United States remain susceptible to ciprofloxacin and ceftriaxone. PMID- 16377667 TI - Multiple combination bactericidal testing of staphylococcal biofilms from implant associated infections. AB - Standardized susceptibility testing fails to predict in vivo resistance of device related infections to antimicrobials. We assessed agents and combinations of antimicrobials against clinical isolates of Staphylococcus epidermidis and S. aureus (methicillin-resistant S. aureus and methicillin-sensitive S. aureus) retrieved from device-associated infections. Isolates were grown planktonically and as biofilms. Biofilm cultures of the organisms were found to be much more resistant to inhibitory and bactericidal effects of single and combination antibiotics than planktonic cultures (P < 0.001). Rifampin was the most common constituent of antibiotic combinations active against staphylococcal biofilms. Other frequently effective antimicrobials were vancomycin and fusidic acid. Susceptibility testing involving biofilm-associated bacteria suggests new options for combination antibiotic therapy. PMID- 16377668 TI - Comparison of censored regression and standard regression analyses for modeling relationships between antimicrobial susceptibility and patient- and institution specific variables. AB - In order to identify patients likely to be infected with resistant bacterial pathogens, analytic methods such as standard regression (SR) may be applied to surveillance data to determine patient- and institution-specific factors predictive of an increased MIC. However, the censored nature of MIC data (e.g., MIC < or = 0.5 mg/liter or MIC > 8 mg/liter) imposes certain limitations on the use of SR. In order to investigate the nature of these limitations, simulations were performed to compare a regression tailored for censored data (censored regression [CR]) and one tailored for an SR. By using a model relating piperacillin-tazobactam MICs against Enterobacter spp. to patient age and hospital bed capacity, 200 simulations of 500 isolates were performed. Various MIC censoring patterns were imposed by using 26 left- or right-censored (L,R) pairs (i.e., MICs < or = 2 mg/liter(L) [2L] or MICs > 2 mg/liter(R) [2R], respectively). Data were fit by CR and SR for which censored MICs were either (i) excluded, (ii) replaced by 2L or 2R, or (iii) replaced by 2(L - 1) or 2(R + 1). Total censoring for the 26 pairs ranged from 7 to 86%. By CR, deviations of average parameter estimates from the true parameter values were <0.10 log2 (mg/liter) for all parameters for each of the 26 pairs. By SR, these deviations were >0.10 log2 (mg/liter) for at least 18 of the 26 pairs for all but one parameter. Two-standard-error confidence intervals for individual parameters contained as little as 0% of cases for all SR approaches but > or = 91.5% of cases for the CR approach. When censored MIC data are modeled, CR may reduce or eliminate biased parameter estimates obtained by SR. PMID- 16377669 TI - Lack of pharmacokinetic interaction between linezolid and antacid in healthy volunteers. AB - Several antibiotics show significant pharmacokinetic interactions when they are given orally concomitantly with antacids. The objective of this study was to evaluate the effects of antacid (containing magnesium) on the pharmacokinetics of linezolid. A single dose of 600 mg linezolid was given orally alone and 10 min after administration of the antacid Maalox 70mVal, which contains 600 mg magnesium hydroxide and 900 mg aluminum hydroxide, to nine healthy males and nine healthy females in a crossover and randomized study. Linezolid plasma concentrations were determined by high-performance liquid chromatography, and pharmacokinetic parameters were calculated for both treatments. Coadministration with antacids did not change the pharmacokinetics of linezolid. The ratios (90% confidence intervals) of the individual values of the area under the concentration-time curve and the maximum concentration in plasma (C(max)) (linezolid plus antacid versus linezolid alone) were 1.01 (0.99 to 1.02) and 0.99 (0.96 to 1.02), respectively. Likewise, no significant difference in any of the other pharmacokinetic parameters was observed between the treatment groups (the time to C(max), lag time, volume of distribution [V/F], and clearance [CL/F]). However, a significant sex difference was observed for AUC, C(max), V/F, and CL/F; and these differences could be almost completely explained by the differences in body weight between males and females. No clinically relevant adverse effects were detected under either condition. The coadministration of antacids had no effect on the pharmacokinetics of linezolid. This demonstrates that the oral absorption of linezolid was not affected by the presence of antacids containing magnesium hydroxide and aluminum hydroxide. Antacids can be safely administered together with linezolid. PMID- 16377670 TI - In vitro and in vivo interactions between miltefosine and other antileishmanial drugs. AB - The interaction of miltefosine with amphotericin B, sodium stibogluconate, paromomycin, and sitamaquine was assessed in vitro and additionally for the first three combinations in vivo. In vitro interactions were indifferent for miltefosine combined with amphotericin B (mean sums of fractional inhibitory concentrations [mean summation operatorFICs] ranging from 1.22 to 1.51 at the 50% effective concentration [EC50] level and 1.08 to 1.38 at the EC90 level), sitamaquine (mean summation operatorFICs from 1.33 to 1.38 and 1.0 to 1.02, respectively), and paromomycin (mean summation operatorFICs from 0.79 to 0.93 at the EC50 and 0.77 to 1.35 at the EC90 level). Some synergy was observed for miltefosine combined with sodium stibogluconate (mean summation operatorFICs from 0.61 to 0.75 at EC50 and 0.49 to 0.97 at EC90). Different interactions were found in vivo, where the highest potentiation of miltefosine activity was achieved with amphotericin B (activity enhancement index [AEI] of up to 11.3). No significant interaction was observed when miltefosine was combined with sodium stibogluconate (AEI of up to 2.38). The potentiation of miltefosine in vivo was also achieved with the combination of miltefosine and paromomycin (AEI of up to 7.22). PMID- 16377671 TI - Involvement of the drug transporters p glycoprotein and multidrug resistance associated protein Mrp2 in telithromycin transport. AB - The present study aims to investigate the role of P glycoprotein and multidrug resistance-associated protein (Mrp2) in the transport of telithromycin, a newly developed ketolide antibiotic, in vitro and in vivo. The in vitro experiments revealed that the intracellular accumulation of telithromycin in adriamycin resistant human chronic myelogenous leukemia cells (K562/ADR) overexpressing P glycoprotein was significantly lower than that in human chronic myelogenous leukemia cells (K562/S) not expressing P glycoprotein. Cyclosporine significantly increased the intracellular accumulation of telithromycin in K562/ADR cells. When telithromycin was coadministered intravenously with cyclosporine in Sprague Dawley (SD) rats, cyclosporine significantly delayed the disappearance of telithromycin from plasma and decreased its systemic clearance to 60% of the corresponding control values. Hepatobiliary excretion experiments revealed that cyclosporine almost completely inhibited the biliary clearance of telithromycin, suggesting that telithromycin is a substrate of P glycoprotein and a potential substrate of Mrp2. Moreover, the biliary clearance of telithromycin was significantly decreased by 80% in Eisai hyperbilirubinemic mutant rats with a hereditary deficiency in Mrp2, indicating that Mrp2, as well as P glycoprotein, plays an important role in the biliary excretion of telithromycin. When the effect of telithromycin on the biliary excretion of doxorubicin, a substrate of P glycoprotein and Mrp2, was examined in SD rats, telithromycin significantly decreased the biliary clearance of doxorubicin by 80%. Results obtained from this study indicate that telithromycin is a substrate of both P glycoprotein and Mrp2, and these transporters are involved in the hepatobiliary transport of telithromycin. PMID- 16377672 TI - Resistance of Leishmania donovani to sodium stibogluconate is related to the expression of host and parasite gamma-glutamylcysteine synthetase. AB - Sequencing studies showed that the gamma-glutamylcysteine synthetase (gamma-GCS) heavy chain genes from sodium stibogluconate (SSG)-resistant (SSG-R) and SSG susceptible (SSG-S) Leishmania donovani strains were identical, indicating that SSG resistance was related to quantitative differences in gamma-GCS expression rather than gene interstrain polymorphisms. In vitro infection of murine macrophages with the SSG-R strain, but not the SSG-S strain, down regulated expression of host gamma-GCS, which would result in a reduction in intramacrophage glutathione (GSH) levels and promote an oxidative intramacrophage environment. This would inhibit, or minimize, the reduction of SSG pentavalent antimony to its more toxic trivalent form. Macrophage studies showed that the SSG R strain expressed higher levels of gamma-GCS compared to the SSG-S strain, which would result in higher GSH levels, giving increased protection against oxidative stress and facilitating SSG efflux. However a similar differential effect on host and parasite gamma-GCS expression was not obtained when using tissues from infected mice. In this case gamma-GCS expression was organ and strain dependent for both the host and the parasite, indicating that environmental conditions have a profound effect on gamma-GCS expression. Consistent with the proposed mechanism from in vitro studies, increasing tissue GSH levels in the presence of SSG by cotreatment of L. donovani-infected mice with SSG solution and GSH incorporated into nonionic surfactant vesicles was more effective in reducing liver, spleen, and bone marrow parasite burdens than monotherapy with SSG. Together, these results indicate that SSG resistance is associated with manipulation of both host and parasite GSH levels by L. donovani. PMID- 16377673 TI - Lovastatin has significant activity against zygomycetes and interacts synergistically with voriconazole. AB - Zygomycetes are emerging opportunistic molds resistant to most conventional antifungals. We evaluated the in vitro activity of lovastatin (LOV), a 3-hydroxy 3-methylglutaryl-coenzyme A reductase inhibitor, against seven clinical isolates of Zygomycetes by using standard microdilution methods in three different media, disk diffusion testing, and viability dye staining. To further study the in vivo efficacy of LOV against zygomycetes, we developed a Drosophila melanogaster model of zygomycosis. In different experiments, groups of Toll-deficient (Tl) flies fed LOV-containing food were subsequently injected with two representative Zygomycetes isolates (Mucor and Rhizopus spp.). Finally, we examined the effects of LOV on voriconazole (VRC) activity against zygomycetes in vitro by checkerboard dilution, Epsilometer test-based methods, and bis-(1,3 dibutylbarbituric acid) trimethine oxonol staining and in vivo in Tl flies fed food containing LOV plus VRC and infected with zygomycetes. LOV exhibited significant, medium, and strain-independent fungicidal activity against all Zygomycetes isolates in vitro by all testing methods (MIC50, 48.0 microg/ml; 50% minimal fungicidal concentration, 56.0 microg/ml; 50% effective concentration, 29.4 microg/ml [6.6 to 38.9 microg/ml]). Tl flies fed LOV-containing food and infected with Mucor had a significantly better 6-day survival rate than did infected Tl flies fed regular food (P = 0.0005). LOV displayed in vitro synergy with VRC against all Zygomycetes isolates (fractional inhibitory concentration index, 0.104 to 0.290) by all methods used. LOV also displayed synergy with VRC in the Drosophila model of zygomycosis (P < 0.01). LOV is significantly active against zygomycetes and synergizes with triazoles inherently resistant to them, such as VRC. The clinical significance of these findings needs to be further explored. PMID- 16377674 TI - Novel gyrase mutations in quinolone-resistant and -hypersusceptible clinical isolates of Mycobacterium tuberculosis: functional analysis of mutant enzymes. AB - Mutations in the DNA gyrase GyrA2GyrB2 complex are associated with resistance to quinolones in Mycobacterium tuberculosis. As fluoroquinolones are being used increasingly in the treatment of tuberculosis, we characterized several multidrug resistant clinical isolates of M. tuberculosis carrying mutations in the genes encoding the GyrA or GyrB subunits associated with quinolone resistance or hypersusceptibility. In addition to the reported putative quinolone resistance mutations in GyrA, i.e., A90V, D94G, and D94H, we found that the GyrB N510D mutation was also associated with ofloxacin resistance. Surprisingly, several isolates bearing a novel combination of gyrA T80A and A90G changes were hypersusceptible to ofloxacin. M. tuberculosis GyrA and GyrB subunits (wild type [WT] and mutants) were overexpressed in Escherichia coli, purified to homogeneity, and used to reconstitute highly active gyrase complexes. Mutant proteins were produced similarly from engineered gyrA and gyrB alleles by mutagenesis. MICs, enzyme inhibition, and drug-induced DNA cleavage were determined for moxifloxacin, gatifloxacin, ofloxacin, levofloxacin, and enoxacin. Mutant gyrase complexes bearing GyrA A90V, D94G, and D94H and GyrB N510D were resistant to quinolone inhibition (MICs and 50% inhibitory concentrations [IC50s] at least 3.5-fold higher than the concentrations for the WT), and all, except the GyrB mutant, were less efficiently trapped as a quinolone cleavage complex. In marked contrast, gyrase complexes bearing GyrA T80A or A90G were hypersusceptible to the action of many quinolones, an effect that was reinforced for complexes bearing both mutations (MICs and IC50s up to 14-fold lower than the values for the WT). This is the first detailed enzymatic analysis of hypersusceptibility and resistance in M. tuberculosis. PMID- 16377675 TI - Efficacy of amphotericin B in combination with flucytosine against flucytosine susceptible or flucytosine-resistant isolates of Cryptococcus neoformans during disseminated murine cryptococcosis. AB - Whether or not flucytosine should be administered to patients infected with Cryptococcus neoformans isolates found to be resistant to flucytosine in vitro remains a controversial issue. Thus, the efficacy of amphotericin B and flucytosine in combination was investigated by mortality and fungal burden studies in a murine model of disseminated cryptococcosis using two clinical isolates of Cryptococcus neoformans, one susceptible and one resistant (i.e., 64 microg/ml) to flucytosine. Amphotericin B was given intraperitoneally at 0.25 or 0.5 mg/kg/day, while flucytosine was given at 100 or 250 mg/kg/day orally. Treatment was started 24 h or day 6 after inoculation and continued for 5 days in fungal burden and mortality studies, respectively. The combination of amphotericin B at 0.5 mg/kg/day and flucytosine at 250 mg/kg/day was significantly more effective than monotherapies for reducing fungal burden in brain, spleen, and lungs after infection by the flucytosine-susceptible isolate and in brain and spleen for the flucytosine-resistant isolate. For the flucytosine-resistant isolate, the combination of amphotericin B at 0.5 mg/kg/day with flucytosine at 100 mg/kg/day was significantly better than monotherapies for reducing the fungal burden in the brain. Survival obtained after the combination of amphotericin B at 0.5 mg/kg/day and flucytosine at 250 mg/kg/day increased compared to that obtained with monotherapies for both isolates, but the difference was statistically significant only for the flucytosine-susceptible isolate. Antagonism was never observed. This study demonstrates the beneficial effect of the addition of flucytosine to amphotericin B against experimental disseminated cryptococcal infection even when the C. neoformans isolate is resistant to flucytosine. PMID- 16377676 TI - In vivo efficacies and pharmacokinetics of DX-619, a novel des-fluoro(6) quinolone, against Streptococcus pneumoniae in a mouse lung infection model. AB - DX-619 is a novel des-fluoro(6) quinolone with potent activity against gram positive pathogens. The in vivo activity of DX-619 against Streptococcus pneumoniae was compared with those of fluoro(6) quinolones, sitafloxacin, and ciprofloxacin in a mouse model. Two strains of S. pneumoniae were used: a penicillin-sensitive S. pneumoniae (PSSP) strain and a penicillin-resistant S. pneumoniae (PRSP) strain. Furthermore, these strains showed intermediate susceptibilities to ciprofloxacin. In murine lung infections caused by PSSP, the 50% effective doses (ED50s) of DX-619, sitafloxacin, and ciprofloxacin were 9.15, 11.1, and 127.6 mg/kg of body weight, respectively. Against PRSP-mediated pneumonia in mice, the ED50s of DX-619, sitafloxacin, and ciprofloxacin were 0.69, 4.84, and 38.75 mg/kg, respectively. The mean +/- standard error of the mean viable bacterial counts in murine lungs infected with PSSP and treated with DX-619, sitafloxacin, ciprofloxacin (10 mg/kg twice daily), and saline (twice daily) were 1.75 +/- 0.06, 1.92 +/- 0.23, 6.48 +/- 0.28, and 7.57 +/- 0.13 log10 CFU/ml, respectively. Furthermore, the numbers of viable bacteria in lungs infected with PRSP and treated with the three agents and not treated (control) were 1.73 +/- 0.04, 2.28 +/- 0.17, 4.61 +/- 0.59, and 5.54 +/- 0.72 log10 CFU/ml, respectively. DX-619 and sitafloxacin significantly decreased the numbers of viable bacteria in the lungs compared to the numbers in the lungs of ciprofloxacin-treated and untreated mice. The pharmacokinetic parameter of the area under the concentration-time curve (AUC)/MIC ratio in the lungs for DX-619, sitafloxacin, and ciprofloxacin were 171.0, 21.92, and 1.22, respectively. The AUC/MIC ratio in the lungs was significantly higher for DX-619 than for sitafloxacin and ciprofloxacin. Our results suggest that DX-619 and sitafloxacin are potent against both PSSP and PRSP in our mouse pneumonia model. PMID- 16377677 TI - Posaconazole as salvage therapy for zygomycosis. AB - Zygomycosis, an infection that is associated with significant morbidity and mortality, is becoming common in immunocompromised patients. Posaconazole is a new extended-spectrum azole antifungal that has demonstrated in vitro and in vivo activity against zygomycetes. This report provides the results from the first 24 patients with active zygomycosis who were enrolled in two open-label, nonrandomized, multicentered compassionate trials that evaluated oral posaconazole as salvage therapy for invasive fungal infections. Posaconazole was usually given as an oral suspension of 200 mg four times a day or 400 mg twice a day. Eleven (46%) of the infections were rhinocerebral. Duration of posaconazole therapy ranged from 8 to 1,004 days (mean, 292 days; median, 182 days). Rates of successful treatment (complete cure and partial response) were 79% in 19 subjects with zygomycosis refractory to standard therapy and 80% in 5 subjects with intolerance to standard therapy. Overall, 19 of 24 subjects (79%) survived infection. Survival was also associated with surgical resection of affected tissue and stabilization or improvement of the subjects' underlying illnesses. Failures either had worsening of underlying illnesses or requested all therapy withdrawn; none of the failures received more than 31 days of posaconazole. Posaconazole oral solution was well tolerated and was discontinued in only one subject due to a drug rash. Posaconazole appears promising as an oral therapy for zygomycosis in patients who receive required surgery and control their underlying illness. PMID- 16377678 TI - Preliminary mapping of a putative inhibitor-binding pocket for human immunodeficiency virus type 1 integrase inhibitors. AB - Molecular modeling studies have identified a putative human immunodeficiency virus (HIV) integrase (IN) inhibitor-binding pocket for l-chicoric acid (l-CA) and other inhibitors of IN (C. A. Sotriffer, H. Ni, and A. McCammon, J. Med. Chem. 43:4109-4117, 2000). By using site-directed mutagenesis of several amino acid residues identified by modeling studies, a common inhibitor-binding pocket on IN was confirmed for l-CA and the diketo acid L-731,988. Specifically, the single mutations E92K, Q148A, K156A, K156R, G140S, and G149S, as well as the double mutations C65S-K156N and H67D-G140A were evaluated for their effects on enzymatic activity and inhibitor susceptibility. Each recombinant IN was attenuated for 3'-end processing and strand transfer activities. Most proteins were also attenuated for disintegration; the IN that contained K156R and C65S K156N, however, displayed disintegration activity similar to that of IN from HIV(NL4-3). All mutant IN proteins demonstrated decreased susceptibility to l-CA, while all mutant proteins except E92K and K156R demonstrated resistance to L 731,988. These data validate the computer modeling data and demonstrate that l-CA and L-731,988 share an overlapping inhibitor-binding pocket that involves amino acids Q148, C65, and H67. The resistance studies confirm that L-731,988 fills one half of the inhibitor-binding pocket and binds to Q148 but excludes E92, while l CA fills the entire binding groove and thus interacts with E92. These results provide "wet laboratory" evidence that molecular models of the HIV IN inhibitor binding pocket can be used for drug discovery. PMID- 16377679 TI - Open-label, randomized comparison of itraconazole versus caspofungin for prophylaxis in patients with hematologic malignancies. AB - Invasive fungal infection remains the most common cause of infectious death in acute leukemia. In this open-label, randomized study, we compared the efficacy and safety of caspofungin with that of intravenous itraconazole for antifungal prophylaxis in patients undergoing induction chemotherapy for acute myelogenous leukemia or myelodysplastic syndrome. Of 200 patients, 192 were evaluable for efficacy (86 for itraconazole, 106 for caspofungin). Duration of prophylaxis (median, 21 days [range, 1 to 38 days]), demographics, and prognostic factors were similar in both groups. Ninety-nine patients completed antifungal prophylaxis without developing fungal infection (44 [51%] with itraconazole, 55 [52%] with caspofungin). Twelve patients developed documented invasive fungal infections, five in the itraconazole group (four with candidemia and one with Aspergillus pneumonia), and seven in the caspofungin group (two with candidemia, two with disseminated trichosporon species, two with Aspergillus pneumonia, and one with disseminated Fusarium spp). Two patients in the itraconazole group and four in the caspofungin group died of fungal infection (P = 0.57). Grade 3 to 4 adverse event rates were comparable between groups; the most common event in both was reversible hyperbilirubinemia. No evidence of cardiovascular toxicity from intravenous itraconazole was noted among patients older than 60. In conclusion, intravenous itraconazole and caspofungin provided similar protection against invasive fungal infection during induction chemotherapy, and both drugs were well tolerated. PMID- 16377680 TI - In vitro activity of moxifloxacin against 923 anaerobes isolated from human intra abdominal infections. AB - The in vitro activity of moxifloxacin against 923 recent anaerobic isolates obtained from pretreatment cultures in patients with complicated intra-abdominal infections was studied using the CLSI M11-A-6 agar dilution method. Moxifloxacin was active against 87% (96 of 110) Bacteroides fragilis strains at < or = 1 microg/ml and 87% (79 of 90) B. thetaiotaomicron strains at < or = 2 microg/ml. Species variation was seen, with B. uniformis, B. vulgatus, Clostridium clostridioforme, and C. symbiosum being least susceptible and accounting for most of the resistant isolates; excluding the aforementioned four resistant species, 86% (303 of 363) of Bacteroides species isolates and 94% (417 of 450) of all other genera and species were susceptible to < or = 2 microg/ml of moxifloxacin. Overall, moxifloxacin was active against 763 of 923 (83%) of strains at < or = 2 microg/ml, supporting its use as a monotherapy for some community-acquired intra abdominal infections. PMID- 16377681 TI - Tetracycline resistance in Escherichia coli and persistence in the infantile colonic microbiota. AB - The ecological impact of antibiotic resistance in the absence of selective pressure has been poorly studied. We assessed the carriage of tetracycline resistance genes, persistence in the microbiota, fecal population counts and virulence factor genes in 309 commensal, intestinal Escherichia coli strains obtained from 128 Swedish infants followed during the first year of life with regular quantitative fecal cultures. No infant was given tetracycline, but 25% received other antibiotics. Tetracycline resistance was identified in 12% of strains, all of which carried either tet(A) (49%) or tet(B) (51%) genes. Resistance to other antibiotics occurred in 50% of tet(A)-positive strains, 42% of tet(B)-positive strains and 13% of tetracycline-sensitive strains. However, colonization with tetracycline-resistant strains was unrelated to treatment with antibiotics. Strains that were tet(B)- or tet(A)-positive carried the genes for P fimbriae and aerobactin, respectively, more often than susceptible strains. Tetracycline-resistant and -susceptible strains were equally likely to persist among the intestinal microbiota for > or = 3 weeks and had similar population numbers. However, when a resistant strain and a susceptible strain colonized a child simultaneously, the resistant variety showed lower counts (P = 0.03). In cases of long-term colonization by initially tetracycline-resistant E. coli strains, loss of tet genes occurred in 3 of 13 cases with variable effects on population counts. The results indicate that there is limited pressure against the carriage of tet genes in the infantile gut microbiota even in the absence of antibiotics. Resistant strains may possess colonization factors that balance the cost of producing resistance elements. PMID- 16377682 TI - In vitro effects of thiazolides on Giardia lamblia WB clone C6 cultured axenically and in coculture with Caco2 cells. AB - The thiazolides represent a novel class of anti-infective drugs, with the nitrothiazole nitazoxanide [2-acetolyloxy-N-(5-nitro 2-thiazolyl) benzamide] (NTZ) as the parent compound. NTZ exhibits a broad spectrum of activities against a wide variety of helminths, protozoa, and enteric bacteria infecting animals and humans. In vivo, NTZ is rapidly deacetylated to tizoxanide (TIZ), which exhibits similar activities. We have here comparatively investigated the in vitro effects of NTZ, TIZ, a number of other modified thiazolides, and metronidazole (MTZ) on Giardia lamblia trophozoites grown under axenic culture conditions and in coculture with the human cancer colon cell line Caco2. The modifications of the thiazolides included, on one hand, the replacement of the nitro group on the thiazole ring with a bromide, and, on the other hand, the differential positioning of methyl groups on the benzene ring. Of seven compounds with a bromo instead of a nitro group, only one, RM4820, showed moderate inhibition of Giardia proliferation in axenic culture, but not in coculture with Caco2 cells, with a 50% inhibitory concentration (IC50) of 18.8 microM; in comparison, NTZ and tizoxanide had IC50s of 2.4 microM, and MTZ had an IC50 of 7.8 microM. Moreover, the methylation or carboxylation of the benzene ring at position 3 resulted in a significant decrease of activity, and methylation at position 5 completely abrogated the antiparasitic effect of the nitrothiazole compound. Trophozoites treated with NTZ showed distinct lesions on the ventral disk as soon as 2 to 3 h after treatment, whereas treatment with metronidazole resulted in severe damage to the dorsal surface membrane at later time points. PMID- 16377683 TI - Ciprofloxacin and trimethoprim cause phage induction and virulence modulation in Staphylococcus aureus. AB - In Staphylococcus aureus strains of human origin, phages which integrate into the chromosomal gene coding for beta-hemolysin (hlb) are widely distributed. Most of them encode accessory virulence determinants such as staphylokinase (sak) or enterotoxins. Here, we analyzed the effects of ciprofloxacin and trimethoprim on phage induction and expression of phage-encoded virulence factors by using isolates from patients with cystic fibrosis for which the induction of hlb converting phages was demonstrated in vivo (C. Goerke, S. Matias y Papenberg, S. Dasbach, K. Dietz, R. Ziebach, B. C. Kahl, and C. Wolz, J. Infect. Dis. 189:724 734, 2004) as well as a phi13 lysogen of phage-cured strain 8325-4. Treatment of lysogens with subinhibitory concentrations of either antibiotic resulted in (i) delysogenization of strains resembling the isolates picked up after chronic lung infection and (ii) replication of phages in the bacterial host in a dose dependent manner. Ciprofloxacin treatment resulted in enhanced recA transcription, indicating involvement of the SOS response in phage mobilization. Induction of phi13 was linked to elevated expression of the phage-encoded virulence gene sak, chiefly due to the activation of latent phage promoters. In summary, we could show the induction of hlb-converting phages and a subsequent virulence modulation of the host bacterium by ciprofloxacin and trimethoprim. PMID- 16377684 TI - Novel plasmid-mediated 16S rRNA methylase, RmtC, found in a proteus mirabilis isolate demonstrating extraordinary high-level resistance against various aminoglycosides. AB - Proteus mirabilis ARS68, which demonstrated a very high level of resistance to various aminoglycosides, was isolated in 2003 from an inpatient in Japan. The aminoglycoside resistance of this strain could not be transferred to recipient strains Escherichia coli CSH-2 and E. coli HB101 by a general conjugation experiment, but E. coli DH5alpha was successfully transformed by electroporation with the plasmid of the parent strain, ARS68, and acquired an unusually high degree of resistance against aminoglycosides. Cloning and sequencing analyses revealed that the presence of a novel 16S rRNA methylase gene, designated rmtC, was responsible for resistance in strain ARS68 and its transformant. The G+C content of rmtC was 41.1%, and the deduced amino acid sequences of the newly identified 16S rRNA methylase, RmtC, shared a relatively low level of identity (< or = 29%) to other plasmid-mediated 16S rRNA methylases, RmtA, RmtB, and ArmA, which have also been identified in pathogenic gram-negative bacilli. Also, RmtC shared a low level of identity (< or = 28%) with the other 16S rRNA methylases found in aminoglycoside-producing actinomycetes. The purified histidine-tagged RmtC clearly showed methyltransferase activity against E. coli 16S rRNA in vitro. rmtC was located downstream of an ISEcp1-like element containing tnpA. Several plasmid-mediated 16S rRNA methylases have been identified in pathogenic gram negative bacilli belonging to the family Enterobacteriaceae, and some of them are dispersing worldwide. The acceleration of aminoglycoside resistance among gram negative bacilli by producing plasmid-mediated 16S rRNA methylases, such as RmtC, RmtB, and RmtA, may indeed become an actual clinical hazard in the near future. PMID- 16377685 TI - Two cathelicidin genes are present in both rainbow trout (Oncorhynchus mykiss) and atlantic salmon (Salmo salar). AB - Further to the previous finding of the rainbow trout rtCATH_1 gene, this paper describes three more cathelicidin genes found in salmonids: two in Atlantic salmon, named asCATH_1 and asCATH_2, and one in rainbow trout, named rtCATH_2. All the three new salmonid cathelicidin genes share the common characteristics of mammalian cathelicidin genes, such as consisting of four exons and possessing a highly conserved preproregion and four invariant cysteines clustered in the C terminal region of the cathelin-like domain. The asCATH_1 gene is homologous to the rainbow trout rtCATH_1 gene, in that it possesses three repeat motifs of TGGGGGTGGC in exon IV and two cysteine residues in the predicted mature peptide, while the asCATH_2 gene and rtCATH_2 gene are homologues of each other, with 96% nucleotide identity. Salmonid cathelicidins possess the same elastase-sensitive residue, threonine, as hagfish cathelicidins and the rabbit CAP18 molecule. The cleavage site of the four salmonid cathelicidins is within a conserved amino acid motif of QKIRTRR, which is at the beginning of the sequence encoded by exon IV. Two 36-residue peptides corresponding to the core part of rtCATH_1 and rtCATH_2 were chemically synthesized and shown to exhibit potent antimicrobial activity. rtCATH_2 was expressed constitutively in gill, head kidney, intestine, skin and spleen, while the expression of rtCATH_1 was inducible in gill, head kidney, and spleen after bacterial challenge. Four cathelicidin genes have now been characterized in salmonids and two were identified in hagfish, confirming that cathelicidin genes evolved early and are likely present in all vertebrates. PMID- 16377686 TI - Phenothiazinium antimicrobial photosensitizers are substrates of bacterial multidrug resistance pumps. AB - Antimicrobial photodynamic therapy (PDT) combines a nontoxic photoactivatable dye, or photosensitizer (PS), with harmless visible light to generate singlet oxygen and free radicals that kill microbial cells. Although the light can be focused on the diseased area, the best selectivity is achieved by choosing a PS that binds and penetrates microbial cells. Cationic phenothiazinium dyes, such as methylene blue and toluidine blue O, have been studied for many years and are the only PSs used clinically for antimicrobial PDT. Multidrug resistance pumps (MDRs) are membrane-localized proteins that pump drugs out of cells and have been identified for a wide range of organisms. We asked whether phenothiazinium salts with structures that are amphipathic cations could potentially be substrates of MDRs. We used MDR-deficient mutants of Staphylococcus aureus (NorA), Escherichia coli (TolC), and Pseudomonas aeruginosa (MexAB) and found 2 to 4 logs more killing than seen with wild-type strains by use of three different phenothiazinium PSs and red light. Mutants that overexpress MDRs were protected from killing compared to the wild type. Effective antimicrobial PSs of different chemical structures showed no difference in light-mediated killing depending on MDR phenotype. Differences in uptake of phenothiazinium PS by the cells depending on level of MDR expression were found. We propose that specific MDR inhibitors could be used in combination with phenothiazinium salts to enhance their photodestructive efficiency. PMID- 16377687 TI - Simultaneous detection of nine antibiotic resistance-related genes in Streptococcus agalactiae using multiplex PCR and reverse line blot hybridization assay. AB - Streptococcus agalactiae (group B streptococcus [GBS]) is the leading cause of neonatal and maternal sepsis. Penicillin is recommended for intrapartum prophylaxis, but erythromycin or clindamycin is used for penicillin-allergic carriers. Antibiotic resistance (AR) has increased recently and needs to be monitored. We have developed a multiplex PCR-based reverse line blot (mPCR/RLB) hybridization assay to detect, simultaneously, seven genes encoding AR- erm(A/TR), erm(B), mef(A/E), tet(M), tet(O), aphA-3, and aad-6--and two AR related genes, int-Tn and mreA. We tested 512 GBS isolates from Asia and Australasia and compared mPCR/RLB with antibiotic susceptibility phenotype or single-gene PCR. Phenotypic resistance to tetracycline was identified in 450 (88%) isolates, of which 442 had tet(M) (93%) and/or tet(O) (6%). Of 67 (13%) erythromycin-resistant isolates, 18 were susceptible to clindamycin, i.e., had the M phenotype, encoded by mef(A/E); 39 had constitutive (cMLS(B)) and 10 inducible clindamycin resistance, and of these, 34 contained erm(B) and 12 erm(A/TR). Of four additional isolates with mef(A/E), three contained erm(B) with cMLS(B) and one was erythromycin susceptible. Of 61 (12%) clindamycin-resistant isolates, 20 were susceptible to erythromycin and two had intermediate resistance. Based on sequencing, 21 of 22 isolates with mef had mef(E), and 8 of 353 with int-Tn had an atypical sequence. Several AR genes, erm(B), tet(O), aphA 3, aad-6, and mef(A/E), were significantly more common among Asian than Australasian isolates, and there were significant differences in distribution of AR genes between GBS serotypes. Our mPCR/RLB assay is simple, rapid, and suitable for surveillance of antibiotic resistance in GBS. PMID- 16377688 TI - Effect of ethanol on fluoroquinolone efficacy in a rat model of pneumococcal pneumonia. AB - This investigation compared the effect of ethanol on fluoroquinolone antibiotic efficacy and pharmacodynamics in an ethanol-fed rat model of pneumococcal pneumonia. Male Sprague-Dawley rats received a liquid diet containing 36% of total calories as ethanol. Paired controls (pair-fed controls) were fed a liquid diet without ethanol or received rat chow. Diets began 7 days before and continued for 10 days after transtracheal infections with 10 times the 50% lethal dose of type 3 Streptococcus pneumoniae. Beginning 18 h after infection, the rats received once daily subcutaneous phosphate-buffered saline, levofloxacin, moxifloxacin, or trovafloxacin at 50 or 100 mg/kg of body weight. White blood cell counts were determined, blood samples were collected for culture, and mortality was recorded. Additional rats were killed on day 5 for pharmacodynamic studies and quantitative cultures of bronchoalveolar lavage fluid. Bacteremia occurred by day 3 in 20 of 22 untreated rats. All 22 untreated rats died by day 9. Moxifloxacin treatment was effective in all diet groups at both the 50- and 100-mg/kg doses. In contrast, 50-mg/kg doses of levofloxacin and trovafloxacin improved survival in ethanol-fed rats but were ineffective in chow-fed rats. High dose trovafloxacin at 100 mg/kg was associated with increased mortality in pair fed rats. The free-fraction area under the concentration-time curve/MIC ratio exceeded 50 with all antibiotics in the ethanol group but dropped below 30 with levofloxacin and trovafloxacin in the pair- and chow-fed rats, with higher mortality. Achievement of adequate antibiotic-free fraction area under the concentration-time curve/MIC ratios helps overcome ethanol-induced immune defects induced in experimental pneumococcal pneumonia. PMID- 16377689 TI - Induction and inhibition of ciprofloxacin resistance-conferring mutations in hypermutator bacteria. AB - The emergence of drug-resistant bacteria poses a serious threat to human health. Bacteria often acquire resistance from a mutation of chromosomal genes during therapy. We have recently shown that the evolution of resistance to ciprofloxacin in vivo and in vitro requires the induction of a mutation that is mediated by the cleavage of the SOS repressor LexA and the associated derepression of three specialized DNA polymerases (polymerase II [Pol II], Pol IV, and Pol V). These results led us to suggest that it may be possible to design drugs to inhibit these proteins and that such drugs might be coadministered with antibiotics to prevent mutation and the evolution of resistance. For the approach to be feasible, there must not be any mechanisms through which bacteria can induce mutations and acquire antibiotic resistance that are independent of LexA and its repressed polymerases. Perhaps the most commonly cited mechanism to elevate bacterial mutation rates is the inactivation of methyl-directed mismatch repair (MMR). However, it is unclear whether this represents a LexA-independent mechanism or if the mutations that arise in MMR-deficient hypermutator strains are also dependent on LexA cleavage and polymerase derepression. In this work, we show that LexA cleavage and polymerase derepression are required for the evolution of clinically significant resistance in MMR-defective Escherichia coli. Thus, drugs that inhibit the proteins responsible for induced mutations are expected to efficiently prevent the evolution of resistance, even in MMR deficient hypermutator strains. PMID- 16377690 TI - First detection of metallo-beta-lactamase VIM-2 in Pseudomonas aeruginosa isolates from Colombia. AB - Carbapenem resistance rates in Pseudomonas aeruginosa isolates in Colombia, as in many South American countries, are high for reasons that remain unclear. From our nationwide network, we describe the first detection of the metallo-beta-lactamase VIM-2 in clinical isolates of P. aeruginosa from multiple cities within Colombia. Metallo-beta-lactamases were not detected in the two centers with the highest imipenem resistance rates. Clonality was noted in five of the eight centers with strains meeting the criteria for molecular typing. The high carbapenem resistance in P. aeruginosa in Colombia may be attributable to a combination of factors, including the presence of metallo-beta-lactamases and nosocomial transmission. PMID- 16377691 TI - Utility of muropeptide ligase for identification of inhibitors of the cell wall biosynthesis enzyme MurF. AB - MurF is a key enzyme in the biosynthesis of the bacterial cell wall in both gram positive and gram-negative bacteria. This enzyme has not been extensively exploited as a drug target, possibly due to the difficulty in obtaining one of the substrates, UDP-MurNAc-L-Ala-gamma-D-Glu-meso-diaminopimelate, which is usually purified from bacteria. We have identified putative inhibitors of Escherichia coli MurF by a binding assay, thus bypassing the need for substrate. Inhibition of enzymatic activity was demonstrated in a high-performance liquid chromatography-based secondary assay with UDP-MurNAc-L-Ala-gamma-D-Glu diaminopimelate substrate prepared in a novel way by using muropeptide ligase enzyme to add UDP-MurNAc to synthetic L-Ala-gamma-D-Glu-diaminopimelate; the substrate specificity of muropeptide ligase for peptides containing L-Lys in place of diaminopimelate was also investigated. Using the muropeptide ligase generated MurF substrate, a thiazolylaminopyrimidine series of MurF enzyme inhibitors with 50% inhibitory concentration values as low as 2.5 microM was identified. PMID- 16377692 TI - Activity of a new oral streptogramin, XRP2868, against gram-positive cocci harboring various mechanisms of resistance to streptogramins. AB - The antibacterial activity of XRP2868, a new oral streptogramin composed of a combination of RPR132552 (streptogramin A) and RPR202868 (streptogramin B), was evaluated against a collection of clinical gram-positive isolates with characterized phenotypes and genotypes of streptogramin resistance. The effects of genes for resistance to streptogramin A or B on the activity of XRP2868 and its components were also tested by cloning these genes individually or in various combinations in gram-positive recipient strains susceptible to quinupristin dalfopristin. The species tested included Staphylococcus aureus, coagulase negative staphylococci, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, and other species of streptococci. XRP2868 was generally fourfold more potent than quinupristin-dalfopristin against S. aureus, E. faecium, and streptococci and had activity against E. faecalis (MICs = 0.25 to 1 microg/ml). XRP2868 appeared to be affected by the same mechanisms of resistance as those to quinupristin-dalfopristin. Nevertheless, the strong activity of factor A of the oral streptogramin enabled the combination to be very potent against streptogramin-susceptible staphylococci, streptococci, and E. faecium (MICs = 0.03 to 0.25 microg/ml) and to retain low MICs against the strains harboring a mechanism of resistance to factor A or factor B of the streptogramin. However, the combination of mechanisms of resistance to factors A and B caused an increase in the MICs of XRP2868, which reached 1 to 4 mug/ml. As with the other streptogramins, there was a reduction in the bactericidal effect of XRPR2868 when the staphylococcal strains acquired a constitutively expressed erm gene. PMID- 16377693 TI - Pharmacodynamics of a new streptogramin, XRP 2868, in murine thigh and lung infection models. AB - XRP 2868 is a new streptogramin antibiotic with broad-spectrum activity against gram-positive cocci. We used the neutropenic murine thigh and lung infection models to characterize the time course of antimicrobial activity of XRP 2868 and determine which pharmacokinetic/pharmacodynamic (PK/PD) parameter and magnitude best correlated with efficacy. Serum levels following four two- to fourfold escalating single-dose levels of XRP 2868 were measured by liquid chromatography mass spectrometry assay. In vivo postantibiotic effects (PAEs) were determined after doses of 2.5, 10, and 40 mg/kg. Mice had 10(6.8) to 10(8.4) CFU/thigh of strains of Streptococcus pneumoniae ATCC 10813 or Staphylococcus aureus ATCC 29213 at the start of therapy when treated for 24 h with 2.5 to 640 mg/kg/day of XRP 2868 fractionated for 3-, 6-, 12-, and 24-h dosing regimens. Nonlinear regression analysis was used to determine which PK/PD parameter best correlated with CFU/thigh at 24 h. Pharmacokinetic studies exhibited peak dose values of 0.03 to 0.07, area under the concentration-time curve (AUC) dose values of 0.02 to 0.07, and half-lives of 0.35 to 1.27 h. XRP 2868 produced in vivo PAEs of 0.5 to 3.4 h with S. pneumoniae strain ATCC 10813 and -1.5 to 10.7 h with S. aureus strain ATCC 29213. The 24-h AUC/MIC was the PK/PD parameter that best correlated with efficacy. In subsequent studies, we used the neutropenic murine thigh infection model to determine if the magnitude of the AUC/MIC needed for the efficacy of XRP 2868 varied among pathogens (including resistant strains). Mice had 10(6.1) to 10(7.8) CFU/thigh of four isolates of S. aureus (three methicillin susceptible and one methicillin-resistant strain) and nine isolates of S. pneumoniae (one penicillin-susceptible, four penicillin-intermediate, and four penicillin-resistant strains) when treated for 24 h with 0.16 to 640 mg/kg of XRP 2868 every 6 h. A sigmoid dose-response model was used to estimate the doses (mg/kg/24 h) required to achieve a net bacteriostatic affect over 24 h. MICs ranged from 0.06 to 0.25 microg/ml. The 24-h AUC/MICs for each static dose (20.7 to 252 mg/kg/day) varied from 3 to 70. Mean 24-h AUC/MICs +/- standard deviations (SDs) for S. pneumoniae and S. aureus isolates were 14 +/- 10 and 31 +/- 16, respectively. Beta-lactam and macrolide resistance did not alter the magnitude of AUC/MIC required for efficacy. PMID- 16377694 TI - AsnB is involved in natural resistance of Mycobacterium smegmatis to multiple drugs. AB - Mycobacteria are naturally resistant to most common antibiotics and chemotherapeutic agents. The underlying molecular mechanisms are not fully understood. In this paper, we describe a hypersensitive mutant of Mycobacterium smegmatis, MS 2-39, which was isolated by screening for transposon insertion mutants of M. smegmatis mc2155 that exhibit increased sensitivity to rifampin, erythromycin, or novobiocin. The mutant MS 2-39 exhibited increased sensitivity to all three of the above mentioned antibiotics as well as fusidic acid, but its sensitivity to other antibiotics, including isoniazid, ethambutol, streptomycin, chloramphenicol, norfloxacin, tetracycline, and beta-lactams, remained unchanged. Uptake experiment with hydrophobic agents and cell wall lipid analysis suggest that the mutant cell wall is normal. The transposon insertion was localized within the asnB gene, which is predicted to encode a glutamine-dependent asparagine synthetase. Transformation of the mutant with wild-type asnB of mc2155 or asnB of Mycobacterium tuberculosis complemented the drug sensitivity phenotype. These results suggest that AsnB plays a role in the natural resistance of mycobacteria. PMID- 16377695 TI - Resistance to erythromycin and telithromycin in Streptococcus pyogenes isolates obtained between 1999 and 2002 from Greek children with tonsillopharyngitis: phenotypic and genotypic analysis. AB - Since the late 1990s, the prevalence of erythromycin-resistant Streptococcus pyogenes has significantly increased in several European countries. Between January 1999 and December 2002, 1,577 isolates of S. pyogenes were recovered from children with tonsillopharyngitis living in various areas of Western Greece. Erythromycin resistance was observed in 379 (24%) of the 1,577 isolates. All erythromycin-resistant strains along with 153 randomly selected erythromycin susceptible S. pyogenes isolates were tested for their antimicrobial susceptibility, resistance phenotypes, and genotypes. Representative isolates underwent emm gene sequence typing. Isolates with reduced susceptibility to telithromycin (MIC, > or = 2 microg/ml) were studied for multilocus sequence type, L22, L4, and 23S rRNA mutations. Of the total 379 erythromycin-resistant isolates, 193 (50.9%) harbored the mef(A) gene, 163 (43%) erm(A), 1 (0.3%) mef(A) plus erm(A), and 22 (5.8%) the erm(B) gene. Among the erythromycin-susceptible isolates, emm 1 (25%), emm 2 (12.5%), and emm 77 (12.5%) predominated. Furthermore, among the erythromycin-resistant isolates, emm 4 (30.6%), emm 28 (22.2%), and emm 77 (12.5%) prevailed. Resistance to telithromycin was observed in 22 (5.8%) of the erythromycin-resistant isolates. Sixteen (72.7%) of the 22 isolates appeared to be clonally related, since all of them belonged to emm type 28 and multilocus sequence type 52. One of the well-known mutations (T2166C) in 23S rRNA, as well as a new one (T2136C), was detected in erythromycin- and telithromycin-resistant isolates. High incidence of macrolide resistance and clonal spread of telithromycin resistance were the characteristics of the Greek S. pyogenes isolates obtained from 1999 to 2002. PMID- 16377696 TI - Topical voriconazole as a novel treatment for fungal keratitis. AB - Paecilomyces lilacinus is a fungal pathogen which is generally resistant to amphotericin B and certain other antifungals and is an uncommon cause of devastating fungal keratitis. In the present studies, we evaluated topical voriconazole as therapy for P. lilacinus keratitis in rabbits. Thirty eyes of 15 rabbits were studied. In five animals, the uninfected left eye was treated twice daily with voriconazole (drug control, uninfected eye). In these same animals, the right eye was infected with P. lilacinus but not treated with voriconazole (infection control eye). By day 5, the infection controls had lesions of >2.4 mm in diameter, with conjunctivitis and severe hypopyon, and were sacrificed. In the other 10 rabbits (voriconazole treatment), the right eyes were infected with P. lilacinus and treated with voriconazole beginning on day 3 after infection. Voriconazole therapy caused lesions to decrease during 8 days of therapy, after which rabbits were sacrificed (11 days postinfection). Hyphal masses were present in the control infected eyes and absent in treated infected eyes. Voriconazole was detected in all tissues of treated eyes. Topical voriconazole is effective treatment for P. lilacinus experimental keratitis, and it penetrates more deeply than the corneal tissue. PMID- 16377697 TI - Global transcriptome analysis of the responses of a fluoroquinolone-resistant Streptococcus pneumoniae mutant and its parent to ciprofloxacin. AB - Streptococcus pneumoniae M22 is a multidrug-resistant mutant selected after exposure of capsulated wild-type S. pneumoniae NCTC 7465 (strain M4) to ciprofloxacin. DNA microarray analysis comparing the gene expression profiles of strain M22 with those of strain M4 showed that strain M22 constitutively expressed 22 genes at levels higher than those observed in strain M4 under all conditions studied. These included the genes encoding the enzymes involved in branched-chain amino acid biosynthesis and two genes (patA and patB) with sequences suggestive of ABC transporter proteins. Expression of the patA and patB genes was induced by ciprofloxacin in both strains, but in strain M4 it only reached the levels observed in strain M22 after long incubation with high concentrations of ciprofloxacin. The altered expression profile observed with strain M22 suggested that the mutation or mutations acquired during resistance selection bring the cell into a state in which the expression of critical genes is preemptively altered to correct for the potential effects of ciprofloxacin on gene expression in the parent strain. PMID- 16377698 TI - Single-ascending-dose pharmacokinetics and safety of the novel broad-spectrum antifungal triazole BAL4815 after intravenous infusions (50, 100, and 200 milligrams) and oral administrations (100, 200, and 400 milligrams) of its prodrug, BAL8557, in healthy volunteers. AB - BAL8557 is the water-soluble prodrug of a novel antifungal triazole, BAL4815. BAL4815 is active against a broad spectrum of major opportunistic and pathogenic fungi, including strains that are resistant to other azoles. Cohorts of healthy male subjects received single-ascending oral (p.o.) doses of BAL8557 that were equivalent to 100, 200, or 400 mg of BAL4815 or single-ascending, 1-h constant rate intravenous (i.v.) infusions of BAL8557 which were equivalent to 50, 100, or 200 mg of BAL4815. In each cohort, six subjects were randomly assigned to receive active drug and two subjects were assigned to receive the placebo. All doses were well tolerated, and no severe or serious adverse events occurred. Maximum plasma concentrations of BAL4815 were observed 1.5 to 3 h after p.o. drug intake or at the end of the 1-h infusion. After both routes of administration, values for maximum drug concentration observed in plasma and area under the concentration time curve increased slightly more than proportionally to the administered dose. Mean elimination half-lives were particularly long (56 to 77 h after p.o. administration and 76 to 104 h after i.v. administration). The volume of distribution was large (155 to 292 liters after p.o. administration and 304 to 494 liters after i.v. administration) and systemic clearance was low (1.9 to 2.8 liter/h after p.o. administration and 2.8 to 5.0 liter/h after i.v. administration). Urinary recovery of BAL4815 was less than 0.4% of the infused dose. Based on the exposure data, oral bioavailability of BAL4815 is assumed to be very high. The pharmacokinetics of BAL4815 are well suited to maintaining concentrations of BAL4815 for a long period of time in the body and to enabling an effective treatment of systemic mycoses. PMID- 16377699 TI - Multiple-dose pharmacokinetics and safety of the new antifungal triazole BAL4815 after intravenous infusion and oral administration of its prodrug, BAL8557, in healthy volunteers. AB - BAL8557 is the water-soluble prodrug of BAL4815, a new broad-spectrum antifungal. Healthy male subjects were randomly assigned to four treatment cohorts to receive multiple oral doses or multiple 1-h constant-rate intravenous infusions of BAL8557. Loading doses of BAL8557 were equivalent to 100 mg (followed by once daily maintenance doses of 50 mg) or 200 mg (followed by once-daily maintenance doses of 100 mg) of BAL4815. In each cohort, six subjects received active drug and two subjects received the placebo. Study duration was 21 days (oral) and 14 days (intravenous). All adverse events reported were mild or moderate, except one severe rhinitis event which was not related to trial medication. After both routes of administration, maximum drug concentration observed in plasma (C(max)) and area under the concentration-time curve (AUC) values of BAL4815 increased proportionally to the administered dose. AUC values reflected a fourfold to fivefold accumulation of active drug in plasma during once-daily dosing, which is in line with the long elimination half-life of BAL4815 determined after the last administration (mean, 84.5 to 117 h). At steady state, the volume of distribution was large and amounted to 308 to 542 liters. Systemic clearance reached only 2.4 to 4.1 liter/h. At the levels obtained in the present study, C(max) values of 2.56 and 2.55 microg/ml after oral and intravenous administrations, respectively, there was no indication of CYP3A4 induction or inhibition (as revealed by the urinary 6-beta-hydroxycortisol/cortisol test). Based on AUC values after oral and intravenous administration, an excellent oral bioavailability can be predicted for BAL4815. Once-daily oral dosing of 50- or 100-mg equivalents of BAL8557 were recently demonstrated to be efficacious in a phase 2 study conducted with patients with esophageal candidiasis. These doses (preceded by adequate loading dose[s]) will be further explored in the treatment of systemic mycoses. PMID- 16377700 TI - Pentamidine is active in a neutropenic murine model of acute invasive pulmonary fusariosis. AB - We studied the efficacy of pentamidine (PNT) as prophylaxis or early treatment in acute pulmonary fusariosis in neutropenic mice. PNT-preexposed mice had significantly improved survival and reduced fungal burden compared to amphotericin B-preexposed and untreated mice. PNT-treated mice had increased survival but no difference in fungal burden versus untreated mice. PMID- 16377701 TI - Molecular genetic and structural modeling studies of Staphylococcus aureus RNA polymerase and the fitness of rifampin resistance genotypes in relation to clinical prevalence. AB - The adaptive and further evolutionary responses of Staphylococcus aureus to selection pressure with the antibiotic rifampin have not been explored in detail. We now present a detailed analysis of these systems. The use of rifampin for the chemotherapy of infections caused by S. aureus has resulted in the selection of mutants with alterations within the beta subunit of the target enzyme, RNA polymerase. Using a new collection of strains, we have identified numerous novel mutations in the beta subunits of both clinical and in vitro-derived resistant strains and established that additional, undefined mechanisms contribute to expression of rifampin resistance in clinical isolates of S. aureus. The fitness costs associated with rifampin resistance genotypes were found to have a significant influence on their clinical prevalence, with the most common clinical genotype (H481N, S529L) exhibiting no fitness cost in vitro. Intragenic mutations which compensate for the fitness costs associated with rifampin resistance in clinical strains of S. aureus were identified for the first time. Structural explanations for rifampin resistance and the loss of fitness were obtained by molecular modeling of mutated RNA polymerase enzymes. PMID- 16377702 TI - Quinolone efflux pumps play a central role in emergence of fluoroquinolone resistance in Streptococcus pneumoniae. AB - The preferential use of older antimicrobial agents is, in general, sound public health policy and is meant to maintain susceptibility to newer agents. In the case of fluoroquinolones, however, this strategy is flawed and may actually hasten the spread of Streptococcus pneumoniae strains resistant to newer members of the class. In a mouse thigh infection model, we were unable to isolate clones of pneumococci resistant to the newer fluoroquinolone levofloxacin at 2 x or 4 x the baseline MIC. An initial exposure in vivo to the older agent, ciprofloxacin, allowed straightforward selection of clones resistant to levofloxacin in a subsequent experiment. The original ciprofloxacin exposure generated clones without changes in the parC/E and gyrA/B quinolone target sites almost exclusively but did allow overexpression of a reserpine-responsive pump. While this caused only minimal change in the levofloxacin MIC (0.6 mg/liter to 0.8 mg/liter), it allowed a major change in the mutational frequency to resistance for levofloxacin (<1/10(8.5) to approximately 1/10(4.5)), which allowed levofloxacin-resistant clones to be isolated in a subsequent in vivo experiment. The reason underlying ciprofloxacin's propensity to select for pump-overexpressed clones is likely related to its hydrophilicity. To preserve the susceptibility of Streptococcus pneumoniae to newer members of the class of quinolones, use of ciprofloxacin for community-acquired respiratory infections should be minimized. PMID- 16377703 TI - Computer-aided design of agents that inhibit the cep quorum-sensing system of Burkholderia cenocepacia. AB - Recent research has provided evidence that interference with bacterial cell-to cell signaling is a promising strategy for the development of novel antimicrobial agents. Here we report on the computer-aided design of novel compounds that specifically inhibit an N-acyl-homoserine lactone-dependent communication system that is widespread among members of the genus Burkholderia. This genus comprises more than 30 species, many of which are important pathogens of animals and humans. Over the past few years, several Burkholderia species, most notably Burkholderia cenocepacia, have emerged as important opportunistic pathogens causing severe pulmonary deterioration in persons with cystic fibrosis. As efficient treatment of Burkholderia infections is hampered by the inherent resistance of the organisms to a large range of antibiotics, novel strategies for battling these pathogens need to be developed. Here we show that compounds targeting the B. cenocepacia signaling system efficiently inhibit the expression of virulence factors and attenuate the pathogenicity of the organism. PMID- 16377704 TI - Distinct antifungal mechanisms: beta-defensins require Candida albicans Ssa1 protein, while Trk1p mediates activity of cysteine-free cationic peptides. AB - Salivary histatin 5 (Hst 5) kills the fungal pathogen Candida albicans via a multistep process which includes binding to Ssa1/2 proteins on the cell surface and requires the TRK1 potassium transporter. Hst 5-induced membrane permeability to propidium iodide (PI) was nearly abolished in strain CaTK1 (TRK1/trk1), suggesting that Hst 5-induced influx of PI is via Trk1p. To explore the functional role of Trk1p in the mechanism of other antifungal peptides, we evaluated candidacidal activity and PI uptake in wild-type strain CaTK2 (TRK1/TRK1) and strain CaTK1 following treatment with lactoferricin 11 (LFcn 11), bactenecin 16 (BN 16), and virion-associated protein VPR 12. Strain CaTK1 was resistant to killing with these peptides (VPR 12 > LFcn 11 > BN 16), showing the requirement of Trk1p for fungicidal activity. In contrast, human neutrophil defensin 1 (HNP-1), human beta-defensin 2 (hBD-2), and hBD-3 effects on viability of and membrane permeability to PI were not different between mutant and wild type strains, clearly showing that their candidacidal mechanism does not involve Trk1p as a functional effector. To test whether defensins require binding to Candida surface Ssa1/2 proteins for their activity, we measured the killing effectiveness in SSA1/2 mutant strains. Both hBD-2 and hBD-3, but not HNP-1, exhibited reduced killing of ssa1Delta and ssa2Delta strains compared to the wild type, showing that Ssa1 and Ssa2 proteins are required for their fungicidal activity. These results demonstrate that (i) Trk1p mediates candidacidal activities of cysteine-free peptides, but not of defensins, and (ii) hBD-2 and hBD-3, but not HNP-1, require Ssa1/2p for antifungal activity. PMID- 16377705 TI - In vivo and in vitro studies of antibiotic release from and bacterial growth inhibition by antibiotic-impregnated polymethylmethacrylate hip spacers. AB - The antimicrobial properties and the elution characteristics of gentamicin vancomycin-loaded hip spacers were studied in vivo and in vitro. Vancomycin elution was greater than gentamicin elution. The antibiotic concentrations in vivo were less than those in vitro. Not dependent on implantation duration, growth inhibition by spacers in vitro was observed for 2 weeks. The reason for protracted wound healing cannot be insufficient antibiotic release. PMID- 16377706 TI - VraSR two-component regulatory system and its role in induction of pbp2 and vraSR expression by cell wall antimicrobials in Staphylococcus aureus. AB - The VraS/VraR two-component system (VraSR) regulates transcriptional induction of penicillin-binding protein 2 (encoded by pbp2) by vancomycin in Staphylococcus aureus. We have now defined the vraSR operon and determined that its induction by beta-lactams as well as by vancomycin is autoregulated. Induction of the pbp2 and vraSR operons by beta-lactams and related compounds within 1 hour after exposure to the antimicrobials was dependent on vraS. However, when a disk diffusion assay that can detect induction of genes over an extended time period was used, induction of the pbp2 operon was mediated by some beta-lactams, including oxacillin; this induction was independent of vraS. PMID- 16377707 TI - 5-Nitroimidazole drugs effective against metronidazole-resistant Trichomonas vaginalis and Giardia duodenalis. AB - Metronidazole (Mz)-resistant Giardia and Trichomonas were inhibited by 1 of 30 new 5-nitroimidazole drugs. Another five drugs were effective against some but not all of the Mz-resistant parasites. This study provides the incentive for the continued design of 5-nitroimidazole drugs to bypass cross-resistance among established 5-nitromidazole antiparasitic drugs. PMID- 16377708 TI - Anucleate cell blue assay: a useful tool for identifying novel type II topoisomerase inhibitors. AB - About 95,000 compounds were screened by the anucleate cell blue assay. Fifty-one of the hit compounds had various structures and showed inhibitory activity against DNA gyrase and/or topoisomerase IV. Moreover, the compounds exhibited antibacterial activity against a fluoroquinolone- and novobiocin-resistant strain of Staphylococcus aureus. The anucleate cell blue assay is therefore a useful tool for finding novel type II topoisomerase inhibitors. PMID- 16377709 TI - The K101P and K103R/V179D mutations in human immunodeficiency virus type 1 reverse transcriptase confer resistance to nonnucleoside reverse transcriptase inhibitors. AB - Genotypic patterns associated with nonnucleoside reverse transcriptase inhibitor (NNRTI) resistance in the absence of well-characterized resistance mutations were identified using a database (n > 47,000) of phenotype-genotype data. Among samples with no known NNRTI mutations, the most resistant samples contained K101P (n = 35) or a combination of K103R and V179D (n = 41). Site-directed mutagenesis confirmed the importance of these mutations. PMID- 16377710 TI - bla(IMP-9) and its association with large plasmids carried by Pseudomonas aeruginosa isolates from the People's Republic of China. AB - A novel plasmid-mediated metallo-beta-lactamase (IMP-9) is described in seven isolates of Pseudomonas aeruginosa from Guangzhou, China, isolated in 2000. The gene was carried on a large (approximately 450-kb) IncP-2 conjugative plasmid. This is the first report of carriage of bla(IMP) genes on such large plasmids. PMID- 16377711 TI - Heterogeneous macrolide resistance and gene conversion in the pneumococcus. AB - A macrolide-resistant clinical isolate of Streptococcus pneumoniae with 23S rRNA mutations showed a heterogeneous phenotype and genotype. The mutant 23S rRNA genes from this isolate transformed susceptible strain R6 to resistance. Culture of resistant strain R6 in the absence of antibiotic pressure showed gene conversion to occur between the four 23S rRNA alleles, resulting in reversion to susceptibility with the resistant phenotype showing a fitness cost. These data explain the disappearance on subculture of heterogeneous macrolide resistance in the pneumococcus. PMID- 16377712 TI - Lethal action of quinolones against a temperature-sensitive dnaB replication mutant of Escherichia coli. AB - Inhibition of DNA replication in an Escherichia coli dnaB-22 mutant failed to block quinolone-mediated lethality. Inhibition of protein synthesis by chloramphenicol inhibited nalidixic acid lethality and, to a lesser extent, ciprofloxacin lethality in both dnaB-22 and wild-type cells. Thus, major features of quinolone-mediated lethality do not depend on ongoing replication. PMID- 16377713 TI - Clonal dissemination and mupA gene polymorphism of mupirocin-resistant Staphylococcus aureus isolates from long-term-care facilities in South Korea. AB - We identified 25 high-level mupirocin-resistant (MuH) and 21 low-level mupirocin resistant (MuL) Staphylococcus aureus isolates from eight long-term-care facilities (LTCFs). The pulsed-field gel electrophoresis patterns of 19 MuH and 19 MuL isolates from two facilities were identical for 18 and 15 isolates, respectively. The most predominant mupA restriction fragment length polymorphism type was found in 21 MuH isolates. We conclude that clonal transmission of MuH and MuL S. aureus strains occurred in these LTCFs. This is the first report of clonal transfer of mupirocin resistance in LTCFs. PMID- 16377714 TI - In vitro activity of fosfomycin against extended-spectrum-beta-lactamase producing Escherichia coli and Klebsiella pneumoniae: comparison of susceptibility testing procedures. AB - The agar dilution, broth microdilution, and disk diffusion methods were compared to determine the in vitro susceptibility of 428 extended-spectrum-beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae to fosfomycin. Fosfomycin showed very high activity against all ESBL-producing strains. Excellent agreement between the three susceptibility methods was found for E. coli, whereas marked discrepancies were observed for K. pneumoniae. PMID- 16377715 TI - Norfloxacin blood-brain barrier transport in rats is not affected by probenecid coadministration. AB - The effect of probenecid (PRO) on norfloxacin (NOR) blood-brain barrier transport was investigated with rats by microdialysis. Maximum brain drug concentrations were rapidly attained, and the brain penetration factor was close to 5% in the absence and presence of PRO. In conclusion, PRO has no effect on NOR blood-brain barrier transport. PMID- 16377716 TI - Multicenter antimicrobial susceptibility survey of gram-negative bacteria isolated from patients with community-acquired infections in the People's Republic of China. AB - A survey of 2,099 gram-negative bacilli from community infections at seven centers in the People's Republic of China is reported. The rates of resistance of 1,615 isolates of the family Enterobacteriaceae were as follows: 40.8% for ciprofloxacin, 32.2% for gentamicin, 0% for imipenem or ertapenem, and 14.7% for cefotaxime. The rates of extended-spectrum beta-lactamase production were 16% for Escherichia coli and 17% for Klebsiella. PMID- 16377717 TI - Comparative in vitro activities of retapamulin (SB-275833) against 141 clinical isolates of Propionibacterium spp., including 117 P. acnes isolates. AB - Using the NCCLS agar dilution method, we studied the in vitro activity of retapamulin (SB-275833) against 141 clinical isolates of Propionibacterium species, including seven multiresistant strains, and found retapamulin to be the most active agent among those tested with MICs of < or = 1 microg/ml against all isolates. PMID- 16377718 TI - Arginine or nitrate enhances antibiotic susceptibility of Pseudomonas aeruginosa in biofilms. AB - Arginine enhanced the killing of Pseudomonas aeruginosa by ciprofloxacin and tobramycin under anaerobic, but not aerobic, growth conditions. Arginine or nitrate also enhanced the killing by these antibiotics in mature biofilms, reducing viable cell counts by a factor of 10 to 100 beyond that achieved by antibiotics alone. PMID- 16377719 TI - Different modes of action of naphthyridones in gram-positive and gram-negative bacteria. AB - Naphthyridones that were recently described as a class of translation inhibitors in gram-positive bacteria mediate their mode of action via GyrA in Haemophilus influenzae and Escherichia coli. These are the first examples of compounds in which modes of action in different bacterial pathogens are mediated through widely different targets. PMID- 16377720 TI - Bloodstream infections with metallo-beta-lactamase-producing Pseudomonas aeruginosa: epidemiology, microbiology, and clinical outcomes. AB - Pseudomonas aeruginosa strains that produce metallo-beta-lactamases (MBLs) are becoming increasingly prevalent. We evaluated the epidemiological and microbiological characteristics of monomicrobial bloodstream infections caused by MBL-producing P. aeruginosa isolates, as well as the clinical outcomes in patients with these infections. PMID- 16377721 TI - Dexamethasone attenuates staphylococcal enterotoxin B-induced hypothermic response and protects mice from superantigen-induced toxic shock. AB - The superantigenic staphylococcal enterotoxins are important virulence factors and contribute to various diseases, including food poisoning and toxic shock. Dexamethasone, an anti-inflammatory agent, attenuated staphylococcal enterotoxin B (SEB)-induced hypothermia and serum proinflammatory cytokines and improved survival from 0% to 86% in a lethal mouse model of SEB-mediated shock. PMID- 16377722 TI - Rifampin augments cytokine-induced nitric oxide production in human alveolar epithelial cells. AB - Rifampin increased nitric oxide production and inducible nitric oxide synthase expression in alveolar cells stimulated with cytokines. Nitric oxide concentrations after induction with cytokines, cytokines with 10 microg/ml rifampin, and cytokines with 50 microg/ml rifampin were 3.2, 4.5, and 8.8 microM, respectively (P < 0.02 versus cytokines alone). This indicates that rifampin modulates the immune response. PMID- 16377724 TI - Mutation in the Bacillus subtilis RNA polymerase beta' subunit confers resistance to lipiarmycin. PMID- 16377723 TI - OXA-58 and IMP-4 carbapenem-hydrolyzing beta-lactamases in an Acinetobacter junii blood culture isolate from Australia. PMID- 16377725 TI - Low correlation between MIC and mutant prevention concentration. PMID- 16377727 TI - Maximizing the use of scarce resources: vial optimization. PMID- 16377728 TI - IP-10/MCP-1 ratio in CSF is an useful diagnostic marker of neuropsychiatric lupus patients. PMID- 16377729 TI - Outcome of subacute low back pain: influence of patients' and rheumatologists' characteristics. AB - OBJECTIVES: To assess the outcome of subacute low back pain, to identify the characteristics of patients and physicians which were related to outcome and to evaluate the influence of rheumatologists' beliefs about back pain on their following the guidelines for physical and occupational activity. METHODS: DESIGN: a longitudinal descriptive survey. SETTING: a secondary care practice in France. PARTICIPANTS: 266 rheumatologists; 440 patients with subacute low back pain. OUTCOME MEASURES: the main outcome measure was persistence of back pain 3 months after baseline evaluation. A self-administered questionnaire for physicians assessed attitudes and beliefs about back pain [Fear-Avoidance Beliefs Questionnaire (FABQ)], and one for patients assessed pain, perceived handicap and disability (Quebec Scale), anxiety and depression (Hospital Anxiety Depression Questionnaire), and beliefs about back pain (FABQ). RESULTS: Forty per cent of patients had persistent low back pain at 3 months; 5.5% of these had sciatica. A total of 10% of rheumatologists and 68% of patients at baseline had a high FABQ physical score (phys; >14). Determinants of outcomes were work-related back pain [odds ratio (OR) = 3.37; 95% confidence interval (CI) 1.08-5.17], anxiety (OR = 2.41; 95% CI 1.44-4.09), sex (female OR = 2.03; 95% CI 1.30-3.18) and patients' beliefs about back pain at work (OR = 1.02; 95% CI 1.00-1.05). Physicians with high FABQ physical scores were less likely to follow guidelines on prescribing rest and occupational activity for back pain. CONCLUSION: Back pain commonly persists 3 months later in patients with subacute low back pain. Patients and rheumatologists still have negative beliefs about back pain. Rheumatologists' beliefs influence their following guidelines on physical and occupational activities. National education programmes about low back pain are needed in France. PMID- 16377730 TI - Two cases of serious food-borne infection in patients treated with anti-TNF alpha. Are we doing enough to reduce the risk? PMID- 16377731 TI - Disrupted cortical proprioceptive representation evokes symptoms of peculiarity, foreignness and swelling, but not pain. AB - OBJECTIVES: It has been proposed that disruption of the internal proprioceptive representation, via incongruent sensory input, may underpin pathological pain states, but experimental evidence relies on conflicting visual input, which is not clinically relevant. We aimed to determine the symptomatic effect of incongruent proprioceptive input, imparted by vibration of the wrist tendons, which evokes the illusion of perpetual wrist flexion and disrupts cortical proprioceptive representation. METHODS: Twenty-nine healthy and naive volunteers reported symptoms during five conditions: control, active and passive wrist flexion, extensor carpi radialis tendon vibration to evoke illusion of perpetual wrist flexion, and ulnar styloid (sham) vibration. No advice was given about possible illusions. RESULTS: Twenty-one subjects reported the illusion of perpetual wrist flexion during tendon vibration. There was no effect of condition or of whether or not subjects reported an illusion on discomfort/pain (P>0.28). Peculiarity, swelling and foreignness were greater during tendon vibration than during the other conditions, and greater during tendon vibration in those who reported an illusion of wrist flexion than in those who did not (P<0.05 for all). Symptoms were reported by at least two subjects in each condition and four subjects reported systemic symptoms (e.g. nausea). CONCLUSIONS: In healthy volunteers, incongruent proprioceptive input does not cause discomfort or pain but does evoke feelings of peculiarity, swelling and foreignness in the limb. PMID- 16377732 TI - Nitric oxide reduces seed dormancy in Arabidopsis. AB - Dormancy is a property of many mature seeds, and experimentation over the past century has identified numerous chemical treatments that will reduce seed dormancy. Nitrogen-containing compounds including nitrate, nitrite, and cyanide break seed dormancy in a range of species. Experiments are described here that were carried out to further our understanding of the mechanism whereby these and other compounds, such as the nitric oxide (NO) donor sodium nitroprusside (SNP), bring about a reduction in seed dormancy of Arabidopsis thaliana. A simple method was devised for applying the products of SNP photolysis through the gas phase. Using this approach it was shown that SNP, as well as potassium ferricyanide (Fe(III)CN) and potassium ferrocyanide (Fe(II)CN), reduced dormancy of Arabidopsis seeds by generating cyanide (CN). The effects of potassium cyanide (KCN) on dormant seeds were tested and it was confirmed that cyanide vapours were sufficient to break Arabidopsis seed dormancy. Nitrate and nitrite also reduced Arabidopsis seed dormancy and resulted in substantial rates of germination. The effects of CN, nitrite, and nitrate on dormancy were prevented by the NO scavenger c-PTIO. It was confirmed that NO plays a role in reducing seed dormancy by using purified NO gas, and a model to explain how nitrogen-containing compounds may break dormancy in Arabidopsis is presented. PMID- 16377733 TI - NO way to live; the various roles of nitric oxide in plant-pathogen interactions. AB - Nitric oxide has attracted considerable interest from plant pathologists due its established role in regulating mammalian anti-microbial defences, particularly via programmed cell death (PCD). Although NO plays a major role in plant PCD elicited in response to certain types of pathogenic challenge, the race-specific hypersensitive response (HR), it is now evident that NO also acts in the regulation of non-specific, papilla-based resistance to penetration by plant cells that survive attack and, possibly, in systemic acquired resistance. Equally, the potential roles of NO signalling/scavenging within the pathogen are being recognized. This review will consider key defensive roles played by NO in living cells during plant-pathogen interactions, as well as in those undergoing PCD. PMID- 16377734 TI - Modulation of nitric oxide bioactivity by plant haemoglobins. AB - Nitric oxide (NO) is a highly reactive signalling molecule that has numerous targets in plants. Both enzymatic and non-enzymatic synthesis of NO has been detected in several plant species, and NO functions have been characterized during diverse physiological processes such as plant growth, development, and resistance to biotic and abiotic stresses. This wide variety of effects reflects the basic signalling mechanisms that are utilized by virtually all mammalian and plant cells and suggests the necessity of detoxification mechanisms to control the level and functions of NO. During the last two years an increasing number of reports have implicated non-symbiotic haemoglobins as the key enzymatic system for NO scavenging in plants, indicating that the primordial function of haemoglobins may well be to protect against nitrosative stress and to modulate NO signalling functions. The biological relevance of plant haemoglobins during specific conditions of plant growth and stress, and the existence of further enzymatic and non-enzymatic NO scavenging systems, suggest the existence of precise NO modulation mechanisms in plants, as observed for different NO sources. PMID- 16377735 TI - Flower primordium formation at the Arabidopsis shoot apex: quantitative analysis of surface geometry and growth. AB - Geometry changes, especially surface expansion, accompanying flower primordium formation are investigated at the reproductive shoot apex of Arabidopsis with the aid of a non-invasive replica method and a 3-D reconstruction algorithm. The observed changes are characteristic enough to differentiate the early development of flower primordium in Arabidopsis into distinct stages. Primordium formation starts from the fast and anisotropic growth at the periphery of the shoot apical meristem, with the maximum extension in the meridional direction. Surprisingly, the primordium first becomes a shallow crease, and it is only later that this shape changes into a bulge. The bulge is formed from the shallow crease due to slower and less anisotropic growth than at the onset of primordium formation. It is proposed that the shallow crease is the first axil, i.e. the axil of a putative rudimentary bract subtending the flower primordium proper, while the flower primordium proper is the bulge formed at the bottom of this axil. At the adaxial side of the bulge, the second axil (a narrow and deep crease) is formed setting the boundary between the flower primordium proper and the shoot apical meristem. Surface growth, leading to the formation of the second axil, is slow and anisotropic. This is similar to the previously described growth pattern at the boundary of the leaf primordium in Anagallis. PMID- 16377736 TI - The two senescence-related markers, GS1 (cytosolic glutamine synthetase) and GDH (glutamate dehydrogenase), involved in nitrogen mobilization, are differentially regulated during pathogen attack and by stress hormones and reactive oxygen species in Nicotiana tabacum L. leaves. AB - To investigate the role of stress in nitrogen management in plants, the effect of pathogen attack, elicitors, and phytohormone application on the expression of the two senescence-related markers GS1 (cytosolic glutamine synthetase EC 6.3.1.2) and GDH (glutamate dehydrogenase, EC 1.4.1.2) involved in nitrogen mobilization in senescing leaves of tobacco (Nicotiana tabacum L.) plants, was studied. The expression of genes involved in primary nitrogen assimilation such as GS2 (chloroplastic glutamine synthetase) and Nia (nitrate reductase, EC 1.6.1.1) was also analysed. The Glubas gene, coding a beta-1,3-glucanase, was used as a plant defence gene control. As during natural senescence, the expression of GS2 and Nia was repressed under almost all stress conditions. By contrast, GS1 and GDH mRNA accumulation was increased. However, GS1 and GDH showed differential patterns of expression depending on the stress applied. The expression of GS1 appeared more selective than GDH. Results indicate that the GDH and GS1 genes involved in leaf senescence are also a component of the plant defence response during plant pathogen interaction. The links between natural plant senescence and stress induced senescence are discussed, as well as the potential role of GS1 and GDH in a metabolic safeguard process. PMID- 16377737 TI - The differential effects of herbivory by first and fourth instars of Trichoplusia ni (Lepidoptera: Noctuidae) on photosynthesis in Arabidopsis thaliana. AB - The effect of different feeding behaviours of 1st and 4th instar Trichoplusia ni on photosynthesis of Arabidopsis thaliana var. Columbia was characterized using spatially resolved measurements of fluorescence and leaf temperature, as well as leaf gas exchange,. First instars made small holes with a large perimeter-to-area ratio and avoided veins, while 4th instars made large holes with a low perimeter to-area ratio and consumed veins. Herbivory by 1st instars reduced photosynthesis more strongly in the remaining leaf tissue than that by 4th instars. Photosystem II operating efficiency (PhiPSII) was correlated with the rate of CO2 exchange, and reductions in PhiPSII in areas around the missing tissues contributed to a 15.6% reduction in CO2 assimilation on the first day following removal of 1st instars. The corresponding increases in non-photochemical quenching and greater rates of non-stomatal water loss from these regions, as well as the partial reversal of low PhiPSII by increasing the ambient CO2 concentration, suggests that localized water stress and reduced stomatal conductance contributed to the inhibition of photosynthesis. Damage by 1st but not 4th instars reduced the maximum quantum efficiency of photosystem II photochemistry (Fv/Fm) by 4-8%. While herbivory by both 1st and 4th instars increased dark respiration rates, the rates were too low to have contributed to the observed reductions in CO2 exchange. The small holes produced by 1st instars may have isolated patches of tissue from the vascular system thereby contributing to localized water stress. Since neither 1st nor 4th instar herbivory had a detectable effect on the expression of the Rubisco small subunit gene, the observed differences cannot be attributed to changes in expression of this gene. The mode of feeding by different instars of T. ni determined the photosynthetic response to herbivory, which appeared to be mediated by the level of water stress associated with herbivore damage. PMID- 16377739 TI - Comparison of breast cancer mortality in five countries: France, Italy, Japan, the UK and the USA from the WHO mortality database (1960-2000). PMID- 16377740 TI - The onset of gravisensitivity in the embryonic root of flax. AB - Vertical orientation of emerging roots typically is the first response of plants to gravity. Although root gravitropism has been studied extensively, no conclusive data on the onset of gravisensing exist. We determined the inception of gravisensitivity in flax (Linum usitatissimum) roots by clinorotating germinating seeds after various periods of static orientation (gravistimulation) of imbibed seeds. Gravitropic competency was established about 8 h after imbibition, 11 h prior to germination. The time was determined based on 50% of the newly emerged roots curving in the direction of the gravity vector during static imbibition, despite subsequent clinorotation. The threshold value was affected by the orientation of the seeds. Upward orientation of the micropyle/radicle reduced the number of graviresponding roots to about one-half. Prolonged clinorotation weakened the graviresponse. Gravisensing was accompanied by the development of amyloplasts, but the actin cytoskeleton was not involved because imbibition in Latrunculin B did not affect the onset of gravisensitivity or germination, and the development of F-actin in untreated controls was observed only after the onset of gravisensitivity. PMID- 16377741 TI - Identification of the flavonoid hydroxylases from grapevine and their regulation during fruit development. AB - Flavonoids are important secondary metabolites in many fruits, and their hydroxylation pattern determines their color, stability, and antioxidant capacity. Hydroxylation of the B-ring of flavonoids is catalyzed by flavonoid 3' hydroxylase (F3'H) and flavonoid 3',5'-hydroxylase (F3'5'H), and may also require cytochrome b5. We report the identification of genes encoding F3'H, F3'5'H, and a putative cytochrome b5 from grapevine (Vitis vinifera L. cv Shiraz) and their transcriptional regulation in fruit. Functionality of the genes VvF3'H and VvF3'5'H1 was demonstrated by ectopic expression in petunia (Petunia hybrida), which altered flower color and flavonoid composition as expected. VvF3'H was expressed in grapes before flowering, when 3'-hydroxylated flavonols are made, and all three genes were expressed after flowering, when proanthocyanidins (PAs) are synthesized. In berry skin, expression of all three genes was low at the onset of ripening (veraison) but increased after veraison concomitant with the accumulation of 3'- and 3',5'-hydroxylated anthocyanins. VvF3'H and VvCytoB5 were expressed in seeds but not VvF3'5'H1, consistent with the accumulation of 3' hydroxylated PAs in this tissue. VvCytoB5 expression was correlated with expression of both VvF3'H and VvF3'5'H1 in the different grape tissues. In contrast to red grapes, where VvF3'H, VvF3'5'H1, and VvCytoB5 were highly expressed during ripening, the expression of VvF3'5'H1 and VvCytoB5 in white grapes during ripening was extremely low, suggesting a difference in transcriptional regulation. Our results show that temporal and tissue-specific expression of VvF3'H, VvF3'5'H1, and VvCytoB5 in grapes is coordinated with the accumulation of the respective hydroxylated flavonols and PAs, as well as anthocyanins. Understanding the regulation of flavonoid hydroxylases could be used to modify flavonoid composition of fruits. PMID- 16377742 TI - Pea seed mitochondria are endowed with a remarkable tolerance to extreme physiological temperatures. AB - Most seeds are anhydrobiotes, relying on an array of protective and repair mechanisms, and seed mitochondria have previously been shown to harbor stress proteins probably involved in desiccation tolerance. Since temperature stress is a major issue for germinating seeds, the temperature response of pea (Pisum sativum) seed mitochondria was examined in comparison with that of mitochondria from etiolated epicotyl, a desiccation-sensitive tissue. The functional analysis illustrated the remarkable temperature tolerance of seed mitochondria in response to both cold and heat stress. The mitochondria maintained a well-coupled respiration between -3.5 degrees C and 40 degrees C, while epicotyl mitochondria were not efficient below 0 degrees C and collapsed above 30 degrees C. Both mitochondria exhibited a similar Arrhenius break temperature at 7 degrees C, although they differed in phospholipid composition. Seed mitochondria had a lower phosphatidylethanolamine-to-phosphatidylcholine ratio, fewer unsaturated fatty acids, and appeared less susceptible to lipid peroxidation. They also accumulated large amounts of heat shock protein HSP22 and late-embryogenesis abundant protein PsLEAm. The combination of membrane composition and stress protein accumulation required for desiccation tolerance is expected to lead to an unusually wide temperature tolerance, contributing to the fitness of germinating seeds in adverse conditions. The unique oxidation of external NADH at low temperatures found with several types of mitochondria may play a central role in maintaining energy homeostasis during cold shock, a situation often encountered by sessile and ectothermic higher plants. PMID- 16377743 TI - ARABINAN DEFICIENT 1 is a putative arabinosyltransferase involved in biosynthesis of pectic arabinan in Arabidopsis. AB - The function of a putative glycosyltransferase (At2g35100) was investigated in Arabidopsis (Arabidopsis thaliana). The protein is predicted to be a type 2 membrane protein with a signal anchor. Two independent mutant lines with T-DNA insertion in the ARABINAN DEFICIENT 1 (ARAD1) gene were analyzed. The gene was shown to be expressed in all tissues but particularly in vascular tissues of leaves and stems. Analysis of cell wall polysaccharides isolated from leaves and stems showed that arabinose content was reduced to about 75% and 46%, respectively, of wild-type levels. Immunohistochemical analysis indicated a specific decrease in arabinan with no change in other pectic domains or in glycoproteins. The cellular structure of the stem was also not altered. Isolated rhamnogalacturonan I from mutant tissues contained only about 30% of the wild type amount of arabinose, confirming the specific deficiency in arabinan. Linkage analysis showed that the small amount of arabinan present in mutant tissue was structurally similar to that of the wild type. Transformation of mutant plants with the ARAD1 gene driven by the 35S promoter led to full complementation of the phenotype, but none of the transformants had more arabinan than the wild-type level. The data suggest that ARAD1 is an arabinan alpha-1,5 arabinosyltransferase. To our knowledge, the identification of other L arabinosyltransferases has not been published. PMID- 16377744 TI - The outcomes of concentration-specific interactions between salicylate and jasmonate signaling include synergy, antagonism, and oxidative stress leading to cell death. AB - Salicylic acid (SA) has been proposed to antagonize jasmonic acid (JA) biosynthesis and signaling. We report, however, that in salicylate hydroxylase expressing tobacco (Nicotiana tabacum) plants, where SA levels were reduced, JA levels were not elevated during a hypersensitive response elicited by Pseudomonas syringae pv phaseolicola. The effects of cotreatment with various concentrations of SA and JA were assessed in tobacco and Arabidopsis (Arabidopsis thaliana). These suggested that there was a transient synergistic enhancement in the expression of genes associated with either JA (PDF1.2 [defensin] and Thi1.2 [thionin]) or SA (PR1 [PR1a-beta-glucuronidase in tobacco]) signaling when both signals were applied at low (typically 10-100 microm) concentrations. Antagonism was observed at more prolonged treatment times or at higher concentrations. Similar results were also observed when adding the JA precursor, alpha-linolenic acid with SA. Synergic effects on gene expression and plant stress were NPR1- and COI1-dependent, SA- and JA-signaling components, respectively. Electrolyte leakage and Evans blue staining indicated that application of higher concentrations of SA + JA induced plant stress or death and elicited the generation of apoplastic reactive oxygen species. This was indicated by enhancement of hydrogen peroxide-responsive AoPR10-beta-glucuronidase expression, suppression of plant stress/death using catalase, and direct hydrogen peroxide measurements. Our data suggests that the outcomes of JA-SA interactions could be tailored to pathogen/pest attack by the relative concentration of each hormone. PMID- 16377745 TI - Transcript analysis of early nodulation events in Medicago truncatula. AB - Within the first 72 h of the interaction between rhizobia and their host plants, nodule primordium induction and infection occur. We predicted that transcription profiling of early stages of the symbiosis between Medicago truncatula roots and Sinorhizobium meliloti would identify regulated plant genes that likely condition key events in nodule initiation. Therefore, using a microarray with about 6,000 cDNAs, we compared transcripts from inoculated and uninoculated roots corresponding to defined stages between 1 and 72 h post inoculation (hpi). Hundreds of genes of both known and unknown function were significantly regulated at these time points. Four stages of the interaction were recognized based on gene expression profiles, and potential marker genes for these stages were identified. Some genes that were regulated differentially during stages I (1 hpi) and II (6-12 hpi) of the interaction belong to families encoding proteins involved in calcium transport and binding, reactive oxygen metabolism, and cytoskeleton and cell wall functions. Genes involved in cell proliferation were found to be up-regulated during stages III (24-48 hpi) and IV (72 hpi). Many genes that are homologs of defense response genes were up-regulated during stage I but down-regulated later, likely facilitating infection thread progression into the root cortex. Additionally, genes putatively involved in signal transduction and transcriptional regulation were found to be differentially regulated in the inoculated roots at each time point. The findings shed light on the complexity of coordinated gene regulation and will be useful for continued dissection of the early steps in symbiosis. PMID- 16377746 TI - Cell wall proteome in the maize primary root elongation zone. I. Extraction and identification of water-soluble and lightly ionically bound proteins. AB - Cell wall proteins (CWPs) play important roles in various processes, including cell elongation. However, relatively little is known about the composition of CWPs in growing regions. We are using a proteomics approach to gain a comprehensive understanding of the identity of CWPs in the maize (Zea mays) primary root elongation zone. As the first step, we examined the effectiveness of a vacuum infiltration-centrifugation technique for extracting water-soluble and loosely ionically bound (fraction 1) CWPs from the root elongation zone. The purity of the CWP extract was evaluated by comparing with total soluble proteins extracted from homogenized tissue. Several lines of evidence indicated that the vacuum infiltration-centrifugation technique effectively enriched for CWPs. Protein identification revealed that 84% of the CWPs were different from the total soluble proteins. About 40% of the fraction 1 CWPs had traditional signal peptides and 33% were predicted to be nonclassical secretory proteins, whereas only 3% and 11%, respectively, of the total soluble proteins were in these categories. Many of the CWPs have previously been shown to be involved in cell wall metabolism and cell elongation. In addition, maize has type II cell walls, and several of the CWPs identified in this study have not been identified in previous cell wall proteomics studies that have focused only on type I walls. These proteins include endo-1,3;1,4-beta-D-glucanase and alpha-L arabinofuranosidase, which act on the major polysaccharides only or mainly present in type II cell walls. PMID- 16377747 TI - MICROTUBULE ORGANIZATION 1 regulates structure and function of microtubule arrays during mitosis and cytokinesis in the Arabidopsis root. AB - MICROTUBULE ORGANIZATION 1 (MOR1) is a plant member of the highly conserved MAP215/Dis1 family of microtubule-associated proteins. Prior studies with the temperature-sensitive mor1 mutants of Arabidopsis (Arabidopsis thaliana), which harbor single amino acid substitutions in an N-terminal HEAT repeat, proved that MOR1 regulates cortical microtubule organization and function. Here we demonstrate by use of live cell imaging and immunolabeling that the mor1-1 mutation generates specific defects in the microtubule arrays of dividing vegetative cells. Unlike the universal cortical microtubule disorganization in elongating mor1-1 cells, disruption of mitotic and cytokinetic microtubule arrays was not detected in all dividing cells. Nevertheless, quantitative analysis identified distinct defects in preprophase bands (PPBs), spindles, and phragmoplasts. In nearly one-half of dividing cells at the restrictive temperature of 30 degrees C, PPBs were not detected prior to spindle formation, and those that did form were often disrupted. mor1-1 spindles and phragmoplasts were short and abnormally organized and persisted for longer times than in wild type cells. The reduced length of these arrays predicts that the component microtubule lengths are also reduced, suggesting that microtubule length is a critical determinant of spindle and phragmoplast structure, orientation, and function. Microtubule organizational defects led to aberrant chromosomal arrangements, misaligned or incomplete cell plates, and multinucleate cells. Antiserum raised against an N-terminal MOR1 sequence labeled the full length of microtubules in interphase arrays, PPBs, spindles, and phragmoplasts. Continued immunolabeling of the disorganized and short microtubules of mor1-1 at the restrictive temperature demonstrated that the mutant mor1-1(L174F) protein loses function without dissociating from microtubules, providing important insight into the mechanism by which MOR1 may regulate microtubule length. PMID- 16377749 TI - The DMI1 and DMI2 early symbiotic genes of medicago truncatula are required for a high-affinity nodulation factor-binding site associated to a particulate fraction of roots. AB - The establishment of the legume-rhizobia symbiosis between Medicago spp. and Sinorhizobium meliloti is dependent on the production of sulfated lipo chitooligosaccharidic nodulation (Nod) factors by the bacterial partner. In this article, using a biochemical approach to characterize putative Nod factor receptors in the plant host, we describe a high-affinity binding site (Kd = 0.45 nm) for the major Nod factor produced by S. meliloti. This site is termed Nod factor-binding site 3 (NFBS3). NFBS3 is associated to a high-density fraction prepared from roots of Medicago truncatula and shows binding specificity for lipo chitooligosaccharidic structures. As for the previously characterized binding sites (NFBS1 and NFBS2), NFBS3 does not recognize the sulfate group on the S. meliloti Nod factor. Studies of Nod factor binding in root extracts of early symbiotic mutants of M. truncatula reveals that the new site is present in Nod factor perception and does not make infections 3 (dmi3) mutants but is absent in dmi1 and dmi2 mutants. Roots and cell cultures of all these mutants still contain sites similar to NFBS1 and NFBS2, respectively. These results suggest that NFBS3 is different from NFBS2 and NFBS1 and is dependent on the common symbiotic genes DMI1 and DMI2 required for establishment of symbioses with both rhizobia and arbuscular mycorrhizal fungi. The potential role of this site in the establishment of root endosymbioses is discussed. PMID- 16377748 TI - A coumaroyl-ester-3-hydroxylase insertion mutant reveals the existence of nonredundant meta-hydroxylation pathways and essential roles for phenolic precursors in cell expansion and plant growth. AB - Cytochromes P450 monooxygenases from the CYP98 family catalyze the meta hydroxylation step in the phenylpropanoid biosynthetic pathway. The ref8 Arabidopsis (Arabidopsis thaliana) mutant, with a point mutation in the CYP98A3 gene, was previously described to show developmental defects, changes in lignin composition, and lack of soluble sinapoyl esters. We isolated a T-DNA insertion mutant in CYP98A3 and show that this mutation leads to a more drastic inhibition of plant development and inhibition of cell growth. Similar to the ref8 mutant, the insertion mutant has reduced lignin content, with stem lignin essentially made of p-hydroxyphenyl units and trace amounts of guaiacyl and syringyl units. However, its roots display an ectopic lignification and a substantial proportion of guaiacyl and syringyl units, suggesting the occurrence of an alternative CYP98A3-independent meta-hydroxylation mechanism active mainly in the roots. Relative to the control, mutant plantlets produce very low amounts of sinapoyl esters, but accumulate flavonol glycosides. Reduced cell growth seems correlated with alterations in the abundance of cell wall polysaccharides, in particular decrease in crystalline cellulose, and profound modifications in gene expression and homeostasis reminiscent of a stress response. CYP98A3 thus constitutes a critical bottleneck in the phenylpropanoid pathway and in the synthesis of compounds controlling plant development. CYP98A3 cosuppressed lines show a gradation of developmental defects and changes in lignin content (40% reduction) and structure (prominent frequency of p-hydroxyphenyl units), but content in foliar sinapoyl esters is similar to the control. The purple coloration of their leaves is correlated to the accumulation of sinapoylated anthocyanins. PMID- 16377750 TI - Phloem loading in two Scrophulariaceae species. What can drive symplastic flow via plasmodesmata? AB - To determine the driving forces for symplastic sugar flux between mesophyll and phloem, gradients of sugar concentrations and osmotic pressure were studied in leaf tissues of two Scrophulariaceae species, Alonsoa meridionalis and Asarina barclaiana. A. meridionalis has a typical symplastic configuration of minor-vein phloem, i.e. intermediary companion cells with highly developed plasmodesmal connections to bundle-sheath cells. In A. barclaiana, two types of companion cells, modified intermediary cells and transfer cells, were found in minor-vein phloem, giving this species the potential to have a complex phloem-loading mode. We identified all phloem-transported carbohydrates in both species and analyzed the levels of carbohydrates in chloroplasts, vacuoles, and cytoplasm of mesophyll cells by nonaqueous fractionation. Osmotic pressure was measured in single epidermal and mesophyll cells and in whole leaves and compared with calculated values for phloem sap. In A. meridionalis, a 2-fold concentration gradient for sucrose between mesophyll and phloem was found. In A. barclaiana, the major transported carbohydrates, sucrose and antirrhinoside, were present in the phloem in 22- and 6-fold higher concentrations, respectively, than in the cytoplasm of mesophyll cells. The data show that diffusion of sugars along their concentration gradients is unlikely to be the major mechanism for symplastic phloem loading if this were to occur in these species. We conclude that in both A. meridionalis and A. barclaiana, apoplastic phloem loading is an indispensable mechanism and that symplastic entrance of solutes into the phloem may occur by mass flow. The conditions favoring symplastic mass flow into the phloem are discussed. PMID- 16377751 TI - Analysis of freeze-thaw embolism in conifers. The interaction between cavitation pressure and tracheid size. AB - Ice formation in the xylem sap produces air bubbles that under negative xylem pressures may expand and cause embolism in the xylem conduits. We used the centrifuge method to evaluate the relationship between freeze-thaw embolism and conduit diameter across a range of xylem pressures (Px) in the conifers Pinus contorta and Juniperus scopulorum. Vulnerability curves showing loss of conductivity (embolism) with Px down to -8 MPa were generated with versus without superimposing a freeze-thaw treatment. In both species, the freeze-thaw plus water-stress treatment caused more embolism than water stress alone. We estimated the critical conduit diameter (Df) above which a tracheid will embolize due to freezing and thawing and found that it decreased from 35 microm at a Px of -0.5 MPa to 6 microm at -8 MPa. Further analysis showed that the proportionality between diameter of the air bubble nucleating the cavitation and the diameter of the conduit (kL) declined with increasingly negative Px. This suggests that the bubbles causing cavitation are smaller in proportion to tracheid diameter in narrow tracheids than in wider ones. A possible reason for this is that the rate of dissolving increases with bubble pressure, which is inversely proportional to bubble diameter (La Place's law). Hence, smaller bubbles shrink faster than bigger ones. Last, we used the empirical relationship between Px and Df to model the freeze-thaw response in conifer species. PMID- 16377752 TI - Proteomic analysis of different mutant genotypes of Arabidopsis led to the identification of 11 proteins correlating with adventitious root development. AB - A lack of competence to form adventitious roots by cuttings or explants in vitro occurs routinely and is an obstacle for the clonal propagation and rapid fixation of elite genotypes. Adventitious rooting is known to be a quantitative genetic trait. We performed a proteomic analysis of Arabidopsis (Arabidopsis thaliana) mutants affected in their ability to develop adventitious roots in order to identify associated molecular markers that could be used to select genotypes for their rooting ability and/or to get further insight into the molecular mechanisms controlling adventitious rooting. Comparison of two-dimensional gel electrophoresis protein profiles resulted in the identification of 11 proteins whose abundance could be either positively or negatively correlated with endogenous auxin content, the number of adventitious root primordia, and/or the number of mature adventitious roots. One protein was negatively correlated only to the number of root primordia and two were negatively correlated to the number of mature adventitious roots. Two putative chaperone proteins were positively correlated only to the number of primordia, and, interestingly, three auxin inducible GH3-like proteins were positively correlated with the number of mature adventitious roots. The others were correlated with more than one parameter. The 11 proteins are predicted to be involved in different biological processes, including the regulation of auxin homeostasis and light-associated metabolic pathways. The results identify regulatory pathways associated with adventitious root formation and represent valuable markers that might be used for the future identification of genotypes with better rooting abilities. PMID- 16377753 TI - Patterns of sequence loss and cytosine methylation within a population of newly resynthesized Brassica napus allopolyploids. AB - Allopolyploid formation requires the adaptation of two nuclear genomes within a single cytoplasm, which may involve programmed genetic and epigenetic changes during the initial generations following genome fusion. To study the dynamics of genome change, we synthesized 49 isogenic Brassica napus allopolyploids and surveyed them with 76 restriction fragment length polymorphism (RFLP) probes and 30 simple sequence repeat (SSR) primer pairs. Here, we report on the types and distribution of genetic and epigenetic changes within the S(1) genotypes. We found that insertion/deletion (indel) events were rare, but not random. Of the 57,710 (54,383 RFLP and 3,327 SSR) parental fragments expected among the amphidiploids, we observed 56,676 or 99.9%. Three loci derived from Brassica rapa had indels, and one indel occurred repeatedly across 29% (14/49) of the lines. Loss of one parental fragment was due to the 400-bp reduction of a guanine adenine dinucleotide repeat-rich sequence. In contrast to the 4% (3/76) RFLP probes that detected indels, 48% (35/73) detected changes in the CpG methylation status between parental genomes and the S1 lines. Some loci were far more likely than others to undergo epigenetic change, but the number of methylation changes within each synthetic polyploid was remarkably similar to others. Clear de novo methylation occurred at a much higher frequency than de novo demethylation within allopolyploid sequences derived from B. rapa. Our results suggest that there is little genetic change in the S(0) generation of resynthesized B. napus polyploids. In contrast, DNA methylation was altered extensively in a pattern that indicates tight regulation of epigenetic changes. PMID- 16377754 TI - A defect in exodegradative pathways provides insight into endodegradation of heparan and dermatan sulfates. AB - Within cells, dermatan sulfate (DS) and heparan sulfate (HS) are degraded in two steps. The initial endohydrolysis of these polysaccharides is followed by the sequential action of lysosomal exoenzymes to reduce the resulting oligosaccharides to monosaccharides and inorganic sulfate. Mucopolysaccharidosis (MPS) type II is a lysosomal storage disorder caused by a deficiency of the exoenzyme iduronate-2-sulfatase (I2S). Consequently, partially degraded fragments of DS and HS have been shown to accumulate in the lysosomes of affected cells and are excreted in the urine. Di- to hexadecasaccharides, isolated from the urine of a MPS II patient using anion exchange and gel filtration chromatography, were identified using electrospray ionization-tandem mass spectrometry (ESI-MS/MS). These oligosaccharides were shown to have non-reducing terminal iduronate-2 sulfate residues by digestion with recombinant I2S. A pattern of growing oligosaccharide chains composed of alternating uronic acid and N-acetylhexosamine residues was identified and suggested to originate from DS. A series of oligosaccharides consisting of hexosamine/N-acetylhexosamine alternating with uronic acid residues was also identified and on the basis of the presence of unacetylated hexosamine; these oligosaccharides are proposed to derive from HS. The presence of both odd and even-length oligosaccharides suggests both endo-beta glucuronidase and endo-N-acetylhexosaminidase activities toward both glycosaminoglycans. Furthermore, the putative HS oligosaccharide structures identified indicate that heparanase activities are directed toward regions of both low and high sulfation, while the N-acetylhexosaminidase activity acted only in regions of low sulfation in this polysaccharide. PMID- 16377755 TI - William Pulteney Alison, the Scottish philosophy, and the making of a political medicine. AB - This article considers the public health and social-reform agitations of Dr. William Pulteney Alison (1790-1858), professor of medicine at Edinburgh University and leader of the Scottish medical profession, in the context of Scottish moral philosophy. Throughout his career, Alison reflected on what has come to be recognized as a central problem of social medicine: where did its domain end? At what point did the medical mission of identifying and eliminating factors that harm health pass into a non-medical domain-the provinces of political economy, individual liberty, participatory politics, or acceptance of nature's dictates? On these issues Alison was an expansionist, relentlessly pushing back the borders of medicine. Drawing on Alison's writings on such disparate topics as the philosophy of mind, the epidemiology of infectious diseases, and modes of agrarian organization, the article argues that the trajectory of much of Alison's work was to discover the structural implications of a comprehensive biological reading of human capacity and behavior. It is therefore appropriate to see him as a promulgator of a "political medicine," which he presented as a critical alternative to the classical political economy of the Scottish Malthusians. The article concludes by suggesting that Alison's work (and influence) have been under-recognized and remain pertinent to modern social epidemiology, public health, and medicine more broadly. PMID- 16377756 TI - Increased expression of MAP KINASE KINASE7 causes deficiency in polar auxin transport and leads to plant architectural abnormality in Arabidopsis. AB - Polar auxin transport (PAT) plays a crucial role in the regulation of many aspects of plant growth and development. We report the characterization of a semidominant Arabidopsis thaliana bushy and dwarf1 (bud1) mutant. Molecular genetic analysis indicated that the bud1 phenotype is a result of increased expression of Arabidopsis MAP KINASE KINASE7 (MKK7), a member of plant mitogen activated protein kinase kinase group D. We showed that BUD1/MKK7 is a functional kinase and that the kinase activity is essential for its biological functions. Compared with the wild type, the bud1 plants develop significantly fewer lateral roots, simpler venation patterns, and a quicker and greater curvature in the gravitropism assay. In addition, the bud1 plants have shorter hypocotyls at high temperature (29 degrees C) under light, which is a characteristic feature of defective auxin action. Determination of tritium-labeled indole-3-acetic acid transport showed that the increased expression of MKK7 in bud1 or the repressed expression in MKK7 antisense transgenic plants causes deficiency or enhancement in auxin transport, indicating that MKK7 negatively regulates PAT. This conclusion was further substantiated by genetic and phenotypic analyses of double mutants generated from crosses between bud1 and the auxin-related mutants axr3-3, tir1-1, doc1-1, and atmdr1-1. PMID- 16377757 TI - A rice glutamate receptor-like gene is critical for the division and survival of individual cells in the root apical meristem. AB - Glu receptors are known to function as Glu-activated ion channels that mediate mostly excitatory neurotransmission in animals. Glu receptor-like genes have also been reported in higher plants, although their function is largely unknown. We have identified a rice (Oryza sativa) Glu receptor-like gene, designated GLR3.1, in which mutation by T-DNA insertion caused a short-root mutant phenotype. Histology and DNA synthesis analyses revealed that the mutant root meristematic activity is distorted and is accompanied by enhanced programmed cell death. Our results supply genetic evidence that a plant Glu receptor-like gene, rice GLR3.1, is essential for the maintenance of cell division and individual cell survival in the root apical meristem at the early seedling stage. PMID- 16377758 TI - The Arabidopsis receptor kinase FLS2 binds flg22 and determines the specificity of flagellin perception. AB - Flagellin, the main building block of the bacterial flagellum, acts as a pathogen associated molecular pattern triggering the innate immune response in animals and plants. In Arabidopsis thaliana, the Leu-rich repeat transmembrane receptor kinase FLAGELLIN SENSITIVE2 (FLS2) is essential for flagellin perception. Here, we demonstrate the specific interaction of the elicitor-active epitope flg22 with the FLS2 protein by chemical cross-linking and immunoprecipitation. The functionality of this receptor was further tested by heterologous expression of the Arabidopsis FLS2 gene in tomato (Lycopersicon esculentum) cells. The perception of flg22 in tomato differs characteristically from that in Arabidopsis. Expression of Arabidopsis FLS2 conferred an additional flg22 perception system on the cells of tomato, which showed all of the properties characteristic of the perception of this elicitor in Arabidopsis. In summary, these results show that FLS2 constitutes the pattern-recognition receptor that determines the specificity of flagellin perception. PMID- 16377760 TI - Microarray analysis of mouse ear tissue exposed to bis-(2-chloroethyl) sulfide: gene expression profiles correlate with treatment efficacy and an established clinical endpoint. AB - Bis-(2-chloroethyl) sulfide (sulfur mustard; SM) is a potent alkylating agent. Three treatment compounds have been shown to limit SM damage in the mouse ear vesicant model: dimercaprol, octyl homovanillamide, and indomethacin. Microarrays were used to determine gene expression profiles of biopsies taken from mouse ears after exposure to SM in the presence or absence of treatment compounds. Mouse ears were topically exposed to SM alone or were pretreated for 15 min with a treatment compound and then exposed to SM. Ear tissue was harvested 24 h after exposure for ear weight determination, the endpoint used to evaluate treatment compound efficacy. RNA extracted from the tissues was used to generate microarray probes for gene expression profiling of therapeutic responses. Principal component analysis of the gene expression data revealed partitioning of the samples based on treatment compound and SM exposure. Patterns of gene responses to the treatment compounds were indicative of exposure condition and were phenotypically anchored to ear weight. Pretreatment with indomethacin, the least effective treatment compound, produced ear weights close to those treated with SM alone. Ear weights from animals pretreated with dimercaprol or octyl homovanillamide were more closely associated with exposure to vehicle alone. Correlation coefficients between gene expression level and ear weight revealed genes involved in mediating responses to both SM exposure and treatment compounds. These data provide a basis for elucidating the mechanisms of response to SM and drug treatment and also provide a basis for developing strategies to accelerate development of effective SM medical countermeasures. PMID- 16377759 TI - Arabidopsis TARGET OF RAPAMYCIN interacts with RAPTOR, which regulates the activity of S6 kinase in response to osmotic stress signals. AB - TARGET OF RAPAMYCIN (TOR) kinase controls many cellular functions in eukaryotic cells in response to stress and nutrient availability and was shown to be essential for embryonic development in Arabidopsis thaliana. We demonstrated that Arabidopsis RAPTOR1 (a TOR regulatory protein) interacts with the HEAT repeats of TOR and that RAPTOR1 regulates the activity of S6 kinase (S6K) in response to osmotic stress. RAPTOR1 also interacts in vivo with Arabidopsis S6K1, a putative substrate for TOR. S6K1 fused to green fluorescent protein and immunoprecipitated from tobacco (Nicotiana tabacum) leaves after transient expression was active in phosphorylating the Arabidopsis ribosomal S6 protein. The catalytic domain of S6K1 could be phosphorylated by Arabidopsis 3-phosphoinositide-dependent protein kinase-1 (PDK1), indicating the involvement of PDK1 in the regulation of S6K. The S6K1 activity was sensitive to osmotic stress, while PDK1 activity was not affected. However, S6K1 sensitivity to osmotic stress was relieved by co overexpression of RAPTOR1. Overall, these observations demonstrated the existence of a functional TOR kinase pathway in plants. However, Arabidopsis seedlings do not respond to normal physiological levels of rapamycin, which appears to be due its inability to bind to the Arabidopsis homolog of FKBP12, a protein that is essential for the binding of rapamycin with TOR. Replacement of the Arabidopsis FKBP12 with the human FKBP12 allowed rapamycin-dependent interaction with TOR. Since homozygous mutation in TOR is lethal, it suggests that this pathway is essential for integrating the stress signals into the growth regulation. PMID- 16377761 TI - Alteration in erythropoietin-induced cardioprotective signaling by postinfarct ventricular remodeling. AB - Postinfarct remodeling impairs mechanisms of ischemic preconditioning. We examined whether myocardial response to activation of the erythropoietin (EPO) receptor is modified by postinfarct remodeling. Four weeks after induction of myocardial infarction (MI) by coronary ligation in post-MI group (post-MI) or a sham operation in sham group (sham), rat hearts were isolated and subjected to 25 min global ischemia/2-h reperfusion. Infarct size was expressed as a percentage of risk area (i.e., left ventricle) from which scarred infarct was excluded (%I/R). The heart weight was 15% larger in post-MI, but there was no intergroup difference in plasma EPO levels or myocardial EPO receptor levels. EPO infusion (5 U/ml) significantly reduced %I/R from 59.9 +/- 4.1 to 36.2 +/- 4.2 in sham and from 58.1 +/- 5.0 to 35.2 +/- 4.0 in post-MI. This EPO-induced protection was sensitive to a phosphatidylinositol 3-kinase (PI3K) inhibitor, 2-(4-morpholinyl) 8-phenyl-4H-1-benzopyran-4-one (LY294002), in sham. However, neither LY294002 nor wortmannin inhibited the EPO-induced protection in post-MI. Phosphorylation of Janus kinase 2 by EPO was attenuated and phosphorylation of Akt was not detected in post-MI. A guanylyl cyclase inhibitor, 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1 one, and a mitochondrial ATP-sensitive K(+) channel (mitoK(ATP) channel) blocker, 5-hydroxydecanoate, inhibited EPO-induced protection in both sham and post-MI. Suppressor of cytokine signaling (SOCS)-1 protein level was higher by 50% in post MI than in sham, although SOCS-3 levels were similar. These findings suggest that postinfarct remodeling disrupts cellular signaling from the EPO receptor to PI3K, presumably by increased SOCS-1. However, in the remodeled myocardium, lack of PI3K/Akt activation by the EPO receptor seems to be compensated by a mechanism upstream of the guanylyl cyclase-mitoK(ATP) channel pathway to achieve EPO induced protection. PMID- 16377762 TI - Endothelial nitric-oxide synthase reveals a new face in G protein signaling. AB - In this issue of Molecular Pharmacology, Andreeva et al. (p. 975) report a novel functional link between the heterotrimeric G protein G alpha12 and endothelial nitric-oxide synthase (eNOS). Based on studies characterizing the interaction of G alpha12 and the molecular chaperone Hsp90 and the interaction of eNOS and Hsp90, the group proposed an interaction between G alpha12 and eNOS and sought to determine the regulatory mechanisms, including the inferred dependence on Hsp90. Their experiments using an overexpression model lead to the observation that the cotransfection of G alpha12 and eNOS expression vectors increased overall eNOS expression. Additional studies in the overexpression model and in human umbilical vein endothelial cells (HUVEC) provide evidence for a mechanism that involves G alpha12-dependent stabilization of eNOS protein and possibly mRNA. These data present yet another paradigm by which heterotrimeric G proteins, through stabilization of target proteins, can regulate the activity of downstream signaling pathways. PMID- 16377764 TI - Serine 202 regulates the nuclear translocation of constitutive active/androstane receptor. AB - The constitutive active receptor (CAR) in mouse primary hepatocytes undergoes okadaic acid (OA)-sensitive nuclear translocation after activation by xenobiotics such as phenobarbital (PB) and 1,4 bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP). We have now mimicked this TCPOBOP-dependent and OA-sensitive translocation of mouse CAR (mCAR) in HepG2 cells and have demonstrated that protein phosphatase 2A regulates this nuclear translocation. Site-directed mutagenesis analysis of various Ser and Thr residues delineated the translocation activity to Ser-202. Mutation of Ser-202 to Asp (S202D) prevented mCAR translocation into the nucleus of TCPOBOP-treated HepG2 cells. In addition, in the livers of Car-/- mice, the YFP-tagged S202D mutant did not translocate into the nucleus after PB treatment. To examine whether Ser-202 can be phosphorylated, flag-tagged wild-type mCAR or flag-tagged S202A mutant was expressed in HepG2 cells and subjected to Western blot analysis using an antibody specific to a peptide containing phospho-Ser-202. A high molecular weight phosphorylated form of CAR was detected only with the wild-type mCAR. These results are consistent with the conclusion that the dephosphorylation of Ser-202 is a required step that regulates the xenobiotic-dependent nuclear translocation of mCAR. PMID- 16377763 TI - Oral benzo[a]pyrene in Cyp1 knockout mouse lines: CYP1A1 important in detoxication, CYP1B1 metabolism required for immune damage independent of total body burden and clearance rate. AB - CYP1A1 and CYP1B1 metabolically activate many polycyclic aromatic hydrocarbons (PAHs), including benzo[a]pyrene, to reactive intermediates associated with toxicity, mutagenesis, and carcinogenesis. Paradoxically, however, Cyp1a1-/- knockout mice are more sensitive to oral benzo[a]pyrene exposure, compared with wild-type Cyp1a1+/+ mice (Mol Pharmacol 65:1225, 2004). To further investigate the mechanism for this enhanced sensitivity, Cyp1a1-/-, Cyp1a2-/-, and Cyp1b1-/- single-knockout, Cyp1a1/1b1-/- and Cyp1a2/1b1-/- double-knockout, and Cyp1+/+ wild-type mice were analyzed. After administration of oral benzo[a]pyrene (125 mg/kg/day) for 18 days, Cyp1a1-/- mice showed marked wasting, immunosuppression, and bone marrow hypocellularity, whereas the other five genotypes did not. After 5 days of feeding, steady-state blood levels of benzo[a]pyrene were approximately 25 and approximately 75 times higher in Cyp1a1-/- and Cyp1a1/1b1-/- mice, respectively, than in wild-type mice. Benzo[a]pyrene-DNA adduct levels were highest in liver, spleen, and marrow of Cyp1a1-/- and Cyp1a1/1b1-/- mice. Many lines of convergent data obtained with oral benzo[a]pyrene dosing suggest that: 1) inducible CYP1A1, probably in both intestine and liver, is most important in detoxication; 2) CYP1B1 in spleen and marrow is responsible for metabolic activation of benzo[a]pyrene, which results in immune damage in the absence of CYP1A1; 3) both thymus atrophy and hepatocyte hypertrophy are independent of CYP1B1 metabolism but rather may reflect long-term activation of the aryl hydrocarbon receptor; and 4) the magnitude of immune damage in Cyp1a1-/- and Cyp1a1/1b1-/- mice is independent of plasma benzo[a]pyrene and total-body burden and clearance. Thus, a balance between tissue-specific expression of the CYP1A1 and CYP1B1 enzymes governs sensitivity of benzo[a]pyrene toxicity and, possibly, carcinogenicity. PMID- 16377765 TI - Valproic acid increases conservative homologous recombination frequency and reactive oxygen species formation: a potential mechanism for valproic acid induced neural tube defects. AB - Valproic acid, a commonly used antiepileptic agent, is associated with a 1 to 2% incidence of neural tube defects when taken during pregnancy; however, the molecular mechanism by which this occurs has not been elucidated. Previous research suggests that valproic acid exposure leads to an increase in reactive oxygen species (ROS). DNA damage due to ROS can result in DNA double-strand breaks, which can be repaired through homologous recombination (HR), a process that is not error-free and can result in detrimental genetic changes. Because the developing embryo requires tight regulation of gene expression to develop properly, we propose that the loss or dysfunction of genes involved in embryonic development through aberrant HR may ultimately cause neural tube defects. To determine whether valproic acid induces HR, Chinese hamster ovary 3-6 cells, containing a neomycin direct repeat recombination substrate, were exposed to valproic acid for 4 or 24 h. A significant increase in HR after exposure to valproic acid (5 and 10 mM) for 24 h was observed, which seems to occur through a conservative HR mechanism. We also demonstrated that exposure to valproic acid (5 and 10 mM) significantly increased intracellular ROS levels, which were attenuated by preincubation with polyethylene glycol-conjugated (PEG)-catalase. A significant change in the ratio of 8-hydroxy-2'-deoxyguanosine/2'-de oxyguanosine, a measure of DNA oxidation, was not observed after valproic acid exposure; however, preincubation with PEG-catalase significantly blocked the increase in HR. These data demonstrate that valproic acid increases HR frequency and provides a possible mechanism for valproic acid-induced neural tube defects. PMID- 16377766 TI - Probing N-methyl-D-aspartate receptor desensitization with the substituted cysteine accessibility method. AB - Several forms of macroscopic N-methyl-D-aspartate (NMDA) receptor desensitization affect the amplitude and duration of postsynaptic responses. In addition to its functional significance, desensitization provides one means to examine the conformational coupling of ligand binding to channel gating. Segments flanking the ligand binding domain in the extracellular N terminus of the NMDA receptor NR2 subunit influence the glycine-independent form of desensitization. The NR2A pre-M1 region, the linker between the glutamate binding domain and the channel pore, plays a critical role in desensitization. Thus, we used the substituted cysteine accessibility method to scan the accessibility of residues in the pre-M1 region and the first transmembrane domain (M1) of NR2A. Cysteine mutants were expressed with NR1 in human embryonic kidney 293 cells and were assayed by whole cell recording. With activation of the receptor by glutamate and glycine, only a single mutant, V557C, which is located at the beginning of M1, led to irreversible inhibition by the methanethiosulfonate derivative methanethiosulfonate ethyltrimethylammonium (MTSET). The NR2 ligand glutamate was insufficient on its own to induce modification of V557C by MTSET, suggesting that the change in accessibility required channel gating. The rate of MTSET modification of the homologous residue on NR1 (NR1-1a(L562C)/NR2A) was much slower than V557C. We also substituted cysteine in the V557 site of mutant subunits that exhibit either enhanced or reduced desensitization. Modification by MTSET correlated with the degree of desensitization for these subunits, suggesting that V557C is a sensitive detector of desensitization gating. PMID- 16377767 TI - A systematic review of practice standards and research ethics in technology-based home health care intervention programs for older adults. AB - OBJECTIVE: The purpose of the review is to assess frequencies of reporting adherence to professional practice standards and research ethics in studies of technology-based home health care programs. METHODS: Key databases were searched to yield 2,866 abstracts that were independently rated by two reviewers using inclusion-exclusion criteria, resulting in 107 articles that were then reviewed for reports of practice standards and research ethics. RESULTS: Issues related to professional practice standards and research ethics were not well reported. When reported, adherence to practice standards included preintervention training, use of intervention protocols, supervision, and mechanisms for risk management. Research ethics most commonly reported were informed consent, REB/IRB approval, and protection of privacy. DISCUSSION: The results raise questions as to whether practice standards and research ethics are addressed sufficiently when health service delivery occurs in technology-based environments. Guidelines for professional accountability in e-health service delivery are needed. PMID- 16377768 TI - Community-based health coaching, exercise, and health service utilization. AB - OBJECTIVE: The outcomes of the California Public Employees Retirement System's Health Matters program, a replication of a health promotion and fitness program known as the Senior Wellness Program (SWP), are reported. METHOD: A randomized controlled design (n = 504) was used. Eligibility included one or more qualifying chronic health conditions, age 65 or older, member of a participating health plan, and owner of a CalPERS long-term care insurance policy. Disability risk factors were measured via questionnaires and included health status, pain, exercise, depression, and social activity. Activity was compiled in project records. RESULTS: At 12 months, Health Matters members were engaged in significantly more stretching and aerobic exercise than the controls. Depressive symptoms decreased among those with moderate or higher symptom scores. DISCUSSION: Health Matters extends the SWP model to younger, more active, and healthier individuals with improvements in duration of exercise and stretching; use of other health promotion activities was low. PMID- 16377769 TI - Older adults' and case managers' perceptions of smoking and smoking cessation. AB - OBJECTIVE: This study explores older smokers' and case managers' perceptions of smoking and smoking cessation. METHODS: Data were collected from 48 case managers via focus group sessions and questionnaires and from 20 smoking clients via structured interviews. RESULTS: Case managers identified barriers to addressing clients' smoking at the client, staff, and organizational levels. Clients and case managers lacked information on the benefits of smoking cessation in older adults. The majority of clients interviewed had no plans to quit smoking. DISCUSSION: Results suggest that smoking cessation programs for this population need to be geared to clients who are not ready to quit. Education about the benefits of quitting smoking in older adults is needed for both case managers and smokers. PMID- 16377770 TI - Are foot disorders associated with functional limitation and disability among community-dwelling older adults? AB - The objectives of this study are to examine whether specific foot disorders and ankle weakness and foot pain are related to functional limitations or disabilities in elders. Community-dwelling adults 65 and older were enrolled in a population-based, cross-sectional study of foot disorders and health outcomes. Demographics, health status, comorbidities, self-reported foot and knee pain, function and disability, and observed structural foot disorders, body mass index, and ankle muscle strength were assessed on 717 participants. The associations of foot disorders, foot pain, and ankle muscle weakness with function and disability were examined with regression analyses. Foot disorders were not associated with functional outcomes or disability. Ankle weakness was associated with performance based function (p = .005), self-report function (p lesser than .001), and disability (p = .009). Foot pain was associated with self-report function (p = .01) and disability (p = .007). Foot pain and ankle weakness seem to be related to important health outcomes among older adults. PMID- 16377771 TI - Intervening on exercise and nutrition in older adults: the Rhode Island SENIOR Project. AB - Presented are the basic design, methods, and baseline data analyses for the Study of Exercise and Nutrition in Older Rhode Islanders (SENIOR Project) an experimental study to investigate the relative effectiveness of a 12-month, stage of readiness to change-based multiple-behavior intervention (exercise and nutrition) compared to single-behavior interventions in a community-dwelling population of 1,277 older adults. Relationships between stage of readiness to change in the two target behaviors, as well as the relationship between stage of readiness and self-reported exercise levels and fruit and vegetable consumption, were examined using a combination of Pearson chi-squares, analyses of variance (ANOVA), and Spearman's rank order correlations. Stage of change (SOC) for fruit and vegetable consumption was significantly associated with the dietary measure, and SOC for exercise was associated with both the three physical activity measures and servings of fruits and vegetables per day. Overall, individual older adult's readiness to change seems largely to be behavior-specific. PMID- 16377772 TI - Gendered trajectories of work control and health outcomes in the middle years: a perspective from the rural Midwest. AB - OBJECTIVE: The objective was to investigate whether increasing health heterogeneity during the middle years is attributed, at least in part, to the influence of varying levels of, and changes in, work control of the current midlife cohort. METHOD: The study used four waves of data collected from 372 employed rural women and 320 employed men of the midlife cohort during a 10-year period. Variables included self-reported work control, stressful nonwork life events, and mental and physical health. The analyses used latent growth curve modeling. RESULTS: The results partially supported the hypothesized pathways. For middle-aged men, work control directly influences health outcomes, whereas for middle-aged women work control indirectly influences health outcomes through the occurrence of stressful life events. DISCUSSION: Through understanding these processes, work can be better designed to promote positive health outcomes, minimize negative health outcomes, and allow for better formulation and more effective implementation of health promotional programs. PMID- 16377773 TI - The social distribution of psychological distress and depression in older adults. AB - OBJECTIVE: This study examines the associations between social position and mental health and explores whether differences in distress and depression by social position can be accounted for by differences in the major components of the stress process model. We extend previous work by including an ethnocultural measure alongside more traditional measures of social position. METHOD: Secondary data analysis of the 1994 National Population Health Survey. RESULTS: Consistent with findings from studies of younger adults, mental health in later life is determined in part by age, gender, marital status, education, and ethnocultural factors. The data indicate that the life experiences connected to these social positions are largely responsible for these effects. DISCUSSION: Our findings suggest that key social factors are related to mental health in late life, because one's position in the social structure shapes the stressors they encounter and the resources they have at their disposal to cope with them. PMID- 16377774 TI - Predicting self-care behaviors among older adults coping with arthritis: a cross sectional and 1-year longitudinal comparative analysis. AB - OBJECTIVE: The purpose of this investigation is to examine correlates and predictors of self-care activities for persons diagnosed with arthritis both cross-sectionally and longitudinally. METHOD: Data from telephone surveys conducted with 313 older (M = 68.8, SD = 8.93) individuals, chosen from a larger sample, who reported professionally diagnosed arthritis, were used. RESULTS: A total of 10 of the 11 self-care activities changed significantly during the 1 year interval, with 9 showing increased participation. Results from hierarchical regressions showed that all three blocks of predictors explained significant portions of variance, with gender and perceived importance of general health significantly predicting self-care activities at Time 1, at Time 2, and longitudinally. DISCUSSION: These results highlight the influence of demographic, health status, and health belief variables on self-care both cross-sectionally and longitudinally. Future work should focus on the mediating effects of these and other variables to better understand the processes by which individuals engage in self-care behavior. PMID- 16377775 TI - Ineffectiveness of Lactobacillus johnsonii LA1 for prophylaxis of postoperative recurrence in Crohn's disease: a randomised, double blind, placebo controlled GETAID trial. AB - BACKGROUND AND AIMS: Early endoscopic recurrence is frequent after intestinal resection for Crohn's disease. Bacteria are involved, and probiotics may modulate immune responses to the intestinal flora. Here we tested the probiotic strain Lactobacillus johnsonii LA1 in this setting. PATIENTS AND METHODS: This was a randomised, double blind, placebo controlled study. Patients were eligible if they had undergone surgical resection of <1 m, removing all macroscopic lesions within the past 21 days. Patients were randomised to receive two packets per day of lyophilised LA1 (2 x 10(9) cfu) or placebo for six months; no other treatment was allowed. The primary endpoint was endoscopic recurrence at six months, with grade >1 in Rutgeerts' classification or an adapted classification for colonic lesions. Endoscopic score was the maximal grade of ileal and colonic lesions. Analyses were performed primarily on an intent to treat basis. RESULTS: Ninety eight patients were enrolled (48 in the LA1 group). At six months, endoscopic recurrence was observed in 30/47 patients (64%) in the placebo group and in 21/43 (49%) in the LA1 group (p = 0.15). Per protocol analysis confirmed this result. Endoscopic score distribution did not differ significantly between the LA1 and placebo groups. There were four clinical recurrences in the LA1 group and three in the placebo group. CONCLUSION: L johnsonii LA1 (4 x 10(9) cfu/day) did not have a sufficient effect, if any, to prevent endoscopic recurrence of Crohn's disease. PMID- 16377776 TI - Use of a mixed tissue RNA design for performance assessments on multiple microarray formats. AB - The comparability and reliability of data generated using microarray technology would be enhanced by use of a common set of standards that allow accuracy, reproducibility and dynamic range assessments on multiple formats. We designed and tested a complex biological reagent for performance measurements on three commercial oligonucleotide array formats that differ in probe design and signal measurement methodology. The reagent is a set of two mixtures with different proportions of RNA for each of four rat tissues (brain, liver, kidney and testes). The design provides four known ratio measurements of >200 reference probes, which were chosen for their tissue-selectivity, dynamic range coverage and alignment to the same exemplar transcript sequence across all three platforms. The data generated from testing three biological replicates of the reagent at eight laboratories on three array formats provides a benchmark set for both laboratory and data processing performance assessments. Close agreement with target ratios adjusted for sample complexity was achieved on all platforms and low variance was observed among platforms, replicates and sites. The mixed tissue design produces a reagent with known gene expression changes within a complex sample and can serve as a paradigm for performance standards for microarrays that target other species. PMID- 16377777 TI - Solution structure of psi32-modified anticodon stem-loop of Escherichia coli tRNAPhe. AB - Nucleoside base modifications can alter the structures and dynamics of RNA molecules and are important in tRNAs for maintaining translational fidelity and efficiency. The unmodified anticodon stem-loop from Escherichia coli tRNA(Phe) forms a trinucleotide loop in solution, but Mg2+ and dimethylallyl modification of A37 N6 destabilize the loop-proximal base pairs and increase the mobility of the loop nucleotides. The anticodon arm has three additional modifications, psi32, psi39, and A37 C2-thiomethyl. We have used NMR spectroscopy to investigate the structural and dynamical effects of psi32 on the anticodon stem-loop from E.coli tRNA(Phe). The psi32 modification does not significantly alter the structure of the anticodon stem-loop relative to the unmodified parent molecule. The stem of the RNA molecule includes base pairs psi32-A38 and U33-A37 and the base of psi32 stacks between U33 and A31. The glycosidic bond of psi32 is in the anti configuration and is paired with A38 in a Watson-Crick geometry, unlike residue 32 in most crystal structures of tRNA. The psi32 modification increases the melting temperature of the stem by approximately 3.5 degrees C, although the psi32 and U33 imino resonances are exchange broadened. The results suggest that psi32 functions to preserve the stem integrity in the presence of additional loop modifications or after reorganization of the loop into a translationally functional conformation. PMID- 16377778 TI - Identification of a 14mer RNA that recognizes and binds flavin mononucleotide with high affinity. AB - Aptamers are nucleic acids developed by in vitro evolution techniques that bind to specific ligands with high affinity and selectivity. Despite such high affinity and selectivity, however, in vitro evolution does not necessarily reveal the minimum structure of the nucleic acid required for selective ligand binding. Here, we show that a 35mer RNA aptamer for the cofactor flavin mononucleotide (FMN) identified by in vitro evolution can be computationally evolved to a mere 14mer structure containing the original binding pocket and eight scaffolding nucleotides while maintaining its ability to bind in vitro selectively to FMN. Using experimental and computational methodologies, we found that the 14mer binds with higher affinity to FMN (K(D) approximately 4 microM) than to flavin adenine dinucleotide (K(D) approximately 12 microM) or to riboflavin (K(D) approximately 13 microM),despite the negative charge of FMN. Different hydrogen-bond strengths resulting from differing ring-system electron densities associated with the aliphatic-chain charges appear to contribute to the selectivity observed for the binding of the 14mer to FMN and riboflavin. Our results suggest that high affinity and selectivity in ligand binding is not restricted to large RNAs, but can also be a property of extraordinarily short RNAs. PMID- 16377779 TI - Oligomerization of HEXIM1 via 7SK snRNA and coiled-coil region directs the inhibition of P-TEFb. AB - Transcriptional elongation of most eukaryotic genes by RNA polymerase II requires the kinase activity of the positive transcription elongation factor b (P-TEFb). The catalytically active P-TEFb complex becomes inactive when sequestered into the large complex by the cooperative actions of 7SK snRNA and HEXIM1. In this study, we report that HEXIM1 forms oligomers in cells. This oligomerization is mediated by its predicted coiled-coil region in the C-terminal domain and 7SK snRNA that binds a basic region within the central part of HEXIM1. Alanine mutagenesis of evolutionary conserved leucines in the coiled-coil region and the digestion of 7SK snRNA by RNase A treatment prevent this oligomerization. Importantly, mutations of the N-terminal part of the coiled-coil region abrogate the ability of HEXIM1 to bind and inhibit P-TEFb. Finally, the formation of HEXIM1 oligomers via the C-terminal part of the coiled-coil or basic regions is critical for the inhibition of transcription. Our results suggest that two independent regions in HEXIM1 form oligomers to incorporate P-TEFb into the large complex and determine the inhibition of transcriptional elongation. PMID- 16377780 TI - Synthesis and hybridization studies of oligonucleotides containing 1-(2-deoxy-2 alpha-C-hydroxymethyl-beta-D-ribofuranosyl)thymine (2'-alpha-hm-dT). AB - We report the first investigation of oligoribonucleotides containing a few 1-(2 deoxy-2-alpha-C-hydroxymethyl-beta-D-ribofuranosyl)thymine units (or 2'-hm-dT, abbreviated in this work as 'H'). Both the 2'-CH2O-phosphoramidite and 3'-O phosphoramidite derivatives of H were synthesized and incorporated into both 2',5'-RNA and RNA chains. The hybridization properties of the modified oligonucleotides have been studied via thermal denaturation and circular dichroism studies. While 3',5'-linked H was shown previously to significantly destabilize DNA:RNA hybrids and DNA:DNA duplexes (modification in the DNA strand; DeltaT(m) approximately -3 degrees C/insert), we find that 2',5'-linked H have a smaller effect on 2',5'-RNA:RNA and RNA:RNA duplexes (DeltaT(m) = -0.3 degrees C and -1.2 degrees C, respectively). The incorporation of 3',5'-linked H into 2',5' RNA:RNA and RNA:RNA duplexes was found to be more destabilizing (-0.7 degrees C and -3.6 degrees C, respectively). Significantly, however, the 2',5'-linked H units confer marked stability to RNA hairpins when they are incorporated into a 2',5'-linked tetraloop structure (DeltaT(m) = +1.5 degrees C/insert). These results are rationalized in terms of the compact and extended conformations of nucleotides. PMID- 16377782 TI - Bovine tuberculosis: DEFRA consults on badger culling. PMID- 16377781 TI - Easily denaturing nucleic acids derived from intercalating nucleic acids: thermal stability studies, dual duplex invasion and inhibition of transcription start. AB - The bulged insertions of (R)-1-O-(pyren-1-ylmethyl)glycerol (monomer P) in two complementary 8mer DNA strands (intercalating nucleic acids) opposite to each other resulted in the formation of an easily denaturing duplex, which had lower thermal stability (21.0 degrees C) than the wild-type double-stranded DNA (dsDNA, 26.0 degrees C), but both modified oligodeoxynucleotides had increased binding affinity toward complementary single-stranded DNA (ssDNA) (41.5 and 39.0 degrees C). Zipping of pyrene moieties in an easily denaturing duplex gave formation of a strong excimer band at 480 nm upon excitation at 343 nm in the steady-state fluorescence spectra. The excimer band disappeared upon addition of a similar short dsDNA, but remained when adding a 128mer dsDNA containing the same sequence. When P was inserted into 2'-OMe-RNA strands, the duplex with zipping P was found to be more stable (42.0 degrees C) than duplexes with the complementary ssDNAs (31.5 and 19.5 degrees C). The excimer band observed in the ds2'-OMe-RNA with zipping P had marginal changes upon addition of both 8 and 128mer dsDNA. Synthesized oligonucleotides were tested in a transcriptional inhibition assay for targeting of the open complex formed by Escherichia coli RNA polymerase with the lac UV-5 promoter using the above mentioned 128mer dsDNA. Inhibition of transcription was observed for 8mer DNAs possessing pyrene intercalators and designed to target both template and non-template DNA strands within the open complex. The observed inhibition was partly a result of unspecific binding of the modified DNAs to the RNA polymerase. Furthermore, the addition of 8mer DNA with three bulged insertions of P designed to be complementary to the template strand at the +36 to +43 position downstream of the transcription start resulted in a specific halt of transcription producing a truncated RNA transcript. This is to our knowledge the first report of an RNA elongation stop mediated by a small DNA sequence possessing intercalators. The insertions of P opposite to each other in ds2'-OMe-RNA showed inhibition efficiency of 96% compared with 25% for unmodified ds2'-OMe-RNA. PMID- 16377785 TI - Veterinary strategy for the new farm health planning initiative. PMID- 16377787 TI - Complications observed during and after ovariohysterectomy of 142 bitches at a veterinary teaching hospital. AB - The clinical, anaesthetic and surgical records of 142 bitches admitted into the primary care clinic at the Small Animal Hospital, University of Liverpool, between January 2002 and August 2004 for routine ovariohysterectomy by final-year students were analysed for complications during and after the surgery. The time taken to perform the surgical procedure was recorded, to allow assessment of the time involved in one-to-one surgical teaching during live-recovery surgeries of client-owned pets. The types of complications were similar to those reported in earlier studies: nine bitches suffered haemorrhage of the ovarian arteries during surgery, and there were four cases of haemorrhage after surgery, 12 cases of wound inflammation and four other types of complication. The rates of intraoperative, postoperative and total complications were 6.3, 14.1 and 20.6 per cent, respectively. PMID- 16377788 TI - Immunopathological investigations on bovine digital epidermitis. AB - Paraffin-embedded fragments of bovine digital skin lesions were sectioned and stained with Warthin-Starry, haematoxylin and eosin, Grocott's methenamine silver and immunohistochemical techniques. Microorganisms observed in the silver-stained sections were classified into four major morphological groups. Spirochaetes were the most prevalent organisms, but bacillary and coccoid elements were also present in most sections. Immunohistochemical probing demonstrated that approximately 80 per cent, 46 per cent and 41 per cent of the digital and interdigital dermatitis sections stained positively with polyclonal antisera to Treponema pallidum, Campylobacter jejuni and Fusobacterium necrophorum, respectively. An unidentified branching filamentous organism (presumed to be an actinomycete) was consistently present in the sections of samples from mild interdigital lesions. PMID- 16377789 TI - Impact of the duration of control of cattle lice with eprinomectin on leather quality. AB - The relationship between lice infestation in calves during their first winter and damage to the leather produced was investigated in a trial involving 500 calves, 100 of which were treated with a pour-on endectocide during the first winter. All the calves received routine lice treatment in the second winter and were reared to slaughter weight. The hides were removed at the abattoir, tanned, inspected for lice-related damage, and graded according to their suitability for the production of high quality leather. The untreated group developed natural infestations of the chewing louse (Bovicola bovis) during the first winter but none was observed in the second winter. Hides from cattle infested with lice in their first winter had higher levels of lice damage than hides from those treated with eprinomectin, at both the chrome-tanned and dried dyed-crust stages of leather production. PMID- 16377790 TI - Normal haematological values in gyr falcons (Falco rusticolus). PMID- 16377791 TI - Malignant peripheral nerve sheath tumour of the heart in a horse. PMID- 16377792 TI - Unusual encephalopathy in weaned lambs. PMID- 16377793 TI - BVA/KC hip and elbow dysplasia schemes. PMID- 16377794 TI - Strengthening the requirement for CPD. PMID- 16377796 TI - Assessing the threat of avian influenza. PMID- 16377798 TI - Pulmonary artery haemorrhages in sheep with septicaemic pasteurellosis. PMID- 16377799 TI - PTPN11 (Shp2) mutations in LEOPARD syndrome have dominant negative, not activating, effects. AB - Multiple lentigines/LEOPARD syndrome (LS) is a rare, autosomal dominant disorder characterized by Lentigines, Electrocardiogram abnormalities, Ocular hypertelorism, Pulmonic valvular stenosis, Abnormalities of genitalia, Retardation of growth, and Deafness. Like the more common Noonan syndrome (NS), LS is caused by germ line missense mutations in PTPN11, encoding the protein tyrosine phosphatase Shp2. Enzymologic, structural, cell biological, and mouse genetic studies indicate that NS is caused by gain-of-function PTPN11 mutations. Because NS and LS share several features, LS has been viewed as an NS variant. We examined a panel of LS mutants, including the two most common alleles. Surprisingly, we found that in marked contrast to NS, LS mutants are catalytically defective and act as dominant negative mutations that interfere with growth factor/Erk-mitogen-activated protein kinase-mediated signaling. Molecular modeling and biochemical studies suggest that LS mutations contort the Shp2 catalytic domain and result in open, inactive forms of Shp2. Our results establish that the pathogenesis of LS and NS is distinct and suggest that these disorders should be distinguished by mutational analysis rather than clinical presentation. PMID- 16377800 TI - Hepatocyte nuclear factor-4alpha is essential for glucose-stimulated insulin secretion by pancreatic beta-cells. AB - Mutations in the hepatocyte nuclear factor (HNF)-4alpha gene cause a form of maturity-onset diabetes of the young (MODY1) that is characterized by impairment of glucose-stimulated insulin secretion by pancreatic beta-cells. HNF-4alpha, a transcription factor belonging to the nuclear receptor superfamily, is expressed in pancreatic islets as well as in the liver, kidney, and intestine. However, the role of HNF-4alpha in pancreatic beta-cell is unclear. To clarify the role of HNF 4alpha in beta-cells, we generated beta-cell-specific HNF-4alpha knock-out (betaHNF-4alphaKO) mice using the Cre-LoxP system. The betaHNF-4alphaKO mice exhibited impairment of glucose-stimulated insulin secretion, which is a characteristic of MODY1. Pancreatic islet morphology, beta-cell mass, and insulin content were normal in the HNF-4alpha mutant mice. Insulin secretion by betaHNF 4alphaKO islets and the intracellular calcium response were impaired after stimulation by glucose or sulfonylurea but were normal after stimulation with KCl or arginine. Both NAD(P)H generation and ATP content at high glucose concentrations were normal in the betaHNF-4alphaKO mice. Expression levels of Kir6.2 and SUR1 proteins in the betaHNF-4alphaKO mice were unchanged as compared with control mice. Patch clamp experiments revealed that the current density was significantly increased in betaHNF-4alphaKO mice compared with control mice. These results are suggestive of the dysfunction of K(ATP) channel activity in the pancreatic beta-cells of HNF-4alpha-deficient mice. Because the K(ATP) channel is important for proper insulin secretion in beta-cells, altered K(ATP) channel activity could be related to the impaired insulin secretion in the betaHNF 4alphaKO mice. PMID- 16377801 TI - Silent or non-clinical infarct-like lesions in the posterior circulation territory in migraine: brain hypoperfusion or hyperperfusion? PMID- 16377804 TI - New advances in the pathophysiology of focal dystonias. PMID- 16377805 TI - Epigenetic silencing of E- and N-cadherins in the stroma of mouse thymic lymphomas. AB - Aberrant expression of some tumour suppressor genes and oncogenes by thymocytes had been involved in the development of primary thymic lymphomas induced by gamma irradiation, but genetic alterations affecting critical genes expressed by stromal cells have not been yet explored. This paper analyzes a series of such tumours induced in C57BL/6J and in F1 hybrids of BALB/c and C57BL/6J mouse strains. As expected, hystopathological analyses revealed profound disorganizations within the thymus with a poor demarcation of the cortical and medullar areas. Immunological and quantitative on-line RT-PCR analyses confirm that E-cadherin (Cdh1) is essentially expressed by stromal cells of the thymus, while evidencing that the expression of this gene is significantly reduced in all tumours. In addition, and contrary to what one would expect, N-cadherin (Cdh2) that is exclusively expressed by stromal cells is likewise down-regulated in most of the thymic lymphomas. Although hypermethylation of the promoter region appears to be involved in the inactivation of Cdh2 in all tumours, additional epigenetic mechanisms mediated by repressors such as Snai1 may also play a role in Cdh1 silencing. These results represent the first reported case for tumour-associated gene alterations occurring not in the tumour cells per se, but in the stromal cells of primary thymic lymphomas. Additionally, since the expression of both genes is significantly up-regulated after a single high dose of gamma-radiation, but remained unchanged in treated thymic-lymphoma-free-mice, epigenetic down regulation of E- and N-cadherin appears to occur concomitantly with the progression towards the most advanced stages of gamma-radiation-induced thymic lymphomas. PMID- 16377806 TI - Differential susceptibility of mice humanized for peroxisome proliferator activated receptor alpha to Wy-14,643-induced liver tumorigenesis. AB - Peroxisome proliferators, such as lipid-lowering fibrate drugs, are agonists for the peroxisome proliferator-activated receptor alpha (PPARalpha). Sustained activation of PPARalpha leads to the development of liver tumors in rodents. Paradoxically, humans appear to be resistant to the induction of peroxisome proliferation and development of liver tumors by peroxisome proliferators. To examine the species differences in response to peroxisome proliferators, a PPARalpha humanized mouse (hPPARalpha) was generated, in which the human PPARalpha was expressed in liver under control of the Tet-OFF system. To evaluate the susceptibility of hPPARalpha mice to peroxisome proliferator-induced hepatocarcinogenesis, a long-term feeding study of Wy-14,643 was carried out. hPPARalpha and wild-type (mPPARalpha) mice were fed either a control diet or one containing 0.1% Wy-14,643 for 44 and 38 weeks, respectively. Gene expression analysis for peroxisomal and mitochondrial fatty acid metabolizing enzymes revealed that both hPPARalpha and mPPARalpha were functional. However, the incidence of liver tumors including hepatocellular carcinoma was 71% in Wy-14,643 treated mPPARalpha mice, and 5% in Wy-14,643-treated hPPARalpha mice. Upregulation of cell cycle regulated genes such as cd1 and Cdks were observed in non-tumorous liver tissue of Wy-14,643-treated mPPARalpha mice, whereas p53 gene expression was increased only in the livers of Wy-14,643-treated hPPARalpha mice. These findings suggest that structural differences between human and mouse PPARalpha are responsible for the differential susceptibility to the peroxisome proliferator-induced hepatocarcinogenesis. This mouse model will be useful for human cancer risk assessment of PPARalpha ligands. PMID- 16377807 TI - MLL rearrangements are induced by low doses of etoposide in human fetal hematopoietic stem cells. AB - During fetal development, the liver serves as the primary hematopoietic organ in which hematopoietic stem cells (HSC) comprise a large proportion of hepatic cell populations. Because HSC are capable of initiating long-term hematopoiesis, injury to these cells during pregnancy may play a role in the development of hematopoietic disorders manifested after birth. Of interest is the role of genetic injury to fetal HSC in the etiology of the infant acute leukemias, which are characterized by chromosomal rearrangements in the 11q23 region involving the mixed lineage leukemia (MLL) gene. These gene fusions also occur in leukemias in adults following chemotherapy with etoposide and other inhibitors of DNA topoisomerase II. We used etoposide as a model compound to determine the sensitivity of human fetal HSC to DNA damage and to determine whether we could induce MLL rearrangements in cultured human fetal HSC. Exposure of HSC to etoposide resulted in a dose-dependent loss of viability, with effects observed at low nanomolar concentrations. DNA strand breaks were observed on exposure to 140 nM etoposide, and higher etoposide concentrations stimulated an increase in early lymphoid populations and elicited G2/M cell cycle arrest. Immunophenotyping of MLL translocations revealed a significant increase in positive flow cytometry events at low etoposide concentrations and were consistent with MLL recombination. MLL translocations were confirmed using fluorescent in situ hybridization. In vitro inhibition of DNA topoisomerase II was observed at >or=25 microM etoposide, but was not evident at lower etoposide concentrations associated with DNA damage. Our data indicate that low acute doses of etoposide can cause DNA strand breaks and chromosomal rearrangements involving MLL in human fetal HSC. Ultimately, such injury may have ramifications with regards to transplacental exposures to environmental chemicals linked to the etiology of infant acute leukemias. PMID- 16377808 TI - To better serve the interests of patients. PMID- 16377812 TI - Characteristics, roles, and responsibilities of the Designated Institutional Official (DIO) position in graduate medical education. AB - PURPOSE: In 1998, the Accreditation Council for Graduate Medical Education (ACGME) added the stipulation that each institution providing graduate medical education (GME) have a Designated Institutional Official (DIO). Little is known about the effect of new accreditation requirements on GME practice and outcomes. The authors conducted a cross-sectional survey designed to provide descriptive data about DIOs and to validate a DIO Responsibility Scale (DIORS). METHOD: DIOs were identified by the ACGME. The following delivery strategy was used to administer the survey from January 2004 to May 2004: prenotice letter; survey with self-addressed, stamped return envelope and cover letter; thank-you/reminder postcard; and replacement survey with new cover letter and self-addressed, stamped return envelope. RESULTS: Completed surveys were received from 243 of 363 DIOs (66.9%). Responses indicated wide ranges in DIO titles, report titles, time spent accomplishing responsibilities, DIO-specific salaries, credentials, and Graduate Medical Education Committee reporting arrangements. DIOs reported confusion or overlap between DIO and program director roles (72.0%), decreasing funding for GME (50.6%), and inadequate support staff (25.1%). The 11-item DIO Responsibility Scale demonstrated a Cronbach alpha of .86 and a statistically significant relationship to five variables selected to establish construct validity. CONCLUSIONS: The wide variability in DIO characteristics found in this study supports the premise that the DIO role is underdeveloped. The authors established the reliability and validity of the DIORS, which now may be used in future research of the DIO role. The data from this study may be used by DIOs and institutions to develop the role further, improve DIO performance, and create more useful job descriptions. PMID- 16377814 TI - Leadership values in academic medicine. AB - PURPOSE: To gain a deeper understanding of the guiding core values that deans of academic medical centers (AMCs) considered most essential for their leadership and the major leadership challenges that confront them. METHOD: In 2003-04, semistructured interviews of 18 deans at U.S. colleges of medicine or AMCs were organized around four dimensions: background, leadership challenges, organizational effectiveness, and systems enablers/restrainers for leadership. A values Q-sort was used to determine how widely core values were shared among deans and how the complex challenges they faced did or did not align with these values. RESULTS: Fourteen of the 18 (78%) deans identified financial difficulties as their most pressing leadership challenge, followed by weak institutional alignment (61%), staffing problems (33%), and poor morale (28%). Open, candid communication was reported as the most effective means of addressing these complex problems. Enacting espoused shared values and having a positive attitude were identified as the most important enablers of systemic leadership, whereas micromanagement and difficult people were the major restraints. Q-sort results on 38 positive leadership values indicated that participants considered integrity most essential. Integrity was positively correlated with humanistic values and negatively correlated with results. Vision, another highly espoused value, correlated strongly with performance-oriented values but correlated negatively with humanistic values. CONCLUSIONS: A dynamic tension exists in AMCs between humanistic values and performance-based core values. The ability to manage that tension (i.e., when to prioritize one set of values over the other) is inherent in a dean's work. PMID- 16377815 TI - The impact of the changing health care environment on the health and well-being of faculty at four medical schools. AB - PURPOSE: Increased pressure for clinical and research productivity and decreased control over the work environment have been reported to have adverse impacts on academic faculty in limited studies. The authors examined whether work-related stressors in academic medicine negatively affected the physical and mental health, as well as life and job satisfaction, of academic medical school faculty. METHOD: A 136-item self-administered anonymous questionnaire modified from a small 1984 study was distributed to 3,519 academic faculty at four U.S. medical schools following institutional review board approval at each school. Validated scales measuring depression, anxiety, work strain, and job and life satisfaction; a checklist of common physical and mental health symptoms; and questions about the impact of institutional financial stability, colleague attrition, and other work-related perceptions were used. Responses were analyzed by sex, academic rank, age, marital status, faculty discipline, and medical school. RESULTS: Responses were received from 1,951 full-time academic physicians and basic science faculty, a 54.3% response rate. Twenty percent of faculty, almost equal by sex, had significant levels of depressive symptoms, with higher levels in younger faculty. Perception of financial instability was associated with greater levels of work strain, depression, and anxiety. Significant numbers of faculty acknowledged that work-related strain negatively affected their mental health and job satisfaction, but not life satisfaction or physical health. Specialties were differentially affected. CONCLUSIONS: High levels of depression, anxiety, and job dissatisfaction-especially in younger faculty-raise concerns about the well-being of academic faculty and its impact on trainees and patient care. Increased awareness of these stressors should guide faculty support and development programs to ensure productive, stable faculty. PMID- 16377816 TI - The physician-hospital team: a successful approach to improving care in a large academic medical center. AB - Initiatives to improve the quality and efficiency of care in academic medical centers (AMCs, teaching hospitals) can benefit the performance of academic departments as well as the hospital. However, the value of performance improvement programs in an AMC is often challenging. At Jefferson Medical College, clinical efficiency and bed availability are important priorities to the Department of Medicine. To this end, a multidisciplinary program was designed to (1) improve the quality and consistency of care by adapting and adopting national guidelines for patients with heart failure and acute coronary syndrome; (2) identify and improve hospital operational supports and maximize resource utilization; (3) increase hospital functional capacity to make way for increased volume; and (4) improve housestaff education and practice by using evidence-based approaches and by optimizing teaching relationships between housestaff and attending faculty. The eight-month project (November 2002 to July 2003) resulted in improvement in several quality measures including increased use of beta blockers and angiotensin converting enzyme inhibitors for heart failure patients, reduced length of stay for heart failure and acute coronary syndrome patients, and increased satisfaction of the clinicians involved in caring for these patients. However, the project was not without barriers including individual physician's unwillingness to embrace change and an inability to incentivize change. Development of faculty leadership skills and enhanced physician accountability helped in overcoming the challenges of change. PMID- 16377818 TI - Medicine and the arts. My Machine by Jack Coulehan, MD. Commentary. PMID- 16377817 TI - Academic research record-keeping: best practices for individuals, group leaders, and institutions. AB - During the last half of the 20th century, social and technological changes in academic research groups have challenged traditional research record-keeping practices, making them either insufficient or obsolete. New practices have developed but standards (best practices) are still evolving. Based on the authors' review and analysis of a number of sources, they present a set of systematically compiled best practices for research record-keeping for academic research groups. These best practices were developed as an adjunct to a research project on research ethics aimed at examining the actual research record-keeping practices of active academic scientists and their impact on research misconduct inquiries. The best practices differentiate and provide separate standards for three different levels within the university: the individual researcher, the research group leader, and the department/institution. They were developed using a combination of literature reviews, surveys of university integrity officials, focus groups of active researchers, and inspection of university policies on research record-keeping. The authors believe these best practices constitute a ''snapshot'' of the current normative standards for research records within the academic research community. They are offered as ethical and practical guidelines subject to continuing evolution and not as absolute rules. They may be especially useful in training the next generation of researchers. PMID- 16377820 TI - Duty-hour restrictions and the work of surgical faculty: results of a multi institutional study. AB - PURPOSE: To examine whether duty-hour restrictions have been consequential for various aspects of the work of surgical faculty and if those consequences differ for faculty in academic and nonacademic general surgery residency programs. METHOD: Questionnaires were distributed in 2004 to 233 faculty members in five academic and four nonacademic U.S. residency programs in general surgery. Participation was restricted to those who had been faculty for at least one year. Ten items on the questionnaire probed faculty work experiences. Results include means, percentages, and t-tests on mean differences. Of the 146 faculty members (63%) who completed the questionnaire, 101 volunteered to be interviewed. Of these, 28 were randomly chosen for follow-up interviews that probed experiences and rationales underlying items on the questionnaire. Interview transcripts (187 single-spaced pages) were analyzed for main themes. RESULTS: Questionnaire respondents and interviewees associated duty-hour restrictions with lowered faculty expectations and standards for residents, little change in the supervision of residents, a loss of time for teaching, increased work and stress, and less satisfaction. No significant differences in these perceptions (p < or = .05) were found for faculty in academic and nonacademic programs. Main themes from the interviews included a shift of routine work from residents to faculty, a transfer of responsibility to faculty, more frequent skill gaps at night, a loss of time for research, and the challenges of controlling residents' hours. CONCLUSIONS: Duty-hour restrictions have been consequential for the work of surgical faculty. Faculty should not be overlooked in future studies of duty-hour restrictions. PMID- 16377821 TI - Hand in hand. PMID- 16377822 TI - Multidimensional effects of the 80-hour work week at the University of Michigan Medical School. AB - PURPOSE: To examine the effects of the new resident work-hour restrictions on medical students, as measured by their perceptions of the quality of their experiences during the required clerkships. METHOD: Evaluations of four clerkships were compared for two student cohorts at the University of Michigan Medical School. The first cohort, from the class of 2002-03, completed their clinical clerkships the year before the work-hour restrictions were implemented, and the second cohort, from the class of 2003-04, completed their clerkships the same year the restrictions were implemented. RESULTS: There were significant and notable differences in the experiences of the two cohorts. Students' perceptions of the quality of their experiences in the surgery-oriented clerkships (obstetrics-gynecology and surgery) in particular were significantly lower (i.e., more negative) in the 2003-04 cohort than in the previous cohort for the same clerkships. The nonsurgery-oriented clerkships (internal medicine and pediatrics) hired hospitalists, who offset the residents' workload (internal medicine) and assumed teaching responsibilities (pediatrics). Between 2002-03 and 2003-04, students' perceptions of the quality of their experience in the internal medicine clerkship remained mostly stable, and increased in several areas for the students in the pediatrics clerkship. CONCLUSIONS: Implementation of resident work-hour restrictions had significant effects on the education of the medical students studied. These effects need to be carefully analyzed and considered to ensure quality education for medical students. The findings also highlight that the nature of students' perceptions was related to preparations made (or not) by specific clerkships as restricted work-hour regulations were adopted. PMID- 16377823 TI - Residents' perceptions of the effects of work hour limitations at a large teaching hospital. AB - PURPOSE: To assess residents' perceptions of the impact of resident work hour restrictions on patient care, education, and job satisfaction. METHOD: Four focus groups of internal medicine residents at Barnes-Jewish Hospital at the Washington University School of Medicine were conducted during February and March 2004. Twenty-six housestaff from the first three years of residency participated; all were volunteers. Transcripts were analyzed for major themes. RESULTS: Both residents and interns supported work hour limitations and enjoyed the benefits of working fewer hours. However, they had difficulty complying with the restrictions, particularly if they felt patient care, teaching, or their own education would be compromised. Participants perceived that restricted work hours diminished the continuity of patient care and increased the likelihood of medical errors such as those resulting from delayed follow-up of diagnostic tests. Both interns and residents found it difficult to attend conferences, and residents found fewer opportunities to teach. Effects on job satisfaction were mixed as a result of work hour restrictions. CONCLUSIONS: Residents in the sample favored work hour restrictions but had serious concerns about the effects of the restrictions on patient care and medical education. The findings suggest that imposing rigid work hour restrictions has significant consequences for patient care and medical education and that the most effective ways to balance work hour limitations with the demands of patient care and necessary educational components to train competent physicians have yet to be identified. PMID- 16377824 TI - Implementing duty-hour restrictions without diminishing patient care or education: can it be done? AB - PURPOSE: To implement and evaluate a new ward team call system that would meet the Accreditation Council on Graduate Medical Education's (ACGME) duty-hour requirements without compromising patient care or detracting from resident education. METHOD: The new system was implemented in the internal medicine residency program at the University Hospital at the University of Cincinnati Medical Center. In 2003-04, residents and faculty were surveyed about their experiences with the new and old ward systems relative to duty-hour requirements, patient care, and resident education. Responses were given on a five-point scale (5 = strongly agree, 1 = strongly disagree). Data were compiled and compared using a two-sample t-test. RESULTS: Faculty believed the new system improved compliance with the duty-hour requirements (mean = 4.3, 95% confidence interval [CI]: 4.1-4.6), although were neutral regarding patient care (mean = 3.5, 95% CI: 3.2-3.8) and education (mean = 3.3, 95% CI: 2.9-3.6). Residents were more neutral regarding ACGME requirements (mean = 3.5, 95% CI: 3.3-3.7) and patient care (mean = 3.2, 95% CI 3.0-3.3). Residents reported a slightly negative impact on education (mean = 2.8, 95% CI: 2.5-3.0). In response to an exclusive question, residents reported that the new system did not reduce fatigue (mean = 2.7, 95% CI: 2.6-3.0). CONCLUSIONS: Respondents perceived that this ward call system met ACGME requirements and maintained quality patient care but may have sacrificed some traditional resident education tenets. PMID- 16377825 TI - Impact of reduced duty hours on residents' educational satisfaction at the University of California, San Francisco. AB - PURPOSE: To assess the impact of the Accreditation Council for Graduate Medical Education duty-hour limitations on residents' educational satisfaction. METHOD: In 2003, the authors surveyed 164 internal medicine residents at three clinical training sites affiliated with the University of California, San Francisco, after system changes were introduced to reduce duty hours. On a questionnaire that used various rating scales, residents reported the value of educational activities, frequency of administrative tasks interfering with education, and educational satisfaction after duty hours were reduced. The authors compared univariate statistics and developed multivariable models to discern the relationship between hours worked and educational outcomes. RESULTS: In all, 125 residents (76%) responded. Residents rated the educational activities, morning report, and teaching others most highly. Answering pages and tasks related to scheduling were the most frequent barriers to educational activities. Residents reported that time spent in administrative activities did not change after duty-hour restrictions, and 68% said that decreased duty hours had no impact or a negative impact on education. In multivariable models, postgraduate year (PGY)-1 residents (p = .004), residents who reported feeling overwhelmed at work (p < .0001), and residents who reported working more than 80 hours per week (p < .05) had lower work satisfaction. However, only PGY-1 residents (p < .05) and those who felt overwhelmed with work (p = .01) were less satisfied with their education. CONCLUSIONS: In this residency program, duty-hour reduction did not improve educational satisfaction. Educational satisfaction may be more a function of workload than hours worked; therefore, systematic changes to residents' work-life may be necessary to improve educational satisfaction. PMID- 16377826 TI - Evolution of sleep quantity, sleep deprivation, mood disturbances, empathy, and burnout among interns. AB - PURPOSE: To explore the relationships between sleep deprivation and the evolution of mood disturbances, empathy, and burnout among a cohort of interns. METHOD: In 2002-03, 47 interns in the internal medicine resident program at the University of Pennsylvania School of Medicine completed the following instruments at baseline and at year end: sleep quantities, Epworth Sleepiness Scale, the Beck Depression Inventory-Short Form, the Interpersonal Reactivity Index, and the Maslach Burnout Inventory-Human Services Survey. The prevalences of acute and chronic sleep deprivation, subjective sleepiness, burnout, empathy, and depression at the beginning of the year were compared to prevalences at the end of internship. Associations between sleep deprivation and mood, empathy, or burnout were explored. RESULTS: The prevalence of chronic sleep deprivation, depression, burnout, and empathy increased from baseline to year end. Specifically, the prevalence of "high" scores changed for chronic sleep deprivation (9% to 43%, p = .0001). The prevalence of moderate depression increased from 4.3% to 29.8% (p = .0002). Only 4.3% reported a high level of burnout initially compared with 55.3% at year end (p < .0001). Scores that were originally more favorable than general population norms (p < .001) approached norms at the end of the year for empathic concern (p = .15). There was an association between becoming chronically sleep deprived and becoming depressed (OR = 7, p = .014). CONCLUSIONS: Given the association between chronic sleep deprivation and mood disturbances during internship, outcome assessment is warranted to see if duty-hour reform will translate into more hours slept or fewer hours worked, coincident with improved mood. PMID- 16377827 TI - Residents' responses to medical error: coping, learning, and change. AB - PURPOSE: To explore the significant emotional challenges facing resident physicians in the setting of medical mishaps, as well as their approaches to coping with these difficult experiences. METHOD: Twenty-six resident physicians were randomly selected from a single teaching hospital and participated in in depth qualitative interviews. Transcripts were analyzed iteratively and themes identified. RESULTS: Residents expressed intense emotional responses to error events. Poor patient outcomes and greater perceived personal responsibility were associated with more intense reactions and greater personal anguish. For the great majority of residents, their ability to cope with these events was dependent on a combination of reassurance and opportunities for learning. Interactions with medical colleagues and supervisory physicians were critical to this coping process. CONCLUSIONS: Medical mishaps have a profound impact on resident physicians by eliciting intense emotional responses. It is critical that resident training programs recognize the personal and professional significance of these experiences for young physicians. Moreover, resident education must support the development of constructive coping skills by facilitating candid discussion and learning subsequent to these events. PMID- 16377828 TI - Changing and sustaining medical students' knowledge, skills, and attitudes about patient safety and medical fallibility. AB - PURPOSE: To study the effects of a patient safety and medical fallibility curriculum on second-year medical students at the University of Missouri-Columbia School of Medicine in 2003-2004. METHOD: Students completed a knowledge, skills, and attitudes questionnaire before the curriculum, after the final learning experience, and one year later. A 95% confidence interval (CI) for paired differences assessed change over time. At one year, students also responded to items about their use of the curriculum, error reporting, and disclosure experiences. RESULTS: Fifty three of 92 students (55%) completed the questionnaire at all three assessment points. Students' eight items and the calculated knowledge score improved after the curriculum but only seven of these improvements were sustained one year. Responses to seven items did not change and five changed in an undesired direction after the curriculum and/or after one year. Seventy two students completed the self-reported behavior questions at one year. More than half reported using what they learned in the curriculum. Although 76% of students reported observing an error, 71% of these disclosed an error to their peers, 56% to a resident, and 46% to faculty. Only 7% reported an error using our electronic error reporting system. CONCLUSIONS: The curriculum led to changes in second-year medical students' knowledge, skills, and attitudes, but not all of the changes were sustained at one year, were in the desired direction, or were supported by their self-reported behaviors. The extent to which other informal or hidden curriculum experiences reversed the gains and affected the changes at one year is unknown. PMID- 16377829 TI - Myasthenic crisis. AB - Myasthenic crisis is a serious occurrence, affecting up to 27% of patients with myasthenia gravis. In the approach to the patient with myasthenic crisis, (1) the diagnosis of myasthenia gravis is confirmed; (2) respiratory failure is evaluated and treated in the intensive care unit, while potential precipitating factors are identified and managed; (3) immunomodulatory treatment is initiated; and (4) complications are avoided or managed promptly. The mortality rate should be 5% or less, with the elderly being most vulnerable. PMID- 16377831 TI - Risk of extubation failure in patients with myasthenic crisis. AB - INTRODUCTION: Weaning patients with myasthenic crisis (MC) from mechanical ventilation is often difficult, and the ideal time for extubation is often uncertain. However, little is known about the risk of extubation failure and the factors that may affect its occurrence. The goals of this study were to assess the risk of extubation failure in patients with MC and to determine which clinical variables may predict unsuccessful extubation. MATERIALS AND METHODS: Retrospective review of consecutive patients admitted for MC. Weaning method was categorized as T-piece or continuous positive airway pressure (i.e., T-piece trials not performed). Extubation failure was defined as need for reintubation or tracheostomy due to persistent neuromuscular insufficiency. Functional outcome was assessed using the modified Rankin score. RESULTS: We identified 26 episodes of MC in 20 patients. Median age was 42.1 years (range 14-83 years). Most patients were treated with immunomodulatory therapy (73%). There were seven episodes of extubation failure (prevalence rate 27%). Median time to reintubation was 36 hours. Older age (p = 0.05), atelectasis (p < 0.01), and pneumonia (p =0.02) were significantly associated with extubation failure. Patients with failed extubation had considerably prolonged intensive care unit stays (median 28 versus 7 days; p < 0.01) and hospital stays (median 40 versus 12 days; p < 0.01). CONCLUSION: Extubation failure may often complicate MC. Older age and development of pulmonary complications during mechanical ventilation increase the risk of extubation failure. PMID- 16377832 TI - Neurological recovery after decompressive craniectomy for massive ischemic stroke. AB - INTRODUCTION: Decompressive craniectomy has demonstrated efficacy in reducing morbidity and mortality in critically ill patients with massive hemispheric cerebral infarction. However, little is known about the patterns of functional recovery that exist in patients after decompressive craniectomy, and controversy still exists as to whether craniotomy and infarct resection ("strokectomy") are appropriate alternatives to decompression alone. We therefore used functional magnetic resonance imaging (f-MRI) to assess the extent and location of functional recovery in patients after decompressive craniectomy for massive ischemic stroke. METHODS: f-MRI was obtained in three patients with massive nondominant cerebral infarction who had undergone decompressive craniectomy for severe cerebral edema 13 to 26 months previously. Brain activation was triggered by hand-gripping or foot- movement tasks. Imaging results were combined with periodic clinical follow-up to determine the extent of neurological recovery. RESULTS: Activation of the contralateral hemisphere was seen in the sensorimotor cortex, premotor, and supplementary motor areas. Lesser activation patterns were seen in equivalent regions of the infarcted hemisphere. Peri-infarct activation foci were seen in two of the three patients, but no activation occurred within the area of infarction as defined by the initial stroke seen on diffusion weighted MRI. All three patients demonstrated some corresponding neurological improvement. CONCLUSION: After massive hemispheric cerebral infarction requiring decompressive craniectomy, patients may experience functional recovery as a result of activation in both the infarcted and contralateral hemispheres. The evidence of functional recovery in peri-infarct regions suggests that decompression alone may be preferable to strokectomy where the risk of damage to adjacent nonischemic brain may be greater. PMID- 16377833 TI - Endocrine alterations in critically ill patients with stroke during the early recovery period. AB - INTRODUCTION: Endocrine abnormalities in critically ill patients with stroke during the early recovery period have not been well characterized. METHODS: To investigate this issue, 33 consecutive mechanically ventilated patients (27 men) with hemorrhagic (n = 21) or ischemic (n = 12) stroke having a mean age of 57 +/- 12 years were studied. Glasgow Coma Scale score on admission in the hospital was 8 +/- 3. The following basal hormones were measured within 72 hours postextubation: cortisol, corticotropin (ACTH), free thyroxine, tri iodothyronine, thyroid-stimulating hormone, testosterone, estradiol, follicle stimulating hormone, luteinizing hormone, growth hormone, and insulin-like growth factor (IGF)-1. Subsequently, a low-dose (1 microg) ACTH stimulation test was performed. RESULTS: Twenty-six (79%) patients showed endocrine alterations. The most common change was low IGF-1 levels compatible with growth hormone deficiency (45%), followed by hypogonadism (39%), thyroid dysfunction (36%), and cortisol hyporesponsiveness (33%). CONCLUSION: Neuroendocrine changes occur with high frequency in critically ill patients with stroke during the early recovery period. It remains to be determined whether these changes have implications for functional and/or clinical outcome. PMID- 16377830 TI - Approach to neuromuscular disorders in the intensive care unit. AB - Neuromuscular disorders increasingly are recognized as a complication in patients in the intensive care unit (ICU) and represent a common cause of prolonged ventilator dependency. The distinct syndromes of critical illness myopathy, prolonged neuromuscular blockade, and critical illness polyneuropathy (CIP) may arise as a consequence of sepsis, multi-organ failure, and exposure to various medications--notably, intravenous corticosteroids and neuromuscular blocking agents--but the pathophysiology of these disorders remains poorly understood. More than one syndrome may occur simultaneously, and the distinctions may be difficult in a particular patient, but a specific diagnosis usually can be established after careful clinical, electrodiagnostic, and, when necessary, histological evaluation. For example, asthmatics requiring treatment with corticosteroids and neuromuscular blocking agents may develop an acute myopathy characterized by generalized weakness, preserved eye movements, elevated creatine kinase levels, and myopathic motor units on electromyography (EMG). Muscle biopsy demonstrates distinctive features of thick (myosin) filament loss on ultrastructural studies. Conversely, those with a prolonged ICU course that is complicated by episodes of sepsis with failure to wean from the ventilator, distal or generalized flaccid limb weakness, and areflexia probably have CIP. EMG in these patients demonstrates reduced or absent motor and sensory potentials with neurogenic motor units. Prolonged neuromuscular blockade most commonly occurs in patients with renal failure who have received prolonged infusions of neuromuscular blockers. There is severe flaccid, areflexic paralysis with normal sensation, facial weakness, and ophthalmoparesis that persists for days or weeks after the neuromuscular blockers have been discontinued. Repetitive nerve stimulation shows a decrement of the compound muscle action potential and, in most cases, establishes a disorder of neuromuscular transmission. With the recent epidemic of West Nile virus infection, a clinical syndrome of acute flaccid paralysis with several features indistinguishable from poliomyelitis has emerged. This article critically examines the clinical, electrophysiological, and pathological features of these and other acute neuromuscular syndromes that arise in the context of ICU care and summarizes the current understanding of the pathophysiology and treatment of these disorders. PMID- 16377834 TI - Ventricular cerebrospinal fluid hypocretin-1 inversely correlates with glucose levels in cerebrospinal fluid and serum from patients with neurological injuries. AB - INTRODUCTION: Hypocretin-1 is a hypothalamic neuropeptide that may help regulate arousal and feeding behavior and is quantifiable in cerebrospinal fluid (CSF). In this retrospective pilot study, hypocretin-1 levels obtained from ventricular CSF of neurologically injured patients were correlated with clinical and laboratory results to test whether arousal or metabolic factors might be related to the level of hypocretin-1. METHODS: CSF samples from a heterogeneous group of neurosurgical patients with externally draining intraventricular catheters were assayed in a standard manner for hypocretin-1 and other routine laboratories. Associations were sought between hypocretin-1 and clinical data such as body mass index (BMI), temperature, and Glasgow Coma Scale (GCS) score and between hypocretin-1 and laboratory data such as serum and CSF glucose, protein, and cell counts. RESULTS: Lower levels of ventricular CSF hypocretin-1 were correlated with higher levels of serum (p = 0.020) and ventricular CSF glucose (p = 0.001). Clinical findings such as BMI, temperature, and GCS failed to correlate with hypocretin-1. CONCLUSIONS: In a group of neurologically injured patients, hypocretin-1 and glucose levels are inversely correlated. More studies are needed to investigate these associations, particularly in a homogenous patient sample. PMID- 16377835 TI - Documentation in medical records improves after a neurointensivist's appointment. AB - INTRODUCTION: Medical documentation is important for communication among health care professionals, research, legal defense, and reimbursement. Previous studies have indicated insufficient documentation by health care providers and resistance among physicians to comply with the new guidelines. Data in the intensive care unit (ICU) subpopulation are scarce. We examined the hypothesis that a newly appointed neurointensivist may alter documentation practices in a university hospital setting. METHODS: We sampled medical records of neurological intensive care unit (NICU) patients admitted with three specific diagnoses (head trauma, intracerebral hemorrhage, and subarachnoid hemorrhage) and examined changes in the documentation of important prognostic variables in two time periods: before and after the appointment of a neurointensivist. RESULTS: Overall, documentation improved from 32.5 to 57.5% (odds ratio, 95% confidence interval 2.8, 1.9-4.2) in the after period. Documentation using Glasgow Coma Scale, clot volume, Hunt & Hess scale, and Fisher's grade also improved significantly in each of the diagnoses examined in the after period. CONCLUSIONS: Our findings suggest that a major change was implemented in the NICU regarding documentation after a neurointensivist was appointed. Although the direct or indirect impact of the appointment was not clarified, these preliminary data warrant a prospective ICU study, which should determine the exact variables that play a role in documentation, how they change over time, and what reinforcing mechanisms can be used. PMID- 16377836 TI - Frameless stereotactic aspiration and thrombolysis of spontaneous intracerebral hemorrhage. AB - INTRODUCTION: To test the feasibility and safety of a minimally invasive technique, we report our experience in treating spontaneous intracerebral hemorrhage (ICH) patients by using frameless stereotactic clot aspiration thrombolysis and its effects on their 30-day survival. We compared the observed cohort mortality with its predicted 30-day ICH mortality, by using previously validated methods. METHODS: Selection criteria were diagnosis of hypertensive ICH > or =35 cc, reduced level of consciousness, and no brainstem compression. Frameless stereotactic puncture/clot aspiration followed by intraclot external catheter placement was performed. Two milligrams of recombinant tissue plasminogen activator (rtPA) was administered q12 hours until ICH volume < or =10 cc, or the catheter fenestrations were no longer in continuity with the clot. RESULTS: Fifteen patients were treated, mean age was 60.7 years. Hemorrhage locations included basal ganglia (13), thalamic (1), and lobar (1); mean systolic blood pressure; and admission ICH volumes were 229.3 mmHg and 59.1 cc, respectively. Median time from ictus to clot aspiration/thrombolysis was 1 (range 0-3) day. Mean hematoma volume was reduced to 17% of pretreatment size. Complications were ventriculitis (6.6%) and clot enlargement (13.3%). Two patients were dead at 30 days. Median Glasgow Coma Scale (GCS) scores were 10.5 (4-15) at admission and 11.0 (3-15) at discharge. By using the most conservative estimate for analysis, probability of observing two or fewer deaths among 15 patients with an overall probability of dying calculated at 0.33 was p = 0.079. CONCLUSIONS: In this selected cohort of patients with ICH, stereotactic aspiration and thrombolytic washout seemed to be feasible and to have a trend towards improved 30-day survival, when using their predicted mortality data as "historical control." Complications did not exceed expected incidence rates. Based on the experience presented here as well as previous similar reports, a larger, randomized study addressing dose escalation, patient selection, and best therapeutic window is needed. PMID- 16377837 TI - Systemic hemostasis with recombinant-activated factor VII followed by local thrombolysis with recombinant tissue plasminogen activator in intraventricular hemorrhage. AB - INTRODUCTION: A 51-year-old woman on warfarin thromboprophylaxis for transient ischemic attacks developed sudden onset nausea, vomiting, and decreased mental status, rapidly becoming comatose. Head computed tomography (CT) showed intracerebral hemorrhage, extending into all ventricular chambers, and acute obstructive hematocephalus requiring urgent ventricular drainage. CT angiogram showed no evidence of an aneurysm or vascular malformation. METHODS: The pretreatment international normalized ratio (INR) of 4.9 was rapidly corrected with recombinant activated factor VII and an external ventricular drain was placed. Despite accurate positioning, the ventriculostomy thrombosed and became nonfunctional. Recombinant tissue plasminogen activator was given intraventricularly and resulted in partial ventricular decompression within 24 hours, with dramatic improvement in the patient's level of consciousness. RESULTS: Repeated intraventricular fibrinolysis resulted in further reduction of the intraventricular hematoma within a few days and a good patient outcome. The patient did not require permanent ventricular shunt. CONCLUSION: To our knowledge, this is the first reported case of combined systemic enhancement of hemostasis and local fibrinolysis as a life-saving measure in intracranial hemorrhage. PMID- 16377838 TI - Fatal pulmonary embolism despite an inferior vena cava filter in glioblastoma multiforme. AB - INTRODUCTION: Patients with high-grade gliomas are at increased risk of pulmonary embolism and deep vein thrombosis. Treatment of this complication remains uncertain and options include anticoagulation or venous filters. METHODS: We present a patient with a glioblastoma multiforme who developed a fatal pulmonary embolism in spite of an inferior vena cava filter. CONCLUSION: In spite of the theoretical risk of intratumoral hemorrhage, anticoagulation may be more effective and associated with fewer complications than inferior vena cava filters. PMID- 16377839 TI - Magnetic resonance restricted diffusion resolution correlates with clinical improvement and response to treatment in herpes simplex encephalitis. AB - INTRODUCTION: A 34-year-old man presented with herpes simplex encephalitis (HSE), with magnetic resonance imaging (MRI) showing dense foci of restricted diffusion in the temporal lobe. CASE REPORT: With treatment and clinical improvement, follow-up MRI done 8 days later showed complete resolution of the restricted diffusion abnormalities, whereas other MRI sequences suggested interval progression. DISCUSSION: Restricted diffusion abnormalities on MRI in patients with HSE may be more sensitive to and correlate better with disease activity in HSE. PMID- 16377840 TI - Minor trauma may lead to traumatic internal carotid artery dissection. AB - A farmer's wife was slapped by her cow's tail during milking. There was no obvious injury, but afterward she first developed unusual headaches of unknown origin and then a severe stroke 7 days later. Further diagnostics, including magnetic resonance imaging techniques, revealed a dissection of the carotid artery to be the reason for the stroke, probably caused by being hit by the cow's tail. This case demonstrates that persisting unusual headaches, even after minor trauma, should lead to further diagnostic procedures and then therapeutic intervention for stroke prevention if necessary. PMID- 16377841 TI - The syndrome of irreversible acidosis after prolonged propofol infusion. AB - INTRODUCTION: Propofol infusion syndrome is described in the pediatric literature as metabolic acidosis, rhabdomyolysis, and bradycardia that results in death. The pathogenesis of this syndrome is thought to be activation of the systemic inflammatory response, which culminates in acidosis and muscle necrosis. MATERIALS AND METHODS: Retrospective chart review of three patients in the Neurological Critical Care Units at Hahnemann and Massachusetts General Hospitals between October 2001 and September 2004. RESULTS: Patient 1: A 27-year-old woman had seizures secondary to hemorrhage from an arteriovenous malformation. Propofol coma was induced for sedation. After initiation of propofol, she developed a metabolic acidosis, hypotension, and bradycardia and expired. Patient 2: A 64 year-old man presented in status epilepticus. After prolonged propofol administration, he developed metabolic acidosis, hypotension, and rhabdomyolysis and expired. Patient 3: A 24-year-old woman presented in status epilepticus secondary to encephalitis. Propofol was added for seizure control. She developed hypotension, metabolic acidosis, and bradyarrhythmias. Despite transvenous pacing, she expired. CONCLUSION: These data show an association between extended propofol use and metabolic acidosis, rhabdomyolysis, and death in adults, as well as children. Risk factors for propofol infusion syndrome in adults include lean body mass index, high dose, and administration of more than 24-hour duration. Creatine phosphokinase, lactic acid levels, electrolytes, and arterial blood gases should be monitored frequently. Both bacterial and fungal cultures should be obtained. If this syndrome is suspected, hemodialysis should be considered. In fatal cases, autopsy should include electron microscopy of cardiac and skeletal muscle to look for mitochondrial dysfunction. Further study is warranted. PMID- 16377842 TI - Brain tissue oxygen monitoring in intracerebral hemorrhage. AB - INTRODUCTION: Brain tissue oxygen (PbrO2) monitoring is an emerging technique for detection of secondary brain injury in neurocritical care. Although it has been extensively reported in traumatic brain injury and aneurysmal subarachnoid hemorrhage, its use in nontraumatic intracerebral hemorrhage (ICH) has not been well described. We report complementary preliminary studies in a large animal model and in patients that demonstrate the feasibility of PbrO2 monitoring after ICH. METHODS: To assess early events after ICH, Licox Clark-type oxygen probes were inserted in the bilateral frontal white matter of four anesthetized swine that subsequently underwent right parietal hematoma formation in an experimental model of ICH. Intracranial pressure (ICP) was monitored as well. Seven patients with acute ICH, who were undergoing ICP monitoring as part of standard neurocritical care, had placement of a frontal oxygen probe, with subsequent monitoring for up to 7 days. RESULTS: In the swine ICH model, a rise in ICP early after hematoma formation was accompanied by a decrease in ipsilateral and contralateral PbrO2. Secondary increases in hematoma volume resulted in further decreases in PbrO2 over the first hour after ICH. In patients undergoing oxygen monitoring, low PbrO2 (<15 mmHg) was common. In these patients, changes in FiO2, mean arterial pressure, and cerebral perfusion pressure (but not ICP) predicted subsequent change in PbrO2. CONCLUSION: Brain tissue oxygen monitoring is feasible in ICH patients, as well as in a swine model of ICH. Translational research that emphasizes complementary information derived from human and animal studies may yield additional insights not available from either alone. PMID- 16377845 TI - Report from the Indian Third National Workshop on Critical Care Neurology. PMID- 16377843 TI - Causes of Cheyne-Stokes respiration. AB - Cheyne-Stokes respiration (CSR) is one of several types of unusual breathing with recurrent apneas (dysrhythmias). Reported initially in patients with heart failure or stroke, it was then recognized both in other diseases and as a component of the sleep apnea syndrome. CSR is potentiated and perpetuated by changing states of arousal that occur during sleep. The recurrent hypoxia and surges of sympathetic activity that often occur during the apneas may have serious health consequences. Heart failure and stroke are risk factors for sleep apnea. The recurrent apneas and intermittent hypoxia occurring with sleep apnea further damage the heart and brain. Although all breathing dysrhythmias do not have the same cause, instability in the feedback control involved in the chemical regulation of breathing is the leading cause of CSR. Mathematical models have helped greatly in the understanding of the causes of recurrent apneas. PMID- 16377846 TI - Purification and characterization of Bacillus cereus protease suitable for detergent industry. AB - An extracellular alkaline protease from an alkalophilic bacterium, Bacillus cereus, was produced in a large amount by the method of extractive fermentation. The protease is thermostable, pH tolerant, and compatible with commercial laundry detergents. The protease purified and characterized in this study was found to be superior to endogenous protease already present in commercial laundry detergents. The enzyme was purified to homogeneity by ammonium sulfate precipitation, concentration by ultrafiltration, anion-exchange chromatography, and gel filtration. The purified enzyme had a specific activity of 3256.05 U/mg and was found to be a monomeric protein with a molecular mass of 28 and 31 kDa, as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and nondenaturing PAGE, respectively. Its maximum protease activity against casein was found to be at pH 10.5 and 50 degrees C. Proteolytic activity of the enzyme was detected by casein and gelatin zymography, which gave a very clear protease activity zone on gel that corresponded to the band obtained on SDS-PAGE and nondenaturing PAGE with a molecular mass of nearly 31 kDa. The purified enzyme was analyzed through matrix-assisted laser desorption ionization-time-of flight-mass spectrometry (MALDI-TOF-MS) and identified as a subtilisin class of protease. Specific serine protease inhibitors, suggesting the presence of serine residues at the active site, inhibited the enzyme significantly. PMID- 16377847 TI - Preparation of trehalase inhibitor validoxylamine A by biocatalyzed hydrolysis of validamycin A with honeybee (Apis cerana Fabr.) beta-glucosidase. AB - Validoxylamine A is structurally similar to trehalose and acts a potent competitive inhibitor of trehalase. It has recently been receiving increased attention as a potential material for the development of new insecticides or drugs. In this study, beta-glucosidase extracted from honeybees (Apis cerana Fabr.) was used as a catalyst to produce validoxylamine A through enzymatic hydrolysis of validamycin A. Beta-glucosidase was separated and purified from honeybees, and its characteristics were examined. The results showed that beta glucosidase was stable across a range of temperatures from 30 to 40 degrees C and across a relatively wide range of pH values from 5.0 to 7.5. Investigation of the biocatalyzed hydrolysis process from validamycin A to validoxylamine A with beta glucosidase revealed that both the substrate (validamycin A) and the product (validoxylamine A) inhibited beta-glucosidase activity. The inhibition constant of the substrate Kis value was 5.01 mM, and that of the product Kip value was 1.32 mM. This product inhibition was competitive. PMID- 16377848 TI - Reactive oxygen species, cell growth, and taxol production of Taxus cuspidata cells immobilized on polyurethane foam. AB - Dynamic changes in reactive oxygen species (ROS) of Taxus cuspidata cells immobilized on polyurethane foam were investigated and the relation between ROS content and taxol production was discussed. Immobilization shortened the lag period of cell growth and moderately increased H2O2 and O2-* contents inside the microenvironment within the first 15 d. After 20 d, excessive production of H2O2 and O2-* was observed accompanied by marked increases in membrane lipid peroxidation and cell membrane permeability. The taxol content of immobilized cells was fourfold that of suspended cells at d 35. The addition of exogenous H2O2 barely affected malondialdehyde content and cell membrane permeability but led to an obvious accumulation of taxol. It is inferred that the intracellular and extracellular H2O2 inside the microenvironment might be one factor promoting taxol biosynthesis under the immobilization stress. PMID- 16377849 TI - Uncoupled metabolism stimulated by chemical uncoupler and oxic-settling-anaerobic combined process to reduce excess sludge production. AB - The effects of three uncoupled metabolic systems (conventional activated sludge process with the addition of 3,3', 4',5-tetrachlorosalicylanilide [TCS], oxic settling-anaerobic [OSA] process modified by insertion of a sludge-holding tank in the sludge return line, and TCS and OSA combined process) on reducing excess sludge production were studied. Compared with the control conventional activated sludge process, the most effective system was the combined process, which could reduce excess sludge production by 46.90%. The 180-d operation results confirmed that TCS is an effective chemical uncoupler in reducing the sludge yield but that it had an adverse effect on substrate removal capability, effluent nitrogen concentration, and sludge settleability. The OSA process decreased excess sludge production by only 26% but had less adverse effect on effluent quality and could improve sludge settleability. The effluent total phosphorous concentration of the three systems was slightly lower than of the control unit. Microbial populations were monitored by both microscopic and molecular biologic analysis method (polymerase chain reaction [PCR]-denaturing gradient gel electrophoresis [DGGE]). The presence of TCS caused metazoans to disappear and decreased the number and activity of protozoa. PCR amplification of 16S rRNA and sequent DGGE analysis found a shift in the diversity of the predominant species. The results imply that OSA combined with the chemical uncoupler process may effectively reduce excess sludge yield and not affect process performance significantly. PMID- 16377850 TI - A new thermostable peroxidase from garlic Allium sativum: purification, biochemical properties, immobilization, and use in H2O2 detection in milk. AB - Analysis of peroxidase activity by native polyacrylamide gel electrophoresis (PAGE) from a garlic bulb (Allium sativum L) extract showed two major activities (designated POX1 and POX2). The POX2 isoenzyme was purified to homogeneity by ammonium sulfate precipitation, gel filtration, and cation-exchange chromatography. The purified enzyme was found to be monomeric with a molecular mass of 36.5 kDa, as determined by sodium dodecyl sulfate-PAGE. The optimum temperature ranged from 25 to 40 degrees C and optimum pH was about 5.0. The apparent Km values for guaiacol and H2O2 were 9.5 and 2 mM, respectively. POX2 appeared highly stable since 50% of its activity was conserved at 50 degrees C for 5 h. Moreover POX2 was stable over a pH range of 3.5-11.0. Immobilization of POX2 was achieved by covalent binding of the enzyme to an epoxy-Sepharose matrix. The immobilized enzyme showed great stability toward heat and storage when compared with soluble enzyme. These properties permit the use of this enzyme as a biosensor to detect H2O2 in some food components such as milk or its derivatives. PMID- 16377851 TI - Antiretroviral drug resistance and resistance testing. AB - Antiretroviral resistance testing should be performed in newly diagnosed patients with acute or recent HIV infection and at the time of treatment failure, and there is growing support for testing in newly diagnosed, treatment-naive patients with chronic infection as well. Genotypic testing is preferred for baseline screening, because it is more sensitive than phenotypic testing for the presence of mixed populations of drug-susceptible and -resistant virus and because it is less expensive. Phenotypic testing provides quantitative information on the degree of resistance and is also able to assess interactions among mutations. As a result, it can be particularly useful in determining treatment options for treatment-experienced patients with multi-drug resistant virus. In many cases, there may be advantages to the use of both tests. PMID- 16377852 TI - Oral manifestations of HIV disease. AB - HIV-related oral conditions occur in a large proportion of patients, and frequently are misdiagnosed or inadequately treated. Dental expertise is necessary for appropriate management of oral manifestations of HIV infection or AIDS, but many patients do not receive adequate dental care. Common or notable HIV-related oral conditions include xerostomia, candidiasis, oral hairy leukoplakia, periodontal diseases such as linear gingival erythema and necrotizing ulcerative periodontitis, Kaposi's sarcoma, human papilloma virus associated warts, and ulcerative conditions including herpes simplex virus lesions, recurrent aphthous ulcers, and neutropenic ulcers. PMID- 16377853 TI - Dermatologic manifestations of HIV infection. AB - Although some dermatologic diseases have decreased markedly in frequency in the potent antiretroviral therapy era, other conditions remain common. Among patients with low CD4(+) cell counts who are not on or not adherent to antiretroviral therapy, notable conditions include psoriasis, photodermatitis, prurigo nodularis, molluscum, and adverse drug reactions. Conditions that remain relatively common despite adequate antiretroviral therapy include eczema, xerosis, warts, and Kaposi's sarcoma. Disorders that are associated with immune reconstitution under potent antiretroviral therapy include acne, staphylococcal infections, and erythema nodosum. In addition, HIV and hepatitis C virus (HCV) coinfection is associated with a number of skin disorders. PMID- 16377854 TI - Insulin Autoimmune Syndrome (Hirata's disease): severe hypoglycemic episodes in Graves' hyperthyroidism patient treated with methimazole. PMID- 16377855 TI - International symposium on infectious agent transmission model building--focusing on assessment of risk to communities. AB - The Susceptible Infected and Recovery (SIR) Model proposed by Robert May in the UK is the basis of the present mathematical model building of infectious disease epidemics. Need for model building incorporating more social and other relevant factors has been recognized. An important example is the introduction of idea of the scale-free distribution of links among the people. More refined models by taking into account the nature of a pathogen, geo-sociological factors, lifestyles of the people, etc., have been developed. For example, Koopman proposed a model for prediction of epidemic expansion based on actual epidemiological data. Eubank proposed a model assessing the bio-terror attack using a model city where every day activity is going on. The present workshop, participated by experts from the US, the UK and Japan, is the first meeting of the proposed series of conference on this issue. PMID- 16377856 TI - Modeling on social spread from immunity. AB - We are now planning to make a transmission model of infectious diseases in the scale of a city. People live in the city contacting other persons with daily life. The model regards a contact as a source of infection. A person will be simulated as a simple system of differential equations. As a candidate of differential equations, we are now investigating Marchuk's simple model. We adopt Marchuk's simple model because it has formation time, i.e., latent time. As Dr. Takeuchi showed, latent time is very important. There remain problems of choosing parameters for special diseases. We are now planning to use Marquardt method to minimize residuals form clinical data to estimate parameters. As for contacts, there are many approaches. The approach of the MIDAS project is very intensive. Our approach is simple. There are about 30,000 Japanese every 15 minutes daily life data, sleeping, eating, work, study, house keeping, etc. Our approach is to make virtual families, husband, wife, children in a city and assign actions from the every 15 minutes data statistically and estimate their contacts in the companies or schools, etc. PMID- 16377857 TI - Effectiveness of vaccination strategies for infectious diseases according to human contact networks. AB - Mathematical modeling of infectious disease transmission aims to predict the spread of infection and the effectiveness of containment strategies. For the case of infectious diseases that transmit between human, one key factor becomes the modeling of contacts between individuals through which the disease is potentially transmitted. We review several formulations of such contacts with emphasis on the recently investigated scale-free network, and also discuss the effectiveness of vaccination strategies. PMID- 16377858 TI - The hospital with more beds has a higher probability of experiencing an outbreak of the hospital infection. AB - The dependence of the incidence of the hospital infection on the number of the beds was studied by using SIR model with the computer simulation. We obtained the result that the hospital with more beds has a higher probability of experiencing an outbreak. The integration of smaller hospitals into a larger hospital was once considered economically advantageous and such a policy has been implemented in Japan since the early 1980s. This analysis, however, warns against the policy. PMID- 16377859 TI - HIV transmission webs: HIV infection trends in Japan in 1989-2004. AB - HIV infection is exponentially increasing in the past decade among young homosexuals in Japan. Possible influence of social webs among young males whose creation was facilitated by electronic innovation is discussed. PMID- 16377860 TI - Infection transmission science and models. AB - Infection transmission systems circulate infection through complex contact patterns related to both contact patterns and patterns of factors that affect the risk of transmission given contact. The nonlinear dynamics of infection transmission cause these patterns to make big differences in population infection levels. A science of infection transmission system analysis is needed to focus on those details that affect the control of infection transmission. This science must have a strong theoretical base because there is little chance that a dominantly data based approach not using mechanistic models of transmission will have any predictive value. The theoretical base should be built on linked transmission system models that are focused on making needed inferences for both building the theoretical base and making infection control decisions. The linking of different models is needed for a strategy of inference robustness assessment that is designed to find the model that is simple enough to effectively analyze the transmission system but not so simple that realistic violation of simplifying assumptions will change an inference. Types of models that should be used in such linked analyses include deterministic and stochastic compartmental models, discrete individual models with individual event histories but structured mass action mixing, network models that provide more detail as to who has contact with whom, and intermediate model forms such as correlation models that address some aspects of contact details while preserving the flexibility of deterministic compartmental models to structure mixing and analyze the system. While transmission system science is currently weak in regards both to its data base and its theory base, many things are now coming together that could make this science flourish. On the data side these include greater ability to detect infectious agent sequences in the environment and greater ability to sequence and genetically relate agents identified at different sites in the transmission system. On the theory sides, new model construction and model analysis methods are providing new potential to use the new sources of data. Also new parameter estimation methods provide new potential to combine models and data in effective analytic strategies. PMID- 16377861 TI - Network based models of infectious disease spread. AB - It has recently become possible to simulate directly dynamics on very large networks. This paper describes a model of epidemiology on a social network, the Epidemiological Simulation System (EpiSims), and offers general speculation on analyzing disease dynamics on networks. We describe the process of building a realistic social network, describe several different definitions of the network, each useful for certain purposes. Finally, we raise some important questions about structural properties of networks and how they influence dynamics. PMID- 16377862 TI - Shigella dysenteriae type 1-induced diarrhea in rats. AB - With the aim of setting up an animal model of Shigella dysenteriae-induced diarrhea, Wistar rats received per os increasing densities of S. dysenteriae type 1 (Sd1). Inoculum of 12 x 10(8) Sd1 provoked dysenteric diarrhea within 24 h. Feces of healthy rats were molded, brown to black and rough. Rats developing diarrhea presented blood at the anal orifice; stools were soft or liquid containing mucus, or molded, smooth and mucus-coated. At times, stools appeared longer, dark and shiny due to the presence of mucus and blood, or molded, lumpy and brittle. Diarrheal induction was associated with abdominal ailment, progressive increase in stool weight and frequency, and increase in bacterial population. Sixty-seven percent of the total number of deaths had occurred by day 6 after diarrheal induction. These results indicate that Sd1 induced in rats a model of shigellosis which might be helpful for physiopathological and pharmacological studies of this type of infectious diarrhea. PMID- 16377863 TI - Changes in antibiotic use, cost and consumption after an antibiotic restriction policy applied by infectious disease specialists. AB - The study was designed to compare antibiotic use, cost and consumption before and after an initiation of an antibiotic-restriction policy in our hospital. The policy was applied in 2003, and the prescription of two groups of antibiotics (intravenously used and expensive antibiotics) was restricted. A prescription for the restricted antibiotics could be obtained with approval by an infectious disease specialist (IDS). All the hospitalized patients who received antibiotics were evaluated by a cross-sectional study with standard criteria. The annual cost and consumption of antibiotics were evaluated. After restriction, the rate of antibiotic use decreased from 52.7 to 36.7% (P < 0.001), and the appropriate use increased from 55.5 to 66.4% (P < 0.05). Appropriate use was higher for restricted antibiotics (88.4%) than for unrestricted ones (58.2%) (P < 0.001), and higher in the presence of ID consultation (97.5%) than in the absence of consultation (55.7%) (P < 0.001). Culture-based treatment was increased, and appropriate use in such cases (93.0%) was higher than empirical treatment (33.3%) (P < 0.001). After the restriction policy, consumption of antibiotics belonging to the restricted groups was decreased by 44.8%. Total expenditure of all antibiotics was decreased by 18.5%, and the savings were US$332,000 per year. This restriction policy was effective in promoting rational antibiotic prescription and lowering antibiotic cost and consumption in our hospital. PMID- 16377864 TI - Emergence of non-albicans Candida species and antifungal resistance in a tertiary care hospital. AB - The spectrum of candidiasis has changed with the emergence of non-albicans Candida spp. and acquired antifungal resistance, especially in immunocompromised hosts. This changing scenario has necessitated routine antifungal susceptibility testing. In the present work, 102 Candida spp. isolates gathered during 2003 - 2004 were characterized by standard procedures, and antifungal susceptibility testing to amphotericin B, fluconazole and itraconazole was performed by broth macrodilution (BMD)-minimum inhibitory concentration (MIC) and disk diffusion (DD) methods. Among all isolates, 77.4% were from an ICU and 10.8% were obtained from a nursery. The majority of the isolates were C. tropicalis (48%), followed by C. parapsilosis (27.4%) and C. albicans (22.5%). Overall 6.9, 4.9 and 3.9% of all isolates were resistant to amphotericin B, fluconazole and itraconazole, respectively. Out of the 5 (4.9%) isolates resistant to fluconazole, 4 (3.9%) were from patients with AIDS on fluconazole prophylaxis. A discrepancy was observed between the results of susceptibility testing by DD and those by BMD MIC: 15 (14.7%) isolates were reported to be resistant by DD despite having low MICs. Based on these results, it was concluded that initial antifungal screening of clinical isolates by the DD method followed by confirmation of resistant strains by the broth dilution method is desirable to optimize patient management. PMID- 16377865 TI - SEN virus prevalence among non-B and non-C hepatitis patients with high liver function tests in the south of Turkey. AB - We investigated the characteristics and detection rates of SEN virus (SENV) infection among 100 Turkish patients who had with high alanine aminotransferase (ALT) and aspartate aminotransferase levels but were negative for HBV DNA and HCV RNA and had no history of transfusion. As a control group, we also analyzed 50 healthy individuals who had normal ALT levels, were negative for HBV DNA and HCV RNA, and had no history of transfusion. The serum samples of patient and controls were analyzed by PCR to detect the presence of SENV DNA and its two genotypes (SENV-H and SENV-D). We detected SENV DNA in 13 of 100 (13%) patients. Five of 13 (38.46%) patients were positive for SENV-D and 8 of 13 (61.53%) patients were positive for SENV-H DNA. We also detected SENV DNA in 5 of 50 (10%) patients in the control group. Two of 5 (40%) patients were positive for SENV-D and 3 of 5 (60%) patients were positive for SENV-H DNA in the control group. SENV was detected at almost the same frequency in the patient and control group. SENV did not seem to contribute to the pathogenesis of liver disease (P > 0.05) in this cohort. Our results also showed that SENV transmission was not only associated with blood transfusion but also with some other possible routes. PMID- 16377866 TI - Adverse effect of staphylococci slime on in vitro activity of glycopeptides. AB - Adhesion to biomaterial is assumed to be a crucial step in the development of staphylococcal foreign body infections. Production of extracellular slime has major implications for the development and implementation of therapeutic strategies. The effect of extracted slime was investigated on the activity of vancomycin, teicoplanin, linezolid, quinupristin/dalfopristin, rifampicin and ranbezolid against 10 clinical and 4 ATCC staphylococcal isolates. The slime extract caused a 2- to 16-fold increase in the MICs of vancomycin and teicoplanin, with a shift in the MIC(90) from 2 to 32 (vancomycin) and 2 to 16 (teicoplanin), whereas the MICs of linezolid and quinupristin/dalfopristin were only moderately affected. In time-kill studies, a significant decrease in bacterial killing (>3 log(10) cfu/ml) was observed with vancomycin and teicoplanin (4 x MIC) after addition of slime (5 and 20 mg/ml), whereas the effect of killing by linezolid and quinupristin/dalfopristin was very modest. The rifampicin and ranbezolid MICs and kill curves were not influenced by the addition of slime. The present study thus indicated that slime interferes with the antimicrobial effect of glycopeptide drugs (vancomycin, teicoplanin), and that for effective prevention and treatment of prosthetic device-related infections, appropriate and newer antibiotics such as ranbezolid should be considered. PMID- 16377867 TI - Rapid and quantitative detection of Crimean-Congo hemorrhagic fever virus by one step real-time reverse transcriptase-PCR. AB - In this article, the development of a new TaqMan-based one-step real-time reverse transcriptase-polymerase chain reaction (RT-PCR) assay for detection and quantification of Crimean-Congo hemorrhagic fever virus (CCHFV) RNA is described. Selected oligos targeting the highly conserved S region of CCHFV were designed by using our oligo design and analysis software, Oligoware 1.0. None of the primer sequences showed genomic cross-reactivity with other viruses or cells in a BLAST (NCBI) search analysis. The sensitivity and specificity of the primers and the probe were tested using 18 serum samples from patients from East Anatolian who were suspected of having CCHFV, including 2 samples that had already been confirmed to be positive for CCHFV. Among the 16 previously unconfirmed samples, 5 were positive by TaqMan-based one-step real-time RT-PCR and 1 was positive by non-nested RT-PCR, and these results were confirmed with DNA sequencing analysis. The 2 previously confirmed CCHFV RNA samples were also positive by both TaqMan based one-step real-time RT-PCR and non-nested RT-PCR tests. To ensure the quantitative reproducibility of TaqMan-based one-step real-time RT-PCR, the procedure was repeated several times and the same results were obtained (SD = 0.84 [maximum value]). The developed assay was able to sensitively quantify the concentration of CCHFV RNA, which ranged from 10(2) to 10(7) copies/ml per reaction, using plasmid standards generated from the CCHFV RNA (correlation coefficiency = 0.989). The results of the one-step real-time RT-PCR assay were more sensitive than those of the non-nested RT-PCR assay. It can be concluded that our one-step real-time RT-PCR assay is a reliable, reproducible, specific, sensitive and simple tool for the detection and quantification of CCHFV. PMID- 16377868 TI - Prevalence of Salmonella enterica serovar typhi in bile and stool of patients with biliary diseases and those requiring biliary drainage for other purposes. AB - Acute suppurative cholangitis is a serious complication of biliary obstructions due to infection. Salmonella enterica serovar Typhi is an important biliary pathogen. Bile samples from 445 patients with biliary diseases as well as those requiring biliary drainage for other miscellaneous gastrointestinal diseases were investigated bacteriologically with special emphasis on Salmonella. Fecal samples or rectal swabs were also obtained from 402 of these patients. Bactericholia was detected in 68.8% patients and Salmonella in 5.8% of all bile samples. Other strains of salmonellae were also present in a fair number of the samples. Some of the patients had the same type of bacterial isolates from their stool samples as those from their bile samples. Colonization of the biliary system may not be clinically apparent, but is associated with an increased risk of infection. PMID- 16377869 TI - Pyogenic liver abscess: a retrospective analysis of 107 patients during a 3-year period. AB - Pyogenic liver abscess (PLA) is a potentially life-threatening disease, and early diagnosis may be difficult. In order to provide diagnostic clues and to enhance the prompt management of such cases, we retrospectively investigated the clinical characteristics of PLA during a 3-year period in a tertiary-care hospital. The crude incidence rate of PLA in our study was 446.1 per 100,000 hospital admissions. Male predominance and a mean age of 57.6 +/- 14.4 years were observed. Diabetes mellitus was the most common concomitant disease, and biliary pathologies were the most common predisposing cause of this type of abscess. The most common clinical features were fever, chills, and abdominal pain. Leukocytosis was found in 67.3% of the patients, and the observed C-reactive protein (CRP) values were high. The most common pathogen was Klebsiella pneumoniae. The mortality rate was 6.5%. A complete history, physical examination, evaluation of the white blood cell count and CRP, and the prompt arrangement of imaging studies may lead to an earlier diagnosis. The aggressive performance of image-guided catheter drainage and the appropriate administration of antibiotics may reduce the mortality rate of PLA. PMID- 16377870 TI - Correlation between the usage volume of veterinary therapeutic antimicrobials and resistance in Escherichia coli isolated from the feces of food-producing animals in Japan. AB - We compared the overall usage of veterinary therapeutic antimicrobials in Japan to the proportion of antimicrobial-resistant. Escherichia coli isolated from the feces of apparently healthy food-producing animals in 2001. The annual sales volume of veterinary antimicrobials, which is published and accumulated information, was subdivided according to the target animal species (cattle, pigs, broiler chickens, and layer chickens). The antimicrobial susceptibility of the isolates was examined against 7 classes of 11 antimicrobials. The rates of antimicrobial resistance among the isolates were found to correlate significantly with the usage of antimicrobial agents in cattle, pigs, and broiler and layer chickens. Therefore, the overall usage of veterinary antimicrobials appears to contribute to the appearance of antimicrobial resistance in E. coli isolates from apparently healthy food-producing animals. PMID- 16377871 TI - Delayed diagnosis of tuberculous arthritis. AB - Monoarticular tuberculosis (TB) affecting the knee is rare in all forms of TB (0.1-0.3%). We present the case of a patient with tuberculous arthritis in whom the diagnosis was belated due to a lack of familiarity with the disease; here, we emphasize the difficulties associated with the diagnosing joint TB. A 20-year-old man was referred to our department due to swelling of the right knee and the presence of persistent, mild pain for 4 years. The lack of systemic evidence of this disease, the indolent course of disease, and the presence of non-specific symptoms renders early recognition of this disease difficult. Furthermore, in cases in which a diagnosis cannot be reached simply by culturing the synovial fluid, synovial biopsy cultures should be considered in the diagnostic process, due to the high rate of positivity of such cultures. The diagnosis and treatment of articular TB are both urgent matters; surgical debridement and strict adherence to antituberculous chemotherapy tend to yield a satisfactory functional outcome. PMID- 16377872 TI - A case of tuberculous pyomyositis that caused a recurrent soft tissue lesion localized at the forearm. AB - We present the case of a 20-year-old male who had a non-traumatic soft tissue lesion (4 x 3 cm) with recurrent discharge at his right posteromedial antebrachial muscles; the patient underwent surgery twice, and antibiotic therapy was administered, but no cure was achieved with these treatments. The patient underwent surgery at our medical center. There was no history of pulmonary, gastrointestinal, or genitourinary tuberculosis (TB). Due to suspected pulmonary, genitourinary, and gastrointestinal TB, radiography and computed tomography scans were performed, and these studies disclosed no evidence of a primary origin. The erythrocyte sedimentation rate and the results of purified protein derivate testing were normal. We also detected submandibular lymphadenopathy (LAP) (2 x 3 cm) localized at a submandibular site in our patient 4 months after his first visit to our clinic. Smears were stained with Ehrlich Ziehl Neelsen (EZN) stain and culture were grown for Mycobacterium tuberculosis complex (MTC); the samples used for these assays had been obtained by incisional biopsy of the forearm lesion and by aspiration of the submandibular lymph node, and they were found to be MTC-positive. Then, a culture for MTC, derived from an induced sputum sample, was found to be positive, despite the negative results obtained with a sputum smear subjected to EZN staining. According to these results, the primary focus of the tuberculous pyomyositis and the submandibular LAP was the lungs. The lesion and submandibular LAP were both treated successfully by the administration of antituberculous chemotherapy. PMID- 16377873 TI - Interruption of env gene expression depending on the length of the SV40 early region used for the polyA signal. AB - In order to invent a screening system to check in vivo gene function and the efficiency of gene transfer mediated by a retroviral vector system, we established a novel packaging cell, PacNIH/A8, based on the neuropathogenic retrovirus A8-V. To construct the expression vector, pA8(Psi-), which expresses the genes gag, pol and env derived from A8-V, the SV40 early region was used for the polyadenylation signal (polyA signal). When a 0.85 kbp fragment in the SV40 early region was employed for the expression vector (pA8(Psi-)beta), env expression was abolished. This abolition was rescued by shortening the SV40 early region to 0.14 kbp (pA8(Psi-)delta). The NHI3T3 cells transfected with pA8(Psi )delta showed expressions of both env and gag genes. PMID- 16377874 TI - Seroprevalence of varicella, measles and hepatitis B among female health care workers of childbearing age. AB - To evaluate the relation between infectious agents and reproductive health hazards for health care workers (HCWs), a cross-sectional study consisting of 73 HCWs and 65 bureau workers was conducted. The reproductive health problems of both groups were compared using a questionnaire, and serologic examinations for measles, varicella and hepatitis B were performed. There were no differences between the two groups according to the rate of seropositivity of measles and varicella (P > 0.05). The prevalence of anti-HBc seropositivity was significantly higher among HCWs than controls (31.5 versus 16.9%). There were no differences between seropositive and seronegative subgroups of measles, varicella and hepatitis B regarding the rates of normal delivery time, preterm and postterm delivery and stillbirth. Subjects seropositive for anti-HBc showed a higher spontaneous abortion rate than those who were seronegative (38.2 versus 16.3, P = 0.009). Although these data showed that HCWs had a high rate of anti-HBc seropositivity and that the rate of spontaneous abortion was associated with past hepatitis B virus infection, further studies including larger populations are needed. We considered that it should be strongly recommended that all HCWs be vaccinated against this virus, and future studies should be focused on the relationship between infectious diseases and reproductive health problems in HCWs. PMID- 16377875 TI - Tuberculous meningitis with multiple intracranial tuberculomas mimicking neurocysticercosis clinical and radiological findings. AB - Central nervous system (CNS) tuberculosis (TB), the most dangerous form of TB, remains a public health problem, particularly in developing countries. In the differential diagnosis of intracranial tuberculomas (ICTs), images on radiological findings should be differentiated from other causes of space occupying lesions. These lesions include malignant diseases such as glioma or lymphoma, pyogenic abscess, toxoplasmosis, neurocysticercosis (NC), sarcoidosis, hydatidosis and late syphilitic involvement of CNS. We present a case with multiple ICTs mimicking NC with similar clinical and imaging manifestations in a young immunocompetent patient. The diagnosis was based on brain magnetic resonance imaging findings. The definitive diagnosis was confirmed mycobacteriologically in cerebrospinal fluid and sputum specimens. Adequate response to anti-TB chemotherapy was achieved while multiple ICTs in the brain disappeared slowly. In the absence of appropriate therapy, these pathologies might be fatal; the possibilities of differential diagnosis would be of great clinical importance, particularly because of the different treatment protocols required for the NC and ICTs. PMID- 16377876 TI - Prevalence of coxsackievirus A5, A6, and A10 in patients with herpangina in Aichi Prefecture, 2005. PMID- 16377877 TI - Norovirus gastroenteritis in Kanagawa Prefecture in December 2004. PMID- 16377878 TI - The molecular epidemiology of ethambutol-resistant Mycobacterium tuberculosis in Henan Province, China. PMID- 16377879 TI - Isolation of influenza virus type AH3 from a traveler returning from Vietnam in July 2005 in Osaka, Japan. PMID- 16377880 TI - Mass food poisoning caused by beef offal contaminated by Escherichia coli O157. PMID- 16377881 TI - An outbreak of Shigatoxin-producing Eshcherichia coli O157:H7 in a nursery school in Mie Prefecture. PMID- 16377882 TI - Secondary transmission of cryptosporidiosis associated with swimming pool use. PMID- 16377883 TI - Two-component phosphorelay signal transduction systems in plants: from hormone responses to circadian rhythms. AB - One of the most fundamental and widespread mechanisms of signal perception/transduction in prokaryotes is generally referred to as the "two component regulatory system (TCS)." The concept of TCS has already been introduced a decade ago from extensive studies on the model prokaryotic bacterium Escherichia coli. Results of recent studies on the model higher plant Arabidopsis thaliana have led us to learn a new scenario as to the versatility of TCS in eukaryotic species. In the plant, on the one hand, TCS are crucially involved in the signal transduction mechanism underlying the regulation of sophisticated plant development in response to hormones (e.g., cytokinin and ethylene). On the other hand, a unique TCS variant is essentially integrated into the plant clock function that generates circadian rhythms, and also tells us the time and season on this regularly spinning and revolving world. In this review, recent progress with regard to studies on TCS in higher plants will be discussed, focusing particularly on the model higher plant Arabidopsis thaliana. PMID- 16377884 TI - Purification and characterization of pyridoxine 5'-phosphate phosphatase from Sinorhizobium meliloti. AB - Here we report the purification and biochemical characterization of a pyridoxine 5'-phosphate phosphatase involved in the biosynthesis of pyridoxine in Sinorhizobium meliloti. The phosphatase was localized in the cytoplasm and purified to electrophoretic homogeneity by a combination of EDTA/lysozyme treatment and five chromatography steps. Gel-filtration chromatography with Sephacryl S-200 and SDS/PAGE demonstrated that the protein was a monomer with a molecular size of approximately 29 kDa. The protein required divalent metal ions for pyridoxine 5'-phosphate phosphatase activity, and specifically catalyzed the removal of Pi from pyridoxine and pyridoxal 5'-phosphates at physiological pH (about 7.5). It was inactive on pyridoxamine 5'-phosphate and other physiologically important phosphorylated compounds. The enzyme had the same Michaelis constant (K(m)) of 385 muM for pyridoxine and pyridoxal 5'-phosphates, but its specific constant [maximum velocity (V(max))/K(m)] was nearly 2.5 times higher for the former than for the latter. PMID- 16377885 TI - Effects of iodine on global gene expression in Saccharomyces cerevisiae. AB - It is well documented that iodine kills microorganisms with a broad spectrum, but a systematic study of its mechanism of action has not yet been reported. Here we found the action of iodine on gene expression level, using the yeast Saccharomyces cerevisiae with a DNA microarray. It was found that, like antimicrobial activity, iodine causes an immediate and dose-dependent (0.5 mM, 0.75 mM and 1 mM) transcriptional alteration in yeast cells. The effects of iodine continued after the first immediate response. Genes for c-compound and carbohydrate metabolism, for energy, and for cell rescue were continuously up regulated. On the other hand, genes related to protein fate were induced especially at 0.5 h. The gene expression profile at 0.5 h was significantly different from that of a longer iodine exposed condition. The main reaction at 0.5 h after iodine addition might be due to oxidative toxicity, and the profile at 0.5 h was similar to that of an agricultural bactericide. PMID- 16377886 TI - Prevention of intestinal infection by glycomacropeptide. AB - The preventive effects of glycomacropeptide (GMP) against intestinal infection were investigated, and conjugates of GMP with xylooligosaccharide (XOS) and carboxymethyldextran (CMD) were prepared by the Maillard reaction to enhance the effect of GMP. The binding ability of GMP to intestinal pathogenic bacteria was evaluated by a binding assay with biotinylated bacteria. GMP showed the ability to bind to Salmonella enteritidis and enterohemorrhagic Escherichia coli O157:H7 (EHEC O157). This binding ability was decreased by a sialidase treatment and completely eliminated by periodate oxidation. These results indicate that such carbohydrate moieties as sialic acid in GMP are involved in binding to S. enteritidis and EHEC O157. The preventive effect of GMP on the adhesion of pathogenic bacteria to Caco-2 cells was also investigated. GMP showed an inhibitory effect on the adhesion of EHEC O157 in a dose-dependent manner, although it was not a potent inhibitor of the adhesion of Salmonella infection. However, in the case of Salmonella infection, GMP-XOS and GMP-CMD significantly suppressed IL-8 production which was the index of infection. Our results indicate GMP to be a promising agent for preventing intestinal infection. PMID- 16377887 TI - Neutralization of Burkholderia pseudomallei protease by Fabs generated through phage display. AB - The isolation of therapeutic and functional protease inhibitors in vitro via combinatorial chemistry and phage display technology has been described previously. Here we report the construction of a combinatorial mouse-human chimeric antibody fragment (Fab) antibody library targeted against the protease of the tropical pathogen, Burkholderia pseudomallei. The resulting library was biopanned against the protease, and selected clones were analyzed for their ability to function as protease inhibitors. Three families of Fabs were identified by restriction fingerprinting, all of which demonstrated high specificity towards the protease of B. pseudomallei. Purified Fabs also demonstrated the capacity to inhibit B. pseudomallei protease activity in vitro, and this inhibitory property was exclusive to the pathogenic protease. Thus these recombinant antibodies are candidates for immunotherapy and tools to aid in further elucidation of the mechanism of action of the B. pseudomallei protease. PMID- 16377888 TI - Spleen cells derived from male non-obese diabetic mice are capable of suppressing the autoantigen-specific production of interferon-gamma of female cells in vitro. AB - Non-obese diabetic (NOD) mice spontaneously develop type 1 diabetes with a strong female predilection. Using co-culture systems with both female and male spleen cells, we found that spleen cells derived from male NOD mice suppress autoantigen (glutamic acid decarboxylase) induced production of interferon-gamma of female cells in vitro. In addition, this suppression appeared to be mediated by a soluble factor(s) produced by male cells in response to the same antigen. Our experimental systems might be useful for further understanding sex differences in autoimmunity as well as in the basic immune response. PMID- 16377889 TI - Cloning and expression of a seed-specific metallothionein-like protein from sesame. AB - A cDNA clone, SiMT encoding an Ec type of metallothionein (MT)-like protein, was isolated from maturing seeds of sesame (Sesamum indicum L.), and its deduced protein sequence shared 47-65% similarity to other known Ec type of MT-like proteins with three highly conserved cysteine-rich segments. The transcript of SiMT was exclusively accumulated in maturing seeds from two weeks after flowering to the end of seed maturation. The results of a southern blot analysis suggested that one SiMT and one SiMT-like gene were present in the sesame genome. Recombinant SiMT fused with glutathione-S-transferase (GST) was over-expressed in Escherichia coli, and purified to homogeneity by affinity chromatography. Recombinant SiMT released from GST was harvested after factor Xa cleavage. Migration of the recombinant SiMT during SDS-PAGE was accelerated when its binding metal ions were depleted by EDTA. The metal-binding capability of recombinant SiMT was measured by inductively-coupled plasma atomic emission spectrometry. Our results show that the recombinant SiMT could trap zinc or copper ions, but not manganese ions, with a stoichiometric ratio (metal ion/SiMT) of approximately 2. PMID- 16377890 TI - Vialinin A, a novel 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenger from an edible mushroom in China. AB - While screening for bioactive compounds from edible mushrooms, a new potent antioxidant, vialinin A (1), together with a known compound, ganbajunin B (2), and a mixture of ganbajunins D (3) and E (4), were isolated from the dry fruiting bodies of Thelephora vialis. The structure of 1, 5',6'-bis(phenylacetoxy) 1,1':4',1''-terphenyl-2',3',4,4''-tetraol, was elucidated by spectroscopic and chemical methods. This compound had strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity with an EC(50) value of 14.0 microM, nearly equal to that of butylated hydroxytoluene (BHT; EC(50) = 10.0 microM). A radical scavenging experiment using 1 and DPPH radicals indicated that 1 donated two hydrogen atoms to two molecules of the DPPH radical under hydrophobic conditions. PMID- 16377891 TI - Identification and overexpression of genes encoding cAMP-dependent protein kinase catalytic subunits in homobasidiomycete Schizophyllum commune. AB - This report describes the first cloning and overexpression experiments on genes encoding cAMP-dependent protein kinase catalytic subunits in homobasidiomycete Schizophyllum commune. We used a degenerate PCR approach to identify two novel genes (ScPKAC1 and ScPKAC2) that are very similar to the catalytic subunits in many eukaryotes. The morphological phenotypes of ScPKAC1 and ScPKAC2 overexpressing clones were compared with those of constitutively active ScGP-A overexpressing clones to determine whether ScPKAC1 and ScPKAC2 are located downstream of heterotrimeric G-protein alpha subunit ScGP-A. Overexpression of constitutively active ScGP-A increased intracellular cAMP levels and suppressed aerial mycelium formation. In contrast, overexpressing ScPKAC1 and ScPKAC2 did not affect the intracellular cAMP levels, though aerial mycelium formation was strongly suppressed. These observations suggest that ScPKAC1 and ScPKAC2 proteins are located downstream of the G-protein alpha subunit ScGP-A in the cAMP signaling pathway. PMID- 16377892 TI - Role of L-histidine in conferring tolerance to Ni2+ in Sacchromyces cerevisiae cells. AB - Ni(2+) toxicity can be alleviated in yeast cells by exogenous L-histidine, but not by its enantiomer, D-histidine, nor by other natural L-amino acids tested. We studied the effect of L-histidine upon the accumulation and intracellular distribution of Ni(2+) and found that moderate L-histidine concentrations (less than or equal to those of Ni(2+)) increased cell tolerance without decreasing Ni(2+) accumulation. Although excess L-histidine appeared to lower Ni(2+) accumulation, the concomitant presence of Ni(2+) and L-histidine in the growth medium stimulated each other's uptake. PMID- 16377893 TI - Screening for new hydroxynitrilases from plants. AB - We established a simple HPLC method to determine the activity and stereochemistry of the chiral mandelonitrile synthesized from benzaldehyde and cyanide, and applied it to screen for hydroxynitrile lyase (HNL) activity of plant origin. A total of 163 species of plants among 74 families were examined for (R)- and (S) HNL activities using the method. We discovered that homogenate of leaves of Baliospermum montanum shows (S)-HNL activity, while leaves and seeds from Passiflora edulis, and seeds from Eriobotrya japonica, Chaenomles sinensis, Sorbus aucuparia, Prunus mume, and Prunus persica show (R)-HNL activity. Partially purified (R)-HNLs from Passiflora edulis and Eriobotrya japonica acted not only on benzaldehyde but also on aliphatic ketone. The enantiomeric excess of (R)-methylpropylketone cyanohydrin synthesized from 2-pentanone using homogenate from leaves of Passiflora edulis was 87.0%, and that of (R)-mandelonitrile synthesized by homogenate from seeds of Eriobotrya japonica was 85.0%. PMID- 16377894 TI - An isolated Candida albicans TL3 capable of degrading phenol at large concentration. AB - An isolated yeast strain was grown aerobically on phenol as a sole carbon source up to 24 mM; the rate of degradation of phenol at 30 degrees C was greater than other microorganisms at the comparable phenol concentrations. This microorganism was further identified and is designated Candida albicans TL3. The catabolic activity of C. albicans TL3 for degradation of phenol was evaluated with the K(s) and V(max) values of 1.7 +/- 0.1 mM and 0.66 +/- 0.02 micromol/min/mg of protein, respectively. With application of enzymatic, chromatographic and mass spectrometric analyses, we confirmed that catechol and cis,cis-muconic acid were produced during the biodegradation of phenol performed by C. albicans TL3, indicating the occurrence of an ortho-fission pathway. The maximum activity of phenol hydroxylase and catechol-1,2-dioxygenase were induced when this strain grew in phenol culture media at 22 mM and 10 mM, respectively. In addition to phenol, C. albicans TL3 was effective in degrading formaldehyde, which is another major pollutant in waste water from a factory producing phenolic resin. The promising result from the bio-treatment of such factory effluent makes Candida albicans TL3 be a potentially useful strain for industrial application. PMID- 16377895 TI - Inhibitory effect of an ellagic acid-rich pomegranate extract on tyrosinase activity and ultraviolet-induced pigmentation. AB - A pomegranate extract (PE) from the rind containing 90% ellagic acid was tested for its skin-whitening effect. PE showed inhibitory activity against mushroom tyrosinase in vitro, and the inhibition by the extract was comparable to that of arbutin, which is a known whitening agent. PE, when administered orally, also inhibited UV-induced skin pigmentation on the back of brownish guinea pigs. The intensity of the skin-whitening effect was similar between guinea pigs fed with PE and those fed with L-ascorbic acid. PE reduced the number of DOPA-positive melanocytes in the epidermis of UV-irradiated guinea pigs, but L-ascorbic acid did not. These results suggest that the skin-whitening effect of PE was probably due to inhibition of the proliferation of melanocytes and melanin synthesis by tyrosinase in melanocytes. PE, when taken orally, may be used as an effective whitening agent for the skin. PMID- 16377896 TI - Characterization, occurrence, and molecular cloning of a lectin from Grifola frondosa: jacalin-related lectin of fungal origin. AB - A lectin named GFL was isolated from the fruiting body of the basidiomycete mushroom Grifola frondosa, which belongs to Aphyllophorales. The lectin had a molecular mass of 24 kDa on SDS-PAGE. The hemagglutinating activity of GFL was not inhibited by any monosaccharide, and inhibited only by porcine stomach mucin so far as tested. The occurrence of GFL was studied at three stages during fruiting body formation. The largest quantity of hemagglutinating activity was found in the fruiting body, and lesser amounts in the mycelial mat and the primordium. The 24-kDa band of GFL was found at all three stages, and the band intensity corresponded to the level of activity in each sample. By cloning and sequencing the GFL-cDNA, the primary structure of this lectin was determined. GFL is composed of 181 amino acids, having no signal peptide. The amino acid sequence was found to be homologous to those of so-called jacalin-related plant lectins, suggesting that GFL is the first example of a jacalin-related lectin of fungal origin. PMID- 16377897 TI - Effects of culture conditions on ergosterol biosynthesis by Saccharomyces cerevisiae. AB - Ergosterol is an essential component of yeast cells that maintains the integrity of the membrane. It was investigated as an important factor in the ethanol tolerance of yeast cells. We investigated the effects of brewing conditions on the ergosterol contents of S. cerevisiae K-9, sake yeast, several kinds of Saccharomyces cerevisiae that produce more than 20% ethanol, and X2180-1A, laboratory yeast. K-9 had a higher total ergosterol contents under all the conditions we examined than X2180-1A. Ethanol and hypoxia were found to have negative and synergistic effects on the total ergosterol contents of both strains, and significantly reduced the free ergosterol contents of X2180-1A but only slightly reduced those of K-9. The maintenance of free ergosterol contents under brewing conditions might be an important character of sake yeast strains. DNA microarray analysis also showed higher expression of ergosterol biosynthesis genes in K-9 than in X2180-1A. PMID- 16377898 TI - Functions of family-22 carbohydrate-binding modules in Clostridium josui Xyn10A. AB - Clostridium josui xylanase Xyn10A is a modular enzyme comprising two family-22 carbohydrate-binding modules (CBMs), a family-10 catalytic module (CM), a family 9 CBM, and two S-layer homologous modules, consecutively from the N-terminus. To study the functions of the family-22 CBMs, truncated derivatives of Xyn10A were constructed: a recombinant CM polypeptide (rCM), a family-22 CBM polypeptide (rCBM), and a polypeptide composed of the family-22 CBMs and CM (rCBM-CM). Recombinant proteins were characterized by enzyme and binding assays. rCBM-CM showed the highest activity toward xylan and weak activity toward some polysaccharides such as barley beta-glucan and carboxymethyl-cellulose. Although rCBM showed an affinity for insoluble and soluble xylan as well as barley beta glucan and Avicel in qualitative binding assays, removal of the CBMs negligibly affected the catalytic activity and thermostability of the CM. PMID- 16377899 TI - Probing the secondary structure of a recombinant neuronal adaptor protein and its phosphotyrosine binding domains. AB - Rat brain Fe65 and its truncated forms corresponding to the combined PTB1 and PTB2 domains, as well as to the isolated PTB2 domain, were expressed in Escherichia coli and purified from inclusion bodies by affinity chromatography. The recombinant proteins were refolded and judged functionally active by their ability to interact with native APP. Limited proteolysis of recombinant Fe65 and PTB1-2 with trypsin, chymotrypsin and V8 proteases showed that the most sensitive proteoltytic sites were positioned at the level of the interdomain regions comprised between WW/PTB1 and PTB1/PTB2. Secondary structure of the recombinant proteins, evaluated by CD spectroscopy, showed a different degree of unordered structures, the PTB2 domain being the higher organised region. In addition, intrinsic fluorescence measurements of PTB2, indicated that a conformational transition of the protein can be induced by denaturating agents such as GuHCl. These data provide first evidences on the secondary structural levels of Fe65. PMID- 16377900 TI - Synthesis of the four stereoisomers of 2,6-dimethyloctane-1,8-dioic acid, a component of the copulation release pheromone of the cowpea weevil, Callosobruchus maculatus. AB - A diastereomeric mixture and the four stereoisomers of 2,6-dimethyloctane-1,8 dioic acid (2), a copulation release pheromone of the cowpea weevil, Callosobruchus maculatus, were synthesized. The stereoisomeric purities of the four synthetic isomers of 2 were determined by the HPLC analyses of their bis-2 (2,3-anthracenedicarboximide)-1-cyclohexyl esters. PMID- 16377901 TI - Combined effects of dietary protein type and fat level on the body fat-reducing activity of conjugated linoleic acid (CLA) in rats. AB - The interaction of dietary protein type and fat level on the body fat-reducing activity of conjugated linoleic acid (CLA) was studied in male rats fed diets containing casein (CAS) or soy protein (SOY) as a protein source with low fat (LF, 6.0% soybean oil) or high fat (HF, 13.0% soybean oil) combinations for 4 weeks. CLA was added at the 1.0% level to all diets. The weight of perirenal adipose tissue tended to be lower in the SOY groups than in the corresponding CAS groups, and the difference between the LF diets was significant. The weight of epididymal adipose tissue showed a similar but insignificant trend. The weight of brown adipose tissue was heaviest on the SOY-HF diet and lowest on two CAS diets, the SOY-LF diet being intermediate. The concentration of serum leptin was lowest on the SOY-LF diet and was significantly lower than that of the corresponding CAS group, but this difference disappeared when the dietary fat level increased. The serum cholesterol-lowering activity of SOY in relation to CAS was reproduced even when CLA was given. Thus the body fat-reducing activity of CLA was most marked when rats were fed the SOY-LF diet. Although the CAS-HF diet increased body fat deposition, the magnitude of the reduction by lowering dietary fat level was more marked than in the case of SOY. These results indicate a complicated interaction of dietary manipulations with the body fat-reducing effect of CLA, but the combination of CLA with the SOY-LF diet appears to be an appropriate approach. PMID- 16377902 TI - Identification of novel decenoic acids in heated butter. AB - Novel decenoic acids such as (E)-4-decenoic acid and (E)- and (Z)-5-,6-decenoic acid were detected as minor components in heated butter using GC and GC/MS. The formation mechanism of these novel decenoic acids is discussed on the basis of the result of the reaction of delta-decalactone with active clay in a model experiment. PMID- 16377903 TI - Isolation and functional analysis of cytochrome P450 CYP153A genes from various environments. AB - The cytochrome P450 CYP153 family is thought to mediate the terminal hydroxylation reactions of n-alkanes. We isolated 16 new P450 CYP153A genes (central region) from various environments such as petroleum-contaminated soil and groundwater, as well as one from the n-alkane-degrading bacterium Alcanivorax borkumensis SK2 (designated P450balk). The sequences of the new P450 genes were extended by PCR to generate full-length chimeric P450 genes, using the N- and C terminal domains of P450balk. A differential CO-reduced P450 spectral analysis indicated that 8 P450 genes among the 16 chimeric genes were expressed in Escherichia coli to generate a soluble and functional enzyme. The several functional chimeric P450s and P450balk were further fused to the reductase domain of the self-sufficient P450 monooxygenase (P450RhF) at the C-terminus. E. coli cells expressing these self-sufficient P450 chimeric genes converted n-alkanes, cyclohexane, 1-octene, n-butylbenzene, and 4-phenyl-1-butene into 1-alkanols, cyclohexanol, 1,2-epoxyoctane, 1-phenyl-4-butanol, and 2-phenethyl-oxirane, respectively. PMID- 16377904 TI - Expression of a chemorepellent factor, Slit2, in peripheral nerve regeneration. AB - The expression of mRNA for chemorepellent factors slit1 and slit2 in rat peripheral nerve regeneration was examined. The mRNA of slit2 increased when the continuity of basal lamina tubes was disrupted, not when it remained and the Slit2 protein was located in Schwann cells. These results suggest that disruption of the continuity of basal lamina tubes induces the expression of slit2 in Schwann cells during peripheral nerve regeneration. PMID- 16377905 TI - Cloning and sequencing of the aldehyde oxidase gene from Methylobacillus sp. KY4400. AB - The aldehyde oxidase genes (aods) from Methylobacillus sp. KY4400 were cloned, and sequenced. The sequences for small (aodS, 489 bp), medium (aodM, 993 bp), and large (aodL, 2,328 bp) subunit genes were determined. At least one additional ORF was indispensable for the expression of enzyme activity. The structural genes contained two [2Fe-2S] centers, an FAD binding site, and a molybdenum cofactor binding site. PMID- 16377906 TI - Dietary isothiocyanates modify mitochondrial functions through their electrophilic reaction. AB - We found that both benzyl isothiocyanate (ITC) and phenyl ITC inhibited respiration in the mitochondria in an electrophilic reaction-dependent manner. ITC-induced mitochondrial swelling and cytochrome c release were prevented by cyclosporin A, indicating that they are mediated through the ITC moiety-dependent reaction to critical thiol groups for the opening of membrane permeability transition-dependent pores. PMID- 16377907 TI - Effect of wheat gluten hydrolysate on the immune system in healthy human subjects. AB - Nine healthy volunteers were divided into a test group (n = 5) and a control group (n = 4). The test group consumed 3 grams per d of wheat gluten hydrolysate for 6 d, and their NK cell activity and hematological parameters were measured: The same assessments were performed in the control group, which did not receive wheat gluten hydrolysate. In the test group, NK cell activity increased significantly (P = 0.018) after wheat gluten hydrolysate intake. No adverse effects were observed in either group. PMID- 16377908 TI - GLR1 plays an essential role in the homeodynamics of glutathione and the regulation of H2S production during respiratory oscillation of Saccharomyces cerevisiae. AB - The role of glutathione (GSH) and its homeodynamics during respiratory oscillation of Saccharomyces cerevisiae were investigated. Pulse injection of thiol redox modifying agents, such as diethylmaleate, N-ethylmaleimide, DL butione-[S,R]-sulfoxamine, or 5-nitro-2-furaldehyde into the culture perturbed oscillation, although the degree of perturbation varied. Analysis of the expression profiles of GSH1 and GLR1, the activities of glutathione reductase, oscillations in cysteine and GSH concentrations, and the chemostat culture of the GLR1 disruptant indicated that GLR1 plays an essential role in the homeodynamics of GSH and the regulation of H(2)S production. PMID- 16377909 TI - Heat-epimerized tea catechins have the same cholesterol-lowering activity as green tea catechins in cholesterol-fed rats. AB - Tea catechins are known to be epimerized by heat treatment. The effect of heat epimerized tea catechins on serum cholesterol concentration was compared with that of green tea catechins. Our observations strongly suggest that both tea catechins and heat-epimerized tea catechins lower serum cholesterol concentration by inhibiting cholesterol absorption in the intestine. There was no differential effect between the two catechin preparations. PMID- 16377910 TI - Production of completely flavinylated histamine dehydrogenase, unique covalently bound flavin, and iron-sulfur cluster-containing enzyme of nocardioides simplex in Escherichia coli, and its properties. AB - The hmd gene of histamine dehydrogenase from Nocardioides simplex was overexpressed in Escherichia coli, and the resulting enzyme was purified to homogeneity. The purified recombinant enzyme is almost identical with the native enzyme in view of molecular weight and specific activity, and is stoichiometrically assembled with the three cofactors 6-S-cysteinyl FMN, 4Fe-4S cluster, and ADP. PMID- 16377911 TI - Development of a modified positive selection medium that allows to isolate Aspergillus oryzae strains cured of the integrated niaD-based plasmid. AB - The nitrate reductase gene (niaD) is the most frequently utilized as a selectable marker for homologous integration at the niaD locus of Aspergillus oryzae. In this study we developed a method for curing of the niaD-based plasmid integrated on the A. oryzae genome. Positive selection using a modified chlorate medium containing leucine as a nitrogen source enabled efficient isolation of the strains deficient in nitrate assimilation from the niaD(+) transformant. PCR analysis of the strains confirmed that the homologously integrated plasmid carrying the h2b-egfp fusion gene was cured by intrachromosomal recombination which was accompanied by the loss of the EGFP-fluorescence. PMID- 16377912 TI - In vitro augmentation of natural killer activity and interferon-gamma production in murine spleen cells with Agaricus blazei fruiting body fractions. AB - Aqueous extracts of the Agaricus blazei fruiting body prepared at different temperatures were fractionated by ethanol precipitation with various ethanol concentrations. The original aqueous extracts of A. blazei failed to stimulate natural killer (NK) cell activity in murine spleen cells in vitro, but the strongest effect was observed in a 30% ethanol-soluble-50% ethanol-insoluble fraction prepared from the extract at 40 degrees C (fraction A-50). Fraction A-50 also showed the strongest augmenting effect on interferon (IFN)-gamma production. This augmentation of NK activity and IFN-gamma production by fraction A-50 was completely abrogated by a heat treatment. PMID- 16377913 TI - Inhibition of preadipocyte differentiation by germacranolides from Calea urticifolia in 3T3-L1 cells. AB - The effects of germacranolides isolated from Calea urticifolia on adipocytic differentiation of 3T3-L1 cells were examined. These germacranolides inhibited adipogenesis at a concentration of 1.25-5 microM. But no inhibitory activity against cell proliferation and no nonspecific binding activity to protein were observed. These results indicate that these germacranolides are the specific inhibitors of preadipocyte differentiation. PMID- 16377914 TI - Isolation and characterization of glucose derepressed invertase mutants from Schizosaccharomyces pombe. AB - We have isolated 14 different Schizosaccharomyces pombe mutants that synthesize invertase enzyme constitutively. Analyses of invertase activities revealed that the degrees of resistance to glucose repression were not similar among different complementation groups. One of the complementation groups appeared to be associated with functional and/or regulatory defects in hexose transport. Another complementation group appeared to be specific for the regulation of the inv1 gene alone, implying that these mutations might be associated with different genes acting on the glucose sensing and signaling pathway. In addition, we found that the wild-type level glucose uptake is essential for the full-level repression of inv1 expression. PMID- 16377915 TI - Lifespan extending activity of substances secreted by the nematode Caenorhabditis elegans that include the dauer-inducing pheromone. AB - The nematode Caenorhabditis elegans yields a substance(s) inducing the larval diapause, called dauer-inducing pheromone. We discovered that the crude pheromone extract extends the adult lifespan in the animal. This extension does not occur in the mutant animal, in which expansion of the lifespan caused by other mutations reducing insulin signaling is suppressed. This is the first description concerning the relevancy of the pheromone to the longevity in the animal. PMID- 16377916 TI - Effects of nocturnal oxygen therapy on outcome measures in patients with chronic heart failure and cheyne-stokes respiration. AB - BACKGROUND: The effects of nasal oxygen (O(2)) supply at night using conventional home oxygen therapy (HOT) equipment on quality of life (QOL) and sleep-disordered breathing (SDB) were evaluated in patients with congestive heart failure (CHF). Nasal nocturnal O(2) therapy not only stabilizes SDB but also reduces sympathetic activity, and improves exercise capacity in patients with CHF. However, the effects of oxygen on the cardiac function and QOL of heart failure patients have not been fully elucidated. METHODS AND RESULTS: Fifty-six patients with CHF (New York Heart Association class II - III, left ventricular ejection fraction (LVEF) or=2.5 mm. METHODS AND RESULTS: A retrospective survey of the outcome of coil occlusions for PDA >or=2.5 mm before and after the 0.052-inch coil became available found that (1) the frequency of PDA >or=2.5 mm among all candidates for coil occlusion significantly increased after the availability of a 0.052-inch coil (p<0.01); (2) deployment complicated by migration (p<0.01), and prolonged procedure time (p<0.05) were significantly decreased after the introduction of the 0.052-inch coil. In a multivariate logistic regression model for uneventful deployment adjusted for age, pulmonary to systemic flow ratio, and use of a 0.052-inch coil, use of the 0.052-inch coil significantly decreased eventful deployment (p<0.05); and (3) successful deployment of a coil for PDA >or=4 mm significantly increased with the 0.052-inch coil (p<0.01). Complete occlusion was achieved once deployment was successful. CONCLUSION: Introduction of the 0.052-inch coil decreased complicated coil occlusion deployment for PDA >or=2.5 mm, and contributed to a better likelihood of coil occlusion for PDA >or=4 mm. PMID- 16377921 TI - Platelet activity is a biomarker of cardiac necrosis and predictive of untoward clinical outcomes in patients with acute myocardial infarction undergoing primary coronary stenting. AB - BACKGROUND: The relationship between platelet activity and myocardial injury in patients with ST-segment elevated (ST-se) acute myocardial infarction (AMI) remains unclear. This study tested the hypothesis that platelet activity (expressed by CD62p) is enhanced and predictive of both the extent of myocardial damage and 30-day clinical outcome in patients with ST-se AMI undergoing primary coronary stenting. METHODS AND RESULTS: Platelet CD62p expression prior to coronary angiographic was prospectively measured using flow cytometry in 45 consecutive patients with AMI undergoing primary coronary stenting. The CD62p expression was also evaluated in 20 healthy and 20 at-risk control subjects. The CD62p expression was significantly higher in AMI patients than in healthy and at risk control subjects (all p values <0.0001). Patients with high CD62p expression (>or=8%) had significantly higher creatine kinase-MB (p<0.0001) levels, higher incidence of cardiogenic shock (p=0.009) upon presentation, significantly lower left ventricular ejection fraction (p=0.0003), and significantly higher incidence of 30-day composite major adverse clinical outcomes (MACO) (advanced congestive heart failure >or=class 3 or 30-day mortality) (p<0.0001) than those patients with low CD62p expression (<8%). Multiple stepwise logistic regression analysis demonstrated that only high CD62p expression (>or=8%) was an independent predictor of 30-day MACO (all p<0.0001). CONCLUSIONS: Platelet activation was significantly increased in patients with ST-se AMI. Initial CD62p expression was independently associated with extent of myocardial damage and 30-day MACO. PMID- 16377922 TI - Effects of intraaortic balloon pumping on the angiographic no-reflow phenomenon after percutaneous coronary intervention in patients with anterior myocardial infarction. AB - BACKGROUND: The effects of intraaortic balloon pumping (IABP) are considered to be a reduction in myocardial oxygen demand because of systolic left ventricular unloading and an increase in coronary blood flow. Although the former effect has been consistently recognized, the latter effect remains controversial. The purpose of this study was to examine the effects of IABP on the angiographic no reflow phenomenon. METHODS AND RESULTS: The coronary flow velocity pattern of the left anterior descending artery (LAD) was assessed by transthoracic Doppler echocardiography during IABP counterpulsation, and the effects of IABP were compared between angiographic no-reflow and good reflow patients. The study group comprised 17 patients with anterior myocardial infarction who underwent percutaneous coronary intervention and IABP for typical clinical indications. Echocardiographic data were obtained during 2:1 balloon pumping after coronary angioplasty. In the good reflow group (n=9), IABP counterpulsation increased the mean diastolic flow velocity (MDFV) and peak diastolic flow velocity (PDFV) by 56 +/- 32% (p<0.001) and 48 +/- 27% (p<0.001), respectively. In contrast, in the no reflow group (n=8), IABP only increased the MDFV and PDFV by 19 +/- 33% (p=0.24) and 6 +/- 12% (p=0.22), respectively. Diastolic deceleration time was smaller and the prevalence of systolic retrograde flow was greater in the no-reflow group than in the good reflow group, and IABP affected neither parameter. CONCLUSIONS: IABP had limited effects on LAD flow velocity pattern in patients with the angiographic no-reflow phenomenon. PMID- 16377923 TI - Radiation exposure to patient's skin during percutaneous coronary intervention for various lesions, including chronic total occlusion. AB - BACKGROUND: Radiation skin injuries have been reported as a result of various procedures, so in the present study the patients' entrance skin dose (ESD) during percutaneous coronary intervention (PCI) was evaluated. METHODS AND RESULTS: ESDs were assessed during 97 procedures (13 for chronic total occlusion (CTO), 14 for multivessel stenoses, 22 for single-vessel multiple stenoses, and 48 for single stenosis). The patients wore jackets that had 48 or 52 radiosensitive indicators placed on the back during the PCI procedures, with 8 other indicators placed on both upper arms. After the procedure, the color of the indicators was analyzed with a color measuring instrument, and the patients' ESDs were calculated from the color difference of the indicators. The average maximum ESDs of the patients were 4.5 +/- 2.8 Gy (median: 4.6 Gy) for CTO, 2.3 +/- 0.7 Gy (median: 2.4 Gy) for multivessel stenoses, 1.8 +/- 1.0 Gy (median: 1.5 Gy) for single-vessel multiple stenoses, and 1.4 +/- 0.9 Gy (median: 1.2 Gy) for single stenosis. CONCLUSIONS: Skin injury can occur during PCI, especially for CTO, so it is important to estimate each patient's ESD and attempt to reduce it. PMID- 16377924 TI - Comparable prognostic value of vasodilator response to acetylcholine in brachial and coronary arteries for predicting long-term cardiovascular events in suspected coronary artery disease. AB - BACKGROUND: Vasodilator response to acetylcholine (ACh) (ie, endothelium dependent dilation) is impaired in the peripheral and coronary circulation of patients with coronary risk factors and coronary artery disease (CAD). There is a close relationship of vasodilator response to ACh in both the coronary artery (CA) and the brachial artery (BA), but the comparative prognostic importance of these responses has not been fully investigated in relatively low-risk suspected CAD. METHODS AND RESULTS: The flow responses of both the CA and BA were measured in 70 patients with suspected CAD, excluding patients with triple-vessel disease and known peripheral or cerebrovascular disorders. A Doppler guidewire was placed into a major branch of the CA and a proximal portion of the left BA. ACh was infused at 10(-8), 10(-7) and 10(-6) mol/L for 3 min into the CA and at 7.5, 15, and 30 mug/min for 5 min into BA. The flow response was obtained by multiplying the average peak velocity by the cross-sectional area from quantitative angiography. Vasodilator response to ACh was assessed by the ratio of ACh-induced flow/baseline flow, expressed as coronary blood flow index (CBFI) or brachial blood flow index (BBFI). There were 39 CAD patients (61 +/- 8 years old) and 31 normal coronary patients (NL, 58 +/- 11 years old) who were followed up for 53 +/ 17 months. Eleven patients had coronary events (CE) during this period: 1 case of nonfatal myocardial infarction and 10 cases of unstable angina. A strong correlation between CBFI and BBFI was observed at middle-and high-doses of ACh (r=0.72, p<0.0001, 15 microg/min vs 10(-7) mol/L; r=0.76, p<0.0001, 30 microg/min vs 10(-6) mol/L). Kaplan-Meier analysis, using the best cut-off values obtained from receiver-operating characteristic curves for CE, revealed that both CBFI and CAFI were significant predictors for CE. CONCLUSIONS: The BA vasodilator response to optimal ACh dosage can be used as a surrogate prognostic predictor for coronary endothelial function tests in patients with suspected CAD. PMID- 16377925 TI - Prolonged QRS duration and severity of mitral regurgitation are unfavorable prognostic markers of heart failure in patients with nonischemic dilated cardiomyopathy. AB - BACKGROUND: The goal of the present study was to identify predictors of event free survival in nonischemic dilated cardiomyopathy (NIDCM) patients after administration of angiotensin-converting enzyme inhibitors (ACEI)/angiotensin receptor blockers (ARB) and beta-blockers. METHODS AND RESULTS: The study group comprised 78 consecutive patients with NIDCM between 1997 and 2002. NIDCM was defined as ejection fraction (EF) <0.40 and left ventricular end-diastolic diameter (LVEDD) >55 mm on echocardiography and normal coronary angiography. The mean EF and LVEDD was 26.3 +/- 10.5%, and 62.9 +/- 7.1 mm, respectively. Patients were treated with optimal medical therapy including ACEI/ARBs and/or beta blockers and followed up for 35.6 +/- 27.8 months. The primary endpoint was either cardiac death or hospitalization because of deterioration of heart failure. Cox's regression analysis was used to establish the association of age, sex, EF, LVEDD, left atrial diameter, cardiac index, pulmonary capillary wedge pressure, QRS duration, severity of mitral regurgitation, body mass index, New York Heart Association class and the presence of atrial fibrillation with these events. During follow-up, 23 patients reached the primary endpoint. In a multivariate analysis, EF (chi-square 5.74, p=0.0166), severity of mitral regurgitation (chi-square 12.31, p=0.0004), and QRS duration (chi-square 11.20, p=0.0008) remained significant predictors. CONCLUSION: In NIDCM patients, prolonged QRS duration is a high risk factor for remodeling and unfavorable events. The severity of mitral regurgitation was also a strong risk predictor. PMID- 16377926 TI - Effects of dual-chamber pacing on regional myocardial deformation in patients with hypertrophic obstructive cardiomyopathy. AB - BACKGROUND: This study examined the effects of dual-chamber pacing (DDD) on regional myocardial deformation, as determined by echocardiographic strain and strain rate (SR) imaging, in patients with hypertrophic obstructive cardiomyopathy (HOCM). METHODS AND RESULTS: Fourteen patients (11 men, 3 women; mean age 55 +/-16 years) who had been on long-term DDD (mean period 7.4 +/- 2.1 years) underwent strain and SR imaging. Before and after DDD, the peak strain (%) and SR (s(-1)) during systole were assessed in 8 segments in 4 left ventricular (LV) walls. With DDD turned on, peak strain and SR were significantly increased in the basal anteroseptal (strain -10.2 +/- 6.8 to -1.0 +/- 6.4, p<0.005; SR 0.76 +/- 0.46 to 0.05 +/- 0.58, p<0.001) and septal segments (strain -11.2 +/- 8.9 to -2.2 +/- 7.7, p<0.005; SR -0.85 +/- 0.54 to -0.19 +/- 0.75, p<0.05), but not in the basal posterior (strain -15.0 +/- 13.0 to -13.4 +/- 9.2, p=NS; SR 1.37 +/- 0.57 to -1.93 +/- 0.65, p=NS) and lateral segments (strain -18.1 +/- 10.2 to -15.7 +/- 5.6, p=NS; SR -1.33 +/- 0.68 to -0.84 +/- 0.88, p=NS). These findings were associated with a modest, but significant, change in the LV pressure gradient (24 +/- 12 mmHg to 14 +/- 7 mmHg, p<0.001). CONCLUSIONS: In patients with HOCM, DDD appeared to produce myocardial lengthening in the basal septum during systole, which may have implications for the mechanism of reducing LV outflow obstruction during DDD. PMID- 16377927 TI - Evaluation of arrhythmogenic substrate in patients with hypertrophic cardiomyopathy using wavelet transform analysis. AB - BACKGROUND: Hypertrophic cardiomyopathy (HCM) is thought to have a microvolt level electrical disarrangement in the myocardium that leads to ventricular tachyarrhythmias and sudden cardiac death. Although signal-averaged electrocardiography (ECG) has been used to detect late potential as a parameter of electrical instability, its predictability is not high. The focus of the present study was the ability of high-resolution wavelet transform from beat-to beat analysis to detect arrhythmogenic substrates and to evaluate its relationship to the severity of ventricular tachycardia. METHODS AND RESULTS: The study group comprised 50 healthy subjects and 50 patients with HCM. The filtered QRS duration from the signal-averaged ECG, the high-power duration (HPD) and number of disarrangement points (NDP) from the wavelet-transform ECG were measured. When HPD was defined >114 ms and/or NDP >9 points as abnormal, the sensitivity and specificity for ventricular tachycardia was 93.8% and 79.4%, respectively. When a mean +/- standard deviation of the HPD in normal subjects was defined as normal, 93.8% of patients with a positive late potential were out of the normal range. CONCLUSIONS: The newly developed color-display 3-dimensional wavelet transform system showed good time-frequency resolution in analyzing every single beat without signal-averaging. The analysis could be used to detect arrhythmogenic substrates in patients with HCM. PMID- 16377928 TI - Low-frequency electrical stimulation increases muscle strength and improves blood supply in patients with chronic heart failure. AB - BACKGROUND: This study was designed to evaluate the effects of low-frequency electrical stimulation (LFES) on muscle strength and blood flow in patients with advanced chronic heart failure (CHF). METHODS AND RESULTS: Patients with CHF (n=15; age 56.5 +/- 5.2 years; New York Heart Association III - IV; ejection fraction 18.7 +/- 3.3%) were examined before and after 6 weeks of LFES (10 Hz) of the quadriceps and calf muscles of both legs (1 h/day, 7 days/week). Dynamometry was performed weekly to determine maximal muscle strength (F(max); N) and isokinetic peak torque (PT(max); Nm); blood flow velocity (BFV) was measured at baseline and after 6 weeks of LFES using pulsed-wave Doppler velocimetry of the right femoral artery. Six weeks of LFES significantly increased F(max) (from 224.5 +/- 96.8 N to 340.0 +/- 99.4 N; p<0.001), and also PT(max) (from 94.5 +/- 41.5 Nm to 135.3 +/- 28.8 Nm; p<0.01). BFV in the femoral artery increased after 6 weeks from 35.7 +/- 15.4 cm/s to 48.2 +/- 18.1 cm/s (p<0.05); BFV values at rest before and after 6 weeks of LFES did not differ significantly. CONCLUSIONS: LFES may improve muscle strength and blood supply, and could be recommended for the treatment of patients with severe CHF. PMID- 16377929 TI - Polymorphism of methylenetetrahydrofolate reductase gene (C677T MTHFR) is not a confounding factor of the relationship between serum uric acid level and the prevalence of hypertension in Japanese men. AB - BACKGROUND: The association between serum uric acid (UA) and the prevalence of hypertension, and the relationship between methylenetetrahydrofolate reductase (MTHFR) polymorphism and hypertension remains unclear. The aim of the present study was to investigate whether the C677T MTHFR mutation genotype (VV) is independently associated with the prevalence of hypertension or blood pressure (BP), and examined any interaction of MTHFR and UA with BP. METHODS AND RESULTS: Participants were randomly selected from all residents (aged 40-69 years) in a rural county of Japan, and the data for the men (n=335) were analyzed. ;Hypertension' was defined as systolic BP >or=140 and/or diastolic BP >or=90 mmHg and/or being administered antihypertensive medication. Serum UA level was independently associated with the prevalence of hypertension (odds ratio (95% confidence interval) =2.7 (1.2-5.9), p=0.047) for the highest tertile of serum UA (>or=398.5 micromol/L (6.7 mg/dl)) vs that of the lowest tertile (<321.2 micromol/L (5.4 mg/dl)), but the MTHFR mutation was not independently associated with prevalence of hypertension or BP. No interaction of the MTHFR mutation and serum UA with BP was found. CONCLUSIONS: The mutation of C677T MTHFR was not independently associated with the prevalence of hypertension or BP levels although serum UA was. Furthermore, the relationship between serum UA and BP was not modulated by the MTHFR mutation in Japanese men. PMID- 16377930 TI - New approach to pulmonary vein isolation for atrial fibrillation using a multielectrode basket catheter. AB - BACKGROUND: Pulmonary vein (PV) isolation using a circular catheter creates an entrance block from the left atrium (LA) to the PV, which eliminates paroxysmal atrial fibrillation (PAF). A new approach to PV isolation during distal PV pacing is to use a basket catheter. METHODS AND RESULTS: Fifty consecutive patients with PAF underwent basket-catheter-guided PV isolation. PV pacing was performed from the distal electrode pair of the basket catheter. The exit breakthrough point was targeted for segmental PV isolation. The endpoint was the elimination of bidirectional PV - LA conduction. A repeat ablation procedure was performed in 12 of 14 patients who had recurrence of AF. The recovery of PV -LA conduction was noted in 24 of the 48 PVs, and 5 PVs (21%) had unidirectional block. At 12 months, 80% of patients were free of AF without antiarrhythmic drugs. No PV stenosis >50% was detected at 12 months after the procedure. CONCLUSIONS: This new approach for PV isolation during distal PV pacing using a basket catheter is useful for confirming bidirectional PV - LA conduction block. PV isolation that creates not only an entrance block but also an exit block at the PV - LA junction may be required to cure paroxysmal AF. PMID- 16377931 TI - Prediction of paroxysmal atrial fibrillation using nonlinear analysis of the R-R interval dynamics before the spontaneous onset of atrial fibrillation. AB - BACKGROUND: New methods based on nonlinear theory have been developed to give more insight into complex heart rate (HR) dynamics. This study was designed to test the hypothesis that altered HR dynamics, as analyzed with complexity and fractal measures, may precede the spontaneous onset of paroxysmal atrial fibrillation (PAF). Secondly, the difference in the temporal change of these measurements between the different types of atrial fibrillation (AF) was assessed. METHODS AND RESULTS: From 105 Holter tapes in which PAF was recorded, 44 PAF (>or=5 min) episodes in 33 patients (22 men, 58 +/- 12 years), preceded by sinus rhythm for more than 1 h, were selected and submitted to time-and frequency domain HR variability analyses, along with detrended fluctuation analysis, approximate entropy (ApEn) and sample entropy (SampEn). The 60 min before the onset of AF were divided into 6 10-min periods and studied using repeated measures ANOVA. PAF episodes were divided into 3 subgroups: an increased HF component and decreased L/H ratio (vagal type, n=20); increased L/H ratio and decreased HF component (sympathetic type, n=14); and non-related type (n=10). None of the time- or frequency-domain parameters showed any significant change before AF in any type of AF. The alpha(1) showed a tendency to decrease before the onset of AF and the change in alpha(1) was divergent according to the AF type. The ApEn decreased before the onset of AF (1.005+/-0.046, 60-50 min before AF to 0.894+/-0.052, 10-0 min before AF; p=0.032). The SampEn also decreased progressively before the start of AF (1.165 +/- 0.085, 60-50 min before AF to 0.887 +/- 0.077, 10-0 min before AF, p=0.003). The decrease in both the ApEn and SampEn was irrespective of the AF type. CONCLUSIONS: A reduction in the ApEn and SampEn, which reflects the nonlinear complexity of HR variability, is a hallmark of altered HR dynamics preceding the spontaneous onset of AF. PMID- 16377932 TI - Determinants of brain natriuretic peptide levels in patients with lone atrial fibrillation. AB - BACKGROUND: Although brain natriuretic peptide (BNP) is increasingly being used for screening and monitoring of congestive heart failure, its utility in patients with lone atrial fibrillation (AF) is unclear. METHODS AND RESULTS: Plasma BNP levels were measured and comprehensive transthoracic echocardiography was performed in 96 subjects (47: sinus rhythm, 49: AF). Patients with structural heart disease were excluded. Potential determinants of BNP levels were identified by univariate and multivariate analyses. Individuals with AF had higher BNP levels than those with sinus rhythm (150 +/- 114 vs 49 +/- 61 pg/ml, p<0.001) The left atrial (LA) volume index (r=0.63, p<0.001), the pulmonary artery systolic pressure (r=0.45, p=0.006), and the early mitral inflow velocity (E)/mitral annular velocity (E') (r=0.36, p=0.04) were found to be independently correlated with BNP level. The correlations between BNP level and LA volume index (p=0.001) or E/E' (p=0.03) were unaltered when subjects with sinus rhythm were removed from the analysis. CONCLUSIONS: BNP levels significantly correlated with LA volume index and E/E' in patients with lone AF, which indicates that the BNP level may reflect early left ventricular dysfunction and LA enlargement in this patient population. PMID- 16377933 TI - Image fusion of coronary tree and regional cardiac function image using multislice computed tomography. AB - BACKGROUND: Although trials of image fusion, such as positron emission computed tomography and multislice spiral computed tomography (MSCT), have already demonstrated clinical usefulness, fusion of the coronary artery image and functional image by MSCT alone has not been reported yet. Here, a new idea of data analysis is proposed in which both regional cardiac function and the responsible coronary arteries can be assessed by a fused image. METHODS AND RESULTS: The study group comprised 5 patients with coronary artery disease. At the first procedure, 3 dimensional (D) volume rendering coronary artery (3D-CTA) was extracted. At the second procedure, the systolic regional wall thickening was calculated and the color 3D functional surface map of systolic wall thickening (3D-SWT) was generated. At the final procedure, 3D-SWT was superimposed on the left ventricular surface with 3D-CTA using a transparency. In all 5 patients, image fusion of the coronary tree and cardiac function was correctly generated. Image fusion can be displayed as clear 3D images, offering better orientation to help assess both the coronary artery and regional function. CONCLUSIONS: Image fusion of coronary computed tomography angiography and the functional map by MSCT is potentially a new method of assessing both the coronary artery and cardiac function. PMID- 16377935 TI - Short duration of reactive hyperemia in the forearm of subjects with multiple cardiovascular risk factors. AB - BACKGROUND: Peripheral vascular endothelial dysfunction is an independent predictor of cardiovascular events, and can be assessed noninvasively by measuring reactive hyperemia, either by vascular ultrasound measurement of flow mediated vasodilatation or, less commonly, by measurement of blood flow using plethysmography. In the present study reactive hyperemia was measured using plethysmography in healthy subjects with multiple cardiovascular risk factors. METHODS AND RESULTS: Reactive hyperemia was measured following 5-min occlusion of the upper arm in 449 healthy subjects (302 men, 147 women, age range 20-70 years) with (n=352) and without (n=97) risk factors such as smoking, hypertension, diabetes mellitus, hypercholesterolemia, obesity, family history of cardiovascular disease, and menopause. Maximum blood flow and minimum vascular resistance in reactive hyperemia did not differ between subjects with and without risk factors regardless of gender. Duration of reactive hyperemia, however, was significantly shorter in subjects with risk factors. Age-adjusted mean value of duration of reactive hyperemia was significantly smaller in men with a smoking habit, diabetes mellitus, hypercholesterolemia or obesity, and in women with smoking habit, hypertension, diabetes mellitus or obesity. The number of risk factors significantly correlated with the duration of reactive hyperemia in both men (r=-0.56, p<0.001) and women (r=-0.62, p<0.001), suggesting that endothelial dysfunction increases with the number of risk conditions clustering in a single individual. CONCLUSIONS: Duration of reactive hyperemia reflects cardiovascular risk factors and decreases with the number of risk conditions. These findings suggest that the duration of reactive hyperemia measured with plethysmography is potentially useful for assessing endothelial dysfunction. PMID- 16377934 TI - Cutoff and target values for intra-abdominal fat area for prevention of metabolic disorders in pre- and post-menopausal obese women before and after weight reduction. AB - BACKGROUND: The Japan Society for the Study of Obesity originally proposed a cutoff value of >100 cm(2) for the intra-abdominal fat area (IFA) as a definition for "visceral fat obesity" in Japanese adults. There are no studies on the cutoff or target values after weight reduction in pre- and post-menopausal women. METHODS AND RESULTS: In the present study 149 pre-menopausal obese women (PreM, 43.3 years, 27.3 kg/m(2)) and 58 post-menopausal women (PostM, 53.9 years, 27.7 kg/m(2)) participated in a 14-week weight reduction program. The IFA was measured by computed tomography. The program induced significant reductions in body weight (8.6 kg in PreM and 7.8 kg in PostM). The IFA decreased significantly from 80.4 +/- 41.3 to 50.7 +/- 23.8 (PreM) and from 115.4 +/- 38.0 to 75.7 +/- 30.5 (PostM). CONCLUSIONS: The receiver-operating characteristic curve analyses revealed that the appropriate cutoff values were 80 cm(2) (PreM) and 110 cm(2) (PostM) before the program, and after the program the appropriate target values were determined as 60 and 70 cm(2), respectively. PMID- 16377936 TI - Effect of losartan and amlodipine on left ventricular diastolic function in patients with mild-to-moderate hypertension (J-ELAN): rationale and design. AB - BACKGROUND: Hypertension is a major underlying disease that may cause left ventricular (LV) diastolic dysfunction, even without LV systolic dysfunction, and antihypertensive drugs could affect LV diastolic function. METHODS AND RESULTS: The Effect of Losartan and Amlodipine on Left Ventricular Diastolic Function in Patients With Mild-to-Moderate Hypertension (J-ELAN) study is a multicenter, prospective, randomized trial designed to assess the effects of losartan and amlodipine on LV diastolic function in hypertensive patients with LV diastolic dysfunction in the absence of systolic dysfunction. A total of 300 patients (150 patients in each group) will be enrolled. In addition to Doppler echocardiographic indices of LV diastolic function, changes in LV structure and atherosclerosis of the carotid arteries will be serially assessed. The maximum follow-up period is 18 months. CONCLUSIONS: This study will provide the characteristic differences in the effects of amlodipine and losartan on LV diastolic dysfunction in hypertensive patients. PMID- 16377938 TI - Effects of folic acid and magnesium on the production of homocysteine-induced extracellular matrix metalloproteinase-2 in cultured rat vascular smooth muscle cells. AB - BACKGROUND: Hyperhomocysteinemia is an independent risk factor of coronary artery disease, but some studies have shown that patients with hyperhomocysteinemia are not prone to atherosclerosis. The aim of this study was to test whether homocysteine increases the production of matrix metalloproteinase-2 (MMP-2) and if extracellular additional magnesium and folic acid alters MMP-2 secretion. METHODS AND RESULTS: Gelatin zymography and western blotting were used to investigate the effects of different homocysteine levels (0-5,000 micromol/L) on MMP-2 production, and the effects of different folic acid concentrations (0-10 micromol/L) and magnesium concentrations (0-3.0 mmol/L) on homocysteine-induced MMP-2 in cultured rat vascular smooth muscle cells. Furthermore, the changes in MMP-2 were compared under various treatments for 24 h, 48 h and 72 h. Homocysteine (50-1,000 micromol/L) increased the production of MMP-2 significantly in a dose-dependent manner and at a high level (5,000 micromol/L) reduced the production of MMP-2. Increased production of MMP-2 induced by homocysteine was reduced by additional extracellular folic acid in a dose dependent manner. Magnesium also reduced the increase of MMP-2 production induced by homocysteine. Production of MMP-2 under various treatments for 72 h increased more than during 24 or 48 h. CONCLUSIONS: Homocysteine (50-1,000 micromol/L) significantly increased the production of MMP-2 in a dose-dependent manner. Added extracellular folic acid and magnesium decreased the homocysteine-induced MMP-2 secretion. These data suggest a beneficial effect of folic acid and magnesium on the pathogenesis of coronary artery disease. PMID- 16377937 TI - Mechanisms underlying nano-sized air-pollution-mediated progression of atherosclerosis: carbon black causes cytotoxic injury/inflammation and inhibits cell growth in vascular endothelial cells. AB - BACKGROUND: Epidemiological studies indicate a significant link between exposure to environmental air pollution and mortality and morbidity from ischemic heart disease. Because nanoparticles can translocate into blood circulation, the present study aimed to clarify their direct effects on human vascular endothelial cells (ECs). METHODS AND RESULTS: Human umbilical vein ECs (HUVECs) were treated with carbon black (CB), a component of diesel exhaust particles, for 24 h. CB induced cytotoxic morphological changes such as cytosolic vacuole formation, cell disorientation and decreased density. Lactate dehydrogenase assay revealed that CB induced cytotoxic injury in both the cells and plasma membranes. Proliferation assay showed that CB inhibited cell growth. Monocyte chemoattractant protein-1 but not vascular cell adhesion molecule-1 was induced by CB. CB reduced the expressions of connexin37 and endothelial nitric oxide (NO) synthase. Microarray analysis revealed the induction of pro-inflammatory molecules by CB. CONCLUSIONS: The present results demonstrate for the first time that CB directly affects the endothelium, causing cytotoxic injury, inflammatory responses, and inhibition of cell growth. As EC injury/inflammation and membrane disintegration are related to the initiation of atherosclerosis, and NO is anti-atherogenic and anti thrombogenic, the direct effects of nanoparticles on ECs may represent one mechanism behind environmental air pollution-mediated atherosclerosis and ischemic heart disease. PMID- 16377939 TI - Newborn infant with maternal anti-SSA antibody-induced complete heart block accompanying cardiomyopathy. AB - Newborn case of maternal anti-SSA antibody-induced congenital complete heart block (CCHB) accompanying cardiomyopathy is presented. Unexpectedly, she died of ventricular tachycardia, not bradycardia, 6 days after birth. Autopsy revealed left ventricular cardiomyopathy with endocardial fibroelastosis. Thus, when evaluating fetal cardiac performance in cases of maternal anti-SSA antibody induced CCHB, it is necessary to pay attention to myocardial attributes such as endocardial hyperplasia. PMID- 16377940 TI - The role of mammalian Staufen on mRNA traffic: a view from its nucleocytoplasmic shuttling function. AB - The localization of mRNA in neuronal dendrites plays a role in both locally and temporally regulated protein synthesis, which is required for certain forms of synaptic plasticity. RNA granules constitute a dendritic mRNA transport machinery in neurons, which move along microtubules. RNA granules contain densely packed clusters of ribosomes, but lack some factors that are required for translation, suggesting that they are translationally incompetent. Recently some of the components of RNA granules have been identified, and their functions are in the process of being examined, in attempts to better understand the properties of RNA granules. Mammalian Staufen, a double-stranded RNA binding protein, is a component of RNA granules. Staufen is localized in the somatodendritic domain of neurons, and plays an important role in dendritic mRNA targeting. Recently, one of the mammalian homologs of Staufen, Staufen2 (Stau2), was shown to shuttle between the nucleus and the cytoplasm. This finding suggests the possibility that Stau2 binds RNA in the nucleus and that this ribonucleoprotein particle is transported from the nucleus to RNA granules in the cytoplasm. A closer study of this process might provide a clue to the mechanism by which RNA granules are formed. PMID- 16377941 TI - Plasma leptin levels of elite endurance runners after heavy endurance training. AB - A decrease in testosterone levels and an increase in cortisol levels are observed in male athletes with the overtraining syndrome (OTS). Cortisol causes blood leptin levels to rise and testosterone has an inverse relationship with blood leptin levels. Therefore, we hypothesized that the hormonal changes as a result of OTS induce an increase in leptin. To test this hypothesis, we examined the relationship among changes in leptin, testosterone and cortisol in thirteen male collegiate distance runners (aged 20.3+/-1.1 years) before and after an 8-day strenuous training camp. Runners ran 284.1+/-48.2 km during the training camp. Body fat percentages and plasma glucose concentrations decreased significantly after the training. Non-ester fatty acids and total cholesterol concentrations in blood were unchanged. Serum cortisol concentrations showed a significant increase after the training camp (from 11.82+/-2.00 microg/dl to 16.78+/-3.99 microg/dl), and serum testosterone decreased significantly (from 408.0+/-127.6 ng/dl to 265.2+/-97.6 ng/dl). The ratio of testosterone to cortisol (TCR) dropped by 50% after training (from 35.62+/-13.69 to 16.94+/-8.47). These results suggest that the subjects reached a state of the OTS. Contrary to our hypothesis, plasma leptin was not significantly changed (from 1.34+/-0.29 ng/ml to 1.49+/-0.18 ng/ml). Delta Plasma leptin was not significantly correlated with delta serum cortisol, delta TCR or delta fat percentage. However, delta serum testosterone was positively correlated with delta plasma leptin (r=596, p<0.05). Plasma leptin concentrations might modulate the secretion of testosterone in overtraining conditions. In conclusion, the change in blood leptin level is independent of the changes in cortisol, TCR and fat percentage in highly trained male athletes in the state of the OTS. PMID- 16377942 TI - Relationships of anthropometrical parameters and body composition with bone mineral content or density in young women with different levels of physical activity. AB - The aim of the study was to test the possible relationships of anthropometrical parameters, somatotype and body composition parameters with bone mineral content (BMC) and bone mineral density (BMD, total body, the dominant arm distal radius, antero-posterior lumbar spine--L2-L4, femoral neck) in strength- (n=33) and endurance- (n=32) trained and sedentary normal-weight (n=41) and overweight (n=23) young females. Their body height and mass were measured and BMI calculated. Nine skinfolds, thirteen girths, eight lengths and eight breadths/lengths were measured. Somatotype components were calculated according to Carter and Heath (1990). Whole body fat percentage, fat mass, lean body mass (LBM), BMC and BMD were measured by DXA. The relationship of different BMC and BMD values at each of the regions studied to the different anthropometrical and body composition parameters were analysed by using a stepwise multiple regression analysis. In all groups, BMC is highly dependent on the body mass (31.5-81.2%, R2x100). In the endurance-trained females, BMD is dependent on LBM, especially in both weight-bearing sites (66.2% in L2-L4 and 35.3% in the femoral neck). LBM explained 77.0% of the total variance of BMC in this group. BMC in the strength trained group is dependent on the lower body anthropometrical parameters--thigh skinfold (18.2%), calf girth (25.2%), trochanterion length (24.1%) and sitting height (51.4%). From the endurance-trained group, BMC is dependent on hip girth (75.2%) or in combination with ankle girth (81.2%). From the length parameters, trochanterion is the most important (55.8%) and from breadths/lengths, sitting height (57.1%). In the normal-weight females, BMC is dependent on the calf girth (31.1%), trochanterion length (28.2%) and sitting height (29.8%). In the overweight group, only chest girth (20.1%) and biacromial breadth/length (27.0%) had a relationship with BMC. From somatotype components, only ectomorphy explained BMD in the endurance-trained females in the femoral neck (21.3%) and in the lumbar spine (20.9%). We can conclude that from the body composition parameters, LBM is a powerful predictor of BMC and BMD. From the anthropometrical parameters measured, lower body parameters are the most important. Somatotype components (ectomorphy) had a relationship with BMD only in the endurance-trained group. There are some differences that depend on the specific physical activity field. In the endurance-trained group, the anthropometry is more important than in the strength-trained group. PMID- 16377943 TI - The practice effect and its difference of the pursuit rotor test with the dominant and non-dominant hands. AB - The purpose of this study was to examine the practice effect and what difference it makes in the pursuit rotor test for the dominant and non-dominant hands in 30 right-handed Japanese male adults aged 18 to 23 years (Age 20.8+/-1.4 yrs). The subjects performed the pursuit rotor test for 1 min in 20 trials with a 1-min interval alternately using the dominant and non-dominant hands. After continuing for 10 trials, a 5-min rest was taken. The measurement order was randomly assigned. Contact time of a steel pen and a target was measured in units of 1/10 sec. The measurements showed a constant increasing tendency at every trial until the 6th trial in both hands. Significant linear regressions were identified, but the increase-rate of the dominant hand was significantly larger. Individual differences showed a decreasing tendency at every trial in the dominant hand, but in the non-dominant hand it increased until the 4th trial then decreased. The relationships between measurements of the 1st and 10th trials in both hands and both hands in the 1st or 10th trial were not high. The performance of the pursuit rotor test improves at every trial in both hands, but the improvement rate decreases after the 7th trial. The improvement rate of the dominant hand is high. The change in individual differences differs in both hands and the relationship between the measurements is not high. It can be judged that the practice effect of the pursuit rotor test differs in the dominant and non-dominant hands. PMID- 16377944 TI - Effects of chronic NH4Cl dosage and swimming exercise on bone metabolic turnover in rats. AB - To determine the effects of ammonium chloride (NH4Cl) dosage and swimming exercise training during 4 weeks on bone metabolic turnover in rats, seven-week old female 24 Wister-Kyoto (WKY) rats were investigated by bone status including bone mineral density (BMD) and biomechanical markers from blood and urine. Twenty four rats (initial weight: 191.2+/-7.6 g) were randomly divided into four groups: baseline (8 weeks old) control group (n=6, BC), 4-week control group (n=6, Con), 4-week swimming exercise loading group (n=6, Swim) and 4-week chronic NH4Cl dosage group (n=6, Acid). All rats were fed an AIN93M diet (Ca: 0.5%, P: 0.3%), and both Con and Swim groups were pair-fed by feeding volume of the NH4Cl dosage group. The acid group only received 0.25 M NH4Cl distilled water ad libitum. At the end of the experimental period, rats were sacrificed with blood drawn and femur and tibia were removed for analysis of bone mineral density (BMD) by dual energy X-ray absorptiometry (DEXA). In the Swim group, 24-hour urinary deoxypiridinoline (Dpd) excretion, reflecting bone resorption, was significantly increased (p<0.05) with a tendency towards decrease of BMD (N.S.), and body weight and abdominal fat weight were decreased in approximately 7% (p<0.05) and 58% (p<0.001), as compared with age matched Con rats. In the Acid group, 24-hour urinary calcium (Ca) and phosphorus (P) excretion were increased approximately 2.1-fold (p<0.05) and 2.0-fold (p<0.01), respectively, with increase of kidney weight as much as in the Con groups. Serum Ca and P concentration, as well as urinary Dpd excretion were, however, not significantly changed. These results suggest that blood Ca and P concentrations in the chronic acidosis condition during the 4-weeks might be maintained by hypercalciuria and hyperphosphaturia with kidney disorder, and swimming exercise training leads to decrease in BMD with stimulation of bone resorption and reduction of body fat. PMID- 16377945 TI - Specific physiological responses in women with severe primary dysmenorrhea during the menstrual cycle. AB - This study examined the specific physiological responses of women with primary dysmenorrhea during the severely painful menstrual (days 1-2 of menstruation) and the non-painful follicular phases (days 5-8 after the onset of menstruation). Subjects consisted of 10 severe primary dysmenorrheic (Group P) and 10 non dysmenorrheic women (Group C) with regular menstrual cycles. However, only 9 out of 10 and 8 out of 10 subjects of Groups P and C participated during the follicular phase. Physiological measures were taken in a resting state for 60 min. In the menstrual phase, the pain ratings and secretory immunoglobulin A (s IgA) concentrations of Group P were significantly higher than those of Group C, with relatively significant decreases in the leg-skin temperature in the former as well. In addition, the systolic (SBP) and diastolic blood pressure (DBP) at 45 min after rest in Group P were significantly higher than those found in Group C. These reactions strongly suggest activation of the sympathetic-adrenal-medullary axis (SAM axis) by painful stress. Furthermore, the low-frequency (LF) component of the SBP variability (SBPV) was significantly higher in Group P than Group C, even during the follicular phase. These findings imply that Group P may well have elevated activities of the SAM axis throughout the whole menstrual cycle. As such, it suggests that dysmenorrheic women may be affected by certain stressors other than pain per se and pain-derived emotions throughout the whole menstrual cycle. The findings also indicate that women with dysmenorrhea have more sensitive responses to the SAM system than non-dysmenorrheic women during stress. Moreover, the high-frequency (HF) component of heart rate variability (HRV), or the index for the vagus nerve activity, displayed a consistently higher value in Group P than C. It is postulated that the human body may have responded to pain in an attempt to maintain the homeostatic state by enhancing vagus nerve activity. PMID- 16377946 TI - Hematological change in venous blood of the lower leg during prolonged sitting in a low humidity and hypobaric environment. AB - The present study examined the effects of low humidity and hypobaric conditions on hematological change in venous blood of the lower leg during quiet prolonged sitting. Ten healthy male students participated as the subjects after singing a consent form to participate in this study. Their diet and water intake were controlled from 19:00 on the day before the experiments. The subjects sat for 130 min in a climatic chamber. Four experimental conditions in the chamber were designed from a combination of relative humidity (20% or 60%) and air pressure (sea level or equivalent to an altitude of 2,000 m). Ambient temperature was maintained at 24 degrees C in every condition. Venous blood was sampled from the lower leg before and after exposure to the experimental conditions, and was analyzed for blood viscosity and hematological indices. Also, body weight and leg circumference were measured as indices of total water loss and edema, respectively. Regarding the results of ANOVA, significant interactions between humidity and time were observed in blood viscosity, red blood cell count and hematocrit (each p<0.05). However, there were no significant differences in these indices among the conditions. Significant increases were observed in leg circumference (p<0.01), platelet count (p<0.05) and total protein (p<0.05) after the exposure compared with those before the exposure. There were no noticeable effects of hypobaric conditions in every measurement. In conclusion, prolonged sitting seems to be a more hazardous factor for thrombogenesis low humidity and hypobaric conditions during a long-distance flight. PMID- 16377947 TI - Topographic anatomy of the fornix as a guide for the transcallosal-interforniceal approach with a special emphasis on sex differences. AB - The topographic anatomy and morphometry of the fornix is important for standardizing the transcallosal-interforniceal approach and avoiding memory disturbances. The detailed morphometry of the fornix was investigated with a special emphasis on sex differences using midsagittal magnetic resonance imaging of 80 males and 102 females. Various parameters of the fornix, including the length of the upper and lower fornices, the curvature of the upper and lower fornices, and the insertion point of the fornix to corpus callosum, were investigated. The thickness of the fornix at the attachment point to the anterior commissure, the maximum distance to the upper and lower surfaces of the fornix, and the curvature of the upper and lower fornices showed sex differences (p < 0.5). The upper insertion point of the fornix to the corpus callosum was more frontal in females, but the functional relevance of these differences need further investigation. PMID- 16377948 TI - Recovery process of immediate prolonged posttraumatic coma following severe head injury without mass lesions. AB - The recovery process of immediate posttraumatic coma was investigated in 24 patients with severe head injury. The correlation between poor outcome in the recovery process and magnetic resonance (MR) imaging findings was analyzed. MR imaging was performed within the first 7 days for all patients. The recovery process was classified into phase 1 for recovery to moderately disabled and phase 2 to good recovery (GR) according to the Glasgow Outcome Scale. The median of phase 1 was 21.0 days. Four patients did not recover to GR and had poor outcome. Twenty patients recovered to GR. Thirteen patients had short phase 2 of under 10 days and seven patients had long phase 2 of over 60 days. All patients had abnormal lesions on MR imaging considered to be diffuse axonal injury. The number of lesions ranged from two to 10, with a mean of five. Lesions in the dorsal upper brainstem were significantly associated with poor outcome (p < 0.05). The combination of focal lesions in the callosal splenium and dorsal upper brainstem was most common in patients with poor outcome. Patients with long phase 2 had significantly more lesions than patients with short phase 2. PMID- 16377949 TI - Effect of platelet-activating factor receptor antagonist, etizolam, on resolution of chronic subdural hematoma--a prospective study to investigate use as conservative therapy. AB - Inflammatory reaction is very important for formation of the neomembrane of chronic subdural hematoma (CSDH). The present study evaluated medical treatment with the platelet-activating factor receptor antagonist, etizolam, for the resolution of CSDH, and the factors indicating surgery or conservative therapy. Alternate patients were assigned to the etizolam group or control group without medical treatment. Patients in the etizolam group received 3.0 mg etizolam per day for 14 days. A total of 53 patients were followed up for at least 6 months. Univariate analysis of differences in demographic characteristics, clinical findings, and initial computed tomography (CT) findings, and multiple logistic regression analysis of the relationship between etizolam treatment and requirement for surgery using age, sex, low density of hematoma on CT, and paresis as confounders were performed. Etizolam treatment (adjusted odds ratio [OR] 0.156, 95% confidence interval [CI] 0.024-0.999, p = 0.049) was negatively correlated with requirement for surgery. Low density of hematoma (adjusted OR 0.125, 95% CI 0.019-0.846, p = 0.033) was found to be an independent negative predictor, and paresis as an initial symptom (adjusted OR 6.35, 95% CI 1.04-38.7, p = 0.045) was an independent positive predictor of requirement for surgery. Etizolam administration can promote the resolution of CSDH, especially at the stage of hygroma appearing as low density on CT. Surgery is recommended if the patient presents with paresis. PMID- 16377950 TI - Acute subdural hematoma after intra-arterial thrombolysis for acute ischemic stroke--case report. AB - A 79-year-old man with a cardiac pacemaker for bradycardia fell down and presented with sudden onset of right hemiplegia and aphasia. Initial computed tomography (CT) showed no cerebral infarction but angiography revealed occlusion of the left middle cerebral artery (MCA). Local intra-arterial thrombolysis with tissue plasminogen activator (tPA; tisokinase, 1,600,000 units) was performed 3 hours after the onset, and the MCA was partially recanalized. Further administration of tPA was suspended because of nosebleed. However, the patient's neurological findings did not improve. His consciousness gradually deteriorated to coma and quadriplegia with dilation of the left pupil 2.5 hours after thrombolysis. CT disclosed marked mass effect with a left acute subdural hematoma and a small intracerebral hematoma in the left frontal lobe. He underwent urgent craniotomy and removal of the subdural hematoma. The subdural hematoma originated in a frontal cerebral contusion. He died of severe brain edema 2 days after surgery. Acute subdural hematoma is a very rare complication of intra-arterial thrombolysis. Presumably he had suffered head trauma at the first onset. Evidence of head trauma should be considered a contraindication for the use of thrombolytic agents in a patient with acute stroke. PMID- 16377951 TI - Subarachnoid hemorrhage associated with von Willebrand's disease--case report. AB - A 59-year-old woman with type IIA von Willebrand's disease (VWD) presented with subarachnoid hemorrhage (SAH). Computed tomography showed SAH in the right sylvian fissure and intracranial hemorrhage in the right temporal lobe. Angiography demonstrated an aneurysm at the bifurcation of the right middle cerebral artery. Neck clipping was performed on the 3rd day after the onset with intra- and postoperative administration of factor VIII/von Willebrand factor concentrate. No excessive bleeding occurred. Patients with prolonged bleeding time should be screened for VWD before surgery. This is a rare case of VWD presenting with SAH secondary to ruptured intracranial aneurysm. The clinical characteristics and the management of SAH in a patient with VWD are discussed. PMID- 16377952 TI - Mechanical thrombolysis for treatment of acute sinus thrombosis--case report. AB - A 55-year-old woman presented with consciousness disorders. Computed tomography revealed hemorrhage in the left temporoparietal region. The angiographic diagnosis was progressive sinus thrombosis from the superior sagittal sinus to the bilateral transverse sinuses. Her condition deteriorated despite heparin administration. Therefore, mechanical thrombolysis was performed for sinus thrombosis using a balloon catheter, in addition to supportive thrombolytic therapy with urokinase, resulting in sinus patency. Mechanical thrombolysis is an effective therapeutic modality for sinus thrombosis refractory to heparin administration. PMID- 16377953 TI - Subdural hematoma in a patient with Hageman trait--case report. AB - A 61-year-old man with severe factor XII deficiency presented with a subdural hematoma appearing as mixed but mainly high density by computed tomography in the left frontotemporoparietal region. No cranial injury was reported in the medical history of the patient. Clotting system study showed less than 1% functional activity of factor XII, whereas the levels of the other clotting factors were within the normal ranges. Partially clotted and hemolyzed subdural hematoma was removed through a craniotomy. The postoperative course was uneventful. The patient later died of severe circulatory-respiratory failure. We believe that the subdural hematoma may have developed as a result of a minor head trauma sustained in the past. We suggest that impairment of fibrinolytic activation related to severe factor XII deficiency might have contributed to the delay of dissolution of the subdural hematoma which, under ordinary circumstances, would have formed chronic subdural hematoma. PMID- 16377954 TI - Simultaneous cranial and spinal subdural hematoma. AB - A 59-year-old male presented with spinal subdural hematoma (SDH) with concomitant cranial chronic SDH manifesting as mild paraparesis and numbness in both lower extremities. Magnetic resonance (MR) imaging showed simultaneous occurrence of cranial and spinal SDHs. The patient was treated conservatively because of poor medical condition and mild neurological symptoms, and recovered well within 1 month. Serial follow-up MR imaging revealed spontaneous resolution of both lesions, with signal intensity changes suggesting the degenerative process of subacute hematoma. The spinal hematoma may have migrated from the cranial lesion. Spinal SDH is a potential sequela of chronic SDH in the cranium. PMID- 16377955 TI - Effect of oral colestimide on the elimination of high-dose methotrexate in patients with primary central nervous system lymphoma--case report. AB - Delayed methotrexate (MTX) elimination occurred in two patients with primary central nervous system lymphoma undergoing high-dose MTX treatment. Oral administration of the anion exchange resin colestimide, which binds MTX effectively in vitro, effectively accelerated MTX elimination. Colestimide probably interrupts the enterohepatic circulation, and is a potential oral antidote to MTX toxicity. PMID- 16377956 TI - Cyst of the ligamentum flavum--case report. AB - An 86-year-old man presented with intermittent claudication caused by a cyst of the ligamentum flavum. Lumbar magnetic resonance imaging demonstrated an extradural cystic mass at the L5-S1 intervertebral space and canal stenosis at the L4-5 space. L-5 laminectomy and flavectomy at the L4-5 and L5-S1 spaces were performed, and the cystic mass was excised. The histological features were consistent with cyst of the ligamentum flavum. The histological diagnosis was ganglion cyst of the ligamentum flavum. After surgery, claudication completely disappeared and the patient made a good recovery. PMID- 16377957 TI - Care for dying patients with primary malignant brain tumor. AB - Patients with primary malignant brain tumor experience deterioration of multi focal neurological deficits such as hemiparesis, aphasia, visual field defects, dysphagia, and disturbance of recent memory at the advanced stage of disease. With these advancing neurological deficits, many patients will inevitably prepare for death and may experience psychological and spiritual distress. Active listening is an important skill to explore the fears of patients with a terminal illness but in the advanced stage of a primary brain tumor, patients usually have great difficulty with verbal expression. Even if patients do not suffer from complete expressive aphasia, they often have difficulty verbalizing their thoughts and feelings. Sadly, disturbance of vocal expression is a common accompaniment of this pathology. Unless the pathophysiology is understood, an observer may fail to comprehend the patient's non-verbal communication. Seeking to understand these issues is a prerequisite of the preservation of dignity and provision of ethical care for such patients. PMID- 16377958 TI - N-methyl-D-aspartate and group I metabotropic glutamate receptors are involved in the expression of ethanol-induced sensitization in mice. AB - Effects of acamprosate and ionotropic uncompetitive N-methyl-D-aspartate receptor antagonists and group I metabotropic glutamatergic receptor antagonists on the expression of ethanol-induced sensitization were investigated in mice. The results indicated that acamprosate (200 and 400 mg/kg) and N-methyl-D-aspartate receptor antagonists, neramexane (10 and 20 mg/kg) and MK-801 (0.1 and 0.2 mg/kg), inhibited the expression of ethanol-induced sensitization. Acamprosate, but not the other compounds tested, also blocked the stimulant effect of acute injections of ethanol. Among the group I metabotropic glutamatergic receptor antagonists, only the metabotropic glutamatergic receptor 5 antagonist, MTEP (5, 10, and 20 mg/kg) showed an effect similar to the N-methyl-D-aspartate receptor antagonists. The metabotropic glutamatergic receptor 1 antagonist, EMQMCM (5, 10, and 20 mg/kg), however, potentiated the inhibitory effect of MK-801 on the expression of ethanol-induced sensitization. The findings indicate that glutamatergic neurotransmission is important in the ethanol-induced sensitization process, and suggest that co-administration of metabotropic glutamatergic receptor 1 antagonists and N-methyl-D-aspartate receptor antagonists may be useful in therapy for alcoholism. PMID- 16377959 TI - Anxiolytic-like activity of agomelatine and melatonin in three animal models of anxiety. AB - The activity of the novel antidepressant agomelatine was evaluated in three models of anxiety and compared with that of melatonin and two anxiolytics, diazepam and buspirone. All drugs were tested 2 h before and 2 h after the dark phase of the diurnal cycle. Morning and evening agomelatine (10-75 mg/kg) administration increased animals' responses in the elevated plus maze and Vogel tests. Melatonin (10-75 mg/kg) enhanced open arms exploration in the evening experiment and was inactive in the Vogel test. In the conditioned ultrasonic vocalization test, agomelatine, but not melatonin, was active in the morning and evening experiment. Melatonin antagonist, S22153 (20 mg/kg), enhanced the action of morning and evening agomelatine administration in the Vogel and conditioned ultrasonic vocalization tests, while in the elevated plus maze test, S22153 inhibited effects of evening but not morning melatonin and agomelatine administration. These results indicate the involvement of both the melatonin and the 5-HT2C receptors in the mechanism of anxiolytic-like action of agomelatine. PMID- 16377960 TI - Citalopram counteracts depressive-like symptoms evoked by chronic social stress in rats. AB - Recently, we have described a new model of chronic social stress in rats, based on the resident-intruder paradigm. In this model, rats show behavioural changes that may be considered correlates of depressive symptoms, such as anhedonia and motivational deficits. The present study was designed for pharmacological validation of this model. Animals were socially stressed for 5 weeks and, in parallel, after the first week of stress, they were subjected to chronic (4 weeks) treatment with the antidepressant drug citalopram. The drug was administered via drinking water (30 mg/kg). The optimal dose of citalopram was determined in a pilot study. After 4 weeks of treatment, plasma levels of citalopram and its metabolite were found to be within the human therapeutic range. The effects of social stress and citalopram treatment were assessed by behavioural tests. Chronically stressed rats showed reduced locomotor and exploratory activity, reduced sucrose preference and increased immobility time in the forced swimming test. Chronic oral administration of citalopram abolished those effects and normalized behaviours related to motivation and reward sensitivity. These observations provide evidence for the predictive validity of the chronic social stress paradigm as a model of depressive symptoms in rats. PMID- 16377961 TI - Use of the elevated plus-maze test with opaque or transparent walls in the detection of mouse strain differences and the anxiolytic effects of diazepam. AB - The elevated plus-maze is a widely employed behavioural assay for anxiolytic and anxiogenic agents in rodents. Among laboratories, however, the apparatus often differs between the use of transparent and opaque walls. This inconsistency may represent an unnecessary confound in the literature when comparing results. Here, we directly contrasted the two designs with respect to (1) mouse strain differences (C57BL/6, 129/Sv, and C3H/He) and (2) sensitivity to diazepam (0.5, 1.0, and 1.5 mg/kg, intraperitoneal). Both mazes yielded similar results indicating that 129/Sv mice were more anxious than C57BL/6 or C3H mice, with the transparent elevated plus-maze tending to encourage open-arm exploration in all three strains. Next, we examined the effect of diazepam in the 129/Sv strain across the two mazes. Systemic diazepam at 1.5 mg/kg led to increased percentage time spent in the open arms in both elevated plus-mazes; the drug was ineffective in both elevated plus-mazes at a dose of 0.5 mg/kg. Although our results revealed little practical difference between the two mazes in terms of their ability to detect differences in anxiety-related behaviour, the baseline difference in open arm exploration between the two elevated plus-mazes suggests that the transparent design may be more congenial for the detection of anxiogenic manipulations, and the opaque maze for anxiolytic manipulations. PMID- 16377962 TI - Level of response suppression and amphetamine effects on negatively punished adjunctive licking. AB - The purpose of the present study was to investigate whether the level of response suppression is a major determinant of the effect of D-amphetamine on negatively punished adjunctive drinking. Rats were initially exposed to a multiple fixed time (FT) 30-s FT 30-s food delivery schedule. They were then divided into two groups and subjected to one of two different multiple schedules, FT 30-s FT 45-s or FT 30-s FT 90-s. The FT 45-s and FT 90-s components were signalled by a tone. Comparably high levels of adjunctive licking were observed in both FT 30-s components, intermediate licking levels in the FT 45-s component, and little licking in the FT 90-s component. Licking during the FT 30-s components was subsequently punished by lick-contingent signalled delays (by a blackout) in food delivery. The duration of such delays was adjusted to reduce licking to levels obtained in the FT 45-s or FT 90-s components, respectively for each of the two groups. Punished licking was increased by 0.3 and 1.0 mg/kg of D-amphetamine, an effect that was greater in the FT 30-s FT 90-s group. No increase in licking was observed in the FT 45-s component, but the 1.0 mg/kg dose also increased responding in the FT 90-s component. In general, no statistically significant differences were found between the effects of D-amphetamine on punished and unpunished schedule-induced licking. As licking decreased during the FT 90-s component when the punishment contingency was introduced in the alternate component, the punishment procedure and FT 30-s component were entirely removed. On this occasion, D-amphetamine failed to increase licking induced by the FT 90-s schedule. These results indicate that the level of response suppression might be a good indicator of the degree to which D-amphetamine shows antipunishment effects on adjunctive licking reduced by negative punishment procedures. PMID- 16377963 TI - Effects of 5-HT2A receptor stimulation on the discrimination of durations by rats. AB - We recently found that rats' ability to discriminate durations of exteroceptive stimuli is disrupted by the non-selective 5-HT receptor agonist quipazine. Ketanserin reversed this effect, suggesting that the effect may be mediated by 5 HT2A receptors. Here, we report that the 5-HT2A/2C receptor agonist 2,5-dimethoxy 4-iodoamphetamine (DOI) also disrupts temporal discrimination, and that this effect can be reversed by ketanserin and the highly selective 5-HT2A receptor antagonist (+/-)2,3-dimethoxyphenyl-1-[2-(4-piperidine)-methanol] (MDL-100907). Twenty rats were trained to discriminate durations in a discrete-trials psychophysical procedure. In each 50-s trial, a light was presented for t seconds, following which two levers (A and B) were presented. A response on A was reinforced if t < 25 s, and a response on B if t > 25 s. Logistic psychometric curves were fitted to the proportional choice of B (%B) for derivation of timing indices [T50: time corresponding to %B = 50; Weber fraction: (T75-T25)/2T50, where T75 and T25 are times corresponding to %B = 75 and 25, respectively]. DOI 0.25 mg kg (subcutaneous) significantly increased the Weber fraction and tended to increase T50. Ketanserin 2 mg kg (subcutaneous) did not alter either parameter, but completely antagonized the effects of DOI. Similarly, MDL-100907 0.5 and 1 mg kg (intraperitoneal) did not affect performance, but completely antagonized the effects of DOI. The results indicate that the mixed 5-HT2A/2C receptor agonist DOI disrupts temporal discrimination via stimulation of 5-HT2A receptors. PMID- 16377964 TI - Effects of dose and time on the ability of alcohol to prime social drinkers. AB - Interoceptive drug cues, through associations with the drug's reinforcing properties, may act as conditioned stimuli and elicit conditioned responses. For instance, a dose of alcohol, given to alcohol-experienced people, can lead to an enhancement of alcohol drinking, a phenomenon known as the priming effect. The present study aimed to investigate the alcohol priming effect in non-dependent social drinkers with respect to the dose of alcohol preload and the time of testing after preload. Fifteen social drinkers participated in five daily consecutive sessions. On days 1 and 2 (training sessions), participants consumed a 500 ml beverage of either 0.6 g/kg of alcohol or placebo (50 ml aliquots) presented in 10 colour-coded cups. During days 3, 4 and 5 (testing sessions), a preload of placebo, 0.3 or 0.6 g/kg of alcohol was given (in randomized sequence) in 10 opaque colourless cups. Thirty, 60 and 90 min following the preload, participants responded to an imagery script referring to the drinks sampled at training including a question on the number of aliquots participants would consume from each of the drinks if given the opportunity (hypothetical choice). Participants completed questionnaires evaluating mood and alcohol desires at baseline (before the beverages were given) and after the hypothetical choice. The hypothetical choice showed significant interactions between dose and time: the greatest number of alcohol aliquots were wanted 30 min following the 0.6 g/kg dose of alcohol preload. Ratings from the Desires for Alcohol Questionnaire also showed that alcohol desires peaked 30 min following the 0.6 g/kg of alcohol preload. These data support previous evidence that priming with alcohol can occur and indicate that dose of, and time after preload might affect the strength of, the priming effect for alcohol-related behaviours. PMID- 16377965 TI - Effects of biasing the location of stimulus presentation, and the muscarinic cholinergic receptor antagonist scopolamine, on performance of a 5-choice serial reaction time attention task in rats. AB - A major purpose of the present studies was to determine the effects of varying the relative frequency, or bias, of stimulus presentation at different locations in a five-choice serial reaction time attention task. Training sessions were conducted in which the stimulus duration was held constant at 2 s and, initially, stimuli were presented with equal probability above each of the levers. During testing sessions, however, stimulus duration was either 0.2 or 2 s, and the frequency of presentation was varied among no bias, a middle bias, a left bias or a right bias condition. The training conditions were then changed such that the frequency of presentation was always a left bias, and test sessions were again conducted with no, middle, left or right bias. In the presence of the 0.2-s, but not the 2.0-s, stimulus, the percentage of correct responding on each of the choice levers during test sessions varied directly with the bias conditions in that the percentage of correct responding was highest at the most frequently presented location and lowest at the least frequently presented location. In addition, scopolamine, but not metscopolamine, increased omissions while also increasing anticipatory responses in rats trained under left-bias conditions and tested under right-bias conditions. The present findings suggest that varying the frequency of stimulus location can provide additional measures of attention in the five-choice serial reaction time task. PMID- 16377966 TI - A comparison of the effects of psychotomimetics and anxiolytics on punished and unpunished responding maintained by fixed interval schedules of food reinforcement in the rat. AB - Characterization of anxiolytic drugs often employs conflict paradigms in which the drug effects on punished and unpunished responding can be compared. In this study, a fixed interval schedule generating a range of baseline response rates allowed comparison of the effects of anxiolytic drugs with those of psychotomimetic drugs on equivalent and differing rates of punished and unpunished responding. The first response made by the rat after a 40-s fixed interval elapsed resulted in food pellet delivery. In punished intervals, signalled by the illumination of stimulus lamps above each lever, a 0.6-mA shock was delivered after every 20th response, resulting in a lower rate of responding than that in the unpunished intervals. Three psychotomimetic agents, D amphetamine, MK801 and DOI were compared with the anxiolytics chlordiazepoxide, NS2710 and pregabalin. The three psychotomimetics preferentially increased rates of unpunished responding compared with those of punished responding. Chlordiazepoxide, NS2710 and, to a lesser extent, pregabalin increased rates of both unpunished and punished responding. In comparison studies, yohimbine also increased rates of both unpunished and punished responding whereas the antidepressant citalopram had no effect. In conclusion, stable baseline performance over many months allowed the direct comparison of several different drugs in the same subjects with no need to adjust shock levels or equate baseline response rates. The drugs had systematic and replicable effects in this procedure, which, in the case of amphetamine and chlordiazepoxide, were similar to those in other species, and psychotomimetic drugs could clearly be distinguished from anxiolytic drugs. The procedure, however, has limited value for characterizing novel anxiolytic agents as the examples used here increased punished and unpunished responding to the same extent, and were indistinguishable in that regard from the clinically anxiogenic agent, yohimbine. PMID- 16377967 TI - Nonsteroidal antiinflammatory drugs in tendinopathy: friend or foe. PMID- 16377968 TI - Joint laxity and isokinetic thigh muscle strength characteristics after anterior cruciate ligament reconstruction: bone patellar tendon bone versus quadrupled hamstring autografts. AB - OBJECTIVES: To analyze ligamentous stability and isokinetic strength of knee extensor and flexor muscles after anterior cruciate ligament reconstruction using 2 different autografts with identical fixation. DESIGN: This study implements a comparative case series design. SETTING: Schulthess Clinic, Switzerland. PATIENTS: A total of 153 consecutive patients (n = 87 bone-patellar tendon-bone [BPTB]/66 quadruple semitendinosus/gracilis tendons [QSGT] patients) who underwent BPTB and QSGT surgery were functionally assessed. INTERVENTION: All patients were functionally assessed (knee extension/flexion isokinetic strength and knee joint laxity) at a mean follow-up time of 11 months. MAIN OUTCOME MEASUREMENTS: Laxity was measured utilizing the Kneelax arthrometer, whereas isokinetic strength of the knee extensor/flexor muscle groups was measured using the Biodex Multi Joint System 2 (180 degrees/s and 300 degrees/s). RESULTS: The QSGT group had significantly greater knee joint laxity when compared with the BPTB group (P < 0.001). Furthermore, females in the QSGT group demonstrated the greatest knee joint laxity overall (P < 0.001). Additionally, a significant flexor torque deficiency was demonstrated in the QSGT group (P < 0.001); however, no differences in extensor torque were observed. CONCLUSIONS: The use of a BPTB autograft achieved better knee joint stability as well as greater knee flexor torque than the QSGT autograft. Both grafts exhibited a similar knee extensor torque deficit, indicating that only quadriceps strength is not graft-dependent. PMID- 16377969 TI - Intrinsic factors associated with exercise-related leg pain in collegiate cross country runners. AB - OBJECTIVES: This study (1) examined the occurrence of exercise-related leg pain (ERLP) in collegiate cross-country athletes and (2) compared structural factors in runners with and without ERLP history. STUDY DESIGN: Cross-sectional design comparing 2 groups of cross-country athletes (with and without ERLP history). SETTING: Intercollegiate athletics. PARTICIPANTS: A total of 63 athletes (30 males, 33 females) representing 2 cross-country teams consented to participate. MEASURES: Athlete examination included self-report of ERLP history, active ankle dorsiflexion with knee extended and flexed, navicular drop (ND), and 1st ray length. ND index was calculated by dividing ND by 1st ray length. Athletes on 1 team (n = 32) were followed through 1 season to identify incidence of ERLP over the course of 1 season. Independent t tests were used to compare structural measures of those with and without ERLP history, and chi2 was used to examine frequencies. RESULTS: Of the 63 athletes, 33 (52%) reported ERLP history; most reported medial leg pain and also bilateral symptoms. No differences in structural measures were found between those with and without ERLP history. Ten of 32 athletes (31%) reported the occurrence of ERLP during the fall season, 8 of whom had a history of ERLP. CONCLUSIONS: Exercise-related leg pain was a common complaint among these athletes, usually occurring bilaterally and located along the medial leg. Athletes with ERLP history did not have a greater foot pronation as measured by ND, nor was there limited ankle range of motion as compared with those without ERLP history. Further research is needed to identify factors associated with the development of ERLP. PMID- 16377970 TI - A new view on adduction-related groin pain. AB - OBJECTIVE: To evaluate the hypothesis that groin pain at isometric hip adduction may not be caused by adductor tendinitis. DESIGN: Symptoms and signs in a cross sectional analysis. SETTING: Multicenter primary care institutes. SUBJECTS: Athletes with pain in the groin(s), provoked by playing sports, with a duration of complaints for at least 1 month and pain provocation on isometric adduction of the hips. INTERVENTIONS: Data on medical history and symptoms were collected. Pain provocation tests and strength measurements were performed. A pelvic belt was used to investigate its influence on pain provocation and strength. MAIN OUTCOME MEASUREMENTS: Site of the pain, duration of the complaints, severity of the pain, hip adduction force, pain at isometric hip adduction, restriction to perform active straight leg raising, influence of a pelvic belt on pain and strength of isometric hip adduction and straight leg raising. RESULTS: Groin pain was bilateral in 41%; pain was also located at the posterior aspect of the pelvis in 32%; Active Straight Leg Raise (ASLR) test was positive in 39%. When tested with a pelvic belt, the weakness of ASLR improved in all with a positive ASLR, hip adduction force increased significantly in 39% and pain at forceful isometric hip adduction decreased in 68%. CONCLUSIONS: Groin pain at isometric hip adduction may not be caused by adductor tendinitis in a large proportion of athletes with adduction-related groin pain. The results suggest that adduction related groin pain with a positive belt test may be treated by stabilization of the pelvis. PMID- 16377971 TI - Survey of sport participation and sport injury in Calgary and area high schools. AB - OBJECTIVE: To examine (1) sport participation and (2) sport injury in adolescents. DESIGN, SETTING, AND PARTICIPANTS: This was a retrospective survey design. In total, 2873 adolescents were recruited from a random sample of classes from 24 Calgary and area high schools. Each subject completed an in-class questionnaire in March 2004. MAIN OUTCOME MEASUREMENTS: Overall and sport specific participation rates (number of sport participants/number of students completing survey). Overall and sport-specific injury rates (number of injuries/number of participants). RESULTS: In the previous 1 year, 94% of students participated in sport. The top 5 sports by participation for males were basketball, hockey, football, snowboarding, and soccer, and for females, basketball, dance, volleyball, snowboarding, and soccer. The injury rate including only injuries requiring medical attention was 40.2 injuries/100 adolescents/y (95% CI, 38.4-42.1), presenting to a hospital emergency department was 8.1 injuries/100 adolescents/y (95% CI, 7.1-9.2), resulting in time loss from sport was 49.9 injuries/100 adolescents/y (95% CI, 48-51.8), and resulting in loss of consciousness was 9.3 injuries/100 adolescents/y (95% CI, 8.3-10.5). The greatest proportion of injuries occurred in basketball, hockey, soccer, and snowboarding. The top 5 body parts injured were the ankle, knee, head, back, and wrist. The top 5 injury types were sprain, contusion, concussion, fracture, and muscle strain. A previous injury was associated with 49% of the injuries and direct contact with 45% of injuries. CONCLUSIONS: Rates of participation in sport and sport injury are high in adolescents. Future research should focus on prevention strategies in sports with high participation and injury rates to maximize population health impact. PMID- 16377972 TI - The use of dietary supplements and medications by Canadian athletes at the Atlanta and Sydney Olympic Games. AB - OBJECTIVE: To learn more about the prevalence of dietary supplement and medication use by Canadian athletes in the Olympic Games in Atlanta 1996 and Sydney 2000. SETTING AND PARTICIPANTS: Data were collected from personal interviews with Canadian athletes who participated at the 1996 Atlanta and 2000 Sydney Olympic Games. The athletes were interviewed by Canadian physicians regarding the use of vitamins, minerals, nutritional supplements, and prescribed and over-the-counter medications. Of the 271 Canadian athletes who participated at the Atlanta Olympics, 257 athletes were interviewed; at the Sydney Olympics, 300 of 304 Canadian athletes were interviewed. MAIN OUTCOME MEASUREMENT: A quantitative and qualitative description of the use of dietary supplements by Canadian athletes at the Atlanta and Sydney Olympics. RESULTS: At the Atlanta Games, 69% of the athletes used some form of dietary supplements, whereas 74% of the athletes used dietary supplements at the Sydney Games. Vitamins were taken by 59% of men and 66% of women in Atlanta, and 65% of men and 58% women in Sydney. Mineral supplements were used by 16% of men and 45% of women in Atlanta, and 30% of men and 21% of women in Sydney. Nutritional supplements were used by 35% of men and 43% of women in Atlanta, and 43% of men and 51% of women in Sydney. The most popular vitamins were multivitamins in both Olympics. The most popular mineral supplements were iron supplements. The most commonly used nutritional supplement in Atlanta was creatine (14%), but amino acids (15%) were the most commonly used nutritional supplement in Sydney. In Atlanta, 61% of the athletes were using some form of medication, 54% of the athletes were using medications in Sydney. Nonsteroidal antiinflammatory drugs (NSAIDS) were the most commonly used medications at both Olympic Games. Among all sports, the highest prevalence of vitamin use occurred in boxing (91%) in Atlanta and swimming (76%) in Sydney. Rowers (56%) and cyclists (73%) demonstrated the highest use of mineral supplements. Nutritional supplement use occurred most often in swimming (56%) and cycling (100%). The use of NSAIDs was highest in softball (60%) in Atlanta and gymnastics (100%) in Sydney. CONCLUSION: This review demonstrates that dietary supplement use was common among Canadian athletes at both the Atlanta and Sydney Olympic Games. There was a slight increase in total dietary supplement use at the Sydney Games. Widespread use of supplements, combined with an absence of evidence of their efficacy and a concern for the possibility of "inadvertent" doping, underscore the need for appropriately focused educational initiatives in this area. PMID- 16377973 TI - Prospective study of bacterial and viral contamination of exercise equipment. AB - OBJECTIVE: Transmission of bacterial and viral pathogens is known to occur by hand contact with fomites. Exercise equipment in public facilities may serve as such fomites. It is not known whether equipment disinfection might reduce microorganism colonization. We performed studies to address these issues. DESIGN: Observational study of bacterial and viral culture results from hand-contact surfaces of exercise equipment, pre-exercise and postexercise; prospective study of viral culture results before and after intervention with disinfection solution. SETTING: Two fitness centers in a military community. INTERVENTION: One week trial of twice-a-day equipment disinfection. MAIN OUTCOME MEASURES: Type and number of bacteria and type of viruses present on equipment before and after exercise; prevalence of viral culture positivity on equipment before and after intervention. RESULTS: Bacterial cultures of body contact surfaces on equipment revealed benign bacterial species (coagulase-negative staphylococci, diphtheroids, and so forth) but no pathogenic bacteria whether obtained pre exercise or postexercise, or whether from aerobic versus weight training equipment. Viral cultures revealed the presence of viruses (generally rhinoviruses) on 63 of 100 (63%) hand-contact surfaces of equipment. Weight equipment was significantly more often contaminated than aerobic equipment (73% vs. 51%; P = 0.026). Disinfection of equipment did not lower the prevalence of virus isolation (48% positive before cleaning; 86% positive after cleaning). CONCLUSIONS: There is little risk of exposure to pathogenic bacteria on exercise equipment. Such equipment may commonly serve as fomites for the transmission of viruses. These data do not suggest that disinfection of exercise equipment will offer significant protective effects against virus exposure. PMID- 16377974 TI - Evaluation of maximal exercise performance, fatigue, and depression in athletes with acquired chronic training intolerance. AB - OBJECTIVE: This study compared differences in maximal strength and aerobic capacity and symptoms of fatigue and depression in athletes with acquired training intolerance (ATI) and control athletes (CON) matched for age and current training volume who did not have symptoms of excessive or chronic fatigue associated with their sporting activity. SETTING: University of Cape Town, Sports Science Institute of South Africa. PARTICIPANTS: Twenty ATI and 10 CON athletes participated in the trial. Although the ATI athletes reported symptoms of excessive fatigue during exercise, or symptoms of fatigue that occurred at rest and during activities of daily living, they did not fulfill the criteria for a diagnosis of chronic fatigue syndrome. MAIN OUTCOME MEASURES: A training and comprehensive medical history was recorded from all subjects. The Beck Depression Inventory Short Form (BDI-SF) was used to assess levels of depression in both ATI and control subjects. Maximal force output during a 5-second isometric voluntary knee extensor muscle contraction, and maximal aerobic capacity (VO2max), maximal heart rate (HRmax), and maximal blood lactate concentrations during a treadmill running test were measured in all subjects. RESULTS: There were no differences in maximal isometric force output, peak treadmill running speed, VO2max, HRmax, or blood lactate concentration at rest or after maximal exercise testing between the ATI and CON athletes. However, the BDI-SF scores were higher in the ATI (7.7 +/- 6.6 arbitrary units) than in the CON athletes (1.7 +/- 1.5 arbitrary units; (P = 0.0052). CONCLUSIONS: These findings suggest that the symptoms of excessive fatigue and acquired training intolerance described by these ATI athletes do not affect their maximal isometric and maximal aerobic capacity, and may be associated with psychologic depression in these athletes. PMID- 16377975 TI - Effects of a controlled program of moderate physical exercise on insulin sensitivity in nonobese, nondiabetic subjects. AB - OBJECTIVE: To study the effect of a moderate, aerobic physical exercise program on insulin resistance and its accompanying metabolic changes in a group of healthy, middle-age, nonobese subjects, without modifying oxygen consumption and body weight. DESIGN: The inclusion of subjects was carried out among volunteers from the health personnel of our center, who complied with the inclusion criteria. PARTICIPANTS: Twelve subjects (age 30-60 years, 5 females), nonsmokers, body mass index (BMI) <27 kg/m2 and fasting plasma glucose <6.1 mmol/L. INTERVENTIONS: Insulin resistance was assessed using the Bergmann minimal model modified with insulin, and basal and maximum metabolic rate were measured with standard methods. All subjects completed a 2-month program of aerobic exercise using the American College of Sports Medicine guidelines, consisting of aerobic exercise of moderate, regular, and continual intensity (3 times per week), with a duration of 45 to 50 minutes per session. RESULTS: Following controlled exercise, no significant differences in BMI, waist-hip ratio, blood pressure, lipids, free fatty acids, and leptin plasma values were observed. Plasma glucose and insulin values decreased significantly (-0.37 mmol/L and -16.5 pmol/L, respectively). Insulin sensitivity showed an increase of 2.1 x 10(-4) mU L(-1) min(-1) (P = 0.001). Basal and maximum metabolic rate showed no significant differences after the exercise program. CONCLUSIONS: Moderate aerobic exercise increases insulin sensitivity in nonobese, nondiabetic subjects in the absence of significant changes in weight, BMI, waist-hip ratio, lipid profile, and oxygen consumption. PMID- 16377976 TI - Overweight children reduce their activity levels earlier in life than healthy weight children. AB - OBJECTIVE: To determine differences in the time overweight and nonoverweight youth spend on light, moderate, hard, and very hard activity plus gender and age differences in activity patterns. DESIGN AND SETTING: This study was a cross sectional design in a primary care hospital. PARTICIPANTS: A total of 197 children and adolescents (4-16 years) participated. The overweight group (n = 107) were on a waiting list for an obesity treatment program and were defined as overweight, with a body mass index >95th percentile, and the nonoverweight group (n = 90) were volunteers recruited via advertisements and had a body mass index <75th percentile. INTERVENTIONS AND OUTCOME MEASUREMENTS: To determine activity patterns and perception of activity level, an interview was conducted with the child and a parent using a validated 7-day recall. Body fat was measured with bioelectrical impedance. RESULTS: There was a negative correlation between adiposity and total amount of activity performed, with the amount of moderate activity the best predictor of obesity. Girls did less activity than boys whether overweight or not. Between the ages of 4 and 7 years, overweight and nonoverweight subjects did similar amounts of activity. In the older subjects, the activity of the overweight was less than in nonoverweight. The overweight subjects perceived themselves to be just as active as others their same age and sex. CONCLUSIONS: Clinicians need to be aware that overweight children and adolescents are less active than nonoverweight children at an earlier age, particularly females, but feel that their activity level is similar to that of nonobese children. PMID- 16377977 TI - Gender comparisons of dynamic restraint and motor skill in children. AB - OBJECTIVES: To assess differences in neuromuscular dynamic restraint between high skilled and low-skilled prepubescent girls and boys. To determine the contribution of sport experience and physical characteristics to motor skill. SUBJECTS: Nineteen girls and 17 boys (8.89-9.40 y) participated. INTERVENTION: Isometric hamstring and quadriceps muscle strength was assessed. Subjects performed 3 landing trials for measurement of preparatory EMG and vertical leg stiffness. Motor skill was assessed through analysis of 12 fundamental tasks. Sport experience was reported as hours per week and total years in organized and nonorganized activity. OUTCOME MEASURES: Dynamic restraint variables of isometric strength, preparatory EMG activity, and vertical leg stiffness were measured between groups. The contributions of time in sport, type of sport, and physical characteristics on skill were analyzed. RESULTS: No significant gender or skill differences were found in quadriceps strength (P = 0.73), hamstring strength (P = 0.96), hamstring-to-quadriceps ratio (P = 0.71), or vertical leg stiffness (P = 0.38). Low-skilled children exhibited significantly greater (47.8%) preparatory hamstring-quadriceps coactivation than high-skilled subjects (P = 0.03). Participation in organized and nonorganized sport accounted for 29% of the variance in motor skill. CONCLUSIONS: Neuromuscular differences between genders were not observed, but dynamic restraint EMG measures differed between skill levels. The factors predisposing females to noncontact injuries may develop prepuberty to postpuberty from a combination of variables. Greater coactivation in the low-skilled group appears consistent with immature feedforward neuromuscular control strategies. These unrefined motor skills are less economical, may compromise dynamic restraint, and appear partially determined by sport experience. PMID- 16377978 TI - Effects of creatine supplementation on cerebral white matter in competitive sportsmen. AB - OBJECTIVES: To determine the neurobiochemical sequelae of oral creatine monohydrate supplementation in active athletes. DESIGN/PARTICIPANTS: Eighteen sportsmen underwent single-voxel proton magnetic resonance spectroscopy of the deep frontal cerebral white matter before and after 5 days of oral ingestion: 12 of 18 swallowed 4 x 5 g creatine monohydrate per day, and the remaining swallowed a placebo. MAIN OUTCOME MEASUREMENTS: Creatine, choline, and N-acetyl spectral resonances were evaluated at both long (135 ms) and short (20 ms) echo times. RESULTS: A mixed-design factorial ANOVA demonstrated no interaction over time in any of the measures (P at least 0.081). CONCLUSIONS: The results suggest that, for the given dosage regimen, ingested creatine augmentation does not alter the magnetic resonance visible creatine pool in the deep frontal cerebral white matter of young active sportsmen. PMID- 16377979 TI - Ribose versus dextrose supplementation, association with rowing performance: a double-blind study. AB - OBJECTIVE: It has been hypothesized that ribose supplementation rapidly replenishes adenosine triphosphate stores and thereby improves exercise performance. We compared the effects of ribose versus dextrose on rowing performance. DESIGN: Double-blind randomized trial. SETTING: Rowing team training area of large midwestern university. PARTICIPANTS: Thirty-one women collegiate rowers. INTERVENTIONS: We studied the effects of ribose versus dextrose supplementation (10 g each in 8 oz water) for 8 weeks before and after practice and 2000-m time trials. OUTCOME MEASUREMENTS AND RESULTS: In the time trials, the dextrose group showed significantly more improvement at 8 weeks than the ribose group (median, 15.2 vs. 5.2 s; P = 0.031). CONCLUSIONS: We doubt ribose impaired, and hypothesize dextrose enhanced, rowing performance. Further research is needed to define what role, if any, dextrose and ribose play as athletic supplements. PMID- 16377980 TI - Moderate exercise-induced hyponatremia. PMID- 16377981 TI - Extensor digitorum longus tendonitis in windsurfing due to footstrap fixation. PMID- 16377982 TI - Bilateral carotid artery dissection caused by springboard diving. PMID- 16377983 TI - Paget-Schroetter syndrome in a collegiate football player. PMID- 16377984 TI - Piezogenic pedal papules in a marathon runner. PMID- 16377985 TI - Adolescent athlete's shoulder: a case series of proximal humeral epiphysiolysis in nonthrowing athletes. PMID- 16377986 TI - Does physical activity affect changes in weight and waist size in midlife women? PMID- 16377987 TI - Effectiveness of a single corticosteroid injection for chronic rotator cuff tendinosis. PMID- 16377988 TI - Intrinsic risk factors for inversion ankle sprains in physical education students. PMID- 16377989 TI - Comparison of 2 eccentric exercise protocols for patellar tendinopathy in volleyball players. PMID- 16377990 TI - Hearing aid patients in private practice and public health (Veterans Affairs) clinics: are they different? AB - OBJECTIVE: In hearing aid research, it is commonplace to combine data across subjects whose hearing aids were provided in different service delivery models. There is reason to question whether these types of patients are always similar enough to justify this practice. To explore this matter, this investigation evaluated similarities and differences in self-report data obtained from hearing aid patients derived from public health (Veterans Affairs, VA) and private practice (PP) settings. DESIGN: The study was a multisite, cross-sectional survey in which 230 hearing aid patients from VA and PP audiology clinic settings provided self-report data on a collection of questionnaires both before and after the hearing aid fitting. Subjects were all older adults with mild to moderately severe hearing loss. About half of them had previous experience wearing hearing aids. All subjects were fitted with wide-dynamic-range-compression instruments and received similar treatment protocols. RESULTS: Numerous statistically significant differences were observed between the VA and PP subject groups. Before the fitting, VA patients reported higher expectations from the hearing aids and more severe unaided problems compared with PP patients with similar audiograms. Three wks after the fitting, VA patients reported more satisfaction with their hearing aids. On some measures VA patients reported more benefit, but different measures of benefit did not give completely consistent results. Both groups reported using the hearing aids an average of approximately 8 hrs per day. VA patients reported age-normal physical and mental health, but PP patients tended to report better than typical health for their age group. CONCLUSIONS: These data indicate that hearing aid patients seen in the VA public health hearing services are systematically different in self-report domains from those seen in private practice services. It is therefore risky to casually combine data from these two types of subjects or to generalize research results from one group to the other. Further, compared with PP patients, VA patients consistently reported more favorable hearing aid fitting outcomes. Additional study is indicated to explore the determinants of this result and its generalizability to other public health service delivery systems such as those in other countries. Moreover, efforts should be made to assess the potential for transferring positive elements from the VA system to the PP service delivery system, if possible. PMID- 16377991 TI - Working in noise with a hearing loss: perceptions from workers, supervisors, and hearing conservation program managers. AB - OBJECTIVE: Workers with hearing loss face special problems, especially when working in noise. However, conventional hearing conservation practices do not distinguish between workers with normal hearing versus impaired hearing. This study collected information from workers with self-reported noise exposure and hearing loss, supervisors of such workers, and hearing conservation program managers through focus groups and in-depth interviews to evaluate their perspectives on the impact of hearing loss on safety and job performance, the use of hearing protection, and information needed to appropriately manage hearing impaired workers who work in noisy environments. RESULTS: Concerns about working in noise with a hearing loss could be grouped into the following 10 categories: impact on job performance, impact on job safety, impaired ability to hear warning signals, impaired ability to monitor equipment, interference with communication, stress and/or fatigue, impaired communication caused by hearing protector use, reduced ability to monitor the environment as the result of hearing protector use, concerns about future quality of life, and concerns about future employability. Mostly, there was an agreement between the perceptions of workers, supervisors, and hearing conservation program managers regarding difficulties associated with hearing loss and consequent needs. These findings suggest that noise-exposed workers with hearing loss face many of the same problems reported in the literature by noise-exposed workers with normal hearing, with additional concerns primarily about job safety as the result of a reduced ability to hear environmental sounds, warning signals, and so forth. CONCLUSIONS: The study outlines potential challenges regarding job safety and hearing conservation practices for noise-exposed, hearing-impaired workers. Awareness of these issues is a necessary first step toward providing appropriate protective measures for noise-exposed, hearing-impaired workers. PMID- 16377992 TI - Speech and voice physiology of children who are hard of hearing. AB - OBJECTIVE: The purpose of this investigation was to examine the nature and frequency of deviant speech and voice physiology in children who are hard of hearing (HH). DESIGN: Ten HH children (age, 5 to 15 yrs) participated. Their hearing losses ranged in severity from moderate to severe. The following speech/voice physiological measures were examined: frequency of occurrence of negative intraoral air pressure (-Po), magnitude of Po, phonatory air flow, nasal air flow, voice onset time (VOT), and fundamental frequency (F0). Findings were compared with data previously collected from 56 children with normal hearing and 7 children with cochlear implants (Higgins, McCleary, Carney, & Schulte, 2003). RESULTS: Five of the 10 HH children exhibited deviant speech/voice behaviors. Only one showed deviancy on more than one measure. In addition, 8 HH children had some borderline-deviant speech behavior. The frequency and degree of speech/voice deviancy for the children in the present study was far less than what we previously had observed for children who underwent cochlear implantation after 5 yrs of age (Higgins et al., 2003). This was the case even for a child with a cochlear implant from our earlier study who eventually achieved speech perception scores that were as good as or better than some of the HH children in the present investigation. CONCLUSIONS: Deviant speech/voice physiology occurs to a limited extent for some HH children. Of the measures that we examined, those related to vocal fold tension and vocal fold articulation appeared to be most sensitive to the effects of diminished auditory input and feedback that occurs for HH children. Data from this and other studies suggest that abnormally high F0 occurs for about 20% of HH children and may be associated with poor speech perception skills. Future studies should examine whether earlier implantation can help deaf children to produce speech that is at least as normal as that of HH children. PMID- 16377993 TI - Threshold prediction using the auditory steady-state response and the tone burst auditory brain stem response: a within-subject comparison. AB - OBJECTIVE: The purpose of this study was to evaluate the accuracy with which auditory steady-state response (ASSR) and tone burst auditory brain stem response (ABR) thresholds predict behavioral thresholds, using a within-subjects design. Because the spectra of the stimuli used to evoke the ABR and the ASSR differ, it was hypothesized that the predictive accuracy also would differ, particularly in subjects with steeply sloping hearing losses. DESIGN: ASSR and ABR thresholds were recorded in a group of 14 adults with normal hearing, 10 adults with flat, sensorineural hearing losses, and 10 adults with steeply sloping, high-frequency, sensorineural hearing losses. Evoked-potential thresholds were recorded at 1, 1.5, and 2 kHz and were compared with behavioral, pure-tone thresholds. The predictive accuracy of two ABR protocols was evaluated: Blackman-gated tone bursts and linear-gated tone bursts presented in a background of notched noise. Two ASSR stimulation protocols also were evaluated: 100% amplitude-modulated (AM) sinusoids and 100% AM plus 25% frequency-modulated (FM) sinusoids. RESULTS: The results suggested there was no difference in the accuracy with which either ABR protocol predicted behavioral threshold, nor was there any difference in the predictive accuracy of the two ASSR protocols. On average, ABR thresholds were recorded 3 dB closer to behavioral threshold than ASSR thresholds. However, in the subjects with the most steeply sloping hearing losses, ABR thresholds were recorded as much as 25 dB below behavioral threshold, whereas ASSR thresholds were never recorded more than 5 dB below behavioral threshold, which may reflect more spread of excitation for the ABR than for the ASSR. In contrast, the ASSR overestimated behavioral threshold in two subjects with normal hearing, where the ABR provided a more accurate prediction of behavioral threshold. CONCLUSIONS: Both the ABR and the ASSR provided reasonably accurate predictions of behavioral threshold across the three subject groups. There was no evidence that the predictive accuracy of the ABR evoked using Blackman-gated tone bursts differed from the predictive accuracy observed when linear-gated tone bursts were presented in conjunction with notched noise. Similarly, there was no evidence that the predictive accuracy of the AM ASSR differed from the AM/FM ASSR. In general, ABR thresholds were recorded at levels closer to behavioral threshold than the ASSR. For certain individuals with steeply sloping hearing losses, the ASSR may be a more accurate predictor of behavioral thresholds; however, the ABR may be a more appropriate choice when predicting behavioral thresholds in a population where the incidence of normal hearing is expected to be high. PMID- 16377994 TI - Clinical evaluation of the Clarion CII HiFocus 1 with and without positioner. AB - OBJECTIVE: To study the clinical outcomes concerning speech perception of the Clarion CII HiFocus 1 with and without a positioner and link those outcomes with the functional implications of perimodiolar electrode designs, focusing on intrascalar position, insertion depth, stimulation levels, and intracochlear conductivity pathways. DESIGN: The speech perception scores of 25 consecutive patients with the Clarion CII HiFocus 1 implanted with a positioner and 20 patients without a positioner were prospectively determined. Improved multislice CT imaging was used to study the position of the individual electrode contacts relative to the modiolus and their insertion depth. Furthermore, stimulation thresholds, maximum comfort levels, and dynamic ranges were obtained. Finally, these data were associated with intracochlear conductivity paths as calculated from the potential distribution acquired with electrical field imaging. RESULTS: Implantation with a Clarion Hifocus 1 with positioner showed significantly higher speech perception levels at 3 mos, 6 mos, and 1 yr (p < 0.05) after implantation. Basally, the positioner brought the electrode contacts significantly closer to the modiolus, whereas apically no difference in distance toward the modiolus was present. Moreover, the patients with the electrode array in a perimodiolar position showed deeper insertions. The T-levels and dynamic range were not significantly different between the positioner and nonpositioner patients. Furthermore, the intracochlear conductivity paths showed no significant differences. However, a basal current drain is present for the shallowly inserted nonpositioner patients. CONCLUSIONS: A basally perimodiolar electrode design benefits speech perception. The combination of decreased distance to the modiolus, improved insertion depth, and insulating properties of the electrode array have functional implications for the clinical outcomes of the perimodiolar electrode design. Further research is needed to elucidate their individual contributions to those outcomes. PMID- 16377995 TI - A validation and potential clinical application of multivariate analyses of distortion-product otoacoustic emission data. AB - OBJECTIVE: To test the generalizability of multivariate analyses of distortion product otoacoustic emission (DPOAE) data. Previously published multivariate solutions were applied to a new set of data to determine if test-performance improvements, evident in previous reports, are retained. An additional objective was to provide an alternative approach for making multivariate dichotomous decisions of hearing status in the clinic, based on DPOAE measurements. DESIGN: DPOAE level and noise were obtained in 345 ears of 187 subjects. Approximately one third of the subjects had normal hearing, whereas the remainder had hearing loss, ranging from 25 to more than 120 dB HL. DPOAE data were collected at each of nine frequencies. After data collection, clinical decision theory, in combination with univariate (DPOAE level and signal-to-noise ratio [SNR]) and multivariate (logistic regression) analyses, was used to construct relative operating characteristic (ROC) curves and to generate ROC curve areas. In addition, test performance was assessed by fixing the false-alarm rate and comparing different approaches to analyses in terms of their failure rates as a function of magnitude of hearing loss. The DPOAE test results were compared with either single-frequency or multifrequency gold standards. The multivariate solutions were taken from previously published work (Dorn et al., 1999; Gorga, et al., 1999). RESULTS: DPOAE level and SNR resulted in roughly equivalent test performance (ROC curve areas and failure rates among ears with hearing loss), although DPOAE level performed better for frequencies above 1 kHz, and SNR performed better for frequencies at 0.75 and 1 kHz. Multivariate analyses resulted in better test performance for nearly all conditions, compared with the univariate approaches that used either DPOAE level or SNR. The improvements in test performance were greatest for the frequencies at which the univariate analyses performed poorest (0.75 kHz, 1 kHz, and 8 kHz). Less difference was observed between univariate and multivariate approaches when multifrequency gold standards were used; however, even for the multifrequency cases, multivariate analyses generally resulted in better performance. An approach that might facilitate the interpretation of multifrequency DPOAE measurements in the clinic is described. CONCLUSIONS: Previously described multivariate analyses were robust in that they improved test performance when applied to an entirely new set of DPOAE data. This, in turn, suggests that the previously described multivariate solutions may have clinical utility in that they are expected to improve test performance at no additional cost in terms of data-acquisition or data-analysis time. In addition to demonstrating that these solutions generalized to new data, an alternative approach to interpreting multifrequency DPOAE measurements is provided that includes the advantages of using multivariate analyses. This new metric may be useful when DPOAEs are used for screening purposes. PMID- 16377996 TI - GIN (Gaps-In-Noise) test performance in subjects with confirmed central auditory nervous system involvement. AB - OBJECTIVE: The purpose of the present study was to investigate the value of a new gap detection procedure called Gaps-In-Noise (GIN) for assessment of temporal resolution in a clinical population. DESIGN: The test consists of 0 to 3 silent intervals ranging from 2 to 20 msec embedded in 6-sec segments of white noise. The location, number, and duration of the gaps per noise segment vary throughout the test for a total of 60 gaps presented in each of four lists. The GIN procedure was administered to 50 normal-hearing listeners (group I) and 18 subjects with confirmed neurological involvement of the central auditory nervous system (group II). RESULTS: Results showed mean approximated gap detection thresholds of 4.8 msec for the left ear and 4.9 msec for the right ear for group I. In comparison, results for group II demonstrated a statistically significant increase in gap detection thresholds, with approximated thresholds of 7.8 msec and 8.5 msec being noted for the left and right ears, respectively. Significant mean differences were also observed in the overall performance scores (i.e., the identification of the presence of the gaps within the noise segments) of the two groups of subjects. Finally, psychometric functions, although similar for short and long duration gaps, were highly different for gaps in the 4- to 10-msec range for the two groups. CONCLUSIONS: A variety of psychoacoustic procedures are available to assess temporal resolution; however, the clinical use of these procedures is minimal at best. Results of the present study show that the GIN test holds promise as a clinically useful tool in the assessment of temporal resolution in the clinical arena. PMID- 16377997 TI - Rapid word-learning in normal-hearing and hearing-impaired children: effects of age, receptive vocabulary, and high-frequency amplification. AB - OBJECTIVE: This study examined rapid word-learning in 5- to 14-year-old children with normal and impaired hearing. The effects of age and receptive vocabulary were examined as well as those of high-frequency amplification. Novel words were low-pass filtered at 4 kHz (typical of current amplification devices) and at 9 kHz. It was hypothesized that (1) the children with normal hearing would learn more words than the children with hearing loss, (2) word-learning would increase with age and receptive vocabulary for both groups, and (3) both groups would benefit from a broader frequency bandwidth. DESIGN: Sixty children with normal hearing and 37 children with moderate sensorineural hearing losses participated in this study. Each child viewed a 4-minute animated slideshow containing 8 nonsense words created using the 24 English consonant phonemes (3 consonants per word). Each word was repeated 3 times. Half of the 8 words were low-pass filtered at 4 kHz and half were filtered at 9 kHz. After viewing the story twice, each child was asked to identify the words from among pictures in the slide show. Before testing, a measure of current receptive vocabulary was obtained using the Peabody Picture Vocabulary Test (PPVT-III). RESULTS: The PPVT-III scores of the hearing-impaired children were consistently poorer than those of the normal hearing children across the age range tested. A similar pattern of results was observed for word-learning in that the performance of the hearing-impaired children was significantly poorer than that of the normal-hearing children. Further analysis of the PPVT and word-learning scores suggested that although word-learning was reduced in the hearing-impaired children, their performance was consistent with their receptive vocabularies. Additionally, no correlation was found between overall performance and the age of identification, age of amplification, or years of amplification in the children with hearing loss. Results also revealed a small increase in performance for both groups in the extended bandwidth condition but the difference was not significant at the traditional p = 0.05 level. CONCLUSIONS: The ability to learn words rapidly appears to be poorer in children with hearing loss over a wide range of ages. These results coincide with the consistently poorer receptive vocabularies for these children. Neither the word-learning or receptive-vocabulary measures were related to the amplification histories of these children. Finally, providing an extended high-frequency bandwidth did not significantly improve rapid word learning for either group with these stimuli. PMID- 16377998 TI - Estimating noise-induced permanent threshold shift from audiometric shape: the ISO-1999 model. AB - OBJECTIVE: To describe the relation between audiometric shape and noise-induced permanent threshold shift averaged across the speech frequencies (N5123). DESIGN: Using an international standard (ISO-1999), 270 audiograms were created, representing both sexes and a variety of ages, exposure levels, and percentiles. Bulge depth (BD) was defined as the difference between pure-tone average for 2, 3, and 4 kHz and PTA for 1 and 6 kHz. RESULTS: N5123 was well-predicted by quadratic functions of BD, which accounted for 72 to 95% of the variance in N5123. CONCLUSIONS: Estimates of N5123, based on BD, can be helpful in medical legal diagnosis and allocation of hearing loss. PMID- 16377999 TI - How does the sound pressure generated by circumaural, supra-aural, and insert earphones differ for adult and infant ears? AB - OBJECTIVE: To determine how the ear canal sound pressure levels generated by circumaural, supra-aural, and insert earphones differ when coupled to the normal adult and infant ear. DESIGN: The ratio between the sound pressure generated in an adult ear and an infant ear was calculated for three types of earphones: a circumaural earphone (Natus Medical, ALGO with Flexicoupler), a supra-aural earphone (Telephonics, TDH-49 with MXAR cushion), and an insert earphone placed in the ear canal (Etymoup and down arrow tic Research, ER-3A). The calculations are based on (1) previously published measurements of ear canal impedances in adult and infant (ages 1, 3, 6, 12, and 24 months) ears (Keefe et al., 1993, Acoustic Society of America, 94:2617-2638), (2) measurements of the Thevenin equivalent for each earphone configuration, and (3) acoustic models of the ear canal and external ear. RESULTS: Sound-pressure levels depend on the ear canal location at which they are measured. For pressures at the earphone: (1) Circumaural and supra-aural earphones produce changes between infant and adult ears that are less than 3 dB at all frequencies, and (2) insert earphones produce infant pressures that are up to 15 dB greater than adult pressures. For pressures at the tympanic membrane: (1) Circumaural and supra-aural earphones produce infant pressures that are within 2 dB of adult ears at frequencies below 2000 Hz and that are 5 to 7 dB smaller in infant ears than adult ears above 2000 Hz, and (2) insert earphones produce pressures that are 5 to 8 dB larger in infant ears than adult ears across all audiometric frequencies. CONCLUSIONS: Sound pressures generated by all earphone types (circumaural, supra-aural, and insert) depend on the dimensions of the ear canal and on the impedance of the ear at the tympanic membrane (e.g., infant versus adult). Specific conclusions depend on the location along the ear canal at which the changes between adult and infant ears are referenced (i.e., the earphone output location or the tympanic membrane). With circumaural and supra-aural earphones, the relatively large volume of air within the cuff of the earphone dominates the acoustic load that these earphones must drive, and differences in sound pressure generated in infant and adult ears are generally smaller than those with the insert earphone in which the changes in ear canal dimensions and impedance at the tympanic membrane have a bigger effect on the load the earphone must drive. PMID- 16378000 TI - Evaluation of streamlined programming procedures for the Nucleus cochlear implant with the Contour electrode array. AB - OBJECTIVE: The objective of this study was to evaluate streamlined programming procedures for the Nucleus cochlear implant system with the Contour electrode array. DESIGN: Phase 1 involved an examination of the clinical MAPs for the first 103 recipients implanted with the Contour electrode array in the Melbourne Cochlear Implant Clinic, to examine the ability to predict the entire MAP based on a smaller number of clinically determined T- and/or C-levels. In phase 2, a subset of the streamlined procedures was selected and clinically evaluated, using speech perception and subjective preference measures. In the first study, the clinical MAP was compared with a MAP based on interpolating across three behavioral T-levels and three behavioral C-levels in a group of newly implanted subjects. The second study investigated the use of a single interpolated profile as the basis to creating the entire MAP. Initial evaluation compared the clinical MAP with two streamlined MAPs, one in which the C-level profile was derived from interpolation across a subset of T-levels and one in which the T-level profile was derived from interpolation across a subset of C-levels. In this case, the interpolated profile was based on five behavioral measures. Subsequently, the use of either three or a single T-level measure as the basis for the interpolated T level profile was evaluated. Eighteen subjects, who were experienced with the clinical MAP before enrollment in the study, participated in the initial evaluation. The subjects were selected to include a group whose RMS deviation from clinical MAP levels, as determined in Phase 1, was greater than that of the wider population. RESULTS: The Phase 1 analysis showed that as expected, larger differences were observed between the clinical and derived MAP levels as interpolation was applied across fewer measured electrodes and that the use of a single interpolated profile to create the entire MAP resulted in the greatest deviation. No significant group mean difference was found in speech perception scores for newly implanted subjects when mapped with the clinical versus the streamlined MAP based on three behavioral T- and three behavioral C-level measures. For some individual subjects, scores were higher with the streamlined MAP. Subjective reports from the comparative performance questionnaire were consistent with these findings. No significant group mean difference in speech perception scores was found in comparing the clinical MAP with the streamlined MAPs based on a single interpolated T- or C-level profile created from five behavioral measures. Individual effects were observed; however, there was no consistent finding across subjects. The use of three rather than five behavioral T-level measures in the procedure did not result in significantly lower group mean scores; however, significantly poorer scores were obtained for three of the 10 individual subjects. The use of a MAP based on a single behavioral measure did result in poorer speech perception scores when compared with the MAP based on five behavioral T-level measures. These findings were consistent with subjective results from the performance questionnaires administered to determine preference for program across a range of listening situations. CONCLUSIONS: Two streamlined programming procedures are recommended for use in the clinical setting: (1) interpolating across three measured T-levels and three measured C-levels and (2) interpolating across five measured T- or C-levels and using the interpolated profile for fitting of the alternative profile. PMID- 16378001 TI - Speech recognition and comfort using hearing instruments with adaptive directional characteristics in asymmetric listening conditions. AB - OBJECTIVE: Hearing instruments with adaptive directional microphone systems attempt to maximize speech-to-noise ratio (SNR) and thereby improve speech recognition in noisy backgrounds. When instruments with adaptive systems are fitted bilaterally, there is the potential for adverse effects as they operate independently and may give confusing cues or disturbing effects. The present study compared speech recognition performance in 16 listeners fitted bilaterally with the Phonak Claro hearing instrument using omni-directional, fixed directional, and adaptive directional microphone settings as well as mixed microphone settings (an omni-directional microphone on one side and an adaptive directional microphone on the other). DESIGN: Under anechoic conditions, speech was always presented from a loudspeaker directly in front of the listener (0 degree azimuth) whereas noise was presented from one or two loudspeakers arranged either symmetrically (0, 180, 90 + 270 degrees) or asymmetrically (170 + 240 degrees and 120 + 190 degrees) in the horizontal plane. Adaptive sentence recognition in noise measurement was supplemented by quality ratings. RESULTS: With symmetrical omni-directional settings (Omni/Omni), performance was poorer than a control group of 14 listeners with normal hearing tested unaided: Aided listeners required 4.3 dB more favorable SNR for criterion performance. In all loudspeaker arrangements in which directional characteristics could be exploited, performance with symmetrical adaptive microphones (Adapt/Adapt) was similar to the control group. The mixed microphone settings did not appear to confer any particular disadvantage for speech recognition from their asymmetric nature, always giving scores significantly better than Omni/Omni. Quality rating scores were consistent with speech recognition performance, showing benefits in terms of clarity and comfort for the Adapt/Adapt and Fixed/Fixed microphone conditions over the Omni/Omni and mixed microphone conditions wherever directional characteristics could be used. Similarly, the mixed microphone conditions were rated more comfortable and quieter for the noise than Omni/Omni. CONCLUSIONS: It is concluded that bilateral hearing instruments with adaptive directional microphones confer benefits in terms of speech recognition in noise and sound quality. Independence of the two adaptive control systems does not appear to cause untoward effects. PMID- 16378005 TI - Developments in the apheresis procedure for the treatment of inflammatory bowel disease. AB - Initially used to treat rheumatoid arthritis, nonselective therapeutic leukocytapheresis was applied to the treatment of inflammatory bowel disease (IBD) as early as the 1980s. Since then, the process has been further refined and 2 blood perfusion systems using membrane filtration are presently employed in Japan and Europe for the selective removal of leukocytes in patients with IBD: Cellsorba is a column of polyethylenephtarate fibers that captures lymphocytes and granulocytes, and Adacolumn is a column of cellulose acetate beads that selectively adsorb granulocytes and monocytes. These systems overcome the limitations of centrifugation. Leukocytapheresis has been shown to exert an overall anti-inflammatory effect, as peripheral leukocytes demonstrated a diminished capacity to produce inflammatory cytokines such as tumor necrosis factor-alpha, interleukin (IL)-1, IL-6, IL-8, and IL-1beta. In addition, down regulation in the expression of adhesion molecule L-selectin and a shift toward a more immature granulocyte phenotype were observed in the peripheral blood. The safety and beneficial therapeutic effect of leukocytapheresis in IBD are being investigated further. PMID- 16378006 TI - Preliminary data on the use of apheresis in inflammatory bowel disease. AB - In patients with inflammatory bowel disease (IBD), centrifugation has been attempted to remove leukocyte components from whole blood; however, the use of selective filters has proved to result in more active modification of cellular immunity in that 4 times as many white blood cells are removed, which may result in a greater therapeutic effect. Selective apheresis for treatment of IBD, in particular ulcerative colitis (UC), has been used in Japan and some European countries for several years; pilot studies with Adacolumn, a selective therapeutic granulocyte/monocyte apheresis device, in patients with IBD have recently been completed in the United States with favorable results. Unlike conventional pharmacological treatments, selective apheresis may be associated with a relatively low rate of adverse events. Multiple studies have suggested that selective apheresis may be of benefit as a steroid-sparing treatment. In an unblinded randomized trial in 69 steroid-dependent patients with active UC randomized to selective apheresis with Adacolumn or an increased dose of prednisolone, 83% of patients receiving Adacolumn achieved remission compared with 65% of those receiving an increased dose of prednisolone. In another uncontrolled study of 60 patients with active UC, treatment with Adacolumn selective apheresis enabled nearly 70% of steroid-dependent patients to discontinue prednisolone. An unblinded randomized controlled trial of a different selective apheresis device (Cellsorba) versus high-dose prednisolone in patients with active UC showed a greater therapeutic effect (74%) than high-dose prednisolone (38%) and lower frequency of adverse effects (24% versus 68%). PMID- 16378007 TI - Inflammatory bowel disease: epidemiology, pathogenesis, and therapeutic opportunities. AB - Ulcerative colitis (UC) and Crohn's disease (CD), the primary constituents of inflammatory bowel disease (IBD), are precipitated by a complex interaction of environmental, genetic, and immunoregulatory factors. Higher rates of IBD are seen in northern, industrialized countries, with greater prevalence among Caucasians and Ashkenazic Jews. Racial gaps are closing, indicating that environmental factors may play a role. IBD is multigenic, with the most clearly established genetic link between certain NOD2 variants and CD. Regardless of the underlying genetic predisposition, a growing body of data implicates a dysfunctional mucosal immune response to commensal bacteria in the pathogenesis of IBD, especially CD. Possible triggers include a chronic inflammatory response precipitated by infection with a particular pathogen or virus or a defective mucosal barrier. The characteristic inflammatory response begins with an infiltration of neutrophils and macrophages, which then release chemokines and cytokines. These in turn exacerbate the dysfunctional immune response and activate either TH1 or TH2 cells in the gut mucosa, respectively associated with CD and, less conclusively, with UC. Elucidation of immunological and genetic factors indicate multiple points at which the inflammatory cascade may be interrupted, yielding the possibility of precise, targeted therapies for IBD. PMID- 16378009 TI - Intraocular pressure measurement-comparison of dynamic contour tonometry and goldmann applanation tonometry. AB - PURPOSE: The dynamic contour tonometer (DCT, Pascal tonometer, Swiss Microtechnology AG, Port, Switzerland) was recently introduced as a new method of intraocular pressure measurement, supposedly independent of corneal properties. In this study we analyzed the agreement and correlation of dynamic contour tonometry and Goldmann applanation tonometry (GAT) and investigated the influence of central corneal thickness (CCT) and corneal curvature. We also considered preferential patient groups for both methods. METHODS: In a prospective study of 100 eyes without glaucoma, intraocular pressure was measured using dynamic contour tonometry and Goldmann applanation tonometry, followed by measurements of central corneal thickness and corneal curvature. RESULTS: A clear correlation between dynamic contour tonometry and Goldmann applanation tonometry was found (r = 0.693; P < 0.001). Dynamic contour tonometry generally resulted in higher intraocular pressure measurements (median difference + 1.8 mm Hg, mean difference + 2.34 mm Hg). Unlike dynamic contour tonometry, Goldmann applanation tonometry was remarkably affected by central corneal thickness, but neither method was significantly influenced by corneal curvature. Bland-Altman graphs showed remarkable disagreement between dynamic contour tonometry and Goldmann applanation tonometry, which could be partially explained by the influence of central corneal thickness on Goldmann applanation tonometry. To obtain valid readings, dynamic contour tonometry required a more extensive selection of patients than Goldmann applanation tonometry. CONCLUSIONS: Dynamic contour tonometry seems to be a reliable method for intraocular pressure measurement, which unlike Goldmann applanation tonometry is not influenced by central corneal thickness. In clinical practice, advantages from dynamic contour tonometry can be expected for cooperative patients, outpatients, and patients with sufficient bilateral ocular fixation, whereas Goldmann applanation tonometry measurements are more reliable in case of patients with inadequate cooperation, poor vision, or nystagmus. PMID- 16378010 TI - Outcomes of inferonasal Baerveldt glaucoma drainage implant surgery. AB - PURPOSE: To determine the outcomes of inferonasal Baerveldt glaucoma implant (BGI) surgery. PATIENTS AND METHODS: A retrospective, non-comparative case series of 182 eyes of 182 patients who underwent inferonasal placement of a Baerveldt glaucoma implant. The main outcome measures included intraocular pressure, number of glaucoma medications, best corrected visual acuity, and complications. RESULTS: Patients were followed for an average (+/- SD) of 19 +/- 15 months. Median Snellen visual acuity was unchanged at last follow-up. Intraocular pressure was reduced from a mean (+/- SD) of 28.6 +/- 11.5 mm Hg preoperatively to 13.4 +/- 5.7 mm Hg at most recent follow-up (P < 0.001). The number of antiglaucoma medications was reduced from a mean (+/- SD) of 2.7 +/- 1.3 preoperatively to 1.1 +/- 1.1 at most recent follow-up. Nineteen eyes met our criteria for failure, yielding a cumulative percent survival of 92% at 12 months, 88% at 24 months, and 77% at 44 months. The most common complications were hyphema (14 eyes, 8%), choroidal effusion (12 eyes, 7%), and corneal decompensation (19 eyes, 10%). Endophthalmitis and diplopia occurred rarely (1 eye, <1%; 3 eyes, 2%, respectively). CONCLUSIONS: Inferonasal Baerveldt glaucoma implant placement appears to be a safe and effective surgical option that may be helpful in certain clinical situations. PMID- 16378011 TI - Coding of glaucoma for patients with uveitis in the medicare database. AB - PURPOSE: To investigate possible causes for, and implications of, variations in coding of glaucoma for patients with uveitis in the Medicare database. PATIENTS AND METHODS: We identified 6,391 patients coded with one of several ICD-9 codes designating a form of uveitis from a 5% sample of the 1999 Medicare database. The proportion of patients with uveitis codes who also had codes for uveitic glaucoma, primary open-angle glaucoma (POAG), or unspecified open-angle glaucoma (OAG) were determined. The proportion with codes for argon laser trabeculoplasty (ALT), trabeculectomy, visual field examinations, fundus photographs, and gonioscopy were calculated in three groups: patients with both uveitis and uveitic glaucoma; those with both uveitis and POAG; and those with both uveitis and unspecified open-angle glaucoma. Summaries of the total allowed reimbursement related to eye visits and to eye codes were determined. RESULTS: Of the 6,391 patients with uveitis who were studied, 1,260 (19.7%) also had the code for POAG; 412 (6.4%) had the code for OAG; and 117 (1.8%) had the code for uveitic glaucoma. Visual field testing was less common among patients with uveitic glaucoma (37%) than among those with POAG/uveitis (46%) or OAG/uveitis (44%) (P < 0.001). There were no statistically significant differences among the three groups in the amount of total reimbursement related to eye services or eye codes. CONCLUSIONS: The code for POAG appears to be used in lieu of uveitic glaucoma in many cases. Discrepancies are probably not influenced by expected reimbursements, as much as by a lack of understanding about the available ICD-9 codes. PMID- 16378012 TI - Visual evoked potentials of the blue-sensitive pathway under cold provocation in normals and glaucomas. AB - BACKGROUND: Vascular dysregulation in open-angle glaucomas can be identified by measuring the blood circulation during exposure to cold water. Aim of this study was to find out whether the same cold stimulus would lead to functional changes in the visual evoked potentials (VEP) of the short-sensitive pathway in normals and glaucoma patients. PATIENTS AND METHODS: Blue-on-yellow pattern-visual evoked potentials were studied in 22 healthy control persons and 47 patients with primary open-angle glaucoma (25 high-pressure glaucomas, 22 normal pressure glaucomas with clinical signs of vasospastic hyperreactivity). A blue stripe pattern, presented in onset-offset mode on a yellow adaptation light served as the stimulus. Cold provocation was initiated by dipping one hand of the patient into cold water of 3 degrees C to 5 degrees C. The onset amplitudes and peak times were measured without cold exposition as well as two and four minutes after the cold exposition began. RESULTS: In the group of vasospastic glaucoma patients a significant reduction of visual evoked potential-amplitudes was observed during cold provocation (amplitude before ice exposition: 4.17 microV, amplitude following two minutes of ice exposition: 3.52 microV; paired test: P < 0.01). Other subject groups showed no significant amplitude reductions after cold provocation. Peak times of both open-angle glaucoma groups (132.3 +/- 18.7 milliseconds in high pressure, 132.7 +/- 14.5 milliseconds in normal pressure) were significantly increased in comparison to normals (117.4 +/- 8.0 milliseconds). However, no significant influence of the cold provocation on peak times could be found in all groups. CONCLUSION: Peak times of the blue-on-yellow visual evoked potentials are significantly prolonged in patients with primary open-angle glaucomas. Cold provocation causes a significant amplitude reduction of the blue-on-yellow visual evoked potential in the present normal-pressure glaucoma patients and reflects vascular dysregulation in patients with vasospastic hyperreactivity. PMID- 16378013 TI - Effect of cataract on frequency doubling perimetry in the screening mode. AB - PURPOSE: To determine the effect of cataract on frequency doubling perimetry in the screening mode. METHODS: Frequency doubling technology perimetry in the C-20 5 screening mode was performed prospectively on 34 patients with otherwise healthy eyes before and after phacoemulsification. Cataracts were scored using the LOCS III classification, and the number of missed points (at any level of severity) on the screening field was compared before and after surgery. RESULTS: Nine patients missed at least one point pre-operatively and had no misses post operatively. Those patients with posterior subcapsular (PSC) cataract were significantly more likely to miss points on the screening frequency doubling technology perimetry than those without posterior subcapsular cataracts (P < 0.001; Fisher exact test). In addition, the degree of posterior subcapsular cataract strongly correlated with the number of missed points pre-operatively (r = 0.97). The degree of nuclear sclerotic cataract did not correlate strongly with the number of missed points (r = 0.26), but the correlation was slightly stronger in the presence of cortical cataract (r = 0.36). CONCLUSION: Advanced age-related cataracts may cause false positive results on the screening frequency doubling technology, and the presence of even mild posterior subcapsular cataract may be a cause of false positive results and should be taken into consideration when interpreting the screening frequency doubling technology perimetry results. PMID- 16378014 TI - Comparison of postoperative refractive outcome in phacotrabeculectomy and phacoemulsification with posterior chamber intraocular lens implantation. AB - PURPOSE: To compare the postoperative refractive outcomes following phacotrabeculectomy and phacoemulsification with posterior chamber intraocular lens implantation. METHODS: A retrospective comparative study of 90 consecutive patients (95 eyes) with cataract with or without co-existing glaucoma who had undergone uncomplicated phacotrabeculectomy or phacoemulsification by a single surgeon. The biometry prediction error was determined for each case by the difference between the actual postoperative refraction and the preoperative predicted refractive outcome, in spherical equivalent. This was followed by a comparative analysis of the mean biometry prediction error, and the number of cases with error >0.50 D or 1.00 D. All biometry measurements were performed by one of the two qualified optometrists. RESULTS: The mean and absolute mean biometry prediction error of the 25 eyes that received phacotrabeculectomy (+0.20 D, absolute error 0.96 D) was comparable to the 70 eyes that received phacoemulsification (-0.14 D, absolute error 0.68 D), P = 0.18 (absolute error, P = 0.12). The proportion of cases with prediction error >0.50 D or 1.00 D was 60% or 40% for phacotrabeculectomy, and 44% or 17% for phacoemulsification, respectively. Phacotrabeculectomy was more likely to have a prediction error >1.00 D (P = 0.02) and a myopic shift of >0.50 D or 1.00 D (P = 0.03 or 0.02, respectively). No significant differences were found with regard to the frequency of hyperopic shift or prediction error >0.50 D. There was no significant difference in either the mean error or the frequency of the different types of error for the different biometry operators, types of glaucoma, or postoperative intraocular pressure control success status. CONCLUSION: Myopically shifted prediction error was significantly more frequent following posterior chamber intraocular lens implantation with phacotrabeculectomy compared with phacoemulsification, even when surgery was uncomplicated and performed by the same surgeon. PMID- 16378015 TI - Accuracy of an electronic monitoring and reminder device for use with travoprost eye drops. AB - PURPOSE: To evaluate the accuracy of a prototype electronic device for recording eye drop usage. PARTICIPANTS AND METHODS: Ten volunteers were randomly assigned to one of five usage patterns designed to mimic common patterns of use in glaucoma patients from 100% compliant to 50% compliant. All participants agreed to adhere to a pre-determined "dosing" schedule for 15 days using the monitoring/reminder device to instill artificial tears. Participants also recorded drop usage in a diary. The main outcome measures were device accuracy and reproducibility. Device accuracy was defined as the magnitude of the difference between the diary and device output for three variables: date, number of drops, and instillation time. RESULTS: Date stamping by the device was 100% accurate. The mean +/- SD time difference between the device and the diary was 2.0 +/- 19.7 minutes when data from all participants was pooled. In seven of the ten participants, the device did not record at least one drop. The mean +/- SD difference in the number of drops recorded by the device minus the diary was 0.16 +/- 0.97 when data from all participants was pooled. CONCLUSIONS: The prototype compliance reminder/monitoring device may underestimate compliance in some patients. The date and time stamping mechanisms were generally accurate and reproducible. PMID- 16378016 TI - Prediction of visual field defects on standard automated perimetry by screening C 20-1 frequency doubling technology perimetry. AB - PURPOSE: To determine if perimetric defects, initially seen with C-20-1 frequency doubling technology (FDT) in eyes with normal standard automated perimetry (SAP), will subsequently develop on standard automated perimetry. METHODS: Review of patients considered high-risk glaucoma suspects (GS) or with ocular hypertension (OHT), who underwent one C-20-1 FDT in 1997 or 1998. An abnormal frequency doubling technology was defined in two ways, as having either at least one, or at least two abnormal locations. An abnormal standard automated perimetry was defined as fulfilling two of three modified Anderson criteria for abnormality on two consecutive tests and at final standard automated perimetry. RESULTS: Of 63 eyes (50 patients), 18 (28.6%) had >or=1 FDT defects, and 12 (19.0%) had >or=2 defects. After follow-up of 62 +/- 26 months, an abnormal standard automated perimetry developed in 5 eyes (28%) with frequency doubling technology defects at 22 +/- 13 months (28% of eyes with >or=1 defect and 42% of eyes with >or=2 FDT defects), and in 4 of 45 eyes (9%) with normal frequency doubling technology at 32 +/- 30 months. Kaplan-Meier survival analysis revealed the risk of an abnormal standard automated perimetry at 5 years with or=1 defects, and >or=2 defects, was 10.3%, 11.8%, 30.6%, and 46.4% respectively (P = 0.060 for >or=1 defects compared with no defects; P = 0.002 for >or=2 defects compared with or=20% IOP reduction and criteria B, IOP < 15 mm Hg with >or=30% IOP reduction. Kaplan-Meier survival analysis and Cox's proportional hazards regression analysis were used to estimate and determine influence of laser suture lysis on success rates after trabeculectomy. RESULTS: Laser suture lysis was performed in 95 eyes (84 patients) whereas 146 eyes (121 patients) did not require it. IOP was significantly higher at 12 months in laser suture lysis eyes compared with non-laser suture lysis eyes (12.9 +/- 5.2 mm Hg versus 11.0 +/- 4.1 mm Hg; P = 0.04). Laser suture lysis eyes also had decreased success rates with criteria B (57% compared with 73% at 1 year for non-LSL eyes; P = 0.005). However, only late laser suture lysis (last LSL session >10 days after trabeculectomy) was associated with worse outcome in Cox's proportional hazards model using criteria B (HR = 2.26; P = 0.004). Late hypotony occurred in 4% and 1% of non-LSL and LSL eyes, respectively. CONCLUSIONS: Eyes that undergo laser suture lysis >10 days after initial trabeculectomy with mitomycin C are more likely to have poorer long-term IOP control than eyes not requiring laser suture lysis or eyes undergoing laser suture lysis .1) and from 71.1% to 79.0% (p > .25). Sensitivities of other screening tests were unaffected by changing the criterion standard. CONCLUSIONS: Criterion standards that miss the same CIN 2 or worse as the screening tests likely cause inflation of the sensitivity of those tests. PMID- 16378027 TI - Digitized cervical images: problems, solutions, and potential medical impact. AB - OBJECTIVE: To demonstrate compression, illumination enhancement, registration, segmentation, automated classification and steganography using digitized cervical images. MATERIALS AND METHODS: The Hybrid Multi-Scale Vector Quantization algorithm developed at Texas Technological University and other automated systems were used to improve digitized cervical images. RESULTS: We demonstrated high levels of image compression, illumination enhancement, registration, automated segmentation and classification and steganography of digitized cervical images. CONCLUSIONS: Digitized cervical images can be altered to facilitate research of cervical neoplasia. PMID- 16378028 TI - Digital tools for collecting data from cervigrams for research and training in colposcopy. AB - Colposcopy is a critical part of gynecologic practice but has documented deficiencies, including lack of correlation between the colposcopic appearance and the severity of underlying neoplasia, limited reproducibility, and difficulty in the optimal placement of colposcopically directed biopsies. In a collaborative effort to improve colposcopy, we are analyzing digitized cervigram images from National Cancer Institute-funded studies. Specifically, the National Cancer Institute has collected close to 100,000 cervigrams, digitized to create a database of images of the uterine cervix for research, training, and education. In addition to the cervigram images, this database contains clinical, cytologic, and molecular information at multiple examinations of 15,000 women, with password and ID labeling strategies to protect patient privacy. The National Library of Medicine has designed two web-accessible software tools. The Boundary Marking Tool allows experts on colposcopy to perform an evaluation of the pictures and to mark boundary regions of normal and abnormal regions of the uterine cervix; these evaluations are collected and saved in the database. The Multimedia Database Tool enables retrieval of test and image biomedical data according to specific queries, for example, all women with cervical intraepithelial neoplasia 3 whose cytologic results are atypical squamous cells of undetermined significance. The resource soon will be available as an open resource, via a teaching tool coordinated by a database manager, which will permit a variety of applications for teaching and research. In this article, we describe the perceived need for the resource and its components. PMID- 16378029 TI - Refining the management of low-grade cervical abnormalities in the UK National Health Service and defining the potential for human papillomavirus testing: a commentary on emerging evidence. PMID- 16378030 TI - Preparing digitized cervigrams for colposcopy research and education: determination of optimal resolution and compression parameters. AB - OBJECTIVE: Visual assessment of digitized cervigrams through the Internet needs to be optimized. The National Cancer Institute and National Library of Medicine are involved in a large effort to improve colposcopic assessment and, in preparation, are conducting methodologic research. MATERIALS AND METHODS: We selected 50 cervigrams with diagnoses ranging from normal to cervical intraepithelial neoplasia 3 or invasive cancer. Those pictures were scanned at 5 resolution levels from 1,550 to 4,000 dots per inch (dpi) and were presented to 4 expert colposcopists to assess image quality. After the ideal resolution level was determined, pictures were compressed at 7 compression ratios from 20:1 to 80:1 to determine the optimal level of compression that permitted full assessment of key visual details. RESULTS: There were no statistically significant differences between the 3,000 and 4,000 dpi pictures. At 2,000 dpi resolution, only one colposcopist found a slightly statistically significant difference (p = 0.02) compared with the gold standard. There was a clear loss of quality of the pictures at 1,660 dpi. At compression ratio 60:1, 3 of 4 evaluators found statistically significant differences when comparing against the gold standard. CONCLUSIONS: Our results suggest that 2,000 dpi is the optimal level for digitizing cervigrams, and the optimal compression ratio is 50:1 using a novel wavelet-based technology. At these parameters, pictures have no significant differences with the gold standard. PMID- 16378031 TI - Distribution of cervical intraepithelial neoplasia 2, 3 and cancer on the uterine cervix. AB - OBJECTIVE: To determine the distribution of cervical intraepithelial neoplasia 2 or 3 on the uterine cervix. MATERIALS AND METHODS: During two screening studies, 5,060 women underwent colposcopy with biopsy. If a cervical quadrant had no lesion, a random biopsy at the squamocolumnar junction was obtained. RESULTS: Cervical intraepithelial neoplasia 2 or worse was more common on the posterior (426/806; 52.9%) than on the anterior (380/806; 47.2%) cervix (p < .05) and was equally likely on the right (411/806; 51.0%) and left (395/806; 49.0%) sides of the cervix. More quadrants with colposcopically detected lesions of human papillomavirus or worse came from the anterior (1,338; 57.5%) than posterior (991; 42.6%) cervix (p < .05), whereas similar numbers of these lesions were found on the left (1,163) and right (1,166) sides of the cervix. CONCLUSIONS: Cervical intraepithelial neoplasia 2 or worse is slightly more common on the posterior cervix. Using colposcopically detected lesions as a surrogate for cervical intraepithelial neoplasia 2 or worse inflates the prevalence of cervical intraepithelial neoplasia on the anterior cervix. PMID- 16378032 TI - Internal Federation for Cervical Pathology and Colposcopy nomenclature and treatment for cervical preinvasive disease. AB - OBJECTIVE: To examine the effects of IFCPC transformation zone type and treatment modality on outcome from excisional treatments for cervical pre-invasive disease at an inner city colposcopy department. MATERIALS AND METHODS: All patients treated within the colposcopy department for whom age, treatment modality, resection margin status at histology and six-month cytologic data was available. RESULTS: Transformation zone type is affected by patient age. Change in accessibility of the transformation zone to examiantion by colposcopy affected cytologic outcome. There was no obvious effect of treatment modality to explain this change in outcome. CONCLUSIONS: Transformation zone type is an important determinant of excisional treatment outcome for cervical disease. This needs to be considered when comparing therapeutic success. PMID- 16378033 TI - Sebaceous hyperplasia of the vulva: case report and review of the literature. AB - Sebaceous hyperplasia is a cutaneous lesion consisting of soft, yellow, papular lesions usually occurring on the face in elderly patients. The occurrence of the lesion on the vulva is exceptionally rare. We were able to identify only three cases reported in the literature. We report a case of sebaceous hyperplasia of the vulva with an unusual presentation. PMID- 16378034 TI - Initiating the American Society for Colposcopy and Cervical Pathology Humanitarian Program: Peru 2005. PMID- 16378036 TI - Clinical question: ask the experts. A 45-year-old woman seeks treatment at your clinic with atypical squamous cells from Pap smear results. PMID- 16378038 TI - The American Society for Colposcopy and Cervical Pathology: Hagerstown, Maryland. PMID- 16378037 TI - Home Study Course: Winter 2006. AB - OBJECTIVE: The Home Study Course is intended for the practicing colposcopist or practitioner who is seeking to develop or enhance his or her colposcopic skills. The goal of the course is to present colposcopic cases that are unusual or instructive in terms of appearance, presentation, or management or that demonstrate new and important knowledge in the area of colposcopy or pathology. Participants may benefit from reading and studying the material or from testing their knowledge by answering the questions. ACCME ACCREDITATION: The American Society for Colposcopy and Cervical Pathology (ASCCP) is accredited by the Accreditation Council for Continuing Medical Education (ACCME) to provide continuing medical education for physicians. The ASCCP designates this continuing medical education activity for 1 hour Category I credit of the ASCCP's Program for Continuing Professional Development and the Physician's Recognition Award of the American Medical Association. Credit is available for those who choose to apply. The Home Study Course is planned and produced in accordance with the ACCME's Essential Areas and Elements. DISCLOSURE: Faculty must disclose any significant financial interest or relationship with proprietary entities that may have a direct relationship to the subject matter. For this course, the author had no such relationship to report. PMID- 16378039 TI - Proceedings of the 2005 International Congress on Respiratory Viruses. PMID- 16378040 TI - Air quality, tobacco smoke, urban crowding and day care: modern menaces and their effects on health. AB - BACKGROUND: The known adverse health effects of outdoor air pollution were reduced in the last century by effective legislation and pollution control. Although combustion of biomass fuels in the indoor environment remains a major hazard in developing countries, there has been a change in the nature of the traffic-generated air pollutants in outdoor air in developed countries. The role of day care and siblings in increasing the risk of infection early in life contrasts with protection from allergic disease later. METHODS: The mechanisms of how these pollutants exert their effects are poorly understood, but there is emerging evidence that the toxic effects may be related to infection. This synopsis discusses the epidemiologic relationship of the menaces of modern living, including air pollution, urban crowding and infection, and will explore some of the mechanisms of how they act synergistically to cause exacerbations of illnesses in individuals with preexisting respiratory conditions, such as asthma. It will also discuss the roles of day care and siblings in relation to respiratory disease risk. RESULTS: Current evidence suggests that much of the morbidity and mortality related to sources of both indoor and outdoor pollution occur by causing or interacting with respiratory infection and other acute or chronic health conditions, such as asthma. CONCLUSION: Improvements in air quality through efforts to tackle environmental problems such as pollution and tobacco smoke will help achieve better health. Day care and siblings increase infectious disease early in life but are associated with protection against development of allergic disease later. PMID- 16378041 TI - Virus-induced airway dysfunction: pathogenesis and biomechanisms. AB - BACKGROUND: Viral respiratory tract infections cause significant morbidity and mortality. Respiratory viruses are suspected to play a role in the inception of asthma early in life. Respiratory syncytial virus (RSV) is the most common cause of infant bronchiolitis, which is associated with the development of childhood wheezing and asthma. However, it is not clear whether this association is "causal" or "circumstantial." METHODS: Animal models have been pivotal in studying the pathophysiology of viral respiratory infections. Various approaches to assessing airway inflammation and function have been used to define the mechanisms of virus-induced airway dysfunction and to address clinically relevant questions regarding the role of RSV in wheezing and asthma after bronchiolitis. RESULTS: Viral lower respiratory tract infections alter airway function in humans and animals. The extent and duration of the alterations may depend on the virus itself, host factors and environmental factors. Animal studies demonstrated that viral infection induces airway hyperresponsiveness and enhances this alteration in the allergen-sensitized and exposed host. This altered airway function is mediated by immune and neurogenic inflammatory mechanisms. Recent studies in mice show that neonatal RSV infection sensitizes the newborn to develop an asthma-like phenotype on reinfection, providing further opportunities to investigate the role of RSV in postbronchiolitis wheezing and asthma in this animal model. CONCLUSIONS: Further studies are needed to fully establish the mechanisms underlying the pathophysiology of viral respiratory tract infections and to clarify their role in the inception and/or progression of chronic airway diseases such as asthma. The results of ongoing therapeutic studies promise to minimize the impact of such viral infections. PMID- 16378042 TI - Viral infections, cytokine dysregulation and the origins of childhood asthma and allergic diseases. AB - BACKGROUND: The origins of asthma and allergic disease begin in early life for many individuals. It is vital to understand the factors and/or events leading to their development. METHODS: The Childhood Origins of Asthma project evaluated children at high risk for asthma to study the relationships among viral infections, environmental factors, immune dysregulation, genetic factors, and the development of atopic diseases. Consequently wheezing illnesses, viral respiratory pathogen identification, and in vitro cytokine response profiles were comprehensively evaluated from birth to 3 years of age, and associations of the observed phenotypes with genetic polymorphisms were investigated. RESULTS: For the entire cohort, cytokine responses did not develop according to a strict T helper cell 1 or T helper cell 2 polarization pattern during infancy. Increased cord blood mononuclear cell phytohemagglutin-induced interferon-gamma responses of mononuclear cells were associated with decreased numbers of moderate to severe viral infections during infancy, especially among subjects with the greatest exposure to other children. In support of the hygiene hypothesis, an increased frequency of viral infections in infancy resulted in increased mitogen-induced interferon-gamma responses at 1 year of age. First year wheezing illnesses caused by respiratory viral infection were the strongest predictor of subsequent third year wheezing. Also, genotypic variation interacting with environmental factors, including day care, was associated with clinical and immunologic phenotypes that may precede the development of asthma. CONCLUSIONS: Associations between clinical wheezing, viral identification, specific cytokine responses and genetic variation provide insight into the immunopathogenesis of childhood asthma and allergic diseases. PMID- 16378043 TI - Factors predicting childhood respiratory syncytial virus severity: what they indicate about pathogenesis. AB - BACKGROUND: A plethora of clinical experience exists defining the factors associated with differences in severity of childhood respiratory syncytial virus (RSV) infections. These clinical severity factors reveal a wealth of information about the pathogenesis of this disease. Reviewing and interpreting the clinical risk factors to gain an insight into RSV pathogenesis is important, especially considering the relative lack of parallel between many animal models of RSV infection and observed human disease. METHODS: Existing and unpublished data on severity risk factors are reviewed and placed into a working pathogenesis model. RESULTS: Important factors discussed are: (1) the critical role of the timing of the infection; (2) its rapidity of progression to involve the lower respiratory tract; (3) factors limiting this spread; (4) the nature of the pathogenic immune response; and (5) the host genetic and other factors that alter this immune response. CONCLUSIONS: Pending new data involving these and other processes will more fully illuminate the spectrum of childhood RSV disease. PMID- 16378044 TI - Chronic manifestations of respiratory syncytial virus infection in premature infants. AB - BACKGROUND: Respiratory syncytial virus (RSV) infection in healthy infants born at term results in long term sequelae. Infants born prematurely are at increased risk of severe acute RSV infection; thus it would seem likely that such infants would be at increased risk of long term respiratory sequelae. METHODS: Methods of assessing the long term outcome of RSV infection are discussed and the results of retrospective and prospective studies investigating chronic respiratory morbidity after RSV infection in premature infants are reviewed. RESULTS: Documentation of all health care utilization, parental documentation of symptom status and lung function measurement provide a comprehensive and quantitative assessment of respiratory outcome. Studies that have included such outcome measures have demonstrated that RSV hospitalization in infants born between 32 and 35 weeks of gestational age and in those born more prematurely who developed chronic lung disease was associated with more hospital admissions, inpatient days, physician contacts and outpatient visits in the first 2 years after birth. Children born before 32 weeks of gestation who developed chronic lung disease also required more outpatient attendances and prescriptions and respiratory medications in years 2 through 4. Prospective data collection has demonstrated that chronic respiratory morbidity occurs in very premature infants, regardless of whether their RSV infection required hospitalization. CONCLUSION: Chronic respiratory morbidity is increased in premature infants after RSV infection. The duration of this increased morbidity and the impact of other viral infections, particularly dual infection with RSV, on long term sequelae merit investigation. PMID- 16378045 TI - Respiratory syncytial virus infections: old challenges and new opportunities. AB - BACKGROUND: Respiratory syncytial virus (RSV) is the most common cause of bronchiolitis and pneumonia among children aged < 1 year. The majority of children hospitalized for RSV infection are younger than 6 months of age. RSV also causes repeated infections including severe lower respiratory tract disease, which may occur at any age, especially among the elderly or those with compromised cardiac, pulmonary, or immune systems. METHODS: Using the mouse model of RSV infection, this article examines the immunopathogenesis during acute and chronic phases of the disease. This model allows for measurement of basal enhanced pause, which reflects airway obstruction in the acute phase, and the response to methacholine challenge to assess airway hyperresponsiveness during the chronic phase. This article also summarizes some recent studies focusing on novel perspectives and strategies for treatment and prevention of RSV infections. RESULTS: Compared with the lungs of sham-inoculated control mice, mice inoculated with live RSV showed a persistent progression of the severity of pneumonia as determined by an increasing histopathologic score. Mucus production of RSV infected mice in the acute phase illustrated increased periodic acid-Schiff positive hypertrophic cells in central and peripheral airways. CONCLUSIONS: RSV infected mice with persistent airway hyperresponsiveness exhibited the presence of abnormal chronic inflammatory changes and mucus overproduction, which likely contributed to long term airway disease induced by RSV infection. These findings provide a histologic correlation to the abnormal pulmonary responses documented by plethysmography. Current trials have demonstrated positive results in continuing to target different alternatives for a new RSV vaccine. PMID- 16378046 TI - State of the evidence for standard-of-care treatments for croup: are we where we need to be? AB - BACKGROUND: Croup is a term that groups several different clinical syndromes with inspiratory stridor. The failure to delineate the specific syndromes has led to suboptimal treatment in many instances. METHODS: A literature review and personal experience have been analyzed. RESULTS: Specific croup syndromes have been identified. Most croup steroid-treatment studies have failed to adequately identify the specific illness being treated. With 2 exceptions, all studies done to date have been too small to sufficiently evaluate risks of steroids if the risk is 1% or less. CONCLUSIONS: Three conclusions were reached: (1) no steroid treatment studies of moderate or severe laryngotracheitis have been of adequate size to determine the risk of progressive viral infection or secondary bacterial infection; (2) single dose steroid treatment or limited nebulized use is probably safe; and (3) we should encourage better clinical diagnosis of croup illnesses. PMID- 16378047 TI - Human metapneumovirus: a ubiquitous and long-standing respiratory pathogen. AB - BACKGROUND: Human metapneumovirus (hMPV) is a recently described human pathogen first identified in respiratory specimens of young children suffering from respiratory syndromes ranging from mild to severe. METHODS AND RESULTS: Virological studies have reported the presence of hMPV infections in many countries from all continents. Seroprevalence studies have indicated that the virus has been circulating in humans for more than 50 years and that it infects virtually all children by the ages of 5-10 years. In young children, hMPV has been mainly associated with bronchiolitis but also with pneumonitis, otitis media and acute exacerbation of asthma. The contribution of hMPV in respiratory syndromes of adults has been studied considerably less; initial studies have indicated a role for this pathogen in flu-like syndromes and in significant percentages of chronic obstructive pulmonary disease exacerbations and cases of community-acquired pneumonia during the winter-spring period. Both primate and rodent experimental models have been used to characterize the pathogenesis of this respiratory virus. In some of these models, intranasal hMPV inoculation has elicited not only important viral replication but also significant pulmonary inflammation and clinical disease. Recently a few groups have developed reverse genetic systems for hMPV, allowing a better understanding of viral pathogenesis and generation of attenuated viral strains for immunization. CONCLUSIONS: Recent studies on hMPV have provided a better understanding of the epidemiology and pathogenesis associated with this viral infection, and have enhanced the prospect of developing efficient therapeutic agents and vaccine candidates. PMID- 16378048 TI - Avian influenza: an omnipresent pandemic threat. AB - BACKGROUND: Humans have faced 3 major influenza pandemics in the 20th century. In recent years, it has become evident that domestic poultry play an important role in the generation of novel influenza strains with the capacity to cross the species barrier and infect and kill humans at an alarming rate. There is particular concern that avian influenza viruses of the H5N1 subtype could cause a pandemic. METHODS: A better understanding of the genetic factors that lead to interspecies transmission is essential to prevent the emergence of influenza pandemics. In addition, the stockpiling of antiviral drugs and development of vaccines against potentially pandemic viruses must be considered under the umbrella of pandemic plans. RESULTS: The world is ill-prepared to face an influenza pandemic. Only a handful of countries have developed influenza pandemic plans, and even fewer are developing vaccines or stockpiling antiinfluenza drugs to ameliorate the impact of a potential pandemic. Currently the major undertaking in several at risk nations is to implement effective control measures to stop the spread of the virus at its source, that is, avian species. These measures include the culling of domestic poultry to contain the virus, a practice that could eventually bring these countries to a financial and social breaking point. CONCLUSIONS: Avian influenza disease is preventable in humans and birds with the concerted effort of governments and poultry producers, large and small, to improve biosecurity and education programs. Pandemic plans can reduce the impact of the pandemic; however, preventing avian influenza in poultry can avert a pandemic altogether. PMID- 16378049 TI - Role of viral infections, atopy and antiviral immunity in the etiology of wheezing exacerbations among children and young adults. AB - BACKGROUND: At least 101 serotypes of rhinoviruses have been identified. Rhinoviruses have often been associated with trivial upper respiratory infections but more recent evidence suggests that they play a significant role in the pathogenesis of asthma exacerbations. METHODS: The impact of rhinoviruses as a cause of respiratory morbidity in infancy and the role of rhinoviruses in causing asthma exacerbations in the children and adult population is discussed. Past and current experimental and epidemiologic evidence was assessed to suggest a causal role rather than just an association. Mechanism of susceptibility to virus infection in asthmatics were discussed, as were data indicating important interactions between allergic sensitization and rhinovirus infections in risk of asthma exacerbations. RESULTS: Asthmatic individuals have increased severity and duration of lower airway symptoms and greater declines in lung function upon infection with rhinovirus, suggesting greater susceptibility to rhinovirus infection because of impaired antiviral immunity. Early virus-induced apoptosis, which aborts virus replication, was impaired in asthmatics. Treating allergic airway inflammation may reduce the frequency and severity of symptoms triggered by rhinoviruses. CONCLUSIONS: The evidence in children and young adults suggests that atopy and allergic inflammation in the airways may be critical determinants of an adverse response to rhinovirus. Increased awareness of the spectrum of age groups that are affected by rhinoviruses is needed to encourage the development of new treatments and strategies that may be helpful in reducing the frequency and severity of symptoms triggered by rhinoviruses. PMID- 16378050 TI - History and recent advances in coronavirus discovery. AB - Human coronaviruses, first characterized in the 1960s, are responsible for a substantial proportion of upper respiratory tract infections in children. Since 2003, at least 5 new human coronaviruses have been identified, including the severe acute respiratory syndrome coronavirus, which caused significant morbidity and mortality. NL63, representing a group of newly identified group I coronaviruses that includes NL and the New Haven coronavirus, has been identified worldwide. These viruses are associated with both upper and lower respiratory tract disease and are likely common human pathogens. The global distribution of a newly identified group II coronavirus, HKU1, has not yet been established. Coronavirology has advanced significantly in the past few years. The SARS epidemic put the animal coronaviruses in the spotlight. The background and history relative to this important and expanding research area are reviewed here. PMID- 16378051 TI - Preparing for the next influenza pandemic: lessons from multinational data. AB - BACKGROUND: In the past decade, avian influenza has made several incursions of increasing scope and virulence into humans. The likelihood of another pandemic is increasing with time. In work recently published, influenza was found to be the principal cause of the increase in mortality in the United States during the winter months. In a companion report, the U.S. national vaccination program was shown to have increased coverage of high risk groups 5-fold from 1980 to 1999, but excess mortality did not decline in any elderly age group. The Multinational Influenza Seasonal Mortality Study has assembled and has begun to mine mortality data from many countries. Early results indicate that the U.S. results extend to other economically developed countries and probably worldwide. RESULTS: The Multinational Influenza Seasonal Mortality Study data extend the observations of others that there were heralding events that provided advance warning for all of the pandemics of the 20th century. Moreover, in the first year of emergence of A(H3N2) viruses, the 1968-1969 pandemic produced little excess mortality outside of North America. It appears that there were at least 2 variants of the pandemic virus, differing at 1 or more internal gene loci, and that the more virulent form emerged as dominant in the second pandemic season. CONCLUSIONS: Integrating these findings, it seems clear that the influenza control strategy now used in about 50 countries is less than optimal. While it is likely that there will be more time to react in the pandemic season than previously imagined, an enhancement of the historical strategy is clearly indicated. Furthermore, the vaccine shortage that is presently inevitable suggests that a departure from the historical strategy if calamitous ineffectiveness is to be avoided. PMID- 16378052 TI - The role of macrophages in allograft rejection. AB - Macrophage accumulation has long been recognized as a feature of allograft rejection, yet the role of macrophages in rejection remains underappreciated. Macrophages contribute to both the innate and acquired arms of the alloimmune response and thus may be involved in all aspects of acute and chronic allograft rejection. Recent advances in macrophage biology have allowed a better understanding of the mechanisms of macrophage accumulation, their state of activation and the pleuripotent roles they play in allograft rejection. Therapeutic attention to macrophages, in addition to T lymphocytes, may lead to improved outcomes in organ transplantation. PMID- 16378053 TI - Fuzzy organ allocation system for cadaveric kidney transplantation. AB - BACKGROUND: The recipient selection decision for a cadaveric donor kidney is complex and based on multiple criteria, not only medical but also ethical and political criteria. METHODS: In this study, we develop the Fuzzy Organ Allocation System (FORAS) to determine who among potential recipients receives a cadaveric kidney when it becomes available. FORAS balances various kidney allocation objectives and deals with the ambiguity and fuzziness in the allocation process. RESULTS: We used simulation to investigate how well FORAS represents the thinking of a transplant physician with regard to kidney allocation. We also compared FORAS with the United Network for Organ Sharing (UNOS) scoring system and the Turkish National Coordination for Organ Transplant (TONKS) algorithm used in Turkey. We found that FORAS well represents expert thinking in kidney allocation. CONCLUSIONS: A simulated kidney allocation experiment based on real patient and donor data showed that FORAS is more useful than other kidney allocation systems because its results more closely reflect the thinking of experienced transplant physicians. PMID- 16378054 TI - Kidney allocation in the United States: fuzzy on the details? PMID- 16378055 TI - Up-to-date epidemiological data and better treatment for Kaposi's sarcoma. PMID- 16378056 TI - Islet transplantation: steeple chase and the next hurdle. PMID- 16378057 TI - Endothelial functions improve with decrease in asymmetric dimethylarginine (ADMA) levels after renal transplantation. AB - BACKGROUND: Chronic kidney disease (CKD) is associated with increased cardiovascular events. The relationships between the markers of inflammation and endothelial dysfunction were investigated both before and after living donor kidney transplantation. METHODS: Twenty-seven renal transplant patients were studied. Eleven patients (six male, five female) were on cyclosporine A, whereas 16 patients (nine male, seven female) were treated with tacrolimus based regimes. Twenty-seven subjects (12 males, 15 females) were studied as controls. Plasma adiponectin, high sensitive C reactive protein (hsCRP), Asymetric dimethyl arginine (ADMA) levels were studied before transplantation and on days 1, 3, 7, 14, and 28. The brachial artery flow mediated dilatation (FMD) was studied before transplantation and on the 28th day. RESULTS: Serum hsCRP and ADMA levels decreased significantly from the first posttransplantation day on each measurement (P<0.001 for all) while the decrement of plasma adiponectin started in the third day (P<0.001 for all). The FMD was lower in the patients than the control group (P<0.001) and improved significantly in the 28th day of transplantation (P<0.001). CONCLUSIONS: The results indicate that ADMA is associated with FMD in CKD both before and after kidney transplantation. Endothelial functions improve at the very beginning of the posttransplantation period with accompanying reduction in ADMA and hsCRP levels. PMID- 16378058 TI - Early and frequent histological recurrence of Crohn's disease in small intestinal allografts. AB - BACKGROUND: Recurrence of Crohn's disease in small intestinal allografts, although rarely described, can cause serious morbidity and jeopardize graft survival among transplant recipients with Crohn's disease. However, systematic studies to determine the frequency, predictors, and clinical implications of recurrent Crohn's disease have not been reported METHODS: We analyzed our transplant program's experience with small intestinal allografts in patients with Crohn's disease based on retrospective review of clinical and pathological records and corresponding pathology slides. RESULTS: Of 67 patients undergoing 70 transplantations between 1998 and 2004, six adults (three males, three females; mean age 48.1 years) had Crohn's disease complicated by short gut syndrome and total parenteral nutrition failure. Four survivors surveyed endoscopically for a mean 29 (range, 20-40) months and underwent a mean 37 endoscopic examinations with biopsies (range, 31-44) while on maintenance immunosuppression. Despite absence of any endoscopic or clinical manifestations of Crohn's disease throughout this period, two patients had granulomatous enteritis characteristic of Crohn's disease in multiple biopsies, one patient in 8/44 examinations (18%) ranging from 34 days to 20 months postoperatively and the other in 6/32 examinations (19%) ranging from 20 days to 22 months postoperatively. No comparable changes occurred in 57 other patients without Crohn's disease followed endoscopically under the same protocol CONCLUSIONS: Histological recurrence of Crohn's disease may occur in small intestinal allografts despite the absence of endoscopic and clinical disease manifestations. Such recurrences are probably not rare, may occur as early as 3 weeks after transplantation, and do not necessarily portend early clinical recurrence or mandate aggressive therapy to prevent allograft loss. PMID- 16378059 TI - Living-donor liver transplantation with renoportal anastomosis for patients with large spontaneous splenorenal shunts. AB - BACKGROUND: End-stage liver disease is often accompanied by large spontaneous splenorenal shunts and thrombosed portal vein. Renoportal anastomosis for spontaneous splenorenal shunts in living-donor liver transplantations is one of the solutions for the treatment of these patients. However, the long-term outcome, portal venous hemodynamics after liver transplantation, and the effects of altering the renal venous drainage remained unknown. METHODS: We performed three living-donor liver transplantations with renoportal anastomosis for the treatment of spontaneous splenorenal shunts between 1999 and 2004. We then evaluated the outcome of this procedure using short- and long-term follow-ups in which the postoperative graft function, renal function, radiological images and portal hemodynamics were examined. RESULTS: All three patients who underwent a living-donor liver transplantation with renoportal anastomosis are alive with normal graft function and a patent renoportal anastomosis. The portal hemodynamics were similar to those in conventional living-donor liver transplantation recipients, and had no harmful effect on allograft function. Left renal function returned to normal after the temporal impairment in two cases, and remained slightly impaired in one, although it was negligible clinically. CONCLUSIONS: Living-donor liver transplantation with renoportal anastomosis for the treatment of spontaneous splenorenal shunts in patients with end-stage liver disease is a life-saving and safe technique and should be discussed as a treatment option for patients with splenorenal shunts. PMID- 16378060 TI - Composite tissue allotransplantation: classification of clinical acute skin rejection. AB - BACKGROUND: Composite tissue allotransplantation (CTA) is a recently introduced option for limb replacement and reconstruction of other nonreconstructible tissue defects. As with recipients of other allotransplants, CTA recipients can experience rejection episodes that are presumed to be mediated by immune mechanisms similar to those affecting solid organ grafts. However, a systematic examination of this process has not been performed, and there are no standardized criteria for the description of severity or type of rejection METHODS: We collected biopsies from human limb allografts and abdominal walls in various stages of rejection for histological and immunohistochemical analysis to formulate a CTA rejection scheme. Biopsies were ranked by severity and reproducibility of the system was tested using a second set of biopsies. Tissue slides were examined blindly by three pathologists and the nonparametric Kendall coefficient of concordance (W) was used to assess the amount of agreement among the pathologists in their classification grades. RESULTS: Rejection initially appeared as a perivascular infiltrate progressing to involve the dermis. Arteritis was observed only in the medium to large size arteries of the subcutis. Myositis was seen occasionally. Perineural involvement without frank neuritis was present in advanced rejection. The infiltrate was predominantly CD4+ in milder cases and CD8+ in advanced cases. HLA-DR was minimally expressed in keratinocytes even in severe rejection. Kendall's W was 0.9375 (p4 and >7 months in the absence of exogenous immunosuppression; in two asplenic pigs, kidney grafts were rejected on days 4 and 15. CONCLUSIONS: Successful SpTx can result in hematopoietic cell engraftment and in vitro donor-specific unresponsiveness, enabling prolonged survival of subsequent donor-matched kidney grafts without immunosuppression. PMID- 16378078 TI - Serum sCD30 in monitoring of alloresponse in well HLA-matched cadaveric kidney transplantations. AB - In kidney transplantation, pretransplant serum sCD30 testing has been proposed in immunological risk estimation together with anti-HLA antibodies. We evaluated the risks associated with high pretransplant serum sCD30 in well HLA-matched cadaveric kidney recipients recruited in a clinical study comparing different immunosuppressive regimens. Rejection rate was similar in 37 recipients with high pretransplant serum sCD30 compared to 117 recipients with low serum sCD30 (16% vs. 15%, P=NS). Compared to pretransplant levels, the posttransplant sCD30 levels generally decreased, also in patients with rejection, although on day 21 posttransplant, rejecting patients had significantly higher relative sCD30 than nonrejecting patients (P<0.01). However, steroid-resistant rejection was associated with increasing posttransplant sCD30 levels. High pretransplant sCD30 values were associated with tubulointerstitial rejection. There was no correlation of sCD30 with delayed graft function. Good HLA matching seems to be effective in neutralizing the negative effect of a high pretransplant serum sCD30. PMID- 16378079 TI - The immunosuppressive agent FK506 enhances the cytolytic activity of inhibitory natural killer cell receptor (CD94/NKG2A)-expressing CD8 T cells. AB - Tacrolimus (FK506) is a potent immunosuppressive agent that inhibit transcription of cytokines such as IL-2 in T cells. The C-type lectin superfamily inhibitory NKR CD94/NKG2A-expressing cells can monitor the global status of HLA class I on tumor and leukemic cells through the recognition of HLA-E and induce cytolytic attack without an inhibitory signal against HLA class I-decreased target cells. We found that there was no effect of FK506 on the expansion of inhibitory NKR CD94/NKG2A-expressing T cells from granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood mononuclear cells (G-PBMCs). However, cytolytic activity levels of purified CD94-expressing cells from 7-day cultures with FK506 were much higher than those from 7-day cultures without FK506. These data suggested that FK506 did not inhibit cytolytic activities of inhibitory NKR expressing T cells and that there was a possibility of cytolytic activities being enhanced through the induction of cytotoxic molecules such as NKG2D and granzyme during a seven-day culture with FK506. PMID- 16378080 TI - Sirolimus use and de novo minimal change nephropathy following renal transplantation. PMID- 16378081 TI - Liver transplantation and hepatitis C virus. PMID- 16378082 TI - Opening the conversation. PMID- 16378085 TI - Monogamy: dopamine ties the knot. PMID- 16378086 TI - Internalizing channels: a mechanism to control pain? PMID- 16378087 TI - A functional genomics guide to the galaxy of neuronal cell types. PMID- 16378088 TI - Damaging secretions: chromogranins team up with mutant SOD1. PMID- 16378090 TI - MC4R neurons weigh in differently. PMID- 16378095 TI - A genetic strategy to treat sickle cell anemia by coregulating globin transgene expression and RNA interference. AB - The application of RNA interference (RNAi) to stem cell-based therapies will require highly specific and lineage-restricted gene silencing. Here we show the feasibility and therapeutic potential of coregulating transgene expression and RNAi in hematopoietic stem cells. We encoded promoterless small-hairpin RNA (shRNA) within the intron of a recombinant gamma-globin gene. Expression of both gamma-globin and the lariat-embedded small interfering RNA (siRNA) was induced upon erythroid differentiation, specifically downregulating the targeted gene in tissue- and differentiation stage-specific fashion. The position of the shRNA within the intron was critical to concurrently achieve high-level transgene expression, effective siRNA generation and minimal interferon induction. Lentiviral transduction of CD34(+) cells from patients with sickle cell anemia led to erythroid-specific expression of the gamma-globin transgene and concomitant reduction of endogenous beta(S) transcripts, thus providing proof of principle for therapeutic strategies that require synergistic gene addition and gene silencing in stem cell progeny. PMID- 16378096 TI - Association of beta-arrestin and TRAF6 negatively regulates Toll-like receptor interleukin 1 receptor signaling. AB - Tumor necrosis factor receptor-associated factor 6 (TRAF6) is critical for mediating Toll-like receptor (TLR)-interleukin 1 receptor (IL-1R) signaling and subsequent activation of NF-kappaB and AP-1, transcriptional activators of innate immunity. Here we show that beta-arrestins, a family of multifunctional proteins, directly interacted with TRAF6 after TLR-IL-1R activation. Formation of the beta arrestin-TRAF6 complex prevented autoubiquitination of TRAF6 and activation of NF kappaB and AP-1. Endotoxin-treated beta-arrestin 2-deficient mice had higher expression of proinflammatory cytokines and were more susceptible to endotoxic shock. Thus, beta-arrestins are essential negative regulators of innate immune activation via TLR-IL-1R signaling. PMID- 16378097 TI - CD45 links the B cell receptor with cell survival and is required for the persistence of germinal centers. AB - To segregate the many contributions that B cell receptor (BCR)-mediated signals make to immune responses, we have analyzed here B cells deficient in the 'pan leukocyte' marker CD45. BCR ligation of Cd45-/- B cells failed to activate phosphatidylinositol-3-OH kinase, NF-kappaB, Erk1 or Erk2 kinases or to upregulate cell survival proteins and instead induced apoptosis. Immunization of Cd45-/- B cell chimeras induced germinal centers and antigen-specific immunoglobulin G1 antibody-forming cells early, but both cellular compartments decreased by day 14. Proliferation of Cd45-/- B cells induced by CD40 ligand in vitro was impaired as a result of abrogation by BCR ligation of the upregulation of prosurvival proteins. In contrast, enforced expression of the antiapoptotic factor Bcl-xL prevented the collapse of Cd45-/- B cell germinal centers. These results show mechanistic differences in B cell survival during germinal center initiation and propagation; CD40 signaling is sufficient for the former, whereas the latter requires signaling from the BCR. PMID- 16378098 TI - Complex formation of Plk1 and INCENP required for metaphase-anaphase transition. AB - Mitotic chromosomal dynamics is regulated by the coordinated activities of many mitotic kinases, such as cyclin-dependent kinase 1 (Cdk1), Aurora-B or Polo-like kinase 1 (Plk1), but the mechanisms of their coordination remain unknown. Here, we report that Cdk1 phosphorylates Thr 59 and Thr 388 on inner centromere protein (INCENP), which regulates the localization and kinase activity of Aurora-B from prophase to metaphase. INCENP depletion disrupts Plk1 localization specifically at the kinetochore. This phenotype is rescued by the exogenous expression of INCENP wild type and INCENP mutated at Thr 59 to Ala (T59A), but not at Thr 388 to Ala (T388A). The replacement of endogenous INCENP with T388A resulted in the delay of progression from metaphase to anaphase. We propose that INCENP phosphorylation by Cdk1 is necessary for the recruitment of Plk1 to the kinetochore, and that the complex formation of Plk1 and Aurora-B on INCENP may play crucial roles in the regulation of chromosomal dynamics. PMID- 16378099 TI - GCP-WD is a gamma-tubulin targeting factor required for centrosomal and chromatin mediated microtubule nucleation. AB - The gamma-tubulin ring complex (gammaTuRC) is a large multi-protein complex that is required for microtubule nucleation from the centrosome. Here, we show that the GCP-WD protein (originally named NEDD1) is the orthologue of the Drosophila Dgrip71WD protein, and is a subunit of the human gammaTuRC. GCP-WD has the properties of an attachment factor for the gammaTuRC: depletion or inhibition of GCP-WD results in loss of the gammaTuRC from the centrosome, abolishing centrosomal microtubule nucleation, although the gammaTuRC is intact and able to bind to microtubules. GCP-WD depletion also blocks mitotic chromatin-mediated microtubule nucleation, resulting in failure of spindle assembly. Mitotic phosphorylation of GCP-WD is required for association of gamma-tubulin with the spindle, separately from association with the centrosome. Our results indicate that GCP-WD broadly mediates targeting of the gammaTuRC to sites of microtubule nucleation and to the mitotic spindle, which is essential for spindle formation. PMID- 16378100 TI - GENERATIVE CELL SPECIFIC 1 is essential for angiosperm fertilization. AB - The double fertilization process in angiosperms is based on the delivery of a pair of sperm cells by the pollen tube (the male gametophyte), which elongates towards an embryo sac (the female gametophyte) enclosing an egg and a central cell. Several studies have described the mechanisms of gametophyte interaction, and also the fertilization process - from pollination to pollen tube acceptance. However, the mechanisms of gamete interaction are not fully understood. Cytological studies have shown that male gametes possess distinct cell-surface structures and genes specific to male gametes have been detected in cDNA libraries. Thus, studies of isolated gametes may offer clues to understanding the sperm-egg interaction. In this study, we identified a novel protein, designated GCS1 (GENERATIVE CELL SPECIFIC 1), using generative cells isolated from Lilium longiflorum pollen. GCS1 possesses a carboxy-terminal transmembrane domain, and homologues are present in various species, including non-angiosperms. Immunological assays indicate that GCS1 is accumulated during late gametogenesis and is localized on the plasma membrane of generative cells. In addition, Arabidopsis thaliana GCS1 mutant gametes fail to fuse, resulting in male sterility and suggesting that GCS1 is a critical fertilization factor in angiosperms. PMID- 16378102 TI - [Estimation of an NY-ESO-1 expressing HCC cell line by NY-ESO-1b specific CD8+T cells in vitro induced by HLA-A2 restricted NY-ESO-1b peptide]. AB - OBJECTIVE: To analyze the effect of an NY-ESO-1 expressing HCC cell line by NY ESO-1b specific CD8(+)T cells in vitro induced by HLA-A2 Restricted NY-ESO-1b peptide and to further analyze the possibility of NYjESOj1 as a vaccine in HCC treatment. METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from an HLA-A2(+) HCC patient and NY-ESO-1 specific CD8(+)T cells were in vitro induced by HLA-A2 Restricted NY-ESO-1b peptide. Next we estimated the effect that NY-ESO-1 specific CD8(+)T cells recognized and killed NY-ESO-1 expressing HepG2 cells by enzyme-linked immunospot(ELISPOT). RESULTS: These in vitro induced NY ESO-1b specific CD8(+)T cells could recognize HepG2 cells stably transfected with NY-ESO-1 in both IFN-gamma and Granzyme B ELISPOT assays. And the data also showed that NY-ESO-1 specific CD8(+)T cells could efficiently kill NY-ESO-1 expressing HepG2 cells. CONCLUSION: NY-ESO-1 protein can be processed by HCC cells, and that HLA-A2 restricted NY-ESO-1b peptide (157-165) can also be presented on the surfaces of cells and recognized by NY-ESO-1b specific CD8(+)T cells in vitro induced. PMID- 16378103 TI - [Comparison of the immunomodulatory effects of spore polysaccharides and broken spore polysaccharides isolated from Ganoderma lucidum on murine splenic lymphocytes and peritoneal macrophages in vitro]. AB - OBJECTIVE: To compare the immunomodulatory effects of spore polysaccharides (Gl SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. METHODS: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-gamma and TNF alpha in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. RESULTS: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-alpha and NO in Gl-SP- or Gl BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8 mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-gamma in doublejway MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2 12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4(+) or CD8(+) subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4(+) and CD8(+) subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. CONCLUSION: Gl-SP and Gl BSP have similar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP. PMID- 16378104 TI - [Establishment of a stable and regulable 293 cell line expressing mycoplasma hyorhinis protein P37]. AB - OBJECTIVE: To establish a stable cell line, which can express P37 protein of mycoplasma hyorhinis and be regulated by tetracycline, for investigating the effect of p37 on phenotype of cells and its mechanism. METHODS: Recombinant plasmid PcDNA5/FRT/TO-p37 was constructed and cotransfected with pOG44 into Flp In-T-REx-293 cells by lipofectamine. Positive clones were screened with Hygromycin and Blasticidin. RT-PCR and Western blot were used to exam the mRNA and protein expression in selected clones. The expression level at different inducing times and concentrations of tetracycline were examined. MTT assay was used to observe the effect of P37 on proliferation of 293 cells. RESULTS: P37 protein, which is 43.5x10(3), was expressed in the selected clone as well as secreted from cells. Tetracycline showed a good regulation on the expression of P37 protein, which was not detectable without tetracycline induction. When induced with 2 mg/L tetracycline for 60 hours, the P37 protein expression reached maximum level. Cell growth was promoted after being transfected with p37. CONCLUSION: A stable cell line expressing P37 regularly was established, which provides a good cell model for studying p37 function and its molecular mechanism. PMID- 16378105 TI - [Alteration of hydrogen sulfide/cystathionine-beta-synthase system in rats with recurrent febrile seizures]. AB - OBJECTIVE: To study the alteration of hydrogen sulfide (H(2)S)/ cystathionine beta-synthase (CBS) system during recurrent febrile seizures (FS) in the hippocampus of developing rats. METHODS: The rats were randomly divided into control group (n=8) and hyperthermia-treated group (n=22). Which was subdivided into FS group (n=8) and H group(no seizure occurred, n=9) according to whether seizures occurred. The plasma level of H(2)S was detected by the spectrophotometer. The expression levels of CBS gene and protein were examined by in situ hybridization and immunohistochemistry respectively. RESULTS: The plasma levels of H(2)S were increased significantly in FS group compared with those of control group or H group. The expression levels of CBS gene and protein were enhanced in FS group compared with those of control group or H group. CONCLUSION: The expression levels of H(2)S/ CBS system were up-regulated during recurrent FS. PMID- 16378106 TI - [Relationship between HLA-DR4/1 subtypes and T cell responses to type II collagen or anti-C II 263-272 antibodies]. AB - OBJECTIVE: To investigate the relationship between HLA-DR4/1 subtypes and T cell responses to type II collagen or anti-CII 263-272 antibodies. METHODS: Peripheral blood mononuclear cells and sera were obtained from 70 rheumatoid arthritis (RA) patients. T cell proliferative responses to CII 263-272 were evaluated by [(3)H] thymidine incorporation assay. Antibodies specific to CII 263-272 were determined by ELISA. HLA-DR4/1 subtypes were analyzed by PCR-SSP. RESULTS: In HLA-DR4/1 positive group, T cell proliferative responses to CII 263-272 were observed in 55.6% RA patients and anti-CII 263-272 antibodies were present in 66.7% RA patients. In HLA-DR4/1 negative group, T cell proliferative responses to CII 263 272 were observed in 30.3% RA patients and anti-C II 263-272 antibodies were present in 34.8% RA patients. The number of positive antibodies to CII 263-272 or T cell proliferative responses to CII 263-272 in HLA-DR4/1 positive group were higher than in negative group. CONCLUSION: There is a relationship between HLA DR4/1 subtypes and T cell responses to CII 263-272 or anti-CII 263-272 antibodies. PMID- 16378108 TI - [Contribution of MODY2 gene to the pathogenesis of Chinese early onset familial type 2 diabetes]. AB - OBJECTIVE: To assess the prevalences of maturity onset diabetes of the young 2 (MODY2) in Chinese early onset familial type 2 diabetic population and explore the possibility of some common DNA variants of MODY2 gene to contribute to the susceptibility of type 2 diabetes in Chinese population. METHODS: A total of 100 early onset type 2 diabetes pedigrees in Beijing were collected. By PCR, all the exons and exon/intron splice sites of MODY2 gene were amplified, and PCR products were sequenced to identify the DNA variants . The SNPs were genotyped by homogenous mass extend (hME) assay in nonjdiabetic control population. RESULTS: Three DNA vatiants in MODY2 gene were identified: one in exon4(ccc-->ccg, heterozygous silent mutation,pro145pro), one in exon7(gcc-->gcg, heterozygous silent mutation, ala233ala), and one in intron 9 (IVS9+8 C>T). The prevalences of these variant alleles were 0.5 in exon 4, 0.5% in exon 7 and 36% in intron 9. An association study was conducted to examine the relationship between the IVS9+8 C>T polymorphism and early onset familial type 2 diabetes, the frequency of allele T of early onset familial type 2 diabetes population was significantly lower, and the frequency of allele C significantly higher, than those of control subjects (P<0.05). CONCLUSION: Our research suggested in Chinese population the prevalences of MODY2 was less than 1% in early onset familial type 2 diabetic patients, the IVS9+8 C>T polymorphism at MODY2 gene, intron 9 or nearby genes was associated with early onset familial type 2 diabetes . PMID- 16378107 TI - [Association and linkage analysis between the S447X polymorphism of LPL gene and serum lipids, blood pressures in twins]. AB - OBJECTIVE: To investigate the association and linkage between the S447X polymorphism of LPL gene and serum lipids, blood pressures in the general population-based twin cohort of China. METHODS: The twin subjects were collected based on the twin registry system of China. All twins were investigated by a standard questionnaire and physical examinations. PCR-RFLP method was used to detect the genotypes of S447X. Stratified by gender, the associations of S447X polymorphism with the serum lipids (TG, HDL), blood pressures (SBP, DBP) and their abnormalities were analyzed by univariate and multivariate methods. The Halesman-Elston regression based nonjparameter linkage analysis was applied in the DZ twins. RESULTS: A total of 962 adult twin pairs were analyzed and the frequency of 447X allele was 8.6%. The heritabilities of TG, HDL, SBP and DBP were 57%, 68%, 46% and 44%, respectively. In female twins, univariate and multivariate association analyses showed that the 447X allele was associated with the 12.9% decrease of TG and 2.7 mm Hg (1 mm Hg=0.133 kPa) decrease of SBP, 1.8 mm Hg decrease of DBP. Also in female twins, 447X allele was associated with the lower risks of high TG dyslipidaemia, low HDL dyslipidaemia and hypertension, with the odds ratios 0.38 (95%CI: 0.20-0.76), 0.49 (95%CI: 0.29-0.81) and 0.47(95%CI: 0.25-0.86), respectively. CONCLUSION: This study showed significant associations between the S447X polymorphism of LPL gene and the improved serum lipids, decreased blood pressures in the female twins. But no associations were observed in male twins, and no linkage evidences were found between the locus site of S447X polymorphism and the trait loci of serum lipids, blood pressures. So the associations maybe caused by the other functional SNPs within or near LPL gene, which might be in significant linkage disequilibrium with 447X mutation. PMID- 16378109 TI - [Regulation of expression of cGMP-dependent protein kinase Ialpha by calcineurin in vascular smooth muscle cells proliferation]. AB - OBJECTIVE: To discuss the regulation of expression of cGMP-dependent protein kinase I(alpha) (PKG I(alpha)) by calcineurin (CaN) in vascular smooth muscle cells (VSMCs) proliferation. METHODS: Cultured wistar rat aortic VSMCs were used as an experimental model. CaN was inhibited by its special inhibitor cyclosporin A (CsA). Phenylephrine (PE) was given to stimulate VSMCs to proliferate. All of cultured cells were divided into four groups:control group, 0.5 mg/L CsA group, 5 mg/L CsA group and 5 mg/L CsA+10 micromol/L PE group. The mRNA and protein expressions were assayed by RT-PCR, immunocytochemistry and Western blot analysis. RESULTS: The OD ratio of PKG I(alpha) mRNA expression in 0.5 mg/L CsA group resembled that in the control group while that in 5 mg/L CsA group was significantly higher than that in the control group (P<0.01). Although the result of 5 mg/L CsA+10 micromol/L PE group was lower than that in 5 mg/L CsA group (the expression of mRNA decreased 32.2%; the production of PKG I(alpha) protein decreased 36.7%, P<0.01), but still higher than that in the control group obviously (P<0.05). This kind of complexion was also validated by immunocytochemistry and Western blot analysis. CONCLUSION: CaN is a regulator of PKG I(alpha) expression in cultured VSMCs proliferation. Once CaN was inhibited by its special inhibitor CsA, the expression of PKG I(alpha) increased obviously. And the expression reduced significantly when CaN was actived by PE again. PMID- 16378110 TI - [Cross-talk between calcineurin and protein kinases in airway remodeling in asthma]. AB - OBJECTIVE: To determine the role of cross-talk between calcineurin-dependent signal transduction pathway and protein kinase C (PKC), mitogen-activated protein kinase (MAPK) and protein kinase A (PKA) in airway remodeling in asthma. METHODS: Male guinea pigs were sensitized with intraperitoneal injections of ovabumin (OVA), then treated with cyclosporin A (CsA, 5 mg/kg), an inhibitor of calcineurin, then inhaled OVA for 2 weeks 14 days later. Activities of calcineurin, PKC, MAPK, and PKA were was analyzed by phosphorylation and dephosphorylation. In primary cultures of rat airway smooth muscle cells (ASMC), activities of calcineurin, PKC, MAPK, and cross-talk induced by urotensin II (UII ), a recently identified strong mitogen, were measured. RESULTS: (1) The activities of calcineurin, MAPK and PKC increased by 19% (P<0.01), 28% (P<0.01) and 35% (P<0.05), respectively, in the asthmatic group compared with controls but decreased by 52% (P<0.01), 18% (P<0.05) and 52% (P<0.01), respectively, in the CsA group compared with asthmatic group. PKA activity in the asthma group decreased by 53% (P<0.01) compared with controls but increased by 2.65-fold (P<0.01) in the CsA group compared with the asthma group. (2) UII 10(-7) mol/L stimulated ASMC PKC and MAPK activities by 44% and 24% (P<0.01), respectively, after incubating for 20 min. It increased CaN activity in a time-dependent manner, being 1.67 times that of the control for 24 h (P<0.01). (3) CsA 10(-6) mol/L and H(7) 50 micromol/L, an inhibitor of PKC, inhibited UII-stimulated CaN activity by 45% (P<0.01) and 21% (P<0.05), respectively, while PD(98059) 50 micromol/L, an inhibitor of MAPK, had no effect on CaN activity (P>0.05). (4) CsA 10(-6) mol/L inhibited UII-stimulated PKC activity by 14% (P<0.05), while having no effect on MAPK activity (P>0.05). CONCLUSION: The signal transduction pathways between calcineurin and other protein kinases such as PKC, MAPK and PKA have cross-talk in airway remodeling in asthma. PMID- 16378111 TI - [Lysophosphatidic acid activates L-arginine/nitric oxide pathway of platelets in rats]. AB - OBJECTIVE: To investigate the mechanism of platelet function caused by Lysophosphatidic acid (LPA), by observing the change of the L-arginine/nitric oxide synthase/nitric oxide (L-Arg/NOS/NO) pathway of platelet in rats. METHODS: LPA (10(-6), 10(-5) and 5x10(-5) mol/L) was administrated in rats and incubated for 30 and 60 minutes. The nitrite production was measured by Greiss assay; NOS activities and L-arginine transportation were detected by isotope tracer method and intracellular [Ca(2+)]i changes by fluorescent probe. RESULTS: LPA increased NO release by 35% and 56%, after incubating for 30 and 60 minutes, respectively. LPA (10(-6), 10(-5)aand 5x10(-5) mol/L) enhanced the NO productions of platelets in a concentration-dependent manner (P<0.01). EC(50) was 17.8 micromol/L, and 95% CI was 13.3-24.2 micromol/L, involved in the physiological concentration of LPA in plasma (P<0.01). Simultaneously, different doses of LPA increased NOS activities and L-arginine uptake in a dose-dependent manner (P<0.01). In this study, LPA (50 micromol/L) increased the intracellular free calcium ion concentration ([Ca(2+)]i, P<0.01), after incubating for 30 and 60 minutes. Pre treated with NOS inhibitor-L-NAME for 20 minutes, LPA obviously enhanced the effects by 20% and 32% respectively (P<0.01). On the contrary, pre-treated with L arginine (200 micromol/L) for the same times obviously reduced the effects by 14% and 18% respectively (P<0.01). CONCLUSION: LPA increased NO release by enhancing L-arginine uptake and NOS activities, up-regulating L-arginine/NOS/NO pathway in platelets of rats. PMID- 16378112 TI - [Cell-cycle negative regulatory gene ANA is over-expressed in the brain tissues of patients with Down syndrome]. AB - OBJECTIVE: To investigate the expression level of genes located in chromosome 21 in the brain tissues of Down syndrome(DS). METHODS: An optimized semi quantitative RT-PCR method was used to evaluate the expression levels of seven genes encoded in chromosome 21 in fetal cortex brain and cerebellum of DS and the control at the end of 20 weeks of gestation. B2M was used as internal reference to normalize cell loss. RESULTS: The expression levels of 6 genes in cortex and cerebellum, including DYRK1A, SYNJ1, PCP4, C21orf5, C21orf2 and C21orf106, were comparable between DS and the control. ANA, a cell-cycle negative regulatory gene, was over-expressed dramatically in the cortex but not in the cerebellum of DS. CONCLUSION: Over-expression of ANA may contribute to the reduction of neuronal density in DS brain. PMID- 16378114 TI - [Effects of lidocaine on impairment of learning and memory function and cholinergic system caused by cerebral microsphere embolism in rats]. AB - OBJECTIVE: To investigate the effects of lidocaine on impairment of learning and memory function and cholinergic system caused by cerebral microsphere embolism in rats. METHODS: Healthy male Wister rats were randomly divided into the following groups. (1) Control group. (2) 600 microsphere group and 900 microsphere group, in which 600 or 900 microspheres were injected into the right internal carotid artery, respectively. (3) 600 treatment group and 900 treatment group, in which 600 or 900 microspheres were injected into the right internal carotid artery, respectively, and lidocaine was given. Water maze tasks were tested for 5 consecutive days from the 7th postoperative day. The rats were then decapitated and regions of cerebral cortex, hippocampus, and striatum were selected. The activities of choline acetyltransferase and cholinesterase and the binding activity of muscarinic receptor were determined. RESULTS: (1) The latency periods were significantly longer in the 900 microsphere group than in the control group and in the 600 microsphere group. (2) The percentages of effective search strategy were significantly lower in the 600 and 900 microsphere groups than in the control group. They were significantly higher in the 600 and 900 treatment groups than in the corresponding microsphere groups. (3) The activities of choline acetyltransferase of cerebral cortex were significantly lower in the 900 microsphere and two treatment groups than in the control group. They were also significantly lower in the 600 and 900 treatment groups than in the corresponding microsphere groups. Those of striatum were all significantly lower in the microsphere and treatment groups than in the control group. (4) The activities of cholinesterase of cerebral cortex were significantly lower in the 900 microsphere group than in the control and 600 microsphere groups. They were significantly higher in the 900 treatment group than in the 900 microsphere group. Those of hippocampus were all significantly lower in the microsphere and treatment groups than in the control group. (5) The binding activities of muscarinic receptor of cerebral cortex were significantly lower in the 900 microsphere and two treatment groups than in the control group. They were also significantly lower in the two treatment groups than in the corresponding microsphere groups. Those of hippocampus and striatum were all significantly lower in the microsphere and treatment groups then in the control group. They were also significantly lower in the 600 or 900 treatment group than in the corresponding microsphere group. CONCLUSION: Cerebral microsphere embolism caused significant and quantity dependent impairment of learning and memory function and cholinergic system in rats. Lidocaine alleviated learning and memory dysfunction caused by cerebral microsphere embolism, but further inhibited the parameters of central cholinergic system. PMID- 16378113 TI - [Disease transformation in imatinib mesylate treated Philadelphia chromosome positive chronic myelogenous leukemia patients achieved cytogenetic remissions]. AB - OBJECTIVE: To report disease transformation in 3 imatinib mesylate treated Philadelphia chromosome-positive chronic myelogenous leukemia (Ph(+)CML) patients who achieved cytogenetic response. METHODS: Hematologic examinations and bone marrow G-banding karyotyping were evaluated at regular intervals in 3 patients with Ph(+)CML who achieved hematologic responses during continuous imatinib therapy, including 1 patient in first chronic phase (case 1), 1 patient in second chronic phase (case 2) and 1 patient in accelerated phase (case 3). RESULTS: Case 1, case 2 and case 3 achieved complete cytogenetic response after 4, 3 and 6 months of imatinib therapy respectively. Though under continuously imatinib treatment, they developed acute lymphoblastic leukemia, acute myelogenous leukemia and extramedullary blast crisis in the following 12, 6 and 0 months respectively. Cytogenetic studies of bone marrow in the crisis period showed that cells in case 1 and case 3 had a complete cytogenetic response (Ph(+)cell=0), but case 2 had 20% Ph(+) cells. CONCLUSION: Acute leukemia or extramedullary blast crisis with Ph(-) cells or dominant Ph(-) cells in bone marrow may occur in the patients with Ph(+)CML after imatinib therapy. PMID- 16378115 TI - [Influence of lidocaine on systemic inflammation in perioperative patients undergoing cardiopulmonary bypass]. AB - OBJECTIVE: To investigate the influence of lidocaine on systemic inflammation in the perioperative ventricular septal defect (VSD). METHODS: Twenty patients, scheduled for ventricular septal defect were randomly divided into 2 groups: lidocaine and control groups. Before rebeat lidocaine 1 mg/kg was given. The venous blood samples were obtained from the central venous at the following points: after induction of anesthesia and before cardiopulmonary bypass(CPB,T1),1 h after CPB(T2),2 h after CPB(T3), and 4 h after CPB(T4). IL-6 and IL-8 were determined by radio-immunoassay. RESULTS: Compared with those at T1, the levels of white blood cells,polymorphonuclear neutrophils,IL-6 and IL-8 increased significantly from T2 to T4 in both groups. IL-6 and IL-8 levels reached the peak at T2. Compared with those in control groups, IL-6 level decreased obviously in lidocaine group from T2 to T4, but IL-8 level remained unchanged significantly. CONCLUSION: Under CPB and VSD repair the systemic inflammation is obvious, reaches the peak 30 min after CPB and persists to 4 h after CPB. Perioperative administration of lidocaine is effective against the inflammation. PMID- 16378116 TI - [Role of serum leptin and tumor necrosis factor-alpha in malnutrition of male chronic obstructive pulmonary disease patients]. AB - OBJECTIVE: To explore the function of serum leptin in COPD patients with malnutrition, and to investigate the relationship between leptin and TNF-alpha. METHODS: A total of 81 subjects (47 COPD patients and 34 control subjects) participated in this study. The 47 COPD patients were divided into 2 groups: group COPD I (patients without malnutrition during stable disease, n=29), group COPDII (patients with malnutrition during stable disease, n=18). To eliminate the effect of sex differences, all the patients and controls were male. Body mass index (BMI), percent ideal body weight (IBWa), triceps skin-fold thickness (TSF), mid-upper arm circumference (MAC), mid-upper arm muscle circumference(MAMC),serum leptin and tumor necrosis factor-alpha(TNF-alpha) levels, serum prealbumin (PA), serum transferrin (TF), serum albumin(Alb),total lymphocytes count (TLC), forced expiratory volume in one second (FEV(1)), maximal inspiration pressure(MIP)and maximal expiration pressure(MEP)were measured in all participants. Leptin levels were measured by radioimmunoassay. TNF-alpha levels were measured by ELISA. The between group difference and correlation of these parameters were analysed. RESULTS: (1) Serum leptin levels were significantly lower in group COPDII (4.07+/ 3.42 microg/L) than in group COPD I (9.72+/-6.67 microg/L) and controls (8.21+/- 5.41 microg/L, P<0.05). (2) There were no statistical differences in serum TNF alpha levels between group COPDII (8.03+/-3.37 ng/L)and group COPD I (7.25+/- 2.08 ng/L). (3) There was no significant correlation between leptin and TNF-alpha in any group. CONCLUSION: Leptin was not involved in anorexia and weight loss of COPD patients. There was no significant correlation between TNF-alpha and leptin during the regulation of the energy balance in COPD patients. PMID- 16378117 TI - [Duraplasty with Neuropatch versus autologous fascia lata for Chiari I malformation with syringomyelia: a comparative study]. AB - OBJECTIVE: To evaluate the outcome and postoperative reaction of dural substitute (Neuropatch) applying in the treatment of Chiari I malformation(CMI) associated with syringomyelia(SM). METHODS: Forty patients of CMI associated with SM were operated in our department from Jul. 2002 to Jul. 2004. All patients underwent posterior cranial fossa decompression and duraplasty. They were divided into two groups, 20 patients being repaired with Neuropatch (Neuropatch group), and the others with autologous fascia lata (fascia group). There were 6 males and 14 females in Neuropatch group and 10 males and 10 females in fascia group. The operations were performed under general anesthesia via suboccipital approach and the extent of posterior cranial fossa decompression ranged from 20 cm(2) (5 cmx4 cm) to 35 cm(2) (5 cmx7 cm). The removal of posterior arch of atlas depended on the extent of tonsillar herniation, and the dura was opened in Y shape. The Neuropatch was cut into triangular shape, and the same sized autologous fascia lata was used in fascia group. The patches were sutured tightly to the dura matter in each group. The incision was closed layer by layer and drainage was used, if necessary. Antibiotics and hormone were routinely used. The duration of operation, postoperative fever were evaluated, the outcome of the operation was evaluated by Tator scale, and the data were analyzed with statistic software SPSS 10.0. RESULTS: There were12 patients (60%) who suffered from postoperative fever in the Neuropatch group, and 9 patients (45%) in the fascia group(chi(2)=0.902,P=0.342). Seventeen patients in each group were improved postoperatively. The duration of operation, postoperative fever and antibiotics used were compared between the two groups. No significant difference was found, but the duration of postoperative fever and the time of hormone used were different. There were no postoperative infections that occurred after the follow up for 1 to 2 years, except for one patient in fascia group who developed infective granuloma and recovered later by treatment. CONCLUSION: Neuropatch is a useful dural substitute for the repair of dural defects in the treatment of CMI associated with syringomyelia. PMID- 16378118 TI - [Comparison of single plasma sample methods and prediction of dual plasma sample method in measurement of 99mTc-Diethylene Triamine Pentaacetic Acid plasma clearance]. AB - OBJECTIVE: To find an applicable condition of the single-plasma-sample method (SPSM) to measure the glomerular filtration(GFR) with (99m)Tcj Diethylene Triamine Pentaacetic Acid ((99m)Tc-DTPA), and predict the value of (99m)Tc-DTPA plasma clearance by dual plasma sample method (DPSM ) from that by SPSM. METHODS: Three hundred and thirty five patients with chronic kidney disease (CKD) were selected (192 males and 143 females). The average age was 51.91+/-14.76 years. The GFR was determined simultaneously by 2 methods: (1) SPSM (sGFR); (2) DPSM (tGFR), using DPSM as reference standard, sGFR calculated from the different SPSM was compared with tGFR. An equation was developed to predict tGFR from sGFR. GFR estimated by abbreviated Modification of Diet in Renal Disease (MDRD) equation (aGFR) was evaluated as the criterion in selection of DPSM and SPSM. The condition that DPSM could be substituted by SPSM in GFR measurement was given. RESULTS: When tGFR >or= 30 mL/(minx1.73 m(2)), all of the sGFR were significantly correlated with tGFR. Among them, Watson modified Christensen and Groth's equation at sample time=240 min tended to be the most accurate (r=0.977, RMSE=10.91), and tGFR could be predicted from sGFR using the equation: Predicted tGFR [mL/(minx1.73 m(2))]=7.755 4+0.789 3xsGFR+0.002 4 xsGFR2 (n=297, r2=0.959 1, P<0.001). When aGFR was less than 50 mL/(minx1.73 m(2)), the diagnostic sensitivity of tGFR<30 mL/(minx1.73 m(2)) was 94.7%, and recommended as the criterion in selection of DPSM and SPSM. CONCLUSION: When aGFR >or=50 mL/(minx1.73 m(2)), DPSM can be substituted by SPSM in GFR measurement. PMID- 16378119 TI - [Regulation of expression of pituitary tumor transforming gene 1 (PTTG1) by androgen in prostate cancer]. AB - OBJECTIVE: To identify the androgen-responsive genes in prostate and screen the molecular targets for further studying human prostate cancer. METHODS: The potential androgen-responsive gene pituitary tumor transforming gene 1 (PTTG1) was selected which had been previously screened by cDNA microarray in rat prostate and its mRNA level was detected by Northern blot in the castrated rat prostate with and without replacement of Mibolerone. Immunohistochemistry was performed to determine the expression and location of PTTG1 in human prostate tissues. Then human androgen-dependent prostate cancer cells LNCaP were used as a model to study the regulation of PTTG1 by Mibolerone. RESULTS: PTTG1 mRNA was hardly detectable in the prostate of 7-day castrated rats, while it was up regulated dramatically in the prostate of 7-day castrated rats treated with Mibolerone for 2 days. It was showed that high expression of PTTG1 was localized to the epithelial cells of human prostate cancer but not to the stromal cells with Immunohistochemistry. Northern blot analysis indicated that LNCaP cells treated with 0.1 nmol/L Mibolerone for 2 days led to the high PTTG1 mRNA expression. The basic expression of PTTG1 in human androgen-independent prostate cancer cell lines PC3 or DU145 was even higher than that in the human androgen dependent prostate cancer cells LNCaP treated with Mibolerone. CONCLUSION: Androgen can up-regulate the PTTG1 expression in castrated rat prostate and human prostate cancer cell LNCaP. It suggests that PTTG1 is potential to play an important role in human prostate cancer progression. PMID- 16378120 TI - [Withdrawal symptoms of ethanol are ameliorated by thymopentin in mice]. AB - OBJECTIVE: To explore the effect of thymopentin (TP5) on the choice of ethanol and ameliorating withdrawal symptoms (anxiety) of ethanol in mice. METHODS: Mice were administered ethanol (v/v) in schedular fashion: 5% (1 week), 10% (1 week) and 15% (4 weeks), followed with the free choice between ethanol and water. Ethanol/(ethanol+water)x100% [E/(E+W)x100%] was measured as an index of ethanol selection. Light-dark box test and elevated plus maze test were chosen for the assessment of anxiety pre-drug and post-drug. After TP5 [0.2 mg/(kgxd), 0.4 mg/(kgxd)], i.p. or saline (vehicle control), i.p. for 14 days, the procedure was repeated. RESULT: (1) E/(E+W)x100%: the post-drug values of TP5 (0.2 mg/kg, 0.4 mg/kg) were lower significantly than the pre-drug values. (2) Light-dark box test: the post-drug values of number of entries and time spent in the light chamber of TP5 (0.2 mg/kg, 0.4 mg/kg) were more than the pre-drug values themselves and the post-drug value of saline. (3) Elevated plus maze test: the post-drug values of time spent on open arms of TP5 (0.2 mg/kg, 0.4 mg/kg) were more than the pre-drug values themselves and the post-drug value of salineiand the post-drug values of time spent on close arms of TP5 were less than the pre drug values. CONCLUSION: TP5 could decrease the uptake of ethanol and ameliorate anxious behavior associated with ethanol withdrawal in mice. PMID- 16378121 TI - [Preoperative diagnosis and microsurgery of the optic gliomas:retrospective analysis of 7 patients]. AB - OBJECTIVE: To summarize the clinical characteristics and therapeutic experience of optic gliomas. METHODS: The clinical data of 7 patients of our department in the past 3 years were analyzed. RESULTS: In this series of optic glioma patients, the average age was 18.7 years, the ratio of men to women was 4:3, and 100% of the tumors were sub-totally removed. Diabetes inspidus was the most comman postoperative complication. CONCLUSION: The optic glioma is benign intracranial tumor with good prognosis. The key point of treatment is surgical resection combined with proper postoperative radiotherapy. PMID- 16378122 TI - [Effects of maxillary distraction osteogenesis on the velopharyngeal configuration of cleft palate patients]. AB - OBJECTIVE: To study the effects of internal maxillary distraction osteogenesis(DO) on the velopharyngeal configuration of cleft palate patients. METHODS: Ten patients with severe maxillary hypoplasia secondary to cleft lip and palate patients (7 males and 3 females, average age 20.1 years old) had undertaken high step LeFort I osteotomy, and internal maxillary distraction devices were applied to advance the maxilla. Before surgery, when DO was completed and 6 months after DO was completed, oriented lateral cephalograms at rest position of each patient were taken, and 6 measure indexes of velopharyngeal configuration were collected and analyzed. RESULTS: All patients had successfully accomplished maxillary DO and the maxilla had been averagely advanced 11.3 mm. PNS-PhW, C-PhW, UL and ANS-PNS-T had all significantly increased, and UD had significantly decreased when DO was completed and 6 months after DO was completed as compared with pre-surgery. No significant linear correlation was found between maxilla advancement distance and velopharyngeal configuration changes. CONCLUSION: Correction of maxillary hypoplasia secondary to cleft palate surgery by using internal maxillary DO can increase the velopharyngeal cavity depth, and may impair velopharyngeal competence, but the compensatory changes of velopharyngeal soft tissue can alleviate this impairment to certain extent. PMID- 16378123 TI - [A pilot study on the effects of Mini-SG F attachments on periodontal health of the abutment teeth used in free-end partial removable dentures]. AB - OBJECTIVE: To investigate the effects of Mini-SG F attachment on periodontal health of the abutment teeth in restoring free-end removable partial dentures (RPDs). METHODS: The periodontal status of the abutments in restoring free-end RPDs was longitudinally observed with periodontal indices commonly used in clinics, and a group comparison was made using a one-way analysis of variance (ANOVA). RESULTS: The stability and retention of free-end RPDs retained by Mini SG F attachments were excellent during the whole observation course. Compared with the periodontal status of the first and the second abutments before restoration respectively, no significant differences in gingival indices, plaque indices, periodontal probing depth, alveolar bone resorption levels of those were shown 6 months after restoration. The mobility of the abutments was significantly reduced when they were splinted together. CONCLUSION: Mini-SG F attachments used in free-end RPDs have no evident adverse effect on periodontal health of the abutment teeth, and the way of splinting the abutments is advantageous to their long-term health and stability. PMID- 16378124 TI - [Preparation of vascular calcification in vivo and vascular smooth muscle cell calcification in vitro of rats]. AB - OBJECTIVE: To investigate simple, economical, stable and efficient methods of in vivo and in vitro cardiovascular calcification models in rats. METHODS: Rats received Vitamin D(3) (300 000 U/kg, i.m.) and nicotine (25 mg/kg, 5 mL/kg in peanut oil, p.o.) at 8 a.m. on day 1. The nicotine administration was repeated at 6 p.m. Rats in control group received an injection of normal saline (i.m.) and two gavages of medium oil. All of the rats were then allowed to recover for 4 weeks and given standard rodent chow. After the measurement of cardiac function the rats were sacrificed and the calcium content in myocardium and aorta were measured. Von Kossa staining was used to detect the deposit of calcium in myocardium and aorta. Cultured smooth muscle cell(SMC) derived from rat thoracic aorta was treated with beta-glycerophosphate for 14 days, then the calcium content and deposit were measured. RESULTS: Compared with control group, the rats with cardiovascular calcification showed a lower body weight, The ratio of left heart to body weight, myocardial and aortic calcium content were increased respectively. Alkaline phosphatases activity in calcified myocardium and aorta were increased respectively, compared with the control. The values of animal mean blood pressure (MBP), heart rate (HR) and left ventricle end-distolic pressure (LVEDP) showed no significant alteration (P>0.05) in vitamin D3 plus nicotine (VDN) group. The values of +LV dp/dt(max) and -LV dp/dtmax were significantly lower in VDN group (P<0.05 and P<0.01, respectively). In calcified vascular smooth muscle cells(VSMCs), von Kossa staining for calcification, showed positive staining as black/brown areas within the main, large, nodular structures as shown in extracellular matrix and cytoplasma. The content of calcium, (45)Ca(2+) uptake and alkaline phosphatase (ALP) activity in calcified VSMCs were increased (all P<0.01), respectively, compared with that of the control. CONCLUSION: These methods can be used to produce calcification models in vivo and in vitro, which save money and time and are easy to manipulate. PMID- 16378125 TI - [Diffuse muscular calcification with subcutaneous cysts]. AB - OBJECTIVE: Diffuse muscular calcification was rare myopathological change due to abnormal metabolism of calcium, which was mainly found in dermatomyositis and myositis ossificans progressiva. Here we reported a case of diffuse muscular calcification that clinically mimicked myositis ossificans progressiva. The disease might be a new type of congenital calcium metabolic disease. METHODS: A 15-year-old girl developed subcutaneous cysts in the wrist and ankle when she was 1 year old. At the age of 9, she developed recurrent fever with myalgia, fatigue and diffuse muscular calcification. It was difficult for her to squat, run or walk. Protuberance presented in the subcutaneous tissue of her trunk. Some nodules ruptured with outflow of chalky material. ESR, ENA, RF, CRP, PTH, CK were in normal limits. EMG was unremarkable. X-ray confirmed diffuse calcification in the muscle and subcutaneous tissues. Biceps muscle biopsy was performed. RESULTS: Numerous inflammatory cells infiltrated around vessels in the perimyosium with perifascicular muscle fiber atrophy and degeneration. Many RRF and SDH positive fibers were also observed. EM showed tubular reticular inclusions in vascular endothelium. CONCLUSION: Diffuse muscular calcification indicated existence of systemic calcium metabolic abnormality. As the clinical symptoms and distribution pattern of calcification were different from dermatomyositis with subcutaneous calcification and myositis ossificans progressiva, our case might be a new type of disease. The microvascular changes might result in the lesion of muscle fibers. PMID- 16378126 TI - [Family of multiple peptide fragments derived from angiotensin and their interaction]. AB - It is well established that renin-angiotensin system (RAS) is the major regulatory network that maintains blood pressure, fluid and electrolyte balance and the homeostasis of cardiovascular system. Most studies in the last decades centered on the pivotal members of RAS, that is, angiotensin II (Ang II ) and angiotensin converting enzyme (ACE). Recent identification of ACE-2, the homology of ACE, and the identification of the multiple products degraded from angiotensin I, including Ang III, Ang IV, Ang(1-9), Ang(1-7), Des-Asp-angiotensin I(DAA I), etc, have proved that Ang II is not the only biological active compound of RAS. Peptides derived from angiotensinogen,that is, family of angiotensins,display independent and pleiotropic biological activities in vivo, and interact with each other both in metabolism pathway and the biologic effects. The imbalance of the network of angiotensin metabolites exhibits significant pathophysiological role in cardiovascular disease. PMID- 16378127 TI - [Short circuit current technology and its application in the study of traditional Chinese medicine]. AB - Short circuit current (I(sc)) technique has been applied in the studies of transepithelial ion transport in various epithelia. Recently it has also been used in the modernization researches on traditional Chinese medicines. This review gives an overview of the basic principle of the I(sc) technique, the targets of measurement, ion transport, ion channel, the general ways of I(sc) research design, the application of I(sc) technology in the researches on traditional Chinese medicines. PMID- 16378128 TI - [Long term (5 years) outcome of radiofrequency ablation for hepatocellular carcinoma in 256 cases]. AB - A total of 267 patients with hepatocellular carcinoma underwent ultrasound-guided radiofrequency ablation (RFA) in Peking University School of Oncology between 1999 and 2005 (421 RFA sessions). Among them, 254 patients were candidates for RFA treatment and the selective criteria were: (1) the greatest diameter of tumor 5 cm, respectively. According to tumor size, shape and location, we adopted a defined treatment strategy, which consisted of a mathematical protocol, an individualized protocol and adjunctive measures. And several methods were also used to prevent and deal with complications in tumors with different features. In this series the tumor complete necrosis rate (CR)was 95.2% (356/374 tumors). It was higher in 3.5 cm tumors with a CR of 91.3% (156/171 tumors). CR were 95.6% (44/46 tumors) for tumors near the gallbladder, 92.9%(79/85 tumors) for tumors near the diaphragm, 90.9%(40/44 tumors) for tumors near the gastrointestinal tract, 91.2% (31/34 tumors) for tumors near large vessel. In a follow-up period of 2-69 months, the local recurrence rates were 11.7% for HCC and 12.5% for recurrent HCC. The incidence of complications was 2.4% (10/409 sessions), including intraperitoneal hemorrhage (n=2), biliary duct stricture (n=1), hemothorax (n=1), bowel perforation (n=1) and needle tract seeding (n=5). Of these cases, only 3 required operation and the mortality related to RFA was zero in this series. We used Kaplain-Meier method and log-rank test to estimate and compare the survival rate. The 1-, 3-, and 5-year survival rates after RFA were 83.3%, 66.9%, 41.2%, respectively for all HCC patients and 74.6%, 41.3%, 33.6%, respectively for recurrent HCC. Survivals based on TNM stage, Child-Pugh grade, tumor number and tumor size are shown in Table 1. In conclusion, RFA with standard protocol has evolved as a minimally invasive local treatment that could achieve satisfactory outcomes for small liver tumors, and has become an effective and relatively safe alternative for the treatment of advanced tumors and recurrent tumors, which are not suitable for traditional therapy. RFA has broaded the treatment threshold for hepatic malignancies and might become one of the regular treatment methods in focal liver tumor and find wide application. PMID- 16378129 TI - [Skull base meningiomas: a predictive system to know the extent of their surgical resection and patient outcome]. AB - OBJECTIVE: The aim of this study was to build a preoperative predictive system which could provide reliable information about: 1 degrees which skull base meningiomas can be total or partially removed, and 2 degrees their surgical outcome. METHOD: Patient histories and imaging data were reviewed retrospectively from 85 consecutive skull base meningiomas patients who underwent surgery from 1990 and 2002. From the preoperative data, nine variables were selected for conventional statistical analysis as regards their relationship with: 1 degrees total vs partial tumor resection and 2 degrees with patients outcome according to the degree of tumour removal. RESULTS: From the nine variables analysed only two had a statistical association with the type of tumour resection performed (total vs partial) and the patient outcome: 1) arteries encasement and 2) cranial nerves involvement. Upon correlating these two variables with the type of tumour resection performed (total vs partial) and with the Karnofsky'scale to evaluate patients surgical outcome, the following grading groups were identified: Grade I: skull base meningiomas which did not involve cranial nerves or artery or only encased one artery or one cranial nerve. In these cases the incidence of gross tumour resection was 98.3% (p< 0.0001) and the perspective to reach 70 points in the Karnofsky'scale was of 96.5% ( p=0.001). Grade II: skull base meningiomas which involved one cranial nerve and encased, at least, two main cerebral arteries. In these cases, the frequency of total resection, decreased to 83.3% (p<0.0001) and the probability to reach 70 points in the Karnofsky'scale was 70.6% (p=0.001). Grade III: skull base meningiomas which involved two or more cranial nerves and encased several arteries In this group, the frequency of a total resection was of 42.9% (p<0.0001) and the probability of reaching 70 points in the Karnofsky'scale was only 60% (p=0.001). CONCLUSIONS: We propose a preoperative grading system for skull base meningiomas that helps predicting both whether total or partial tumor removal will be achieved during surgery and the immediate postsurgical outcome of the patient. In applying this predictive system we will be able to reduce surgical morbidity, to advance the possibility of a radiosurgical treatment and give a more precise information to the patients and their families about our surgical decision-making process. PMID- 16378130 TI - Analysis of the influence of the variable size on the characteristics and behavior of meningiomas. AB - Seventy-two patients submitted to meningioma surgery at Pedro Hispano Hospital from 1997 to 2001 were reviewed to analyze the association between size (largest diameter of the lesion obtained from imaging examinations) and other variables regarding the biological behavior and clinical outcome of these patients. Statistically significant associations were found between tumor size and location, type of first symptom, type of physical examination, histological grade, surgical complications, postoperative CSF bursae and the need for blood transfusion. Patient's age, gender, duration of first symptom, clinical status at discharge and persistent complaints were not associated to tumor size. There was a trend towards a statistically significant association between tumor size and both grade of resection and persistent deficits. The causes and implications of the findings are discussed. Tumor size is a parameter that may interfere with the neurosurgeon's capacity to treat these patients as well as with their recovery. PMID- 16378131 TI - [Complications of craniofacial resection in anterior skull base tumors]. AB - INTRODUCTION: The development of new radiographic techniques and the refinement of microsurgery and reconstructive surgery have been the responsible of the establishment of craniofacial resection (CFR) as the standard treatment of anterior skull base tumors. Overall complication rates varies from 24-56%, according to a review of recently published series. OBJECTIVES: To describe the complications of CFR in a series of 41 patients and to analyze the management and final outcome. MATERIAL AND METHODS: From 1990 to 2002, 41 patients underwent CFR for tumors involving the anterior cranial base. The extent of the tumor was always assessed with craniofacial CT-scan and MRI. The objective of the surgical treatment was to achieve "on block" removal of the tumor. RESULTS: The average age was 57 years with a male preponderance (63.4%). Squamous cell carcinoma was the most frequent histopathological type of tumor. The tumors were localized in paranasal sinuses in 78% of the cases. Bifrontal craniotomy was performed in 85.4% and unilateral orbitofrontal craniotomy in 14.6% of the cases. The area of facial resection included: ethmoidectomy (60.9%), ethmoido-sphenoidectomy (24.3%), maxillectomy (39%) and orbital exenteration (14.6%). The reconstruction of the floor of the anterior cranial fossa was performed using pedicled pericranial flap (100%), local (34.1%) or microvascular free flaps (21.9%) and split calvarian graft (19.5%). 20 patients (48.7%) developed post-operative complications, CSF leaks (12.1%) and meningitis (7.3%) being the most frequent major complications. The mortality rate was 7.3%. CONCLUSIONS: CFR has become the standard approach for anterior cranial base tumors. Despite its widespread application, the complication's rate ranges between 24-50% and the procedure carries a risk of significant morbidity and even mortality. Improvement of specific aspects of surgical technique and more refined reconstructive methods will decrease the number of complications. PMID- 16378132 TI - [System of dynamic neutralization in the lumbar spine: experience on 94 cases]. AB - OBJECTIVE: To assess the results obtained using the Dynesys system (Dynamic Neutralisation System for the spine), in a group of 94 patients. This new system for treating lumbar degenerative pathologies is based on lumbar stabilisation and preservation of articular function, as opposed to traditional arthrodesis restrictions. MATERIAL AND METHODS: We analyze series of 94 patients in whom this system was used. 62 were males and 32 females with an average age of 46.4 years. The pathologies treated were disc herniation in 27 cases, degenerative discopathy in 54 cases and lumbar channel stenosis in 13 cases. Follow-up was carried out between 14 and 24 months, assessing the clinical picture according to the Oswestry scale and the return to work. RESULTS: The final results on the Oswestry scale were 21.4% with respect to 56.8% prior to the treatment and the return to work was 82%. There was a remission of the sciatica symptoms in almost all the cases, as well as of the lumbar pain, and there was a 60% improvement in the claudication cases. With regard to complications, we would like to point out two cases due to the technique, one because of the wrong positioning of the screws and the other due to the rupture of the pedicle. There were two cases of subcutaneous seroma and two late subclinical infections. CONCLUSIONS: The dynamic neutralisation obtained using this system, should not be considered as an arthrodesis. Treatment using Dynesys enlarges the population of patients candidates for surgery to who initially do not apparently need a standard fixation, but who raise doubt regarding the application of techniques without instrumental support, incorporating the functionality concept as opposed to restricting movement. This system can be defined as a disc prosthesis fitted externally to the disc. We have obtained good results in the majority of our patients, although we believe that the follow-up should be increased. PMID- 16378133 TI - [Related factors with the failed surgery of herniated lumbar disc]. AB - INTRODUCTION: The surgery for herniated disc is the most common operation at the level of the lumbar spine. The failed surgery rates range between 10% and 40%, conforming what is known as Failed Back Surgery Syndrome (FBSS). Return to work after surgery occurs in 70-85% of the cases. There are a few studies analysing the quality of life after the operation. The aims of this study are to know the incidence of the herniated disc lumbar failed surgery in our area, identify those factors influencing its development, and study behavioural parameters as the return to work and the quality of life at a middle term after surgery, also its relation with the success or failure surgery. MATERIAL AND METHODS: A descriptive transversal study of 117 patients operated for herniated lumbar disc during the first six months of the year 2003 is reported. In order to evaluate the incidence of failed surgery and the related factors, the clinical records were retrospectively analyzed. Ninety one patients were interviewed by phone using the Health Questionnaire SF-36, in order to analyze the non clinical factors related to FBSS, such as labour reincorporation, satisfaction with surgery, realization of rehabilitation treatment and quality of life after surgery. For the statistical analysis of the results, we used the program SPSS 11.01. RESULTS: In a sample in which the proportion between both sexes was 1/1, and the middle age was over 45 years [35-54], in which the most frequent clinical symptom was right sciatica, lasting more than 6 months, correlated to disc herniation at L5-S1 level, 37.9% of the patients presented FBSS. Although there were a few patients with reoperation in our study, the incidence of FBSS in these patients was higher (52.9%) than in patients who suffered this surgery for first time (32%). The predictive clinical factors of an unfavourable result in patients operated on for first time were bilateral sciatica, the presence of stenosis associated to herniated disc and comorbidity factors. On the other hand the sociolabor factors identified were a low culture level and those working as drivers, building and service sectors. Return to work occurred in the 64% of the active workers before surgery. Only around 10% of patients were dissatisfied with surgical result and there was a significant relationship between this and the physical function, pain, vitality and emotional status in the SF-36 with the FBSS. CONCLUSIONS: One out of three patients operated of herniated lumbar disc in our area presented failed disc surgery and the return to work occurred in 2 out of three patients active before the operation. The failed surgery patient suffers from pain, that interferes and limits the labour and home activities. Furthermore, the patient presents frequent sensation of fatigue and exhaustion and also emotional problems that contribute to interfere with work and activities of the daily life. PMID- 16378134 TI - Xanthogranuloma of the choroid plexus of the third ventricle: case report and literature review. AB - Xanthogranulomas of the choroids plexus (XG) are benign tumours of debatable etiology which become rarely symptomatic. Only few cases have been studied with MRI. A new case of xanthogranuloma of the third ventricle showing unusual features in the CT and MRI studies in a 47-year-old man with a 2-month history of gait and urinary disturbances and cognitive impairment is reported. The literature concerning clinical and neuroradiological presentation of intracranial xanthogranulomas is reviewed. PMID- 16378135 TI - [Intraventricular meningioma: case report in infancy]. AB - Meningiomas are infrequent tumours in infancy but when they appear in this age group, the intraventricular location is more frequent than in adults. We report the case of a 5-year-old girl with intracranial hypertension and a VI nerve palsy. Computed tomography and magnetic resonance imaging of the brain revealed a tumoral lesion involving the right lateral ventricle with intense enhancement after contrast administration. It was completely resected through a mean temporal circunvolution route. Histological diagnosis was of atypical meningioma. PMID- 16378136 TI - [Internal carotid artery pseudoaneurysm and stenosis: treatment with stents and coils]. AB - PURPOSE: To describe the endovascular management of a patient with post dissection internal carotid pseudoaneurysm and stenosis. CASE REPORT: A 45-year old man presented with left hemyparesia; angiography showed right internal carotid artery occlusion, left carotid stenosis and pseudoaneurysm. A metallic uncovered stent was implanted by means of a femoral approach. Coils were delivered through the uncovered stent and the pseudoaneurysm was excluded. CONCLUSION: The internal carotid percutaneous implantation of a carotid uncovered stent, and embolization through stent in a patient with stenosis and pseudoaneurysm, appears to be a safe procedure without risk of coils migration. PMID- 16378137 TI - [Chronic spontaneous lumbar epidural hematoma]. AB - We report an exceptional case of cauda equina syndrome, caused by a chronic epidural hematoma of the lumbar spine, (L2-L3). A 53 year old man without history of trauma, lumbar punctures or antiagregant medication, suffered from progressive back pain and minor motor deficit in the legs. The etiology, MRI, intraoperative findings and microscopical study are presented and discussed. After surgery there was a complete resolution of the clinical picture. PMID- 16378138 TI - Current pathological and clinical aspects of Buerger's disease in Japan. AB - Since Buerger's disease was first described by Leo Buerger in 1908, many authors have discussed whether it indeed exists and, if so, is a definite clinical entity. Today, Buerger's disease is accepted as a definite vascular disease with a typical clinical picture, natural history, and histopathology; however, the diagnosis of Buerger's disease has been controversial, and the etiology of this vascular occlusive disease has remained unknown because many authors in different countries have their own criteria. Besides, patients with this disease have decreased in number even in Japan, while there has been no change in the number of arteriosclerosis obliterans patients. Currently, only one or two new patients per year are encountered at our outpatient clinic. The purpose of this review is to examine the current epidemiological, pathological, and clinical aspects of Buerger's disease in Japan based on the 222 patient files of our department between 1980 and 2000 and to discuss the pathogenesis, clinical presentation, and various treatment modalities. PMID- 16378139 TI - The nitric oxide donor DETA-NONOate decreases matrix metalloproteinase-9 expression and activity in rat aortic smooth muscle and abdominal aortic explants. AB - Our objective was to examine the role of an exogenous nitric oxide (NO) donor, DETA-NONOate (DETA), on matrix metalloproteinase (MMP)-9, MMP-2, and tissue inhibitor of matrix metalloproteinases (TIMP)-1 expression and activity in interleukin (IL)-1beta-induced rat aortic smooth muscle cells (RA-SMCs) and rat aortic explants (RAEs). RA-SMCs were incubated with IL-1beta (2 ng/ml), an inflammatory cytokine known to induce MMP-9 expression, and increasing concentrations of DETA (0, 1.0, 10, 100 microM; n = 3/group) for 48 hr. RAEs were incubated with IL-1beta (2 ng/mL) and increasing concentrations of DETA (0, 5.0, 50, 100, and 500 microM; n = 3/group) for 48 hr. Media were collected and assayed for NO(x) by the Griess reaction and MMP-9 activity by zymography. Messenger RNA (mRNA) was extracted from cells and analyzed for MMP-9, MMP-2, and TIMP-1 expression levels by quantitative real-time reverse-transcriptase polymerase chain reaction. All statistical analyses were performed by analysis of variance. In RA-SMCs and RAEs, DETA administration resulted in a dose-dependent increase in media NOx concentration (RA-SCM p < 0.01, RAE p < 0.01) and a concurrent decrease in both MMP-9 expression (RASMC p = 0.01, RAE p = 0.01) and activity (RASMC p = 0.04, RAE p = 0.006). There were no significant differences seen in MMP-2 and TIMP-1 expression or activity in response to DETA exposure. DETA decreased IL 1beta-induced MMP-9 expression and activity in both RA-SMCs and RAEs in a dose dependent fashion. In addition, DETA administration had no effect on MMP-2 or TIMP-1 expression or activity in vitro. These data suggest that NO donors may be beneficial in decreasing MMP-9 levels and might serve to inhibit MMP-9-dependent vessel wall remodeling seen during abdominal aortic aneurysm formation. PMID- 16378140 TI - Sequential endovascular repair and pancreaticoduodenectomy for abdominal aortic aneurysm copresenting with periampullary cancer. AB - This report details the case of a patient presenting with a periampullary cancer and an abdominal aortic aneurysm who underwent endovascular repair of the aneurysm prior to pancreaticoduodenectomy. Use of the endovascular approach avoided the need for two major operative interventions. PMID- 16378141 TI - Retrievable inferior vena cava filters: initial clinical results. AB - Anticoagulation is the accepted therapy for patients with thromboembolic disease. When contraindications to anticoagulant therapy are present, however, interruption of the inferior vena cava (IVC) may prevent pulmonary embolism (PE). The objective of this study was to report our early technical and clinical results with retrievable IVC filters (IVCFs) for the prevention of PE. One hundred and twenty-seven multitrauma patients between December 1, 2002, and December 31, 2004, underwent placement of Gunther-Tulip (n = 49), Recovery (n = 41), or OptEase (n = 37) retrievable IVCFs under real-time intravascular ultrasound (IVUS) guidance. All patients had abdominal X-rays to verify filter location. Prior to IVCF retrieval, all patients underwent femoral vein color flow ultrasonography to rule out deep vein thrombosis (DVT) and vena-cavography to assess the IVCF for trapped emboli, filter tilt, or retrained thrombus. Thirty nine patients died of their injuries; no deaths were related to IVCF placement. One PE occurred during follow-up after filter retrieval, and two femoral vein insertion-site DVTs occurred. One hundred twenty (94.4%) of IVCFs were placed without complication at the L2-3 level, as verified by abdominal X-rays. Filter related complications included three groin hematomas (2.9%) and three IVCFs misplaced in the right iliac vein early in our experience (2.3%); these filters were uneventfully retrieved and replaced in the IVC within 24 hr. Sixty-six patients underwent uneventful retrieval of IVCFs after DVT or PE anticoagulation prophylaxis was initiated. Forty-five IVCFs were not removed: 41 due to contraindications due to anticoagulation and four because of trapped thrombus within the filter. The role of retrievable IVCFs continues to evolve, but in this study of 127 patients, prophylactic temporary IVCF placement was simple and safe, prevented fatal PE, and served as an effective "bridge" to anticoagulation. Further investigation of this bedside IVUS technique and the role of temporary IVCFs in different patient populations is warranted. PMID- 16378142 TI - Histological and morphometric study of the arterial route from the intercostal/lumbar artery via the Adamkiewicz artery to the anterior spinal artery using elderly cadavers with or without aortic aneurysms. AB - The Adamkiewicz artery originates from the lower intercostal/lumbar artery, runs along the vertebral roots of the spinal nerve, and communicates with the anterior spinal artery. In graft replacement for an aortic aneurysm, the lower intercostal/lumbar artery should be reconstructed to maintain the arterial blood supply to the lumbar spinal cord via the Adamkiewicz artery. To date, there are few reports on the histological characteristics of the arterial pathway from the aorta to the anterior spinal artery via the Adamkiewicz artery as well as the pathway in the Adamkiewicz artery. In addition, there is a possibility that this arterial pathway was degenerated by the aortic aneurysm. In this study, we attempted to reveal the histological characteristics of the arterial pathway by using elderly cadavers with or without unruptured aortic aneurysm. Specimens were obtained from 190 elderly cadavers (fixed in 10% formalin). These cadavers included 16 cases of aortic aneurysms. We collected fragments of the intercostal/lumbar artery, its dorsal branch, the Adamkiewicz artery, and the anterior spinal artery. After routine histological procedures, several staining procedures were performed for each section. A morphometric study was conducted at the light-microscopic level. The Adamkiewicz artery is composed of a thick intima that contains multiple thick elastic fiber layers and, probably, numerous intimal smooth muscle cells. Among the cadavers, the intimal thickness varied significantly in the intercostal/lumbar artery and the Adamkiewicz artery; it tended to be constant, irrespective of luminar dilatation in the Adamkiewicz artery, in contrast to other arteries. No significant difference was observed between the specimens with or without unruptured aortic aneurysms in all the parameters that were measured. The Adamkiewicz artery appeared to be capable of adapting to acute and chronic changes in systemic status that develop with aging because of its thick intima that contained numerous smooth muscles and elastic fibers. Against various events of systemic circulatory dysfunctions such as hypertension, its unique morphology suggested that this artery may play the critical role of a modulator, or even a barrier, that is intercalated along the arterial route to the lumbar cord. PMID- 16378143 TI - Clinical significance of type II endoleak after endovascular repair of abdominal aortic aneurysm. AB - Type II endoleaks after endovascular repair of abdominal aortic aneurysm (EVAR) are a result of retrograde flow from arterial branches (e.g., lumbar and inferior mesenteric) refilling the aneurysm sac, which has been excluded by the stent graft. Controversy continues with regard to the clinical significance and treatment of type II endoleaks. To develop recommendations for management, we analyzed outcome data from 10 EVAR trials completed over the last 5 years involving a total of 2,617 cases. The incidence of type II endoleak at discharge or 30 days was 6-17%, at 6 months 4.5-8%, and at 1 year 1-5%. Successful resolution of endoleak following secondary interventions was observed in 11-100% of cases. There were 10 conversions to open repair and no ruptures related to type II endoleak. In patients observed for 12 months with computed tomography and/or ultrasound, approximately one-half of type II endoleaks disappeared spontaneously. In the absence of a type I endoleak, our analysis of the current literature suggests that intervention for type II endoleak should be undertaken for abdominal aortic aneurysm sac enlargement occurring after 6 months, persistence for >12 months without abdominal aortic aneurysm sac enlargement, or an aneurysm sac pressure >20% of systolic blood pressure; translumbar aneurysm sac thrombosis and intra-arterial feeding vessel occlusion appear to be prudent management options. PMID- 16378144 TI - Severe chronic venous insufficiency treated by foamed sclerosant. AB - Our objective was to chronicle our experience in using sclerosant foam to treat severe chronic venous insufficiency (CVI). Forty-four patients with 60 limbs severely affected by severe CVI were entered into the study. They had lipodermatosclerosis, CEAP 4 (seven limbs); atrophie blanche or scars of healed venous ulcerations, CEAP 5 (18 limbs); and frank, open venous ulcers, CEAP 6 (35 limbs). Patients and limbs were collected into three groups. In group I, all limbs were treated with compression without intervention. Group II consisted of crossover patients who failed compression treatment. Group III consisted of patients treated promptly with sclerosant foam therapy without a waiting period of compression. A standing Doppler duplex reflux examination was done in all cases. Compression was by Unna boot or long stretch elastic bandaging. Foam was generated from Polidocanol 1%, 2%, or 3% by the two-syringe technique and administered under ultrasound guidance. Posttreatment compression was used for 14 days. In addition to clinical and ultrasound evaluation at 2, 7, 14, and 30 days, venous severity scoring was noted at entry and discharge. In group I, 12 patients were discharged from care within 6 weeks of initiating compression. All eight of the class 6 limbs had healed. Group II consisted of four CEAP class 5 limbs and eight class 6 limbs that had failed to heal with compression. Five of eight venous ulcers healed within 2 weeks, two more healed by 4 weeks, and one required 6 weeks to heal. In group III, 7 of 11 venous ulcers healed within 2 weeks and four more within 4 weeks. Venous severity scores reflected the success of treatment, with the greatest change occurring in group III and the least in group I. Limbs treated with foam had a statistically better outcome than those without (p = 0.041). One patient failed foam sclerotherapy, another had pulmonary emboli 4 months after foam treatment, and a single medial gastrocnemius thrombus was discovered 24 hr after treatment. Treatment of severe CVI with compression and foam sclerotherapy causes more rapid resolution of the venous insufficiency complications and does so without an increase in morbidity. PMID- 16378145 TI - Combined abdominal aortic aneurysm repair and coronary artery bypass: presentation of 13 cases and review of the literature. AB - Coronary artery disease remains the major cause of perioperative mortality after abdominal aortic aneurysm (AAA) repair. The beneficial effect of coronary artery bypass (CAB) before AAA repair in patients with severe coronary artery disease has been proven. The coexistence of a very large or symptomatic AAA and coronary artery disease remains a therapeutic challenge since there is the risk of AAA rupture in the interval between CAB and AAA repair. Combined CAB and aortic aneurysm repair has been suggested for these cases, and results on several series of patients have been published. However, the exact indication for the combined operation remains to be clarified. We present a series of 13 patients who underwent CAB on cardiopulmonary bypass and aortic aneurysm repair as a one-stage procedure. The indication was a large AAA in seven patients and a symptomatic AAA in six patients. In four patients, the aortic reconstruction was performed without the use of cardiopulmonary bypass; in nine patients, the aortic reconstruction was performed under partial cardiopulmonary bypass. Thirty-day mortality was 15%. Major morbidity was 31%. All major complications were due to excessive bleeding and occurred in patients who had AAA repair performed with partial cardiopulmonary bypass, suggesting that prolonged bypass time represents a major source of morbidity. A detailed review of the literature is presented. From the evidence available we suggest that the combined procedure can be recommended only for patients with very high rupture risk, such as in symptomatic AAA. In all other cases, the staged approach--CAB followed by AAA repair 2-4 weeks later--is preferable. During the combined procedure, cardiopulmonary bypass support during AAA repair should be used only in patients with clear evidence of hemodynamic instability. PMID- 16378146 TI - Vascular closure stapler clip anastomosis decreases aortic cross-clamping time compared to interrupted nonabsorbable and running absorbable sutures in growing pigs. AB - The latest generation in titanium clip application systems, the vascular closure staples (VCS) system avoids intimal lesions and therefore minimizes the subsequent hyperplastic reaction, while at the same time enhancing distensibility and vascular growth. Moreover, VCS clips allow the surgeon to perform vascular anastomosis easily and faster than conventional suture. This system may become the option of choice for vascular reconstruction in pediatric surgery where, as in the case of aortic and transplant surgery, decreasing vascular occlusion times may influence the outcomes. The aim of this study was to determine whether VCS metallic clips would allow shorter anastomosis times than conventional interrupted polypropylene or running Dexon suturing in end-to-end anastomosis performed in the abdominal aorta of young pigs. Twenty-four domestic swine, 45 days old, were used for this study. All animals were subjected to an end-to-end anastomosis in the abdominal aorta using VCS clips, interrupted polypropylene, or running Dexon suture. Aortic cross-clamping time was significantly shorter in the VCS clips group (4.02 +/- 0.72 min), compared to interrupted polypropylene (21.89 +/- 1.93 min) or running Dexon (9.82 +/- 3.97 min) suture. VCS clips are easy to use, and therefore may aid surgeons to significantly decrease the time needed for performing an end-to-end anastomosis in the abdominal aorta, thus decreasing cross-clamping time when compared to interrupted polypropylene or running Dexon sutures. PMID- 16378147 TI - Quality of life after surgery for varicose veins and the impact of preoperative duplex: results based on a randomized trial. AB - In a prospective randomized study, we found that the addition of a preoperative duplex scan before varicose vein (VV) surgery reduced recurrences and reoperations after 2 years. The aim of the present study was to investigate whether this correlates with an improved quality of life (QoL). We studied 293 patients scheduled for VV surgery with or without preoperative duplex. QoL was assessed preoperatively at 1 month, 1 year, and 2 years with the Short Form-36 (SF-36). Scores were compared with matched reference groups from the Swedish population. The 237 complete responders (81%) had a mean age of 47 (range 22-73) years, 169 (71%) were women, and 43 (18%) had skin changes. Both groups of VV patients scored significantly worse than the reference group in the domain Bodily Pain preoperatively (p < 0.001) and better after 1 year (p = 0.04), with no difference found after 2 years. There was no significant difference in QoL between the duplex and control groups at any time. We conclude that preoperative duplex before VV surgery did not significantly improve QoL after 2 years in spite of improved surgical results. VV surgery per se improved QoL as measured with the SF-36. PMID- 16378148 TI - Ascorbic acid (vitamin C) and iloprost attenuate the lung injury caused by ischemia/reperfusion of the lower extremities of rats. AB - The objectives of this study were to compare the protective effects of ascorbic acid and iloprost on lung injury caused by ischemia reperfusion (I/R) of the lower extremities of rats. Wistar albino rats (n = 34) were divided into five groups. In the I/R group (n = 6), the aorta was cross-clamped for 3 hr, followed by 1 hr of reperfusion. In the vitamin C group (n = 8), animals were pretreated with 100 mg/kg ascorbic acid via the left jugular vein before aortic cross clamping. In the iloprost group (n = 8), animals were pretreated with 20 ng/(kg x min) iloprost by constant intravenous infusion via the left jugular venous cannula. In the sham group (n = 6), the abdomen was left open at the same period and a juguler venous line was established. In the control group (n = 6), lungs were removed and blood samples taken immediately after sternotomy. No treatment was given in this group. After both lungs were removed, biochemical parameters were measured and histopathological evaluation was made. Although the arterial blood pO2 and HCO3 levels were statistically significantly high in both the vitamin C and iloprost groups compared to the I/R group, plasma malondialdehyde (MDA) levels were significantly low. Meanwhile, the MDA levels in the lung tissue were significantly low in the vitamin C group compared to the I/R group. The MDA level in the lung tissue in the iloprost group was also low compared to the I/R group, but it was not statistically significant. The lungs of the I/R group displayed intense interstitial leukocytic infiltration in histopathological examination compared to the other groups. Pretreatment of animals with iloprost and vitamin C significantly decreased the pulmonary injury characterized by decreased plasma leukocyte sequestration. The results suggest that both vitamin C and iloprost are useful agents for attenuating the lung injury caused by increased oxidative stress and neutrophil accumulation after a period of I/R of the lower extremities. PMID- 16378149 TI - Utility of surgery for ruptured abdominal aortic aneurysm. AB - Our aim was to assess the utility of surgery for ruptured abdominal aortic aneurysm (RAAA) using the number of quality-adjusted life years (QALYs) in a retrospective study with cross-sectional quality-of-life (QoL) evaluation. During a 7-year period up to 2002, 242 of 269 (90%) patients with RAAA underwent surgery. Survivors were sent the EQ-5D self-administered questionnaire to assess their long-term outcome. EQ-5D single index values were calculated for each survivor and combined with age- and sex-adjusted Finnish life tables to obtain QALY estimates. Total hospital mortality (90-day) and operative mortality (30 day) were 140 of 269 (52.0%) and 106 of 242 (43.8%), respectively. Of the 129 surviving patients, 111 were available for QoL evaluation. The response rate was 85%. The mean (range) number of QALYs after RAAA was 4.1 (0-30.9) for all and 8.5 (0.2-30.9) for hospital survivors. Young age and low Glasgow Aneurysm Score were associated with a high number of QALYs irrespective of the statistical method used for analysis. Successful repair of RAAA was able to lend considerable benefit as measured by QALYs. PMID- 16378150 TI - Repair of anastomotic abdominal aortic pseudoaneurysm utilizing sequential AneuRx aortic cuffs in an overlapping configuration. AB - Anastomotic aortic pseudoaneurysm is a known late complication following aortic repair and presents a considerable surgical challenge. We herein evaluate the endovascular alternative of using sequential AneuRx aortic cuffs to bridge the degenerative anastomotic pseudoaneurysms as a definitive treatment. Over a 3-year period, six patients with a mean age of 68.7 years (range 58-75) were identified who had proximal anastomotic aortic pseudoaneurysms secondary to previously implanted bifurcated aortic grafts (mean 15, range 12-20 years) following open surgical correction of aortoiliac occlusive disease. Five patients (83%) presented with concomitant palpable femoral anastomotic pseudoaneurysms and one patient (16%) had a pulsatile abdominal mass. All patients had computed tomographic (CT) scans confirming proximal anastomotic pseudoaneuryms without evidence of infection. The mean diameter of the pseudoaneurysms was 5.3 cm (range 4.0-7.0). Five patients were treated with endovascular methods, while one patient was not suitable for endovascular repair due to the diameter of the native aorta as seen on imaging study at the time of the procedure. AneuRx aortic extender cuffs (3.75 cm length) were deployed sequentially in five patients via a femoral approach. Devices were overlapped approximately 1.5 cm in order to achieve total exclusion of the pseudoaneurysms, and all concomitant femoral aneurysms were repaired surgically at the same time. Successful exclusion of the anastomotic pseudoaneurysm was achieved in four patients (80%) using a combination of two or three overlapping aortic cuffs. One patient had a small residual endoleak that had sealed by 1 month, evidenced by follow-up CT. The renal arteries were preserved in all patients. The average estimated blood loss and operative time were 355 +/- 25 cc and 84 +/- 21 min, respectively. The average length of hospital stay was 2.1 days, and there was no mortality or major morbidity. All patients underwent CT scanning surveillance at 6 and 12 months and yearly afterward. There was no evidence of late endoleak, aneurysmal expansion, or device migration during the mean follow-up of 10 months, ranging 6-27 months. Our study demonstrated that utilizing sequential AneuRx aortic cuffs applied in an overlapping configuration is an effective strategy for degenerative aortic anastomotic pseudoaneurysm from previously placed aortic grafts. Additionally, our study suggests this unique endovascular technique is an ideal alternative for creating a customized tube graft in challenging cases, particularly in high-risk patients. PMID- 16378151 TI - Highlights of the recent literature on abdominal aortic aneurysm research. PMID- 16378152 TI - Effects on skeletal muscle glutathione status of ischemia and reperfusion following abdominal aortic aneurysm surgery. AB - Glutathione (GSH) is an important endogenous scavenger against reactive oxygen species. Elective abdominal surgery without ischemia and reperfusion leads to decreased muscle GSH concentrations 4-72 hr postoperatively without altering GSH redox status. In the present study, we investigated to what extent muscle GSH status was affected during and following elective abdominal aortic aneurysm repair. From patients (n = 10) undergoing abdominal aortic repair, thigh muscle specimens were taken preoperatively, at maximal ischemia, and at 10 min and 4, 24, and 48 hr of reperfusion. Specimens were analyzed for GSH, amino acids, and energy-rich compounds. At maximal ischemia, phosphocreatine decreased by 37% (p < 0.05) and lactate and creatine increased by 274% and 57% (p < 0.001 and 0.05), respectively, indicating ischemia during the clamping of aorta. Adenosine triphosphate, on the other hand, remained unaltered during the entire study period. Total GSH (tGSH) decreased by 46% at 24 hr and by 43% at 48 hr of reperfusion (p < 0.001), while reduced GSH decreased by 48% at 24 hr and by 44% at 48 hr (p < 0.001). The redox status (GSH/tGSH) of GSH and oxidized GSH remained unaltered. Among the constituent amino acids of GSH, glycine and cysteine remained unaltered while glutamine and glutamate decreased by 55% and 55%, respectively (p < 0.001). Abdominal aortic aneurysm repair induces metabolic alterations characteristic for ischemia. The antioxidative capacity in terms of muscle levels of GSH was decreased. However, the oxidative stress during reperfusion did not change GSH status more than what has been reported following abdominal surgery without ischemia and reperfusion. The results indicate that the oxidative stress elicited by elective abdominal aortic aneurysm repair is outbalanced by a compensated GSH metabolism not giving rise to an increased amount of oxidized GSH or an altered GSH redox status. PMID- 16378153 TI - Going beyond the kidney disease outcomes quality initiative: hemodialysis access experience at Kaiser Permanente Southern California. AB - In 1996, as part of Kaiser Permanente Southern California's participation in the Medicare End-Stage Renal Disease Managed Care Demonstration Project, a multidisciplinary continuous quality improvement (CQI) committee was formed, which included nephrologists, vascular surgeons, interventional radiologists, care managers, a renal quality-improvement nursing director, and renal program administrators. The goal of this report was to analyze the impact of this CQI program on hemodialysis outcomes within the organization. Kaiser Permanente is a national, integrated, nonprofit, staff model health maintenance organization with 8 million members. The southern California region has 3.1 million members and currently manages the health care of 3,700 hemodialysis patients, 300 peritoneal dialysis patients, and 1,000 kidney transplant patients. Thirty-one vascular surgeons and 29 interventional radiologists provide for their hemodialysis access needs. The Kidney Disease Dialysis Outcomes Quality Initiative (K/DOQI) guidelines were adopted, as well as measures to perform more venous transpositions and less common arteriovenous fistulas (AVFs) before graft placement. The outcomes assessed included incidence and prevalence of AVFs, grafts, and catheters; replacement access with AVFs; and combined AVF and graft thrombosis episodes per patient per year. Primary AVF incidence rates increased from 27% in 1997 to 88% in 2003. AVF prevalence rates increased from 30% in 1997 to 62% in 2003. Replacement access which is an AVF increased from 26% in 1998 to 58% in 2003. Yearly thrombosis episodes/patient decreased from 0.62 in 1998 to 0.34 in 2003. Catheter usage as of 2003 comprised an incidence of 65% and prevalence (> or =90 days) of 13%, which was essentially unchanged from 1999, despite improvements in fistula usage and thrombosis rate. The rate of AVF prevalence can be increased dramatically, exceeding the 40% K/DOQI recommendation, by using the CQI process. Increased prevalence of AVF is associated with a lower yearly incidence of thrombosis episodes/patient. Reducing excessive catheter usage appears to be a more difficult problem. PMID- 16378154 TI - Aortoiliac endarterectomy: a lost art? AB - Endarterectomy was first performed on a superficial femoral artery in 1946 by Cid dos Santos and subsequently on the abdominal aorta by Wylie in 1951. During the 1950s and 1960s, aortoiliac endarterectomy (AIE) was the standard procedure for treatment of aortoiliac occlusive disease. When prosthetic graft material became available, aortobifemoral bypass (ABFB) replaced AIE in most cases because occlusive disease commonly affects the external iliac arteries also, which were difficult to endarterectomize. As a result, aorto-common iliac endarterectomy became almost a lost art. However, we believe there is still a place for AIE in selected patients based on a review of our results with the procedure. We reviewed 205 patients who survived 10 years after undergoing operation for aortoiliac occlusive disease by either aorto-common iliac endarterectomy (n = 39) or ABFB (n = 166). Ten-year primary patency was 89.2% for AIE and 78% for ABFB. Graft infection or aneurysmal formation occurred in 5% of ABFB and 0% of AIE cases. Ten male patients who underwent AIE for leg and hip claudication with positive penile/brachial indices of 70%) carotid stenosis had 12 asymptomatic intracranial aneurysms (IAs). None had postoperative stroke or died, and none had surgical or endovascular IA treatment before or after CEA. In an average 5-year follow-up, no patients had subarachnoid hemorrhage. Having an IA does not seem to be an additional risk factor for CEA, nor does CEA seem to increase the chance of IA rupture. PMID- 16378156 TI - Clinicopathologic features associated with having four or more metastatic axillary nodes in breast cancer patients with a positive sentinel lymph node. AB - BACKGROUND: The survival benefit of a completion axillary lymph node dissection (ALND) in patients after removal of a metastatic sentinel lymph node (SLN) is uncertain and is under study in ongoing clinical trials. The completion ALND remains necessary, however, for the identification of cases with at least four metastatic lymph nodes, in which extended-field locoregional and/or postmastectomy radiation will be recommended. Our goal was evaluate clinicopathologic features that might serve as surrogates for determining which patients with a positive SLN are likely or unlikely to belong to this high-risk subset. METHODS: Records were reviewed for 285 patients from 2 comprehensive cancer centers who underwent completion ALND after resection of a metastatic SLN from 1995 to 2002. Clinicopathologic features were analyzed by univariate and multivariate logistic regression. Forty-one cases (14%) were found to have at least four positive nodes after ALND. RESULTS: Fisher's exact test revealed the following features to be significantly (P < .05) associated with having four or more nodal metastases: tumor size >2 cm, lymphovascular invasion, an increasing ratio of positive SLNs to the total number of resected SLNs, extranodal extension, and the size of the SLN metastasis. Patients whose largest SLN metastasis was <2 mm had only a 1.4% risk of having four or more metastatic nodes (P < .0001). CONCLUSIONS: We conclude that patients with SLN micrometastases face an extremely low likelihood of having extensive nodal disease on completion ALND. Patients with larger primary tumors, lymphovascular invasion, and extranodal extension are more likely to have ALND findings that will affect their cancer management. PMID- 16378157 TI - Preoperative profiling of symptomatic breast cancer by diagnostic core biopsy. AB - BACKGROUND: Precise preoperative profiling of breast tumors could facilitate fuller consideration of (neo)adjuvant therapies. METHODS: Diagnostic core biopsy (DCB) accuracy in profiling the primary tumor was prospectively studied in 95 patients with operable breast cancer. The histological type and grade (hematoxylin and eosin staining) and membrane receptor status (semiquantitative immunohistochemistry for estrogen [ER] and progesterone [PR] receptors, as well as Her-2 antigen expression) were assigned by the DCB before surgery. These measures were then compared with those of the definitive surgical specimen available after operation. RESULTS: DCB correctly ascribed tumor type and grade and ER, PR, and Her-2 receptor status in most cases (correlating exactly in 97.5%, 77%, 68%, 71%, and 60%, respectively) with at least moderate concordance (weighted kappa, >.41). When miscategorized, DCB consistently tended to upscore the receptor stain intensity compared with the surgical specimen (22%, 19%, and 27% had higher ER, PR, and Her-2 categorical scores, respectively). ER H-scores correlated best in specimens that stained strongly (224.4 +/- 3 vs. 215.5 +/- 5) and were significantly higher on DCB in those that stained either moderately (195.6 +/- 8.2 vs. 156.8 +/- 5.1; P < .0001) or weakly (157.1 +/- 24.8 vs. 81.4 +/- 4; P = .02). DCB accurately identified all tumors with clinically important ER and Her-2 expression. Furthermore, it promoted three patients into the therapeutically significant range of ER (n = 1) or Her-2 (n = 2) expression. ER negativity on DCB (n = 25) indicated a high-grade tumor (88%), although 11 (44%) patients also overexpressed Her-2. Significant Her-2 expression (n = 16) on DCB predicted the tumor as being poorly differentiated (80%) and both ER and PR negative (67%). CONCLUSIONS: DCB accurately profiles clinically relevant measures of primary tumor cell differentiation. It also reliably categorizes patients with regard to (neo)adjuvant therapy before radical surgery is attempted. PMID- 16378158 TI - Primary hyperparathyroidism in multiple endocrine neoplasia type 1: individualized management with low recurrence rates. AB - BACKGROUND: To evaluate the outcomes in different surgical modalities for primary hyperparathyroidism in multiple endocrine neoplasia type 1 (MEN1) patients, intraoperative findings from a single surgeon were studied to investigate a potentially improved modality of parathyroidectomy (PTx). METHODS: All 22 patients had PTx by a single surgeon in the past 21 years. Three modalities of PTx were used, depending on the operative findings, after all parathyroids and the thymus were identified. If fewer than three glands were enlarged, selective removal of the enlarged glands with or without biopsy of a normal-appearing gland was performed (selective PTx); if all glands were enlarged, either a subtotal PTx leaving a 50-mg remnant in situ or a total PTx with autotransplantation (TPTx + AT) was performed. RESULTS: There were 7 men and 15 women, aged 22 to 67 years (average, 43 years). Sixteen had familial and six had sporadic MEN1. They underwent 23 operations, including 11 selective PTx, 6 subtotal PTx, and 6 TPTx + AT. On follow-up for 1 to 19 years, only one patient (4.6%) had recurrent hyperparathyroidism 5.5 years after subtotal PTx. Others had either normocalcemia (n = 14; 63.6%) or hypocalcemia (n = 7; 31.8%). Those who had either a subtotal PTx or TPTx + AT had a significantly higher rate of postoperative hypocalcemia than those who had a selective PTx (9.9% vs. 54.5%; P = .032; Fisher's exact test). CONCLUSIONS: Primary hyperparathyroidism in our MEN1 patients was less aggressive than that reported in the literature. Selective PTx according to the intraoperative findings achieved optimal outcomes. PMID- 16378159 TI - Interferon gamma-inducible protein 10 selectively inhibits proliferation and induces apoptosis in endothelial cells. AB - BACKGROUND: Interferon gamma-inducible protein 10 (IP-10) has antitumor effects in various murine models. The IP-10 receptor has two distinct splice variants, CXCR3A and CXCR3B, that have paradoxical effects after ligand-receptor interaction. METHODS: To characterize the putative antiangiogenic effects of IP 10, we measured proliferation rates and apoptosis in human umbilical vein endothelial cells (HUVECs), fibroblasts, and A375 melanoma or WIDR adenocarcinoma cell lines after exposure to the recombinant protein. CXCR3A (activating) and CXCR3B (inhibitory/proapoptotic) messenger RNA (mRNA) expression levels in fibroblasts, 2 human tumor cell lines, T lymphocytes, and HUVECs of varying cell densities were characterized. RESULTS: IP-10 resulted in dose-dependent and selective inhibition of proliferation and countered the proliferative effects of vascular endothelial growth factor in HUVECs but did not affect fibroblasts or 2 human tumor cell lines. In addition, IP-10 resulted in potent and selective induction of apoptosis in HUVECS but had no effect on fibroblasts or A375 melanoma. Confluent HUVECs had a predominance of mRNA for the CXCR3B splice variant by reverse transcriptase-polymerase chain reaction, and the ratio of CXCR3B to CXCR3A mRNA was >40 in HUVECs, compared with or=30 kg/m2) was the only independent predictor of the presence of an ANA titer of 1 : 80 or more (P = 0.026). CONCLUSIONS: The AIH score without liver biopsy findings was not useful for diagnosing AIH in NAFLD patients. In patients with elevated ANA titers and risk factors for NAFLD, it is very important to perform a liver biopsy to make a definitive diagnosis before treatment. PMID- 16378178 TI - Differential diagnosis of nodular lesions in cirrhotic liver by post-vascular phase contrast-enhanced US with Levovist: comparison with superparamagnetic iron oxide magnetic resonance images. AB - BACKGROUND: We investigated the diagnostic utility of post-vascular phase contrast-enhanced ultrasonography (US) and superparamagnetic iron oxide (SPIO) enhanced magnetic resonance imaging (MRI) as compared to the histological diagnosis of differential grades of hepatocellular carcinomas (HCCs). METHODS: Forty-nine patients with histologically characterized liver nodules (well differentiated HCC, n = 20; moderately differentiated HCC, n = 19; poorly differentiated HCC, n = 1; dysplastic nodule, n = 9) received contrast-enhanced US and SPIO-MRI. Subsequently, we quantitatively evaluated the relationships between the images of the nodules and their histological diagnosis and differential grades. RESULTS: The ratio of the echogenicity of the tumorous area to that of the nontumorous area with post-vascular phase contrast-enhanced US (post-vascular phase ratio) decreased as nodules became less differentiated (P < 0.05; Kruskal-Wallis test). The ratio of the intensity of the nontumorous area to that of the tumorous area on SPIO-enhanced MR images (SPIO intensity index) also decreased as nodules became less differentiated (P < 0.01). The post-vascular phase ratio correlated with the SPIO intensity index for HCCs and dysplastic nodules (r = 0.76). The conformity of the result from the post-vascular phase contrast-enhanced US and SPIO-MRI was 96%. CONCLUSIONS: Contrast-enhanced US is a valuable method for predicting the histological grade of HCCs in cirrhotic patients, and may be a good alternative to SPIO-enhanced MRI. PMID- 16378179 TI - Diagnosing clinical subsets of autoimmune liver diseases based on a multivariable model. AB - BACKGROUND: Diagnosing autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC), primary sclerosing cholangitis, and other autoimmune liver diseases remains an imperfect process. We need a more accurate, evidence-based diagnostic system. METHODS: We conducted a national survey and identified 988 cases of liver disease which did not satisfy the inclusion criteria for any liver disease of known etiology. We expected these cases to include autoimmune liver disease (AILD) and its variant forms. We selected 269 prototype cases for which histological re-evaluation of liver biopsy by independent expert hepatopathologists and the original diagnosis coincided. We did a multiple logistic regression analysis to determine explanatory variables that would distinguish cases of AIH and PBC from those of non-AIH and non-PBC, respectively. We constructed a multivariable diagnostic formula that gave AIH and PBC disease probabilities and validated it in a study of an additional 371 cases (validation group). RESULTS: Based on the results of the statistical analysis, we selected three laboratory tests and four histological features as independent variables correlated to the diagnosis of both AIH and PBC. For the validation group, assuming that the original diagnosis was correct, the sensitivity and specificity for AIH were 86.3% and 92.4%, respectively. For PBC the sensitivity and specificity were 82.5% and 63.7%, respectively. A detailed analysis of inconsistent cases showed that the diagnosis based on the formula had given the correct diagnosis, for either AIH or PBC, except for 5 cases (1.3%) in which disease probability was low for both. CONCLUSIONS: A seven-variable formula based on three laboratory tests and four histological features gives significant information for the diagnosis of AILD. PMID- 16378180 TI - Crohn's disease with life-threatening hemorrhage from terminal ileum: successful control by superselective arterial embolization. AB - A case of life-threatening lower gastrointestinal hemorrhage from Crohn's disease is reported. Several promising studies have recently been published that describe superselective embolization for the treatment of massive lower gastrointestinal hemorrhage in patients with bleeding colonic diverticular disease and angiodysplasia, and success rates of 74%-93% have been reported. But in patients with Crohn's disease, successful superselective embolization has rarely been reported. This is a report of successful superselective embolization in a patient with Crohn's disease; this should be the initial treatment of choice in Crohn's disease in an attempt to avoid surgical resection, because repeated resections predispose patients to the development of short-bowel syndrome. PMID- 16378181 TI - Granulocyte colony-stimulating factor-producing combined hepatocellular/cholangiocellular carcinoma with sarcomatous change. PMID- 16378182 TI - Diagnostic challenges from inadvertent retrieval of adrenal gland tissue with transjugular liver biopsy. PMID- 16378183 TI - Brain or gut? Site of action of adrenomedullin to regulate gut motility. PMID- 16378184 TI - Is the new diagnostic scoring system of Zeniya and colleagues useful for autoimmune liver diseases? PMID- 16378185 TI - Common fragile genes and digestive tract cancers. AB - FRA3B and FRA16D are the most sensitive common chromosomal fragile site loci in the human genome. Two tumor suppressor genes, the fragile histidine triad (FHIT) gene and the WW domain-containing oxidoreductase (WWOX), map to the common fragile sites, FRA3B and FRA16D, respectively. Interestingly, FHIT and WWOX have similarities: for example, they are both larger than 1 Mb and encompass fragile sites, they both show frequent allelic loss regions in various human cancers, they both span a region of homozygous deletion in multiple cancers, and they both frequently show aberrant transcripts. The development of human cancers, including digestive tract cancers, is strongly associated with exposure to environmental carcinogens. Common fragile sites are very sensitive to this type of exposure, and the resulting DNA damage leads to the inactivation of genes such as FHIT and WWOX. We present an overview of these two common fragile genes, namely FHIT/FRA3B and WWOX/FRA16D, in digestive tract cancers. PMID- 16378186 TI - Arterio-jugular differences in serum S-100beta proteins in patients receiving selective cerebral perfusion. AB - PURPOSE: The early increase in serum S100beta after cardiopulmonary bypass (CPB) seems to be derived from an extracerebral source. To exclude contamination, we investigated the arterio-jugular differences in S100beta levels in patients receiving selective cerebral perfusion (SCP). We also evaluated the brain protective effect of SCP by comparing the arterial S100beta levels with those in patients undergoing coronary artery bypass grafting (CABG). METHODS: We measured arterial and jugular venous levels of S100beta in ten patients undergoing aortic arch repair with SCP for up to 12 h postoperatively (SCP group). We also measured arterial levels of S100beta in nine patients undergoing CABG (CPB group). RESULTS: There was no incidence of hospital death or stroke. The arterial levels of S100beta in both groups were comparable and peaked just after the conclusion of CPB. The arterial and jugular venous levels of S100beta were almost equivalent. The arterio-jugular differences in S100beta levels were negligible, even in our SCP-group patient with postoperative delirium, who had a peak value three times higher than the other patients. CONCLUSIONS: The arterio-jugular differences in S100beta did not clarify the origin of their increase. Thus, measuring the jugular venous levels of S100beta in patients without postoperative clinical neurological deterioration would be of little benefit. However, SCP seems to protect the brain against S100beta release as effectively as conventional CPB. PMID- 16378187 TI - Minimal extrathyroid extension does not affect the relapse-free survival of patients with papillary thyroid carcinoma measuring 4 cm or less over the age of 45 years. AB - PURPOSE: In the most recent version of the UICC TNM classification system for thyroid carcinoma, tumors with minimal extrathyroid extension were classified as T3. In this study, we investigated whether this upgrading is appropriate for papillary thyroid carcinoma. METHODS: We investigated the difference in the relapse-free survival (RFS) rate between patients with tumors having no, minimal, and massive extrathyroid extension in a series of 502 patients over the age of 45 years. RESULTS: Patients with tumors showing massive extension showed a worse RFS rate except for those with tumors measuring 1 cm or less. However, there was no significant difference in RFS between tumors measuring 4 cm or less showing no or minimal extension. In an investigation of 409 patients without any clinically apparent node metastasis, the RFS of patients with tumors larger than 4 cm with massive extension was significantly worse than those with tumors measuring 4 cm or less, while the RFS of patients with tumors with either no or minimal extension did not depend on the tumor size. CONCLUSIONS: These findings suggest that tumors with minimal extension should be classified to have the same T grade as those without such extension, and upstaging of such tumors is therefore not appropriate. PMID- 16378188 TI - A new method of thermo-chemotherapy using a stent for patients with esophageal cancer. AB - PURPOSE: Many patients with advanced esophageal cancer have a stent inserted in the esophagus, but very little else can be done. We devised a new method of delivering thermotherapy using an implant heating system (IHS) with a metallic stent. To our knowledge, there are no other reports of thermotherapy using metallic stents. We report our clinical results of treating patients with this new technique. METHODS: A metallic stent was placed in the esophagus and heated for thermotherapy. This was combined with simultaneous chemotherapy in 13 patients and radio-chemotherapy in 5 patients. Each thermotherapy session consisted of heating the stent to 50 degrees C for 10 min. These 18 patients received a collective 52 sessions of thermotherapy. The tumor was excised after this treatment in 5 of these 18 patients. RESULTS: We evaluated 17 of the 18 patients, after the exclusion of 1 patient who underwent radio-chemotherapy before placement of the stent. There was 1 complete response (CR) and 12 partial responses (PR), accounting for 76%. The patient with a CR had no residual cancer cells detected by pathologic examination after surgical resection. Thermotherapy proved effective in 8 (89%) of 9 patients who received this treatment at least three times. It was effective in all (100%) patients who underwent concomitant radio-chemotherapy. CONCLUSIONS: Our results show that thermotherapy using a stent can improve the effectiveness of combination therapy, suppress local tumor growth, and enhance quality of life over a long period. PMID- 16378189 TI - Novel clinical staging for patients with end-stage gastrointestinal carcinoma. AB - PURPOSE: We created a new clinical staging system for end-stage gastrointestinal (GI) carcinoma to clarify the therapeutic goals for these patients. METHODS: Data were obtained from a retrospective review of medical charts. Based on daily clinical observation of 144 patients with end-stage GI carcinoma, we classified the terminal stages as A, B, C, and D. RESULTS: The mean durations of terminal stages A, B, C, and D were 19, 16.6, 6.6, and 1.8 days, respectively, in patients with end-stage gastric cancer and 28.5, 9.1, 5.4, and 1.9 days, respectively, in patients with colorectal cancer. Moreover, 88.0% of patients with gastric carcinoma and 82.6% of patients with colorectal carcinoma passed through terminal stages A, B, C, and D sequentially. The patients in terminal stage B experienced temporary relief of symptoms, but those in terminal stage C did not (P < 0.05). CONCLUSIONS: These terminal stages can easily be judged by clinical observation and may be an effective new tool with which to manage patients with end-stage GI carcinoma and their families. PMID- 16378190 TI - Modified double-stapling technique in low anterior resection for lower rectal carcinoma. AB - PURPOSE: The original double-stapling technique (DST) using a standard linear stapler horizontally can be difficult in patients with a narrow pelvis or an ultralow anastomosis. We review our experience of performing a modified DST (IO DST) with vertical division of the rectum achieved using an endostapler. METHODS: We retrospectively studied the clinical outcomes of 90 patients who underwent low anterior resection (LAR) for lower rectal carcinoma. Low anterior resection was performed with IO-DST in 34 patients (IO-DST group), with the single-stapling technique (SST) in 47 (SST group), and with per anal anastomosis (PAA) in 9 (PAA group). RESULTS: The distances from the anal verge to the tumor and to the anastomosis were significantly shorter in the IO-DST group than in the SST group (5.8 cm, 4.0 cm vs 7.0 cm, 5.0 cm, respectively), whereas it was equivalent in the IO-DST and PAA groups (5.0 cm, 4.0 cm). Blood loss was less in the IO-DST group than in the SST and PAA groups (400 ml vs 578 ml and 950 ml, respectively). The operative time was shorter in the IO-DST group than in the PAA group (281 min vs 327 min, respectively). There were no significant differences in the length of the distal surgical margin among the three groups. The IO-DST group patients suffered less bowel frequency than the SST group patients 1 month after surgery (2.5 times/day vs 4.0 times/day, respectively) and less than the PAA group patients more than 1 year after surgery (2.0 times/day vs 3.5 times/day, respectively). There were no significant differences in the incidence of complications or local recurrence among the three groups. CONCLUSIONS: IO-DST is a feasible and safe procedure for performing low anastomosis, which results in less bowel frequency after LAR for lower rectal carcinoma. PMID- 16378191 TI - Implementation of a scoring system for assessing difficult cholecystectomies in a single center. AB - PURPOSE: Laparoscopic cholecystectomy (LC) is the treatment of choice for symptomatic cholelithiasis, but sometimes conversion to open cholecystectomy (OC) is necessary. The difficulty of LC or the risk of conversion to OC can be predicted by assessing some preoperative variables. We evaluated the efficacy of the "risk score for conversion from laparoscopic to open cholecystectomy" (RSCLO), which was recently developed by Kama et al. (Am J Surg 2001; 181:520), in a single center. METHODS: We recorded the RSCLO values of 571 patients admitted to undergo LC in the Department of General Surgery, Firat University Hospital, between June 2001 and June 2004. RESULTS: The mean RSCLO score of 19 patients who needed conversion to OC was significantly higher than that of the patients who underwent successful LC, at 16.2 (range, -9 to 41) vs -5.7 (range, 20 to 25) (P < 0.001). The RSCLO was well correlated with conversion to OC. The sensitivity and specificity rates for RSCLO determining the risk of conversion to OC were 100% and 96%, respectively, and its positive and negative predictive values were 43% and 100%, respectively. CONCLUSION: We think that RSCLO could be used to define the term "difficult LC" more accurately and assist in selecting the most appropriate operation. PMID- 16378192 TI - Tamoxifen and the risk of endometrial cancer in Japanese women with breast cancer. AB - PURPOSE: There is ongoing debate about whether tamoxifen for breast cancer is associated with a risk of endometrial cancer in Japanese women. We conducted a study to investigate this further. METHODS: We conducted a retrospective hospital based cohort study. A total of 674 consecutive patients underwent surgery for primary breast cancer between January 1989 and December 1998. By December 2003, endometrial cancers had been diagnosed in six of these patients. Based on medical records, we evaluated the potential risk factors for endometrial cancer, including age, menopausal status, obesity, parity, diabetes mellitus, hypertension, and tamoxifen. The 674 patients were divided into three groups based on the cumulative duration of tamoxifen use (A, <2 years vs B, 2-5 years vs C, >5 years). To examine the relationships between endometrial cancer and tamoxifen (and other factors), the hazards ratio (HR), 95% confidence interval (CI), and two-sided P value for endometrial cancer associated with each variable were calculated by the Cox regression method. RESULTS: Endometrial cancer was found in 1/318 (0.31%) patients in group A, 3/247 in group B, and 2/109 in group C. In a multivariate analysis no variable was significant, but tamoxifen use for longer than 5 years (group C) was closely correlated with endometrial cancer (HR = 7.92, CI = 0.69-90.89, P = 0.096). CONCLUSION: Although our data did not reach significance, they support a link between long-term tamoxifen and the development of endometrial cancer in Japanese women with breast cancer. PMID- 16378194 TI - Cyanoacrylate for wound closure in prosthetic vascular graft surgery to prevent infections through contamination. AB - PURPOSE: Prosthetic vascular graft infection (PVGI) still occurs at an unacceptably high rate, despite the many measures that have proven effective against infection. The major cause of PVGI is perioperative contamination, which usually occurs before a histological barrier is completed and may threaten graft viability. We assessed the efficacy of cyanoacrylate as a tissue adhesive and barrier against early contamination through the skin. METHOD: A 1 x 0.5-cm Dacron graft was placed subcutaneously under sterile conditions in 16 Sprague-Dawley rats. The skin was closed with polypropylene sutures in eight rats (group A) and with N-butyl 2-cyanoacrylate in eight rats (group B). The closed incisions were then contaminated with 75 x 10(6) cfu/ml methicillin-sensitive Staphylococcus aureus. Microbiological and histopathological assessments were done 7 days later, after the rats were killed. RESULTS: All of the graft cultures from group A (8/8) were positive for S. aureus whereas all of those from group B (0/8) were negative. This difference was significant (p < 0.001). Histological examination revealed denser inflammatory cell infiltration in group A than in group B. CONCLUSIONS: Maintaining skin integrity after closure is of the utmost importance because contamination during the first hours of the procedure may cause PVGI. Cyanoacrylate seems to be a promising molecule not only because of its bacteriostatic effects, but also because it seals the barrier effectively and immediately. PMID- 16378193 TI - Local granulocyte-macrophage colony-stimulating factor improves incisional wound healing in adriamycin-treated rats. AB - PURPOSE: Neoadjuvant treatment is often given for locally advanced malignancies; however, clinical and experimental studies have shown that some chemotherapeutic agents impair wound healing. It has been reported that granulocyte-macrophage colony-stimulating factor (GM-CSF) applied locally improves dermal wound healing. Thus, we investigated the effects of locally injected GM-CSF on abdominal wounds impaired by adriamycin, a widely used chemotherapeutic agent. METHODS: We divided 120 female Sprague-Dawley rats into five treatment groups of 24 rats. Group 1 received saline 8 mg/kg intravenously (i.v.) + laparotomy 14 days later (control); group 2 received 8 mg/kg i.v. adriamycin + laparotomy 14 days later; group 3 received adriamycin 8 mg/kg i.v. + laparotomy + local GM-CSF 50 microg 14 days later; group 4 received saline 8 mg/kg i.v. + laparotomy + local GM-CSF 50 microg 14 days later; and group 5 received adriamycin 8 mg/kg i.v. + laparotomy + systemic GM-CSF 50 microg 14 days later. Sutures were removed on postoperative day (POD) 7 in all five groups, and the abdominal bursting pressures were measured and recorded. Tissue samples were taken from the incision line for histopathological evaluation and hydroxyproline content measurement. RESULTS: The bursting pressure was significantly lower in groups 2 and 5 than in groups 1, 3, and 4. The hydroxyproline content and histopathological findings supported this result. CONCLUSION: The local injection of GM-CSF improved impaired wound healing in adriamycin-treated rats. PMID- 16378195 TI - Intraperitoneal versus intravenous CPT-11 given intra- and postoperatively for peritoneal carcinomatosis in a rat model. AB - PURPOSE: Peritoneal recurrence after resection of colorectal carcinoma is still a major concern. We investigated whether the novel cytostatic drug, CPT-11 (Irinotecan), delivered intraperitoneally (i.p.) and intravenously (i.v.), could inhibit intraperitoneal tumor spread in a rat model. METHODS: We induced intraperitoneal tumor growth using a tumor cell transfer model (10(6) cells) and divided the rats into the following five groups of eight: group IP1, given CPT-11 i.p. immediately after intraperitoneal tumor cell transfer; group IV1, given CPT 11 i.v. immediately after intraperitoneal tumor cell transfer; group IP2, given CPT-11 i.p. on postoperative days (PODs) 5, 10, and 15; group IV2, given CPT-11 on PODs 5, 10, and 15; and a control group. The rats were killed 30 days after tumor cell transfer, and the tumor weight, number of nodes in the greater omentum and peritoneum, presence of metastases in the liver and lungs, and ascites volume were determined. RESULTS: CPT-11 inhibited peritoneal tumor growth significantly. The direct intraoperative intraperitoneal application induced a more pronounced effect than the early postoperative intraperitoneal application, but both these application modes were superior to the intravenous route, which had no significant effect. CONCLUSION: CPT-11 was highly efficacious against peritoneal carcinomatosis in this experimental model. The combination of CPT-11 with other cytostatic agents and drugs generating different effector mechanisms may diminish or even prevent intraperitoneal tumor growth. PMID- 16378196 TI - Anaplastic thyroid carcinoma associated with granulocyte colony-stimulating factor: report of a case. AB - A 75-year-old woman was hospitalized due to a right axillary mass. She had undergone a resection of thyroid carcinoma 13 years earlier, followed by two subsequent operations for recurrent thyroid disease. A physical examination revealed a right axillary mass associated with skin ulceration. Persistent bleeding was observed at the skin ulcer associated with the right axillary lymph node, despite conservative treatment for the lesion. Surgery was thus performed to control persistent bleeding from the axillary ulcer, and a histopathological examination resulted in a diagnosis of poorly differentiated thyroid carcinoma. The postoperative course was uneventful, but marked leukocytosis and extensive skin metastases were recognized 30 days postoperatively. A systemic examination revealed no other lesions associated with marked leukocytosis, but elevated levels of granulocyte colony-stimulating factor were noted in a blood examination. As a result, her general condition deteriorated rapidly and the patient died 2 weeks after the onset of leukocytosis. PMID- 16378197 TI - Basaloid carcinoma of the thymus: report of a case. AB - We report a case of basaloid carcinoma of the thymus, confirmed by histopathological examination, in a 52-year-old man who underwent complete resection. Local recurrence and lung metastases developed, which were obliterated by radiation therapy. The patient survived for 81 months after surgery. We reviewed the 13 other cases of basaloid carcinoma of the thymus reported in the literature. Presumably, this tumor is radiosensitive and its metastatic route is blood-borne. PMID- 16378198 TI - Synchronous bilateral breast carcinoma in a 50-year-old man with 45,X/46,XY mosaic karyotype: report of a case. AB - We report a case of synchronous bilateral breast cancer in a patient with ambiguous external genitalia attributed to a 45,X/46,XY mosaicism. To our knowledge, this represents the first such case ever to be reported. Mammography, ultrasonography, computed tomography, and magnetic resonance imaging all showed bilateral suspicious breast masses with microcalcifications. There were no radiological findings of muscle invasion or axillary lymphadenopathy. The patient was successfully treated by bilateral radical modified mastectomy followed by external irradiation and adjuvant endocrine therapy. Histological examination revealed a bilateral ductal carcinoma in situ, with a cribriform and papillary pattern and microfoci of infiltrating ductal carcinoma. The hormonal profile revealed high levels of follicle-stimulating hormone and luteinizing hormone, and low levels of testosterone. Testicular sonography revealed small hypoechoic testicles with bilateral microlithiasis. This case shows that 45,X/46,XY men may have an increased risk of breast cancer and must be followed up carefully. PMID- 16378199 TI - Venovenous extracorporeal membrane oxygenation as an adjunct to surgery for empyema: report of a case. AB - Empyema is a serious and often life-threatening condition. We report the case of a 45-year-old man with severe hypoxia caused by empyema after streptococcal pneumonia, which we successfully treated by surgical drainage assisted by a venovenous extracorporeal membrane oxygenation (ECMO) device. This device provided sufficient oxygenation during the perioperative period and was not associated with excessive bleeding under systemic heparinization. This case report shows the effectiveness of ECMO in allowing surgery and enhancing its benefits in patients with serious hypoxia. PMID- 16378200 TI - Complete extirpation of a bronchogenic cyst causing recurrent laryngeal nerve palsy by thoracoscopy: report of a case. AB - We excised a bronchogenic cyst causing recurrent laryngeal nerve palsy using thoracoscopic surgery. A 28-year-old woman presented after the sudden onset of hoarseness, and laryngoscopic examination showed left vocal cord palsy. Computed tomography and magnetic resonance imaging showed a cystic mass, 4 cm in diameter, in the aortopulmonary window. Thoracoscopic examination revealed that the mass was adhered to the recurrent laryngeal nerve below the aortic arch. We extirpated the cyst via thoracoscopy without any injury to the nerves or major blood vessels. This case illustrates the benefits of thoracoscopic surgery for providing good visualization of the perineural structures and as a safe surgical treatment for a cystic mass in the aortopulmonary window. PMID- 16378201 TI - Massive hemorrhage caused by major pulmonary vessel-cutaneous fistula in the late postoperative period: report of two cases. AB - Sudden life-threatening hemorrhage caused by erosion of the wall of a thoracic blood vessel such as the aorta, pulmonary artery, or pulmonary vein, in the late postoperative period is extremely rare and presents a challenging emergency. We report the cases of two patients whose only clinical manifestation was a hemorrhagic cutaneous chest wall fistula. Both patients were treated by emergency surgery. The diagnosis and management of this clinical entity requires a high index of suspicion and innovative therapeutic solutions. PMID- 16378202 TI - Double intussusceptions in the small intestine caused by metastatic renal cell carcinoma: report of a case. AB - Renal cell carcinoma (RCC) may metastasize to almost any organ, but it is unlikely to be a direct cause of intussusception. We report a case of double enteric intussusceptions caused by metastatic RCC in a 64-year-old man. The patient presented with intermittent abdominal pain and diarrhea 11 years after undergoing a radical nephrectomy. Abdominal computed tomography (CT) showed two enhanced masses with the "target" sign, suggesting enteric intussusceptions. We performed partial enterectomy, and histological examination confirmed that the tumors had originated from RCC. To our knowledge, this is the first report of metastases from RCC manifesting as synchronous intraluminal polypoid tumors serving as the lead points of two intussusceptions in the small intestine. Thus, the possibility of multiple tumor metastases in the small intestine, with or without intussusceptions, should be considered in patients with recurrent RCC. PMID- 16378203 TI - Successful cystic fenestration for a macrocystic serous cystadenoma of the pancreas causing obstructive jaundice: report of a case. AB - A 72-year-old man was admitted to our hospital for investigation of jaundice. We made a preoperative diagnosis of macrocystic serous cystadenoma (SCA) of the pancreas, but did not perform palliative choledochojejunostomy because aspiration of the pancreatic cystic fluid caused the cysts to shrink and relieved the compressive stenosis of the common bile duct (CBD) during the operation. Frozen sections of the cyst wall taken by incision biopsy showed no signs of malignancy. Therefore, we performed fenestration of the cystic wall after fixing the inner epithelium of the cyst with 100% ethanol and aspirating the cystic fluid. Cholangiography after the cystic fenestration showed resolution of the CBD stenosis and abdominal computed tomography (CT) confirmed the disappearance of the pancreatic cysts. No recurrence of cystic swelling or obstructive jaundice has been detected by abdominal CT or laboratory data for more than 2 years since the cystic fenestration. Thus, cystic fenestration may be a better palliative option for treating benign compressive tumors such as macrocystic SCA of the pancreas causing obstructive jaundice. PMID- 16378204 TI - Laparoscopic bilateral partial adrenalectomy for adrenocortical adenomas causing Cushing's syndrome: report of a case. AB - Laparoscopic total adrenalectomy has become a standard technique for small adrenal tumors; however, bilateral adrenalectomy results in postoperative adrenal insufficiency, necessitating lifelong steroid replacement. To preserve adrenocortical function in a 41-year-old woman with bilateral adrenocortical adenoma (BAA) causing Cushing's syndrome, we performed laparoscopic bilateral partial adrenalectomy. We based our preoperative diagnosis of bilateral adrenocortical tumors causing Cushing's syndrome on the results of endocrinological investigations and imaging findings. Thus, we performed lateral transperitoneal laparoscopic bilateral partial adrenalectomy, preserving the adrenal glands, which were normal. Pathological examination of both tumors confirmed the diagnosis of adrenocortical adenoma. The patient had no postoperative complications, and her adrenocortical function was normal without steroid replacement at her 10-month follow-up. This report shows that Cushing's syndrome resulting from bilateral adenomas can be effectively treated by laparoscopic bilateral partial adrenalectomy as a minimally invasive, adrenocortical-preserving operation. PMID- 16378205 TI - Utility of an atraumatic self-retaining ring retractor in pediatric surgery. AB - The Protractor is a self-retaining ring retractor used mainly for minilaparotomies in adults. We report our positive results of using this retractor in pediatric surgery. We performed surgery with the aid of the Protractor in 57 pediatric patients aged from 1 day old to 16 years old. The Protractor allowed a wide operative view and did not cause any major complications. It was especially useful for Kasai's portoenterostomies and ureteroneocystostomy (Cohen's repair). In neonatal surgery, the Protractor not only provided a wide operative view, but also prevented the washing fluid from overflowing onto the covering sheet. In appendectomy, the Protractor protected the surgical wound from contaminated ascites and the appendix. The Protractor is a very useful tool for assisting with various abdominal operations in infants and children. PMID- 16378206 TI - A van der Waals free energy in electrolytes revisited. AB - A system of three electrolytes separated by two parallel planes is considered. Each region is described by a dielectric constant and a Coulomb fluid in the Debye-Huckel regime. In their book Dispersion Forces, Mahanty and Ninham have given the van der Waals free energy of this system. We rederive this free energy by a different method, using linear response theory and the electrostatic Maxwell stress tensor for obtaining the dispersion force. PMID- 16378207 TI - Repair of tetralogy of Fallot with anomalous coronary arteries coursing across the obstructed right ventricular outflow tract. AB - Anomalous coronary artery (ACA) anatomy occurs in 2-9% of patients with tetralogy of Fallot (TOF), in which the left anterior descending coronary artery (LAD) originates from the right coronary artery (RCA) crossing the right ventricular outflow tract. The purpose of this study was to review our results of repair for TOF with ACA. Between 1978 and 2001, 43 ACA patients (mean age, 4.8 years; range, 5 months-41 years) underwent repair for TC. The ACA anatomy was classified as the single LAD from the RCA (n = 20), a significant conal branch (dual LAD) from the RCA (n = 13), paired anterior descending arteries originating from the left and right coronary arteries (n = 7), and single RCA from the LAD (n = 3). In cases in which the anomalous LAD crossed the obstructed infundibulum, thinning or coring of the endocardium was done. Patch infundibuloplasty was performed in 39 patients, with 10 needing separate RV and pulmonary artery patches, and the pulmonary valve was preserved. Nine patients had the addition of a monocusp to a transannular incision. Two patients had a main pulmonary arterioplasty alone. There was one early (2.3%) and no late deaths. Mean early and late postoperative gradients were 21.5 +/- 10.5 mmHg (4 patients had > or =30 mmHg) and 27.1 +/- 13.7 mmHg (5 patients had >30 mmHg; p = 0.12), respectively. There were four reoperations during a mean follow-up of 4.8 years (range, 6 months to 18 years). Actuarial freedom from reoperation was 90% at 5, 10, and 15 years. At the latest follow-up, all patients were in NYHA functional class I. TOF repair for an ACA can be performed without disturbing the native coronary anatomy and without the use of conduits in most cases. Outcomes are similar to those of other patients with TOF. The presence of ACA does not impose increased risk after this surgical strategy. PMID- 16378208 TI - Follow-up assessment of the collateral circulation in patients with Kawasaki disease who underwent dipyridamole stress technetium-99m tetrofosmin scintigraphy. AB - To assess the alteration of myocardial ischemic findings and the role of collateral vessels in patients with Kawasaki disease (KD), we used dipyridamole stress technetium-99m tetrofosmin (Tf) single photon emission computed tomography (SPECT). A comparison study of coronary angiography and dipyridamole stress (0.70 mg/kg) Tf-SPECT was repeated at least twice in 20 patients. The subjects included 7 patients with coronary stenosis, 1 with pre- and post-coronary artery bypass grafting (CABG) due to coronary stenosis, 1 with progression to coronary stenosis, and 11 with persistent coronary aneurysms. In the stenosis group, Tf SPECT revealed that 6 of the 7 patients had some degree of ischemic findings, and 5 of these 6 did not show any change in their ischemic findings during follow-up. In 1 patient, the ischemic findings changed according to the collateral circulation changes. The patient who underwent CABG had pre-CABG ischemic changes that disappeared after CABG. In the patient whose coronary arteries progressed to stenosis, the ischemic findings progressed as the coronary stenosis progressed. In the persistent aneurysm group, there were no ischemic findings. In the future Tf-SPECT may become one of the most useful methods for monitoring the progressive changes of myocardial ischemia in KD. PMID- 16378209 TI - Left coronary artery arising from the non-coronary sinus: a rare congenital coronary anomaly. AB - Anomalous origin of the left coronary artery is a cause of sudden death in adolescents and young adults. Most commonly, it originates from the right coronary sinus or the pulmonary artery. Origin of the left main coronary artery from the non-coronary sinus of Valsalva is extremely rare. We report a case of a child with anomalous origin of the left main coronary artery from the non coronary sinus diagnosed during the evaluation of a ventricular arrhythmia. PMID- 16378210 TI - Nonsurgical treatment of patent arterial duct in term neonates with congenital heart disease: the role of intravenous indomethacin. AB - We present two term neonates in which intravenous indomethacin usage was associated with successful closure of a persistently patent arterial duct. Both children had congenital heart disease with high pulmonary blood flow following surgical placement of contralateral systemic-pulmonary artery shunts. PMID- 16378211 TI - Transcatheter closure of a large patent ductus arteriosus in a young child using the Amplatzer duct occluder. AB - The Amplatzer duct occluder (ADO) provides a safe and effective therapy for patients with moderate- to large-sized patent ductus arteriosus (PDA), but there have been few reports of transcatheter closure of very large PDAs in young children and infants. We report a successful transcatheter closure of a very large PDA, 10.5 mm in diameter at the narrowest point, with a 14/12-mm ADO. To our knowledge, this is the largest PDA ever closed by an interventional method in such a young child. PMID- 16378212 TI - Digital area measurement of fundus landmarks. AB - PURPOSE: To present and validate an image analysis technique for the calculation of the true projection area of fundus landmarks from a digital fundus image. The authors present the description, technical details and results of the study. METHODS: Experimental study using a calibrated Gullstrand-type model eye with a scale of half circles in the centre of the artificial fundus. The axial length of the model eye was set to different values ranging from 21 mm to 29 mm, and the area of a half circle was calculated from digital fundus images with a magnification-calibrated image analysis program (UTHSCSA Image Tool). The calibration was based on Littmann's formula for correcting the magnification of fundus images. RESULTS: The maximum deviation of the calculated area size from the true fundus object area was -3.7% under myopic and +3.4% under hyperopic conditions. The 95% confidence interval for repeated area measurements was from +1.83 to -1.46%. CONCLUSIONS: The magnification-calibrated image analysis technique based on Littmann's formula is a non-invasive, accurate and reproducible tool for true area measurement of fundus landmarks. PMID- 16378213 TI - Heparan sulfate proteoglycans mediate Staphylococcus aureus interactions with intestinal epithelium. AB - Staphylococcus aureus can be internalized by non-professional phagocytes, and may colonize the intestine in normal and antibiotic-treated individuals. Intestinal colonization may depend on the interactions of S. aureus with the intestinal epithelium. The best described mechanism of S. aureus binding to eukaryotic cells involves S. aureus fibronectin binding proteins (FnBPs), using fibronectin as a bridging molecule to beta1 integrins on the eukaryotic cell surface. Because S. aureus can be internalized by enterocytes, and because S. aureus is known to bind heparan sulfate (HS), we hypothesized that heparan sulfate proteoglycans (HSPGs) widely expressed on epithelia may mediate S. aureus interactions with intestinal epithelial cells. Internalization of S. aureus RN6390 by cultured intestinal epithelial cells was inhibited in a dose-dependent fashion by the HS mimic heparin, and by HS itself. Internalization of S. aureus DU5883, which lacks expression of staphylococcal FnBPs, was also inhibited by heparin. S. aureus adherence to ARH-77 cells, transfected to express the HSPG syndecan-1, was greatly increased when compared to adherence to plasmid control ARH-77 cells which have little detergent extractable HS. In addition, compared to wild-type HS expressing Chinese hamster ovary (CHO) cells, internalization of S. aureus was decreased using mutant CHO cells with decreased HS expression. These findings are consistent with a model wherein S. aureus internalization by intestinal epithelial cells (and perhaps other epithelia) is mediated by S. aureus binding to the HS moiety of cell-surface HSPGs, and this interaction appears independent of fibronectin binding. PMID- 16378214 TI - [The Third IAS-Conference on HIV-Pathogenesis und Treatment, Rio de Janeiro, 24 27 July, 2005, Part 1]. PMID- 16378215 TI - 3,4-Methylenedioxymethamphetamine enhances the release of acetylcholine in the prefrontal cortex and dorsal hippocampus of the rat. AB - RATIONALE: The neurochemical effects produced by acute administration of 3,4 methylenedioxymethamphetamine (MDMA) on the monoaminergic systems in the brain are well documented; however, there has been little consideration of the potential effects of MDMA on other neurotransmitter systems. OBJECTIVE: The present study was designed to investigate the acute effect of MDMA on cholinergic neurons by measuring acetylcholine (ACh) release in the medial prefrontal cortex (PFC) and dorsal hippocampus, terminal regions of cholinergic projection neurons originating in the basal forebrain. METHODS: In vivo microdialysis and high performance liquid chromatography with electrochemical detection (HPLC-ED) were used to assess the effects of MDMA on the extracellular concentration of ACh in the PFC and dorsal hippocampus of the rat. RESULTS: The systemic administration of MDMA (3-20 mg/kg, i.p.) resulted in an increased extracellular concentration of ACh in the PFC and dorsal hippocampus. Reverse dialysis of MDMA (100 microM) into the PFC and hippocampus also increased ACh release in these brain regions. Treatment with parachlorophenylalanine and alpha-methyl-para-tyrosine, inhibitors of serotonin (5-HT) and dopamine (DA) synthesis, respectively, significantly attenuated the release of ACh stimulated by MDMA in the PFC, but not in the dorsal hippocampus. CONCLUSIONS: MDMA exerts a stimulatory effect on the release of ACh in the PFC and dorsal hippocampus in vivo, possibly by mechanisms localized within these brain regions. In addition, these results suggest that the MDMA-induced release of ACh in the PFC involves both serotonergic and dopaminergic mechanisms. PMID- 16378216 TI - The origin of pharmacopsychology: Emil Kraepelin's experiments in Leipzig, Dorpat and Heidelberg (1882-1892). AB - This historical review shows that the early history of cognitive psychopharmacology, originally labelled as "pharmacopsychology", is closely linked to developments in experimental psychology and academic psychiatry. At the beginning of his scientific career, the German psychiatrist Emil Kraepelin (1856 1926) joined Wilhelm Wundt's laboratory of experimental psychology at the University of Leipzig. Although Kraepelin was fired from his clinical position at the university's psychiatric hospital, he completed his habilitation, the German equivalent of Ph.D., and started a series of pharmacological investigations in healthy volunteers using common recreational drugs (alcohol, coffee, tea) or medicinal products (amyl nitrite, chloral hydrate, chloroform, ethyl ether, morphine, paraldehyde) together with innovative psychological tasks. This paper reviews Kraepelin's pharmacopsychological research and his methodological innovations, providing translations, for the first time, from original papers, his monograph On the Modulation of Simple Psychological Processes by Some Medicines and from other sources. Kraepelin's contributions to psychopharmacology and clinical neuropsychology were far ahead of his time and his conceptual achievements have been largely neglected by modern psychiatry and cognitive neuroscience. PMID- 16378217 TI - Anti-conflict effects of benzodiazepines in rhesus monkeys: relationship with therapeutic doses in humans and role of GABAA receptors. AB - RATIONALE AND OBJECTIVES: Conflict procedures are used to study mechanisms underlying the anxiolytic effects of benzodiazepines (BZs). We established a conflict procedure with rhesus monkeys in order to examine the role of GABAA receptors in the anxiolytic-like effects of BZs. METHODS: Four rhesus monkeys responded under a two-component multiple schedule in which responding was maintained under a fixed-ratio schedule of food delivery in the absence (non suppressed responding) and presence (suppressed responding) of response contingent electric shock. RESULTS: Conventional BZs (alprazolam, flunitrazepam, clonazepam, nitrazepam, lorazepam, bromazepam, diazepam, flurazepam, clorazepate, chlordiazepoxide) engendered increases in the average rates of suppressed responding at low to intermediate doses and decreased the average rates of non suppressed responding at higher doses. Positive correlations were observed when the therapeutic potencies of BZs in humans were compared with potencies to increase the rates of suppressed responding (R2=0.83) or decrease the rates of non-suppressed responding (R2=0.60). The 5-HT1A agonist buspirone increased the rates of suppressed responding, although the effects were modest, whereas the opioid morphine lacked anti-conflict effects. The BZ antagonist flumazenil also modestly increased the rates of suppressed responding. A relatively low dose of flumazenil enhanced, while a high dose blocked, alprazolam's anti-conflict effects. Compounds selective for alpha1 subunit-containing GABAA receptors (zolpidem, zaleplon, CL218,872) engendered relatively weak increases in the rates of suppressed responding. CONCLUSIONS: A rhesus monkey conflict procedure was established with predictive validity for therapeutic doses in people and provided evidence that anxiolytic-like effects of BZs can occur with relatively low intrinsic efficacy at GABAA receptors and are reduced by alpha1GABAA receptor selectivity. PMID- 16378219 TI - Relationships between local and regional species richness in flea communities of small mammalian hosts: saturation and spatial scale. AB - The number of species coexisting in a community may be regulated by local factors (e.g., competitive interactions), or by regional processes (e.g., dispersal from a regional species pool). The relative importance of local and regional processes can be inferred from the shape of the relationship between local and regional species richness. We investigated this relationship in communities of fleas parasitic on small mammals at two spatial scales: between the richness of fleas on individual hosts (infracommunities) and that of fleas on host populations (component communities), and between the richness of component communities and that of the entire regional species pool. We tested linearity (proportional sampling) versus curvilinearity with an asymptote (species saturation) by plotting "local" against "regional" species richness of fleas either among host species or within host species among populations. At the two spatial scales, we found consistent curvilinear relationships between species richness of the more "local" communities and richness of the more "regional" communities. This was true across all host species in the data set and for geographic subsets, even after controlling for the influence of sampling effort on estimates of species richness, and that of host phylogeny in interspecific analyses. We also tested for density compensation in species-poor communities. There was no strong evidence for density compensation at the infracommunity level, although its existence at the component community level appeared likely. Our results suggest that identical patterns in local-versus-regional species richness observed on two different spatial scales arise via different mechanisms: infracommunities appear saturated with flea species most likely because of local processes, such as host immune defenses, whereas component communities are saturated with species through interspecific competition, possibly among larval stages. PMID- 16378218 TI - Alcohol-induced impulsivity in rats: an effect of cue salience? AB - RATIONALE: There is a common assumption that alcohol produces impulsive behaviors, thereby increasing the preference for immediate over delayed rewards. An alternative explanation, provided by alcohol myopia theory, is that alcohol alters attentional processes such that intoxicated individuals respond exclusively to the most salient cues in their environment. OBJECTIVES: We tested these two hypotheses in rats using standard (impulsivity) and modified (cue salience) versions of the delayed reinforcement task. METHODS: In the impulsivity paradigm, rats were trained to choose between a small immediate reward (2 sucrose pellets) and a large delayed reward (12 sucrose pellets after 10 s) in a T-maze. In the cue salience paradigm, a light in one arm predicted either the small or the large reward, but the arm paired with the light varied across trials. In separate experiments, we examined how changes in delay to the large reward, alcohol administration, or alcohol combined with either a serotonin agonist [para chloroamphetamine (pCA) at doses of 0.1, 0.4, 0.7, and 1.0 mg/kg, s.c.)] or a dopamine antagonist [cis-(Z)-flupenthixol dihydrochloride (alpha-flupenthixol) at doses of 0.0125, 0.025, 0.05, and 0.1 mg/kg, i.p.] affected performance in each task. RESULTS: Increasing the delay to the large reward increased impulsivity in both paradigms, but it had no effect on responding to a salient cue. Alcohol (0.6, 0.9, 1.2, and 1.8 g/kg, i.p.) increased the choice of the immediate reward and increased the choice of the lit arm, regardless of whether it signaled the small or the large reward. pCA selectively reduced alcohol-induced impulsivity, whereas alpha-flupenthixol selectively reduced responding to a salient cue. CONCLUSIONS: Rats, like humans, are influenced by cue salience when intoxicated. Although this alcohol myopia effect could explain alcohol-induced impulsivity, the two processes are probably distinct because they are mediated by dissociable pharmacological mechanisms. PMID- 16378220 TI - In vitro effects of Crotalus durissus terrificus and Bothrops jararaca venoms on Giardia duodenalis trophozoites. AB - Considering the snake venoms' pharmacological properties and chemotherapeutic potential as well as the need for new alternatives for Giardia infection treatment, the present study was carried out aiming to evaluate the in vitro effects of crude Crotalus durissus terrificus and Bothrops jararaca venoms on the growth and adherence of Giardia duodenalis trophozoites. Trophozoites (10(6)) were exposed to serial twofold dilutions of C. durissus terrificus and B. jararaca venoms that ranged from 3.125 to 200 microg/ml and from 5 to 320 microg/ml, respectively. The two venoms inhibited the growth of trophozoites, and the level of inhibition varied according to the assayed concentrations and incubation times. The highest reduction of parasite growth was observed with increasing concentrations of the venoms. No effect on parasite adherence was observed. Light microscope observations revealed changes of the pear-shape aspect of the cell and reduction of flagellar beating frequency in the great part of the trophozoites. This is the first attempt to evaluate the in vitro effects of snake venoms on Giardia trophozoites. The findings stress the need for more investigations and prompt us to perform further studies to identify active principles with antigiardial effect. PMID- 16378221 TI - Necrotizing external otitis in a patient caused by Klebsiella pneumoniae. AB - Necrotizing external otitis is a potentially life-threatening infection involving the temporal and adjacent bones. The most frequent pathogen is attributed to Pseudomonas aeruginosa, but is rarely caused by Klebsiella pneumoniae. Recently, we encountered a 47-year-old diabetic man with a swollen obliterated external ear canal with granulation tissue on the right ear. Image study demonstrated skull base osteomyelitis, epidural abscess and cerebral venous sinus thrombi. It was later proved to be necrotizing external otitis caused by Klebsiella pneumoniae. He then underwent craniotomy for drainage of the epidural abscess, followed by intravenous ciprofloxacin and metronidazole for 2 consecutive weeks until both pus and blood cultures depicted no growth of pathogens. Based on this case, synergistic antibiotic therapy using a third-generation cephalosporin or quinolone (ciprofloxacin), accompanied by metronidazole, and even a short-term aminoglycoside is recommended for the treatment of severe Klebsiella-induced necrotizing external otitis. Surgical intervention should be limited without shedding of the pathogens. PMID- 16378222 TI - Accessory or sublingual salivary gland biopsy to assess systemic disease: a comparative retrospective study. AB - Minor salivary gland biopsies are commonly performed as part of the diagnosis of Sjogren's syndrome or other systemic diseases. Until now, a lip biopsy taken from inside the inferior lip has been the most often performed method to assess the accessory salivary glands. Because of the risk of damaging the inferior alveolar nerve and of harvesting non-contributive biopsies, for the past several years we have chosen the sublingual biopsy described by Adam. The aim of this study was to describe the indication and diagnosis and to evaluate work incapacity, pain and the complication rate among our salivary gland biopsies. In this retrospective study, we evaluated 79 biopsies (lip, n=24, and sublingual, n=55) taken at the Department of Oral and Maxillofacial Surgery of the University Hospital St-Luc, Brussels, by multiple junior trainees and senior surgeons (n=17). Data were collected from record study and from telephonic patient questionnaires. The three major indications were: sicca syndrome (24%), suspicion of Sjogren's syndrome (32%) or exclusion of other systemic diseases (44%). The histology results of the lip and sublingual biopsies, respectively, were: normal tissue (29%, 24%), chronic inflammation (29%, 41%), compatible, but not characteristic for Sjogren (8%, 7%), Sjogren's syndrome (13%, 24%), salivary gland atrophy (13%, 0%) or non contributive (8%, 0%). The procedures were both almost painless (time during which painkillers had to be taken: 0.33 vs. 0.69 days, NS) and work incapacity was not encountered in any group. After one lip biopsy we had to deal with a permanent anesthesia of the lower lip (6.6%), and after one sublingual biopsy a swelling of the floor of the mouth had to be incised under local anesthesia (2.7%). Thus, sublingual biopsy is an easy procedure with low morbidity and excellent reliability in comparison to lip biopsy. No salivary gland atrophy and no non-contributive biopsies were encountered; no important bleeding or nerve lesion was recorded after sublingual biopsies. PMID- 16378223 TI - Apoptosis in the pathogenesis of cholesteatoma in adults. AB - The aim of this study was to determine the distribution and apoptotic capacity in the various layers of cholesteatoma epithelium compared to the normal skin. Cholesteatomas were collected during surgical procedures of the ear. Normal skin specimens taken from the retroauricular area served as controls. Apoptosis was detected by using highly specific antibodies against APO2.7 antigen. The Membstain Apoptosis kit Direct based on in situ labeling of nuclear DNA fragmentation (Tolt-mediated dUTP nick and labeling TUNEL staining) was used. The distribution of apoptotic cells in the layer of the cholesteatoma epithelium and the epidermis was determined. The percentage of apoptotic cells was then counted per 100 cells of cholesteatoma epithelium and the normal epidermis in three different areas of each section and expressed in terms of mean +/- SD. An automatic analyzing system was used for counting. In normal epidermis, the apoptotic cells were observed in the granular layer of the epidermis. The percentage of these cells was 28.5 +/- 8.1%. A significantly greater number of apoptotic cells was observed in the suprabasal layers of the cholesteatoma epithelium (47.39 +/- 6.2%). A statistically significant difference was obtained for APO2.7-positive cells in the cholesteatoma epithelium as compared to the skin (P < 0.05) as determined by Student's t-test and the Mann-Whitney U test. Apoptotic activity is mainly observed in the suprabasal layer of cholesteatoma epithelium. It is not seen in the basal layer. Apoptosis plays an important role in the pathogenesis of cholesteatoma. It seems to be part of the differentiation and accumulation of keratin debris within the middle ear and expansion of cholesteatoma. PMID- 16378224 TI - Molecular targets of cancer chemotherapy. PMID- 16378225 TI - Short-term dexamethasone treatment. PMID- 16378226 TI - Validity of teacher ratings in selecting influential aggressive adolescents for a targeted preventive intervention. AB - This study describes a method for using teacher nominations and ratings to identify socially influential, aggressive middle school students for participation in a targeted violence prevention intervention. The teacher nomination method is compared with peer nominations of aggression and influence to obtain validity evidence. Participants were urban, predominantly African American and Latino sixth-grade students who were involved in a pilot study for a large multi-site violence prevention project. Convergent validity was suggested by the high correlation of teacher ratings of peer influence and peer nominations of social influence. The teacher ratings of influence demonstrated acceptable sensitivity and specificity when predicting peer nominations of influence among the most aggressive children. Results are discussed in terms of the application of teacher nominations and ratings in large trials and full implementation of targeted prevention programs. PMID- 16378227 TI - Development of cell therapy using autologous bone marrow cells for liver cirrhosis. AB - The plasticity of bone marrow has been confirmed by the autopsy of a female recipient of bone marrow cell transplantation from a male donor. To establish new clinical cell therapies using autologous bone marrow cells for patients with liver failure, we developed a new in vivo model named the green fluorescent protein (GFP)/carbon tetrachloride (CCl4) model. Using the GFP/CCl4 model, we found that transplanted Liv8-negative cells efficiently repopulated into cirrhotic liver tissue and differentiated into albumin-producing hepatocytes under persistent liver damage induced by carbon tetrachloride. Moreover, bone marrow cell transplantation into mice with liver cirrhosis improved liver function and liver fibrosis with the strong expression of matrix metalloproteinases (MMPs), especially MMP-9 activity, resulting in an improved survival rate. Results from the GFP/CCl4 model showed that cell therapy using autologous bone marrow cells has the potential to become an effective treatment for patients with liver failure. A summary of findings from the GFP/CCl4 model is described. PMID- 16378229 TI - MRI of the breast: comparison of MRI signals and histological characteristics of the same slices. AB - The development of MRI with high spatial resolution and a special breast coil has contributed to more-accurate diagnosis of breast tumor, such as determination of morphologic characteristics including internal architecture of the breast lesion. To clarify how individual MRI findings reflect the pathological findings, we made slices of resected breast lesions identical to those obtained with axial MRI and compared them respectively according to histological subtype of tumors. In this article, we present MRI findings on the basis of histopathological evidence. In general, fibroadenomas (FA) show a well-defined border on contrast-enhanced fat suppressed T1-weighted images (CFT1). However, a subtype of fibroadenoma, mastopathic type, with a poorly defined border on CFT1 and carcinoma-like enhancement, is similar to cancer on MRI. Scirrhous carcinomas take either of two patterns on dynamic MRI, i.e., homogeneous enhancement or thick irregular peripheral ring enhancement. The latter type shows central fibrosis zones histologically. Papillotubular carcinoma shows a spotted pattern on dynamic MRI. The low-enhancement areas within the spotted pattern reflect parenchyma between clusters of cancer. Solid-tubular carcinoma shows thin peripheral enhancement on dynamic MRI and linear high signal on CFT1. This finding of CFT1 corresponds to infiltration of lymph cells and fibroblast cells in the adjacent zone. Invasive lobular carcinoma shows nonmass lesions and a slow gradual enhancement pattern. Mucinous carcinomas show high signal intensity on T2-weighted images because of mucin and reveal gradual enhancement. The ill-defined border of mucinous carcinoma on CFT1 is useful to distinguish it from FA. PMID- 16378230 TI - Electron microscopic findings for diagnosis of breast lesions. AB - The normal mammary gland can be roughly divided into the large duct close to the nipple and the terminal duct located within the lobulus. Both the large duct and the terminal duct are composed of epithelial cells and myoepithelial cells. The epithelial cells can be divided into light and dark cells using electron density. Heterochromatin is the predominant type of chromatin found in normal mammary glands. The cytoplasm of myoepithelial cells contains a number of fine filaments that possess dense patches. The myoepithelial cells of the large duct have a large process with a crablike appearance that protrudes from the cytoplasm. The myoepithelial cells of the terminal duct, by contrast, assume a relatively flat form and are approximately parallel to the epithelial-stromal junction. If the nuclei of the epithelial cells of normal mammary glands and benign breast lesions are compared with those of malignant breast lesions, the latter are primarily oval or circular in shape whereas the former often show marked notches. The predominant chromatin is heterochromatin in noncancer cells and euchromatin in cancer cells. The intracytoplasmic lumen (ICL) can be roughly divided into two types. The ICL is frequently seen in breast cancers, especially scirrhous carcinoma and lobular carcinoma. Invasive ductal carcinoma can be divided into three types: papillotubular carcinoma, solid-tubular carcinoma, and scirrhous carcinoma. Scirrhous carcinoma can be divided into two subtypes: scirrhous carcinoma in the broader sense of the term (characterized by scirrhous invasion of the stroma by papillotubular carcinoma or solid-tubular carcinoma), and scirrhous carcinoma in the narrower sense of the term (characterized by linear or cluster-like invasion of the stroma without forming ducts). Ultrastructural characteristics of scirrhous carcinoma in the narrow sense are bright cytoplasm (seen in most cells) and euchromatin (seen in all cells of this type of carcinoma). In cases of papillotubular carcinoma, solid-tubular carcinoma, and scirrhous carcinoma in the broad sense, euchromatin is predominant but sporadic cells with heterochromatin are also seen. Adenoid cystic carcinoma and carcinoid tumor of the breast are histological types of breast carcinoma that show characteristic features under an electron microscope. Ultrastructurally, the former shows a pseudocyst and true lumen whereas the latter presents numerous neuroendocrine granules within the cytoplasm. Breast carcinoma shows several ultrastructural characteristics that are useful in differential diagnosis. Therefore, it is advisable to take electron microscopic findings into account when evaluating or diagnosing breast lesions. PMID- 16378231 TI - Long-term inhibition of intimal hyperplasia using vascular photodynamic therapy in balloon-injured carotid arteries. AB - Flexible treatments for intimal hyperplasia after angioplasty are still needed. The aim of this study was to demonstrate the long-term effects of vascular photodynamic therapy with talaporfin sodium on intimal hyperplasia following interventional injury. Intimal hyperplasia was induced by balloon distension injury to the carotid artery in 31 rabbits. Talaporfin, 5.0 mg/kg, was delivered systemically immediately after balloon injury. The injury site was irradiated with a diode laser light of wavelength 664 nm using a fluence of 50 J/cm2 after 30 min. At day 3 and weeks 3, 6, 9, 15, and 25 after photodynamic therapy, the treated artery of each rabbit was excised and examined immunohistochemically. Thirty minutes after talaporfin administration, drug fluorescence was found only in the balloon-injured carotid artery wall. At 3 days, no smooth muscle cells were seen in the media of the photodynamic therapy-treated arterial segments. Intimal hyperplasia developed progressively in the balloon-injured and untreated segments; however, in the segments treated with photodynamic therapy, intimal hyperplasia was markedly suppressed until 25 weeks and the media was repopulated by smooth muscle cells without macrophages. Vascular photodynamic therapy with talaporfin may be used to inhibit restenosis after vascular intervention. PMID- 16378232 TI - An immunocytochemical study of regeneration of gastric epithelia in rat experimental ulcers. AB - We experimentally observed the process of regeneration of gastric mucosal cells in ulcers induced by surgical removal of the fundic mucosa of rats. The techniques utilized were immunocytochemistry, laser confocal microscopy, and transmission electron microscopy (TEM). Routine TEM and PAS reaction were used for parietal cells, chief cells, and mucous cells. As markers of parietal cells, H+-K+ ATPase, Na+-K+ ATPase, carbonic anhydrase, and aquaporin 4 (AQP4) were used, and for chief cells pepsinogen was used. Healing process of mucosal defect was as follows. On day 2 after the operation, the single-layered regenerating epithelium (RE) originating from the marginal epithelium of the ulcer extended over the granulation tissue of the ulcer base towards the center. Regenerating glands (RGs) appeared in the ulcer margin. The cells appearing first in the RE were undifferentiated cells that had a high nucleus: cytoplasm ratio and abundant free ribosomes. On day 5, the ulcer was almost filled with RGs. Most cells stained positive for PAS reaction. A few immature parietal cells stained weakly with H+-K+ ATPase, Na+-K+ ATPase, carbonic anhydrase, and aquaporin-4 antibodies, and a few immature chief cells stained weakly with pepsinogen antibody were also observed on day 5. On day 10, the ulcer was filled with RGs. The RGs in the periphery of the ulcer stained positive for markers of mature parietal cells and chief cells, whereas the center of the ulcer was composed of immature parietal cells and chief cells. By day 25, the mucosal defect was filled with normal gastric glands formed by maturation of the RGs. The undifferentiated cells that first appeared in the ulcer margin seem to differentiate to special functioning cells of the stomach 5-10 days after ulcer formation. PMID- 16378233 TI - Sodium bicarbonate secretion indicated by ultrastructural cytochemical localization of HCO3(-), Cl-, and Na+ ions on rat bile duct brush cells. AB - Brush cells are widely distributed in the digestive and respiratory apparatus, but their function is still unknown. Because brush cells (BC) are found in organs secreting NaHCO3, it was hypothesized that these cells may secrete NaHCO3. To test this possibility, rat common bile duct epithelia were examined by ultrastructural cytochemical methods for localizing HCO3(-), Cl-, and Na+ ions. All three ion precipitates were few in or on BCs of rats without stimulation. Lead carbonate precipitates, which localized HCO3(-) ions by the lead nitrate osmium method, increased markedly on the surface of the microvilli (MV) of BCs after secretin or meal stimulation, but similar precipitates were few on the luminal surface of principal cells (PCs). Silver chloride precipitates, which indicate the presence of Cl- ions by the silver-osmium method, increased in the apical cytoplasm and in MV of BCs after secretin or meal stimulation, but they were few in PCs. Sodium pyroantimonate precipitates, which localize Na+ ions by the potassium pyroantimonate-osmium method, increased on the surface of the MV, along the basolateral membrane, and in the apical cytoplasm of BCs after secretin or meal stimulation, but they were few in PCs. These results strongly suggest that BCs may be a significant source of NaHCO3 secretion. PMID- 16378234 TI - Toxoplasma gondii: localization of purine nucleoside phosphorylase activity in vitro and in vivo by electron microscopy. AB - Purine nucleoside phosphorylase (PNP, EC 2.4.2.1) activity was revealed by enzyme histochemistry in Toxoplasma gondii ME49 strain isolated from murine cerebral cysts and from in vitro cultivation. The activity of the enzyme was revealed by an insoluble electron-opaque precipitate of lead phosphate at the site of the reaction. In bradyzoites and tachyzoites of T. gondii, the enzyme activity could be observed only in the cytoplasm. In bradyzoites, one or two foci of important PNP activity were detected near the nucleus. In tachyzoites, an important PNP activity underlined the plasma membrane. For both bradyzoites and tachyzoites, localization neither in the nucleus nor in cytoplasmic organelles could be detected. PMID- 16378235 TI - Primary gastric small cell carcinoma: report of a case and review of the literature. AB - A 52-year-old man suffering from a pure-type primary gastric small cell carcinoma was treated with surgery and combination chemotherapy. The small cell carcinoma, approximately 6.5 cm in diameter, was situated in the posterior wall of the antrum and there were no distant metastases. Total gastrectomy and regional lymph node dissection was carried out. Histological examination revealed a solid pattern of proliferation of small cells with hyperchromatic, round nuclei and scant cytoplasm. Neoplastic cells infiltrated into the subserosal layer with severe lymphatic and vascular invasion. Regional lymph node cells were mostly replaced by tumor cells that stained positive for Grimelius, neuron-specific enolase (NSE), and synaptophysin. Accumulations of electron-dense core granules in the small neoplastic cells were seen by electron microscopy. Following surgery, the patient was treated with adjuvant chemotherapy consisting of cisplatin and etoposide. The patient is alive and recurrence free 3 years after surgical operation. We review 107 published cases of primary gastric small cell carcinoma, an extremely rare disease first reported in 1976. Small cell carcinoma is an aggressive, malignant tumor. Intensive chemotherapy is essential for patient survival even when curative surgical resection is carried out. PMID- 16378236 TI - Eosinophilic cytoplasmic inclusions in papillary renal cell carcinoma. AB - A case of a patient with papillary renal cell carcinoma (RCC) with eosinophilic inclusions is presented. Almost all neoplastic cells contained eosinophilic globules that were stained red by trichrome and were negative for periodic acid Schiff (PAS) reaction. Immunohistochemically, globules were negative for actin, keratin, vimentin, glially fibrillary acidic protein (GFAP), and neurofilament. Ultrastructurally, globules consisted of electron-dense granular material that is more suggestive of a secretory substance than cytoskeletal filaments. The presence of eosinophilic cytoplasmic inclusions in RCC is rare. In this article, we review similar cases in the literature and discuss the nature of eosinophilic globules. PMID- 16378237 TI - Medical errors: getting the incentives right. AB - This work examines the role of penalties as providers of incentives to prevent medical errors and ensure that such incidents, once they occur, become common knowledge. It is shown that a scheme with two penalties (accountability and non report) induces the first-best solution. However, this scheme does not necessarily imply a punitive environment, but may, under given circumstances, yield insignificant and even negative penalties. Alternative sanction systems, such as voluntary reporting and immunity, are found to have less desirable properties. An exception is confidentiality (anonymity) which turns out to be an optimal scheme. Finally, the examination of various penalty restrictions (scope and scale) shows that such barriers may promote both tougher and softer sanction schemes. PMID- 16378238 TI - Hospital health care: pricing and quality control in a spatial model with asymmetry of information. AB - The cost of hospital care depends on the quality of the service, on the personal characteristics of the patient, on the effort of the medical staff and on information asymmetry. In this article the cost minimizing properties of alternative payment systems will be discussed in a context where hospitals can observe patient severity and compete according to the rules of Hotelling's spatial competition. The scheme is designed from the standpoint of a purchaser that sets up a contract with several providers for services of a given quality at the least possible cost. Patients' severity cannot be observed and quality cannot be verified, but the latter can be inferred through the choice of patients. The model shows that in the health care market, prospective payments and yardstick competition are weak instruments for cost containment; incentive compatible schemes are, at least from a theoretical point of view, better instruments especially in a context where the purchaser can use signals relating to the variables it cannot observe. Cost inflation has two components: the information rent paid to the provider and inefficiency. In our model the information rent is used by the provider to get more patients to his hospital; spatial competition can then be used to curb the cost of providing hospital care. PMID- 16378239 TI - Health insurance disparities in traditional and contingent/alternative employment. AB - Relative to whites, Hispanics and blacks are less likely to have employer health insurance coverage. We examine whether ethnicity or race affects employment in traditional jobs or in contingent and alternative work arrangements, and whether ethnicity or race affects insurance offer, eligibility, and/or enrollment, conditional on employment sector. Health insurance disparities relative to whites are more pronounced for Hispanics, primarily due to disparities in employment by firms that offer coverage. Eliminating racial/ethnic disparities in offers, eligibility, and takeup would increase insurance coverage rates of Hispanics in traditional jobs and of both Hispanics and blacks in contingent and alternative jobs. PMID- 16378240 TI - Willingness-to-pay and demand curves: a comparison of results obtained using different elicitation formats. AB - Health economists use "willingness-to-pay" to assess the prospective value of novel interventions. The technique remains controversial, not least with respect to the formats under which values are elicited. The paper analyses the results of a series of studies of the same intervention valued by the same population, in which different elicitation formats were employed. The findings support the hypothesis that data collected using different formats give rise to different demand curves, from which different inferences about demand elasticity, profitability and consumer surplus will be derived. Judgements as to the relative merits of rival interventions depend crucially upon whichever format has been used to evaluate each intervention. PMID- 16378241 TI - Role of the conserved glutamine 291 in the rat gamma-aminobutyric acid transporter rGAT-1. AB - We investigated the role of the Q291 glutamine residue in the functioning of the rat gamma-aminobutyric acid (GABA) transporter GAT-1. Q291 mutants cannot transport GABA or give rise to transient, leak and transport-coupled currents even though they are targeted to the plasma membrane. Coexpression experiments of wild-type and Q291 mutants suggest that GAT-1 is a functional monomer though it requires oligomeric assembly for membrane insertion. We determined the accessibility of Q291 by investigating the impact of impermeant sulfhydryl reagents on cysteine residues engineered in close proximity to Q291. The effect of these reagents indicates that Q291 faces the external aqueous milieu. The introduction of a steric hindrance close to Q291 by means of [2 (trimethylammonium)ethyl] methanethiosulfonate bromide modification of C74A/T290C altered the affinity of the mutant for cations. Taken together, these results suggest that this irreplaceable residue is involved in the interaction with sodium or in maintaining the cation accessibility to the transporter. PMID- 16378242 TI - Multiple cAMP-induced signaling cascades regulate prolactin expression in T cells. AB - Beside its pivotal role in reproduction, the pituitary hormone prolactin (PRL) has been attributed an immunomodulatory function. Here we report that cAMP is an important stimulator of PRL transcription in primary human T lymphocytes. Inhibition of both protein kinase A (PKA) and p38 MAPK partially abrogated cAMP induced PRL expression. In addition, cAMP-induced phosphorylation of p38 was shown to occur independently of PKA and could be mimicked by a methylated cAMP analogue which specifically activates the recently discovered cAMP receptor EPAC (exchange protein directly activated by cAMP). Our findings suggest that cAMP induces PRL expression in T lymphocytes via cooperation of at least two different signaling pathways: a PKA-dependent pathway leading to the phosphorylation of cAMP response element-binding protein, and a PKA-independent pathway leading to p38 phosphorylation. PMID- 16378243 TI - Functional polymorphisms of the brain serotonin synthesizing enzyme tryptophan hydroxylase-2. AB - Many neuropsychiatric disorders are considered to be related to the dysregulation of brain serotonergic neurotransmission. Tryptophan hydroxylase-2 (TPH2) is the neuronal-specific enzyme that controls brain serotonin synthesis. There is growing genetic evidence for the possible involvement of TPH2 in serotonin related neuropsychiatric disorders; however, the degree of genetic variation in TPH2 and, in particular, its possible functional consequences remain unknown. In this short review, we will summarize some recent findings with respect to the functional analysis of TPH2. PMID- 16378244 TI - Effects of constitutively active GTPases on fibroblast behavior. AB - The GTP-binding proteins RhoA, Cdc42 and Rac1 regulate the organization and turnover of the cytoskeleton and cell-matrix adhesions, structures bridging cells to their support, and translating forces, external or generated within the cell. To investigate the specific requirements of Rho GTPases for biomechanical activities of clonal cell populations, we compared side-by-side stable lines of human fibroblasts expressing constitutively active (CA) RhoA, Cdc42 or Rac1. There was no marked effect of any CA GTPase on cell adhesion to different extracellular matrix proteins. Cell spreading was CA Rho GTPase specific and independent of the extracellular matrix proteins allowing adhesion. Mechanical properties were dramatically restricted by CA RhoA on bi- and in tri-dimensional surroundings, were boosted by CA Rac1 on bi-dimensional surroundings only, and were not or marginally affected by CA Cdc42. In conclusion, the action of Rho GTPases appears to depend on the task cells are performing. PMID- 16378245 TI - The Nudix hydrolase superfamily. AB - Nudix hydrolases are found in all classes of organism and hydrolyse a wide range of organic pyrophosphates, including nucleoside di- and triphosphates, dinucleoside and diphosphoinositol polyphosphates, nucleotide sugars and RNA caps, with varying degrees of substrate specificity. Some superfamily members, such as Escherichia coli MicrotT, have the ability to degrade potentially mutagenic, oxidised nucleotides while others control the levels of metabolic intermediates and signalling compounds. In prokaryotes and simple eukaryo tes, the number of Nudix genes varies from 0 to over 30, reflecting the metabolic complexity and adaptability of the organism. Mammals have around 24 Nudix genes, several of which encode more than one variant. This review integrates the sizeable recent literature on these proteins with information from global functional genomic studies to provide some insights into the possible roles of different superfamily members in cellular metabolism and homeostasis and to stimulate discussion and further research into this ubiquitous protein family. PMID- 16378247 TI - Isoprenylated proteins. AB - Isoprenoids are synthesized in all living organisms and are incorporated into diverse classes of end-products that participate in a multitude of cellular processes relating to cell growth, differentiation, cytoskeletal function and vesicle trafficking. In humans, the non-sterol isoprenoids, farnesyl pyrophosphate and geranylgeranyl-pyrophosphate, are synthesized via the mevalonate pathway and are covalently added to members of the small G protein superfamily. Isoprenylated proteins have key roles in membrane attachment and protein functionality, have been shown to have a central role in some cancers and are likely also to be involved in the pathogenesis and progression of atherosclerosis and Alzheimer disease. This review details current knowledge on the biosynthesis of isoprenoids, their incorporation into proteins by the process known as prenylation and the complex regulatory network that controls these proteins. An improved understanding of these processes is likely to lead to the development of novel therapies that will have important implications for human health and disease. PMID- 16378246 TI - Activation of the human FP prostanoid receptor disrupts mitosis progression and generates aneuploidy and polyploidy. AB - Studies have shown prostaglandin F(2alpha) (PGF(2alpha)) to be an endogenous tumor promoter in mouse models of skin carcinogenesis; however, the mechanisms by which PGF(2alpha) affects cell cycle events remain unknown. Here we performed cell cycle analyses on HEK cells stably expressing the human FP receptor and found that treatment with PGF(2alpha) delays mitosis and is associated with an increased expression of cyclin B1 and Cdc2 kinase activity. In addition, multipolar spindles and misaligned chromosomes were observed in a significant proportion of cells treated with PGF(2alpha). Defective cytokinesis was also observed which resulted in gross aneuploidy and polyploidy. Expression of dominant negative Rho attenuated the cell cycle delay and prevented the generation of micronuclei following treatment with PGF(2alpha). This suggests that FP receptor activation of Rho signaling by PGF(2alpha) can interfere with nuclear division. Aneuploidy is associated with genomic instability and may underlie the tumor-promoting properties of PGF(2alpha). PMID- 16378248 TI - Using CQI to address resident concerns about precepting. PMID- 16378249 TI - Quality health care for the underserved. PMID- 16378250 TI - Family physicians and the ethics committee. PMID- 16378251 TI - Thanksgiving thoughts: remembering to appreciate. PMID- 16378252 TI - Care of patients with disabilities: an important and often ignored aspect of family medicine teaching. PMID- 16378253 TI - A sequence of unfortunate events. PMID- 16378254 TI - Communicating with terminal patients: lessons from "wit" and students. PMID- 16378255 TI - Medical students' ability to care for lesbian, gay, bisexual, and transgendered patients. AB - BACKGROUND AND OBJECTIVES: Our objective was to assess medical students' ability to care for lesbian, gay, bisexual, and transgender (LGBT) patients and to identify potential deficiencies in medical school curricula pertaining to this care. METHODS: Between March 1 and April 15, 2004, third- and fourth-year medical students at a metropolitan medical school were sent an e-mail requesting participation in a confidential on-line survey of 64 quantitative questions designed to assess their ability to care for LGBT patients. RESULTS: A total of 248 of 320 (77.5%) students responded. Medical students with greater clinical exposure to LGBT patients reported more frequent sexual history taking with LGBT patients, had more positive attitude scores, and possessed higher knowledge scores than students with little or no clinical exposure. Overall, on the 13-item attitude survey, the mean was 4.15 (5 = most positive, SD = .55, range 1.86 5.00), indicating a desire and willingness to provide health care to LGBT patients. The mean score on the 14-item knowledge test was 60% (SD = .12) correct. CONCLUSIONS: Medical students with increased clinical exposure to LGBT patients tended to perform more comprehensive histories, hold more positive attitudes toward LGBT patients, and possess greater knowledge of LGBT health care concerns than students with little or no clinical exposure. PMID- 16378257 TI - An absent pulse is not sensitive for the early detection of peripheral arterial disease. AB - BACKGROUND: This study's objective was to determine the test characteristics of pedal pulse palpation in the diagnosis of peripheral arterial disease (PAD) when compared to the more widely recommended screening tool, the ankle-brachial index (ABI). METHODS: We screened patients > 50 years of age for PAD within primary care clinics in Houston. PAD was diagnosed by an ABI of <0.9. At each visit, pedal pulse palpation was performed for each leg. Of the patients who screened positive for PAD by ABI, we determined the sensitivity, specificity, and positive predictive value of pulse palpation. RESULTS: We enrolled 403 patients with a mean age of 63.8 +/- .36 years. The prevalence of PAD was 16.6% (67 patients total). Of the 45 patients with disease involving their left leg, 37 (82.2%) had a palpable pulse. Of the 37 patients with disease involving their right leg, 25 (67.6%) had a palpable pulse. The sensitivity of a non-detectable pulse for the diagnosis of PAD was 17.8% and 32.4% for the left leg and the right leg, respectively. The specificity of pulse palpation for the detection of PAD was 98.7% and 97.8% for the left leg and the right leg, respectively. CONCLUSIONS: Pulse palpation is not sensitive for the detection of PAD compared to ABI. More than two thirds of the patients within our cohort with PAD of either the left or right leg had a detectable pulse. PMID- 16378256 TI - Training and evaluating tobacco-specific standardized patient instructors. AB - BACKGROUND AND OBJECTIVES: A comprehensive training program to develop tobacco specific standardized patient instructors (SPIs) was implemented and evaluated at Wake Forest University. METHODS: Descriptive statistics were used to assess SPIs' experience with the training program and medical students' perceptions of the SPI student interaction. Two standardized scales, used to assess student performance on counseling (Tobacco Intervention Risk Factor Interview Scale [TIRFIS]) and cultural competency (Tobacco Beliefs Management Scale-Tobacco Cultural Concerns Scale [TBMS-TCCS]), were tested for internal and interrater reliability and sensitivity to varied student performance. Costs of the program were measured. RESULTS: SPIs highly rated the content, organization, and presenters of the training program. Medical students positively evaluated their experience with the SPIs. The TIRFIS and TBMS-TCCS subscales demonstrated good internal reliability, and inconsistencies in ratings by different SPIs were minimal. In addition, a range of scores on both measures attest to the sensitivity of the instruments to assess variations in student performance. Significant start-up costs are associated with developing this training program, although costs decline when SPIs are retained long term. CONCLUSIONS: The SPI training program was effective in developing a cohort of knowledgeable and reliable SPIs to train medical students in ways to improve their tobacco intervention counseling skills. Retaining SPIs long term should be a primary goal of implementing a cost effective, successful training program. PMID- 16378258 TI - The Morehouse Faculty Development Program: evolving methods and 10-year outcomes. AB - BACKGROUND AND OBJECTIVES: African American physicians remain underrepresented among all medical school faculty, including faculty in departments of family medicine. This paper reports on a faculty development effort aimed at increasing the number and academic skills of underrepresented minority faculty. METHODS: In 1992, Morehouse School of Medicine began a faculty development program. The program trains faculty and community-based preceptors in teaching, scientific writing, grant writing, research, and minority career issues. Formats now include a 1-year longitudinal program, 4-6-week stand-alone modules, and an executive faculty development program for physicians from across the nation. Evaluation measures include participant enrollment, completion rate, participant feedback, and self-reported academic competencies before and after the program. RESULTS: A total of 113 participants completed the program from 1992-2003. Only seven enrollees failed to complete the program. Of 113 graduates, 104 (92.0%) were ethnically African American, Afro Caribbean, or African, while only two were white, non-Hispanic. More than four out of five (81%) now spend at least some time teaching on a regular basis, and 71% spend more than 25% time in teaching roles. Self-reported before-after competencies in specific academic skills such as teaching, writing, research, and grant writing rose from 2.7 to 4.1 on a 5 point scale. CONCLUSIONS: Faculty development is a potentially effective strategy for increasing diversity in academic primary care. Historically black and Hispanicserving institutions can make contributions to training minority faculty. More-rigorous study could elucidate which program elements have the greatest effect on minority faculty academic career choice, scholarly productivity, and career trajectory and the extent to which these programs could be adapted to majority institutions. PMID- 16378259 TI - Development of a practice-based patient cohort for primary care research. AB - BACKGROUND AND OBJECTIVES: Efforts to develop "laboratories" for primary care research have largely focused on the development of networks that recruit subjects on a project-specific basis. We sought to develop an alternative model- a representative cohort of adult primary care patients maintained for use in multiple projects. METHODS: In 2001, research assistants in waiting areas of a representative sample of 16 family medicine practices in North Carolina approached all adult patients during a 4-week period. Follow-up has been maintained for 3 years. In 2004 and 2005, the cohort was refreshed by adding eight new practices. Each consenting subject was administered a four-page self report questionnaire that included items on demographics, risk factors, health status, and quality of life. RESULTS: Of 10,649 eligible patients approached in 2001, 6,811 (64%) completed the enrollment questionnaire, of whom 5,575 (81.9%) consented to be included in the cohort. African Americans, Latinos, and older persons were enrolled at rates paralleling the state's adult population. Poor general health, chronic illness, and risk factors for chronic disease were more prevalent in the cohort than in the general population. Over 3 years, cohort members were included in multiple studies, and 77% of the original cohort remained active. The per-subject enrollment cost varied between 27 US dollars and 45 US dollars; annual program maintenance costs were estimated at approximately 35,000 US dollars. CONCLUSIONS: The research cohort has these advantages over traditional practice-based research networks: patient focus rather than physician focus, a structure that places few demands on practices, ability to target racial and ethnic minorities, and a better-defined patient population. As is true of all aspects of research infrastructure, the cost of development and maintenance is significant. PMID- 16378263 TI - Africa: some thoughts on the medical brain drain. PMID- 16378264 TI - Saudi Arabia: a proposal for the future of family medicine. PMID- 16378265 TI - Australia: the N of 1 trial, an underappreciated research method. PMID- 16378266 TI - Hepatitis B seroprevalance and risk factors in urban areas of Malatya. AB - BACKGROUND/AIMS: To determine the prevalence of hepatitis B viral markers and to assess possible risk factors in urban areas of Malatya. METHODS: This was a sero epidemiological, community based cross-sectional study and included 646 participants( female 352, male:294) from 192 houses. A face to face questionnaire was carried out and HBsAg, anti-HBc and anti- HBs markers were analyzed from blood samples using Micro ELISA technique. RESULTS: The prevalence of HBsAg, anti HBc and anti-HBs were found to be 6.0%, 29.3% and 30.3% respectively. In the final logistic regression, HBV infection (=anti HBc+) was independently associated with the age group of 21 years and older (OR=3.7, 95% CI=1.884-7.494), in illiterate subjects (OR=2.1, 95% CI=1.180-3.326), in farmers and labourers (OR=2.8, 95% CI=1.042-7.953) and in these with multiple sexual partners (OR=2.1, 95% CI=1.574-8.168). In addition, HBV infection was significantly higher in circumcised male children compare to uncircumcised ones ( chi2=5.58, P=0.01), in ones who gave birth to child at home compare to in ones who gave birth to a child at hospital ( chi2=13.86, P=0.0001). CONCLUSION: The results of our study indicate that Malatya province has a moderate endemicity with regard to HBV infection. PMID- 16378267 TI - The effect of recombinant hepatitis B vaccine therapy in chronic hepatitis B infection. AB - BACKGROUND/AIMS: Immune-modulator and antiviral treatments for carriers of hepatitis B virus are known to have poor efficacy with a high cost and frequent side effects, which has led to investigation of new treatment modalities. The aim of this study was to determine the efficacy of recombinant hepatitis B vaccine in the treatment of patients with chronic hepatitis B (HBV DNA >5 pg/ml, ALT>60) infection diagnosed histopathologically and asymptomatic carriers (HBV DNA<5 pg/ml, ALT <40, Anti Hbe positive) of the virus. METHODS: The vaccine (Gen Hevac B Pasteur) was administered at baseline and at months one and six to patients with chronic hepatitis and asymptomatic carriers. Ten cases with chronic hepatitis B infection were assigned to a control group to whom no treatment was given. Biochemical and microbiological investigations were performed at baseline and at months three, six and twelve in all cases. Seroconversion of Hbe Ag, loss of HBV DNA and normalization of ALT were considered as a positive response. RESULTS: Patients with chronic hepatitis B who were given the vaccine were found to have significantly low levels of HBV DNA at 12 months (63.2+/-20pg/ml) compared to baseline values (174.4+/-36.9pg/ml), while controls were found to have high levels of HBV DNA at 12 months (223.1+/-33pg/ml) compared to baseline values (165.2+/-33.2pg/ml) (p<0.05). At 12 months, HBV DNA had become negative in seven of 19 patients given the vaccine (36.8%) Four patients with chronic hepatitis (36.35%) were observed to have HBeAg seroconversion and one patient (5.2%) HBsAg seroconversion at the end of 12 months and there were four (21.05%) patients who responded positively to vaccine therapy in this group. Asymptomatic carriers and controls did not have seroconversion of HBs Ag. Also, HBV DNA did not become negative in controls. CONCLUSION: It may be concluded that recombinant hepatitis B vaccine is effective in the treatment of chronic hepatitis B. PMID- 16378268 TI - Diagnosis and surgical treatment of complicated liver echinococcosis. AB - BACKGROUND/AIMS: Complicated liver echinococcosis considerably worsens the prognosis of the disease. In this study the treatment of patients with complicated disease was analyzed. METHODS: The records of 136 patients who had undergone surgery in our clinic during the last decade were evaluated. RESULTS: Among all complications, the most common were lesions of the bile ducts (88 patients) and suppuration of the cysts (77 patients). Twenty-nine cases had a combination of two complications. Surgery was based on the nature of complications, number and sizes of hydatid cysts and patients general condition. The surgical treatment of choice was echinococcectomy, with possible liquidation of the fibrous capsule cavity. In cases with complications such as suppuration, remaining bile fistulas, large cyst cavities or extrahepatic location, external drainage is obligatory. Post-operative complications occurred in 43 patients (31.6 %), 22.4 % of them being specific in nature. Postoperative mortality was 2.2 % (three patients). CONCLUSION: Timely diagnosis, appropriate investigation and well planned surgical techniques may improve the outcome of complicated liver echinococcosis. PMID- 16378269 TI - The effect of recombinant growth hormone on intestinal anastomotic wound healing in rats with obstructive jaundice. AB - BACKGROUND/AIMS: Several clinical and experimental studies have shown that obstructive jaundice delays wound healing. Growth hormone may prevent delayed wound healing, since it has effects on the release of mediators in jaundice, as well as increasing the protein synthesis. METHODS: Forty male Wistar rats were allocated to four groups: Group I (n=10): intestinal anastomosis to normal small bowel, Group II (n=10): intestinal anastomosis to normal small bowel followed by growth hormone therapy (2mg/kg/day, subcutaneously), Group III (n=10): intestinal anastomosis to obstructive jaundice rat's small bowel, Group IV (n=10): intestinal anastomosis to obstructive jaundice rat's small bowel followed by growth hormone therapy at the same dosage The animals were observed for seven days then killed. Intraabdominal adhesions, anastomotic complications and anastomotic bursting pressures were recorded and tissue samples from the anastomotic site were obtained to measure hydroxyproline levels and for histopathologic examination. RESULTS: Growth hormone had a beneficial effect on the healing of intestinal anastomosis in both jaundiced and non-jaundiced rats. This was demonstrated by clinical and mechanical parameters such as a significant increase in anastomotic bursting pressure, hydroxyproline content and histopathological scores. CONCLUSION: Growth hormone reverses the adverse effects of obstructive jaundice on small bowel anastomotic healing. It can be hypothesized that this effect is due to augmentation of insulin-like growth factors, protection of hepatocytes, enhancement of intestinal epithelization, and reversal of the resultant malnutritional state caused by growth hormone in obstructive jaundice. PMID- 16378270 TI - Hypersensitive esophagus: can it be classified as a subgroup of gastroesophageal reflux disease? AB - BACKGROUND/AIMS: Gastroesophageal reflux disease is one of the most common gastrointestinal diseases of adulthood. Hypersensitive esophagus is a new term which defines patients with reflux symptoms, negative endoscopic findings and normal acid contact time on 24 hour pH analysis, although there is a convincing relationship between symptoms and acid reflux episodes on pH analysis- positive symptom index. In this study, the frequency of hypersensitive esophagus among patients with heartburn was determined and demographic findings, symptoms, manometric and pHmetric findings were compared among hypersensitive esophagus, non-erosive reflux disease, erosive reflux disease and normals. METHODS: Patients admitted to hospital with heartburn and without any cardiac, severe gastric or duodenal pathologies (except minimal antral gastritis) and with no abnormal manometric findings suggestive of esophageal motility disorders were included in the study. All patients were questioned about age, height, weight, educational status and intestinal and extraintestinal reflux related symptoms. Upper endoscopy, esophageal manometry and 24 hour pH monitoring were performed respectively. RESULTS: Of the 44 patients (17 male, 27 female) included in the study, seven (16%) had hypersensitive esophagus, 15 (34%) had non-erosive reflux disease, seven (16%) had erosive reflux disease and 15 (34%) had normal findings. The female ratio in the hypersensitive esophagus and normal groups was higher while the male ratio was higher in erosive reflux disease. There was no significant difference among groups according to age, body mass index and symptoms. There was also no significant difference according to manometric findings. De Meester scores were significantly lower in hypersensitive esophagus and normal groups while symptom index was highest in the hypersensitive esophagus group. Reflux was observed especially in the upright position in all groups. CONCLUSION: The ratio of hypersensitive esophagus, gastroesophageal reflux disease and no pathology in patients with heartburn was 16%, 50% and 34% respectively. It would be appropriate to accept hypersensitive esophagus as a subgroup of reflux disease as it has both symptoms and some manometric and pHmetric changes. PMID- 16378271 TI - Upper gastrointestinal findings in oral lichen planus. AB - BACKGROUND/AIMS: Lichen planus is an inflammatory disease of the skin and mucous membranes. Oral mucosa is known to be frequently affected by the disease, but it has also been observed that gastrointestinal mucosas are sometimes involved. METHODS: In this study, the upper gastrointestinal tract was investigated endoscopically and histopathologically in 20 patients with oral lichen planus. RESULTS: Endoscopy determined antral gastritis in seven patients, esophagitis in four, bulbitis in three, chronic duodenal ulcer in one and esophageal sphincter dysfunction in one patient. Lichen planus-like changes on the esophageal mucosa at histopathological examination were found in one patient. Findings in the other patients were as follows: chronic atrophic gastritis (nine), helicobacter pylori infection (nine), esophagitis (two), bulbitis (two) and erosive gastritis (one). CONCLUSION: Patients with Lichen planus should be evaluated for possible gastro intestinal involvement with endoscop. PMID- 16378272 TI - A reference interval study of urinary lactulose excretion: a useful test of intestinal permeability in adults. AB - BACKGROUND/AIMS: Measurement of lactulose excretion in urine following oral ingestion is used as a noninvasive method of assessing small intestinal permeability. The aim of this study was to determine the adult reference interval of urinary lactulose excretion following oral administration. METHODS: Thirty seven volunteers (mean age 37.3 +/- 15 years) with no history of gastrointestinal disease were recruited as study subjects. All subjects were fasted overnight (10 hours), emptied their bladders and then drank a 50-mL solution containing 15 mL of Duphalac (10 g lactulose) and 35 mL flavoured water. Urine was collected for six hours in a bottle. Comparisons were made for 37 samples. RESULTS: In adults, the mean urinary lactulose concentration was 0.58 +/- 0.39 mmol/L. CONCLUSION: This study determined the adult reference range for lactulose excreted in the urine, using a sensitive quantitative assay based on hydrolysing lactulose and enzymatically assaying fructose, one of the component monosaccharides. PMID- 16378273 TI - Effects of alpha tocopherol and verapamil on liver and small bowel following mesenteric ischemia-reperfusion. AB - BACKGROUND/AIMS: We have previously shown that alpha tocopherol is a potent antioxidant which prevents reperfusion injury in a kidney and small bowel autotransplant model. In this study, the effect of systemic alpha tocopherol and verapamil on small bowel and hepatic functions following mesenteric ischemi reperfusion was evaluated. METHODS: Fourty male Wistar Albino rats (weight, 250 300 g) all subjected to an ischemia-reperfusion experiment were divided into four groups of 10 as follows: Group 1: (SHAM), Group 2, given prophylactic and systemic alpha tocopherol, Group 3: given verapamil and Group 4: given (both verapamil and alpha tocopherol). RESULTS: Glutaminase activities 120 minutes after reperfusion were found to be significant in liver tissues (p=0.004). The highest to the lowest glutaminase activities in liver tissue at 120 minutes after reperfusion were in Group 1, Group 3, Group 2 and Group 4 respectively. Significant differences in MDA levels were found in the small bowel at 30 minutes and 120 minutes time points (p<0.05). There were statistically significant higher glutaminase levels at 30 minutes and 120 minutes of reperfusion in the small bowel, especially in Group 4 (p=0.005). CONCLUSION: Both small bowel and liver injuries reperfusion, can be decreased by prophylactic use of alpha tocopherol and verapamil. Glutaminase activity in liver tissue can also be affected by small bowel ischemia-reperfusion. PMID- 16378274 TI - Abdominal surgical emergency in the elderly. AB - BACKGROUND/AIMS: Longer life expectancy has created an increasing demand for surgical care of the elderly. Abdominal surgical emergencies are potentially serious and life-threatening conditions for this group of patients. The aim of this study was to evaluate the records of elderly patients undergoing emergency surgical treatment. METHODS: A total of 181 patients aged 60 years and above who had undergone emergency surgical interventions were retrospectively analysed according to demographic features, indications for emergency surgery, postoperative clinical course and outcome. RESULTS: Sixty-one percent of the patients were male, with an average age of 70.3 (range 60-95) years. Octogenarians constituted 19% of our series. Causes of surgical emergency were acute cholecystitis (31.5%), strangulated hernia (18.2%), hollow viscus perforation (17.1%), bowel obstruction (10.5%), acute mesenteric ischaemia (9.4%), acute appendicitis (8.3%) and upper gastrointestinal haemorrhage (5%). Gallstones had been previously detected by ultrasound in 25 (45.5%) of 55 patients with acute calculous cholecystitis. Thirty (93.8%) of 32 patients were aware of their external hernias prior to incarceration. Twenty seven patients (14.4%) died in the early postoperative period, with acute mesenteric vascular occlusion being the leading cause of death: A higher mortality rate was noted in mesenteric ischaemia (76.5%), gastrointestinal bleeding (22.2%) and in perforation (19.4%) cases, being 36.8% (21/57) in patients with these three severe conditions and 4.8% (6/124) in the remaining patients. CONCLUSIONS: Acute calculous cholecystitis and external strangulated hernias, which are generally preventable, were the most common surgical emergencies. Surgical treatment of acute mesenteric ischaemia, gastrointestinal haemorrhage and peritonitis secondary to free perforation had a worse prognosis in older patients. PMID- 16378275 TI - The diagnostic value of ultrasonography-guided fine needle aspiration biopsy from liver and pancreas. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the results of ultrasonographic guided fine needle aspiration biopsy conducted at our clinic between 1999 and 2000. METHODS: Biopsy was taken from either the liver or pancreas of 40 patients (29 male, 21 female) with a mean age of 59.7+/-11.99 years. RESULTS: The diameter of lesions was 2-15 cm, with 26 of 33 lesions in the liver being malignant. There was a solitary lesion in the liver being malignant. There was a solitary lesion in the pancreas in seven cases and these were found to be adenocarcinomas. CONCLUSIONS: This procedure is concluded to be a highly accurate diagnostic method, which is both fast and practical in clinical practice. PMID- 16378276 TI - McArdle's disease: case report and review of the literature. AB - McArdle's disease, or myophosphorylase deficiency, is one of the most common muscle glycogenoses and typically presents in childhood or adolescence with exercise intolerance, myalgia, myoglobinuria, and cramps in the exercising muscle. We report the case of a 21-year-old male patient with liver enzyme elevation who had a history of weakness, exercise intolerance, and muscle cramps since childhood. His sister (a three-year- old) suffered from similar symptoms. Laboratory results showed that serum creatinine phosphokinase levels were elevated serum lactate did not rise on ischemic exercise testing, while muscle biopsy showed subsarcolemmal and intermyofibrillar periodic acid schiff-positive vacuoles filled with glycogen. This case report underlines the importance of taking into account rare metabolic diseases such as muscle glycogenoses in the evaluation of patients with elevated liver enzymes. PMID- 16378277 TI - Alanine aminotransferase deficiency in a hepatitis B surface antigen positive patient presenting with acute hepatitis. AB - This report presents a hepatitis B surface antigen positive case presenting with acute hepatitis and with findings of low serum alanine aminotransferase in contrast to very high levels of aspartate aminotransferase. A 64 year-old female patient was admitted to our hospital with fatigue and jaundice. Hepatitis B surface antigen was positive. During follow up, aspartate aminotransferase levels remained very high, while alanine aminotransferase levels continued to be extremely low. Additionally, all of the patients five daughters had low alanine aminotransferase levels. The clinical importance of alanine aminotransferase deficiency is still unclear. PMID- 16378278 TI - The role of surgery in the treatment of instussusception in celiac disease. AB - Small bowel intussusceptions comprise fewer than 10% of all pediatric patients with intussusceptions and most of them are secondary to another pathology. In this report, we discuss the role of surgery in the treatment of intussusception in celiac disease. A 13-month-old girl was admitted with a three day history of progressive abdominal distention and vomiting of bile. There were air-fluid levels on supine abdominal X-ray and ultrasonographic examination demonstrated an intussusception. At surgery, two separate small bowel intussusceptions were encountered. The postoperative course was uneventful. Due to a history of frequent diarrhea and mild abdominal distention developing after the age of seven months, further studies for celiac disease were initiated. Antigliadin and antiendomysium antibodies were found to be strongly positive. Celiac disease was also confirmed by endoscopic small bowel biopsy. Children who present with chronic or transient intestinal obstruction should also be evaluated for underlying celiac disease. Nevertheless, the surgical decision should be based upon clinical observation in this group of patients. PMID- 16378279 TI - Crohn's colitis with extraintestinal manifestations: a case report. AB - The etiopathogenesis of inflammatory bowel disease remains unclear, with various extraintestinal features often found with the disease. Sometimes more than one extraintestinal finding is found in the same patient. In this case report, a patient with more than one extraintestinal manifestation of Crohn's disease is presented. Extensive thrombi in the venous system and pyoderma gangrenosum in the left lower extremity was observed but there was complete recovery following treatment with both systemic and topical agents. PMID- 16378280 TI - Obesity and related parameters of non-alcoholic steatohepatitis. AB - BACKGROUND/AIMS: The aim of this prospective study was to evaluate the clinical, biochemical and histopathological parameters of nonalcoholic steatohepatitis and the conditions associated with this disease. METHODS: Twenty-four patients were included in the study, each having been diagnosed with nonalcoholic steatohepatitis on the basis of liver biopsy and elimination of other possible causes of elevated aminotransferase levels. Measurements of degree of obesity, liver enzymes and serum lipids were recorded before liver biopsy and reevaluated after one or two months of a standard exercise and diet program. Serum insulin levels were also measured. Each liver biopsy was histologically examined for steatosis, inflammation, fibrosis, necrosis and iron storage, and semiquantitave assessment of these was recorded for three separate hepatic zones. RESULTS: The prevalence of obesity in the group was 79.2%, while the figure for overt and latent diabetes mellitus combined was 33.3%, and for hyperlipidemia was 83.3%. Compared to the rest of the group, the patients with severe steatosis had significantly higher serum lipid (particularly high triglyceride) and insulin levels (p<0.05 for both). There was a correlation between steatosis and obesity (p=0.06). More severe obesity, higher insulin and elevated aspartate aminotransferase were positively correlated with inflammation, whereas elevated serum triglyceride was negatively correlated with inflammation. There was a tendency towards normalization of liver enzyme levels after weight loss and dietary restrictions. CONCLUSIONS: Obesity and hyperlipidemia were associated with nonalcoholic steatohepatitis in the group studied. Obesity is not a factor in every case, but the study showed that restricted diet and exercise are significant forms of therapy for nonalcoholic steatohepatitis. PMID- 16378281 TI - The value of fine needle aspiration biopsy in the diagnosis of metastatic liver tumours. AB - BACKGROUND/AIMS: Fine needle aspiration biopsy is a useful tool in the diagnosis of primary malignancies and metastatic lesions of the liver. The aim of this study was to determine the types and features of tumors diagnosed by this method and the difficulties in differential diagnosis. METHODS: Fine needle aspiration biopsy smears from 704 patients with metastatic liver lesions were reviewed. RESULTS: Among the metastastatic carcinomas in which their primary origin was identified, pulmonary carcinomas were the largest group. While colon adenocarcinoma was most prevalent (21.65%) where the primary origin of metastatic tumors was identified, followed by breast carcinoma (20.10%) and gastric adenocarcinoma (19.59%). The cases which cannot be differentiated from hepatocellular carcinoma in cytologic examination are invasive ductal carcinoma, renal cell carcinoma and squamous cell carcinoma. CONCLUSIONS: Pulmonary and colon carcinomas are the common metastatic tumors of the liver. PMID- 16378283 TI - Antiphospholipid antibodies and lipoprotein (a) as etiologic or contributory factors in patients with idiopathic cavernous transformation of portal vein. AB - BACKGROUND/AIMS: Antiphospholipid antibodies consisting of anticardiolipin antibodies and lupus anticoagulant are strongly associated with thrombosis in adult patients. It is also well known that there is a close relationship between antiphospholipid antibodies and lipoprotein (a) in thrombous formation. The aim of this study was to determine whether antiphospholipid antibodies and lipoprotein (a) have any effect on the formation of thrombosis in the portal vein of patients with 'idiopathic' cavernous transformation of the portal vein. METHODS: Twenty seven patients with idiopathic cavernous transformation of the portal vein (Group 1) seen at Hacettepe University Hospital were identified and prospectively studied. All were investigated for antiphospholipid antibodies and lipoprotein (a). Anticardiolipin antibodies, lupus anticoagulant and lipoprotein (a) were measured using commercially available kits. The findings of these 27 patients were compared with those of 20 healthy control subjects (Group 2). RESULTS: Anticardiolipin antibodies, especially ACA Ig G and lipoprotein (a) levels were found to be higher than of healthy controls and statistically significant differences were documented in two of these parameters, which seems to play an important role in thrombous formation in the portal vein. There was no correlation for lupus anticoagulant between the two groups. CONCLUSIONS: Anticardiolipin antibodies and lipoprotein (a) are strongly associated with thrombosis in the portal vein, producing a favorable medium for and acting as contributory factors in thrombous formation. It is suggested that these factors should be evaluated carefully in patients with 'idiopathic cavernous transformation of the portal vein'. PMID- 16378282 TI - Percutaneous transhepatic venoplasty: an alternative treatment for Budd-Chiari syndrome. AB - BACKGROUND/AIMS: In patients with Budd-Chiari syndrome due to short segment hepatic vein stenosis where percutaenous transluminal venoplasty is not successful, percutaneous transhepatic balloon venoplasty may be a valid treatment option. The aim of this prospective study was to evaluate the effects of this procedure for the treatment of patients with Budd-Chiari syndrome, in whom transluminal cannulation was unsuccessful. METHODS: Ten patients with short segment occlusion of the hepatic veins were treated by percutaneous transhepatic balloon venoplasty between January 1997 and January 2000. The median follow-up period of these patients was 20 months (2-33 months). RESULTS: The procedure was unsuccessful in two patients. Eight patients (five men, three women) with a median age of 28 (range, 15-61) years were treated by percutaneous transhepatic balloon venoplasty and in seven of them, clinical symptoms including abdominal distension and ascites, resolved completely. Long term anticoagulation therapy was not given to the patients. One patient with advanced stage liver disease died of variceal bleeding two months after the procedure. During follow-up, symptomatic reocclusion requiring dilatation occurred in three patients. CONCLUSIONS: Percutaneous transhepatic balloon venoplasty is an alternative treatment option for selected patients with Budd-Chiari syndrome when transluminal cannulation of the hepatic veins is not possible. Long term anticoagulation therapy seems to be necessary in these patients. PMID- 16378284 TI - Detection of Helicobacter pylori using nested polymerase chain reaction and rapid urease test in gastric biopsy samples. AB - BACKGROUND/AIMS: The purpose of this study was to compare nested polymerase chain reaction and rapid urease test findings in the diagnosis of Helicobacter pylori infection. METHODS: Two gastric biopsy specimens were obtained from each of 64 patients and polymerase chain reaction and rapid urease test were performed. DNA extraction was followed by amplification with two primer pairs from the urease A gene of Helicobacter pylori genome. RESULTS: Fourty two patients (65.6%) had a positive rapid urease test result while 22 (34.4%) had a negative result for Helicobacter pylori. Sixty of 64 patients (93.8 %) had a positive result with the nested polymerase chain reaction method. Four patients with negative nested polymerase chain reaction results also had negative rapid urease test results. All of the eighteen patients with rapid urease test-negative results were positive with nested polymerase chain reaction. The number of nested polymerase chain reaction-positive patients (93.8%) was significantly higher than rapid urease test-positive (65.6%) and first round polymerase chain reaction-positive (53.1%) patients (P<0.001). CONCLUSIONS: Our results indicate that the nested polymerase chain reaction is more specific and sensitive than the rapid urease test for detecting Helicobacter pylori in gastric biopsy samples. PMID- 16378285 TI - The relationship between Helicobacter pylori intensity and histopathological findings in cases with chronic gastritis and duodenal ulcer. AB - BACKGROUND/AIMS: Helicobacter pylori infection affects many people in developing countries. The inflammation it induces can cause malignant gastric lymphoma and also gastric carcinoma, depending on the intensity of inflammatory reaction, specific properties related to the strain and the host response. METHODS: Fifty patients (35 cases with gastritis and 15 with duodenal ulcer) were included in the study. Hematoxylin-eosin paraffin sections were prepared from their endoscopic biopsies and Helicobacter pylori presence, activity of the infection, lymphoplasmacytoid cell infiltration and the degree of atrophy were determined semiquantitatively. Toludin O stain was applied to determine Helicobacter pylori intensity. RESULTS: We found a statistically significant positive correlation of Helicobacter pylori intensity and activity of infection, lymphoplasmacytoid cell infiltration and atrophy in the 35 cases with gastritis and the relationship between Helicobacter pylori intensity and inflammatory activity was statistically significant in the 15 cases with duodenal ulcer. CONCLUSION: As morphological characteristics are crucial for the early diagnosis and treatment of gastric malignities; histopathological confirmation of helicobacter intensity is important. PMID- 16378287 TI - The effect of methylene blue on peritoneal adhesion formation. AB - BACKGROUND/AIMS: Postoperative adhesions following abdominopelvic surgery may cause significant morbidity. In this study, the effect of different doses of methylene blue in the formation of adhesions and the role of allopurinol in revealing the possible mechanism of action was evaluated. METHODS: Seventy five male Wistar-Albino rats weighing 200g-220g. were divided into five groups of 15 rats each. After laparotomy, cecal serosal abrasions were performed. In Group I, 1ml of saline was administered intraperitoneally, while Group II, III and IV were given 1mg/kg, 5mg/kg and 9mg/kg of intraperitoneal methylene blue respectively. In Group V after 1 ml of saline administration at surgery, allopurinol 30 mg/kg/ day was administered postoperatively for 14 days. On day 14, laparotomies were repeated. Adhesions were graded and tissue samples were taken from incisions and adhesions. Hydroxyproline contents representing adhesions were measured quantitatively. RESULTS: Low dose methylene blue (Group II) caused inhibition of adhesion formation when compared with that of other groups (p<0.05). With a higher dose (Group III), this effect disappeared and methylene blue actually induced adhesion formation (p<0.05) at the highest dose (Group IV). Allopurinol treatment (Group V) also provoked adhesion formation when compared with Group I (p<0.05). CONCLUSIONS: Methylene blue has diverse effects on postoperative adhesions in a dose-dependent manner. While with low doses there is evidence of inhibition, it promotes adhesion formation at higher doses. As allopurinol also provokes adhesion formation, it is thought that the preventive role of methylene blue may not be due to its free oxygen radical inhibitor effect but the mechanism has yet to be clarified. PMID- 16378286 TI - The prevalence of manifest and latent celiac disease in type 1 diabetes mellitus. AB - BACKGROUND/AIMS: Celiac disease and type 1 diabetes mellitus are both autoimmune diseases which have a common genetic predisposition. The aim of this study was to determine the prevalence of manifest and latent celiac disease in type 1 diabetic patients. METHODS: Anti-endomysium IgA was tested by indirect immunofluorescence using sections of human umbilical cord for screening in 100 adult patients with type 1 diabetes mellitus and in 80 age and sex matched controls with no known disease. Distal duodenal biopsy, human leukocyte antigen typing, urinary D-xylose excretion test, stool analysis, biochemistry profile, blood counts, serum ferritin level and small intestinal radiography were performed in anti-endomysium IgA positive cases. Small bowel biopsy specimens consistent with celiac disease were defined as manifest celiac disease, while positive antiendomysium IgA and normal intestinal histology with the presence of human leukocyte antigen class II antigens consistent with the disease were defined as latent celiac disease. RESULTS: Anti-endomysium IgA was positive in eight diabetic patients, while it was negative in all controls. Celiac disease was found in a total of six (6%) patients, four with manifest and two with latent disease. Only one patient had symptoms. CONCLUSIONS: The prevalence of celiac disease is increased in patients with type 1 diabetes mellitus. Since many patients may be asymptomatic, it is suggested that all diabetic patients should be screened for this disease. PMID- 16378288 TI - Alveolar hydatid cyst with lung metastasis: a case report. AB - Alveolar hydatid disease is a serious and often fatal condition caused by infection with the metacestode form of echinococcus multilocularis. Liver involvement is seen in 90% of the cases. The parasite develops within the liver as a rapid invasive pseudomalignant growth and may make distant metastasis. We present the case of a 37 year old woman admitted to our gastroenterology department with obstructive jaundice and then diagnosed as alveolar hydatid cyst with lung metastasis. This case is presented due to its rare prevalence. PMID- 16378289 TI - Ogilvie's syndrome in a patient with multiple myeloma. AB - Ogilvie's syndrome is characterized by physical examination and radiologic findings indicative of mechanical obstruction but in which no physical obstructive process can be found. Many factors have been associated with this syndrome which include electrolyte imbalance, systemic infection, drugs, and occasionally, neurologic disease. A case of acute colonic pseudoobstruction is presented which developed in a patient with multiple myeloma. The patient presented with severe thoracic pain, persistent and increasing abdominal distention and lack of bowel sounds. Plain radiography and ultrasonography revealed massive dilatation of the right and transverse colon. Nasogastric aspiration was initiated and all analgesic drugs were withdrawn. Erythromycin was given for nine days as prokinetic and a rectal tube was inserted for one day. Abdominal distention gradually disappeared within one day of nasogastric and rectal tube insertion and with multiple myeloma management. Ogilvie's syndrome is a very rare complication of multiple myeloma. Only one case of Ogilvie's syndrome with multiple myeloma has been reported in the literature. This case report of Ogilvie's syndrome in a patient with multiple myeloma is the second case report in the literature. PMID- 16378290 TI - Hypereosinophilic syndrome: case report. AB - Hypereosinophilic syndrome is a spectrum of disorders characterized by marked eosinophilia of no identifiable cause and by organ dysfunction. In 1999, a patient with marked weight loss, eosinophilia and a hypodense lesion in the right lobe of the liver was diagnosed as hypereosinophilic syndrome by fine needle aspiration biopsy and bone marrow aspiration biopsy. Echocardiography and thoracal HRCT was found to be normal. The patient was prescribed 30 mg prednisolone daily with the dose being tapered gradually. At the end of treatment, computerized tomography was normal but ultrasonography showed a 1mm x 3mm lesion. Liver scintigraphy was then performed in order to determine the activity of the lesion and was found to be normal. The lesion was therefore thought to be from a previous infection. PMID- 16378291 TI - A rare cause of acute abdominal pain: primary torsion of omentum majus. AB - Primary torsion of the omentum majus is a rare cause of acute abdomen and it commonly mimics acute appendicitis. An eight year old boy was admitted to our clinic with symptoms and clinical findings of perforated appendicitis. The patient underwent emergency laparotomy at which a normal appendix and serosanguinous fluid in the peritoneal cavity were observed. The operative diagnosis was primary torsioned omentum. In the present study, preoperative and operative findings of primary omental torsion that differentiated it from acute appendicitis are discussed in the light of the literature. PMID- 16378292 TI - The frequency of sacroileitis and ankylosing spondylitis in inflammatory bowel disease and HLA-B27 association. AB - BACKGROUND/AIMS: The aim of this study was to investigate the frequency of sacroileitis and ankylosing spondylitis in inflammatory bowel disease and the relationship between disease activity, joint symptoms and HLA-B27 antigen positivity. METHODS: Sacroiliac joint radiography and three phase bone scanning of 33 ulcerative colitis patients (17 active and 16 in remission) and 29 Crohn's disease patients (15 active and 14 in remission) was performed. HLA-B27 was determined in all patients and 60 control subjects. RESULTS: Sacroileitis was found in 10 out of 33 patients with ulcerative colitis (30. 30%) and seven out of 29 patients with Crohn's disease (24. 13%). Of these patients, eleven had active (17.73%) disease and six were in remission (9. 68%). The diagnosis of sacroileitis was made by bone radiography in seven patients (41.18%) and by bone scanning in the other 10 patients (58.82%). A diagnosis of ankylosing spondylitis was made in three patients (17. 64%) who had previously been diagnosed by bone radiography to have sacroileitis. HLA-B27 was positive in six patients (9.67%) with inflammatory bowel disease and three subjects (5%) of the control group. There were no significant differences between these two groups (p>0.05). Compared to the control group, all three patients with ankylosing spondylitis were HLA-B27 positive, the difference being significant (p>0. 01). CONCLUSIONS: The clinical course of sacroileitis is independent of the activitiy of inflammatory bowel disease. Accordingly, patients with inflammatory bowel disease, especially those with sacroileitis, should be investigated for the presence of anklylosing spondylitis. PMID- 16378293 TI - Efficacy of cyclosporin in severe ulcerative colitis attack. AB - BACKGROUND/AIMS: Cyclosporin-A is used as a alternative medical therapy in steroid resistant ulcerative colitis with severe activity. In spite of its known efficacy, the long term effects of are not entirely clear. METHODS: The records of 13 steroid resistant patients treated with cyclosporin-A were retrospectively assessed. Cyclosporin-A had been prescribed orally at a dose of 8 mg/kg/day in four patients and intravenously, 4mg/kg/day in nine patients. Intravenous therapy was changed to oral therapy after one week and patients also received 5-ASA and azathioprine. Steroid treatment was tapered. RESULTS: Ten patients responded to treatment in a mean of nine days (range: 2-30 days). Three patients who did not respond underwent total colectomy on day seven, 11 and 19 of therapy. The 10 patients who initially responded received the drug for an average of 4.9 months; four of these relapsed during and one relapsed soon after discontinuation of therapy. Four of the five patients who relapsed underwent colectomy and the one patient who did not accept surgical intervention continued medical therapy. The remaining five patients (38% of the total group; 50% of the patients who initially responded) remained in remission at the end of an average 17 month follow up period. CONCLUSIONS: Cyclosporin-A therapy in severe ulcerative colitis that is resistant to steroids, provides initial remission in 80% of patients and allows 40% to retain their colon for one year. PMID- 16378294 TI - The course of colonic disease in ulcerative colitis patients with primary sclerosing cholangitis. AB - BACKGROUND/AIMS: Although there are many studies reporting that colonic dysplasia and cancer develop more frequently in ulcerative colitis patients with ulcerative colitis with primary sclerosing cholangitis, there are insufficient data on the course of the colonic disease. In this study, the course of the colonic disease in ulcerative colitis patients with primary sclerosing cholangitis was investigated. METHODS: Data obtained from ten patients with total colitis and accompanying primary sclerosing cholangitis (three females, seven males, mean age: 44.5+/-10.0 years) were compared with data obtained from 64 patients with pancolitis but without primary sclerosing cholangitis (27 females, 37 males; mean age: 42.3+/-17.1 years). RESULTS: The follow-up period was 6.4+/-6.2 years in patients without primary sclerosing cholangitis, 12.7+/-6.2 years in total and 5.1+/-4.0 years (after development of the condition) in patients with primary sclerosing cholangitis (p<0.01). The number of disease attacks (3.7 attacks/yr vs. 0.5 attacks/yr), duration of the active disease (12.9+/-8.0 months vs. 0,3+/ 1.0 months), the number of patients in whom corticosteroids were used (47 patients vs. one patient), the number of patients hospitalized (50 patients vs. one patient) and duration of hospitalization (1.2+/-0.8 months vs. 0,1+/-03 months) were higher in patients with than without primary sclerosing cholangitis (after development of the condition) (p<0.001). There was no significant difference in data obtained from patients with and without primary sclerosing cholangitis before development of the disease. CONCLUSIONS: Colonic disease subsides when primary sclerosing cholangitis develops. The higher frequency of colonic dysplasia and cancer seen in patients with primary sclerosing cholangitis can be explained by the fact that most of them have a longer duration of total colitis and fewer need total colectomy. Even though it does not seem to cause clinical problems, the colonic disease should not be ignored in these patients. PMID- 16378295 TI - Biliary stones and stenoses: diagnostic value of magnetic resonance cholangiography. AB - BACKGROUND/AIMS: To evaluate the accuracy of magnetic resonance cholangiography for detection of bile duct calculi and stenosis. METHODS: Half-Fourier single shot rapid acquisition with relaxation enhancement sequence magnetic resonance cholangiography was performed prospectively in 68 patients who were suspected of having choledocholithiasis or biliary tree stenosis. On the basis of findings at ultrasound, computed tomography, endoscopic retrograde or percutaneous cholangiography, intraoperative cholangiography or choledocoscopy and exploration, final diagnoses were normal bile ducts (n=8), choledocholithiasis (n=28), benign or malignant stenosis (n=32). RESULTS: Choledocholithiasis was diagnosed with a sensitivity of 96% and a specificity of 98%. False negative readings occurred due to stones less than two mm at in size at the distal common bile duct. A false diagnosis of choledocholithiasis (single impacted stone) by magnetic resonance cholangiography occurred in only one case for whom the final diagnosis was main bile duct adenocarcinoma, suspected on endoscopic retrograde cholangiography and confirmed at surgery. Bile duct stenosis was diagnosed with a sensitivity of 97% and a specificity of 94%. CONCLUSIONS: With magnetic resonance cholangiography, bile duct calculi and stenosis can be diagnosed with high accuracy. It is a fast, accurate and noninvasive alternative to endoscopic retrograde cholangiography in the evaluation of biliary tract disease. PMID- 16378296 TI - The effect of Helicobacter pylori eradication on symptoms and gastric emptying in patients with nonulcer dyspepsia. AB - BACKGROUND/AIMS: This study was undertaken to evaluate the effect of eradication therapy on the symptoms of Helicobacter pylori positive non-ulcer dyspepsia patients. METHODS: Twenty-four patients participated in the study and the symptoms of daytime epigastric pain, night or hunger pain, nausea, vomiting, regurgitation, bloating, belching, early satiety and anorexia were scored at the beginning, the 15th day after starting eradication therapy (amoxicillin 2 gr bid, clarithromycin 2 gr bid and omeprazole 40 mg daily for two weeks) and during the third and sixth months. Gastric emptying of radiolabelled solid meal was determined at baseline and during the third month. RESULTS: The Helicobacter pylori eradication rate was 79% and symptom scores significantly decreased during the follow-up period in both of the groups, irrespective of Helicobacter pylori status. The mean symptom scores of the 24 patients at baseline, day 15 and and months three and six were as follows: 1.275, 0.274, 0.496 and 0.238 respectively. Symptom scores for the 19 patients with Helicobacter pylori eradication were 1.084, 0.263, 0.347 and 0.215 respectively while in the five patients in whom Helicobacter pylori eradication therapy failed it was 2.0, 0.314, 1.06 and 0.32 respectively. Of the 16.6% Helicobacter pylori positive non-ulcer dyspepsia patients who had delayed gastric emptying of solids, there was no change after eradication therapy. Nine patients, including all of those in whom eradication therapy failed, required further medication (antacids/prokinetics) for continuing symptoms one month after completion of treatment. CONCLUSIONS: The results of this study suggest that Helicobacter pylori is a causal factor in symptoms of non ulcer dyspepsia and that eradication therapy improves symptoms and endoscopic findings but has no effect on gastric emptying. PMID- 16378297 TI - Relationship between gastritis severity, Helicobacter pylori intensity and mast cell density in the antrum and corpus. AB - BACKGROUND/AIMS: The aim of this study was to investigate the relationship between mast cell density, Helicobacter pylori intensity and histopathological severity of gastritis in the corpus and antrum mucosa. METHODS: The study included 59 Helicobacter pylori-positive and 20 Helicobacter pylori-negative patients. All cases underwent endoscopy, and biopsies were obtained for the evaluation of Helicobacter pylori and histopathological examination. All biopsies were evaluated according to the Sydney system and mast cell density in both the corpus and antrum mucosa was analyzed by modified Giemsa stain. Spearman's correlation test was used to determine the relationship between mast cell density and other histopathological parameters. The comparision of mast cell density between H. pylori positive and negative groups was analysed by Mann Whitney U test. RESULTS: Both in the antrum and the corpus, mast cell density was significantly higher in the Helicobacter pylori-positive group than in the Helicobacter pylori-negative group (p<0.001). The higher mast cell distribution was correlated with increased inflammation, activity and Helicobacter pylori in the antrum and corpus (p<0.001). No relationship was found between mast cell distribution and intestinal metaplasia or atrophy. CONCLUSIONS: In the light of the results of our study, mast cells may play a role in the development of Helicobacter pylori gastritis. PMID- 16378298 TI - Does eradication of Helicobacter pylori infection reduce hypergastrinema during long term therapy with proton pump inhibitors? AB - BACKGROUND/AIMS: Proton pump inhibitors and Helicobacter pylori infection are the major cause of hypergastrinemia. In this study the effect of eradication therapy on blood gastrin levels in patients using long term proton pump inhibitors was evaluated. METHODS: Twenty-seven Helicobacter pylori (positive) patients were included in the study, of whom 20 were given eradication treatment for seven days consisting of ranitidine bismuth citrate, clarithromycin and amoxicillin (eradication therapy group) and seven were given symptomatic therapy (symptomatic therapy group). Four patients who remanied Helicobacter pylori (positive) after eradication therapy were later added to the symptomatic therapy group. Lansoprazole 30 mg/day was then given to both the eradication therapy (n: 16) and the symptomatic therapy groups (n: 11) for the following three months. Fasting and non-fasting blood gastrin levels were measured initially then at one and four months after treatment. RESULTS: Fasting gastrin levels were 49 % higher in the fourth month than in the first month (p<0.01) and 51 % higher than the initial level (p<0.01) in the symptomatic therapy group. There were no statistical differences between the initial, first month and fourth month non-fasting blood gastrin levels in the symptomatic therapy group (p>0.05). Fasting gastrin levels were 47 % higher in the fourth month than the first month (p<0.001) and 18 % lower than the initial level (p<0.05) in the eradication therapy group. Non fasting gastrin levels were 4% higher in the fourth month than in the first month (p>0.05) and 34 % lower than the initial level (p< 0.05) in the eradication therapy group. CONCLUSIONS: It is suggested that patients receiving long term proton pump inhibitor treatment should be evaluated for Helicobacter pylori positivity and treatment of this infection would be an appropriate approach to avoid hypergastrinemia. PMID- 16378299 TI - Acute oligo-anuric renal failure during the course of non-fulminant hepatitis A in a patient with anorexia nervosa. AB - Acute renal failure is a very rare complication seen during the course of non- fulminant hepatitis A. Several mechanisms have been postulated in the pathogenesis of renal failure. Firstly, there is insufficiency of renal blood flow due to developing endotoxemia or cryoglobulinemia, secondly mesangial proliferative glomerulonephritis or interstitial nephritis occurs due to immune complexes and finally there is acute tubular necrosis caused by the direct cytopathic effect of the virus or due to immune complexes. The following case report describes a 17 year old male patient admitted with complaints of appetite loss and severe weight loss due to anorexia nervosa. During the second week of admission, he developed hepatitis A infection which was complicated by acute renal failure requiring hemodialysis therapy. Hepatorenal parameters returned to normal values by the fifth week of admission in this case of biopsy proven acute tubular necrosis. In this case, the possible negative effects of malnutrition on the liver and kidneys were not observed. The present authors emphasize that during the course of non- fulminant hepatitis A, renal functions should be closely monitored and renal biopsy should be performed if acute renal failure occurs. PMID- 16378300 TI - A brucellosis case with ascites, hearing loss and pancytopenia. AB - To date there has been no association of ascites, hearing loss and pancytopenia in brucellosis in the literature. A 25-year-old female patient was hospitalized with fever, hearing loss, ascites and pancytopenia. Cultures from bone marrow and ascites yielded growth of Brucella melitensis and Brucella standard tube agglutination was found to be positive at a titer of 1/1280. The patient completely recovered by the sixth week following combined antibacterial treatment of ciprofloxacin, rifampin and doxychline. PMID- 16378301 TI - Gastric heterotopia in the gallbladder: a case report. AB - A case of incidentally identified ectopic gastric mucosa in the gallbladder of a 26-year-old man with a three year history of recurrent abdominal pain is presented. The aberrant gastric tissue consisted of gastric pyloric glands and fundic glands. Focal intestinal metaplasia characterized by goblet cells containing sialomucin was seen in the surrounding gallbladder mucosa but no metaplastic change was seen in the heterotopic tissue. PMID- 16378302 TI - Primary hypertrophic pyloric stenosis in the adult: a case report. AB - A 66-year-old male was admitted with a two-month history of vomiting and weight loss. Endoscopy showed a pyloric obstruction and the patient underwent subtotal gastrectomy with gastro- jejunostomy. The histopathological study of the specimen revealed primary hypertrophic pyloric stenosis without any evidence of duodenal peptic disease. In the adult, this is a rare cause of gastric outlet obstruction of unknown etiology. It is usually recognized by histopathological examination of the specimen after a gastric resection performed to treat gastric outlet obstruction syndrome. However, some endoscopic and radiological signs, such as the cervix sign, or elongation of the pyloric channel, may give clues about the presence of the disease preoperatively. In symptomatic cases, surgery is the preferred treatment modality. PMID- 16378303 TI - Terbinafine induced prolonged cholestasis: case report and review of the literature. PMID- 16378304 TI - Diagnosis of complications in auxiliary heterotopic partial-liver transplant recipients: spiral CT findings. AB - BACKGROUND/AIMS: Imaging is important after liver transplantation to determine possible complications. The purpose of this study was to evaluate the spiral CT findings in auxiliary heterotopic partial liver transplant recipients and to determine the incidence of these complications with spiral CT imaging. METHODS: Twenty-one patients had auxiliary heterotopic partial liver transplantation at our institution during a two-year period. There were 16 males and five females with an age range of 4-58 years. All graft livers were transplanted to the right subhepatic space. Spiral CT was performed at least once after transplantation in twenty of the patients in order to evaluate vascular structures and determine possible complications. The 5mm and/or 8 mm thick sections were obtained through the abdomen before and after IV-contrast at the portal phase. Spiral CT findings were compared with ultrasonography (n=15), digital subtraction angiography (n=8), percutaneous transhepatic cholangiography (n=3) and laparotomy findings (n=2). RESULTS: Complications were detected in fourteen of the twenty-one patients. Spiral CT detected a total of 19/23 (83%) vascular complications in this study. All biliary complications (100%), 13/16 (81%) parenchymal changes and 14/14 (100%) different forms of fluid collections were also detected by spiral CT. In three patients with focal lesions, the infarcts were detected only by CT. It was not able to detect hepatic artery stenosis and one of the pseudoaneurysms of the hepatic artery. CONCLUSIONS: It is suggested that SCT in particular should be used in the evaluation of biliary complications, fluid collections and parenchymal changes of graft liver. It could be used in combination with other non-invasive imaging methods for evaluation of vascular structures. PMID- 16378305 TI - The effect of ursodeoxycholic acid treatment on epidermal growth factor in patients with bile reflux gastritis. AB - BACKGROUND/AIMS: Study was performed to evaluate the effect of ursodeoxychlic acid treatment on epidermal growth factor, which is secreted in response to mucosal injury and is also a factor in the protection and healing of gastric mucosal injury in patients with bile reflux gastritis following cholecystectomy. METHODS: Thirty-one dyspeptic patients who had previously undergone cholecystectomy were included in the study. Upper gastrointestinal endoscopy was performed before and after a six week ursodeoxychlic acid treatment period and a biopsy was taken. Endoscopic biopsy materials were stained with epidermal growth factor (Zymed, supersensitive) immunohistochemical monoclonal kit. RESULTS: The results of endoscopic examination prior to treatment were as follows: 24 cases (77%) had reflux gastritis, five cases (16%) antral gastritis, two cases (6.5%) diffuse gastritis and all cases had enterogastric reflux. In all but one case, epidermal growth factor was found to be positive at varning degrees. After ursodeoxychlic acid treatment, complete healing was observed at endoscopy in nine cases (29%) and partial healing at varning degrees was observed in all others. The degree of positivity of epidermal growth factor reduced significantly (p<0.001). CONCLUSIONS: A decrease in the degree of epidermal growth factor positivity was observed following ursodeoxychlic acid treatment. This can be explained by the decrease in epidermal growth factor release due to healing of mucosal injury following treatment. Further investigations are needed to clarify whether ursodeoxychlic acid has a direct effect on epidermal growth factor. PMID- 16378306 TI - Risk factors associated with changes in oxygenation and pulse rate during colonoscopy. AB - BACKGROUND/AIMS: Although hypoxemia is a relatively common complication of colonoscoy, the possible predictive factors of oxygen desaturation and tachycardia in patients undergoing this procedure are not well known. In this study, the possible predictive factors of severe oxygen desaturation (SaO2<90%) and tachycardia in patients with undergoing colonoscopy were investigated. METHODS: A total of 79 consecutive patients were evaluated in the study (46 men and 33 women). Significant oxygen desaturation was considered to be a reduction of arterial oxygen saturation (SaO2) to less than 90%. Tachycardia was defined as a heart rate above 100/min. Patients with inadequate colonoscopy were excluded from the study. The incidence of arterial hemoglobin oxygen desaturation and changes of heart rate during colonoscopy were evaluated and clinical factors in relation to these findings were assessed. RESULTS: The SaO2 during colonoscopy fell below 90% in 19 of the 79 patients (24.1%). The risk factors for desaturation were advanced (>60yr) age (OR: 6.03; 95% CI, 1.35-26.99), receiving sedation (OR: 11.42; 95% CI, 2.05-63.49), chronic lung disease (OR: 4.54; 95% CI and 1.40-11.68), and obesity (OR: 8.95; 95% CI, 1.17-68.55). The presence of hypertension and anemia, a history of cigarette smoking and duration of the colonoscopy had no significant effect on arterial oxygen desaturation (p>0.05). The pulse rate was raised above 100/min during colonoscopy in 26 of 79 patients (32.9%). lncrease in heart rate was found to be related to arterial oxygen desaturation (OR: 13.72; 95% CI, 2.67-70.32), anemia (OR:6.17; 95% CI, 1.15 32.91) and advanced (>60 yr) age (OR: 6.08; 95% CI, 1.62-22.81). Gender, sedation, obesity, hypertension, chronic lung disease and smoking did not affect the heart rate (p>0.05). Two patients had transitional bradyarrythmia, which had no relationship with the parameters studied. There was no incidence of significant hypoxia or change in heart rate which might have caused termination of the procedure. CONCLUSIONS: Benign and transient arterial oxygen desaturation and tachycardia may occur during colonoscopy procedure. Sedation, obesity, advanced age and chronic lung disease might contribute to these adverse events. PMID- 16378307 TI - Bullous pemphigoid and cholestatic hepatitis associated with Castleman's disease. AB - The case of a 21-yr-old woman admitted with a two-week history of icterus, fever, multiple peripheral lymphadenopathy and pruritic eruption is presented. A full evaluation including computed tomography, endoscopic retrograde cholangiography, liver, skin and lymph node biopsies and biochemical tests confirmed the diagnosis of multicentric Castleman's disease (angiofollicular lymph node hyperplasia). All symptoms improved within four weeks of commencing prednisone therapy. Castleman's disease should be considered in the differental daignosis of cholestatic hepatitis and bullous pemphigoid. PMID- 16378308 TI - Elevated carbohydrate antigen 19-9 levels in a patient with choledocholithiasis. AB - Carbohydrate antigen 19-9 (CA19-9) has been used as a tumor marker in the diagnosis and differentiation of pancreatic cancers. However, significantly high levels of CA 19-9 in the absence of pancreatic malignancy have also been reported. We present a 50-year-old woman with a common bile duct stone and cholangitis, whose CA 19-9 level of 1.500 U/ml returned to normal after definitive treatment of choledocholithiasis. PMID- 16378309 TI - Unusual presentation of hepatic vascular tumors as fulminant hepatic failure. AB - Epithelioid hemangioendothelioma and angiosarcoma of the liver are rare neoplasms of vascular origin. They can present with nonspecific symptoms such as malaise and weight loss, as well as with liver-related symptoms such as abdominal pain, tender hepatomegaly and jaundice. Portal hypertension and rarely liver failure can occur. We hereby report two cases of fulminant hepatic failure that were eventually diagnosed with epithelioid hemangioendothelioma and angiosarcoma of the liver. PMID- 16378310 TI - Hepatic epithelioid hemangioendothelioma treated with orthotopic liver transplantation: a case report. AB - Hepatic epithelioid hemangiendothelioma is a rare vascular tumor. The clinical course is unpredictable and different treatment modalities are offered depending on the patients condition. Orthotopic liver tranplantation is the choice of treatment in diffuse cases without metastases. A 32 year old woman was admitted to hospital with multiple mass lesions diagnosed by ultrasonography of the liver. Physical examination was normal except for a painless hepatomegaly, and her biochemical tests were within the normal range. Computed tomographic scanning showed the presence of multiple lesions in both lobes, some of which were accompanied by a small degree of calcification. Although these findings were suggestive of hepatic epithelioid hemangioendothelioma, ultrasonographic guided fine needle aspiration biopsy failed to diagnose the exact nature of the lesions. The diagnosis of hepatic epithelioid hemangioendothelioma was confirmed by diagnostic laparotomy and immunohistochemical examination of the specimen by FVIII-RAg, CD34 and CD 31 markers. The patient was treated by orthotopic liver transplantation and had no evidence of tumor 18 months after transplantation. The problems in differential diagnosis and treatment options are discussed in this report of the first case of this rare tumor, treated by orthotopic liver transplantation in Turkey. PMID- 16378311 TI - Alverine induced toxic hepatitis: a case report. AB - Drug-induced hepatotoxicity is an important cause of hepatocellular injury. Hepatic necrosis may range from asymptomatic elevations in transaminases to fulminant hepatic failure and death. Alverine is an antispasmodic drug which is especially used in patients with irritable bowel syndrome. Only a few cases of alverine associated hepatotoxicity have been reported previously. We present the case of a patient with alverine induced hepatotoxicity and cholestasis, which has only seldomly been reported in the literature. PMID- 16378312 TI - Two giant cavernous hemangioma caused cavernous transformation of the portal vein in a pregnant woman. AB - We report a case with two giant hemangiomas of the liver that caused cavernous transformation of the portal vein in a 45-yr-old full term pregnant woman. The patient had no serious complaints other than a mass and slight pain in the right upper quadrant and there were no abnormalities in laboratory values. Delivery was performed without complication. In the six month follow-up period, no changes were seen in the lesions. PMID- 16378313 TI - A case report of doxycycline induced esophageal and gastric ulcer. AB - It is well-established that various drugs can induce esophageal and gastric mucosal injury. Generally neither motility disorder nor an anatomical stricture is found in such cases and the cause of mucosal injury is usually found to be inadequate fluid intake with medicine or consumption just prior to bedtime. Athough there are some case reports of doxycycline induced esophageal mucosal injury, doxycycline induced gastric ulcer has been reported only once in the literature. In this case report, a 33 year old female patient who had been prescribed doxycycline for pelvic inflammatory disease is presented. She was admitted with retrosternal and epigastric pain and odynophagia, which had begun after taking the second oral dose of the drug. Endoscopic examination showed ulcers in the mid esophagus and fundus of the stomach. Histologic findings supported a drug induced etiology in these ulcers. This is only the second case in the literature where gastric ulcer has been caused by doxycycline. PMID- 16378314 TI - Ileocolic intussusception due to ileal lymphoma (Case report). AB - Intussusception occurs rarely in adults. It presents with a variety of acute, intermittent, and chronic symptoms, thus making its preoperative diagnosis difficult. In adults, intussusceptions are mostly caused by organic lesions and the majority of these lesions are benign tumours. Computed tomography scanning and endoscopy have proved to be the most useful diagnostic methods. Confirmation of diagnosis and treatment of adult intussusception is surgical, with surgical resection of the intussusception without reduction being the preferred treatment in adults, as almost half of both colonic and enteric intussusceptions are associated with malignancy. We report the case of a patient with ileocolic intussusception due to an ileal lymphoma. PMID- 16378315 TI - The new 'in-plane' view of the inter-ventricular septum. PMID- 16378316 TI - Transvaginal and three-dimensional ultrasound diagnosis of twin tubal pregnancy. PMID- 16378317 TI - The implementation of an oncologist referred, exercise self-management program for older breast cancer survivors. AB - BACKGROUND: With increased breast cancer survivor rates among older women, the negative outcomes of breast cancer treatment may linger for years. METHOD: We designed and implemented an oncologist referred, exercise self-management program to increase physical activity and health-related quality of life using a pretest posttest, single group design. We recruited 34 breast cancer survivors seen for a follow-up oncology visit at two university cancer treatment centers. Women with a mean age of 59.6 years (S.D.=66) comprised the sample. Average time since diagnosed was 3.1 years; 45% had stage I breast cancer and 55% had stage II; 62% received chemotherapy and 59% received a mastectomy. Following a baseline assessment on exercise support, self-efficacy, barriers and benefits; quality of life; and a functional performance test, subjects participated in self-management classes and received telephone support. Participants (n=30) repeated the assessment at 6-months. We compared scores between time periods using t-tests. RESULTS: Older women increased frequency of weekly, moderate physical activities (por=45 years residing in American Samoa, and women 18-44 years residing in Samoa, and a significantly lower BMI in men 18-44 years residing in Samoa. These results suggest that farm work plays an important role in regulating body size and fatness of adult Samoans and may be ideal for interventions in the Samoan archipelago. PMID- 16378332 TI - Should all pregnant diabetic women undergo a fetal echocardiography? A cost effectiveness analysis comparing four screening strategies. AB - OBJECTIVE: To determine if a policy of universal fetal echocardiography for all pregnant diabetic women is cost-effective as a screening tool for congenital heart defects. STUDY DESIGN: Using a decision-analysis model, we compared the cost-effectiveness of four screening strategies: (1) none--no ultrasound is performed; (2) selective fetal echocardiography after abnormal detailed anatomic survey; (3) fetal echocardiography for only high hemoglobin A1C, and (4) universal fetal echocardiography for all diabetics. The sensitivity and specificity for each strategy were derived by literature search. The analysis was from a societal perspective using a willingness-to-pay threshold (50,000 dollars) and a theoretic cohort of 40,000 pregnant diabetics. Costs included costs of tests and the costs of complications and of raising a child with a cardiac defect. Outcomes were reported as cost per quality-adjusted life years (QALY) gained for each congenital heart defect prevented by each strategy and the number of congenital heart defects detected. One-way, multiway and probabilistic sensitivity analyses were performed. RESULTS: Compared with the other strategies, selective fetal echocardiography after abnormal detailed anatomic survey costs less per QALY gained for cardiac defect screening. Although universal fetal echocardiography was associated with a higher detection rate for cardiac defects, it was more costly. The sensitivity analyses revealed a robust model over a wide range of values. CONCLUSION: Under the baseline assumptions, selective fetal echocardiography after an abnormal detailed anatomic survey is more cost effective compared with universal fetal echocardiography as a screening strategy for cardiac defects in pregnant diabetics. PMID- 16378334 TI - Geographical differences in physiques of male youth of age 18-20 years in China. AB - Three national surveys on the physical status of 18-, 19-, and 20-year-old male candidates for military service were carried out in six geographic regions of China in 1955, 1974, and 2001. Data from these surveys for 72,000 individuals were compared by region, by time, and by age, and estimates of incremental changes by decade were made. Overall, at all time points, males in the north and northeast areas were larger and heavier than males in the southwest. Similarly, the proportion of males in the north and northeast who were overweight was greater than in the other areas. The proportion classified as "thin" was highest in the south, southwest, and northwest, reaching 42.6% in the northwest. When urban and rural areas were compared, the mean value of every measure in every region was higher for urban youth, with all but two comparisons reaching significance (P<0.05). Differences of chest circumference among age groups were significant, as were differences in height, weight, and body mass index for 18 year-olds compared to 20-year-olds. Overall, height increments per decade were greatest for males in the southwest (average of age groups 1.79 cm, 1974 to 2001) and least in the northeast (1.08), indicating some tendency toward convergence over time. Weight increments per decade over the same time were greatest in north China (1.37 kg) and least in the northwest region (0.58 kg). PMID- 16378335 TI - High ponderal index at birth predicts high estradiol levels in adult women. AB - Inter-individual variation in levels of sex hormones results from differences in genetic, developmental, and environmental factors. We tested a hypothesis that programming of the fetal neuroendocrine axis may predispose some women to produce higher levels of steroid hormones during their menstrual cycles as adults. One hundred forty-five regularly menstruating 24- to 36- year-old women collected daily saliva samples for one menstrual cycle. Data on women's birth weights and birth lengths were obtained from medical records. A positive relationship was observed between ponderal index at birth (an indicator of nutritional status, calculated as birth weight/(birth length)(3)) and levels of estradiol (E2) in menstrual cycles, after controlling for potential confounding factors. Mean E2 was 16.4 pmol/l in the low ponderal index tertile, 17.3 pmol/l in the moderate ponderal index tertile, and 19.6 pmol/l in the high ponderal index tertile (the high ponderal index group had significantly higher E2 than both low and moderate ponderal index groups, P=0.0001). This study shows a positive association between ponderal index recorded for women at birth and levels of E2 measured during their menstrual cycles as adults. This suggests that conditions during fetal life influence adult production of reproductive hormones and may contribute to inter individual variation in reproductive function. In addition, because large size at birth is one of the factors linked with an increased risk of breast cancer, our findings provide a physiological link for the observed positive relationship between indicators of energetic conditions during fetal growth and breast cancer in women. PMID- 16378336 TI - Islands inside an island: reproductive isolates on Jerba island. AB - Jerba Island, located in South Eastern Tunisia, is inhabited by four ethnic groups: Berbers, Arabs, sub-Saharans, and Jews. All live in distinct areas, although the Arabs are also distributed all over the island. The first Arab settlement was founded in the 7th century A.D., so co-existence with Berbers has lasted for more than a millennium. Religious and cultural differences have represented an obstacle to the intermixing of these groups, and among both Arabs and Berbers marriages usually occur between members from the same extended family. Using new mtDNA data and previously described Y-chromosome STR-defined haplotypes, we tested whether this reported inbreeding would be reflected in the differentiation between Berber and Arab communities. Concerning mtDNA, the Berber group presented a greater Eurasian contribution (87%), and, surprisingly, no U6 haplotypes were found; in contrast, the Arabs showed a larger contribution of sub Saharan lineages (24%) and the U6 haplogroup amounted to 10%. Another source of evidence for the reproductive isolation of the two groups was revealed through the analysis of haplotype matching (both mtDNA and Y-chromosome), showing that matching probabilities between them is of the same order of magnitude of that observed when contrasting samples from different European countries. PMID- 16378337 TI - Developmental coordination disorder and aerobic fitness: is it all in their heads or is measurement still the problem? AB - Developmental coordination disorder (DCD) is characterized by motor inproficiency, resulting in significant impairments in social and/or academic functioning. About 5-9% of all school-age children are affected. Previous research has shown that children with DCD have lower aerobic fitness levels than children without the disorder, although the reasons for this have not been tested in the literature. A potential explanation may lie in perceived adequacy regarding performance in physical activity. Although negative perceptions of adequacy in children with DCD likely reflect an accurate appraisal of actual physical abilities, aerobic fitness tests typically require minimal coordination skills. Children who perceive themselves to be less adequate are unlikely to persist at a task and may give up sooner on these tests of endurance. Using a large community based sample of children ages 9 through 14 (n=586), we examine whether differences in aerobic fitness (assessed by performance on a 20-m shuttle run test) between children who meet the criteria for DCD (n=44) and those who do not (n=542) is due to differences in perceived adequacy toward physical activity. Our results show that one-third of the effect of DCD on VO(2) can be attributed to differences in perceived adequacy. These results suggest that at least part of the reason children perform less well on tests of aerobic endurance is because they do not believe themselves to be as adequate as other children at physically active pursuits. The implications of this for further research are discussed. PMID- 16378338 TI - African gene flow to north Brazil as revealed by HBB*S gene haplotype analysis. AB - Haplotypes linked to the HBB*S gene were analyzed in a sample of 260 chromosomes of Brazilian sickle cell anemia patients from the population of Belem, state of Para, to evaluate if the present-day haplotype frequencies correlate as well as expected with historical information on the geographic origin of African slaves sent directly to Northern Brazil. The HBB*S gene haplotype distribution (66% Bantu, 21.8% Benin, 10.9% Senegal, and 1.3% Cameroon) is in agreement with those observed for other Brazilian populations regarding the highest proportion of the Bantu type, followed by the Benin type, but it differs significantly concerning the Senegal type as this haplotype is rare or absent in samples from other Brazilian regions already studied. In addition, our results are in accordance with historical records that establish that about 90% of the slaves sent to Northern Brazil were from Angola, Congo, and Mozambique, where the Bantu haplotype predominates, in contrast to 10% of slaves from Senegambia, Guine Bissau, and Cape Verde, where the Senegal haplotype is the most common. On the other hand, the observed frequency of the Benin haplotype in Belem was much higher than that expected by historical data. This fact corroborates the suggestion that the high prevalence of the Benin type in Belem is due to domestic slave trade and later internal migrations, mainly from the Northeast, since there are no historical records of direct slave trade from Central West Africa to North Brazil. PMID- 16378339 TI - Blood pressure, seasonal body fat, heart rate, and ecological differences in Caboclo populations of the Brazilian Amazon. AB - This study compares blood pressure (BP) and related cardiovascular risk factors among three Caboclo communities from the Brazilian Amazon. Its purpose is to investigate possible risk differentials related to variable ecological settings and Western influences. Caxiuana is characterized as a more "traditional" group, while Aracampina and Santana are viewed as more "transitional" in lifestyle. A total of 348 subjects from the three communities were evaluated in the wet or the dry season or in both. Measurements across the communities were compared by season and sex. Results suggest little seasonal variation in average BP, BP change, body fat, or body fat change among men. Conversely, there is substantial seasonal and inter-community variation among women. Additional analyses reveal (1) an inconsistent association between age and BP across the communities; (2) that BMI is not associated with BP transitional communities in either season but is associated with both systolic and diastolic pressure in the most traditional community; and (3) little to no sex effect on BP. These results suggest increased Western influence affects body composition particularly of women. However, increased BMI and fat among transitional Caboclo women does not directly translate into higher BP; rather, their BP appears to be more affected by seasonal stresses. Finally, conditions during the wet season diminish age-related variation in BP, suggesting that during the wet season these Caboclo may be less active. PMID- 16378340 TI - Y-chromosome diversity is inversely associated with language affiliation in paired Austronesian- and Papuan-speaking communities from Solomon Islands. AB - The Solomon Islands lie in the center of Island Melanesia, bordered to the north by the Bismarck Archipelago and to the south by Vanuatu. The nation's half million inhabitants speak around 70 languages from two unrelated language groups: Austronesian, a language family widespread in the Pacific and closely related to languages spoken in Island Southeast Asia, and "East Papuan", generally defined as non-Austronesian and distantly related to the extremely diverse Papuan languages of New Guinea. Despite the archipelago's presumed role as a staging post for the settlement of Remote Oceania, genetic research on Solomon Island populations is sparse. We collected paired samples from two regions that have populations speaking Austronesian and Papuan languages, respectively. Here we present Y-chromosome data from these samples, the first from Solomon Islands. We detected five Y-chromosome lineages: M-M106, O-M175, K-M9*, K-M230, and the extremely rare clade, K1-M177. Y-chromosome lineages from Solomon Islands fall within the range of other Island Melanesian populations but display markedly lower haplogroup diversity. From a broad Indo-Pacific perspective, Y-chromosome lineages show partial association with the distribution of language groups: O M175 is associated spatially with Austronesian-speaking areas, whereas M-M106 broadly correlates with the distribution of Papuan languages. However, no relationship between Y-chromosome lineages and language affiliation was observed on a small scale within Solomon Islands. This pattern may result from a sampling strategy that targeted small communities, where individual Y-chromosome lineages can be fixed or swept to extinction by genetic drift or favored paternal exogamy. PMID- 16378341 TI - Longitudinal follow-up of fitness during childhood: interaction with physical activity. AB - Health-related fitness values during childhood (from age 11 to 16 years) were analyzed in relation to changes in physical activity level. One hundred fifty eight children were monitored over a 4-year period. Twice a year, they performed six physical fitness tests: standing broad jump, 10 x 5-meter shuttle run, sit and-reach, handgrip, number of sit-ups in 30 s, and 20-m shuttle run. Height was measured four times per year to assess age at peak height velocity. Physical activity was assessed with a self-administered questionnaire at baseline and at the end of the follow-up. The physical activity level was defined by the frequency and the time spent in moderate-to-vigorous physical activities. Two groups were constituted: regularly active and sedentary. A multiple linear multilevel regression analysis was used to analyze the longitudinal relationships between changes in physical activity and physical fitness levels. Corrections were made for both time-dependent (time) and time-independent (sex) variables. Except for flexibility, boys' fitness performances increased more than that for the girls. Positive and significant (P < 0.05) regression coefficients were found with the regularly active for standing broad jump, 20-m shuttle run, number of sit-ups, 10 x 5-m shuttle run in both sexes, and for the girls' sit-and-reach performance. Increasing or decreasing physical activity level was not associated with changes in fitness performances over time, except for flexibility for the girls and the 20-m shuttle run for the boys. From childhood to adolescence, increasing physical activity is not sufficient to be more fit. The children who are stayed the most active were the fittest, particularly the girls. Am. J. Hum. Biol. 18:51-58, 2006. (c) 2005 Wiley-Liss, Inc. PMID- 16378342 TI - Age-related changes in testosterone and SHBG among Turkana males. AB - To determine age-related changes in biologically available testosterone (T) among men in a subsistence society and their relationship to energetic status, T, sex hormone binding globulin (SHBG) and anthropometric measures were compared among nomadic and settled Turkana pastoralists of northern Kenya. Hormonal measures were available for 104 nomadic men and 72 settled men, estimated ages 20-90 years. Comparison of the two subpopulations revealed significantly higher blood T (32.7+/- 15.1 vs. 23.4+/-15.2 nM) and SHBG (53.8+/- 19.5 vs. 39.7+/- 20.nM) but not free testosterone index (FTI) (65.6+/- 39.3 vs.66.3+/- 45.9) among the nomads. Total blood T did not exhibit a significant linear decline with age in either subgroup, while SHBG values showed a significant linear increase among the nomads. When controlled for energetic status, FTI showed a significant decrease with age among the nomads, but not the settled males. Total blood T was negatively associated with waist circumference among the nomads, but not the settled males. FTI showed a marginally significant negative association with waist circumference, suprailiac skinfold, and % body fat among the nomads but no associations with body composition among the settled group. These results add additional evidence that T is related to energetic status under conditions of negative energy balance and suggest that cross-population variation in the slope of age-related declines in free serum T and salivary T may be related to energetic status through the effects of SHBG. PMID- 16378343 TI - Biocultural approaches in human biology. AB - Biocultural approaches recognize the pervasiveness and dynamism of interactions between biological and cultural phenomena, and they explicitly strive to integrate biological, sociocultural, environmental, and other kinds of data. They have been part of human biology at least since 1958, when Frank Livingstone so elegantly explained the linkages among population growth, subsistence strategy, and the distribution of the sickle cell gene in West Africa. These approaches developed further with the advent of human adaptability studies in the 1960s as part of the Human Biological Program and have become increasingly focused on understanding the impacts of everyday life on human biological variation. Biocultural approaches generate explanations that are intuitively appealing to many because they offer a kind of holistic view. They can, however, be very challenging approaches to implement, perhaps in part because we are more experienced in measuring the biological than the cultural. Some of the challenges include (1) defining precisely what we mean by constructs like socioeconomic status, poverty, rural, and urban; (2) operationalizing key variables so that they can be measured in ways that are ethnographically valid as well as replicable; (3) defining and measuring multiple causal pathways. In this paper, I briefly review the history of biocultural approaches and then illustrate some of the challenges that these approaches present with examples from my own research on nutrition and energetics as well as that of other practitioners. PMID- 16378344 TI - A revertant of the major founder Native American haplogroup C common in populations from northern South America. AB - We examined the mtDNA RFLP diversity of 17 Native American populations from Colombia. Five of the populations studied were found to have variable frequencies of a mtDNA type lacking the characteristic changes of haplogroups A-D. Sequencing of mtDNA HVS-I and II showed that this "null" RFLP type carries all the substitutions characteristic of Native American founder lineage C. A back mutation has therefore recreated the +13,259 HincII/-13,262 AluI restriction sites that tipify RFLP haplogroup C. This revertant C lineage is further characterized by three changes in HVS-II sequence: C/T transitions at positions 115 and 152, and the deletion of an A residue at position 116. This lineage is observed at high frequency mostly in populations from Greenberg's Equatorial Tucano linguistic family. Genetic structure analyses are consistent with the reversion mutation occurring at an early stage during the tribalization process. PMID- 16378345 TI - Plasma amino acids in Greenlanders and Danes: influence of seasons, residence, ethnicity, and diet. AB - Basal metabolic rate is elevated among circumpolar populations. It has been our hypotheses that this is reflected in the levels of plasma amino acids, that amino acid concentration in plasma differs between Greenlanders and Danes, and that this difference is related to residence, ethnicity, diet, and season. The purpose of the study was to measure plasma amino acids in Greenlanders and Danes and to analyze the influence of residence, ethnicity, diet, and season. Amino acids in plasma were measured in four groups of healthy subjects both during summer and winter: Group 1, Danes living in Denmark consuming European food; Group 2, Greenlanders living in Denmark consuming European food; Group 3, Greenlanders living in Greenland consuming European food; and Group 4, Greenlanders living in Greenland consuming mainly traditional Greenlandic food. Amino acids were determined by pre-column derivatization with o-phthaldialdehyde and reverse-phase high-performance liquid chromatography with gradient elution and fluorescence detection. Most plasma amino acids were lower during summer than winter in Greenlanders living in Greenland. Comparison of the four groups showed that residence in Greenland was the most important influencing factor for the concentration of plasma amino acids, whereas ethnicity and diet had only a very modest or no effect. These findings could not be attributed to changes in thyroid function. However, the level of physical activity was significantly higher in Groups 3 and 4 than in Groups 1 and 2. Because exercise reduces the amino acid pool in plasma, it is possibly that the higher physical activity among Greenlanders living in Greenland explains the reduction in plasma amino acids during summer. It is concluded that plasma amino acids were lower during summer than winter in Greenlanders living in Greenland compared with Greenlanders in Denmark and Danes. This difference might be due to the higher level of physical activity among Greenlanders in Greenland during the summer period. PMID- 16378346 TI - Predictors of South African male testosterone levels: the THUSA study. AB - The process of urbanization occurring in many developing countries may have consequences for reproductive endocrine function. Here, we test predictions concerning variation in South African male testosterone levels among subgroups across an urbanization gradient representing differences in both geography and socioeconomic status. Subjects included 364 males aged between 20 and 82 comprising a cross-sectional study conducted between 1996 and 1998. Testosterone levels were measured from serum samples obtained between 08:00 and 11:00. In ANCOVA analysis, male testosterone levels differed significantly along this rural to-urban gradient, with members of the most urban group having higher testosterone levels than groups of farmers and inhabitants of informal housing areas adjacent to towns. Testosterone levels declined with age and were negatively related to body mass index (BMI). Testosterone levels did not differ according to HIV status. Further exploratory ANCOVA analyses revealed that physical activity levels, depression, affect, and hostility were not significantly associated with variation in testosterone levels. These data help document causes of variation in male testosterone levels in a context of urbanization and may have implications for clinical outcomes such as the development of a male hormonal contraceptive or prostate cancer. PMID- 16378347 TI - Study of GM immunoglobulin allotypic system in Berbers and Arabs from Morocco. AB - The GM immunoglobulin allotype polymorphism was investigated in four Moroccan populations: three Berber groups from Khenifra (Middle Atlas), Amizmiz (High Atlas), and Bouhria (Beni Snassen) and one Arabic-speaking sample from the Doukkala area (Abda, Chaouia, Doukkali, and Tadla districts in south-central Morocco). In order to characterize the genetic relationships between the populations, our results were compared with those obtained for other North African groups (from Morocco, Algeria, Tunisia, and Niger) and for Middle-East Africans, sub-Saharans, and Southwest Europeans. Based on GM haplotype frequencies, Factorial Correspondence Analyses, F(ST) significance testing, and hierarchical analyses of variance were performed. Our results reveal that Moroccan populations have heterogeneous GM profiles with high frequencies of GM haplotypes in Europeans (from 76% for Doukkala to 88% for Bouhria) and relatively high frequencies of GM haplotypes in sub-Saharans (from 11% for Bouhria to 23% for Amizmiz). The genetic diversity observed among Moroccans is not significantly correlated with either geographic or linguistic differentiation. In spite of their cultural and historical differentiation, we did not discover any significant genetic differences between Berbers and Arabic-speakers from Morocco. However, when large geographical areas are considered, our population samples are integrated in the North African GM variation, significantly distant from sub Saharan groups but with a close relationship with Southwest European populations. PMID- 16378348 TI - Secular trends in age at menarche among Caboclo populations from Para, Amazonia, Brazil: 1930-1980. AB - The present study is part of a large research project on growth, development, nutrition, and health of Caboclo populations from the Brazilian Amazon. The aim of this paper is to analyze the age of menarche in adolescents (N=164) and adult women (219) in the studied populations. Caboclo are admixed rural, peasant groups that live along the Amazon river and its tributaries, and there are few previous studies about them. Probit analysis of the status quo data yielded a median age at menarche of 12.29+/-1.76 years. The retrospective method was applied to recall data of the 77 post-menarcheal adolescents, yielding an average of 13.06+/-1.27 years. Number of children in the family did not show any statistical influence on age at menarche in any age group. In adult women, age at menarche decreased from 14.50 in those born in 1930 to 12.88 for those born in 1980 (F=4.371, P=0.001). The downward trend found was, on average, 0.237 year per decade in the study period. The median age at menarche in the adolescents (12.29 years) is one of the lowest values found for Central and South American populations. In the ecological context, a low age at menarche could be an adaptive advantage because it provides a greater chance for reproduction at a young age in an environment where, until recently, life expectancy was low. As has been reported for other developing countries, the change found in age at menarche in the women born from the 1930s to the 1980s is likely to be related to changes in health and nutritional factors that occurred in Brazil because this country experienced significant improvement in living standards related to education, vaccination, and health conditions, which, although not equally, reached all regions after the 1960s. PMID- 16378350 TI - Healthy eating. Get saucy! Health alternatives to high-fat toppings. PMID- 16378349 TI - Individual-, neighborhood-, and state-level socioeconomic predictors of cervical carcinoma screening among U.S. black women: a multilevel analysis. AB - BACKGROUND: Cervical carcinoma is the fifth most common cancer among African American women in the U.S. Although the Papanicolaou (Pap) smear is an efficacious screening tool in the early detection of the disease, disparities are known to persist in the utilization of this procedure across socioeconomic groups. METHODS: Data regarding cervical carcinoma screening and covariates were obtained from the 59,090 Black Women's Health Study participants across the U.S. via a mailed questionnaire in 1995. Logistic regression and multilevel techniques were used to assess the independent effects of the covariates on nonrecent cervical carcinoma screening. RESULTS: In all, 8.3% of the 40,009 women in the present analysis had not undergone a Pap smear examination within the previous 2 years (nonrecent screening). Lower educational attainment, older age, obesity, smoking, and neighborhood poverty were found to be independently related to increased risk of nonrecent screening. The adjusted odds ratio for nonrecent screening was 1.2 (95% confidence interval [95% CI], 1.1-1.4) for women residing in neighborhoods with 20% or more poverty compared with those in neighborhoods with less than 5% poverty. State of residence was also associated with nonrecent cervical carcinoma screening. CONCLUSION: These results suggest that among black women, residence in high-poverty (20%) neighborhoods is associated with an increased risk of nonrecent cervical carcinoma screening, independent of individual level risk factors. PMID- 16378351 TI - Eye disease. Understanding retinopathy. PMID- 16378352 TI - Stress and glucose levels. PMID- 16378353 TI - Underdiagnosed neuropathy. PMID- 16378354 TI - Adventures on the Internet--the quest for information. PMID- 16378355 TI - Foot surgery (save your leg and your life). PMID- 16378357 TI - Warm your insides. Tasty winter suppers that are a snap to prepare. PMID- 16378356 TI - Can't feel your lows? Understanding hypoglycemia unawareness. PMID- 16378359 TI - Research profile. Blood vessel damage. Seeking the diabetes connection. Marc. K. Halushka, MD, PhD. PMID- 16378358 TI - Women, polycystic ovary syndrome, and type 2--we are family. This "sister" you never knew you had may be wreaking havoc with your hormone and insulin levels. PMID- 16378360 TI - An endophytic fungus from Nothapodytes foetida that produces camptothecin. AB - A fungal endophytic isolate, camptothecin, has been isolated from the inner bark of the plant Nothapodytes foetidafrom the Western coast of India. The fungus, which belongs to the family Phycomycetes, produced the anticancer drug lead compound camptothecin (1) when grown in a synthetic liquid medium (Sabouraud broth) under shake flask and bench scale fermentation conditions. Compound 1 was identified by means of chromatographic and spectroscopic methods. It was also compared with an authentic example for its biological activity against a number of human cancer cell lines. Isolation of an organism producing 1 and its fermentation may, in the future, provide an easily accessible source for the production of this anticancer drug precursor molecule. PMID- 16378361 TI - An anthraquinone with potent quinone reductase-inducing activity and other constituents of the fruits of Morinda citrifolia (noni). AB - Morinda citrifolia, commonly known as noni, has a long history of utilization throughout much of tropical Polynesia and is considered to be the second most important medicinal plant in the Hawaiian Islands. Recently, the use of noni as a dietary supplement in the United States has greatly increased. Bioassay-guided fractionation of a dichloromethane-soluble partition of a MeOH extract of noni fruits has led to the isolation of an extremely potent quinone reductase inducer, 2-methoxy-1,3,6-trihydroxyanthraquinone (1). This new anthraquinone (1) was nearly 40 times more potent than a positive control, l-sulforaphane. Furthermore, compound 1 demonstrated no discernible cytotoxicity at the highest dose tested. In addition to compound 1, 11 known compounds were also isolated and identified in the present investigation. This is the first report of the isolation of anthraquinones from noni fruits. PMID- 16378362 TI - Fluorescent compounds from the cultured mycobiont of Amygdalaria panaeola. AB - New fluorescent compounds, named panaefluorolines D-H, were isolated from the cultured mycobiont of a lichen, Amygdalaria panaeola. The structures were elucidated on the basis of spectroscopic data, especially 2D-NMR. The relative configuration of panaefluoroline D was determined by means of X-ray crystallographic analysis. PMID- 16378363 TI - Antiviral Terpenoid Constituents of Ganoderma pfeifferi. AB - Four sterols and 10 triterpenes were isolated from the fruiting bodies of Ganoderma pfeifferi, including the three new triterpenes 3,7,11-trioxo-5alpha lanosta-8,24-diene-26-al (lucialdehyde D, 1), 5alpha-lanosta-8,24-diene-26 hydroxy-3,7-dione (ganoderone A, 2), and 5alpha-lanosta-8-ene-24,25-epoxy-26 hydroxy-3,7-dione (ganoderone C, 3). The structures of 1-3 were determined on the basis of spectroscopic evidence. Antibacterial, antifungal, and antiviral activity were studied for some of the isolated compounds. Ganoderone A (2), lucialdehyde B (4), and ergosta-7,22-dien-3beta-ol (7) were found to exhibit potent inhibitory activity against herpes simplex virus. PMID- 16378364 TI - Sesquiterpene chromones from Ferula fukanensis and their nitric oxide production inhibitory effects. AB - Five new sesquiterpene chromone derivatives, fukanefurochromones A-E (1-5), were isolated from a 80% aqueous methanol extract of the roots of Ferula fukanensis. The structures were elucidated on the basis of spectroscopic evidence, especially heteronuclear multiple-bond connectivity (HMBC) and high-resolution MS. The sesquiterpene chromone derivatives inhibited nitric oxide (NO) production and inducible NO synthase (iNOS) gene expression by a murine macrophage-like cell line (RAW 264.7), which was activated by lipopolysaccharide (LPS) and recombinant mouse interferon-gamma (IFN-gamma). PMID- 16378365 TI - Hygrocins a and B, naphthoquinone macrolides from Streptomyces hygroscopicus. AB - Two new naphthoquinone macrolides, hygrocins A (1) and B (2), were isolated from the fermentation broth of Streptomyces hygroscopicus. Hygrocin A is not stable due to the presence of an active methylene group (C-22), which undergoes intramolecular aldol condensation with the quinone ring to yield a gamma-lactam derivative, 6. Its structural elucidation was achieved by chemical conversion to 3, an unusual diazomethane derivative, and confirmed by its alkaline hydrolysis product 4, hydrogenation derivative 5, and "degradation" product 6. The structure of hygrocin B was determined by combined chemical and spectroscopic methods. PMID- 16378366 TI - A revised structure for alkaloid 235C isolated from skin extracts of mantellid (Mantella) frogs of Madagascar. AB - Madagascan frogs of the mantellid genus Mantella have been a rich source of alkaloids derived from dietary arthropods. Two species of frogs, inhabiting swamp forest, contain a unique set of alkaloids, previously proposed, based only on GC MS and GC-FTIR data, to represent dehydro analogues of the homopumiliotoxins. The major alkaloid of this set, alkaloid 235C (2), now has been isolated in sufficient quantities (ca. 0.3 mg) to allow determination of the structure by NMR analysis. The structure of alkaloid 235C proved to be a 7,8-dehydro-8 desmethylpumiliotoxin. A comparison is presented between the mass, infrared, and (1)H NMR spectra of 235C (2) and a synthetic dehydrohomopumiliotoxin (1), initially proposed incorrectly as the structure for 235C. PMID- 16378367 TI - Sesquiterpenoids and norsesquiterpenoids from the Formosan soft coral Lemnalia laevis. AB - Eight new nornardosinane sequiterpenoids, laevinols A-H (1-8), a new neolemnane sesquiterpenoid, laevinone A (9), and the previously known 6beta-acetyl 4beta,5beta-dimethyl-1(10)alpha-epoxy-2beta-hydroxy-7-oxodecalin (10) and 11,12 dihydroxy-6,10-eremophiladiene (11) were isolated from the methylene chloride solubles of the Formosan soft coral Lemnalia laevis. Their structures were elucidated by extensive spectroscopic analysis, and their cytotoxicity against selected cancer cells was measured in vitro. PMID- 16378368 TI - Cucurbitosides F-M, acylated phenolic glycosides from the seeds of Cucurbita pepo. AB - Eight new phenolic glycosides, cucurbitosides F-M (1-8), were isolated from the seeds of Cucurbita pepo. Their structures were elucidated as 4-(2 hydroxyethyl)phenyl 5-O-(2-S-2-methylbutyryl)-beta-d-apiofuranosyl(l-->2)-beta-d glucopyranoside (1), 4-(2-hydroxyethyl)phenyl 5-O-(3-methylbutyryl)-beta-d apiofuranosyl(l-->2)-beta-d-glucopyranoside (2), 4-(2-hydroxyethyl)phenyl 5-O nicotinyl-beta-d-apiofuranosyl(l-->2)-beta-d-glucopyranoside (3), 4-(2 hydroxyethyl)phenyl 5-O-(4-aminobenzoyl)-beta-d-apiofuranosyl(l-->2)-beta-d glucopyranoside (4), 4-(2-hydroxyethyl)-2-methoxyphenyl 5-O-(2-S-2-methylbutyryl) beta-d-apiofuranosyl(l-->2)-beta-d-glucopyranoside (5), 4-(hydroxymethyl)phenyl 5 O-(2-S-2-methylbutyryl)-beta-d-apiofuranosyl(l-->2)-beta-d-glucopyranoside (6), 4 (hydroxymethyl)phenyl 5-O-nicotinyl-beta-d-apiofuranosyl(l-->2)-beta-d glucopyranoside (7), and 4-(hydroxymethyl)phenyl 5-O-(4-aminobenzoyl)-beta-d apiofuranosyl(l-->2)-beta-d-glucopyranoside (8) on the basis of various spectroscopic analyses and analyses of hydrolysis products. PMID- 16378369 TI - ent-Kaurane Diterpenoids from Isodon albopilosus. AB - Nine new diterpenoids, albopilosins B-J (1-9), together with six known analogues, albopilosin A (10), macrocalyxin C (11), rabdokunmin C (12), excisanin (13), amethystonoic acid (14), and coetsanoic acid (15), were isolated from the aerial parts of Isodon albopilosus. The structures of 1-9 were established using spectroscopic methods including extensive 1D and 2D NMR analysis. The diterpenoids isolated were evaluated for their inhibitory activities against HepG2 (hepatoma) cells. Compounds 7 and 13 were the most active, with both having IC(50) values of <15 microM. PMID- 16378371 TI - Botcinins A, B, C, and D, metabolites produced by Botrytis cinerea, and their antifungal activity against Magnaporthe grisea, a pathogen of rice blast disease. AB - Four new metabolites, botcinins A-D, were isolated from the culture filtrate of a strain of Botrytis cinerea. Their structures were determined by spectroscopic methods, mainly NMR techniques, molecular modeling, and the modified Mosher's method. They exhibited antifungal activities against Magnaporthegrisea, a pathogen of rice blast disease. Botcinins B and C have a MIC of 12.5 microM, and botcinins A and D are not active below 100 microM. PMID- 16378370 TI - Phytochemical studies on Stemona plants: isolation of stemofoline alkaloids. AB - Six new stemofoline alkaloids, (2'R)-hydroxystemofoline (5), (3'R)-stemofolenol (6), (3'S)-stemofolenol (7), 1',2'-didehydrostemofoline-N-oxide (8), the first C(19) stemofoline alkaloid, methylstemofoline (9), and the first glycosidated Stemona alkaloid, stemofolinoside (10), and three known alkaloids, (2'S) hydroxystemofoline (2), (11Z)-1',2'-didehydrostemofoline (3), and (11E)-1',2' didehydrostemofoline (4), have been isolated from a root extract of an unidentified Stemona species. The structure and relative configuration of these new alkaloids have been determined by spectral data interpretation and from semisynthetic studies. PMID- 16378372 TI - Flavonoids and benzene derivatives from the flowers and fruit of Tetrapanax papyriferus. AB - Two new flavonoids, kaempferol 7-O-(2-E-p-coumaroyl-alpha-l-rhamnoside) (1) and kaempferol 7-O-(2,3-di-E-p-coumaroyl-alpha-l-rhamnoside) (2), together with 10 known compounds were isolated from the flowers and fruit of Tetrapanax papyriferus. Compounds 1 and 2 showed cytotoxicity by brine shrimp lethality bioassay with LC(50) values of 0.57 and 0.40 mM, respectively. PMID- 16378373 TI - Perspicamides A and B, quinolinecarboxylic acid derivatives from the Australian ascidian Botrylloides perspicuum. AB - Two novel quinoline-2-carboxylic acid derivatives, perspicamides A (2) and B (3), were isolated from the Australian ascidian Botrylloides perspicuum. Extraction of the freeze-dried ascidian with methanol and purification of the extract by C18 MPLC followed by repeated C18 HPLC yielded the new compounds 2 and 3 as well as the known compounds botryllamides A-C (4-6). The structures of 2 and 3 were determined from analysis of 2D NMR spectra. PMID- 16378374 TI - Bis(bibenzyls) from liverworts inhibit lipopolysaccharide-induced inducible NOS in RAW 264.7 cells: a study of structure-activity relationships and molecular mechanism. AB - The inhibition of lipopolysaccharide-induced NOS by 19 bis(bibenzyls) isolated from liverworts in RAW 264.7 macrophages was evaluated. The presence of phenolic hydroxyls and saturation at 7,8 and/or 7'/8' are required for inhibition of NO production. Among the compounds tested, marchantin A was the most potent, and its inhibitory activity was consistent with the inhibition of LPS-induced iNOS mRNA. PMID- 16378375 TI - Antimycobacterial scalarane-based sesterterpenes from the Red Sea sponge Hyrtios erecta. AB - A new scalarane-type pentacyclic sesterterpene, sesterstatin 7 (1), was isolated from the Red Sea sponge Hyrtios erecta, together with 16-epi-scalarolbutenolide (2), 25-dehydroxy-12-epi-deacetylscalarin (3), 3-acetylsesterstatin 1 (4), and 21 acetoxydeoxyscalarin. The structure of 1 was elucidated by spectroscopic data interpretation. Sesterstatin 7 (1) showed 63% inhibition of Mycobacterium tuberculosis (H(37)Rv) at a concentration of 6.25 microg/mL. Compound 2 displayed moderate inhibitory activity, while 3 and 4 were weakly active against the same biological target. PMID- 16378376 TI - Acryloylated glucose 3-nitropropanoates from Indigofera kirilowii. AB - Two unique glucose esters of 3-nitropropanoic acid with acryloyl moieties, kirilowin A and kirilowin B, were isolated from the roots of Indigofera kirilowii. Their structures were elucidated by detailed spectroscopic analysis. The two acrylates constitute new additions to the toxic glucose conjugates of 3 nitropropanoic acid in angiosperms. PMID- 16378377 TI - Dammarane triterpenes from the hypocotyls and fruits of Ceriops tagal. AB - Three new dammarane triterpenes, cereotagaloperoxide, cereotagalol A, and cereotagalol B, together with four known dammarane triterpenes, an oleanane triterpene, and 13 known lupane triterpenes were isolated from the hypocotyls and fruits of Ceriops tagal. The structures of 1-3 were characterized on the basis of their spectroscopic data. PMID- 16378378 TI - Antifungal flavonoids from Hildegardia barteri. AB - A new isoflavan, (3R)-6,2'-dihydroxy-7-methoxy-4',5'-methylenedioxyisoflavan, hildegardiol (1), and two known flavonoids, 2-hydroxymaackiain (2) and farrerol (3), were isolated from the antifungal root extract of Hildegardia barteri. The pterocarpan 2 was largely responsible for the observed antifungal activity. PMID- 16378379 TI - Nostocarboline: isolation and synthesis of a new cholinesterase inhibitor from Nostoc 78-12A. AB - A new quaternary beta-carboline alkaloid, nostocarboline, was isolated from the freshwater cyanobacterium Nostoc 78-12A, and its constitution was assigned by 2D NMR methods. The structure was proven by its total synthesis starting from norharmane via chlorination at C-6 and methylation at N-2. Nostocarboline was found to be a potent butyrylcholinesterase (BChE) inhibitor, with an IC(50) of 13.2 microM. The related 2-methylnorharmane, which is present in the human brain and might be relevant to Parkinson's disease (PD), was also determined to be a BChE inhibitor (11.2 microM). These inhibitory concentrations are comparable to galanthamine, an approved drug for the treatment of Alzheimer's disease (AD). Nostocarboline can thus be considered as a lead for the development of novel neurochemicals. PMID- 16378380 TI - Discorhabdin W, the first dimeric discorhabdin. AB - Discorhabdin W (1), isolated from a New Zealand Latrunculia sp. sponge, is a symmetrical dimer with potent in vitro activity against the P388 murine leukemia cell line. The structure and stereochemistry were assigned by using 1D- and 2D NMR experiments and mass spectrometry. Discorhabdins B (2), D (3), G/I (4), and L (5) were also isolated from the same sponge. PMID- 16378381 TI - Dichlorinated pulvinic acid derivative from a Malaysian Scleroderma sp. AB - A new dichlorinated pulvinic acid derivative, methyl-3',5'-dichloro-4,4'-di-O methylatromentate, was isolated from the fruiting body of a Scleroderma sp. The structure was determined using spectroscopic methods, and an X-ray analysis was carried out for confirmation of the structure. Compound was found to display moderate antimicrobial activity against Bacillus subtilis. PMID- 16378382 TI - Xanthone O-glycosides and benzophenone O-glycosides from the roots of Polygala tricornis. AB - A new benzophenone O-glycoside, tricornoside A, and five new xanthone O glycosides, tricornosides B-F, were isolated from the roots of Polygala tricornis together with three known glycosides. The structures of new compounds were elucidated on the basis of chemical and spectroscopic evidence. PMID- 16378383 TI - Bisabolane- and santalane-type sesquiterpenoids from Santalum album of Indian origin. AB - Six new bisabolane-type (1-3) and santalane-type (4-6) sesquiterpenoids, together with (+)-alpha-nuciferol, (+)-citronellol, and geraniol, were isolated from the heartwood of Santalum album of Indian origin. Their structures, including two bisabolol diastereomers (1, 2), were established on the basis of spectroscopic data interpretation. PMID- 16378384 TI - Lycopodatines A-C, C(16)N alkaloids from Lycopodium inundatum. AB - Three new alkaloids, lycopodatines A (1), B (2), and C (3), have been isolated from the club moss Lycopodium inundatum, and the structures and absolute configuration were elucidated on the basis of 2D NMR data and chemical transformation. PMID- 16378385 TI - Metabolomics reviewed: a new "omics" platform technology for systems biology and implications for natural products research. AB - Metabolomics is the study of global metabolite profiles in a system (cell, tissue, or organism) under a given set of conditions. The analysis of the metabolome is particularly challenging due to the diverse chemical nature of metabolites. Metabolites are the result of the interaction of the system's genome with its environment and are not merely the end product of gene expression but also form part of the regulatory system in an integrated manner. Metabolomics has its roots in early metabolite profiling studies but is now a rapidly expanding area of scientific research in its own right. Metabolomics (or metabonomics) has been labeled one of the new "omics", joining genomics, transcriptomics, and proteomics as a science employed toward the understanding of global systems biology. Metabolomics is fast becoming one of the platform sciences of the "omics", with the majority of the papers in this field having been published only in the last two years. In this review metabolomic methodologies are discussed briefly followed by a more detailed review of the use of metabolomics in integrated applications where metabolomics information has been combined with other "omic" data sets (proteomics, transcriptomics) to enable greater understanding of a biological system. The potential of metabolomics for natural product drug discovery and functional food analysis, primarily as incorporated into broader "omic" data sets, is discussed. PMID- 16378388 TI - Modification of gold nanorods using phosphatidylcholine to reduce cytotoxicity. AB - Hexadecyltrimethylammonium bromide (CTAB), which is necessary for the preparation of gold nanorods (NRs), was extracted from a NR solution into a chloroform phase containing phosphatidylcholine (PC). After three extractions, the zeta potential of the NRs remained positive, but its magnitude decreased from +67 +/- 1 to +15 +/- 1 mV. Transmission electron microscopy and energy-dispersive X-ray analysis indicated that the NRs were passivated with PC. The PC layer on the NR surface contributed to the prevention of NR aggregation. The PC-passivated NRs showed low cytotoxicity in comparison with twice-centrifuged NRs. It was shown that a negligible amount of CTAB was dispersed in the NR solution after the extraction. The extraction using a chloroform phase containing PC was found to be a convenient way of replacing the CTAB with alternative capping agents such as PC. This is a key technique for preparing functional NRs that can have practical applications. PMID- 16378389 TI - Synthesis of hollow silica spheres with mesostructured shell using cationic anionic-neutral block copolymer ternary surfactants. AB - Hollow silica spheres with mesostructured shells (HSSMS) were prepared with a vesicle template of cetyltrimethylammonium bromide-sodium dodecyl sulfate Pluronic P123 (C(16)TMAB-SDS-EO(20)PO(70)EO(20)) at a SDS/C(16)TMAB ratio of 0.6 0.8 following a fast silicification in dilute silicate solution at pH approximately 5.0. The mesostructure of the shell is disordered, and the mesopore size is about 5.5-7.5 nm. Moreover, the direction and length of the nanochannels of the shell change with the SDS/C(16)TMAB ratios. A bi-template model, in which the C(16)TMA(+)-DS(-) form the stable bilayer vesicle structure and the P123 copolymers anchored on C(16)TMA(+)-DS(-) vesicle act as the template for the mesoporous silica, was proposed to explain the formation of the HSSMS. This bi template model can be applied extensively to prepare the HSSMS with different diameters and pore sizes by using other C(n)TMAX-SDS-EO(n)PO(m))EO(n) ternary surfactant mixtures. PMID- 16378390 TI - Reaction growth of MF2/a-C (M = Ca, Mg) core/shell nanowires at the interface of vapor and solid reactants. AB - C(6)F(6) vapor is employed to react with CaC(2) and Mg(3)N(2) to grow CaF(2)/a-C and Mg(2)F(2)/a-C core/shell nanowires (tens of micrometers in length, tens to hundreds of nanometers in wire diameter, and tens of nanometers in core diameter), respectively, in high yields. The growth mechanism is proposed to proceed via a reaction at the interface of the vapor and solid reactants. PMID- 16378391 TI - Characterization of surface-confined alpha-synuclein by surface plasmon resonance measurements. AB - Urea-driven denaturation and renaturation of surface-bound alpha-synuclein are monitored by surface plasmon resonance (SPR) spectroscopy. The differential SPR angle shift (Delta Theta(SPR))(Net) enables us to estimate the Gibbs free energy change (DeltaG(o)) for the denaturation of the supported alpha-synuclein. DeltaG(o) for the denaturation of the supported alpha-synuclein, which is indirectly related to its biological activity can be increased significantly by the mixed self-assembled monolayers of 11-mercaptoundecanoic acid and 1,6 hexanedithiol. These SPR measurements of surface-bound biomolecules suggested herein can be further utilized to design effective biological scaffold for biosensor, biocatalyst, and possible diagnosis. PMID- 16378392 TI - In situ observation of interfacial bonding of an organic monolayer confined between two solid surfaces. AB - Silicon substrates coated with a long-chain hydrocarbon monolayer terminated by carboxylic acid ester groups were brought into molecular contact with different solid counter surfaces ranging from inert hydrocarbon surfaces to hydrophilic oxide surfaces. The interaction of the terminal ester groups with the counter surface was probed with infrared spectroscopy. Interfacial hydrogen bonds are reversibly formed upon contact formation, and the total degree of bonding can be adjusted by variation of the hydroxyl group density of the counter surface and quantified from the monolayer IR spectra. PMID- 16378393 TI - Cell membranes suspended across nanoaperture arrays. AB - We present a method for spreading large (>100 microm(2)) cell membrane fragments across nanoapertures in planar supports. Electron-beam and focused-ion-beam lithography were used to fabricate arrays of 50-600 nm diameter holes in free standing silicon nitride (SiN) solid films 100-500 nm thick. By pressing adhering live cells onto the nanostructured SiN surface and then removing them, planar cell membrane sheets (CMSs) were transferred in a well-defined orientation onto the SiN support. We demonstrate the accessibility to both extracellular and intracellular surfaces of CMSs by targeting membrane constituents side specifically with fluorescent markers. Our approach is of interest for studying ligand-receptor interactions using optical, electrical, and scanning probe techniques at the single-molecule level. PMID- 16378394 TI - Molecular layer-by-layer self-assembly of water-soluble perylene diimides through pi-pi and electrostatic interactions. AB - A layer-by-layer deposition process has been carried out for two oppositely charged water-soluble perylene diimide dyes without the use of intervening polyelectrolyte layers. The strong pi-pi interactions between the perylene moieties help stabilize the layers and simultaneously diminish the fluorescence quantum yield of the array without strongly affecting the absorption or fluorescence spectra. There is an alternation of fluorescence intensity according to which perylene species is on the outer layer, which is interpreted as the effect of facile energy transfer between the perylenes. PMID- 16378395 TI - Two dimensional patterning of fluorescent proteins in hydrogels. AB - This work describes the successful micropatterning of hybrid systems consisting of hydrogel-dispersed optically active and controllable proteins on solid surfaces without degradation of the photophysical properties of the light emitting biomolecules. It demonstrates the preservation of the luminescence properties of proteins entrapped into isolated microstructures of poly(acrylamide) gel. This way we can exploit both the structural and function preserving properties of the hydrogels and the functionality of light-emitting proteins. We believe that this approach can open the way to the realization of nanopatterned optical memories based on photochromic biomolecules. PMID- 16378396 TI - Tailoring surface plasmons through the morphology and assembly of metal nanoparticles. AB - Metal nanoparticles can be used as building blocks for the formation of nanostructured materials. For the design of materials with specific (optical) properties, several approaches can be followed, even when starting from the very same basic units. In this article, a survey is provided of the optical properties of noble metal nanoparticles, specifically gold, silver, and their combinations, prepared in solution through colloid chemical methods. The optical properties are shown to be mainly influenced by the surface plasmon resonance of conduction electrons, the frequency of which is not only determined by the nature of the metal but also by a number of other parameters, such as particle size and shape, the presence of a capping shell on the particle surface, or the dielectric properties of the surrounding medium. Recent results showing how these various parameters affect the optical properties are reviewed. The results highlight the high degree of control that can now be achieved through colloid chemical synthesis. PMID- 16378397 TI - Aggregation properties of cationic gemini surfactants with partially fluorinated spacers in aqueous solution. AB - The aggregation properties of cationic gemini surfactants alkanediyl-alpha,omega bis(dodecyldimethylammonium bromide), [C(12)H(25)(CH(3))(2)N(CH(2))(m)(CF(2))(n)(CH(2))(m))N(CH(3))(2)C(12)H(25)]Br(2) [where 2m + n = 12 and n = 0, 4, and 6; designated as 12-12-12, 12-12(C(4)(F)) 12, and 12-12(C(6)(F))-12, respectively] have been studied by microcalorimetry, time-resolved fluorescence quenching, and electrical conductivity. Compared with a fully hydrocarbon spacer of 12-12-12, the fluorinated spacer with a lower ratio of CF(2) to CH(2) in 12-12(C(4)(F))-12 tends to disfavor the aggregation, leading to larger critical micelle concentration (cmc), lower micelle aggregation number (N), and less negative Gibbs free energy of micellization (DeltaG(mic)). However, the fluorinated spacer with a higher ratio of CF(2) to CH(2) in 12-12(C(6)(F))-12 may prompt the aggregation, resulting in lower cmc, higher N, and more negative DeltaG(mic). It is also noted that enthalpy change of micellization (DeltaH(mic)) for 12-12(C(4)(F))-12 is the most exothermic, but the values of DeltaH(mic) for 12-12-12 and 12-12(C(6)(F))-12 are almost the same. These results are rationalized in terms of competition among the enhanced hydrophobicity and the rigidity of the fluorinated spacer, and the variation of immiscibility of the fluorinated spacer with the hydrocarbon side chains. PMID- 16378398 TI - Dynamic surface activity of phenylalanine glycerol-ether surfactant solutions measured by a differential maximum bubble pressure tensiometer. AB - A refined differential maximum bubble pressure tensiometer was used for measuring the dynamic surface tension at various concentrations of a nonconventional surfactant, a member of a new homologous series of phenylalanine glycerol-ether amphiphiles, with 10 carbon atoms to the hydrophobic alkyl chain (C(10)-PhGE). The effective bubble formation frequency for the examined surfactant concentrations was varied from 2 bubbles per second to 1 bubble per 20 s. The variation of equilibrium surface tension with concentration as well as the critical micelle concentration were determined by a Wilhelmy plate technique. Comparisons between dynamic and equilibrium surface tension values demonstrate that, under the employed surface deformation rates, the equilibrium surface tension is a misleading indicator of surface activity. This is also supported by simple surface rheology considerations. Results based on a diffusion-controlled kinetic analysis provide further evidence on the strong dependence of surface activity on the particular time scale of deformation. PMID- 16378399 TI - Do polysaccharides such as dextran and their monomers really increase the surface tension of water? AB - It has been reported in the literature that sugars such as dextrose and sucrose increase the surface tension of water. The effect was interpreted as a depletion of the solute molecules from the water-air interface. This paper presents accurate measurements of the surface tension of different concentrations of dextrose solution as well as its polymer (i.e., dextran). An automated drop shape technique called axisymmetric drop shape analysis (ADSA) was used for the surface tension determination. The surface tension measurement is presented as a function of a shape parameter, P(s), which has been used to quantify the range of the applicability of ADSA. The results of the above study show that dextrose solutions decrease the surface tension of water in contradiction to the results obtained from the weight drop method in the literature. The surface tension decreases continuously with increasing concentration. A similar effect was observed for the dextran solutions. To verify that the setup and the methodology are capable of accurately measuring increases in surface tension, a similar experiment was conducted with a sodium chloride solution with a concentration of 1 M. It is well-known that electrolyte solutions, e.g., sodium chloride, increase the surface tension of water. The results obtained from ADSA verify that the sodium chloride increases the surface tension of water by 1.6 mJ/m(2). It is concluded that dextrose and dextran decrease the surface tension of water. Thus, there is no evidence of depletion. To identify the sources of discrepancy between the results of ADSA and those reported in the literature, the experiments were repeated for different concentrations and the rate of drop formation using the drop weight method. It was found that the rate of drop formation is most likely the source of error in the results reported in the literature. PMID- 16378400 TI - Preparation of doublet, triangular, and tetrahedral colloidal clusters by controlled emulsification. AB - We describe a six-step method for making colloidal clusters of 2, 3, or 4 silica particles with a radius of 1.2 microm. This method, originally described by Manoharan et al. (Manoharan, V. N.; Elsesser, M. T.; Pine, D. J. Science 2003, 301, 483), is based on the encapsulation of silica spheres in emulsion droplets. The originality of our work lies in the preparation of monodisperse emulsions, which allows us to obtain some high yields of small aggregates over a wide range of conditions. Using optical microscopy and disk centrifugation, we show that the relative fractions of 2, 3, and 4 particle aggregates are controlled by the emulsification conditions, particularly the concentration of silica in the dispersed phase. Our best yields are obtained using low to moderate shear rates, a highly viscous continuous phase, and intermediate amounts of silica. The sedimentation of the colloidal solution into a gradient of concentration leads to aqueous suspensions of identical clusters. Since the overall process can easily be scaled up, large quantities of identical clusters may be prepared, which should allow the thermodynamic properties of these new colloidal objects to be measured for the first time. These nonspherical particles could serve as building blocks for more complex assemblies, such as colloidal crystals which could find applications as photonic materials. PMID- 16378401 TI - Experimental phase diagram of a model colloid-polymer mixture in the protein limit. AB - Sterically stabilized silica nanoparticles were synthesized, and turbidity measurements confirmed that they behaved as hard spheres in cyclohexane. Poly(isoprene) was added to give mixtures in the protein limit with a polymer coil/colloid radius ratio of 4.8. Their phase behavior under good solvent conditions was studied experimentally. The critical colloid volume fraction was phi = 0.13, whereas recent simulations (Bolhuis, P. G.; Meijer, E. J.; Louis, A. A. Phys. Rev. Lett. 2003, 90, 068304) predicted phi = 0.24. This difference is ascribed to the fact that many systems showing good solvent scaling behavior of the polymer still have a Flory-Huggins parameter close to 0.5, for instance, chi = 0.45 in this work. PMID- 16378402 TI - Structural evolution of polymer-stabilized double emulsions. AB - Polymer-stabilized double emulsions are produced by a two-step process, high shear emulsification in the primary and membrane emulsification in the secondary. By repeated fractionation after each emulsification, we obtain monodisperse double emulsions with the size of the complex droplets ranging from submicrometer to a few micrometers. With osmotic pressure balance between the inner and outer phases, the polymer-stabilized double emulsions remain stable for a year at room temperature without structure deterioration. We generalize laser light scattering to probe the structure and internal dynamics of the complex system by including the effects of the amplitude fluctuations of the scattered fields. Both static light scattering (SLS) and dynamics light scattering (DLS) can resolve the inclusions inside the complex droplets. Water-soluble nonionic surfactants are used to induce destabilization of double emulsions. We find that a double emulsion turns into a simple emulsion within a minute at a surfactant concentration of less than 10(-)(3) mol/L. We demonstrate that DLS is a powerful technique to study the kinetics of destabilization of double emulsions. Coalescence between the internal droplets and the external continuous phase is identified as a major release pathway. PMID- 16378403 TI - Layer-by-layer electrostatic self-assembly of single-wall carbon nanotube polyelectrolytes. AB - We have used anionic and cationic single-wall carbon nanotube polyelectrolytes (SWNT-PEs), prepared by the noncovalent adsorption of ionic naphthalene or pyrene derivatives on nanotube sidewalls, for the layer-by-layer self-assembly to prepare multilayers from carbon nanotubes with polycations, such as poly(diallyldimethylammonium) or poly(allylamine hydrochloride) (PDADMA or PAH, respectively), and polyanions (poly(styrenesulfonate), PSS). This is a general and powerful technique for the fabrication of thin carbon nanotube films of arbitrary composition and architecture and allows also an easy preparation of all SWNT (SWNT/SWNT) multilayers. The multilayers were characterized with vis-near-IR spectroscopy, X-ray photoelectron spectroscopy (XPS), surface plasmon resonance (SPR) measurements, atomic force microscopy (AFM), and imaging ellipsometry. The charge compensation in multilayers is mainly intrinsic, which shows the electrostatic nature of the self-assembly process. The multilayer growth is linear after the initial layers, and in SWNT/polyelectrolyte films it can be greatly accelerated by increasing the ionic strength in the SWNT solution. However, SWNT/SWNT multilayers are much more inert to the effect of added electrolyte. In SWNT/SWNT multilayers, the adsorption results in the deposition of 1-3 theoretical nanotube monolayers per adsorbed layer, whereas the nominal SWNT layer thickness is 2-3 times higher in SWNT/polyelectrolyte films prepared with added electrolyte. AFM images show that the multilayers contain a random network of nanotube bundles lying on the surface. Flexible polyelectrolytes (e.g., PDADMA, PSS) probably surround the nanotubes and bind them together. On macroscopic scale, the surface roughness of the multilayers depends on the components and increases with the film thickness. PMID- 16378404 TI - Self-depletion flocculation of tetralin oil-in-water emulsions. AB - Oil-in-water emulsions of slightly soluble oils such as tetralin prepared by high pressure homogenization and stabilized by sodium dodecyl sulfate undergo depletion flocculation induced by an initially polydisperse droplet size distribution. The smaller droplets flocculate the larger ones; the flocculation can be reversed by gentle sonication. After aging, the flocs disappear because the smaller droplets dissolve through Ostwald ripening. These effects were observed by electroacoustic measurements, supplemented by light scattering. PMID- 16378405 TI - Self-organization and pattern formation of janus particles in two dimensions by computer simulations. AB - The phase behavior and associated pattern formation of two-dimensional systems of hard disks decorated with amphiphilic coronae (Janus disks) are studied by means of Monte Carlo computer simulations. A primitive interaction potential that captures the essential interparticle interactions is introduced. Despite its simplicity, the system exhibits a very rich phase polymorphism. Apart from the isotropic phase and depending upon the coronal thickness, the simulated systems self-organize in a number of two-dimensional mesophases of various symmetries exhibiting a variety of novel patterns. The results of these simulations suggest that 2D Janus particles are promising candidates for bottom-up design of precise two-dimensional templates. PMID- 16378406 TI - Doped colloidal photonic crystal structure with refractive index chirping to the [111] crystallographic axis. AB - A three-dimensionally ordered array of close-packed colloidal spheres, a photonic crystal structure in which the refractive index of the medium interstitial lattice in a colloidal crystal spatially changes in the [111] crystallographic axis, is demonstrated. The colloidal photonic crystal structure with refractive index chirping was produced by infiltration of a monomer and organic dopants with a high refractive index into a silica opal, followed by interfacial gel polymerization. The resulting photonic crystal structure has a gradually varying stop band at each different (111) plane in the face-centered cubic (fcc) crystal structure at a normal incidence. This novel structure exhibited optical characteristics that have band-gap broadening by the superposition of stop bands at each plane of the crystal with different dielectric functions. Moreover, the refractive index perturbation in the [111] fcc opal also showed a defect state within a pseudo-photonic band gap. This new type of photonic crystal structure should be useful for the band-gap engineering of photonic-band-gap materials. PMID- 16378407 TI - Photoresponsive surfactants in microgel dispersions. AB - Microgel particles are cross-linked polymer particles. When dispersed in a good solvent for the polymer concerned, they are able to respond to a range of external stimuli by changing volume. Hence, microgel particles are suited to numerous applications (for example, controlled uptake and release) in the pharmaceutical, coatings, and water treatment industries. In this work, pH sensitive, 0.5 wt % cross-linked poly(2-vinylpyridine) (PVP) microgel particles have been prepared and characterized. When the dispersion pH is decreased below 4.5, the pyridine groups become protonated and the microgel network becomes positively charged, causing the particles to expand. To investigate the possibility of using light as a trigger for effecting volume changes, the interaction of these microgel particles with a photodegradable anionic surfactant, 4-hexylphenylazosulfonate (C(6)PAS), has been investigated using dynamic light scattering and electrophoretic mobility measurements. The electrostatic attraction between the positively charged microgel network (at solution pH 3) and the negatively charged headgroups on the surfactant molecules caused a dramatic decrease in particle volume, and charge-reversal of the particles occurred with increasing surfactant concentration. The UV irradiation of phenylazosulfonate surfactants destroys the anionic headgroup of the molecules, and the microgel particles re-swell. The irradiation of PVP dispersions in the presence of C(6)PAS, along with mixed surfactant systems of sodium dodecyl sulfate plus C(6)PAS, has been investigated. PMID- 16378408 TI - Particle-interface interaction across a nonpolar medium in relation to the production of particle-stabilized emulsions. AB - Quantitative theory of the particle-interface interaction across a nonpolar medium is developed. We consider a spherical dielectric particle (phase 1), which is immersed in a nonpolar medium (phase 2), near its boundary with a third dielectric medium (phase 3). The interaction originates from electric charges at the particle surface (e.g., the surface of a silica particle immersed in oil). The theoretical problem is solved exactly, in terms of Legendre polynomials, for arbitrary values of the dielectric constants of the three phases. As a result, expressions for calculating the interaction force and energy are derived. These expressions generalize the known theory of the electrostatic image force (acing on point charges) to the case of particles that have finite size and uniform surface charge density. For typical parameter values (silica or glass particles immersed in tetradecane), the image-force interaction becomes significant for particles of radius R > 30 nm. At fixed relative particle-to-interface distance, the force increases with the cube of the particle radius. In general, this is a strong and long-range interaction. For micrometer-sized particles, the interaction energy could be on the order of 10(5) k(B)T at close contact, and, in addition, the interaction range could be about 10(5) particle radii. The sign of the interaction depends on the difference between the dielectric constants of phases 2 and 3. When phase 3 has a smaller dielectric constant (e.g., air), the interface repels the particle. In contrast, when phase 3 has a greater dielectric constant (e.g., water), the interaction is attractive. Especially, water drops attract charged hydrophobic particles dispersed in the oily phase, and thus favor the formation of reverse particle-stabilized (Pickering) emulsions. The particle interface interaction across the oily phase is insensitive to the concentration of electrolyte in the third, aqueous phase. PMID- 16378409 TI - Nanoscale wettability of self-assembled monolayers investigated by noncontact atomic force microscopy. AB - We report on a novel technique to nucleate nanometer-sized droplets on a solid substrate and to image them with minimal perturbation by noncontact atomic force microscopy (NC-AFM). The drop size can be accurately controlled, thus permitting hysteresis measurements. We have studied the nanoscale wettability of several methyl-terminated substrates prepared by the self-assembly of organic molecules. These substrates are alkyltrichlorosilanes on silica, alkylthiols on gold, alkyl chains on hydrogen-terminated silicon, and crystalline hexatriacontane chains on silica. For each of these systems, we report a deviation of the wetting contact angle from the macroscopic value, and we discuss this effect in term of mesoscale surface heterogeneity and long-range solid-liquid interactions. PMID- 16378410 TI - Molecular bond formation between surfaces: anchoring and shearing effects. AB - Specific molecular bonds between apposing surfaces play a central role in many biological structures and functions. They display a widely varying anchoring to the cell surface, and they are subject to forces that affect their binding characteristics due to their hydrodynamic environments. Here, we examine both anchoring and shearing aspects using simplified model systems aimed at gaining insight into the formation of a 2D bond collection under stress using two different surface anchors. The highly specific streptavidin-biotin molecular bond was chosen as the model receptor-ligand pair, and grafted colloids were used as model surfaces. To explore the role of the surface anchor, we grafted biotin onto the particle surface following two different approaches: first, the grafting was performed directly on the particle amine functions; second, a 35-nm-long PEG spacer was used. Hybrid particle classes were brought into contact in a homogeneous shear (between 200 s(-)(1) and 1200 s(-)(1)) using a cone plate geometry. The bond association and dissociation kinetics were given by the time course assemblage of hybrid particles into doublets. We observed saturating kinetics profiles that we interpreted as a linkage-breakage equilibrium, which yielded the on and off rates. We found that the biotin-PEG spacer was needed in order to observe significant binding at any shear rate. We also showed that only the number of collisions per unit time, generated by the shear, affected the on rate of the binding. Neither the exerted forces nor the collision lifetime had any effect. The off rate decreased with shear, possibly because of the shortening of the force duration, which results from the increasing shear rate. PMID- 16378411 TI - Spreading of silicone oils on glass in two geometries. AB - A basic problem in liquid spreading is the hydrodynamic description of the viscous breaking force near the moving contact line. A solution to the problem of divergence at the triple line has been illustrated with two experimental configurations. It consists of observing that the rheological behavior of a Newtonian liquid is modified near the triple line due to high shear rates. Above a critical value of the shear rate, near the triple line and near the solid surface, the liquid becomes shear-thinning, meaning that the apparent viscosity of the liquid decreases as the shear rate increases. As a result, there is no divergence of the viscous energy dissipation and of the braking force as observed for a purely Newtonian behavior. This description of the viscous braking phenomenon in liquid spreading is well supported by spreading experiments of silicone oils on glass substrates in two different wetting configurations. The liquids used are two silicone oils (10,000 and 100,000 cSt). These liquids are Newtonian below a critical value of the shear rate. Above this critical value, the liquid viscosity decreases according to a power law of the shear rate. One series of experiments consider the spreading of silicone oil droplets on treated and untreated glass substrates. The other configuration consists of using the glass substrates as Wilhelmy plates and to determine the advancing dynamic contact angle as a function of the imposed speed of sinking of the plate into oil reservoirs. The two series of experiments satisfy the same dynamic wetting laws. The overall experimental results are compatible with the hypothesis of a Newtonian/non Newtonian transition of the rheological properties of liquids near the wetting front although the main origin of dissipation appears to result from Newtonian viscous braking. The same dynamic law applies for the drop and Wilhelmy plate geometries. PMID- 16378412 TI - Sorption of uranyl cations on a rutile (001) single crystal monitored by surface second-harmonic generation. AB - The rotational anisotropy of second-harmonic generation at the surface of a (001) single-crystal rutile is obtained in the presence of uranyl cations sorbed at the surface from acidic solutions at various concentrations. Surface second-harmonic generation appears to be sensitive to the presence of uranyl cations on the rutile samples. Evolution of the anisotropy pattern with initial uranyl concentration is analyzed through a phenomenological model. The elements obtained for the nonlinear susceptibility tensor Chi(2) for each sample significantly constrain the geometry of the possible sorption complexes between uranyl cations and rutile and lead to the proposition of two sorption sites involving different oxygen atoms of the rutile surface. PMID- 16378413 TI - Investigating the mechanism of chiral surface reactions: the interaction of methylacetoacetate with (S)-glutamic Acid modified Ni{111}. AB - The enantioselective hydrogenation of beta ketoesters over Ni-based catalysts is a rare example of a heterogeneously catalyzed chiral reaction. The key step in catalyst preparation is the adsorption from solution of chiral molecules (modifiers). One particularly interesting modifier is (S)-glutamic acid because the dominant enantiomeric product in the catalytic reaction depends upon the modification temperature. We report a reflection absorption infrared spectroscopy (RAIRS) study of the adsorption of methylacetoacetate (the simplest beta ketoester) onto (S)-glutamic acid modified Ni{111} surfaces as functions of the modifier coverage and modification temperature. We show that the sticking probability of methylacetoacetate is close to 0 on saturated (S)-glutamic acid covered surfaces. At lower modifier coverage, methylacetoacetate adsorption can occur. Adsorption of methylacetoacetate onto a Ni{111} surface modified by (S) glutamic acid at 300 K results in the diketo tautomeric form, with evidence being observed for a 1:1 interaction between zwitterionic (S)-glutamate and methylacetoacetate. In contrast, adsorption of methylacetoacetate onto a Ni{111} surface modified by (S)-glutamic acid at 350 K occurs exclusively in the enol tautomeric form. The implications for the heterogeneous catalytic reaction are discussed. PMID- 16378415 TI - A simple microscopic model for the dynamics of adhesive failure. AB - We consider a microscopic model for the failure of soft adhesives in tension based on ideas of bond rupture under dynamic loading. By focusing on adhesive failure under loading at constant velocity, we demonstrate that bimodal curves of stress against strain may occur due to effects of finite polymer chain or bond length and characterize the loading conditions under which such bimodal behavior is observed. The results of this analysis are in qualitative agreement with experiments performed on unconfined adhesives in which failure does not occur by cavitation. PMID- 16378414 TI - Well-defined carboxyl-terminated alkyl monolayers grafted onto H-Si(111): packing density from a combined AFM and quantitative IR study. AB - This work demonstrates that well-defined mixed carboxyl-terminated/methyl terminated alkyl monolayers can be prepared in one step on H-terminated Si(111) via direct photochemical hydrosilylation of undecylenic acid and 1-decene mixtures. As evidenced by AFM imaging and IR spectroscopy, a final rinse in hot acetic acid leaves the functionalized surface atomically smooth and perfectly free of physisorbed contaminants while unwanted material remains atop the monolayer with most other common solvents. The compositional surface chemistry was determined from a truly quantitative IR (ATR geometry) study in the range of 900-4000 cm(-)(1). Results prove that neither surface oxidation nor grafting through the carboxyl end groups occurs. Monolayers are fairly dense for such bulky end groups, with a total molecular surface density of approximately 2.7 10(14) cm(-)(2) corresponding to a surface coverage of 0.35 (maximum theoretical value approximately 0.5). Careful analysis of the CH- and COOH-related IR bands reveals that the composition of the grafted layers is richer in acid chains than the starting grafting mixture. A simple model is presented that shows that the grafting kinetics is about twice as fast for undecylenic acid as for 1-decene. Complementary electrochemical impedance measurements indicate the excellent electronic properties of the interface with a very low density of gap states. They also show that the acid terminal groups promote the penetration of water in the outer part of the organic film. PMID- 16378416 TI - Quartz crystal microbalance studies of the contact between soft, viscoelastic solids. AB - Mechanical contact between a viscoelastic lens and a viscoelastic film has been probed by means of a quartz crystal microbalance operated in the impedance analysis mode. The frequency shift induced by the formation of the contact decreases with increasing film thickness because of the finite penetration depth of the acoustic shear wave. The dependence of frequency and bandwidth on film thickness and contact area is described within a sheet-contact model, which can be employed to quantitatively analyze mechanical contact in a wide range of materials problems. The model was tested by bringing a quartz crystal coated with an elastomeric gel into contact with a hemispherical cap of a similar gel. Both gels consisted of the thermoreversible gel Kraton G swollen in mineral oil. The experiments support the model well. PMID- 16378417 TI - Preparation and structure of a low-density, flat-lying decanethiol monolayer from the densely packed, upright monolayer on gold. AB - This study investigates the formation of low-density, flat-lying decanethiol chemisorbed on Au prepared by heating the surface covered with a densely packed, upright monolayer to a surface temperature above that of the onset of desorption. We determined conditions for preparing the low-density phase by observing the evolution of the photoemission spectrum as a function of the surface temperature using polarized ultraviolet light and by utilizing scanning tunneling microscopy. The preparation conditions were similar for single- and polycrystalline gold surfaces. Once the low-density decanethiol phase was formed, reflection absorption infrared spectroscopy was employed to determine the orientation of the carbon chain backbone with respect to the Au surface. The nature of the valance electronic structure for flat-lying decanethiol is described. PMID- 16378418 TI - Polymerization of a cysteinyl peptidolipid Langmuir film. AB - The surface pressure-area isotherm of a cysteinyl peptidolipid on a pure water subphase (pH 5.8) was compared with that on a water subphase saturated with oxygen and buffered with ammonium bicarbonate (pH 7.8). A reduction of the limiting molecular area was observed for the isotherm measured on the subphase saturated with oxygen. Hysteresis in the compression-decompression cycles of the Langmuir film was also observed. Taking into consideration the chemical structure of the peptidolipid, we rationalized that the free sulfhydryl groups of the peptidolipid were oxidized in the presence of oxygen in the alkaline subphase to form intermolecular disulfide bonds at the air-water interface. The surface topography of the peptidolipid Langmuir film was observed by epi-fluorescence microscopy and the Langmuir-Blodgett film by environmental scanning electron microscopy (ESEM). The micrographs showed evidence of the polymerization of the cysteinyl peptidolipid at the air-water interface. Furthermore, the XPS spectra of the Langmuir-Blodgett films also proved the existence of disulfide bonds. The control peptidolipid C(18)-Ser-Gly-Ser-OH showed identical surface pressure-area isotherms in the presence or absence of an oxygen-saturated subphase. PMID- 16378419 TI - Collective molecular precession induced by rotating illumination in photosensitive langmuir monolayers. AB - Langmuir monolayers of an amphiphilic azobenzene derivative exhibit easily perturbable mesophases with long-range orientational order that couples strongly to linearly polarized light, resulting in symmetric photoaligned textures. Controlled rotation of the polarization plane induces continuous collective precession of the molecular field, which reveals the shear-thinning nature of the surface rotational viscosity. The dynamics is controlled by a balance between the strength of the optical coupling and viscous dissipation so that an increase in the rotational velocity results in a dampening of the synchronous anharmonic oscillations of the orientational field, as revealed by a space-time analysis of the Brewster angle microscopy images. PMID- 16378420 TI - Adsorption of simple gases in MCM-41 materials: the role of surface roughness. AB - This paper reports the development and testing of atomistic models of silica MCM 41 pores. Model A is a regular cylindrical pore having a constant section. Model B has a surface disorder that reproduces the morphological features of a pore obtained from an on-lattice simulation that mimics the synthesis process of MCM 41 materials. Both models are generated using a similar procedure, which consists of carving the pore out of an atomistic silica block. The differences between the two models are analyzed in terms of small angle neutron scattering spectra as well as adsorption isotherms and isosteric heat curves for Ar at 87 K and Xe at 195 K. As expected for capillary condensation in regular nanopores, the Ar and Xe adsorption/desorption cycles for model A exhibit a large hysteresis loop having a symmetrical shape, i.e., with parallel adsorption and desorption branches. The features of the adsorption isotherms for model B strongly depart from those observed for model A. Both the Ar and Xe adsorption branches for model B correspond to a quasicontinuous pore filling that involves coexistence within the pore of liquid bridges and gas nanobubbles. As in the case of model A, the Ar adsorption isotherm for model B exhibits a significant hysteresis loop; however, the shape of the loop is asymmetrical with a desorption branch much steeper than the adsorption branch. In contrast, the adsorption/desorption cycle for Xe in model B is quasicontinuous and quasireversible. Comparison with adsorption and neutron scattering experiments suggests that model B is too rough at the molecular scale but reproduces reasonably the surface disorder of real MCM-41 at larger length scales. In contrast, model A is smooth at small length scales in agreement with experiments but seems to be too ordered at larger length scales. PMID- 16378421 TI - Immobilization of the [RuII(edta)NO+] Ion on the surface of functionalized silica gel. AB - The reaction of NO and the immobilized dimer complex (edta)(2)Ru(2)(III(1/2),III(1/2)) on silica gel chemically modified with [3-(2 aminoethyl)aminopropyl]trimethoxysilane (AEATS) produces the corresponding immobilized nitrosyl complex AEATS/Ru(II)NO(+). This compound, a monomer, was obtained by reducing the immobilized ruthenium dimer either electrochemically or with Eu(II) and reacting this species with NO(2)(-) ions. The properties of [Ru(edta)NO](-) in solution and anchored (AEATS/Ru(II)NO(+)) on silica were compared using electrochemical (DPV, CV) and spectroscopic (IR, UV-vis, and ESR) techniques. The results indicate that immobilization does not alter the reactivity of the ruthenium complex and confirm that [Ru(edta)(H(2)O)](2)(-) may be used, either in solution or immobilized, as a catalyst for the conversion of NO(2)(-) to NO(+). Both the anchored nitrosyl complex AEATS/Ru(II)NO(+) and the [Ru(edta)NO](-) species in solution, upon one-electron reduction, liberate NO at comparable rates. PMID- 16378422 TI - Zinc(II) porphyrins at the air-water interface as studied by polarized total reflection X-ray absorption fine structure. AB - The polarized total-reflection X-ray absorption fine structure method was applied to characterize zinc porphyrins at the air-water interface. The X-ray absorption near edge structure exhibited a significant difference depending on the polarization of the X-ray. A shoulder peak of the Zn K-edge corresponding to the 1s-4p(z) transition for a square planar metal complex without axial coordination(s) was observed at 9662 eV, which indicates that the axial coordination sites of zinc porphyrin molecules examined are not fully hydrated at the air-water interface. The molecular orientation of zinc porphyrins was determined by analyzing the polarization dependence of the transition peak intensity. The meso-substituted porphyrin derivative 5,10,15,20 tetraphenylporphyrinatozinc(II) (ZnTPP) orients rather parallel to the solution surface. In contrast to ZnTPP, the zinc(II) protoporphyrin IX (ZnPP) with hydrophilic carboxyl groups at one side of the molecule stands up with respect to the solution surface, and the average tilting angle of the porphyrin plane to the surface was evaluated to be between 57 degrees and 43 degrees. In addition, the axial coordination of ZnPP is modified depending on the surface concentration, in which the axial hydration to the zinc center is effectively inhibited in the compressed surface layer. PMID- 16378423 TI - Effect of surfactants on the kinetics of nickel(II) extraction by 2-hydroxy-5 nonylacetophenone oxime (LIX 84) in an n-heptane/water system. AB - The initial reaction rates of the extraction of nickel(II) by 2-hydroxy-5 nonylacetophenone oxime (HNAPO) in a two-phase oil/water system was measured using a total internal reflectance static transfer cell. A two-step reaction mechanism between nickel(II) and HNAPO was found to satisfactorily explain the observed initial reaction rate (R(int)). The addition of neutral surfactants, nonionic octaethylene glycol mono-n-dodecyl ether and zwitterionic n dodecyldimethyl-3-ammonio-1-propanesulfonate, decreased R(int), which could be accounted for with a competitive surface adsorption model. The presence of the anionic surfactant sodium dodecyl sulfate accelerated and then decelerated R(int), while the cationic surfactant dodecyltrimethylammonium chloride caused a decrease. The effects of these charged surfactants were accounted for using a combination of a competitive surface adsorption model and the Boltzmann distribution of charged species. PMID- 16378424 TI - Mechanical model for anisotropic curved interfaces with applications to surfactant-laden liquid-liquid crystal interfaces. AB - A mechanical model for anisotropic curved interfaces, applicable to thermodynamically closed surfactant-laden liquid-liquid crystal interfaces is developed. The model takes into account the mechanical effects due to surface bending and surface tilting (anchoring) and incorporates liquid crystal anisotropy into classical fluid membrane mechanics. In the absence of the aligned liquid crystal, the model converges to the fluid membrane mechanical model, and in the absence of surfactant, it converges to the nematic interface mechanical model. Use of the well-known Helfrich-Rapini-Papoular surface energies leads to the Laplace equation for anisotropic curved interfaces, whose material limits are the vesicle shape equation and the liquid crystal Herring equation. Applications of the model to shape selection in liquid drops embedded in aligned nematic liquid crystals illustrates the competition between anchoring and bending and shows how anisotropic surface tension distorts the droplet and how bending tends to restore the spherical shape. This theory presented in this article shows that the interaction of interfacial anchoring and bending creates new regimes in classical fluid membrane mechanics. PMID- 16378425 TI - Measurements of interfacial viscoelasticity with a quartz crystal microbalance: influence of acoustic scattering from a small crystal-sample contact. AB - We discuss the influence of a limited contact size on measurements of high frequency interfacial viscoelasticity performed with a combination of a quartz crystal microbalance (QCM) and the Johnson-Kendall-Roberts (JKR) apparatus. In this instrument, a sphere-plate contact is established between an elastomeric lens and a quartz resonator. The analysis is carried out in the frame of the sheet-contact model, which states that both the shift of resonance frequency and the bandwidth are proportional to the contact area as long as the contact area is much smaller than the crystal itself. In particular, the ratio of the shift in bandwidth and the shift in frequency (termed the D-f ratio) is predicted to be constant and independent of geometry. However, the experiment does show a slight increase in the D-f ratio with the contact radius when the contact radius is comparable to the wavelength of sound inside the crystal. This effect can be explained by acoustic scattering. PMID- 16378426 TI - Adsorption of xenon on purified HiPco single walled carbon nanotubes. AB - We have measured adsorption of xenon on purified HiPco single-walled carbon nanotubes (SWNTs) for coverages in the first layer. We compare the results on this substrate to those our group obtained in earlier measurements on lower purity arc-discharge produced nanotubes. To obtain an estimate for the binding energy of Xe, we measured five low-coverage isotherms for temperatures between 220 and 260 K. We determined a value of 256 meV for the binding energy; this value is 9% lower than the value we found for arc discharge nanotubes and is 1.59 times the value found for this quantity on planar graphite. We have measured five full monolayer isotherms between 150 and 175 K. We have used these data to obtain the coverage dependence of the isosteric heat. The experimental values obtained are compared with previously published computer simulation results for this quantity. PMID- 16378428 TI - Interfacial properties of a synthetic peptide derived from hepatitis G virus E2 protein: interaction with lipid monolayers. AB - A useful approach to get information about the potential fusogenic ability of virus synthetic peptides is the study of its interfacial properties and subsequent study in mono- and bilayers. In this work, we have characterized by means of physicochemical tools (i.e. compression isotherms and surface activity) the sequence 267-284, LLGTEVSEVLGGAGLTGG, derived from the E2 structural protein of HGV/GBV-C. The adsorption of the peptide at the air/water interface was monitored by following the increase in surface pressure as a function of time at two different pH values: 5 and 7. Parameters such as surface excess or molecular area were calculated from the equation of Gibbs. The peptide showed a tendency to migrate to the surface of a saline-buffered solution. It formed stable monolayers at the air/water interface giving a compression isotherm with a shape consistent with that of some alpha-helical peptide conformations. Brewster angle microscopy (BAM) showed that through compression the peptide formed multilayers. The studies with lipid monolayers (DPMC, DMPC/DMPG, and DMPC/DMTAP) showed that the peptide interacts with all the lipids assayed producing a marked disrupting effect upon them. In these effects electrostatic interactions seem to have some participation. PMID- 16378427 TI - Structure/property relationships in erodible multilayered films: influence of polycation structure on erosion profiles and the release of anionic polyelectrolytes. AB - We have investigated the influence of polymer structure on the erosion profiles of multilayered polyelectrolyte assemblies fabricated from sodium poly(styrene sulfonate) (SPS) and three different hydrolytically degradable polyamines. We synthesized three structurally related poly(beta-amino ester)s (polymers 1-3) having systematic variations in both charge density and hydrophobicity. These changes in structure did not influence film thickness significantly, but polymer structure was found to play an important role in defining the rates at which multilayered assemblies fabricated from these materials eroded in physiologically relevant media. Films 60 nm thick fabricated from polymer 1 and SPS eroded completely in 50 h when incubated in PBS buffer at 37 degrees C, as determined by ellipsometry. Analogous films fabricated from polymers 2 and 3 eroded and released SPS into solution over significantly longer time periods ranging from approximately 150 h (ca. 6 days) to 370 h (ca. 15 days), respectively. These differences are consistent with a systematic increase in the hydrophobicity of polymers 1-3 as well as the relative rates at which these polymers degrade hydrolytically. This work demonstrates that it is possible to tailor the rates at which thin, multilayered polyelectrolyte assemblies release incorporated anionic polyelectrolytes over a large range of time periods simply by changing the structure of the degradable polyamine used to fabricate a film. The principles reported here may therefore contribute to the design of multilayered assemblies that permit a broad range of spatial and temporal control over the release of therapeutic agents from coated surfaces. PMID- 16378429 TI - Combination of electrografting and atom-transfer radical polymerization for making the stainless steel surface antibacterial and protein antiadhesive. AB - A two-step "grafting from" method has been successfully carried out, which is based on the electrografting of polyacrylate chains containing an initiator for the atom transfer radical polymerization (ATRP) of 2-(tert-butylamino)ethyl methacrylate (TBAEMA) or copolymerization of TBAEMA with either monomethyl ether of poly(ethylene oxide) methacrylate (PEOMA) or acrylic acid (AA) or styrene. The chemisorption of this type of polymer brushes onto stainless steel surfaces has potential in orthopaedic surgery. These films have been characterized by ATR FTIR, Raman spectroscopy, atomic force microscopy (AFM), and measurement of contact angles of water. The polymer formed in solution by ATRP and that one detached on purpose from the surface have been analyzed by size exclusion chromathography (SEC) and (1)H NMR spectroscopy. The strong adherence of the films onto stainless steel has been assessed by peeling tests. AFM analysis has shown that addition of hydrophilic comonomers to the grafted chains decreases the surface roughness. According to dynamic quartz crystal microbalance experiments, proteins (e.g., fibrinogen) are more effectively repelled whenever copolymer brushes contain neutral hydrophilic (PEOMA) co-units rather than negatively charged groups (PAA salt). Moreover, a 2- to 3-fold decrease in the fibrinogen adsorption is observed when TBAEMA is copolymerized with either PEOMA or AA rather than homopolymerized or copolymerized with styrene. Compared to the bare stainless steel surface, brushes of polyTBAEMA, poly(TBAEMA-co-PEOMA) and poly(TBAEMA-co-AA) decrease the bacteria adhesion by 3 to 4 orders of magnitude as revealed by Gram-positive bacteria S. aureus adhesion tests. PMID- 16378430 TI - Chemomechanics of surface stresses induced by DNA hybridization. AB - When biomolecular reactions occur on one surface of a microcantilever beam, changes in intermolecular forces create surface stresses that bend the cantilever. While this phenomenon has been exploited to create label-free biosensors and biomolecular actuators, the mechanisms through which chemical free energy is transduced to mechanical work in such hybrid systems are not fully understood. To gain insight into these mechanisms, we use DNA hybridization as a model reaction system. We first show that the surface grafting density of single stranded probe DNA (sspDNA) on a surface is strongly correlated to its radius of gyration in solution, which in turn depends on its persistence length and the DNA chain length. Since the persistence length depends on ionic strength, the grafting density of sspDNA can be controlled by changing a solution's ionic strength. The surface stresses produced by the reaction of complementary single stranded target DNA (sstDNA) to sspDNA depend on the length of DNA, the grafting density, and the hybridization efficiency. We, however, observe a remarkable trend: regardless of the length and grafting density of sspDNA, the surface stress follows an exponential scaling relation with the density of hybridized sspDNA. PMID- 16378431 TI - On the kinetics of apatite growth on substrates under physiological conditions. AB - Derived from reaction kinetics, a simple but useful method, based on "apatite forming capacity" or AFC of solutions mimicking blood plasma, is proposed to decipher the rate of calcium phosphate mineralization. Apatite growth rate constants were calculated using this method for a model composite surface varying the volume fraction of synthetic hydroxyapatite (HA) in a polymer matrix. Previously reported data for mineralized surfaces on Ta, Ti, and its alloys are also analyzed similarly and compared. Utilizing the growth rate constant, the bioactivity of the materials was indexed in vitro. Complementarily, semiempirical quantum mechanical calculation (ZINDO method) showed that the interaction of cations with TRIS-hydroxymethyl aminomethane molecules in the solution is stronger than that with the polymer substrate considered, but weaker than hydrated Ti and TiO(2) surfaces. This analysis then quantifies for example the extent of polymer inertness and the "bioactivity" of alkali treated Ti. The growth rate constants for the model materials prepared in this work are explained on the basis of localized dissolution of HA, the amount of which simply increases with increasing volume fraction of HA in the composite. PMID- 16378432 TI - Glass surfaces grafted with high-density poly(ethylene glycol) as substrates for DNA oligonucleotide microarrays. AB - Surfaces carrying a dense layer of poly(ethylene glycol) (PEG) were prepared, characterized, and tested as substrates for DNA oligonucleotide microarrays. PEG bis(amine) with a molecular weight of 2000 was grafted onto silanized glass slides bearing aldehyde groups. After grafting, the terminal amino groups of the PEG layer were derivatized with the heterobifunctional cross-linker succinimidyl 4-[p-maleimidophenyl]butyrate to permit the immobilization of thiol-modified DNA oligonucleotides. The stepwise chemical modification was validated with X-ray photoelectron spectroscopy. Goniometry indicated that the PEG grafting procedure reduced surface inhomogeneities present after the silanization step, while atomic force microscopy and ellipsometry confirmed that the PEG layer was dense and monomolecular. Hybridization assays using DNA oligonucleotides and fluorescence imaging showed that PEG grafting improved the yield in hybridization 4-fold compared to non-PEGylated maleimide-derivatized surfaces. In addition, the PEG layer reduced the nonspecific adsorption of DNA by a factor of up to 13, demonstrating that surfaces with a dense PEG layer represent suitable substrates for DNA oligonucleotide microarrays. PMID- 16378433 TI - Monitoring metal ion binding in single-layer Pseudomonas aeruginosa biofilms using ATR-IR spectroscopy. AB - The binding of metal ions to Pseudomonas aeruginosa PAO1 cells attached to a ZnSe surface has been observed in this research through cation exchange experiments using ATR-IR spectroscopy. A biofilm consisting of a single layer of Pseudomonas aeruginosa PAO1 cells was formed on a ZnSe prism by flowing a bacterial suspension in a 0.03 mol L(-)(1) NaNO(3) solution at pH 5.0 across its surface. Exposure of the biofilm to chromium(III) nitrate solution resulted in increases in all band absorbances. This absorbance increase has been attributed to the binding of chromium(III) to the bacterial exopolymers associated with the prism surface. The chromium(III) binding causes the exopolymers to contract and move the bacterial cell closer to the ZnSe surface. Further study of chromium(III) ion exchange using a mutant P. aeruginosa with a truncated lipopolysaccharide (LPS) chain resulted in much smaller absorbance changes. This observation supports the view that the extension of bacterial exopolymers and hence the distance of the bacterial cell from the surface is strongly influenced by environmental factors such as the presence of metal cations. Following chromium(III) cation exchange, the bacterial band absorbances remained constant even when the bacteria were washed with a 0.03 mol L(-)(1) NaNO(3) solution, indicating that the chromium(III) was irreversibly bound. Ion exchange with nickel(II) and cobalt(II) nitrate solutions within identical biofilms showed that these cations caused relatively small increases in absorbances that were reversible, indicating that nickel(II) and cobalt(II) are less strongly bound than chromium(III) within P. aeruginosa biofilms. The absence of discernible IR spectral changes with metal binding appears to indicate a predominantly electrostatic mechanism for binding of Cr(III), Ni(II), and Co(II) ions by bacteria in the early stages of biofilm formation. PMID- 16378434 TI - Organized arrays of individual DNA molecules tethered to supported lipid bilayers. AB - An unappreciated aspect of many single-molecule techniques is the need for an inert surface to which individual molecules can be anchored without compromising their biological integrity. Here, we present new methods for tethering large DNA molecules to the surface of a microfluidic sample chamber that has been rendered inert by the deposition of a supported lipid bilayer. These methods take advantage of the "bio-friendly" environment provided by zwitterionic lipids, but still allow the DNA molecules to be anchored at fixed positions on the surface. We also demonstrate a new method for constructing parallel arrays of individual DNA molecules assembled at defined positions on a bilayer-coated, fused silica surface. By using total internal reflection fluorescence microscopy to visualize the arrays, it is possible to simultaneously monitor hundreds of aligned DNA molecules within a single field-of-view. These molecular arrays will significantly increase the throughput capacity of single-molecule, fluorescence based detection methods by allowing parallel processing of multiple individual reaction trajectories. PMID- 16378436 TI - SANS analysis of the microstructural evolution during the aging of pyrolysis oils from biomass. AB - In this paper, we report for the first time a microstructural characterization of pyrolysis oils obtained from biomass. Bio crude oils (BCOs) are good candidates as substitutes for mineral oils as fuels. By using small-angle neutron scattering (SANS), we show that BCOs are nanostructured fluids constituted by a complex continuous phase and nanoparticles mainly formed by the association of units of pyrolytic lignins. The aggregation of these units during the time produces branched structures with fractal dimension D(f) between 1.4 and 1.5, which are responsible for BCO aging. SANS results fully support the recently formulated thermal ejection theory, accounting for the mechanism of formation of the lignin fraction in oils obtained from fast pyrolysis of biomass. PMID- 16378435 TI - Gold nanoparticles as carriers for efficient transmucosal insulin delivery. AB - Nanomaterials have gained tremendous importance in biology and medicine because they can be used as carriers for delivering small molecules such as drugs, proteins, and genes. We report herein the binding of the hormone insulin to gold nanoparticles and its application in transmucosal delivery for the therapeutic treatment of diabetes mellitus. Insulin was loaded onto bare gold nanoparticles and aspartic acid-capped gold nanoparticles and delivered in diabetic Wistar rats by both oral and intranasal (transmucosal) routes. Our principle observations are that there is a significant reduction of blood glucose levels (postprandial hyperglycemia) when insulin is delivered using gold nanoparticles as carriers by the transmucosal route in diabetic rats. Furthermore, control of postprandial hyperglycemia by the intranasal delivery protocol is comparable to that achieved using the standard subcutaneous administration used for type I diabetes mellitus, thus showing considerable promise for further development. PMID- 16378437 TI - Colloid probe AFM investigation of interactions between fibrinogen and PEG-like plasma polymer surfaces. AB - Interaction forces between surfaces designed to be protein resistant and fibrinogen (Fg) were investigated in phosphate-buffered saline with colloid probe atomic force microscopy. The surfaces of the silica probes were coated with a layer of fibrinogen molecules by adsorption from the buffer. The technique of low power, pulsed AC plasma polymerization was used to make poly(ethylene glycol) (PEG)-like coatings on poly(ethylene teraphthalate) by using diethylene glycol vinyl ether as the monomer gas. The degree of PEG-like nature of the films was controlled by use of a different effective plasma power in the chamber for each coating, ranging from 0.6 to 3.6 W. This produced a series of thin films with a different number of ether carbons, as assessed by X-ray photoelectron spectroscopy. The interaction force measurements are discussed in relation to trends observed in the reduction of fibrinogen adsorption, as determined quantitatively by (125)I radio-labeling. The plasma polymer coatings with the greatest protein-repelling properties were the most PEG-like in nature and showed the strongest repulsion in interaction force measurements with the fibrinogen coated probe. Once forced into contact, all the surfaces showed increased adhesion with the protein layer on the probe, and the strength and extension length of adhesion was dependent on both the applied load and the plasma polymer surface chemistry. When the medium was changed from buffer to water, the adhesion after contact was eliminated and only appeared at much higher loads. This indicates that the structure of the fibrinogen molecules on the probe is changed from an extended conformation in buffer to a flat conformation in water, with the former state allowing for stronger interaction with the polymer chains on the surface. These experiments underline the utility of aqueous surface force measurements toward understanding protein-surface interactions, and developing nonfouling surfaces that confer a steric barrier against protein adsorption. PMID- 16378438 TI - Selective betainization of PS-P4VP and solution properties. AB - The ability to control the structure and function of supramolecular self assemblies gives rise to many patterning possibilities. Making use of the interactions between the copolymers and solvent provides a neat way of controlling the structures. The pyridine residues of each of the diblock copolymer polystyrene-b-poly(4-vinylpyridine) (PS-P4VP) was selectively betainized using 1,3-propane sultone under mild conditions to yield a series of novel betaine diblock copolymers in chloroform and toluene, respectively. The solution properties were studied using dynamic light scattering, static light scattering, and (1)H NMR spectroscopy. The morphology of the resulting micellar film was studied using transmission electron microscopy. The size of the micelles formed was found to be strongly dependent on the amount of sultone grafted and the shape of PS-P4VP-sultone micelle changed from spherical to elongated with different PS-P4VP-to-sultone ratio. PMID- 16378439 TI - Silica monoliths templated on L3 liquid crystal. AB - Dimensionally stable, optically clear, highly porous (approximately 65% of the apparent volume), and high surface area (up to 1400 m(2)/g) silica monoliths were fabricated as thick disks (0.5 cm) by templating the isotropic liquid crystalline L(3) phase with silica through the hydrolysis and condensation of a silicon alkoxide and then removing the organic constituents by supercritical ethanol extraction. The L(3) liquid crystal is a stable phase formed by the cosurfactants cetylpyridinium chloride monohydrate and hexanol in HCl(aq) solvent. Extracted 0.5 cm thick disks exhibited a low ratio of scattered to transmitted visible light (1.5 x 10(-)(6) at 22 degrees from the surface normal). The degree of silica condensation in the monoliths was high, as determined by (29)Si NMR measurements of Q(3) and Q(4) peak intensities (0.53 and 0.47, respectively). As a result, the extracted and dried monoliths were mechanically robust and did not fracture when infiltrated by organic solvents. Photoactive liquid monomers were infiltrated into extracted silica monoliths and polymerized in situ, demonstrating the possible application of templated silica to optical storage technology. PMID- 16378440 TI - Micropatterning of copper on a poly(ethylene terephthalate) substrate modified with a self-assembled monolayer. AB - We have developed a technique for the site-selective electroless deposition of Cu on poly(ethylene terephthalate) (PET) substrate modified with an organic self assembled monolayer (SAM). The PET substrate was first modified with a silica like layer by being dip-coated in an acetone solution of 3 aminopropyltrimethoxysilane and treated with UV light. The PET substrate was further modified with thiol groups using a 3-mercaptopropyltrimethoxysilane-SAM and then irradiated by UV light through a photomask to prepare thiol-group regions and OH-group regions. Cu was then deposited on only the thiol-group regions of the substrate by electroless deposition in a neutral solution with no catalysts by using dimethylamineborane as a reducing reagent. This site-selective deposition process can control the deposition conditions by an organic thin film fabricated on a surface-modified PET substrate, and thus can be applied to other low heat-resistant substrates. PMID- 16378441 TI - Composite thin film by hydrogen-bonding assembly of polymer brush and poly(vinylpyrrolidone). AB - Based on hydrogen-bonding layer-by-layer (LBL) assembly in aqueous solution, poly(vinylpyrrolidone) (PVPON) and a spherical polymer brush with a poly(methylsilsesquioxane) (PSQ) core and poly(acrylic acid) (PAA) hair chains were used to fabricate composite multilayer thin films. Hydrogen bonding as the driving force was confirmed by FT-IR spectrometry. A simple method (Filmetric F20) was introduced to determine the thickness and refractive index of the films. The film thickness was found to be a linear function of the number of bilayers. The average increase in thickness per bilayer is 28.3 nm. The film morphology was characterized with scanning electron microscopy and atomic force microscopy. The images obtained from the two instruments show a great resemblance. The films were further calcined to get an inorganic film by removing the organic components, or treated with tetrabutylammonium fluoride (TBAF) to remove the PSQ core and get an organic film. The optical properties and morphological changes induced by these treatments were also studied. PMID- 16378442 TI - Solvent-stabilized oxovanadium phthalocyanine nanoparticles and their application in xerographic photoreceptors. AB - A stable organic sol of solvent-stabilized oxovanadium phthalocynine (VOPc) nanoparticles with excellent photoconductivity was successfully prepared by ultrasonificating a prepared nanoscopic VOPc powder in1,2-dichloroethane (C(2)H(4)Cl(2)) without any additive. These solvent-stabilized VOPc nanoparticles have a size distribution from 2 to 20 nm with an average diameter of 4.6 nm. The VOPc concentration of these organic sols could be as high as 100 g/L. The nanoscopic VOPc particles were well-dispersed in an insulating polycarbonate (PC) resin, resulting in single-layered photoreceptors with high surface charge durability in the dark and excellent photoconductivity. Based on the light assisted scanning tunneling microscopy (STM) measurements, the charge transport mechanism of these photoreceptors was ascribed to light-induced enhancement of electron tunneling through the VOPc-nanoparticle/insulator junctions. PMID- 16378443 TI - Liesegang pattern formation in kappa-carrageenan gel. AB - We report a new class of the spatial pattern formation process in which the gel plays essential roles. The system studied here is the solution of kappa carrageenan in which potassium chloride is diffused. The solution transforms into the gel state with the diffusion of potassium chloride. Then the stripe pattern, which is perpendicular to the direction of the diffusion of potassium chloride, appears within the gel. The pattern thus formed in the gel is studied as a function of the concentration of the solution of potassium chloride. We find that the dense region of the stripe pattern consists of the liquid crystalline gel, whereas the dilute region is the amorphous gel. The transition from the amorphous gel to the liquid crystalline gel, hence, occurs in the gel state of kappa carrageenan. The gel behaves as a pattern-forming substance as well as the supporting medium of the pattern in this system. The period and the thickness of the layers of liquid crystalline gel are analyzed. Both the period and the thickness of the layers are found to depend strongly on the concentration of the solution of potassium chloride. PMID- 16378444 TI - Composition-tunable properties of amphiphilic comb copolymers containing protected methacrylic acid groups for multicomponent protein patterning. AB - Methods to micropattern multiple protein components on surfaces under mild conditions are of interest for biosensing, proteomics, and fundamental studies in cell biology. Here, we report on the composition-dependent thin-film solubility behavior of o-nitrobenzyl methacrylate (oNBMA, a protected form of methacrylic acid)/methyl methacrylate (MMA)/poly(ethylene glycol) methacrylate (PEGMA) random terpolymers, materials which are promising as aqueous-processible photoresists. Over a broad range of terpolymer compositions, these materials formed initially water-insoluble films, which, upon UV irradiation, rapidly dissolved in aqueous solutions above a critical pH. This threshold pH ranged from approximately 5-7 depending upon the copolymer composition and decreased as the relative ratio of MMA to PEGMA in the copolymers decreased. In addition, in a narrow window of compositions near 35:0:65 oNBMA/MMA/PEGMA (wt ratio), an inverse behavior was observed: thin films that were initially water soluble became kinetically stable in aqueous solutions after UV exposure. The time for these films to completely dissolve was hours rather than seconds, and the rate of dissolution was both temperature- and pH-dependent. This behavior is consistent with a transient stability imparted by inter- and intramolecular hydrogen bonding in the film. Using copolymers of this composition as negative tone photoresists, we demonstrated patterning of two proteins into two discrete regions of a surface. The selective solubility of the resist copolymer allows the entire patterning process to be completed using only biological buffers as solvents and across a temperature range between 4 and 37 degrees C without subjecting either protein to ultraviolet irradiation or dehydration. These materials are thus of interest for complex surface photopatterning under mild aqueous conditions. PMID- 16378445 TI - Fabrication of fluorescent nanotubes based on layer-by-layer assembly via covalent bond. AB - A pressure-filter-template approach was employed to prepare fluorescent nanotubes of polyethyleneimine (PEI) and 3,4,9,10-perylenetetracarboxylicdianhydride (PTCDA) through covalent combination in the porous of alumina template based on the layer-by-layer (LbL) assembly technique. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) images exhibited a tubular structure of the obtained samples. The wall thickness of the nanotubes is well controlled by varying the assembled cycle number, which is detected by UV-visible spectroscopy. Fourier transform infrared (FT-IR) spectroscopy confirmed the formation of covalent bonds between PEI and PTCDA in nanotubes. PMID- 16378446 TI - AFM study of defect-induced depressions of the smectic-A/air interface. AB - The smectic-A/air interface of liquid-crystal droplets with antagonistic boundary conditions is studied by atomic force microscopy (AFM). The droplets are prepared on coated silicon wafers on which a planar alignment is preferred in contrast to the homeotropic alignment at the air interface. As a result, focal conic defects appear in the smectic-A phase causing a characteristic pattern of depressions in the droplet surface. The dimensions of the defect-induced depressions are measured by AFM as a function of temperature for two different compounds possessing a smectic-A-isotropic and a smectic-A-nematic transition. Whereas the results are independent of temperature in the smectic-A-isotropic case, reflecting the first-order nature of the transition, a pronounced temperature dependence is observed for the second compound, where the depth of the defect induced depressions decreases continuously with increasing temperature and vanishes at the second-order transition to the nematic phase. These observations can be qualitatively explained through the behavior of the layer compressional elastic constant at the smectic-A-nematic transition. PMID- 16378447 TI - Mechanism of smectic arrangement of montmorillonite and bentonite clay platelets incorporated in gels of poly(acrylamide) induced by the interaction with cationic surfactants. AB - Structure transitions, induced by the interaction with the cationic surfactant cetylpyridinium chloride in nanocomposite gels of poly(acrylamide) with incorporated suspensions of the two closely related layered clays bentonite and montmorillonite, were studied. Unexpectedly, different behaviors were revealed. X ray diffraction measurements confirm that, due to the interaction with the surfactant, initially disordered bentonite platelets arrange into highly ordered structures incorporating alternating clay platelets and surfactant bilayers. The formation of these smectic structures also in the cross-linked polymer gels, upon addition of the surfactant, is explained by the existence of preformed, poorly ordered aggregates of the clay platelets in the suspensions before the gel formation. In the case of montmorillonite, smectic ordering of the disordered platelets in the presence of the surfactant is observed only after drying the suspensions and the clay-gel composites. Rheology studies of aqueous suspensions of the two clays, in the absence of both surfactant and gel, evidence a much higher viscosity for bentonite than for montmorillonite, suggesting smaller clay aggregate size in the latter case. Qualitatively consistent results are obtained from optical micrographs. PMID- 16378448 TI - Radical-induced generation of small silver particles in SPEEK/PVA polymer films and solutions: UV-Vis, EPR, and FT-IR studies. AB - The present study is centered on the processes involved in the photochemical generation of nanometer-sized Ag particles via illumination at 350 nm of aqueous solutions and cross linked films containing sulfonated poly(ether ether ketone) and poly(vinyl alcohol). Optical and electron paramagnetic resonance experiments, including electron nuclear double resonance data, proved conclusively that the photogenerated chromophore exhibiting a band with lambda(max) = 565 nm is an alpha-hydroxy aromatic (ketyl) radical of the polymeric ketone. This reducing species was produced by illumination of either solutions or films, but the radical lifetime extended from minutes in the fluid phase to hours in the solid. Direct evidence is presented that this long-lived chromophore reduces Ag(I), Cu(II), and Au(III) ions in solution. A rate constant of k = 1.4 x 10(3) M(-)(1) s(-)(1) was obtained for the reduction of Ag(+) by the ketyl radical from the post-irradiation formation of Ag crystallites. FTIR results confirmed that the photoprocess yielding polymeric ketyl radicals involves a reaction between the macromolecules. The photochemical oxidation of the polymeric alcohol, as well as the formation of light-absorbing macromolecular products and polyols, indicates that the sulfonated polyketone experienced transformations similar to those encountered during illumination of the benzophenone/2-propanol system. PMID- 16378449 TI - Cobalt sorption in silica-pillared clays. AB - Silicon pillared samples were prepared following conventional and microwave irradiation methods. The samples were characterized and tested in cobalt sorption. Ethylenediammine was added before cobalt addition to improve the amount of cobalt retained. The amount of cobalt introduced in the original clay in the presence of ethylenediammine was the highest. In calcined pillared clays the cobalt retention with ethylenediammine was lower (ca. 40%). In all cases the presence of ethylenediammine increased twice the amount of cobalt sorption measured for aqueous solutions. PMID- 16378450 TI - Adsorption free energy of variable-charge nanoparticles to a charged surface in relation to the change of the average chemical state of the particles. AB - Variable-charge nanoparticles such as proteins and humics can adsorb strongly to charged macroscopic surfaces such as silica and iron oxide minerals. To model the adsorption of variable-charge particles to charged surfaces, one has to be able to calculate the adsorption free energy involved. It has been shown that the change in the free energy of variable-charge particles is related to the change in their average chemical state upon adsorption, which is commonly described using surface complexation models. In this work, expressions for the free-energy change in variable-charge particles due to changes in chemical binding are derived for three ion-binding models (i.e., the Langmuir, Langmuir-Freundlich, and NICA models) and for changes due to nonspecific binding for the Donnan model. The expressions for the adsorption free energy of the variable-charge particles to a charged surface are derived on the basis of the equality of the (electro)chemical potential of the particles in the bulk solution and adsorption phase. The expressions derived are general in the sense that they account for the competition between charge-determining ions that bind chemically to the particles, and they also apply in case of the formation of chemical bonds between particle ligands and surface sites. The derived expressions can be applied in the future to model the adsorption of variable-charge nanoparticles to charged surfaces. The results obtained for the NICA-Donnan model make it possible to apply this advanced surface complexation model to describe the adsorption of humics to minerals. PMID- 16378451 TI - Sonochemical synthesis of hollow PbS nanospheres. AB - PbS hollow nanospheres with diameters of 80-250 nm have been synthesized by a surfactant-assisted sonochemical route. The nanostructures were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), (high resolution) transmission electron microscopy [(HR)TEM], and scanning electron microscopy (SEM) images. Structural characterization indicates that shells of the hollow spheres are composed of PbS nanoparticles with diameters of about 12 nm. The formation of the hollow nanostructure was explained by a vesicle-template mechanism, in which sonication and surfactant play important roles. Furthermore, uniform silica layers were successfully coated onto the hollow spheres via a modified Stober method to enhance their performance for promising applications. PMID- 16378452 TI - Two-step growth of goethite from ferrihydrite. AB - Goethite (alpha-FeOOH) is an antiferromagnetic iron oxyhydroxide that is often synthesized by precipitation from homogeneous, aqueous solution followed by aging. This paper addresses goethite growth by phase transformation of six-line ferrihydrite nanoparticles to goethite followed by oriented aggregation of the goethite primary particles. Data tracking goethite nanocrystal growth as a function of pH, temperature, and time is presented. In general, goethite growth by oriented aggregation is faster at higher pH and at higher temperature even as growth by coarsening becomes increasingly important as pH increases. In addition, particle size measurements demonstrate that the primary nanoparticles grow by Ostwald ripening even as they are being consumed by oriented aggregation. Finally, the use of a microwave anneal step in the preparation of the precursor six-line ferrihydrite nanoparticles substantially improves the homogeneity of the final goethite product. Final goethite nanoparticles are unaggregated, acicular crystals in the tens of nanometers size range. These particles may be ideal for mineral liquid crystal and magnetic-recording media applications. PMID- 16378453 TI - Pore anisotropy and microporosity in nanostructured mesoporous solids. AB - In this study, we carried out an investigation related to the determination of the anisotropy (b) of pores as well as the extent of microporosity (mic%) in various groups of nanostructured mesoporous materials. The mesoporous materials examined were fifteen samples belonging to the following groups of solids: MCM 48s, SBA-15s, SBA-16s, and mesoporous TiO(2) anatases. The porosities of those materials were modified either during preparation or afterward by the addition of Cu(II) species and/or 3(5)-(2-pyridinyl) pyrazole (PyPzH) into the pores. The modification of porosity in each group took place to make possible the internal comparison of the b and mic% values within each group. The estimation of both the b and mic% parameters took place from the corresponding nitrogen adsorption desorption isotherms. The new proposed method is able to detect a percentage of microporosity as low as a few percent, which is impossible by any of the methods used currently, without the use of any reference sample or standard isotherms. A meaningful inverse relationship is apparent between the b and mic% values, indicating that large values of b correspond to small values of mic%. PMID- 16378454 TI - Self-assembly of mixtures of block copolymers of poly(styrene-b-acrylic acid) with random copolymers of poly(styrene-co-methacrylic acid). AB - The effects of the addition of random copolymers of poly(styrene-co-methacrylic acid) [P(S-co-MAA)] on the self-assembly of block copolymers of poly(styrene-b acrylic acid) (PS-b-PAA) are described. The effects of variation of five factors, including the MAA content, the weight fraction and molar mass of the P(S-co-MAA), the initial concentration of the mixture, and the length of the PAA segment in the block copolymer, were investigated. With increasing MAA content, the localization of the random copolymer in the aggregate changed from the core to the interface, which led to a morphological transition from spheres to vesicles. Vesicles, mixtures of vesicles and large spheres, and large spheres alone were formed with increasing weight fraction of the random copolymer. When the molar mass of the random copolymer was high, both rods and vesicles were observed at low water contents; otherwise, only vesicles were observed. The vesicle size increased (from 100 to 140 nm) with increasing initial polymer concentration, whereas the vesicle membrane thickness remained constant. The size of the vesicles prepared from the mixtures increased with water content but decreased with the length of PAA in the diblock. PMID- 16378455 TI - Thermal modulated interaction of aqueous steroids using polymer-grafted capillaries. AB - Poly(N-isopropylacrylamide) (PIPAAm) of controlled molecular weight was densely grafted onto glass capillary lumenal surfaces using surface-initiated atom transfer radical polymerization (ATRP). Temperature-dependent changes of these thermoresponsive brush surfaces with hydrophobic steroids were investigated by exploiting thermoresponsive aqueous wettability changes of the polymer-modified surfaces in microfluidic systems. IPAAm was polymerized on ATRP initiator immobilized glass surfaces using CuCl/CuCl(2)/tris(dimethylaminoethyl)amine (Me(6)TREN) as an ATRP catalyst in water at 25 degrees C. PIPAAm graft layer thickness and its homogeneity on glass surfaces are controlled by changing ATRP reaction time. Aqueous wettability changes of PIPAAm-grafted surfaces responses drastically changed to both grafted polymer layer thickness and temperature, especially at lower temperatures. Temperature-responsive surface properties of these PIPAAm brushes within capillary inner wall surfaces were then investigated using capillary chromatography. Effective interaction of hydrophobic steroids with dehydrated, hydrophobized PIPAAm-grafted capillary surfaces was observed above 30 degrees C without any column packing materials. Steroid elution behavior from PIPAAm-grafted capillaries contrasted sharply with that from PIPAAm hydrogel grafted porous monolithic silica capillaries prepared by electron beam (EB) irradiation wherein significant peak broadening was observed at high-temperature regardless of sample hydrophobicity factors (log P values), indicating multistep separation modes in coated monolithic silica capillaries. In conclusion, thermoresponsive polymer-grafted capillary inner wall surfaces prepared by ATRP exhibit useful temperature-dependent surface property alterations effective to regulate interactions with biomolecules without requirements for separation bed packing materials within the capillary lumen. PMID- 16378456 TI - QCM studies of gel spreading: Kraton gels on polystyrene surfaces. AB - Contact of a polymer gel made from a styrene/ethylene-butene/styrene triblock copolymer in mineral oil was investigated by bringing the gel into contact with the coated surface of a quartz crystal microbalance (QCM). The experimental apparatus enabled simultaneous measurement of the load, displacement, and contact area, in addition to the resonant frequency and dissipation of the oscillating quartz crystal. The QCM response was determined by the linear viscoelastic properties of the gel at the frequency of oscillation. A geometric correction factor involving the contact area provided a means for quantitatively determining these viscoelastic parameters as the gel spread over the QCM surface. When the gel was removed from the surface, a thin solvent layer was left behind. The thickness of this solvent layer was determined from the QCM response and was compared to predictions from a simple model involving the disjoining pressure of the film and the osmotic pressure of the gel. Qualitative agreement with the model required that tensile, adhesive forces at the perimeter of the gel/QCM contact area were taken into account when calculating the film thickness. PMID- 16378457 TI - Phase domain of the cubic Im3m mesoporous silica in the EO106PO70EO106-butanol H2O system. AB - n-butanol has been chosen as an organic additive in the SiO(2) EO(106)PO(70)EO(106)-H(2)O system at low HCl concentrations, for the generation of large-pore mesoporous silicas with easily tailored textural and structural properties. As opposed to previous reports on syntheses of cubic mesoporous silica that are usually performed in a very narrow composition range, we report now the possibility of preparing large-pore cagelike mesoporous silicas in a wide range of synthesis mixture compositions. Particularly, the cubic Imm silica with large interconnected cagelike pores (SBA-16) can easily be synthesized with controlled pore sizes and wall thicknesses, depending upon the synthesis mixture composition. The primary mesopore volume of the SBA-16 cages can be tuned from 0.27 to 0.56 cm(3) g(-)(1), and the mesopore size is shown to range from 4.7 to 7.2 nm, by performing a simple adjustment of the starting mixture composition. With the synthesis parameters varied, we describe the first complete diagram of the product phase domains obtained for silica mesophases in a SiO(2) EO(106)PO(70)EO(106)-butanol-H(2)O system. Other ordered mesophases also observed in this system are the face-centered cubic Fmm silica mesophase and a 2D hexagonal-like mesostructure. Importantly here, the use of a low acid catalyst concentration regime allowed the preparation of silica mesophases in almost thermodynamically controlled conditions because of slow condensation kinetics of the inorganics. Such conditions enabled the introduction of n-butanol as the phase-controlling agent in the system, providing efficient tuning of the mesophase topology. The description of the phase domains provides a future basis for the design of large-pore mesoporous silicas with tailored textural and structural properties. Mesoporous samples obtained within the composition ranges of the phase domains are characterized by powder X-ray diffraction (PXRD) and nitrogen physisorption measurements. PMID- 16378458 TI - Impact of steric interactions on the helical transition in assemblies of discotic molecules. AB - We investigate theoretically the effect of excluded-volume interactions on the helical configurational transition of supramolecular assemblies in solutions of chiral disklike molecules. To this end, we set up a second-virial theory within the context of the helical self-assembly of rodlike objects. We find that interaggregate interactions shift both the helical-transition point and the sharpness of the transition. For realistic values of the model parameters, the helical-transition temperature shifts by several degrees, and the more so the higher the concentration of assembling material. The mean aggregation number is also affected by the interactions, albeit only by a modest amount, unless the solution becomes very concentrated. PMID- 16378459 TI - X-ray and neutron reflectometry study of glow-discharge plasma polymer films. AB - Radio-frequency glow-discharge plasma polymer thin films of allylamine (AA) and hexamethyldisiloxane (HMDSO) were prepared on silicon wafers and analyzed by a combination of X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), X-ray reflectometry (XRR), and neutron reflectometry (NR). AFM and XRR measurements revealed uniform, smooth, defect-free films of 20-30 nm thickness. XPS measurements gave compositional data on all elements in the films with the exception of hydrogen. In combination with XRR and NR, the film composition and mass densities (1.46 and 1.09 g cm(-)(3) for AA and HMDSO, respectively) were estimated. Further NR measurements were conducted with the AA and HMDSO films in contact with water at neutral pH. Three different H(2)O/D(2)O mixtures were used to vary the contrast between the aqueous phase and the polymer. The amount of water penetrating the film, as well as the number of labile protons present, was determined. The AA film in contact with water was found to swell by approximately 5%, contain approximately 3% water, and have approximately 24% labile protons. The HDMSO polymer was found to have approximately 6% labile protons, no thickness increase when in contact with water, and essentially no solvent penetration into the film. The difference in the degree of proton exchange within the films was attributed to the substantially different surface and bulk chemistries of the two films. PMID- 16378460 TI - Influence of shell thickness and cross-link density on the structure of temperature-sensitive poly-N-isopropylacrylamide-poly-N-isopropylmethacrylamide core-shell microgels investigated by small-angle neutron scattering. AB - Swelling properties of doubly temperature sensitive core-shell microgels consisting of two thermosensitive polymers with lower critical solution temperatures (LCTS) at, respectively, 34 degrees C in the core and 44 degrees C in the shell have been investigated by small-angle neutron scattering (SANS). A core-shell form factor has been employed to evaluate the structure, and the real space particle structure is expressed by radial density profiles. By this means, the influences of both shell/core mass composition and shell cross-linker content on the internal structure have been revealed at temperatures above, between, and below the LCSTs. Higher shell/core mass ratios lead to an increased expansion of the core at temperatures between the LCSTs, whereas a variation of cross-linker in the shell mainly effects the dimensions of the shell. The influence on the core structure was interpreted as resulting from an elastic force developed from the swollen shell. At temperatures below the core LCST, the core cannot swell to its native size (i.e., in the absence of a shell), because the maximum expanded shell network prohibits further swelling. Thus, depending on temperature, the shell either expands or compresses the core. PMID- 16378461 TI - Ion-exchange properties and electrochemical characterization of quaternary ammonium-functionalized silica microspheres obtained by the surfactant template route. AB - Porous silica spheres functionalized with quaternary ammonium groups have been prepared by co-condensation of N-((trimethoxysilyl)propyl)-N,N,N trimethylammonium chloride (TMTMAC) and tetraethoxysilane (TEOS) in the presence of cetyltrimethylammonium as a template and ammonia as a catalyst. The physicochemical characteristics of the resulting ion exchangers have been analyzed by various techniques and discussed with respect to the amount of organofunctional groups in the materials. For comparison purposes, both an ordered MCM-41 type mesoporous silica and two silica gels of different pore size have been grafted with TMTMAC. The ion-exchange capabilities were first evaluated from batch experiments (determination of anion-exchange capacities) and then by ion-exchange voltammetry at carbon paste electrodes modified with these hybrid materials. Effective concentration of Fe(CN)(6)(3)(-) species in the anion exchangers was pointed out, while no significant accumulation of Ru(NH(3))(6)(3+) was observed. The preconcentration efficiency was discussed on the basis of the organic group content in the materials as well as their structure and porosity. A second series of materials displaying zwitterionic surfaces was finally prepared and characterized with respect to their physicochemical properties and ion exchange voltammetric behavior. They consisted of sulfonic acid-functionalized mesoporous silica samples resulting from the oxidation of thiol-functionalized silica spheres obtained by co-condensation of mercaptopropyl-trimethoxysilane (MPTMS) and TEOS, which were then grafted with TMTMAC at various functionalization levels. Possible interactions between the ammonium and sulfonate moieties in the confined medium were pointed out from X-ray photoelectron spectroscopy. The competitive accumulation-rejection of Fe(CN)(6)(3)(-) and Ru(NH(3))(6)(3+) redox probes was finally studied by cyclic voltammetry. PMID- 16378462 TI - Electrodes modified with the phase transition polymer and heme peptide: biocatalysis and biosensing with tunable activity and dynamic range. AB - An electrode was modified with a phase transition polymer, poly(N isopropylacrylamide), and the polymer was further modified with a peroxidase model compound, heme peptide (HP). As the polymer layer shrank at temperatures above 30-40 degrees C, the catalytic activity of the HP molecules for H(2)O(2) reduction improved, and simultaneously, the number of HP molecules that can communicate electrochemically with the electrode increased. As a result, the catalytic current for H(2)O(2) reduction in the shrunken state was 4 times larger than that in the swollen state. This reversible change was exploited for tuning the sensitivity and dynamic range of the HP electrode in H(2)O(2) biosensing. The dynamic range in inhibition-based biosensing of imidazole derivatives was also tunable. PMID- 16378463 TI - Model description of contact angles in electrowetting on dielectric layers. AB - The electrowetting on dielectric (EWOD) technique has considerable potential for microfluidic and biomedical applications. The Lippmann-Young model based on the force balance concept has long been used to predict the contact angles of droplets under electrowetting. However, recent experimental evidence has indicated that this model fails to provide accurate predictions of the lower contact angles associated with saturation conditions at higher electric potentials. Hence, the study simulates the internal flow in an actuated droplet and treats it as stagnation-point flow. This kinetic energy is then taken into consideration while calculating the contact angles using an energy balance model. The energy of an actuated droplet is contributed by the combination of the side surface tension energy, the base tension energy, the dielectric energy, and the kinetic energy when deriving the energy balance model. Consequently, the new energy balance model modifies the Lippmann-Young equation, thereby providing enhanced reasonable predictions of the droplet contact angle across the higher electric potential where the contact angles are close to the saturated condition. PMID- 16378464 TI - Rhodium complex with ethylene ligands supported on highly dehydroxylated MgO: synthesis, characterization, and reactivity. AB - Mononuclear rhodium complexes with reactive olefin ligands, supported on MgO powder, were synthesized by chemisorption of Rh(C(2)H(4))(2)(C(5)H(7)O(2)) and characterized by infrared (IR), (13)C MAS NMR, and extended X-ray absorption fine structure (EXAFS) spectroscopies. IR spectra show that the precursor adsorbed on MgO with dissociation of acetylacetonate ligand from rhodium, with the ethylene ligands remaining bound to the rhodium, as confirmed by the NMR spectra. EXAFS spectra give no evidence of Rh-Rh contributions, indicating that site-isolated mononuclear rhodium species formed on the support. The EXAFS data also show that the mononuclear complex was bonded to the support by two Rh-O bonds, at a distance of 2.18 A, which is typical of group 8 metals bonded to oxide supports. This is the first simple and nearly uniform supported mononuclear rhodium-olefin complex, and it appears to be a close analogue of molecular catalysts for olefin hydrogenation in solution. Correspondingly, the ethylene ligands bonded to rhodium in the supported complex were observed to react with H(2) to form ethane, and the supported complex was catalytically active for the ethylene hydrogenation at 298 K. The ethylene ligands also underwent facile exchange with C(2)D(4), and exposure of the sample to carbon monoxide led to the formation of rhodium gem dicarbonyls. PMID- 16378465 TI - Tailoring surface properties to build colloidal diagnostic devices: controlling interparticle associations. AB - The ability to control the surface properties and subsequent colloidal stability of dispersed particles has widespread applicability in many fields. Sub micrometer fluorescent silica particles (reporters) can be used to actively encode the combinatorial synthesis of peptide libraries through interparticle association. To achieve these associations, the surface chemistry of the small fluorescent silica reporters is tailored to encourage robust adhesion to large silica microparticles onto which the peptides are synthesized. The interparticle association must withstand a harsh solvent environment, multiple synthetic and washing procedures, and biological screening buffers. The encoded support beads were exposed to different solvents used for peptide synthesis, and different solutions used for biological screening including phosphate buffered saline (PBS), 2-[N-morpholino]ethane sulfonic acid (MES) and a mixture of MES and N-(3 dimethyl-aminopropyl)-N'-ethylcarbodiimide (EDC). The number of reporters remaining adhered to the support bead was quantified after each step. The nature of the associations were explored and tested to optimize the efficiency of these phenomena. Results presented illustrate the influence of the surface functionality and polyelectrolyte modification of the reporters. These parameters were investigated through zeta potential and X-ray photoelectron spectroscopy. PMID- 16378466 TI - Electrochemical microbead-based immunoassay using an (eta5 cyclopentadienyl)tricarbonylmanganese redox marker bound to bovine serum albumin. AB - A first example of the solid-phase immunoassay of a high-weight antigen bovine serum albumin (BSA) using an (eta(5)-cyclopentadienyl)tricarbonylmanganese (cymantrene) redox probe is presented. The electrochemical detection is based on the impedance measurements of a one-electron reversible reduction of the organometallic probe. The microbead-based immunoassay is discussed for two types of microbeads with different diameters (2.5 and 90 microm) and capabilities to bind the immunoglobulins (2.4 and 10 microg/mg of beads). The use of larger agarose microbeads allows the formation of an antigen-antibody complex at the surface of microbeads directly dispersed in the analyzed solution. No additional separation step is necessary for the electrochemical competitive immunoassay analysis of BSA. The presence of agarose beads in the analyzed solution has no effect on the electrochemical signal from labeled BSA released from the antigen antibody complex. PMID- 16378470 TI - Speech motor development during acquisition of the voicing contrast. AB - Lip and jaw movements were studied longitudinally in 19-month-old children as they acquired the voicing contrast for /p/ and /b/. A movement tracking system obtained lip and jaw kinematics as participants produced the target utterances /papa/ and /baba/. Laryngeal adjustments were also tracked through acoustically recorded voice onset time (VOT) of the consonants. Across this period of developmental phonological change, the children began to produce VOTs in 2 distinct categories for voiced and voiceless plosives. Specific kinematic differences were observed during oral opening and closing and between spatial and temporal parameters of movement. The development of the voicing contrast was most closely associated with changes in jaw kinematics for oral opening in comparison to that of the lip. Conversely, movements into oral closing were not accompanied by significant increases in jaw, upper lip, or lower lip displacement or velocity, although a decrease in jaw movement variability was found. There was no evidence of phoneme-specific movement differences between /p/ and /b/ in the children or in the adults studied. Spatial coupling between the jaw and upper lip changed significantly across sessions, whereas changes in temporal coupling were not observed. Findings indicate that oral opening and closing have different task requirements and that children modify their articulatory movements to meet the demands of each task. Overall, the findings illustrate how orofacial movements and laryngeal function change in parallel during linguistic development. PMID- 16378471 TI - Sex-related acoustic changes in voiceless English fricatives. AB - This investigation is a comprehensive acoustic study of 4 voiceless fricatives (/f theta s /) in English produced by adults and pre- and postpubescent children aged 6-14 years. Vowel duration, amplitude, and several different spectral measures (including spectral tilt and spectral moments) were examined. Of specific interest was the pattern of normal development of the acoustic properties of fricatives and the nature of sex-specific patterns of fricative articulation in prepubescent children. Little evidence of amplitude or duration differences was found between speakers that was related to sex of the speaker. However, significant sex-specific differences in fricative articulation were found in all groups of speakers-even in the youngest children (ages 6-7 years) although there was an indication that some of the acoustic differences between females and males is reduced or absent in the youngest children. Results from discriminant analysis demonstrated that a discriminant function based on the adult male tokens was generally better at classifying fricatives produced by male speakers than female speakers, regardless of age. This showed that sex-related differences (presumably a function of sex-linked vocal tract variation) were present even in the youngest speaker group. However, the classification accuracy of the female model showed a steady improvement with the increased age of the female speakers and may provide support for the claim that sex-related developmental differences may just be emerging in the youngest age group. PMID- 16378472 TI - Impact of aberrant acoustic properties on the perception of sound quality in electrolarynx speech. AB - A large percentage of patients who have undergone laryngectomy to treat advanced laryngeal cancer rely on an electrolarynx (EL) to communicate verbally. Although serviceable, EL speech is plagued by shortcomings in both sound quality and intelligibility. This study sought to better quantify the relative contributions of previously identified acoustic abnormalities to the perception of degraded quality in EL speech. Ten normal listeners evaluated the sound quality of EL speech tokens that had been acoustically enhanced by (a) increased low-frequency energy, (b) EL-noise reduction, and (c) fundamental frequency variation to mimic normal pitch intonation in relation to nonenhanced EL speech, normal speech, and normal monotonous speech (fundamental frequency variation removed). In comparing all possible combinations of token pairs, listeners were asked to identify which one of each pair sounded most like normal natural speech, and then to rate on a visual analog scale how different the chosen token was from normal speech. The results indicate that although EL speech can be most improved by removing the EL noise and providing proper pitch information, the resulting quality is still well below that of normal natural speech or even that of monotonous natural speech. This suggests that, in addition to the widely acknowledged acoustic abnormalities examined in this investigation, there are other attributes that contribute significantly to the unnatural quality of EL speech. Such additional factors need to be clearly identified and remedied before EL speech can be made to more closely approximate the sound quality of normal natural speech. PMID- 16378473 TI - Adaptation of a Pocket PC for use as a wearable voice dosimeter. AB - This article deals with the adaptation of a commercially available Pocket PC for use as a voice dosimeter, a wearable device that measures the vocal dose of teachers or other individuals on the job, at home, and elsewhere during the course of an entire day. An engineering approach for designing a voice dosimeter is described, and design data are presented. Technical issues include transducer selection, dynamic range, frequency response, memory requirements, power requirements, attachment, cables, connections, and data collection. Advantages and disadvantages of the design are discussed. PMID- 16378474 TI - Low expressive vocabulary: higher heritability as a function of more severe cases. AB - This study of 4,274 pairs of 4-year-old twins from the Twins Early Development Study explored the magnitude of genetic and environmental effects on low expressive vocabulary skill, both as a function of general cognitive ability and as a function of the severity of expressive vocabulary impairment. Assessments were conducted through parent report measures. Two types of vocabulary deficit were identified: low vocabulary paired with typical general cognition (i.e., specific expressive vocabulary impairment) and low vocabulary paired with low general cognition (i.e., nonspecific expressive vocabulary impairment). The magnitude of genetic and environmental effects on low expressive vocabulary skill did not differ for these 2 types of expressive vocabulary deficit. By systematically varying the cutoffs used to define vocabulary and general cognitive delay, potential changes in the magnitude of genetic and environmental effects were examined. Results suggested that the severity of vocabulary deficit rather than level of cognitive functioning was a more meaningful etiological distinction: The heritability of low expressive vocabulary was higher and the influence of shared environment lower, as increasingly severe vocabulary deficits were identified. Implications for molecular genetics and the construct of specific language deficits are discussed. PMID- 16378475 TI - An investigation of speech perception in children with specific language impairment on a continuum of formant transition duration. AB - Fifteen children diagnosed with specific language impairment (SLI) and 15 typically developing (TD) children were tested for identification performance on 2 synthetic speech continua varying in formant transition durations (FTDs). One continuum varied from /ba/ to /wa/, and the other varied from /da/ to /ja/. Various d'-related measures from signal detection theory were used to compare category boundaries and indirectly derive sensitivity to phonetic changes in category tokens along each continuum. The SLI group showed less consistent identification performance along the /ba/-/wa/ series relative to the TD group, as well as reduced sensitivity to phonetic changes along the continuum. On the /da/-/ja/ series, the SLI group revealed less consistent identification performance on the short FTD end but similar identification levels to the TD group at the long FTD end. The overall results support the contention that children with SLI reveal a deficiency in the processing of speech sounds at the level of segmental identity. PMID- 16378476 TI - A phonemic implicational feature hierarchy of phonological contrasts for English speaking children. AB - Contrastive feature hierarchies have been developed and used for some time in depicting typical phonological development and in guiding therapy decisions. Previous descriptions of feature use have been based on independent analyses and usually phonetic inventories. However, recent trends in phonology include a relational analysis of phonemic inventories (D. Ingram & K. D. Ingram, 2001). The current investigation was a relational analysis of the phonemic inventories of 40 typically developing 2-year-old American-English-speaking children. Consonant inventories were derived from spontaneous speech samples using the Logical International Phonetics Programs computer software (D. K. Oller & R. E. Delgado, 1999). Cluster analysis was used to determine the grouping of contrastive features. Four levels emerged. Level I included [consonant], [sonorant], and [coronal], Level II included [voice], Level III included [anterior], [continuant], and [nasal], and Level IV included [lateral] and [strident]. Results suggested that the resulting 4-level phonemic feature hierarchy might be used to classify the phonological systems of children with phonological disorders. PMID- 16378477 TI - Toward diagnostic and phenotype markers for genetically transmitted speech delay. AB - Converging evidence supports the hypothesis that the most common subtype of childhood speech sound disorder (SSD) of currently unknown origin is genetically transmitted. We report the first findings toward a set of diagnostic markers to differentiate this proposed etiological subtype (provisionally termed speech delay-genetic) from other proposed subtypes of SSD of unknown origin. Conversational speech samples from 72 preschool children with speech delay of unknown origin from 3 research centers were selected from an audio archive. Participants differed on the number of biological, nuclear family members (0 or 2+) classified as positive for current and/or prior speech-language disorder. Although participants in the 2 groups were found to have similar speech competence, as indexed by their Percentage of Consonants Correct scores, their speech error patterns differed significantly in 3 ways. Compared with children who may have reduced genetic load for speech delay (no affected nuclear family members), children with possibly higher genetic load (2+ affected members) had (a) a significantly higher proportion of relative omission errors on the Late-8 consonants; (b) a significantly lower proportion of relative distortion errors on these consonants, particularly on the sibilant fricatives /s/, /z/, and //; and (c) a significantly lower proportion of backed /s/ distortions, as assessed by both perceptual and acoustic methods. Machine learning routines identified a 3 part classification rule that included differential weightings of these variables. The classification rule had diagnostic accuracy value of 0.83 (95% confidence limits = 0.74-0.92), with positive and negative likelihood ratios of 9.6 (95% confidence limits = 3.1-29.9) and 0.40 (95% confidence limits = 0.24 0.68), respectively. The diagnostic accuracy findings are viewed as promising. The error pattern for this proposed subtype of SSD is viewed as consistent with the cognitive-linguistic processing deficits that have been reported for genetically transmitted verbal disorders. PMID- 16378478 TI - The effect of age at cochlear implant initial stimulation on expressive language growth in infants and toddlers. AB - This study examined the growth of expressive language skills in children who received cochlear implants (CIs) in infancy. Repeated language measures were gathered from 29 children who received CIs between 10 and 40 months of age. Both cross-sectional and growth curve analyses were used to assess the relationship between expressive language outcomes and CI experience. A beneficial effect of earlier implantation on expressive language growth was found. Growth curve analysis showed that growth was more rapid in children implanted as infants than those implanted as toddlers. Age at initial stimulation accounted for 14.6% of the variance of the individual differences in expressive language growth rates. PMID- 16378479 TI - Roles of initiation and responsiveness in access and participation for children with specific language impairment. AB - This study compared the abilities of children with specific language impairment (SLI; n = 10) and typically developing (TD) children (n = 13) to access and participate in an ongoing interaction between two unfamiliar peer partners. Results revealed that all children in the study accessed by either making an unprompted initiation toward their peers (access initiation) or by responding to a question or play invitation directed toward them (access response). However, 4 children with SLI were unsuccessful in achieving successful access initiation during the 10-min play period. Children with SLI required a longer period of time to achieve access initiation. Following access, children with SLI were addressed significantly less by their play partners, participated in less group play, and engaged in more individual play and onlooking behavior. Among the SLI group, language levels were negatively related to the time children required to achieve their first successful access and first access initiation. Expressive language levels were positively related to the percentage of utterances children produced postaccess and the percentage of utterances they were addressed postaccess by their play partners. Differences in receptive skills among SLI children were less strongly related to the time they required in achieving their first access and were unrelated to their ability to participate in the interaction. PMID- 16378480 TI - Exploring the processing continuum of single-word comprehension in aphasia. AB - This study investigated the vulnerability of lexical processing in individuals with aphasia. Though classical teaching of aphasia syndromes holds that people with Broca's aphasia have intact comprehension at the single-word level, the nature and extent of this purported sparing were explored under suboptimal processing conditions. A combination of acoustic distortions (low-pass filtering and time compression) was used to probe for "break points" in lexical comprehension in a group of individuals with aphasia. Results suggest that accurate and efficient lexical processing is vulnerable to suboptimal listening climates, and that processing under these conditions reveals the continuous nature of the impairment of linguistic behaviors observed in individuals with aphasia. PMID- 16378481 TI - Beyond capacity limitations: determinants of word recall performance on verbal working memory span tasks in children with SLI. AB - Reduced verbal working memory capacity has been proposed as a possible account of language impairments in specific language impairment (SLI). Studies have shown, however, that differences in strength of linguistic representations in the form of word frequency affect list recall and performance on verbal working memory tasks. This suggests that verbal memory capacity and long-term linguistic knowledge may not be distinct constructs. It has been suggested that linguistic representations in SLI are weak in ways that result in a breakdown in language processing on tasks that require manipulation of unfamiliar material. In this study, the effects of word frequency, long-term linguistic knowledge, and serial order position on recall performance in the competing language processing task (CLPT) were investigated in 10 children with SLI and 10 age-matched peers (age 8 years 6 months to 12 years 4 months). The children with SLI recalled significantly fewer target words on the CLPT as compared with their age-matched controls. The SLI group did not differ, however, in their ability to recall target words having high word frequency but were significantly poorer in their ability to recall words on the CLPT having low word frequency. Differences in receptive and expressive language abilities also appeared closely related to performance on the CLPT, suggesting that working memory capacity is not distinct from language knowledge and that degraded linguistic representations may have an effect on performance on verbal working memory span tasks in children with SLI. PMID- 16378482 TI - Perception of spectral contrast by hearing-impaired listeners. AB - The ability to discriminate the spectral shapes of complex sounds is critical to accurate speech perception. Part of the difficulty experienced by listeners with hearing loss in understanding speech sounds in noise may be related to a smearing of the internal representation of the spectral peaks and valleys because of the loss of sensitivity and an accompanying reduction in frequency resolution. This study examined the discrimination by hearing-impaired listeners of highly similar harmonic complexes with a single spectral peak located in 1 of 3 frequency regions. The minimum level difference between peak and background harmonics required to discriminate a small change in the spectral center of the peak was measured for peaks located near 2, 3, or 4 kHz. Component phases were selected according to an algorithm thought to produce either highly modulated (positive Schroeder) or very flat (negative Schroeder) internal waveform envelopes in the cochlea. The mean amplitude difference between a spectral peak and the background components required for discrimination of pairs of harmonic complexes (spectral contrast threshold) was from 4 to 19 dB greater for listeners with hearing impairment than for a control group of listeners with normal hearing. In normal hearing listeners, improvements in threshold were seen with increasing stimulus level, and there was a strong effect of stimulus phase, as the positive Schroeder stimuli always produced lower thresholds than the negative Schroeder stimuli. The listeners with hearing loss showed no consistent spectral contrast effects due to stimulus phase and also showed little improvement with increasing stimulus level, once their sensitivity loss was overcome. The lack of phase and level effects may be a result of the more linear processing occurring in impaired ears, producing poorer-than-normal frequency resolution, a loss of gain for low amplitudes, and an altered cochlear phase characteristic in regions of damage. PMID- 16378483 TI - Multiple looks in speech sound discrimination in adults. AB - N. F. Viemeister and G. H. Wakefield's (1991) multiple looks hypothesis is a theoretical approach from the psychoacoustic literature that has promise for bridging the gap between results from speech perception research and results from psychoacoustic research. This hypothesis accounts for sensory detection data and predicts that if the "looks" at a stimulus are independent and information is combined optimally, sensitivity should increase for 2 pulses relative to 1 pulse. Specifically, d' (a bias-free measure of sensitivity) for 2 pulses should be larger than d' for 1 pulse. One speech discrimination paradigm that presents stimuli with multiple presentations is the change/no-change procedure. On a change trial, the standard and comparison stimuli differ; on a no-change trial, they are the same. Normal-hearing adults were tested using the change/no-change procedure with 3 consonant-vowel minimal pairs in combinations of 1, 2, and 4 repetitions of standard and comparison stimuli at various signal-to-noise ratios. If multiple looks extend to this procedure, performance should increase with higher repetition numbers. Performance increased with more presentations of the speech contrasts tested. The multiple looks hypothesis predicted performance better at low repetition numbers when performance was near d' values of 1.0 than at higher repetition numbers and higher performance levels. PMID- 16378484 TI - Categorical perception of speech by children with specific language impairments. AB - Previous research has suggested that children with specific language impairments (SLI) have deficits in basic speech perception abilities, and this may be an underlying source of their linguistic deficits. These findings have come from studies in which perception of synthetic versions of meaningless syllables was typically examined in tasks with high memory demands. In this study, 20 children with SLI (mean age = 9 years, 3 months) and 20 age-matched peers participated in a categorical perception task. Children identified and discriminated digitally edited versions of naturally spoken real words in tasks designed to minimize memory requirements. Both groups exhibited all hallmarks of categorical perception: a sharp labeling function, discontinuous discrimination performance, and discrimination predicted from identification. There were no group differences for identification data, but children with SLI showed lower peak discrimination values. Children with SLI still discriminated phonemically contrastive pairs at levels significantly better than chance, with discrimination of same-label pairs at chance. These data suggest that children with SLI perceive natural speech tokens comparably to age-matched controls when listening to words under conditions that minimize memory load. Further, poor performance on speech perception tasks may not be due to a speech perception deficit, but rather to a consequence of task demands. PMID- 16378485 TI - Developmental role of static, dynamic, and contextual cues in speech perception. AB - The purpose of the current study was to examine the role of syllable duration context as well as static and dynamic acoustic properties in child and adult speech perception. Ten adults and eleven 4-5-year-old children identified a syllable as [ba] or [wa] (stop-glide contrast) in 3 conditions differing in synthetic continua. The 1st condition tested the potential existence of the syllable duration effect in young children, whereas the 2nd and 3rd conditions examined the developmental role of static and dynamic cues, respectively, as related to syllable duration context effects. In the 1st condition, the 1st and 2nd formant transition duration of stimuli varied from those appropriate for [ba] to those appropriate for [wa]. For the 2nd condition, a static burst was added to Condition 1 stimuli. For the 3rd condition, the dynamic transition frequency and transition duration for the first 3 formants varied as appropriate for [ba] and [wa]. In each condition, 3 syllable context durations of 105 ms, 170 ms, and 315 ms were tested. The results indicated that syllable duration context effects were present across all conditions for both adults and children. However, the adults and children did differ in the 3rd condition, in which both the transition frequency and the transition duration were altered. Thus, children used the dynamic formant transitions differently than adults when transition frequency was varied along with transition duration. These findings show that children have a bias toward formant transitions and indicate that young children of 4-5 years of age attend differently than adults to changes in dynamic cues such as formant transitions, as predicted by the developmental cue weighting shift model. PMID- 16378487 TI - Metathesis reactions. General considerations. AB - General considerations on the metathesis reaction are reported and illustrated by examples in the area of natural products and/or biologically active compounds. PMID- 16378488 TI - Recent applications of olefin ring-closing metathesis (RCM) in the synthesis of biologically important alkaloids, terpenoids, polyketides and other secondary metabolites. AB - With the commercial availability of well-defined ruthenium metathesis catalysts which combine high stability and broad functional group compatibility with undiminished activity, ring-closing metathesis (RCM) is now routinely integrated in the planning of natural product syntheses. Thus, ring sizes of practically any kind from five onward may be formed. The presence of heteroatoms such as oxygen and nitrogen is of minor importance. Ring-strain and the presence of additional double bonds may be overcome by judicious selection of conditions. A major drawback of macrocyclic RCM still is the lack of E/Z stereocontrol. Nevertheless, the current overview will demonstrate the overwhelming power and scope of RCM in the total synthesis of structurally demanding natural products. PMID- 16378489 TI - The application of olefin metathesis to the synthesis of biologically active macrocyclic agents. AB - Recent developments of the metathesis reaction in the area of biologically active molecules are presented. Scope and limitations of ring-closing metathesis to form medium and large rings are discussed and illustrated by the epothilone synthesis. Applications of the metathesis reaction related to medicinal chemistry, including solid phase synthesis and combinatorial chemistry are presented. PMID- 16378490 TI - Ring closing metathesis in the synthesis of biologically interesting peptidomimetics, sugars and alkaloids. AB - Olefin metathesis has rapidly established itself as an essential tool in the synthetic chemist's armoury. The ease of operation and functional group tolerance that is obtained with the modern generation of catalysts makes the use of metathesis an extremely attractive option when preparing medicinally interesting molecules. This article will outline some of the ways in which chemists from both industry and academia have been utilising and developing metathesis in the search for novel biological probes and drug leads. PMID- 16378491 TI - Olefin metathesis route to antiviral nucleosides. AB - The success of the early nucleoside agents, the toxicity and metabolic instability of many nucleoside analogues and the effects of viral pathogens on public health are driving the design, synthesis and evaluation of new nucleoside analogues. In this context, a powerful reaction has emerged over the past decade that has fundamentally changed the outlook on nucleoside chemistry: the olefin metathesis reaction. This review is designed to give an overview of the synthesis of some nucleosides of biological interest, according to their structural types (e.g., neplanocins and aristeromycin analogues, 2',3'-unsaturated nucleoside analogues, and acyclonucleosides), using metathesis reactions by employing either the alkoxy imido molybdenum catalyst developed by Schrock and various ruthenium carbene catalysts developed by Grubbs, Hoveyda-Grubbs and Nolan. PMID- 16378492 TI - Application of olefin cross-metathesis to the synthesis of biologically active natural products. AB - An overview of the use of olefin cross-metathesis in the synthesis of biologically active natural products is presented. The diverse examples are organized according to the outcome of the olefin constructed by the cross metathesis reaction: this olefin can be either present in the final product, reduced, engaged in other transformations, or involved in tandem processes. PMID- 16378493 TI - Cross-metathesis: efficient preparation of trialkyl-substituted isoprenoid olefins as key-intermediates for tocopherol synthesis. AB - A summary of the application of ruthenium catalyzed olefin cross-metathesis towards the synthesis of tocopherols (vitamin E) is given. This group of biologically important fat-soluble antioxidants is synthetically available by various routes, for which key-intermediates containing trialkyl-substituted olefinic double bonds can now be prepared efficiently. The results presented may be of interest for the area of syntheses of isoprenoid natural products in general. PMID- 16378495 TI - Therapeutic targets in inflammatory disease. AB - This commentary is an introduction to a special issue on "Latest Developments in the Treatment of Inflammation". It outlines some key events in the inflammatory response to infection or injury and describes some of the important drug targets of relevance to the succeeding articles, which survey inhibitors of these targets as prospective or current antiinflammatory drugs. It also highlights important limitations in the validation of inflammatory drug targets, and in the rate of discovery and development of new antiinflammatory drugs. PMID- 16378496 TI - Drugs to treat inflammation: a historical introduction. AB - Drugs to treat inflammation are discussed under the following headings: (1) random discoveries covering copper, salicylates, heterocyclic diones, ACTH, adrenal steroids and disease-modifying agents (DMARDs); these include Au(I) thiolates, chloroquine, and hydroxychloroquine, minocycline, cyclosporin, salazopyrine, D-penicillamine and methotrexate; (2) programmed NSAID developments covering salicylates and fenamates, arylalkanoates, diones, non-acidic NSAIDs, clozic, lobenzarit and coxibs; (3) synthetic glucocorticosteroids; and (4) 'Biologicals' for neutralising pro-inflammatory cytokines. Clinical problems are highlighted, particularly unacceptable side-effects affecting the GI tract, skin, liver, etc. that caused many drugs to be withdrawn. Drug combinations may overcome some of these problems. The bibliography has selected reviews and monographs covering 50 years of publications. PMID- 16378497 TI - Antibody treatments of inflammatory arthritis. AB - Inflammatory arthropathies such as rheumatoid arthritis, ankylosing spondylitis, and psoriatic arthritis are extremely common in the community, with a prevalence of up to 5%, and they cause substantial morbidity. The development of anti-TNF agents for use initially in rheumatoid arthritis, and subsequently more broadly in inflammatory arthritis, represents the biggest advance in management of these conditions since the introduction of corticosteroid agents, and is a major vindication of public funded arthritis research. However, there are limitations of even these highly effective agents. A significant minority of patients with inflammatory arthritis do not respond to these anti-TNF agents, they are associated with substantial risk of toxicity, require parenteral administration, and are extremely expensive. New antibody treatments in development can be divided into anti-cytokine agents, cell-targeted therapies, co-stimulation inhibitors, and treatments aimed at preventing joint erosion consequent on inflammation. This review discusses the state of the art in the development of these agents for management of this common group of diseases. PMID- 16378498 TI - Anti-inflammatory immunotherapy for multiple sclerosis/experimental autoimmune encephalomyelitis (EAE) disease. AB - Multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), are inflammatory diseases of the central nervous system (CNS) characterized by localized areas with demyelination. Disease is believed to be an autoimmune disorder mediated by activated immune cells such as T- and B lymphocytes and macrophages/microglia. Lymphocytes are primed in the peripheral tissues by antigens, and clonally expanded cells infiltrate the CNS. They produce large amounts of inflammatory cytokines, nitric oxide (NO) that lead to demyelination and axonal degeneration. Although several studies have shown that oligodendrocytes (OLGs), the myelin-forming glial cells in the CNS, are sensitive to cell death stimuli, such as cytotoxic cytokines, anti-myelin antibodies, NO, and oxidative stress, in vitro, the mechanisms underlying injury to the OLGs in MS/EAE remain unclear. The central role of glutamate receptors in mediating excitotoxic neuronal death in stroke, epilepsy, trauma and MS has been well established. Glutamate is the major excitatory amino acid transmitter within the CNS and it's signaling is mediated by a number of postsynaptic ionotropic and metabotropic receptors. Inflammation can be blocked with anti-cell adhesion molecules MAb, simultaneously protected oligodendrocytes and neurons against glutamate-mediated damage with the AMPA/kainate antagonist NBQX, and the NMDA receptor antagonist GPE, could thus be effective therapies for multiple sclerosis. PMID- 16378499 TI - Inhibitors of TACE and Caspase-1 as anti-inflammatory drugs. AB - TNF-alpha neutralising agents such as Infliximab (Remicade), Etanercept (Enbrel) and the IL-1 receptor antagonist Anakinra (Kineret), are currently used clinically for the treatment of many inflammatory diseases such as Crohn's disease, rheumatoid arthritis, ankylosing spondylitis, juvenile rheumatoid arthritis, psoriatic arthritis and psoriasis. These protein preparations are expensive to manufacture and administer, need to be injected and can cause allergic reactions. An alternative approach to lowering the levels of TNF-alpha and IL-1beta in inflammatory disease, is to inhibit the enzymes that generate these cytokines using cheaper small molecules. This paper is a broad overview of the progress that has been achieved so far, with respect to small molecule inhibitor design and pharmacological studies (in animals and humans), for the metalloprotease Tumour Necrosis Factor-alpha Converting Enzyme (TACE) and the cysteine protease Caspase-1 (Interleukin-1beta Converting Enzyme, ICE). Inhibitors of these two enzymes are currently considered to be good therapeutic targets that have the potential to provide relatively inexpensive and orally bioavailable anti-inflammatory agents in the future. PMID- 16378500 TI - MAP kinase p38 inhibitors: clinical results and an intimate look at their interactions with p38alpha protein. AB - Mitogen-activated protein kinase p38 is a serine/threonine kinase originally isolated from lipopolysaccharide (LPS) stimulated monocytes. There are four isoforms p38alpha p38beta, p38gamma, and p38delta. The most thoroughly studied isoform is p38alpha, whose activation has been observed in many hematopoietic and non-hematopoietic cell types upon appropriate stimuli. Subsequently, p38alpha kinase has been shown to be involved in the biosynthesis of TNFalpha and IL-1beta at the translational and transcriptional level. MAP kinase p38alpha represents a point of convergence for multiple signaling processes that are activated in inflammation and thus a key potential target for the modulation of cytokine production. The discovery and publication of p38alpha and the pyridinyl-imidazole inhibitor initiated a huge effort by many companies to develop p38alpha inhibitors as potential treatment for inflammatory diseases. Herein we provide a brief overview of recent reported clinical results for AMG 548, BIRB 796, VX 702, SCIO 469, and SCIO 323. However, our focus will be on the binding modes of these inhibitors and other p38 inhibitors in the recent literature. PMID- 16378501 TI - PPARs in diseases: control mechanisms of inflammation. AB - The three isotypes of peroxisome proliferator-activated receptors (PPARs), PPARalpha, beta/delta and gamma, are ligand-inducible transcription factors that belong to the nuclear hormone receptor family. PPARs are implicated in the control of inflammatory responses and in energy homeostasis and thus, can be defined as metabolic and anti-inflammatory transcription factors. They exert their anti-inflammatory effects by inhibiting the induction of pro-inflammatory cytokines, adhesion molecules and extracellular matrix proteins or by stimulating the production of anti-inflammatory molecules. Furthermore, PPARs modulate the proliferation, differentiation and survival of immune cells including macrophages, B cells and T cells. This review discusses the molecular mechanisms by which PPARs and their ligands modulate the inflammatory response. In addition, it presents recent developments implicating PPAR specific ligands in potential treatments of inflammation-related diseases, such as atherosclerosis, inflammatory bowel diseases, Parkinson's and Alzheimer's diseases. PMID- 16378502 TI - Inhibitors of secretory phospholipase A2 group IIA. AB - Phospholipases A2 cleave membrane phospholipids to release arachidonic acid, the precursor to a large family of pro-inflammatory eicosanoids including prostaglandins and leukotrienes that have been proven to exacerbate numerous diseases that have an inflammatory component. Current therapies include NSAIDs' that inhibit cyclooxygenases (COX-1, COX-2) but have no effect on the production of leukotrienes or platelet activating factor (PAF). Inhibitors of PLA2 therefore offer the potential to block production of a more complete set of inflammatory substances through blockade at the onset of the cascade of reactions that follow arachidonic acid release. Many potent, bioavailable and selective inhibitors of human sPLA2 group IIA have been available for more than a decade and have provided compelling support for a causative role of sPLA2 group IIA in numerous studies involving animal models of inflammatory diseases. However, the true value of sPLA2 inhibitors for the treatment of human diseases has had to await phase II clinical trials which have only been completed in the last two years. This review presents the structurally diverse array of available sPLA2 group IIA inhibitors, their associated biological activity in animal models, and evaluation of therapeutic potential in phase II clinical trials in humans. PMID- 16378503 TI - Nonpeptide ligands that target peptide-activated GPCRs in inflammation. AB - The focus of this review is on G protein-coupled receptors (GPCRs) for which nonpeptidic ligands are known and have been evaluated for the treatment of inflammatory conditions. GPCRs are the most prevalent class of cell surface proteins in pharmaceutical research today, and GPCR-targeting drugs account for one tenth of worldwide pharmaceutical sales. Of over 800 human GPCRs identified to date, several hundred are activated by peptides/proteins and just over 30 of these have been identified so far as potential therapeutic targets for the treatment of inflammatory diseases. This review highlights those GPCRs and over 60 structurally diverse nonpeptidic compounds that interact with them and display pro- or anti- inflammatory properties. Among these GPCR targets are the receptors for peptides like bradykinin, chemokines, complement anaphylatoxins, corticotropin releasing factor, endothelins, melanocortins, tachykinins, urocortins, as well as the protease activated receptors (PARs). Other peptide activated GPCRs implicated in inflammation, like those that bind angiotensin II, N-formyl peptides, galanin, neuropeptide Y, opioids and oxytocin, are only briefly discussed because there is either less direct association with inflammation or few/no nonpeptidic antiinflammatory ligands known. While it is still very early in the development of antiinflammatory drugs that target GPCRs, there is already a wealth of information supporting their important roles as cellular sentries in inflammatory diseases. New opportunities are emerging to evaluate antiinflammatory activities of potent and selective GPCR-binding ligands, including those being developed for other disease indications. In summary, GPCRs deserve a great deal more attention as potential therapeutic targets in inflammatory diseases. PMID- 16378505 TI - The nitric oxide signaling pathway in the penis. AB - It is widely accepted that nitric oxide plays an important role in the biology of the penis, serving most familiarly as the agent responsible for penile erection. Early investigation in the field led to the identification of the signaling function of the molecule in the penis which yields corporal smooth muscle relaxation fundamental for the erectile response. Ongoing study of this molecule and its signaling pathway in erectile tissue has served to revise and clarify its importance. Current information conveys the prerequisite of the nitric oxide signaling pathway for penile erection, the regulatory basis for the generation and actions of nitric oxide in the penis, the diverse roles of its synthetic enzyme isoforms in penile biology, and the interaction between nitric oxide and other molecular pathways operative in the broad context of erection physiology. Insight into these subject areas has therapeutic relevance for pathologic conditions of the penis. The purpose of this review is to highlight the latest areas of investigation related to the science of nitric oxide in the penis, as a gateway for considering novel therapeutic strategies for erectile disorders now and in the future. PMID- 16378506 TI - Contractile mechanisms in diabetes-related erectile dysfunction. AB - Penile tumescence (erection) and detumescence (return to the flaccid state) are regulated by a complex neurophysiological process involving the relaxation and contraction, respectively, of smooth muscle (SM) within the two corpus cavernosum (CC) of the penis. Failure of the above SM-mediated process to function properly results in the inability to obtain an erection sufficient for sexual satisfaction and has been termed erectile dysfunction (ED). It is predicted that an estimated 322 million men worldwide will have ED by the year 2025 and, relevant to this review article, is that roughly 50% of men with diabetes also have ED. Furthermore, one of the largest classes of nonresponders to oral phosphodiesterase V (PDE5) inhibitors (the predominant pharmacological treatment for organic ED) are diabetics. This review article examines the current knowledge about the contractile pathways that fine-tune SM tone with particular emphasis on vascular SM including corpus cavernosum smooth muscle (CCSM). The role of the contractile apparatus, SM myosin phosphorylation/dephosphorylation pathways, calcium "sensitization" and "desensitization" pathways and the main neurotransmitters/modulators responsible for regulating CCSM contraction are outlined along with how they are modified or potentially may be modified in response to diabetes. The overall hypothesis generated from this review is that an increased CCSM tone, resulting from an enhancement of contractile mechanisms, may contribute to the higher than average nonresponse rate of diabetic men to PDE5 inhibitors. Knowledge gained from this review will hopefully lead to the generation of drugs that specifically target CCSM contractile pathways which may prove to have therapeutic usefulness in treating ED in diabetics either alone or in combination with existing PDE5 inhibitors. PMID- 16378507 TI - Sonic hedgehog, the penis and erectile dysfunction: a review of sonic hedgehog signaling in the penis. AB - The sinusoid anatomy of the penis is complex and requires complicated interaction between smooth muscle and endothelium in order to maintain homeostasis in the adult. The morphogen, Sonic hedgehog (Shh), is a crucial regulator of these processes, along with its down stream targets patched (Ptc), Hox, bone morphogenetic proteins (BMP's), vascular endothelial growth factor (VEGF) and nitric oxide synthase (NOS). Shh is critical for patterning and establishing tissue identity of the penis during embryonic development, is a crucial regulator of penile postnatal differentiation of the sinusoid morphology of the corpora cavernosa, and plays a fundamental role in maintaining sinusoidal structures pertinent to erectile function in the adult rat. Shh and its targets are active in human penes, and decreased in human diabetic penes in parallel with observations in the rat, thus lending clinical significance to the role of abnormal Shh signaling in erectile dysfunction (ED). Application of exogenous Shh protein to rat corpora cavernosa, induces VEGF and NOS proteins, suggesting a potential mechanism through which decreased Shh protein can cause ED. The studies outlined in this review provide in depth analysis of the Shh pathway and signal transduction, its role in penile development, how Shh signaling is altered in a rat model of ED and neuropathy, how abnormal Shh signaling can cause ED, and the clinical significance of the Shh pathway to human ED. These studies will provide valuable insight, at the molecular level, into understanding the mechanisms that under lie ED and lead to new treatment strategies for diabetic impotence. PMID- 16378508 TI - Rho-kinase and RGS-containing RhoGEFs as molecular targets for the treatment of erectile dysfunction. AB - Erectile dysfunction (ED) is a highly prevalent and often under-treated condition. Erection is basically a spinal reflex that can be initiated by recruitment of penile afferents but also by visual, olfactory and imaginary stimuli. The generated nervous signals will influence the balance between contractile and relaxant factors, which control the degree of contraction of penile corporal cavernosal smooth muscles and, thus, determine the erectile state of the penis. The different steps involved in neurotransmission, impulse propagation and intracellular transduction of neural signals may be changed in different types of ED. Recent studies have revealed important roles for the small GTPase RhoA and its effector, Rho-kinase in regulating cavernosal smooth muscle tone. The RhoA/Rho-kinase pathway modulates the level of phosphorylation of the myosin light chain, mainly through inhibition of myosin phosphatase, and contributes to agonist-induced Ca(2+)-sensitization in smooth muscle contraction. Changes in this pathway may contribute to ED in various patient subgroups (e.g. hypertension, diabetes, hypogonadism). This review summarizes the importance of Rho-kinase signaling in the erectile response and introduces the evidence pointing to RGS-containing Rho-guanine nucleotide exchange factors (GEFs) as critical mediators of RhoA-GTPase activation in cavernosal smooth muscle and its possible compartmentalization in the caveolae. In addition, we suggest that the design of selective inhibitors of these GEFs might represent a novel class of pharmacological agents to treat ED. PMID- 16378509 TI - The pleiotropic effects of inducible nitric oxide synthase (iNOS) on the physiology and pathology of penile erection. AB - The contribution of the neuronal and endothelial isoforms of nitric oxide synthase (nNOS and eNOS, respectively) in the synthesis of nitric oxide as a mediator of penile erection, at the levels of both the penile corpora cavernosa and the hypothalamic regions that control the erectile response, are well established. More recently, the role of the third NOS isoform, the inducible NOS (iNOS), has also started to be elucidated. iNOS does not appear to intervene directly in physiological penile erection or in its central control, but its transcriptional induction is postulated to be a key factor in two opposite related pathological processes, namely neurotoxicity in critical related regions of the hypothalamus during senescence, and as a defense mechanism against the aging or injury-associated fibrosis in the penile corpora cavernosa, the media of the penile arteries, and the tunica albuginea. By counteracting fibrosis that impairs cavernosal smooth muscle compliance, iNOS would protect the erectile tissue. However, further studies are needed to conclusively evaluate these putative roles in the two organs involved in reproductive function. In addition, whether iNOS induction during aging is a major cause in the net loss of trabecular smooth muscle in the corpora cavernosa through apoptosis, remains to be elucidated. The overall evaluation of these conflicting effects is important in order to decide whether pharmacological iNOS induction, or alternatively NO donors or L-arginine, may constitute a valid approach to prevent or treat penile fibrosis and vasculogenic erectile dysfunction. PMID- 16378510 TI - Phosphodiesterase type 5 inhibitors: molecular pharmacology and interactions with other phosphodiesterases. AB - Erectile function is determined by tight regulation of relaxation or contraction of corpus cavernosal smooth muscle, which is the result of a long and complex chain of molecular events. Control of erectile function resides in signaling pathways of the central and peripheral nervous system, as well as intracellular events in the penile smooth muscle. Vascular events resulting in erection have long been understood, and the role of the signaling pathways of the central and peripheral nervous systems in erectile function and dysfunction has become increasingly clear over the last decade. This knowledge has led to the development and current availability of effective oral treatments for erectile dysfunction, the selective phosphodiesterase type 5 (PDE5) inhibitors-sildenafil, vardenafil and tadalafil. In the past few years we have seen an elucidation of the molecular events involved in erectile function and dysfunction and the detailed mechanisms of action by which the specific PDE5 inhibitors work. A review of those mechanisms helps to explain the success of the currently available PDE5 inhibitors and the differences between them and suggests new approaches for developing potential future novel therapies or refinements to existing structures that may improve their efficacy, selectivity and safety profiles. PMID- 16378511 TI - Endothelial nitric oxide synthase gene therapy for erectile dysfunction. AB - Basic science research on erectile physiology has been devoted to investigating the pathogenesis of erectile dysfunction (ED) and has led to the conclusion that ED is predominately a disease of vascular origin. It is well recognized that the incidence of ED dramatically increases in men who suffer from diabetes mellitus, hypercholesterolemia, and cardiovascular disease. Endothelial nitric oxide synthase (eNOS) is an important factor in cardiovascular homeostasis, angiogenesis, and erectile function. Given the impact of endothelial-derived nitric oxide (NO) in vascular biology, a great deal of research over the past decade has focused on the role of NO synthesis from the endothelium in normal erectile physiology as well as in disease states. Loss of the functional integrity of the endothelium and subsequent endothelial dysfunction plays an integral role in the occurrence of ED. Therefore, a likely target of gene therapy for the treatment of ED is eNOS. This communication reviews the role of eNOS in erectile physiology and discusses the alterations in eNOS expression in various vascular diseases of the penis. Putative gene therapy interventions to restore eNOS expression and subsequent endothelial function may represent an exciting new therapeutic strategy for the future treatment of ED. PMID- 16378512 TI - Animal models of ejaculatory behavior. AB - Premature ejaculation is generally regarded the most frequent male ejaculatory complaint and has been considered a psychosexual disorder with psychogenic aetiology. The efficacy of various antidepressants, however, to delay ejaculation in men and to pharmacologically treat premature ejaculation suggests a strong neurobiological involvement. Most of our current understanding of the neurobiology and neuroanatomy of sexual behavior and ejaculatory function has been derived from preclinical studies using several laboratory species. In the present paper we will review the various animal models that have been developed to further study ejaculatory function in the laboratory rat. In addition, we will briefly review the effects of serotonergic antidepressants and serotonergic compounds on sexual and ejaculatory behavior. Together, these preclinical studies may contribute to a better understanding of the neurobiology of ejaculation and help the development of novel drug targets to treat ejaculatory disorders such as premature ejaculation. PMID- 16378513 TI - Magnetic resonance contrast agents: from the bench to the patient. AB - Magnetic Resonance Imaging is gaining a prominent role in the routine clinical investigation. To further improve this technique it is crucial that contrast agents are developed with more optimal organ specificity. This will not only result in a better diagnostic efficiency but also in a reduction of the amount of the agent administered. A combination of techniques has been employed to increase the target selectivity of the contrast agent and thereby the feasibility to visualize different organs. The organ targeting is based on the understanding of the mechanisms involved in the interaction of the agent with plasma proteins (albumin in particular) as well as the different membrane transporters involved in the uptake and in the excretion of the agent from the organ. The physicochemical properties of the contrast agents play a major role in the interaction with these various proteins. In this review we address the relationship between the structure of the contrast agents and their binding to different plasma proteins and membrane transporters in different organs, with special reference to the liver and kidney. The present and potentially future applications of these concepts in the clinical setting are also discussed. PMID- 16378514 TI - Artificial O2 carriers: status in 2005. AB - Donor blood is a limited resource and its transfusion is associated with significant adverse effects. Therefore, alternatives have been searched, the ultimate being artificial oxygen (O2) carriers. There are two main groups of artificial O2 carriers: hemoglobin based and perfluorocarbon emulsions. The hemoglobin molecule in hemoglobin based artificial O2 carriers needs to be stabilized to prevent dissociation of the alpha2beta2-hemoglobin tetramer into alphabeta-dimers in order to prolong intravascular retention and to eliminate nephrotoxicity. Other modifications serve to decrease O2 affinity in order to improve O2 off-loading to tissues. In addition, polyethylene glycol may be surface conjugated to increase molecular size. Finally, certain products are polymerized to increase the hemoglobin concentration at physiologic colloid oncotic pressure. Perfluorocarbons are carbon-fluorine compounds characterized by a high gas dissolving capacity for O2 and CO2 and chemical and biologic inertness. Perfluorocarbons are not miscible with water and therefore need to be brought into emulsion for intravenous application. Development, product specification, physiologic effects, efficacy to decrease the need for donor blood in surgery and side effects of the following products are described: Diaspirin cross-linked hemoglobin (HemAssist), human recombinant hemoglobin (rHb1.1 and rHb2.0), polymerized bovine hemoglobin-based O2 carrier (HBOC-201), human polymerized hemoglobin (PolyHeme), hemoglobin raffimer (Hemolink), maleimide activated polyethylene glycol-modified hemoglobin (MP4) and perflubron emulsion (Oxygent). In addition, enzyme cross-linked poly-hemoglobin, hemoglobin containing vesicles (nano-dimension artificial red blood cells) and an allosteric modifier (RSR13) are discussed. The most advanced products are in clinical phase III trials but no product has achieved market approval yet in the US, Europe or Canada. PMID- 16378515 TI - Polychronization: computation with spikes. AB - We present a minimal spiking network that can polychronize, that is, exhibit reproducible time-locked but not synchronous firing patterns with millisecond precision, as in synfire braids. The network consists of cortical spiking neurons with axonal conduction delays and spike-timing-dependent plasticity (STDP); a ready-to-use MATLAB code is included. It exhibits sleeplike oscillations, gamma (40 Hz) rhythms, conversion of firing rates to spike timings, and other interesting regimes. Due to the interplay between the delays and STDP, the spiking neurons spontaneously self-organize into groups and generate patterns of stereotypical polychronous activity. To our surprise, the number of coexisting polychronous groups far exceeds the number of neurons in the network, resulting in an unprecedented memory capacity of the system. We speculate on the significance of polychrony to the theory of neuronal group selection (TNGS, neural Darwinism), cognitive neural computations, binding and gamma rhythm, mechanisms of attention, and consciousness as "attention to memories." PMID- 16378516 TI - Making working memory work: a computational model of learning in the prefrontal cortex and basal ganglia. AB - The prefrontal cortex has long been thought to subserve both working memory (the holding of information online for processing) and executive functions (deciding how to manipulate working memory and perform processing). Although many computational models of working memory have been developed, the mechanistic basis of executive function remains elusive, often amounting to a homunculus. This article presents an attempt to deconstruct this homunculus through powerful learning mechanisms that allow a computational model of the prefrontal cortex to control both itself and other brain areas in a strategic, task-appropriate manner. These learning mechanisms are based on subcortical structures in the midbrain, basal ganglia, and amygdala, which together form an actor-critic architecture. The critic system learns which prefrontal representations are task relevant and trains the actor, which in turn provides a dynamic gating mechanism for controlling working memory updating. Computationally, the learning mechanism is designed to simultaneously solve the temporal and structural credit assignment problems. The model's performance compares favorably with standard backpropagation-based temporal learning mechanisms on the challenging 1-2-AX working memory task and other benchmark working memory tasks. PMID- 16378518 TI - Oscillatory networks: pattern recognition without a superposition catastrophe. AB - Using an oscillatory network model that combines classical network models with phase dynamics, we demonstrate how the superposition catastrophe of pattern recognition may be avoided in the context of phase models. The model is designed to meet two requirements: on and off states should correspond, respectively, to high and low phase velocities, and patterns should be retrieved in coherent mode. Nonoverlapping patterns can be simultaneously active with mutually different phases. For overlapping patterns, competition can be used to reduce coherence to a subset of patterns. The model thereby solves the superposition problem. PMID- 16378517 TI - Identification of multiple-input systems with highly coupled inputs: application to EMG prediction from multiple intracortical electrodes. AB - A robust identification algorithm has been developed for linear, time-invariant, multiple-input single-output systems, with an emphasis on how this algorithm can be used to estimate the dynamic relationship between a set of neural recordings and related physiological signals. The identification algorithm provides a decomposition of the system output such that each component is uniquely attributable to a specific input signal, and then reduces the complexity of the estimation problem by discarding those input signals that are deemed to be insignificant. Numerical difficulties due to limited input bandwidth and correlations among the inputs are addressed using a robust estimation technique based on singular value decomposition. The algorithm has been evaluated on both simulated and experimental data. The latter involved estimating the relationship between up to 40 simultaneously recorded motor cortical signals and peripheral electromyograms (EMGs) from four upper limb muscles in a freely moving primate. The algorithm performed well in both cases: it provided reliable estimates of the system output and significantly reduced the number of inputs needed for output prediction. For example, although physiological recordings from up to 40 different neuronal signals were available, the input selection algorithm reduced this to 10 neuronal signals that made significant contributions to the recorded EMGs. PMID- 16378519 TI - Topographic product models applied to natural scene statistics. AB - We present an energy-based model that uses a product of generalized Student-t distributions to capture the statistical structure in data sets. This model is inspired by and particularly applicable to "natural" data sets such as images. We begin by providing the mathematical framework, where we discuss complete and overcomplete models and provide algorithms for training these models from data. Using patches of natural scenes, we demonstrate that our approach represents a viable alternative to independent component analysis as an interpretive model of biological visual systems. Although the two approaches are similar in flavor, there are also important differences, particularly when the representations are overcomplete. By constraining the interactions within our model, we are also able to study the topographic organization of Gabor-like receptive fields that our model learns. Finally, we discuss the relation of our new approach to previous work--in particular, gaussian scale mixture models and variants of independent components analysis. PMID- 16378521 TI - Differential log likelihood for evaluating and learning gaussian mixtures. AB - We introduce a new unbiased metric for assessing the quality of density estimation based on gaussian mixtures, called differential log likelihood. As an application, we determine the optimal smoothness and the optimal number of kernels in gaussian mixtures. Furthermore, we suggest a learning strategy for gaussian mixture density estimation and compare its performance with log likelihood maximization for a wide range of real-world data sets. PMID- 16378520 TI - A simple Hebbian/anti-Hebbian network learns the sparse, independent components of natural images. AB - Slightly modified versions of an early Hebbian/anti-Hebbian neural network are shown to be capable of extracting the sparse, independent linear components of a prefiltered natural image set. An explanation for this capability in terms of a coupling between two hypothetical networks is presented. The simple networks presented here provide alternative, biologically plausible mechanisms for sparse, factorial coding in early primate vision. PMID- 16378522 TI - Magnification control in self-organizing maps and neural gas. AB - We consider different ways to control the magnification in self-organizing maps (SOM) and neural gas (NG). Starting from early approaches of magnification control in vector quantization, we then concentrate on different approaches for SOM and NG. We show that three structurally similar approaches can be applied to both algorithms that are localized learning, concave-convex learning, and winner relaxing learning. Thereby, the approach of concave-convex learning in SOM is extended to a more general description, whereas the concave-convex learning for NG is new. In general, the control mechanisms generate only slightly different behavior comparing both neural algorithms. However, we emphasize that the NG results are valid for any data dimension, whereas in the SOM case, the results hold only for the one-dimensional case. PMID- 16378523 TI - Enhancing density-based data reduction using entropy. AB - Data reduction algorithms determine a small data subset from a given large data set. In this article, new types of data reduction criteria, based on the concept of entropy, are first presented. These criteria can evaluate the data reduction performance in a sophisticated and comprehensive way. As a result, new data reduction procedures are developed. Using the newly introduced criteria, the proposed data reduction scheme is shown to be efficient and effective. In addition, an outlier-filtering strategy, which is computationally insignificant, is developed. In some instances, this strategy can substantially improve the performance of supervised data analysis. The proposed procedures are compared with related techniques in two types of application: density estimation and classification. Extensive comparative results are included to corroborate the contributions of the proposed algorithms. PMID- 16378524 TI - Does health care improve health? PMID- 16378525 TI - Improving policy-making in a ministry of health. PMID- 16378527 TI - Quality indicators for cardiac care: national standards in a community context. AB - OBJECTIVE: Public reporting of health data is well established in the United States and in the United Kingdom, and is assumed to promote better health care through informed choice by consumers. To be successful, reporting systems must have the support of physicians, but their opinions have been mixed. The purpose of this study was to explore with practising physicians the perceived usefulness of, and barriers to use of, quality indicators in the care of acute myocardial infarction and congestive heart failure, and the contexts in which these issues arise. METHODS: Six focus groups were conducted in small-, medium- and large sized communities in two provinces in Canada. Subjects were family physicians, emergency physicians, internists and cardiologists. Data were analysed inductively. RESULTS: Our participants were generally supportive of the quality indicators, with concerns expressed regarding interpretation of data from measures created by "experts" but applied in the context of community hospitals and community-based practice. Content analysis disclosed that a majority of the indicators was acceptable; few were outright unacceptable. Inductive analysis revealed two contextual concerns: issues arising from the structure and organization of the health care system, such as equitable access to health care resources and discontinuity or fragmentation of the system, and patient-related issues, such as compliance with medications post-discharge and costs of medications. CONCLUSIONS: There is general support for this set of quality indicators, with the caveat that data should be carefully interpreted in the context of each community in which they are applied. PMID- 16378528 TI - Impact of administrative restrictions on antibiotic use and expenditure in Ontario: time series analysis. AB - OBJECTIVE: In a potential attempt to guide antibiotic prescribing based on current clinical evidence and mitigate the spread of antibiotic resistance, in March 2001 the Ontario Drug Benefit programme restricted reimbursement of two fluoroquinolone antibiotics--ciprofloxacin and ofloxacin--to its beneficiaries. Our objective was to determine the impact of this policy on the volume and cost of antibiotic prescribing. METHOD: Weekly administrative data on antibiotic prescribing volumes and expenditures were analysed between January 1999 and September 2002 to estimate the effect of the policy changes using time series analysis. RESULTS: The policy changes were associated with a statistically significant shift downwards for the fluoroquinolones as a category (1905 fewer prescriptions each week, representing a saving of Can$105,707 a week), driven by a decrease in prescriptions for ciprofloxacin (2084 fewer prescriptions a week, saving Can$129,421 a week). Nitrofurantoin (200 more prescriptions a week, costing an extra Can$2082 a week) and trimethoprim-sulphamethoxazole (532 more prescriptions a week, costing an extra Can$1473 a week) demonstrated a statistically significant shift upwards. The latter also showed a decrease in trend and nitrofurantoin an increase in trend during the time period. There was no statistically significant change in either the total number of antibiotic prescriptions or expenditures associated with the policy of limiting their use. CONCLUSIONS: Although no direct cause and effect can be shown with these observational data, the results suggest that the change in reimbursement policy to restrict prescribing of fluoroquinolones decreased their use and associated expenditures. These decreases were offset by increases in the use of other antibiotics. The balance of consequent benefit and harm of these shifts in prescribing patterns needs to be examined carefully. Alternative solutions to encourage appropriate use of antibiotics deserve exploration. PMID- 16378529 TI - Do the incentive payments in the new NHS contract for primary care reflect likely population health gains? AB - OBJECTIVE: The new contract for primary care in the UK offers fee-for-service (FFS) payments for a wide range of activities in a quality outcomes framework (QOF), with payments designed to reflect likely workload. This study aims to explore the link between these financial incentives and the likely population health gains. METHODS: The study examines a subset of eight preventive interventions covering 38 of the 81 clinical indicators in the quality framework. The maximum payment for each service was calculated and compared with the likely population health gain in terms of lives saved per 100,000 population based on evidence from McColl et al. (1998). RESULTS: Maximum payments for the eight interventions examined make up 57% of the total maximum payment for all clinical interventions in the (QOF). There appears to be no relationship between pay and health gain across these eight interventions. Two of the eight interventions (warfarin in atrial fibrillation and statins in primary prevention) receive no incentive. CONCLUSIONS: Payments in the new contract do not reflect likely population health gain. There is a danger that clinical activity may be skewed towards high-workload activities that are only marginally effective, to the detriment of more cost-effective activities. If improving population health is the primary goal of the NHS, then FFS incentives should be designed to reflect likely health gain rather than likely workload. PMID- 16378530 TI - Ethics and economics: does programme budgeting and marginal analysis contribute to fair priority setting? AB - OBJECTIVE: Limited resources mean that decision-makers must set priorities among competing opportunities. Programme budgeting and marginal analysis (PBMA) is an economic approach that focuses on optimizing benefits with available resources. Accountability for reasonableness (A4R) is an ethics approach that focuses on ensuring fair priority-setting processes. PBMA and A4R have been used separately to provide decision-makers with advice about how to set priorities within limited resources. The goals of this research were to use the A4R framework to evaluate the fairness of using PBMA for priority setting and to assess how A4R might make PBMA fairer. METHODS: Qualitative case studies to describe priority setting using PBMA in the Calgary Health Region (Alberta, Canada) evaluated using A4R as a conceptual framework. RESULTS: The use of PBMA for priority setting was fairer than previous priority setting because of its emphasis on explicit rational decision-making. However, there were opportunities to improve the process, particularly by collecting data related to the decision criteria, by developing a communication plan to engage internal and external stakeholders about priority setting, and by providing a formal mechanism to review priority-setting decisions and resolve disputes. CONCLUSIONS: There is potential for combining A4R and PBMA in a more comprehensive approach to priority setting, which uses a fair priority setting process to reach decisions aimed at achieving optimal benefits with available resources. PMID- 16378531 TI - Q methodology in health economics. AB - The recognition that health economists need to understand the meaning of data if they are to adequately understand research findings which challenge conventional economic theory has led to the growth of qualitative modes of enquiry in health economics. The use of qualitative methods of exploration and description alongside quantitative techniques gives rise to a number of epistemological, ontological and methodological challenges: difficulties in accounting for subjectivity in choices, the need for rigour and transparency in method, and problems of disciplinary acceptability to health economists. Q methodology is introduced as a means of overcoming some of these challenges. We argue that Q offers a means of exploring subjectivity, beliefs and values while retaining the transparency, rigour and mathematical underpinnings of quantitative techniques. The various stages of Q methodological enquiry are outlined alongside potential areas of application in health economics, before discussing the strengths and limitations of the approach. We conclude that Q methodology is a useful addition to economists' methodological armoury and one that merits further consideration and evaluation in the study of health services. PMID- 16378532 TI - The biggest bang for the buck or bigger bucks for the bang: the fallacy of the cost-effectiveness threshold. AB - It has been suggested that scepticism among decision-makers about using cost effectiveness analysis (CEA) is caused in part by the low level of the cost effectiveness "thresholds" in the economic evaluation literature. This has led Ubel and colleagues to call for higher threshold values of US$200,000 or more per quality-adjusted life-year. We show that these arguments fail to identify the objective of CEA and hence do not consider whether or how the threshold relates to this objective. We show that incremental cost-effectiveness ratios (ICERs) cannot be used to identify an efficient use of resources--the "biggest bang for the bucks"--allocated to health care. On the contrary, the practical consequence of using the ICER approach is shown to be an increase in health care expenditures, or "bigger bucks for making a bang", without any evidence of the bang being bigger (i.e. that this leads to an increase in benefits to the population). We present an alternative approach that provides an unambiguous method of determining whether a new intervention leads to an increase in health gains from whatever resources are to be made available to health care decision makers. PMID- 16378533 TI - All dressed up but nowhere to go? Delayed hospital discharges and older people. AB - OBJECTIVE: Delayed hospital discharges are a key concern in a number of industrialized nations and are the subject of a range of government initiatives in the English National Health Service. The aim of this paper was to review the UK literature on delayed hospital discharges and older people in order to identify and explore the rate and causes of delayed hospital discharges, together with policies and practices that may reduce delayed discharges and improve the experiences of older people. METHODS: Literature review based on searches of major health/social-care databases. Sources which explore the rate and cause of delayed discharges in the UK were included. Relevant documents were categorized using the research hierarchy set out in the National Service Framework for Older People and analysed according to criteria for appraising the quality of qualitative research proposed by Mays et al. RESULTS: The review identified 21 studies, which suggest very different rates and causes of delayed discharge in different settings. The studies reveal the importance of rehabilitation services to reduce the rate of delayed discharge, the prevalence of delayed discharges caused by internal hospital factors, and the complex and multi-faceted nature of the factors contributing to delayed discharge. Despite this, the studies have a number of methodological flaws and often fail to include a patient perspective or to consider detailed policies and approaches to reduce the number of delayed discharges. There is also a failure to consider the needs of older people with mental health problems or people from minority ethnic communities. CONCLUSION: The evidence, as it currently stands, raises a number of issues about current hospital discharge policy, supporting some aspects of the current government agenda in England, but questioning other aspects. PMID- 16378536 TI - We can eliminate errors in health care by getting rid of the "bad apples". PMID- 16378556 TI - The White Paper: what the College wants. PMID- 16378557 TI - Developing an evidence base for intermediate care delivered by GPs with a special interest. PMID- 16378558 TI - Triage and remote consultations: moving beyond the rhetoric of access and choice. PMID- 16378559 TI - Randomised controlled trial to compare GP-run orthopaedic clinics based in hospital outpatient departments and general practices. AB - BACKGROUND: To reduce outpatient waiting times, a growing number of outpatient clinics for selected groups of patients are being provided by GPs with special interests (GPwSIs). AIM: To determine whether there are differences in patient satisfaction or clinical outcome among patients attending orthopaedic clinics provided by GPwSIs in hospital or community settings. DESIGN OF STUDY: Randomised controlled trial. SETTING: Hospital outpatient departments or general practices. METHOD: Three hundred and twenty-one patients with minor orthopaedic problems were referred by GPs to the orthopaedic surgery department of the University Hospitals of Leicester NHS Trust; 168 patients were randomised to care by GPwSIs in practices, and 153 were randomised to care by the same GPwSIs in clinics held at hospital outpatient departments. Patients completed the SF-36v2 and satisfaction questionnaires at their first appointment, and again 3 months later. RESULTS: There was no significant difference between the sites in changes in health. After the first clinic attendance, patients attending practice-based clinics were more satisfied with access to appointments and information received. CONCLUSION: For selected orthopaedic referrals seen by GPwSIs, there were no significant differences in clinical outcomes between practice-based and hospital based clinics, but some features of practice-based clinics tend to be preferred by patients. PMID- 16378560 TI - Targeted routine asthma care in general practice using telephone triage. AB - BACKGROUND: There is a high non-attendance rate for traditional clinic-based routine asthma care in general practice. Alternative methods of providing routine asthma care need to be examined. AIM: To examine the cost and effectiveness of targeted routine asthma care in general practice using telephone triage, compared to usual clinic care. DESIGN OF STUDY: An open randomised controlled trial. SETTING: A single semi-rural practice in the southwest of England. METHOD: Adult patients with asthma were randomised to receive either their routine asthma care in the surgery or care by telephone triage. Asthma control parameters, health status and NHS resource utilisation were measured over the 12-month study period. RESULTS: One hundred and ninety-four patients were randomised and 35% per cent more patients (n = 84 versus n = 62) received more than one consultation in the telephone group. Asthma control as measured by the asthma control questionnaire (ACQ) was similar in the clinic and telephone groups: mean change in ACQ = -0.11 (95% CI = -0.32 to 0.11) versus -0.18 (95% CI = -0.38 to 0.02). Mean NHS costs were 210 pounds sterling per patient per year in the telephone group compared to 334 pounds sterling in the clinic group (P-value of bootstrapped difference = 0.071). CONCLUSION: Targeted routine asthma care by telephone triage of adult asthmatics can lead to more asthma patients being reviewed, at less cost per patient and without loss of asthma control compared to usual routine care in the surgery. PMID- 16378561 TI - The treatment of acute infectious conjunctivitis with fusidic acid: a randomised controlled trial. AB - BACKGROUND: Acute infectious conjunctivitis is a common disorder in primary care. Despite a lack of evidence regarding the effectiveness of topical antibiotics for the treatment of acute infectious conjunctivitis, most patients presenting in primary care with the condition receive topical antibiotics. In The Netherlands, fusidic acid is most frequently prescribed. AIM: To assess the effectiveness of fusidic acid gel compared to placebo for acute infectious conjunctivitis. DESIGN: Double-blind randomised placebo-controlled trial. SETTING: Twenty-five Dutch primary care centres. METHOD: Adults presenting with a red eye and either (muco)purulent discharge or glued eyelid(s) were allocated to either one drop of fusidic acid gel 1% or placebo, four times daily during one week. The main outcome measure was the difference in recovery rates at 7 days. Secondary outcome measures were difference in bacterial eradication rates, a survival time analysis of the duration of symptoms, and the difference in recovery rates in culture positive and culture-negative patients. RESULTS: One hundred and eighty-one patients were randomised and 163 patients were analysed. Forty-five of the 73 patients in the treatment and 53 of the 90 patients in the placebo group recovered (adjusted risk difference = 5.3% [95% confidence interval {CI} = -11 to 18]). There was no difference between the median duration of symptoms in the two groups. At baseline, the prevalence of a positive bacterial culture was 32% (58/181). The bacterial eradication rate was 76% in the treatment and 41% in the placebo group (risk difference = 35% [95% CI = 9.3 to 60.4]). In culture positive patients, the treatment effect tended to be strong (adjusted risk difference = 23% [95% CI = -6 to 42]). CONCLUSION: At 7 days, cure rates in the fusidic acid gel and placebo group were similar, but the confidence interval was too wide to clearly demonstrate their equivalence. These findings do not support the current prescription practices of fusidic acid by GPs. PMID- 16378562 TI - Organisational factors in relation to control of blood pressure: an observational study. AB - BACKGROUND: Studies show that 60-75% of treated patients with hypertension in general practice, still do not reach the recommended blood pressure targets of <150/90 mmHg. AIM: To investigate aspects of hypertension management in relation to sociodemographic variables, antihypertensive drug treatment, and organisational factors in primary care. DESIGN OF STUDY: Observational study over 3 years. SETTING: Eight general practices in Tayside, UK. METHOD: Participants were 560 randomly selected patients aged 40-79 years receiving treatment for hypertension. The outcome measurement was blood pressure control, expressed in binary form based on the British Hypertension Society audit standard of <150/90 mmHg. RESULTS: Of 536 eligible patients, 261 (49%) were defined as having inadequate blood pressure control at the end of the study period. No significant associations were discovered with sex, age, deprivation score and comorbidity. In those patients with inadequate control, 30% had no modifications to their drug treatment during the study period. Blood pressure control at the end of the study period was not associated with number of antihypertensive drugs taken or number of antihypertensive drug modifications. The mean number of clinician contacts was 11 (standard deviation = 8), and mean continuity in primary care was high, although this was not associated with improved blood pressure control. A higher proportion of hypertension-related consultations were associated with increased odds of having inadequate blood pressure control. CONCLUSION: Achieving adequate blood pressure control continues to represent a substantial health problem in a significant proportion of the hypertensive population. Patient, physician and organisational elements play a role in ensuring effective delivery of hypertension care in the community. PMID- 16378563 TI - Comparison of GP and nurse practitioner consultations: an observational study. AB - BACKGROUND: Studies show that satisfaction with nurse practitioner care is high when compared with GPs. Clinical outcomes are similar. Nurse practitioners spend significantly longer on consultations. AIM: We aimed to discover what nurse practitioners do with the extra time, and how their consultations differ from those of GPs. DESIGN OF STUDY: Comparative content analysis of audiotape transcriptions of 18 matched pairs of nurse practitioner and GP consultations. SETTING: Nine general practices in south Wales and south west England. METHOD: Consultations were taped and clinicians' utterances coded into categories developed inductively from the data, and deductively from the literature review. RESULTS: Nurse practitioners spent twice as long with their patients and both patients and clinicians spoke more in nurse consultations. Nurses talked significantly more than GPs about treatments and, within this, talked significantly more about how to apply or carry out treatments. Weaker evidence was found for differences in the direction of nurses being more likely to: discuss social and emotional aspects of patients' lives; discuss the likely course of the patient's condition and side effects of treatments; and to use humour. Some of the extra time was also spent in getting doctors to approve treatment plans and sign prescriptions. CONCLUSIONS: The provision of more information in the longer nurse consultations may explain differences in patient satisfaction found in other studies. Clinicians need to consider how much information it is appropriate to provide to particular patients. PMID- 16378564 TI - Estimation of blood glucose levels by people with diabetes: a cross-sectional study. AB - BACKGROUND: Recommendations suggest that all patients with diabetes who use insulin should home test their blood glucose. Recommendations for those not using insulin remain contradictory. These recommendations are in part based upon the assumption that people with diabetes cannot make an accurate estimate. AIM: To explore whether people with diabetes can accurately estimate their blood glucose levels and to assess which factors explain variability in these estimates. DESIGN: A cross-sectional design. SETTING: One general practice in Oxfordshire, UK. PARTICIPANTS: One hundred and fifteen consecutive patients with diabetes attending a diabetic clinic were invited to estimate their blood glucose level prior to having it routinely measured. RESULTS: One hundred and four patients made estimates. Of these, 45 (43.3%) underestimated their blood glucose, 18 (17.3%) overestimated, and 41 (39.4%) made guesses that fell into the range defined as accurate. Of those not using insulin (n = 85), 37 (43.5%) underestimated their blood glucose, 12 (14%) overestimated and 36 (42.3%) were accurate. Accuracy in the non-insulin users was associated with home testing, lower blood glucose levels, coming to the clinic in a fasting state, and reporting no symptoms when they felt that their blood glucose level was high. Overestimation was associated with having co-occurring illnesses and experiencing no symptoms when their blood glucose was low. CONCLUSION: The majority of patients with diabetes in this study could not accurately estimate their blood glucose levels indicating that home testing may be a necessary part of diabetes self care. Home testing may also function as a form of biofeedback to facilitate an improved ability to estimate blood glucose levels. PMID- 16378565 TI - Factors identifying higher risk rectal bleeding in general practice. AB - BACKGROUND: Rectal bleeding is a common symptom. The ability to distinguish those patients having serious underlying pathology from those with self-limiting conditions is a continuing dilemma in general practice. AIM: To determine the factors affecting the predictive and diagnostic value of rectal bleeding for bowel cancer in primary care. DESIGN OF STUDY: One-year prospective observational study. SETTING: Three large general practices. METHOD: Three hundred and nineteen consecutive patients over the age of 34 years consulting their GPs with rectal bleeding were included in the study. Investigation was by flexible sigmoidoscopy or a questionnaire and review of all patients took place after 18 months. The main outcome measures were consultation rates; the prevalence of cancer, colitis and significant polyps in patients presenting with rectal bleeding; its diagnostic value when occurring with or without a change in bowel habit, perianal symptoms and abdominal pain. RESULTS: The consultation rate for rectal bleeding in patients over the age of 34 years was 15 per 1000 per year; 3.4% had colorectal cancer. The prevalence of cancer increased to 9.2% when the rectal bleeding was associated with a change in bowel habit, and to 11.1% when it was without perianal symptoms. Thirty-six per cent of cancer patients had a palpable rectal mass. CONCLUSION: Over 96% of the patients who present to their GPs with rectal bleeding do not have cancer. Greater awareness of the diagnostic value of the different symptom combinations of rectal bleeding could help GPs adopt different management strategies for patients at higher and very low risk of cancer. PMID- 16378566 TI - The effects of telephone consultation and triage on healthcare use and patient satisfaction: a systematic review. AB - BACKGROUND: In recent years there has been a growth in the use of the telephone consultation for healthcare problems. This has developed, in part, as a response to increased demand for GP and accident and emergency department care. AIM: To assess the effects of telephone consultation and triage on safety, service use, and patient satisfaction. DESIGN OF STUDY: We looked at randomised controlled trials, controlled studies, controlled before/after studies, and interrupted time series of telephone consultation or triage in a general healthcare setting. SETTING: All healthcare settings were included but the majority of studies were in primary care. METHOD: We searched the Cochrane Central Register of Controlled Trials, EPOC specialised register, PubMed, EMBASE, CINAHL, SIGLE, and the National Research Register and checked reference lists of identified studies and review articles. Two reviewers independently screened studies for inclusion, extracted data, and assessed study quality. RESULTS: Nine studies met our inclusion criteria: five randomised controlled trials; one controlled trial; and three interrupted time series. Six studies compared telephone consultation with normal care; four by a doctor, one by a nurse, and one by a clinic clerk. Three of five studies found a significant decrease in visits to GPs but two found an increase in return consultations. In general at least 50% (range = 25.5-72.2%) of calls were handled by telephone consultation alone. Of seven studies reporting accident and emergency department visits, six showed no difference between the groups and one--of nurse telephone consultation--found an increase. Two studies reported deaths and found no difference between nurse telephone consultation and normal care. CONCLUSIONS: Although telephone consultation appears to have the potential to reduce GP workload, questions remain about its effect on service use. Further rigorous evaluation is needed with emphasis on service use, safety, cost, and patient satisfaction. PMID- 16378567 TI - Topical antibiotics for acute bacterial conjunctivitis: Cochrane systematic review and meta-analysis update. AB - BACKGROUND: Uncertainty remains about the extent to which findings from our previously published systematic review and meta-analysis of double-blind, randomised controlled trials of topical antibiotics compared with placebo in the management of patients with acute bacterial conjunctivitis treated in secondary care outpatient settings are generalisable to the management of the condition in primary care settings. We updated our review, undertaking searches, methodological assessment, data extraction and analysis according to a pre defined protocol. In addition to the previous three included studies, we identified two additional double-blind primary care trials, one which compares fusidic acid gel with placebo gel and one which compares chloramphenicol eye drops with placebo eye drops in children. Meta-analyses of clinical and microbiological remission data reveal that topical antibiotics are of benefit in improving early (days 2-5) clinical (relative risk [RR] = 1.24, 95% confidence interval [CI] = 1.05 to 1.45) and microbiological (RR = 1.77, 95% CI = 1.23 to 2.54) remission rates; later (days 6-10) data reveal that these early advantages in clinical (RR = 1.11, 95% CI = 1.02 to 1.21) and microbiological cure rates are reduced (RR 1.56, 95% CI = 1.17 to 2.09), but persist. Most cases of acute bacterial conjunctivitis resolve spontaneously. While topical antibiotics are associated with significantly improved rates of early (days 2-5) clinical remission, this benefit is marginal for later remission (days 6-10). PMID- 16378568 TI - Referrals for cataract and Action on cataracts, evidence-based guidelines. PMID- 16378569 TI - The challenge of recruiting people with schizophrenia to trials. PMID- 16378570 TI - Continuing care. PMID- 16378571 TI - Should GPs be prescribing more vitamin D? PMID- 16378572 TI - Impact of obesity. PMID- 16378573 TI - Mental health screening may prove effective in primary care. PMID- 16378574 TI - Prescribing to substance misusers. PMID- 16378575 TI - Assisted suicide. PMID- 16378576 TI - The law on assisted dying. PMID- 16378580 TI - Evidence in practice--number 8. What is the prognosis of optic neuritis? How often does it lead to multiple sclerosis? PMID- 16378582 TI - Perceptual capacity and the good GP: invisible, yet indispensable for quality of care. PMID- 16378590 TI - Ectopic expression of a polyalanine expansion mutant of poly(A)-binding protein N1 in muscle cells in culture inhibits myogenesis. AB - Oculopharyngeal muscular dystrophy (OPMD) is an adult-onset dominant genetic disease caused by the expansion of a GCG trinucleotide repeat that encodes the polyalanine tract at the N-terminus of the nuclear poly(A)-binding protein (PABPN1). Presence of intranuclear inclusions (INIs) containing PABPN1 aggregates in the skeletal muscles is the hallmark of OPMD. Here, we show that ectopic expression of the mutant PABPN1 produced INIs in a muscle cell culture model and reduced expression of several muscle-specific proteins including alpha-actin, slow troponin C, muscle creatine kinase, and two myogenic transcription factors, myogenin and MyoD. However, the levels of two upstream regulators of the MyoD gene, the Myf-5 and Pax3/7, were not affected, but both proteins co-localized with the PABPN1 aggregates in the mutant PABPN1 overexpressing cells. In these cells, although myogenin and MyoD levels were reduced, these two transcription factors did not co-localize with the mutant PABPN1 aggregates. Therefore, sequestration of Myf5 and Pax3/7 by the mutant PABPN1 aggregates was a specific effect on these factors. Our results suggest that trapping of these two important myogenic determinants may interfere with an early step in myogenesis. PMID- 16378591 TI - Caenorhabditis elegans WASP-interacting protein homologue WIP-1 is involved in morphogenesis through maintenance of WSP-1 protein levels. AB - Mammalian WASP and N-WASP are involved in reorganization of the actin cytoskeleton through activation of the Arp2/3 complex and in regulation of cell motility or cell shape changes. In the present study, we identified WASP interacting protein homologue (WIP)-1 in Caenorhabditis elegans. WIP-1 contains the domains and sequences conserved among mammalian WIP family proteins. Yeast two-hybrid analysis detected a physical interaction between WIP-1 and WSP-1, the sole homologue of WASP/N-WASP in C. elegans. Western analysis of embryo lysates showed that RNA interference (RNAi) treatment for wip-1 decreased levels of WSP-1 protein, and wsp-1(RNAi) treatment decreased levels of WIP-1 protein. However, wsp-1 mRNA levels were not decreased in wip-1(RNAi)-treated embryos, and wip-1 mRNA levels were not decreased in wsp-1(RNAi)-treated embryos. Furthermore, disruption of WIP-1 by RNAi resulted in embryonic lethality with morphologic defects in hypodermal cell migration, a process known as ventral enclosure. This phenotype was similar to that observed in RNAi experiments for wsp-1. Immunostaining showed that WIP-1 was expressed by migrating hypodermal cells, as was WSP-1. This expression during ventral enclosure was reduced in wip-1(RNAi) treated embryos and wsp-1(RNAi)-treated embryos. Our results suggest that C. elegans WIP-1 may function in hypodermal cell migration during ventral enclosure by maintaining levels of WSP-1. PMID- 16378592 TI - Molecular identification of the first SIFamide receptor. AB - SIFamide is the short name and also the C terminus of the Drosophila neuropeptide AYRKPPFNGSIFamide. SIFamide has been isolated or predicted from various insects and crustaceans, and appears to be extremely well conserved among these arthropods. However, the function of this neuropeptide is still enigmatic. Here, we have identified the Drosophila gene (CG10823) coding for the SIFamide receptor. When expressed in Chinese hamster ovary cells, the receptor is only activated by Drosophila SIFamide (EC(50), 2x10(-8)M) and not by a library of 32 other insect neuropeptides and eight biogenic amines. Database searches revealed SIFamide receptor orthologues in the genomes from the malaria mosquito Anopheles gambiae, the silkworm Bombyx mori, the red flour beetle Tribolium castaneum, and the honey bee Apis mellifera. An alignment of the five insect SIFamide or SIFamide-like receptors showed, again, an impressive sequence conservation (67 77% amino acid sequence identities between the seven-transmembrane areas; 82-87% sequence similarities). The identification of well-conserved SIFamide receptor orthologues in all other insects with a sequenced genome, suggests that the SIFamide/receptor couple must have an essential function in arthropods. This paper is the first report on the identification of a SIFamide receptor. PMID- 16378593 TI - In vitro expression of natriuretic peptides in cardiomyocytes differentiated from monkey embryonic stem cells. AB - Functional characterization of ES cell-derived cardiomyocytes is important for differentiation control and application to the cell therapy. One of the crucial functions of cardiomyocytes is a production of atrial and brain natriuretic peptides (ANP and BNP, respectively), which have important endocrine, autocrine, and paracrine functions. In this study, we focused on the functional aspect of the cardiomyocytes differentiated from monkey ES cells in vitro and investigated the expression of ANP and BNP. Spontaneously contracting cells showed nodal-like action potentials, and expression of ANP and BNP by RT-PCR and immunocytochemistry. Interestingly, ANP and BNP expressions were detected as immunoreactive granules in the perinuclear area and these signals appeared to co localize with trans-Golgi network. These findings suggest that monkey ES cells were able to differentiate into cardiomyocytes with functional characteristics in vitro and therefore can be used as a useful model to study mechanisms and functions in early cardiogenesis. PMID- 16378594 TI - Enhanced phosphorylation of cyclic AMP response element binding protein in the brain of mice following repetitive hypoxic exposure. AB - Cerebral ischemic/hypoxic preconditioning (I/HPC) is a phenomenon of endogenous protection that renders the brain tolerant to sustained ischemia/hypoxia. This profound protection induced by I/HPC makes it an attractive target for developing potential clinical therapeutic approaches. However, the molecular mechanism of I/HPC is unclear. Cyclic AMP (cAMP) response element binding protein (CREB), a selective nuclear transcriptional factor, plays a key role in the neuronal functions. Phosphorylation of CREB on Ser-133 may facilitate its transcriptional activity in response to various stresses. In the current study, we observed the changes in CREB phosphorylation (Ser-133) and protein expression in the brain of auto-hypoxia-induced HPC mice by using Western blot analysis. We found that the levels of phosphorylated CREB (Ser-133), but not protein expression of CREB, increased significantly (p<0.05) in the hippocampus and the frontal cortex of mice after repetitive hypoxic exposure (H2-H4, n=6 for each group), when compared to that of the normoxic (H0, n=6) or hypoxic exposure once group (H1, n=6). In addition, a significant enhancement (p<0.05) of CREB phosphorylation (Ser-133) could also be found in the nuclear extracts from the whole hippocampus of hypoxic preconditioned mice (H2-H4, n=6 for each group). These results suggest that the phosphorylation of CREB might be involved in the development of cerebral hypoxic preconditioning. PMID- 16378595 TI - Isoform specific changes in PPAR alpha and beta in colon and breast cancer with differentiation. AB - To investigate the role of peroxisome proliferator-activated receptors (PPARs) alpha and beta in the differentiation of colon cancer cells, we differentiated HT 29 cells using sodium butyrate (NaB) and culturing post-confluence and assessed differentiation using the marker intestinal alkaline phosphatase. While PPARalpha levels only changed with culturing post confluence, PPARbeta levels increased independent of the method of differentiation. To explore further the differences induced by NaB, we assessed changes in both PPAR isoforms in MCF-7 breast cancer cells cultured in the presence of NaB over 48h. Again a very different expression pattern was observed with PPARalpha increasing after 4h and remaining elevated, while PPARbeta increased transiently. Our studies suggest that the expression of PPARs is dependent upon both the method of differentiation and on time. Moreover, these studies show that changes in PPARalpha levels are not required for the differentiation of colon cancer cell lines, whereas changes in PPARbeta are more closely associated with differentiation. PMID- 16378596 TI - In vitro cardiomyogenic potential of human umbilical vein-derived mesenchymal stem cells. AB - Cardiomyocyte loss in the ischemically injured human heart often leads to irreversible defects in cardiac function. Recently, cellular cardiomyoplasty with mesenchymal stem cells, which are multipotent cells with the ability to differentiate into specialized cells under appropriate stimuli, has emerged as a new approach for repairing damaged myocardium. In the present study, the potential of human umbilical cord-derived mesenchymal stem cells to differentiate into cells with characteristics of cardiomyocyte was investigated. Mesenchymal stem cells were isolated from endothelial/subendothelial layers of the human umbilical cords using a method similar to that of human umbilical vein endothelial cell isolation. Isolated cells were characterized by transdifferentiation ability to adipocytes and osteoblasts, and also with flow cytometry analysis. After treatment with 5-azacytidine, the human umbilical cord derived mesenchymal stem cells were morphologically transformed into cardiomyocyte-like cells and expressed cardiac differentiation markers. During the differentiation, cells were monitored by a phase contrast microscope and their morphological changes were demonstrated. Immunostaining of the differentiated cells for sarcomeric myosin (MF20), desmin, cardiac troponin I, and sarcomeric alpha-actinin was positive. RT-PCR analysis showed that these differentiated cells express cardiac-specific genes. Transmission electron microscopy revealed a cardiomyocyte-like ultrastructure and typical sarcomers. These observations confirm that human umbilical cord-derived mesenchymal stem cells can be chemically transformed into cardiomyocytes and can be considered as a source of cells for cellular cardiomyoplasty. PMID- 16378597 TI - Stratum intermedium lineage diverges from ameloblast lineage via Notch signaling. AB - The stratum intermedium develops as flattened cell layers on the proximal side of the ameloblast layer during tooth development. However, little information is available regarding the origin and the role. In this study, we indicate that some stratum intermedium cells originate from the inner enamel epithelium (IEE) in rat incisor organ cultures using DiI as a tracer. Immunohistochemical and in situ hybridization studies showed that the stratum intermedium cells express the Notch1 protein and Hes1 mRNAs, while the IEE and ameloblasts express the Jagged1. Further, we examined the role of Notch signaling using the dental epithelial cell line HAT-7. Recombinant Jagged1 protein enhanced the appearance of stratum intermedium cells in HAT-7 cultures and neutralization with an anti-Jagged1 antibody inhibited these effects. Additionally, overexpression of the Notch1 internal domain increased the number of stratum intermedium cells. We hypothesize that the stratum intermedium lineage differentiates from the ameloblast lineage via Notch signaling. PMID- 16378598 TI - Inhibition of the PI3 kinase/Akt pathway enhances doxorubicin-induced apoptotic cell death in tumor cells in a p53-dependent manner. AB - Constitutive activation of the PI3 kinase/Akt pathway is associated with the neoplastic phenotype of a large number of human tumor cells. As the anti apoptotic role of the PI3 kinase/Akt pathway has been established, we have examined whether specific blockade of this pathway sensitizes tumor cells to DNA damaging agent-induced cytotoxicity by enhancing apoptotic cell death. Although a PI3 kinase inhibitor, LY294002, by itself does not induce apoptotic cell death, LY294002 selectively and markedly enhances the apoptosis-inducing efficacy of doxorubicin: such an enhanced cell death is only detected in tumor cells in which the PI3 kinase/Akt pathway is constitutively activated, and it is totally dependent on the functional p53 pathway. These results suggest that the combination of a PI3 kinase/Akt pathway inhibitor and doxorubicin provides an efficient chemotherapeutic strategy for the treatment of tumor cells in which the PI3 kinase/Akt pathway is constitutively activated and the p53 pathway is functional. PMID- 16378599 TI - Identification of the proteins specifically binding to the rat LINE1 promoter. AB - The initial step of LINE1 retrotransposons dissemination requires transcription from species-specific promoter located within 5'-untranslated region of LINE1. Although the 5'-untranslated region of the rat LINE1 element shows promoter activity, no promoter-binding proteins have been discovered so far. Using an EMSA and Southwestern blotting methods, we identified Sp1 and Sp3 proteins, which specifically bind to the rat LINE1 promoter in vitro. The Sp1/Sp3-binding motif within rat LINE1 promoter is located downstream of the major predicted transcription initiation site. PMID- 16378600 TI - Human CC chemokine CCL23 enhances expression of matrix metalloproteinase-2 and invasion of vascular endothelial cells. AB - Human CCL23 (also known as CKbeta8, MPIF-1, or MIP-3) has been recently reported to induce endothelial cell migration and tube formation via CCR1. Matrix metalloproteinases (MMPs) are involved in the degradation of the extracellular matrix and also appear to play critical roles in angiogenesis. In the present study, we have demonstrated that CCL23 enhances the expression of MMP-2 mRNA and protein levels in endothelial cells in a dose-dependent manner, but has no effect on the expression levels of MMP-9, TIMP-1, TIMP-2, and MT1-MMP. CCL23 was shown to dose-dependently activate the expression of the MMP-2/Luc reporter gene, thereby indicating that it stimulates the transcription of the MMP-2 gene. Vascular endothelial cells, when exposed to CCL23, showed a marked ability to invade through a 3D Matrigel. This increase in invasion was also correlated with enhancements in the expression and activity of MMP-2. Neutralization with anti CCL23 and anti-CCR1 antibodies, as well as the heat-induced inactivation of CCL23, resulted in a blockage of the CCL23-activated invasion, indicating that the invasion of HUVECs was induced by CCL23 specifically. Furthermore, we showed that the CCL23-induced invasion was inhibited by MMP inhibitors such as GM6001 and a specific MMP-2 Inhibitor I. Our results indicate that CCL23 may play a direct role in angiogenesis, via the upregulation of MMP-2 expression. PMID- 16378601 TI - 3'-UTR polymorphism in the human CYP2A6 gene affects mRNA stability and enzyme expression. AB - Cytochrome P450 2A6 (CYP2A6) is the major nicotine C-oxidase in human and participates in the metabolism of drugs and precarcinogens. The CYP2A6 gene is highly polymorphic and more than 22 different alleles have been described. We here focused on the polymorphism in the 3'-UTR region, in particular the common CYP2A6*1B allele, carrying an unequal crossover element from the pseudogene CYP2A7. Analysis of CYP2A6 expression in a human liver bank (n=46) revealed that the protein level and catalytic activity using coumarin as a substrate were all higher, following a linear gene-dose relationship, in livers carrying one or two copies of CYP2A6*1B, as compared to other CYP2A6 allelic variants. Different variants of the CYP2A6 3'-UTR were cloned into a modified pGL3 plasmid downstream of the luciferase reporter gene. The plasmids, having the proximal promoter of CYP2A6 gene, were transfected into HeLa cells or injected into the tail veins of male CD1 mice. In both systems, the 3'-UTR CYP2A6*1B constructs caused higher reporter gene activity and the CYP2A7 3'-UTR construct lower activity, compared to the CYP2A6*1 3'-UTR constructs. Two SNPs differentiating the 3'-UTR between CYP2A7 and CYP2A6*1B were found to be of importance for the expression in both systems. Analysis of reporter enzyme degradation in HeLa cells showed that luciferase-3'-UTR-CYP2A6*1A had a half-life of approximately 4.9h as compared to 6.3h for luciferase-3'-UTR-CYP2A6*1B. In conclusion, we identified polymorphic motifs in the CYP2A6 3'-UTR of importance for CYP2A6 mRNA stabilization and enzyme expression. Such polymorphism has been described to influence the in vivo rate of nicotine elimination and possibly the cigarette consumption and risk of smoking induced lung cancer. PMID- 16378602 TI - Fibroblast heterogeneity in collagenolytic response to colchicine. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are important in various physiological and pathological conditions, including those that involve homeostasis of collagen. Drug induced regulation of MMP-1, other MMPs and TIMPs is critical in treatment of various diseases, e.g. the use of the plant alkaloid, colchicine. One possible factor that might explain the failure in colchicine-treatment of some patients is interindividual variability on the cellular level. To investigate the possible individual heterogeneity in response to colchicine, we studied the effect of colchicine-induced synthesis of collagenase from 32 different human skin fibroblast strains derived from both healthy individuals as well as individuals with different skin diseases. We showed that colchicine induced an increased synthesis of collagenase in 22 of 32 cases. This heterogeneity occurred in fibroblasts from healthy as well as diseased individuals. To determine if colchicine also affected the fibroblast synthesis of gelatinase, stromelysin and tissue inhibitors of MMPs, we investigated several individuals from a single family. The results showed that both colchicine responsive and non-responsive fibroblasts with respect to collagenase synthesis responded to colchicine by an increased stromelysin synthesis, while the synthesis of gelatinase and TIMP-1 were unaffected. As a whole, our results indicate that individual heterogeneity in collagenase response to colchicine treatment may partly explain some of the controversial results obtained with colchicine as a drug. PMID- 16378603 TI - Neuroprotective effects of a postischemic treatment with a bradykinin B2 receptor antagonist in a rat model of temporary focal cerebral ischemia. AB - Bradykinin, an endogenous nonapeptide produced by activation of the kallikrein kinin system, promotes neuronal tissue damage as well as disturbances in blood brain barrier function through activation of B2 receptors. In a rat model of focal cerebral ischemia, blockade of B2 receptors before initiation of ischemia with the B2 receptor antagonist, LF 16-0687 Ms, afforded substantial neuroprotection. In order to assess the potential clinical value of this approach, we evaluated the effect of LF 16-0687 Ms given at reperfusion following focal cerebral ischemia on local cerebral blood flow (LCBF), neurological outcome, and infarct size. Sprague-Dawley rats were subjected to MCA occlusion for 90 min by an intraluminal filament. Animals were assigned to one of four treatment arms (n = 7 each): (1) vehicle, (2) LF 16-0687 Ms (1.0 mg/kg/day), (3) LF 16-0687 Ms (3.0 mg/kg/day), or (4) LF 16-0687 Ms (10.0 mg/kg/day) given at reperfusion and repetitively over 2 days. Neurological recovery was examined daily, and infarct volume was assessed histologically on day 7 after ischemia. Physiological parameters and local CBF were not influenced by the treatment. Significant improvement of neurological outcome was observed on postischemic day 3 in animals receiving 1.0 and 3.0 mg/kg/day of LF 16-0687 Ms (P < 0.05). Inhibition of B2 receptors significantly reduced infarct volume in all treated animals predominantly in the cortex. B2 receptor blockade with LF 16-0687 Ms showed neuroprotective effectiveness even when therapy was initiated upon reperfusion, i.e. 90 min after induction of ischemia. Therefore, blockade of B2 receptors seems to be a promising therapeutic approach after focal cerebral ischemia, which deserves further experimental and clinical evaluation. PMID- 16378604 TI - A comparison of three techniques for quantitative carbohydrate analysis used in characterization of therapeutic antibodies. AB - A comparison of three techniques for quantitative analysis of galactosylation present on immunoglobulins is described. ESIMS, MALDI-TOF MS, and anion-exchange chromatography with fluorescence detection were evaluated in terms of repeatability, limit of quantitation, selectivity, and linearity. A recombinant monoclonal IgG was enzymatically modified in vitro to produce essentially completely galactosylated and degalactosylated forms of the immunoglobulin. Samples of known galactosylation levels were prepared by mixing the modified forms with the native form of the immunoglobulin. Good repeatability and linearity were demonstrated for all three assays (RSDs<1.0%, correlation coefficients>0.99). Differences in selectivity, sensitivity, and other performance aspects of the three techniques are also discussed in this paper. PMID- 16378605 TI - 9alpha,11beta-PGF2 and its stereoisomer PGF2alpha are novel agonists of the chemoattractant receptor, CRTH2. AB - CRTH2 is a recently described chemoattractant receptor for the prostaglandin, PGD(2), expressed by Th2 cells, eosinophils and basophils, and believed to play a role in allergic inflammation. Here we describe the potency of several PGD(2) metabolites at the receptor to induce cell migration and activation. We report for the first time that the PGD(2) metabolite, 9alpha,11beta-PGF(2), and its stereoisomer, PGF(2alpha), are CRTH2 agonists. 9alpha,11beta-PGF(2) is a major metabolite produced in vivo following allergen challenge, whilst PGF(2alpha) is generated independently of PGD synthetase, with implications for CRTH2 signalling in the presence or absence of PGD(2) production. PMID- 16378606 TI - Phytochromes from Agrobacterium tumefaciens: difference spectroscopy with extracts of wild type and knockout mutants. AB - Phytochromes are photoreceptors that occur in plants, fungi and bacteria, among others in the phytopathogen Agrobacterium tumefaciens. We constructed single and double knockout mutants of the two A. tumefaciens phytochromes Agp1 and Agp2. In liquid culture, the double mutant revealed a reduced growth rate, whereas the growth rates of the single mutants did not differ significantly from that of the wild type. Using these mutants, we analyzed the spectral properties of native A. tumefaciens phytochromes. A wild-type A. tumefaciens cell contains about 10 molecules of Agp1 and about 19 molecules of Agp2. Dark conversion of native Agp1 and Agp2 proceeds from Pfr to Pr and from Pr to Pfr, respectively, as has already been reported for the recombinant proteins. The spectral properties of recombinant and native Agp2 were significantly different. Mixing experiments with extracts from the double mutant and recombinant Agp2 imply that the spectral properties of Agp2 are modulated by components of the extract. PMID- 16378607 TI - Social suppression of female reproductive maturation and infanticidal behavior in cooperatively breeding Mongolian gerbils. AB - In several cooperatively breeding species, reproductively suppressed, nonbreeding females are attracted to infants and routinely provide alloparental care, while breeding females may attack or kill other females' infants. The mechanisms underlying the transition from alloparental to infanticidal behavior are unknown. In this study, we tested the hypothesis that this transition is associated with cessation of reproductive suppression and onset of ovarian activity in the Mongolian gerbil (Meriones unguiculatus), a cooperatively breeding rodent. Young female gerbils were housed with their natal family (FH), with a female pairmate (PH) or singly (SH). When females were either 11-13 or 16-18 weeks of age, we characterized their behavioral responses to an unfamiliar pup, reproductive development, and plasma progesterone, cortisol, and prolactin concentrations. In both age groups, FH females were significantly less likely to attack pups than PH or SH females and in fact never did so. FH females also had smaller ovaries and uteri, less developed scent glands, and lower progesterone levels, and were more likely to be anovulatory than PH or SH females, especially in the 11- to 13-week old age group. Prolactin concentrations did not differ with reproductive status but were significantly higher in females that did not attack pups than in those that did. We found no other significant associations, however, between reproductive or endocrine measures and behavioral responses to pups. These results suggest that cohabitation with the natal family inhibits both infanticidal behavior and reproductive maturation in young female gerbils but that these two effects may not be causally related. PMID- 16378608 TI - Comment on the article by Hegeman et al. regarding management of unstable distal radial fractures in women over 55. PMID- 16378609 TI - Salvage of failed dynamic hip screw fixation of intertrochanteric fractures. AB - Twenty-six patients with failed dynamic hip screw fixation of intertrochanteric fractures were included in this study. The mean age of the patients was 61 years (range, 38-84 years). Average limb shortening was 2.4 cm; 18 patients were treated with revision internal fixation and eight patients with prosthetic replacement. The decision depended on the physiological age of the patient, quality of bone, and condition of the femoral head and the acetabulum. The revision internal fixation group included DHS reinsertion in eight patients, valgus osteotomy and revision DHS fixation in six, while four patients were treated by valgus osteotomy and insertion of single-angled 130 degrees plate. The prosthetic replacement group included cemented Thompson endoprothesis in five patients and cemented total hip arthroplasty in three. The mean follow-up period was 31 months (range, 15-72 months). All patients of revision internal fixation group achieved fracture healing without bone grafting. Time to union averaged 17 weeks. Average gain in length was 2 cm Avascular necrosis of the femoral head occurred in one patient. Six patients of the prosthetic replacement group achieved good functional outcome and pain-free gait. The remaining two had unsatisfactory result. PMID- 16378610 TI - Previous cesarean scar exploration following vaginal delivery and hemorrhagic morbidity. PMID- 16378612 TI - Blood leukocyte count is a risk factor for intima-media thickening and subclinical carotid atherosclerosis in middle-aged men. AB - Leukocytosis is known to predict future cardiovascular events even in subjects without coronary heart disease (CHD), but its association with early atherosclerotic changes has remained less certain. The aim of the present study was to investigate how the blood leukocyte count compares with several other risk factors for CHD in determining carotid artery intima-media thickness (IMT) and subclinical carotid atherosclerosis in a population sample. Both carotid arteries were investigated with high-resolution B-mode ultrasound in a community-based sample of 219 randomly selected men aged 50-59 years to calculate the mean maximum IMT (MMax IMT) of 12 standard sites. Risk factor assessment included several traditional biochemical risk factors, blood pressure, maximal oxygen consumption and work load on ergometry, life-style habits and hematologic parameters. As genetic determinants, apolipoprotein E and A-IV polymorphisms were studied. According to multivariate regression analysis, age (P<0.0001), blood leukocyte count (P<0.0001) and systolic blood pressure (P<0.042) were the only significant predictors of MMax IMT. MMax IMT increased linearly from the lowest tertile of blood leukocyte count (1.14+/-0.20mm) to the second (1.18+/-0.25 mm) and to the highest tertile (1.25+/-0.27 mm, P=0.019). This difference remained significant after adjustment with age, systolic blood pressure and smoking (P=0.032). Leukocytes seem to have an independent role in the early arterial damage and they may reflect subclinical disease. This implies that leukocyte count is undervalued in the diagnostics and prognostics of carotid atherosclerosis. PMID- 16378611 TI - Decreased plasma CXCL16/SR-PSOX concentration is associated with coronary artery disease. AB - OBJECTIVES: To investigate for the first time whether the plasma CXCL16 concentration is altered in coronary artery disease (CAD) patients. BACKGROUND: Accumulating evidence suggests that the novel chemokine/scavenger receptor CXCL16/SR-PSOX is involved in the development of atherosclerosis and CAD. METHODS: Using ELISA we assessed the plasma CXCL16 concentration in 40 stable angina pectoris (SAP) patients, 17 unstable angina pectoris/non-ST-elevation myocardial infarction (UAP/non-STEMI) patients, 387 survivors of a first myocardial infarction (MI) and healthy control subjects (44 controls for SAP and UAP/non-STEMI patient groups and 387 controls for post-MI patients). RESULTS: SAP patients exhibited significantly lower median CXCL16 levels (2111 pg/ml) than the corresponding control subjects (2678 pg/ml) (P=0.0012). UAP/non-STEMI patients also appeared to have lower CXCL16 levels (2192 pg/ml) compared with controls (NS). Patients investigated 3 months after MI tended (P=0.07) to have lower CXCL16 levels (2529 pg/ml) than the corresponding controls (2638 pg/ml). There were no significant correlations between CXCL16 levels and different measures of CAD severity determined by quantitative coronary angiography in post-MI patients. Neither patients nor controls exhibited significant correlations between CXCL16 levels and plasma lipoprotein fractions, inflammatory cytokines, C-reactive protein or numbers of inflammatory cells in peripheral blood. CONCLUSIONS: The finding that lower plasma CXCL16 concentration is associated with CAD might indicate a potential atheroprotective function of CXCL16. PMID- 16378613 TI - Non-uniform strain distribution within rat cartilaginous growth plate under uniaxial compression. AB - Growth plates are highly inhomogeneous in morphology and composition. Mechanical loading can modulate longitudinal bone growth, though the mechanisms underlying this mechanobiology are poorly understood. The proximal tibial growth plates of six rats were tested in vitro under uniaxial compression to 5% strain, and confocal microscopy was used to track and capture images of fluorescently labeled cell nuclei with increasing applied strains. The local strain patterns through the growth plate thickness were quantified using texture correlation analysis. The technique of texture correlation analysis was first validated by comparing theoretical simulated strain maps generated from numerically distorted images. The texture correlation algorithm was sensitive to the grid size superimposed on the original image, but remained insensitive to parameters related to the size of the final image mask, which was searched by the correlation algorithm for each grid point of the original image. Within the growth plate, experimental strain distributions were non-uniform in all six specimens. Growth plates were mostly under compression strains. The strain distributions differed among the histomorphological zones of the growth plate, which was most obvious in specimens with regular growth plate shape: higher compressive strains (4-8 times higher than the applied 5% strain) were located mainly in regions overlapping the reserve and hypertrophic zones with lower compressive strains in the proliferative zone. This study documents the non-uniform mechanical behavior of growth plate across its three histological zones when exposed to compression. Further investigation is required to establish the significance of non-uniform strain fields during growth in vivo. PMID- 16378614 TI - Reversed phase ion-pair high performance liquid chromatographic gradient separation of related impurities in 2,4-disulfonic acid benzaldehyde di-sodium salt. AB - A reversed phase ion-pair gradient liquid chromatographic method has been developed and validated for purity determination of the hydrophilic compound 2,4 disulfonic acid benzaldehyde di-sodium salt (2,4-DSAD) containing both hydrophilic and more lipophilic related impurities. Mixtures of acetonitrile phosphate buffer containing tetrahexylammonium hydrogen sulfate as the ion pairing reagent were used as the mobile phase. A linear gradient, which generated simultaneous change in the concentration of organic modifier, buffer concentration and the concentration of ion-pairing reagent, was applied. The method allows detection of impurities at low levels (0.01% w/w). Excellent repeatability for both retention time (RSD< or =0.3%, n = 6) and detector response (RSD = 0.03%, n = 6 for the main peak and RSD = 6%, n = 6, for an impurity at 0.01 area% level) was obtained. The method was shown to be robust for routine analysis and has been successfully transferred to the quality control laboratory. PMID- 16378615 TI - Quantification of Fumaria officinalis isoquinoline alkaloids by nonaqueous capillary electrophoresis-electrospray ion trap mass spectrometry. AB - A capillary electrophoresis (CE) method using non-aqueous (NA) separation solutions combined with an ion trap mass spectrometer (MS and MS/MS) as detection device is presented for the separation, identification and quantification of isoquinoline alkaloids from Fumaria officinalis. The best results were obtained with a mixture of acetonitrile-methanol (9:1, v/v) containing 60mM ammonium acetate and 2.2M acetic acid as running electrolyte and an applied voltage of 30 kV. Electrospray MS measurements were performed in the positive ionization mode with isopropanol-water (1:1, v/v) as sheath liquid at a flow rate of 3 microl/min. Alkaloids were detected as [M+H](+)-ions and showed typical fragmentation patterns in MS/MS experiments. The developed assay was used for the quantification of seven isoquinoline alkaloids representing different structural subtypes in Fumariae herba extracts and F. herba containing phytopharmaceuticals. PMID- 16378616 TI - Development of an improved heated interface for coupling solid-phase microextraction to high-performance liquid chromatography. AB - The aim of the study described in this report has been the development and the evaluation of a new improved interface to be operated under continuous heating, for on-line coupling solid-phase microextraction (SPME) to high-performance liquid chromatography (HPLC). Heating is desirable to increase desorption rate and decrease carryover. The results obtained have been compared with that obtained by off-line desorption and online desorption without heating. The SPME HPLC interface described here has an inner volume of 60 microL, fixation for infinite points and a novel leak less sealing system. When the heating system was used, the area values were almost 10-fold higher than that obtained using the off line mode. The obtained chromatograms showed an increasing of the area and height of chromatographic peaks and proved the excellent performance and reproducibility of the interface developed in this work. PMID- 16378617 TI - Simultaneous determination of phenols (bibenzyl, phenanthrene, and fluorenone) in Dendrobium species by high-performance liquid chromatography with diode array detection. AB - A new method of analysis of 11 phenols, including five bibenzyls, three phenanthrenes, and three fluorenones, using high-performance liquid chromatography (HPLC)-diode array detection (DAD) was described. The separation of 11 phenols was effected by RP-HPLC (Beckman Coulter ODS column, 5 microm, 250 mm x 4.6 mm) using linear gradient elution systems of acetonitrile-1/1000 trifluoroacetic acid (TFA). Satisfactory separation of these compounds was obtained in less than 45 min. The method was validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). Good results were obtained with respect to repeatability (relative standard deviation (RSD)<3.5%) and recovery (85.77-104.92%). The developed method was applied to the simultaneous determination of 11 phenols from totally 31 Dendrobium species (mainly of medicinal plants) as well as other four samples from the similar genera as Pholidota, Flickingeria and Bulbophyllum. The range of the total amounts of bibenzyl, phenanthrene and fluorenone were found to from trace: 4.00, not detected (nd): 0.42 and nd: 0.24 microg mg(-1), respectively. PMID- 16378618 TI - Stainless steel electrospray probe: a dead end for phosphorylated organic compounds? AB - A study of the interaction of phosphorylated organic compounds with the stainless components of a liquid chromatography-electrospray ionisation-mass spectrometry system (LC-ESI-MS) was carried out to disclose a (forgotten?) likely pitfall in the LC-ESI-MS analysis of phosphorylated compounds. The retention behaviour of some representative compounds of different important classes of phosphorylated biomolecules such as nucleotides, oligonucleotides, phosphopeptides, phospholipids and phosphorylated sugars was investigated during their passage through the injector and the stainless steel electrospray capillary. It became clear that the stainless steel components within the LC-ESI-MS setup were able to retain and trap phosphorylated compounds when these compounds were introduced under acidic conditions (0.1% acetic acid). Their release from these stainless steel parts was accomplished by applying an extreme basic mobile phase (25-50% ammonium hydroxide, ca. pH 12). From the data collected one could conclude that the availability of a primary phosphate group appeared imperative but was not always sufficient to realise adsorption on a stainless surface. Furthermore, the number of phosphate moieties seemed to enhance the adsorption properties of the molecules and hence roughly correlated with the analyte fraction lost. Corrosion of the inner surface caused by the mobile phase and the electrospray process was found to be an important factor in the course of these adsorption phenomena. PMID- 16378619 TI - Evaluation of column bleed by using an ultraviolet and a charged aerosol detector coupled to a high-temperature liquid chromatographic system. AB - In this study, five different HPLC columns were heated to 200 degrees C using a homemade heating system which can be operated in temperature programmed mode. The column bleed as an indicator of induced degradation of the stationary phase material was evaluated using a charged aerosol detector (CAD) and an ultraviolet diode array detector (UV-DAD) at different wavelengths. The silica based C-18 stationary phase gave the highest bleed, and the carbon clad titanium dioxide column the lowest bleed. This was independent of both the detection technique and the wavelength. PMID- 16378620 TI - Flow splitting at the inlet electrode as a method for decreasing the electric current in electric field assisted liquid chromatography. AB - The combination of pressurized flow and electric field offers, with the use of capillary columns, several options for retention control. However, it has been shown that the utility of this technique is strongly limited by the high electric current that is generated at the high electric field strengths that are needed. We have earlier shown that the high current is a result of locally increased mobile phase ion concentration in the electric field, particularly around the inlet electrode. In this paper, we report that by splitting the mobile phase flow around the inlet electrode a relatively constant ion concentration around the electrode can be obtained and the high currents are there by reduced. PMID- 16378621 TI - Mixed micellization and interfacial properties of dodecyltrimethylammonium bromide and tetraethyleneglycol mono-n-dodecyl ether in absence and presence of sodium propionate. AB - Mixed micelle formation and interfacial properties of aqueous binary surfactant combinations of dodecyltrimethylammonium bromide (C12TAB) and tetraethyleneglycol mono-n-dodecyl ether (C12E4) at 30 degrees C in absence and presence of sodium propionate (NaPr) have been investigated. The critical micelle concentration, aggregation number, micropolarity and interfacial adsorption have been quantitatively estimated by surface tension and steady-state fluorescence measurements. The micellar and adsorption characteristics like composition, activity coefficients and mutual interaction parameters have been estimated following different theoretical treatments like that of Clint, Rubingh, Rodenas, Maeda, Blankschtein and Rosen. The analysis reveals very small mole fraction of cationic surfactant in both the mixed micelles and mixed monolayer inspite of synergism. Blankschtein's model predicts a continuous decrease in synergism due to the salt effect of NaPr; Rubingh's approach, on the contrary, indicates an increase in it above 30 mM of NaPr concentration. Aggregation number variation with NaPr indicates the same. Mixed monolayer shows better synergism compared to that in mixed micelles which increases with the addition of sodium propionate above 30 mM concentration. PMID- 16378622 TI - Temperature effect on the zeta potential and fluoride adsorption at the alpha Al2O3/aqueous solution interface. AB - The effect of temperature and pH on the zeta potential of alpha-Al2O3 and adsorption of fluoride ions at the alpha-Al2O3/aqueous solution interface has been investigated through electrophoretic mobility measurements and adsorption studies, to delineate mechanisms involved in the removal of fluoride ions from water using alumina as adsorbent. When the temperature increases from 10 to 40 degrees C, the pH of the point of zero charge (pH(pzc)) shifts to smaller values, indicating proton desorption from the alumina surface. The pH(pzc) increases linearly with 1/T, which allowed estimation of the standard enthalpy change for the surface-deprotonation process. Fluoride ion adsorption follows a Langmuir type adsorption isotherm and is affected by the electric charge at the alpha Al2O3/aqueous solution interface and the surface density of hydroxyl groups. Such adsorption occurs through an exchange between fluoride ions and surface-hydroxyl groups and it depends on temperature, pH, and initial fluoride ion concentration. At 25 and 40 degrees C, maximum fluoride adsorption density takes place between pH 5 and 6. Increasing the temperature from 25 to 40 degrees C lowers the adsorption density of fluoride. PMID- 16378623 TI - High plasticity of the hepatitis B virus capsid revealed by conformational stress. AB - Hepatitis B virus (HBV) replicates through reverse transcription inside its icosahedral nucleocapsid. The internal genome status is signaled to the capsid surface, predicting regulated conformational changes in the capsid structure. To probe their nature and extent, we imposed local conformational stress on the outer surface of HBV capsid-like particles, and monitored its consequences by electron cryomicroscopy and image reconstruction. The capsid structure had an enormous flexibility and robustness as a whole, as well as within the subunits, whose spikes were able to rotate by as much as 40 degrees against the distal interdimer contact sites. The likely hinge for the swiveling movement was the conserved Gly111 residue at the inner surface of the capsid. The stress imposed from the outside also affected the internal capsid organization, implying a specific route for the flow of conformational information between capsid interior and exterior as required for signaling of the genome status. PMID- 16378624 TI - Energy cost of infection burden: an approach to understanding the dynamics of host-pathogen interactions. AB - A mathematical model of long-term immune defense against infection was used to estimate the energy involved in the principal processes of immune resistance during periods of health and infection. From these values, an optimal level of energy was determined for immune response depending on infection burden. The present findings suggest that weak but prevalent pathogens lead to latent or chronic infection, whereas more virulent but less prevalent pathogens result in acute infection. This energy-based approach offers insight into the mechanisms of immune system adaptation leading to the development of chronic infectious diseases and immune deficiencies. PMID- 16378625 TI - Upregulation of heme oxygenase-1 by Epigallocatechin-3-gallate via the phosphatidylinositol 3-kinase/Akt and ERK pathways. AB - Heme oxygenase-1 (HO-1) is a cytoprotective enzyme activated by various phytochemicals and we examined the ability of Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, to upregulate HO-1 expression in endothelial cells (ECs). We demonstrate that EGCG induces HO-1 expression in a concentration- and time-dependent manner. Furthermore, EGCG-mediated HO-1 induction was abrogated in the presence of actinomycin D and cycloheximide, indicating that this upregulation of HO-1 occurred at the transcriptional level. EGCG also upregulates Nrf2 levels in nuclear extracts and increases ARE-luciferase activity. Furthermore, EGCG is the most potent inducer of HO-1 expression of the different green tea constituents that we analyzed, but had no detectable cytotoxic effects over the 25-100 microM dosage range. The inhibition of intracellular ROS production by N-acetylcysteine (NAC), glutathione (GSH), superoxide dismutase (SOD), catalase and the mitochondrial complex I inhibitor, rotenone, results in a decrease in EGCG-dependent HO-1 expression. In addition, we determined that tyrosine kinase is involved in EGCG induction of HO-1 as this is abrogated by genistein. ECs treated with EGCG exhibit activation of Akt and ERK1/2. In addition, pharmacological inhibitors of phosphatidylinositol 3-kinase and MEK1/2, which are upstream of Akt and ERK1/2, respectively, attenuate EGCG induced HO-1 expression. On the other hand, pretreatment of these cells with EGCG exerts significant cytoprotective effects against H2O2, suggesting that the induction of HO-1 is an important component in the protection against oxidative stress. Hence, EGCG is a novel phytochemical inducer of HO-1 expression and we further identify the principal underlying mechanisms involved in this process. PMID- 16378626 TI - Characterisation and differential expression of two very closely related G protein-coupled receptors, GPR139 and GPR142, in mouse tissue and during mouse development. AB - By searching the human and mouse genomic databases we found two G-protein-coupled receptors, GPR139 and GPR142, with characteristic motifs of the rhodopsin family of receptors. The gene for GPR139 maps to chromosome 7F1 of mouse and 16p12.3 of human and that for GPR142 to 11E2 of mouse and 17q25.1 of human. We isolated GPR139 from a cDNA library of adult mouse brain and GPR142 from a cDNA library of brains from 15-day-old mouse embryos. GPR139 mRNA was predominantly expressed in specific areas of human and mouse brains, whereas GPR142 mRNA showed a more ubiquitous expression both in the brain and in various peripheral glands and organs. A 50% identity and a 67% homology at the amino-acid level between the two receptors and only 20-25% identity with other G-protein-coupled receptors established them as a new subbranch within the phylogenetic tree and hints at a common or similar ligand(s). Preliminary results suggest that the cognate ligand is present in brain extracts and is, most likely, a small peptide. GPR139 signal transduction in Chinese hamster ovary cells requires coupling to an inhibitory G protein and is mediated by phospholipase C. Dimer formation may be necessary for proper function. PMID- 16378627 TI - Is interhemispheric communication disturbed when the two hemispheres perform on separate tasks? AB - The coordination of sensorimotor tasks involving both visual half-fields requires exchange of information between the brain hemispheres. So far, this interhemispheric information transfer has never been investigated under conditions where the two hemispheres receive different visual inputs and each hemisphere performs on a different task. The present study asked whether such conditions affect the transfer of information between the hemispheres, and if so, at which processing stage. We addressed these issues by pairing a face comparison task with a visual discrimination task (I-task) designed to interfere with the interhemispheric information transfer required for face comparison. One version of the I-task (experiment 1) required discrimination of the faces of John Lennon and Yoko Ono; the other version (experiment 2) required discrimination between the names 'JOHN' and 'YOKO'. Thus, the two I-task versions overlapped at early visual processing stages where visual feature analysis is carried out, but differed at later processing stages where words or faces are represented as objects. We found that both I-task versions disrupted the interhemispheric information transfer for the face comparison task. This indicates that when both hemispheres are occupied by separate tasks, interhemispheric communication is less efficient. In addition, our results suggest that the hemispheres exchange sensory information already at a rather early visual processing stage. Hence, visual feature analysis in one hemisphere is probably informed about feature analysis in the other hemisphere and may also be modulated by it. PMID- 16378628 TI - Progress towards countrywide control of schistosomiasis and soil-transmitted helminthiasis in Uganda. AB - Schistosomiasis caused by infection with Schistosoma mansoni is a serious public health burden in 38 of the 56 districts of Uganda. This article reviews the initial experience of the national control programme. Launched in 2003, this started with a pilot phase with the main aim of utilizing the experience to formulate feasible and appropriate methods of drug delivery. Overall, 432,746 people were treated and coverage was 91.4% in schools and 64.7% in communities. The issues raised by independent evaluators included that most communities did not participate in the selection of community drug distributors (CDD) and that teachers and CDDs needed refresher training mainly on health education and the management of side effects. As a way forward, it is suggested that the Ministry of Health should integrate deworming into the existing health infrastructure so that every time a child is reached for any health service, the child is also de wormed. PMID- 16378629 TI - The cowpox virus fusion regulator proteins SPI-3 and hemagglutinin interact in infected and uninfected cells. AB - The serpin SPI-3 and the hemagglutinin (HA) encoded by cowpox virus (CPV) block cell-cell fusion, and colocalize at the cell surface. wtCPV does not fuse cells, but inactivation of either gene leads to fusion. SPI-3 mAb added to wtCPV infected cells caused fusion, confirming that SPI-3 protein at the cell surface prevents fusion. The SPI-3 mAb epitope mapped to an 85-amino acid region at the C terminus. Removal of either 44 residues from the SPI-3 C-terminus or 48 residues following the N-terminal signal sequence resulted in fusion. Interaction between SPI-3 and HA proteins in infected cells was shown by coimmunoprecipitation. SPI 3/HA was not associated with the A27L "fusion" protein. SPI-3 and HA were able to associate in uninfected cells in the absence of other viral proteins. The HA binding domain in SPI-3 resided in the C-terminal 229 residues, and did not include helix D, which mediates cofactor interaction in many other serpins. PMID- 16378630 TI - Hantaan/Andes virus DNA vaccine elicits a broadly cross-reactive neutralizing antibody response in nonhuman primates. AB - At least four hantavirus species cause disease with prominent renal involvement hemorrhagic fever with renal syndrome (HFRS); and several hantavirus strains cause disease with significant pulmonary involvement-hantavirus pulmonary syndrome (HPS). The most prevalent and lethal hantaviruses associated with HFRS and HPS are Hantaan virus (HTNV) and Andes virus (ANDV), respectively. Here, we constructed a DNA vaccine plasmid (pWRG/HA-M) that contains both the HTNV and ANDV M gene segments. Rhesus macaques vaccinated with pWRG/HA-M produced antibodies that bound the M gene products (i.e., G1 and G2 glycoproteins), and neutralized both HTNV and ANDV. Neutralizing antibody titers elicited by the dual immunogen pWRG/HA-M, or single-immunogen plasmids expressing only the HTNV or ANDV glycoproteins, increased rapidly to high levels after a booster vaccination administered 1-2 years after the initial vaccination series. Memory responses elicited by this long-range boost exhibited an increased breadth of cross neutralizing activity relative to the primary response. This is the first time that hantavirus M gene-based DNA vaccines have been shown to elicit a potent memory response, and to elicit antibody responses that neutralize viruses that cause both HFRS and HPS. PMID- 16378631 TI - Variation in the ability of human influenza A viruses to induce and inhibit the IFN-beta pathway. AB - We investigated the ability of a selection of human influenza A viruses, including recent clinical isolates, to induce IFN-beta production in cultured cell lines. In contrast to the well-characterized laboratory strain A/PR/8/34, several, but not all, recent isolates of H3N2 viruses resulted in moderate IFN beta stimulation. Through the generation of recombinant viruses, we were able to show that this is not due to a loss of the ability of the NS1 genes to suppress IFN-beta induction; indeed, the NS1 genes behaved similarly with respect to their abilities to block dsRNA signaling. Interestingly, replication of A/Sydney/5/97 virus was less susceptible to pre-treatment with IFN-alpha than the other viruses. In contrast to the universal effect on dsRNA signaling, we noted differences in the effect of NS1 proteins on expression of interferon stimulated genes and also genes induced by a distinct pathway. The majority of NS1 proteins blocked expression from both IFN-dependent and TNF-dependent promoters by an apparent post-transcriptional mechanism. The NS1 gene of A/PR/8/34 NS1 did not confer these blocks. We noted striking differences in the cellular localization of different influenza A virus NS1 proteins during infection, which might explain differences in biological activity. PMID- 16378632 TI - c-Myc substitutes for Notch1-CBF1 functions in cooperative transformation with papillomavirus oncogenes. AB - Infection by high-risk human papillomaviruses (HPV) and persistent expression of the viral oncogenes E6 and E7 are causally linked to the development of cervical cancers. These oncogenes require additional events for the complete transformation of human epithelial cells. Although exaggerated levels of c-Myc are observed in many cases of cervical cancer, the actual function of c-Myc in the process of HPV-mediated transformation is unclear. Here, we show that analogous to activated alleles of Notch1, c-Myc can cooperate with E6/E7 in epithelial transformation and can substitute for CBF1-dependent signals generated by Notch1. In addition, dominant-negative forms of c-Myc block transformation by activated Notch1, E6 and E7, suggesting that c-Myc is required for HPV16-mediated transformation. PMID- 16378633 TI - Neutralization and infectivity characteristics of envelope glycoproteins from human immunodeficiency virus type 1 infected donors whose sera exhibit broadly cross-reactive neutralizing activity. AB - In this study, we tested the hypothesis that donors with broadly cross-reactive HIV-1 neutralizing (BCN) sera are infected with viruses encoding envelope glycoproteins (Envs) with unusual immunogenic properties. Cloned env genes were from samples of donors previously identified as having BCN antibodies (BCN donors) and from other donors not known to have such antibodies (non-BCN donors). Neutralization properties of viruses pseudotyped with BCN and non-BCN Envs were determined using BCN, non-BCN sera and broadly cross-neutralizing monoclonal antibodies (Mabs). BCN sera neutralized with higher frequency and geometric mean titers than non-BCN sera. Viruses pseudotyped with BCN Envs were mostly resistant to neutralization by anti-gp120 Mabs but tended to be more sensitive to the anti gp41 Mabs, 2F5 and 4E10 than non-BCN Env-pseudotyped viruses. Sequence analysis of clones obtained from sequential samples of two BCN donors revealed respective 2F5 epitope mutations T662A and K665T. The K665T mutation evolved as the predominant genotype in the respective donor, consistent with an escape mutation event. The A662T mutation reduced sensitivity to 4E10, as well as 2F5 and homologous sera, consistent with neutralization escape mutation and targeting of the 2F5 epitope region by the serum. Our study suggests that viruses infecting these BCN donors encoded Envs that may have been unusually competent for induction of antibodies against the membrane proximal epitope region (MPER) of gp41, and these Envs may be useful vaccine components. PMID- 16378634 TI - Hybrid photocatalysis/membrane treatment for surface waters containing low concentrations of natural organic matters. AB - Since the mid-1990s, numerous studies on the treatment of drinking water by photocatalysis have been reported. Once optimised, the photocatalytic process can completely degrade numerous natural and artificial organic compounds. In this study, a hybrid photocatalysis/membrane process was used as a polishing treatment of surface water containing a small concentration of natural organic matters (i.e. total organic carbon (TOC) concentration of around 3mg/L) which may be difficult to remove using conventional filtration or coagulation. An optimum pH of 4.5 and a TiO(2) concentration of 0.1g/L were found to lead to the highest removal efficiencies. The relative effect of the individual processes featuring in the hybrid system (UV radiation, TiO(2) adsorption and membrane filtration) was also assessed for different pH values. The membrane separation process was accounted to remove around 18% of the initial TOC concentration, while TiO(2) adsorption alone was generally responsible for less than 5% of TOC removal during the 120 min of the experiments. However, when the natural water was only radiated by UV light, up to 70% of TOC was removed. A synergetic effect was observed when the three processes (TiO(2), UV and membrane) were used together. Comparison of removal efficiencies obtained during real and model (International Humic Substance Society) waters treatment by photocatalysis is also presented, revealing the importance of the nature of the feed in this type of treatment. PMID- 16378635 TI - T-kininogen induces endothelial cell proliferation. AB - Basal proliferation of endothelial cells increases with age, and this might play a role in the etiology of age-related vascular diseases, as well as angiogenesis. Serum kininogen levels increase during aging in rats and humans, and T-kininogen (T-KG) can affect proliferative homeostasis in several cell models. Both kinins and kininogens have been shown previously to be angiogenic through activation of endothelial cell proliferation, and here we show that exposure of endothelial cells to T-KG results in vigorous cell proliferation, accompanied by ERK/AKT activation. In our experiments, the proliferative response requires B1 and B2 kinin receptors, even though kinins are not released from the precursor. We hypothesize that the age-related increase in T-KG could play a significant role in the age-related dysregulation of vascular physiology and function. PMID- 16378636 TI - Formulation and characterization of a novel fluoride-releasing dental composite. AB - OBJECTIVES: The aims of this study were to formulate a novel fluoride-releasing dental composite and to evaluate its mechanical properties, fluoride release and recharge capabilities, water sorption and solubility. METHODS: A fluoride releasing dimethacrylate monomer containing a ternary zirconium fluoride chelate was synthesized. Three experimental fluoride-releasing composites were fabricated with different monomer formulas (wt.%)-(1) Control A: 40 BisGMA/40 TEDMA/20 UEDMA; (2) EXPERIMENTAL: 20 F-releasing monomer/20 BisGMA/40 TEDMA/20 UEDMA; (3) Control B: the same formula as Control A except that it contained 10 wt.% (of total monomer) tetrabutylammonium fluoride (TBAF), which had a fluoride content equivalent to EXPERIMENTAL. All three materials had the same filler content: 55 wt.% silanized fluoroaluminosilicate particles (0.8 microm) and 10 wt.% silanized fumed silica (14 nm). All materials contained 0.17 wt.% camphorquinone (CQ) as a photoinitiator and 0.69 wt.% ethyl-4-dimethylaminobenzoate (4EDMAB) as an accelerator. The materials were tested for fluoride release (for 184 days), fluoride recharge, compressive and flexure strength, water sorption and solubility. The data were analyzed using ANOVA and Tukey-HSD tests, and Kruskal Wallis test. RESULTS: The experimental composite had significantly higher fluoride release and fluoride recharge capabilities than both Control composites. It had significantly better physical and mechanical properties than Control B. SIGNIFICANCE: The combined use of the fluoride-releasing dimethacrylate monomer and fluoride-releasing filler can provide sustained high fluoride release and recharge as well as acceptable mechanical and physical properties. Simply adding organic fluoride salt in the monomer yields composites with poor mechanical and physical properties. PMID- 16378637 TI - Color interaction of dental materials: blending effect of layered composites. AB - OBJECTIVES: To evaluate the in vitro blending effect (BE) of layered resin composites related to material, shade, and differences in color and translucency between compared materials. METHODS: Specimens made of two composites (2CS, n=5) consisted of the outer composite with an outer diameter of 10mm, 4mm thick and an inner diameter of 4mm, 2mm thick for the inner composite. Thus, the inner composite was encircled by a 3mm outer composite around its circumference and backed by a 2mm thick outer composite, to simulate a dental restoration surrounded by hard dental tissues. The outer composite was Palfique Estelite (PE, C2 shade, standard shade), while the inner composites were PE A2, B2 and C2 shades and corresponding shades of Point 4 (P4), Tetric Ceram (TC) and Filtek A110 (FA) composites (batch shades). Single-composite, disk-shaped specimens (1CS) of all five shades (D=10mm, 2mm thick, n=5) were made as well. Visual color assessments were done by six observers using a lightbooth and 1 (mismatch) to 5 (perfect match) scale. The BE was calculated as a difference in scores between corresponding 2CS and 1CS. Z-scores and corresponding BE values (BE(Z)) were calculated. 1CS were also evaluated using a spectrophotometer. RESULTS: Blending effect ranged from -0.4 to 2.2, while BE(Z) ranged from -0.6 to 3.0. Mean visual scores for 1CS and 2CS were 1.8 (1.2) and 2.2 (1.3), respectively. BE increased with a decrease in color difference (r=0.41) and increase of translucency parameter (TP, r=0.77). High agreement was recorded among pairs of observers for both 1CS, r=0.95 (0.03) and 2CS, r=0.96 (0.02). SIGNIFICANCE: Blending effect is composite and shade dependent. Quantifying of blending potential of dental materials might provide useful clinical information for dental professionals. PMID- 16378638 TI - Micro-architecture of calcium phosphate granules and fibrin glue composites for bone tissue engineering. AB - Calcium phosphate ceramics are currently used as bone graft substitutes in various types of clinical applications. Fibrin glue is also used in surgery due to its haemostatic, chemotactic and mitogenic properties. By combining these two biomaterials, new composite scaffolds were prepared. In this study, we attempt to analyse whether thrombin concentration in the fibrin glue could influence the properties of the composite. The association between fibrin glue and calcium phosphate ceramic granules was characterized at the ultra structural level. Micro and macroporous biphasic calcium phosphate ceramic granules with a diameter of 1 2mm composed of hydroxyapatite and beta-tricalcium phosphate (60/40) were associated to fibrin glue. The composites were observed by scanning and transmission electron microscopy and microcomputed tomography. Fibre thickness, porosity and homogeneity of the fibrin clot were modified by increased the thrombin concentration. Mixing fibrin glue with calcium phosphate granules (1:2) did not modify the microstructure of the fibrin clot in the composite. Nevertheless, thrombin interacted with the bioceramic by inducing the nucleation of crystalline precipitate at the ceramic/fibrin glue interface. Combining fibrin sealant and calcium phosphate ceramics could lead to new scaffolds for bone tissue engineering with the synergy of the properties of the two biomaterials. PMID- 16378639 TI - Wavefront-guided versus standard LASIK enhancement for residual refractive errors. AB - OBJECTIVE: To assess efficacy, safety, predictability, stability, and changes in higher-order aberrations (HOAs) and contrast sensitivity (CS) after wavefront guided and standard LASIK enhancement for the correction of residual refractive errors. DESIGN: Prospective, randomized, comparative clinical study. PARTICIPANTS: Twenty eyes of 20 consecutive patients (spherical equivalent [SE], 2.01+/-1.36 diopters [D]) treated with wavefront-guided Zyoptix Ablation Refinement software (ZAR) LASIK and 20 eyes of 20 consecutive patients (SE, 1.81+/-1.21 D) treated with standard Planoscan LASIK, both for residual refractive error enhancement. MAIN OUTCOME MEASURES: Efficacy, safety, predictability, stability, HOAs, and CS were evaluated before and after enhancement at 6 months' follow-up. METHODS: Uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA), manifest refraction, CS by means of the Functional Acuity Contrast Test, and HOAs by means of Zywave aberrometry were evaluated preoperatively and 6 months after retreatment. RESULTS: At 6 months postoperatively, UCVA was 20/25 or better in 100% of the eyes. Efficacy indexes were 1.09 for ZAR patients and 0.95 for Planoscan patients. No eyes lost > or =1 line of BCVA; in the ZAR group, 2 eyes gained 1 line and 6 eyes gained > or =2 lines; in the Planoscan group, 3 eyes gained 1 line. The ZAR group showed a percentage of eyes (94.4%) within the 0.5-D range in SE higher than that shown by the Planoscan group (88.8%). After 6 months, the HOA root mean square (RMS) increased on average by a factor of 1.44 for the Planoscan group (P = 0.003). No change or reduction in HOA RMS was found in the ZAR group (factor of 0.96; P>0.01). Contrast sensitivity was reduced in the Planoscan group only at the highest spatial frequency (18 cycles per degree; P<0.01). There was a significant reduction of CS as a function of HOA increase for the Planoscan group (P<0.0001). No changes were observed for the ZAR group at any spatial frequency (1.5-18 cycles per degree; P>0.01). CONCLUSIONS: Wavefront-guided LASIK using the ZAR algorithm is an effective and safe procedure for treatment of residual refractive errors. Wavefront-guided LASIK does not increase HOAs and does not modify CS compared with preoperative values. Wavefront-guided LASIK seems to be better than standard LASIK for retreatments. PMID- 16378640 TI - Apolipoprotein E variants and cognition in healthy individuals: a critical opinion. AB - The epsilon4 allele of apolipoprotein E (ApoE) is a well-established risk factor for late onset Alzheimer's disease (AD). This knowledge has generated interest in the role of ApoE variants in normal cognition. Varying degrees of cognitive dysfunction have been described in non-demented individuals with one or two epsilon4 alleles leading to suggestions that the gene plays a role in normal cognition or helps calibrate the aging process. In this paper, these hypotheses are critically evaluated. It is argued that ApoE variants play no role in cognitive development. Given the differential neurocognitive sequelae of normal aging and AD, we also suggest that accelerated aging is unlikely to account for the pattern of deficits observed in non-demented epsilon4 allele carriers. We conclude that the neuropsychological dysfunction reported in non-demented epsilon4 carriers is most likely to be the result of incipient AD. PMID- 16378641 TI - Temporal information processing in ADHD: findings to date and new methods. AB - The ability to perceive and represent time is a fundamental but complex cognitive skill that allows us to perceive and organize sequences of events and actions, and to anticipate or predict when future events will occur. It is a multidimensional construct, and a variety of methods have been used to understand timing performance in ADHD samples, which makes it difficult to integrate findings across studies. While further replication is needed, growing evidence links ADHD to problems in several aspects of temporal information processing, including duration discrimination, duration reproduction, and finger tapping. Neuroimaging studies of ADHD have also implicated cerebellar, basal ganglia, and prefrontal regions of the brain, which are believed to subserve temporal information processing. This line of research implicates more basic cognitive mechanisms than previously linked with ADHD and challenges researchers to develop and utilize innovative, multidisciplinary, scientific methods to dissect the various components of temporal information processing. Recent advances in neuroimaging, such as magnetoencephalography in collaboration with structural magnetic resonance imaging, can discriminate temporal processing at the level of a millisecond. This approach can lay the groundwork to provide a more precise understanding of neural network activity during different aspects and stages of temporal information processing in ADHD. PMID- 16378642 TI - Comparative non-radioactive RT-PCR assay: an approach to study the neurosteroids biosynthetic pathway in humans. AB - Polymerase chain reaction (PCR) is a powerful tool for qualitative evaluation of nucleic acid expression. PCR has been widely applied to measure DNA and RNA messages expression. Neurosteroids synthesized in the nervous system are potent modulators of synaptic activity and have been implicated in several neuropsychiatric disorders. To examine the possibility of an altered expression of the neurosteroidogenic metabolic enzymes in neurological diseases (like Parkinson's disease, PD) we developed a comparative non-radioactive RT-PCR assay to detect the mRNA levels of the peripheral benzodiazepine receptor, the 5alpha reductase type 1 and 3alpha-hydroxysteroid-oxidoreductase type 1 and 2 in lymphocytes obtained from PD patients. The results were compared with that obtained from simultaneous quantification of progesterone, 5alpha dihydroprogesterone and 3alpha,5alpha-tetrahydroprogesterone in the plasma and cerebro-spinal fluid of the same individuals using a gas chromatography mass spectrometry (GC/MS) technique. We found a significant decrease of the rate limiting enzyme 5alpha-R1 along with a significant decrease in plasma and CSF of the 3alpha,5alpha-tetrahydroprogesterone and of the 5alpha-dihydroprogesterone. Comparative RT-PCR assay, along with complimentary techniques (i.e. GC/MS), has the sensitivity, selectivity and dynamic range to allow specific and reliable quantization of the enzymes involved in the neurosteroids pathway and represent a valuable tool to assess their expression in human neuropsychiatric conditions. PMID- 16378643 TI - Expression of apoptosis-related proteins in peritoneal, ovarian and colorectal endometriosis. AB - Endometriosis is defined as the presence of endometrial glands and stroma outside the uterus. Apoptosis, a physiological process by which multicellular organisms eliminate superfluous cells, is altered in tumor tissue. Here we studied the expression of the apoptosis-related proteins p53, bcl-2, bax, p21 and fas in proliferative (n=9) and secretory (n=9) endometrium, and in peritoneal (n=11), ovarian (n=20) and colorectal (n=20) endometriosis, by qualitative and semi quantitative immunohistochemical methods using the percentage of positive cells and HSCORE analysis. In endometrium, p53, p21 and fas expression was low, whereas bax and bcl-2 expression was elevated. Using HSCORE analysis, only bcl-2 expression varied during the menstrual cycle (48.9+/-34.2% in the proliferative phase, 11.5+/-24.7% in the secretory phase, p=0.01). Using HSCORE analysis, p53 expression was higher in ovarian endometriosis than in peritoneal (p<0.0001) and colorectal endometriosis (p=0.03). P21 expression was higher in ovarian endometriosis than in peritoneal (p=0.01) and colorectal endometriosis (p=0.01). Bcl-2 expression was lower in ovarian endometriosis than in peritoneal (p=0.0002) and colorectal endometriosis (p<0.0001). Fas expression was higher in peritoneal endometriosis than in ovarian (p=0.02) and colorectal endometriosis (p=0.008). In conclusion, these results confirm the involvement of apoptosis in the pathogenesis of endometriosis. Moreover, expression of apoptosis-related proteins varies according to the location of endometriosis suggesting the involvement of different apoptotic pathways. PMID- 16378644 TI - Inflammation and the apopto-phagocytic system. AB - Although under normal conditions many cells die daily mainly by apoptosis in human tissues, inflammation does not occur. The redundant function of a relatively large number of molecules are available to recognize changes occurring on the surface of apoptotic cells, to opsonize the dead cells and to engulf the apoptotic cells previously opsonized or not. Several components of the innate immune system are utilized in this process, mainly soluble factors which bind to the distinct molecular pattern of apoptotic cells. These cells, unlike necrotic ones, do not induce the expression of inflammatory cytokines in phagocytic cells, they can even inhibit such a response and engage an active signaling process to elicit a direct anti-inflammatory effect. The molecular details of these signaling processes have not been clarified yet. Both professional and "amateur" cells can engulf apoptotic cells and mediate an anti-inflammatory action. Disturbance of these processes have significant roles in development of autoimmune diseases and highly malignant tumors. PMID- 16378645 TI - Control of Trypanosoma cruzi infection and changes in T-cell populations induced by a therapeutic DNA vaccine in mice. AB - Previous work showed that immunotherapy with a DNA vaccine encoding Trypanosoma cruzi antigen TSA-1 reduced cardiac tissue damage and improved survival in mice when administered during the acute or chronic phases of T. cruzi infection. In the present study, we investigated changes in T-cell populations induced by DNA vaccine immunotherapy. ICR mice were infected with 500 T. cruzi blood trypomastigotes and treated during the acute or chronic phases with two 100 microg doses of DNA vaccine. Analysis of stained splenocytes by flow cytometry indicated that the therapeutic vaccine induced a rapid increase in the number of CD4+ and CD8+ T cells in both the acute and chronic phases. Also, there was a rapid increase in T. cruzi-specific IFNgamma-producing CD8+ T cells following treatment during the chronic phase. The effects of these changes on the control of infection required longer time periods to be detectable but resulted in a reduction in myocarditis and T. cruzi parasite burden in both phases of the infection, as assessed by histopathologic analysis and semi-quantitative PCR detection of T. cruzi in cardiac tissue. These results suggest that DNA vaccines that induce CD8+ T-cells activity and IFNgamma production, would be good candidates for effective therapeutic vaccination against T. cruzi infection. PMID- 16378646 TI - The role of supramolecular protein complexes and membrane potential in transmembrane signaling processes of lymphocytes. AB - The formation of protein patterns in lymphocyte plasma membranes is analyzed in the light of past and, also, very recent experiments. The analysis surveys the lateral organization of major histocompatibility complex glycoproteins, intercellular adhesion molecule-1, interleukin-2 and -15 receptors, Kv1.3 K+ ion channels and the T-cell receptor as well as their behavior under different conditions. These molecules form small- and large-scale clusters in the membrane of human lymphocytes. Many of the association motifs occur in other investigated cell types. The conclusions point toward a possible role for ion channel activities, membrane potential changes and alterations of the lateral organization of proteins in transmembrane signaling and cytotoxic interactions. In our outlook new factors that potentially affect membrane protein cluster formation and interactions are discussed. A role for MHC glycoproteins in concentrating membrane proteins and organizing protein patterns is suggested, and the possibility that the membrane potential may modulate protein conformation and, thereby, affect protein-protein interactions is pointed out. A well-defined role for the presence of ion channels in the immune synapse is offered, which could explain the significance of ion channel accumulation in the immune synapse together with the T-cell receptor. PMID- 16378647 TI - Vertically oriented internal auditory canal in an 8-year-old with hearing loss. AB - To report a unique orientation of the internal auditory canal and possible association with congenital hearing loss. Retrospective chart review of an 8-year old Hispanic male with a mixed hearing loss. Uniquely abnormal orientation of the internal auditory canal in a patient with apparently normal cochleovestibular structures and a mixed hearing loss. A vertically oriented IAC is a rarely described anatomical anomaly of the temporal bone that may have associations with congenital hearing loss. PMID- 16378648 TI - Perceptual-motor adaptations in a synchronization task: the joint effects of frequency and motion coherence manipulations. AB - Two experiments were conducted to examine the human ability to adapt to a perturbation in a synchronization task. Five experimental signal conditions were tested using random-dot kinematograms, representing four conditions with different coherence levels (100%, 50%, 30% and 10%) and one target-alone condition. Within one trial, increasing or decreasing the frequency of the sinusoidally moving signal dots abruptly in the midst of each trial provoked a perturbation. The first experiment was aimed to clarify the process of adaptation to the new frequency situation. The second experiment explored the role of visual feedback about the arm's position on the participants' ability to adapt after the perturbation had occurred. The results clearly demonstrated that the synchronization performance gradually declined in function of the increasing number of randomly moving noise dots. In the 50% coherence condition, the participants were not or only partially able to adjust their arm movements to the new frequency situation. In addition, the provision of enhanced visual feedback about the arm's failed to improve one's adaptive ability. In general, these findings provided evidence for the important role of perceptual constraints on perception-action coupling in this type of synchronization task. PMID- 16378649 TI - Atrial stunning as predictor of early relapse into atrial fibrillation after cardioversion. AB - Although the high rate of success after cardioversion, less than 50% of patients maintain sinus rhythm for the first year. In view for the high percentage of relapse into atrial fibrillation, it is interesting to analyze the relationship between atrial stunning after cardioversion and relapse into atrial fibrillation. Thus, we evaluated 101 patients with atrial fibrillation and successful cardioversion. Atrial mechanical function was assessed by measures of transmitral peak A wave velocity, determined before and weekly after cardioversion during 1 month. Fifty-five percent of patient relapse into atrial fibrillation during follow-up. No significant differences were found in clinical and echocardiographic variables between the group with and without relapse. However, the group of patients who relapsed into atrial fibrillation showed a lower peak A wave velocity immediately after cardioversion than patients who maintain in sinus rhythm at month (0.44+/-0.27 vs. 0.60+/-0.38 m/s p<0.01). Impaired atrial function improves during the first 14 days after cardioversion. PMID- 16378650 TI - Real-time reverse transcription PCR for the quantification of the mntH expression of Salmonella enterica as a function of growth phase and phagosome-like conditions. AB - This article presents an experimental design for measuring the mRNA expression in Salmonella enterica of the mntH gene in phagosome-mimicking conditions. The expression of mntH was quantified by real-time reverse transcription PCR for different S. enterica strains of porcine origin under different biological growth conditions which mimicked the environment inside the phagosome. The expression of mntH and the different control genes (16S rRNA, rpoD and gmk) varied according to the growth phase. For mntH a maximum in the expression was detected in the early exponential phase. To obtain an accurate quantification and reliable comparison of the mntH expression in different S. enterica strains under various biological conditions, the ratio mntH mRNA level to the normalization factor was determined. The latter is the geometric mean of the RNA level of three housekeeping genes 16S rRNA, rpoD and gmk calculated by the geNorm program. MntH was basally expressed in all tested S. enterica strains and induced by hydrogen peroxide (H(2)O(2)) or ethylenediaminetetraacetic acid (EDTA) in Brain Heart Infusion. Under the nutrient limiting conditions of Sauton medium, the basal mntH expression was higher than in BHI, whereas H(2)O(2) induced the expression 40 times. A similar induction was obtained for Salmonella in porcine peripheral blood monocytes (PBM). PMID- 16378651 TI - Analyzing genes using closing and replicating circles. AB - During the past two years, significant breakthroughs have been achieved in genetic analyses through the application of technologies based on analytical DNA circularization reactions. Padlock probes and molecular inversion probes have enabled parallel, high-throughput single nucleotide polymorphism (SNP) genotyping at increased scales, whereas, at the other end of the analysis spectrum, DNA molecules in individual cells have been genotyped, in situ, using padlock probes and rolling-circle amplification (RCA). This review describes the recent developments in the technologies that use specific DNA circularization, coupled to DNA amplification through PCR or rolling-circle amplification, and addresses the great potential of these tools. PMID- 16378652 TI - On-line implant reconstruction in HDR brachytherapy. AB - BACKGROUND AND PURPOSE: To evaluate the accuracy of on-line planning in an Integrated Brachytherapy Unit (IBU) using dedicated image distortion correction algorithms, correcting the geometric distortion and magnetic distortion separately, and to determine the effect of the reconstruction accuracy on clinical treatment plans in terms of deviations in treatment time and dose. PATIENTS AND METHODS: The reconstruction accuracy has been measured using 20 markers, positioned at well known locations in a QA phantom. Treatment plans of two phantoms representing clinical implant geometries, have been compared with reference plans to determine the effect of the reconstruction accuracy on the treatment plan. Before clinical introduction, treatment plans of three representative patients, based on on-line reconstruction, have been compared with reference plans. RESULTS: The average reconstruction error for 10 in. images reduces from -0.6 mm (range -2.6 to +1.0 mm) to -0.2 mm (range -1.2 to +0.6 mm) after image distortion correction and for 15 in. images from 0.8 mm (range -0.5 to +3.0 mm) to 0.0 mm (range -0.8 to +0.8 mm). The error in case of eccentric positioning of the phantom, i.e. 0.8 mm (range -1.0 to +3.3 mm), reduces to 0.1 mm (range -0.5 to +0.9 mm). Correction of the image distortions reduces the deviation in the calculated treatment time of maximally 2.7% to less than 0.8% in case of eccentrically positioned clinical phantoms. The deviation in the treatment time or reference dose in the plans based on on-line reconstruction with image distortion correction of the three patient examples is smaller than 0.3%. CONCLUSIONS: Accurate on-line implant reconstruction using the IBU localiser and dedicated correction algorithms separating the geometric distortion and the magnetic distortion is possible. The results fulfill the minimum requirements as imposed by the Netherlands Commission on Radiation Dosimetry (NCS) without limitations regarding the usable range of the field of view of the image intensifier. However, the C-arm angle is limited to those angles for which magnetic distortion corrections have been obtained. PMID- 16378653 TI - Neuropeptide-Y Y2-receptor agonist, PYY3-36 promotes non-rapid eye movement sleep in rat. AB - PYY3-36 is a major component of the gut-brain axis and peripheral administration has been reported to exert significant effects on feeding, brain function and is more selective for neuropeptide Y2 receptor. Therefore, we investigated the effects of nocturnal intraperitoneal administration of single doses of PYY3-36 (30 and 100 microg/kg i.p.) on food intake, water intake and the sleep-wake cycle in rats. Sleep recordings were carried out in male Sprague-Dawley rats implanted with cortical electroencephalogram (EEG) and neck electromyogram (EMG) electrodes. The EEG, EMG, food intake and water intake were assessed. The electrographic recordings obtained were scored visually as rapid eye movement (REM) sleep, non-REM (NREM) sleep and wakefulness. PYY3-36 administration 15 min prior to dark onset significantly (p<0.05) increased non-rapid eye movement (NREM) sleep and decreased wakefulness. Analysis of the dark-period at 4-h time intervals showed that nocturnal administration of PYY3-36 (30 and 100 microg/kg) significantly suppressed wakefulness and increased non-REM sleep during the first 4-h time interval. Time spent in wakefulness was significantly decreased after administration of PYY3-36 (30 and 100 microg/kg) when compared with administration of vehicle. In addition, PYY3-36 (30 and 100 microg/kg i.p.) induced an increase in the time spent in NREM sleep. The nocturnal intraperitoneal administration of the lower dose of PYY3-36 (30 microg/kg) also significantly decreased food intake [F (2,23)=4.90, p<0.05] but had no effect on water intake. These findings suggest that PYY3-36 may play an important role in the enhancement of NREM sleep and feeding behavior. PMID- 16378654 TI - Enhanced anti-fibrotic activity of plasmid DNA expressing small interference RNA for TGF-beta type II receptor for a mouse model of obstructive nephropathy by cationized gelatin prepared from different amine compounds. AB - The objective of this study is to increase the transfection efficiency of a plasmid DNA expressing small interference RNA (siRNA) for transforming growth factor-beta receptor (TGF-betaR) by various cationized gelatins of non-viral carrier and evaluate the anti-fibrotic effect with a mouse model of unilateral ureteral obstruction (UUO). Ethylenediamine, putrescine, spermidine or spermine was chemically introduced to the carboxyl groups of gelatin for the cationization. The plasmid DNA of TGF-betaR siRNA expression vector with or without complexation of each cationized gelatin was injected to the left kidney of mice via the ureter to prevent the progression of renal fibrosis of UUO mice. Irrespective of the type of cationized gelatin, the injection of plasmid DNA cationized gelatin complex significantly decreased the renal level of TGF-betaR over-expression and the collagen content of mice kidney, in marked contrast to free plasmid DNA injection. It is concluded that retrograde injection of TGF betaR siRNA expression vector plasmid DNA complexed with the cationized gelatin is available to suppress the progression of renal interstitial fibrosis. PMID- 16378655 TI - Effect of quercetin on altered vascular reactivity in aortas isolated from streptozotocin-induced diabetic rats. AB - The present work examined ex vivo the acute effect of quercetin on diabetic rat aortic ring reactivity in response to endothelium-dependent (acetylcholine, ACh) and endothelium-independent (sodium nitroprusside, SNP) relaxants, and to the alpha(1)-adrenergic agonist phenylephrine (PE). Responses were compared to those of aortic rings from age- and sex-matched euglycemic rats. Compared to euglycemic rat aortic rings, diabetic rings showed less relaxation in response to ACh and SNP, and greater contraction in response to PE. Pretreatment with quercetin (10microM, 20min) increased ACh-induced relaxation and decreased PE-induced contraction in diabetic, but did not affect euglycemic rat aortic ring responses. Following pretreatment with the nitric oxide synthase inhibitor Nomega-nitro-l arginine methyl ester (l-NAME, 10microM), quercetin reduced PE-induced contractions in both aortic ring types, although l-NAME attenuated the reduction in the diabetic rings. Quercetin did not alter SNP vasodilatory effects in either ring type compared to their respective controls. These findings indicate that quercetin acutely improved vascular responsiveness in blood vessels from diabetic rats, and that these effects were mediated, at least in part, by enhanced endothelial nitric oxide bioavailability. These effects of quercetin suggest the possible beneficial effects of quercetin in vivo in experimental diabetes and possibly in other cardiovascular diseases. PMID- 16378656 TI - The battle of the fold: chaperones take on prions. AB - Protein conformational diseases, such as Alzheimer's, Parkinson's and Huntington's, affect a large portion of our aging population. Cells have evolved mechanisms for rescuing and recycling misfolded proteins, but these systems are not perfect. Chaperones can rescue misfolded proteins by breaking up aggregates and assisting in the refolding process. Proteins that cannot be rescued by refolding can be delivered to the proteasome by chaperones to be recycled. One class of 'misfolded' proteins, prions, appears to evade detection by this machinery and persist in a misfolded state. In fact, it seems that the prions usurp the refolding machinery and actually employ chaperones to propagate the prion state. Recent data has begun to uncover the mechanism behind this unique relationship. PMID- 16378657 TI - Carboplatin-based chemotherapy in patients with advanced non-small cell lung cancer and a poor performance status. AB - BACKGROUND: Poor performance status patients with advanced non-small cell lung cancer (NSCLC) have frequently been excluded from clinical trials due to the perception that they would have excessive treatment-related toxicity and a limited life expectancy. METHODS: A retrospective review of two multicenter trials centered at the University of North Carolina of patients who were treated with platinum-based chemotherapy for advanced NSCLC was conducted. Patients were divided into two subgroups based on Karnofsky performance status (KPS). Patients with a KPS = 70 were considered to have poor performance status, while patients with a KPS > or =80 were considered to have good performance status. RESULTS: Of the 387 patients, 19% (n = 73) had a poor performance status. The response rate (complete and partial responses) was similar between the two sub-groups (26% versus 28%); however, there was a difference in survival (p = 0.0004, log-rank test) between the groups. The median survival and 1-year survival rate for the poor performance status patients was 4.9 months and 21%, while the good performance status patients had a median survival of 8.4 months and a 1-year survival rate of 31%. The rate of National Cancer Institute (NCI) Common Toxicity Criteria (CTC) toxicities was similar between the two groups (p = 0.33). The percentage of patients receiving <4 cycles of therapy in the poor and good performance status was 55 and 39%, respectively (p = 0.012). CONCLUSIONS: Patients with poor performance status treated with platinum based chemotherapy have a similar rate of toxicity compared to good performance status patients. Their overall survival was lower despite a similar response to chemotherapy. PMID- 16378659 TI - Percutaneous CT-guided fine needle aspiration for lung cancer smaller than 2 cm and revealed by ground-glass opacity at CT. AB - The purpose of this retrospective study was to evaluate the value of preoperative percutaneous CT-guided fine needle aspiration biopsy (CTNB) for peripheral lung cancers less than 2 cm in size, especially in cases showing of ground-glass opacities (GGO). From 1999 to 2002, 151 small lung cancers were resected in Tokyo Medical University Hospital. Among them, 96 patients (63.6%) in whom the lesions were located in the outer half of the lung field underwent CTNB in order to obtain a preoperative diagnosis. The factors influencing the diagnostic yield were analyzed. The overall diagnostic yield of CTNB was 64.6%: 48.5% for lesions smaller than 10 mm, 62.5% for those 11-15 mm, and 83.9% for those 16-20 mm, respectively. The diagnostic yield in GGO-dominant lesions (GGO ratio < 50%) and solid-dominant lesions (GGO ratio < 50%) were 51.2% and 75.6% (p = 0.018). In the GGO-dominant group, the diagnostic yields were 35.2% for lesions smaller than 10 mm, 50.0% for those 11-15 mm, and 80.0% for those 16-20 mm. In the solid-dominant group, diagnostic yield was 62.5% for cases smaller than 10 mm, 75% for 11-15 mm and 85.7% for 16-20 mm, respectively. Satisfactory diagnostic yield (>80%) was obtained by CTNB in cases larger than 15 mm. CTNB is a useful diagnostic modality for peripheral small lung cancers; however, for GGO-dominant lesions, the preoperative diagnostic yield is not significantly better than for solid-dominant lesions. PMID- 16378658 TI - Prolonged gemcitabine infusion in advanced non-small-cell lung cancer with stable disease after gemcitabine 30-min infusion. AB - BACKGROUND: Although 30-min gemcitabine infusion has become the standard administration, pre-clinical and clinical studies have suggested the possibility that an infusion rate of 10 mg/m(2) per minute may be more effective. The main objective of this study was to investigate whether the pursuance of gemcitabine, administered at a prolonged infusion rate, was able to convert stable disease to objective response after two or three cycle of standard administration. The secondary end-point was the evaluation of the new schedule toxicity. PATIENTS AND METHODS: Thirty-eight patients, with stage IIIA-B and IV NSCLC already treated by two or three cycles of 30-min gemcitabine infusion, alone or in combination with cisplatin, were enrolled: 26 patients (aged <70 years) were treated with cisplatin 80 mg/m(2) on day 1 plus gemcitabine 1200 mg/m(2) over 120 min on day 1 and 8 every three weeks and 12 patients (aged > or =70 years) were treated with gemcitabine alone 1200 mg/m(2) over 120 min on day 1 and 8 every three weeks, for two courses. Simon's two stage minimax design was applied to calculate the sample size. Assuming p(0) (low conversion rate) 5%, p(1) (target conversion rate of interest) 20%, alpha error 0.05, beta error 0.10 a total of 29 evaluable patients had to be accrued during stage 1. In case at least one objective response was observed, a further nine evaluable patients had to be enrolled into the study during stage 2. The regimen was considered promising if > or =4 objective responses out of 38 evaluable patients were observed. RESULTS: Thirty-eight patients were evaluable for response and in five patients (with stable disease after two courses of gemcitabine 30' infusion) a partial response was observed (conversion rate 13.1%, 95% confidence interval 4.4-28%). Toxicities were more frequently observed with cisplatin plus 120-min gemcitabine infusion: grade 3-4 neutropenia, thrombocytopenia and anaemia in 28, 22 and 16% of the courses, respectively. CONCLUSIONS: The prolongation of gemcitabine infusion time is able to convert stable disease to partial response in 13% of the cases. The haematological toxicity seems enhanced with cisplatin plus gemcitabine prolonged infusion. PMID- 16378660 TI - Life cycle assessment of active and passive groundwater remediation technologies. AB - Groundwater remediation technologies, such as pump-and-treat (PTS) and funnel-and gate systems (FGS), aim at reducing locally appearing contaminations. Therefore, these methodologies are basically evaluated with respect to their capability to yield local improvements of an environmental situation, commonly neglecting that their application is also associated with secondary impacts. Life cycle assessment (LCA) represents a widely accepted method of assessing the environmental aspects and potential impacts related to a product, process or service. This study presents the set-up of a LCA framework in order to compare the secondary impacts caused by two conceptually different technologies at the site of a former manufactured gas plant in the city of Karlsruhe, Germany. As a FGS is already operating at this site, a hypothetical PTS of the same functionality is adopted. During the LCA, the remediation systems are evaluated by focusing on the main technical elements and their significance with respect to resource depletion and potential adverse effects on ecological quality, as well as on human health. Seven impact categories are distinguished to address a broad spectrum of possible environmental loads. A main point of discussion is the reliability of technical assumptions and performance predictions for the future. It is obvious that a high uncertainty exists when estimating impact specific indicator values over operation times of decades. An uncertainty analysis is conducted to include the imprecision of the underlying emission and consumption data and a scenario analysis is utilised to contrast various possible technological variants. Though the results of the study are highly site-specific, a generalised relative evaluation of potential impacts and their main sources is the principle objective rather than a discussion of the calculated absolute impacts. A crucial finding that can be applied to any other site is the central role of steel, which particularly derogates the valuation of FGS due to the associated emissions that are harmful to human health. In view of that, environmental credits can be achieved by selecting a mineral-based wall instead of sheet piles for the funnel construction and by minimising the steel consumption for the gate construction. Granular activated carbon (GAC) is exclusively considered as the treatment material, both in-situ and on-site. Here it is identified as an additional main determinant of the relative assessment of the technologies since it is continuously consumed. PMID- 16378661 TI - The LDLR locus in Alzheimer's disease: a family-based study and meta-analysis of case-control data. AB - Genetic linkage studies suggest the presence of an Alzheimer's disease (AD) risk gene on chromosome 19, acting independently of apolipoprotein E (apoE), a known AD risk factor on 19q13. The low density lipoprotein receptor (LDLR) is an interesting candidate because it maps within the linked interval, and is intimately involved in cholesterol homeostasis and the function of apoE. We tested three previously reported single nucleotide polymorphisms (SNPs) within LDLR in a large sample of discordant sibships from multiplex AD families, and failed to find evidence for genetic association with disease risk. In addition, we performed meta-analyses for SNP rs5925 on published data from five independent case control samples, but did not detect any significant summary odds ratios. Based on our data, it seems unlikely that these genetic variants in LDLR make a significant contribution to AD risk in the general population. PMID- 16378662 TI - Synthesis, antibacterial and antifungal activity of some new thiazolylhydrazone derivatives containing 3-substituted cyclobutane ring. AB - A series of Schiff bases, combining 2,4-disubstituted thiazole and cyclobutane rings, and hydrazone moieties in the same molecule, was synthesized, characterized and evaluated for screening antibacterial and antifungal activities on microorganisms, respectively, on four bacteria and Candida tropicalis. The structures of original compounds were confirmed by analytical and spectroscopic (FT-IR, (1)H NMR and (13)C NMR) methods and elemental analysis. Both the antibacterial and antifungal activities and MIC values of compounds were reported. Among the tested compounds, the most effective compound providing a MIC value of 16 microg ml(-1) are 2 against C. tropicalis and Bacillus subtilis and 3 against B. subtilis only. PMID- 16378663 TI - The development and characterization of H5 influenza virus vaccines derived from a 2003 human isolate. AB - The pandemic threat posed by highly pathogenic H5N1 influenza A viruses has created an urgent need for vaccines to protect against H5 virus infection. Because pathogenic viruses grow poorly in chicken eggs and their virulence poses a biohazard to vaccine producers, avirulent viruses produced by reverse genetics have become the preferred basis for vaccine production. Here, we investigated two key characteristics of potential H5 vaccine candidates: the hemaggutinin (HA) cleavage site sequence and its modification to attenuate virulence and the choice of background virus to provide a high-growth rate. We produced recombinant (6:2 reassortant) viruses that possessed a series of modified avirulent-type HA and neuraminidase genes, both of which were derived from an H5N1 human isolate. The other genes of these recombinant viruses were derived from donor virus strains known to grow well in eggs: the human strain A/Puerto Rico/8/34 (PR8) or an avian strain. All of the recombinant viruses grew well in eggs, were avirulent in chicks, and protected animals against infection with a wild-type virus. However, one of the recombinant viruses with an avian virus background acquired a mutation in the HA cleavage site sequence that conferred virulence potential to this virus. Moreover, vaccine candidates with the avian virus background were more virulent than those with the human virus background. We conclude that 6:2 recombinant viruses with a PR8 background are more suitable than those with an avian virus background for vaccine development and that the HA cleavage site sequence must be modified to minimize the potential for a vaccine virus to convert to a virulent form. PMID- 16378664 TI - F4 (K88) fimbrial adhesin FaeG expressed in alfalfa reduces F4+ enterotoxigenic Escherichia coli excretion in weaned piglets. AB - Transgenic plants are attractive bioreactors to large-scale production of recombinant proteins because of their relatively low cost. This study reports for the first time the use of transgenic plants to reduce enterotoxigenic Escherichia coli (ETEC) excretion in its natural host species. The DNA sequence encoding the major subunit and adhesin FaeG of F4+ ETEC was transformed into edible alfalfa plants. Targeting of FaeG production to chloroplasts led to FaeG levels of up to 1% of the total soluble protein fraction of the transgenic alfalfa. Recombinant plant-produced FaeG (pFaeG) remained stable for 2 years when the plant material was dried and stored at room temperature. Intragastric immunization of piglets with pFaeG induced a weak F4-specific humoral response. Co-administration of pFaeG and the mucosal adjuvant cholera toxin (CT) enhanced the immune response against FaeG, reflected a better induction of an F4-specific immune response. In addition, the intragastric co-administration of CT with pFaeG significantly reduced F4+ E. coli excretion following F4+ ETEC challenge as compared with pigs that had received nontransgenic plant material. In conclusion, transgenic plants producing the FaeG subunit protein could be used for production and delivery of oral vaccines against F4+ ETEC infections. PMID- 16378665 TI - Immunologic enhancement of compound Chinese herbal medicinal ingredients and their efficacy comparison with compound Chinese herbal medicines. AB - Two compound Chinese herbal medicinal ingredients (cCIs) were prepared respectively with epimedium polysaccharide (EPS) plus propolis flavone (PF) and astragalus polysaccharide (APS) plus ginsenoside (GS). Also, two compound Chinese herbal medicines (cCMs) with the same ingredient content as corresponding cCIs were made with the extracts of epimedium plus propolis and astragalus plus ginseng. In rabbit immune trial, two cCIs, physiological saline in the control, were respectively injected to the rabbits vaccinated with inactivated rabbit hemorrhagic disease vaccine. On Days 3, 7, 10, 14, 21, 28, 35 and 42 after vaccination, the dynamic changes of serum antibody titers were determined by hemagglutination inhibition (HI) test. In chicken immune trial, all of cCIs and cCMs were mixed respectively with inactivated Newcastle disease vaccine virus to vaccinate chickens, taking oil-adjuvant and non-adjuvant vaccine as controls. On Days 7, 14, 21, 28, 35 and 42 after vaccination, the dynamic changes of peripheral lymphocyte proliferation and serum antibody titers were tested respectively by MTT method and HI test method. The results showed that both cCIs could significantly raise antibody titer in rabbits, which the effect of compound Chinese herbal medicinal ingredients 1 (cCIs 1) was better than that of compound Chinese herbal medicinal ingredients 2 (cCIs 2). All of cCIs and cCMs could markedly promote lymphocyte proliferation and enhance antibody titer in chickens, which was similar to oil adjuvant, the immunologic enhancement of cCIs were slightly superior to that of the cCMs. PMID- 16378666 TI - 137Cs in freshwater fish in Finland since 1986--a statistical analysis with multivariate linear regression models. AB - The accident at the Chernobyl nuclear power plant in 1986 significantly elevated the 137Cs levels of fish in Finnish lakes. About 6200 fish samples from 390 lakes comprising 20 species have been analysed for 137Cs since 1986. The sizes of the lakes varied from a few hectares to about 1000 km2. Activity concentrations of 137Cs in fish still varied widely in 2003, from 16 to 6400 Bq/kg fresh weight. This paper presents the results of statistical analyses with multivariate linear regression models carried out on the empirical data collected since 1986. The statistical analysis resulted in separate models for two time periods describing temporal changes of 137Cs in fish. The explanatory variables were fish species with various feeding habits, the size class of the lake, municipal division, drainage area, time since the deposition and deposition level of the municipality. The calculated values for 137Cs in fish did not differ statistically significantly from the observed values in the validation data. The explanatory variables explained 58% (the first time period) and 72% (the second time period) of the total variability of 137Cs in fish. PMID- 16378667 TI - Lower extremity biomechanics during the landing of a stop-jump task. AB - BACKGROUND: Literature shows that landing with great impact forces may be a risk factor for knee injuries. The purpose of this study was to examine the relationships among selected lower extremity kinematics and kinetics during the landing of a stop-jump task. METHODS: Landmark coordinates and ground reaction forces during a stop-jump task were collected. Lower extremity joint angles and resultants were reduced. Pearson correlation coefficients among selected lower extremity kinematics and kinetics were determined. FINDINGS: The hip flexion angular velocity at the initial foot contact had significant correlation with peak posterior and vertical ground reaction forces (r = -0.63, P < 0.001, r = 0.48, P < 0.001) during the landing of the stop-jump task. The knee flexion angular velocity at the initial foot contact also had significant correlation with peak posterior and vertical ground reaction force (r = -0.49, P < 0.001, r = -0.06, P < 0.001) during the landing of the stop-jump task. Peak proximal tibia anterior shear force and peak knee extension moment during landing of the stop jump task had significantly correlation with the corresponding posterior and vertical ground reaction forces (r > 0.51, P < 0.001). INTERPRETATION: A large hip and knee flexion angles at the initial foot contact with the ground do not necessarily reduce the impact forces during the landing of the stop-jump task, but active hip and knee flexion motions do. Hip joint motion at the initial foot contact with the ground appears to be an important technical factor that affects anterior cruciate ligament loading during the landing of the stop-jump task. PMID- 16378669 TI - Malignant catatonia with severe bronchorrhea and its response to electroconvulsive therapy. AB - A 21-year-old female presented excitement, auditory hallucination, monologue, and insomnia. After 1 week of risperidone administration, she showed hyperthermia, salivation, and muscle rigidity. Risperidone was discontinued, but stupor, convulsions, and respiratory distress developed. In the intensive care unit where she was transferred, catatonic symptoms such as stupor or excitement, catalepsy, and negativism were prominent. In addition, severe bronchorrhea causing respiratory failure was observed. Her catatonic symptoms, hyperthermia, and bronchorrhea resolved by ECT. After recovery, affective flattening, alogia, and avolition remained. The final diagnosis was MC associated with schizophrenia. This report suggests that MC may be complicated by severe bronchorrhea, but this condition responds to ECT. PMID- 16378668 TI - Spinal manipulation force and duration affect vertebral movement and neuromuscular responses. AB - BACKGROUND: Previous study in human subjects has documented biomechanical and neurophysiological responses to impulsive spinal manipulative thrusts, but very little is known about the neuromechanical effects of varying thrust force-time profiles. METHODS: Ten adolescent Merino sheep were anesthetized and posteroanterior mechanical thrusts were applied to the L3 spinous process using a computer-controlled, mechanical testing apparatus. Three variable pulse durations (10, 100, 200 ms, force = 80 N) and three variable force amplitudes (20, 40, 60 N, pulse duration = 100 ms) were examined for their effect on lumbar motion response (L3 displacement, L1, L2 acceleration) and normalized multifidus electromyographic response (L3, L4) using a repeated measures analysis of variance. FINDINGS: Increasing L3 posteroanterior force amplitude resulted in a fourfold linear increase in L3 posteroanterior vertebral displacement (p < 0.001) and adjacent segment (L1, L2) posteroanterior acceleration response (p < 0.001). L3 displacement was linearly correlated (p < 0.001) to the acceleration response over the 20-80 N force range (100 ms). At constant force, 10 ms thrusts resulted in nearly fivefold lower L3 displacements and significantly increased segmental (L2) acceleration responses compared to the 100 ms (19%, p = 0.005) and 200 ms (16%, p = 0.023) thrusts. Normalized electromyographic responses increased linearly with increasing force amplitude at higher amplitudes and were appreciably affected by mechanical excitation pulse duration. INTERPRETATION: Changes in the biomechanical and neuromuscular response of the ovine lumbar spine were observed in response to changes in the force-time characteristics of the spinal manipulative thrusts and may be an underlying mechanism in related clinical outcomes. PMID- 16378670 TI - The resuscitation greats. Cemil Topuzlu Pasha: one of the forgotten pioneers in the history of open chest cardiac massage. AB - One of the pioneers of open chest cardiac massage was Cemil Topuzlu Pasha, in Turkey. He presented his experience in open heart cardiac massage in several papers published in Ottoman and French and German. On 27 August 1903 one of his patients undergoing external urethrotomy under chloroform anaesthesia developed cardiac arrest and he performed open chest cardiac massage. He reported his approach to this event and discussed the literature regarding resuscitation available at the beginning of 20th century. Meanwhile he had defined "Do Not Resuscitate" code in cases involving serious heart disease and other diseases where life expectancy is very short. The Ottoman version of his report is translated in this paper. PMID- 16378671 TI - Warming of patients with accidental hypothermia using warm water pleural lavage. AB - In all, five patients with accidental hypothermia below 32 degrees C are described. All were unconscious and in mortal danger, but with an intact circulation. The youngest was 11 years and the oldest 85 years of age. The two oldest patients suffered from critical hypothermia only, while the other cases were complicated by other trauma and drug poisoning. All were warmed using pleural lavage with warm saline. All were discharged to their own homes neurologically intact. PMID- 16378672 TI - Post-shock myocardial stunning: a prospective randomised double-blind comparison of monophasic and biphasic waveforms. AB - INTRODUCTION: Compared with monophasic defibrillation, biphasic defibrillation is associated with less myocardial stunning and earlier activation of sodium channels. We therefore hypothesised that earlier sodium channel activation would result in earlier restoration of the first sinus beat following elective DC cardioversion. METHODS: Adults undergoing elective DC cardioversion were randomised to receive either monophasic or biphasic escalating transthoracic shocks. The ECG was recorded electronically during defibrillation and the time from delivery of the shock to restoration of the first sinus beat, measured from the beginning of the 'P' wave, was calculated. RESULTS: Seventy four patients were studied. Data were unavailable from 18 patients. There was no demographic difference between groups. Median time to the first sinus beat following monophasic defibrillation (n=25) was 3.66 s (95% CI 2.55-4.61 s) and following biphasic defibrillation (n=33) was 2.21s (95% CI 1.76-2.56 s; P 0.05). Nevertheless, elongation at break of films prepared at pH 3 was higher than that at pH 11 (P < 0.05). The acidic and alkaline conditions had no effect on water vapor permeability of the films obtained. However, the film prepared at acidic condition was more yellowish than that prepared at alkaline pH. Protein content influenced the mechanical properties and color of films. PMID- 16378727 TI - Facing the genetic heterogeneity in neuromuscular disorders: linkage analysis as an economic diagnostic approach towards the molecular diagnosis. AB - The identification of an ever increasing number of gene defects in patients with neuromuscular disorders has disclosed both marked phenotype and genotype variability and considerable disease overlap. In order to offer an economic strategy to characterise the molecular defect in patients with unclassified neuromuscular disorders, we designed DNA marker sets for linkage analysis of 62 distinct neuromuscular disorders gene loci, including all known muscular dystrophies, congenital myopathies, congenital myasthenic syndromes and myotonias. Genotyping of marker loci of 140 clinically well-characterised families with unclassified neuromuscular disorders reduced the number of candidates to one or two genes in 49 % of the families. Subsequent mutation analysis and genome-wide scans enabled the determination of the genetic defect in 31 % of the families including the identification of a new gene and a new mutation in an unexpected candidate gene. This highlights the effective application of this approach both for diagnostic strategies as well as for the identification of new loci and genes. PMID- 16378728 TI - A modelling study of a non-concerted hydrolytic cycloaddition reaction by the catalytic antibody H11. AB - H11 is the first antibody reported to have dual activity as a non-concerted, Diels-Alderase and hydrolytic catalyst. It was previously shown to catalyse the cycloaddition of acetoxybutadiene 1a to N-alkyl maleimides 2 to afford hydroxy substituted bicyclic adducts 3 with a 30% ee of a major isomer. To better understand this mechanism and the partial stereospecificity, a homology model of H11 was constructed and used in docking studies to evaluate potential antibody ligand complexes. The model suggested the hydrolytic nature of H11 was due to Glu 95H acting as a catalytic base, and evaluation of the shape complementarity of the proposed antibody-ligand complexes confirmed at a semi-quantitative level the observation that the major enantiomer is produced in a 30% ee. PMID- 16378729 TI - New leads of metallo-beta-lactamase inhibitors from structure-based pharmacophore design. AB - We have applied pharmacophore generation, database searching, docking methodologies, and experimental enzyme kinetics to discover new structures for design of di-zinc metallo-beta-lactamase inhibitors. Based on crystal structures of class B1 metallo-beta-lactamases with a succinic acid and a mercapto carboxylic acid inhibitor bound to the enzyme, two pharmacophore models were constructed. With the Catalyst program, these pharmacophores were used to search the ACD database, which provided a total of 74 hits representing four different chemical classes of compounds: Dicarboxylic acids, phosphonic and sulfonic acid derivatives, and mercapto-carboxylic acids. All hits were docked into different metallo-beta-lactamases (from classes B1 and B3) using the GOLD docking program. A selection scheme based on the GOLD scores, the Catalyst fit and shape values, and the size of the compounds (molecular weight, surface area, and number of rotatable bonds) was developed and thirteen compounds representing all four chemical classes were selected for experimental studies. Three compounds with new scaffolds hitherto not present in metallo-beta-lactamase inhibitors have IC50 values less than 15 microM and may serve as starting points in the design of metallo-beta-lactamase inhibitors. PMID- 16378730 TI - Synthesis and study of the cancer cell growth inhibitory properties of alpha-, gamma-tocopheryl and gamma-tocotrienyl 2-phenylselenyl succinates. AB - Vitamin E succinate selenium-conjugated molecules were synthesized and their apoptogenic properties were evaluated. 4-Methyl-2-phenylselenyl succinate (4) was prepared by the reaction of sodium benzeneselenolate with 2-bromosuccinic anhydrite in methanol solution. The methyl ester was converted to the acid (5) by hydrolysis with aqueous hydrochloric acid. Reaction of the 2-phenylselenyl succinic anhydrite (6) with alpha-tocopherol (1a), gamma-tocopherol (1c), and gamma-tocotrienol (2c) in acidic conditions gave the respective esters. The free radical scavenging properties of alpha-tocopheryl-2-phenylselenyl succinate (7), gamma-tocopheryl-2-phenylselenyl succinate (8), and gamma-tocotrienyl-2 phenylselenyl succinate (9) were evaluated in comparison with those of alpha tocopheryl succinate (10), gamma-tocopheryl succinate (11), and gamma-tocotrienyl succinate (12), respectively, and the free tocopherols and gamma-tocotrienol. Compounds 7-9 induced a statistically significant decrease in prostate cancer cell viability compared to 10-12, respectively, or 5, exhibiting features of apoptotic cell death and associated with caspase-3 activation. These data show that structural modifications of vitamin E components by 5 enhance their apoptogenic properties in cancer cells. PMID- 16378731 TI - Identification of nitrated proteins in Alzheimer's disease brain using a redox proteomics approach. AB - Nitric oxide (NO) has been implicated in the pathophysiology of a number of neurodegenerative diseases including Alzheimer's disease (AD). In the present study, using a proteomics approach, we identified enolase, glyceraldehyde-3 phosphate dehydrogenase, ATP synthase alpha chain, carbonic anhydrase-II, and voltage-dependent anion channel-protein as the targets of nitration in AD hippocampus, a region that shows a extensive deposition of amyloid beta-peptide, compared with the age-matched control brains. Immunoprecipitation and Western blotting techniques were used to validate the correct identification of these proteins. Our results are discussed in context of the role of oxidative stress as one of the important mechanisms of neurodegeneration in AD. PMID- 16378732 TI - Constitutive expression of IL-2Rbeta chain and its effects on IL-2-induced vascular leak syndrome. AB - IL-2-induced vascular leak syndrome (VLS) is an important mechanism explaining the toxic effects of this cytokine and limiting its therapeutic use. We previously characterized a mouse model of IL-2-induced pulmonary VLS used to demonstrate that NK lymphocytes are involved in early/acute phase VLS (after one IL-2 injection). We also showed that NK cells and polymorphonuclear neutrophils (PMN) are involved in the late/chronic phase of the syndrome (after four daily IL 2 injections). In this study we use our mouse model to evaluate the role played by the IL-2 receptor (IL-2R) in VLS induction. Mouse and human IL-2R are different since the mouse IL-2Rbeta chain does not recognize IL-2. Here, we compare the acute and late VLS responses in human IL-2Rbeta transgenic and C57BL/6 wild type mice. Parameters linked to early phase VLS (bronchoconstriction and PMN mobilization) are enhanced in human IL-2Rbeta transgenic mice. By contrast, parameters used to measure late events (protein leakage and edema) are similar in human IL-2Rbeta transgenic mice and C57BL/6 wild type animals. However, after four IL-2 injections, the cellular content of the bronchoalveolar lavage fluids was different between the two types of animals. This study also characterizes a humanized animal model that could be further used to study human IL-2 activity and side effects in vivo. PMID- 16378733 TI - Regulatory cells and human cancer. AB - While there is now little debate about the existence or relevance of regulatory populations of T cells to a variety of human diseases, including cancer, there is considerable debate about the ontogeny, phenotype, and mechanisms of action of given regulatory T cell populations. This review will limit itself to discussion to two distinct populations of CD4+ regulatory T cells: T cells co-expressing the CD25 receptor (Tregs), and type 1 regulatory (Tr1) T cells. Attention will be focused on the definition and role of these regulatory T cell populations in human cancers. PMID- 16378734 TI - Purification, characterization, and mass spectrometric sequencing of a medium chain acyl-CoA synthetase from mouse liver mitochondria and comparisons with the homologues of rat and bovine. AB - Medium chain acyl-CoA synthetases catalyze the first reaction of amino acid conjugation of many xenobiotic carboxylic acids and fatty acid metabolism. This paper reports studies on purification, characterization, and the partial amino acid sequence of mouse liver enzyme. The medium chain acyl-CoA synthetase was isolated from mouse liver mitochondria. The purified enzyme catalyzes this reaction not only for straight medium chain fatty acids but also for aromatic and arylacetic acids. Maximal activity was found with hexanoic acid. High activities were obtained with benzoic acid having methyl, pentyl, and methoxy groups in the para- or meta-positions of the benzene ring. However, the enzyme was less active with valproic acid and ketoprofen. Salicylic acid exhibited no activity. The medium chain acyl-CoA synthetases from mouse and bovine liver mitochondria were subjected to in-gel tryptic digestion, followed by LC-MS/MS sequence analysis. The amino acid sequence of each tryptic peptide of mouse liver mitochondrial medium chain acyl-CoA synthetase differed from that from bovine liver mitochondria only in one or two amino acids. LC-MS/MS analysis provided the information about these differences in amino acid sequences. In addition, we compared the properties of this protein with the homologues from rat and bovine. PMID- 16378735 TI - Effect of age on markers for monoaminergic neurons of normal and MPTP-lesioned rhesus monkeys: a multi-tracer PET study. AB - The binding of three tracers for monoaminergic terminals was mapped in the brain of healthy young (N=6) and healthy old rhesus monkeys (N=4), aged monkeys with mild unilateral intracarotid MPTP lesions (N=3), and monkeys of intermediate age with severe systemic MPTP lesions (N=6). The ligand for monoaminergic vesicles (+)-[(11)C]dihydrotetrabenazine (+DTBZ) had a mean binding potential (pB) of 1.4 in striatum of the healthy young monkeys, which was reduced by 20% in putamen of the old monkeys. The catecholamine transporter ligand (+)-[(11)C]methylphenidate (+MP) had a mean pB of 1.3 in striatum of the young monkeys, which was reduced by 40% in caudate and putamen of the old monkeys. The DOPA decarboxylase substrate [(18)F]fluoro-l-DOPA (FDOPA) had a mean decarboxylation coefficient (k(3)(S)) of 0.4 h(-1) in striatum of the young group, and was not significantly reduced in the aged group. Of the three ligands, only +DTBZ pB was significantly reduced in striatum of the small group of animals with mild unilateral lesions. In the group with systemic MPTP lesions, the mean reduction of the binding of the three ligands was 80% in the caudate and putamen. However, the decline in +MP pB in the ventral striatum (-75%) exceeded the declines of +DTBZ pB and FDOPA k(3)(S) in that region (-65%), suggesting that compensatory down-modulation of uptake sites may occur in the striatal regions with the least dopamine depletion. Binding of all three ligands was reduced by 50% in the anterior cingulate cortex and in the thalamus, suggesting toxicity of MPTP for extrastriatal catecholamine innervations. +DTBZ binding in the hypothalamus, presumably mainly in serotonin fibers, was unaffected by systemic MPTP treatment. Of the three tracers, +DTBZ was most sensitive for detecting MPTP-induced dopamine depletion in monkey striatum. PMID- 16378736 TI - Cytotoxicity, haematotoxicity and genotoxicity of high molecular mass arborescent polyoxyethylene polymers with polyglycidol-block-containing shells. AB - We have examined the impact on biological systems of some newly synthesised polyoxyethylene polymers using in vitro assays. Toxicity was tested by the 3-[4,5 dimethylthiazole-2-yl]-2,5 diphenyltetrazolium bromide (MTT) assay, haemolysis was assessed, and an ethidium bromide (EB) assay was used to study interactions between the polymers and DNA. All the assay data showed that the polymers are biocompatible. No differences were found between generations (i.e. macromolecule sizes). The results encourage continuing studies on the clinical use of these molecules as drug carriers. PMID- 16378737 TI - Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata. AB - To date enormous attempts have been devoted to improve Taxol production exploiting various methodologies from bioprocess engineering to biotechnological and synthetic approaches. We have developed a 2-stage suspension cell culture of Taxus baccata L. using modified B5 medium in order to improve cell growth as well as productivity. After callus induction and cell line selection, B5 medium was supplemented with vanadyl sulfate (0.1 mg/l), silver nitrate (0.3 mg/l) and cobalt chloride (0.25 mg/l) at the first day of stage I culture to maximize cell growth. This medium was further supplemented with sucrose (1%) and ammonium citrate (50 mg/l) on day 10 and sucrose (1%) and phenylalanine (0.1 mM) on day 20 (i.e., biomass growth medium). At stage II (day 25), two different concentrations of several elicitors such as methyl jasmonate (10 or 20 mg/l), salicylic acid (50 or 100 mg/l) and fungal elicitor (25 or 50 mg/l) were added to the biomass growth medium with the aim of improving cellular productivity. For morphological analysis, microscopic inspection was carried out during cultivation. Cell associated and extracellular amount of Taxol were detected and measured using HPLC methodology. At stage I, overall Taxol amount of biomass growth medium was 13.75 mg/l (i.e., 5.6-fold higher than that of untreated B5 control). At stage II, treated cells with methyl jasmonate (10 mg/l), salicylic acid (100 mg/l) and fungal elicitor (25 mg/l) produced the highest amount of Taxol (39.5 mg/l), which is 16-fold higher than that of untreated B5 control (2.45 mg/l). Microscopic analyses of Taxus cells in suspension cultures showed various positional auto fluorescence showing direct correlation with Taxol production. Our studies revealed that intervallic supplementation of B5 medium with combination of biomass growth factors at stage I and mixture of elicitors at stage II could significantly increase Taxol production. Thus, we suggest that the exploitation of this methodology may improve the production of Taxol since demands for Taxol pharmaceuticals are increasingly growing and resource paucities have limited its direct harvesting from Taxus trees. PMID- 16378738 TI - Function of the F1-motor (F1-ATPase) of ATP synthase by apolar-polar repulsion through internal interfacial water. AB - THESIS: Within the structurally-confined internal aqueous cavity of the F1-motor of ATP synthase, function results from free energy changes that shift the balance between interfacial charge hydration and interfacial hydrophobic hydration. TRANSITION STATE DESCRIPTION: At the beta-P end of ADP x Mg occurs an inorganic phosphate, P(i). This P(i) resides at the base of a water-filled cleft that functions like an aperture to focus, into an aqueous chamber, a competition for hydration (an apolar-polar repulsion) between charged phosphate and hydrophobic surface of the gamma-rotor. Two means available for the phosphate and the hydrophobic surface to improve their hydration free energies are physically to separate by rotation of the gamma-rotor or chemically to combine P(i) with ADP to form less charged ATP. This proposal derives from calculated changes in Gibbs free energy for hydrophobic association of amino acid side chains and chemical modifications thereof and from experimentally demonstrated water-mediated repulsion between hydrophobic and charged sites that resulted from extensive studies on designed elastic-contractile model proteins. PMID- 16378739 TI - Overexpressed LEF-1 proteins display different nuclear localization patterns of beta-catenin in normal versus tumor cells. AB - Beta-catenin not only plays a role in cadherin-dependent cell adhesion, but also interacts with T-cell factor (TCF)/lymphoid enhancer factor-1 (LEF-1) to affect gene expression. In this report, we describe the effects of exogenous LEF-1 and of treatment with leptomycin B (LMB), a specific inhibitor of CRM1-medicated nuclear export, on the nuclear localization and export of beta-catenin. Normal epithelial cells overexpressing LEF-1 accumulate nuclear beta-catenin in a LEF-1 concentration-dependent manner. Nuclear beta-catenin, once imported from the cytoplasm, is rapidly removed from the nucleus. Treatment with LMB results in dramatic retention of nuclear beta-catenin in normal epithelial cells transfected with LEF-1, and this effect is intensified by treatment of N-Acetyl-leucyl-leucyl norleucinal together with LMB. Colon carcinoma cells containing an adenomatous polyposis coli mutation retain significant amounts of LEF-1 induced nuclear beta catenin considerably after the time-point when beta-catenin disappears from the nuclei of LEF-1 transfected normal epithelial cells. beta-Catenin binds directly to CRM1, and overexpression of CRM1 reduces nuclear beta-catenin-mediated transactivation function. PMID- 16378740 TI - Making an escape: development and function of the Drosophila giant fibre system. AB - Flies escape danger by jumping into the air and flying away. The giant fibre system (GFS) is the neural circuit that mediates this simple behavioural response to visual stimuli. The sensory signal is received by the giant fibre and relayed to the leg and wing muscle motorneurons. Many of the neurons in the Drosophila GFS are uniquely identifiable and amenable to cell biological, electrophysiological and genetic studies. Here we review the anatomy and development of this system and highlight its utility for studying many aspects of nervous system biology ranging from neural development and synaptic plasticity to the aetiology of neural disorder. PMID- 16378741 TI - A discoidal lipoprotein from the coelomic fluid of the polychaete Nereis virens. AB - A discoidal lipoprotein was isolated from the coelomic fluid of the polychaete, Nereis virens, by density gradient centrifugation. The lipoprotein was present in both sexes and moved as a uniform band in an agarose gel. The average diameter of the lipoprotein particles determined by electron microscopy was 42 nm with a thickness of 10 nm. SDS electrophoresis showed two apoprotein subunits with molecular masses of 247 and 85 kDa, respectively. In lectin blots, both apoproteins were reactive with Concanavalin A indicating the presence of N glycans. The small subunit was also reactive with peanut lectin, indicating additional O-glycosylation. The total lipid content was 48% and consisted mainly of phospholipids and some diglycerides as judged by thin layer chromatography. The estimated native molecular mass of N. virens lipoprotein ( approximately 675 kDa) lies in the range of vertebrate high-density lipoprotein and insect lipophorins. The size of the apoproteins is similar to those found in insects, while the composition of the lipid fraction is more similar to that of crustacean lipoproteins. PMID- 16378742 TI - A structure-function study of MID1 mutations associated with a mild Opitz phenotype. AB - The X-linked form of Opitz syndrome (OS) affects midline structures and produces a characteristic, but heterogeneous, phenotype that may include severe mental retardation, hypertelorism, broad nasal bridge, widow's peak, cleft lip/cleft palate, congenital heart disease, laryngotracheal defects, and hypospadias. The MID1 gene was implicated in OS by linkage to Xp22. It encodes a 667 amino acid protein that contains a RING finger motif, two B-box zinc fingers, a coiled-coil, a fibronectin type III (FNIII) domain, and a B30.2 domain. Several mutations in MID1 are associated with severe OS. Here, we describe an intelligent male with a milder phenotype characterized by hypertelorism, broad nasal bridge, widow's peak, mild hypospadias, pectus excavatum, and a surgically corrected tracheo esophageal fistula. He has an above average intelligence and no cleft lip/palate or heart disease. We identified a novel mutation in MID1 (P441L) which is in exon 8 and functionally associated with the FNIII domain. While OS phenotypes have been attributed to mutations in the C-terminal part of MID1, little is currently known about the structure-function relationships of MID1 mutations, and how they affect phenotype. We find from a literature review that missense mutations within the FNIII domain of MID1 are associated with a milder presentation of OS than missense mutations elsewhere in MID1. All truncating mutations (frameshift, insertions/deletions) lead to severe OS. We used homology analysis of the MID1 FNIII domain to investigate structure-function changes caused by our missense mutation. This and other missense mutations probably cause disruption of protein protein interactions, either within MID1 or between MID1 and other proteins. We correlate these protein structure-function findings to the absence of CNS or palatal changes and conclude that the FNIII domain of the MID1 protein may be involved in midline differentiation after neural tube and palatal structures are completed. PMID- 16378743 TI - Childhood ataxia with CNS hypomyelination/vanishing white matter disease--a common leukodystrophy caused by abnormal control of protein synthesis. AB - Mutations in eukaryotic initiation factor 2B (eIF2B) cause one of the most common leukodystrophies, childhood ataxia with CNS hypomyelination/vanishing white matter disease or CACH/VWM. Patients may develop a wide spectrum of neurological abnormalities from prenatal-onset white matter disease to juvenile or adult-onset ataxia and dementia, sometimes with ovarian insufficiency. The pattern of diffuse white matter abnormalities on MRI of the head is often diagnostic. Neuropathological abnormalities indicate a unique and selective disruption of oligodendrocytes and astrocytes with sparing of neurons. Marked decrease of asialo-transferrin in cerebrospinal fluid is the only biochemical abnormality identified thus far. Eukaryotic translation initiation factor 2B (eIF2B) mutations cause a decrease in guanine nucleotide exchange activity on eIF2-GDP, resulting in increased susceptibility to stress and enhanced ATF4 expression during endoplasmic reticulum stress. eIF2B mutations are speculated to lead to increased susceptibility to various physiological stress conditions. Future research will be directed towards understanding why abnormal control of protein translation predominantly affects brain glial cells. PMID- 16378744 TI - Mucopolysaccharidosis type IV: N-acetylgalactosamine-6-sulfatase mutations in Tunisian patients. AB - Mucopolysaccharidosis type IVA (MPS IVA; OMIM #253000) or Morquio A syndrome is an autosomal recessive inborn error resulting from the deficient activity of the lysosomal enzyme, N-acetylgalactosamine-6-sulfatase (GALNS), and the progressive lysosomal accumulation of sulfated glycosaminoglycans. Clinically, the severe form of this lysosomal storage disease is characterized by a characteristic severe bone dysplasia and normal intelligence. To date, a variety of mutations have been associated with the severe MPS IVA phenotype. Here, we report the GALNS mutations in six severe MPS IVA patients from four unrelated Tunisian families. For mutation detection, each of the 14 exons and adjacent intron-exon junctions of the GALNS gene were sequenced after PCR-amplification from genomic DNA. Two novel mutations were identified: a G to A transition in the conserved 5' donor splice site of intron 1 (GACgt-->GACat: designated IVS1(+1g-->a)) and a G to C transversion in codon 66 of exon 2 predicting a glycine to arginine substitution (G66R). The IVS1(+1g-->a) mutation was homozygous in five similarly affected patients from three presumably unrelated families, but haplotype analysis suggested a common ancestor. The affected patient in the fourth family was homozygous for the G66R mutation. These are the first GALNS mutations causing severe MPS IVA disease identified in Tunisia. These molecular findings provide genotype/phenotype correlations, and permit accurate carrier detection, prenatal diagnosis, and counseling for MPS IVA disease in Tunisia where first cousin consanguineous mating remains frequent. PMID- 16378745 TI - Using urea dilution to standardise cellular and non-cellular components of pleural and bronchoalveolar lavage (BAL) fluids in the cat. AB - A technique to standardise the analysis of cellular and non-cellular components in epithelial lining fluid (ELF) collected during saline lavage of pulmonary and pleural cavities was developed using the urea dilution method. Bronchoalveolar lavage (BAL) and pleural lavage (PL) fluids were collected from 12 clinically healthy cats. Total and differential cell counts in BAL fluid were within normal ranges for the cat, while cell counts in PL fluid were assumed to be normal based on clinical health during examination, auscultation and lactate dehydrogenase (LDH) activities being comparable with other species. The major clinical implication of this study was that nucleated cell counts within feline ELF could not be predicted from analysis of lavage fluid which suggests that calculation of the proportion of ELF in lavage fluid by the urea dilution method may be necessary to avoid misdiagnosis of health or disease in pulmonary or pleural cavities. PMID- 16378746 TI - Adverse effects of EMLA (lidocaine/prilocaine) cream and efficacy for the placement of jugular catheters in hospitalized cats. AB - EMLA is a lidocaine/prilocaine cream used for topical analgesia in human pediatric patients. The purpose of this study was to establish the safety of EMLA in clinically ill cats, to measure systemic absorption and to determine whether EMLA reduced the need for sedation for the placement of jugular catheters. Thirty one cats were randomized to either a placebo or EMLA cream group. Cream was applied to a 10 cm(2) area over the jugular vein, with 1h of occlusive dressing. Neither anesthetic was systemically absorbed in any cat, and no adverse clinical signs were observed. Struggling during catheter placement was less in the EMLA treated cats compared to placebo, but did not reach significance (P = 0.06). Jugular catheters were successfully placed in 60% of EMLA-treated cats and 38% of placebo cats; this difference was not statistically significant and may not justify the added steps of EMLA cream administration for this purpose. However, EMLA does appear to be safe in clinically ill cats, and may be useful for other applications such as for skin mass removal or repeated venepuncture. PMID- 16378747 TI - Contribution of adipocyte-derived factors to beta-cell dysfunction in diabetes. AB - In addition to serving as an energy reservoir, the adipocyte has been characterized as an endocrine cell, secreting many bioactive factors which influence energy homeostasis. Being overweight, with excessive adipose tissue, is considered to be part of the pathogenesis of type 2 diabetes. Insulin resistance and beta-cell dysfunction are two major pathophysiological changes seen in type 2 diabetes. In addition to inducing insulin resistance in insulin-responsive tissues, adipocyte-derived factors play an important role in the pathogenesis of beta-cell dysfunction. Leptin, free fatty acids, adiponectin, tumor necrosis factor-alpha and interleukin-6 are all produced and secreted by adipocytes, and may directly influence aspects of beta-cell function, including insulin synthesis and secretion, insulin cell survival and apoptosis. During the progression from normal weight to obesity and on to overt diabetes, the adipocyte-derived factors contribute to the occurrence and development of beta-cell dysfunction and type 2 diabetes. PMID- 16378748 TI - Overexpression of the novel human gene, nuclear apoptosis-inducing factor 1, induces apoptosis. AB - Apoptosis is a genetically determined cell suicidal program that plays critical roles in many physiological and pathological processes. In this study, we report the cloning and characterization of a novel human gene, nuclear apoptosis inducing factor 1 (NAIF1), overexpression of which induces apoptosis in cells. Human NAIF1 is located on chromosome 9q34.11 and encodes 327 amino acids with a homeodomain-like region and two nuclear localization signals at its N-terminal region. NAIF1 is conserved across diverse species, including human, mouse, crab eating macaque, dog, chicken and frog, and shares no obvious homology to any known genes or proteins. Northern blot analysis revealed wide expression of NAIF1 mRNA throughout human tissues. NAIF1 was predominantly localized in the nucleus. Overexpression of NAIF1 inhibited cell growth and induced apoptosis. Furthermore, NAIF1 transfection caused both decreases in mitochondrial membrane potential and caspase-3 activation. In summary, NAIF1 is a nuclear protein that induces apoptosis when overexpressed. PMID- 16378749 TI - Understanding the architecture of language: the possible role of neurology. AB - Culicover and Jackendoff have recently described an approach to language representation where semantic structure works, alongside syntax, as a generative system with its own structure and principles of composition. Well-known neurological observations support this view. They show that in the presence of a syntactic impairment, comprehension can take place but only if the sentence's semantic structure is rich enough. This would suggest the existence of syntax independent semantic combinatorial mechanisms, as Culicover and Jackendoff's model proposes. PMID- 16378750 TI - The role of foetal red blood cells in protecting cultured lymphocytes against diepoxybutane-induced chromosome breaks. AB - Diepoxybutane (DEB) is an established mutagen that induces chromosome damage following in vitro treatment of peripheral blood lymphocytes. It is widely used to identify patients with Fanconi Anemia (FA), a clinical situation that is characterized, besides the hypersensitivity to DEB, by an elevated foetal haemoglobin (HbF) content in the peripheral blood. In a previous study, we showed that red blood cells (RBC) from normal individuals can protect cultured lymphocytes against chromosomal breaks induced by DEB and demonstrated the particular role of haemoglobin in the protective effect. In the present work, we studied the influence of RBC extracted from umbilical cord blood of neonates (F cells) on the frequency of DEB-induced chromosome breaks in lymphocyte cultures from normal individuals. Simultaneously, we determined individual GSTT1 and GSTM1 genotypes and the activity of Pi-class glutathione S-transferase (GSTP), catalase and superoxide dismutase (SOD) in adult and foetal RBC. Our results show that F cells, in comparison with adult RBC, elicit a better protection of cultured lymphocytes from normal individuals against chromosome breaks induced by DEB. Variability in the protective effect among RBC from different individuals was observed; we confirmed that the GSTT1 genotype modulates this inter-individual variability, but it is not sufficient to explain all of the protective effect of F cells. Our results suggest that the increased protective effect of F cells can be, at least in part, correlated with an increase in the activity of glutathione S-transferase, catalase and superoxide dismutase, in particular Cu/Zn SOD, in F cells compared with adult RBC. PMID- 16378751 TI - Mitochondrial DNA mutation frequencies in experimentally irradiated compost worms, Eisenia fetida. AB - The compost worm Eisenia fetida is routinely used in ecotoxicological studies. A standard assay to assess genetic damage in this species would be extremely valuable. Since mitochondrial DNA (mtDNA) is known to exhibit an increased mutation rate following exposure to ionising radiation we assessed the validity of a mtDNA-based assay for measuring increases in mutation rate in laboratory irradiated compost worms. To this end the mutation frequency in the mtDNA of the compost worm E. fetida was quantified following in vivo gamma-irradiation of adult worms in three dose groups. Five adult worms exposed to 1.4 mGy/h for 55 days (total dose 1.85 Gy), five adult worms exposed to 8.5 mGy/h for 55 days (total dose 11.22 Gy) and five adult control worms were used to assess the effect of irradiation on mtDNA mutation induction. DNA samples extracted from irradiated adult worms were used in high-fidelity PCR of a 486 bp region of mtDNA spanning the ATPase 8 gene, chosen for its high spontaneous mutation rate. PCR products were cloned and sequenced to identify mutations, with 89-102 clones successfully sequenced per individual. A significant elevation in mtDNA mutation frequency (p=0.032) was seen in worms exposed at the higher dose rate (8.5 mGy/h, total dose 11.22 Gy; mutation frequency 27.98+/-4.85 x 10(-5)mutations/bp) in comparison to controls (mutation frequency 12.68+/-3.06 x 10(-5)mutations/bp), but no elevation in mutation frequency (p=0.764) was seen for the lower dose rate (1.4 mGy/h, total dose 1.85 Gy; mutation frequency 13.74+/-1.29 x 10( 5)mutations/bp) compared with controls. This indicates that although the technique has the potential to detect an elevation in mutation frequency, it does not have sufficient sensitivity at the doses likely to be encountered in environmental monitoring scenarios. PMID- 16378752 TI - New Pudicinae (Trichostrongylina, Heligmosomoidea), Pudica ctenomydis n. sp. parasite of Ctenomys talarum (Rodentia: Octodontidae) from Argentina. AB - A total of 138 nematodes were found in the small intestine of Ctenomys talarum (Octodontidae) from Mar de Cobo, Argentina. A new nematode species, Pudica ctenomydis n. sp., is described. The new species more closely resembles P. pujoli Durette-Desset, 1990, parasite of Microcavia niata Thomas, from Bolivia. It can be distinguished from P. pujoli by the number of ridges and characteristics of the synlophe, the spicular morphology, differences in length between rays 9 and 10, and by the presence of a symmetrical caudal bursa and a cuticular expansion surrounding the body between vulva and anus in females. PMID- 16378753 TI - Intense upconversion luminescence and effect of local environment for Tm3+/Yb3+ co-doped novel TeO2-BiCl3 glass system. AB - We present the results of a study that uses theoretical and experimental methods to investigate the characteristics of the upconversion luminescence of Tm3+/Yb3+ codoped TeO2-BiCl3 glass system as a function of the BiCl3 fraction. These glasses are potentially important in the design of upconversion fiber lasers. Effect of local environment around Tm3+ on upconversion fluorescence intensity was analyzed by theoretical calculations. The structure and spectroscopic properties were investigated in the experiments by measuring the Raman spectra, IR transmission spectra, and absorption and fluorescence intensities at room temperature. The results indicate that blue luminescence quantum efficiency increases with increasing BiCl3 content from 10 to 60 mol%, which were interpreted by the increase of asymmetry of glass structure, decrease of phonon energy and removing of OH- groups. PMID- 16378754 TI - Expression of mouse Coiled-coil-DIX1 (Ccd1), a positive regulator of Wnt signaling, during embryonic development. AB - Wnt signaling plays an important role in cell growth, differentiation, polarity formation, and neural development. We have recently identified the Coiled-coil DIX1 (Ccd1) gene encoding a third type of a DIX domain-containing protein. Ccd1 forms homomeric and heteromeric complexes with Dishevelled and Axin, and positively regulates the Wnt/beta-catenin pathway. Here, we examined the spatiotemporal expression pattern of Ccd1 mRNA in mouse embryos from embryonic day 6.5 (E6.5) to E17.5 by in situ hybridization. Ccd1 expression was detected in the node region in gastrula embryos, in the cephalic mesenchyme and tail bud at E8.5, and in the branchial arch and forelimb bud at E9.5. In the central nervous system, Ccd1 expression began and persisted in the regions where the neurons differentiated, so that it was observed throughout the brain and spinal cord at E17.5. Ccd1 expression was also strong in the peripheral nervous system, including sensory cranial ganglia (trigeminal, facial, and vestibulocochlear ganglia), dorsal root ganglia, and autonomic ganglia (sympathetic ganglia, celiac ganglion, and hypogastric plexus). Ccd1 was detected in the sensory organs, such as the inner nuclear layer of the neural retina, saccule and cochlea of the inner ear, and nasal epithelium. Outside the nervous system, Ccd1 mRNA was observed in the cartilage, tongue, lung bud, stomach, and gonad at E12.5-E14.5, and in the tooth bud, bronchial epithelium, and kidney at E17.5. Taken together, these findings demonstrate that Ccd1 expression is observed in all the neurons in the nervous system, closely associated with neural crest-derived tissues, and largely overlapping with the regions where several Wnt genes are reported to play a role. PMID- 16378755 TI - The LRR and Ig domain-containing membrane protein SST273 is expressed on motoneurons. AB - We performed signal sequence trap cDNA cloning using a cDNA library of enriched chicken embryonic spinal motoneurons and identified a novel transmembrane molecule, SST273 (Signal Sequence Trap clone 273). SST273 has five leucine-rich repeats (LRRs) and one immunoglobulin (Ig) domain in its extracellular domain. The human (KIAA1465) and mouse (BC059068) homologues of SST273 were already cloned, but had not yet been analyzed. The amino acid homologies between chick SST273 and human or mouse homologue are 52.1 and 51.5%, respectively. SST273 mRNA and its protein product were uniquely expressed in the embryonic spinal and cranial motoneurons at early developmental stages. PMID- 16378756 TI - The expression pattern of the Drosophila vesicular glutamate transporter: a marker protein for motoneurons and glutamatergic centers in the brain. AB - To determine the functions of genes in distinct tissues during the development of Drosophila, it is often desirable to have genetic tools for targeted gene expression in restricted subsets of cells. Here, we report the identification of the enhancer trap line OK371-Gal4, which is expressed in a defined subset of neurons from embryonic stage 15 to adulthood. In the ventral nerve chord, it is expressed almost exclusively in motoneurons and in the brain in a limited number of neuronal clusters. The OK371 enhancer trap element is inserted in the proximity of the annotated gene CG9887, which encodes a Drosophila vesicular glutamate transporter (DVGLUT). In situ hybridization experiments using antisense probes against the mRNAs of DVGLUT and neighboring genes confirm that OK371-Gal4 detects an enhancer of DVGLUT. DVGLUT-specific antibodies detect its expression in identifiable motoneurons, which are known to be glutamatergic in Drosophila. DVGLUT initially appears in small cytoplasmic punctae in the somata of these motoneurons. As development proceeds, DVGLUT-positive particles are transported along motor axons and become concentrated at neuromuscular junctions (NMJs), where they colocalize with the synaptic vesicle marker synaptotagmin. We find that the DVGLUT-specific antibodies are valuable tools for the identification of motoneurons and other glutamatergic neurons. In addition, the OK371-Gal4 line can be used for the targeted expression of any gene in these cells. Given that vesicular glutamate transporters are essential for the uptake of the neurotransmitter glutamate into synaptic vesicles these tools provide a means to test gene function in these functionally important neurons. PMID- 16378757 TI - Cloning and embryonic expression of zebrafish PLAG genes. AB - PLAG transcription factors play important roles in oncogenesis. To date three members of this subfamily of zinc finger proteins have been identified in humans and mice: PLAG1, PLAGL1 and PLAGL2. In this study, we identified zebrafish orthologs of PLAG1 and PLAGL2 and a novel member of this family, PLAGX. We examined the temporal expression of these three genes by quantitative real time RT-PCR and found that all three genes are maternally provided, expressed at low level during early somitogenesis and, during late somitogenesis and beyond, PLAG expression increases to reach a plateau level around 5 dpf. Whole mount in situ experiments revealed that PLAG1, PLAGL2 and PLAGX display a similar pattern of expression characterized by a low ubiquitous expression overcame by high expression in some restricted compartments such as the ventricular zone of the brain, the pectoral fin buds, the developing pharyngeal arches and the axial vasculature. We show that this pattern resembles the one observed for the proliferative marker PCNA, suggesting that the PLAG genes are expressed more strongly in zones of active proliferation. This hypothesis was proven for the ventricular zone shown to be a highly proliferative zone using the anti phosphohistone H3 antibody that detects cells in mitosis. PMID- 16378758 TI - Expression of coiled-coil protein 1, a novel gene downstream of FGF2, in the developing brain. AB - Fibroblast growth factor 2 (FGF2) plays an important role in cortical development. However, the genes downstream of FGF2 that mediate its effect are largely unknown. We have performed a microarray screening of genes regulated by FGF2 using primary cortical neuron culture derived from embryonic day 14.5 (E14.5) mouse forebrains. In this study, we have analysed a previously uncharacterised gene encoding a 180-amino acid protein, hereby named 'coiled-coil protein 1 (ccp1)', that showed a modest up-regulation upon FGF2 stimulation. Northern blots and RT-PCR showed specific expression of ccp1 in multiple tissues including adult and embryonic brains. In situ hybridizations revealed that ccp1 was expressed in the cortical plate between Reelin and Tbr1-positive layers in the dorsal cortex at E15.5. Furthermore, the expression pattern of ccp1 at E13.5 E14.5 reflected some of the aspects of tangential migration of cortical progenitors during the early phase. We observed that the expressed ccp1 protein was localised to endo/lysosomal compartment in the cell body as well as to vesicles present in the processes of primary cortical neurons and oligodendrocyte cell line. PMID- 16378759 TI - SAGE reveals expression of Wnt signalling pathway members during mouse prostate development. AB - To identify genes and pathways not previously implicated in the mesenchymal epithelial (M/E) interactions that are critical for normal mouse prostate development, we constructed six serial analysis of gene expression (SAGE) libraries. Bioinformatic analyses revealed expression of various members of numerous signalling pathways and the differential expression of several members of the wingless-related MMTV integration site (Wnt) signalling pathway. This pathway has not been previously implicated in prostate development thus expression of selected Wnt pathway members in the developing prostate was confirmed by RT-qPCR. Of particular interest, an antagonist of the Wnt pathway, secreted frizzled related protein 2 (Sfrp2), was highly expressed in the early prostate libraries and down regulated at later developmental stages. The expression levels of four Wnt ligands reported to interact with Sfrp2 were, therefore, examined by RT-qPCR. We found that only Wnt4 transcripts were detectable in the developing prostate. Expression of Sfrp2 was validated using RT qPCR and localization of Sfrp2 transcripts and protein was carried out using in situ hybridization and immunofluorescence, respectively. These studies provide the first evidence that Wnt pathway members are expressed in the developing prostate. Functional analyses are now required to establish the biological significance of this observation. PMID- 16378760 TI - Tissue-specific Spred-2 promoter activity characterized by a gene trap approach. AB - Spreds (Sprouty-related proteins with an Ena/Vasodilator-stimulated phosphoprotein homology-1 domain) are a new protein family inhibiting the mitogen activated protein kinase (MAPK) signaling pathway. Different RNA and protein studies already revealed an almost ubiquitous Spred-2 expression pattern. But until now, only few data were available on the in situ Spred-2 promoter activity. Here, we show a detailed in situ analysis of a mouse strain with a trapped Spred 2 gene, bringing a beta-galactosidase and neomycin fusion gene (beta-geo) under the control of the endogenous Spred-2 promoter. This allowed us to monitor Spred 2 promoter activity in practically every organ and their corresponding sub compartments. X-Gal staining of newborn and adult mice revealed a nearly congruent Spred-2 promoter activity pattern. Our detailed data provide information for further studies of the still enigmatic physiological functions of Spred-2 in various organs by identifying the tissues with strong Spred-2 promoter activity. PMID- 16378761 TI - Cloning and expression pattern of the Xenopus erythropoietin receptor. AB - Cytokine signaling plays an important role in the survival and differentiation of vertebrate hematopoietic cells. In red blood cells, erythropoietin is a key component of the differentiation program and maintains the homeostasis of the erythroid compartment. In the adult, anemia stimulates high levels of circulating erythropoietin that drives erythropoiesis to restore normal levels of red blood cells in circulation. Erythropoietin activates the erythropoietin receptor on immature red blood cell precursors to promote their survival and differentiation. Although extensively studied in mammalian systems, a complete understanding of the function of the erythropoietin receptor during primitive erythropoiesis has been lacking. To address this problem, we have cloned the Xenopus laevis erythropoietin receptor in order to further understand the development of primitive erythropoiesis. The amphibian erythropoietin receptor shares 33% amino acid sequence identity with the mammalian erythropoietin receptors and contains the conserved extracellular ligand binding and fibronectin domains, the WSXWS motif common to cytokine receptors, and several tyrosine phosphorylation sites located on the intracellular domain of the receptor. Expression of the erythropoietin receptor is first detected by in situ hybridization in the ventral blood island during tailbud stages. PMID- 16378762 TI - The expression patterns of deubiquitinating enzymes, USP22 and Usp22. AB - Deubiquitinating enzymes regulate a number of cellular mechanisms including pre implantation, growth and differentiation, oncogenesis, cell cycle progression, transcriptional activation, and signal transduction. In this study, we have identified a novel human deubiquitinating enzyme gene, USP22, and its mouse homologue, Usp22. They encode 525 amino acids (approximate MW: 60kDa) and contains Cys, Asp (I), His and Asp/Asn (II), the highly conserved domains of the UBP family of deubiquitinating enzymes. The biochemical assay revealed that they have deubiquitinating enzyme activity. Northern blot analysis for USP22 showed moderate expression in various organs including human heart and skeletal muscle, and weak expression in lung and liver. However, Usp22 is expressed strongly in brain and weakly in other organs. We investigated the expression level of Usp22 mRNA and the localization during implantation and early pregnancy by in situ hybridization. Interestingly, Northern blot analysis showed the strong expression of Usp22 between embryonic days E10.5 and E12.5. Whole mount in situ hybridization staining revealed that Usp22 was expressed in the midbrain, forebrain, hindbrain and dorsal root ganglia of embryos at E12.5. Embryos at E12.5 showed the pronounced expression of Usp22 during the early embryonic development, although its expression was not detectable in the gut, liver and heart. PMID- 16378763 TI - Cloning and expression pattern of lbx3, a novel chick homeobox gene. AB - The Drosophila melanogaster genes, ladybird early (lbe) and ladybird late (lbl), encode transcriptional regulators, which play an important role in neurogenesis, myogenesis and cardiogenesis. Here, we report an isolation of a novel ladybird family homeobox gene (lbx3) and its expression during chick development. The open reading frame of lbx3 encodes a predicted protein of 213 amino acids including a homeodomain, a PST motif and a nuclear localization signal. The homeodomain of lbx3 protein has 80% identities with the chick and mouse lbx1 homeodomains, and 75% identity with the mouse lbx2 homeodomain. Both lbx1 and lbx3 are expressed in prospective hypaxial myoblasts at cervical and limb level. In addition, lbx3 transcripts are detected in the medial dermomyotomal lips of somites of all axial levels at stage 23, but not detected in the neural tube. PMID- 16378764 TI - Characterization of immunoaffinity purified peptidoglycan-associated lipoprotein of Actinobacillus actinomycetemcomitans. AB - Peptidoglycan-associated lipoprotein (PAL) is a highly conserved structural outer membrane protein among Gram-negative bacteria. In some species, it is proinflammatory and released extracellularly. We purified a newly identified PAL (AaPAL) of a periodontal pathogen Actinobacillus actinomycetemcomitans by using AaPAL antipeptide antibodies coupled to immunoaffinity chromatography column. No protein impurities originating in A. actinomycetemcomitans were found in the final product. Sera from patients infected by A. actinomycetemcomitans recognized the purified AaPAL. The present purification method seems to be suitable for isolation of AaPAL and probably PALs of other bacterial species, and applicable in studies investigating proinflammatory mechanisms of A. actinomycetemcomitans. PMID- 16378765 TI - High-pressure liquid chromatographic analysis for identification of in vitro and in vivo metabolites of 4-phenethyl-5-[4-(1-(2-hydroxyethyl)-3,5-dimethyl-4 pyrazolylazo)phenyl]-2,4-dihydro-3H-1,2,4-triazole-3-thione in rats. AB - All azo colorants whose metabolism can liberate a carcinogenic arylamine, are suspected of having carcinogenic potential. Therefore, a new azo compound 4 phenethyl-5-[4-(1-(2-hydroxyethyl)-3,5-dimethyl-4-pyrazolylazo)phenyl]-2,4 dihydro-3H-1,2,4-triazole-3-thione (substrate) was prepared to investigate its in vitro and in vivo biotransformation in rats by HPLC. Chromatographic separation of substrate and its metabolites was performed using a Chromasil C(18) column. The mobile phase consisted of acetonitrile and water in a linear gradient system. From the biotransformation of this compound, the reduction metabolite 4-(2 phenethyl)-5-(4-aminophenyl)-2,4-dihydro-3H-1,2,4-triazole-3-thione was identified by comparing it to reference standard by HPLC-DAD. In the in vivo study, identification of the unknown peak which was the N-acetylation metabolite was confirmed by LC-MS spectrometry. Besides this, the azo compound was reduced to its corresponding amine in intestinal and cytosolic parts. In addition, oxidation of the methyl group and the phenyl ring, and reduction of azo group to hydrazo were identified in the cytosolic part using LC-MS. PMID- 16378766 TI - Quantitative analysis of trimethylsilyl derivative of hydroxyurea in plasma by gas chromatography-mass spectrometry. AB - Hydroxyurea is an antitumor drug widely used in the treatment of sickle cell disease. The drug has been analyzed in biological fluids by a number of high performance liquid chromatography (HPLC) methods. This paper describes a fast and highly reliable capillary gas chromatography-mass spectrometry (GC-MS) procedure that was developed for the detection and quantitation of hydroxyurea in plasma. The compound and its labeled internal standard were liquid extracted from plasma and derivatized with BSTFA before analysis. The detection limit of the assay was 0.078 microg/ml and the limit of quantitation was 0.313 microg/ml with linearity up to 500 microg/ml. Intra-day variation, as coefficient of variation (C.V., %) over the selected concentration range, was 0.3-8.7% and inter-day variation was 0.4-9.6%. PMID- 16378767 TI - Determination of endogenous glycosaminoglycans derived disaccharides in human plasma by HPLC: validation and application in a clinical study. AB - SB-424323 is a new, orally active anti-thrombotic agent presently in phase-II clinical development, with limited hemorrhagic risk and a unique mechanism of action involving the induction of glycosaminoglycans (GAGs) biosynthesis. The objective of the present study was to develop a simple and rapid high performance liquid chromatography (HPLC) method for determination of endogenous GAGs derived disaccharides in plasma samples from a phase-II clinical study of SB-424323. Sample preparation was a simple heat treatment of the diluted plasma followed by digestion of endogenous GAGs with chondroitinase ABC to yield unsaturated disaccharides, 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-D galactose (DeltaDi-0S), 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4 enepyranosyluronic acid)-4-O-sulfo-D-galactose (DeltaDi-4S), and 2-acetamido-2 deoxy-3-O-(beta-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose (DeltaDi 6S). These disaccharides were recovered and purified using centrifugal filtration through a filter with 3000 molecular weight cut-off along with externally added internal standard 2-acetamido-2-deoxy-3-O-(2-O-sulfo-beta-D-gluco-4 enepyranosyluronic acid)-D-galactose (DeltaDi-UA2S). A gradient reverse phase HPLC separation was developed on a Waters Symmetry C(18) column (4.6 mm x 150 mm, 5 microm) with a gradient mobile phase system consisting of 0.8 mM tetrabutylammonium hydrogen sulfate and 2mM sodium chloride and acetonitrile at a flow rate of 1.0 mL/min. The eluate was monitored with an ultraviolet detector set at 230 nm. Plasma standard curves were linear (r(2)> or =0.994) in the concentration range 1.0-20 microg/mL with a lower limit of quantification (LLOQ) of 1.0 microg/mL for each of the disaccharide. The mean measured quality control (QC) concentrations for the disaccharides deviated from the nominal concentrations in the range of -8.92 to 5.61% and -16.3 to 16.7%, for inter and intra-day, respectively. The inter and intra-day precision in the measurement of QC samples, were in the range of 3.21 to 18.2% relative standard deviation (R.S.D.) and 0.32 to 20.9% R.S.D., respectively. The inter and intra-day precision in the measurement of endogenous GAGs derived disaccharides in human control plasma, were in the range of 5.8 to 15.9% R.S.D. and 1.17 to 7.74% R.S.D., respectively. Stability of the processed samples was confirmed up to 48 h in the auto-sampler. The method is simple, reliable, and easily adaptable to analysis of large number of samples under logistics of a clinical study. The present method has been used to investigate the GAGs levels in the plasma of patients in a phase II clinical study of SB-424323. PMID- 16378769 TI - An in-flight reflection on safety and quality. PMID- 16378770 TI - Clinical opinion prevails over the pneumonia severity index. PMID- 16378771 TI - Cardiovascular risk factors in minorities. AB - Recent clinical trials have confirmed the value of intervention on major risk factors, particularly hypertension and hyperlipidemia, in preventing the progression and clinical sequelae of atherosclerosis. Less is known about the prevalence and impact of atherosclerosis risk factors in minorities. A review of recent literature reporting the prevalence of established and new predictors of atherosclerotic events in minority populations and the inclusion of minorities in clinical trials is presented. The prevalence of risk factors differs considerably in minority populations. The role of "premature" coronary death and the level of some risk factors, particularly obesity and blood pressure in African descendants and high triglycerides, low high-density lipoproteins, and diabetes in some Hispanics, is higher than in whites. With few exceptions, however, minorities have not been included in clinical trials in sufficient numbers to determine whether significant differences in the benefit of risk factor intervention exists. Prevalence of key risk factors differs among minority groups. Risk factor intervention should be pursued in minority groups but with the understanding that clinical trials have not ruled out the possibility of qualitative or quantitative differences in response rates among different groups. PMID- 16378772 TI - Progression of chronic kidney disease: can it be prevented or arrested? AB - Chronic kidney disease constitutes a highly prevalent health problem worldwide. Left untreated, it progresses inexorably to greater levels of severity at variable rates. The morbid impact of chronic kidney disease is heightened by its role as risk factor for cardiovascular disease. In the past two decades, considerable gains have been realized in retarding progression of chronic kidney disease by emphasizing blood pressure control and blockade of the renin angiotensin system. Notwithstanding, the therapeutic goal of preventing or arresting chronic kidney disease progression remains unfulfilled. Currently attainable rates of decrease in glomerular filtration rate remain at 2 to 8 mL/min/y depending on the underlying disease. It is now believed that to achieve optimal therapeutic targets (proteinuria of <500 mg/day and decrease in glomerular filtration rate of 1 mL/min/y, the average age-related decline) we must introduce novel strategies and a multifaceted approach to treatment that interrupts multiple mechanisms of progression. To this end, and wherever relevant, new approaches to cause-specific treatment must be applied, such as targeted immunosuppression, intensive glycemic control, gene therapy, and enzyme replacement therapy. Furthermore, in all chronic kidney disease, we must interfere more effectively with the multitude of common mechanisms of progression. Established or putative, such approaches include aggressive blood pressure control; advanced renin-angiotensin system blockade; cytokine modulation and antifibrotic therapy; aldosterone blockade; endothelin blockade, nitric oxide modulation and vasopeptidase inhibition; antioxidant therapy; statin therapy; glycosaminoglycan therapy; anemia therapy; dietary restrictions; lifestyle changes; and pharmacogenomic profiling. Such a concerted, multifaceted approach to management might indeed prevent or arrest progression of chronic kidney disease, or even achieve regression of chronic kidney disease. PMID- 16378773 TI - Managing patients with lower urinary tract symptoms suggestive of benign prostatic hyperplasia. AB - Many men with lower urinary tract symptoms suggestive of benign prostatic hyperplasia can be managed safely and effectively by primary care providers. After a basic evaluation to exclude other diseases that may cause lower urinary tract symptoms, quantifying the degree of symptoms and bother, and perhaps making an assessment of prostate size, the primary care provider can determine which men require immediate evaluation by a urologist and which men may choose among various treatment options, including watchful waiting and various single agent or combination medication strategies. Recent information about risk factors for disease progression has also helped to inform patient decisions on which treatment option is best for the individual patient. The purpose of this review is to provide primary care providers with an approach to the management of men with lower urinary tract symptoms suggestive of benign prostatic hyperplasia. PMID- 16378774 TI - Evaluation and treatment of Cushing's syndrome. AB - Cushing's syndrome results from sustained pathologic hypercortisolism caused by excessive corticotropin (ACTH) secretion by tumors in the pituitary gland (Cushing's disease, 70%) or elsewhere (15%), or by ACTH-independent cortisol secretion from adrenal tumors (15%). The clinical features are variable, and no single pattern is seen in all patients. Those features most specific for Cushing's syndrome include abnormal fat distribution, particularly in the supraclavicular and temporal fossae, proximal muscle weakness, wide purple striae, and decreased linear growth with continued weight gain in a child. Patients with characteristics of glucocorticoid excess should be screened with measurements of saliva or urine cortisol or dexamethasone suppression testing. The diagnosis of Cushing's syndrome should be followed by the measurement of plasma ACTH concentration to determine whether the hypercortisolism is ACTH independent. In ACTH-dependent patients, bilateral inferior petrosal sinus sampling with measurement of ACTH before and after administration of ACTH releasing hormone most accurately distinguishes pituitary from ectopic ACTH secretion. Surgical resection of tumor is the optimal treatment for all forms of Cushing's syndrome; bilateral adrenalectomy, medical treatment, or radiotherapy are sought in inoperable or recurrent cases. The medical treatment of choice is ketoconazole. The prognosis is better for Cushing's disease and benign adrenal causes of Cushing's syndrome than adrenocortical cancer and malignant ACTH producing tumors. PMID- 16378775 TI - A pressing situation. PMID- 16378776 TI - What causes clubbing? PMID- 16378777 TI - Evolution of a T wave. PMID- 16378778 TI - An unusual cause of acute abdomen. PMID- 16378779 TI - Low-risk patients admitted with community-acquired pneumonia. AB - PURPOSE: To describe the natural history of community-acquired pneumonia in the subset of a large cohort of patients at low risk for mortality who were admitted to the hospital. METHODS: Prospective observational study of all patients at low risk for mortality (risk classes I and II) who presented to 6 hospitals and 1 emergency department in Edmonton, Alberta, Canada with a diagnosis of possible community-acquired pneumonia from November 15, 2000, to November 14, 2002. RESULTS: A total of 586/3065 (19.1%) low-risk patients (Fine criteria) were admitted, 48.4% of whom stayed more than 5 days. Multivariate analysis revealed that patients who were admitted were more likely to be female, to have presented at Site B, which serves an inner city population, to have diminished premorbid functional status, to have comorbidities likely to be made worse by pneumonia (chronic obstructive pulmonary disease, asthma, heart disease, inflammatory bowel disease), and to suffer from substance abuse or psychiatric illness. A respiratory rate of >/=28 breaths per minute, and symptoms of shaking chills, shortness of breath, nausea or diarrhea were the remaining factors predicting admission. Nineteen percent of the patients suffered one or more complications, the most serious of which was progression of the pneumonia, resulting in respiratory failure necessitating mechanical ventilation in 2.4% and empyema in 1.4%. Four patients had lung cancer, and 1 had cancer of the vocal cords. Thirty one percent of those who were admitted were still unable to eat or drink enough to maintain hydration by hospital day 5 or on discharge day. CONCLUSIONS: One in 5 patients at low risk for mortality were admitted to the hospital and half stayed more than 5 days; 19% suffered 1 or more complications. Our data emphasize the need for better rules to guide the admission decision and the importance of physician judgment in this decision. PMID- 16378780 TI - Airway obstruction in never smokers: results from the Third National Health and Nutrition Examination Survey. AB - PURPOSE: Chronic obstructive pulmonary disease (COPD) is usually described as a disease of cigarette smoking. COPD is rarely considered in persons with no smoking history except in the context of another exposure. Accordingly, the disease has not been well characterized in these "never smokers." METHODS: We evaluated airway obstruction (defined as forced expiratory volume in 1 second/forced vital capacity <0.70) in US adults aged 30 to 80 years interviewed in the Third National Health and Nutrition Examination Survey with valid spirometry who had never smoked. Previously described risk factors were examined for their association with obstruction in bivariate and multivariate analyses. RESULTS: Never smokers represented 42% of the Third National Health and Nutrition Examination Survey population aged 30 to 80 years, with obstruction prevalence of 91 per 1000. Never smokers accounted for 4.56 million cases of obstruction, or 23% of the total burden. Among these obstructed never smokers, 19% reported a prior diagnosis of asthma alone, and 12.5% reported COPD (alone or with asthma), leaving 68.5% with no prior respiratory diagnosis. After adjustment for other factors, higher rates of obstruction were significantly associated with increasing age, male sex, lower body mass index, and a history of allergies. CONCLUSIONS: Never smokers represent a significant proportion of airway obstruction in US adults. Only one fifth of obstruction in this group is explained by asthma. COPD may explain much of the remainder, although known risk factors were not explanatory in this dataset. Recommendations that lung health screening programs be limited to smokers should be reconsidered. PMID- 16378781 TI - Variability in the use of thromboprophylaxis and outcomes in critically ill medical patients. AB - PURPOSE: To describe practices for preventing venous thromboembolism in critically ill medical patients and to identify associations between prophylactic measures and survival. METHODS: We reviewed the records of all medical admissions to the intensive care units of a university hospital and an affiliated Veterans Affairs hospital over a 1-year period. We recorded patients' demographic characteristics, risk factors for venous thromboembolism, methods of prophylaxis, and in-hospital deaths. RESULTS: We identified 272 critically ill medical patients who received intensive care for at least 24 hours. Some form of prophylaxis was used in 205 patients (75%), including pharmacologic prophylaxis alone in 55 (20%), mechanical prophylaxis alone in 102 (38%), and both methods in 48 (18%). In-hospital mortality rates were 23% (24/103) for patients who received pharmacologic prophylaxis, and 36% (61/169) for those who received mechanical prophylaxis alone or no prophylaxis (P=.03). After adjusting for demographic characteristics, risk factors for thrombosis and severity of illness, the odds of death were 55% lower in patients who received pharmacologic prophylaxis (odds ratio [OR]=0.45; 95% confidence interval (CI): 0.22 to 0.93; P=.03). Similar results were obtained in propensity-adjusted and propensity-stratified analyses. Use of mechanical prophylaxis was not associated with survival (OR=0.88; 95% CI 0.44 to 1.77; P=.73). CONCLUSION: In this cohort of critically ill medical patients, pharmacologic but not mechanical thromboprophylaxis was associated with reduced risk of in-hospital death. This hypothesis must be tested in randomized trials. PMID- 16378782 TI - Pacemaker stress echocardiography predicts cardiac events in patients with permanent pacemaker. AB - PURPOSE: Noninvasive pacemaker stress echocardiography is a newly introduced method for the diagnosis of coronary artery disease in patients with a permanent pacemaker. The prognostic value of pacemaker stress echocardiography has not been studied. SUBJECTS AND METHODS: We studied 136 patients (mean age 64+/-12 years) with a permanent pacemaker who underwent pacemaker stress echocardiography for evaluation of coronary artery disease. All patients underwent pacemaker stress echocardiography by external programming (pacing heart rate up to ischemia or target heart rate). RESULTS: Thirty-one patients (23%) had normal study results. Ischemia was detected in 75 patients (55%). During a mean follow-up of 3.5+/-2.4 years, 35 deaths (26%) (20 the result of cardiac causes) and 2 nonfatal myocardial infarctions (1%) occurred. The annual cardiac death rate was 1.3% in patients without ischemia and 4.6% in patients with ischemia (P=.01). The annual all-cause mortality rate was 3.1% in patients without ischemia and 7% in patients with ischemia (P=.004). The presence of ischemia during pacemaker stress echocardiography was the strongest independent predictor of cardiac death (hazard ratio 4.1, confidence interval 1.2-14.5) and all-cause mortality (hazard ratio 2.7, confidence interval 1.2-6.0) in a multivariable model. CONCLUSION: Myocardial ischemia during pacemaker stress echocardiography is an independent predictor of cardiac death and all-cause mortality in patients with a permanent pacemaker. PMID- 16378783 TI - What do patients consider when making decisions about treatment for hepatitis C? AB - PURPOSE: There are few data describing decision-making in chronic hepatitis C infection from the patient's perspective. In this study, we sought to investigate the factors that influence patients' decisions as they consider treatment for hepatitis C infection. SUBJECTS: Consecutive patients attending outpatient liver clinics were recruited. Purposeful sampling was employed to include patients who were currently being treated or had recently been treated for chronic hepatitis C infection with pegylated-interferon and ribavirin as well as patients who had refused therapy. METHODS: We conducted focus groups until thematic saturation was reached. All focus groups were facilitated by the same PhD-level senior research scientist, and constant comparative methods were used to analyze the data. RESULTS: A total of 40 patients (80% male) participated in 8 focus groups. The factors influencing patients' decision-making that emerged most frequently during the focus groups were consideration of risk benefit tradeoffs, protected values, heuristics, participants' conceptualization of hepatitis C infection, social issues, and physicians' recommendations. CONCLUSION: Ideally, complex decision making is based on careful consideration of the tradeoffs related to available options. Our findings suggest that patients' treatment decisions are influenced by multiple factors besides the risks and benefits of interferon and ribavirin. By being aware of these factors physicians can improve decision-making in hepatitis C infection by 1) determining whether patients' decisions are biased by heuristics or protect values, 2) understanding how patients' conceptualization of their illness influences their attitudes toward therapy, and by 3) ensuring that patients understand that social responsibilities need not necessarily preclude treatment because therapy can be discontinued if adverse effects become intolerable. PMID- 16378784 TI - Quality of care in for-profit and not-for-profit health plans enrolling Medicare beneficiaries. AB - BACKGROUND: For-profit health plans now enroll the majority of Medicare beneficiaries who select managed care. Prior research has produced conflicting results about whether for-profit health plans provide lower quality of care. OBJECTIVE: The objective was to compare the quality of care delivered by for profit and not-for-profit health plans using Medicare Health Plan Employer Data and Information Set (HEDIS) clinical measures. RESEARCH DESIGN: This was an observational study comparing HEDIS scores in for-profit and not-for-profit health plans that enrolled Medicare beneficiaries in the United States during 1997. OUTCOME MEASURES: Outcome measures included health plan quality scores on each of 4 clinical services assessed by HEDIS: breast cancer screening, diabetic eye examination, beta-blocker medication after myocardial infarction, and follow up after hospitalization for mental illness. RESULTS: The quality of care was lower in for-profit health plans than not-for-profit health plans on all 4 of the HEDIS measures we studied (67.5% vs 74.8% for breast cancer screening, 43.7% vs 57.7% for diabetic eye examination, 63.1% vs 75.2% for beta-blocker medication after myocardial infarction, and 42.1% vs 60.4% for follow-up after hospitalization for mental illness). Adjustment for sociodemographic case-mix and health plan characteristics reduced but did not eliminate the differences, which remained statistically significant for 3 of the 4 measures (not beta-blocker medication after myocardial infarction). Different geographic locations of for profit and not-for-profit health plans did not explain these differences. CONCLUSION: By using standardized performance measures applied in a mandatory measurement program, we found that for-profit health plans provide lower quality of care than not-for-profit health plans. Special efforts to monitor and improve the quality of for-profit health plans may be warranted. PMID- 16378786 TI - The preoperative cardiovascular evaluation of the intermediate-risk patient: new data, changing strategies. AB - The intermediate-risk preoperative patient can be defined as a patient without severely symptomatic or unstable heart disease who, nonetheless, has clinical predictors of adverse perioperative cardiovascular events. Newer data have created an awareness of competing considerations in managing these patients. There is still debate about how to appropriately select patients for noninvasive cardiac testing, invasive coronary testing, coronary revascularization, beta blockers, or a combination of these. In this article, we review the evidence pertaining to these issues. We conclude that intermediate-risk preoperative patients are best managed by an approach that emphasizes the following points: intermediate-risk patients should be identified and risk stratified using a clinical tool (eg, the Revised Cardiac Risk Index); noninvasive cardiac testing should be reserved for those patients with multiple clinical predictors of risk or the presence of other modifying factors; preoperative coronary revascularization does not appear to reduce perioperative risk in patients with significant but stable coronary artery disease; and medical therapy should be optimized for these patients, including the application of beta-blockers in all intermediate-risk patients that do not have contraindications. PMID- 16378787 TI - The polyp detection rate of colonoscopy: a national study of Medicare beneficiaries. AB - National-level data on the proportion of colonoscopies in routine practice that result in polyp detection are limited. Our goal was to determine the diagnostic yield of colonoscopy among population subgroups. We reviewed Medicare claims data for all fee-for-service beneficiaries 65 years or older who underwent colonoscopy in 1999. With the use of diagnosis and procedure codes, the proportion of procedures with polyps was estimated. Three different criteria for polyp detection were used. Approximately 1.8 million colonoscopy procedures were identified. Depending on the specific criteria used, 23.9% to 35.7% of examinations noted polyps. The detection rate was highest in younger patients, men, and whites. The apparent yield of colonoscopy in terms of polyp detection is at least 24%. These data may be informative in predicting resource allocation for therapeutic procedures associated with colonoscopy. Future studies should determine the use and yield of serial colonoscopies for polyp surveillance. PMID- 16378788 TI - Hepatitis B virus infection: current status. AB - Hepatitis B virus currently infects more than 400 million people worldwide. Despite the availability of hepatitis B vaccine, the overall prevalence of hepatitis B virus infection has declined little in recent years. Hepatitis B virus causes liver injury by an immune response against the virus-infected liver cells and is not directly cytopathic, although immunosuppression appears to enhance replication and lead to direct cytotoxicity. The interplay of the host immune response and the virus's ability to replicate is a prime determinant of the likelihood of liver injury, its intensity, and progression to cirrhosis. A series of stages evolve in the life cycle of each patient's infection, with associated decreases in viral load at each successive stage. Viral mutations in the polymerase or the core gene affect replication and may enhance liver injury. Recently, genotypes have been identified that are linked to clinical outcomes, drug responses, and mutations. Four drugs (interferon alpha, lamivudine, adefovir, and entecavir) have been approved by the US Food and Drug Administration for treatment of hepatitis B virus; they effectively decrease replication and reduce inflammation and fibrosis. Treatment of hepatitis B virus in complex situations such as co-infection with human immunodeficiency virus or immunosuppressive therapy remains challenging. The use of hepatitis B vaccine has been shown to reduce the incidence of new infection in many regions. A decline in the prevalence of hepatitis B infection worldwide will require changes in high risk behavior and the wider use of vaccination. PMID- 16378789 TI - Lipid levels and bone mineral density. AB - PURPOSE: There has been considerable debate about the potential relationship between the use of statin lipid-lowering drugs and fracture risk; several observational studies suggest a protective effect but no randomized controlled trials have confirmed such a benefit. Because statins are given preferentially to persons with hyperlipidemia, if lipid levels were associated with bone mineral density, this could explain the discrepancy between epidemiological observations and randomized controlled trials. The aim of this study was to examine the relationship between lipid levels and bone mineral density. SUBJECTS AND METHODS: We included the 13592 participants in the National Health and Nutritional Examination Survey (NHANES) III who had bone mineral density and lipid levels measured; participants who reported the use of a lipid-lowering therapy were excluded. We examined the unadjusted bone mineral density across quintiles of total cholesterol, low-density lipoprotein (LDL), and high-density lipoprotein (HDL). We then constructed multivariable models, including age, sex, body mass index, and other potential confounders. RESULTS: In crude analyses, higher total cholesterol and LDL levels were associated with lower bone mineral densities (both P values for trend <.001), whereas higher HDL levels were associated with higher bone mineral densities (P value for trend <.001). However, in fully adjusted models, there was no significant relationship between total cholesterol, LDL, or HDL levels and bone mineral density (all P values for trend >.1). CONCLUSIONS: These results do not support a relationship between lipid levels and bone mineral density. PMID- 16378790 TI - Bridging the quality gap in diabetic hyperlipidemia: a practice-based intervention. AB - PURPOSE: Dyslipidemia treatment dramatically decreases coronary heart disease risk in diabetes, yet only a minority of these patients are screened or achieve optimal low-density lipoprotein (LDL) cholesterol levels. Our aim was to increase the percentage of diabetic patients in whom lipid management was achieved through electronic and direct educational detailing. METHODS: The study cohort comprised 884 diabetic patients at 12 primary care practices. Practice sites were randomized to one of three intervention groups: electronic educational detailing, direct (face-to-face) educational detailing, or control. Direct and electronic detailing were performed over a 12-month period. All sites were notified of our goal to enhance lipid testing among diabetic patients. Chart abstraction was performed 15 months after the start of the intervention. For the entire population (n=884), the proportion of patients with lipid testing was calculated, and changes from pre- to postintervention were compared across groups. We compared pre- and postintervention LDL-cholesterol changes between groups using least square means to account for site variation. RESULTS: Favorable provider actions increased significantly with the intervention (+22% compared with +6% in controls, P=.01). By logistic regression, electronic detailing increased the likelihood of lipid testing (odds ratio 3.0, confidence interval 1.6-5.7), as did direct detailing (odds ratio 1.8, confidence interval 0.9-3.7) in patients with no preintervention LDL test (n=432). Lipid testing tended to increase to a greater extent at intervention sites (+23% for the combination of electronic and direct detailing vs +11% for controls, P=.06). CONCLUSIONS: Brief educational detailing either through direct or electronic communication favorably impacts provider behavior regarding dyslipidemia care for diabetic patients. PMID- 16378791 TI - Effects of structured care by a pharmacist-diabetes specialist team in patients with type 2 diabetic nephropathy. AB - PURPOSE: We developed a disease management program for patients with Type 2 diabetic nephropathy and evaluated its effects on the time to onset of end-stage renal disease or all-cause death compared with usual care. METHODS: In a 2-year, prospective cohort study, we compared the clinical outcomes of patients managed by a structured care protocol (n = 80) to a group receiving usual care (n=80) in the same hospital. Patients aged < or =80 years with type 2 diabetes, serum creatinine 150-400 micromol/L, and micro- or macroalbuminuria were recruited. The structured care protocol was implemented by a pharmacist-diabetes specialist team with particular emphasis on periodic laboratory assessments, patient adherence, risk factors control, and use of renin-angiotensin system inhibitor. The primary endpoint was the composite of end-stage renal disease or all-cause death. Other endpoints were the rate of renal decline, processes-of-care measures, and control of risk factors. RESULTS: During 22.8+/-7.9 months of follow-up, the primary endpoint developed in 24 and 40 patients in the structured care and usual care groups, respectively (adjusted risk reduction, 60%, P< .001). Structured care (hazard ratio [95% confidence interval (CI)], 0.40 [0.23-0.68]), age (0.95 [0.93 0.98]), baseline systolic blood pressure (BP) (1.014 [1.003-1.026]), logarithm (base 10) of baseline serum creatinine (34 441 [2290-517915]), and macroalbuminuria (8.95 [1.22-65.38]) were independent predictors for the primary endpoint. Structured care slowed the rate of renal decline (P=.032). More intensive laboratory measurements, increased use of renin-angiotensin system inhibitor, and greater reductions in BP and low-density lipoprotein (LDL) cholesterol were reported by patients receiving structured care. CONCLUSIONS: Structured care delivered by a pharmacist-diabetes specialist team reduced the incidence of end-stage renal disease or death compared with usual care in patients with type 2 diabetic nephropathy. PMID- 16378792 TI - Bone mineral density in subjects using central nervous system-active medications. AB - PURPOSE: Decreased bone mineral density defines osteoporosis according to the World Health Organization and is an important predictor of future fractures. The use of several types of central nervous system-active drugs, including benzodiazepines, anticonvulsants, antidepressants, and opioids, have all been associated with increased risk of fracture. However, it is unclear whether such an increase in risk is related to an effect of bone mineral density or to other factors, such as increased risk of falls. We sought to examine the relationship between bone mineral density and the use of benzodiazepines, anticonvulsants, antidepressants, and opioids in a representative US population-based sample. SUBJECTS AND METHODS: We analyzed data on adults aged 17 years and older from the Third National Health and Nutrition Examination Survey (NHANES III, 1988-1994). Total femoral bone mineral density of 7114 male and 7532 female participants was measured by dual-energy x-ray absorptiometry. Multivariable linear regression models were used to quantify the relation between central nervous system medication exposure and total femoral bone mineral density. Models controlled for relevant covariates, including age, sex, and body mass index. RESULTS: In linear regression models, significantly reduced bone mineral density was found in subjects taking anticonvulsants (0.92 g/cm2; 95% confidence interval [CI]: 0.89 to 0.94) and opioids (0.92 g/cm2; 95% CI: 0.88 to 0.95) compared with nonusers (0.95 g/cm2; 95% CI: 0.95 to 0.95) after adjusting for several potential confounders. The other central nervous system-active drugs--benzodiazepines or antidepressants--were not associated with significantly reduced bone mineral density. CONCLUSION: In cross-sectional analysis of NHANES III, anticonvulsants and opioids (but not benzodiazepines or antidepressants) were associated with significantly reduced bone mineral density. These findings have implications for fracture-prevention strategies. PMID- 16378793 TI - Stroke subtypes among young patients with systemic lupus erythematosus. AB - PURPOSE: Systemic lupus erythematosus (lupus) is a systemic inflammatory disease associated with premature atherosclerosis, vasculitis, coagulopathy, and excessive incidence of stroke, especially among young patients. Little is known about subtypes of stroke in lupus. METHODS: A 20% sample of all the hospitalizations in the United States in the years 2001 and 2002 (N approximately 15 million) were analyzed to identify hospitalizations of young patients (age < or =50 years) with systemic lupus erythematosus (n=25704). Proportions of hospitalization for stroke subtypes were compared between the lupus group and the general population group. Age- and sex-adjusted odds ratios for stroke were calculated with logistic regression models. RESULTS: In the lupus group, there were 313 hospitalizations for stroke of which 206 hospitalizations had stroke as the primary diagnosis. Age- and sex-adjusted stroke risk was higher among the lupus group (odds ratio 1.5, 95% confidence interval 1.3-1.8). Patients with lupus had higher risk for all stroke subtypes except in subarachnoid hemorrhage in which a trend toward a lower risk was observed (odds ratio 0.57, 95% confidence interval 0.34-0.96). Although 12.3% (n=38) of stroke admissions in the lupus group resulted in in-hospital death, this case fatality rate was not statistically different from that for stroke in the general population group. CONCLUSIONS: Stroke is an important poor outcome in young patients with lupus. Compared with the general population, patients with lupus are more likely to be hospitalized for the risk of ischemic stroke and intracerebral hemorrhage. The risk of subarachnoid hemorrhage, however, seems to be lower in patients with lupus. PMID- 16378794 TI - Physician and patient perceptions in COPD: the COPD Resource Network Needs Assessment Survey. AB - PURPOSE: Chronic obstructive pulmonary disease (COPD), the fourth leading cause of death in the United States, has received disproportionately little attention from physicians and institutions. National data are lacking on patient and physician perceptions of and patterns of care for COPD. METHODS: Linked surveys were administered to national samples of patients with COPD, primary care physicians, and pulmonologists to evaluate perceptions of COPD severity and quality of life, attitudes about COPD, health insurance barriers to COPD care, sources of information, and knowledge about COPD diagnosis and treatment. RESULTS: Overall, 1023 patients with COPD and 1051 primary care physicians and pulmonologists responded to the surveys. Despite experiencing significant symptoms and high health care use, the majority of patients were satisfied with their care. Eighty-eight percent of physicians agreed with the statement that COPD is a "self-inflicted" disease, and more than one third were nihilistic about the treatment of patients who continued to smoke. Patients and physicians reported that insurance problems impeded access to therapies. Patients were generally uninformed about COPD; 54% of primary care physicians were aware of any COPD guidelines. Both patient and physician surveys demonstrated continued confusion about the diagnosis of COPD and treatment choices. There was frequent use of regular oral steroids despite demonstrated lack of efficacy and under-use of pulmonary rehabilitation despite proven efficacy. CONCLUSIONS: Patients with COPD have a high prevalence of activity limitations. Although most physicians believed that proper treatment can slow progression, inadequate knowledge and poor adherence to practice guidelines, together with insurance impediments, negatively impact COPD care. PMID- 16378795 TI - Orthostatic instability in a population-based study of chronic fatigue syndrome. AB - PURPOSE: Autonomic nervous system dysfunction has been suggested as involved in the pathophysiology of chronic fatigue syndrome. This population-based case control study addressed the potential association between orthostatic instability (one sign of dysautonomia) and chronic fatigue syndrome. SUBJECTS AND METHODS: Fifty-eight subjects who fulfilled criteria of the 1994 chronic fatigue syndrome research case definition and 55 healthy controls participated in a 2-day inpatient evaluation. Subjects had been identified during a 4-year population based chronic fatigue syndrome surveillance study in Wichita, Kan. The present study evaluated subjects' current medical and psychiatric status, reviewed past medical/psychiatric history and medication use, used a stand-up test to screen for orthostatic instability, and conducted a head-up tilt table test to diagnose orthostatic instability. RESULTS: No one manifested orthostatic instability in the stand-up test. The head-up tilt test elicited orthostatic instability in 30% of eligible chronic fatigue syndrome subjects (all with postural orthostatic tachycardia) and 48% of controls (50% with neurally mediated hypotension); intolerance was present in only nonfatigued (n=7) subjects. Neither fatigue nor illness severity were associated with outcome. CONCLUSIONS: Orthostatic instability was similar in persons with chronic fatigue syndrome and nonfatigued controls subjects recruited from the general Wichita population. Delayed responses to head-up tilt tests were common and may reflect hydration status. These findings suggest reappraisal of primary dysautonomia as a factor in the pathogenesis of chronic fatigue syndrome. PMID- 16378796 TI - The natural history of post-traumatic hypopituitarism: implications for assessment and treatment. AB - PURPOSE: Hypopituitarism has been reported in up to half of long-term survivors of traumatic brain injury. We attempted to define the natural history of post traumatic hypopituitarism to devise guidelines for the optimal timing of patients' assessment and hormone replacement. SUBJECTS AND METHODS: Fifty consecutive patients with severe or moderate head trauma were enrolled in a prospective study of pituitary function during the acute phase, at 6 months, and at 12 months after injury. Growth hormone and adrenocorticotropin hormone reserves were assessed using the glucagon stimulation test. Baseline serum concentrations of other anterior pituitary hormones were measured. Results were compared with normative data obtained from matched healthy controls. RESULTS: Nine patients (18%) had growth hormone deficiency in the acute phase; at 6 months, 5 patients recovered function and 2 new deficiencies were detected; at 12 months, 1 patient recovered, leaving 5 patients (10%) with growth hormone deficiency. Eight patients (16%) showed subnormal cortisol response in the acute phase; at 6 months, 4 patients had recovered and 5 new deficiencies were detected; all 9 patients had persistent abnormalities at 2 months. Forty patients (80%) had gonadotropin deficiency in the acute phase, of whom 29 (73%) recovered by 6 months and 34 (85%) recovered by 12 months. Thyrotropin deficiency was present in 1 patient in the acute phase, who recovered by 6 months; 1 new case was diagnosed at 6 months, which persisted at 12 months. CONCLUSION: After traumatic brain injury, early neuroendocrine abnormalities are sometimes transient, whereas late abnormalities present during the course of rehabilitation. A follow-up strategy with periodic evaluation is a necessary part of the optimal care for patients with traumatic brain injury. PMID- 16378797 TI - Staphylococcus aureus bacteremia in patients with prosthetic devices: costs and outcomes. AB - PURPOSE: Although Staphylococcus aureus is a leading cause of nosocomial infection, little is known about the impact of S. aureus bacteremia on patients with prosthetic devices. This investigation sought to define the clinical outcome, health care resource use, and infection-associated costs of S. aureus bacteremia in patients with prostheses. SUBJECTS AND METHODS: All hospitalized patients with a prosthetic device and S. aureus bacteremia during the 96-month study period were identified prospectively. Clinical data were collected at the time of hospitalization. Data regarding infection-related resource utilization and infection-related costs within 12 weeks of the initial bacteremia were also recorded. RESULTS: 298 patients with > or =1 prosthesis and S. aureus bacteremia were identified (cardiovascular device--122 patients, orthopedic device--73 patients, long-term catheter--71 patients, and other devices-32 patients). Overall, 58% of patients underwent surgery as a consequence of the infection. Infection-related complications occurred in 41% and the overall 12-week mortality was 27%. The mean infection-related cost was 67439 dollars for patients with hospital-acquired S. aureus bacteremia and 37868 dollars for community-acquired S. aureus bacteremia (cost difference 29571 dollars; 95% confidence interval, 14370 dollars-49826 dollars). Rates of device infection, complications, 12-week mortality, and mean cost varied by prosthesis type. CONCLUSION: S. aureus bacteremia in patients with prosthetic devices is associated with frequent complications, substantial cost, and significant health care resource utilization. PMID- 16378798 TI - Cystatin-C and inflammatory markers in the ambulatory elderly. AB - PURPOSE: Inflammatory factors are elevated in persons with severe renal dysfunction, but their association across all levels of renal function is unclear. We compared cystatin-C, a novel marker of renal function, with creatinine and estimated glomerular filtration rate (eGFR) as predictors of C reactive protein and fibrinogen levels. METHODS: This study is a cross-sectional analysis to evaluate cystatin-C, creatinine, and eGFR as predictors of the inflammatory markers C-reactive protein and fibrinogen. Participants included 4637 ambulatory elderly patients from the Cardiovascular Health Study. Multivariate linear regression was used to determine the independent associations of each renal function measurement with the inflammatory marker outcomes. RESULTS: After adjustment for confounding factors, cystatin-C was correlated with both C-reactive protein (coefficient = 0.13; 95% confidence interval: 0.10-1.16, P <.0001) and fibrinogen levels (0.15; 0.13-0.18, P <.0001). Associations were larger than those for creatinine and C-reactive protein (0.05; 0.02-0.07, P = .003) or fibrinogen (0.07; 0.04-0.10, P <.0001). Adjusted levels of C-reactive protein increased incrementally across quintiles of cystatin-C, from a median of 2.2 mg/L in quintile 1 to 3.7 mg/L in quintile 5. In contrast, both C-reactive protein and fibrinogen had U-shaped associations with quintiles of creatinine and eGFR, because the inflammatory markers were equivalently elevated in quintiles 1 and 5. CONCLUSIONS: The finding of a significant linear association of cystatin-C and inflammation markers suggests that even small reductions in renal function may be associated with adverse pathophysiologic consequences. PMID- 16378799 TI - Herpes zoster in immunocompromised patients: incidence, timing, and risk factors. AB - PURPOSE: To evaluate the risk factors for herpes zoster as well as the incidence and timing of this complication in patients who were treated with immunosuppression because of active Wegener's granulomatosis. SUBJECTS AND METHODS: We studied the 180 Wegener's granulomatosis patients in the Wegener's Granulomatosis Etanercept Trial (WGET). Herpes zoster events during WGET were documented prospectively. Follow-up questionnaires were employed to describe the location, treatment, and complication(s) of herpes zoster and its therapy. Univariate and multivariate analyses were performed to evaluate risk factors, including history of herpes zoster, for the occurrence of herpes zoster during the trial. All analyses were based on the time to first occurrence of herpes zoster. RESULTS: Eighteen patients (10% of the WGET cohort) suffered a total of 19 herpes zoster episodes over a mean follow-up period of 27 months. The annual incidence of herpes zoster in the WGET cohort was 45 cases/1000 patient-years (95% confidence interval [CI]: 27, 70). The median time from enrollment to the occurrence of herpes zoster in the subgroup of patients with that complication was 16.5 months (+/- 9.4). Fifteen of the 19 herpes zoster events (79%) occurred between months 6 and 36, many months after the period of most intensive immunosuppression. In univariate analyses, history of serum creatinine > or =1.5 mg/dL before enrollment was associated with a relative risk (RR) of 3.0 (95% CI: 1.1, 7.8) for herpes zoster during WGET (P=.03). In multivariate analyses, serum creatinine > or =1.5 mg/dL was associated with an RR of 6.3 (95% CI: 2.0, 19.8; P=.002), and female sex with an RR of 4.6 (95% CI: 1.6, 13.2; P=.004). CONCLUSION: Renal dysfunction and female sex were consistently strong risk factors for herpes zoster events in this population. Contrary to expectation, most herpes zoster events did not occur during periods of most intensive immunosuppression. These data may inform studies of interventions designed to prevent herpes zoster in patients on treatment for immune-mediated diseases. PMID- 16378800 TI - The degree of chills for risk of bacteremia in acute febrile illness. AB - PURPOSE: Patients with acute febrile illness may experience different degrees of chills. To evaluate the different degrees of chills in predicting risk of bacteremia in patients with acute febrile illness, we performed a single-center prospective observational study. METHODS: We enrolled consecutive adult patients with acute febrile illness presenting to our emergency department. We defined mild chills as cold-feeling equivalent such as the need of an outer jacket; moderate chills as the need for a thick blanket; and shaking chills as whole-body shaking even under a thick blanket. We estimated risk ratios of the different degrees of chills for bacteremia using multivariable adjusted Poisson regression. RESULTS: Of a total 526 patients, 40 patients (7.6%) had bacteremia. There were 65 patients (12.4%) with shaking chills, 100 (19%) with moderate chills, and 105 (20%) with mild chills. By comparing patients with no chills, the risk ratios of bacteremia were 12.1 (95% confidence interval [CI] 4.1-36.2) for shaking chills, 4.1 (95% CI 1.6-10.7) for moderate chills, and 1.8 (95% CI 0.9-3.3) for mild chills. Shaking chills showed a specificity of 90.3% (95% CI 89.2-91.5) and positive likelihood ratio of 4.65 (95% CI 2.95-6.86). The absence of chills showed a sensitivity of 87.5% (95% CI 74.4-94.5) and negative likelihood ratio of 0.24 (95% CI 0.11-0.51). CONCLUSION: Evaluation of the degree of chills is important for estimating risk of bacteremia in patients with acute febrile illness. The more severe degree of chills suggests the higher risk of bacteremia. PMID- 16378801 TI - D-dimer is not useful for the diagnosis of isolated superficial venous thrombosis. PMID- 16378802 TI - The safety of levofloxacin in elderly patients on warfarin. PMID- 16378803 TI - Safety of calcium antagonists: more ACTION. PMID- 16378804 TI - Characterizing Gulf War Illnesses: neurally mediated hypotension and postural tachycardia syndrome. PMID- 16378805 TI - Risk of first-time hospitalization for angioedema among users of ACE inhibitors and angiotensin receptor antagonists. PMID- 16378806 TI - Restructuring the academic department of internal medicine. PMID- 16378807 TI - Hepatocellular carcinoma with extension into the right atrium. PMID- 16378808 TI - Very high cerebrospinal fluid protein in a patient with hyperosmolar coma. PMID- 16378809 TI - Azithromycin-induced hepatoxicity. PMID- 16378810 TI - Amelioration of differentiation of dendritic cells from CVID patients by intravenous immunoglobulin. PMID- 16378812 TI - A 57-year-old man with a 6-day headache and fatigue. PMID- 16378813 TI - Malaria or typhoid fever: a diagnostic dilemma? PMID- 16378815 TI - Primary care-based disease management for patients with diabetes. PMID- 16378817 TI - Improving the management of terminal heart failure patients. PMID- 16378818 TI - Management of heart failure in the elderly. PMID- 16378820 TI - Self-monitoring of blood glucose cannot compensate for ineffective diabetes management. PMID- 16378822 TI - Initiation of warfarin therapy in elderly medical patients: a safe and accurate regimen. PMID- 16378824 TI - Subglottic suctioning to prevent ventilator-associated pneumonia. PMID- 16378826 TI - An analysis of the published Massachusetts General Hospital case records (1994 2004). PMID- 16378827 TI - There is more in the ECG than is reported. PMID- 16378828 TI - Light of my life. PMID- 16378829 TI - Transient endothelial dysfunction is present shortly after cardioversion in patients with lone atrial fibrillation. PMID- 16378830 TI - Interobserver agreement on ultrasound measurements of residual vein diameter, thrombus echogenicity and Doppler venous flow in patients with previous venous thrombosis. AB - INTRODUCTION: In patients with new symptoms in a leg previously affected by deep vein thrombosis (DVT), the presence of thrombus on ultrasound cannot be assumed to be due to recurrent thrombosis. Several parameters have been suggested to differentiate between acute and chronic thrombus on ultrasound, including measurement of residual vein diameter during compression, thrombus echogenicity and Doppler assessment of venous flow, but studies on the reproducibility of these measurements are sparse. OBJECTIVE: To determine interobserver agreement on measurement of residual vein diameter, thrombus echogenicity and Doppler venous flow in patients with residual thrombus in the veins of the lower limb. MATERIALS AND METHODS: Patients with previous proximal DVT who had a high likelihood of residual thrombosis, but without symptoms of recurrent DVT, had ultrasound examinations independently performed by two examiners on the same day. Interobserver agreement on measurement of residual vein diameter, thrombus echogenicity and Doppler venous flow was evaluated. RESULTS: We determined that interobserver agreement on these measurements was moderate. The mean difference between paired measurements of residual vein diameter was 2.2 mm (95th centile, 8.0 mm). When both examiners agreed residual thrombus was present, 54% of the variance of the measurement of residual vein diameter was accounted for by the paired measurements. The weighted kappa coefficient for thrombus echogenicity was 0.01 and for Doppler venous flow was 0.51. CONCLUSIONS: The error associated with ultrasound measurements of residual vein diameter, thrombus echogenicity and flow appears to be considerable. PMID- 16378831 TI - Comparison of platelet fibronectin, ADP-induced platelet aggregation and serum total nitric oxide (NOx) levels in angiographically determined coronary artery disease. AB - INTRODUCTION: Coronary thrombosis is an important determinant of prognosis in patients with acute coronary syndromes. Fibronectin is also found in platelets within the alpha secretory granules and secreted following platelet stimulation by a variety of agonist. Available data suggest that expression of platelet fibronectin on the cell surface may indicate a role in platelet aggregation and adhesion to fibrin thrombi and connective tissue. Clear evidence has emerged that a concerted action of nitric oxide (NO) generated by either endothelial or platelet NO synthases regulates platelet activation, causing inhibition of adhesion and aggregation. The aim of the present study was determining and correlating the serum total NO (NOx), platelet fibronectin and ADP-induced platelet aggregation levels in coronary artery disease (CAD) patient subgroups. MATERIALS AND METHODS: A total of 43 coronary artery disease patients were included in this study. Peripheral blood samples from patients with coronary artery disease were obtained from the Department of Cardiology. Platelet aggregation tests with adenosine diphosphate (ADP) were analyzed by using aggregometer. Platelet fibronectin concentrations were determined by enzyme linked immunosorbent assay (ELISA). Serum total nitric oxide (NOx) levels were determined by colorimetric method. RESULTS: In patients with double-vessel disease, platelet fibronectin levels were found to be significantly higher than no-vessel disease (p<0.001), single-vessel disease (p<0.01) and triple-vessel disease (p<0.001). In addition, in patients with single-vessel disease platelet fibronectin levels significantly higher than no-vessel disease (p<0.05). We could not find any significant differences in ADP-induced platelet aggregation and serum NOx values between CAD patient subgroups. There was a positive correlation between platelet fibronectin levels and severity of disease (r=0.315, p<0.05). PMID- 16378832 TI - Acute idiopathic portal vein thrombosis in a child: a case report and literature review. PMID- 16378833 TI - Increase in platelet non-integrin type I collagen receptor in patients with systemic sclerosis. AB - Microvascular injury is one of the major pathogenetic processes involved in systemic sclerosis (SSc). Interaction of the platelet types I and III collagen receptors with their respective ligand in the exposed subendothelial stroma as a result of ongoing microvascular injury in SSc patients results in platelet activation and aggregation with the release of mediators, which contribute to vascular damage and inflammation. We have found that there is a twofold increase in radiolabeled type I collagen binding to washed platelets from patients with SSc compared to platelets obtained from normal volunteers. Western blot analyses showed that the non-integrin platelet type I collagen receptor protein (65 kDa) is increased dramatically in lysates of platelet from patients with SSc. However, the integrin (alpha(2)beta(1)) and other non-integrin receptors such as glycoprotein VI, glycoprotein IV, and the platelet receptor for type III collagen remain unchanged. In addition, platelet lysates from rheumatic disease controls (rheumatoid arthritis, osteoarthritis, gout, and systemic lupus erythematosus) do not show any significant increases. There is no nitrotyrosylation on 65 kDa in patients with SSc compared to controls, suggesting this might also contribute to binding of CI to the 65-kDa CIR. These results suggest that there is a specific increase in the number of platelet type I collagen receptors in SSc patient's platelets. In addition, the activity of nitric oxide synthase is decreased in patients' platelet lysates compared to controls. The increase in platelet expression of the 65-kDa non-integrin platelet type I collagen receptor may explain the enhanced aggregation of platelets from patients with SSc to CI in vitro and microvascular thrombosis in the disease in vivo. PMID- 16378834 TI - Structures of importance for the stability of antiplasmin as studied by site directed mutagenesis. AB - Human antiplasmin, a fast-acting inhibitor of plasmin in plasma, belongs to the serpin super-family of proteins. Like other members of this family, antiplasmin has a scissile peptide bond exposed within a reactive centre loop, typically present at the surface of the molecule. Antiplasmin is stable at neutral pH, but at acidic pH or at elevated temperatures it rapidly becomes inactivated. Data regarding "native" antiplasmin have demonstrated that both polymerization processes and formation of latent molecules are important in this respect. In this work we used site-directed mutagenesis to produce 11 single-site mutants (mainly within Abeta-sheet, Bbeta-sheet and reactive centre loop), which were expressed in Drosophila S2 cells, purified and characterized. Five of the 11 mutants were found to have a deviating stability at decreased pH. Glu346Thr was the only mutant with a lesser stability as compared to wt-antiplasmin, but the other 4 were more stable. The most stable mutant, His341Thr, was 7-fold more stable at pH 4.9 as compared to wt-antiplasmin. The wt-antiplasmin had a much more pronounced tendency to polymerize at decreased pH, as compared to "native" antiplasmin. However, many of the mutants clearly rather formed latent molecules, as judged both from PAGE-analysis at non-denaturing condition and reactivation experiments. PMID- 16378835 TI - The antithrombotic and anti-inflammatory effects of BCX-3607, a small molecule tissue factor/factor VIIa inhibitor. AB - Tissue factor (TF) is a transmembrane glycoprotein that binds its zymogen cofactor, Factor VIIa (FVIIa) on the cell surface. Together (TF/FVIIa) they activate Factor X (FX) and Factor IX (FIX) and start the extrinsic pathway of blood coagulation. As such, the TF/FVIIa complex plays an important role in normal physiology as well as in thrombotic diseases such as unstable angina (UA), disseminated intravascular coagulation (DIC), and deep vein thrombosis (DVT). In addition to its function as an initiator of coagulation, TF/FVIIa plays an important role in inflammation. Expression of TF on the cell surface and its appearance as a soluble molecule are characteristic features of acute and chronic inflammation in conditions such as sepsis and atherosclerosis. Here we demonstrate that BCX-3607, a small molecule potent inhibitor of TF/FVIIa, reduces thrombus weight in an animal model of DVT. BCX-3607 also decreases the level of interleukin-6 (IL-6) in a LPS-stimulated mouse model of endotoxemia. Additionally, in vitro studies indicate that BCX-3607 blocks the generation of TF/FVIIa-induced IL-8 mRNA in human keratinocytes and reduces the TF/FVIIa mediated generation of IL-6 and IL-8 in human umbilical vein endothelial cells (HUVEC). Therefore, BCX-3607 might block the TF/FVIIa-mediated coagulation and inflammation associated with pathological conditions. PMID- 16378837 TI - Branched stents--endovascular evolution continues. Endoscopic surgery of the brain, skull base, and spine. Statins for vasospasm. PMID- 16378838 TI - Failed back surgery patients: the alarm bells are ringing. PMID- 16378839 TI - Lack of association between the IL1A gene (-889) polymorphism and outcome after head injury. AB - BACKGROUND: Interleukin (IL) 1 is a proinflammatory cytokine that has been identified as an important mediator of neurodegeneration induced by ischemia or traumatic brain injury. Accumulating evidence to date has suggested that the major cytokine contributing to neurodegeneration after head injury is IL-1beta rather than IL-1alpha; however, there is no sufficient data regarding IL-1alpha in literature, and there may be an association between IL1A gene polymorphism and outcome after head injury. METHODS: We performed a prospective clinical study and included a recruited series of 71 patients who had head injury and were admitted to our neurosurgical unit. Severity of initial injury was assessed by the Glasgow Coma Scale. Outcome at 6 months after injury was assessed by means of the Glasgow Outcome Score. Interleukin 1alpha genotypes were determined from blood samples by standard methods. RESULTS: Of 40 patients with IL1A*2, 18 (45%) had an unfavorable outcome (dead, vegetative state, or severe disability) compared with 7 (22.5%) of 31 without IL1A*2 (P = .08). CONCLUSION: Our findings show that there is no genetic association between IL1A gene polymorphism and outcome after head injury. Further clinical studies should be designed to confirm and further evaluate these findings. PMID- 16378841 TI - Intracranial hemangiopericytomas: correlation of topoisomerase IIalpha expression with biologic behavior. AB - BACKGROUND: Meningeal hemangiopericytomas are aggressive tumors that have a high rate of recurrence despite gross total resection and radiation therapy. Topoisomerase, a cell proliferation marker, is also a target of certain chemotherapeutic agents, and its nuclear levels are speculated to predict efficacy of targeted therapy. The aim of this study was to correlate the topoisomerase IIalpha proliferation index (TPI) with biologic behavior in intracranial hemangiopericytomas. METHODS: Clinical, radiological, and management data in 27 patients with intracranial hemangiopericytoma admitted between 1990 and 2003 were reviewed. Immunohistochemistry was performed on all the tumors using a monoclonal antibody to topoisomerase IIalpha, and the proliferation index was calculated. The effect of TPI on outcome was sought. RESULTS: The male/female ratio was 15:12. The mean age at presentation was 31.33 years. A radical excision of tumor was done in 18, subtotal excision in 2, partial excision in 4, and a biopsy in 3 patients. Tumor recurrence was noted in 15 (55.6%) of the 27 patients (mean follow-up duration, 51.5 months). The time to recurrence ranged from 7 months to 8 years (mean, 49 months). The 5-year recurrence-free survival was 33.8% in patients with a TPI of 5% or greater, and 72% in patients with a TPI of less than 5%. The relative risk of recurrence was 2.9 times greater in patients with a TPI 5% or greater as compared with those a TPI of less than 5%. CONCLUSION: Our study suggests that cases with a radical excision, radiation therapy, or a TPI index of less than 5% have a longer recurrence-free survival. A TPI of 5% or greater is a reliable predictor of recurrence. PMID- 16378842 TI - Size and location of ruptured and unruptured intracranial aneurysms measured by 3 dimensional rotational angiography. AB - OBJECTIVE: The aim of the study was to report about accurate size and location of a consecutive series of ruptured and unruptured aneurysms taking the complex 3 dimensional (3D) anatomy and parent vessel morphology into consideration by using the newly developed 3D rotational angiography (3D-RA). METHODS: One hundred eighteen consecutive patients with 155 saccular intracranial aneurysms were included in the study and received 3D-RA reconstructions for measurement of maximal height and width of the aneurysmal sac. Statistical evaluation compared values for ruptured (n = 83) and unruptured (n = 72) aneurysms. RESULTS: Mean height and width of unruptured aneurysms were 5.7 and 5.7 mm; of ruptured aneurysms, 6.7 and 6.1 mm (not significant, P = .7 for height and P = .9 for width). The majority of ruptured aneurysms, 81.9% and 59%, were smaller than 10 and 7 mm; likewise, 81.9% and 68.1% of unruptured aneurysms were smaller than 10 and 7 mm. The difference in frequency of small (<10/<7 mm) aneurysms between unruptured and ruptured aneurysms was not significant (P = 1.0 and .32, respectively). The majority (69.4%) of small ruptured aneurysms (<7 mm) were located in the anterior circulation. Most ruptured aneurysms were in the size group 4 to 6 mm in height and 2 to 4 mm in width, and a critical threshold size for aneurysm rupture could not be identified. CONCLUSIONS: An automated calibration procedure applied to all images and excellent visualization of aneurysm and parent vessel morphology using 3D-RA allow accurate size measurement of intracranial aneurysms which may be smaller than previously thought. Small aneurysm (<7 mm), also in the anterior circulation, should be carefully evaluated for treatment. PMID- 16378847 TI - Fractionated stereotactic radiotherapy in the treatment of exclusive cavernous sinus meningioma: functional outcome, local control, and tolerance. AB - BACKGROUND: Fractionated stereotactic radiotherapy (FSRT) combines the precision of stereotactic positioning with the radiobiologic advantage of dose fractionation. METHODS: From June 1997 to June 2001, 30 patients with cavernous sinus meningiomas were treated with FSRT using fixed noncoplanar conformal fields. Patient skull fixation was achieved using the BrainLAB mask (20 patients) or Beverly frame (10 patients). The Cosman-Roberts-Wells coordinate frame was used for stereotactic space definition. In selected cases before 1999, and in all cases afterward, gadolinium-enhanced MRI for image fusion was performed. The median radiation dose was 52 Gy, with a daily fraction of 2 Gy. Patients were regularly followed up analyzing symptoms, tumor progression, and side effects. Neurocognitive function was evaluated retrospectively for 26 patients using Mini Mental State Examination. RESULTS: Median follow-up period was 50 months (range, 28.2-74.5 months). Preexisting neurologic symptoms improved in 50% of the patients and worsened in 2 patients. Only 2 patients progressed and the actuarial local progression free survival was 93% at 4 years. Tolerance was good with 2 cases of late radiation toxicity which consisted of moderate short-term memory loss and dysphasia in one case and neuropsychologic deficit with seizures in the other. Postradiotherapy Mini-Mental State Examination results showed a median score of 28 (range, 16-30). CONCLUSIONS: Fractionated stereotactic radiotherapy is a high-precision technique. It is safe and feasible in the primary and adjuvant treatment of cavernous sinus meningiomas. Fractionated stereotactic radiotherapy allowed local control in more than 90% of patients. PMID- 16378850 TI - Advice for a young neurosurgeon. PMID- 16378851 TI - Water dissection technique of Toth for opening neurosurgical cleavage planes. AB - BACKGROUND: The low-pressure water dissection technique of Toth, first reported in 1987, is a method to cautiously open neurosurgical cleavage planes such as the sylvian fissure or the interhemispheric space, and the interfaces between extraparenchymal masses and the adjacent brain. The aim of this technical report is to present our long-term experience with this simple and elegant asset of microneurosurgery and to promote its widespread use. METHOD: Water is injected under microscopic control by a hand-held syringe with a blunt needle or by an irrigating balloon applying repeated injections of physiological saline into the cleavage plane to open it. FINDINGS AND CONCLUSION: The water dissection technique of Toth has been extensively used in Budapest and Helsinki in thousands of microsurgical cases, in removal of meningiomas and to open sylvian and interhemispheric fissure. In our experience, there have been no noticeable complications, and we recommend this technique for widespread use. It is a very inexpensive, simple, and effective method not requiring any expensive or complicated devices. PMID- 16378853 TI - Comparative dural closure techniques: a safety study in rats. AB - BACKGROUND: Some neurosurgical procedures have high morbidity and mortality rates due to cerebrospinal fluid (CSF) fistula development, particularly when dural defects are in relatively inaccessible areas or surrounded by friable dura. We used a rat model to test 4 different dural closure techniques to determine which one was significantly superior for achieving a watertight dural closure with minimal harm to brain tissue. METHODS: The rats were randomly divided into 2 groups. The first group (group A, n = 40) was used to test the strength of the adhesivity for CSF leakage. Histopathologic studies were used to evaluate the granulation tissue between the dura and dural graft. Effects on the brain tissue were studied in the second group (group B, n = 40) where lipid peroxidation was determined. These 2 groups consisted of 5 subgroups: control, methyl metacrylate, n-butyl cyanoacrylate, fibrin glue, and CO(2) laser. RESULTS: Methyl metacrylate and CO(2) laser techniques were inadequate for stopping dural leakage and had harmful effects on brain tissue. Cerebrospinal fluid leak was observed only in 1 rat in the n-butyl cyanoacrylate subgroup and this result was statistically significant (P = .0005), but lipid peroxidation levels for this material showed that it was not safe for dural closure in case it leaked through the dural defect. The lipid peroxidation levels of the fibrin glue subgroup were not statistically significantly different from the control group (P = .440). CONCLUSIONS: Fibrin glue was the safest material with a CSF leakage risk that was higher than n-butyl cyanoacrylate (25% vs 12.5%) but acceptable. This study showed no relationship between the CSF leak and histopathologic findings for sealant properties of the tissue adhesives. PMID- 16378855 TI - The use of a sump antibiotic irrigation system to save infected hardware in a patient with a vagal nerve stimulator: technical note. AB - The authors describe the use of a sump irrigation system that was used to successfully treat the battery implantation site of a vagal nerve stimulator (VNS). Irrigation was composed of a dilution of vancomycin in lactated Ringer's solution. At long-term follow up, the patient has not returned with signs or symptoms of wound infection. She continues to effectively combat her epilepsy with VNS. The authors believe this to be the first description of this technique for salvaging an implanted VNS. PMID- 16378858 TI - Fronto-orbital craniotomy reconstruction using the titanium clamp (Craniofix): a technical note. AB - BACKGROUND: We evaluate a new technique for plateless fixation of a bone flap after fronto-orbital craniotomy. METHODS: From September 1999 to October 2004, we performed fronto-orbital craniotomy reconstruction using the Craniofix titanium clamp in 108 consecutive patients with a variety of lesions in the anterior skull base. Postoperative computed tomographic imaging studies and clinical evaluations were performed to prospectively assess cosmetic conformity and bone flap stability and to evaluate the surgical benefit of Craniofix in these patients. RESULTS: Excellent bone flap fixation and cosmetic results were obtained in all patients 6 to 68 months (average, 36 months) after surgery. CONCLUSION: The Craniofix titanium clamp is a reliable, safe, and simple fixation device for reconstruction of fronto-orbital craniotomy. PMID- 16378859 TI - Spontaneous arteriovenous fistula of the orbit: case report. AB - BACKGROUND: A spontaneous arteriovenous fistula of the orbit is exceedingly rare. We report the first case of such condition treated with direct surgical exposure of the superior ophthalmic vein (SOV) followed by embolization. CASE DESCRIPTION: A 55-year-old man presented with upper lid swelling, conjunctival chemosis, and proptosis of the right eye. Angiography revealed an intraorbital shunt supplied by the ophthalmic artery whose venous drainage curved anteriorly to the SOV. Because transarterial and transvenous endovascular approaches to treat the fistula were impossible, we performed direct surgical exposure of the SOV followed by the embolization of the fistula. Postoperative angiograms demonstrated complete closure of the fistula. All symptoms had disappeared by 2 months after surgery. CONCLUSIONS: For appropriate treatment planning, it is necessary to identify the location of the shunt. In cases where transarterial and transvenous endovascular approaches to treat the fistula are difficult or impossible, direct surgical exposure of the SOV followed by embolization may accomplish complete closure of the fistula without significant risk for iatrogenic injury. PMID- 16378861 TI - Acute combination fracture of atlas and axis: "triple" anterior screw fixation in a 92-year-old man: technical note. AB - BACKGROUND: The combination of odontoid and bilateral transarticular C1-C2 anterior screw fixation is a recent addition in treating C1-type II odontoid fractures. When feasible, it ensures early maximal stability, even if it slightly reduces the mobility of C1-C2 complex. We report a case of combination atlas-type II odontoid fracture that occurred in a 92-year-old man. The instability was treated with odontoid screw fixation and anterior bilateral C1-C2 transarticular screw fixation in a single stage. The aim of the article is to describe the feasibility of "triple" anterior screw fixation in the presence of C1-type II odontoid fracture. METHODS: The diagnosis, treatment, and outcome of a 92-year old patient with mild tetraparesis caused by C1-type II odontoid fracture were assessed. RESULTS: Cervical x-rays, computed tomographic scan, and magnetic resonance imaging demonstrated a fracture of posterior arch of C1, associated with type II odontoid fracture and with presumable damage of C1 transverse ligament. Magnetic resonance imaging also showed a high cervical centromedullary area slightly hyperintense in T1-weighted images. Treatment consisted of odontoid and bilateral C1-C2 transarticular screw fixation with single anterior approach. The admission neurologic conditions improved and the patient was early mobilized. CONCLUSIONS: The authors suggest that in presence of C1-type II odontoid fracture, the triple anterior screw fixation has to be taken into account as salvage procedure, especially if other methods of stabilization failed or cannot be safely performed. This technique seems to be safety feasible also in old patients, as our report and the experience of others confirm. PMID- 16378862 TI - Simple and safe method of cranial reconstruction after posterior fossa craniectomy. AB - BACKGROUND: Reconstructing the posterior fossa after surgical intervention in this region is important both for prevention of postoperative complication such as headache and for cosmetic purposes. Several methods have been reported that use either synthetic or natural graft, the latter being either autograft or allograft. The previously described methods require either surgical intervention on a second setting or an additional procedure that prolongs the time of the surgery and may contribute to morbidity. The present report describes a simple modified method of reconstructing the postcraniectomy defect by using the patients' own bone dust, tissue glue, and gel foam sheets. METHODS: The method of reconstruction requires collection of as much as possible of the produced bone dust at the time of craniectomy, adding tissue glue, placing in between 2 sheets of gel foam, and shaping it to match the surgical defect. RESULTS: Ten cases of various posterior fossa pathologies managed at King Faisal hospital of the university between January 2000 and September 2004 had reconstruction of the posterior fossa after craniectomy during the same operative setting using the described method of cranial reconstruction. No complication was noted. Patients did not have any delayed postcraniectomy pain at reconstruction site. Postoperative plane x-ray of the skull and computed tomography showed good healing and shaping of the suboccipital bone at the surgical defect. CONCLUSIONS: Reconstructive cranioplasty is an important part of any posterior fossa exposure. The present report describes a safe and simple method that gives acceptable results both clinically and radiologically. PMID- 16378863 TI - Surgery for spontaneous intracerebral hemorrhage has greater remedial value than conservative therapy. AB - BACKGROUND: The aim of this study was to compare the efficacy of surgery for spontaneous intracerebral hemorrhage with that of medical treatment, based on data from the Japan Stroke Registry Study. METHODS: From 1999 to 2001, 1010 patients with spontaneous intracerebral hemorrhage were registered in the Japan Standard Stroke Registry Study from 45 stroke center hospitals in Japan. The National Institutes of Health Stroke Scale (NIHSS), Japan Stroke Scale (JSS), and modified Rankin Scale scores were used to compare severity and improvement in patients given surgical and medical treatment. CONCLUSIONS: Surgically treated patients, especially those with cerebellar hemorrhage, had significantly greater improvement in NIHSS or JSS score compared with medically treated patients. Our findings indicated that the patients who underwent surgery appeared to have better outcomes. But, because the study was not randomized, this observation cannot be interpreted as indicating that surgery is advantageous. PMID- 16378864 TI - A long-term ventricular drainage for patients with germ cell tumors or medulloblastoma. AB - BACKGROUND: Hydrocephalus associated with intracranial germ cell tumors or disseminated medulloblastoma has been treated with ventriculoperitoneal shunt. However, this procedure has a potential risk of intraperitoneal metastasis of these brain tumors. To prevent this potential risk and to minimize the risk of infection, we developed a percutaneous long-tunneled ventricular drainage (PLTVD). To confirm the effectiveness, we retrospectively analyzed the results of this procedure. METHODS: From 1979 to 2003, we have treated 96 patients with germ cell tumors and medulloblastoma in our hospital. Of 96 patients, 59 (germ cell tumor, 31; medulloblastoma, 28) had hydrocephalus and 13 needed long-term cerebrospinal fluid drainage to manage the obstructive hydrocephalus due to persistent tumor or communicating hydrocephalus due to dissemination. We performed PLTVD for these cases using a flow-controlled shunt device and percutaneous long-tunneled shunt tube (peritoneal catheter) exiting at the upper abdomen and connecting to a closed drainage system. The occurrence of extraneural metastasis and the incidence of infection were evaluated. RESULTS: The average duration of drainage was 74 days (range, 34-115 days). All 13 cases received full dose chemotherapy and radiotherapy without infectious complications or extraneural metastasis. CONCLUSIONS: Percutaneous long-tunneled ventricular drainage was an effective method to manage long-lasting obstructive or communicating hydrocephalus with germ cell tumors and medulloblastoma. PMID- 16378868 TI - Intracranial metastatic parathyroid carcinoma: case report. AB - BACKGROUND: Although parathyroid carcinoma is not frequent, it is a slowly progressive disease characterized by frequent recurrences. A review of the literature revealed only 2 other cases of intracranial metastatic parathyroid carcinoma. We present here the case of cerebral metastases from parathyroid carcinoma that could be treated successfully. CASE DESCRIPTION: This 61-year-old Japanese woman presented to our institute with a complaint of right lower extremity weakness after hemiconvulsion. She had undergone a parathyroidectomy for parathyroid carcinoma 18 years earlier. Lung metastasis was also detected 6 years earlier, and she has been dialyzed twice a month after chemotherapy. Computed tomographic scans demonstrated 2 enhancing right frontal tumors. After resection of the intracranial metastases, her right hemiparesis and secondary hyperparathyroidism resolved. CONCLUSIONS: This case report supports aggressive surgical management to eliminate all parathyroid hormone-secreting malignant tissue and prevent metabolic complications. PMID- 16378869 TI - Hemorrhagic colloid cyst: case report and review of the literature. AB - Colloid cysts are benign tumors situated in and connected to the roof of the third ventricle. The clinical presentation is diverse, varying from incidentally found cysts to acute death. This presentation is explained by an acute hydrocephalus due to an obstruction of the cerebrospinal fluid (CSF) flow at the level of the foramina of Monro. Why these lesions cause a sudden obstruction is not clear in most cases. Possible explanations of a sudden deterioration are shifts of the cysts after lumbar puncture or CSF flow obstruction after shunt dysfunction. We add an explanation for the acute deterioration of patients with colloid cysts (ie, hemorrhagic changes in the cysts). This complication has been published only 4 times before, all diagnosed at postmortem examination. A young patient is presented with a recently diagnosed and symptomatic colloid cyst who deteriorated the night before surgery because of intracystic hemorrhage. Despite emergent CSF diversion, our patient remained in a poor clinical condition. We review the literature about this topic and discuss the consequences for the neurosurgical practice. PMID- 16378870 TI - Pneumocephalus in a patient with a ventriculoperitoneal shunt after percutaneous gastrojejunostomy catheter placement: case report. AB - BACKGROUND: Percutaneous gastrostomy and/or jejunostomy associated with ventriculoperitoneal (VP) shunting in critically ill neurosurgical patients is not an uncommon combination. Massive intraventricular pneumocephalus has not been previously reported as a complication of percutaneous gastrostomy and/or jejunostomy placement in a patient with a VP shunt. A case is presented here where we believe such a complication occurred. CASE DESCRIPTION: Our patient is a 68-year-old woman who experienced a subarachnoid hemorrhage from a right anterior choroidal aneurysm rupture. The patient underwent endovascular coiling. The patient developed a communicating hydrocephalus and eventually necessitated a VP shunt. Two weeks after shunt placement, our patient had a fluoroscopic percutaneous gastrostomy and/or jejunostomy catheter placed. A computed tomographic scan of the brain obtained after feeding tube placement for a change in mental status revealed a significant amount of air in the lateral ventricles. The patient was managed expectantly over the next several days with slow clinical and radiographic improvement. CONCLUSIONS: The etiology for the increased intraventricular pneumocephalus is believed to be retrograde leakage of air into the ventricles via the VP shunt during insufflation of the abdomen for percutaneous placement of a gastrojejunostomy feeding tube. PMID- 16378872 TI - Surgical treatment of pial cerebellar arteriovenous fistulas with aneurysm of the main feeding artery. AB - BACKGROUND: Intracranial pial arteriovenous fistulas (pAVFs) are rare vascular lesions only recently considered distinct from arteriovenous malformations. Conservative management was associated with a high mortality rate. The abnormality of the lesion arises from its high-flow nature. The authors present a case of a cerebellar pAVF associated with aneurysms of the main feeding artery that was surgically treated. CASE DESCRIPTION: A 54-year-old woman was referred to us after an attack of severe headache and vomiting. Computerized tomography scan visualized a hematoma of the fourth ventricle. Digital subtraction angiography (DSA) revealed a cerebellar pAVF mainly fed by the right posteroinferior cerebellar artery (PICA) and with drainage into one of the superior cerebellar veins. The PICA presented a saccular aneurysm on its fourth segment and a fusiform dilatation on its third segment. The patient underwent surgical operation via a midline suboccipital craniotomy. The fusiform dilatation was wrapped and the saccular aneurysm was clipped. The most fistulous connections were closed. The draining vein at the level of torcular herophili was closed by application of a clip. Control DSA demonstrated exclusion of both the aneurysms and the fistula from the circulation. At 1-year follow-up, the patient was neurologically intact. CONCLUSIONS: The association of a cerebellar pAVF and an aneurysm is rare. Surgical treatment of multiple-channel arteriovenous fistulas by closure of venous varix should guarantee the exclusion from the circulation and avoid recruitment of new arterial connections and recurrence. Attention must be paid to normal venous channels draining into the varix. PMID- 16378873 TI - Helmetlike skull deformity with a large arachnoid cyst. AB - BACKGROUND: It is not difficult to find localized skull ballooning or macrocrania in patients with intracranial arachnoid cysts. However, there have been no previous reports regarding large localized skull protuberant deformities resembling a war helmet. The authors report with a review of literature a case of an adult with helmetlike skull deformity resulting from a large supratentorial arachnoid cyst. CASE DESCRIPTION: A 35-year-old man presented with a large head deformity since his early childhood that had been the result of gradual progression from infantile macrocrania. He also had mental retardation, sixth cranial nerve palsy with recent aggravation of headache, reduced activity, poor voiding control, and walking disturbance. Magnetic resonance imaging of the head showed hydrocephalus with a large supratentorial arachnoid cyst located in the bilateral parietooccipital area compressing the hemisphere anteriorly, and the tentorium and cerebellum inferiorly. Magnetic resonance venogram demonstrated low lying short transverse and lateral sinuses, and the superior sagittal sinus and falx were displaced to the right side. Radioisotopic cisternogram showed nonfilling of the isotope in the bilateral parietooccipital area. Cerebrospinal fluid pressure measured by lumbar puncture was 17 cm H(2)O. We tentatively diagnosed the condition as normopressure hydrocephalus with a large supratentorial arachnoid cyst. His headache, reduced activity, poor voiding control, and walking disturbance improved after a cystoperitoneal shunt. CONCLUSIONS: This might suggest that large arachnoid cysts found in childhood should be treated for prevention of skull deformity and late aggravation of increased intracranial pressure. PMID- 16378875 TI - Odyssey between the constellations and neuromedicine. AB - The constellations have some connections with neuromedicine. As neurosurgeons, we can learn something from the night sky. Medical terms or descriptions are generously used from mythology or celestial lore. Neurosurgeons spend so much time on their professional careers that they may ignore something wonderful which is worthy of their attention. The puzzling issues between the constellations and neuromedicine can be exciting. Imagination allows neurosurgery to make great strides and progression. With that same imagination we can take an odyssey! PMID- 16378876 TI - How can you change direction in neurosurgery to adapt to the changes in our specialty? Strategies for your future. PMID- 16378877 TI - Whatever happened to the doctor's bag? PMID- 16378878 TI - Brugada-like electrocardiographic pattern: a challenge for the clinician. AB - A right bundle branch block with ST-segment elevation in the V1-V3 leads in characteristic coved or saddleback configuration may be encountered as an incidental finding. However, not all patients with a Brugada-like electrocardiographic pattern are affected by the Brugada syndrome; in fact, this pattern may also be found in healthy individuals. Whether symptomatic patients affected by the syndrome are at a high risk of developing life-threatening ventricular arrhythmias and should immediately receive an automatic, implantable defibrillator is open to debate, as is the clinical management of asymptomatic patients, because data from the scientific literature are controversial. Implications of the diagnosis and the treatment of this category of patients are discussed. PMID- 16378879 TI - Management of cirrhotic ascites: physiological basis of diuretic action. AB - Ascites is a significant complication of cirrhosis that occurs in approximately 50% of patients. The mortality rate is high in patients with cirrhosis and ascites. Conventional interventions rest with dietary sodium restriction, diuretic use, large-volume paracentesis, peritoneovenous shunts and transjugular intrahepatic portosystemic shunts. The mainstay of therapy, however, is the judicious use of diuretics. This article reviews the physiological basis of diuretic use in patients with cirrhosis and ascites, as well as recent concepts on the pathogenesis of ascites formation. Through a better understanding of the pathophysiology of ascites formation and the mechanism of action of diuretics, improved extracellular fluid balance can be achieved in these patients. PMID- 16378880 TI - Readmissions to hospital within 30 days of discharge from the internal medicine wards in southern Israel. AB - INTRODUCTION: Hospital readmission within a short time of discharge is a common phenomenon in internal medicine. Although there are a few reports of intensive care and surgical readmission rates there is almost no information available on the readmission rates of internal medicine patients. AIMS: To explore the medical patient readmission rate within 30 days of discharge from a medical center in southern Israel, and determine how it may be related to age, gender, diagnoses and other factors. METHODS: Chart review of all 2469 patients admitted to the departments of internal medicine of Barzilai Health Center between May 1st and July 15th 2002. A randomized control group of 87 patients was gathered from patients admitted to hospital during the study period but who were not readmitted within 30 days of discharge. RESULTS: 124 patients (5%) were readmitted. They were significantly older than controls (mean age 68.7+/-14.8 vs. 59.3+/-16.3 years), (p<0.0003), 21% being over 80 years of age (compared with 9% of controls, p<0.05) and had a lower average Functional Independence Measure (FIM). The index hospitalization of readmitted patients was significantly longer than the hospitalization of control patients (6.1+/-5.5 vs. 4.1+/-3.8 days), (p<0.04). Most patients (85%) were readmitted for the same medical reason that prompted the original hospitalization. The 15% readmitted for other reasons were older (74.9+/ 12.4 vs. 67.6+/-15.0 years) (p<0.001) and had a lower Functional Independence Measure (mean FIM 4.6+/-2.2 vs. 5.7+/-1.9), (p<0.05). New immigrants (mean age 71.1+/- 12.9 years) had significantly lower readmission rate compared to the general population (3.2% vs. 4.98%), (p<0.05). Veteran Israelis, on the other hand, had a higher readmission rate than the general population (6.8% vs. 4.98%) (p<0.001). CONCLUSION: Only 5% of patients required readmission. The original hospitalization period of readmitted patients was significantly longer than controls and most readmissions were for the original medical problem for which they were readmitted originally or because of a low functional status. Hospitalized new immigrants although being older in age are less likely to require readmission, while veteran Israelis are more likely to be readmitted. PMID- 16378881 TI - Snake venom poisoning in Greece. Experiences with 147 cases. AB - BACKGROUND: Snake venom poisoning is a medical emergency requiring immediate attention. Bites from poisonous European snakes can lead to local tissue damage and systemic symptoms. Vipera ammodytes accounts for the most envenomation in Greece. METHODS: The demographic and epidemiological characteristics, clinical symptoms and signs, laboratory findings, treatment, and outcome of 147 consecutive victims of V. ammodytes admitted to our hospital from 1988 to 2003 were reviewed and analyzed. RESULTS: The most common symptoms and signs included fang marks (100%), pain (100%), swelling (98.64%), ecchymosis (60.54%), tachycardia (32.65%), fainting or dizziness (29.93%), fever (23.13%), enlargement of regional lymph nodes (17.69%), nausea (16.33%), hypotension (13.61%), and vomiting (12.93%). The main complications were reduced range of motion, thrombophlebitis, local hemorrhagic blister formation, skin bleeding, rhabdomyolysis, reduced sensation, acute renal failure, necrosis with tissue loss, carpal tunnel syndrome, compartment syndrome, Kounis syndrome, and digit amputation. CONCLUSIONS: A V. ammodytes bite is a potentially serious event that requires immediate hospital care. Yet, the majority of victims can be treated successfully with conservative methods. No deaths occurred in our series. PMID- 16378882 TI - Does this patient have cancer? The assessment of age, anemia, and erythrocyte sedimentation rate in cancer as a cause of weight loss. A retrospective study based on a secondary care university hospital in Romania. AB - BACKGROUND: "Does this patient have cancer?" is a question frequently asked when confronted by patients with involuntary weight loss. The aim of this study was to assess the value of age, erythrocyte sedimentation rate (ESR), and anemia in the diagnosis of cancer as a cause of involuntary weight loss. METHODS: A retrospective study of 7850 patients admitted to the Department of Internal Medicine from January to September 2003 was performed. Especially selected were 431 patients with weight loss. Age, ESR, hemoglobin, and the discharge diagnosis were recorded. RESULTS: Twenty-four percent of the patients with involuntary weight loss had cancer. Age, ESR, and anemia were found not to be of value in the diagnosis of cancer (areas under the curve were 0.684, 0.690, and 0.766, respectively). When diagnostic tests for age, a high ESR, and anemia were used serially, the positive predictive value for a malignancy was 64% (CI: 27-90%); when the tests were utilized in parallel, the negative predictive value was 91% (CI: 85-100%). CONCLUSIONS: Any patient admitted to our Department of Internal Medicine for involuntary weight loss had a 24% probability of having a malignancy. Neither age, nor ESR, nor anemia, used separately as a multilevel, diagnostic test or combined serially or in parallel, could exclude or rule in the diagnosis of cancer. However, they could increase (from 24% to 64%) or decrease (from 24% to 9%) the probability of cancer. PMID- 16378883 TI - Attitudes to, and practice of, unconventional medicine by physicians in Italy. AB - BACKGROUND: Given the great public demand for unconventional medicines (UM) in most Western countries, the aim of this study was to assess the attitudes to, and supply of, UM by physicians in Italy. METHODS: A cross-sectional survey of all of the physicians belonging to the Ordine dei Medici of the province of Parma was carried out by means of an anonymous questionnaire mailed to 2631 physicians and returned by 1734 (66%). The outcome measurements were the prevalence of opinions concerning UM, the prevalence of its practice, and the extent to which demographic and practice characteristics influenced it. RESULTS: The majority of the physicians (53%) attributed some efficacy to UM. This belief was significantly more frequent amongst female physicians (p<0.01). A small proportion of physicians (8%) claimed to practice some form of UM. The following types of physicians were also more likely to practice UM: rural physicians (p=0.01), those working as National Health Service (NHS) general practitioners or in private practice (p<0.0001) and those specialized in areas relating to the musculoskeletal apparatus or anesthesia/intensive care (p<0.01). CONCLUSIONS: This Italian survey found a smaller proportion of physicians practicing UM on their patients than those indicated by other published surveys. PMID- 16378884 TI - Plasma markers of endothelial dysfunction in type 2 diabetics. AB - BACKGROUND: Type 2 diabetes, or non-insulin-dependent diabetes mellitus, represents an independent risk factor for cardiovascular diseases (CVD), being characterized by a continuous low-grade inflammation and endothelial activation state. Atherosclerotic lesions occur in diabetic patients at an earlier age with severe clinical manifestations and poor outcome. Our objective was to investigate the correlation between lipoprotein-associated phospholipase A2 (PLA2-LDL), myeloperoxidase (MPO), and paraoxonase (PON), enzymes implicated in the evolution of endothelial dysfunction associated with type 2 diabetes. METHODS: One hundred diabetic patients [50 without documented coronary artery disease (group 1) and 50 with CVD (group 2)] and 46 healthy controls were investigated for PLA2-LDL, MPO, and PON activities. RESULTS: PLA2-LDL activity was significantly higher in group 2 than in group 1 and among controls. PON activity was lower in group 1 than in controls, reaching the lowest level in group 2. MPO activity was higher in type 2 diabetics than among controls, with similar values in groups 1 and 2. CONCLUSIONS: The evaluation of PLA2-LDL, MPO, and PON activities may improve early diagnosis of CVD in asymptomatic patients with type 2 diabetes and can help to evaluate accelerated atherosclerosis and microvascular disease. PMID- 16378885 TI - Disagreement in the interpretation of chest radiographs among specialists and clinical outcomes of patients hospitalized with suspected pneumonia. AB - BACKGROUND: Pneumonia is diagnosed by a combination of clinical symptoms and findings on chest X-ray (CXR). However, there is often disagreement, even among experts, upon the interpretation of the CXR. The purpose of this study was to compare the agreement rates in CXR interpretation of suspected community-acquired pneumonia (CAP) between a radiologist, a pulmonologist, an infectious disease specialist, and an internal medicine staff and to establish the correlation of such an agreement with the length of hospitalization and 30-day mortality rate. METHODS: We prospectively enrolled in our study all patients admitted to the Department of Medicine with suspected CAP, as defined by the admitting physician, within the first 24 h of hospitalization. A radiologist, pulmonologist, and infectious disease specialist who were aware of the suspected diagnosis independently interpreted the CXR. The final diagnosis was obtained from the discharge notes. RESULTS: A total of 262 patients participated in the study, 214 of whom (81.7%) were eventually discharged with a diagnosis of CAP. The agreement rates between the readers of the CXR ranged from a kappa of 0.09 to 0.44. There were no differences in terms of background illness, PORT (Pneumonia Patients Outcomes Research Team) score, length of hospitalization, or mortality rates between patients discharged with or without a diagnosis of CAP. In multivariate analysis, only the PORT score was a significant predictor of length of hospitalization and mortality rate. CONCLUSION: We found a low to moderate agreement rate of the diagnosis of CAP between CXR readers. Identification of an infiltrate on CXR, either by specialists or by the attending physician, did not impact the clinical outcomes. PMID- 16378886 TI - Primary prevention of arterial thromboembolism in the oldest old with atrial fibrillation--a randomized pilot trial comparing adjusted-dose and fixed low-dose coumadin with aspirin. AB - BACKGROUND: Anticoagulation therapy remains largely underused in older people with atrial fibrillation (AF). The aim of this pilot trial was to investigate the effects of low-intensity adjusted-dose and fixed-dose coumadin on the incidence of arterial thromboembolism and bleeding, as well as the efficacy of anticoagulation monitoring in elderly AF patients. METHODS: In this open-label, randomized trial we recruited patients over 75 years of age without previous stroke or systemic embolism. Patients were randomized into three groups, with group A receiving aspirin 100 mg/day, group B fixed-dose coumadin 1 mg/day; and group C adjusted-dose coumadin with a target range of international normalized ratios (INR) between 1.6 and 2.5. Primary endpoints (ischemic strokes and systemic embolisms) and secondary endpoints (deaths, myocardial infarctions, and major bleeding events) were prospectively documented. RESULTS: The study was discontinued 6 months after the enrollment of the first patient for safety reasons. During this period, 45 patients were recruited (15 patients in group A, 14 in group B, and 16 in group C). Over a mean follow-up period of 3.7 months (range: 1-6 months), two patients from group B developed a dangerous prolongation of the INR (7.0 and 4.2), which led to the discontinuation of fixed-dose coumadin. Another patient from the same group experienced a major bleeding event 1 month after enrollment in the study (INR: 5.5). The percentage of INR measurements within the target range was significantly (p<0.001) lower in group B (48.7%) than in group C (83.7%). CONCLUSIONS: Older people receiving fixed-dose oral anticoagulants may be at risk of developing a dangerous prolongation of their INR. PMID- 16378887 TI - Tracheal stenosis diagnosed on pulmonary ventilation/perfusion scan. AB - A 68-year-old woman, who had undergone coronary artery bypass surgery 5 months previously, was presented with cough, breathlessness and an elevated D-dimer. She was initially thought to have suffered a pulmonary embolus. A ventilation/perfusion scan demonstrated tracheal stenosis, which required dilation and endobronchial stent deployment. Tracheal stenosis is a well recognised complication of endotracheal intubation; however, the onset of symptoms is often insidious and the diagnosis delayed. PMID- 16378888 TI - To biopsy or not to biopsy: an 82-year-old patient with a retroperitoneal mass and severe low back pain. AB - An 82-year-old patient was admitted to our ward due to disabling severe low back pain. Computed tomography of the spine revealed a retroperitoneal space-occupying lesion encroaching on two adjacent lumbar vertebrae and causing destruction of the cortex of their anterior aspect. The patient was scheduled for a biopsy of the mass. Magnetic resonance (MR) of the lumbar spine, however, suggested that the mass was most probably an aortic aneurysm. The biopsy was cancelled and the patient was referred for surgical intervention. MR is indicated in the evaluation of a solid mass causing vertebral destruction in order to achieve an accurate preoperative diagnosis and prevent a hazardous invasive procedure. PMID- 16378889 TI - Primary Sjogren's syndrome associated with pneumatosis cystoides coli. AB - We report the case of a 43-year-old woman with primary Sjogren's syndrome, according to Americano-European criteria. Eighteen months after the diagnosis, the patient presented pneumatosis cystoides coli (PCC), which resolved with medical treatment consisting of diet and cisapride. Four years after this episode, the patient has not developed clinical features of another systemic inflammatory rheumatic disease and PCC has not relapsed. To the best of our knowledge, the association between primary Sjogren' syndrome and PCC has never been reported. Physiopathology and treatment of PCC are discussed. PMID- 16378890 TI - A case of intravascular lymphomatosis (IVL) presenting as polyneuropathy. AB - Intravascular lymphomatosis (IVL) is a rare, yet important, cause of a variety of unexplained neurological syndromes. In this clinical context, the only clue to the diagnosis may be a significantly raised plasma lactate dehydrogenase (LDH) on a background of clinical features of a systemic illness. PMID- 16378891 TI - CT evaluation of renal artery embolism with ectopic artery. AB - We present an 80-year-old woman, with a history of well-controlled hypertension and atrial fibrillation, who suffered from back pain and elevated blood pressure for 2 weeks prior to admission. At physical examination, a low-pitched systolic heart murmur was heard in the tricuspid area and there was mild tenderness in the left lower quadrant. Laboratory studies revealed an elevated hematocrit, lactate dehydrogenase, and liver aminotransferase. Macroscopic hematuria was present in urine and 20% eosinophils were detected with Hansel's stain method. Assuming a possible vascular embolic episode, angiography of the renal arteries was performed, which revealed an obstructive pattern in the presence of a left duplicate system. PMID- 16378892 TI - Pulmonary mucormycosis: two of a kind. AB - Mucormycosis is an uncommon infection caused by fungi of the class Zygomycetes, suborder Mucorales. It occurs predominantly in the immunodeficient host, the most common risk factor being diabetes mellitus. Pulmonary mucormycosis is a disease that is localized in the lungs or mediastinum. It usually presents in a fulminant manner, although a chronic presentation can also occur. We describe two cases of pulmonary mucormycosis in patients with diabetes mellitus, one with an acute and the other with a chronic presentation. A high level of clinical suspicion, combined surgical and medical therapy, and control of patient's underlying immunosuppression are the key to a successful outcome. PMID- 16378893 TI - Fulminant Budd-Chiari syndrome associated with polycythemia rubra vera and factor V Leiden mutation. AB - Budd-Chiari syndrome (BCS) is a severe disorder characterized by hepatic venous outflow obstruction. Hypercoagulable states are the major etiological factors for the development of BCS and can be identified in about 75% of patients. Multiple etiological factors can be found in the same patient. Hematologic abnormalities, especially myeloproliferative disorders, are the most common causes of BCS. Furthermore, the prevalence of factor V Leiden mutation is three times greater in patients with BCS. Although the clinical course tends to be chronic, BCS may, on rare occasions, cause acute liver failure. Herein, we report a patient who had factor V Leiden mutation and polycythemia rubra vera, presented as fulminant BCS. PMID- 16378895 TI - Valproic acid-associated eosinophilic pleural effusion treated with corticosteroids. PMID- 16378894 TI - A rare cause of fatal right heart failure. AB - Microscopic pulmonary tumor embolism (MPTE) is an uncommon cause of dyspnea in patients with cancer and one of the most difficult to diagnose. MPTE is a syndrome that is pathologically characterized by the occlusion of small pulmonary arteries and arterioles by aggregates of tumor cells. Because the clinical picture resembles that of thromboembolic disease, it is rarely recognized before death. The most common clinical symptom is subacute progressive dyspnea over weeks to months. We recently observed a case of MPTE of exceptional interest as the patient was under aggressive anticoagulant treatment and developed fulminant pulmonary hypertension with fatal right heart failure. PMID- 16378896 TI - Late left ventricular perforation as a complication of permanent pacing leads. PMID- 16378897 TI - Angiotensin converting enzyme inhibitor toxicity causing interstitial pneumonitis and cholestatic hepatitis. PMID- 16378898 TI - Sarcoidosis masquerading as ankylosing spondylitis. PMID- 16378900 TI - Introduction. PMID- 16378901 TI - Biologic basis for combining drugs with radiation. AB - Concurrent administration of radiation and chemotherapy has emerged as a dominant form of cancer treatment. Nevertheless, our understanding of the specific mechanisms of interaction between radiation and chemotherapy is still evolving. Biological evidence gained in experimental cell lines and tumors suggests that cisplatin lesions cause repair inhibition of radiation-induced DNA damage affecting both homologous recombination and nonhomologous end joining. In the case of 5-fluorouracil (5-FU), the crucial event in the radiosensitisation process appears to be inappropriate progression through S-phase in the presence of bound drug affecting both DNA repair and cell cycle. In similar fashion, data with gemcitabine suggest that radiosensitization arises through progression into S-phase causing depletion of 2'-deoxyadenosine 5'-triphosphate pools, which leads to misincorporation and misrepair of incorrect bases in irradiated cells. As the new molecular-targeted agents become increasingly available to complement established radiation and chemoradiation treatment regimens, new concepts of drug radiation interaction are evolving that require innovative thinking regarding the manner in which radiation and drugs cooperate and how clinical trials are best performed. PMID- 16378902 TI - Platinum-based concurrent chemoradiotherapy for tumors of the head and neck and the esophagus. AB - The addition of concurrent chemotherapy (CT) to standard radiotherapy (RT) for locoregional treatment has been established to improve overall survival in a variety of solid tumors. Among the many CT regimens evaluated in combination with RT in randomized controlled clinical trials and summarized in meta-analyses, platinum-containing regimens have consistently shown a survival benefit across tumor types. Cisplatin and carboplatin have been studied both as single agents and in combination with other cytotoxic drugs, concurrently with RT, but the optimal platinum-based regimen to be combined with RT continues to be explored with further investigation. In this article, the role of platinum-based CT as part of concurrent CT/RT will be discussed using 2 tumor sites in the aerodigestive tract as a paradigm: squamous-cell carcinomas of the head and neck and esophageal carcinomas. For each tumor type, we will review the state of the evidence and comment on the current state of practice and on future directions for clinical research in combined modality CT/RT. PMID- 16378903 TI - Radiation therapy combined with hormone therapy for prostate cancer. AB - Standard-dose radiation therapy has limited capacity to cure bulky and locally advanced prostate cancer. Multiple randomized trials have shown a clinical benefit to adding androgen suppression therapy to external-beam radiation therapy in several subsets of prostate cancer. These studies have made combining hormonal therapy with radiation therapy the standard of care for men with locally advanced (T3-4) and unfavorable prostate cancers (Gleason score >or=8 and/or prostate specific antigen >20 ng/mL). The clinical impact of hormonal therapy has been seen in biochemical control, local control, distant metastases, disease-specific survival, and overall survival. If hormonal therapy is to be combined with radiation, it should be initiated before the start of radiation and continued during the radiation course rather than used only in the adjuvant setting. Typically, shorter-term hormone therapy is defined as regimens of 4 to 6 months, with longer-term hormone therapy describing durations beyond 24 months. Historically, longer-term hormone therapy was thought to have a more profound systemic effect; however, with the emerging use of hormonal therapy for less advanced disease, the overall impact of shorter-course hormone therapy is being seen. This review will summarize trials using hormonal therapy and radiation with an emphasis on phase III studies and describe the more recent integration of hormone therapy with radiation for prostate cancer. PMID- 16378904 TI - Novel radiation-enhancing agents in malignant gliomas. AB - Malignant gliomas represent among the most aggressive of all known human neoplasms. These tumors show high levels of resistance to conventional therapies (ie, surgery, radiation, and chemotherapy). For many decades, significant improvements in clinical outcomes of newly diagnosed glioblastoma (GBM) patients have not been observed until recently. A landmark European Organization for Research and Treatment of Cancer (EORTC) phase III study showed significant improvements in median and 2-year survivals in newly diagnosed GBM patients treated with a combination of radiation and temozolomide chemotherapy versus radiation alone. The results of this study have certainly defined practice standards for this patient population. This article will review the role of both traditional nitrosurea-based and newer-generation chemotherapeutic and biotherapeutic agents when combined with radiation therapy for the treatment of malignant glioma patients. PMID- 16378905 TI - Radiation combined with EGFR signal inhibitors: head and neck cancer focus. AB - Molecular inhibition of epidermal growth factor receptor (EGFR) signaling represents one of the most promising current arenas for the advancement of molecularly targeted cancer therapies. A series of EGFR inhibitors from both the monoclonal antibody (mAb) and tyrosine kinase inhibitor (TKI) class have shown clear clinical activity in the treatment of several common human cancers. Three EGFR inhibitors have recently gained Food and Drug Administration (FDA) approval for cancer therapy in the United States including the mAb cetuximab (Erbitux) and the small-molecule TKIs gefitinib (Iressa) and erlotinib (Tarceva). The rapidly expanding preclinical and clinical data contributing to these FDA approvals validate a central role of the EGFR as an important molecular target in epithelial malignancies. Indeed, one of the more striking clinical results in this field has been recently achieved by combining an EGFR inhibitor (cetuximab) with radiation in the treatment of advanced head and neck cancer patients. Nevertheless, the overall clinical gains realized to date with the EGFR inhibitors are modest for the global cancer population. Much remains to be learned regarding the rational integration of EGFR inhibitors into cancer treatment regimens as well as methods to optimize the selection of patients most likely to benefit from EGFR inhibitor strategies. PMID- 16378906 TI - Radiation combined with antiangiogenic and antivascular agents. AB - Angiogenesis is critical for a number of physiologic and pathophysiologic processes, and angiogenesis inhibitors are now being used in the treatment of cancer. Although antiangiogenic agents offer great therapeutic potential, preclinical and clinical trial results suggest that these agents will have a delayed onset of activity and may only induce disease stabilization for patients with advanced malignancy. The use of radiation therapy for cancer is also associated with therapeutic challenges that are distinct from those that might be expected with antiangiogenic agents. Thus, the use of angiogenesis inhibitors in combination with radiation therapy should help to overcome the limitations of each leading to enhanced efficacy and diminished toxicity. The goal of this review is to provide an overview and discussion of the combination of angiogenesis inhibitors with radiation therapy. PMID- 16378907 TI - Radiation and new molecular agents part I: targeting ATM-ATR checkpoints, DNA repair, and the proteasome. AB - In response to DNA breaks, human cells delay their progression through the G1, S, and G2 phases of the cell cycle. This response requires the coordinated effort of the ATM-CHK2-p53 and ATR-CHK1 DNA damage-sensing pathways and DNA repair (eg, DNA PK and RAD51 complexes). The turnover of many of these DNA damage-associated proteins is controlled by the 26S proteasome. In this article, we review molecular strategies that target each of these pathways using silencing RNA (siRNA), antisense, or small-molecule inhibition. Although these agents can radiosensitize tumor cells, little data are available regarding potential effects on normal tissues to determine the potential therapeutic ratio of these strategies after fractionated radiotherapy. Clinical trials using such agents will require novel correlative science endpoints to track DNA repair and cell cycle arrest and will need careful assessment of normal tissue toxicity and stability. PMID- 16378908 TI - Radiation and new molecular agents, part II: targeting HDAC, HSP90, IGF-1R, PI3K, and Ras. AB - Current research efforts in cancer therapeutics include the development of novel inhibitory agents that target molecular pathways involved in tumor growth and progression. Ultimately, many of these agents may prove most efficacious when combined with conventional cytotoxic therapies, including radiation therapy. Elucidation of the biologic pathways underlying radiation response has identified several targets involved in radiation resistance, providing rationale for combining these agents with radiation. Agents targeting single pathways, including EGFR, IGF-1R, PI3K, and Ras, have been studied alone and in combination with radiation. Although this strategy is increasingly supported by preclinical and clinical data, the single-target approach may be limited by such factors as tumor heterogeneity and genetic instability. Emerging approaches include multipathway-targeted therapy by either combining target-specific agents or using single agents that target multiple pathways, including HDAC and HSP90 inhibitors. These approaches reviewed herein hold promise for improved radiation therapy efficacy and, ultimately, improved patient outcome. PMID- 16378909 TI - Complementary and alternative therapies: the power of randomization. PMID- 16378910 TI - Elevated second trimester amniotic fluid interferon gamma-inducible T-cell alpha chemoattractant concentrations as a possible predictor of preterm birth. AB - OBJECTIVE: To determine and correlate midtrimester amniotic fluid concentrations of interferon gamma-inducible T-cell alpha chemoattractant (ITAC, a chemokine directing the migration of activated T lymphocytes toward inflammation sites) and C-reactive protein (CRP) in women undergoing amniocentesis and subsequently delivering pre-or full-term infants. METHODS: Among 312 women undergoing midtrimester transabdominal amniocentesis, 13 progressed to spontaneous delivery at less than 37 gestational weeks (GW). Subjects were matched for maternal age, parity, and GW at amniocentesis with 21 controls who delivered at greater than 37 GW. Amniotic fluid ITAC and CRP levels were determined by enzyme-linked immunosorbent assay (ELISA) and by nephelometry, respectively. RESULTS: Both ITAC and CRP values were significantly higher (P = .005 and P = .04, respectively) in the amniotic fluid of women delivering at less than 37 GW. A statistically significant correlation between amniotic fluid ITAC and CRP concentrations was also found (r = .366, P < .05). The receiver operator curve (ROC) analysis of delivery at less than 37 GW gave the best cutoff point for ITAC at a concentration of 44 pg/mL and for CRP at a concentration of 0.16 mg/dL. Positive and negative predictive values for ITAC were 82% and 85%, respectively, and for CRP, 55% and 76%, respectively. CONCLUSIONS: Present data indicate that from the second trimester of pregnancy elevated amniotic fluid concentrations of ITAC are found in women delivering at less than 37 GW, as compared to women delivering at term. Therefore, ITAC in combination with other cytokines or CRP could possibly serve as predictor of preterm delivery. PMID- 16378911 TI - TASK channel expression in human placenta and cytotrophoblast cells. AB - OBJECTIVE: The multinucleate syncytiotrophoblast is the transporting epithelium of the human placental villus, formed throughout pregnancy by fusion and differentiation of underlying mononucleate cytotrophoblast cells. Similar to other epithelia, K+ channels will impact on syncytiotrophoblast transport properties during its development and differentiation. Therefore we investigated expression and activity of the two-pore domain K+ channels TASK1 and 2 in relation to gestation and differentiation, using villous tissue from first and third trimester and cultured cytotrophoblast cells at mononucleate and multinucleate stages of culture. METHODS: Quantitative real-time polymerase chain reaction (PCR), immunofluorescence, and 86Rb+ (K) efflux were used to investigate TASK channel expression and function. RESULTS: TASK2 mRNA expression was higher in first trimester than term (10 to 13 vs 38 to 40 weeks, P < .05). Other K+ alpha-subunit mRNAs, including TASK1, remained unaltered but the regulatory BKCa beta-subunit, like TASK2, was higher in first trimester than term (P < .001). Immunofluorescence showed that TASK2 had an intracellular localization within the trophoblast of first trimester villi but was less abundant and restricted to stem villi at term. TASK2 also showed intracellular localization in mononucleate cytotrophoblast cells in culture and expression was lost with multinucleation. By contrast, TASK1 was localised, independently of cell nucleation, to cytotrophoblast cell plasma membranes. 86Rb+ (K) efflux was measured from multinucleated cytotrophoblast cells. Both basal and pH 8.0-stimulated efflux was inhibited by the TASK1 antagonist anandamide (n = 5 for both conditions; P < .01 and P < .001, respectively). CONCLUSION: TASK1 and 2 are expressed in placental trophoblast cells and TASK1 activity may have a role in regulating syncytiotrophoblast homeostasis and/or solute transport functions. PMID- 16378912 TI - Low plasma volume coincides with sympathetic hyperactivity and reduced baroreflex sensitivity in formerly preeclamptic patients. AB - BACKGROUND: Preeclampsia is associated with enhanced sympathetic activity as well as subnormal plasma volume. Meanwhile, in over 50% of these complicated pregnancies, the subnormal plasma volume has been found to persist for a prolonged period after pregnancy. The objective of this study is to test the hypothesis that in normotensive formerly-preeclamptic women, persistence of a subnormal plasma volume coincides with enhanced sympathetic activity and with it, an altered autonomic control of blood pressure. METHODS: Forty-eight formerly preeclamptic women participated in this study. After measurement of their plasma volume by iodine 125-albumin indicator dilution, they were subdivided into a group with a normal plasma volume (plasma volume > 48 ml/kg lean body mass) and a group with a subnormal plasma volume (< or = 48 ml/kg lean body mass). We performed spectral analysis on their beat-to-beat blood pressure and heart rate recordings and compared both groups using non-parametric tests. RESULTS: Formerly preeclamptic women with a subnormal plasma volume had a higher sympathetic activity (P = .001) and a lower baroreflex sensitivity (P = .04) than their counterparts with a normal plasma volume. CONCLUSION: In normotensive formerly preeclamptic women, a subnormal plasma volume coincides with a higher sympathetic activity in the blood pressure regulation and lower baroreflex sensitivity. Whether these alterations in the autonomic control mechanisms are a cause or effect of the subnormal plasma volume remains to be elucidated. PMID- 16378913 TI - Maternal interleukin-6: marker of fetal growth and adiposity. AB - Fetal overgrowth and higher adiposity are hallmarks of pregnancy with maternal obesity and poor glucose tolerance, two conditions associated with decreased maternal insulin sensitivity. In non-pregnant individuals, adipokines, vasoactive peptides, and components of the immune system crosstalk with metabolic factors to generate signals triggering obesity and impaired insulin action. We have investigated circulating maternal and fetal cytokines and growth-factors as potential biochemical markers of fetal adiposity. Mothers and neonates were classified into three tertiles (T1-T3) using total neonatal fat mass as the outcome with 309 +/- 25 g in T1, 478 +/- 40 g in T2, and 529 +/- 39 g in T3. Umbilical cord endothelin-1 (ET-1), C-peptide, and leptin were higher in T3 and T2 versus T1. Only cord leptin was strongly associated with fetal fat mass (P < .01), whereas neonatal lean body mass was negatively correlated with maternal insulin-like growth factor binding protein-I (IGFBP-I) (r = -0.53, P < .04). This study shows an association between increased fetal adiposity and maternal systemic interleukin-6 (IL-6). No such correlation was found with factors circulating in cord blood, suggesting that the stimuli favoring fetal fat accretion derive from maternal or placental sources rather than from the fetus. PMID- 16378914 TI - Effect of peroxisome proliferator-activated receptor-gamma agonist rosiglitazone on the induction of endometriosis in an experimental rat model. AB - OBJECTIVE: To assess the effect of the peroxisome proliferator-activated receptor (PPAR)-gamma agonist rosiglitazone on the induction of endometriosis in a rat model. METHODS: Endometriosis was surgically induced in 28 rats by transplanting an autologous fragment of endometrial tissue onto the inner surface of the abdominal wall. Group I was assigned as control and no medication was administered. Starting 3 days before the operation and continuing for 4 weeks, 0.2 mg/kg/d rosiglitazone was administered to the study group orally. Four weeks later rats were killed and ectopic uterine tissues were evaluated morphologically and histologically. Scoring systems were used to evaluate preservation of epithelia. RESULTS: Four rats in the study group and one rat in the control group died of complications related to surgery. There was a significant difference in post-treatment spherical volumes (64.00 mm3 [interquartile range (IQR): 354.42] vs 41.60 mm3 [IQR: 37.87], P = .018) and explant weights (77.97 mg [IQR: 431.27] vs 47.24 mg [IQR: 43.01], P = .005) between control and rosiglitazone-treated groups. The epithelia were found to be preserved significantly better in the control group when compared with the roziglitazone-treated group (2.00 [IQR:2.00] vs 0.00 [IQR:2.25], P = .014). CONCLUSIONS: Rosiglitazone was found to affect the induction of endometriosis negatively in this experimental rat model and seemed to interfere with the growth and maintenance of the uterine explant. PMID- 16378915 TI - Effects of Chinese green tea on weight, and hormonal and biochemical profiles in obese patients with polycystic ovary syndrome--a randomized placebo-controlled trial. AB - OBJECTIVES: To study the effects of green tea on body weight, and biochemical and hormonal profiles in obese Chinese women with polycystic ovary syndrome (PCOS). METHODS: Thirty-four obese Chinese women with PCOS were randomized into either treatment with green tea capsules or placebo for 3 months. The anthropometric measurements, and biochemical and hormonal profiles before and after treatment in each group were compared. RESULTS: The body weight of the green tea group decreased by a nonsignificant 2.4% after treatment; whereas the body weight, body mass index (BMI), and body fat content of the control group were significantly higher after 3 months. There were no differences in any of the hormone levels measured in either group. The biochemical profiles of the two groups were also similar except that there was a small but significant rise in the triglyceride level in the green tea group. Fewer patients in the green tea group remained amenorrhoeic, but this was not significantly different from the control group. CONCLUSIONS: Green tea supplementation did not significantly reduce body weight in obese women with PCOS, nor did it alter the glucose or lipid metabolism. PMID- 16378916 TI - Expression of the transcriptional coregulator FHL2 in human breast cancer: a clinicopathologic study. AB - OBJECTIVE: Although the Four and a Half LIM domain protein 2 (FHL2) has been suggested to play an important role in tumor development, this has not been investigated in breast cancer. METHODS: Paraffin-embedded tissues from patients (n = 85) with primary breast cancer were submitted to immunohistochemical investigation of FHL2 expression and subsequent correlation with clinicopathologic parameters and patient survival. RESULTS: The expression of FHL2 was confined to the cytoplasm of the tumor cells. Forty (47%) of 85 samples showed weak expression of FHL2, whereas high expression was found in 45 tumors (53%). A statistically significant positive correlation was observed between FHL2 and androgen receptor expression (P = .029). Patients with tumors expressing low amounts of FHL2 were characterized by a significantly better survival compared to those with high intratumoral FHL2 expression (P = .0215, log-rank test). The additional stratification according to adjuvant tamoxifen treatment revealed a significantly improved survival rate for patients receiving tamoxifen and being diagnosed with a tumor expressing high amounts of FHL2. This might indicate that tamoxifen is at least partially capable of reversing the negative prognostic impact of high FHL2 expression. Multivariate Cox regression analysis revealed FHL2 expression as a significant independent predictor of survival. CONCLUSION: The specific expression in tumor tissue points to an important functional role of FHL2 in human breast cancer. Our survival data indicate that the expression of FHL2 in primary breast cancer is a potentially relevant prognostic factor. Further studies are warranted to elucidate whether analysis of FHL2 expression is suitable to predict response to antihormonal treatment with tamoxifen. PMID- 16378917 TI - Triphasic mitral inflow velocity with mid-diastolic flow: the presence of mid diastolic mitral annular velocity indicates advanced diastolic dysfunction. AB - Mitral inflow filling pattern usually consists of 2 forward flow velocities in sinus rhythm: early rapid filling (E) and late filling with atrial contraction (A). However, additional mid-diastolic flow velocity may be present resulting in triphasic mitral inflow filling pattern. When mitral inflow is triphasic, mitral annulus velocity recorded by tissue Doppler imaging (TDI) frequently demonstrates a mid-diastolic component (L'). The significance of L' has not been explored previously. The purpose of this study was to explore possible mechanisms and clinical implications of triphasic mitral inflow with or without L' using TDI and proBNP. Of 9004 patients who underwent transthoracic echocardiography from March to November 2003, 83 (0.9%) patients (33 male, 50 female; mean age, 63+/-10 years) with a triphasic mitral inflow velocity pattern, including mid-diastolic flow velocity of at least 0.2m/s, and sinus rhythm were prospectively identified in our clinical echocardiography laboratory. Peak velocity of E, mid-diastolic (L), and A, and deceleration time (DT) of the E wave velocity were measured. Diastolic mitral annular velocities were measured at the septal corner of the mitral annulus by TDI from the apical 4-chamber view. ProBNP was measured at the time of echocardiogram using a quantitative electrochemiluminescence immunoassay. Mean heart rate was 54+/-6 beats/min (range, 40-67). Mean left ventricular (LV) ejection fraction (EF) was 64+/-13% and LV systolic dysfunction (EF<40%) was present in only 6 (7%). Patients were classified into 2 groups: group 1 (n=47) included those who had L' and group 2 (n=36) included those without L'. Group 1 patients had significantly higher peak velocity (35+/-14 vs 26+/-6 cm/s, p=0.0002) and TVI (35+/-14 vs 26+/-6 cm/s, p=0.0002) of L, E/E' (18+/-8 vs 14+/ 6, p=0.02), and left atrial volume index (42+/-14 vs 34+/-10 ml/m(2), p=0.0037). E' (4.7+/-1.3 vs 6.2+/-2.3 cm/s, p=0.001) and A' (6.2+/-2.0 vs 8.6+/-3.4 cm/s, p=0.0006) were significantly lower in group 1 compared with those of group 2. ProBNP was significantly higher in group 1 (847+/-1461 vs 438+/-1039 pmol/l, p=0.0012) and it was above normal in all except in 1 patient of group 1. In conclusion, the presence of L' in subjects with triphasic mitral inflow velocity pattern with mid-diastolic flow is associated with higher E/E', elevated proBNP and enlarged left atrium indicating advanced diastolic dysfunction with elevated filling pressures. This unique mitral annular velocity pattern should be helpful in identifying the patients with advanced diastolic dysfunction and increased LV filling pressures. PMID- 16378918 TI - Effects of cardiac resynchronization therapy on the mechanisms underlying functional mitral regurgitation in congestive heart failure. AB - AIMS: Functional mitral regurgitation (FMR) improvement induced by cardiac resynchronization therapy (CRT) has been related to left ventricular (LV) remodeling reversal and contractility enhancement. The effects induced by the changes of LV synchronicity indexes on FMR severity have not been investigated. METHODS AND RESULTS: In 30 patients with CRT for heart failure (HF) and QRS>130 ms, LV function parameters, FMR severity as mitral jet regurgitation/left atrial area ratio (JA/LAA) and standard deviation (SD) of the time to the systolic peak velocity at 6-basal and mid-LV segments as asynchrony indexes were evaluated (echo/tissue Doppler) before and 6 months after implant. At follow-up, 15 patients resulted responders to LV reverse remodeling with > or =15% end-systolic volume (ESV) and LV systolic function improvement. Improvement of FMR with > or =15% JA/LAA reduction was observed in 19 patients, 7 were nonresponders to LV reverse remodeling. In patients with > or =15% JA/LAA reduction a significant decrease of LV asynchrony indexes was observed as compared to patients without > or =15% JA/LAA reduction in whom LV asynchrony indexes were increased. Reduction of LV mid-segmental asynchrony was the variable most strongly related to JA/LAA reduction (r(2)=0.697, P<0.01), with good agreement between observed and predicted values (only 1 patient outside the mean+/-2SD). CONCLUSION: These data reveal that CRT can reduce FMR irrespective to LV remodeling reversal; this effect is related to LV asynchrony reduction and further support CRT employment in patients with HF and FMR. PMID- 16378919 TI - Intracardiac pseudotumor caused by mitral annular calcification. AB - The current report describes a rare case of a pseudotumor in the left ventricle. Transthoracic and transesophageal echocardiography demonstrated a round, echodense, mobile mass attached to the posterior mitral leaflet and annulus. At surgical exploration caseous annular calcification of the posterior mitral leaflet was diagnosed. After resection of the mass, successful mitral valvular plasty was performed. Review of the literature indicated that mitral annular calcification is associated with an increased risk of stroke. Optimal treatment may be surgery, especially when valve plasty can be performed, although randomized trials are currently lacking. PMID- 16378920 TI - Additive value of contrast echocardiography for the diagnosis of noncompaction of the left ventricular myocardium. AB - Noncompaction of the left ventricular myocardium is a rare and unclassified cardiomyopathy that remains frequently overlooked, even by experienced echocardiographers. This fact may be due to non-optimal imaging of the lateral and apical myocardium, and/or insufficient disease awareness by echocardiographers. We report a case of a young man with heart failure and severe left ventricular dysfunction, previously diagnosed of ischemic etiology, in which the contrast enhancement of left ventricular endocardium allowed us to reach the correct diagnosis of isolated left ventricular noncompaction. PMID- 16378921 TI - Neuropathic visceral dysmotility, brain cysts and calcifications, facial dysmorphism and developmental delay in two sibs. A new syndrome? AB - Syndromes with smooth muscle dysmotility are uncommon, and may be related either to smooth muscle myopathy, or to neuropathy. In most instances, neuropathic visceral dysmotility is an isolated finding leading to chronic intestinal pseudo obstruction syndrome (CIPO). We report here on two sibs, born to consanguineous parents, with neuropathic visceral dysmotility and CNS anomalies. They share facial dysmorphia, neurogenic megacystis, intracerebral calcifications, and developmental delay. The elder one, a girl, has microcephaly and multicystic kidneys, and her brother has a more extensive neuropathic visceral disorder leading clinically to CIPO. CIPO associated with megacystis is relatively frequent but is rarely associated with mental retardation. The cases reported in the literature are different from those described here, clinically and histologically. A recessively inherited form of CIPO associated with widespread intra-cerebral calcifications, malabsorption is known as Cockel syndrome. Severity of Cockel syndrome, absence of urinary tract involvement and neuropathologic discrepancies allow distinction with the disorder reported here. In conclusion, the two siblings described here have facial dysmorphia, vesical and (in one of them) intestinal neurogenic dysmotility, intracerebral calcifications and developmental delay that could represent a specific, recessively inherited form of CIPO. PMID- 16378922 TI - Novel GJA1 mutations in patients with oculo-dento-digital dysplasia (ODDD). AB - Oculo-dento-digital dysplasia (ODDD) is an autosomal dominant disorder characterized by developmental anomalies of the face, the eyes, the limbs and the teeth. Patients with ODDD usually present with complete syndactyly of the fourth and fifth fingers (type III syndactyly), ocular changes, abnormalities of primary and permanent dentition and specific craniofacial malformations. Mutations in GJA1, a gene that encodes the gap junction protein connexin 43, are responsible for ODDD. Gap junctions are assemblies of intercellular channels that allow exchange of various ions and signaling molecules between cells. In this way, gap junctions play an important regulatory role in a variety of physiologic and developmental processes. We identified three novel and one previously described GJA1 mutation in two large ODDD families and two sporadic ODDD cases. PMID- 16378923 TI - FISH screening for subtelomeric rearrangements in 219 patients with idiopathic mental retardation and normal karyotype. AB - Subtelomeric rearrangements are a common cause of idiopathic mental retardation (MR) accounting for 6.3-10.2% of moderate to severe cases and less than 1% of mildly retarded patients. We report on a cohort of 219 patients with idiopathic MR and normal 400-550 band karyotype screened for subtelomeric rearrangements by multiprobe Fluorescence in situ hybridization (FISH) in three Italian Genetics Centers. Twelve positive cases (5.5%) were found. Six were de novo deletions (1p, 7p, 9p, 9q, 20p, 22q) and four unbalanced translocations [a der(6)t(6q; 18p) and a der(18)t(8p; 18q) both de novo, a der(12)t(12p; 17q)mat and a der(2)t(2q; 17q) of unknown origin]. The remaining two cases were apparently balanced reciprocal translocations [a t(4p; 18q) and a t(1p; 16p)] of undetermined origin whose role in the pathogenesis of the clinical phenotype is doubtful. Dysmorphic features were present in all unbalanced patients, whilst a family history of MR was present in only four of them. The proposition that subtelomeric rearrangements are a significant cause of idiopathic MR is supported by our survey. Collection of the clinical data of positive patients will help to delineate the phenotype associated with the various subtelomeric abnormalities, to tailor healthcare services to the needs of these patients and their families and to determine the appropriate use of the test. PMID- 16378924 TI - Oculo-auriculo-vertebral spectrum (OAVS): clinical evaluation and severity scoring of 53 patients and proposal for a new classification. AB - Oculo-auriculo-vertebral spectrum (OMIM164210) is a phenotypically and probably also a genetically heterogeneous disorder, characterized by anomalies of the ear (mostly microtia), hemifacial microsomia, and defects of the vertebral column. Associated clinical findings include anomalies of the eye and brain, and developmental delay. We have evaluated the clinical data and photographs of 53 unrelated patients with OAVS, all presenting with either isolated microtia or preauricular tags in association with hemifacial microsomia as minimal diagnostic criteria; five had a positive family history for OAVS. Based on the main clinical findings and unilateral or bilateral involvement, we have developed a new classification system for OAVS, consisting of six subgroups. There is a statistically significant correlation between the subgroup and number of associated clinical findings, and a statistically significant difference regarding prognosis in uni- and bilaterally affected patients, suggesting that this classification is clinically relevant to the categorization of patients with OAVS. The newly developed scoring system (two points for each main clinical finding and one for each associated clinical finding) presented here, also aids prognosis, especially for delay of motor development and brain anomalies, and statistical analysis revealed significant clustering between different clinical findings of OAVS confirming the clinical impression previously published by several authors. PMID- 16378925 TI - Familial Mediterranean fever (FMF) in Lebanon and Jordan: a population genetics study and report of three novel mutations. AB - Familial Mediterranean fever (FMF) is an autosomal recessive disease mostly frequent in Mediterranean populations. Over 50 mutations have been identified in the gene responsible for the disease, MEFV. The present study reports the frequencies of MEFV mutations in 558 Lebanese and 55 Jordanian FMF patients and points out the severity of the M694V frequently observed mutation among these patients. Three novel mutations, T177I, S108R and E474K were also identified in the Lebanese group. An excess of homozygotes and a deficit of heterozygotes were observed in both samples when compared to the expected number of observed genotypes under the Hardy-Weinberg hypothesis. Homozygotes for M694V and M694I were still in excess in the Lebanese group of patients, even after consanguinous homozygotes were removed, or population structure was considered. This excess is therefore neither due to consanguinity nor to subgroups in the Lebanese population, but rather to more remote consanguinity or to a selection bias favoring the census of these genotypes. The fact that FMF female patients were less censed than male patients may be due to the greater resistance of females to pain and to the possibility of confusing abdominal and gynecological pain. The phenotypic heterogeneity of the FMF could then originate both from genetic causes like allelic heterogeneity or modulating genes, and cultural background facing the physiological consequences of genotypes at risk. PMID- 16378926 TI - Hyperechogenic fetal bowel: counseling difficulties. AB - The detection of echodense fetal bowel on ultrasound examination in the second trimester of pregnancy justifies invasive procedures such as amniocentesis to detect an underlying cause. We present a case in which initial tests identified only one mutation in the cystic fibrosis transmembrane regulator (CFTR)-gene of the fetus, the family history being negative for CF. Strongly reduced intestinal enzyme activities suggested intestinal obstruction and further increased the estimated risk for CF. After the 24th gestational week, a second mutation was found, confirming cystic fibrosis in this child. Problems in counseling in this particular case are discussed. PMID- 16378927 TI - 2nd Marie Curie Conference on arrayCGH and Molecular Cytogenetics (Project no. 504394-Project acronym arrayCGH-MC), (October 19-22), 2005, Bari, Italy. Abstracts. PMID- 16378928 TI - [Genetic diagnosis of Huntington disease]. AB - We report here a simple and effective method to assess the CAG repeat size of HD gene for gene diagnosis of Huntington disease. Genomic DNA sequences in polymorphic CAG repeat HD gene was amplified by PCR with TaKaRa LA Taq DNA polymerase and GC buffer. To distinguish normal alleles from HD alleles, DNA fragments of affected alleles were recovered as templates for secondary PCR. The secondary PCR products were cloned into T vector for sequencing to determine CAG repeat size. A total of 20 normal individuals and 3 members from a HD pedigree were included in this study. Results showed that the CAG repeat numbers in 20 normal individuals and 3 normal alleles from the HD pedigree varied in normal range, while in 3 HD alleles, the copy numbers of CAG repeat were 39, 40, 41 respectively. There was no overlap between the copy number of the normal and affected alleles. In conclusion, the TaKaRa LA Taq DNA polymerase with GC buffer can be used to effectively amplified CAG repeat of HD gene, which combined DNA sequencing can diagnose HD accurately. In addition, these finding suggested that the dynamic mutation in HD gene responsible for the genetic defect in Chinese HD patients. PMID- 16378929 TI - [Association study of NOTCH4 gene polymorphisms with schizophrenia and mood disorders in mixed pedigrees]. AB - This study was to explore the relationships between NOTCH4 gene and schizophrenia (SP) and mood disorders (MD), and to search for a common susceptible gene for SP and MD in Chinese Han population. We collected 61 mixed pedigrees of SP and MD in Chinese Han population. NOTCH4 polymorphisms -1725T/G and-25T/C were genotyped by applying PCR-RLFP technique, then transmission disequilibrium test (TDT) and haplotype-based haplotype relative risk analysis(HHRR) were performed. The results showed that -1725T/G was not associated with SP or MD (P>0.05), -25T/C was not associated with SP (P>0.05), but associated significantly with MD for female or early-onset (age of onset0.05). Our results suggested NOTCH4 or neighboring gene might be a common susceptible gene for SP and MD in the pedigrees studied. PMID- 16378930 TI - [Analysis of genetic polymorphism of 7 STR loci on chromosome 12 in Shaanxi Han populations]. AB - To analyze the genetic polymorphism of 7 STR loci (D12S1718,D12S1675, D12S358, D12S367, D12S1638, D12S1646 and D12S1682) on chromosome 12 in Shaanxi Hans. EDTA blood specimens were collected from 80 unrelated individuals from Chinese Han population in Shaanxi province. The DNA samples were extracted and relevant fragments were amplified by polymerase chain reaction (PCR). The PCR products were analyzed by ABI 3100 Genetic Analyzer. The number of alleles and genotypes observed at loci D12S1718, D12S1675, D12S358, D12S367, D12S1638, D12S1646 and D12S1682 were 7, 10, 8, 8, 6, 9, 11 for alleles and 10, 17, 18, 18, 14, 18, and 26 for genotypes, respectively. The heterozygosities for the 7 STR loci were 44.28%, 66.10%, 78.89%, 77.89%, 73.69%, 74.55% and 82.39%, respectively. The distribution of allele frequencies of 7 STR loci on chromosome 12 was consistent with Hardy-Weinberg equilibrium and relatively high genetic polymorphism was observed in Shaanxi Han population. PMID- 16378931 TI - [Study on the relationship between 22q11 microdeletion and congenital heart disease]. AB - To investigate if microdeletion of chromosome 22q11 is an epidemiologically important cause of congenital heart disease (CHD), we studied 25 cases with CHD phenotypes. Venous blood samples were tested by fluorescence in situ hybridization (FISH) for microdeletion of 22q11. Among 23 cases with simple CHD, 19 were shown not to have microdeletion of 22q11 and the other 4 cases were shown to have 22q11 microdeletion. Microdeletion of 22q11 was found in 2 cases with Tetralogy of Fallot (TOF) accompanied by multiple malformations. The results suggested that microdeletion of 22q11 was associated with CHD. PMID- 16378932 TI - [Study on centromeric dots variation of BGC823 cells and A549 cells]. AB - Chromosomal aneuploidy was a notable cytogenetic character in cancer cells. However, the mechanism of aneuploidy aberration is not clear up to the present. In order to probe into the mechanism of aneuploidy aberration of cancer cells, Centromeric dots (Cd) variation on chromosomes of BGC823 and A549 cells were studied by a simultaneous silver staining of both NOR and Cd from centromeric dots variation. The results showed that: (1) Frequency of Cd loss in BGC823 cells was 1.75%, Cd duplication laggard was 0.28%, small Cd was 1.82%, and Cd-NOR amalgamation was 0.95%. Compared to normal embryonic villi cells, frequencies of Cd loss and Cd-NOR amalgamation of BGC823 cells increased significantly (P<0.0125=, and that frequencies of Cd replication laggard and small Cd showed no difference between BGC823 cells and normal embryonic villi cells. (2) Frequency of Cd loss in A549 cells was 2.73%, Cd duplication laggard was 0.49%, small Cd was 1.73%, and Cd-NOR amalgamation was 0.71%. Frequencies of Cd loss and Cd replication laggard in A549 cells were significantly higher than those of normal embryonic villi cells(P<0.0125), moreover frequencies of Cd-NOR amalgamation and small Cd showed no difference between A549 cells and normal embryonic villi cells. We suggested that Cd loss and Cd-NOR amalgamation might be related with aneuploidy formation of BGC823 cells, and that Cd loss and Cd replication laggard might be related with aneuploidy formation of A549 cells. PMID- 16378933 TI - [Research on the polymorphism of heart fatty acid-binding protein gene in Shanxi pig breeds and their crossbred populations using PCR-RFLP]. AB - The genetic variation in 5' - upstream (Hinf I -RFLP)and the second intron (Hinf I *-RFLP, Hae III-RFLP)of heart fatty acid-binding protein(H-FABP)gene were detected with PCR-RFLP in 286 pigs including Mashen, Shanxi white pig and their crossbred populations. The results showed as follows: (1)Mashen presented only DD genotype while other populations varied,and Mashen crossbred populations had only 2 genotypes(DD, Dd) at the Hae III-RFLP site; (2)At the Hinf I -RFLP site of the 5' -upstream region, the crossbred population of Shanxi white pig and Duroc presented only HH genotype while other populations varied. Frequency of h allele in Mashen was 0.9727. (3)At the Hinf I *-RFLP site of the second intron, only Mashen presented 2 genotypes (BB, Bb), and frequency of B allele was 0.9667. (4)At the Hae III-RFLP and Hinf I *-RFLP sites, all populations were in Hardy weinberg equilibrium. PMID- 16378934 TI - [The genetic diversity and genetic effects of Myf-6 gene in different pig breeds]. AB - As a member of MRFs, Myf-6 gene plays an important role in the progress of myogenesis. The PCR-RFLP technique was applied in this study to analyze the distribution of Myf-6 gene in 12 different pig breeds and hybrid breeds. The genetic effects of Myf-6 gene on muscle fiber, carcass quality, carcass grading traits and meat quality traits were analyzed. The results indicated that: The polymorphisms of Myf-6/Ava I genotypes (intron1) were not abundant. Most of Chinese native pigs were AA homozygotes. B allele can be found only in foreign pig breeds and hybrid pigs originated from foreign pig breeds. Although no BB individuals were found in the experiment, the carcass lean percent of AB individuals (50.344 %) was significantly higher than that of AA individuals (45.875 %)(P<0.01), the loin eye area of AB individuals (27.097 cm2) was significantly larger than that of AA individuals (22.572 cm2)(P<0.01), and the carcass fat content of AB individuals (39.889 %) was significantly lower than that of AA individuals (44.503 %)(P<0.01). That means that the B allele can increase carcass lean percent and loin eye area, and decrease the carcass fat content, thus cause better carcass quality. No significant effects of the genotypes were found on the muscle fiber, FOM carcass grading traits and meat quality traits (P>0.05). PMID- 16378935 TI - [Influences of MyoG gene on reproductive traits in Jinhua pig]. AB - The genotypes in the second intron and 3' UTR region of myogenin gene in 149 Jinhua pigs (line I, 109; line II, 15; line III, 25) were detected by PCR-RFLP method. The results showed that three genotypes (AA, AB and BB) in the PCR1-MspI RFLP site were detected and their frequencies were 0.1544, 0.3826, 0.4631, respectively; MM and MN genotypes were found in PCR2-MspI-RFLP site and their frequencies were 0.9953 and 0.0047. The effect of MyoG gene on the reproductive traits was analyzed using SPSS software. Significant associations were found between MyoG locus and total number born (TNB) in first parity (P<0.05), and number born alive (NBA) in the second parity (P<0.05), but no significant associations existed in other parity combinations of TNB, NBA and litter weight birth (LWB) (P<0.05). PMID- 16378936 TI - [Genetic parameter estimation for inosine-5-monophosphate and intramuscular fat contents and other meat quality traits in chicken muscle]. AB - The genetic parameters for some important flavor traits like inosine-5' monophosphate (IMP) and intramuscular fat (IMF) contents in breast meat were estimated using a MTDFREML procedure on 1063 male, 90-day-old, purebred Beijing You meat-type chicks (BJY). The result showed that the heritability of IMP and IMF contents in BJY breast meat was moderate or low (h2=0.23, 0.10), whereas these parameters were higher for abdominal fat weight (AFW), breast meat yield (BMY), ratio of BMY to carcass weight (BMR), leg muscle yield (LMY), body weight (BW), comb weight(CW) and comb weight percentage (CWB) (h2=0.56-0.79). The heritability of abdominal fat percentage (AFP), leg meat yield (LMY), testicle weight (TW) and testicle weight percentage (TWP) were 0.24, 0.32, 0.39 and 0.35, respectively. IMP exhibited low phenotypic correlations with BMY, LMY and SFT and no significant phenotypic correlations with other traits. IMF, to some extent, exhibited positive phenotypic correlation with BW, AFP, SFT and FSW (rP=0.11 0.33). In terms of genetic correlation, IMP was moderately or significantly negatively correlated with BW and CWP (rA=-0.38,-0.62), and a high level of positive correlation was observed with BMY (rA=0.57). Moreover, IMF was highly correlated with BW and AFW (rA=0.75,0.66), and moderately correlated with AFP and CWP (rA=0.32, 0.40). A low level of positive correlation was observed between IMP and IMF (rA =0.27). We propose that IMP and IMF contents in chicken meat could be increased with selection through line-breeding. PMID- 16378937 TI - [Study on parentage testing in JIRONG rabbit by microsatellite markers]. AB - Thirteen microsatellite markers, including Sat2,Sat3,Sat4,Sat5,Sat7,Sat8,Sat12, Sat13, Sat16,Sol08,Sol28,Sol30 and Sol03, were studied for their parentage testing application in a group of 30 JIRONG Rabbits in the present report. The 13 microsatellite loci were successfully amplified with specific primers designed according to known sequences. The PCR products amplified from the microsatellite loci were analyzed by 8% denaturing polyacrylamide gel electrophoresis. The results demonstrated that the average alleles and the mean heterozygosity (Hs) and the polymorphism information content (PIC) and the combined exclusion probability (PE2) of the 13 microsatellite loci were 3.46, 0.578, 0.531, and 0.999329, respectively. The exclusion probability (PE1) of the 13 loci was 0.935226, and the confidence of the parentage testing was less than 80% when data of both parents were unknown, while the exclusion probability (PE1) and the confidence were 0.999329 and 95% respectively with known data of a single parent. Since the data of the rabbit maternal lines studied were known, the paternal lines of the group were successfully identified using the 13 microsatellite loci with high confidence. PMID- 16378938 TI - [Molecular cloning of full-long cDNA sequences encoding hairless gene in the Kunming mouse]. AB - The mouse hairless gene is a crucial nuclear receptor gene associated with the structure of hair and skin. It encodes a putative zinc finger transcription factor, and is a transcriptional corepressor for thyroid hormone receptors. Hairless gene plays a critical role in maintaining the delicate balance between cell proliferation, differentiation, and apoptosis in the hair follicle as well as in the interfollicular epidermis, and is concerned with the control of hair growth cycling. This study was designed to clone and analyze the cDNA encoding Hr from Kunming mouse. The RT-PCR method was developed to clone the Hr cDNA. A full length cDNA and CDS sequences of Kunming mouse were 4104bp and 3546bp, respectively, which has been accepted by GenBank (Accession Number: AY547391). Accession number of protein encoded by AY547391 sequence in GenBank is AAT45233 , composed of 1181 amino acid residues. The Hr cDNA of Kunming mouse was cloned and sequenced for the first time. The identities of CDS of Hr gene were 99.9%, 94.4%, 83.1%, 78.1%, 81.9% and 82.1% by homologous comparison among Kunming mouse and other six species, and that were 99.9%, 92.2%, 81.7%, 70.8%, 79.9% and 80.1% respectively in amino acid sequences. The results suggested a high degree of conservation and thus functional significance of the Hr gene among different mammalian taxons. The results derived from information searching by Blast program revealed that there were 4 SNP sites and one deletion mutation in the sequences of hairless gene mRNAs between Kunming mice and that collected in GenBank. Three SNP sites did not alter the related amino acids encoded. Two SNP proved to be polymorphism mutant sites with amino acids changes. The results provided new data for SNPs in Hr gene. PMID- 16378939 TI - [Research on the genetic polymorphism in crested ibis by RMAPD]. AB - The genetic polymorphism of Yangxian artificially reared 43 crested ibises (Nipponia nippon) was firstly investigated by RMAPD (random microsatellite amplify polymorphic DNA) marker. The results showed that RMAPD was more stable and polymorphic than RAPD. 2147 bands were amplified by 12 pairs of primers,of which 93 bands had polymorphisms. The band sharing frequency and the genetic diversity index were 0.718 and 3.664, respectively, indicating that the genetic structure of crested ibis population was simple and poor genetic variations existed in crested ibis population in Yangxian. It is imperious to protect the genetic diversity of crested ibis population. PMID- 16378940 TI - [A preliminary study on the inheritance of microsatellite in two selective breeding families of shrimp (Litopenaeus vannamei)]. AB - Two full-sib families of Litopenaeus vannamei were used to study the inheritance of allelic variation at 10 microsatellite loci. Of the 20 genotypic ratios observed (10 loci x 2 crosses), 17 ratios conformed to Mendelian segregation. When null alleles were considered, one loci (TUMXLv8.220) confirmed Mendelian expectations in all families. While one loci (TUMXLv3.1) showed deviation from family 06. Three loci (TUMXLv5.66,TUMXLv7.74,TUMXLv8.224)were monomorphic in both controlled crosses; 3 loci were polymorphic and confirmed to Mendelian ratios in all families, can be used for parental analysis and population genetic studies. These results indicated the need to test the inheritance pattern for microsatellite markers in shrimp before using them for population genetic or kinship analysis. PMID- 16378941 TI - [Study on the genetic basis of plant height and ear height in maize (Zea mays L.) by QTL dissection]. AB - One hundred and ninety-one F2 individuals derives from the cross, Mo17xHuangzao4, were genotyped by SSR and AFLP markers to construct the genetic linkage map, and 184 corresponding F2:3 families were phenotyped for maize (Zea mays L.) plant hright and ear height in Changping and Shunyi. A mixed linear model approach and its software were used to detect QTLs with main effect, QTLs involved in digenic interations and Q x E interations. In total, 7 QTLs of plant height, 18 pairs of digenic epistatic loci of plant height, 13 pairs of digenic epistatic loci of ear height were detected. It was found that 6 QTLs of plant height, 8 QTLs of ear height, and 4 pairs of digenic epistatic loci of plant height and ear height and a significant interaction with environments. Genetic compnents underlying these quantitative traits were analyzed, and the study showed that additive, dominance and epistaship between plant height and ear height, and the results described above were also discussed for their possible application in molecular breeding. PMID- 16378942 TI - [The effect of phKL gene on homoeologous pairing of wheat-alien hybrids is situated between gene mutants of Ph1 and Ph2]. AB - In natural populations of common wheat landrace, there has a phKL gene promoting homoeologous pairing of wheat-alien hybrids. In this study, the effects were compared among phKL, ph1b, ph2a and ph2b on homoeologous pairing of wheat-alien hybrids. The effects were indicated as ph1b > phKL > ph2b > ph2a, i. e. phKL gene was situated between gene mutants of Ph1 and Ph2. PMID- 16378943 TI - [Isolation and sequence analysis of a omega-gliadin homologous gene from wheat]. AB - The DNA sequence of a full-length Triticum astivum CV. Jinan 177omega-gliadin homologous gene (omega1236) containing partial 5' and 3' flanking sequences with no intron was cloned by genomic PCR-based technology. The omega1236 sequence possibly encode a putative 47.2 kDa protein except for eight stop codons at amino acid residue positions 87, 117, 125, 157, 198, 313, 357 and 365 respectively. All the eight stop codons were caused by base transition. Sequence analysis revealed that omega1236 had 98% homology to a omega-gliadin gene of wheat (AB059812). Like all other gliadin gene families characterized in cereals, this gene possessed all the features in other plant reported previously. Phylogenetic analysis of the completely sequenced gene as well as those omega-genes in wheat, omega-secalin and C-horden genes in rye and barley, and alpha-, beta- and gamma-gliadin genes in wheat indicated that the omega1236 was more closely related to omega-gliadin gene family, much less homology to alpha- , beta- and gamma-gliadin gene families. Short peptide was produced in the culture of transformed E. coli induced by IPTG in early 2 h. It indicated that stop codon would be in omega1236. The result is consistent with that of the sequenced gene. The present paper could accumulate data useful for both omega-gliadin gene cloning by PCR and the study on structures and functions of these genes. PMID- 16378944 TI - [The heredity of flower colors and the discovery of flower color chimera in chrysanthemum species]. AB - The reciprocal crosses of yellow colored chrysanthemum x red colored chrysanthemum and white colored chrysanthemum x red colored chrysanthemum were conducted in order to analyze the heredity of flower colors. The results revealed that the heredity of flower colors was very complicated, and mainly exhibited matroclinous characteristics when red colored materials was used as maternal parent but not in the combinations when the yellow or white colored materials were used as maternal parents. The incomplete dominance and mosaic dominance also existed in the heredity of chrysanthemum flower colors. The flower-color chimeras with two kinds of flower buds were discovered in the cross of 3501 x 3509, i.e. one side of the flower buds was completely in red color, which was same as the parental material of 3509, and another side was generally in yellow color with red spots on them. Cytological analysis showed that two sides were both with 36 chromosomes, indicating that the formation of chimera was not resulted from the changes of chromosome numbers, but from the destruction of pigment synthesis genes by the insert of transposable element. PMID- 16378945 TI - [The application of Red/ET recombination to high efficient gene-targeting vector construction]. AB - A rapid and high efficient working system for gene-targeting vector construction was developed by using Red/ET recombination. Mediated by Red/ET recombination, the objective genomic DNA was first subcloned into the targeting vector. After insertion of a PCR amplified selectable marker gene flanked with short homology arms into the targeted position, a conventional gene knock-out targeting vector was then constructed. For conditional gene knock-out targeting vector construction, with the co-operation of Cre-loxP site-specific recombination, two rounds of Red/ET recombination was just needed. Being different from PCR and endonuclease-based gene-targeting vector construction, the homologous regions used for gene targeting can be chosen as long as possible. Furthermore, no enzyme digestion, ligation and sequencing identification were involved, so that it is very efficient and labor-saving. Several different gene-targeting vectors were successfully constructed by using this system. The establishment of this working system will accelerate the gene function studies in the post-genome stage. PMID- 16378946 TI - [Analysis of the coding region for signal peptide-containing proteins in Pseudomonas syringae pv. tomato genome]. AB - The completed 5 615 ORFs of chromosome sequence of Pseudomonas syringae pv. tomato were analyzed by SignalP 3.0. The result revealed that 679 ORFs consisted of putative secretary proteins with signal peptides. 107 ORFs of signal peptides have been named. The length of most signal peptides was different from 19 amino acids to 31 amino acids, and the most dominant one was 23 amino acids in length. The size of most ORFs varied between 101 ~ 400 amino acids. The frequency of particular amino acids in signal peptides by statistical approach was 48.54% for hydrophobic, 18.67% for uncharged, 24.54% for negative and 8.00% for positive amino acids. The most frequent amino acid was alanine, and the least was isoleucine. Three classes of signal peptides were found in the genome of P. syringae pv. tomato: 501 ORFs belong to secretary signal peptides, 36 ORFs belong to twin-arginine signal peptides, and 15 ORFs belong to lipoprotein signal peptides. Type IV pilin signal peptide and bacteriocins and pheronoes signal peptide was not found in the genome. PMID- 16378947 TI - [Study on PBPs genes of penicillin-resistant Streptococcus pneumonia in Zhejiang province]. AB - To elucidate alternations in gene/amino acid sequence of penicillin-binding proteins (PBPs) 1A, 2B, 2X from clinical isolates of penicillin-resistant Streptococcus pneumonia (PRSP) in Zhejiang Province, 26 strains of Streptococcus pneumonia were collected from November 2001 to January 2004. The antibiotics susceptibility of these strains was detected. PCR amplification and direct sequencing of PBP1A, 2B, 2X genes were performed. The sequence variations of PBP genes of the PRSPs in this region were studied by sequence BLAST analysis. It was shown that the main alternations of PBP1A were the four consecutive amino acid substitutions (Thr574Ala, Ser575Thr, Gln576Gly, Phe577Tyr) following the conservative motif KTG and the amino acid substitution Thr371Ser in the conservative motif STMK. The main alternation of PBP2B was Thr451Ala following the conservative motif SSN, and the main alternation of PBP2X was Thr338Ala in conservative motif STMK. The above mutation sites and drug resistant level were consistent to the data reported previously. Neither new gene mutation specific to these strains nor certain amino acid substitutions related to penicillin resistance reported was identified in the genes. PMID- 16378949 TI - [Improvement and optimization for "genetics" course teaching]. AB - "Genetics" is one of the main courses for the study scheme of undergraduate students in university. This paper pointed out some problems found in the teaching process according to the characteristics of "Genetics" course. At the same time, the thought on the construction of "Genetics" course were discussed concerning the reform of course contents, the improvement of teaching method, the creation of teaching CAI (computer assisted instruction), the compile of electronic edition for teaching materials, the evaluation of teaching level, the innovation of experimental contents and the methods for improving the teaching effect. PMID- 16378948 TI - [The "horse-shoe effect"of correspondence analysis for human population genetic structure and its population genetic explanation]. AB - At present study, the reasons of "horse-shoe effect" in correspondence analysis for analyzing human population genetic structure was explained. Based on the structure of gene frequency matrix, we displaye the different patterns of Scallergram of correspondent analysis from different types of loci (HLA-A locus, and STR- CSF1PO locus in Chinese Han populations). The results indicate that different types of loci showed different patterns of Scallergram of correspondent analysis. When some alleles have very low frequency in the gene frequency matrix, there would be "horse-shoe effect" in the Scallergram of correspondent analysis. The reason is that the c2 distance measurement in correspondent analysis usually overrates the effect of the genes with low frequencies. To carry out the correspondent analysis of human population genetic structure, when the Scallergram presents "horse-shoe effect", one should examine the structure of gene frequency matrix, and confirm whether the "horse-shoe effect" shows the real pattern of population genetic structure. Only in this way, one can explain the "horse-shoe effect" correctly. PMID- 16378950 TI - [Mutations and polymorphisms of the P gene associated with oculocutaneous albinism type II]. AB - Oculocutaneous albinism typeII(OCA2), the most common type of albinism, is an autosomal recessive disorder. It is caused by mutations in the P gene, which is located on chromosome 15q11.1-q12 and divided into 24 exons and 23 introns. P gene codes for 838-amino-acid integral membrane protein with 12 putative transmembrane domains, but the exact function is not clear yet. There are at least 60 pathologic mutations and 43 non-pathologic polymorphisms have been found. Pathologic mutations include missense mutations, nonsense mutations, frameshift mutations and splice-sit mutations. But unlike TYR gene, most of P gene mutations are located on the C-terminal and don' t cluster in defined regions. It is difficult to define the pathologic mutations since many non pathologic polymorphisms also lie in exons. Some non-pathologic missense mutations may be associated with phenotypic variation in normally pigmented individuals and need to further study. PMID- 16378951 TI - [Hira gene and development: from yeast to human]. AB - HIR/HIRA, one of the histone chaperones, encoded by Hir/Hira (histone regulation) genes, were initially identified in yeast as negative regulators of histone gene expression. It has been confirmed that HIRA contains a conserved family of proteins found in various species including low eukaryotes, invertebrates and vertebrates. It is essential for proper development. Mutations of Hir/Hira genes result in very serious defects in normal development not only in yeast but also in advanced eukaryotes. Combined with the work in our group, the roles of Hir/Hira genes in the development in different species are reviewed, and the recent advances concerning the mechanisms of HIRA functioning are also summarized in an attempt to promote the research further on the exact roles of hira genes in development of eukaryotes and on their deeply operating mechanisms. PMID- 16378952 TI - [Research progress in human artificial chromosomes(HACs) and the potentials in application]. AB - Since the first report of the establishment of human artificial chromosome(HAC) was published in 1997, several types of HAC have been created by different strategies. Compared to other artificial chromosomes, such as yeast artificial chromosome (YAC) and bacterial artificial chromosome(BAC), HAC exists in a cell independently, in other words, HAC does not integrated into the cellular genome, and can undergo normal mitosis and meiosis from generation to generation in vitro and in vivo. Recent results proved that HAC, as a DNA carrier, is able to host a large fragment of DNA or mini-chromosome, thus it could be a very important tool in the study of human gene expression and regulation, human chromosome function and minimum functional elements and animal models for human diseases. In the near future, HAC can also be used in gene therapy for human genetic diseases. PMID- 16378953 TI - [The application of bioinformatics in the research of alternative splicing]. AB - Alternative splicing, a fundamental and important regulatory mechanism in eukaryotes, allows one pre-mRNA to be processed into many different mature forms within a cell, each of which can have distinct functions. As alternative splicing is associated with human diseases, the study of alternative splicing becomes quite important. Bioinformatics is a new subject for the study of alternative splicing, especially for its regulatory mechanism, prediction and origin. Of course, bioinformatics must be combined with experimental research so as to clarify these aspects of alternative splicing. This paper reviewed the recent research progress in this field in the hope to gain a deeper understanding of eukaryotic gene expression regulation. PMID- 16378954 TI - [Gene cloning and structure characterization of CMS/Rf system in plants]. AB - It has achieved great success in plant heterosis by using cytoplasmic male sterility and fertility restorer (CMS/Rf) system. In order to elucidate the action mechanism of Rf gene in this system, many researchers have been contributing themselves to map and clone fertility restorer genes in recent years. Four Rf genes were recently cloned successfully. Here we presented the overview of plant CMS fertility restorer gene mapping, cloning and its molecular mechanism. It was also discussed on utilization of fertility restorer genes in the molecular breeding in plants. PMID- 16378955 TI - [Progress in regulation of rice Wx gene expression]. AB - The granule-bound starch synthase(GBSS) is encoded by the rice Wx gene, which is the major gene for control of amylase synthesis of. This paper briefly introduced the progress in regulation of rice Wx gene expression at transcriptional level and post-transcriptional level. At the same time the influence of transgene, genetic background and temperature condition on Wx gene expression in rice was discussed. Finally some urgent questions were suggested for the further study on regulation of rice Wx gene expression. PMID- 16378956 TI - [Molecular evidence for origin of plastid-like organelle apicoplast: a review]. AB - Apicomplexan protozoa contains a highly reduced plastid-like organelle termed apicoplast. The research on evolutionary origin of apicoplast is a topic of long term intense debate, by far, hasn't still reached congruent conclusion though a variety of molecular technologies were employed, so that it become a typical case in plastid origin field. In this paper , we reviewed the molecular evidence for apicoplast evolutionary origin, predicting the potential source of novel molecular evidence as clues for further studying. PMID- 16378957 TI - Characterization of a vaccinia virus mutant with a deletion of the D10R gene encoding a putative negative regulator of gene expression. AB - The D9 and D10 proteins of vaccinia virus are 25% identical to each other, contain a mutT motif characteristic of nudix hydrolases, and are conserved in all sequenced poxviruses. Previous studies indicated that overexpression of D10 and, to a lesser extent, D9 decreased the levels of capped mRNAs and their translation products. Here, we further characterized the D10 protein and showed that only trace amounts are associated with purified virions and that it is expressed exclusively at late times after vaccinia virus infection. A viable deletion mutant (vdeltaD10) produced smaller plaques and lower virus yields than either wild-type virus or a D9R deletion mutant (vdeltaD9). Purified vdeltaD10 virions appeared normal by microscopic examination and biochemical analysis but produced 6- to 10-fold-fewer plaques at the same concentration as wild-type or vdeltaD9 virions. When 4 PFU per cell of wild-type or vdeltaD9 virions or equal numbers of vdeltaD10 virions were used for inoculation, nearly all cells were infected in each case, but viral early and late transcription was initiated more slowly in vdeltaD10-infected cells than in the others. However, viral early transcripts accumulated to higher levels in vdeltaD10-infected cells than in cells infected with the wild type or vdeltaD9. In addition, viral early and late mRNAs and cellular actin mRNA persisted longer in vdeltaD10-infected cells than in others. Furthermore, analysis of pulse-labeled proteins indicated prolonged synthesis of cellular and viral early proteins. These results are consistent with a role for D10 in regulating RNA levels in poxvirus-infected cells. PMID- 16378958 TI - Evolution of broad host range in retroviruses leads to cell death mediated by highly cytopathic variants. AB - The ability of many retroviruses to cause disease can be correlated to their cytopathic effect (CPE) in tissue culture characterized by an acute period of cell death and viral DNA accumulation. Here, we show that mutants of a subgroup B avian retrovirus (Alpharetrovirus) cause a very dramatic CPE in certain susceptible avian cells that is coincident with elevated levels of apoptosis, as measured by nuclear morphology, and persistent viral DNA accumulation. These mutants also have a broadly extended host range that includes rodent, cat, dog, monkey, and human cells (31). Previously, we have shown that the mutants exhibit diminished resistance to superinfection. The results presented here have important implications for the process of evolution of retroviruses to use distinct cellular receptors. PMID- 16378959 TI - Sequence homology required by human immunodeficiency virus type 1 to escape from short interfering RNAs. AB - Short interfering RNAs (siRNAs) targeting viral or cellular genes can efficiently inhibit human immunodeficiency virus type 1 (HIV-1) replication. Nevertheless, the emergence of mutations in the gene being targeted could lead to the rapid escape from the siRNA. Here, we simulate viral escape by systematically introducing single-nucleotide substitutions in all 19 HIV-1 residues targeted by an effective siRNA. We found that all mutant viruses that were tested replicated better in the presence of the siRNA than in the presence of the wild-type virus. The antiviral activity of the siRNA was completely abolished by single substitutions in 10 (positions 4 to 11, 14, and 15) out of 16 positions tested (substitution at 3 of the 19 positions explored rendered nonviable viruses). With the exception of the substitution observed at position 12, substitutions at either the 5' end or the 3' end (positions 1 to 3, 16, and 18) were better tolerated by the RNA interference machinery and only in part affected siRNA inhibition. Our results show that optimal HIV-1 gene silencing by siRNA requires a complete homology within most of the target sequence and that substitutions at only a few positions at the 5' and 3' ends are partially tolerated. PMID- 16378960 TI - The MC160 protein expressed by the dermatotropic poxvirus molluscum contagiosum virus prevents tumor necrosis factor alpha-induced NF-kappaB activation via inhibition of I kappa kinase complex formation. AB - The pluripotent cytokine tumor necrosis factor alpha (TNF-alpha) binds to its cognate TNF receptor I (TNF-RI) to stimulate inflammation via activation of the NF-kappaB transcription factor. To prevent the detrimental effects of TNF-alpha in keratinocytes infected with the molluscum contagiosum virus (MCV), this poxvirus is expected to produce proteins that block at least one step of the TNF RI signal transduction pathway. One such product, the MC160 protein, is predicted to interfere with this cellular response because of its homology to other proteins that regulate TNF-RI-mediated signaling. We report here that expression of MC160 molecules did significantly reduce TNF-alpha-mediated NF-kappaB activation in 293T cells, as measured by gene reporter and gel mobility shift assays. Since we observed that MC160 decreased other NF-kappaB activation pathways, namely those activated by receptor-interacting protein, TNF receptor associated factor 2, NF-kappaB-inducing kinase, or MyD88, we hypothesized that the MC160 product interfered with I kappa kinase (IKK) activation, an event common to multiple signal transduction pathways. Indeed, MC160 protein expression was associated with a reduction in in vitro IKK kinase activity and IKK subunit phosphorylation. Further, IKK1-IKK2 interactions were not detected in MC160 expressing cells, under conditions demonstrated to induce IKK complex formation, but interactions between the MC160 protein and the major IKK subunits were undetectable. Surprisingly, MC160 expression correlated with a decrease in IKK1, but not IKK2 levels, suggesting a mechanism for MC160 disruption of IKK1-IKK2 interactions. MCV has probably retained its MC160 gene to inhibit NF-kappaB activation by interfering with signaling via multiple biological mediators. In the context of an MCV infection in vivo, MC160 protein expression may dampen the cellular production of proinflammatory molecules and enhance persistent infections in host keratinocytes. PMID- 16378961 TI - Point mutations upstream of hepatitis B virus core gene affect DNA replication at the step of core protein expression. AB - The pregenomic RNA directs replication of the hepatitis B virus (HBV) genome by serving both as the messenger for core protein and polymerase and as the genome precursor following its packaging into the core particle. RNA packaging is mediated by a stem-loop structure present at its 5' end designated the epsilon signal, which includes the core gene initiator AUG. The precore RNA has a slightly extended 5' end to cover the entire precore region and, consequently, directs the translation of a precore/core protein, which is secreted as e antigen (HBeAg) following removal of precore-derived signal peptide and the carboxyl terminus. A naturally occurring G1862T mutation upstream of the core AUG affects the bulge of the epsilon signal and generates a "forbidden" residue at the -3 position of the signal peptide cleavage site. Transfection of this and other mutants into human hepatoma cells failed to prove their inhibition of HBeAg secretion but rather revealed great impairment of genome replication. This replication defect was associated with reduced expression of core protein and could be overcome by a G1899A covariation, or by nonsense or frameshift mutation in the precore region. All these mutations antagonized the G1862T mutation on core protein expression. Cotransfection of the G1862T mutant with a replication deficient HBV genome that provides core protein in trans also restored genome replication. Consistent with our findings in cell culture, HBV genotype A found in African/Asian patients has T1862 and is associated with much lower viremia titers than the European subgroup of genotype A. PMID- 16378962 TI - Inhibition of protease-resistant prion protein formation in a transformed deer cell line infected with chronic wasting disease. AB - Chronic wasting disease (CWD) is an emerging transmissible spongiform encephalopathy (prion disease) of North American cervids, i.e., mule deer, white tailed deer, and elk (wapiti). To facilitate in vitro studies of CWD, we have developed a transformed deer cell line that is persistently infected with CWD. Primary cultures derived from uninfected mule deer brain tissue were transformed by transfection with a plasmid containing the simian virus 40 genome. A transformed cell line (MDB) was exposed to microsomes prepared from the brainstem of a CWD-affected mule deer. CWD-associated, protease-resistant prion protein (PrP(CWD)) was used as an indicator of CWD infection. Although no PrP(CWD) was detected in any of these cultures after two passes, dilution cloning of cells yielded one PrP(CWD)-positive clone out of 51. This clone, designated MDB(CWD), has maintained stable PrP(CWD) production through 32 serial passes thus far. A second round of dilution cloning yielded 20 PrP(CWD)-positive subclones out of 30, one of which was designated MDB(CWD2). The MDB(CWD2) cell line was positive for fibronectin and negative for microtubule-associated protein 2 (a neuronal marker) and glial fibrillary acidic protein (an activated astrocyte marker), consistent with derivation from brain fibroblasts (e.g., meningeal fibroblasts). Two inhibitors of rodent scrapie protease-resistant PrP accumulation, pentosan polysulfate and a porphyrin compound, indium (III) meso-tetra(4 sulfonatophenyl)porphine chloride, potently blocked PrP(CWD) accumulation in MDB(CWD) cells. This demonstrates the utility of these cells in a rapid in vitro screening assay for PrP(CWD) inhibitors and suggests that these compounds have potential to be active against CWD in vivo. PMID- 16378963 TI - Restriction of foamy viruses by APOBEC cytidine deaminases. AB - Foamy viruses (FVs) are nonpathogenic retroviruses infecting many species of mammals, notably primates, cattle, and cats. We have examined whether members of the apolipoprotein B-editing catalytic polypeptide-like subunit (APOBEC) family of antiviral cytidine deaminases restrict replication of simian FV. We show that human APOBEC3G is a potent inhibitor of FV infectivity in cell culture experiments. This antiviral activity is associated with cytidine editing of the viral genome. Both molecular FV clones and primary uncloned viruses were susceptible to APOBEC3G, and viral infectivity was also inhibited by murine and simian APOBEC3G homologues, as well as by human APOBEC3F. Wild-type and bet deleted viruses were similarly sensitive to this antiviral activity, suggesting that Bet does not significantly counteract APOBEC proteins. Moreover, we did not detect FV sequences that may have been targeted by APOBEC in naturally infected macaques, but we observed a few G-to-A substitutions in humans that have been accidentally contaminated by simian FV. In infected hosts, the persistence strategy employed by FV might be based on low levels of replication, as well as avoidance of cells expressing large amounts of active cytidine deaminases. PMID- 16378964 TI - Morphological changes in the T=3 capsid of Flock House virus during cell entry. AB - We report the identification and characterization of a viral intermediate formed during infection of Drosophila cells with the nodavirus Flock House virus (FHV). We observed that even at a very low multiplicity of infection, only 70% of the input virus stayed attached to or entered the cells, while the remaining 30% of the virus eluted from cells after initial binding. The eluted FHV particles did not rebind to Drosophila cells and, thus, could no longer initiate infection by the receptor-mediated entry pathway. FHV virus-like particles with the same capsid composition as native FHV but containing cellular RNA also exhibited formation of eluted particles when incubated with the cells. A maturation cleavage-defective mutant of FHV, however, did not. Compared to naive FHV particles, i.e., particles that had never been incubated with cells, eluted particles showed an acid-sensitive phenotype and morphological alterations. Furthermore, eluted particles had lost a fraction of the internally located capsid protein gamma. Based on these results, we hypothesize that FHV eluted particles represent an infection intermediate analogous to eluted particles observed for members of the family Picornaviridae. PMID- 16378965 TI - Addition of N-glycans in the stalk of the Newcastle disease virus HN protein blocks its interaction with the F protein and prevents fusion. AB - Most paramyxovirus fusion (F) proteins require the coexpression of the homologous attachment (HN) protein to promote membrane fusion, consistent with the existence of a virus-specific interaction between the two proteins. Analysis of the fusion activities of chimeric HN proteins indicates that the stalk region of the HN spike determines its F protein specificity, and analysis of a panel of site directed mutants indicates that the F-interactive site resides in this region. Here, we use the addition of oligosaccharides to further explore the role of the HN stalk in the interaction with F. N-glycans were individually added at several positions in the stalk to determine their effects on the activities of HN, as well as its structure. N-glycan addition at positions 69 and 77 in the stalk specifically blocks fusion and the HN-F interaction without affecting either HN structure or its other activities. N-glycans added at other positions in the stalk modulate activities that reside in the globular head of HN. This correlates with an alteration of the tetrameric structure of the protein, as indicated by sucrose gradient sedimentation analyses. Finally, N-glycan addition in another region of HN (residues 124 to 152), predicted by a peptide-based analysis to mediate the interaction with F, does not significantly reduce the level of fusion, arguing strongly against this site being part of the F-interactive domain in HN. Our data support the idea that the F-interactive site on HN is defined by the stalk region of the protein. PMID- 16378966 TI - Perturbations of cell cycle control in T cells contribute to the different outcomes of simian immunodeficiency virus infection in rhesus macaques and sooty mangabeys. AB - In contrast to human immunodeficiency virus (HIV) infection of humans and experimental simian immunodeficiency virus (SIV) infection of rhesus macaques (RMs), SIV infection of sooty mangabeys (SMs), a natural host African monkey species, is typically nonpathogenic and associated with preservation of CD4+ T cell counts despite chronic high levels of viral replication. In previous studies, we have shown that the lack of SIV disease progression in SMs is related to lower levels of immune activation and bystander T-cell apoptosis compared to those of pathogenic HIV/SIV infection (G. Silvestri, D. Sodora, R. Koup, M. Paiardini, S. O'Neil, H. M. McClure, S. I. Staprans, and M. B. Feinberg, Immunity 18:441-452, 2003; G. Silvestri, A. Fedanov, S. Germon, N. Kozyr, W. J. Kaiser, D. A. Garber, H. M. McClure, M. B. Feinberg, and S. I. Staprans, J. Virol. 79:4043 4054, 2005). In HIV-infected patients, increased T-cell susceptibility to apoptosis is associated with a complex cell cycle dysregulation (CCD) that involves increased activation of the cyclin B/p34-cdc2 complex and abnormal nucleolar structure with dysregulation of nucleolin turnover. Here we report that CCD is also present during pathogenic SIV infection of RMs, and its extent correlates with the level of immune activation and T-cell apoptosis. In marked contrast, naturally SIV-infected SMs show normal regulation of cell cycle control (i.e., normal intracellular levels of cyclin B and preserved nucleolin turnover) and a low propensity to apoptosis in both peripheral blood- and lymph node derived T cells. The absence of significant CCD in the AIDS-free, non-immune activated SMs despite high levels of viral replication indicates that CCD is a marker of disease progression during lentiviral infection and supports the hypothesis that the preservation of cell cycle control may help to confer the disease-resistant phenotype of SIV-infected SMs. PMID- 16378968 TI - The transcription profile of Aleutian mink disease virus in CRFK cells is generated by alternative processing of pre-mRNAs produced from a single promoter. AB - A reevaluation of the transcription profile of Aleutian mink disease parvovirus (AMDV)-infected CRFK cells at either 32 degrees C or 37 degrees C has determined that strain AMDV-G encodes six species of mRNAs produced by alternative splicing and alternative polyadenylation of a pre-mRNA generated by a single promoter at the left end of the genome. Three different splicing patterns are used, and each type is found polyadenylated at either the 3' end of the genome (the distal site) or at a site in the center of the genome (the proximal site). All spliced species accumulate similarly over the course of infection, with the R2 RNA predominant throughout. The R2 RNA, which contains and can express the NS2 coding region, encodes the viral capsid proteins VP1 and VP2. PMID- 16378967 TI - Dynamics of hepatitis B virus resistance to lamivudine. AB - Lamivudine was the first approved inhibitor of hepatitis B virus (HBV) reverse transcriptase (RT). Lamivudine resistance develops in 53% to 76% of patients after 3 years of treatment. We extensively characterized the dynamics of HBV quasispecies variant populations in four HBV-infected patients who developed lamivudine resistance. Virological breakthrough was preceded by 2 to 4 months by the emergence of quasispecies variants bearing amino acid substitutions at RT position 204, i.e., within the YMDD catalytic motif (rtM204V/I). Three patients had a gradual switch from a YMDD variant population at baseline to a 100% lamivudine-resistant variant population, whereas the remaining patient had a fluctuating pattern of resistance variant dynamics. Careful analysis of amino acid substitutions located outside domain C of HBV RT, including those known to partially restore replication capacities in vitro, showed that the in vivo replication of HBV variants is driven by multiple forces, including intrinsic replicative advantages conferred by mutations accumulating outside domain C and the changing environment in which these variants replicate. Our findings also suggest that individual treatment optimization will require sensitive methods capable of detecting the emergence of viral resistance before the relevant variants acquire optimal replicative capacities. PMID- 16378969 TI - Expanded tissue targets for foamy virus replication with simian immunodeficiency virus-induced immunosuppression. AB - Foamy viruses (FV) are the oldest known genus of retroviruses and have persisted in nonhuman primates for over 60 million years. FV are efficiently transmitted, leading to a lifelong nonpathogenic infection. Transmission is thought to occur through saliva, but the detailed mechanism is unknown. Interestingly, this persistent infection contrasts with the rapid cytopathicity caused by FV in vitro, suggesting a host defense against FV. To better understand the tissue specificity of FV replication and host immunologic defense against FV cytopathicity, we quantified FV in tissues of healthy rhesus macaques (RM) and those severely immunosuppressed by simian immunodeficiency virus (SIV). Contrary to earlier findings, we find that all immunocompetent animals consistently have high levels of viral RNA in oral tissues but not in other tissues examined, including the small intestine. Strikingly, abundant viral transcripts were detected in the small intestine of all of the SIV-infected RM, which has been shown to be a major site of SIV (and human immunodeficiency virus)-induced CD4+ T cell depletion. In contrast, there was a trend to lower viral RNA levels in oropharyngeal tissues of SIV-infected animals. The expansion of FV replication to the small intestine but not to other CD4+ T-cell-depleted tissues suggests that factors other than T-cell depletion, such as dysregulation of the jejunal microenvironment after SIV infection, likely account for the expanded tissue tropism of FV replication. PMID- 16378970 TI - Reovirus variants selected for resistance to ammonium chloride have mutations in viral outer-capsid protein sigma3. AB - Mammalian reoviruses are internalized into cells by receptor-mediated endocytosis. Within the endocytic compartment, the viral outer capsid undergoes acid-dependent proteolysis resulting in removal of the sigma3 protein and proteolytic cleavage of the mu1/mu1C protein. Ammonium chloride (AC) is a weak base that blocks disassembly of reovirus virions by inhibiting acidification of intracellular vacuoles. To identify domains in reovirus proteins that influence pH-sensitive steps in viral disassembly, we adapted strain type 3 Dearing (T3D) to growth in murine L929 cells treated with AC. In comparison to wild-type (wt) T3D, AC-adapted (ACA-D) variant viruses exhibited increased yields in AC-treated cells. AC resistance of reassortant viruses generated from a cross of wt type 1 Lang and ACA-D variant ACA-D1 segregated with the sigma3-encoding S4 gene. The deduced sigma3 amino acid sequences of six independently derived ACA-D variants contain one or two mutations each, affecting a total of six residues. Four of these mutations, I180T, A246G, I347S, and Y354H, cluster in the virion-distal lobe of sigma3. Linkage of these mutations to AC resistance was confirmed in experiments using reovirus disassembly intermediates recoated with wt or mutant sigma3 proteins. In comparison to wt virions, ACA-D viruses displayed enhanced susceptibility to proteolysis by endocytic protease cathepsin L. Image reconstructions of cryoelectron micrographs of three ACA-D viruses that each contain a single mutation in the virion-distal lobe of sigma3 demonstrated native capsid protein organization and minimal alterations in sigma3 structure. These results suggest that mutations in sigma3 that confer resistance to inhibitors of vacuolar acidification identify a specific domain that regulates proteolytic disassembly. PMID- 16378971 TI - Live-cell characterization and analysis of a clinical isolate of bovine respiratory syncytial virus, using molecular beacons. AB - Understanding viral pathogenesis is critical for prevention of outbreaks, development of antiviral drugs, and biodefense. Here, we utilize molecular beacons to directly detect the viral genome and characterize a clinical isolate of bovine respiratory syncytial virus (bRSV) in living cells. Molecular beacons are dual-labeled, hairpin oligonucleotide probes with a reporter fluorophore at one end and a quencher at the other; they are designed to fluoresce only when hybridizing to a complementary target. By imaging the fluorescence signal of molecular beacons, the spread of bRSV was monitored for 7 days with a signal-to noise ratio of 50 to 200, and the measured time course of infection was quantified with a mathematical model for viral growth. We found that molecular beacon signal could be detected in single living cells infected with a viral titer of 2 x 10(3.6) 50% tissue culture infective doses/ml diluted 1,000 fold, demonstrating high detection sensitivity. Low background in uninfected cells and simultaneous staining of fixed cells with molecular beacons and antibodies showed high detection specificity. Furthermore, using confocal microscopy to image the viral genome in live, infected cells, we observed a connected, highly three dimensional, amorphous inclusion body structure not seen in fixed cells. Taken together, the use of molecular beacons for active virus imaging provides a powerful tool for rapid viral infection detection, the characterization of RNA viruses, and the design of new antiviral drugs. PMID- 16378972 TI - Defining the cellular target(s) of porcine reproductive and respiratory syndrome virus blocking monoclonal antibody 7G10. AB - We produced a monoclonal antibody (MAb) (7G10) that has blocking activity against porcine reproductive and respiratory syndrome virus (PRRSV). In this study, we identified the components of the 7G10 MAb-bound complex as cytoskeletal filaments: vimentin, cytokeratin 8, cytokeratin 18, actin, and hair type II basic keratin. Vimentin bound to PRRSV nucleocapsid protein and anti-vimentin antibodies showed PRRSV-blocking activity. Vimentin was expressed on the surface of MARC-145, a PRRSV-susceptible cell line. Simian vimentin rendered BHK-21 and CRFK, nonsusceptible cell lines, susceptible to PRRSV infection. These results suggest that vimentin is part of the PRRSV receptor complex and that it plays an important role in PRRSV binding with the other cytoskeletal filaments that mediate transportation of the virus in the cytosol. PMID- 16378973 TI - Kaposi's sarcoma-associated herpesvirus-encoded latency-associated nuclear antigen induces chromosomal instability through inhibition of p53 function. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is predominantly associated with three human malignancies, KS, primary effusion lymphoma, and multicentric Castleman's disease. These disorders are linked to genomic instability, known to be a crucial component of the oncogenic process. Latency-associated nuclear antigen (LANA), encoded by open reading frame 73 of the KSHV genome, is a latent protein consistently expressed in all KSHV-associated diseases. LANA is important in viral genome maintenance and is associated with cellular and viral proteins to regulate viral and cellular gene expression. LANA interacts with the tumor suppressor genes p53 and pRb, indicating that LANA may target these proteins and promote oncogenesis. In this study, we generated cell lines which stably expressed LANA to observe the effects of LANA expression on cell phenotype. LANA expression in these stable cell lines showed a dramatic increase in chromosomal instability, indicated by the presence of increased multinucleation, micronuclei, and aberrant centrosomes. In addition, these stable cell lines demonstrated an increased proliferation rate and as well as increased entry into S phase in both stable and transiently transfected LANA-expressing cells. Additionally, p53 transcription and its transactivation activity were suppressed by LANA expression in a dose-dependent manner. LANA may therefore promote chromosomal instability by suppressing the functional activities of p53, thereby facilitating KSHV-mediated pathogenesis and cancer. PMID- 16378974 TI - Human cytomegalovirus entry into epithelial and endothelial cells depends on genes UL128 to UL150 and occurs by endocytosis and low-pH fusion. AB - Human cytomegalovirus (HCMV) replication in epithelial and endothelial cells appears to be important in virus spread, disease, and persistence. It has been difficult to study infection of these cell types because HCMV laboratory strains (e.g., AD169 and Towne) have lost their ability to infect cultured epithelial and endothelial cells during extensive propagation in fibroblasts. Clinical strains of HCMV (e.g., TR and FIX) possess a cluster of genes (UL128 to UL150) that are largely mutated in laboratory strains, and recent studies have indicated that these genes facilitate replication in epithelial and endothelial cells. The mechanisms by which these genes promote infection of these two cell types are unclear. We derived an HCMV UL128-to-UL150 deletion mutant from strain TR, TRdelta4, and studied early events in HCMV infection of epithelial and endothelial cells, and the role of genes UL128 to UL150. Analysis of wild-type TR indicated that HCMV enters epithelial and endothelial cells by endocytosis followed by low-pH-dependent fusion, which is different from the pH-independent fusion with the plasma membrane observed with human fibroblasts. TRdelta4 displayed a number of defects in early infection processes. Adsorption and entry of TRdelta4 on epithelial cells were poor compared with those of TR, but these defects could be overcome with higher doses of virus and the use of polyethylene glycol (PEG) to promote fusion between virion and cellular membranes. High multiplicity and PEG treatment did not promote infection of endothelial cells by TRdelta4, yet virus particles were internalized. Together, these data indicate that genes UL128 to UL150 are required for HCMV adsorption and penetration of epithelial cells and to promote some early stage of virus replication, subsequent to virus entry, in endothelial cells. PMID- 16378975 TI - Identification of 5' and 3' cis-acting elements of the porcine reproductive and respiratory syndrome virus: acquisition of novel 5' AU-rich sequences restored replication of a 5'-proximal 7-nucleotide deletion mutant. AB - We here demonstrate the successful engineering of the RNA genome of porcine reproductive and respiratory syndrome virus (PRRSV) by using an infectious cDNA as a bacterial artificial chromosome. Runoff transcription from this cDNA by SP6 polymerase resulted in capped synthetic RNAs bearing authentic 5' and 3' ends of the viral genome that had specific infectivities of >5 x 10(5) PFU/microg of RNA. The synthetic viruses recovered from the transfected cells were genotypically and phenotypically indistinguishable from the parental virus. Using our system, a series of genomic RNAs with nucleotide deletions in their 5' ends produced viruses with decreased or no infectivity. Various pseudorevertants were isolated, and acquisition of novel 5' sequences of various sizes, composed predominantly of A and U bases, restored their infectivities, providing a novel insight into functional elements of the 5' end of the PRRSV genome. In addition, our system was further engineered to generate a panel of self-replicating, self-limiting, luciferase-expressing PRRSV viral replicons bearing various deletions. Analysis of these replicons revealed the presence and location of a 3' cis-acting element in the genome that was required for replication. Moreover, we produced enhanced green fluorescent protein-expressing infectious viruses, which indicates that the PRRSV cDNA/viral replicon/recombinant virus can be developed as a vector for the expression of a variety of heterologous genes. Thus, our PRRSV reverse genetics system not only offers a means of directly investigating the molecular mechanisms of PRRSV replication and pathogenesis but also can be used to generate new heterologous gene expression vectors and genetically defined antiviral vaccines. PMID- 16378976 TI - Characterization of Ross River virus tropism and virus-induced inflammation in a mouse model of viral arthritis and myositis. AB - Mosquito-borne alphaviruses are a significant cause of both encephalitic and arthritic disease in humans worldwide. In contrast to the encephalitic alphaviruses, the pathogenesis of alphavirus-induced arthritic disease is not well understood. Utilizing a mouse model of Ross River virus (RRV) disease, we found that the primary targets of RRV infection are bone, joint, and skeletal muscle tissues of the hind limbs in both outbred CD-1 mice and adult C57BL/6J mice. Moreover, histological analyses demonstrated that RRV infection resulted in severe inflammation of these tissues. Characterization of the inflammatory infiltrate within the skeletal muscle tissue identified inflammatory macrophages, NK cells, and CD4+ and CD8+ T lymphocytes. To determine the contribution of the adaptive immune system, the outcome of RRV-induced disease was examined in C57BL/6J RAG-1(-/-) mice, which lack functional T and B lymphocytes. RAG-1(-/-) and wild-type mice developed similar disease signs, infiltration of inflammatory macrophages and NK cells, and muscle pathology, suggesting that the adaptive immune response does not play a critical role in the development of disease. These results establish the mouse model of RRV disease as a useful system for the identification of viral and host factors that contribute to alphavirus-induced arthritis and myositis. PMID- 16378977 TI - Human immunodeficiency virus type 1 coreceptor switching: V1/V2 gain-of-fitness mutations compensate for V3 loss-of-fitness mutations. AB - Human immunodeficiency virus type 1 (HIV-1) entry into target cells is mediated by the virus envelope binding to CD4 and the conformationally altered envelope subsequently binding to one of two chemokine receptors. HIV-1 envelope glycoprotein (gp120) has five variable loops, of which three (V1/V2 and V3) influence the binding of either CCR5 or CXCR4, the two primary coreceptors for virus entry. Minimal sequence changes in V3 are sufficient for changing coreceptor use from CCR5 to CXCR4 in some HIV-1 isolates, but more commonly additional mutations in V1/V2 are observed during coreceptor switching. We have modeled coreceptor switching by introducing most possible combinations of mutations in the variable loops that distinguish a previously identified group of CCR5- and CXCR4-using viruses. We found that V3 mutations entail high risk, ranging from major loss of entry fitness to lethality. Mutations in or near V1/V2 were able to compensate for the deleterious V3 mutations and may need to precede V3 mutations to permit virus survival. V1/V2 mutations in the absence of V3 mutations often increased the capacity of virus to utilize CCR5 but were unable to confer CXCR4 use. V3 mutations were thus necessary but not sufficient for coreceptor switching, and V1/V2 mutations were necessary for virus survival. HIV 1 envelope sequence evolution from CCR5 to CXCR4 use is constrained by relatively frequent lethal mutations, deep fitness valleys, and requirements to make the right amino acid substitution in the right place at the right time. PMID- 16378978 TI - A membrane-destabilizing peptide in capsid protein L2 is required for egress of papillomavirus genomes from endosomes. AB - Papillomaviruses are internalized via clathrin-dependent endocytosis. However, the mechanism by which viral genomes pass endosomal membranes has not been elucidated. In this report we show that the minor capsid protein L2 is required for egress of viral genomes from endosomes but not for initial uptake and uncoating and that a 23-amino-acid peptide at the C terminus of L2 is necessary for this function. Pseudogenomes encapsidated by L1 and L2 lacking this peptide accumulated in vesicular compartments similar to that observed with L1-only viral particles, and these mutant pseudoviruses were noninfectious. This L2 peptide displayed strong membrane-disrupting activity, induced cytolysis of bacteria and eukaryotic cells in a pH-dependent manner, and permeabilized cells after exogenous addition. Fusions between green fluorescent protein and the L2 peptide integrated into cellular membranes like the wild type but not like C-terminal mutants of L2. Our data indicate that the L2 C terminus facilitates escape of viral genomes from the endocytic compartment and that this feature is conserved among papillomaviruses. Furthermore, the characteristic of this peptide differs from the classical virus-encoded membrane-penetrating peptides. PMID- 16378980 TI - 7a protein of severe acute respiratory syndrome coronavirus inhibits cellular protein synthesis and activates p38 mitogen-activated protein kinase. AB - It was recently shown that the 7a protein of severe acute respiratory syndrome coronavirus induces biochemical changes associated with apoptosis. In this study, the mechanism by which the 7a protein induces apoptosis was examined. The 7a protein was tested for the ability to inhibit cellular gene expression because several proapoptotic viral proteins with this function have previously been identified. 7a protein inhibited expression of luciferase from an mRNA construct that specifically measures translation, whereas inhibitors of transcription and nucleocytoplasmic transport did not. The inhibition of translation and other cellular processes of gene expression have been associated with the induction of a stress response in cells. Western blot analysis using phosphospecific antibodies indicated that 7a protein activated p38 mitogen-activated protein kinase (MAPK), but not c-Jun N-terminal protein kinase/stress-activated protein kinase. Taken together, these data indicate that the induction of apoptosis by the 7a protein may be related to its ability to inhibit cellular translation and activate p38 MAPK. PMID- 16378979 TI - UL54-null pseudorabies virus is attenuated in mice but productively infects cells in culture. AB - The pseudorabies virus (PRV) UL54 homologs are important multifunctional proteins with roles in shutoff of host protein synthesis, transactivation of virus and cellular genes, and regulation of splicing and translation. Here we describe the first genetic characterization of UL54. We constructed UL54 null mutations in a PRV bacterial artificial chromosome using sugar suicide and lambdaRed allele exchange systems. Surprisingly, UL54 is dispensable for growth in tissue culture but exhibits a small-plaque phenotype that can be complemented in trans by both the herpes simplex virus type 1 ICP27 and varicella-zoster virus open reading frame 4 proteins. Deletion of UL54 in the virus vJSdelta54 had no effect on the ability of the virus to shut off host cell protein synthesis but did affect virus gene expression. The glycoprotein gC accumulated to lower levels in cells infected with vJSdelta54 compared to those infected with wild-type virus, while gK levels were undetectable. Other late gene products, gB, gE, and Us9, accumulated to higher levels than those seen in cells infected with wild-type virus in a multiplicity-dependent manner. DNA replication is also reduced in cells infected with vJSdelta54. UL54 appears to regulate UL53 and UL52 at the transcriptional level as their respective RNAs are decreased in cells infected with vJSdelta54. Interestingly, vJSdelta54 is highly attenuated in a mouse model of PRV infection. Animals infected with vJSdelta54 survive twice as long as animals infected with wild-type virus, and this results in delayed accumulation of virus-specific antigens in skin, dorsal root ganglia, and spinal cord tissues. PMID- 16378981 TI - Involvement of template-activating factor I/SET in transcription of adenovirus early genes as a positive-acting factor. AB - The adenovirus genome complexed with viral core protein VII (adenovirus DNA protein VII complex) at least is the bona fide template for transcription of adenovirus early genes. It is believed that the highly basic protein VII, like cellular histones, is a negative regulator for genome functions. Analyses with in vitro replication and transcription systems using the adenovirus DNA-protein VII complex have revealed that remodeling of the complex is crucial for efficient DNA replication and transcription. We identified host acidic proteins, template activating factor I (TAF-I), TAF-II, and TAF-III as stimulatory factors for replication from the adenovirus DNA-protein VII complex. Recently, it was reported that the adenovirus DNA interacts with TAF-I and pp32, another host acidic protein (Y. Xue, J. S. Johnson, D. A. Ornelles, J. Lieberman, and D. A. Engel, J. Virol. 79:2474-2483, 2005). We found that TAF-I interacts and colocalizes with protein VII in adenovirus-infected cells during the early phases of infection, but pp32 does not. Although pp32 had the potential ability to interact with protein VII, pp32 did not remodel the adenovirus DNA-protein VII complex in vitro. Small interfering RNA-mediated knockdown of TAF-I expression leads to the delay of the transcription timing of early genes. These results provide evidence that TAF-I plays an important role in the early stages of the adenovirus infection cycle. PMID- 16378982 TI - Naive and memory cell turnover as drivers of CCR5-to-CXCR4 tropism switch in human immunodeficiency virus type 1: implications for therapy. AB - Early human immunodeficiency virus infection is characterized by the predominance of CCR5-tropic (R5) virus. However, in many individuals CXCR4-tropic (X4) virus appears in late infection. The reasons for this phenotypic switch are unclear. The patterns of chemokine receptor expression suggest that X4 and R5 viruses have a preferential tropism for naive and memory T cells, respectively. Since memory cells divide approximately 10 times as often as naive cells in uninfected individuals, a tropism for memory cells in early infection may provide an advantage. However, with disease progression both naive and memory cell division frequencies increase, and at low CD4 counts, the naive cell division frequency approaches that of memory cells. This may provide a basis for the phenotypic switch from R5 to X4 virus observed in late infection. We show that a model of infection using observed values for cell turnover supports this mechanism. The phenotypic switch from R5 to X4 virus occurs at low CD4 counts and is accompanied by a rapid rise in viral load and drop in CD4 count. Thus, low CD4 counts are both a cause and an effect of X4 virus dominance. We also investigate the effects of different antiviral strategies. Surprisingly, these results suggest that both conventional antiretroviral regimens and CCR5 receptor-blocking drugs will promote R5 virus over X4 virus. PMID- 16378983 TI - Impact of capsid conformation and Rep-capsid interactions on adeno-associated virus type 2 genome packaging. AB - Single-stranded genomes of adeno-associated virus (AAV) are packaged into preformed capsids. It has been proposed that packaging is initiated by interaction of genome-bound Rep proteins to the capsid, thereby targeting the genome to the portal of encapsidation. Here we describe a panel of mutants with amino acid exchanges in the pores at the fivefold axes of symmetry on AAV2 capsids with reduced packaging and reduced Rep-capsid interaction. Mutation of two threonines at the rim of the fivefold pore nearly completely abolished Rep capsid interaction and packaging. This suggests a Rep-binding site at the highly conserved amino acids at or close to the pores formed by the capsid protein pentamers. A different mutant (P. Wu, W. Xiao, T. Conlon, J. Hughes, M. Agbandje McKenna, T. Ferkol, T. Flotte, and N. Muzyczka, J. Virol. 74:8635-8647, 2000) with an amino acid exchange at the interface of capsid protein pentamers led to a complete block of DNA encapsidation. Analysis of the capsid conformation of this mutant revealed that the pores at the fivefold axes were occupied by VP1/VP2 N termini, thereby preventing DNA introduction into the capsid. Nevertheless, the corresponding capsids had more Rep proteins bound than wild-type AAV, showing that correct Rep interaction with the capsid depends on a defined capsid conformation. Both mutant types together support the conclusion that the pores at the fivefold symmetry axes are involved in genome packaging and that capsid conformation-dependent Rep-capsid interactions play an essential role in the packaging process. PMID- 16378984 TI - Mutations on the external surfaces of adeno-associated virus type 2 capsids that affect transduction and neutralization. AB - Mutations were made at 64 positions on the external surface of the adeno associated virus type 2 (AAV-2) capsid in regions expected to bind antibodies. The 127 mutations included 57 single alanine substitutions, 41 single nonalanine substitutions, 27 multiple mutations, and 2 insertions. Mutants were assayed for capsid synthesis, heparin binding, in vitro transduction, and binding and neutralization by murine monoclonal and human polyclonal antibodies. All mutants made capsid proteins within a level about 20-fold of that made by the wild type. All but seven mutants bound heparin as well as the wild type. Forty-two mutants transduced human cells at least as well as the wild type, and 10 mutants increased transducing activity up to ninefold more than the wild type. Eighteen adjacent alanine substitutions diminished transduction from 10- to 100,000-fold but had no effect on heparin binding and define an area (dead zone) required for transduction that is distinct from the previously characterized heparin receptor binding site. Mutations that reduced binding and neutralization by a murine monoclonal antibody (A20) were localized, while mutations that reduced neutralization by individual human sera or by pooled human, intravenous immunoglobulin G (IVIG) were dispersed over a larger area. Mutations that reduced binding by A20 also reduced neutralization. However, a mutation that reduced the binding of IVIG by 90% did not reduce neutralization, and mutations that reduced neutralization by IVIG did not reduce its binding. Combinations of mutations did not significantly increase transduction or resistance to neutralization by IVIG. These mutations define areas on the surface of the AAV-2 capsid that are important determinants of transduction and antibody neutralization. PMID- 16378985 TI - Neutralization escape variants of human immunodeficiency virus type 1 are transmitted from mother to infant. AB - Maternal passive immunity typically plays a critical role in protecting infants from new infections; however, the specific contribution of neutralizing antibodies in limiting mother-to-child transmission of human immunodeficiency virus type 1 is unclear. By examining cloned envelope variants from 12 transmission pairs, we found that vertically transmitted variants were more resistant to neutralization by maternal plasma than were maternal viral variants near the time of transmission. The vertically transmitted envelope variants were poorly neutralized by monoclonal antibodies b12 [corrected] 2G12, 2F5, and 4E10 individually or in combination. Despite the fact that the infant viruses were among the most neutralization resistant in the mother, they had relatively few glycosylation sites. Moreover, the transmitted variants elicited de novo neutralizing antibodies in the infants, indicating that they were not inherently difficult to neutralize. The neutralization resistance of vertically transmitted viruses is in contrast to the relative neutralization sensitivity of viruses sexually transmitted within discordant couples, suggesting that the antigenic properties of viruses that are favored for transmission may differ depending upon mode of transmission. PMID- 16378986 TI - Epstein-Barr virus (EBV) genome and expression in breast cancer tissue: effect of EBV infection of breast cancer cells on resistance to paclitaxel (Taxol). AB - The Epstein-Barr virus (EBV) has been detected in subsets of breast cancers. In order to elaborate on these observations, we quantified by real-time PCR (Q-PCR) the EBV genome in biopsy specimens of breast cancer tissue as well as in tumor cells isolated by microdissection. Our findings show that EBV genomes can be detected by Q-PCR in about half of tumor specimens, usually in low copy numbers. However, we also found that the viral load is highly variable from tumor to tumor. Moreover, EBV genomes are heterogeneously distributed in morphologically identical tumor cells, with some clusters of isolated tumor cells containing relatively high genome numbers while other tumor cells isolated from the same specimen may be negative for EBV DNA. Using reverse transcription-PCR, we detected EBV gene transcripts: EBNA-1 in almost all of the EBV-positive tumors and RNA of the EBV oncoprotein LMP-1 in a smaller subset of the tissues analyzed. Moreover, BARF-1 RNA was detected in half of the cases studied. Furthermore, we observed that in vitro EBV infection of breast carcinoma cells confers resistance to paclitaxel (taxol) and provokes overexpression of a multidrug resistance gene (MDR1). Consequently, even if a small number of breast cancer cells are EBV infected, the impact of EBV infection on the efficiency of anticancer treatment might be of importance. PMID- 16378987 TI - R5 variants of human immunodeficiency virus type 1 preferentially infect CD62L- CD4+ T cells and are potentially resistant to nucleoside reverse transcriptase inhibitors. AB - The persistence of human immunodeficiency virus type 1 (HIV-1) in memory CD4+ T cells is a major obstacle to the eradication of the virus with current antiretroviral therapy. Here, we investigated the effect of the activation status of CD4+ T cells on the predominance of R5 and X4 HIV-1 variants in different subsets of CD4+ T cells in ex vivo-infected human lymphoid tissues and peripheral blood mononuclear cells (PBMCs). In these cell systems, we examined the sensitivity of HIV replication to reverse transcriptase inhibitors. We demonstrate that R5 HIV-1 variants preferentially produced productive infection in HLA-DR- CD62L- CD4+ T cells. These cells were mostly in the G1b phase of the cell cycle, divided slowly, and expressed high levels of CCR5. In contrast, X4 HIV-1 variants preferentially produced productive infection in activated HLA-DR+ CD62L+ CD4+ T cells, which expressed high levels of CXCR4. The abilities of the nucleoside reverse transcriptase inhibitors (NRTI) zidovudine and lamivudine to stop HIV-1 replication were 20 times greater in activated T cells than in slowly dividing HLA-DR- CD62L- CD4+ T cells. This result, demonstrated both in a highly physiologically relevant ex vivo lymphoid tissue model and in PBMCs, correlated with higher levels of thymidine kinase mRNA in activated than in slowly dividing HLA-DR- CD62L- CD4+ T cells. The non-NRTI nevirapine was equally efficient in both cell subsets. The lymphoid tissue and PBMC-derived cell systems represent well-defined models which could be used as new tools for the study of the mechanism of resistance to HIV-1 inhibitors in HLA-DR- CD62L- CD4+ T cells. PMID- 16378989 TI - Uracil DNA glycosylase is dispensable for human immunodeficiency virus type 1 replication and does not contribute to the antiviral effects of the cytidine deaminase Apobec3G. AB - It is well established that many host factors are involved in the replication of human immunodeficiency virus (HIV) type 1. One host protein, uracil DNA glycosylase 2 (UNG2), binds to multiple viral proteins and is packaged into HIV type 1 virions. UNG initiates the removal of uracils from DNA, and this has been proposed to be important both for reverse transcription and as a mediator to the antiviral effect of virion-incorporated Apobec3G, a cytidine deaminase that generates numerous uracils in the viral DNA during virus replication. We used a natural human UNG-/- cell line as well as cells that express a potent catalytic active-site inhibitor of UNG to assess the effects of removing UNG activity on HIV infectivity. In both cases, we find UNG2 activity and protein to be completely dispensable for virus replication. Moreover, we find that virion associated UNG2 does not affect the loss of infectivity caused by Apobec3G. PMID- 16378988 TI - Hepatitis C virus induces toll-like receptor 4 expression, leading to enhanced production of beta interferon and interleukin-6. AB - Hepatitis C virus (HCV) induces inflammatory signals, leading to hepatitis, hepatocellular carcinomas, and lymphomas. The mechanism of HCV involvement in the host's innate immune responses has not been well characterized. In this study, we analyzed expression and regulation of the entire panel of toll-like receptors (TLRs) in human B cells following HCV infection in vitro. Among all of the TLRs (TLRs 1 to 10) examined, only TLR4 showed an altered expression (a three- to sevenfold up-regulation) after HCV infection. Peripheral blood mononuclear cells from HCV-infected individuals also showed a higher expression level of TLR4 compared with those of healthy individuals. HCV infection significantly increased beta interferon (IFN-beta) and interleukin-6 (IL-6) secretion from B cells, particularly after lipopolysaccharide stimulation. The increased IFN-beta and IL 6 production was mediated by TLR4 induction, since the introduction of the small interfering RNA against TLR4 specifically inhibited the HCV-induced cytokine production. Among all of the viral proteins, only NS5A caused TLR4 induction in hepatocytes and B cells. NS5A specifically activated the promoter of the TLR4 gene in both hepatocytes and B cells. In conclusion, HCV infection directly induces TLR4 expression and thereby activates B cells, which may contribute to the host's innate immune responses. PMID- 16378990 TI - Three immunoproteasome-associated subunits cooperatively generate a cytotoxic T lymphocyte epitope of Epstein-Barr virus LMP2A by overcoming specific structures resistant to epitope liberation. AB - The precise roles of gamma interferon-inducible immunoproteasome-associated molecules in generation of cytotoxic T-lymphocyte (CTL) epitopes have yet to be fully elucidated. We describe here a unique epitope derived from the Epstein-Barr virus (EBV) latent membrane protein 2A (LMP2A) presented by HLA-A*2402 molecules. Generation of the epitope, designated LMP2A(222-230), from the full-length protein requires the immunoproteasome subunit low-molecular-weight protein 7 (ip LMP7) and the proteasome activator 28-alpha subunit and is accelerated by ip LMP2, as revealed by gene expression experiments using an LMP2A(222-230)-specific CTL clone as a responder in enzyme-linked immunospot assays. The unequivocal involvement of all three components was confirmed by RNA interference gene silencing. Interestingly, the LMP2A(222-230) epitope could be efficiently generated from incomplete EBV-LMP2A fragments that were produced by puromycin treatment or gene-engineered shortened EBV-LMP2A lacking some of its hydrophobic domains. In addition, epitope generation was increased by a single amino acid substitution from leucine to alanine immediately flanking the C terminus, this being predicted by a web-accessible program to increase the cleavage strength. Taken together, the data indicate that the generation of LMP2A(222-230) is influenced not only by extrinsic factors such as immunoproteasomes but also by intrinsic factors such as the length of the EBV-LMP2A protein and proteasomal cleavage strength at specific positions in the source antigen. PMID- 16378991 TI - Potent neutralization of Hendra and Nipah viruses by human monoclonal antibodies. AB - Hendra virus (HeV) and Nipah virus (NiV) are closely related emerging viruses comprising the Henipavirus genus of the Paramyxovirinae. Each has a broad species tropism and can cause disease with high mortality in both animal and human hosts. These viruses infect cells by a pH-independent membrane fusion event mediated by their attachment (G) and fusion (F) envelope glycoproteins (Envs). Seven Fabs, m101 to -7, were selected for their significant binding to a soluble form of Hendra G (sG) which was used as the antigen for panning of a large naive human antibody library. The selected Fabs inhibited, to various degrees, cell fusion mediated by the HeV or NiV Envs and virus infection. The conversion of the most potent neutralizer of infectious HeV, Fab m101, to immunoglobulin G1 (IgG1) significantly increased its cell fusion inhibitory activity: the 50% inhibitory concentration was decreased more than 10-fold to approximately 1 microg/ml. The IgG1 m101 was also exceptionally potent in neutralizing infectious HeV; complete (100%) neutralization was achieved with 12.5 microg/ml, and 98% neutralization required only 1.6 microg/ml. The inhibition of fusion and infection correlated with binding of the Fabs to full-length G as measured by immunoprecipitation and less with binding to sG as measured by enzyme-linked immunosorbent assay and Biacore. m101 and m102 competed with the ephrin-B2, which we recently identified as a functional receptor for both HeV and NiV, indicating a possible mechanism of neutralization by these antibodies. The m101, m102, and m103 antibodies competed with each other, suggesting that they bind to overlapping epitopes which are distinct from the epitopes of m106 and m107. In an initial attempt to localize the epitopes of m101 and m102, we measured their binding to a panel of 11 G alanine-scanning mutants and identified two mutants, P185A and Q191 K192A, which significantly decreased binding to m101 and one, G183, which decreased binding of m102 to G. These results suggest that m101 to -7 are specific for HeV or NiV or both and exhibit various neutralizing activities; they are the first human monoclonal antibodies identified against these viruses and could be used for treatment, prophylaxis, and diagnosis and as research reagents and could aid in the development of vaccines. PMID- 16378992 TI - The amino-terminal domain of bovine viral diarrhea virus Npro protein is necessary for alpha/beta interferon antagonism. AB - The alpha/beta interferon (IFN-alpha/beta) system is the first line of defense against viral infection and a critical link between the innate and adaptive immune responses. IFN-alpha/beta secretion is the hallmark of cellular responses to acute RNA virus infections. As part of their survival strategy, many viruses have evolved mechanisms to counteract the host IFN-alpha/beta response. Bovine viral diarrhea virus (BVDV) (genus Pestivirus) was reported to trigger interferon production in infected cultured cells under certain circumstances or to suppress it under others. Our studies with various cultured fibroblasts and epithelial bovine cells indicated that cytopathic (cp) BVDV induces IFN-alpha/beta very inefficiently. Using a set of engineered cp BVDVs expressing mutant Npro and appropriate controls, we found that the IFN-alpha/beta response to infection was dependent on Npro expression and independent of viral replication efficiency. In order to investigate whether the protease activity of Npro is required for IFN alpha/beta antagonism, we engineered Npro mutants lacking protease activity by replacement of amino acid E22, H49, or C69. We found that E22 and H49 substitutions abolished the ability of Npro to suppress IFN, whereas C69 had no effect, suggesting that the structural integrity of the N terminus of Npro was more important than its catalytic activity for IFN-alpha/beta suppression. A catalytically active mutant with a change at a conserved Npro region near the N terminus (L8P) in both BVDV biotypes did not antagonize IFN-alpha/beta production, confirming its involvement in this process. Taken together, these results not only provide direct evidence for the role of Npro in blocking IFN alpha/beta induction, but also implicate the amino-terminal domain of the protein in this function. PMID- 16378994 TI - Human cytomegalovirus immediate-early 2 protein IE86 blocks virus-induced chemokine expression. AB - The effect of human cytomegalovirus (HCMV) gene expression on cytokine (beta interferon) and chemokine (RANTES, MIG, MCP-2, MIP-1alpha, and interleukin-8) expression was examined. We demonstrate that HCMV gene expression is required to suppress the transcriptional induction of these cytokines and that the HCMV immediate-early 2 gene product IE86 can effectively block the expression of cytokines and proinflammatory chemokines during HCMV and Sendai virus infection. Additionally, we present data on viral mutants and ectopic protein expression which demonstrate that pp65, another identified HCMV cytokine antagonist, is not involved in regulating these proinflammatory cytokines. This is the first report to demonstrate that IE86 can act to suppress virus-induced proinflammatory cytokine transcript expression, extending the antiviral properties of this multifunctional viral protein. PMID- 16378993 TI - Immune response to individual maedi-visna virus gag antigens. AB - The lesions caused by maedi-visna virus (MVV) are known to be immune mediated with a presumed contribution by the response to viral antigens. However, very little is known about the T-cell response to individual viral proteins. We have therefore expressed the three individual gag antigens of MVV strain EV1 (p16, p25, and p14) in a bacterial expression system and used the purified recombinant proteins to analyze the antibody and CD4+ T-cell response to MVV. Plasma samples were taken from sheep after 1 year of infection with MVV. The titers for antibodies in these samples were determined by indirect enzyme-linked immunosorbent assays and were as follows: anti-p25 antibody, 1:400 to >1:3,200; anti-p16 antibody, 1:400 to 1:3,200; and anti-p14 antibody, 1:<100 to 1:3,200. When the induction of antibodies was followed over time postinfection (p.i.), samples positive for anti-p25 were seen by day 24 p.i., followed by anti-p16 by day 45 p.i., and lastly anti-p14 by day 100 p.i. T-cell proliferative responses to all three gag antigens were detected in persistently infected sheep peripheral blood lymphocytes. The antigens were therefore used to raise T-cell lines from persistently infected sheep. These T-cell lines were shown to be specific for the recombinant gag antigens and for viral antigen expressed on infected macrophages. The proliferative response was restricted to major histocompatibility complex class II HLA-DR and so was due to CD4+ T lymphocytes. All three gag antigens may therefore play a role in immune-mediated lesion formation in MVV disease by presentation on infected macrophages in lesions. PMID- 16378995 TI - Functional analysis of the triplex proteins (VP19C and VP23) of herpes simplex virus type 1. AB - The triplex of herpesvirus capsids is a unique structural element. In herpes simplex virus type 1 (HSV-1), one molecule of VP19C and two of VP23 form a three pronged structure that acts to stabilize the capsid shell through interactions with adjacent VP5 molecules. The interaction between VP19C and VP23 was inferred by yeast cryoelectron microscopy studies and subsequently confirmed by the two hybrid assay. In order to define the functional domains of VP19C and VP23, a Tn7 based transposon was used to randomly insert 15 bp into the coding regions of these two proteins. The mutants were initially screened for interaction in the yeast two-hybrid assay to identify the domains important for triplex formation. Using genetic complementation assays in HSV-1-infected cells, the domains of each protein required for virus replication were similarly uncovered. The same mutations that abolish interaction between these two proteins in the yeast two hybrid assay similarly failed to complement the growth of the VP23- and VP19C null mutant viruses in the genetic complementation assay. Some of these mutants were transferred into recombinant baculoviruses to analyze the effect of the mutations on herpesvirus capsid assembly in insect cells. The mutations that abolished the interaction in the yeast two-hybrid assay also abolished capsid assembly in insect cells. The outcome of these experiments showed that insertions in at least four regions and especially the amino terminus of VP23 abolished function, whereas the amino terminus of VP19C can tolerate transposon insertions. A novel finding of these studies was the ability to assemble herpesvirus capsids in insect cells using VP5 and VP19C that contained a histidine handle at their amino terminus. PMID- 16378996 TI - Monoclonal antibodies targeting the HR2 domain and the region immediately upstream of the HR2 of the S protein neutralize in vitro infection of severe acute respiratory syndrome coronavirus. AB - We have previously shown that an Escherichia coli-expressed, denatured spike (S) protein fragment of the severe acute respiratory coronavirus, containing residues 1029 to 1192 which include the heptad repeat 2 (HR2) domain, was able to induce neutralizing polyclonal antibodies (C. T. Keng, A. Zhang, S. Shen, K. M. Lip, B. C. Fielding, T. H. Tan, C. F. Chou, C. B. Loh, S. Wang, J. Fu, X. Yang, S. G. Lim, W. Hong, and Y. J. Tan, J. Virol. 79:3289-3296, 2005). In this study, monoclonal antibodies (MAbs) were raised against this fragment to identify the linear neutralizing epitopes in the functional domain and to investigate the mechanisms involved in neutralization. Eighteen hybridomas secreting the S protein-specific MAbs were obtained. Binding sites of these MAbs were mapped to four linear epitopes. Two of them were located within the HR2 region and two immediately upstream of the HR2 domain. MAbs targeting these epitopes showed in vitro neutralizing activities and were able to inhibit cell-cell membrane fusion. These results provide evidence of novel neutralizing epitopes that are located in the HR2 domain and the spacer region immediately upstream of the HR2 of the S protein. PMID- 16378997 TI - Human cytomegalovirus protein pp71 disrupts major histocompatibility complex class I cell surface expression. AB - The human cytomegalovirus tegument protein pp71 is the product of the UL82 gene. Roles for pp71 in stimulating gene transcription, increasing infectivity of viral DNA, and the degradation of retinoblastoma family proteins have been described. Here we report a novel function for pp71 in limiting accumulation of cell surface major histocompatibility complex (MHC) class I complexes. MHC molecules were analyzed in glioblastoma cells exposed to a replication-defective adenovirus expressing UL82 (Adpp71) or after transient transfection of the UL82 gene. Accumulation of cell surface MHC class I levels diminished in a specific and dose dependent manner after exposure to Adpp71 but not after exposure to an adenovirus expressing beta-galactosidase (Adbeta gal). UL82 expression did not interfere with accumulation of either MHC class I heavy-chain transcript or protein, nor did UL82 expression correlate with markers of apoptosis. Rather, UL82 expression correlated with an increased proportion of MHC class I molecules exhibiting sensitivity to endoglycosidase H treatment. Finally, we show that, in cells infected with recombinant virus strain missing all of the unique short region MHC class I evasion genes, disruption of UL82 expression by short, interfering RNAs led to increased accumulation of cell surface MHC class I complexes. These findings support a novel role for HCMV pp71 in disruption of the MHC class I antigen presentation pathway. PMID- 16378998 TI - Adenovirus E1B 55-kilodalton protein is required for both regulation of mRNA export and efficient entry into the late phase of infection in normal human fibroblasts. AB - The human adenovirus type 5 (Ad5) E1B 55-kDa protein is required for selective nuclear export of viral late mRNAs from the nucleus and concomitant inhibition of export of cellular mRNAs in HeLa cells and some other human cell lines, but its contributions(s) to replication in normal human cells is not well understood. We have therefore examined the phenotypes exhibited by viruses carrying mutations in the E1B 55-kDa protein coding sequence in normal human fibroblast (HFFs). Ad5 replicated significantly more slowly in HFFs than it does in tumor cells, a difference that is the result of delayed entry into the late phase of infection. The A143 mutation, which specifically impaired export of viral late mRNAs from the nucleus in infected HeLa cells (R. A. Gonzalez and S. J. Flint, J. Virol. 76:4507-4519, 2002), induced a more severe defect in viral mRNA export in HFFs. This observation indicates that the E1B 55-kDa protein regulates mRNA export during the late phase of infection of normal human cells. Other mutants exhibited phenotypes not observed in HeLa cells. In HFFs infected by the null mutant Hr6, synthesis of viral late mRNAs and proteins was severely impaired. Such defects in late gene expression were the result of inefficient progression into the late phase of infection, for viral DNA synthesis was 10-fold less efficient in Hr6 infected HFFs than in cells infected by Ad5. Similar, but less severe, defects in viral DNA synthesis were induced by the insertion mutation H224, which has been reported to inhibit binding of the E1B 55-kDa protein to p53 (C. C. Kao, P. R. Yew, and A. J. Berk, Virology 179:806-814, 1990). PMID- 16378999 TI - Natural history of human respiratory syncytial virus inferred from phylogenetic analysis of the attachment (G) glycoprotein with a 60-nucleotide duplication. AB - A total of 47 clinical samples were identified during an active surveillance program of respiratory infections in Buenos Aires (BA) (1999 to 2004) that contained sequences of human respiratory syncytial virus (HRSV) with a 60 nucleotide duplication in the attachment (G) protein gene. This duplication was analogous to that previously described for other three viruses also isolated in Buenos Aires in 1999 (A. Trento et al., J. Gen. Virol. 84:3115-3120, 2003). Phylogenetic analysis indicated that BA sequences with that duplication shared a common ancestor (dated about 1998) with other HRSV G sequences reported worldwide after 1999. The duplicated nucleotide sequence was an exact copy of the preceding 60 nucleotides in early viruses, but both copies of the duplicated segment accumulated nucleotide substitutions in more recent viruses at a rate apparently higher than in other regions of the G protein gene. The evolution of the viruses with the duplicated G segment apparently followed the overall evolutionary pattern previously described for HRSV, and this genotype has replaced other prevailing antigenic group B genotypes in Buenos Aires and other places. Thus, the duplicated segment represents a natural tag that can be used to track the dissemination and evolution of HRSV in an unprecedented setting. We have taken advantage of this situation to reexamine the molecular epidemiology of HRSV and to explore the natural history of this important human pathogen. PMID- 16379000 TI - Host response to the attenuated poxvirus vector NYVAC: upregulation of apoptotic genes and NF-kappaB-responsive genes in infected HeLa cells. AB - NYVAC has been engineered as a safe, attenuated vaccinia virus (VV) vector for use in vaccination against a broad spectrum of pathogens and tumors. Due to the interest in NYVAC-based vectors as vaccines and current phase I/II clinical trials with this vector, there is a need to analyze the human host response to NYVAC infection. Using high-density cDNA microarrays, we found 368 differentially regulated genes after NYVAC infection of HeLa cells. Clustering of the regulated genes identified six discrete gene clusters with altered expression patterns. Clusters 1 to 3 represented 47.5% of the regulated genes, with three patterns of gene activation kinetics, whereas clusters 4 to 6 showed distinct repression kinetics. Quantitative real-time reverse transcription-PCR analysis of selected genes validated the array data. Upregulated transcripts correlated with genes implicated in immune responses, including those encoding interleukin-1 receptor 2 (IL-1R2), IL-6, ISG-15, CD-80, and TNFSF7. NYVAC upregulated several intermediates of apoptotic cascades, including caspase-9, correlating with its ability to induce apoptosis. NYVAC infection also stimulated the expression of NF kappaB1 and NF-kappaB2 as well as that of NF-kappaB target genes. Expression of the VV host range K1L gene during NYVAC infection prevented NF-kappaB activation, but not the induction of apoptosis. This study is the first overall analysis of the transcriptional response of human cells to NYVAC infection and provides a framework for future functional studies to evaluate this vector and its derivatives as human vaccines. PMID- 16379001 TI - Consistent patterns of change during the divergence of human immunodeficiency virus type 1 envelope from that of the inoculated virus in simian/human immunodeficiency virus-infected macaques. AB - We have analyzed changes to proviral Env gp120 sequences and the development of neutralizing antibodies (NAbs) during 1 year of simian/human immunodeficiency virus SHIV-89.6P infection in 11 Macaca nemestrina macaques. Seven macaques had significant env divergence from that of the inoculum, and macaques with greater divergence had higher titers of homologous NAbs. Substitutions in sequons encoding potential N-linked glycosylation sites (PNGs) were among the first to be established, although overall the total number of sequons did not increase significantly. The majority (19 of 23) of PNGs present in the inoculum were conserved in the sequences from all macaques. Statistically significant variations in PNGs occurred in multiple macaques within constrained regions we term "hot spots," resulting in the selection of sequences more similar to the B consensus. These included additions on V1, the N-terminal side of V4, and the outer region of C2. Complex mutational patterns resulted in convergent PNG shifts in V2 and V5. Charge changes in Env V1V2, resulting in a net acidic charge, and a proline addition in V5 occurred in several macaques. Molecular modeling of the 89.6P sequence showed that the conserved glycans lie on the silent face of Env and that many are proximal to disulfide bonds, while PNG additions and shifts are proximal to the CD4 binding site. Nonsynonymous-to-synonymous substitution ratios suggest that these changes result from selective pressure. This longitudinal and cross-sectional study of mutations in human immunodeficiency virus (HIV) env in the SHIV background provides evidence that there are more constraints on the configuration of the glycan shield than were previously appreciated. PMID- 16379002 TI - Low pH-dependent endosomal processing of the incoming parvovirus minute virus of mice virion leads to externalization of the VP1 N-terminal sequence (N-VP1), N VP2 cleavage, and uncoating of the full-length genome. AB - Minute virus of mice (MVM) enters the host cell via receptor-mediated endocytosis. Although endosomal processing is required, its role remains uncertain. In particular, the effect of low endosomal pH on capsid configuration and nuclear delivery of the viral genome is unclear. We have followed the progression and structural transitions of DNA full-virus capsids (FC) and empty capsids (EC) containing the VP1 and VP2 structural proteins and of VP2-only virus like particles (VLP) during the endosomal trafficking. Three capsid rearrangements were detected in FC: externalization of the VP1 N-terminal sequence (N-VP1), cleavage of the exposed VP2 N-terminal sequence (N-VP2), and uncoating of the full-length genome. All three capsid modifications occurred simultaneously, starting as early as 30 min after internalization, and all of them were blocked by raising the endosomal pH. In particles lacking viral single stranded DNA (EC and VLP), the N-VP2 was not exposed and thus it was not cleaved. However, the EC did externalize N-VP1 with kinetics similar to those of FC. The bulk of all the incoming particles (FC, EC, and VLP) accumulated in lysosomes without signs of lysosomal membrane destabilization. Inside lysosomes, capsid degradation was not detected, although the uncoated DNA of FC was slowly degraded. Interestingly, at any time postinfection, the amount of structural proteins of the incoming virions accumulating in the nuclear fraction was negligible. These results indicate that during the early endosomal trafficking, the MVM particles are structurally modified by low-pH-dependent mechanisms. Regardless of the structural transitions and protein composition, the majority of the entering viral particles and genomes end in lysosomes, limiting the efficiency of MVM nuclear translocation. PMID- 16379003 TI - Varicella-zoster virus ORF63 inhibits apoptosis of primary human neurons. AB - Virus-encoded modulation of apoptosis may serve as a mechanism to enhance cell survival and virus persistence. The impact of productive varicella-zoster virus (VZV) infection on apoptosis appears to be cell type specific, as infected human sensory neurons are resistant to apoptosis, yet human fibroblasts readily become apoptotic. We sought to identify the viral gene product(s) responsible for this antiapoptotic phenotype in primary human sensory neurons. Treatment with phosphonoacetic acid to inhibit viral DNA replication and late-phase gene expression did not alter the antiapoptotic phenotype, implicating immediate-early (IE) or early genes or a virion component. Compared to the parental VZV strain (rOKA), a recombinant virus unable to express one copy of the diploid IE gene ORF63 (rOka deltaORF63) demonstrated a significant induction of apoptosis in infected neurons, as determined by three methods: annexin V staining, deoxynucleotidyltransferase-mediated dUTP-biotin nick end label staining, and transmission electron microscopy. Furthermore, neurons transfected with a plasmid expressing ORF63 resisted apoptosis induced by nerve growth factor withdrawal. These results show that ORF63 can suppress apoptosis of neurons and provide the first identification of a VZV gene encoding an antiapoptotic function. As ORF63 is expressed in neurons during both productive and latent infection, it may play a significant role in viral pathogenesis by promoting neuron survival during primary and reactivated infections. PMID- 16379004 TI - Interferon regulatory factor 3 is necessary for induction of antiviral genes during human cytomegalovirus infection. AB - Viral infection activates interferon regulatory factor 3 (IRF3), a cofactor for the induction of interferon-stimulated genes (ISGs). The role of IRF3 in the activation of ISGs by human cytomegalovirus (HCMV) is controversial despite the fact that HCMV has consistently been shown to induce ISGs during infection of fibroblasts. To address the function of IRF3 in HCMV-mediated ISG induction, we monitored ISG expression and global gene expression in HCMV-infected cells in which IRF3 function had been depleted by small interfering RNA or blocked by dominant negative IRF3. A specific reduction of ISG induction was observed, whereas other transcripts were unaffected. We therefore conclude that IRF3 specifically regulates ISG induction during the initial phase of HCMV infection. PMID- 16379005 TI - Rescue of recombinant Marburg virus from cDNA is dependent on nucleocapsid protein VP30. AB - Here we report recovery of infectious Marburg virus (MARV) from a full-length cDNA clone. Compared to the wild-type virus, recombinant MARV showed no difference in terms of morphology of virus particles, intracellular distribution in infected cells, and growth kinetics. The nucleocapsid protein VP30 of MARV and Ebola virus (EBOV) contains a Zn-binding motif which is important for the function of VP30 as a transcriptional activator in EBOV, whereas its role for MARV is unclear. It has been reported previously that MARV VP30 is able to support transcription in an EBOV-specific minigenome system. When the Zn-binding motif was destroyed, MARV VP30 was shown to be inactive in the EBOV system. While it was not possible to rescue recombinant MARV when the VP30 plasmid was omitted from transfection, MARV VP30 with a destroyed Zn-binding motif and EBOV VP30 were able to mediate virus recovery. In contrast, rescue of recombinant EBOV was not supported by EBOV VP30 containing a mutated Zn-binding domain. PMID- 16379006 TI - Mefloquine, an antimalaria drug with antiprion activity in vitro, lacks activity in vivo. AB - In view of the effectiveness of antimalaria drugs inhibiting abnormal protease resistant prion protein (PrP-res) formation in scrapie agent-infected cells, we tested other antimalarial compounds for similar activity. Mefloquine (MF), a quinoline antimalaria drug, was the most active compound tested against RML and 22L mouse scrapie agent-infected cells, with 50% inhibitory concentrations of approximately 0.5 and approximately 1.2 microM, respectively. However, MF administered to mice did not delay the onset of intraperitoneally inoculated scrapie agent, the result previously observed with quinacrine. While most anti scrapie agent compounds inhibit PrP-res formation in vitro, many PrP-res inhibitors have no activity in vivo. This underscores the importance of testing promising candidates in vivo. PMID- 16379007 TI - Longitudinal analysis of human immunodeficiency virus type 1 nef/long terminal repeat sequences in a cohort of long-term survivors infected from a single source. AB - We studied the evolution of human immunodeficiency virus type 1 (HIV-1) in a cohort of long-term survivors infected with an attenuated strain of HIV-1 acquired from a single source. Although the cohort members experienced differing clinical courses, we demonstrate similar evolution of HIV-1 nef/long-terminal repeat (LTR) sequences, characterized by progressive sequence deletions tending toward a minimal nef/LTR structure that retains only sequence elements required for viral replication. The in vivo pathogenicity of attenuated HIV-1 is therefore dictated by viral and/or host factors other than those that impose a unidirectional selection pressure on the nef/LTR region of the HIV-1 genome. PMID- 16379008 TI - ATP hydrolysis and AMP kinase activities of nonstructural protein 2C of human parechovirus 1. AB - The highly conserved picornavirus 2C proteins, thought to be involved in genome replication, contain three motifs found in NTPases/helicases of superfamily III. We report that human parechovirus 1 2C displays Mg2+-dependent ATP diphosphohydrolase activity in vitro, whereas other nucleoside triphosphates are not substrates for the hydrolysis. We also found that the 2C protein has an enzymatic activity that converts AMP to a corresponding diphosphate using ADP or ATP as a phosphate donor. In addition, we observed that ATP hydrolysis results in 2C autophosphorylation. These findings indicate that the parechovirus 2C protein has enzymatic activities, which may contribute to several functions in the viral replication cycle. PMID- 16379009 TI - Nucleocytoplasmic shuttling of bovine herpesvirus 1 UL47 protein in infected cells. AB - Previous studies with transfected cells have shown that the herpes simplex virus type 1 (HSV-1) and bovine herpesvirus 1 (BHV-1) UL47 proteins shuttle between the nucleus and the cytoplasm. HSV-1 UL47 has also been shown to bind RNA. Here we examine the BHV-1 UL47 protein in infected cells using a green fluorescent protein-UL47-expressing virus. We show that UL47 is detected in the nucleus early in infection. We use fluorescence loss in photobleaching to show that nuclear UL47 undergoes rapid nucleocytoplasmic shuttling. Furthermore, we demonstrate that actinomycin D inhibits the reaccumulation of UL47 in the nuclei of infected cells. These results suggest that UL47 exhibits behavior similar to that of previously characterized RNA-transporting proteins. PMID- 16379011 TI - Tsunami donations help worldwide. PMID- 16379012 TI - A point mutation in NEMO associated with anhidrotic ectodermal dysplasia with immunodeficiency pathology results in destabilization of the oligomer and reduces lipopolysaccharide- and tumor necrosis factor-mediated NF-kappa B activation. AB - The NEMO (NF-kappaB essential modulator) protein plays a crucial role in the canonical NF-kappaB pathway as the regulatory component of the IKK (IkappaB kinase) complex. The human disease anhidrotic ectodermal dysplasia with immunodeficiency (EDA-ID) has been recently linked to mutations in NEMO. We investigated the effect of an alanine to glycine substitution found in the NEMO polypeptide of an EDA-ID patient. This pathogenic mutation is located within the minimal oligomerization domain of the protein, which is required for the IKK activation in response to diverse stimuli. The mutation does not dramatically change the native-like state of the trimer, but temperature-induced unfolding studied by circular dichroism showed that it leads to an important loss in the oligomer stability. Furthermore, fluorescence studies showed that the tyrosine located in the adjacent zinc finger domain, which is possibly required for NEMO ubiquitination, exhibits an alteration in its spectral properties. This is probably due to a conformational change of this domain, providing evidence for a close interaction between the oligomerization domain and the zinc finger. In addition, functional complementation assays using NEMO-deficient pre-B and T lymphocytes showed that the pathogenic mutation reduced TNF-alpha and LPS-induced NF-kappaB activation by altering the assembly of the IKK complex. Altogether, our findings provide understanding as to how a single point mutation in NEMO leads to the observed EDA-ID phenotype in relation to the NEMO-dependent mechanism of IKK activation. PMID- 16379013 TI - Formation of the CRS2-CAF2 group II intron splicing complex is mediated by a 22 amino acid motif in the COOH-terminal region of CAF2. AB - CRS2-associated factors 1 and 2 (CAF1 and CAF2) are closely related proteins that function in concert with chloroplast RNA splicing 2 (CRS2) to promote the splicing of specific sets of group II introns in maize chloroplasts. The CRS2-CAF complexes bind tightly to their cognate group II introns in vivo, with the CAF subunit determining the intron specificity of the complex. In this work we show that the CRS2-CAF complexes are stable in the absence of their intron targets and that CRS2 binds a 22 amino acid motif in the COOH-terminal region of CAF2 that is conserved in CAF1. Yeast two-hybrid assays and co-fractionation studies using recombinant proteins show that this motif is both necessary and sufficient to bind CRS2. The 22-amino acid motif is predicted to form an amphipathic helix whose hydrophobic surface is conserved between CAF1 and CAF2. We propose that this surface binds the hydrophobic patch on the surface of CRS2 previously shown to be necessary for the interaction between CRS2 and CAF2. PMID- 16379014 TI - Left main coronary artery disease: is CABG still the gold standard? AB - Severe stenosis of the left main coronary artery (LMCA) is a coronary artery disease manifestation of critical prognostic importance. As a consequence of the survival advantage conferred by coronary artery bypass grafting (CABG) over medical therapy, lesions in the LMCA have been considered a standard indication for CABG for nearly 3 decades. Initial attempts to treat LMCA disease percutaneously by balloon angioplasty resulted in poor clinical outcomes, leading many to regard significant LMCA disease as a contraindication for percutaneous coronary intervention (PCI). However, the development and refinement of coronary stenting over the last 15 years, followed by the recent introduction of drug eluting stents, has fueled renewed interest in percutaneous treatment of LMCA disease. Outcomes of recent studies using sirolimus- and/or paclitaxel-eluting stents for treatment of LMCA disease have yielded rates of in-hospital and 1-year mortality that compare favorably with those of surgery. This article will review the natural history of LMCA disease, the outcomes of CABG for LMCA disease, and the history and recent developments regarding PCI for LMCA disease. PMID- 16379015 TI - Cardiovascular risk factors in the metabolic syndrome: impact of insulin resistance on lipids, hypertension, and the development of diabetes and cardiac events. AB - Metabolic syndrome (MS) is associated with excess cardiovascular risk above and beyond the contribution of traditional risk factors. It is a proinflammatory and prothrombotic condition associated with underlying insulin resistance. Hypertension and hyperlipidemia in the setting of MS are also associated with excess cardiovascular risk, as is the development of new onset diabetes during the course of therapy. Although impaired fasting glucose and impaired glucose tolerance (IGT) both predict the development of diabetes mellitus, IGT more strongly predicts CV events because it is associated with a greater degree of insulin resistance. Early recognition and aggressive lifestyle interventions are the cornerstones of treatment, with aggressive pharmacologic therapy introduced when appropriate. It is expected that future studies will more clearly define the early use of insulin-sensitizing agents in MS. PMID- 16379016 TI - Should angiotensin receptor blockers be added to angiotensin-converting enzyme inhibitors in the treatment of heart failure? AB - Angiotensin-converting enzyme (ACE) inhibitors have been the cornerstone of treatment of heart failure. Angiotensin receptor blockers (ARBs) remain an attractive alternative in heart failure patients intolerant of ACE inhibitors. The addition of ARBs to ACE inhibitors in the context of stable heart failure may lead to additional clinical benefits. This is in contrast to heart failure complicating acute myocardial infarction, in which it does not offer any therapeutic advantage. PMID- 16379018 TI - Coronary artery disease in patients with Buerger's disease. AB - Buerger's disease (thromboangiitis obliterans) is a small- and medium-vessel vasculitis that predominantly involves the distal extremities. Coronary involvement in Buerger's disease is very unusual and rarely reported. We describe the unusual coronary angiogram findings of a 61-year-old Caucasian male with Buerger's disease who underwent cardiac catheterization for preoperative cardiac risk stratification prior to a carotid endarterectomy. Following the case presentation, we review the pathology, pathogenesis, and diagnostic criteria for Buerger's disease, and also include a review of the existing literature on coronary involvement in Buerger's disease. PMID- 16379019 TI - Myocardial infarction. Initial results from the EPHESUS trial. PMID- 16379020 TI - Coronary artery disease. Randomized trials, registries, and revascularization. PMID- 16379021 TI - Expressed sequence tag analysis of zebrafish eye tissues for NEIBank. AB - PURPOSE: To characterize gene expression patterns in various tissues of the zebrafish (Danio rerio) eye and identify zebrafish orthologs of human genes by expressed sequence tag (EST) analysis for NEIBank. METHODS: mRNA was extracted from adult zebrafish eye tissues, including lenses, anterior segments (minus lens), retinas, posterior segments lacking retinas, and whole eyes. Five different cDNA libraries were constructed in the pCMVSport6 vector. Approximately 4,000 clones from each library were sequenced and analyzed using various bioinformatics programs. RESULTS: The analysis yielded approximately 2,500 different gene clusters for each library. Combining data from the five libraries produced 10,392 unique gene clusters. GenBank accession numbers were identified for 37.6% (3,906) of the total gene clusters in the combined libraries and approximately 50% were linked to Unigene clusters in the current database. Several new crystallin genes, including two gammaN-crystallins, and a second major intrinsic protein (MIP) were identified in the lens library. In addition, a zebrafish homolog of cochlin (COCH), a gene that may play a role in the pathogenesis of human glaucoma, was identified in the anterior segment library. Surprisingly, no clear ortholog of the major retinal transcription factor Nrl was identified. CONCLUSIONS: The zebrafish eye tissue cDNA libraries are a useful resource for comparative gene expression analysis. These libraries will complement the cDNA libraries made for the Zebrafish Gene Collection (ZGC) and provide an additional source for gene identification and characterization in the vertebrate eye. PMID- 16379022 TI - MMP-2 expression in uveal melanoma: differential activation status dictated by the cellular environment. AB - PURPOSE: MMPs are recognized to play a major role in tumor progression and metastasis of many forms of cancers. The purpose of this study was to compare the expression and activity of MMP-2 in uveal melanoma cell lines grown either in vitro on plastic culture plates or in vivo as tumors produced in chick embryos. METHODS: The chick chorioallantoic membrane (CAM) model was used to evaluate the tumorigenic potential of uveal melanoma cell lines derived either from the primary uveal melanoma tumor isolated from three different patients (cell lines SP6.5, SP8.0, and TP31) or from a metastatic lesion derived from the liver of a patient diagnosed with uveal melanoma (cell line H79). The presence of MMP-2 in the vicinity of the tumor cells was determined by immunofluorescence analyses. Gelatin zymography was used for the detection of latent and activated forms of MMP-2 in uveal melanoma cell lines when grown in vitro on plastic, or in the solid tumors these cell lines produced in vivo on the CAM of the chick embryo. The gelatinase activity was quantified by densitometric analyses and the active/(active+pro-form) ratio was calculated as the MMP-2 activation ratio. Western blot analyses were performed to confirm the zymographic profile. RESULTS: Only the inactive form of MMP-2 was expressed and secreted in vitro by all uveal melanoma cell lines, higher levels being found for the liver-derived H79 cell line whereas SP8.0 only expressed MMP-2 to a very low level. On the other hand, all solid tumors produced in the CAM from these cell lines expressed and secreted, although to varying levels (SP6.5 and SP8.0, TP31 and H79), primarily the active form of MMP-2. Gelatinolytic activities of active MMP-2 were significantly higher in uveal melanoma tissues than in the non-neoplastic CAM, as revealed by the measurement of the activation ratio. The immunolocalization of MMP-2 revealed that all cell lines were MMP-2-positive although a reduced and more diffuse staining was observed for H79 and SP6.5 than in SP8.0 and TP31 cells. CONCLUSIONS: These results suggest the activation of proMMP-2 as an important event in the process of uveal melanoma progression. An elevated active to inactive MMP-2 ratio in the tumor environment of uveal melanoma suggests that a potential MMP-2 activity could be related to the progression of this type of cancer. PMID- 16379023 TI - The effect of C3 transgene expression on actin and cellular adhesions in cultured human trabecular meshwork cells and on outflow facility in organ cultured monkey eyes. AB - PURPOSE: To determine the effects of adenovirus-delivered exoenzyme C3 transferase (C3) gene expression on cultured human trabecular meshwork (HTM) cells and on outflow facility in organ cultured monkey anterior segments. METHODS: An adenoviral (Ad) vector expressing both C3 and green fluorescent protein (GFP) was used to transduce cultured HTM cells. Changes in cell morphology and the organization of actin, vinculin, and beta-catenin were assessed using immunofluorescence. Cultured monkey eye anterior segments were used to test the effects of AdC3GFP on outflow facility. RESULTS: Treatment of HTM cells with AdC3GFP resulted in dose-dependent morphological changes 3 or 4 days post-transduction. The AdC3GFP-transduced cells were either partially retracted, rounded, or very elongated compared to non-transduced cells. Compared to AdGFP-transduced cells, AdC3GFP-transduced cells demonstrated disrupted actin cytoskeleton, reduced vinculin-positive focal adhesions, and loss of beta-catenin staining. Cells transduced with AdGFP did not round up or retract. In organ culture studies, outflow facility was increased by 90+/-21% (n=15, p<0.001) in AdC3GFP-transduced eyes compared to baseline and corrected for AdGFP-transduced control eye washout on days 3-6 after transduction. CONCLUSIONS: C3 transduction is effective in disrupting actin filaments, cytoskeleton, and cellular adhesions in HTM cells and in increasing outflow facility in organ cultured monkey anterior segments, suggesting that expressing the C3 gene in the trabecular meshwork may be an effective approach for glaucoma therapy. PMID- 16379024 TI - Examining the proteins of functional retinal lipofuscin using proteomic analysis as a guide for understanding its origin. AB - PURPOSE: To elucidate the origins of biologically active retinal lipofuscin (RLF) by examining its protein composition. METHODS: Total protein and total lipid were extracted and quantified. Proteins in this lipoprotein granule were identified by limited-scale proteomic analysis using both two-dimensional (2D) gel electrophoresis and SDS-PAGE coupled with MALDI-QqToF MSMS and automated LCMSMS, respectively. RESULTS: RLF granules were 44% protein and 50% lipid. Proteomic analyses identified 41 constituent proteins. Hydrophobic proteins and several proteins specific to photoreceptors, including rhodopsin, that have not previously been reported, were identified. Extensive protein modification, especially oxidative damage, was observed. CONCLUSIONS: Proteins identified support the model that RLF accumulates in RPE cells as a result of the buildup of undigested material from the phagocytosis of photoreceptor outer segments. Perhaps oxidative damage renders some of these proteins indigestible and thus leads to the accumulation of RLF granules. PMID- 16379025 TI - Y402H complement factor H polymorphism associated with exudative age-related macular degeneration in the French population. AB - PURPOSE: Identification of genetic factors for age-related macular degeneration (AMD) is of crucial importance in this common cause of blindness. A positive association between Y402H polymorphism of the complement factor H (CFH) gene and AMD has been recently reported in North American populations but not yet in European populations. The exudative form of AMD is rapidly progressive and usually associated with a severe prognosis. Our purpose was to investigate this association in a French population specifically affected with exudative AMD, in a case-control study. METHODS: Two series of unrelated exudative AMD patients, sporadic cases (n=60, mean age 74.9+/-5.7) and familial cases (n=81, mean age 74.0+/-9.4) were compared with healthy controls (n=91, mean age 74.6+/-6.3). The coding region of exon 9 of CFH was examined for the Y402H polymorphism by PCR direct sequencing. RESULTS: The 1,279-C allele frequencies were significantly higher in exudative AMD patients than controls (0.564 compared to 0.302; p<0.0001). Genotypic distribution of the Y402H polymorphism was significantly different between sporadic cases compared to controls (chi2=14.48 with 2 df, p<0.0007) and between familial cases compared to controls (chi2=23.78 with 2 df, p<0.0001). The odds ratio (OR) for exudative AMD was 3.00 CI95% (1.60-5.62) for heterozygotes (CT) and 6.93 CI95% (3.11-15.46) for homozygotes (CC). CONCLUSIONS: These results suggest the contribution of the Y402H polymorphism of the CFH gene to exudative AMD susceptibility also in the French population. This relationship with the CFH may lead to early detection and new strategies for prevention and treatment of AMD. PMID- 16379026 TI - Disease-associated mutations in CNGB3 produce gain of function alterations in cone cyclic nucleotide-gated channels. AB - PURPOSE: To characterize the functional consequences of disease-associated mutations in the CNGB3 (B3) subunit of human cone photoreceptor cyclic nucleotide gated channels in order to gain insight into disease mechanisms. METHODS: Three separate disease-associated mutations were generated in CNGB3: F525N, R403Q, and T383fsX. These mutant subunits were then heterologously expressed in Xenopus oocytes in combination with wild type CNGA3 (A3) subunits, and characterized by patch-clamp recording in the inside-out configuration. RESULTS: Co-expression of A3 and B3F525N, A3 and B3R403Q, or A3 and B3R403Q and B3T383fsX subunits resulted in channels that exhibited an increase in ligand sensitivity without a reduction in current density compared to wild-type heteromeric channels. Each simulated disease state produced channels that exhibited greater sensitivity to block by L cis-diltiazem than homomeric CNGA3 channels, confirming that the mutant CNGB3 subunits were competent to form functional heteromeric channels. Each combination of subunits displayed an increase in apparent affinity for cGMP relative to wild type heteromeric channels. However, F525N enhanced cGMP apparent affinity to a significantly greater extent than the other two modeled disease states. CONCLUSIONS: We have examined the gating effects of two previously uncharacterized disease-associated mutations in the CNGB3 subunit and found that in each case, the mutations resulted in a gain of function molecular phenotype. Furthermore, the magnitude of the effect on channel function correlated with the severity of the associated disease. The complete achromatopsia-associated F525N mutation resulted in more pronounced alterations in channel function than the mutation combinations linked to macular degeneration or progressive cone dystrophy. PMID- 16379027 TI - RPE65 surface epitopes, protein interactions, and expression in rod- and cone dominant species. AB - PURPOSE: RPE65 is an abundant protein necessary for the synthesis of the chromophore 11-cis retinal by the retinal pigment epithelium (RPE). Our purpose was to identify RPE65 surface epitopes, to assess protein interactions, and to evaluate RPE65 expression in eyes from rod- and cone-dominant species using a monoclonal antibody approach. METHODS: RPE65-specific monoclonal antibodies, mAb 8B11, and mAb 1F9, were generated using bovine RPE microsomal membranes and a human RPE65 synthetic peptide as immunogen, respectively. Western analysis was performed on bovine RPE membranes, as well as yeast strains generated by transfection with RPE65 cDNAs. Competition of antibody binding by synthetic peptides was assayed using ELISAs, western analysis, and elution from immunoaffinity matrices. RPE65 structural models were generated by ab initio and comparative methods. Immunohistochemistry was performed on retina/RPE/choroid cryosections and retina flatmounts. RESULTS: The antigenic determinant recognized by mAb 8B11 was localized to a 10 amino acid sequence, KVNPETLETI, that competed binding with microM affinity and eluted RPE65 from an immunoaffinity matrix incubated with solubilized bovine RPE membranes or RPE65-transfected cells. Similarly, solubilized RPE65 was bound and eluted from an mAb 1F9 immunoaffinity matrix using the immunizing peptide, FHHINTYEDNGFLIV. In both cases, 11-cis retinol dehydrogenase, but not other known visual cycle proteins, appeared to co elute with RPE65 in substoichiometric amounts. Both sequences localized to surface exposed regions of predicted RPE65 tertiary structures. RPE65 immunoreactivity was detected by mAb 8B11 and mAb 1F9 in the RPE, but not in retina, in bovine, rat, mouse, human, chicken, and Xenopus laevis, and in Nrl knockout mice whose retinas contain exclusively cone-like photoreceptor cells. CONCLUSIONS: The identification of RPE65 surface exposed antigenic determinants represents a first step toward understanding RPE65 structure and its interaction with visual cycle proteins, and provides a means for the purification of the native protein. The finding that RPE65 immunoreactivity is present in the RPE and not retina of both rod- and cone-dominant species does not support a proposed direct role for RPE65 in cone cell function. PMID- 16379028 TI - Normal HLA class I, II, and MICA gene distribution in uveal melanoma. AB - PURPOSE: The molecules of the HLA class I and II molecules as well as the MHC class I chain-related gene A (MICA), a polymorphic and stress-induced cell surface molecule, are involved in T-cell and natural killer-cell (NK-cell) mediated immune responses. In this study we looked for any genetic susceptibility contributed by HLA class I, class II, or MICA genes with regard to the development of uveal melanoma. METHODS: Between 1998 and 2001, 159 uveal melanoma patients were typed for HLA class I and II, and 168 uveal melanoma patients were evaluated for MICA by microsatellite typing. The HLA antigen and MICA allele frequencies were compared with control groups of, respectively, 2,440 and 247 healthy Dutch individuals. RESULTS: HLA class I, HLA class II, and MICA gene frequencies in uveal melanoma patients and healthy Dutch controls showed no significant deviations after correction for the number of comparisons. CONCLUSIONS: We conclude that there is no genetic susceptibility or increased risk attributed to any HLA class I, class II, and MICA polymorphism with regard to the development of uveal melanoma. PMID- 16379029 TI - Serum endostatin levels are elevated and correlate with serum vascular endothelial growth factor levels in patients with pituitary adenomas. AB - Endostatin, a cleaved fragment of collagen XVIII, is a potent endogenous angiogenesis inhibitor. Elevated serum endostatin levels have been recently reported in patients with various types of neoplasms. The purpose of our study was to evaluate serum concentrations of endostatin in patients harbouring various pituitary adenoma types and to examine the relationship of serum endostatin levels to circulating vascular endothelial growth factor (VEGF) levels. Preoperative serum endostatin and VEGF concentrations were measured using competitive enzyme immunoassays in 71 patients with pituitary adenomas (20 somatotropinomas, 3 corticotropinomas, 6 prolactinomas and 42 clinically nonfunctioning pituitary adenomas - CNFPAs) and compared with levels from age matched controls. In 35 patients postoperative immunohistochemical investigations were performed. Serum endostatin concentrations were significantly higher in all pituitary adenoma types, except for prolactinomas (somatotropinomas: 124 +/- 16; p < 0.02, corticotropinomas: 157 +/- 42; p < 0.02, prolactinomas: 141 +/- 37; p > 0.05, CNFPAs: 169 +/- 11 ng/ml; p < 0.000005 vs 73 +/- 10 ng/ml in controls). There was a significant positive correlation between endostatin and VEGF serum levels in patients with pituitary adenomas (r = +0.322; p = 0.006). In the control group a significant negative correlation xbetween circulating endostatin and VEGF was found (r = -0.653; p = 0.00975). The simultaneous elevation of endostatin and VEGF may attenuate the pro-angiogenic action of VEGF and be responsible for rather weak neovascularization of pituitary adenomas. Prospective studies are required to assess the usefulness of circulating endostatin and VEGF as markers of progression or recurrence of pituitary tumors. PMID- 16379030 TI - Effect of gsp oncogene on somatostatin receptor subtype 1 and 2 mRNA levels in GHRH-responsive GH3 cells. AB - Growth hormone releasing hormone (GHRH) signals via G protein-coupled receptors (GHRH-R) to enhance intracellular Galphas/adenylyl cyclase/cAMP signaling, which in turn has positive effects on GH synthesis and release, as well as proliferation of the GH-producing cells of the anterior pituitary gland. Some GH producing pituitary tumors express a constitutively active mutant form of Galphas (gsp oncogene). It has been reported that these tumors are more responsive to octreotide therapy. In this study we used a rat GH-producing cell line (GH3) stably transfected with the human GHRH-R cDNA (GH3-GHRHR cells) as a model to study the effects of gsp oncogene on somatostatin (SRIH) receptor subtype 1 and 2 (sst1 and sst2) mRNA levels. Transient transfection of gsp oncogene in GH3-GHRHR cells for 48 h increased intracellular cAMP levels and GH release. Phosphodiesterase (PDE) 4, sst1 and sst2 mRNA levels were increased by G protein mutation as assessed by real-time RT-PCR. Increased PDE mRNA levels in gsp transfected cells may be a compensatory mechanism to the constitutive activation of cAMP-dependent pathway by G protein mutation and is consistent with reports of higher PDE expression in human pituitary tumor that express gsp. Our data suggest that higher expression of sst1 and sst2 mRNA induced by the gsp oncogene may be a mechanism by which gsp-positive tumors show a greater response to SRIH. GH3 cells permanently transfected with GHRH-R can be used for in vitro studies of actions of GHRH. PMID- 16379031 TI - Stimulation of the hypothalamic-pituitary-adrenal axis with the opioid antagonist nalmefene. AB - Nalmefene Stimulation of the HPA Axis. BACKGROUND: The Hypothalamic-pituitary adrenal (HPA) axis plays a vital role in the body's response to stress. The traditional gold standard for evaluating the HPA axis, the insulin hypoglycemia test (IHT), has several known limitations, and a second test, the standard ACTH stimulation test, can detect severe deficiencies of cortisol, but often misses mild or early cases. Therefore, a better test for the evaluation of the HPA axis is needed. This study evaluated the opiate antagonist nalmefene as a stimulation test of the HPA axis. METHODS: 25 healthy subjects were studied, 9 women and 16 men, mean age 30.4 yr. (range 21-55), and mean BMI 24.1 kg/m2 (range 18.6-34.2). Subjects received one of 3 doses of intravenously administered nalmefene: 2 mg (n = 6), 6 mg (n = 12), or 10 mg (n = 7). Serum cortisol and plasma ACTH were measured before and serially over two hours after the administration of nalmefene. RESULTS: ACTH and cortisol levels rose significantly and similarly after the 10 mg dose and the 6 mg dose. After the 10 mg dose, mean peak ACTH was 82.4 +/- 22.6 pg/ml and mean peak cortisol was 25.2 +/- 1.8 microg/dl. After the 6 mg dose, mean peak ACTH was 70.3 +/- 7.7 pg/ml and mean peak cortisol was 24.7 +/- 1.7 microg/dl. Cortisol levels rose above 18 microg/dl in all subjects receiving 10 mg of nalmefene, and in all but two of the subjects receiving 6 mg of nalmefene. Side effects to nalmefene were of greater duration and intensity in the subjects receiving 10 mg of nalmefene vs. those receiving 6 or 2 mg. These included most notably fatigue, lightheadedness, nausea and vomiting. CONCLUSIONS: Of the nalmefene doses we studied, 6 mg achieved the best combination of stimulation of ACTH and cortisol and fewest side effects. If further studies show a concordance between nalmefene and IHT, nalmefene testing could be used to assess the HPA axis in patients at risk for dysfunction of this axis. PMID- 16379033 TI - Spontaneous gonadotrophin deficiency recovery in an adult patient with Langerhans cell histiocytosis (LCH). AB - Langerhans cell histocytosis (LCH) is a rare disease which exhibits a particular predilection for pituitary involvement leading to diabetes insipidus (DI) and other anterior pituitary hormonal deficiencies that are usually permanent and unresponsive to treatment. We report a 35 year old woman with a 10 year history of multisystemic LCH who developed DI, mild hyperprolactinemia, gonadotrophin and partial growth hormone deficiency following a normal delivery that was accompanied with infundibular thickening on pituitary magnetic resonance imaging (MRI). Following several courses of glucocorticoid administration, that were not associated with any substantial improvement, the patient was started on estrogen replacement therapy and cabergoline. After a three year period free of further relapses she developed irregular uterine bleeding. Following estrogen and cabergoline discontinuation she resumed normal menstruation while a repeated MRI of the pituitary showed an almost normal infundibulum. Endocrine investigation revealed normal gonadotrophin axis and prolactin levels, while the patient continues to menstruate, every 30-40 days, ten months after the resumption of her menstrual cycle. This case demonstrates for the first time that LCH induced pituitary deficiencies can run a variable clinical course and even spontaneously recover. PMID- 16379032 TI - Long-term efficacy of bromocriptine in macroprolactinomas and giant prolactinomas in men. AB - We prospectively analyzed presentations and long-term therapeutic responses to bromocriptine (BRC) in 29 newly diagnosed men with macroprolactinomas including 14 patients with 'giant prolactinoma'. Clinical symptoms, prolactin (PRL) levels and tumor size on MRI were measured before BRC and sequentially thereafter. The duration of follow-up were 6 to 96 (30.7 +/- 14.4) months. Pretreatment PRL ranged between 124 and 29200 ng/mL (1698 +/- 857.1) and tumor volume was between 2.81 and 132 cm(3) (21.1 +/- 24.3). Baseline PRL levels did not correlated with tumor volume (r = 0.45, P > 0.05). Significant decrease (P = 0.0003) in PRL, at least 96% of the pretreatment value from 1698 +/- 857.1 ng/mL to 42.4 +/- 30.6 ng/mL occurred in 26 patients. Persistent normalization of PRL levels (< 16 ng/mL) for at least 6 months was achieved in 12 patients (40.8%). Twenty-two patients (74.8%) achieved significant tumor shrinkage (P = 0.005) at study completion. An improvement in visual field defects (VFD) and restoration of libido and potency was observed in 40% and 33.3%, respectively. Trans-sphenoidal / trans-frontal pituitary surgery was performed in 9 patients (31%) for various reasons: pituitary apoplexy in 1, CSF rhinorrhea in 2, increasing prolactin in spite of BRC therapy in 3, and intolerant /resistant to BRC in 3 patients. These data suggest that, in male macro- and giant prolactinomas, dopamine agonists represent the first-line therapy effective in reducing PRL, restoration of libido and potency, improvement of VFD and determining tumor shrinkage. PMID- 16379034 TI - Lymphoma metastasizing to the pituitary: an unusual presentation of a treatable disease. AB - Lymphoma involving the pituitary gland is particularly rare. We present two cases of patients with pituitary lymphoma, both of whom were symptomatic from pituitary dysfunction. The first patient demonstrated pituitary involvement on imaging, with mild biochemical diabetes insipidus but clear hypoadrenalism. Both adrenals were grossly enlarged on CT scanning and biopsy of one of the adrenal masses confirmed the diagnosis of diffuse large B cell lymphoma. The second patient presented with clinical diabetes insipidus but with no obvious abnormalities on pituitary imaging. CT scanning of abdomen and pelvis, however, revealed widespread lymphadenopathy. Lymph node biopsy revealed a T cell-rich B cell lymphoma. Review of the English-language literature of all published cases of pituitary lymphoma in the presence of generalised disease in immunocompetent patients revealed 13 cases. Most patients had large B cell non-Hodgkin's lymphoma. Involvement of the anterior lobe of the pituitary was more frequently seen than in patients developing pituitary metastases from solid tumour primaries. Patients with advanced lymphoma including the pituitary also appear to have a better prognosis than patients presenting with pituitary metastases. This is an important diagnosis to make as rapidly as possible to allow the early institution of effective therapy. PMID- 16379035 TI - Serum IGF-1 in treated acromegaly - how normal is "normal"? AB - We describe four acromegalic patients with persisting typical symptoms - excessive sweating, lack of suppleness of hands, joint pains - despite the achievement of normal serum IGF-1 levels after pituitary surgery. In three patients there was a clear improvement in symptoms when lower IGF-1 levels within the normal range were achieved with pegvisomant treatment. In the fourth patient IGF-1 levels have fluctuated within the normal range with persistence of abnormal sweating, particularly at night. Two of three patients who had an oral glucose tolerance test when serum IGF-1 was in the normal range failed to suppress GH levels to less than 1 ng/ml. We conclude that, in the treated acromegalic patient, IGF-1 levels within the normal range need to be looked at critically to determine what is truly normal for that individual. Relief of symptoms seems a reasonable yardstick, in addition to population norms, by which to judge whether the prevailing IGF-1 level is appropriate; in some cases the aim should be an IGF 1 level in the lower half of the normal range, or perhaps even the lowest quartile. PMID- 16379036 TI - Clinical and molecular features of a TSH-secreting pituitary microadenoma. AB - We describe a case of a thyroid stimulating hormone (TSH)-secreting pituitary microadenoma, and report the systematic gene expression profile of the surgically removed tumor. A 50-year-old woman was referred to our hospital because she had high TSH, free-T4, and free-T3 levels, and a pituitary tumor that was visualized with magnetic resonance imaging. Her basal TSH level was high even after a high T3 loading dose, and increased following administration of thyroid releasing hormone (TRH) even after administration of a high dose of exogenous T3. Her clinical symptoms and peripheral markers for T3 were responsive to exogenous T3. There was no thyroid hormone receptor (TR) beta gene mutation. The patient was diagnosed with a TSH-secreting pituitary adenoma, and trans-sphenoid surgery was performed. The histologic features and immunophenotype were consistent with a TSH secreting pituitary adenoma. Reverse transcription-polymerase chain reaction analysis of pituitary hormones, pituitary-specific transcription factors, receptors, and transcriptional cofactors of clinical significance was performed on the removed tumor. The tumor expressed TSH, growth hormone, prolactin, alpha subunit, pituitary transcription factor-1 (pit-1) but not proopiomelanocortin (POMC), prophet of pit-1 (prop-1) and pituitary cell-restricted T box factor (Tpit). TRbeta and TRH-receptor gene expression was normal. Three steroid receptor coactivators (SRC)-1, SRC-2, and SRC-3 were expressed. Nuclear receptor corepressor (N-CoR)2 was absent in the tumor, whereas nuclear receptor corepressor (N-CoR1) was expressed. Somatostatin receptor type 1 expression was significantly decreased, whereas type 4 receptor was expressed, which are unusual characteristics for pituitary tumors. The gene expression pattern in the tumor might have a role in the clinical features of this case. PMID- 16379037 TI - Intracranial aggressive fibromatosis presenting as panhypopituitarism and diabetes insipidus. AB - Aggressive fibromatosis (AF) is a rare, locally aggressive, proliferative fibroblastic lesion affecting musculoaponeurotic structures, most often, of the limbs and trunk. Intracranial AF is extremely rare and requires aggressive treatment to prevent recurrence. We present a case of a 34 year-old male with AF involving intracranial structures causing panhypopituitarism and diabetes insipidus. Patient was admitted to hospital because of polyuria, polydipsia, and loss of libido, impotence, hearing loss, and gait disturbance. On cranial magnetic resonance imaging, the lesion extended through the sphenoid sinus into the both pterygoid recesses, destroying the left lateral wall of the sphenoid sinus and invading the retroorbital area. There was also a distinct lesion in the hypothalamic area. The tumor was markedly isointense on both T2- and T1-weighted images relative to gray matter, and enhanced strongly after administration of gadolinium. The patient underwent partial resection of the lesion via a transcranial approach. The pathological examination of the mass was reported as AF. No other sites were found to be involved by thorax and abdominal tomography. Hormonal assessment of hypothalamic-pituitary dysfunction revealed panhypopituitarism with central diabetes insipidus. Replacement therapy was instituted. In this case, standard treatment of wide-field surgical resection was impossible. On the basis of reports that radiotherapy is an effective treatment for this kind of tumor, we administered radiation to the affected area, since chemotherapy and hormonal treatment of non-resectable tumors are not satisfactory. To our knowledge, this is the first reported case of AF presenting as panhypopituitarism with central diabetes insipidus. PMID- 16379038 TI - A phase II open-label trial of apomine (SR-45023A) in patients with refractory melanoma. AB - Metastatic melanoma continues to be a very difficult disease to treat. Options are limited and often have very little impact on the course of the disease. The objective of the current study was to evaluate the efficacy and safety of continuously administered Apomine (SR-45023A), a novel bisphosphonate, in patients with previously treated metastatic malignant melanoma. Adult patients with previously treated metastatic melanoma received Apomine 100 mg orally, twice daily (total dose 200 mg per day) continuously for 28 days (defined as a cycle). Treatment was continued until disease progression or unacceptable toxicity. A total of 42 patients received at least one dose of Apomine. Stable disease was achieved in 2 patients (5%). No complete or partial responses were observed. Progression free survival of at least 16 weeks was observed in 6 patients (14%). The median overall survival was 6.1 months (95% CI, 4.9-9.4 months). Time to treatment failure was 1.7 months (95% CI, 1.6-1.8 months) with Apomine therapy. By cycle 2, Apomine concentrations reached steady-state. Apomine was well tolerated with only 37% of patients experiencing any drug-related event. Abdominal pain was the most frequent adverse event occurring in 26% of patients. In conclusion, Apomine, at the current dose studied, failed to produce a 30% progression free survival rate at 16 weeks considered to be a meaningful benefit for further development. PMID- 16379039 TI - Phase I clinical trial of weekly docetaxel and exisulind, a novel inducer of apoptosis. AB - BACKGROUND: The objective of this phase I study was to determine the maximal tolerated dose (MTD) of the combination of weekly docetaxel and exisulind in patients with advanced solid tumors. PATIENTS AND METHODS: Patients with advanced or refractory solid tumors were treated with intravenous weekly docetaxel with daily oral exisulind. The following dose levels (docetaxel/exisulind) were explored: 30-mg/m2/200 mg po bid, 35/200, 35/250 and 40/250. Docetaxel was administered weekly for 6 weeks followed by 2 weeks off, and exisulind was taken twice daily. Each cycle was 8 weeks. RESULTS: Eighteen patients were enrolled in the study. All of them had received prior systemic therapy. Most patients had either melanoma or carcinomas of the upper gastrointestinal tract. A total of 31 cycles of therapy were administered. DLTs were grade 3 diarrhea, anorexia and fatigue and grade 3 cutaneous toxicity at dose level 4 (40/250). Myelosuppression was mild. Fatigue and gastrointestinal toxicity (anorexia, dyspepsia, nausea, abdominal pain and diarrhea) represented the most common toxicities. However, grade 3 and grade 4 toxicities were uncommon. There were no treatment related deaths. No objective responses were observed and five patients achieved stable disease. CONCLUSIONS: The recommended dose for phase II studies is weekly docetaxel 35 mg/m2 for 6 weeks followed by 2 weeks off in combination with oral exisulind 250 mg po bid. This combination is feasible and well-tolerated at these doses. PMID- 16379040 TI - Effects of the tumor vasculature targeting agent NGR-TNF on the tumor microenvironment in murine lymphomas. AB - TNF-alpha may improve drug delivery to tumors by alteration of vascular permeability. However, toxicity precludes its systemic administration in patients. NGR-TNF comprises TNF coupled to the peptide CNGRC, which is a ligand for CD13. CD13 is expressed on tumor vasculature. Therefore, to assess the efficacy of NGR-TNF its biological effect on tumor vasculature should be measured rather than its effect on tumor growth. The aim of this study was to assess the effects of a low dose of NGR-TNF (5 ng/kg) on vascular permeability, tumor hypoxia, perfusion and proliferation in lymphoma bearing mice. MRI measurements with blood pool contrast agent showed an increased leakage of the contrast agent from the vasculature in NGR-TNF treated tumors compared with controls (p < 0.05), suggesting NGR-TNF-induced vascular permeability. Immunohistochemical analysis two hours after NGR-TNF treatment showed a decrease in tumor hypoxia (p < 0.1) and an increase in labeling index of the S-phase marker bromodeoxyuridine (p < 0.1), possibly due to an increase in tumor blood flow after NGR-TNF treatment. Although a decrease in tumor hypoxia and an increase in labeling index could have lead to increased tumor growth, in this experiment after one day a decrease in tumor volume was measured. Possibly, the effects on tumor hypoxia and proliferation two hours after treatment are transient and overruled by other, more longlasting effects. For example, the observed increase in vascular permeability may lead to haemoconcentration and increased interstitial pressure, ultimately resulting in an reduction of tumor blood flow and thus a decrease in tumor growth. A beneficial effect of NGR-TNF in combination with other therapeutical agents may therefore critically depend on the sequence and timing of the regimens. Currently, NGR-TNF is being tested in clinical studies. PMID- 16379041 TI - The combination of sulfinosine and 8-Cl-cAMP induces synergistic cell growth inhibition of the human neuroblastoma cell line in vitro. AB - To identify purine analogs that could be effective in treating neuroblastomas, we tested the anticancer properties of sulfinosine, 8-Cl-cAMP and 8-Cl-adenosine in the SK-N-SH cell line. First we examined the effects of these three agents on cell growth inhibition and cell viability by the BrdU and Sulforhodamine B assay. Treatment of SK-N-SH cells with increasing concentrations of these compounds led to a significant inhibition of cell proliferation and decrease of cell viability in a time- and dose-dependent manner at micromolar concentration (<10 microm). Treatment with a combination of sulfinosine and 8-Cl-cAMP resulted in synergistic effects on growth inhibition, cell cycle arrest and induction of apoptosis. Flow cytometric analysis showed that 8-Cl-cAMP arrested the cells in the G0/G1 phase and sulfinosine blocked cell cycle progression at the G2/M stage, in contrast to the combined effects of both agents that did not arrest growth at any particular phase of the cell cycle. Further analysis of apoptosis induction demonstrated an increase from 17 to 24% of both early and late apoptotic cells and a very low percentage of necrotic cells. These results indicate that apoptosis was the predominant type of cell death after treatment of SK-N-SH cells with both substances, as well as with their combinations. PMID- 16379043 TI - [The self healing power of cities--civil qualities of urban life]. AB - Health and hygiene are top issues from the early days down the history of a town or a city. Pestilence, epidemic plagues and infectious diseases are once in a while endangering the delicate equilibrium of a city as an organism. Many infra structural measures are taken to ensure health in and of the city. What makes a healthy city is not just the absence of illness, a healthy city is inspired by the mentality of their citizens, it is a work of art and based on an consensus of what does mean quality of live for the city and its inhabitants. It is multifaceted and can develop the power to regenerate itself. In times of a demographic change with more elderly people in the future then ever and less money in public pursues we nevertheless have the civic duties and the obligation to look after health of our cities. Many faculties like science, visual art, engineering and design must cooperate and work together toward that goal. PMID- 16379042 TI - In vitro characterization of the human biotransformation and CYP reaction phenotype of ET-743 (Yondelis, Trabectidin), a novel marine anti-cancer drug. AB - ET-743 is a potent marine anti-cancer drug and is currently being investigated in phase I and II clinical trials, e.g. in combination with other anti-cancer agents. To assess the biotransformation and CYP reaction phenotype and their potential implications for human pharmacology and toxicology, the in vitro metabolism of ET-743 was characterized using incubations with human liver preparations, cytochrome P450 (CYP) and uridine diphosphoglucuronosyl transferase (UGT) supersomes.CYP supersomes and liver microsomes showed that ET-743 was metabolized mainly by CYP3A4, but also by CYP2C9, 2C19, 2D6, and 2E1. ET-743 showed the highest affinity for CYP3A4 and the highest maximal metabolic rate for CYP2D6 among the CYPs shown to metabolize ET-743. In addition, the Km value of ET 743 in female microsomes was significantly lower compared to male microsomes, while the Vmax values did not differ. ET-743 glucuronidation, catalyzed by UGT2B15, was observed in microsomes and S9 fraction. In addition, conjugation by glutathione-S-transferase and no sulphation was observed for ET-743 in cytosol and S9 fraction. ET-743 was more extensively metabolized when CYP activity was combined with phase II enzymes UGT and glutathione-S-transferase (GST), indicating that CYP, UGT, and GST simultaneously metabolize ET-743 in the S9 fraction. These results provide evidence that CYP3A4 has a major role in the metabolism of ET-743 in vitro with additional involvement of CYP2C9, 2C19, 2D6, and 2E1. Furthermore, ET-743 is conjugated by UGT and GST. This information could be important for interpretation of the pharmacokinetic data of clinical trials and prediction of drug-drug interactions. PMID- 16379044 TI - [Senior migrants in Bremen: demographic structure, social condition and health status]. AB - In the coming years, the number of migrants of 60 and older will rise considerably in Germany. There are indications that their health problems are more severe than those of the native population, and that much can be done to supplement health care offers for them. The authors suggest to put this statement on a more objective basis and bridge the gaps of empirical research, focussing both on health relevant aspects of the general situation of senior migrants (differences in the cultural background, migration biography, accommodation, integration/segregation etc.) and on the health status itself. At the same time preconditions and possibilities of an intercultural opening of care structures and culture-sensitive nursing should be utilised and further developed. PMID- 16379045 TI - [Methods of infectious disease epidemiology for public health services: case control studies as bridge between practical work, surveillance and epidemiology]. AB - Routine surveillance of infectious diseases is one important task of national public health services. Due to globalization the impact of international disease monitoring rises. The volume of traffic and trade is constantly increasing, which makes a spread of infectious diseases to distant places more likely then ever. This are challenges international infectious disease networks have to deal with. Thus, the role of investigation of disease outbreaks is not only relevant for local level; it furthermore prevents broader spread. To eliminate outbreak sources good epidemiological analyses have to be conducted. However, this often leads to extra time and personal resources. In addition classical studies, like case-control studies, underlay methodical limitations. Case-control studies, especially if conducted on the basis of notified cases from surveillance data, are susceptible to bias (like selection- and recall bias). Another study design, the case-case study (case-case comparison), provides an alternative, which is less affected by such limitations. In this approach a number of cases related within an outbreak, is compared with a group of infected individuals of the same disease, not belonging to the outbreak. Results from recently published studies lead to the assumption that this study design is adequate for the investigation of infectious disease outbreaks, captured by surveillance systems. The practical application of this study design makes it especially useful for local public health services and thus provides a bridge between surveillance, epidemiology and practical work in Public Health. PMID- 16379046 TI - [Accumulated occurrence of illnesses with Salmonella enteritidis in hospitals and nursing homes in the district Oberallgaeu, Bavaria, in July 2004]. AB - We report on the investigation of a Salmonella enteritidis outbreak in hospitals and nursing homes in the district Oberallgaeu, Bavaria, in July 2004. Affected by this outbreak were hospital patients, inhabitants of nursing homes, kitchen coworkers and maintenance personnel. Within this outbreak six deaths were observed. A forensic medical investigation of four deaths revealed two cases of Salmonella enteritis as the primary cause of death, another cause of death was due to other causes and one cause of death remained unclear. The microbiological investigation of stool samples linked all positive samples to an outbreak during this period. The epidemiological outbreak curve pointed towards a foodborne transmission of the pathogen. The infection period could be limited to the weekend of the 3./4.7.2004. All affected institutions were supplied by a catering service with several sites. All tested food samples were negative for Salmonella enteritis. Epidemiological investigations revealed that a contaminated pudding was probably responsible for the outbreak. PMID- 16379047 TI - [Cluster of meningococcal diseases in Allgaeu -- strategies for intervention]. AB - In spring 2004 an accumulation of cases of invasive meningococcal disease was observed in the Allgaeu/Bavaria. Investigations of the isolates showed, that four cases in neighbouring municipalities of the district Oberallgaeu were caused by an identical strain of serogroup C. The particular strain was a rare variant of the so called ET-15 clone, which had caused several outbreaks of severe meningococcal disease among young people in the past, for example in Rottal/Inn (1998), Karlsruhe (1999 - 2000) and Schwerte (2003). The involved health authorities had to decide, which intervention strategies were reasonable and appropriate to the given situation. An epidemiological assessment of the situation was made by the Bavarian Health and Food Safety Authority (LGL) using the recommendations of the permanent immunization committee at the Robert Koch Institute and of the Advisory Committee on Immunization Practices of the Centers for Disease Control and Prevention (CDC) . The LGL together with the Robert Koch Institute and the national meningococcal reference centre concluded that the situation fulfilled the criteria for a vaccination indication in accordance with section sign 20 Abs. 5 of the infectious disease control act (Infektionsschutzgesetz, IfSG). On the basis of this assessment the responsible regional health authority issued a public recommendation for vaccination and the district health authority of the Oberallgaeu was assigned to implement a vaccination campaign. The Oberallgaeu health authority offered vaccination sessions to the public in the concerned communities. The target group comprised babies, children, young people and adults up to twenty years, who lived in the concerned communities in the northern part of Oberallgaeu, as well as close contacts of cases and members of the above age group, who had visited communal facilities in the communities concerned. Our report describes the implementation of the vaccination campaign. PMID- 16379048 TI - [Risk factors for abundance of shiga toxin producing Escherichia coli in sewage water]. AB - Shiga toxin producing Escherichia coli (STEC) in sewage influent into surface water are a potential source of human infections with STEC. Eight sewage treatment plants in Bavaria, Germany, were sampled at regular intervals from 2003 to 2004 in order to estimate STEC load and quantify risk factors. 95 of 378 samples (25 %) were tested positive for stx1and/or stx2 with PCR after enrichment culture. STEC elimination after treatment was 44 %. The following risk factors were analysed with logistic regression: location of sewage plant (rural vs. urban), treatment plant technology (two stage vs. three stage treatment) and sampling location (sewage input vs. sewage output). Rural plants had odds-ratios of 1,7 (95 % CI 1.03 - 2.69; p = 0.038) for a positive stx1 and/ or stx2 PCR result, sampling at sewage input of 2.1 (95 %CI 1.28 - 3.36; p = 0.003) and three stage plants of 1.51 (95 % CI 0.94 - 2.44; p = 0.087, not significant). Sampling after rain and after dry spells had no impact on STEC abundance (univariate Chi square test = 0.01; df1; p = 0.920). Rural sewage plants had higher odds of STEC content. The influence of the sewage plant technology on the STEC load requires further clarification. PMID- 16379049 TI - [Gender-specific health reporting in the Bodenseekreis: future theme of " health in boys and men"]. AB - Population prognoses predict a far over-proportional increase in the number of male seniors, entailing substantial implications for the planning of health supply and care, as well as various possibilities to intervene preventively by specific risk avoidance or boosting health responsibility. In the future men's health and male-specific health reporting will become eminently important. A gender-specific life table analysis for the years 1998 - 2002 shows which causes of death are more responsible for loss of life expectancy of men compared to women in both the Bodenseekreis and Baden-Wurttemberg. Baden-Wurttemberg and the Bodenseekreis are regions with a well above-average life expectancy compared to other regions in Germany. The average life expectancy in Baden-Wurttemberg in the year 2000 amounted to 82 years (Bodenseekreis: 82.2) in women, and 76.4 years (Bodenseekreis: 77.3) in men. Among the main causes for the 5.6 year difference in life expectancy are cardiovascular diseases, cancer illnesses, traffic accidents and suicide. These groups of causes of death explain the major part of the difference in average life expectancy. An analysis of death risks specific to age and gender shows some promising approaches for health promotion and prevention based on identification of critical phases of life. In this case, special attention should be paid to infancy, as examinations made within the scope of enrolment at schools identify considerable differences in the prevalence of various disturbances. However, early adulthood and old age also prove to be important periods concerning men-specific interventions. The presented results show first possible starting points which, however, should above all make a contribution to the establishment of the topic of men's health as a focus on gender-specific health reporting, health promotion and prevention. The described target group for health promotion and prevention is, on the one hand, sufficiently large, on the other hand, sufficiently disadvantaged in terms of health to demand more attention in the future. PMID- 16379050 TI - [Psychiatric significance of stalking]. AB - AIMS: To provide an overview on the psychiatric significance of stalking and the implications for the health care system. RESULTS: Stalking is a widespread phenomenon and can result in significant health care problems. CONCLUSION: In the future physicians will have to deal with this problem in a professional manner. PMID- 16379051 TI - [Results of two nationwide "smoke-free" campaigns]. AB - BACKGROUND: "Rauchfrei" -- the German part of the international contest "Quit and Win"-- is the biggest smoking cessation campaign in Germany. Already in the year 2000 24,925 smokers participated. A total of 90,458 adolescents and adults participated in "Rauchfrei 2002", that means about 0.5 % of all German smokers. OBJECTIVE: Main aim of the quit smoking campaign is to motivate as many smokers as possible to stop smoking for at least four-weeks and herewith to achieve a stop of the tobacco consumption in the long run. In this context the continuous 4 weeks and 12-months abstinence rates of the participants serve as indicators for success. METHODS: A survey with the adult participants aged 18 and older was carried out one year after each campaign. The samples were drawn randomly from all 10 German postal zip code-areas. A response rate of over 60 % in both surveys resulted in a sample size of n = 651 (2000) and n = 996 (2002). RESULTS: By the end of "Quit and Win 2000" (2002) 72 % (69 %) of the participants were smoke free; 12 months after the campaign 30 % (22 %) of the participants were continuously smoke-free. Overall, men were more successful than women, and participants with a lower education were as successful as participants with a higher school education. Substantially more participants, who had received support from either family members, friends or colleagues, had abstinence rates above the average. CONCLUSION: "Rauchfrei" was successfully implemented in Germany and contributed to draw attention to the risks of smoking nationwide. PMID- 16379052 TI - [What significance does the topic 'prevention' have in social-psychiatric research? A systematic literature analysis]. AB - AIM OF THE STUDY: Scientists and politicians currently pin great hopes on preventive activities in order to increase the health of particular target populations and to reduce illness related costs. This also holds true for mental disorders, which often show a chronic course of disease. The aim of the present study is to analyse the importance of prevention in social-psychiatric research. METHODS: Including 60 journals, listed in the Science Citation Index or the Social Science Citation Index for the psychosocial field, a systematic literature analysis was done for the year 2004. RESULTS: Only 14 percent of social psychiatric original research paid attention to prevention. 39 percent of the prevention related publications refer to primary prevention, further 39 percent to secondary prevention, 22 percent focused on tertiary prevention. Research activities concentrated on prevention of substance related disorders as well as early detection or rehabilitation of schizophrenia. CONCLUSIONS: Social psychiatry has only partly responded to the growing importance of prevention in health care. Because many social-psychiatric questions are important for prevention research, social psychiatry should use their competences to facilitate prevention-studies. This holds particularly true for depressive disorders and dementia, which are conditions with high prevalence and substantial illness related costs. PMID- 16379054 TI - [Beta receptor blockers in patients with obstructive respiratory diseases -- risks and alternatives]. AB - The use of beta-blockers should be avoided in the treatment of patients with asthma and COPD because of their broncho-constrictive effect if there are medical alternatives available. This is the case in arterial hypertension, glaucoma, supraventricular and ventricular dysrhythmia. The use of beta-I-selective drugs is indicated in the treatment of patients with coronary heart disease after myocardial infarction, since the benefit of reduction of morbidity and mortality through the coronary heart disease outweighs the potential risks if the patient is under close and careful observation and the initial dose of the beta-blocker is low. On principle patients suffering from asthma bronchiale should not be treated with beta-blockers because the risk of severe airway obstruction is higher than in patients with COPD. PMID- 16379055 TI - Repeat mediastinoscopy as a restaging procedure. AB - BACKGROUND: Repeat cervical mediastinoscopy is a diagnostic surgical procedure for preoperative nodal staging in patients with insufficient first mediastinoscopy, with recurrent or second primary lung neoplasms, and following induction chemotherapy or chemo-/radiotherapy for locally advanced lung cancer. The aim of this study was to critically analyse indications, technical characteristics, intra- and postoperative complications, also to define selection criteria for patients with a higher probability of successful complete resection. MATERIAL AND METHODS: 279 patients with lung cancer (66 female and 213 male patients, mean age 58 years, range 28 to 78 years) underwent repeat mediastinoscopy from 1968 to 2004, 12 because of inadequate first procedure (group A), 67 because of recurrent lung cancer (group B) 35 because of second primary lung cancer (group C), and 165 following induction chemo-/radiotherapy for IIIa and IIIb disease (group D). The interval between first and second procedure was 17 days (range, 12 - 38) in group A, 14 months (range, 5 - 29) in group B, 27 months (range, 19 - 124) in group C, and 132 days (range, 113 - 145) in group D. RESULTS: No intra- or postoperative deaths were observed, 7 patients developed minor complications. N2 or N3 disease was found in 3/12 patients of group A (25 %), in 17/67 patients of group B (25.4 %) and in 6/35 patients of group C (17.1 %). Of the 116 patients with N2, and 49 with N3 disease before induction treatment (group D), repeat mediastinoscopy showed 126 N0, 20 N2 and 14 N3 status. Because of the presence of inseparable adhesions repeat mediastinoscopy was not possible in 5 cases. Five-year survival for patients with persistent N2 in repeat mediastinoscopy was despite surgery only 5 %. CONCLUSION: Repeat mediastinoscopy is a safe explorative procedure for the restaging of patients with primary locally advanced, recurrent or second primary lung cancer. In patients after induction treatment it is, however, less sensitive than the primary mediastinoscopy because of adhesions and fibrotic tissue. Patients with persistent N2 or N3 disease in repeat mediastinoscopy have a poor survival so that the indication for surgery has to be taken into consideration very carefully. PMID- 16379056 TI - [Diffuse smooth muscle proliferation of the lung and severe pulmonary hypertension]. AB - A 50-year old female forest worker presented with the complaints of progressive exercise induced dyspnea and the history of a recent syncope. Right heart catheterisation revealed the diagnosis of severe precapillary pulmonary hypertension. Ventilation perfusion lung scan indicated the existence of multiple pulmonary emboli with high probability. However, pulmonary angiography did not confirm this finding. Computed tomography of the lung demonstrated a slight mosaic pattern of diffuse ground glass opacities. After video assisted thoracoscopy, the histological diagnosis of diffuse smooth muscle proliferation of the lung could be concluded from the removed lung tissue. Diffuse smooth muscle proliferation of the lung is a rarely diagnosed disease, however it should be considered in patients with suspected pulmonary embolism by ventilation perfusion scan but a normal finding in pulmonary angiography, who are suffering from otherwise not explained pulmonary hypertension. PMID- 16379057 TI - [Nocturnal long-term recording of breath sounds in patients with bronchial asthma -- a reasonable diagnostic option?]. AB - Alterations in normal breath sounds are an important indication of pathophysiological processes in the bronchial system and the lung. In the presence of non-specific symptoms like dyspnea, wheezing or cough while sleeping it seems reasonable to make a nocturnal long-term recording of breath sounds. Thus specific auscultation phenomena can be recorded qualitative and particularly quantitative in the course. A mobile recording system provides a non-invasive monitoring at home which is independent of vigilance. Furthermore it can be used for follow-up examinations of medical interventions. PMID- 16379058 TI - [The management of spontaneous pneumothorax]. AB - The optimum management of spontaneous pneumothorax (SPTX) includes medical and surgical approaches but remains somewhat controversial due to inconclusive data background both in the treatment of the acute episode and preventive strategies. Current consensus recommendations suggest, that small and asymptomatic pneumothoraces can be managed conservatively by observation. Large and symptomatic pneumothoraces will respond to simple aspiration or small size catheter (< 16 F) insertion in about (2)/ (3) of cases. Otherwise, and in secondary SPTX, particularly in the elderly patient, large size (> 20 F) chest drainage, optimally combined with thoracoscopy (pleuroscopy), is the treatment of choice allowing success rates up to 96 %. Preventive strategies are indicated after the first recurrence and when predisposing pleuropulmonary lesions as in secondary SPTX are evident. With efficacy as a primary endpoint, currently available data do not provide clear evidence-based preference criteria between medical thoracoscopic talc pleurodesis and surgical strategies (usually VATS based partial pleurectomy +/- parenchymal resection). Therefore selection of the appropriate procedure remains influenced by expert opinion. However simple talc poudrage under medical thoracoscopy has been shown to be highly effective, safe and cheap at a cost level virtually not exceeding that of large size chest drainage. PMID- 16379059 TI - [COPD 2005 -- an overview]. PMID- 16379060 TI - [Pneumococci -- epithelial interaction]. PMID- 16379061 TI - [Prevalence of respiratory symptoms and diseases in neighbours of large-scale farming in Northern Germany]. AB - BACKGROUND: Up to now potential health effects of environmental exposure to intensive livestock production facilities have not been thoroughly studied. The aim of the Lower Saxony Lung Study (NiLS) was to assess the prevalence of respiratory symptoms and disease in an area with intensive animal production facilities taking into account environmental exposure to emissions from these large-scale farms. METHODS: All 10 864 inhabitants (age 18 to 44 years) of four towns with a large number of intensive animal production facilities in the area were invited to answer a mail-in questionnaire. Of these, 6416 subjects were randomly selected for the clinical part of the study (specific IgE, lung function measurements, bronchial provocation with methacholine). Overall, 6937 subjects answered the questionnaire (68 %), 60 % took part in the clinical tests. RESULTS: The prevalence of allergic disease in the study population was lower than in urban citizens. Subjects with more than 12 animal houses within 500 m of their home had an increased odds ratio for wheezing without a cold (odds ratio 2.7; 95 % confidence interval 1.4 - 5.4). They also showed a significantly decreased FEV (1) (mean 0.26 l; 0.04 l - 0.48 l) as well as a trend for a lower Tiffeneau Index. CONCLUSIONS: In result, a large number of intensive animal production facilities might be associated with decreased lung function results in the immediate neighbours. Further studies are necessary to confirm our findings. PMID- 16379062 TI - [Architecture of Sanatoriums around 1900]. PMID- 16379063 TI - [Prof. Dr. med. Karl Wurm (1906 - 2005) in memoriam]. PMID- 16379066 TI - Quantum dots: DNA detectives. PMID- 16379068 TI - Asymmetric nanoparticles: tips on growing nanocrystals. PMID- 16379070 TI - Nanoionics: ion transport and electrochemical storage in confined systems. AB - The past two decades have shown that the exploration of properties on the nanoscale can lead to substantially new insights regarding fundamental issues, but also to novel technological perspectives. Simultaneously it became so fashionable to decorate activities with the prefix 'nano' that it has become devalued through overuse. Regardless of fashion and prejudice, this article shows that the crystallizing field of 'nanoionics' bears the conceptual and technological potential that justifies comparison with the well-acknowledged area of nanoelectronics. Demonstrating this potential implies both emphasizing the indispensability of electrochemical devices that rely on ion transport and complement the world of electronics, and working out the drastic impact of interfaces and size effects on mass transfer, transport and storage. The benefits for technology are expected to lie essentially in the field of room-temperature devices, and in particular in artificial self-sustaining structures to which both nanoelectronics and nanoionics might contribute synergistically. PMID- 16379073 TI - Single-quantum-dot-based DNA nanosensor. AB - Rapid and highly sensitive detection of DNA is critical in diagnosing genetic diseases. Conventional approaches often rely on cumbersome, semi-quantitative amplification of target DNA to improve detection sensitivity. In addition, most DNA detection systems (microarrays, for example), regardless of their need for target amplification, require separation of unhybridized DNA strands from hybridized stands immobilized on a solid substrate, and are thereby complicated by solution-surface binding kinetics. Here, we report an ultrasensitive nanosensor based on fluorescence resonance energy transfer (FRET) capable of detecting low concentrations of DNA in a separation-free format. This system uses quantum dots (QDs) linked to DNA probes to capture DNA targets. The target strand binds to a dye-labelled reporter strand thus forming a FRET donor-acceptor ensemble. The QD also functions as a concentrator that amplifies the target signal by confining several targets in a nanoscale domain. Unbound nanosensors produce near-zero background fluorescence, but on binding to even a small amount of target DNA (approximately 50 copies or less) they generate a very distinct FRET signal. A nanosensor-based oligonucleotide ligation assay has been demonstrated to successfully detect a point mutation typical of some ovarian tumours in clinical samples. PMID- 16379081 TI - Medicare quality cancer care demonstration--one year later. PMID- 16379080 TI - The evolution of vesicles from bulk lamellar gels. AB - The remarkable ability of phospholipids to generate vesicles and lamellar morphologies has been mimicked by amphiphilic block copolymers and both classes of amphiphiles have been extensively studied in bulk and in dilute solution. The most common vesicle preparation method involves diffusion of water into a lamellar gel. The transformation proceeds from a collection of water-poor planar lamellae, to a water-rich interconnected layer-phase and then a collection of closed lamellae (vesicles) that form a close-packed gel. This is demonstrated by scattering and microscopy with the evolution of the gel structure being driven by increasing curvature. Vesicles have been observed to form when the copolymer water system still has long-range order, with a discrete, clustered-vesicle structure leading to a turbid dispersion. Eventually, at very low concentration, the packed vesicles separate, generating very stable isotropic dispersions that are transparent. This phase sequence, involving a dispersion of close-packed vesicles, accounts for the formation of a narrow size distribution in these systems. PMID- 16379082 TI - Toddlers, teens, and test tubes. PMID- 16379083 TI - Seeking soft skills. PMID- 16379084 TI - Management of acute coronary syndromes. PMID- 16379085 TI - Management of acute coronary syndromes. PMID- 16379086 TI - Management of acute coronary syndromes. PMID- 16379087 TI - Management of acute coronary syndromes. PMID- 16379088 TI - Management of acute coronary syndromes. PMID- 16379089 TI - Primary biliary cirrhosis. PMID- 16379090 TI - Primary biliary cirrhosis. PMID- 16379091 TI - Academic-industrial relationships. PMID- 16379092 TI - A critique of the motivational analysis in wrongful conception cases. AB - Most states now recognize a tort cause of action for wrongful conception, typically resulting from a failed sterilization. States differ, however, in determining whether damages should be awarded for child-rearing expenses and what factors juries can consider in setting such damage awards. This Note argues that one commonly used factor, the parents' motivation for selecting sterilization, is irrelevant and leads to inequitable results. Since the right to use contraception is constitutionally protected, the choice to sterilize in order to avoid financial burdens associated with child-rearing should not be given preferential treatment to sterilizations motivated by concerns of genetic defects or for the mother's health. PMID- 16379093 TI - Medicare Part D program warrants further scrutiny. PMID- 16379094 TI - Re: Overuse of technology. PMID- 16379095 TI - Findings of scientific misconduct. PMID- 16379096 TI - Abstracts of the 58th Annual Meeting of the Gerontological Society of America, November 18-22, 2005, Orlando, Florida, USA. PMID- 16379097 TI - [Acute coronary syndrome]. PMID- 16379098 TI - [Morphological and functional characteristics of the culprit coronary artery in acute coronary syndrome]. AB - In the catheter laboratory, faced with a patient with unstable angina, the problem is not the diagnosis of epicardial coronary artery stenosis but rather the atherothrombotic complexity of the stenosis and the identification of other less stenotic lesions on angiographically normal or nearly normal segments. The authors describe two appearances with morphological analysis of the vulnerable or unstable plaque and functional analysis of coronary artery vasomotricity. Acute coronary syndromes due to a clearly stenotic atherothrombotic plaque on angiography pose no diagnostic problems. However, an acute coronary syndrome may be more difficult to assess when angiography shows only simple irregularities of the lumen or no lesions at all. In these cases, angiography is said to be normal and the coronary arteries are considered to be possibly "healthy". Evaluation of the vessels' vasomotricity is especially valuable in this context. Using pharmacological stress tests, endovascular ultrasonography is a complementary diagnostic tool which allows identification of the unstable atheromatous process or of a partial atherothrombosis masked by positive remodelling process. These tests may also indicate the optimal treatment. PMID- 16379099 TI - [Myocardial perfusion and contrast echocardiography. Application to acute coronary syndrome]. AB - After mastering epicardial revascularisation, the therapeutic management of acute coronary syndrome has progressively turned towards the preservation of the capillary microcirculation. In this respect, contrast echocardiography provides all the answers to the main questions which arise, and is available within a timeframe and place compatible with the management of such patients. The techniques for microbubble detection, quantification tools, and contrast agents are well optimised for routine use both for initial diagnosis as well as monitoring the therapeutic effectiveness. It is only the administrative authorisation which is currently lacking treatment. PMID- 16379100 TI - [Acute coronary syndromes and myocardial perfusion. Cardiac magnetic resonance imaging and ultrafast CT]. AB - Imaging myocardial perfusion is essential in the management of acute coronary syndromes without ST elevation (ACS ST-) confirming the diagnosis of coronary lesions and quantifying the myocardial ischaemia, an important factor in the stratification of coronary risk. In ACS ST-, perfusion imaging allows evaluation of myocardial viability, diagnosis of residual ischaemia and also the detection of no-reflow phenomena after reperfusion procedures. Although myocardial scintigraphy is the reference method in clinical practice, it has many limitations such as its spatial resolution, its irradiation, its attenuation artefacts, and also the fact that it does not visualise the coronary arteries. This has led to the rapid development of two new non-invasive imaging techniques: cardiac MRI and ultrafast CT. The major advantage of MRI is the possibility of associating analysis of myocardial perfusion with that of cardiac muscle function by investigating right and left ventricular function at rest and during myocardial ischaemia stress tests and by analysis of myocardial viability. More recently, ultrafast CT has been clinically validated for coronary imaging. However, analysis of myocardial perfusion and ventricular function by CT scan is still only at the research stage. PMID- 16379101 TI - [Management of infarction with ST elevation in the acute phase: the current guidelines]. AB - The 2002 and 2005 recommendations of the European Society of Cardiology for the management of ST-elevation myocardial infarction at the acute stage are reviewed. Primary angioplasty should, whenever possible, constitute the default strategy, but intravenous thrombolysis still has an important role, particularly in the first few hours following symptom onset. PMID- 16379102 TI - [Acute coronary syndromes without ST elevation. State-of-the-art, recommendations, and best practices]. AB - In the last decade, acute coronary syndromes without ST elevation (ACS ST-) have become a clinical entity encompassing previous technologies such as acute coronary insufficiency and non-Q wave myocardial infarction. The recognition of its risks and, above all, the development and adaptation of medical and interventional strategies have transformed the prognosis. Patients presenting with ACS ST- are not all identical and do not all have the same prognosis; based on this observation, a number of prognostic scores have been proposed which help decide whether to use drugs such as GP IIb/IIIa inhibitors or coronary angioplasty. Conservative and aggressive strategies are opposing choices but risk stratification allows the treating physician to take a safe decision in the majority of cases. Besides angioplasty and GP IIb/IIIa inhibitors, the basic treatment (aspirin, low molecular weight heparin, clopidogrel, betablockers) should of course be prescribed and its efficacy has contributed to a better outcome in these patients. The recommendations of the scientific societies have evolved with time and increased understanding, allowing an evidence-based approach to the management of ACS ST- patients. PMID- 16379103 TI - [Prescriptions following acute coronary syndrome]. AB - Each year in France, 150,000 to 180,000 new patients are the subject of prescriptions following acute coronary syndrome with or without ST segment elevation. There are two targets of the treatment, atherosclerosis, a diffuse, evolving trouble which, in this situation, is coming out of an unstable phase, and the myocardium, which has often been revascularised and has suffered deterioration of its contractile and electrophysiological characteristics to a greater or lesser extent. Prescriptions, based on proven factors and always centred on hygiene and dietary advice and the use of a combination of statins and aspirin, are adapted to suit the atherosclerotic and myocardial risk assessed for the individual patient. The prescription starts off the secondary preventive phase. It marks the first stage of the follow up, which is inevitable though of variable duration, for a disease which may evolve. It is the first step in the accompaniment of an attentive, informed patient whose confidence has been restored and who must now avoid falling into the double trap of not taking the treatment sufficiently seriously or of obsessively over-reacting. PMID- 16379104 TI - [Post-myocardial infarction cardiogenic shock and circulatory assistance]. AB - Post-myocardial infarction cardiogenic shock still carries a very poor prognosis despite the rapid recourse to effective methods of myocardial revascularisation. Circulatory assistance devices allow restoration of adequate haemodynamics with limitation of myocardial work. In the most severe cases, implantation of intra thoracic devices is associated with a 70% survival rate in the latest series, providing they are used early. However, in many cases, the essential problem is to stabilise the patient's haemodynamic status, sometimes even before myocardial revascularisation. In these situations, implantation of a peripheral femoro femoral extra corporeal circulation (ECMO: extra corporeal membrane oxygenation) re-establishes an appropriate cardiac output andprovides time to transfer the patient, to perform coronary revascularisation or to assess neurological status, before deciding on the indications for more complicated assist systems. This "bridge to bridge" concept avoids the risk of implanting complicated assist devices in cerebrally dead patients or in those with multi-organ failure beyond treatment. Conversely, it gives some patients with apparent contraindications to complicated assist systems or who are unable to benefit from these systems for geographical reasons, a chance to survive. In early cardiogenic shock, the ECMO which has a low rate of complications, could safely promote myocardial recovery. PMID- 16379105 TI - [Management of chest pain by the Emergency Ambulance Service: the DOLORES register]. AB - Pre-hospital management of chest pain is a difficult problem. The emergency doctor has to take triage decisions based on instantaneous data whereas the decisional rationale of the many pathologies concerned, including acute coronary syndromes, is often based on observation over several hours. There have been few studies of the efficacy of pre-hospital management of chest pain by an emergency ambulance service. Therefore, the DOLORES register was set up to assess this problem over a 6 month period by the emergency ambulance service of Necker Hospital in Paris. Between January and June 2004, the Necker emergency ambulance service was called out on 205 occasions for chest pain. Forty-three patients had acute coronary syndromes (ACS) with ST elevation. Of the remaining 162 patients, 32 stayed at home, 2 were admitted the following day by cardiologists for coronary angiography, 52 were admitted for observation to the emergency unit and 76 were admitted to the coronary care unit. In the latter two groups, the final diagnosis of ACS without ST elevation was retained in 11/52 and 57/76 patients respectively. Finally, 2 patients were admitted directly to the catheter laboratory. The clinical and paraclinical data noted by the emergency ambulance service and at hospital admission was concordant in all cases. Pre-hospital triage by the emergency ambulance service seems to be effective. These results require confirmation with a large scale study. PMID- 16379106 TI - [Echocardiography and mechanical complications of recent myocardial infarction]. AB - The constantly advancing technology of echocardiography and its widespread usage in the intensive care unit has made it a routine examination in patients with acute myocardial infarction. It has become the reference method for diagnosis and monitoring of certain complications such as pericardial effusion, intra ventricular thrombosis, ventricular aneurysm and mitral regurgitation. The echocardiographic description of these complications dates back to the 1980s during which prospective studies accurately described the principal abnormalities. These descriptions have not been much improved upon with the advent of new technology. On the other hand, the frequency of these complications assessed in an era when reperfusion by thrombolysis or primary angioplasty was much less common than today, has considerably decreased. PMID- 16379107 TI - [Value of biochemistry performed in pre-hospital cardiology]. AB - For a long time, the diagnosis of an acute myocardial infarction (AMI) seen in outdoor patients, was only relying on ECG findings. For that reason a certain amount of patients suffering from an AMI showing an atypical or not contributive ECG had not been identified as such and in consequence did not benefit from any prehospital treatment or had not been admitted in coronary care unit (CCU). With the arrival of the biological bed side monitoring in the SAMU, it became possible to measure via TRIAGE Cardiac the biological parameters of an AMI (myoglobin, troponin Ic and CKMB) and so confirm or exclude the diagnosis in certain cases. Other markers became measurable, such as BNP (brain natriuretic protein) a marker for early detection of heart failure. This natriuretic peptide is used during hospitalisation as a prognostic value in acute coronary syndrome with no cardiac insufficiency associated. More recently a semi quantitative test CardioDetect using the early release of h-FABP (heart fatty acid binding) showed a better sensibility in the first hours after chest-pain onset in out-door patients. The experience of the use of these biological bed side tests in the prehospital phase is only recent, but already permits a better management of out door patients. The future of there employ is promising. The combined use of these different markers in out door patients will probably allow in the near future identifying high risk patients. PMID- 16379108 TI - [Antithrombotics in pre-hospital phase of acute coronary syndromes]. AB - Antithrombotic therapies are the corner stone of acute coronary syndrome management. We have the proof that many of them should be initiated during the prehospital care because their clinical benefit is time-dependent. The hypothesis that anticoagulation therapy is an effective treatment of STEMI, which benefit is time-dependent, is now validated. It is also fair to affirm that GP lIb/IIIa receptor inhibitors are the adjuvant therapy of choice for primary PCI. Indeed, these medications reduce short-term and long-term mortality. This clinical benefit is time dependent. Clopidogrel therapy is probably also a medication of the prehospital phase. It is well established now that the biological efficacy of this pro drug is loading dose dependent. It is also demonstrated that its clinical efficacy depends on the time delay between symptom onset and initiation of the therapy. However, the clinical benefit of prehospital administration remains to be established. PMID- 16379109 TI - [Management of acute coronary syndromes without ST elevation: the TOSCANE registry]. AB - The problem of pre-hospital management of acute coronary syndromes without ST elevation has not been extensively studied. The practitioner is faced with three simultaneous problems: suspecting the diagnosis, how to confirm the diagnosis to introduce appropriate, rapid and effective treatment, and which prognostic criteria to use to install aggressive therapy in high risk groups (anti GP lIb/IIIa, clopidogrel, angioplasty). TOSCANE is the first multicentre French registry which analyses the impact of the emergency ambulance serve in the management of these patients. There are two objectives: to gather epidemiological data about pre-hospital and hospital management by the emergency physician and the cardiologist, and to identify at an early stage criteria of "high risk" (HR) which, according to the recommendations of the European Society of Cardiology, justify using the most aggressive therapies. From April to September 2003, 797 patients with suspected acute coronary syndromes were enrolled by 36 French centre. Of these patients, 780 were managed successfully by the emergency ambulance service and hospital cardiological department with or without a "Cath Lab", and included for analysis. The diagnosis of acute coronary syndrome without ST elevation was rarely certain in the pre-hospital period. The lack of formal paraclinical features confirming the diagnosis was often a handicap for the emergency physician. Although the European recommendations are well observed in the cardiology departments, their application and adaptability should be improved in the pre-hospital period. TOSCANE showed that all invasive strategies preceded by platelet anti-aggregant therapy in the prehospital period administered to high risk patients, significantly reduced the mortality and morbidity at one month. PMID- 16379110 TI - [Risk stratification in pre-hospital management of myocardial infarction with ST elevation: value of a risk score profile]. AB - The aims of this study were to evaluate new tools of risk stratification in an unselected population of myocardial infarction (MI), usable in a pre-hospital situation, and to compare the risk profile of these patients with those of other clinical trials or myocardial infarction registries. The risk scores of death at 30 days (TIMI score and TIMI risk index) based on data available in the context of coronary emergencies, were applied to the population base of the MI observatory of myocardial infarction in the Cote d'Or (RICO). The risk profile was expressed by the smoothed graph of frequency distribution of each score. The TIMI score applied to the RICO population had a high discriminating power (c = 0.80) for mortality whereas TIMI risk index was less powerful (c = 0.57). The risk profile of the RICO population was comparable to that of InTIME II, ASSENT 2 and the NRMI with reperfusion registry. The NRMI without reperfusion and the MAGIC studies had different profiles characterised by a shift in the graph towards high risk patients. The authors conclude that risk stratification scores, like the TIMI score, are valuable tools for early triage in the management of MI patients. The risk profiles allow comparative analysis of risk levels of populations notably with respect to other registries and also with respect to randomised clinical trials. PMID- 16379111 TI - [Pre-hospital management of acute coronary syndromes with ST elevation in the Ile de-France Region: the E-MUST registry]. AB - The E-MUST registry gathers patient data from the emergency ambulance service of the IIe-de-France for acute coronary syndromes with ST elevation seen within 24 hours from onset of symptoms. The parameters include the type of emergency phone call, details relative to the different phase of management, decisions of therapeutic strategy concerning pre-hospital thrombolysis or primary angioplasty and the different factors influencing these decisions. From January 2001 to June 2002, the mean delay from the onset of symptoms and the call-out of the emergency ambulance was 67.5 minutes for the 2584 patients studied. In this group, a pre hospital decision for coronary revascularisation was taken in 84.3% of cases, pre hospital thrombolyis started 33 minutes after arrival of the ambulance (32.7% of cases) and primary angioplasty carried out 81 minutes after that arrival (51.6% of cases). Decisions for revascularisation were less common in the elderly and those seen over 6 hours after the onset of symptoms. Pre-hospital management allows decisions concerning coronary reperfusion to be taken more often and earlier in patients with acute coronary syndromes. PMID- 16379112 TI - [Evaluation of therapeutic strategies for myocardial infarction: the ESTIM Midi Pyrenees survey]. AB - The aim of the ESTIM Midi-Pyrenees survey was to monitor the management of acute coronary syndrome with ST segment elevation by cardiologists and emergency departments in the Midi-Pyrenees region. Over a period of 2 years between June 2001 and June 2003, 1287 patients presenting with acute coronary syndrome within the first 24 hours were recruited prospectively. The initial management of these patients was undertaken either by a mobile medical team in the pre-hospital phase, or in a hospital emergency department, non-interventional cardiology department or an interventional cardiology department in 51.8%, 28.8%, 9.6% et 9.9% of cases respectively. Depending on these four modes of initial management, the median time for initial management was 1h30, 2h45, 4h30 et 4h respectively. Emergency coronary reperfusion was proposed in 89.6% of cases. Of the patients in whom reperfusion was attempted within the first 12 hours, 33.7% underwent pre hospital thrombolysis (median delay of 1h48), 35.8% underwent thrombolysis in hospital (median delay 3h), and 30.4% underwent primary angioplasty (median delay 4h40). Thrombolysis was followed by angioplasty in 80% of cases. A combined approach with thrombolysis and angioplasty was applied in 41% of patients. At one month the rate of major cardiac events, death, and/or subsequent myocardial infarction was 12%. Multivariate analysis revealed that the only significant adverse prognostic features were: not offering reperfusion [Odds ratio (OR) 4, confidence interval (CI) 2.3-3.7] and age [OR 3.8, CI 2.3-6.2]. The method of reperfusion did not influence the subsequent outcome in this regional survey. CONCLUSION: pre-hospital management allows early revascularisation. In our region there was no significant prognostic difference between pre-hospital thrombolysis and primary angioplasty. It shows that the logistic and therapeutic potentials of prehospital care are not being sufficiently exploited. PMID- 16379113 TI - [Management of patients admitted for acute myocardial infarction in France from 1995 to 2000: time to admission dependent improvement in outcome]. AB - The in-hospital management and short- and long-term outcomes was assessed in 2 registries of consecutive patients admitted for acute myocardial infarction, 5 years apart, in France. The 2000 cohort was younger and with a less frequent history of cardiac diseases, but was more often diabetic and with anterior infarcts. Time to admission was actually longer in 2000 than in 1995 (median 5.25 hours vs 4.00 hours). Overall, reperfusion therapy was used in 43% of the patients in both registries. However, the use of reperfusion therapy increased from 1995 to 2000 in patients admitted within 6 hours of symptom onset (64 vs 58%), with an increasing use of primary angioplasty (from 12 to 30%). Five-day mortality significantly improved from 7.7 to 6.1% (p < 0.03) and one-year survival was also less in the most recent period (85 vs 81%, p < 0.01). Multivariate analyses showed that the period of inclusion (2000 vs 1995) was an independent predictor of both short- and long-term mortality in patients admitted within 6 hours of symptom onset. Thus, in the real world setting, a continued decline in one-year mortality was observed in patients admitted to intensive care units for recent acute myocardial infarction, especially for patients admitted early. This goes along with a shift in reperfusion therapy towards a broader use of primary angioplasty, and with an increased use of the early prescription of recognised secondary prevention medications. PMID- 16379114 TI - [The "forgotten ones of reperfusion" in acute myocardial infarction: lessons from the ESTIM Limousin registry]. AB - The management of acute coronary syndrome (ACS) with ST elevation in daily practice does not always comply with the official guidelines. In effect, many patients do not benefit from coronary recanalisation despite being eligible. They could be described as the 'reperfusion forgotten ones'. The Limousin ESTIM study allowed us to evaluate their numbers and characteristics between 2001 and 2003. Between 1 June 2001 and 31 December 2003, 958 patients with ST+ ACS were managed within 24 hours. Among this cohort, 47% of patients did not benefit from reperfusion treatment with fibrinolysis or angioplasty. In spite of early management, the rate of non-reperfusion was significant: 30% before the third hour, and 50% between 3 and 6 hours. With univariate and multivariate analysis, the predictive features for non-reperfusion were age, length of time between onset of pain and presentation, type of admission route, absence of a call to the emergency ambulance service, and the characteristics of the ECG tracing. These data have prompted education and training, adapted for specific regions. Despite some significant improvements, the rate of non-perfusion in 2004 still remains 35% in the first 24 hours, comparable with figures in the recent literature. Being aware of this problem, taking specific action and continued evaluation with surveys like this remain important. PMID- 16379115 TI - [Primary angioplasty in acute coronary syndromes with ST-segment elevation: experience of three Alsacian centers]. AB - The guidelines of the European Society of Cardiology, published in 2003, consider primary angioplasty as the preferred treatment strategy in acute coronary syndromes with ST-segment elevation, if the procedure can be performed within 90 min after first medical contact. We report the experience of three Alsacian centers running a common prospective registry with 2504 consecutive patients enroled between January 1999 and December 2004. The average age of the patients was 62 years with a proportion of 24% women. The time delay "pain to admission" was > or =3 hours in 55.9% of the cases. The treatment delay "door to catheterisation needle" was 59 min and the mean delay "door-to-reperfusion" was 79 min. The study population was representative of the real world including subsets of patients with a particulary high risk profile: age > or =70 years in 33%, a Killip grade > or =3 in 11.5%, rescucitated sudden death in 6.6% and cardiogenic shock in 10.9% of the patients respectively. The immediate procedural success rate (Timi 3 flow) in the treated coronary artery was 96.5%. The overall inhospital mortality-rate was 9.3%. The combinations of rescucitated sudden death -cardiogenic shock or age > or =75 years--cardiogenic shock were associated with a poor clinical outcome and mortality rates of 69% and 72.6% respectively, where as in the absence of abovementioned clinical high risk settings, the mortality rate was as low as 1.4%. The overall bleeding complication rate was 1.4%. The policy of systematic primary angioplasty in acute coronary syndromes with ST Segment elevation appears to be coherent. The procedural complications and the in hospital mortality rates were low, except in the presence of above mentioned clinical high risk settings. PMID- 16379116 TI - [Thrombolysis: Bucarest registry]. AB - The end-point of this retrospective study was to evaluate the standard of care in terms of in-hospital morbidity and mortality for patients with acute myocardial infarction (AMI) treated by thrombolysis within the first 12 hours from the start of the symptoms in five cardiology centers from Bucarest for a period of 5 years (2000-2004). This retrospective registry on a central database included 1814 patients (73.63% men, mean age 59.9 +/- 11.8 years), presented in an average time of 211.63 minutes from pain start. The most frequently used fibrinolitic was streptokinase (66.21%), administered most often in 30 minutes and for a subgroup in 20 minutes--accelerated regimen, with a good efficiency for the reperfusion of the culprit vessel evaluated non-invasively (clinical, electrical and biological methods). The global in-hospital mortality was 11.1%. The only predictors of in hospital mortality were female gender and advanced age (>75 years) [p < 0.05]. The rate of haemorrhagic complications was not different from the one described by other clinical studies. The treatment by anticoagulants, antiaplatelets agents like aspirin, ACE inhibitors and statins were significant determinants of in hospital survival. In the subgroup followed-up for 1 year (315 patients), the most frequent complication was the heart failure. In conclusion, in Bucarest, where availability of primary angioplasty in AMI was limited, thrombolysis with streptokinase was still very much used, with acceptable low in-hospital mortality and relatively high rate of artery reperfusion appreciated by non-invasive methods. PMID- 16379117 TI - Mercury emissions: is there a global problem? PMID- 16379118 TI - A critical review of economic valuation studies of externalities from incineration and landfilling. AB - The primary objective of this study was to assist waste management researchers, decision-makers and waste managers at national, regional and local levels, in their decision-making processes, with most recent valuations on the environmental and social costs of externalities associated with various pollutants and disamenities related to landfilling and incineration of municipal solid waste. The aim was achieved by mapping, gathering, analysing, comparing and synthesizing various valuation estimates, based on a thorough review of existing literature. This study provides the first comprehensive review and analysis focused on primary and secondary valuation studies, conducted since 1990. The second objective was to assess the appropriateness and reliability of the valuation methods and techniques that were performed in the reviewed studies. The results of the review are summarized in tables, organized by topics and units of measure and in addition a classified list that describes the profile of the reviewed studies is provided. The results are then analysed and compared, and recommended ranges of the values are presented. The study reveals inconsistency in part of the estimates across the reviewed studies and provides reasonable explanations for the variations. Given the nature of uncertainty, and the difficulties associated with transferring values among different places and cases, these values should be considered mostly as an indication for the order of magnitude of the externalities. Nevertheless, these essential estimates of the external costs can beneficially be used with proper adjustment for each individual case to address important policy questions regarding landfilling and incineration of waste. PMID- 16379119 TI - Transboundary hazardous waste management. Part I: Waste management policy of importing countries. AB - Mixed metal-containing waste, polychlorinated biphenyls (PCB) containing capacitors, printed circuit boards, steel mill dust and metal sludge were among the most common wastes exported from Taiwan. Before the implementation of the self-monitoring model programme of the Basel Convention (secretariat of the Basel Convention 2001) in the Asia region, Taiwan conducted a comprehensive 4-year follow-up project involving government authorities and the waste disposal facilities of the importing countries. A total of five countries and nine plants were visited in 2001-2002. The following outcomes can be drawn from these investigations. The Chinese government adopts the strategies of 'on-site processing' and 'relative centralization' on the waste management by tightening permitting and increasing site inspection. A three-level reviewing system is adopted for the import application. The United States have not signed the Basel Convention yet; the procedures of hazardous waste import rely on bilateral agreements. Importers are not required to provide official notification from the waste exporting countries. The operation, administration, monitoring and licensing of waste treatment plants are governed by the state environmental bureau. Finland, France and Belgium are members of the European Union. The procedures and policies of waste import are similar. All of the documents associated with transboundary movement require the approval of each government involved. Practically, the notification forms and tracking forms effectively manage the waste movement. PMID- 16379120 TI - Comparison of external costs between dry tomb and bioreactor landfills: taking intergenerational effects seriously. AB - Dry tomb and bioreactor landfills were analyzed with respect to their external costs in an intergenerational cost-benefit analysis in a partial framework which enabled a sounder comparison to be carried out between these two technologies from a socio-economic viewpoint. Obviously, this approach was only a first step for building a comprehensive basis of any environmental as well as fiscal policy in the field of waste management. All external costs are identified and evaluated in three different scenarios, corresponding to a worst case, a best guess and a best case. Obviously, discounting is crucial with respect to an intergenerational perspective. Generation-adjusted discounting (GAD) was applied to take into account equity as well as efficiency criteria, in order to deal with three different types of uncertainties that are decisive in waste policy decisions: a physical uncertainty is captured by introducing our three different scenarios; a macroeconomic uncertainty is taken into consideration by calculating present values using different real growth rates; and a microeconomic uncertainty is taken into account by considering individual peculiarities reflected in their subjective time preference rate. The findings show, that whenever there is a low real GDP growth of less than 1%, the bioreactor is generally superior to the dry tomb (lower present values of external costs). This statement becomes more valid as the growth rate decreases. However, whenever there are high positive growth rates, it is the dry tomb technology which is superior to the bioreactor system. PMID- 16379121 TI - Quality control of waste to incineration--waste composition analysis in Lidkoping, Sweden. AB - In order to decrease environmental impacts in waste management the choice of treatment method must be based on the characteristics of the waste. Present sampling procedures do not provide statistically representative samples of solid waste and this provides difficulties in characterization. The objective of this study was to develop a procedure for waste component analysis and sampling of waste after collection and at plant level. A further objective was to characterize the waste delivered to an incineration plant for physical and chemical properties and to determine the amounts of delivered waste that could be classified as biofuels and fossil fuels. The proportions of recyclables and hazardous waste were also examined. Samples were taken randomly from waste trucks and divided by square implementation. Statistical analysis of the results showed that the number of sub-samples could be decreased with only a moderate increase in the confidence interval. This means that future waste composition analyses could be made more efficient and thereby less expensive. The analysis of the waste delivered to the Lidkoping incineration plant (Central Sweden) showed that 66.4% of the household waste was composed of biofuels and 21.3% of non-renewable combustibles, of which 40.3% were recyclables. In addition, 11.6% of the household waste was non-combustible and 0.6% hazardous waste. The heat value for the biofuels was 18.0-19.7 MJ kg(-1) dry mass (DM) and for the fossil fuels 28.2 33.9 MJ kg(-1) DM. The industrial waste consisted of 35.9% biofuels, 62.0% fossil fuels, 1.6% non-combustible and 0.5% hazardous waste. The heat value was 19.5 MJ kg(-1) DM for the biofuels and 31.4 MJ kg(-1) DM for the fossil fuels. PMID- 16379122 TI - Separation of fine binary mixtures under vibration in a gas-solid fluidized bed with dense medium. AB - The coke-slag mixture discharged from water-air generators not only causes a serious environmental problem, but also is a potential resource. In order to obtain a high economic return, separation of fine binary mixtures in a vibrating fluidized bed with dense media was experimentally investigated. The effects of volume ratio (mono-component volume to dense medium volume or binary mixture volume to dense medium volume), the fluidization velocity and vibration parameters on separation efficiency are discussed. The experimental results show that vibration can prevent the coalescence and growth of bubbles and improve fluidization properties of particles and the stability of bed density. The major factors associated with separating efficiency are discussed. Based on the experiments, the ratio of the volume of particles to bed volume and the vibration conditions are also reported. It was found that the separation efficiency for a fine coke-slag mixture was greater than 70% and close to 80% under optimal conditions. PMID- 16379123 TI - Remediation of an oily leachate pond in Estonia. AB - Until recent years, waste oil and oil-contaminated waters commonly ended up in landfills. At some dump sites, ponds of oily liquids and leachate were formed. To remediate such ponds, an interdisciplinary approach is now required, keeping costs at an affordable level, particularly in countries with changing economies. From 1974 to 1993, liquid oily wastes taken to the Laguja landfill, in Estonia, were disposed of in a pond with a surface area of 9800 m2. It was estimated that the pond contained 4500-6000 m3 of oily water and 3500 m3 of oil-containing bottom sediments. This study aimed at developing an environmentally sound and cost-effective method for remediation of the oily liquids, leachate and contaminated underlying sediment material, to meet the existing legal demands. It was concluded that treatment of contaminated water is well established and the procedures carried out to meet the regulatory demands achieved satisfactory results. However, regarding treatment of sediments it was concluded that legal and technological aspects, as well as monitoring procedures are not fully established and are usually underestimated. Laboratory investigations can provide valuable information in decision-making, and contribute to effective full-scale remediation planning. PMID- 16379124 TI - Physico-chemical treatment of marble processing wastewater and the recycling of its sludge. AB - In the first part of this study, the treatability of marble processing wastewater by the coagulation-flocculation process was investigated. Optimum coagulant flocculant doses for turbidity removal in wastewater from the cutting, faience and equalization processes were determined as 500, 200 and 500 ppm of Al2(SO4)3; 300, 500 and 300 ppm of FeCl3 and 600, 400 and 200 ppm of Agrofloc 100 (AGRON Water Treatment Technologies and Chemical Marketing Industry and Trade Limited Company, Izmir, Turkey), respectively. It was found that the removal of total solids from cutting and equalization process wastewaters was highest for the 100 ppm dosage of all chemicals used. The amount of total solids removed from faience process wastewater by Agrofloc 100 was higher than that removed by the other chemicals used. The removals of suspended solids from cutting, faience and equalization process wastewaters were similar to each other for each of the chemicals. The pH values after treatment by Agrofloc 100 were higher than the values determined after treatment by other chemicals for all process wastewater. Electrical conductivity values, however, were lower for Agrofloc 100 than for the others. Settled sludge volume experiments showed that settled sludge volumes decreased with time. The results of the quiescent settling experiment showed that the settling type could be termed flocculent settling. In the second part of the study, the usage of waste sludge from marble processing as an additive material in cement was investigated. The waste sludge originated from the wastewaters of different steps of the marble processing plant. Waste sludge was replaced with cement at various percentages by weight to prepare the mixtures of mortar. The specimens poured into the moulds were held for 24 h, removed from the moulds and held again for 28 days in lime-saturated water at 23 degrees C. Compressive and flexural strengths were evaluated with respect to percentages of waste sludge replaced with cement. The maximum compressive and flexural strengths were observed for specimens containing a 6% waste sludge when compared with control and it was also found that waste sludge up to 9% could effectively be used as an additive material in cement. PMID- 16379125 TI - Physico-chemical treatment of Merida landfill leachate for chemical oxygen demand reduction by coagulation. AB - In order to determine the optimal dosage and type of coagulant for the physico chemical treatment of leachate from the sanitary landfill of Merida, Mexico, a total of 864 jar tests were performed. Four metallic coagulants (ferric chloride, ferric sulphate, aluminium polychloride and aluminium sulphate) with doses ranging between 50 and 300 mg L(-1) and two polyelectrolytes (high-density anionic and cationic reagents) with doses from 2 to 12 mg L(-1) were tested. Neither an adequate type of coagulant nor an optimal dose could be found. The removal of contaminants was measured as total and dissolved chemical oxygen demand (COD). Soluble COD removal efficiencies were low, from 0 to 47%, with a 4% average value only. These low values of organic material removal were attributed to the particular characteristics of the Merida landfill leachate (low suspended solids concentration), so even with sweep-floc coagulation (300 mg L(-1) dose) only low COD removal efficiencies were obtained. A study of the suspended particle size distribution of the leachate was conducted in order to explain the poor performance. The particle size distribution ranged from 0.375 to 948.2 microm, with an average value of 22.97 microm. In a second step the optimal pH for physico-chemical treatment of these leachates was determined. Finally a greater than 90% removal of organic material, measured as suspended COD, was obtained at pH 2, which was considered as the optimal value. PMID- 16379126 TI - Solid waste collection and recycling in Nibong Tebal, Penang, Malaysia: a case study. AB - The implementation of a suitable solid waste management programme with appropriate methods of recycling as an inherent element is vital to the alleviation of the problems associated with solid waste generation, handling and disposal, environmental conservation, public hygiene, etc. The present work is a case study on solid waste collection and recycling practices in Nibong Tebal town, Penang, Malaysia. The amount and types of domestic waste generated, household participation in recycling, identification of existing problems related to the implementation of the recycling programme, etc. formed the basis of this study. Surveys (interviews/questionnaires) and on-site observations were conducted to gather information on the solid waste collection and recycling practice of the residents. A focus group of 60 individuals was selected and their response to a questionnaire, prepared according to a Likert scale, was obtained and analysed. The majority of the respondents expressed concerns about recycling and wanted more to be done in this regard. Illegal collection, aesthetically displeasing sites and a lack of public awareness were problems of major concern. Issues related to inadequate funding and manpower as well as end market are also addressed and suggestions made. PMID- 16379127 TI - Health-care waste management in Lao PDR: a case study. AB - This study investigated the health-care waste (HCW) management at each health care facility level at two selected sites in the Lao People's Democratic Republic (Lao PDR): Vientiane Municipality; and Bolikhamxay province. It focused on the amount of HCW, its segregation and the factors influencing HCW management, particularly segregation procedures. A high proportion of incorrectly segregated medical waste was found at each level of health-care facility. Re-segregation revealed 39, 62, 57 and 37% at national hospital, provincial hospital, district hospital and health centre level, respectively, was poorly segregated. The mean of generated HCW was 0.62 kg/bed per day (Vientiane Municipality) and 0.38 kg/bed per day (Bolikhamxay) at two study sites. A higher proportion of medical waste (MW) from the inpatient department at the primary health-care level was found. Thus, HCW management at primary health-care facilities needs more attention and should be better understood. PMID- 16379128 TI - What are neuroscience nursing best practices? PMID- 16379129 TI - Successful outcome in severe traumatic brain injury: a case study. AB - This case study describes the management of a 54-year-old male who presented to the Hospital of the University of Pennsylvania (HUP) with a traumatic brain injury (TBI) after being assaulted. He underwent an emergent bifrontal decompressive hemicraniectomy for multiple, severe frontal contusions. His postoperative course included monitoring of intracranial pressure, cerebral perfusion pressure, partial pressure of brain oxygen, brain temperature, and medical management based on HUP's established TBI algorithm. This case study explores the potential benefit of combining multimodality monitoring and TBI guidelines in the management of severe TBI. PMID- 16379130 TI - An update on transient ischemic attacks. AB - Each year in the United States 200,000-500,000 people have a transient ischemic attack (TIA). These episodes of brief neurologic deficits were thought to be fairly benign, but this view is changing. In 2002, a new definition for TIA was proposed, and a more intensive diagnostic workup recommended to look for a probable cause of the transient neurologic deficit. Implementation of prevention strategies is essential because the risk of a stroke following a TIA is approximately 30% within 5 years. These changes have important implications for nursing care and patient advocacy. In addition, patient and family education must be addressed by the entire healthcare team. PMID- 16379131 TI - Effects of a cognitive-behavioral program for women with multiple sclerosis. AB - The purpose of this quasi-experimental study was to evaluate the effectiveness of a cognitive-behavioral intervention for women with multiple sclerosis (MS). Thirty-seven adult women with MS participated in a group-based intervention program titled "Beyond MS," which was led by master's-prepared psychiatric nurses. For participants, the program involved reading a manual and meeting for five weekly group sessions. Perceived health competence, coping behaviors, psychological well-being, quality of life, and fatigue were measured at four time periods: 5 weeks before the beginning of the intervention, immediately before the intervention, at the end of the 5-week intervention, and at a 6-month follow-up. There were significant improvements in the participants' perceived health competence (p < .01), indices of adaptive and maladaptive coping (p < .04), and most measures of psychological well-being (p < .05) from pre- to postintervention. The positive changes brought about by this relatively brief intervention program were maintained during the 6-month follow-up period. This cognitive-behavioral intervention has also been used effectively in the rheumatoid arthritis population and may be adaptable to benefit individuals with other chronic conditions. PMID- 16379132 TI - Cerebral venous sinus thrombosis. AB - Cerebral venous sinus thrombosis (CVST) is a rare and potentially deadly condition. Common etiologies include hypercoagulable diseases, low flow states, dehydration, adjacent infectious processes, oral contraceptives, hormonal replacement therapy, pregnancy, and puerperium. Symptoms include nausea, seizures, severe focal neurological deficits, coma, and headache (the most common presenting symptom). Anticoagulation is the mainstay of treatment for CVST. Transvenous clot lysis can be performed using injected thrombolytic agents and specialized catheters for clot retrieval. PMID- 16379133 TI - Mothering a 3- to 6-year-old child with hemiparesis. AB - Stroke is estimated to occur in approximately 2.5-3.2 children per 100,000 annually. While some aspects of mothering a disabled child are currently known, there are other aspects of this experience that remain unknown. Much of the research focuses on children who are severely disabled, thus leaving a gap in what is known about the experience of mothers of mildly to moderately disabled children. Little is understood about mothering children who have a less severe disability, and in particular, a 3- to 6-year old child with hemiparesis. Therefore, the purpose of this study was to describe the experience of mothering a 3- to 6-year old child with hemiparesis. This study incorporated a phenomenological approach using Colaizzi's descriptive methodology. The participants were five biological mothers of children aged 3-6 years who were diagnosed with hemiparesis following a stroke. The children were considered mildly to moderately disabled and required two to three therapies, such as physical therapy, occupational therapy, speech therapy, or special education. In depth, face-to-face interviews were conducted, audiotaped, and transcribed verbatim. By using Colaizzi's method for data analysis, the following six themes emerged: (1) Mothers' Way to Know, (2) Being Blown Away, (3) Gradual Awakening to an Uncertain Future, (4) Fiercely Advocating for My Child, (5) Living in a Middle World, and (6) A Heart Filled with Joy and Sorrow. This study provided data that were used to provide a beginning description of the experience of mothering a child with hemiparesis. PMID- 16379134 TI - Tension pneumocephalus. AB - Pneumocephalus, air in the cranial cavity, is a common occurrence following cranial surgery. However, tension pneumocephalus, accumulation of air trapped in the cranial cavity causing a mass effect and abnormal neurological signs, is uncommon. Knowledge about conditions that contribute to tension pneumocephalus, a potentially fatal complication, including the related signs and symptoms, is imperative. Immediate collaboration with a physician allows for timely treatment and patient recovery. Treatment measures include preoperative teaching, immediate removal of intracranial air, supine positioning, administration of 100% oxygen, repair of the bony and dural defect, and, if indicated drain placement into the air cavity, temporary tracheotomy, and antibiotics. PMID- 16379135 TI - Brain tissue oxygen practice guidelines using the LICOX CMP monitoring system. AB - When a new technology is introduced it is important to empower the bedside practitioner with a resource tool that outlines the purpose, placement procedure, technology application guidelines, and interventions associated with that new technology. This promotes product and patient safety and successful implementation of the new technology. Continued evaluation of bedside clinical practice and the technology used in the care and treatment of the severe brain injured patient can lead to improvements in management and in technology design. Future clinical research initiatives exploring the impact of new technology will enable us to discover cost-effective treatments and interventions that will improve the outcome for a person with traumatic brain injury, a condition that devastates hundreds of thousands of Americans each year. PMID- 16379136 TI - Nurses making a difference: the process of technology transfer. PMID- 16379137 TI - Salt fluoridation--an alternative in automatic prevention of dental caries. AB - Despite great improvements in terms of reduced prevalence and amount of dental caries in populations worldwide, problems still persist particularly among the underprivileged groups of both developed and developing countries. Research and practical experience gained in several countries have demonstrated however, that dental caries can be prevented effectively through establishment of fluoride programmes. Water fluoridation, salt fluoridation, milk fluoridation and use of affordable fluoridated toothpastes play the major roles in public health. The present paper outlines the relevance and some practical aspects in relation to implementation of salt fluoridation programmes. The World Health Organisation Oral Health Programme provides technical assistance to countries in the process of planning, implementing and evaluating salt fluoridation projects. PMID- 16379138 TI - Dental caries, its surface susceptibility and dental fluorosis in South India. AB - OBJECTIVES: To correlate water fluoride levels with dental caries and dental fluorosis in areas with different levels of fluoride in the drinking water and to establish the surface susceptibility of dental caries in an endemic fluoride area. METHODS: 544 schoolchildren 12 to 15 years of age from the Davangere region of India were examined. The DMFS index was used to measure dental caries, which was further differentiated into smooth surface and pit and fissure lesions. Dean's index was used to diagnose dental fluorosis. Five villages with fluoride levels ranging from 0.43 ppm to 3.41 ppm were studied. RESULTS: There was a highly significant negative correlation (r = -0.16) between water fluoride levels and dental caries. Dental fluorosis increased from 16% at 0.43 ppm to 100% at 3.41 ppm. Pit and fissure lesions made up the vast majority of the lesions in all the villages and showed a decreasing trend with increasing fluoride levels, however no such trend was seen for smooth surface lesions. CONCLUSIONS: Water fluoride was an important factor responsible for the low caries prevalence. The prevalence of fluorosis and low caries even in low fluoride areas may point to a halo effect. PMID- 16379139 TI - Cocaine abuse: orofacial manifestations and implications for dental treatment. AB - Millions of individuals in Europe and the United States use cocaine regularly. Use of cocaine may have several orofacial effects such as perforation of nasal septum and palate, gingival lesions and erosion of tooth surfaces. Recent use of cocaine may also increase medical risks during dental treatment, especially when local anaesthetics with epinephrine or epinephrine-impregnated retraction cords are used. Therefore, it is recommended to postpone dental treatment at least 6 to 24 hours after the use of cocaine. PMID- 16379140 TI - Estimation of global DMFT for 12-year-olds in 2004. PMID- 16379141 TI - Ranking countries by dental status using the DMFT and FS-T indices. AB - AIM: To test the hypothesis that the DMFT index does not adequately reflect the dental status as well as the FS-T index by comparing the country rankings of the DMFT and the FS-T indices in 18 and 35-44 year-olds in a range of countries. The FS-T is the number of filled or sound teeth and is a measure of functional status. METHOD: The DMFT data from WHO for 18-year-olds and 35-44-year-olds was used and FS-T index values were estimated for the 18-year-olds for 12 countries and for 35-44 age groups for 30 countries. RESULTS: The ranking by DMFT index scores was lower for developed countries than for developing countries. Despite the relatively high DMFT index in developed countries, their FS-T index was higher and therefore the developed countries ranked higher by FS-T compared to their ranking by DMFT. CONCLUSIONS: These findings suggest that it may be more useful to use the FS-T index with the DMFT index in studies comparing dental status between countries. PMID- 16379142 TI - Partnering globally and acting locally for infection control in Nepal. AB - AIMS: To describe the process and outputs of an international collaborative oral health promotion project to develop a national infection control policy and training programme for oral health care workers in the low income country of Nepal between April, 2003 and May, 2004. METHOD: The project process was implemented in phases: 1) extensive review of national and international infection control documents; 2) development of draft infection control policies and protocols; 3) development of instruments to assess knowledge, attitude and behaviour and infection control practices; 4) baseline survey to assess these factors; 5) development of training programmes and training of oral health care providers; 6) revision of infection control policies and protocols, survey questions, assessment instruments, and training programme. OUTPUTS: Project outputs include a national infection control policy and protocols, infection control assessment instruments, infection control training materials and programmes, and oral health care providers trained in infection control. SUMMARY: The results of the project to develop a national infection control policy and training programme for oral health care workers in dental clinics, dental education institutions and the Nepal Primary Health Care System, required the collaboration of policy makers, health professionals, health managers, oral health care providers and educators from the government sector, private sector, Non-Government Organisations (NGOs) and International Non-Government Organisations (INGOs). PMID- 16379143 TI - Public perception of cross-infection control in dentistry in Nigeria. AB - OBJECTIVES: To assess the perception of patients on the use of gloves and facemasks in dental practice, methods of sterilisation and the risk of cross infection. METHODS: Questionnaire survey of patients attending two teaching hospitals and a private dental clinic in Lagos, Nigeria. RESULTS: 228 respondents returned the questionnaires. Nearly all (98.6%) regarded the use of gloves as necessary and 189 (82.9%) did not mind the use of gloves, only 129 (56.6%) felt the facemask was necessary although 132 (57.9%) did not mind its use. More than half of the respondents (54%) felt they could contract an infection in the dental clinic and 43% of them identified HIV as a risk. This response was significantly affected by educational status (p<0.05). 140 patients (61.4%) would not attend a clinic if they knew that HIV patients were treated there. Females were significantly more unwilling to attend (p<0.05). CONCLUSION: While this survey shows a high level of acceptance of the use of gloves, there is a glaring lack of knowledge and acceptance of the use of the facemask indicating a need for more effective public enlightenment on the use of barrier methods. It also reveals a great deal of fear for HIV among Nigerians. PMID- 16379144 TI - 21st century endodontics. Part 5. AB - This series of papers has attempted to present the latest principles in root canal treatment. It is hoped that by comparing the techniques described with their own clinical procedures, the reader will have considered whether changes may be appropriate. These may have been immediate, for example introducing an EDTA solution as an additional irrigant, or more long-term, perhaps by planning to attend hands-on courses to perfect a new procedure before using it in vivo. Professor Dick Walker, an excellent and eminent endodontist, once said that he would not even think of doing something to a patient that he had not done at least twenty times, (and probably more), on an extracted tooth. Wise words indeed! Unfortunately even with practice and persistence, problems may arise during or after root canal treatment. The focus of this paper is on some of the more common problems that may be encountered and how endodontic treatment failures may be addressed. PMID- 16379145 TI - Dentist-patient relationship and quality care 5. Modification of behaviour. AB - High quality clinical outcome, good oral health and prevention of oral diseases depend heavily on the patient's adherence to suggested behaviour. Improved awareness of psycho-social determinants of patient's health behaviours may aid to modify patient behaviour, increase patients' adherence, contribute to general health and quality of life of individual patients and the public and maximise the benefits that dentistry can offer. PMID- 16379146 TI - Avian influenza, China--update. PMID- 16379147 TI - Index, Volume 80, 2005, Nos. 1-52 Subject index. PMID- 16379148 TI - A most marvelous time. PMID- 16379149 TI - The greatest pleasure of leadership is meeting and working with so many concerned dentists who are willing to donate their valuable time to help solve the problems that develop in our profession. PMID- 16379150 TI - "The truth about children's identification products," June 2005. PMID- 16379152 TI - Clinical operating microscopes: they're not just for endodontists anymore. PMID- 16379153 TI - Dental health professionals vital in reducing effects of influenza. PMID- 16379155 TI - [The basic and clinical studies of moxifloxacin: the profile as respiratory quinolones]. PMID- 16379156 TI - [In vitro activity of antimicrobial agents against clinical isolates of Pseudomonas aeruginosa]. AB - Two hundred and seven clinical isolates of Pseudomonas aeruginosa were collected at Tenri Hospital between April 2003 and March 2004. We determined the minimum inhibitory concentration (MIC) of 16 antimicrobial agents, including prulifloxacin, pazufloxacin and biapenem which were recently published in Japan, against these isolates according to the guidelines of the Clinical and Laboratory Standards Institute. For the fluoroquinolones, the rank order of activity was prulifloxacin (MIC50, 0.5 microg/ml)>ciprofloxacin (1 microg/ml)> pazufloxacin (2 microg/ml)=levofloxacin (2 microg/ml)>gatifloxacin (4 microg/ml). For the carbapenems, the rank order of activity was meropenem (MIC50, 1 microg/ml)=biapenem (1 microg/ml)>imipenem (2 microg/m)>panipenem (8 microg/ml). For the cephalosporins and monobactam, the overall rank order of activity was cefozopran (MIC50, 4 microg/ml)= ceftazidime (4 microg/ml)>cefepime (8 microg/ml)=piperacillin/tazobactam (8 microg/ml)>aztreonam (16 microg/ml)= cefoperazone/sulbactam (16 microg/ml)=cefpirome (16 microg/ml). The rates of susceptibility to antimicrobial agents as per the criteria of the Japanese Society of Antimicrobial Chemotherapy were especially high for cefozopran (63%), biapenem and meropenem (61%), and pazufloxacin (53%) and ciprofloxacin (53%). These findings suggest that prulifloxacin, pazufloxacin and biapenem, which are newly introduced, are clinically effective in the treatment of infection caused with P. aeruginosa. PMID- 16379157 TI - [A preliminary study of the administration of carbapenem antibiotics in sepsis patients on the basis of the administration time]. AB - It is important to take into consideration the duration for which the plasma concentration of the drug is higher than the Minimal Inhibitory Concentration during treatment with carbapenem antibiotics, because the antibiotics are time dependent drugs. A preliminary study of the administration of carbapenem antibiotics on the basis of the pharmacokinetics/pharmacodynamics (PK/PD) was conducted. SUBJECTS: Ten patients with intraabdominal infection. METHODS: The patients were divided into two groups: the first group was assigned to administration of a carbapenem antibiotic (meropenem) at a daily dose of 0.5 g in 3 divided doses, each dose by intravenous infusion over 3 hours (Group 3 H), and the other group was assigned to administration of each dose over 30 minutes (Group 30 M). The body temperature (BT), white blood cell count (WBC), serum C reactive protein (CRP) level, and the systemic inflammatory reactive syndrome (SIRS) score before and 96 hours after the drug administration were compared between Group 3 H and Group 30 M. RESULTS: There were 5 patients (mean age, 67.4+/-14.6 years) in Group 3H and 5 patients (mean age, 60.0+/-12.8 years) in Group 30 M. The evaluated parameters (BT, WBC, CRP, SIRS score) before the drug administration in Groups 3 H and 30M were not significant. Group 3 H showed significant decreases in the SIRS scores at 96 hours after the drug administration, however, there were no significant differences in the BT, WBC or CRP between the two groups. DISCUSSION: Group 3 H showed early improvement in the SIRS scores. Administration of carbapenem antibiotics based on the PK/PD is important, and requires further studies. PMID- 16379158 TI - [Surveillance for various injectable antimicrobial susceptibility among clinical isolates of Pseudomonas aeruginosa in hyogo prefecture]. AB - Antimicrobial susceptibility of 13 antimicrobial drugs for the injection and O group antigen serotype were measured for the 766 Pseudomonas aeruginosa strains that had been isolated from various clinical materials in 29 facilities in the Hyogo prefecture from April to September in 2004. Metallo beta-lactamase detection was also performed. The antimicrobial activity was excellent in the order of GM, MEPM, AMK, CPFX and CAZ. Susceptible category of the breakpoint by National Committee for Clinical Laboratory Standards (CLSI/NCCLS) was excellent in the order of AMK, GM, PIPC, CZOP, and MEPM. As for the susceptibility of Carbapenem, it is confirmed that susceptible of MEPM was detected in 47 strains (36.4%) and metallo beta-lactamase producing P. aeruginosa was in 3 strains (0.4%) and multi-drug resistant P. aeruginosa were in 7 strains only (0.9%) among 129 strains of the IPM resistant (I or R). The results of the susceptibility test against P. aeruginosa were different in each facility, but there were several stocks having the identical O-antigen serotype and anti-biogram pattern in some facilities. The nosocomial infection measures including the antimicrobial propriety use are necessary. PMID- 16379159 TI - [Outline of the drug use result survey of Firstcin (cefozopran hydrochloride) as part of postmarketing surveillance in Japan]. PMID- 16379160 TI - Validity of an instrument assessing oral health problems in people with Down syndrome. AB - AIM: The aim of this study was to validate a proxy measure of oral health designed to be completed by the English-speaking parents of people with Down syndrome (DS) aged four years or more. METHODOLOGY: Items were generated through literature review, interviews with parents of people with DS and professional experts and through frequency testing. Data were gathered from one population based and two clinic-based samples for the separate aspects of validation. Validation consisted of evaluation of: i) internal reliability of the domain structure through Cronbach's alpha; ii) criterion validity against clinical indicators and a clinician's evaluation of some items; iii) construct validity involving an age-matched comparison of domain scores between people with DS and non-DS siblings, and within the DS group by health status indicators; and iv) test-retest reliability through the generation of intra-class correlation coefficients (ICC). RESULTS: A 20-item instrument with four domains (communication, eating, parafunction and symptoms) was developed. Cronbach's alpha by domain was 0.5-0.8. Indicators of criterion validity for domains against clinical indicators (Spearman's coefficient 0.1-0.4) and parent-rated items against clinician-rated items (weighted Kappa 0.1-0.8) were varied as anticipated. Indicators of construct validity (differences with non-DS siblings and correlations with medical status within the DS group) were excellent. Test retest reliability was good (ICC range 0.64-0.84). CONCLUSION: These data suggest the test instrument is valid as a descriptive, discriminative, proxy English language measure of oral health problems in people with DS aged four years or more. PMID- 16379161 TI - Subjective impacts of dental caries and fluorosis in rural Ugandan children. AB - OBJECTIVE: Describe the oral health related quality of life among a group of children in rural Uganda and compare impacts on oral health related quality of life associated with dental caries and fluorosis. BASIC RESEARCH DESIGN: Cross sectional clinical and questionnaire analytical study. PARTICIPANTS: Proportional sample of 174 12 year olds attending primary schools in a rural sub-county of Uganda. OUTCOME MEASURES: Clinical assessments using WHO basic methods and the Thylstrup and Fejerskov index of Fluorosis (TFI). Child Oral Health Related Quality of Life data collected with self-administered child perception questionnaire (CPQ11-14). RESULTS: Two thirds of children reported a dental impact 'often' or 'everyday'. The mean number of impacts per child at this threshold was 2.6 and the mean total CPQ11-14 score was 25.8 (sd 21.1). Mean DMFT was 0.68. No children had fillings. Forty-one children had dental fluorosis with 10 having scores greater than 2. CPQ11-14 showed acceptable criterion validity and reliability. The number of sites with gingivitis or the presence of calculus or trauma were not associated with summary measures of CPQ11-14 whereas having any dental caries or treatment experience was associated with higher total scores and more impacts. Socially noticeable fluorosis (TFI >2) was associated with more impacts but not with higher total scores. CONCLUSIONS: Despite low levels of oral disease these children experience appreciable impacts on oral health related quality of life. The greatest burden was associated with dental caries and to a lesser extent, fluorosis. PMID- 16379162 TI - Assessing sealant retention using a Poisson frailty model. AB - OBJECTIVE: This paper utilises a Poisson frailty regression model with clustering to report on the failure rates of sealants placed and repaired during a three year follow-up school-based comprehensive preventive programme. BASIC RESEARCH DESIGN: The programme included annual application/reapplication of light-cured fissure sealants in 210 12-year-old subjects over three years. Sealants placed at the commencement of each year of the programme were assessed for retention 12 months later and repair/reapplication was undertaken at that time for those surfaces judged to require it. An application of a Poisson regression model with random effects ("frailties") and clustering was used in this paper to allow for heterogeneity between subjects. Whilst it appears that the direct utilization of frailty models in determining fissure sealant failure rates has not been reported, it is clearly an appropriate use of this statistical method. A total of 1,544 sealants were placed during the study period. Of these, 1,038 (67.2%) were first placements at baseline and 506 (32.8%) were repairs/reapplications. Of these repairs/reapplications, 206 (40.7%) were performed at first year follow-up and 300 (59.3%) at second year follow-up. RESULTS: The Failure Rate Ratios were statistically significantly greater for second molars cf. first molars (FRR=1.33, p=0.008) and for lower molars cf upper molars (FRR=1.32, p<0.001). Failure rate ratios for sealants placed in the distal occlusal pit and fissures cf. mesial occlusal pit and fissures were significantly higher (FRR=1.33, p<0.001). A significant interaction was found for molar type versus pit and fissure site (p=0.009). There was no significant interaction between molar type and arch. CONCLUSIONS: Sealants placed on second molars, those placed on lower molars and those placed on the distal fissure sites have a higher failure rate. As individual tooth sites in the mouth are not independent, any statistical analysis should allow for these associations when assessing sealant retention. PMID- 16379163 TI - Summary measures of caries prevalence to describe high-risk communities. AB - STUDY OBJECTIVE: This investigation compared the Significant Caries Index (SiC) with alternative summary measures of caries prevalence, designed to highlight high-risk communities and examined the implications of their use in health policy and planning in Wales. DESIGN: Data from the 2001 BASCD survey of 7,412, 12-year old Welsh children were used in this analysis. As an alternative to the 33% cut off value utilised in the SiC, the mean DMFT of children with the 10, 20 and 25% highest caries scores were calculated for 22 local health board areas, as was the mean DMFT for the whole population, %DMFT >0, and the mean for those with DMFT >0, >1, >2, and >3. SETTING: Examinations for the childhood epidemiology programme took place in Welsh secondary schools. PARTICIPANTS: 12-year-old children examined in the 2000/01 survey. MAIN RESULTS: The mean DMFT for the whole population was 1.31, and the mean DMFT of those with caries was 2.56. The mean DMFT for SiC 33%, 25%, 20% and 10% was: 3.39, 3.85, 4.28, and 5.31 respectively. When local health board areas were ranked according to mean DMFT score at different percentage cut-off points, variation in rank order was observed, the greatest difference in rank order being 6. Furthermore, when compared with summaries based on DMFT scores traditionally used in Wales, even greater rank differences were noted (up to 14 places). CONCLUSIONS: Rank order of geographical area is influenced by the definition of the index chosen when describing high risk populations using a variety of caries indices. This has implications for health policy planning and resource allocation. PMID- 16379164 TI - The process and outcome of a programme for preventing early childhood caries in Thailand. AB - OBJECTIVE: To evaluate the process and outcomes of a participatory dental health education (DHE) programme for preventing early childhood caries (ECC). DESIGN: A one-year intervention programme. SETTING: 21 health centres. PARTICIPANTS: 520 mothers/caregivers of 6-19 month-old children who lived in a rural area of Thailand. INTERVENTION: Small group discussion with active involvement in the intervention group and the national teaching DHE programme in the control group. MAIN OUTCOME MEASURES: Health centre staff impact evaluation, children's dental cavitated carious increment and stated changes in oral health behaviour. RESULTS: After one-year, the percent of subjects using a toothbrush and tooth brushing with fluoride toothpaste was 93% and 87% respectively in the intervention group, significantly higher (p<0.01) than the control group (73% and 58% respectively). Night time bottle-feeding, falling asleep with a bottle and sweet snack diet behaviour appeared the same in both groups. The net cavitated carious increment was 3.5 (SD=3.4) teeth in the intervention and 3.2(SD=3.5) in the control group. Health centre staff were very supportive of the programme and suggested extending the participatory format to other child health topics. CONCLUSIONS: The participatory dental health education model was shown to be a practical and effective method for increasing oral hygiene practice, but was not sufficient to prevent the development of ECC. This single intervention in the short term is not seen as sufficient to prevent the development of ECC. PMID- 16379165 TI - Factors contributing to the variation in oral hygiene practices and facilities in long-term care institutions for the elderly. AB - OBJECTIVE: This study was designed to explore variations in oral hygiene practices and facilities in long-term care institutions for elders. Reported level of caregivers' knowledge related to oral health, reported work-climate, management, size of the institution and the mean age and degree of dependency of residents were evaluated. METHODS: Sixteen nursing homes were selected, using stratified random sampling, in the region of Gent (Flanders). Nine different strata were used based on size and management. Factors thought to be associated with the variation in oral hygiene practices and facilities were collected from 225 caregivers (75%) through a structured 45-item questionnaire. The questionnaire was validated and tested for reliability during a test-retest prior to the start of the study. The relation between rates of oral hygiene practices and facilities and explanatory variables was investigated using correlation analysis and subsequently by multiple regression analysis. RESULTS: The best fitted regression model explained 30% of oral hygiene practice variation. The most predictive variable was the knowledge of caregivers, which explained 17% of the variation, while mean age explained 7% and managerial behaviour 6.3%. There was a trend towards a negative relationship with mean age, degree of dependency and size of the institution. CONCLUSION: Most of the variance in oral health practices and facilities in long-term care institutions for elders in Gent remains unexplained. Knowledge, mean age and managerial behaviour were the most likely explanatory variables. PMID- 16379166 TI - Prevalence of malocclusion in 13-15 year-old North Jordanian school children. AB - OBJECTIVES: This study was carried out to determine the prevalence of malocclusion in 13-15-year-old North Jordanian schoolchildren. METHODS: One thousand and three children aged 13-15 years (619 females, 384 males) were examined using Bjork et al. (1964) registration method. A full clinical examination was carried out in the school premises using a mouth mirror under natural lighting. Alginate impressions were then taken for each student together with a wax bite. The impressions were poured the same day by an orthodontic technician. Peri-apical radiographs were taken for the students when hypodontia was suspected. RESULTS: Overall, the prevalence of malocclusion was 92%. Class II and III molars were found in 18.8% and 1.4% of the sample respectively. The malocclusion traits detected were crowding (50.4%), midline shift (31.7%), spaced dentition (26.7%), increased overjet (24.7%), deepbite (16.9%), crossbite (6.8), abnormal tooth form (7.7%), median diastema (6.9%), missing teeth (6%), ectopic canines (5.9%), inverted incisors (5.2%), high frenal attachment (3.9%), openbite (2.9%), tooth impaction (1.8%), reversed overjet (1.9%), scissorbite 0.3%. There was no statistically significant difference between males and females in the overall prevalence of malocclusion and the occlusal traits investigated. However, some dental anomalies showed significant differences. CONCLUSIONS: Prevalence of malocclusion among north Jordanians is comparable with other populations. PMID- 16379167 TI - Fluoride intake and fractional urinary fluoride excretion of Colombian preschool children. AB - AIMS: The purpose of this study was to assess the total fluoride intake and the fractional urinary fluoride excretion (FUFE) relative to the customary daily fluoride (F) ingestion in preschool children between 48-59 months of age. DESIGN: Total fluoride ingestion, from dietary and toothpaste samples was determined in 120 young children, dwellers of four Colombian cities. A "duplicate plate" technique was used. In Colombia, table salt is fluoridated to a concentration of between 180-220 mg F/kg. Individual (n=96) FUFE values were calculated as the ratio between the total amount of F excreted in the urine and the total amount of F ingested, over a 24-hour period. RESULTS: The average daily F-intake was 0.098 mg F/kg/day; 95% C.I. = 0.085-0.111 mg F/kg/day. The proportion of fluoride ingestion from toothpaste to the total fluoride intake was higher than 66% in all cities. The average FUFE values of subjects from each of the four Colombian cities under study did not differ significantly (ANOVA; p >0.91). The average 24 hour FUFE value for preschool children was 0.33; 95% C.I. = 0.29-0.37. CONCLUSIONS: The results obtained suggest that preschool children residing in Colombian urban areas are ingesting amounts of fluoride above the upper limit of the proposed safe threshold. FUFE values are similar to those reported in previous studies where daily F-doses were equal or higher than 0.064 mg F/kg. PMID- 16379168 TI - Enamel opacities in 8-year-old Icelandic children in relation to their medical history as infants. AB - OBJECTIVES: Opacities in tooth enamel were found in 34% of 8-year-old Icelandic children in 1970. This study aimed to measure the current prevalence of non fluorotic enamel opacities in the maxillary incisors and to determine if their occurrence was related to significant events in the medical history. DESIGN: The prevalence of demarcated enamel opacities was recorded photographically by calibrated examiners. SETTING: Subjects lived in Reykjavik, Iceland. SAMPLE AND METHODS: A random sample of 290 children aged eight years was examined in 1997-8. Parents were asked if the child had a history of (i) colic as an infant and/or (ii) repeated middle-ear infection. RESULTS: Demarcated white enamel lesions, not resembling fluorosis, were seen in 41% of children (teeth photographed wet) rising to 51% (teeth photographed dry). Enamel hypoplasia was seen in 11% of wet teeth and 15% of dry teeth. Parents of 94/288 children (32.6%) reported that their child had infant colic and 52/94 (55.3%) had received medication. Three episodes or more of middle-ear infections per year were reported for 123/290 (42.4%) children. Significant association was seen between the presence of demarcated enamel opacities and a history of otitis media. Diffuse enamel opacities, assumed to be fluorosis, were not associated with events reported from the medical history. CONCLUSIONS: Non-fluoride opacities of teeth are still prevalent in Icelandic children especially those with a history of infections in infancy. Without a careful diagnosis these opacities might be confused with fluorosis. PMID- 16379169 TI - Factors influencing referral of children for dental extractions under general and local anaesthesia. AB - OBJECTIVE: To investigate the relative importance of a range of explanatory variables concerning why child patients in Scotland enter pathways for tooth extractions under either Dental General Anaesthesia (DGA) or local anaesthesia (LA). BASIC RESEARCH DESIGN: A cross-sectional study was carried out involving DGA centres across Scotland. Data collected related to demographic characteristics of child DGA patients, reported anxiety and dental attendance levels of patients and parents, reasons given by referring practitioners for requesting DGA, number and type of teeth extracted and parental beliefs and attitudes to DGA. Similar data were collected relating to children having teeth extracted in primary care under LA. Multivariate analysis was performed to determine which factors were the best predictors of anaesthetic choice. RESULTS: 425 and 121 children having dental extractions under DGA and LA respectively participated. Ninety-six percent of DGA cases and 48% of LA patients had extractions for caries. For DGA cases, the mean age was 6.7 years, an average of 5.3 teeth were extracted and dental anxiety levels were higher than population norms. Multivariate analysis found the number of teeth extracted was the major predictor of anaesthetic type. On removing this variable, age became the most important factor, with 99% of children under 5.5 years receiving a DGA. Among DGA patients, 25% were reported to have had a previous DGA for tooth extraction. CONCLUSIONS: Future reductions in DGA numbers will be dependent on decreasing the number of young children presenting with advanced caries in multiple teeth. Consequently, guidelines are required concerning the prevention and appropriate management of caries in the primary dentition. PMID- 16379170 TI - Merits of financing systems influencing professional decision making. PMID- 16379171 TI - Influenza vaccination in Mississippi, 1992-2003: trends, subgroup comparisons, and forecast. AB - OBJECTIVES: We examined longitudinal trends in relative frequency of influenza vaccination in the state of Mississippi. METHODS: An influenza vaccination questionnaire was used to assess status among 20,078 adults who were sampled in random-digit-dialed telephone surveys administered in 1992, 1993, 1995, 1997, 1999, 2001, 2002, and 2003. Time trends were evaluated using logistic regression models in which the status of influenza vaccination was regressed on the respondents' gender, race, education, and household income. RESULTS: In Mississippi, influenza vaccination rates increased substantially during 1992-2003 irrespective of gender, race/ethnicity, age, education, and household income. The increases showed a significant linear trend. Non-Hispanic blacks were significantly less likely to have been vaccinated for influenza compared to non Hispanic whites without accounting for gender, age, education, and household income. Having a college degree was associated with higher vaccination rates among older Mississippians after adjusting for other factors. CONCLUSIONS: Although vaccination coverage is increasing, the national goal for 2010 does not appear achievable without new initiatives. Meanwhile, we should continue to provide funding for the immunization program and to educate the population regarding their compliance. PMID- 16379172 TI - Airhead. PMID- 16379173 TI - Did you wash your hands? PMID- 16379174 TI - Sick role is appropriate for hurricane victims as well. PMID- 16379175 TI - Subject: resolution no. 10-CME for community clinic service. Point. PMID- 16379176 TI - Subject: Resolution no. 10-CME for community clinic service. Counterpoint. PMID- 16379177 TI - Rubella immunization in Malaysia--20 years on, and the challenges ahead. PMID- 16379178 TI - Congenital deafness: high prevalence of a V37I mutation in the GJB2 gene among deaf school children in Alor Setar. AB - Twenty percent of all childhood deafness is due to mutations in the GJB2 gene (Connexin 26). The aim of our study was to determine the prevalence and spectrum of GJB2 mutations in childhood deafness in Malaysia. We analyzed the GJB2 gene in 51 deaf students from Sekolah Pendidikan Khas Alor Setar, Kedah. Bidirectional sequencing indicates that 25% of our childhood deafness has mutation in their GJB2 gene. Sixty two percent of these children demonstrate V37I missense mutation. Interestingly, V37I mutation in the GJB2 gene have been reported as polymorphism in Western countries, however in our country it behaved as a potentially disease-causing missense mutation, causing childhood deafness as it was not found in the normal control. PMID- 16379179 TI - Review of risk factors associated with the anastomosis leakage in anterior resection in Hospital Universiti Kebangsaan Malaysia. AB - This is a retrospective study evaluating the risk factors for clinical anastomotic leakage after anterior resection in 64 patients with rectal cancer operated from November 2001 till August 2003. The percentage of anastomosis leakage was higher in those patients with diabetic, low albumin level, neoadjuvant radiotherapy, higher staging, and poorly differentiated tumour. However, these differences were statistically not significant. There was a significant difference (p=0.03) of anastomosis leakage in those patients with very distal tumour of < 4cm from anal verge (42%), when compare to those patients with proximal tumor of > 15cm from anal verge (4.3%). The mortality associated with this complication was 30%. Hence, a surgeon may consider a covering stoma or be more vigilant in operating patients with multiple risk factors. PMID- 16379180 TI - Challenges in the treatment of sinonasal undifferentiated carcinoma: a ray of hope. AB - We studied nine cases of SNUCs presented to the Department of Otorhinolaryngology, Hospital University Kebangsaan Malaysia from 1999 to 2003. There were 8 males and 1 female with ages ranging from 24 to 78 years (mean 46.5y). The racial distribution consisted of 5 Chinese (55.5%), 3 Malays (33.3%) and 1 Indian (11.1%). Three patients were Kadish B (33.3%) and six were Kadish C (66.6%) by classification. In our series 2 years survival was 26.3% and median survival time was 14.2 months. PMID- 16379181 TI - Attitude towards rape: a comparative study among prospective physicians of Malaysia. AB - The global statistics reveal that at least one in every five women experiences rape or attempted rape during her lifetime. Rape myths encompass a set of (false) beliefs. Adolescents have high rates of rape victimization than other age groups. Rape myths among health care providers may have a negative influence on proper care of the victims. A total of 422 medical undergraduates of both sexes, studying at two Malaysian Institutes took part in the study. A validated questionnaire used in an earlier study was used for the present study, with a slight modification of scoring system. The age range of medical students was 17 to 34 years: mean +/- SD of men and women, were 20.6 +/- 2.1 and 20.3 +/- 2 respectively. Only about 19% of women and 11% of men had a very good positive attitude. Nearly 1/3 of women and 1/2 of men had a more negative attitude. On the whole the average total score of women was significantly higher (p=0.0004) than men. Nearly 50% of candidates with a village background carried more negative attitude. Violence against women is a significant public health problem. The major revelation of the present study is that only less than 20% of the medical undergraduates had a more positive attitude towards rape. Introduction of courses on 'sexual violence' in medical curriculum is likely to increase a) awareness, b) skills in management, c) in providing support and care for the victims and d) in implementing preventive actions. PMID- 16379182 TI - Prevalence of nocturnal polyuria in patients with benign prostatic hyperplasia. AB - Nocturia is a common symptom associated with benign prostatic hyperplasia which can persist even after effective treatment of BPH. Other causes may be responsible. We investigated a group of patients who were treated for BPH and continue to have nocturia. Our study has found a high prevalence (85.4%) of nocturnal polyuria in this group of patients. It has important clinical implication since the condition can be effectively treated with oral desmopressin. Empirical treatment without a routine frequency volume chart may be appropriate due to its high prevalence. PMID- 16379183 TI - Deaths in children during an outbreak of hand, foot and mouth disease in Peninsular Malaysia--clinical and pathological characteristics. AB - From July through December 1997, 11 previously healthy children in Peninsular Malaysia succumbed to an illness clinically characterised by an acute severe refractory left-ventricular failure, following a brief prodromal illness, in the midst of an outbreak of hand, foot and mouth disease (HFMD), similar to the reported experience in Sarawak and Taiwan. Retrospective reviews of the clinical features and results of laboratory, pathological and virological investigations of cases were conducted. The median age of the 11 case-patients was 31 months (range, 13 to 49 months); 6 were males. A brief prodromal illness of 3 days (range, 2 to 5 days) was characterised by fever (axillary temperature > 38 degrees C) (100%), oral ulcers (72%), extremity rashes (45%) and significant vomiting (55%). Upon hospitalisation, 7 of 11 case-patients had features suggestive of cardiogenic shock, while 4 of 11 case-patients developed shock during hospitalisation as evidenced by marked sustained tachycardia (heart rate > or = 180 beats per minute), poor peripheral pulses and peripheral perfusion, mottled extremities, pulmonary oedema (haemorrhagic pulmonary secretions in 8 of 11 cases during tracheal intubation, often precipitated by conservative crystalloid boluses, and radiographic evidence of acute pulmonary oedema in 5 of 7 cases) and markedly impaired left ventricular function on echocardiographic examination (7 of 7 cases). Three of 4 case-patients had aseptic meningitis while one case-patient also had an acute flaccid paraparesis. Despite supportive therapy, death occurred within a median of 13.4 hours following hospitalization. Post-mortem findings (all 8 specimens examined) consistently demonstrated brain stem encephalitis with foci of neuronal necrosis and micro-abscesses. None of the 11 specimens examined revealed histological evidence of myocarditis. Enterovirus 71 (EV71) was detected in 10 of 11 case-patients, many (7) from various sterile tissue sites (5 from central nervous tissues). No other viruses were isolated or identified. Clinical features and pathological studies closely paralleled the reported experience in Sarawak and Taiwan. The uniform necropsy findings of necrotizing brain-stem encephalitis coupled with essentially normal myocardial histology, in concert with the concurrent and consistent detection of EV71 points to a primary EV71 encephalitis; as yet unclear neurogenic mechanisms may account for the cardiovascular manifestations. PMID- 16379184 TI - Maternal passive smoking and its effect on maternal, neonatal and placental parameters. AB - A study was undertaken to 1) determine the effects of tobacco smoke exposure on maternal and neonatal weight and body mass index (BMI) and placental weight, volume and surface area and 2) establish any correlations between the placental surface area, volume and weight with maternal and neonatal body weight and BMI in mothers exposed to cigarette smoke. A total of 154 full-term placentae, 65 from mothers exposed to tobacco smoke and 89 from non-exposed mothers were collected from Kuala Lumpur Maternity Hospital. The placental surface area was determined using a stereological grid, the volume by Scherle's method and the weight by using an electronic weighing machine. In general there were no differences in maternal, placental and neonatal parameters between the exposed and non-exposed groups. However, there were significant correlations between placental weight with maternal weight and maternal BMI in both exposed (r = 0.315; p = 0.013) and (r = 0.265; p = 0.038), and non-exposed (r = 0.224; p = 0.035) and (r = 0.241; p = 0.023) mothers. It was also found that the maternal weight on admission correlated significantly with placental weight in both Malay (r = 0.405; p = 0.020) and Indian (r = 0.553; p = 0.050) passive smokers. Correcting the placental parameters for the maternal weight had no effect on the results. PMID- 16379185 TI - The use of mycophenolate mofetil in treating patients with non responding aplastic anemia. AB - Aplastic anemia is a relatively uncommon disease and conventional management options include immunosuppressive drugs and/or haematopoeitic stem cell transplantation. It is now known that the pathogenesis of aplastic anemia is immune mediated. Mycophenolate mofetil is a common immunosuppressive drug now used mainly in prophylaxis of graft rejection in organ transplant and also for prevention/treatment for graft versus host disease in haemtopoeitic stem cell transplantation. It is thought that mycophenolate mofetil may be useful in this group of patients. In this short report, mycophenolate mofetil was tried in 6 patients who had severe aplastic anemia with variable doses for a minimum duration of 9 months. The result has however not been encouraging. PMID- 16379186 TI - "Discovery to treatment" window in patients with smear-positive pulmonary tuberculosis. AB - Delay in commencing treatment in patients diagnosed with smear-positive pulmonary tuberculosis (PTB) may promote the spread of PTB in the community. Socio demographic and clinical data from 169 patients (119 retrospectively and 50 prospectively collected) treated for smear-positive PTB in our hospital Chest Clinic from June 2002 to February 2003 were analysed. One hundred and fifty eight (93.5%) patients were started on treatment in less than 7 days from the time when the report first became available while 11 (6.5%) patients had their treatment started > or = 7 days. The median 'discovery to treatment' window was 1 day (range, 0 to 24 days). Of the factors studied, longevity of symptoms, absence of fever or night sweats and having sought traditional medicine were associated with delay in treatment commencement. The urgency and importance of anti-TB treatment should be emphasized especially to patients who are inclined towards treatment with traditional medicine. PMID- 16379187 TI - Dialysis for end stage renal disease: a descriptive study in Penang Hospital. AB - This research was carried out to study the characteristics of ESRD patients and the resources consumed with dialysis treatment as well as to assess the clinical outcomes of ESRD treatment in Penang Hospital. A total of 117 ESRD patients were studied over 30 months. 56.4% of the patients were male and the median age was 40. Diabetic nephropathy was the commonest cause of ESRD (29.9%). Hypertension was the predominant comorbidity (60.6%). A larger proportion started treatment with Continuous Ambulatory Peritoneal Dialysis (59.0%). At the end of the study period, 49.6% of the patients continued their treatment in the same modality and 27.4% died. Average Dialysis Adequacy (Kt/V) achieved was satisfactory, 2.29 in CAPD and 1.50 in Haemodialysis. PMID- 16379188 TI - Does clinical exposure to patients in medical school affect trainee doctors' attitudes towards mental disorders and patients?--A pilot study. AB - Stigmatising attitudes towards mental illness may improve with clinical exposure during medical school training. Attitudes of 48 fourth year medical students in Universiti Kebangsaan Malaysia were assessed before and after their compulsory attachment in Psychiatry, using the Attitude Towards Psychiatry-30 (ATP) and the Attitude towards Mental Illness (AMI) questionnaires. ATP scores improved significantly with training (104.8 and 114.4, pre- and post-attachment respectively) as did AMI scores (63.4 and 68.1 respectively). Both improvements were statistically significant. (ATP: z = 4.55, p < 0.0005) (AMI: z = 3.75, p < 0.0005). Attitudes towards mental illness appeared to have become more favourable with clinical training in psychiatry. PMID- 16379189 TI - Turn-around-time of radiographs in a neonatal intensive care unit. AB - A quality assurance study was carried out prospectively in two phases at the Neonatal Intensive Care Unit (NICU) of Hospital Universiti Kebangsaan Malaysia. The objectives of the study were to determine the turn-around-time (TAT) of radiographs requested for infants undergoing intensive care treatment in the NICU and the effects of a standard operating procedure introduced based on initial findings of first phase of the study on subsequent TAT. The TAT was defined as the time taken for the radiograph to be ready for viewing after the attending doctor had requested for it to be done on an infant. During phase one of the study, none of the requested radiographs was ready to be viewed by the doctors within the standard TAT of 45 minutes. The problems identified were ward staffs delay in sending request forms to the radiology department, radiographers' delay in shooting and processing the films, and delay by NICU porter in collecting the processed films. Based on these findings, a standard operating procedure (SOP) was drawn up jointly by the staff of NICU and Department of Radiology. During phase two of the study conducted at one month after implementation of the SOP, there was a reduction of TAT by 50%. However, only 3 (4.3%) of the radiographs achieved the standard TAT. The main problems identified during phase two were delay in sending request forms and in collecting processed radiographs by the porter system. The dismal TAT of radiographs in NICU was related primarily to human behaviour. Besides continuous staff education, replacement of the porter system with electronic system may improve the TAT. PMID- 16379190 TI - Rubella vaccination programme in Malaysia: analysis of a seroprevalence study in an antenatal clinic. AB - In many developed countries, the incidence of rubella and congenital rubella syndrome (CRS) is considered to be negligible due to the availability of an effective vaccine. However, in Malaysia, several CRS cases are seen every year. This casts doubt on the effectiveness of the rubella vaccination programme. Very few seroprevalence studies were done over the years, making it difficult to discuss the effectiveness of the vaccination programme. The objective of this study is to determine the prevalence of rubella immunity among pregnant women attending antenatal clinics in a local teaching hospital. The hospital database on rubella immunity was assessed retrospectively from August 2001 to June 2002. A cross-sectional study of interviewed method as well as determination of rubella immunity by laboratory tests were carried out in July 2002. A total of 414 women were included, of whom 134 women were interviewed. The rubella immunity status was 92.3%. Based on this figure, rubella vaccination programme in Malaysia is a success despite the presence of CRS cases. Malaysia must ensure rubella vaccine coverage among target groups is high in order to minimise CRS cases. PMID- 16379191 TI - The role of climate on Malaria Incidence Rate in four governorates of Yemen. AB - The aim of study was to investigate the role of climate on the Malaria Incidence Rates (MIR) in some regions in of Yemen. For such purpose, the monthly (MIR) were calculated from the records of the hospitals' laboratories and centers of the Malaria Rollback centers in the main cities of the governorates Hudeidah, Taiz, Sana'a and Hadramout for the period 1989-1998. The readings of the climatic factors (CF) particularly the average monthly temperature (T), relative humidity (RH), volume of rain fall (RF) and wind speed (WS) for the same period of time were also collected from different weather and climatic information resources. Descriptive statistics, simple linear regression and multiple linear regression techniques were used to analyse the relationship between MIR and CF. The analysis shows highly significant relationship between MIR and the CF in these regions of Yemen (p-value 0.001). PMID- 16379192 TI - Asymptomatic chronically retained gauze in the pelvic cavity. AB - Cases of retained surgical guaze are rarely reported because of medico-legal and professional implications. Retained gauze for a period of more than 10 years is even rarer. A case of retained surgical gauze for 11 years, an accidental finding from a plain radiograph of a patient who had sustained proximal femoral fracture, is reported. A review of other reported cases is presented. PMID- 16379193 TI - Molecular genetic analysis of a supratentorial haemangioblastoma in a non-Von Hippel Lindau patient. AB - We describe a rare tumor site in a 46 year old man who presented with a two week history of headache. Physical examination revealed bilateral papilloedema with no other localizing signs. Computed Tomographic Scan as well as Magnetic Resonance Imaging of the brain revealed a lesion with a dura tail located adjacent to the falx cerebri of the right frontal lobe. This lesion was not invading the inner table of the skull base. A tumor blush was seen on angiogram. There were no abnormalities on CT scan of the abdomen and fundoscopy was normal. Intraoperatively a vascular tumor not attached to the dura was noted and removed totally. Histopathological examination was typical of a hemangioblastoma. Analysis revealed no mutations of the VHL gene in 5 regions, exon 5-8 of the p53 gene, exon 1-2 of the p16 gene and exon 5,6 and 8 of the PTEN gene. This is the first case report of a supratentorial hemangioblastoma in a non-Von Hippel Lindau patient with genetic evidence. PMID- 16379194 TI - Use of combined contralateral deltopectoral skin flap and ipsilateral island latissimus dorsi myocutaneous flap for massive cervico-thoracic oesophagus defect repair. AB - Restoring the intestinal continuity of an acquired massive cervico-thoracic oesophagus defect is a reconstructive challenge. A case requiring such defect restoration following a failed pedicled colonic interposition bypass graft between the cervical oesophagus and stomach for an intra-thoracic oesophageal perforation is presented. The defect between the oesophagostome at the lower left neck and the stoma of the colonic stump at the lower left chest measured about 20 cm. An ante-thoracic skin-tube neo-esophagus was constructed in two stages using a pedicled contralateral right deltopectoral skin flap and a pedicled ipsilateral island left latissimus dorsi myocutaneous flap (LD MC flap). A normal swallowing mechanism was re-established. PMID- 16379195 TI - Intractable epistaxis secondary to a post-traumatic pseudoaneurysm. AB - Epistaxis is a common problem encountered in clinical practice. It is usually self-limiting and is usually controlled with conservative measures such as nasal compression or ice-packs. Occasionally nasal packing is required. It is rarely severe enough that surgical intervention is warranted. The following report illustrates a patient who presented to us with a rare cause of life-threatening epistaxis that is, a post-traumatic pseudoaneurysm who finally required surgical intervention to control the bleeding. PMID- 16379196 TI - Bilateral anterior chamber intraocular lenses dislocation in a patient with habitual eye rubbing. AB - A 61-year-old Chinese man presented with bilateral posteriorly dislocated anterior chamber intraocular lenses (AC IOLs) one year after successful vitrectomy, removal of bilateral dislocated mature cataractous lenses and AC IOLs implantation. A thorough clinical evaluation revealed habitual eye rubbing as the only possible cause. PMID- 16379197 TI - Kimura's disease: an unusual cause of cervical tumor. AB - Kimura's disease (KD) is an angiolymphoid proliferative disease of soft tissue with peripheral blood eosinophilia and elevated serum immunoglobulin (Ig) E. The treatment options range from conservative observation for the asymptomatic patient to surgical excision, steroid therapy and radiotherapy for symptomatic patients. Surgical excision is the most common diagnostic measure and is the treatment of choice. A case of KD in a 13-year-old Malay girl is presented. Clinically there was painless right jugular digastric mass measuring 3cm by 3cm. Her blood investigation showed pronounced eosinophilia. She underwent excision biopsy uneventfully. The biopsy from the swelling showed reactive follicular hyperplasic with prominent eosinophilia. There was no evidence of malignant change. Postoperatively after 3 years follow up, she was asymptomatic and no signs of tumor recurrence. PMID- 16379198 TI - Delayed complication of radiotherapy: laryngeal fibrosis and bilateral vocal cord immobility. AB - Radiotherapy is the accepted treatment for early laryngeal carcinomas with the advantage of voice preservation. It is however, not without complications. We report a case of bilateral vocal cord immobility 15 years after radiotherapy. This appears to be due to ankylosis of cricoarytenoid joint and fibrosis of the larynx. PMID- 16379199 TI - Cochlear implantation in congenital cochlear abnormalities. AB - Many children have benefited from cochlear implant device including those with congenital malformation of the inner ear. The results reported in children with malformed cochlea are very encouraging. We describe 2 cases of Mondini's malformation with severe sensorineural hearing loss. Cochlear implantation was performed and both of them underwent post-implantation speech rehabilitation. Post-implantation, both of them were noted to respond to external sound. But the second case developed facial twitching a few months after the device was switched on. It is important to evaluate the severity of the inner ear deformity and the other associated anomalies in pre-implantation radiological assessment in order to identify the problem that may complicate the surgery and subsequent patient management. PMID- 16379200 TI - Migration of a foreign body in the maxillary sinus illustrating natural mucociliary action. AB - We present a case of a foreign body which migrated to the maxillary ostia by mucociliary action from its initial location on the floor of the maxillary sinus where it was traumatically introduced. This report illustrates that a powerful mechanism of mucociliary action can cause relatively heavy objects within the maxillary sinus to migrate naturally to the sinus ostia against gravitational force. PMID- 16379201 TI - Histoplasmosis of the larynx. AB - Primary histoplasmosis of the larynx is not a common disease. Most cases of laryngeal histoplasmosis results from hematogenous spread of disseminated histoplasmosis usually originating from pulmonary infection by Histoplasma capsulatum. We report a 52-year-old male chronic smoker who had prolonged hoarseness and was initially diagnosed with laryngeal carcinoma. However, biopsy of the laryngeal mucosa confirmed the diagnosis of histoplasmosis. There were no signs of pulmonary or systemic involvement. Treatment with intravenous amphotericin B was given for a week and oral ketaconazole was given for a month with complete resolution of symptoms. PMID- 16379203 TI - Potential problems with interventions that restrict access to patients who present inappropriately to Emergency Departments. PMID- 16379202 TI - Mycoplasma pneumoniae infection complicated by empyema: a rare presentation. AB - Mycoplasma pneumoniae is a common causative agent for childhood pneumonia. However, empyema is a rare presentation. We report a case of a previously well child who presented with a right-sided empyema. M. pneumoniae was confirmed serologically with evidence of a four-fold rise in Mycoplasma IgM titre. The empyema required drainage procedures for more than two weeks. The infection resolved with a course of six weeks of treatment with erythromycin. PMID- 16379204 TI - Understanding hyperemesis gravidarum. AB - Nausea and vomiting are common in early pregnancy affecting 70-80 percent of pregnant mothers. In a majority of women vomiting begins between 4-7 weeks of pregnancy. Nausea and vomiting are usually mild and self-limiting, however some of the mothers have a more profound course which lead to hyperemesis gravidarum. Careful clinical evaluation is necessary to exclude underlying medical illnesses or non pregnancy related causes of severe vomiting. Hyperemesis gravidarum poses health risk to both mother and baby, therefore prompt treatment should be initiated without delay. Non pharmacotherapy such as dietary modification and emotional support are useful. Pharmacotherapy with antiemetics, pyridoxine, methylprednisolone are effective and relatively safe. Severe hyperemesis with dehydration and electrolyte imbalance may need hospitalisation for electrolyte and fluid replacement. PMID- 16379205 TI - Evaluation of the outcomes of 7 Class II treatments 40 years later. AB - AIM: To show that starting treatment of Class II malocclusions around the time of peak height velocity at adolescence enables the orthopedic and fixed appliance steps to be merged into a single continuous phase, leading to good outcomes with long-term stability. METHODS: Seven patients with Class II malocclusions began treatment in the mid- 1960s with cervical headgear and bite plate, followed immediately by fixed appliances. Photographs and study casts were obtained for the same 7 patients approximately 40 years later. RESULTS: The stability of individual cases was considered to be good over the 40-year period. Crowding changes in the mandibular incisor region were minimal. CONCLUSION: A two-step single-phase treatment for Cass II malocclusions during adolescence has proved to be an efficient approach throughout many years of orthodontic practice. Furthermore, by using ossification markers, the individual child's growth potential can be estimated. This knowledge leads to greater efficiency by developing a positive, purposeful, and more confident approach to the management of Class II cases. PMID- 16379206 TI - Evaluation of stomatognathic function in orthodontic treatment. AB - AIM: To evaluate changes in stomatognathic function related to orthodontic treatment in 4 patients with various types of dysfunction of the stomatognathic system present prior to orthodontic treatment. METHODS: Four patients who visited the Orthodontic Clinic at Tokyo Medical and Dental University Hospital and whose active orthodontic treatment was complete were analyzed both morphologically and functionally. To analyze stomatognathic function, electromyographic activity of the masticatory muscles and mandibular movements was recorded. RESULTS: In all 4 patients, changes in stomatognathic function were observed following improvements in craniofacial structural and interocclusal relationships. Many of these changes consisted of elimination of functional disharmony, although the cause-effect relationship between these functional changes and orthodontic treatment is unclear. CONCLUSION: Functional analysis during orthodontic treatment can provide useful information concerning the relationship between craniofacial structures and stomatognathic function. PMID- 16379207 TI - Planas direct tracks in young patients with Class II malocclusion. AB - In Brazil, Class II malocclusions affect approximately one-third of children in the primary dentition period, and approximately two-thirds of the adolescent population. According to many authors, this type of malocclusion worsens with time, due to facial growth during childhood, both in terms of quantity and quality, and the facial pattern is established at an early age. The application of the Planas Direct Tracks concept and technique may represent an interesting tool for the correction and prevention of Class II malocclusion in an early treatment approach, working 24 hours a day, 7 days a week, applying oral functions and muscle activity to correct the malocclusion. PMID- 16379208 TI - The congenitally missing maxillary lateral incisor. Part 1: esthetic judgment of treatment options. AB - AIM: To determine how general dentists, orthodontists, combined dental specialists, and laypeople judged the relative attractiveness of a series of photographs of teeth that included cases with congenitally missing incisors to compare the esthetic appeal of various treatment options. METHODS: The photographs included cases with Maryland bridges, dental implants, and orthodontic substitution of the lateral incisors with the canines. Cases with no missing teeth were included as controls. The subjects rated each photograph independently, assigning a number between 1 and 5 for each of a series of 7 bipolar adjectives. Each photograph was given an overall score from 7 (best) through 35 (worst) for use in statistical analysis (ANOVA). RESULTS: Results indicated that general dentists, orthodontists, combined dental specialists, and laypeople differed in their evaluations of the photographs, which represented multiple treatment options for congenitally missing maxillary lateral incisors. CONCLUSIONS: Including esthetics of the finished result as an advantage or disadvantage for either restoration or canine substitution of the lateral incisors could be misleading to the patient because the judgment of esthetics of a smile can vary from one person to another or from one dental professional to another, as indicated in this study. PMID- 16379209 TI - The congenitally missing maxillary lateral incisor. Part 2: assessing dentists' preferences for treatment. AB - AIM: To (1) determine if there are different preferences among general dentists, a combined group of dental specialists, and orthodontists concerning whether a patient with congenitally missing lateral incisors should have the teeth restored or if the canines should be orthodontically moved into the position of the missing teeth; (2) determine the role that esthetics and function of the final result has on the respondents' treatment preferences; and (3) compare the respondents' opinions about treatment preferences to their rankings of esthetics reported in part 1. METHODS: The questionnaire accompanied the photographic evaluation in part 1. Percentages were computed and chi-squares were used to compare response patterns. RESULTS: Compared to orthodontists, a significantly greater percentage of general dentists and combined dental specialists would restore the lateral incisors and would do so primarily for esthetic reasons. Interestingly, for those dental professionals who responded that the missing teeth should be restored, many did not rank any photograph of a restorative option as the best option. CONCLUSIONS: Dental professionals should attempt to eliminate their personal opinions when recommending treatment options for a congenitally missing maxillary lateral incisor because discrepancies exist between the treatment result judged as most esthetic and the one most likely to be recommended. PMID- 16379210 TI - A comparison of two maxillary molar distalizing appliances with the distal jet. AB - AIMS: Previous studies on maxillary molar distalization have usually concentrated on only one appliance and featured small sample sizes. The purpose of this retrospective study was two-fold: (1) to determine the skeletal, dental, and soft tissue effects of 3 molar distalization appliances, 2 of which do not depend upon patient compliance (ie, distal jet and Greenfield molar distalizing appliance) and 1 that does (ie, sagittal appliance combined with cervical headgear); and (2) to determine differences in treatment effects among the 3 appliances. METHODS: Pretreatment and post-distalization cephalometric radiographs were obtained for each appliance (14 females and 11 males for the distal jet; 12 females and 13 males for the Greenfield molar distalizing appliance; and 17 females and 13 males for the sagittal appliance with headgear). RESULTS: Pretreatment to transition evaluation showed significant distal movement of the first molars for the distal jet (3.4 mm), the Greenfield molar distalizing appliance (3.9 mm), and the sagittal appliance with headgear (2.1 mm). Distal tipping of the first molar was seen in all samples, but significantly more so in the Greenfield molar distalizing appliance (6.5 degrees +/- 6.6) and the sagittal appliance with headgear (13.5 degrees +/- 8. 1) than in the distal jet (3.2 degrees +/- 2.8). CONCLUSIONS: Maxillary molar distalization was effective using the distal jet, the Greenfield molar distalizing appliance, and the sagittal appliance with headgear, but better control of molar bodily movement was reported with the distal jet. PMID- 16379212 TI - Incisal edge recontouring in orthodontic finishing. PMID- 16379211 TI - Morphologic skeletal asymmetry, with a Class III skeletal discrepancy, treated without surgical intervention. AB - Patient J.S., adult, female, presented with a morphologic skeletal asymmetry of the mandible resulting in a unilateral posterior crossbite, and a Class III skeletal discrepancy characterized by a hyperdivergent facial pattern (FMA, 30 degrees), retrognathic maxilla (SNA, 79 degrees), and compensating maxillary alveolar dental protrusion (1-NA, 7 mm). Maxillofacial surgery was declined. Orthodontic treatment, utilizing the Tweed-Merrifield edgewise technique, widened the maxillary dentition with a combination buccal crown torque and buccal root torque and with lingual crown torque in the mandibular posterior quadrant. Temporomandibular joint symptoms lessened and function improved with orthodontic treatment. The skeletal asymmetry improved and the patient was happy with the result, including her profile and her beautiful smile. PMID- 16379213 TI - A rationale for expansion. AB - Over the past 100 years, orthodontists have vacillated between extremes of nonextraction and extraction therapies. Injudicious selection of therapies despite facial dimensions has probably contributed to the major clinical disappointments within each style of therapy. The belief that expansion distal to the canines would not stabilize after the cessation of active treatment has contributed to the reluctance to use such therapy. However, experience has shown that this type of expansive, nonextraction therapy can have success without relapse and merits more attention from the specialty. PMID- 16379214 TI - VIP interview. Julia Harfin. Interview by Samir E. Bishara. PMID- 16379215 TI - France: Fraternite in orthodontics. PMID- 16379216 TI - Conquering doubt. PMID- 16379217 TI - Preparing Kentucky's emergency departments for a comprehensive trauma care system: the Kentucky Hospital Association emergency department survey. AB - Kentucky continues to lag in development of a statewide trauma care system, and objections to implementation have arisen from hospital emergency departments. In order to clarify these issues, the Kentucky Hospital Association conducted a statewide survey. Respondents represented 95% of the state's general acute care hospitals. A large majority (78%) reported that they could not meet the American College of Surgeons' (ACS) standards for verification as a trauma care facility at any level, and a similar number (81%) opposed requiring verification for facilities providing trauma care. Staffing deficiencies (particularly surgical specialists) were the primary reason cited for facilities' inability to meet ACS standards, while objections to requiring ACS verification were based on concerns about the ability of EMS to transport patients to definitive care in a timely enough manner. Recommendations included improvements in emergency department (ED) facilities and staffing as well as EMS capacity and training. Improvements in Kentucky trauma care will require additional funding to increase the number of verified facilities and enhance coordination among providers and facilities. PMID- 16379218 TI - Kentuckians support trauma system improvement: the Kentucky Trauma Survey report. AB - A household telephone survey was conducted in mid-2004 to assess the degree of information and support among Kentuckians regarding the state's trauma care system. The 800 completed responses reflected a very strong degree of support (79%) for trauma system improvement and funding. Responses also indicated low levels of understanding regarding trauma care facilities and their level of service. Respondents with personal or family experience of emergency medical services and trauma care were significantly more likely to identify trauma facilities correctly. The most popular option (69.6%) for generating additional trauma system funds was a 5% levy on guns and ammunition. Three quarters (74.2%) of respondents indicated that they would vote for a legislative candidate who supported trauma system funding. Overall, survey findings demonstrate a strong, broad mandate for legislative initiatives to improve Kentucky's trauma care. PMID- 16379219 TI - Please help me die. PMID- 16379220 TI - An epic year for a great society. AB - As 2005 comes to a close, I have been asked to summarize the year's accomplishments and efforts at the County level. In many ways HCMS reminds me of "the little engine that could"-aided by a single, extremely dedicated and talented staffer and bolstered by an all volunteer army of selfless professionals with a focused Executive Committee at the helm, we have indeed "chugged up that big hill" in taking on a variety of traditional and not-so-traditional challenges. PMID- 16379221 TI - Comparing the teaching efficacy of a procedure-in-a-box toolkit to a live instructional workshop. AB - BACKGROUND: Emergency procedures have been traditionally taught in live "hands on" workshops, which are expensive (tuition, travel, hotel, and leaving one's practice idle). This study was conducted to compare the teaching efficacy of a traditional live workshop to a procedure-in-a-box toolkit (PBT) method which contains audiovisual instructions and props to practice the procedures. METHODS: Four procedures, zipper release, intraosseous needle placement, fishhook removal, and splinting, were taught to 32 physician volunteers, using both teaching methods. Each participant was asked to evaluate the teaching method after each session. RESULTS: The mean educational quality of each method were not significantly different from each other, but if given a choice study subjects preferred the live workshop more often. CONCLUSION: The live workshop is the preferable method for teaching procedures but when considering expenses, most of the subjects felt that the PBT method is an adequate substitute method for the live workshop. PMID- 16379222 TI - A case control investigation of hepatitis C risk factors in Hawaii. AB - This case control investigation of hepatitis C risk factors in Hawaii showed that IV drug use, blood transfusion, tattoos, incarceration, acupuncture, prior dental or oral surgery, and HIV infection were associated with HCV Future public health efforts in Hawaii should focus on developing effective and appropriate community interventions targeting those with well-established risk factors for HCV PMID- 16379223 TI - The educational approach to advanced cancer: a preliminary report. AB - Survival from cancer depends primarily on reduction of tumor burden through surgery chemotherapy and radiotherapy. But other factors such as energy levels, immunity and will to live have long been known to play a part. Some of these factors are best provided by complementary therapy. Our program, educational in nature, attempts to make these modalities available to the patient. PMID- 16379224 TI - Revolution and evolution in the Department of Surgery. PMID- 16379225 TI - A community-based approach to tobacco prevention: Hawaii's youth taking on the tobacco industry. PMID- 16379226 TI - ["Pneumology will take its place in this dynamic period". Rehabilitative medicine awakens hope]. PMID- 16379227 TI - [COPD: the smoking habit is the decisive preventive measure]. PMID- 16379228 TI - [Walking pneumonia: new guidelines say that the struggle starts in the lung]. PMID- 16379229 TI - [Bronchial carcinoma: does the solution come with individualized therapy?]. PMID- 16379230 TI - [Are we achieving the maximum solution in allergies and asthma? Interview by Roland Fath]. PMID- 16379231 TI - [Interventional cardiology. Techniques which involve the heart]. PMID- 16379232 TI - [Spontaneous idiopathic thrombosis: Guidelines for prevention have been established. Interview by Inge Kelm-Kahl]. PMID- 16379233 TI - [Cardiac insufficiency: strength for the weak heart]. PMID- 16379234 TI - [Cardiovascular risk factors--where do these come from? Interview by Nicola von Lutterotti]. PMID- 16379235 TI - [Chronic inflammatory intestinal diseases. When the intestinal mucosa becomes thin]. PMID- 16379236 TI - [Cancer of the large intestine. Interview by Felicitas Witte]. PMID- 16379237 TI - [Viral hepatitis: challenge for the politics of health]. PMID- 16379238 TI - [Medical guidelines: New guidelines for gastroesophageal reflux. Interview by Felicitas Witte]. PMID- 16379239 TI - [Transplantation medicine: issue of blood group incompatibility]. PMID- 16379240 TI - [Dialysis in Germany: new medications and new technical possibilities. Interview by Sonja Bohm]. PMID- 16379241 TI - [Rheumatology: "Today's therapy is a complete remission." Interview by Cornelia Stolze]. PMID- 16379242 TI - [Rheumatoid arthritis: earlier and more aggressive therapies]. PMID- 16379243 TI - [Antibiotics & Co: new strategies against infectious diseases]. PMID- 16379244 TI - [Infectious diseases: How big is the real danger? Interview by Strehlow Karen]. PMID- 16379245 TI - [Molecular biological strategies in cancer. How effective are the new therapies?]. PMID- 16379246 TI - [What happens after cancer in children?]. PMID- 16379247 TI - [National Center for Cancer in Heidelberg. Specific treatments for the needs of patients]. PMID- 16379248 TI - [Breast cancer: Needs in care and therapy]. PMID- 16379249 TI - [Prostate cancer: between early diagnosis and total care]. PMID- 16379250 TI - [National Diabetes Center: competence center for research and therapy]. PMID- 16379251 TI - [On the development of insulin: always more security and comfort for diabetics]. PMID- 16379252 TI - [Prevention of type 2 diabetes: "We need preventive measures". Interview by Rudiger Meyer]. PMID- 16379253 TI - [Type 1 diabetes mellitus: ripe for islet cell transplantation?]. PMID- 16379254 TI - [Therapies for overweight and obesity]. PMID- 16379255 TI - [Osteoporosis: What have the Osteology Society's guidelines done?. Interview by Wiebke Kathmann]. PMID- 16379256 TI - [Hormone replacement therapy. Interview by Wiebke Kathmann]. PMID- 16379257 TI - [Post cardiac surgery syndrome]. PMID- 16379258 TI - [Effect of overweight on liver function]. PMID- 16379259 TI - [Increased carcinoembryonic antigen: what is the diagnosis?]. PMID- 16379260 TI - [Systemic mastocytosis]. PMID- 16379261 TI - The Learning Academy. PMID- 16379262 TI - Nursing's pioneers in the associate degree movement. PMID- 16379263 TI - Debbie. PMID- 16379264 TI - Storytelling as cultural assessment. AB - This article describes how story was used to teach cultural assessment to baccalaureate nursing students enrolled in clinicals in long-term care facilities. Class sessions focused on learning to elicit and listen to patients' stories and use story as an assessment tool in clinical practice. With their peers, students learned to listen in a nonjudgmental, contextual way to the values and beliefs of the storyteller. They learned that all people, even those from the dominant mainstream culture, have stories to tell, and that stories build bridges between nurses and clients. PMID- 16379265 TI - After the funding is gone: evaluating the sustainability of a community-based project. AB - The Rural Elderly Enhancement Project was a nurse-initiated grant funded by the W.K. Kellogg Foundation and administered by an urban school of nursing. The major purpose of the nine-year grant was the development of a model of community participation and empowerment. An informal evaluation was conducted to assess the communities' perception of the project and to determine the sustainability of the project's initiatives. A random sample of individuals from community agencies, schools, and churches were interviewed by six senior baccalaureate nursing students enrolled in a community nursing course. Results demonstrated that the communities had a very positive view of the project and that many project initiatives were sustained and have expanded. PMID- 16379266 TI - Linking remote practice to research: technology in action. AB - Nursing faculty practices are rarely set up to collect and analyze data, but evidence about the effects and costs of clinical care provided by faculty is required by health plans and other payers. This article describes a strategy that links faculty research programs with practice and teaching activities in various locations. An informatics infrastructure uses a common language for data collection and a set of coherent processes for implementation. The theoretical basis for conducting practice-based research is explored. PMID- 16379267 TI - Addressing the community research needs of baccalaureate students. AB - This article describes a course designed to prepare RN-BSN students to assimilate relevant research findings into clinical practice. The course was developed as faculty accepted the challenge of providing didactic content completely online. Guided by two faculty members, two groups of students developed research proposals to respond to community needs identified by a local hospital and a nursing center. PMID- 16379268 TI - Student contributions to clinical agencies: a comparison of adult health and psychiatric staff nurses' perceptions. AB - Clinical rotations in acute medical and psychiatric inpatient facilities are an essential component of the education for undergraduate nursing students. The benefits students receive from these clinical experiences have been documented, but no data have been reported regarding the advantages that students may bring to agencies. This exploratory study, which used a mail-back survey, asked adult health/medical-surgical nurses and mental health/psychiatric nurses in acute care units about the effects undergraduate nursing students had on staff time, staff development, quality of care, staff's personal satisfaction, and unit standards and practices during their clinical rotations. Results indicated that students contribute to the personal and professional satisfaction of the staff by offering opportunities for mentoring and reciprocal learning and allowing nurses to participate in students' professional development. Students also contribute to patient care. PMID- 16379269 TI - [Calculation of spectroscopic parameters of highly doped Er3+ in lithium niobate]. AB - A highly doped Er3+: LiNbO3 (concentration 6 mol%) crystal was grown successfully by Czochralski method. The crystal is higher than that of the lowly doped Er3+ in LiNbO3 crystal, which is helpful to improve absorption coefficient of the grown the pumping efficiency. The absorption spectra at two unpolarized directions (X and Z) and two polarized directions (E parallel Z, E perpendicular Z) were measured. Using the Judd-Ofelt theory, and according to the measured absorption spectra, the intensity parameters omegalambda of Er3+ were fitted. The results of root-mean square (r. m. s) deviation show that the error of polarized fitting is less than that of unpolarized one. Thus fluorescence transition probabilities (Ajj), radioactive lifetime (tau), fluorescence branching ratio (beta), and integrated emission cross section (sigmap) were calculated and accepted according to the polarized results, and were also discussed and compared with the ones reported in the literature. PMID- 16379270 TI - [The upconversion mechanisms for the red and green emissions in Er3+ doped and Er3+ -Yb3+ codopoed fluoride materials based on experiment]. AB - The rare earth Er3+ doped and Er3+ -Yb3+ co-doped fluoride samples were prepared. Under 980 nm laser excitation, the intense red (650 nm) and green (545 nm) upconversion luminescence was obtained corresponding to the transitions 4S3/2, 2H11/2--4I15/2 and 4F9/2--4I15/2, respectively. For the Er3+ -Yb3+ co-doped fluoride samples, it was found that the ratio of the red upconversion luminescence to the green one decreases as the Er3+ concentration increases. The dependence of upconversion intensities on excitation power indicates that two photon absorption processes are responsible for the upconverted luminescence. The possible upconversion mechanisms of the red and green emissions are discussed based on the energy-matching conditions and the quadratic dependence on excitation power. PMID- 16379271 TI - [Infrared photostimulated luminescence and optical storage compares in alkaline earth sulphide doped with Mn]. AB - The phosphors SrS: Eu and CaS: Eu doped with Mn were prepared by sulfurization flux method (SFM). Infrared photostimulated luminescence (PSL) of both the phosphors were measured. The PSL peak wavelength of SrS: Eu, Mn is 610 nm. Comparing the storage intensities, SrS has better optical properties. The doping of Mn2+ increases photon amount and improves the storage properties. PMID- 16379272 TI - [Infrared and raman spectra studies of DL-homocysteic acid (DLH) and its lanthanum complex at high external pressure]. AB - DL-Homocysteic acid (DLH, (NH3 )-CH(COOH)-(CH2 )2-SO3-) is extensively in existence as free amino-acid in the central nervous system of mammal. The infrared (IR) and Raman spectra of DL-Homocysteic acid (DLH) and its La complex [La (DLH)2Cl3 . H2O = LaL2] were measured at high external pressure. The DLH has two pressure-induced phase transitions (near 17 kbar and 37 kbar) and three pressure-phase areas below 50 kbar for there are ten intermolecular hydrogen bonds between the DLH molecules. But there is only one pressure-induced phase transition (near 27 kbar) to be observed in the complex LaL2. These show that the ten intermolecular hydrogen bonds of the DLH molecules changed due to the formation of complex LaL2. In infrared spectra, the changing trend of the pressure sensitivities (dv/dp) of symmetry stretching of SO3- in DLH and asymmetry stretching of SO3- in LaL2 are very different from the other vibrational modes when both compounds undergo high external pressure. In infrared spectra, the average pressure sensitivities of symmetry stretching of SO3- are: low-pressure phase area (0.30 cm(-1) x (kbar)(-1)) < middle-pressure phase area (0.32 cmn(-1) (kbar) (-1)) < high-pressure phase area (0.41 cm(-1) x (kbar)(-1)), but 0.86 cm(-1) x (kbar)(-1) (low-pressure phase area) >0.64 cm(-1) x (kbar)(-1) (middle-pressure phase area) >0.26 cm(-1) x (kbar)(-1) (high-pressure phase area) to other vibrational modes in DLH. The average pressure sensitivities of asymmetry stretching of SO3- in LaL2 are lower in low-pressure phase area than in high-pressure phase area, but the average pressure sensitivities of other modes are higher in low-pressure phase area than in high-pressure phase area. PMID- 16379273 TI - [Investigation on the nitrogen functionality of volatile through FTIR spectroscopy equipped with a long path distance gas cell]. AB - Nitrogen functionalities of volatile of three different rank coals, namely TONGCHUAN lean coal, SHENMO bituminite and YIBIN anthracite, were studied by using Fourier transform infrared spectroscopy (FTIR) equipped with 16 m long path distance gas cell. The detection problem of low content of nitrogen in volatile was successfully solved because of the application of the long path distance gas cell. In the long path distance gas cell, the infrared penetrates the sample more times than in a normal gas cell. Results from the analysis of spectra obtained by FTIR show that there are four functionalities of nitrogen in volatile, including pyrrole-type nitrogen, pyrindine-type nitrogen, pyridine-N-oxide nitrogen, and nitrile nitrogen. Comparison of research results of nitrogen group between coal and volatile suggests that the difference in fate of nitrogen between volatile and coal results from the macromolecular nitrogen group of coal decomposition under the effect of temperature. PMID- 16379274 TI - [Determination of glycyrrhizic acid in glycyrrhiza uralensis fisch by fiber optic near infrared spectroscopy]. AB - The objective of this study was to develop a rapid, non-destructive and real-time method for the determination of glycyrrhizic acid in glycyrrhiza uralensis fisch. Near infrared spectra (NIR) in the range of 10 000-4 000 cm(-1) were recorded for the glycyrrhiza uralensis fisch containing glycyrrhizic acid in the content of 0.94 %-3.06%. Calibration models were established using the PLS (partial least squares) and PCR (principle component regression) algorithm. Different spectra pretreatments methods were compared. The study showed that PLS model gave better results than PCR with the correlation coefficient 0.958, SEC 0.179 (standard deviation of the calibration sets) and SEP 0.197 (standard deviation of the prediction sets). Results indicate that fiber optic NIR method can be used to on line control the valid component in Chinese herbs. PMID- 16379275 TI - [Applied study on support vector machine (SVM) regression method in quantitative analysis with near-infrared spectroscopy]. AB - This paper introduced the application of support vector machines(SVM) regression method based on statistics studytheory to the quantitative analysis with near infrared (NIR) spectroscopy. Sixty-six wheat samples were used as experimental materials, and thirty-three of them were used as calibration samples. The protein contents and NIR spectra of the calibration samples were used to build SVM regression models by four different kernel functions. The protein content of the predicting samples are estimated by four different SVM regression models. All of the correlation coefficients between the estimated values by different SVM regression models and the standard chemical values of protein content by Kjeldahl's method are more than 0.97. The average absolute error is less than 0.32. To investigate the predicting effect, it is compared with PLS regression models. The result suggested that the SVM regression, which was built to estimate the protein content of wheat samples, can also be used in the quantitative analysis of real samples by NIR. PMID- 16379276 TI - [Predicting the chemical composition of intact kernels in maize hybrids by near infrared reflectance spectroscopy]. AB - Intact-kernel samples of normal maize inbred lines and hybrids were collected from field experiments of three locations. Calibration equations were developed by partial least square regression (PLS) of chemical values of near infrared reflectance spectroscopy (NIRS) data and tested through both cross and external validation. In addition, 40 progenies of F1 and F2 generation not included in calibration and validation sets were verified to further evaluate the reliability of three calibration equations. The authors found the coefficients of correlation (r) of 0.98, 0.93 and 0.97 between NIRS predicted and actual protein, starch and oil content in these materials, respectively. However, the greatest relative errors were 2.7% (protein), 2.46% (starch) and 7% (oil). Thus, the accuracy of prediction could be comparable to chemical methods. The feasibility of developing NIRS equations with samples of inbred lines to determine grain quality of hybrids was also examined. The analysis of principal components of spectrum of the inbred lines and hybrids supported a new theory that plant spectrum properties could be heritable. PMID- 16379277 TI - [Characterization of the orientation of polymer material by non-polarized raman laser light spectroscopy]. AB - The non-polarized Raman laser light was used to investigate the anisotropic polymers including polypropylene fiber, nylon 6 flat fiber, and polyethylene pipe. The Raman spectra of the same samples with different location relative to the incident direction of laser beam show obvious differences. The present result extends the application of Raman spectroscopy to the characterization of oriented polymers samples. PMID- 16379278 TI - [Surface-enhanced Raman spectroscopy in studies of corrosion and inhibition on an iron surface]. AB - Application and advanced achievement of surface-enhanced Raman spectroscopy (SERS) in corrosion and inhibition on an iron surface have been reviewed. As a technology providing the molecular information of material structure, SERS is widely applied to the studies of metal corrosion and inhibition, especially for iron. Applications of this spectroscopy technology are showed in three aspects: the SERS enhancement theory of Fe surface, the adsorbent mode of inhibitions on Fe surface, and the structure of oxidation or passivation film on a Fe electrode. Furthermore, the tendency of farther study is prospected. PMID- 16379279 TI - [Surface-enhanced raman spectra studies on roughened Zn electrode in alkaline solutions]. AB - Electrochemical oxidation-reduction method was employed to roughen Zn electrode for obtaining SERS, and potential dependent surface enhanced Raman spectra (SERS) of roughened Zn electrode in KOH solution of different concentration wereobserved. The spectra of Zn electrode in various solutions had obvious differences which indicated the concentration of OH- had a great effect on the dissolution and passivation of zinc. Based on our experimental results, the authors attempt to analyse the behavior of zinc in alkaline and give the mechanism of its passivation. PMID- 16379280 TI - [Application of Raman spectra to the research of jades excavated from Xue Jiagang site]. AB - Non-destructive analysis is always the aim in jades' research. The present article applied the Raman spectroscopy to the research on jades excavated from the Xue Jiagang site and achieved good result in the main mineral, inclusion and phenocryst. The study shows that as a non-destructive technique Raman spectroscopy can be applied to ancient jades conveniently and practically, and it can detect not only the surface information of ancient jades, but also the interior information. The technique is important to the verification and provenance of ancient jades. PMID- 16379281 TI - [Experimental study of Raman spectra of magnesite at 297 K and at the pressure of 0.13-1 GPa]. AB - The experimental study of Raman spectra of magnesite has been conducted at the pressure of 967 MPa and at the temperatureof 297 K using a cubic zirconia-anvil cell. The result shows that neither phase transition nor organic substances were observed during compression, and the Raman peak of magnesite shifted to higher frequency with increasing pressure. The relation between the pressure and the Raman peak position of magnesite (1094 cm(-1)) was obtained as follows: v (cm( 1)) = 0.007 44 x P(MPa) + 1 093.3. The value of dv/dP of magnesite is greater than the previous data obtained by Gillet, which was mostly taken under the mantle pressure. And at the ambient temperature, magnesite can be used as a pressure gauge, and the relation between the pressure and Raman shift of 1 094 cm(-1) peak position is given as following: P(MPa) = 125.8 x (deltavp) 1094 + 124.7 (1094 cm(-1) < vp < 1101 cm(-1)). PMID- 16379282 TI - [Structural studies on methanol up to 563 K under pressure]. AB - This paper investigates the structure of methanol by Raman spectrum in diamond anvil cell up to 563 K. The result shows that pressure increases at elevated temperature, but the effect of pressure on the stretching vibrational modes of C H is inverse to that of temperature. The action of temperature is weaker than that of pressure. So the spectral profile of stretching vibrational modes of C-H gradually changes and the Raman shift moves to higher frequency with increasing temperature and pressure, indicating that the bonding energy of C-H bonding increases with temperature and psessure. In addition, the increase of the relative peak area of O-H stretch band with both the temperature and pressure indicates that the total intensity of O-H bonding is greater compared with that of C-H bonding. Thus, it can be inferred that the higher pressure might slow down the transforming process of kerogen to hydrocarbon under the geological condition. PMID- 16379283 TI - [Crystal growth and spectroscopy of Er/Yb:KGW crystal]. AB - The Er3 /Yb3+:KGW crystal with the dimensions of 30 mm x 25 mm x 15 mm was grown from K2W2O7 solvent by modified Czochralski method. The absorption spectrum was measured at room temperature and its absorption peaks were assigned. The emission spectrum was obtained under the excitation of 980 nm. There are two broad emission bands at 1024 and 1535 nm, whose FHWM are 60 and 36 nm respectively. It shows that this crystal is suitable for InGaAs LD pumping, and may be a promising laser crystal tunable at near 1 024 and 1 535 nm. PMID- 16379284 TI - [Spectral study on the interaction mechanism between thionine and calf thymus DNA]. AB - The interaction mechanism of thionine (TH) and calf thymus DNA (CT-DNA) was studied with UV-Vis absorption spectoscopy, fluorescence spectroscopy, circular dichroism (CD), and X-ray photoelectron spectroscopy (XPS). It was demonstrated that in the pH 7.2 phosphate buffer solution, the interaction of TH and CT-DNA is in the intercalative mode. The absorbance of the absorption peak of TH decreases and the peak position shifts to the red direction. The binding constant (K) of TH with CT-DNA is 1.45x 10(4) mol x L(-1). The fluorescence intensities of TH decrease drastically with increasing the concentration of CT-DNA. The Stem-Volmer quenching constant (Ksv) was calculated to be 1.01 x 10(4) mol x L(-1). The intercalative sites are mainly located at the G-C sequences of the CT-DNA molecule through the S atom in the TH molecule according to the experimental data obtained from the fluorescence spectra and XPS analysis. The secondary structure conformation of CT-DNA is changed due to its interaction with TH. PMID- 16379285 TI - [Synthesis and DNA binding spectroscopy studies of Cu(II)-Thr-Phen]. AB - A new complex [Cu (Thr)(Phen)H2O] SO4 . HO2 x CH3 OH (Thr = DL-Threonine, Phen = o-Phenanthroline), which has not been published, was synthesized and characterized by elemental analysis, IR spectroscopy, and TG-DTA. The interaction of the complex and sperm DNA was studied by electronic absorption and ethidium bromide (EB) fluorescence spectroscopy. The result indicates that the maximal absorption peaks of this complex are red-shifted and the intensity is weakened; At the same time, it can to some extent quench the emission intensity of EB-DNA system. Therefore, the authors come to a conclusion that the interaction of this complex and sperm DNA is intercalation. PMID- 16379286 TI - [Interaction of surface active fluorescence probes and bovine serum albumin]. AB - The binding between two surface-active substituted 3H-indole fluorescence probes, i. e., iodo-dihexadecyl methyl-2-(p-dodecyl amino phenyl)-3, 3-dimethyl-5 carboethoxy-3H-indole ammonium and iodo-dimethyloctadecyl-2-(p-dodecyl amino phenyl)-3,3-dimethyl-5-carboethoxy-3H-indole ammonium, and bovine serum albumin (BSA) in aqueous solution was studied using fluorescence. The binding constant and binding site number of molecule 1 and molecule 2 with BSA were obtained. It was confirmed that electrostatic interaction is the primary driving force for the combination of BSA with molecule 1 or molecule 2. According to the Forster resonance energy transfer theory, the distances between molecule 1, molecule 2 and tryptophan of BSA were calculated to be 2.90 nm and 4.02 nm, respectively. PMID- 16379287 TI - [Spectral properties, protonation and fluorescence quantum yield of ciprofloxacin]. AB - Fluorescence spectra, ultraviolet absorption spectra, and protonation of Ciprofloxacin (CIP) at different pH values have been studied. Fluorescence quantum yield of CIP under neutral condition has been measured. In HCl medium with [H+] > 1 mol x L(-1), CIP molecules (simplified as HL) may accept three protons to exist as H4L3+ with very weak fluorescence, and its maximum fluorescence emission wavelength (lambdamax) is 456 nm. In acidic solution of pH 0 to pH 2, CIP mainly exists as H3L2+ form with lambdamax at 450 nm, and fluorescence intensity is relatively weak and increases with increasing of pH. In the range of pH 2 to pH 4, CIP mainly exists as H2L+ form with a strong fluorescence, and lambdamax is still at 450 nm. When pH>4, lambdamax gradually blue-shifts to 414 nm, fluorescence intensity slightly decreases as pH increases, and at the same time an evident change in ultraviolet absorption spectrum is observed, indicating that H2L+ has lost proton to exist as dipole ion form HL. When pH>8, the fluorescence intensity decreases until disappearance as pH increases, indicating that HL has lost proton to exist as non-fluorescence anion ion form L-. In the molecular form changing process, the maximum excitation wavelength of CIP is essentially constant at 275 nm, but the maximum emission wavelength changes obviously. In a buffer solution with pH 7.0, and using quinine bisulphate as a reference, the fluorescence quantum yield of CIP at maximum excitation wavelength 275 nm was measured to be 0.12. PMID- 16379288 TI - [Fluorescence spectroscopy study of humen serum albumin quenched by levofloxacin]. AB - The fluorescence spectroscopy of humen serum albumin (HSA) and the interaction of HSA and the Levofloxacin were studied. When the Levofloxacin was added into HSA solution gradually, an interesting new phenomenon emerged in emission spectrum peak. The intensity of 343.70 nm emission peak of HSA decreased obviously and moved towards long-wavelength, which is atypical quenching phenomenon. It was found that the excitation peak of HSA was positioned at 286.70 nm also. A new emission peak emerged at 503.96 nm, which resulted from the Levofloxacin. The excitation peaks of 503.96 nm fluorescence of Levofloxacin were positioned at 300.16 nm and 336.16 nm. When Levoflaxasin was added into HSA gradually, the 300.16 nm and 336.16 nm excitation peaks moved towards long wavelength. The dissociation constant of Levofloxacin from HSA is about Kd = 3.65 x 10(-5) (mol x L(-1)). The combination constant of Levofloxacin is about Ks = 2.742 x 10(4) (L x mol(-1)). The quenching process of Levofloxacin-HSA is not dynamic quenching, which resulted from the molecular diffusion and collision. It is caused by the static quenching process resulting from the chemical component between molecules. The energy transfer efficiency between Levofloxacin and HSA is E = 0.372. According to these calculation results, the combination position between the binding site of Levofloxacin and the tryptophane of HSA is about R = 1.933 nm. PMID- 16379289 TI - [Fluorescence spectroscopic study on the monoammonium glycyrrhizinate-bovine serum albumin supramolecular system]. AB - The fluorescence quenching mechanism of bovine serum albumin (BSA) by monoammoniumglycyrrhizinate (MAG) has been studied. It is proved that static quenching exists in the MAG-BSA supramolecular complex. The formation constant KA and the thermodynamic functions (such as deltaG, deltaH and deltaS) for the reaction have been all obtained. According to the thermodynamic parameters, the main sort of binding force was electrostatic force. The effect of various metal ions and temperatures on the formation constant of MAG with BSA was also studied. The binding distance between MAG and BSA and the transfer efficiency have been obtained based on the mechanism of Forster energy transfer. The effect of MAG on the conformation of BSA has also been analyzed using synchronous fluorescence spectroscopy. PMID- 16379290 TI - [Study the characteristics of dissolved organic matter and turbidity of water using laser induced fluorescence and laser scattering]. AB - In the experiment a frequency doubled radiation of Nd:YAG laser was used for excitation at 532 nm. The fluorescence spectra of dissolved organic matter (DOM) and chlorophyll a (Chl-a) in several types of water samples with laser-induced fluorescence (LIF) measurements using a 532 nm wavelength excitation source were measured in the laboratory, and the spectral characteristics of DOM and Chl-a were analysed. The characteristics of turbidity were investigated by measuring the amount of scattering light of suspended particles in the water volume. The curve of scattering intensity against corresponding turbidity of SiO2 is showed. The possibilities of water quality monitoring based upon the fluorescence spectral characteristics of contaminations and the turbidity by means of LIF method. PMID- 16379291 TI - [Study on UV-visible DOAS system based on photodiode array (PDA)]. AB - A long-path differential optical absorption spectroscopy (DOAS) system is introduced. A photodiode array is employed as the detector to replace the complicated SD detector which consists of a PMT and a slotted disk. The properties of the detector and the spectrometer unit such as offset, dark current, noise, linearity, resolution, and wavelength range were measured. This system was also tested to measure SO2 and NO2 in the atmosphere. The detection limits of this system for SO2, and NO2 over a 713 m light path were determined. PMID- 16379292 TI - [Determination of fleroxacin by photochemical fluorescence of zinc-fleroxacin complex]. AB - Based on the complex formed by Zn2+ which can strengthen the relative fluorescence intensity of fleroxacin evidently, a novel photochemical fluorescence method was developed. The effects of the acidity, the concentration ratio of Zn2+ to fleroxacin, and the time for illumination were studied. Under the optimum experiment conditions, the linear range of the determination was 5.00 x 10(-8) - 5.00 x 10(-6) mol x L(-1). The detection limit was 4.2 x 10(-8) mol x L(-1). The relative standard deviation of the determination of fleroxacin (5.0 x 10(-7) mol x L(-1)) was 1.7% (n = 20). The method was successfully applied to the determination of fleroxacin in specimens, and the recoveries were in the range of 95.0%-105%. The mechanism of this system is proposed. PMID- 16379293 TI - [Study on the reaction of proteinase from Bacillus subtilis with alizarin complex]. AB - The interaction of bacillus subtilis proteinase(BSP) and Alizarin Complex (ALC) was investigated by spectrophotometric method in acidic solution(pH 4.20). When BSP was added into ALC, a red color was observed, which indicated the formation of BSP-ALC associated compound. The maximum absorption of the red color coordination compound was obtained at 510 nm. It was found that the red shift of maximum absorption of the complex was 85 nm. According to balance percolation method, molar ratio method and double wavelengh method, the apparent molar absorptivity epsilonB = 6.68 x 10(3) L x mol(-1) x cm(-1). Conditional constants were defined, the maximum binding number n = 10, and the apparent binding constant K = 7. 25 x 10(6) L x mol(-1). It was found that Scatchard model is appropriate in the treatment of data obtained here. PMID- 16379294 TI - [Spectral research on the effect various metal ions on the of photocatalytic degradation properties of TiO2 nanocrystal]. AB - TiO2 nanocrystalline was prepared by a emulsion method in the mixed solvent of cyclohexane and acetic anhydride using, titanyl organic compound as the precursor. The prepared product was characterized by means of X-ray diffraction (XRD) and Fourier transform infrared spectrum (FTIR). The photocatalytic oxidation of methyl orange with TiO2 loaded with various metal ions was discussed. The spectral research results showed that the photocatalytic reaction of methyl orange with TiO2 loaded with Bi3+ was promoted, some metal ions caused almost no promotion of TiO2 oxidation of methyl orange, and Ni2+ and Fe3+ ions caused a little promotion. The best mass proportion of Bi3+ and TiO2 was defined as 1:8, which has a good photocatalytic activity. The photocatalytic degradation reaction of methyl orange was affected by the concentration of TiO2 nanocrystal, and when the mass proportion of Bi3+ and TiO2 is 1:8, the best photocatalytic activity concentration of TiO2 is about 1.6 g x L(-1). PMID- 16379295 TI - [Spatial orientation interaction mechanism of three component complex of protein- BSA/SDS/Azur a system]. AB - The interaction of bovine serum albumin (BSA), sodium dodecyl sulphate (SDS) and Azur A(AA) was studied by spectral probe. The influence of the molar ratio of SDS/BSA on the absorption spectra of BSA-SDS-AA complex was investigated. It was found that the new absorption peak and color changes generated by BSA-SDS-AA complex arose from the difference in the spatial orientation aggregative degree of AA binding on BSA-SDS-AA complex. PMID- 16379296 TI - [Synthesis and spectral characterization of rare earth complexes of acylpyrazolone-beta-alanine]. AB - In non-aqueous solvent, a new amino-acid schiff base, 1-phenyl-3-methyl-4-benzoyl pyrazolone-5-beta-alanine (HL) was synthesized by a reaction of beta-alanine with benzoylpyrazolone, and its ten rare earth complexes were obtained from refluxing a solution of schiff base and rare earth nitrates. On the basis of elemental analysis and molar conductance, the general formula of the complexes, [REL2NO3] nH2O (RE=La, Sm, Eu, Tb, Y, n=2; RE=Pr, Nd, n = 1; RE=Dy, Er, Yb, n=3), is given. These were characterized by IR, UV-Visible, 1H NMR, 13C NMR and fluorescence. The results show that the schiff base is a tridentate ligand, and the rare earth ions exhibit a coordination of eight in the complexes. In visible spectra, the supersensitive transitions of the Er complex at 522 nm (4 I15/2 -->2 H11/2, 4 S3/2 ) and that of the Nd complex at 573, 584 nm (4I9/2 -->2 G7/2 + 4G5/2) can be observed. Fluorescence of the complexes was produced principally by f-f transition of central ion RE3+, and the ligand has little influence on lightening role. The order of relative intensity of fluorescence of the complexes is ITb > ISm > IEu > IDy. PMID- 16379297 TI - [Thermochemical studies on the binding reaction of perseleno diphenyl 2,2' diformic acid with bovine serum albumin]. AB - The binding of newly compounded perseleno diphenyl 2,2'-diformic acid to bovine serum albumin(BSA) was studied at different temperatures using fluorescence spectrum and UV spectrum. The fluorescence quenching data was analyzed according to Stern-Volmer equation and Lineweaver-Burk double-reciprocal equation. It was showed that this quenching complies better with the charactristic of static fluorescence quenching. The binding constant, thermodynamic parameters, and the binding spot of the compound with certain structure, coming from perseleno diphenyl 2,2'-diformic acid and bovine serum albumin, were obtained. Besides, themechanism of static fluorescence quenching and the quality of binding power were both discussed. The information of the binding mode, the mechanism of its transportation, and some medical theories in human body were offered. PMID- 16379298 TI - [The reaction mechanism between cefoperazone and human serum albumin]. AB - The reaction mechanism between cefoperazone and human serum albumin (HSA) and the affinity between cefoperazone and beta-lactamase were investigated by spectrometry and spectrofluorimetry. The interaction dissociation constants of human serum albumin and cefoperazone have been determined from a double reciprocal Lineweaver-Burk plot. The binding distance and the transfer efficiency between cefoperazone and HSA were also obtained according to the theory of Forster' non-radiation energy transfer. The result suggested that the main binding force between cefoperazone and HSA is electrostatic force interaction. The high beta-lactamase stability of cefoperazone may be correlative with its molecular structure. The antibiotic activity and valency connect with transfer efficiency and dissociation constant. The effect of cefoperazone on the conformation of HSA was also analyzed using synchronous fluorescence spectrometry. PMID- 16379299 TI - [Spectrum studies and application of the reaction system phenanthroline-EBT H2O2]. AB - A kinetic spectrophotometric method for the determination of trace phenanthroline was proposed. The method is based on the inhibition of phenanthroline to the discoloring reaction of EBT oxidized by hydrogen peroxide in the dilute acetic acid solution. At the same time, the kinetic parameters were studied in detail. Under optimum experimental conditions, the linear range is 0.003-0.08 mg x L(-1), and the detection limit is 4.0 microg x L(-1). The method is simple, rapid, and easy to operate. The method was applied to the determination of trace phenanthroline in water samples with satisfactory results. PMID- 16379300 TI - [Simultaneous determination of germanium and some major ash forming elements in lignites using a dual-view ICP-OES]. AB - In this article, an ICP-OES method of simultaneous determination germanium and some major ash forming elements especially for Fe, Al, Ca, Mg, K, Na, Ti in lignites is described. An ICP-OES instrument equipped with a dual-view plasma torch and a simultaneous detector was used. This technology allows the simultaneous determination of trace elements and major components in the axial view along with in the radial view. The moderate operation condition of the instrument and analytical wavelength lack of interference from elements in the sample matrix was selected, the spectral background was automaticly eliminated. The obtained coal ashes were dissolved in mixtures of nitric acid, hydrofluoric acid and perchloric acid after coal samples were cinerated. The analytical results were in good agreement with those provided by national standard methods. The detection limit of the instrument is ranged between 0.00039-0.10 microg. x mL(-1), the precision of the method is in the range of 0.79%-2.84%, the average recovery of samples is 92.38%, the proposed method is well suitable for the determination germanium and some major ash forming elements in lignites. PMID- 16379301 TI - [Determination of ten trace rare earth elements in the sample by atomic emission spectrometry]. AB - This paper describes the determination of trace La, Ce, Pr, Nd, Sm, Gd, Tb, Dy, Yb and Lu in the sample, using carbon powder, potassium sulfate, barium sulfate, strontium sulfate, and scandium chloride as buffer, by atomic emission spectrometry (AES). Scandium was selected as internal standard line. Sample separation and chemical treatment were not required. The sample was directly loaded into ordinary electrode. The method is simple, rapid and accurate. The determination requirement and influence factors were studied. A new method was developed for the determination of ten rare earth elements, for which the detection limit is smaller than 0.030%, and the range of the recovery is 94% 105%. The results of these elements in standard sample are in agreement with certified values, and the RSD is smaller than 5% (n = 9). The method has been applied to the determination of ten rare earth elements with satisfactory results. PMID- 16379302 TI - [Determination of cadmium, chromium and lead in Lycoris radiata with microwave digestion technique by graphite furnace atomic absorption spectrometry]. AB - A new method using microwave digestion technique was developed for the determination of cadmium, chromium and lead in Lycoris radiata by graphite furnace atomic absorption spectrometry (GFAAS). Digestion solvents were discussed for sample preparation by microwave digestion technique. The optimum condition of determination by GFAAS was studied in the presence of NH4H2PO4 matrix modifier. The detection limits of Cd, Cr and Pb are 0.066 7, 1.22 and 0.810 microg x L(-1), respectively. The method was applied to the determination of Cd, Cr and Pb in Lycoris radiata samples with satisfactory results. The RSD of determination were lower than 3.1%. The recoveries were from 83.8% to 118%. PMID- 16379303 TI - [Determination of copper, zinc, iron, calcium and magnesium in Pueraria lobata ohwi by FAAS]. AB - The samples of Pueraria lobata ohwi were digested by HNO3 + HClO4. Copper, zinc, iron, calcium and magnesium in pueraria lobata ohwi from Da Bieshan were successfully determined by flame atomic absorption spectrophotometry (FAAS). The results showed that there are rich grand elements, such as Ca and Mg, and profitable elements, such as trace elements Cu, Zn and Fe, in Pueraria lobata ohwi. The content sequence of metal elements is as follows: Copper: leaves > flowers > rattans > roots; Zinc: leaves > flowers > rattans > roots; Iron: roots > rattans > leaves > flowers; Calcium: leaves > roots > flowers > rattans; Magnesium: flowers > roots > leaves > rattans. The results provided useful data for discussing the relationship between the metal elements in Pueraria lobata and hypertension diseases. ohwi and the cure for cerebrovascular diseases. PMID- 16379304 TI - [Determination of fourteen trace elements in chinese traditional medicines by atomic absorption spectrometry]. AB - Seven kinds of Chinese traditional medicines, including laoniankechuan tablet, fufangbanxia tablet, weitongning tablet, quanshen tablet, shengshijiangtang capsule, xiasangju particle, and American yangshen tablet, were digested with HNO3-HClO4 mixed acid. The fourteen trace elements, including calcium, magnesium, iron, zinc, potassium, sodium, manganese, copper, chromium, cobalt, strontium, nickel, cadmium and lead in the drugs were determined by atomic absorption spectrometry. The effects of the type of mixed acid, the ratio of HNO3 to HClO4 in mixed acid, the volume of digesting solution, and the digesting time were also investigated in detail. The results obtained show that the concentrations of Ca, Mg, Fe, K and Na in seven kinds of Chinese traditional medicines are higher than those of other elements. Moreover, shengshijiangtang capsule for the treatment of diabetes contains plenty of Mn, and weitongning tablet for the treatment of stomach disease contains plenty of Sr, Mn and Cu. PMID- 16379305 TI - [Preconcentration with crosslinked chitosan for determination of trace manganese in waters by flame atomic absorption spectrometry]. AB - A novel method for the preconcentration with crosslinked chitosan (CCTS) and determination of trace manganese in waters by flame atomic absorption spectrometry has been presented. The absorption rate of manganese (VII) by CCTS was 98% at pH 3.00. The effect of preconcentration time, dosage of CCTS, temperature, sample volume and co-existing elements have been investigated. The mechanism of adsorption of CCTS for Mn(VII) was also discussed. The detection limit of the method was 1.86 microg x L(-1), and the relative standard deviation (RSD) was 2.0% (n = 10). The method has been applied to the determination of trace manganese in south lake and Changjiang water with satisfactory results. PMID- 16379306 TI - [Analysis of lead contents in edible nuts by AAS]. AB - The contents of the pollutant element, lead, in edible nuts were determined by flame atomic absorption spectrophotometry. The linear relationship over the range of 0.00-2.00 microg x mL(-1) is good (r = 0.9991). The recovery is in the range of 94.0% - 102.0% and the RSD is below 4.64% (n = 5). The method is simple and the results are accurate. It can be used as a monitor method for lead in edible nuts. The results show that the pollutant element lead exists commonly in edible nuts. The plants were affected by water, soil and atomsphere during growing, and can absorb lead in environment. PMID- 16379307 TI - [The effect of the photoelectric detector on the accuracy of the spectrometer]. AB - An optimized photoelectric detector will increase the precision of a spectrometer, thus indicates an important way to develop high performance spectrometer. With an eye to this, a model describing the process that spectrogram is integrated and sampled by photoelectric detector and restored after low-pass filtering is developed. Based on the model, the influence of the characteristic parameters of the detector on the spectral line in the frequency domain is analyzed and the relation between the full width half maximum (FWHM) of the spectra line and the integral interval, sampling space and sensitivity of the detector is deduced. The conclusion indicates that both the integral interval and sampling space should be 1/6 of the FWHM for a spectral line with gaussian profile as a result of compromise between accuracy and feasibility. Moreover, the critical point deciding the right situation for scanner and array detector is given. Other guide line to optimize the photoelectric detector and increase accuracy is suggested also. PMID- 16379308 TI - [Exact expressions for bend phenomena of spectral line and spectral band of spectrometer]. AB - In this paper the grating equation and integrate expression of diffractive polar angle and the azimuthal angle with arbitrary on the vector diffraction theory. The exact calculation formulae for bend quantity of spectral line angle of incidence are described based and spectral band of plane grating spectrometer are given, and two kinds of angular dispersion expression of correspondingdiffractive polar angle and azimuthal angle are also presented. The calculated results of the exact spectral line bend formula are compared with those of approximate formulae. The effects of the height of incident slit on dispersion capability are examined by values. Because no approximations are performed during the above deduction of each formula and all factors that lead to the bend of spectral line and spectral band are concerned, the given formulae in the paper are more comprehensive and reliable than all the approximate formulae used before. Consequently, these formulae can be used as a theory criterion by which the actual spectral instrument is designed, tested, equipped, adjusted and used. PMID- 16379309 TI - [Detection of trace organic compounds by using ion mobility spectrometry and SIMPLISMA]. AB - Ion mobility spectrometry (IMS) is a sensitive means for detection of trace volatile organic compounds. This technique has been used as a routine method for some organic compounds in forensic science, environmental and pharmaceutical analysis, etc. SIMPLISMA (simple-to-use-interactive self-modeling mixture analysis) is an effective method for large-scale data processing. In the present work, SIMPLISMA has been applied to process IMS data of three methanephosphonate esters. The SIMPLISMA can retain the peak characteristics and eliminate much of the noises of the IMS measurements. The processed spectral data can be subsequently used for other computations, such as curve resolution and pattern recognition etc. PMID- 16379310 TI - [Determination of mean electron energy in O2 non-equilibrium plasma by relative spectrum line intensity]. AB - The process of O2 non-equilibrium plasma is simulated by Monte Carlo method. The electron drift velocity as a function of E/N is given, which is in good agreement with experimental result. The variations of mean electron energy and the number density of electronic excitation states of O2 with the reduced field E/N are simulated. A method to determine the mean electron energy by relative spectrum line intensity is proposed, which is of great importance to the applications of non-equilibrium plasma in many fields such as material preparation, laser application, micro-electronics technology and so on. PMID- 16379311 TI - [Encephalopathy and encephalitis--influenza-associated neurological sequels]. AB - We would like to present chosen neurological sequels post viral infections- viruses of influenza A and B. Unfortunately pathogenesis of encephalopathy and encephalitis after infection with viruses of influenza is still unclear. This problem seems very important because a most of neurological cases--encephalopathy and encephalitis after influenza concerns children and adolescents, who haven't yet full immunological protecting mechanisms. The basic manner of protection against influenza and all post influenza sequels is immunization, repeated every epidemiological season. This method is the most effective against viruses of flu. PMID- 16379312 TI - [Analysis of intracellular cytokines IFN-gamma and IL-4 in peripheral blood T cells in children with bronchospastic reaction]. AB - In the epidemiological study there was suggested that respiratory tract infections--were a strong risk factors for the beginning and development of bronchial asthma. The aim of the study was the evaluation of intracellular cytokine IL-4 and IFN-gamma on peripheral blood T subsets in children with atopic asthma (AA) and recurrent respiratory tract infection with bronchospasm (RRTI). METHOD: Peripheral blood T cells were stained with fluorescence-labelled antibodies specific for intracellular cytokines IFN-gamma and IL-4 and cell surface markers CD3, CD4 and CD8, and were subjected to flow-cytometric analysis. RESULTS: Comparing peripheral blood lymphocytes of atopic asthma patients with those of recurrent infections we found significantly more cells positive for IL-4 in asthma patients than in recurrent infection patients, both in the CD3+ subsets (p<0.03) and CD4+ subset (p<0.01). We have also found that percentage of CD4+ was significantly lower (p<0.007) and percentage of CD8+ cells was significantly higher (p<0.05) in RRTI group comparing to atopic asthma patients. In AA group there was a significant increase intracellular expression of IL-4 among the CD3+ (p<0.03) and CD4+ (p<0.01) subsets and no significant differences among CD8+ subset. In AA group there was a significant decrease ratio of IFN-gamma/IL-4 among all of the evaluated subsets. CONCLUSION: Basing oneself on these results we conclude that markers of atopy are: increased intracellular expression of IL-4 among CD4+ cells and decreased IFN-gamma. PMID- 16379313 TI - [Lipid peroxidation product as prognostic factors in acute necrotizing pancreatitis]. AB - The aim of the study is the assessment of the role of oxygen free radicals (OFRs) in the pathology of acute pancreatitis (AP). MATERIAL AND METHODS: It has been accomplished based on the analysis of lipid peroxidation products' and hydrogen peroxide blood concentration in 60 patients with AP. Spectrometric and spectrofluorometric methods were used. Patients were divided into 2 groups. Group 1 consisted of 42 patients, who survived the 30 day observation period. Group 2 included patients, with a systematic deterioration resulting in death. All patients were subjected to surgery between the 15th and 30th day of observation because of septic syndrome signs. On 1st, 5th, 10th, and 15th day of treatment we measured concentrations of lipid peroxidation products such as: conjugated dienes (CD), malonyl dialdehyde (MDA), Schiff's bases, lipid peroxides (LOOH), and hydrogen peroxide. RESULTS: We observed high levels of CDs and MDA throughout the entire observation period as well as increase in lipid and hydrogen peroxide serum concentrations in patients with poor prognoses. Shiff's bases serum concentrations presented a non-characteristic pattern of changes. CONCLUSIONS: Lipid peroxidation products' metabolism path observed in our study suggests a substantial role of oxygen free radicals in acute pancreatitis pathology. Authors claim, that the assessment of lipid peroxidation product serum concentrations may be a useful marker for prognosing in most severe forms of acute pancreatitis. PMID- 16379314 TI - [Serum adiponectin levels in patients with acromegaly]. AB - Acromegaly is frequently associated with impaired glucose tolerance or diabetes. Insulin resistance has been reported to be related to low levels of adiponectin, which is exclusively produced by adipose tissue, as well as to decreased concentration of sex hormone binding globuline (SHBG). Growth hormone (GH) excess is associated with increased muscle mass and decreased fat mass. Influence of GH on production and secretion of adiponectin is not completely understood. The aims of this study were to assess serum adiponectin levels in patients with active acromegaly and to compare them with concentrations in the control group and to investigate relationships between adiponectin and indexes: body mass index (BMI) and waist-to-hip ratio (WHR) as well as SHBG levels. MATERIALS AND METHODS: Serum adiponectin and SHBG levels were measured by radioimmunoassay in 30 patients with active acromegaly and in 22 sex-, age-, BMI- and WHR-matched controls. The groups were comparable in prevalence of hypertension and disturbances of glucose metabolism. RESULTS: Serum adiponectin concentrations were higher in acromegalic patients compared with control group (24.03 +/- 9.85 mg/ml vs. 14.46 +/- 3.96 mg/ml, p<0.001). When the subjects were fallen into two groups according to sex, significant differences were also observed. Serum SHBG levels were lower in cases compared with controls (25.36 +/- 22.89 nmol/l vs. 33.54 +/- 15.07 nmol/l, p=0.03). In the control group we found adiponectin positively correlated with SHBG concentrations (r=0.53, p=0.01) and negatively correlated with BMI (r=-0.52, p=0.02) and WHR (r=-0.51, p=0.03). However, these relationships were not observed in patients with acromegaly. No significant associations were found between adiponectin and GH or IGF-I. CONCLUSIONS: Serum concentration of adiponectin is significantly higher in patients with acromegaly when compared with the control group and is independent of BMI, WHR and SHBG. The results suggest that adiponectinemia does not directly affect development of insulin resistance in acromegaly. PMID- 16379315 TI - [Diagnostic value of tissue polypeptide antigen in serum of larynx cancer patients]. AB - Tissue polypeptide antigen (TPA) is a serological tumour marker used in the diagnosis, management and follow-up of patients with head and neck cancer. The aim of this study was to evaluate the diagnostic and prognostic significance of pretreatment TPA serum levels in patients with larynx cancer. The predicting ability of this tumour marker with respect to histological type, pathological state and lymph node metastasis was also assessed. MATERIALS AND METHOD: Concentrations of the TPA in the serum from 35 patients were measured by immunoradiometric assay. RESULTS: The results showed the sensitivity value for the group of 35 patients was 29.4%, but in clinical stage IV was 70%. TPA levels correlate with T classification and lymph node metastasis. In T4 tumors it was significantly higher than in T2 (p=0.047). TPA levels were significantly higher in patients with nodal invasion and were generally lower in patients with spinocellular carcinoma (p=0.0048). CONCLUSION: Our date indicate that TPA is of limited usefulness in the primary diagnosis in patients with larynx cancer, but is useful in detecting lymph node metastasis. PMID- 16379316 TI - [Antioxidant barrier in patients with gastric cancer--preliminary study]. AB - Gastric carcinoma is one of the most common malignancies of causes of cancer death world-wide, although its incidence and mortality and decreasing, particularly in developed countries. One of the many factors in its etiopathogenesis are free oxygen radicals and their derivatives, excess of which comes from disturbances of enzymatic and nonenzymatic mechanisms of antioxidant barrier. The aim of this study was the evaluation activities of antioxidant enzymes: superoxide dismutase (CuZnSOD, MnSOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione transferase (GST), glutathione reductase (GSH-R) as well as measurement of glutathione (GSH) and malondialdehyde (MDA) levels both in tissues (cancer and healthy) and in blood serum obtained from patients with gastric cancer. RESULTS: MnSOD and glutathione dependent enzymes (GSH-Px, GST, GSH-R) activities in gastric cancer were found to be higher than those in healthy stomach tissues. In pre- and postoperative blood serum from patients with gastric cancer was observed increase of CAT and GST as well as decrease of CuZnSOD and GSH-Px compared with the blood serum from healthy blood donors (control group). In preoperative blood serum from patients with gastric cancer GSH-R activity was lower than in control serum, whereas in postoperative serum it was higher compared with serum of the control group. Reduced GSH level was found to be lower in gastric cancer than this in healthy stomach tissue, adjacent to the cancer. Its level was also lower in pre- and postoperative blood serum of patients with gastric cancer compared with level in control serum. MDA concentration was found to be much higher in gastric cancer than this in healthy stomach tissue, whereas weren't significant differences between level of MDA in pre- and postoperative blood serum. CONCLUSION: Our results showed that functioning of antioxidant barrier of organism is severely impaired in patients with gastric cancer. PMID- 16379317 TI - [Assessment of 8-hydroxy-2'-deoxyguanosine concentrations in bladder cancer patients treated with intravesical BCG instillation]. AB - Increased 8-hydroxy-2'-deoxyguanosine (8-OHdG) concentration has been proposed as reliable marker of the oxidative DNA damage, and prognostication of urological carcinogenesis, particularly in bladder cancer. Widely accepted method of treatment in early stages of bladder cancer is transurethral electro resection (TURN). Intravesical bacillus Calmette-Guerin instillation is considered to be the therapeutic agent for superficial transitional cell carcinoma (TCC) of the bladder and has been established as standard therapy in the patients. The aim of the study was to test the concentration of 8-OHdG in urine in patients with bladder cancer and the effect of TURN and BCG therapy on the 8-OHdG value. MATERIAL AND METHODS: We tested 12 patients (3 female and 9 male) with superficial bladder cancer and 31 healthy controls. Urine for the examination was drawn in 4 time points: before and 2 weeks after TURN treatment, then 6 weeks after 6 intravesical instillations of BCG (Onko BCG--BIO MED Lublin) by Morales and 12 weeks after TURN. 8-OHdG concentration in urine was tested using ELISA commercial kit (OXIS health) and the values of 8-OHdG are expressed as ng/ml of urine. RESULTS: Patients with superficial bladder cancer had 16.89 ng/ml of 8 OHdG in urine before the TURN procedure. The value was significantly (p< 0.005) higher then 12.98 ng/ml noted in healthy controls. 2 weeks after the procedure the 8-OHdG level decreased to 13.36 ng/ml. After 6 weeks of repeated 6 intravesical instillations of the BCG the concentration of 8-OHdG dropped to 10.91 ng/ml (p<0.005) and returned to the value 13.28 ng/ml after the next 4 weeks. CONCLUSIONS: Patients with bladder cancer have significantly increased concentration of 8-OHdG in urine compared to controls. 8 week combine therapy with TURN and BCG resulted in a significant decrease in 8-OHdG concentration in urine The beneficial effect of BCG instillations seems to result from strengthening of the antioxidative DNA protection. PMID- 16379318 TI - [Subjective symptoms related to mobile phone use--a pilot study]. AB - Research findings indicate that the use of mobile phones may lead to a number of symptoms such as headache, impaired concentration and memory, fatigue. In Poland this problem has not as yet been addressed by scientific studies. THE AIM: The present project was undertaken to investigate whether the symptoms of ill health reported by young people may be associated with the use of mobile phone. MATERIAL AND METHODS: A survey using a self-reported questionnaire was conducted among randomly selected university students in Lodz, Central Poland. The questionnaire was designed specifically for this study and contained items on health condition and complaints as well as on frequency of mobile phone use. The number of questionnaires necessary for the study was assessed using the simple random sample method. Out of the 160 copies distributed among the students, 140 (87.5%) were completed. Eventually, 117 questionnaires were subject to analysis; the data from respondents who reported health problems (neck trauma in a car accident, chronic sinusitis and arterial hypertension) were excluded. The following statistical methods were used to analyse questionnaire data: t-Student test for equal and unequal variances or F-Snedecor test for comparing parameters in two study groups, Fisher exact test for comparing frequency, and single and multiple logistic regression models for quantitative risk assessment of negative health outcomes in relation to exposure level and with control for confounders. The subjects were 61 (52.1%) males and 56 females (47.9%). RESULTS: Most of the subjects (62%) assessed their health condition as good, 31% as very good and 7% as fair. 70% complained of headache and 20% of dizziness. Impaired concentration occurred in 56% of respondents. Facial dermatitis was reported by 11%. The most prevalent symptom related to mobile phone use was the thermal sensation within the auricle and behind/around the ear. This was reported by 33 subjects (28.2%). Out of 82 subjects who complained of headache, only 8 (6.8%) related this symptom to mobile phone use. Only 10 subjects of 65 reporting impaired concentration thought it could be associated with their using a mobile phone. The symptoms and health complaints reported by the respondents in no case were the reason for a medical check-up or taking any medication. CONCLUSIONS: The large number of young people complaining of headache and impaired concentration calls for further research to investigate the underlying reasons. It cannot be excluded that one of them may be exposure to EMF emitted by mobile phone. The explanation should be sought through further experimental and epidemiologic studies. PMID- 16379319 TI - [Assessment of oral cavity mucosa and teeth state in patients with chronic obstructive pulmonary disease--part I]. AB - The aim of the study was to assess the state of oral cavity mucosa and teeth in patients with chronic obturative pulmonary disease (COPD). MATERIAL AND METHODS: The study comprised 100 patients with COPD (68 men and 32 women), aged 30-83 years (mean 63.1 +/- 10.17 years). All patients demonstrated II grade COPD (moderate COPD). FEV1/FVC<70% and FEV1 was within 79% and 50% of the predicted value. Beta2-adrenomimetic inhalants or in nebulization, theophylline were applied in the therapy. The control group consisted of 101 subjects (38 men and 63 women), aged 42-88 years (mean 65.3-10.36 years), with no respiratory tract ailments. The rate of smoking was similar in both groups. The examination of oral cavity mucosa and teeth state was performed in artificial lightening with the use of probe and mouth mirror. The number of lacking teeth and the advancement of caries were determined. RESULTS: Thrush appeared to be the most frequent oral cavity mucosa ailment in COPD patients. Toothlessness and reduced teeth number were observed significantly more often in COPD patients than in the control group (p<0.05). The frequency and the type of caries did not differ significantly between the two groups (p>0.05). PMID- 16379320 TI - [Assessment of periodontal state in patients with chronic obstructive pulmonary disease--part II]. AB - The aim of the study was to estimate the periodontal state in chronic obstructive pulmonary disease (COPD). MATERIAL AND METHODS: The study comprised 100 patients with COPD (68 men and 32 women), aged 30-83 years (mean 63.1 +/- 10.17 years). All patients were in the II grade COPD (moderate COPD). FEV1/FVC<70% and FEV1 was within 79% and 50% of the predicted value. Beta2-adrenomimetic inhalants or in nebulization, theophylline were applied in the therapy. The control group consisted of 101 subjects (38 men and 63 women), aged 42-88 years (mean 65.3 10.36 years), with no respiratory tract diseases. The rate of smoking was similar in both groups. To examine periodontal state, periodontal pocket depth was measured (with periodontal sound), Periodontal Index (PI) and dental plaque index were assessed. RESULTS: The frequency and severity of periodontal diseases in COPD patients were demonstrated to be significantly increased as compared to the control group (p<0.05). Adult chronic periodontitis (ACP) was observed significantly more frequently in COPD patients (84.21%) than in the control group (59.74%, p<0.05). A positive correlation was found between periodontal pocket depth and dental plaque index in patients with COPD. CONCLUSION: The frequency and intensification of periodontal diseases speak for their cause-and-effect relationship with COPD. The prophylaxis and treatment of periodontal diseases should be of great importance in the COPD therapy, which may increase its effectiveness. PMID- 16379321 TI - [The coexistence of atopic dermatitis and psoriasis in a 12 months-old girl]. AB - The coexistence of atopic dermatitis and psoriasis is especially rare diagnosed disease in small children. Authors present a 12 months-old girl with both of these diseases. It is important to underline that early diagnosis can apply proper coexistence therapy. PMID- 16379322 TI - [Recurrence of ovarian cancer in extraperitoneal localization after 29th years of radical surgery]. AB - The study presents a case of ovarian cancer recurrence in extraperitoneal localization observed after twenty nine years of radical surgery and radiotherapy. New focus of ovarian cancer has been placed near right oval ligament of uterus. Long-term survival and ovarian cancer recurrence in extra abdominal place may be taken as a confirmation an opinion about a role of multifocal neoplasia of mesothelium in pathogenesis and recurrence of ovarian cancer. PMID- 16379323 TI - [Application of heart rate variability in prognosis of patients with diabetes mellitus]. AB - The ECG based heart rate variability (HRV) measurements has become an important method of assessment of the cardiac autonomic regulation. There is a large body of clinical and experimental evidences indicating that reduced HRV is a predictor of death in patients after myocardial infarction and with heart failure. Diabetic autonomic neuropathy (DAN) is a serious and common complication of diabetes. Cardiovascular autonomic neuropathy (CAN) is the most studied and clinically important form of DAN. Published data demonstrate that CAN is strongly associated with an increased risk of myocardial ischaemia, serious arrhythmias, exercise intolerance and mortality. The determination of the presence of CAN is usually based on Ewing's tests. Measurement of HRV is readily available method and serves to establish diagnosis of autonomic dysfunction. The reduction in parameters of HRV seems not only to carry negative prognostic value in patients with diabetes but also to precede the clinical expression of autonomic neuropathy HRV evaluation in diabetes can encourage physician to improve further therapy. PMID- 16379324 TI - [Remodeling of the basement membrane and airway epithelium in patients with asthma]. AB - Chronic inflammation is recognized as a primary factor in the development of asthma, and its role in the induction of the remodeling of the airways has recently been emphasized. Changes in airway structure such as thickening of the airway wall are common findings in cases of asthma. Thickening of the epithelial reticular basement membrane has been reported to show a positive correlation with airway hyperresponsiveness, the frequency of asthma attacks, and the numbers of fibroblasts and myofibroblasts that lie external and adjacent to it. Studies on the pathology of asthma showed changes of airway epithelium. The most common of these changes are squamous cell metaplasia, goblet cell hyperplasia and damage and shedding of airway surface epithelium. PMID- 16379325 TI - [Omapatrilat--new drug for patients with hypertension and heart failure]. AB - Omapatrilat is a dual angiotensin converting enzyme and neutral endopeptidase inhibitor. It is another a new class of drugs, which are effective in the treatment of patients with hypertension and heart failure also accompanying diabetes. They are producing difference endocrine changes, vasodilatation, diuresis, natriuresis and they have anti hypertrophic activity on tissues, reduce sympathetic tone. Omapatrilat produces beneficial effects in experimental animals and in clinical studies. The overall safety with omapatrilat appears to be good. Angioedema is a rare but potentially life threatening side-effects of endopeptidase inhibitors. Treatment with omapatrilat has a higher tendency towards preventing death and worsening heart failure. PMID- 16379326 TI - [The role of CD44 in pathogenesis of atherosclerosis]. AB - The pathogenesis of atherosclerosis, which remains the most important reason of mortality, is not well understood. Its present conception includes recruitment of inflammatory cells to the vessel wall, activation of vascular smooth muscle cells and endothelium. Much evidence indicate that CD44 molecule, the main receptor for hyaluronan, takes part in pathogenesis via several mechanisms. It mediates in lymphocyte T and monocyte adhesion to the endothelium, stimulates proinflammatory cytokine release from macrophages and participates in dedifferentiation phenotype of smooth muscle cells from contractile state to synthetic one. Furthermore CD44 acts in atherosclerotic plaque destabilization and neointimal proliferation after percutaneous vascular interventions. Numerous studies have supported the concept that CD44 participates in many stages of atherosclerosis. These studies suggest that inhibition of CD44 function may provide an effective mean for inhibiting atherosclerosis, but we still need further investigations for possible therapeutic implications. PMID- 16379327 TI - [Apoptosis mononuclear cells in sarcoidosis]. AB - Sarcoidosis (SA) is a systemic granulomatous disorder of unknown etiology. The diagnosis is firmly established when clinicoradiological findings are supported by histological evidence of noncaseating epithelioid cell granulomas found on tissue biopsy Diagnosis of SA requires exclusion of other causes of granuloma formation, first of all Mycobacterium tuberculosis. Sarcoidosis is a disease known to be associated with various immunological alterations, including depression of cutaneous delayed-type hypersensitivity and an activation of monocytes/macrophages and a heightened helper T cell type 1 immune response at sites of disease but the mechanism leading to the persistent accumulation of inflammatory cells is not understood. In some studies, apoptosis is considered as factor in pathogenesis of sarcoid and tuberculous granuloma. Apoptosis- programmed cell death is a genetically encoded cell elimination program which ensures equilibrium between cell proliferation and cell death and by which crippled, useless or infected cells are eliminated as part of normal physiology. Aberrations in signalling, in receptors or alterations in mechanism of apoptosis are involved in the occurrence of tuberculosis. To date, there are a few of papers concerning alternations of apoptosis of peripheral and alveolar mononuclear cells in sarcoidosis have been published but the results have been controversial. PMID- 16379328 TI - [How to manage the patients with hypoxic pulmonary hypertension?]. AB - Hypoxic pulmonary hypertension (HPH) develops in many patients with advanced chronic obstructive pulmonary disease (COPD) and aggravates their quality of life and prognosis. The most proper but as yet not satisfactory management of HPH involves COPD prophylaxis. In this article pathophysiological background to use systemic vasodilators in treatment of HPH are described, and hemodynamical and clinical results of short and long term trials are discussed. Authors explain lack of clinical benefits with currently used systematic vasodilators in HPH and indicate promising preliminary results with 5-phosphodiesterase inhibitors. The substantial role of chronic oxygen therapy and pulmonary rehabilitation in management of HPH is emphasized. PMID- 16379329 TI - [Free-radical reactions in diabetes mellitus]. AB - Type 2 diabetes mellitus (DM), characterized by decreased insulin secretion, hypoinsulinemia and/or insulin resistance, accounts for 90% of all cases of DM. Oxidative stress is considered as an essential prerequisite for the pathogenesis of this disease. On one hand, current theory of oxidative stress is associated with autooxidation of glucose, which leads to the reactive ketoaldehydes formation, on the other hand it intensifies non-enzymatic glycation of proteins. The increase in reactive oxygen species (ROS) production and a decline in the activity of antioxidants is not only a result of hyperglycemia, but also of hyperinsulinemia and decreased tissue sensitivity to insulin. Because of a significant role of (ROS) in the pathogenesis of chronic diabetic complications, efforts should be made to diminish their toxic effects. Scientific research revealed that the best method for the prevention and treatment of diabetic complications was a long-term compensation of diabetes. To obtain optimal glycemia is crucial, since hyperglycemia is the main source of oxidative stress in patients with diabetes mellitus. PMID- 16379330 TI - [Personal insulin pumps--from experiment research till recommended method of diabetes treatment]. AB - Diabetes treatment with continuous insulin infusion with the use of personal insulin pumps secures strict metabolic control in diabetes--patients and can minimize a number of complications of diabetes. According to American Diabetes Association (2001), diabetes therapy with continuous insulin infusion is recommended for each person, who accept this therapy method. Personal insulin pump is well tolerated device, which enables to conduct a comfortable continuous insulin infusion. The patients, who comply with medical instruction and perform self-monitoring are fit for diabetes treatment with personal insulin pump. According to present recommendation, there is not necessary to hospitalize patients to initiate continuous insulin infusion. Treatment with personal insulin pump takes up in Poland only in wards of Diabetology and Diabetes outpatient clinics. The main indications of personal insulin pump use are: unstable course of diabetes despite of the another intensive insulin methods used before, often hypoglycemic episodes in anamnesis, active, changing patient's life-style. Little daily insulin requirement < 20 U.l./24 hours, pregnancy. The essential advantages in personal insulin pump use are: prompt metabolic control improvement, a considerable liberty in meal consumption (in its time and amount) and intensity of effort, decrease in number of hypoglycemia and exactly insulin dosage establishment. PMID- 16379331 TI - [Discovering of mucosal immunity]. AB - The mucosal surface area of gastrointestinal, respiratory and urogenital tracts of an adult individuals is about 400 m2 and is the main route of infection and potential harmful agents penetration. It emphasize the essential role of the mucosal immune system (MALT). The beginnings of mucosal immunology, based on the people's observations and believes, descent from ancient times, when the attempt of immunization of human organism against distressed illness have been undertaken. Discovery of a MALT system elements and mechanisms of the mucosal immunity working occured in the 20th century. At that time, the base of beneficial properties of human and other animals' secretions have been recognized, starting from milk, through saliva, ending to feces. Nowadays, it is known, that in all secretions, beside innate defence, the main part of mucosal immunity perform antibodies synthetized locally, with the predominance of the class A (IgA). In this paper the greatest discoveries, leading to the understanding of mucosal immunization resulting in systemic resistance, have been approached. PMID- 16379332 TI - [Role of procalcitonin concentration in clinical diagnosis]. AB - Procalcytonin (PCT) is a prohormon of calcitonin. It is made in thyroid structures. Level of procalcitonin is related from actual calcium concentration in blood. It is shown that procalcitonin level increases in bacterial, fungus and parasite infections, in systematic inflammatory response syndrome (SIRS), burn and multiple organ dysfunction syndrome (MODS). We wanted to conclude the actual knowledge about the role of procalcitonin used in acute infection. Level of PCT is compatible with the etiology of infection. The procalcitonin is a fast infection marker with a longer semi-loose time than CRP. A mechanism of secretion of procalcitonin to blood circulation is still enigmatic. The determination method (immunoluminometric assay) is easy and inexpensive and results are credible and can be used to differentiating and moni-toring of disease. CONCLUSIONS: It is possible that PCT could be an important marker in fast diagnosis and differentiating of bacterial infections, also could be used in treatment monitoring and prognosis. PMID- 16379333 TI - [Effects of atrial fibrillation on clinical outcome of cerebral ischemic stroke]. AB - Atrial fibrillation is one of the most common cardiac arrhythmia met in clinical practice. Nonregular heart rhythm leads to haemodynamic disturbances and favors thromboembolic complications--most frequently ischemic strokes. Cardioembolic strokes have different symptomatology compared with atherothrombotic strokes. They are characterised by more sudden onset, frequent haemorrhagic transformation and worse prognosis. Until now there is no clear explanation of unfavourable course of stroke in these patients. Concomitant diseases and sudden artery closure in subjects with undeveloped collateral circulation are considered as resulting in more extensive infarct size. Also impact of different forms of atrial fibrillation on the course of cerebrovascular events is not established. Paroxysmal and chronic atrial fibrillation have different influence on haemodynamic and hemostatic parameters what may play an important role in course of acute phase of ischemic stroke. Based on the results of clinical and preclinical studies, the set of data concerning the effects of different forms of atrial fibrillation on acute stroke outcome and actual guidelines of cardioembolic acute stroke treatment are presented. PMID- 16379334 TI - [Statins and stroke]. AB - The occurrence of stroke increases with age, particularly affecting the older elderly, a population also at higher risk for coronary heart disease (CHD). Epidemiological and observational studies have not shown a clear association between cholesterol levels and all causes of stroke. Nevertheless, large, long term statin trials in patients with established CHD or et high risk for CHD (diabetes, hypertension) have shown that statins decrease stroke incidence in these populations even with a normal baseline cholesterol concentration. In patients with previous stroke statins reduce the incidence of coronary events, but whether they actually reduce the incidence of recurrent strokes in secondary prevention is unproved. In this review we discuss the potential reason for the effects of statins on stroke and the mechanisms of action. Statins probably reduce stroke by a variety of mechanisms. Several studies indicate that statins have multiple effects beyond lowering the cholesterol level. There is evidence that statins have neuroprotective properties for the acute ischaemic brain. Statins interfere with platelet aggregation and have anti-inflammatory and antioxidative properties. Also statins promote stabilisation of atherosclerotic plaques and improve blood flow to the ischaemic brain. The protective effects of statins might be due to their direct effect on endothelial cells leading to improved nitric oxide (NO) bioavailability. However further studies are needed to understand the full role of statins in the prevention of stroke in patients without established cardiovascular disease, representative of the typical stroke population. PMID- 16379335 TI - [Assessment of the efficacy of cancer procoagulant (CP) activity evaluation in the oncological diagnosis of the urinary system]. AB - Many studies have been carried out to develop unfailing diagnostic methods that could improve cancer detection. There are available cancer markers of relatively low sensitivity and specificity, which makes a reason why they not always let detect neoplasm at their earliest stage. There is a new protease cysteine enzyme named cancer procoagulant (CP) isolated from rabbit V2 Ca neoplasm and characterized by Gordon et al in 1975. Because of its exclusive presentation in the cancer tissues and blood serum of the patients with tumor, this neoplasm-cell originated protein seems to be a new biochemical cancer disease marker. The elevated activity of CP was found in the cancers of pancreatic, breast, lung, alimentary and urinary system. The blood serum CP activity levels in the patients with renal, bladder, and prostate cancers were determined statistically higher as compared to controls but the difference varied depending on the mentioned organ of the urinary system. The CP highest activity levels was determined in the patients with prostate cancer, lower ones in bladder carcinoma ones and the lowest ones in individuals with renal tumours. That is why it appears to be justifiable to apply the determination of the CP in the oncological diagnosis in the urinary system. PMID- 16379336 TI - [Can the administration of large doses of vitamin C have a harmful effect?]. AB - Vitamin C in its pure form or as a component of multivitamin and combined drugs belongs to the over-the-counter drugs that are available not only at the chemist's but in retail shops as well. Intensive promotion of the advantages of vitamin C as the compound that augments immunity to all sorts of infections, accelerates recovery, eliminates the symptoms of common cold and flu-like diseases, and contributes to general well-being, is of great importance to its intake in quantities far exceeding the recommended dietary allowance. Moreover, the subjects who simultaneously take several anti-common cold drugs are not aware of the fact that almost each of them contains ascorbic acid. Among numerous reports concerning positive effect of vitamin C as antioxidant and free radicals scavenger, there are also those pointing at its potentially pernicious effect. Vitamin C, especially in the doses exceeding daily recommended dietary allowance may result in oxalate crystallization, formation of advanced glycation end products and even exert prooxidant effect. PMID- 16379337 TI - Geriatric medical education and training in the United States. AB - Medical education in geriatrics is an important requirement to ready the profession to provide comprehensive health care to the world's and also Taiwan's aging population. The predoctoral curricula and postdoctoral training programs in the United States were developed and supported by government agencies and professional education societies. Geriatric medical education in American medical schools has improved in the past 20 years, yet is still facing many challenges. The purposes of this paper are to review the current progress of, and propose some main principles and policies for the development of geriatric medical education and current progress in the United States. Geriatric medical education should be mandatory to adequately prepare medical students, residents, fellows, and practicing physicians to treat the elderly. The current progress and practice of geriatric medical education at the University of Texas Health Science Center at San Antonio are presented as an example. PMID- 16379338 TI - Plasma hyperhomocysteinemia, MTHFR polymorphism and thromboembolic disease: an example of gene-nutrition interactions in chronic disease. PMID- 16379339 TI - Influence of methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism, B vitamins and other factors on plasma homocysteine and risk of thromboembolic disease in Chinese. AB - BACKGROUND: Thromboembolic disease is a major cause of morbidity and mortality in many countries. Our previous study found that Chinese subjects carried the same polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene as described in Western studies. The aim of the present study was to determine the influence of MTHFR polymorphism, B vitamins and other factors on plasma homocysteine (Hcy) levels and risk of thromboembolic disease in Chinese. METHODS: One hundred and six subjects were enrolled into the study. They were categorized into 4 groups: healthy individuals (n = 42); those with diabetes mellitus (n = 20); those with deep vein thrombosis (DVT) (n = 11); and those with coronary artery disease (CAD) (n = 33). Plasma levels of folic acid, vitamins B6 and B12, Hcy, and fasting blood sugar were measured; total cholesterol, triglycerides, complete blood count, and 677 C-->T mutation in MTHFR were determined. RESULTS: Plasma Hcy was lowest in the healthy subjects, higher in diabetics, followed by patients with DVT, and highest in patients with CAD (p < 0.001, ANOVA). MTHFR C677T polymorphism was the common factor affecting plasma logHcy levels in all 4 groups of subjects. Triglycerides affected plasma logHcy in the CAD patients. For the 4 groups as a whole, MTHFR polymorphism, triglycerides, and vitamin B12 were the most significant factors influencing plasma Hcy. CONCLUSION: We suggest that high plasma Hcy is an important risk factor for CAD. Other factors including MTHFR polymorphism, vitamin B12, triglycerides, total cholesterol, and gender might affect Hcy levels in different diseases and conditions. PMID- 16379340 TI - Relationship between serum leptin levels and body composition and markers of malnutrition in nondiabetic patients on peritoneal dialysis or hemodialysis. AB - BACKGROUND: Leptin is a protein hormone secreted by adipocytes, regulating body fat and food intake. It has been reported that serum leptin levels are high in patients with chronic renal failure, and this fact has been associated with malnutrition and body composition changes in patients on hemodialysis. This present study investigated the relationship between plasma leptin concentrations and body composition and markers of malnutrition in nondiabetic patients diagnosed with end-stage chronic renal failure, treated with continuous ambulatory peritoneal dialysis (CAPD) or hemodialysis (HD). METHODS: A total of 152 HD and 32 CAPD patients were enrolled into the study. The body compositions of the patients were established by utilizing a Body Composition Analyzer. Triceps skinfold thickness (TSFT) was measured by using a Harpenden Skinfold Caliper. Serum leptin level was detected by radioimmunoassay in ng/mL through employing a DPC Gambyt-CR gamma counter. Standard laboratory methods were used for measuring the remaining parameters (total protein, albumin, blood urea nitrogen, creatinine, hemoglobin, hematocrit, high-sensitivity C-reactive protein [hsCRP]). RESULTS: No significant difference was observed between the HD and CAPD groups regarding leptin levels. Leptin levels of female patients in both groups were markedly higher when compared with those of men (p = 0.001). Plasma leptin levels in total, as well as for both male and female HD and CAPD patients, significantly correlated positively with age, percent fat, fat mass, body mass index and TSFT (p = 0.001). Serum leptin levels were not found to be correlated with length of time on dialysis, lean body mass, total body water, hsCRP, total protein and albumin levels (p > 0.05). CONCLUSION: The data obtained in this study indicated that serum leptin levels could be instrumental markers in establishing body fat ratio, as well as in determining metabolic and nutritional factors in patients with chronic renal failure. PMID- 16379341 TI - Epidemiologic and clinical features of measles and rubella in a rural area in China. AB - BACKGROUND: Changes in the epidemiologic and clinical features of measles have resulted in diagnostic difficulties in distinguishing between measles and other rash and fever illnesses such as rubella. The objectives of this article are to report the true morbidity and epidemiologic and clinical features of measles and rubella, provide objective diagnostic indicators for distinguishing between the 2 diseases, and provide a time-efficient scientific basis for accurately evaluating the effectiveness of immunization with the measles vaccine and regulating the control strategies for measles in the rural areas of China in the cold chain era. METHODS: From 1992 to 2002, patients with measles and rubella were recorded and reported routinely in the rural disease surveillance spots of Jingzhou District, Hubei Province, by collecting their sera and testing for measles and rubella IgM and IgG antibodies. RESULTS: Over the 11-year study period, 146 cases of measles were reported in the disease surveillance spots. The annual incidence of measles was highest in 1999 (118.8/100,000 population). The incidence of measles in the 4 6 and 7-15 year old age groups were higher than in other age groups. During 5 outbreaks of measles, a total of 128 cases were reported, accounting for 87.7% of the total number of measles cases. A total of 566 rubella cases were reported, with 3 epidemic peaks (in 1992, 1997 and 2002). The incidence of rubella in the 4 6 and 7-15 year old age groups were higher than in other age groups. There were 13 outbreaks of rubella, with 547 cases being reported, accounting for 96.6% of the total number of rubella cases. Accuracy of the clinical diagnosis of measles was 40%, while it was 100% for rubella. The proportions of patients who had a moderate to high fever (> or = 38.6 degrees C) in the measles group (89.5%) and the combined infection (both measles and rubella) group (90%) were significantly higher than that of the rubella group (30.6%). The proportions of patients whose prodromal stage lasted > or = 3 days in the former 2 groups (68.4% and 95%, respectively) were significantly higher than that of the rubella group (16.7%). The proportions of patients whose exanthema lasted > or = 4 days in the former 2 groups (78.9% and 75%, respectively) were significantly higher than that of the rubella group (26.9%). CONCLUSION: Measles and rubella are 2 common communicable diseases in the children of Jingzhou District, with outbreaks being the main epidemic form. Some clinical features such as temperature, duration of the prodromal stage and exanthema are different in measles and rubella, and they can be useful in distinguishing between the 2 diseases. Accuracy in the clinical diagnosis of measles should be improved by enhancing serologic testing for measles and rubella, and by identifying patients' specific clinical characteristics. PMID- 16379342 TI - Endovascular treatment of ruptured vertebral dissecting aneurysms with electrodetachable coils. AB - BACKGROUND: Ruptured intradural vertebral dissecting aneurysm (VDA) is associated with a high risk of rebleeding and a high incidence of mortality if left untreated because of its natural history. We report our experience of endovascular treatment of ruptured VDAs using electrodetachable coils. METHODS: Over 7 years, 10 patients with spontaneously ruptured intradural VDAs were managed by endovascular embolization in our institute. All patients received endovascular trapping of the dissecting aneurysm and proximal occlusion by electrodetachable coils. RESULTS: There were 5 men and 5 women, with a mean age of 48 years. Of the 10 VDAs, 8 were located in the supra-posterior inferior cerebellar artery (PICA) area, and 2 were in the infra-PICA area. Six affected vertebral arteries were dominant (n = 3) or co-dominant (n = 3) in the vertebrobasilar system. Nine ruptured VDAs and their parent arteries were successfully occluded by a single session of endovascular embolization by electrodetachable coils. In the tenth patient, the dissecting aneurysm showed subtotal occlusion because of involvement of the vertebrobasilar junction. The mean number and coil length were 7 and 58 cm, respectively. There were no significant periprocedural complications or recurrent bleeding. All patients demonstrated good clinical recovery (n = 9) or improvement (n = 1) after embolization over a mean follow-up of 15 months (range, 6-24 months). CONCLUSION: Endovascular embolization is a useful and safe method in the treatment of ruptured VDAs even when the affected vertebral arteries are dominant or co dominant in vertebrobasilar flow. Trapping procedures and direct occlusion of the dissecting segment with preservation of the PICA by detachable coils should be performed as early as possible in the management of VDAs. PMID- 16379343 TI - Klinefelter's syndrome with seizure, pseudohypoparathyroidism type Ib and multiple endocrine dysfunctions. AB - Klinefelter's syndrome is rarely associated with hypocalcemia, especially pseudohypoparathyroidism (PHP) type Ib. We describe a case of Klinefelter's syndrome associated with seizure, PHP type Ib and multiple endocrine dysfunctions. A 19-year-old Taiwanese male was admitted due to seizures with loss of consciousness. He had been diagnosed with Klinefelter's syndrome with seizure disorder and hypocalcemia 3 months previously. Physical examination revealed eunuchoidism but no osteodystrophy, while laboratory data revealed severe hypocalcemia, hyperphosphatemia, and elevated parathyroid hormone. Chromosomal study showed 47,XXY. Osteoporosis was found on chest and abdominal radiography. Dense calcification in the cerebrum and cerebellum was shown on brain computed tomography and magnetic resonance imaging. Elevation of the patient's serum calcium level was noted after vitamin D and calcium carbonate supplements were given. Klinefelter's syndrome is rarely associated with PHP type Ib; our patient's hypocalcemia improved after long-term aggressive treatment. PMID- 16379344 TI - Acupuncture resulting in immediate bronchodilating response in asthma patients. AB - There are some encouraging results in the English literature that show acupuncture resulting in an immediate improvement in pulmonary function, but there are also studies that have not demonstrated any benefit. We present 3 patients with persistent asthma who experienced immediate bronchodilatation after acupuncture without the use of any short-acting bronchodilator. After needle stimulation on selected acupoints, clinical symptoms such as dyspnea and wheezing improved. Pulmonary function test showed immediate improvement in forced expiratory volume in 1 second (FEV1), more than 20% as compared with baseline FEV1. Pulmonary function returned to baseline within 4 hours after acupuncture in 2 patients. From our observations of these 3 asthma patients, acupuncture may improve clinical dyspnea symptoms and performance on pulmonary function tests. Further large-scale controlled studies should be conducted to determine the effectiveness of acupuncture in the treatment of asthma. PMID- 16379345 TI - Laparoscopic detorsion of twisted ovary. AB - An 18-year-old female presented with a 6-day history of lower abdominal pain. Transabdominal ultrasonography revealed a right adnexal cyst measuring 9 cm. Laparoscopic examination demonstrated a 10-cm right paratubal cyst and a 5-cm right ovarian torsion with gangrenous discoloration. After removal of the right paratubal cyst, untwisting of the right ovary was performed. The postoperative period was uneventful. During follow-up, ultrasonography revealed restoration of right ovarian size with follicular growth. Arterial and venous blood flows were seen on Doppler examination. The right ovary appeared to be completely viable through a second-look laparoscopic examination. A twisted ovary may be completely restored with conservative management. PMID- 16379346 TI - Diagnosis of appendicitis with left lower quadrant pain. AB - Abdominal pain is one of the most common chief complaints of patients presenting to the emergency department and, among the diagnoses of abdominal pain, appendicitis is the most common surgical disorder. Traditionally, the diagnosis of appendicitis is based on well-established clinical criteria combined with physician experience. However, appendicitis presenting with rare and misleading left lower quadrant (LLQ) pain may result in an initial false-negative diagnosis by the physician and even result in failure to order the subsidiary examination of computed tomography (CT) or ultrasound, so increasing the risk of perforation/abscess formation and prolonged hospital stay. In this report, we present 2 cases of atypical appendicitis with LLQ pain where the correct diagnosis was not initially considered. One patient had right-sided appendicitis; the inflamed appendix was 12 cm in length and projected into the LLQ. Local peritonitis developed during observation. With the aid of CT, the diagnosis was established in time. The other patient had left-sided appendicitis with situs inversus totalis. Adverse outcomes with appendiceal rupture and abscess formation occurred due to inadvertent physical examinations and inadequate observation. Early clinical suspicion and adequate observation are indicated in patients with uncertain clinical features. However, in patients with unresolved clinical symptoms and/or local peritonitis that develop during observation, imaging studies play a significant role in preoperative diagnosis and determination of proper treatment. PMID- 16379347 TI - We need to cool it. PMID- 16379348 TI - And the beat goes on. PMID- 16379349 TI - Now available: middle ground. PMID- 16379350 TI - Senate budget language calls for permanent self-referral moratorium, "75 percent rule" threshold extension, P4P. PMID- 16379351 TI - Federation/AHA report cites significant "75 percent rule" limits to rehab care. PMID- 16379352 TI - Beacons of help and hope. PMID- 16379353 TI - Our employees showed great courage. PMID- 16379354 TI - Tenet Healthcare Corporation--Hurricane Katrina response. PMID- 16379355 TI - A new standard of dedication and commitment. PMID- 16379356 TI - Kindred Healthcare Inc. response to Hurricane Katrina. PMID- 16379357 TI - Truly a team effort. PMID- 16379358 TI - Wesley Medical Center's response to Hurricane Katrina. PMID- 16379359 TI - Summit Hospital's response to Hurricane Katrina. PMID- 16379360 TI - Surviving Katrina. PMID- 16379361 TI - Baptist Health System (San Antonio, TX)--response to Hurricanes Katrina and Rita. PMID- 16379362 TI - Improving the nation's health care system--beginning in New Orleans. PMID- 16379363 TI - Overcoming compassion fatigue. PMID- 16379365 TI - Sebelius denies BCBSK aquisition by Anthem. PMID- 16379366 TI - Aligning biological sequences on distributed bus networks: a divisible load scheduling approach. AB - In this paper, we design a multiprocessor strategy that exploits the computational characteristics of the algorithms used for biological sequence comparison proposed in the literature. We employ divisible load theory (DLT) that is suitable for handling large scale processing on network based systems. For the first time in the domain of DLT, the problem of aligning biological sequences is attempted. The objective is to minimize the total processing time of the alignment process. In designing our strategy, DLT facilitates a clever partitioning of the entire computation process involved in such a way that the overall time consumed for aligning the sequences is a minimum. The partitioning takes into account the computation speeds of the nodes and the underlying communication network. Since this is a real-life application, the post-processing phase becomes important, and hence we consider propagating the results back in order to generate an exact alignment. We consider several cases in our analysis such as deriving closed-form solutions for the processing time for heterogeneous, homogeneous, and networks with slow links. Further, we attempt to employ a multiinstallment strategy to distribute the tasks such that a higher degree of parallelism can be achieved. For slow networks, our strategy recommends near optimal solutions. We derive an important condition to identify such cases and propose two heuristic strategies. Also, our strategy can be extended for multisequence alignment by utilizing a clustering strategy such as the Berger Munson algorithm proposed in the literature. Finally, we use real-life DNA samples of house mouse mitochondrion (Mus Musculus Mitochondrion, NC_001569) consisting of 16,295 residues and the DNA of human mitochondrion (Homo Sapiens Mitochondrion, NC_001807) consisting of 16,571 residues, obtainable from the GenBank, in our rigorous simulation experiments to illustrate all the theoretical findings. PMID- 16379367 TI - Prediction of care class by local additive reference to prototypical examples. AB - The public long-term care insurance program for the elderly in Japan set out in 2000 toward establishing a new system whereby citizens can be assured that they will receive care and be supported by the society as a whole. The insurance program includes computer-aided certification processes to estimate the needs for nursing care for clients. In this work, we show the applicability of an adaptive local nonlinear approximation method associated with the Japanese national database for automatic inference of the care class. PMID- 16379368 TI - Automated DNA fragments recognition and sizing through AFM image processing. AB - This paper presents an automated algorithm to determine DNA fragment size from atomic force microscope images and to extract the molecular profiles. The sizing of DNA fragments is a widely used procedure for investigating the physical properties of individual or protein-bound DNA molecules. Several atomic force microscope (AFM) real and computer-generated images were tested for different pixel and fragment sizes and for different background noises. The automated approach minimizes processing time with respect to manual and semi-automated DNA sizing. Moreover, the DNA molecule profile recognition can be used to perform further structural analysis. For computer-generated images, the root mean square error incurred by the automated algorithm in the length estimation is 0.6% for a 7.8 nm image pixel size and 0.34% for a 3.9 nm image pixel size. For AFM real images we obtain a distribution of lengths with a standard deviation of 2.3% of mean and a measured average length very close to the real one, with an error around 0.33%. PMID- 16379369 TI - Evaluation of statistical association measures for the automatic signal generation in pharmacovigilance. AB - Pharmacovigilance aims at detecting the adverse effects of marketed drugs. It is generally based on the spontaneous reporting of events thought to be the adverse effects of drugs. Spontaneous Reporting Systems (SRSs) supply huge databases that pharmacovigilance experts cannot exhaustively exploit without data mining tools. Data mining methods; i.e., statistical association measures in conjunction with signal generation criteria, have been proposed in the literature but there is no consensus regarding their applicability and efficiency, especially since such methods are difficult to evaluate on the basis of actual data. The objective of this paper is to evaluate association measures on simulated datasets obtained with SRS modeling. We compared association measures using the percentage of false positive signals among a given number of the most highly ranked drug-event combinations according to the values of the association measures. By considering 150 drugs and 100 adverse events, these percentages of false positives, among the 500 most highly ranked drug-event couples, vary from 1.1% to 53.4% (averages over 1000 simulated datasets). As the measures led to very different results, we could identify which measures appeared to be the most relevant for pharmacovigilance. PMID- 16379371 TI - Knowledge representation and sharing using visual semantic modeling for diagnostic medical image databases. AB - Information technology offers great opportunities for supporting radiologists' expertise in decision support and training. However, this task is challenging due to difficulties in articulating and modeling visual patterns of abnormalities in a computational way. To address these issues, well established approaches to content management and image retrieval have been studied and applied to assist physicians in diagnoses. Unfortunately, most of the studies lack the flexibility of sharing both explicit and tacit knowledge involved in the decision making process, while adapting to each individual's opinion. In this paper, we propose a knowledge repository and exchange framework for diagnostic image databases called "evolutionary system for semantic exchange of information in collaborative environments" (Essence). This framework uses semantic methods to describe visual abnormalities, and offers a solution for tacit knowledge elicitation and exchange in the medical domain. Also, our approach provides a computational and visual mechanism for associating synonymous semantics of visual abnormalities. We conducted several experiments to demonstrate the system's capability of matching synonym terms, and the benefit of using tacit knowledge in improving the meaningfulness of semantic queries. PMID- 16379370 TI - A multiagent system enhancing home-care health services for chronic disease management. AB - In this paper, a multiagent system (MAS) is presented, aiming to enhance monitoring, surveillance, and educational services of a generic medical contact center (MCC) for chronic disease management. In such a home-care scenario, a persistent need arises for efficiently monitoring the patient contacts and the MCC's functionality, in order to effectively manage and interpret the large volume of medical data collected during the patient sessions with the system, and to assess the use of MCC resources. Software agents were adopted to provide the means to accomplish such real-time information-processing tasks, due to their autonomous, reactive and/or proactive nature, and their effectiveness in dynamic environments by incorporating coordination strategies. Specifically, the objective of the MAS is to monitor the MCC environment, detect important cases, and inform the healthcare and administrative personnel via alert messages, notifications, recommendations, and reports, prompting them for actions. The main aim of this paper is to present the overall design and implementation of a proposed MAS, emphasizing its functional model and architecture, as well as on the agent interactions and the knowledge-sharing mechanism incorporated, in the context of a generic MCC. PMID- 16379372 TI - Tailored reversible watermarking schemes for authentication of electronic clinical atlas. AB - It is accepted that digital watermarking is quite relevant in medical imaging. However, due to the special nature of clinical practice, it is often required that watermarking not introduce irreversible distortions to medical images. The electronic clinical atlas has such a need of "lossless" watermarking. We present two tailored reversible watermarking schemes for the clinical atlas by exploiting its inherent characteristics. We have implemented the schemes and our experimental results look very promising. PMID- 16379373 TI - An intelligent remote monitoring system for artificial heart. AB - A web-based database system for intelligent remote monitoring of an artificial heart has been developed. It is important for patients with an artificial heart implant to be discharged from the hospital after an appropriate stabilization period for better recovery and quality of life. Reliable continuous remote monitoring systems for these patients with life support devices are gaining practical meaning. The authors have developed a remote monitoring system for this purpose that consists of a portable/desktop monitoring terminal, a database for continuous recording of patient and device status, a web-based data access system with which clinicians can access real-time patient and device status data and past history data, and an intelligent diagnosis algorithm module that noninvasively estimates blood pump output and makes automatic classification of the device status. The system has been tested with data generation emulators installed on remote sites for simulation study, and in two cases of animal experiments conducted at remote facilities. The system showed acceptable functionality and reliability. The intelligence algorithm also showed acceptable practicality in an application to animal experiment data. PMID- 16379374 TI - Meeting the challenges of oral health for older people: a strategic review. PMID- 16379375 TI - Electrochemistry and electroanalytical applications of carbon nanotubes: a review. AB - This review addresses recent developments in electrochemistry and electroanalytical chemistry of carbon nanotubes (CNTs). CNTs have been proved to possess unique electronic, chemical and structural features that make them very attractive for electrochemical studies and electrochemical applications. For example, the structural and electronic properties of the CNTs endow them with distinct electrocatalytic activities and capabilities for facilitating direct electrochemistry of proteins and enzymes from other kinds of carbon materials. These striking electrochemical properties of the CNTs pave the way to CNT-based bioelectrochemistry and to bioelectronic nanodevices, such as electrochemical sensors and biosensors. The electrochemistry and bioelectrochemistry of the CNTs are summarized and discussed, along with some common methods for CNT electrode preparation and some recent advances in the rational functionalization of the CNTs for electroanalytical applications. PMID- 16379376 TI - Sensitive and selective analysis of polychlorinated dibenzo-p dioxins/dibenzofurans and their precursors by supersonic jet/resonance-enhanced multiphoton ionization/time-of-flight mass spectrometry. AB - Polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs), some of which are extremely toxic, are present at low concentrations in incinerator flue gases. The on-line real-time measurement of these compounds is necessary, since these concentrations fluctuate drastically with the process conditions used. Supersonic jet/resonance-enhanced multiphoton ionization/time-of-flight mass spectrometry (SSJ/REMPI/TOF-MS), which provides both high selectivity and sensitivity, appears suitable for application to the on-line real-time monitoring of PCDD/Fs. In this review, the resonant ionization wavelengths of PCDD/Fs and their precursors are given. Moreover, improvements in SSJ/REMPI/TOF-MS that lead to enhanced sensitivity and selectivity are presented. That is, novel ionization sources and advanced ionization schemes are reported for the sensitive measurement of PCDD/Fs and their precursors. Those techniques are advantageous for the trace analysis of these compounds. PMID- 16379377 TI - Mechanism of volatile hydride formation and their atomization in hydride generation atomic absorption spectrometry. AB - The mechanism of volatile hydride generation (HG) and the formation of analyte atoms in the quartz cell atomizer used in the determination of hydride-forming elements (As, Bi, Ge, Pb, Sb, Sn, Te etc.) by atomic absorption spectrometry (AAS), have been critically reviewed. The nascent hydrogen mechanism failed to explain hydride generation under different experimental conditions when tetrahydroborate (THB), amineboreanes (AB) and cyanotrihydroborate (CBH) were used as reductants. Various experimental evidence suggested a non-nascent hydrogen mechanism, in which the transfer of hydrogen directly bonded to boron to an analyte takes place. In electrochemical hydride generation (EcHG), the reduction of the analyte species and subsequent hydrogenation was proposed. The mechanism of analyte atom formation in a quartz tube atomizer has been explained by the following hypotheses: thermal decomposition, oxidation by 02 and collisions by hydrogen free radicals. The free-radical mechanism satisfactorily explains most of the analytical implications. The significant variation in the experimental conditions required to generate different analyte hydrides makes it difficult to arrive at a generalized mechanism of hydride formation. PMID- 16379378 TI - Laser ablation and low-pressure helium-ICP-MS for the analysis of alumina powder dispersed in glycerol. AB - A combined method of laser ablation (LA) and ICP-MS has gained much attention as a direct analytical method for solid samples. The determination of some elements, however, is seriously disturbed by isobaric interferences, mainly caused by argon and ambient air constituents. The use of low-pressure helium-ICP is a promising solution of the problem. A 1:1 mixture of alumina powder and glycerol was deaerated and irradiated with a pulsed laser beam (150 mJ) for 10 s. The sample aerosol was transported to the ICP with a stream of helium. Indium was used as an internal standard for correcting the ablated sample amount. Calibration curves were prepared from glycerol containing high-purity alumina, trace metals and indium. The detection limits for Cr, Mn, Fe, Co, Ni, and Cu approached the fractional ppm levels. The proposed method was successfully applied to the analysis of different alumina samples (99 - 99.995% purity). PMID- 16379379 TI - Development of rotating liquid membrane disk electrode and rotating liquid membrane ring-liquid membrane disk electrode and evaluation of characteristics of ion transfer reactions at the rotating aqueous/organic solution interface. AB - A liquid membrane disk electrode, LMDE, and a liquid membrane ring-liquid membrane disk electrode, LMRE-LMDE, were developed by placing a gelled polyvinyl chloride thin membrane impregnated with 2-nitrophenyl octyl ether, NPOE-LM, on the surface of a glassy carbon, GC, disk or ring electrode. The voltammogram for the ion transfer at the interface between an aqueous solution, W, and NPOE-LM was recorded by setting the developed electrode in W and rotating at a rate, omega, between 0 and 4000 rpm. The sensitivity of the ion-transfer current at the WINPOE LM interface, I, was enhanced to be more than 100 times better than that at the WINPOE (solution) interface when LMDE was rotated at omega higher than 200 rpm. The reversibility of the ion transfer reaction could be evaluated based on the dependence of I on omega of LMDE, and the reaction product at LMDE could be identified at LMRE when the rotating LMRE-LMDE system was adopted. PMID- 16379380 TI - The detection of nitrate using in-situ copper nanoparticle deposition at a boron doped diamond electrode. AB - Electrochemical deposition from a 0.1 M sodium sulphate solution, containing Cu2+ (adjusted to pH 3 with hydrochloric acid) produced a well defined copper nanoparticle deposit on the surface of a boron doped diamond electrode. Changing conditions such as potential (-0.8, -1.0 and -1.2 V), time (5, 2 and 0.5 s) and concentration of Cu2+ (500, 250 and 100 microM) was found to give copper nanoparticles of varying size and particle density. The electrocatalytic properties of the copper surface towards nitrate reduction were explored. An in situ copper nanoparticle production method was developed for the detection of nitrate; this involves electrodeposition, followed by linear sweep voltammetry for the reduction of nitrate and then application of a stripping potential to renew the electrode surface. The linear sweep was discovered to have homogenised the size of the nanoparticles but their number density was still dependant on the initial conditions of deposition. Some particles were still present at the surface after the stripping potential had been applied but repetitions of the procedure showed these did not have an effect on subsequent deposits. Optimisation of the method lead to applying a deposition potential of -0.8 V, at a BDD electrode for 5 s in a 0.1 M sodium sulphate solution (pH 3) containing 100 microM Cu2+ followed by a linear sweep at 1 V/s; this yielded a limit of detection of 1.5 microM nitrate. The analytical applicability of the technique was evaluated for nitrate detection in a natural mineral water sample and was found to agree well with that stated by the manufacturer. PMID- 16379381 TI - A surface plasmon resonance sensor for substance P using gold-modified calmodulin and melittin. AB - A surface plasmon resonance (SPR) biosensor for the quantification of a neuropeptide substance P (SP) is described based on an inhibition assay using Au colloid-modified calmodulin (Au-CaM) and a target peptide melittin immobilized on carboxymethylated dextran. The modification of CaM with streptavidin Au colloids was achieved in a sample solution by the amine coupling method. The SPR signal sharply increased, corresponding to the formation of a Ca2+-Au-CaM-melittin complex on the sensor surface, and approached a steady state within 5 min. When SP was added to a sample solution, the SPR signal was decreased, due to the formation of a Ca2+-Au-CaM-SP complex in the sample solution. The modification of CaM with streptavidin Au colloids was effective for enhancing the SPR signal for SP. A decrease in the SPR signal was observed for SP in the concentration range from 0.10 to 5.0 microM, whose lower limit was ten-times superior to that (1.0 microM) with unmodified CaM. The response was highly selective to SP and the selectivity was in the order of SP >> neurokinin A > neurokinin B > neurotransmitters (glycine, GABA, L-glutamate, acetylcholine, norepinephrine, 5HT) - substance P fragment (1 - 7). The potential use of the present sensor for the quantification of SP in mouse brain extracts is demonstrated. PMID- 16379382 TI - Genotyping of the human lipoprotein lipase gene by ferrocenylnaphthalene diimide based electrochemical hybridization assay. AB - A ferrocenylnaphthalene diimide (FND)-based electrochemical hybridization assay (FND-EHA) was applied to the detection of two mutations in human lipoprotein lipase (LPL) gene, G188E (one base transition) and Arita (one base deletion). A probe oligodeoxyribonucleotide of 13 bases representing the wild type (WT) sequence of LPL was immobilized on a gold electrode, followed by hybridization with a sample PCR product of 350 base pairs under conditions in which both WT and mutated (MT) sequences could form a duplex with the probe. The hybridized electrodes were soaked in an electrolyte containing FND under conditions in which only the mismatched duplex could undergo dissociation. FND was concentrated in proportion to the amount of the duplex remaining on the electrode to give rise to a current signal. Blind tests were run to judge the genotype (WT/WT, WT/MT, or MT/MT) of 10 samples each for the G188E and Arita mutations and then, 8 and 10 of them were judged correctly, respectively. PMID- 16379383 TI - Ion-selective electrodes for potentiometric determination of ranitidine hydrochloride, applying batch and flow injection analysis techniques. AB - New ranitidine hydrochloride (RaCl)-selective electrodes of the conventional polymer membrane type are described. They are based on incorporation of ranitidine-tetraphenylborate (Ra-TPB) ion-pair or ranitidine-phosphotungstate (RaPT) ion-associate in a poly(vinyl chloride) (PVC) membrane plasticized with dioctylphthalate (DOP) or dibutylphthalate (DBP). The electrodes are fully characterized in terms of the membrane composition, solution temperature, and pH. The sensors showed fast and stable responses. Nernstian response was found over the concentration range of 2.0 x 10(-5) M to 1.0 x 10(-2) M and 1.0 x 10(-5) M to 1.0 x 10(-2) M in the case of Ra-TPB electrode and over the range of 1.03 x 10( 5) M to 1.00 x 10(-2) M and 1.0 x 10(-5) M to 1.0 x 10(-2) M in the case of Ra-PT electrode for batch and FIA systems, respectively. The electrodes exhibit good selectivity for RaCl with respect to a large number of common ions, sugars, amino acids, and components other than ranitidine hydrochloride of the investigated mixed drugs. The electrodes have been applied to the potentiometric determination of RaCl in pure solutions and in pharmaceutical preparations under batch and flow injection conditions with a lower detection limit of 1.26 x 10(-5) M and 5.62 x 10(-6) M at 25 +/- 1 degrees C. An average recovery of 100.91% and 100.42% with a relative standard deviation of 0.72% and 0.53% has been achieved. PMID- 16379384 TI - Oxidation of ferrocene derivatives in desorption/lonization on porous silicon. AB - In matrix-assisted laser desorption/ionization (MALDI), the true molecular structures of some analytes are not represented by the observed ions due to a redox reaction. In earlier reports, electron transfer from analyte to chemical matrix has been proposed for the oxidation of ferrocene derivatives in MALDI. To address such a redox phenomenon in laser desorption/ionization processes, two ferrocene derivatives, FcCH2CH2Fc and FcCH2NMe2 [Fc:(CsHs)Fe(CsH4)], were analyzed by a matrix-free method, desorption/ionization on porous silicon (DIOS). The oxidized species, Fc+CH2NMe2 and FcCH2CH2Fc+, were detected in the DIOS mass spectra. The results suggested that electron transfer from the analytes to the sample target occurs during the ionization process. PMID- 16379385 TI - Determination of isotopic ratio of boron in boric acid using laser mass spectrometry. AB - A reflectron time-of-flight mass spectrometer (RTOFMS) with a laser ablation ion source was used to determine the isotopic ratio of 10B/11B present in boric acid solutions, using graphite as the matrix. The atom % 10B values obtained were within +/-1% of the actual ones. The determination of the boron isotopic ratio using this method is, in comparison to other methods, faster, less expensive and easier to perform. The results are compared with the values determined using thermal ionization mass spectrometry. PMID- 16379386 TI - Rapid discrimination of fatty acid composition in fats and oils by electrospray ionization mass spectrometry. AB - Fatty acids in 42 types of saponified vegetable and animal oils were analyzed by electrospray ionization mass spectrometry (ESI-MS) for the development of their rapid discrimination. The compositions were compared with those analyzed by gas chromatography-mass spectrometry (GC-MS), a more conventional method used in the discrimination of fats and oils. Fatty acids extracted with 2-propanol were detected as deprotonated molecular ions ([M-H]-) in the ESI-MS spectra of the negative-ion mode. The composition obtained by ESI-MS corresponded to the data of the total ion chromatograms by GC-MS. The ESI-MS analysis discriminated the fats and oils within only one minute after starting the measurement. The detection limit for the analysis was approximately 10(-10) g as a sample amount analyzed for one minute. This result showed that the ESI-MS analysis discriminated the fats and oils much more rapidly and sensitively than the GC-MS analysis, which requires several tens of minutes and approximately 10(-9) g. Accordingly, the ESI MS analysis was found to be suitable for a screening procedure for the discrimination of fats and oils. PMID- 16379387 TI - Retrospective analyses of atmospheric polycyclic and nitropolycyclic aromatic hydrocarbons in an industrial area of a western site of Japan. AB - Thirteen polycyclic aromatic hydrocarbons (PAHs) and four nitropolycyclic aromatic hydrocarbons (NPAHs) on the surfaces of airborne particulates, which were collected at an industrial area of a western site of Japan during periods from 1976 to 1998, were retrospectively analyzed. PAHs and NPAHs were extracted from airborne particulates using hexane with ultrasonication, and then analyzed by HPLC systems with fluorescence detection and chemiluminescence detection, respectively. The total concentrations (mean +/- SD, n = 34) were 15.54 +/- 21.24 ng/m3 for PAHs and 5.85 +/- 8.16 pg/m3 for NPAHs. The concentrations of PAHs and NPAHs were found to be highest during the period between 1979 and 1982, and then reduced. The annual concentrations of PAHs and NPAHs were highly correlated with those of air pollutants from motor vehicle origin, such as carbon monoxide, suspended particulates and non-methane hydrocarbons. The results suggested that motor vehicle emissions were one of the predominant sources of atmospheric PAHs and NPAHs. PMID- 16379388 TI - Identification of ammonia in gas emanated from human skin and its correlation with that in blood. AB - Identifying and measuring the ammonia gas that emanates from human skin, which we called skin gas, has been achieved using a modified gas chromatographic system with a nitrogen-selective detector (flame-thermoionic detector: FTD). The skin gas is collected with a home-made sampling probe or bag, which is used to cover the skin surface of a subject's wrist, or a finger, for 5 min. It was proved that ammonia was present in skin gas for healthy persons and patients with hepatic disease. The average amounts of ammonia were 1.7 +/- 0.4 and 2.7 +/- 0.8 ng/cm2; furthermore, there was a significant difference between them (p < 0.05). In addition, the ammonia levels present in skin gas were correlated with that in blood (r = 0.64, p < 0.05). PMID- 16379389 TI - Prediction of the lifetime of nitrile-butadiene rubber by FT-IR. AB - A quantitative measurement method with FT-IR was proposed for a thermal degradation analysis of nitrile-butadiene rubber (NBR). An NBR film was prepared as a model sample on a barium fluoride (BaF2) crystal plate, which was subjected to a heat treatment. The absorbances of various functional groups were measured directly by FT-IR after thermal degradation at high temperatures. By measuring the absorbances, it was possible to readily determine quantitatively each of the functional groups after the degradation of NBR. By assuming that the NBR lifetime was the point at which the absorbance of a carbon-carbon double bond reaches 45% of that prior to thermal treatment, a method for predicting the lifetime of NBR heated below 150 degrees C was proposed, by using an Arrhenius plot of the heating time versus heating temperature. PMID- 16379390 TI - Determination of the termination efficiency of the transcription terminator using different fluorescent profiles in green fluorescent protein mutants. AB - An approach in determining the intrinsic termination efficiency (%T) of transcription termination using green fluorescent protein (GFP) mutants was developed. This approach utilizes a cassette vector in which the tested terminator is introduced between two GFP mutant genes: an ultraviolet-optimized mutant (GFPuv: F99S, M153T, V163A) and a blue-shifted mutant (BFP: F64L, S65T, T145F). The ratio of the fluorescence intensity of BFP to GFPuv after transcription and translation represents the termination efficiency of the terminator. E. coli ribosomal RNA operon T1 terminator, phage lambda terminator site R2, E. coli tryptophane attenuater were introduced into the vector, and their transcriptional efficiencies were estimated as 89, 79, and 24%, respectively, showing good agreement with published data. PMID- 16379391 TI - GC identification of organic compounds based on partition coefficients of their TMS derivatives in a hexane-acetonitrile system and retention indices. AB - This paper reports the experimentally determined and calculated partition coefficients (Kp) in a heterogeneous system consisting of two partially mixing organic solvents for trimethylsilyl derivatives of 119 compounds including phenols, phenolcarboxylic acids, mono- and dicarboxylic acids, and some carbohydrates and polyols. It was demonstrated that a simple model based on the available physico-chemical properties of molecules can be used to accurately predict the hexane-acetonitrile partition coefficients. Phenols, other organic acids, and carbohydrates form complex mixtures in extracts of plant tissues, in various biological fluids, as well as in waste waters. For group identification of these mixture components, it is desirable to use the parameterj which is a combination of Kp and gas chromatographic retention indices: j = kI - log Kp. This approach increases identification reliability in gas chromatographic analysis. The use of the group parameterj in the analysis of nonvolatile organic components of waste waters is reported. PMID- 16379392 TI - Selective extraction of calcium on tri-n-butyl phosphate plasticized selective extraction of calcium on tri-n-butyl phosphate plasticized polyurethane foam for its spectrophotometric determination in glass and ceramics. AB - The present paper describes the application of a solid phase extraction system in order to separate traces of calcium from glass and ceramics for its spectrophotometric determination. The method is based on the extraction of calcium from sodium hydroxide solution by tri-n-butyl phosphate (TBP) loaded polyurethane foam (PUF), followed by its elution in hydrochloric acid. The spectrophotometric measurement of the absorbance of calcium complex with calconcarboxylic acid (2-hydroxy-1-(2-hydroxy-4-sulfo-1-naphthylazo)-3-naphthoic acid) takes place at pH 12. The following parameters were studied: effects of sodium hydroxide concentration and temperature on the extraction of calcium, time of equilibration for quantitative calcium extraction, effect of TBP concentration, effect of hydrochloric acid concentration for quantitative elution of calcium from PUF, effect of pH and concentration of calconcarboxylic acid for quantitative formation of the complex with calcium, effect of acetone on the stability of calcium-calconcarboxylic acid complex and influence of diverse ions on calcium sorption by TBP-loaded PUF. The results show that calcium traces can be separated onto TBP-loaded PUF from 0.25 mol L(-1) NaOH at 30 +/- 5 degrees C within 30 min. PUF was loaded with TBP in CCl4 (40% v/v). Elution of calcium was done in 1.0 mol l(-1) HCl. The calcium formed a complex with calconcarboxylic acid at pH 12 and absorbance was measured at 560 nm in acetone-water medium. Molar absorptivity was found to be 1.082 x 10(4) l mol(-1) cm(-1). The method obeys Beer's law from 0.10 to 5.0 microg ml(-1) Ca. The validity of the method was established by its successful application in NIST standard reference materials. The method proposed was applied to determine calcium in glass and ceramic materials. The results of the proposed method are comparable with the results of ICP-AES analysis and they are found to be in good agreement. PMID- 16379393 TI - Development of HPLC methods for the determination of cyadox and its main metabolites in goat tissues. AB - Rapid and sensitive high-performance liquid-chromatographic methods were developed for the determination of cyadox, an antimicrobial growth-promoter, and its main metabolites (1,4-bisdesoxycyadox, quinoxaline-2-carboxylic acid) in goat muscle, liver, kidney and fat. Cyadox (CYX) and 1,4-bisdesoxycyadox (BDCYX) in fat were extracted with acetonitrile, and in other tissues with ethyl acetate. Quinoxaline-2-carboxylic acid (QCA) was isolated from tissue hydrolysates by solvent extraction, cleaned up with ion-exchange chromatography, followed by a final liquid-liquid extraction step. UV detections of CYX, BDCYX and QCA were performed at 305, 280 and 320 nm, respectively. The average recoveries of CYX, BDCYX and QCA in spiked tissues at levels of 25, 50, 100 microg/kg were 65 - 92%. The inter-day relative standard deviation for three compounds in different tissues was 5 - 16%. The quantitation limit was 25 microg/kg, and the detection limit was 15 microg/kg for three compounds in various tissues. Incurred goat tissues were analyzed to demonstrate the validity of the described methodologies. The present methods were highly selective and could be used in the metabolism and residue studies of cyadox. PMID- 16379394 TI - Selenium species determination in selenium-enriched pumpkin (Cucurbita pepo L.) seeds by HPLC-UV-HG-AFS. AB - Pumpkins were treated by spraying the leaves in the flowering period with a water solution containing 1.5 mg Se per liter in the form of Na2SeO4. The average total selenium content of seeds was found to be 0.19 microg g(-1) in nontreated pumpkins and 1.1 microg g(-1) in exposed ones. For speciation analysis, enzymatic hydrolysis with different amounts of Protease XIV was carried out. Under optimal conditions of enzymatic hydrolysis, 90% of the total selenium was found in soluble forms. Separation of species was performed using HPLC on anion and cation exchange columns and for detection UVHG-AFS was applied. In enzymatic hydrolysis extracts, the main fraction of selenium was bound as selenomethionine (SeMet), representing on average of 81 +/- 8% of the total Se content in the sample. PMID- 16379395 TI - Rapid extraction of water soluble organic compounds from airborne particulate matter. AB - Water soluble organic compounds (WSOC) in airborne particulate matter (PM) have received considerable attention in recent years due to their abundance and their importance in atmospheric processes. The analysis of WSOC is necessary for quantifying the relative contribution of individual organic compounds to the total WSOC mass. In the present work, we evaluated the performance of a microwave assisted extraction (MAE) method for the determination of WSOC in PM and compared the data with those of a conventional ultrasonic extraction (USE). The experimental results showed that the MAE method requires a shorter extraction time (5 min) compared to USE. The isolated water-soluble organic fraction of PM was subsequently analyzed using ion chromatography (IC) for low molecular weight organic acids. The rapid MAE method was used in conjunction with IC for the analysis of organic acids in PM samples, collected from different sources. PMID- 16379396 TI - Application of sequential injection analysis to the determination of cationic surfactants based on the sensitized molybdenum-bromopyrogallol red reaction. AB - The aim of this work was to develop a simple, automatic system for the evaluation of cationic surfactants by combining sequential injection analysis and the sensitized effect of cationic surfactants on the reaction between metal ions and chelating dyes. This reaction is based on the increase in absorbance of the complex formed among molybdenum, bromopyrogallol red and increasing concentrations of cationic surfactants. Under optimum conditions, two calibration plots were obtained for a concentration range between 2.50 x 10(-7) mol dm(-3) (detection limit) and 5.00 x 10(-4) mol dm(-3) of cetylpyridinium chloride, used as standard. Solubilization of water insoluble complexes formed for concentrations of cationic surfactants greater than 1.00 x 10(-4) mol dm(-3) were successfully achieved with Triton X-405. RSD values lower than 5.0% were obtained in all cases. The quality of the results obtained for 18 water samples were evaluated by comparison with conventional methods, with no statistically significant differences for a 95% confidence level. PMID- 16379397 TI - Development and validation of a flow-injection assay for dissolution studies of the anti-depressant drug venlafaxine. AB - The first flow-injection method has been developed, optimized and validated for the determination of venlafaxine, an antidepressant drug. The method is based on a direct measurement of the absorbance of the analyte in an acidic medium, at 274 nm. Flow-injection parameters, such as sample injection volume and flow rate, were studied and optimized. The proposed method was validated in terms of linearity, repeatability, detection limit, accuracy and selectivity. Linearity was obeyed in the range 30 - 150 mg L(-1) of venlafaxine, while the detection limit (1.5 mg L(-1)) and repeatability (sr < 1.0%, n = 12) were satisfactory. The sampling rate was 30 h(-1). The results of dissolution studies of venlafaxine tablets obtained by the proposed method were in good agreement with those by high performance liquid chromatography. PMID- 16379398 TI - Determination of chromium, copper and lead in river water by graphite-furnace atomic absorption spectrometry after coprecipitation with terbium hydroxide. AB - Coprecipitation with terbium hydroxide quantitatively recovered trace amounts of chromium(III), copper(II) and lead(II) at pH 8.4 - 10.8, 8.0 - 11.5 and 8.7 - 11.5, respectively. The precipitate was dissolved in 0.85 mol dm(-3) nitric acid, and the analytes were determined by graphite-furnace atomic absorption spectrometry (GF-AAS). The presence of terbium (up to 7 g dm(-3)) did not interfere with the determination. The detection limits were 0.3 microg dm(-3) for chromium, 0.4 microg dm(-3) for copper and 0.5 microg dm(-3) for lead, when the analytes in 200 cm3 of the sample solution were concentrated into 10 cm3. The ions added to river or seawater were quantitatively recovered. Chromium and copper in a contaminated river water were successfully determined. PMID- 16379399 TI - Exploratory analysis of simultaneous degradation of anthocyanins in the calyces of flowers of the Hibiscus sabdariffa species by PARAFAC model. AB - A three-way resolution method based on PARAFAC model was applied for the UV-Vis spectra to study the simultaneous degradation of anthocyanins extracted from fresh calyces of flowers of the Hibiscus sabdariffa. This methodology was used to resolve a chemical system, for which there was no a priori information about the composition or the pure spectra, a so-called black system. In the pH range from 1 to 13, seven species were identified: flavylium cation, carbinol, quinoidal base, E- and Z-chalcones and E- and Z-ionized chalcones. The concentration changes were determined as functions of pH at different wavelengths. The pK values for the acidity constants as well as tautomeric constant were estimated as 2.70, 3.54 and 0.14, respectively. The spectral profiles recovered by the chemometric methods are in excellent agreement with bands of experimental spectra reported in the literature for the species measured at specific pH values. PMID- 16379400 TI - Voltammetric behavior of the transfer of mono- and polyammonium ions across a phospholipid monolayer at the nitrobenzene/water interface. AB - The influence of a phospholipid, dipalmitoyl phosphatidylcholine, layer at a nitrobenzenelwater interface on the transfer of tetraethylammonium ion and a polyammonium anti-fungus agent, poly[(dimethylimino)(2-oxo-1,2 ethanediyl)imino1,6-hexanediylimino (1-oxo-1,2-ethanediyl)(dimethylimino)-1,6 hexanediyl] ion, across the interface was studied by normal pulse voltammetry. When the phospholipid was adsorbed to form a monolayer at the nitrobenzenelwater interface by its addition to the organic phase, the half-wave potential in the current vs. potential curves for the transfer of tetraethylammonium ion did not change, but the limiting current was significantly decreased at certain sampling times, indicating a retarding effect of the layer on the ion-transfer. On the other hand, in the current vs. potential curves for the transfer of the polyammonium ion, no significant change in either the half-wave potential or the limiting current was observed upon adding the phospholipid, indicating that the polyammonium ion can easily permeate through the phospholipid layer. The results suggest a new application of the voltammetric technique to the study of cell membrane permeability to polyionic bioactive compounds. PMID- 16379401 TI - Preparation of a novel iodide-selective electrode based on iodide-miconazole ion pair and its application to pharmaceutical analysis. AB - An iodide-miconazole ion-paired complex was used as a suitable ion-exchanger for the preparation of a plasticized-PVC membrane electrode. Among different solvent mediators tested, dioctylsebacate exhibited the proper response characteristics, including Nernstian slope of the calibration curve, fast response time and good reproducibility of the emf values. The electrode exhibits a Nernstian slope of 59.8 +/- 0.5 mV decade(-1) for I- ion over a concentration range of 1.0 x 10(-5) 1.0 x 10(-2) M with a limit of detection of 7.0 x 10(-6) M. The electrode displays a good selectivity for I- with respect to a number of inorganic and organic species. It canbe used over a pH range of 2.5 - 8.5. The membrane sensor was successfully applied to the determination of iodide in water samples and blood serum, as well as in pharmaceutical products such as iodoquinol and thyroxin. PMID- 16379402 TI - Ultrafast diagnosis of the genetic-related disorders using the combined technologies of multiplex PCR and multichannel microchip electrophoresis. AB - For the diagnosis of unexplained male infertility a multiplex PCR for 6 markers, which are well-known as candidate genes for studying male infertility and located on the human Y-chromosome, has been designed. The multiplex PCR products have been separated on a 12 channel microchip electrophoresis system, which can analyze different samples simultaneously. By combining the technologies of multiplex PCR with multichannel microchip electrophoresis, the number of the DNA markers that can be screened simultaneously is increased to be 72 marker (12 x 6) in a single run while the electrophoresis analysis time is reduced to be only 180 s. PMID- 16379403 TI - Uptake of cycloalkanes and bicyclic aromatic compounds by ion-exchange resins from aqueous solutions. AB - The uptakes of cycloalkanes (C5 - C8) and bicyclic aromatic compounds (naphthalene, biphenyl, and azulene) by both strong acid cation (Dowex 50W-X4 and X8) and strong base anion (Dowex 1-X4 and X8) exchange resins have been studied in aqueous solutions at 25 degrees C; their distribution constants (K) have been determined. The relationship between the octanol-water partition coefficient and the resin affinity for solutes has been analyzed. The K values of three aromatic compounds were found to be much higher than expected from the octanol-water partition coefficients. This is probably due to the pi interaction of aromatic rings with the resin matrix. Microscopic observations revealed that azulene molecules are uniformly distributed inside a resin particle. PMID- 16379404 TI - Simultaneous determination of tocopherols and tocotrienols in hazelnuts by a normal phase liquid chromatographic method. AB - A normal-phase high-performance liquid chromatography (NP-HPLC) method for the determination of tocopherols and tocotrienols in hazelnuts is reported. Three extraction procedures (with and without saponification) were assayed; the best results were obtained with a simple solid-liquid extraction procedure. Chromatographic separation was achieved using an Inertsil 5 SI column using isocratic elution with hexane/1,4-dioxane (95.5:4.5, v/v) at a flow rate of 0.7 mL/min. The effluent was monitored by a series arrangement of a diode-array followed by a fluorescence detector. All compounds were separated in a short period of time (17 min). The method proved to be rapid, sensitive, reproducible and accurate, allowing the simultaneous determination of all vitamin E homologues. PMID- 16379405 TI - Spectrophotometric determination of aluminum with m-carboxyphenylfluorone, a novel chemical probe, and its application. AB - A highly spectrophotometric method for the determination of aluminum was developed. This method used the color reaction between m-carboxyphenylfluorone (MCPF) as a novel chemical probe and aluminum in the presence of a surfactant, poly(N-vinylpyrrolidone) (PVP, K-90) (0.03 - 1.40 microg of aluminum in a final volume of 10 ml at 561 nm). The proposed method showed excellent sensitivity (molar absorptibity of 1.70 x 10(5) l mol(-1) cm(-1)) and reproducibility (within day precision: RSD = 0.35% n = 6, between-day precision: RSD = 0.44% n = 6). Linearity was achieved over the range 3 - 140 microg L(-1) with a correlation coefficient of 0.9999, and the effects of foreign substances were low. PMID- 16379406 TI - Determination of Cu, Pb, Fe, and Zn in plant component polymers of a hyperaccumulating plant. AB - Phytoremediation is an innovative technology that utilizes the natural properties of plants to remediate hazardous waste sites. For more cost-effective phytoremediation, it is important to utilize a hyperaccumulating plant after phytoremediation, i.e. the recovery of valuable metals and the production of useful materials. In this work, the determination of metals in plant component polymers in a fern, Athyrium yokoscense, as a hyper-accumulating plant was established using steam explosion, Wayman's extraction method, and ICP emission spectrometry. After A. yokoscense plants were treated by steam explosion, the steam-exploded A. yokoscense were separated into four plant component polymers, ie. water-soluble material fraction, holocellulose fraction, methanol-soluble lignin fraction, and residual lignin fraction. The concentrations of Cu, Pb, Fe, and Zn in these plant component polymers and the dry weights of plant component polymers were measured. These analytical process determining metals in the plants will contribute to not only the evaluation and the efforts of phytoremediation using a hyperaccumulating plant, but also to the development of more effective phytoremediation. PMID- 16379407 TI - Simultaneous spectrophotometric analysis of aliphatic amines utilizing thermochromism of charge-transfer complexes with tetrabromophenolphthalein ethyl ester. AB - Aliphatic amines, such as n-hexylamine (primary), di-n-hexylamine (secondary) and tri-n-hexylamine (tertiary amine), react with tetrabromophenolphthalein ethyl ester molecules (TBPEH) to form reddish or red-violet charge-transfer complexes (CT complexes) in 1,2-dichloroethane (DCE). The absorption maxima of the CT complexes with all primary amines occur at around 560 nm, with secondary amines at 570 nm and, with tertiary amines at 580 nm. The CT complex formation constants with TBPEH in DCE increase in the order of the primary, secondary and tertiary amines, but their constants decrease quantitatively with an increase in temperature. This phenomenon (thermochromism) could be applied to the simultaneous spectrophotometric determination of primary amine and secondary amine, or secondary amine and tertiary amine in a mixed solution utilizing the difference of absorbance with temperature changes. PMID- 16379408 TI - A new objective method to compare different color reactions for the detection of cyclic ureides. AB - Modifications of the Zwikker- and Parri color detection tests were examined and compared according to their ability to distinguish between nine different barbituric acids and hydantoins. These tests comprised the formation of complexes with cobalt(II) and copper(II) salts and organic amines. Using the color palette introduced herein, the evaluation of the tests could be reduced to a simple yes/no decision on the basis of only seven defined colors. Suitable components for the color tests could be selected from the thirty six modifications. PMID- 16379409 TI - Model-based evaluation of diabetic foot prevention strategies in Austria. AB - Diabetes mellitus affects approximately 171 million individuals worldwide. The costs of the adult form of diabetic mellitus account for up to 6% of total health care expenditures in industrialized countries. About 25% of these diabetics develop disabling and most painful foot complications accounting for about 17% of the direct lifetime costs. Diabetic foot prevention programs have been recently introduced in some Austrian federal states to meet the diabetic health targets of the Austrian Health Plan and the St. Vincent Declaration. We developed a new age group specific Markov model combined with a Monte Carlo simulation model to help policymakers analyze the cost-effectiveness of such programs compared to the status quo in terms of incremental costs per quality-adjusted life years gained (QALY). The Markov model revealed that diabetic foot prevention programs were cost saving when targeted at patients at high risk and mainly cost-effective when targeted at patients with mild symptoms. The Monte Carlo simulation showed that only large scope prevention programs would fulfill the specified reductions in the number of diabetic foot complications as defined in the Austrian Health Plan and the St. Vincent Declaration. Our results clearly indicate the enormous impact of diabetic foot prevention programs. PMID- 16379410 TI - Analysis and implications of the determinants of healthcare expenditure in African countries. AB - The income elasticity of health care spending in the OECD countries tends toward luxury good values. Similar studies, based on more recent data, and capable of informing macroeconomic health policies of the African countries, do not currently exist. How the health care expenditure in Africa responds to changes in the Gross Domestic Products (GDP), Official Development Assistance (ODA), and other determinants, is also relevant for health policy because health care is a necessity in the 'basic needs' theory of economic development. This paper presents econometric model findings of the determinants of per-capita health expenditure (in PPPs) for 26 African countries, using the flexible Box-Cox model regression methods and 1995 cross-sectional data (sources: WRI, UNEP, UNDP, The World Bank). The economic and other determinants, capturing 74 percent of the variations in health expenditures, include per-capita GDP (in PPPs), ODA (US dollar), Gini income inequality index, population dependency ratio, internal conflicts, and the percentage of births attended by trained medical workers. Income inequality dampens, while the ODA and population per health personnel raise health care expenditure. The GDP elasticity of about 0.6 signals the tendency for health care to behave like a technical 'necessity'. Implications for sustainable basic health development policies are discussed. PMID- 16379411 TI - The effect of social mixing controls on the spread of smallpox--a two-level model. AB - Responding to a possible bioterror attack of Smallpox has become a major concern to governments, local public officials and health authorities. This concern has been reflected in numerous studies that model and evaluate possible response strategies. Many of these studies consider only vaccination policies and assume homogeneous mixing, where all instances of contacts in the population are equally likely. Such a mixing pattern is rather unlikely to represent population interaction in a modern urban setting, which typically is separated into households on the one hand, and into daily meeting sites such as schools and offices, on the other hand. In this paper we develop a two-level social interaction model where an individual moves back and forth between home and a daily meeting site, possibly passing through a general meeting site such as mass transit system or other crowded areas. Based on the model, we evaluate the effect of social mixing controls, situational awareness of the public health system and mass vaccination on the spread of smallpox. It is shown that mixing controls and alertness of the response system may have a significant impact on the spread of the epidemic. Some policy recommendations are discussed. PMID- 16379412 TI - Methods for understanding super-efficient data envelopment analysis results with an application to hospital inpatient surgery. AB - We compare two techniques for increasing the transparency and face validity of Data Envelopment Analysis (DEA) results for managers at a single decision-making unit: multifactor efficiency (MFE) and non-radial super-efficiency (NRSE). Both methods incorporate the slack values from the super-efficient DEA model to provide a more robust performance measure than radial super-efficiency scores. MFE and NRSE are equivalent for unique optimal solutions and a single output. MFE incorporates the slack values from multiple output variables, whereas NRSE does not. MFE can be more transparent to managers since it involves no additional optimization steps beyond the DEA, whereas NRSE requires several. We compare results for operating room managers at an Iowa hospital evaluating its growth potential for multiple surgical specialties. In addition, we address the problem of upward bias of the slack values of the super-efficient DEA model. PMID- 16379413 TI - Transitions between care provisions demanded by Dutch elderly. AB - We follow people who are considered eligible for an elderly care provision. Through the estimation of a duration model we determine when they can be expected to express demand for another care provision. Data are drawn from the registrations of needs assessment agencies in The Netherlands. MAIN FINDINGS: Age positively effects the probability of a reassessment if initially care at home was assessed. Living alone, access to informal care, and applications initiated by professionals increase transition probabilities. Also initial applications for care at home due to dementia are more likely to be followed by a quick reassessment. The results can aid assessment agencies to set validity terms of assessments, and in planning future supply of services. PMID- 16379414 TI - Short-term nurse scheduling in response to daily fluctuations in supply and demand. AB - Hourly changes in patient census and acuity require hospitals to update their staffling needs on a continuing basis. This paper discusses the problem that management faces several times a day as the demand for nursing services departs from the planned schedule. Prior to the start of each shift, the number of nurses who are scheduled to be on duty over the next 24 hours is compared with the number actually available, and if shortages exist a series of decisions have to be made to ensure that each unit in the hospital has sufficient coverage. These decisions involve the use of overtime, outside nurses, and floaters. To address this problem, we have developed an integer programming model that takes the current set of rosters for regular and pool nurses and the expected demand for the upcoming 24 hours as input, and produces a revised schedule that makes the most efficient use of the available resources. The model is formulated and solved at a hospital-wide level rather than for each unit separately. To determine its applicability, a representative set of scenarios was investigated using data obtained from a medium-size facility in the U.S. with 14 units. The results indicate that problem instances with up to 120 nurses can be solved in a negligible amount of time. PMID- 16379415 TI - Modelling variability in hospital bed occupancy. AB - A stochastic version of the Harrison-Millard multistage model of the flow of patients through a hospital division is developed in order to model correctly not only the average but also the variability in occupancy levels, since it is the variability that makes planning difficult and high percent occupancy levels increase the risk of frequent overflows. The model is fit to one year of data from the medical division of an acute care hospital in Adelaide, Australia. Admissions can be modeled as a Poisson process with rates varying by day of the week and by season. Methods are developed to use the entire annual occupancy profile to estimate transition rate parameters when admission rates are not constant and to estimate rate parameters that vary by day of the week and by season, which are necessary for the model variability to be as large as in the data. The final model matches well the mean, standard deviation and autocorrelation function of the occupancy data and also six months of data not used to estimate the parameters. Repeated simulations are used to construct percentiles of the daily occupancy distributions and thus identify ranges of normal fluctuations and those that are substantive deviations from the past, and also to investigate the trade-offs between frequency of overflows and the percent occupancy for both fixed and flexible bed allocations. Larger divisions can achieve more efficient occupancy levels than smaller ones with the same frequency of overflows. Seasonal variations are more significant than day-of-the-week variations and variable discharge rates are more significant than variable admission rates in contributing to overflows. PMID- 16379416 TI - Determinants of dental service utilization among adults--the case of Finland. AB - We examine the determinants of utilization of dentists' services and especially the role of supply factors in the decision-making processes of utilization, using data from the Finnish Health Care Survey of 1996. In a two-part model, care seeking is modeled by a bivariate probit model taking into account dentist's recall, whereas the number of positive visits to the dentist is modeled by a zero truncated negative binomial model. Recall turns out to be a central determinant of care-seeking and is significantly related to variables like income and unemployment. Socioeconomic variables are controlled for as well and are found to be important determinants. PMID- 16379417 TI - [Study on dynamics of tannin in different growth phase of Cynomorium songaricum]. AB - Potassium permanganate titration and complexometry were used to determine the content of tannin in different growth phase of wild and cultivated Cynomorium songaricum Rupr. The result showed that the content appeared regularly changing and the content in coming-up phase was higher than other phases. The quality in this phase would be better if the tannin was regarded as one of the effective components. The result might provide science foundation for the quality control of Herb Cynomorii. PMID- 16379418 TI - Studies on chemical constituents of Rheum glabricaule. AB - OBJECTIVE: To study the chemical Constituents in the root of Rheum glabricaule. METHODS: Compounds were isolated by various column chromatographies with sillica gel. Their structures were identified by spectral analysis (MS, 1HNMR, 13 CNMR) and chemical evidences. RESULTS: Seven compounds were isolated from this plant, including n-hexacosnic acid (I), palmitic acid (II), daucosterol (III), chrysophanol-8-Me ether (IV), citreorosein (V), chrysophanol 8-O-beta-D glucopyranoside (VI) and 2,5-dimethyl-7-methoxychromone (VII). CONCLUSION: All above compounds are obtained from this plant for the first time. PMID- 16379419 TI - [Study on the effect and its mechanisms of 5F from Pteri semipinnata L. on the cell cycle of highly metastatic ovarian carcinoma HO-8910PM cells]. AB - OBJECTIVE: To study the effect and its possible mechanism of 5F from Pteri semipinnata L on the cell cycle of human highly metastatic ovarian carcinoma HO 8910PM cells. METHODS: MTT assay was used to examine the effect of 5F on proliferation of HO-8910PM cells after 24 hours treatment; The cell cycle was assessed by flow cytometry (FCM); The expression level of NF-kappaB (p65) FAK and the level of phosphorylated FAK were assessed by Western blot analysis. RESULTS: 5F from Pteris semipinnata L. could inhibit the proliferation of HO-8910PM cell and block the cell cycle at G2/M phase. The expression level of NF-kappaB (P65) protein decreased obviously in HO-8910PM cells treated with 25 - 100 micromol/L 5F for 24 hours, and the effect appeared in a dose-dependent manner. 5F up regulated significantly the expression of FAK, down-regulated the level of phosphorylated FAK. CONCLUSION: The effect on cell cycle of 5F from Pteri semipinnata L is involved in the expression of NF-kappaF(p65), FAK and the level of phosphorylated FAK. PMID- 16379420 TI - [Effect of Laminaria japonica polysaccharides on lowing serum lipid and anti atherosclerosis in hyperlipemia quails]. AB - OBJECTIVE: To study the effect of Laminaria Japonica Polysaccharides on lowing serum lipid and anti-atherosclerosis in quails. METHODS: The hyperlipemia atherosclerosis (As) models of quails were induced by high lipids. After administration with Laminaria Japonica Polysccharides for two weeks, the content of total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL) and low density lipoprotein (LDL) in serum and the ratio of HDL/TC were determined, index of liver was calculated, the extent of aortic atherosclerosis and the atherosclerotic lesion were observed. RESULTS: The concentration of TC, TG, LDL in serum and index of liver decreased remarkedly, the ratio of HLD/TC increased remarkably, the the extent of aortic atherosclerosis and the atherosclerotic lesion lightened obviously. CONCLUSION: Laminaria Japonica Polysaccharides can effectively decrease the serum lipid and prevent the formation of atherosclerosis. PMID- 16379421 TI - [Experimental study on anti-DHBV effect of yu gan capsules]. AB - OBJECTIVE: To observe the antiviral effect of Yu Gan capsules against DHBV in ducks. METHODS: 50 Guangzhou brown spot ducks were used in this experiment. After the ducks were infected DHBV, they were divided into four groups at random (ACV 200 mg/kg group, Yu Gan capsules 6.5 g/kg group, Yu Gan capsules 3.25 g/kg group and virus control group) and Drugs were given by stomach once a day. Blood samples were taken from each duck and tested for DHBV-DNA's level. Researches left liver tissue were, sheared and used for optical analysis. RESULTS: In Yu Gan capsules 6.5 g/kg group, DHBV-DNA's level in ducks' serum decreased on the 5th and 10th days after treatment and the 3th day after recession of treatment. There was a significant difference in the DHBV-DNA's level between the days after treatment and before treatment. there was also significant difference in the DHBV DNA's level between Yu Gan capsules 6. 5 g/kg group and virus control group (P <0.01). Whereas, no inhibition effect was found in Yu Gan capsules 3.25 g/kg group. CONCLUSION: Yu Gan capsules have functions on anti-DHBV in duck. PMID- 16379422 TI - [Study on the vasodilation effect of blood vessel and its mechanism by the extract of bu shen huo xue recipe]. AB - OBJECTIVE: To study the effect of the extract of Bu Shen Huo Xue Recipe on blood vessel and the content of NO, NOS, cNOS and iNOS in plasma and aortic tissue. METHODS: The effect of blood vessels was studied by guinea pig mesentery blood capillary, rat aortic ring and celiac vein ring. The content of NO, NOS, cNOS and iNOS were determined by method of nitroreductase chromatometry in plasma and aortic tissue. RESULTS: The extract of Bu Shen Huo Xue Recipe could significantly increase the sum of crisscross blood capillary, improve bloodstream state and speed. The significantly vasodilating effect induced by the extract of Bu Shen Huo Xue Recipe was seen in rat aortic ring and celiac vein (based on 10 micromol/L phenylephrine-induced concentration). But there was contracting effect in denuded endothelium. The relaxations induced by the extract on blood vessels mentioned above were markedly attenuated by preincubation in organ chambers with glibenclamide (1 micromol/L), the same effect in lower dose with propranolol (3 micromol/L), the contractional effect in high dose with propranolol (3 micromol/L). The extract in high dose could significantly affect the content of NO in plasma, increase the content of NO, NOS and cNOS in aortic tissue. CONCLUSION: The mechanism of vasodilation effect of blood vessel of the extract may be related to releasing NO from endothelium of blood vessels and increasing the content of synthetic NOS and cNOS in aortic tissue. PMID- 16379423 TI - [Study on pharmacodynamics of run chang tong bian granules]. AB - OBJECTIVE: To study the effect of Run Chang Tong Bian Granules (RCTBG) on stomach and intestine peristalsis in mice and rabbits. METHODS: We conducted the intestine peristalsis test and gastric emptying test to observe the purgative action of RCTBG and the effect on isolated intestine of rabbits. RESULTS: RCTBG could effectively expedite the advance speed of charcoal powder in the small intestine and improve the velocity of intestine. It could also reduce the residual rate of methyl orange in stomach and improve the velocity of gastric emptying. RCTBG could remarkably shorten the time of defecation, increase excrement quantity of mice, excite the isolated intestine of rabbits and increase its tension and contraction amplitude. CONCLUSION: RCTBG has remarkable effect on catharsis. PMID- 16379424 TI - [Study on the effect of promoting intelligence development and preventing hypoxia/reoxygenation injury of selenium-banqiao-Codonopsis pilosula-overground part in mice]. AB - OBJECTIVE: To study on the effect of promoting intelligence development and preventing Hypoxia/Reoxygenation injury of Selenium-Banqiao-Codonopsis pilosula overground part in mice. METHODS: Promoting Intelligence Development experiment was induced by PIA; Hypoxia/reoxygenation ingury model was established to observe the activity of ROS, SOD, MOD and CAT in blood. RESULTS: Selenium-Banqiao Codonopsis pilosula-overground part could enhance the learning and memory ability of old mice and obviously extend the swimming time of mice. It could also decrease the quality of ROS and MDA, increase the activity of SOD, but no significant effect on CAT. CONCLUSION: Selenium-Banqiao-Codonopsis pilosula overground part has effect on promoting intelligence development and preventing hypoxia/reoxygenation injury. PMID- 16379425 TI - [Study on the effect of Ginkgo biloba extract on the tension of diabetic rat artery]. AB - OBJECTIVE: To investigate the effect and possible mechanism of Ginkgo biloba extract EGB50 on vascular tension of type II diabetes mellitus (DM) induced by hyperglycemia and hyperlipidemia. METHODS: Rats were randomly divided into normal control rats (Control), diabetic rats (DM), diabetic rats oral-treated with higher dose Ginkgo biloba extract EGB50 (H) and diabetic rats treated with lower dose EGB50 (L). The serum levels of Advanced Glycosylation end-products (AGEs), cholesterol (CHOL), triglyceride (TRIG) and superior mesenteric artery tension were quantified and measured. RESULTS: After 5 weeks' oral-treatment, serum CHOL, TRIG and AGEs increased, sustained phase of contractile response in high K+ solution and PD2 of phenylephine decreased in diabetic arteries. The biochemical indexes and the tension of vascular function had less significance in L group, but improved distinctly in EGB50 H group (decreasing rate in high K+ solution: 22.52 +/- 5.48%, vs DM:44. 19 +/- 11.03%, P < 0.05) (PD2 of PE: 6.15 +/- 0.22 vs DM: 6.62 +/- 0.13, P < 0.05). CONCLUSION: EGB50 is capable of reducing serum hyperlipidemia,the concentration of AGEs, thus it can reduce the impairment of the oxidants to endothelium cells of vessels and improve pathological and functional change of artery of diabetes. PMID- 16379426 TI - [Study on the extraction and purification for total flavonoids in Lonicera japonica Thunb. and Chrysanthemum morifolium Tzvel]. AB - OBJECTIVE: To optimize the extraction and purification conditions for total flavonoids in Lonicera japonica Thunb. and Chrysanthemum morifolium Tzvel. METHODS: Comparision of several extraction processes, the L9 (3(4)) orthogonal design and macroreticular absorption resin D101 purification technique were performed. RESULTS: The optimum extraction conditions were 8 volume of 55% ethanol, 1 h refluence at 100 degrees C , water bath for 2 times. The extracted fluid was absorbed with the resin D101. The elutions were water, 30% enthanol, 70% enthanol. The obtained elution of 70% enthanol was evaporated and the content of total flavonoids was 62. 7%. CONCLUSION: The method is simple, approved and fit for industrial production. PMID- 16379427 TI - [Study on new extraction technology of astragaloside IV]. AB - OBJECTIVE: To explore the possibility and the optimal extraction technology of astragaloside IV by SFE-CO2. METHODS: According the content of astragaloside IV, the optimum extraction technology parameters such as extraction temperature, pressure, extraction time, velocity of fluid and co-solvent were investigated and the result was compared with that of water extraction. RESULTS: The optimum technical parameters were as follows: Extracting pressure 40 Mpa, temperature 45 degrees C, extracting time 2h, co-solvent was 95% ethanol and its dosage was 4ml/g, the ratio of CO2 fluid was 10 kg/kg x h. CONCLUSION: Extraction technology of astragaloside IV by SFE-CO2 is reliable, stable. PMID- 16379428 TI - [Study on preparation technological process of huangqin qingfei koufuye]. AB - OBJECTIVE: To study the optimum preparation technological process of Huangqin Qingfei Koufuye for mass production. METHODS: The optimum technology of extraction HCN from Semen Armeniacae Amarum was selected by experiments; The optimum conditions for the extraction process was selected with orthogonal design, and the content of baicalin in the extract as marker was determined. RESULTS: The optimum technology of extraction HCN from Semen Armeniacae Amarum was as follows: crushing to pieces, adding 3 times amount of 37 degrees C water and soaking overnight, extracting and collecting 1.33 times amount of extract; the optimum extraction process wasas follows: extracting 2 times, each time adding 12 times amount of water and extracting 1 h. CONCLUSION: The preparation technological process can be used for mass production. PMID- 16379429 TI - [Clinical study of matrine injection on preventing liver function damage of anti tumor drugs during chemotherapy of breast cancer]. AB - OBJECTIVE: To study the effect of matrine injection on preventing liver function damage of anti-tumor drugs during chemotherapy of breast cancer. METHODS: 167 times of chemotherapy were performed and divided into two groups (the control and treatment group: 86 times for the treatment one, 81 times for the control). In control group, regular hepatinica was used, conbining with matrine injection 150 mg and infusion once a day for two weeks in treatment group. Serum levels of alamine aminotransterase (ALT), aspartate amiotransferase (AST), gamma-glutamyl transferase (GGT), Serum total bilirubin (STB) were tested before, after two weeks and after three weeks of chemotherapy in two groups. RESULTS: The levels of ALT, AST, GGT, STB significantly increased (P < 0.05). The levels of ALT and AST in control group were higher than that in treatment one. There were no severe side-effects of matrine. CONCLUSION: Matrine injection may be used safely and possess less side-effects on lowing serum levels of ALT, AST, GGT, STB in patients with breast cancer during chemotherapy so it can be used as adjuvant to prevent damages of the liver function for chemotherapy of patients with breast cancer. PMID- 16379430 TI - [Clinical study on "sini" decoction on treating stenocardia for coronary heart disease]. AB - OBJECTIVE: To observe clinical effect of "Sini" decoction on stenocardia for coronary heart disease. METHODS: 65 cases of stenocardia for CHD with deficiency of yang or cold syndrome were randomly divided into" Sini" decoction therapeutic group and isosorbide dinitrate controll group. Drug effects of the two groups were compared by clinical symptoms, electrocardiogram (ECG), myocardial oxygen consumption and cardiac function. RESULTS: "Sini" decoction had better effects on reducing myocardial oxygen consumption and improving cardiac function isosorbide dimitrate, and similar to improving clinical symptoms and ECG, decreasing frequency of stenocardia and dose of nitroglycerin on stenocardia for coronary heart disease. CONCLUSION: "Sini" decoction can treat stenocardia for CHD with deficiency of yang or cold syndrome. PMID- 16379431 TI - Trends in indirect dentistry: 8. CAD/CAM technology. AB - Computer-aided design (CAD) and computer-aided manufacturing (CAM) of indirect restorations became available to dentistry over 15 years ago, providing replication and digitization of the complex topography of tooth structure. There are now many applications, providing better mechanical properties, improved marginal integrity and enhanced aesthetics, compared with traditional indirect techniques. Whether a restoration is fabricated by traditional or modern computerized systems, three functional stages are required; data acquisition, design and manufacture. It is the purpose of this paper to describe the applications of CAD and CAM in contemporary indirect dentistry. CLINICAL RELEVANCE: Reliable CAD/CAM techniques, some of which may reduce the number of patient visits, are now available for the production of a variety of ceramic restorations. PMID- 16379432 TI - Trends in Indirect Dentistry: conclusions. PMID- 16379433 TI - Oral health needs of the head and neck radiotherapy patient: 2. Oral and dental care before, during and after radiotherapy. AB - Head and neck cancer represents a heterogeneous group of neoplasms affecting a number of sites in the aerodigestive tract, histologically primarily epithelial in type. Most are sensitive to a treatment regime of ablative surgery followed by radiotherapy. The incidence of head and neck cancers is approximately 11.8 per 100,000 population and is the 8th commonest cancer in males and the 16th in females. Treatment comprises ablative surgery with, in 60% of cases, radiotherapy. The oral and dental sequelae of such treatment, and their management, is described. CLINICAL RELEVANCE: Many of these cancers will manifest with oral signs and symptoms, thus placing the GDP in the unique position of being able to detect and positively influence outcomes. As well, the GDP has a vital, proactive role to play in supportive care before, during and following surgery and radiotherapy. PMID- 16379434 TI - Alloys for metal-ceramic restorations. AB - The requirements of a metal-ceramic dental restoration consisting of a rigid substrate, which supports an aesthetic veneer, are assessed. The contemporary, cast, bonding alloys that have good track records as substrates are categorized and their properties are compared. Several alternatives to cast metals are considered. CLINICAL RELEVANCE: Despite developments in all-ceramic restoration technology, the metal-ceramic dental restoration still has a significant place in the dentist's armamentarium. PMID- 16379435 TI - Bitewing examination to detect caries in children and adolescents--when and how often? AB - It is generally agreed that the decision to take bitewing radiographs for detecting caries should be based on the benefit to the individual patient in relation to the risks associated with low dose radiation exposure and the costs. There is incomplete knowledge about the effectiveness of various methods for selecting individuals who will benefit from bitewing examination. Available knowledge suggests, however, that our ability to identify correctly those who will benefit is limited. It may, therefore, be more effective to combine population- and individual-based selection criteria. For this purpose, four key ages and individual-based criteria between the key ages are suggested. CLINICAL RELEVANCE: The purpose of the presented selection criteria are to improve the dental practioner's effectiveness in selecting patients who will benefit from bitewing examination, thereby reducing the number of arbitrary and 'just in case' radiographs and the radiation dose. PMID- 16379436 TI - Periodontal disease and the ageing patient. AB - The UK's population is getting older and the number of older individuals retaining their teeth is increasing. Greater demands are therefore being placed on general dental practitioners for periodontal therapy. Many factors, including the ageing process itself, impact directly or indirectly on periodontal health. This article discusses these factors, their interrelationship and their relevance to clinical management of periodontal disease in older patients. CLINICAL RELEVANCE: This article aims to give the practitioner an insight into the issues involved in managing older patients with periodontal diseases. PMID- 16379437 TI - Overeruption--another challenge? AB - Awareness of the occlusal changes following individual tooth loss may be increasingly important when planning treatment for patients who are likely to retain a relatively intact dentition throughout life. Some evidence exists to support dentists who are considering prevention or treatment of overeruption. This paper explores the literature available on the changes in the prevalence of tooth loss, the prevalence and presentation of overeruption, its relevance to dental restoration and the prevention and treatment of overeruption. CLINICAL RELEVANCE: This article reviews the prevalence and consequence of overeruption in posterior teeth. PMID- 16379438 TI - The consistent image--how to improve the quality of dental radiographs: 1. Quality scale, operator technique, X-ray set. AB - There are two papers in this series aiming to put into context quality assurance as applied to dental radiography. 'The Ionising Radiation Regulations' emphasize the need for quality assurance programmes wherever radiography is being undertaken. A quality assurance programme should aim to be written down common sense. This first paper deals with assessment of image quality, faults arising from poor operator technique and exposure factors. CLINICAL RELEVANCE: The object of a quality assurance programme is to minimize the dose to the patient whilst maximizing diagnostic utility. PMID- 16379439 TI - An unusual case of floor of mouth swelling: case report, differential diagnosis and a review of the literature. AB - The oral cavity is an uncommon site for a true lipoma. A distinct histological variant is the spindle cell lipoma. This is a rare oral lesion. A case of an intra-oral spindle cell lipoma in a 55-year-old female is reported, a differential diagnosis is presented and the histopathology is discussed. CLINICAL RELEVANCE: An oral swelling is a common presenting complaint and requires further investigation. PMID- 16379440 TI - Physical signs for the general dental practitioner. PMID- 16379441 TI - Nine years to end an injustice. PMID- 16379442 TI - Like it or not, nurse prescribing is here to stay. PMID- 16379443 TI - Night nursing. PMID- 16379445 TI - Lymphoma. PMID- 16379444 TI - Use and practice implications of modified-release medicines. AB - Modified-release drugs have complex formulations and require extra care to ensure safe prescribing and administration. This article discusses the use of modified release preparations and highlights their practice implications. PMID- 16379446 TI - The Liver. Part 3: Investigations of liver function. PMID- 16379447 TI - The evidence base for using preoperative antiseptics. PMID- 16379448 TI - Using early troponin I testing to improve cardiac care. AB - Quicker test results following suspected myocardial infarction reduce the time required to make decisions on treatment, allowing it to begin earlier and facilitating a well-planned discharge. This article describes how the introduction of early diagnostic testing allowed the timely treatment of patients admitted to a medical admissions unit with chest pain. PMID- 16379449 TI - Oseltamivir. PMID- 16379450 TI - The role of the link teacher in the context of nurse education. AB - Link teachers have historically been employed to encourage the utilisation of nursing theory in the clinical setting. However, evidence suggests that this role has done little to help establish a theory-practice link. This article outlines the concept of a nurse academic linking with a clinical area, which is compatible with their clinical research activities. PMID- 16379451 TI - Rising to the challenge of autism. PMID- 16379452 TI - Work opportunities across the world. PMID- 16379453 TI - Sustaining promises for change. PMID- 16379454 TI - Hepatitis C and a midwife's experiences. PMID- 16379455 TI - Matters for discussion. PMID- 16379456 TI - Uniting midwives across Europe. PMID- 16379457 TI - One mother, one midwife. PMID- 16379458 TI - Confidentiality and teenage pregnancy--the affinity gap. PMID- 16379459 TI - Return to practice programme: progress update. PMID- 16379460 TI - Separated at birth--now joined at the hip! PMID- 16379461 TI - Evidence for episiotomy. PMID- 16379462 TI - Erinn Walton's letter in the September issue. PMID- 16379463 TI - Student documentary on the subject of in vitro fertilisation (IVF) and other forms of assisted conception. PMID- 16379464 TI - Anonymous letter from student in September's issue. PMID- 16379465 TI - Student midwife's letter in the September issue. PMID- 16379466 TI - "Are midwifery managers 'out of touch' with clinical practice"? PMID- 16379467 TI - Race rights. PMID- 16379468 TI - Last resort. PMID- 16379469 TI - Talking 2 ourselves. PMID- 16379472 TI - Should we interpret this as an intelligence deficit disorder that needs more education? PMID- 16379471 TI - Learning to live with voices. PMID- 16379473 TI - Many prisoners have asked if they can take a supply of tea bags with them when they leave. PMID- 16379475 TI - Recovery is about being involved with people--it's what you would want for yourself. Interview by Suzy Johnson. PMID- 16379476 TI - Don't shoot the messenger. PMID- 16379477 TI - Reaching out to refugees. PMID- 16379478 TI - Modernising day services. PMID- 16379479 TI - View point. Journalists get it wrong about mental health mostly through ignorance, not malice, argues Shelley Jofre. PMID- 16379481 TI - The death penalty and persons with mental disabilities: a lethal dose of stigma, sanism, fear of violence, and faulty predictions of dangerousness. PMID- 16379482 TI - Developmental venous anomalies and cavernous angiomas: a review of the concurrence, imaging, and treatment of these vascular malformations. AB - Deep venous anomalies and cavernous angiomas are two distinct intracranial lesions. Although these lesions can be found in association with each other, they have different pathologic characteristics and developmental mechanisms. These lesions can produce a variety of clinical symptoms or in some cases can be asymptomatic. The management of these anomalies can include conservative measures in some cases and aggressive intervention in others. Several imaging modalities can be used to evaluate these distinct lesions and often more than one are needed for a prompt and reliable diagnosis. Because of the tendency of these lesions to occur simultaneously, thorough evaluation of these anomalies and their surrounding tissues is critical in their clinical management. In this review, the characteristics and radiographic evaluation of these lesions will be presented and the therapeutic options for these anomalies will be discussed. PMID- 16379483 TI - The role of endoscopic ultrasound in the diagnosis and staging of pancreatic adenocarcinoma. AB - Pancreatic adenocarcinoma is the fifth leading cause of cancer-related death in the United States. Although spiral CT is the best noninvasive method for detecting pancreatic cancer, the sensitivity for small masses remains low. It is this group of patients with small masses who may most benefit from surgical resection. Endoscopic ultrasound employs the use of flexible endoscopic devices to perform diagnostic ultrasound imaging and is a more accurate means of detecting pancreatic tumors. The ability to perform cytological aspiration of pancreatic masses via EUS is also a major step in the diagnosis of pancreatic adenocarcinoma. This review article will summarize the role of endoscopic ultrasonography in the diagnosis and staging of pancreatic cancer as well as looking at its impact on the management of these malignancies. Additionally, we will discuss the utility of screening high-risk patients for pancreatic cancer using the tools that are currently available. PMID- 16379484 TI - Integrating caregiver health into patient care. AB - As the population of older adults continues to grow, there will be a simultaneous increase in the number of informal spousal and family caregivers. Given the demands of caregiving, informal caregivers are at risk for significant mental and physical health problems. Since many patients are dependent upon the caregiver to assist them with health-care needs, identifying caregiver needs and making appropriate referrals during patient visits is vital to ensuring quality care for your primary patient. The three main objectives for this review article are to 1) Provide an overview of negative mental and physical health consequences of caregiver stress, 2) Discuss the benefits to both patient and caregiver of addressing caregiver stress, and 3) Suggest ways in which caregiver stress can be identified and resources provided in health clinics with minimal staff time. A bibliography of resources for health care staff, patients, and families is included at the conclusion of the article. PMID- 16379485 TI - Place of service does make a difference. PMID- 16379486 TI - A matter of interpretation: what the ADA does and does not require for communicating with hearing-impaired patients. PMID- 16379487 TI - OSMA and Oklahoma medicine: 1966-1975. PMID- 16379488 TI - Patient Page #65. It's flu season! Protect yourself and others. PMID- 16379489 TI - Comparison of multiple auditory steady-state responses (80 versus 40 Hz) and slow cortical potentials for threshold estimation in hearing-impaired adults. AB - This study evaluated the use of multiple auditory steady-state responses (ASSRs) and slow cortical potentials (SCPs) to estimate behavioural audiograms in adults for compensation cases. Two groups of 23 subjects were assessed using either 80 Hz or 40 Hz multiple simultaneous tones with carrier frequencies of 0.5, 1.0, 2.0, and 4.0 kHz. SCP thresholds for 0.5, 1.0, and 2.0 kHz were obtained for both groups. Mean evoked potential thresholds (dB HL) minus behavioural pure-tone thresholds (dB HL) difference scores were 5-17 dB for the 80 Hz group, 1-14 dB for the 40 Hz group, and 20-22 dB for the SCPs. Thresholds for 40 Hz ASSR were significantly closer to behavioural thresholds than were 80 Hz or SCP thresholds. SCP and 40 Hz ASSR audiogram estimates were obtained more quickly than the 80 Hz ASSR. Multiple 40 Hz ASSR is the method of choice for evoked potential threshold estimation in adults. PMID- 16379490 TI - The post-auricular muscle response: an objective electrophysiological method for evaluating hearing sensitivity. AB - Post-auricular muscle responses (PAMRs) were recorded in sixteen adults with normal hearing and twenty adults with sensorineural hearing loss. Click stimuli were presented at 20 to 80 dB nHL via insert earphones. Only one ear was tested in hearing-impaired subjects, but normal-hearing subjects were tested monaurally and binaurally. PAMR amplitudes declined and latencies increased with decreasing click intensity. Both binaural stimulation and eye turn enhanced the PAMR. In hearing-impaired subjects, PAMR thresholds were correlated with audiometric thresholds for the eyes-turned condition. All normal-hearing subjects had PAMR when recording conditions were optimized and half had responses for the least optimal condition (20 dB nHL, monaural, eyes front). With eyes turned and monaural clicks at 35 dB nHL, the level widely used for infant hearing screening, most normal-hearing adults had a PAMR. Thus the PAMR is a robust response that may be a useful adjunct to ABR for objective hearing assessment. PMID- 16379491 TI - Rising-frequency chirps and earphones with an extended high-frequency response enhance the post-auricular muscle response. AB - The purpose of this study was to determine whether rising-frequency chirps presented via earphones with an extended high-frequency response would optimize the post-auricular muscle response (PAMR). The PAMR was recorded in adults using three different stimuli (a click, a rising-frequency chirp, and a truncated speech stimulus, /t/). Conventional ER-3A insert earphones were compared to ER-2 insert earphones to determine whether the PAMR is enhanced by the ER-2's extended highfrequency response. There were significant stimulus and earphone effects on PAMR amplitudes. The PAMR was largest for the chirp stimulus and the ER-2 earphones. The poorest responses were obtained using the /t/ stimulus and conventional ER-3A earphones. The results support previous ABR studies that have demonstrated a significant advantage of chirps over clicks for evoked response audiometry, and indicate that the PAMR is enhanced by inclusion of additional high-frequency stimulus energy. PMID- 16379492 TI - A comparison of the mismatch negativity and a differential waveform response. AB - A mismatch negativity response (MMN) and a new differential waveform were derived in an effort to evaluate a neural refractory or recovery effect in adult listeners. The MMN was elicited using oddball test runs in which the standard and deviant stimuli differed in frequency To derive the differential waveform, the same standard and deviant stimuli were presented alone. MMN responses were obtained by subtracting the averaged responses to standards from the deviants. The differential waveforms were obtained by subtracting the averaged responses to standards presented alone from deviants presented alone. Scalp topography for the MMN and differential waveforms were similar. A significant (p <.05) positive and negative correlation was found between the earlier and later components of the bimodal MMN and the NI and P2 component of the differential waveform, respectively. Further, N1 and P2 of the differential waveform were significant (p <.05) predictor variables of early and late peak amplitudes of the MMN. These results suggest that refractory effects may overlay/modify the morphology of the MMN waveform. PMID- 16379493 TI - Evaluation of phoneme compression schemes designed to compensate for temporal and spectral masking in background noise. AB - The effect of phonemic compression has been studied on speech intelligibility in background noise in hearing-impaired listeners with moderate-to-severe high frequency losses. One configuration, anti-upward-spread-of-masking (anti-USOM) focuses on a release from spectral masking of high-frequency speech cues by selective spectral tilting. Release from temporal masking is the main goal of a second configuration, high-pass filtered compression (HFC), which reduces the speech modulations within a high-pass filtered compression channel. Speech intelligibility was measured with consonant-vowel-consonant (CVC) words in a multi-talker babble and a single-talker background noise. Anti-USOM has a significant negative effect on the phoneme scores in background noise. HFC compression tends to improve vowel intelligibility in a single-talker background noise, especially for the listeners with a relatively poor speech score. In a multi-talker babble noise the effects of HFC compression tend to be negative. It can be concluded that no significant release from spectral or temporal masking is obtained by the applied processing. PMID- 16379494 TI - The preferred response slopes and two-channel compression ratios in twenty listening conditions by hearing-impaired and normal-hearing listeners and their relationship to the acoustic input. AB - This paper presents the results from two experiments in which normal-hearing and hearing-impaired listeners used an adaptive procedure to select their preferred frequency response slope and two-channel compression ratios in twenty listening conditions. Whereas the preferred response slope mostly depended on the difference in SNR between frequency bands, the preferred output levels in two channels depended highly on the intensity level entering each band. In both cases, subjects preferred less gain in frequency bands where noise was more intrusive and they preferred less gain for listening comfort than for speech understanding. The preferred response slope also depended on the slope of the audiogram. Relative to the prescribed NAL-RP response, the preferred gain variations improved the broadband SNR and hence listening comfort, but not the estimated speech intelligibility index. Overall, the findings confirm the approach used in many commercial products of applying wide dynamic range compression in multiple bands with additional gain reductions in bands where the noise is estimated to be dominant. PMID- 16379495 TI - Treatment of tinnitus in the elderly: a controlled trial of cognitive behavior therapy. AB - The aim of the study was to investigate the effects of cognitive behavioral therapy (CBT) in elderly people with tinnitus (<65 years). Thirty-seven patients were called in for a structured interview. Following exclusion, twenty-three participated in the trial. All participants underwent medical ear, nose, and throat (ENT) examination, audiometry, and tinnitus matchings. A randomized controlled design with a waiting list control group was used. A CBT treatment package was delivered in six weekly two hour group sessions. Outcome was measured using validated self-report inventories and daily diary ratings of annoyance, loudness and sleep quality for one week pretreatment, post-treatment. A three month follow-up was included at which time all participants had received treatment, but in a shorter format for the control group. Results showed statistically significant reductions of tinnitus-related distress. Thus, CBT was better than no treatment, but the particular aspects of CBT that contributed to the effects can not be established. In conclusion, the findings give some support for the use of group CBT for elderly people with tinnitus. PMID- 16379496 TI - Stepwise translocation of Dpo4 polymerase during error-free bypass of an oxoG lesion. AB - 7,8-dihydro-8-oxoguanine (oxoG), the predominant lesion formed following oxidative damage of DNA by reactive oxygen species, is processed differently by replicative and bypass polymerases. Our kinetic primer extension studies demonstrate that the bypass polymerase Dpo4 preferentially inserts C opposite oxoG, and also preferentially extends from the oxoG*C base pair, thus achieving error-free bypass of this lesion. We have determined the crystal structures of preinsertion binary, insertion ternary, and postinsertion binary complexes of oxoG-modified template-primer DNA and Dpo4. These structures provide insights into the translocation mechanics of the bypass polymerase during a complete cycle of nucleotide incorporation. Specifically, during noncovalent dCTP insertion opposite oxoG (or G), the little-finger domain-DNA phosphate contacts translocate by one nucleotide step, while the thumb domain-DNA phosphate contacts remain fixed. By contrast, during the nucleotidyl transfer reaction that covalently incorporates C opposite oxoG, the thumb-domain-phosphate contacts are translocated by one nucleotide step, while the little-finger contacts with phosphate groups remain fixed. These stepwise conformational transitions accompanying nucleoside triphosphate binding and covalent nucleobase incorporation during a full replication cycle of Dpo4-catalyzed bypass of the oxoG lesion are distinct from the translocation events in replicative polymerases. PMID- 16379497 TI - Two-photon imaging of cortical surface microvessels reveals a robust redistribution in blood flow after vascular occlusion. AB - A highly interconnected network of arterioles overlies mammalian cortex to route blood to the cortical mantle. Here we test if this angioarchitecture can ensure that the supply of blood is redistributed after vascular occlusion. We use rodent parietal cortex as a model system and image the flow of red blood cells in individual microvessels. Changes in flow are quantified in response to photothrombotic occlusions to individual pial arterioles as well as to physical occlusions of the middle cerebral artery (MCA), the primary source of blood to this network. We observe that perfusion is rapidly reestablished at the first branch downstream from a photothrombotic occlusion through a reversal in flow in one vessel. More distal downstream arterioles also show reversals in flow. Further, occlusion of the MCA leads to reversals in flow through approximately half of the downstream but distant arterioles. Thus the cortical arteriolar network supports collateral flow that may mitigate the effects of vessel obstruction, as may occur secondary to neurovascular pathology. PMID- 16379498 TI - Promoter variation in the DC-SIGN-encoding gene CD209 is associated with tuberculosis. AB - BACKGROUND: Tuberculosis, which is caused by Mycobacterium tuberculosis, remains one of the leading causes of mortality worldwide. The C-type lectin DC-SIGN is known to be the major M. tuberculosis receptor on human dendritic cells. We reasoned that if DC-SIGN interacts with M. tuberculosis, as well as with other pathogens, variation in this gene might have a broad range of influence in the pathogenesis of a number of infectious diseases, including tuberculosis. METHODS AND FINDINGS: We tested whether polymorphisms in CD209, the gene encoding DC SIGN, are associated with susceptibility to tuberculosis through sequencing and genotyping analyses in a South African cohort. After exclusion of significant population stratification in our cohort, we observed an association between two CD209 promoter variants (-871G and -336A) and decreased risk of developing tuberculosis. By looking at the geographical distribution of these variants, we observed that their allelic combination is mainly confined to Eurasian populations. CONCLUSIONS: Our observations suggest that the two -871G and -336A variants confer protection against tuberculosis. In addition, the geographic distribution of these two alleles, together with their phylogenetic status, suggest that they may have increased in frequency in non-African populations as a result of host genetic adaptation to a longer history of exposure to tuberculosis. Further characterization of the biological consequences of DC-SIGN variation in tuberculosis will be crucial to better appreciate the role of this lectin in interactions between the host immune system and the tubercle bacillus as well as other pathogens. PMID- 16379499 TI - Time course and cellular localization of SARS-CoV nucleoprotein and RNA in lungs from fatal cases of SARS. AB - BACKGROUND: Cellular localization of severe acute respiratory syndrome coronavirus (SARS-CoV) in the lungs of patients with SARS is important in confirming the etiological association of the virus with disease as well as in understanding the pathogenesis of the disease. To our knowledge, there have been no comprehensive studies investigating viral infection at the cellular level in humans. METHODS AND FINDINGS: We collected the largest series of fatal cases of SARS with autopsy material to date by merging the pathological material from two regions involved in the 2003 worldwide SARS outbreak in Hong Kong, China, and Toronto, Canada. We developed a monoclonal antibody against the SARS-CoV nucleoprotein and used it together with in situ hybridization (ISH) to analyze the autopsy lung tissues of 32 patients with SARS from Hong Kong and Toronto. We compared the results of these assays with the pulmonary pathologies and the clinical course of illness for each patient. SARS-CoV nucleoprotein and RNA were detected by immunohistochemistry and ISH, respectively, primarily in alveolar pneumocytes and, less frequently, in macrophages. Such localization was detected in four of the seven patients who died within two weeks of illness onset, and in none of the 25 patients who died later than two weeks after symptom onset. CONCLUSIONS: The pulmonary alveolar epithelium is the chief target of SARS-CoV, with macrophages infected subsequently. Viral replication appears to be limited to the first two weeks after symptom onset, with little evidence of continued widespread replication after this period. If antiviral therapy is considered for future treatment, it should be focused on this two-week period of acute clinical disease. PMID- 16379502 TI - Olanzapine for the management of pain in an adolescent girl. PMID- 16379500 TI - Exclusion and inclusion of nonwhite ethnic minority groups in 72 North American and European cardiovascular cohort studies. AB - BACKGROUND: Cohort studies are recommended for understanding ethnic disparities in cardiovascular disease. Our objective was to review the process for identifying, including, and excluding ethnic minority populations in published cardiovascular cohort studies in Europe and North America. METHODS AND FINDINGS: We found the literature using Medline (1966-2005), Embase (1980-2001), Cinahl, Web of Science, and citations from references; consultations with colleagues; Internet searches; and RB's personal files. A total of 72 studies were included, 39 starting after 1975. Decision-making on inclusion and exclusion of racial/ethnic groups, the conceptual basis of race/ethnicity, and methods of classification of racial/ethnic groups were rarely explicit. Few publications provided details on the racial/ethnic composition of the study setting or sample, and 39 gave no description. Several studies were located in small towns or in occupational settings, where ethnic minority populations are underrepresented. Studies on general populations usually had too few participants for analysis by race/ethnicity. Eight studies were explicitly on Caucasians/whites, and two excluded ethnic minority groups from the whole or part of the study on the basis of language or birthplace criteria. Ten studies were designed to compare white and nonwhite populations, while five studies focused on one nonwhite racial/ethnic group; all 15 of these were performed in the US. CONCLUSIONS: There is a shortage of information from cardiovascular cohort studies on racial/ethnic minority populations, although this has recently changed in the US. There is, particularly in Europe, an inequity resulting from a lack of research data in nonwhite populations. Urgent action is now required in Europe to address this disparity. PMID- 16379503 TI - Neuroleptic malignant syndrome associated with risperidone in a male with early onset schizophrenia. PMID- 16379506 TI - "Daddy, what is erectile dysfunction?" Direct-to-consumer advertising will be changing. PMID- 16379507 TI - Acute and long-term safety and tolerability of risperidone in children with autism. AB - Treatment-emergent adverse events (AEs) were monitored during an 8-week, double blind, placebo-controlled trial of risperidone (0.5-3.5 mg/day) in 101 children and adolescents with a lifetime diagnosis of autistic disorder. In addition, 37 placebo nonresponders received open-label risperidone for another 8 weeks. Of all the risperidone responders (n=65), 63 entered an open extension of another 16 weeks (6 months total risperidone exposure), and 32 of them were rerandomized to either continued risperidone therapy (n=16) or gradual replacement with placebo (n=16) over 8 weeks. We collected the following measures of safety and tolerability: (1) laboratory blood assessments (CBC with differential, electrolytes, and liver function tests) and urinalyses, (2) vital signs, (3) Side Effects Review of AEs thought to be associated with risperidone, (4) sleep records, (5) Simpson Angus Neurological Rating Scale (SARS), (6) Abnormal Involuntary Movement Scale (AIMS), and (7) height and weight. No clinically significant changes were found on the lab tests. During the 8-week acute trial, the most common AEs on the Side Effects Review, scored as moderate or higher, were as follows (placebo and risperidone, respectively): Somnolence (12% and 37%), enuresis (29% and 33%), excessive appetite (10% and 33%), rhinitis (8% and 16%), difficulty waking (8% and 12%), and constipation (12% and 10%). "Difficulty falling asleep" and anxiety actually favored the risperidone condition at statistically significant levels. The same AEs tended to recur through 6 months of treatment, although often at reduced levels. Using Centers for Disease Control (CDC) standardized scores, both weight and body mass index (BMI) increased with risperidone during the acute trial (0.5 and 0.6 SDs, respectively, for risperidone; 0.0 and 0.1 SDs, respectively, for placebo) and into open-label extension (0.19 and 0.16 SDs, respectively), although the amount of gain decelerated with time. Extrapyramidal symptoms, as assessed by the SARS, were no more common for drug than placebo, although drooling was reported more often in the risperidone group. There were no differences between groups on the AIMS. Two subjects had seizures (one taking placebo), but these were considered unrelated to active drug. Most AEs were mild to moderate and failed to interfere with therapeutic changes; there were no unanticipated AEs. The side effects of most concern were somnolence and weight gain. PMID- 16379508 TI - Risperidone-induced prolactin elevation in a prospective study of children, adolescents, and adults with mental retardation and pervasive developmental disorders. AB - OBJECTIVE: Risperidone is widely prescribed for aggression and self-injury in children, adolescents, and adults with mental retardation (MR) and pervasive developmental disorders (PDD). Risperidone elevates prolactin more than other atypical antipsychotic medications. Females may show greater prolactin elevation than males. METHOD: In this relatively long-term study of risperidone efficacy and safety for aggression and self-injury in children, adolescents, and adults with MR and PDDs, serum prolactin was measured in a 21-subject subset during the course of a double-blind, placebo-controlled trial. Prolactin was measured in ng/mL at baseline, once during acute treatment, and once during maintenance. RESULTS: In children and adolescents (n=10), mean age of 12.5 years, prolactin increased from mean 13.2+/-8.6 at baseline to 31.0+/-11.6 acutely and remained elevated at 37.9+/-10.4 in maintenance. In adults, mean age of 35.3 years, prolactin increased more markedly from 11.6+/-7.4 baseline (n=11) to 93.3+/-54.2 acutely but decreased to 67.8+/-62.9 in maintenance (n=7). Prolactin remained significantly elevated above normal in all subjects for at least 26 weeks. Mean prolactin of adult females, while similar to that of adult males at baseline, was 2.2 times male levels acutely and 3.7 times greater in maintenance. CONCLUSION: In this small subset, mean prolactin elevation persisted for at least 26 weeks. In adults, females showed significantly greater elevations than males. PMID- 16379509 TI - Elevated prolactin levels in male youths treated with risperidone and quetiapine. AB - The aim of this study was to report on the serum prolactin levels in 70 male youths at a residential treatment center who were treated with either risperidone or quetiapine. This is a cross-sectional retrospective medical chart review of 50 males (mean age, 13.5+/-2.8 years) treated with risperidone (mean dose, 2.4+/-1.6 mg/day) and 20 males (mean age, 13.5+/-2.4 years) treated with quetiapine (mean dose, 317.5+/-238 mg/day). Serum prolactin levels were drawn according to a protocol, after at least 6 weeks of treatment. Prolactin was above the upper limit of normal for 68% of the patients on risperidone and 20% of the patients on quetiapine (chi2 analysis: R>Q; p<0.001). Both risperidone and quetiapine produced dose-related increases in serum prolactin levels (R, r=0.34, p=0.017; Q, r=0.45, p=0.05). No correlation was found between duration of treatment and prolactin levels. Given that hyperprolactinemia secondary to antipsychotic treatment may result in reproductive and growth irregularities, periodic long term monitoring during treatment with these two atypical antipsychotics (and perhaps others as well) may be warranted. PMID- 16379510 TI - Rates and predictors of adherence with atypical antipsychotic medication: a follow-up study of adolescent inpatients. AB - BACKGROUND: The use of atypical antipsychotics is increasing in adolescent populations, but little is known about adherence with these treatments. This study examined postdischarge adherence of adolescents treated as inpatients with either olanzapine or risperidone. METHODS: Eighty-six (86) adolescent inpatients (43 per treatment) were contacted an average of 10 months (range, 90 days to 18 months) after discharge from the hospital. A structured interview examined compliance, side effects, and subjective impressions regarding the efficacy of treatment. Clinical symptoms rated by clinicians, self-reports from patients, hospital chart diagnosis, treatment group (risperidone/olanzapine), and demographic factors were used to predict compliance, as were the occurrence of side effects and subjective impressions of the efficacy of treatment. RESULTS: Forty-five percent (45%) of the patients were adherent with their medications, while only 12% discontinued treatment on their own, and 43% stopped medication at the request of their parent or a physician. Medication status and symptom severity did not predict nonadherence, and the only diagnosis related to adherence was substance abuse. The only side effect that predicted nonadherence was rapid weight gain during the hospitalization; this was slightly more common in olanzapine-treated patients. Nonadherent patients' subjective impressions of efficacy were no different from the patients who continued their medications, but the patients who stopped medication on their own were more likely to also have failed to attend recommended psychotherapy as well. IMPLICATIONS: Failure to continue recommended treatment with atypical antipsychotics was most common in cases who did not comply with other aspects of the discharge plan. Efficacy of the medication did not predict nonadherence, as half of the noncompliant subjects stated that the medication had been helpful. Rapid weight gain also predicted nonadherence, but subjective impression of long-term weight gain was not different across adherence or treatment status. These data tentatively suggest that in adolescent patients, tendencies toward general uncooperativeness and substance abuse may be at least as important as the occurrence of common side effects in the determination of medication adherence. PMID- 16379511 TI - Patterns of adherence to treatment in adolescents with bipolar disorder. AB - Despite relatively high rates of reported nonadherence in adults with bipolar disorder, no research has documented patterns of adherence in adolescents receiving treatment for this illness. This investigation sought to describe adherence in adolescents diagnosed with bipolar disorder and examine the relations between adherence, age, and chronicity of illness. Participants were 12 19 years of age and were receiving outpatient treatment for bipolar I, bipolar II, or bipolar disorder not otherwise specified (NOS). Parents were asked to estimate adherence to all prescribed treatments. Participants in this study were 38 adolescents (18 male, 20 female; mean age, 15.80 years, SD, 1.85). Parents reported an average of only 2.29 (SD, 2.90) missed medication dosages in the 1 month period prior to assessment. Full treatment adherence to a medication regimen, however, is reported in only 13 of 37 patients (34.2%) taking medication. Age is not associated with medication adherence. Participants with optimal adherence (no missed medication doses) are more recently diagnosed (M, 1.06 years; SD, 0.87) than patients who miss one or more medication doses (M, 3.12 years; SD, 3.36; t35=2.24; p=0.032). Rates of adherence in this sample of adolescents with bipolar disorder were somewhat higher than reports in adults and broadly consistent with findings in children with other psychiatric symptoms. Nonetheless, findings suggest vigilant monitoring of medication administration prior to assessing regimen effectiveness. PMID- 16379512 TI - Frequency of manic symptoms and bipolar disorder in psychiatrically hospitalized adolescents using the K-SADS Mania Rating Scale. AB - OBJECTIVE: The aim of this study was to assess the frequency of manic symptoms and bipolar spectrum disorders in an adolescent inpatient psychiatric sample using the Kiddie Schedule for Affective Disorders and Schizophrenia (K-SADS) Mania Rating Scale (MRS), parent and adolescent measures. METHOD: A total of 391 consecutive admissions to a psychiatric inpatient unit were assessed using the K SADS MRS, the Childhood Inventory of Psychiatric Syndromes (CHIPS), and other clinically relevant measures. RESULTS: The frequency of Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) manic symptoms in this population was generally high. However, only 19.6% received a consensus diagnosis of juvenile bipolar disorder. The K-SADS MRS discriminated bipolar from non-bipolar patients when parents rated symptoms but not when rated by adolescents. CONCLUSIONS: This study of non-selected adolescents over a one-year period demonstrates that bipolar spectrum disorders in an inpatient population are common, and that the use of the KSADS MRS is effective in identifying this syndrome. PMID- 16379513 TI - Adjunctive topiramate in hospitalized children and adolescents with bipolar disorders. AB - OBJECTIVE: The aim of this study was to assess topiramate as adjunctive treatment in children and adolescents hospitalized with bipolar disorders. METHODS: Medical records of all children and adolescents with a Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV-TR) (APA, 2000) diagnosis of bipolar disorder, type I, hospitalized for an acute manic, mixed, or depressive episode, were reviewed. The primary outcome measure was the Clinical Global Impression Severity (CGI-S) score. RESULTS: Twenty-five (25) children and adolescents received topiramate, with a mean final dose of 126 mg/day (range, 25-350 mg). Overall CGI-S scores significantly improved from 5.3+/-1.0 to 3.5+/-0.7, and mania CGI-S scores decreased from 5.4+/-1.0 to 3.3+/-0.9. Sixteen (16) of 25 (64%) bipolar patients were classified as responders (defined by an endpoint overall CGI-I score of less than or equal to 2). No serious adverse events occurred during treatment. Of 25 patients evaluated, 1 (4%) experienced mild sedation while treated with topiramate. CONCLUSIONS: Preliminary results of this retrospective chart review suggest that adjunctive topiramate may be associated with improvements in children and adolescents hospitalized for an acute manic, mixed, or depressive episode. Randomized and controlled trials with adjunctive topiramate in this population are needed to further explore this observation. PMID- 16379514 TI - Ethnic differences in maintenance antipsychotic prescription among adolescents with bipolar disorder. AB - PURPOSE: The aim of this study was to compare the longitudinal use of psychotropic medications in African-American and Caucasian adolescents with a diagnosis of bipolar disorder following their first psychiatric hospitalization. METHODS: Age- and gender-matched African-American (n=16) and Caucasian (n=16) adolescents, between the ages of 12 and 18 years and meeting Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) criteria for bipolar I disorder, manic or mixed, were evaluated over a period of 1 year following their first psychiatric hospitalization for a mood episode. Medication prescription and adherence during follow-up were compared between ethnic groups. RESULTS: African-American adolescents received antipsychotic prescription for a significantly greater percentage of time during follow-up than Caucasian adolescents, after adjusting for rate of psychotic features (79% versus 40%; p=0.05). African-American and Caucasian patients received mood stabilizer/anticonvulsant prescription for similar percentages of time during follow-up. African-American and Caucasian adolescents exhibited similar rates of full adherence to antipsychotics, mood stabilizers/anticonvulsants, and all psychotropic medications. CONCLUSIONS: African-American adolescents were prescribed atypical antipsychotic medications for longer periods of time than Caucasian adolescents, but reasons for this finding were unclear. Additional research efforts in adolescent bipolar disorder are needed to explore factors associated with ethnic disparity in pharmacological treatment interventions. PMID- 16379516 TI - Performance of clinician versus self-report measures to identify obsessive compulsive disorder in children and adolescents. AB - OBJECTIVE: The aim of this study was to study agreement between clinician-rated measures and self-report measures previously used in epidemiologic studies to identify obsessive-compulsive disorder (OCD) in youth and to determine the adequacy of self-report measures as screening instruments. METHOD: Leyton Obsessional Inventory-Child Version (LOI-CV) survey form self-report and Children's Yale-Brown Obsessive Compulsive Scale (CY-BOCS) clinician-report measures were compared in a sample of 81 OCD-affected youths diagnosed using structured diagnostic interviews and the best-estimate method. Sensitivities and agreement between tests are determined for different cutoff scores. RESULTS: The LOI-CV survey form total (item+interference) score is correlated with KSADS-E OCD severity (r=0.37, p=0.001) but not clinician-rated GAF scores (r=-0.18, p=0.12). LOI-CV survey form sensitivities at cutoff scores of 15, 20, 25, and 30 are 55%, 36%, 28%, and 19%, respectively. CY-BOCS scores are correlated with subject-rated interview severity (p<0.001) and clinician-rated GAF scores (por=0.03), but correlate with the CY-BOCS compulsion subscore. CONCLUSIONS: The clinician-rated CY-BOCS measure using parent and child reporting performs superiorly to the subject-rated LOI-CV measure using child reporting to identify pediatric OCD in a clinically referred population. Because self-report measures have been used in epidemiological studies, youth OCD prevalence rates may be higher than previously reported. PMID- 16379515 TI - An open trial of adjunctive donepezil in attention-deficit/hyperactivity disorder. AB - OBJECTIVE: Despite available pharmacotherapeutics, a number of youths with attentiondeficit/ hyperactivity disorder (ADHD) continue to experience residual symptoms and prominent executive function (EF) deficits resulting in impairment in multiple domains. We sought to determine if donepezil, used adjunctively to stimulant medication, would improve residual symptoms of ADHD and EF deficits. METHODS: In a 12-week open trial, we treated 7 children and 6 adults who had ADHD and evidence of further EF deficits with adjunctive donepezil. All subjects were stabilized on stimulants, at which time donepezil was initiated at 2.5 mg daily and increased to a maximum of 10 mg over the 12-week trial. RESULTS: Of 13 subjects receiving medication, 7 completed the trial. There was no clinically or statistically significant improvement in the ADHD Rating Scale and the Executive Function Checklist, our primary outcome measures. A majority of individuals experienced nonserious adverse events. CONCLUSIONS: Results of this small open study suggest that donepezil augmentation of stimulants is not well tolerated and does not appear useful for the treatment of residual ADHD and/or EF deficits. PMID- 16379517 TI - Oxcarbazepine in the treatment of child psychiatric disorders: a retrospective chart review. AB - Though not considered a first-line treatment, oxcarbazepine has become an option in the expanding effort to ameliorate severely dysregulated mood and behavioral symptoms in youth. Like most pharmaceuticals in child and adolescent psychiatry, oxcarbazepine is not U.S. Food and Drug Administration (FDA)-approved for the treatment of psychiatric disorders. A search of the world literature found a single case report pertaining to this topic. This paper is a chart review of 14 children and adolescents treated with oxcarbazepine who presented with moderate to severe problems with anger and irritability associated with a range of Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) diagnoses. A majority (70%) of patients was treatment-resistant to prior psychopharmacologic efforts, and 70% were receiving combined treatment with other agents in addition to oxcarbazepine. Moderate clinical global improvement was reported in 50% of patients receiving oxcarbazepine; tolerability was good in 86%. PMID- 16379518 TI - Overdoses and ingestions of second-generation antipsychotics in children and adolescents. AB - OBJECTIVE: We reviewed the available published data on intentional or unintentional secondgeneration antipsychotic overdoses in children and adolescents. The prescribing of secondgeneration antipsychotics has continued to increase over the past decade for children, adolescents, and adults. The authors reviewed the existing literature to determine the circumstances, presenting problems, treatment, and outcomes of youths who were exposed to nontherapeutic doses of these medications. METHODS: A systematic English-language Medline search of all reports (1989-2005) and a review of the bibliographies of all articles obtained was done to identify papers reporting an overdose or ingestion of a second-generation antipsychotic. Data were reviewed on clozapine, risperidone, olanzapine, ziprasidone, quetiapine, and aripiprazole. The annual reports of the American Association of Poison Control Centers National Data Collection System were reviewed from 1990 to 2003, the most recent report currently available. All fatalities in children and youths under 18 years of age were included. RESULTS: The literature review identified 40 reports that included 63 patients, ranging in age from 1 day to 17 years of age. The clinical presentations included drowsiness, lethargy, agitation, irritability, combativeness, and tachycardia. There were 11 fatalities in the cases reviewed, 1 from clozapine overdose, 3 from risperidone overdose, 2 from olanzapine overdose, and 5 from quetiapine overdose. All other cases reported no significant sequelae and resolved without any reported clinical consequences. Duration of overdose symptoms ranged from 24 hours to 7 days. One case of clozapine intoxication showed resolution of symptoms in 6 hours and, in another case of olanzapine overdose, symptoms resolved in 13 days. The most frequently employed treatments included intubation, gastric lavage, activated charcoal, intravenous fluids, artificial respiration, and restraints or sedatives. CONCLUSIONS: There is a need for future case reports to include serum medication level, weight of patient, coingestants, the health of the patient at baseline, relevant laboratory and toxicology studies and a standardized scale to rate the level of consciousness, such as the Glasgow Coma Scale. The existing pharmacovigilance data reports indicate these medications are relatively safe when taken in overdose, particularly when coingestants are not involved. PMID- 16379519 TI - Olanzapine overdose in children and adolescents: two case reports and a review of the literature. AB - Although the atypical antipsychotic olanzapine is increasingly being used in child and adolescent psychiatry, reports of olanzapine overdose in this young population are scarce. We report on two cases of adolescents who attempted suicide with an overdose of olanzapine: (1) A 14-year-old female ingested 275 mg olanzapine, which produced the highest reported nonlethal serum level (1503 ng/mL) and caused somnolence, agitation (acutely), and extrapyramidal symptoms (EPS; after 54 hours) but no major clinical complications. The serum olanzapine level dropped to 129 ng/mL within 48 hours; and (2) a 17-year-old male ingested 400 mg olanzapine, the highest reported nonlethal dose of olanzapine in adolescents, which produced respiratory suppression requiring intubation and mechanical ventilation; he recovered after 3 days. Based on clinical monitoring and postmortem data, the 2 patients survived the ingestion of high doses of olanzapine. We also provide a review of the literature, encompassing all reported cases of olanzapine overdose in children and adolescents and discuss symptoms, diagnosis, and treatment options, based on pharmacokinetic and pharmacodynamic considerations. PMID- 16379520 TI - Atomoxetine for the treatment of attention-deficit/hyperactivity disorder in children and adolescents with bipolar disorders. AB - Children and adolescents with bipolar disorder (BD) have a high incidence of comorbid attention-deficit/hyperactivity disorder (ADHD). In this paper, we report a consecutive case series on 7 patients with pediatric BD and ADHD who were treated with atomoxetine-and all but one were also treated in conjunction with mood stabilizers. All patients were outpatients at the Stanford Pediatric Bipolar Disorders Clinic. Information on patients was collected in a retrospective chart review. All but 1 patient demonstrated significant improvement in symptoms of ADHD. No patients had episodes of hypomania or mania during the treatment period. Adverse effects of atomoxetine treatment included sedation, nausea, and decreased appetite. These cases suggest that atomoxetine may be a safe and effective treatment for ADHD in conjunction with mood stabilizers in children with BD. PMID- 16379521 TI - A16-year-old girl with excited catatonia treated with low-dose oral Lorazepam. AB - A 16-year-old girl, who carried a diagnosis of schizoaffective disorder, presented with symptoms consistent with excited catatonia/delirious mania. We believe this is the first report of excited catatonia in a child under the age of 18. Her catatonic symptoms responded to low-dose oral lorazepam (higher doses led to worsening of her clinical status). This case highlights the importance of considering catatonia in the differential diagnosis of agitated delirium, even in pediatric populations, and it underscores the need for more data about the pathogenesis and treatment of catatonia. PMID- 16379522 TI - Macrocytosis associated with divalproex treatment. AB - Divalproex (DVP) is increasingly prescribed to children and adolescents in psychiatric practices. Among the hematologic adverse effects of DVP, decreased platelet count is well described in the medical literature. However, to date, few studies describe the occurrence of macrocytosis as an adverse effect of divalproex. We report two cases of pediatric patients who developed macrocytosis and decreasing platelet counts secondary to DVP treatment. Because macrocytosis remained an asymptomatic nonprogressive condition for our patients, we support the recommendation for closer surveillance of the patients' complete blood counts for development of anemia in addition to thrombocytopenia. PMID- 16379524 TI - Key science questions from the second conference on early Mars: geologic, hydrologic, and climatic evolution and the implications for life. AB - In October 2004, more than 130 terrestrial and planetary scientists met in Jackson Hole, WY, to discuss early Mars. The first billion years of martian geologic history is of particular interest because it is a period during which the planet was most active, after which a less dynamic period ensued that extends to the present day. The early activity left a fascinating geological record, which we are only beginning to unravel through direct observation and modeling. In considering this time period, questions outnumber answers, and one of the purposes of the meeting was to gather some of the best experts in the field to consider the current state of knowledge, ascertain which questions remain to be addressed, and identify the most promising approaches to addressing those questions. The purpose of this report is to document that discussion. Throughout the planet's first billion years, planetary-scale processes-including differentiation, hydrodynamic escape, volcanism, large impacts, erosion, and sedimentation-rapidly modified the atmosphere and crust. How did these processes operate, and what were their rates and interdependencies? The early environment was also characterized by both abundant liquid water and plentiful sources of energy, two of the most important conditions considered necessary for the origin of life. Where and when did the most habitable environments occur? Did life actually occupy them, and if so, has life persisted on Mars to the present? Our understanding of early Mars is critical to understanding how the planet we see today came to be. PMID- 16379525 TI - Caenorhabditis elegans survives atmospheric breakup of STS-107, space shuttle Columbia. AB - The nematode Caenorhabditis elegans, a popular organism for biological studies, is being developed as a model system for space biology. The chemically defined liquid medium, C. elegans Maintenance Medium (CeMM), allows axenic cultivation and automation of experiments that are critical for spaceflight research. To validate CeMM for use during spaceflight, we grew animals using CeMM and standard laboratory conditions onboard STS-107, space shuttle Columbia. Tragically, the Columbia was destroyed while reentering the Earth's atmosphere. During the massive recovery effort, hardware that contained our experiment was found. Live animals were observed in four of the five recovered canisters, which had survived on both types of media. These data demonstrate that CeMM is capable of supporting C. elegans during spaceflight. They also demonstrate that animals can survive a relatively unprotected reentry into the Earth's atmosphere, which has implications with regard to the packaging of living material during space flight, planetary protection, and the interplanetary transfer of life. PMID- 16379526 TI - Biosignatures from Earth-like planets around M dwarfs. AB - Coupled one-dimensional photochemical-climate calculations have been performed for hypothetical Earth-like planets around M dwarfs. Visible/near-infrared and thermal-infrared synthetic spectra of these planets were generated to determine which biosignature gases might be observed by a future, space-based telescope. Our star sample included two observed active M dwarfs-AD Leo and GJ 643-and three quiescent model stars. The spectral distribution of these stars in the ultraviolet generates a different photochemistry on these planets. As a result, the biogenic gases CH4, N2O, and CH3Cl have substantially longer lifetimes and higher mixing ratios than on Earth, making them potentially observable by space based telescopes. On the active M-star planets, an ozone layer similar to Earth's was developed that resulted in a spectroscopic signature comparable to the terrestrial one. The simultaneous detection of O2 (or O3) and a reduced gas in a planet's atmosphere has been suggested as strong evidence for life. Planets circling M stars may be good locations to search for such evidence. PMID- 16379527 TI - Bacillus subtilis spores on artificial meteorites survive hypervelocity atmospheric entry: implications for Lithopanspermia. AB - An important but untested aspect of the lithopanspermia hypothesis is that microbes situated on or within meteorites could survive hypervelocity entry from space through Earth's atmosphere. The use of high-altitude sounding rockets to test this notion was explored. Granite samples permeated with spores of Bacillus subtilis strain WN511 were attached to the exterior telemetry module of a sounding rocket and launched from White Sands Missile Range, New Mexico into space, reaching maximum atmospheric entry velocity of 1.2 km/s. Maximum recorded temperature during the flight was measured at 145 degrees C. The surfaces of the post-flight granite samples were swabbed and tested for recovery and survival of WN511 spores, using genetic markers and the unique DNA fingerprint of WN511 as recovery criteria. Spore survivors were isolated at high frequency, ranging from 1.2% to 4.4% compared with ground controls, from all surfaces except the forward facing surface. Sporulation-defective mutants were noted among the spaceflight survivors at high frequency (4%). These experiments constitute the first report of spore survival to hypervelocity atmospheric transit, and indicate that sounding rocket flights can be used to model the high-speed atmospheric entry of bacteria-laden artificial meteorites. PMID- 16379528 TI - A search for chiral signatures on Mars. AB - It is thought that the chiral molecules of living material can induce circular polarization in light at levels much higher than expected from abiotic processes. We therefore obtained high quality imaging circular polarimetry of the martian surface during the favorable opposition of 2003 to seek evidence of anomalous optical activity. We used two narrow-band filters covering 43% of the martian surface, 15% of it in-depth. With polarization noise levels <0.1% (4.3 upper limits 0.2-0.3%) and spatial resolution 210 km, we did not find any regions of circular polarization. When data were averaged over the observed face of the planet, we did see a small non-zero circular polarization 0.02%, which may be due to effects associated with the opposition configuration though it is at the limit of the instrumental capability. Our observations covered only a small fraction of parameter space, so although we obtained a null result, we cannot exclude the presence of optical activity at other wavelengths, in other locations, or at higher spatial resolution. PMID- 16379529 TI - Abiotic synthesis of organic compounds from carbon disulfide under hydrothermal conditions. AB - Abiotic formation of organic compounds under hydrothermal conditions is of interest to bio, geo-, and cosmochemists. Oceanic sulfur-rich hydrothermal systems have been proposed as settings for the abiotic synthesis of organic compounds. Carbon disulfide is a common component of magmatic and hot spring gases, and is present in marine and terrestrial hydrothermal systems. Thus, its reactivity should be considered as another carbon source in addition to carbon dioxide in reductive aqueous thermosynthesis. We have examined the formation of organic compounds in aqueous solutions of carbon disulfide and oxalic acid at 175 degrees C for 5 and 72 h. The synthesis products from carbon disulfide in acidic aqueous solutions yielded a series of organic sulfur compounds. The major compounds after 5 h of reaction included dimethyl polysulfides (54.5%), methyl perthioacetate (27.6%), dimethyl trithiocarbonate (6.8%), trithianes (2.7%), hexathiepane (1.4%), trithiolanes (0.8%), and trithiacycloheptanes (0.3%). The main compounds after 72 h of reaction consisted of trithiacycloheptanes (39.4%), pentathiepane (11.6%), tetrathiocyclooctanes (11.5%), trithiolanes (10.6%), tetrathianes (4.4%), trithianes (1.2%), dimethyl trisulfide (1.1%), and numerous minor compounds. It is concluded that the abiotic formation of aliphatic straight chain and cyclic polysulfides is possible under hydrothermal conditions and warrants further studies. PMID- 16379530 TI - Adsorption water-related potential chemical and biological processes in the upper martian surface. AB - Mars Odyssey has given strong evidence for the existence of water in the upper martian surface at equatorial latitudes. The water content, which corresponds to the hydrogen in the soil, can regionally reach values up to about 15%. This water is mainly in the form of structurally and partially irreversibly bound "crystal" water, and of reversibly bound and partially unfrozen adsorption water. This adsorption water, which has "liquid-like" properties as a two dimensional fluid or film, can trigger-in the presence of ultraviolet light and in concentrations similar to what has been measured on Mars-photocatalytic processes that are important for martian surface chemistry. The consequences of the diurnally variable presence of adsorption water on the chemistry and hypothetical biological processes at and in the upper martian surface at equatorial and mid latitudes are discussed in terms of water-related environmental aspects for chemical and hypothetical life processes on Mars. PMID- 16379531 TI - Venus, Mars, and the ices on Mercury and the moon: astrobiological implications and proposed mission designs. AB - Venus and Mars likely had liquid water bodies on their surface early in the Solar System history. The surfaces of Venus and Mars are presently not a suitable habitat for life, but reservoirs of liquid water remain in the atmosphere of Venus and the subsurface of Mars, and with it also the possibility of microbial life. Microbial organisms may have adapted to live in these ecological niches by the evolutionary force of directional selection. Missions to our neighboring planets should therefore be planned to explore these potentially life-containing refuges and return samples for analysis. Sample return missions should also include ice samples from Mercury and the Moon, which may contain information about the biogenic material that catalyzed the early evolution of life on Earth (or elsewhere). To obtain such information, science-driven exploration is necessary through varying degrees of mission operation autonomy. A hierarchical mission design is envisioned that includes spaceborne (orbital), atmosphere (airborne), surface (mobile such as rover and stationary such as lander or sensor), and subsurface (e.g., ground-penetrating radar, drilling, etc.) agents working in concert to allow for sufficient mission safety and redundancy, to perform extensive and challenging reconnaissance, and to lead to a thorough search for evidence of life and habitability. PMID- 16379532 TI - A block-free hidden Markov model for genotypes and its application to disease association. AB - We present a new stochastic model for genotype generation. The model offers a compromise between rigid block structure and no structure altogether: It reflects a general blocky structure of haplotypes, but also allows for "exchange" of haplotypes at nonboundary SNP sites; it also accommodates rare haplotypes and mutations. We use a hidden Markov model and infer its parameters by an expectation-maximization algorithm. The algorithm was implemented in a software package called HINT (haplotype inference tool) and tested on 58 datasets of genotypes. To evaluate the utility of the model in association studies, we used biological human data to create a simple disease association search scenario. When comparing HINT to three other models, HINT predicted association most accurately. PMID- 16379533 TI - An approximation algorithm for haplotype inference by maximum parsimony. AB - This paper studies haplotype inference by maximum parsimony using population data. We define the optimal haplotype inference (OHI) problem as given a set of genotypes and a set of related haplotypes, find a minimum subset of haplotypes that can resolve all the genotypes. We prove that OHI is NP-hard and can be formulated as an integer quadratic programming (IQP) problem. To solve the IQP problem, we propose an iterative semidefinite programming-based approximation algorithm, (called SDPHapInfer). We show that this algorithm finds a solution within a factor of O(log n) of the optimal solution, where n is the number of genotypes. This algorithm has been implemented and tested on a variety of simulated and biological data. In comparison with three other methods, (1) HAPAR, which was implemented based on the branching and bound algorithm, (2) HAPLOTYPER, which was implemented based on the expectation-maximization algorithm, and (3) PHASE, which combined the Gibbs sampling algorithm with an approximate coalescent prior, the experimental results indicate that SDPHapInfer and HAPLOTYPER have similar error rates. In addition, the results generated by PHASE have lower error rates on some data but higher error rates on others. The error rates of HAPAR are higher than the others on biological data. In terms of efficiency, SDPHapInfer, HAPLOTYPER, and PHASE output a solution in a stable and consistent way, and they run much faster than HAPAR when the number of genotypes becomes large. PMID- 16379534 TI - HOPE: a homotopy optimization method for protein structure prediction. AB - We use a homotopy optimization method, HOPE, to minimize the potential energy associated with a protein model. The method uses the minimum energy conformation of one protein as a template to predict the lowest energy structure of a query sequence. This objective is achieved by following a path of conformations determined by a homotopy between the potential energy functions for the two proteins. Ensembles of solutions are produced by perturbing conformations along the path, increasing the likelihood of predicting correct structures. Successful results are presented for pairs of homologous proteins, where HOPE is compared to a variant of Newton's method and to simulated annealing. PMID- 16379535 TI - Gene proximity analysis across whole genomes via PQ trees. AB - Permutations on strings representing gene clusters on genomes have been studied earlier by Uno and Yagiura (2000), Heber and Stoye (2001), Bergeron et al. (2002), Eres et al. (2003), and Schmidt and Stoye (2004) and the idea of a maximal permutation pattern was introduced by Eres et al. (2003). In this paper, we present a new tool for representation and detection of gene clusters in multiple genomes, using PQ trees (Booth and Leuker, 1976): this describes the inner structure and the relations between clusters succinctly, aids in filtering meaningful from apparently meaningless clusters, and also gives a natural and meaningful way of visualizing complex clusters. We identify a minimal consensus PQ tree and prove that it is equivalent to a maximal pi pattern (Eres et al., 2003) and each subgraph of the PQ tree corresponds to a nonmaximal permutation pattern. We present a general scheme to handle multiplicity in permutations and also give a linear time algorithm to construct the minimal consensus PQ tree. Further, we demonstrate the results on whole genome datasets. In our analysis of the whole genomes of human and rat, we found about 1.5 million common gene clusters but only about 500 minimal consensus PQ trees, with E. Coli K-12 and B. Subtilis genomes, we found only about 450 minimal consensus PQ trees out of about 15,000 gene clusters, and when comparing eight different Chloroplast genomes, we found only 77 minimal consensus PQ trees out of about 6,700 gene clusters. Further, we show specific instances of functionally related genes in two of the cases. PMID- 16379536 TI - Predicting physiological concentrations of metabolites from their molecular structure. AB - Physiological concentrations of metabolites can partly be explained by their molecular structure. We hypothesize that substances containing certain chemical groups show increased or decreased concentration in cells. We consider here, as chemical groups, local atomic configurations, describing an atom, its bonds, and its direct neighbor atoms. To test our hypothesis, we fitted a linear statistical model that relates experimentally determined logarithmic concentrations to feature vectors containing count numbers of the chemical groups. In order to determine chemical groups that have a clear effect on the concentration, we use a regularized (lasso) regression. In a dataset on 41 substances of central metabolism in different organisms, we found that the physical concentrations are increased by the occurrence of amino and hydroxyl groups, while aldehydes, ketones, and phosphates show decreased concentrations. The model explains about 22% of the variance of the logarithmic mean concentrations. PMID- 16379537 TI - The victor/FRST function for model quality estimation. AB - Scoring functions are widely used in the final step of model selection in protein structure prediction. This is of interest both for comparative modeling targets, where it is important to select the best model among a set of many good, "correct" ones, as well as for other (fold recognition or novel fold) targets, where the set may contain many incorrect models. A novel combination of four knowledge-based potentials recognizing different features of native protein structures is introduced and tested. The pairwise, solvation, hydrogen bond, and torsion angle potentials contain largely orthogonal information. Of these, the torsion angle potential is found to show the strongest correlation with model quality. Combining these features with a linear weighting function, it was possible to construct a robust energy function capable of discriminating native like structures on several benchmarking sets. In a recent blind test (CAFASP-4 MQAP), the scoring function ranked consistently well and was able to reliably distinguish the correct template from an ensemble of high quality decoys in 52 of 70 cases (33 of 34 for comparative modeling). An executable version of the Victor/FRST function for Linux PCs is available for download from the URL http://protein.cribi.unipd.it/frst/. PMID- 16379538 TI - Structure-approximating inverse protein folding problem in the 2D HP model. AB - The inverse protein folding problem is that of designing an amino acid sequence which has a particular native protein fold. This problem arises in drug design where a particular structure is necessary to ensure proper protein-protein interactions. In this paper, we show that in the 2D HP model of Dill it is possible to solve this problem for a broad class of structures. These structures can be used to closely approximate any given structure. One of the most important properties of a good protein (in drug design) is its stability--the aptitude not to fold simultaneously into other structures. We show that for a number of basic structures, our sequences have a unique fold. PMID- 16379539 TI - Gene sequencing in neonates and infants with the long QT syndrome. AB - The objective was to analyze the clinical and molecular findings in a cohort of neonates and infants with the autosomal dominant long QT syndrome (LQTS). Those affected face a high risk of ventricular arrhythmia resulting in syncope, seizure or sudden death. Blood samples submitted for molecular diagnostic studies on 7 infants were subject to DNA extraction and mutation analysis of 18 selected exons in 5 LQTS genes (KCNQ1, HERG, SCN5A, KCNE1, and KCNE2). We detected 11 mutations in these 7 patients. Four patients had 2 mutations in 1 gene (compound heterozygotes) or 2 different genes (digenic inheritance), while 3 patients had 1 mutation each. Except for 1 mutation in KCNE1, all other mutations were detected alone or in combination within HERG and the SCN5A genes. Four of the mutations we found are novel. The lethal nature of the LQTS demands careful attention to the family history and prompt and precise diagnosis and treatment with serious consideration of endocardial pacemaker implantation. While much larger studies are needed, our data suggest that compound heterozygotes or those with 2 mutations in different genes are likely to have a more severe LQTS including early manifestations in neonates and infants. PMID- 16379540 TI - Indirect CFTR mutation identification by PCR/OLA anomalous electropherograms. AB - Mutations of CFTR gene are responsible for cystic fibrosis (CF) and other clinical conditions such as congenital absence of the vas deferens (CAVD), chronic pancreatitis (IP), and idiopathic disseminated bronchiectasis (DBE) classified as CFTR-related disorders. The PCR/OLA assay is designed to detect 31 known mutations including the 24 most common CF mutations worldwide, as identified by the CF Consortium. In order to define the CFTR genotype a series of 1812 individuals from central-southern Italy with and without CF manifestations were screened by using the PCR/OLA assay. Here we report the description of five cases of anomalous electropherograms obtained after PCR/OLA analysis, that led to the identification, in the homozygous state, of two point mutations (D110H and S589N) not included in the assay test panel, a large gene deletion (CFTRdel14b_17b), and an exonic polymorphism (c.4002A > G). Haplotype and real time PCR analysis were also performed in the subject carrying the large CFTR deletion. The study demonstrates that the PCR/OLA assay, besides being an efficient and user-friendly method to screen known mutations in the CFTR gene, may also function as a mutation/polymorphism-scanning assay, at least for certain nucleotide changes located in some critical regions of the gene. PMID- 16379541 TI - Development of an oligochip for genotyping human leukocyte antigen-B51 and its clinical application. AB - Human leukocyte antigen (HLA) has been associated with Behcet's disease (BD), among which HLA-B51 is the most strongly associated genetic marker. The sandwich hybridization technique was applied in the design of the specific oligonucleotide probes to ensure the specific and accurate results. The probe-spotted chip was hybridized with the polymerase chain reaction (PCR) amplicons including nine suballeles (B*5101-B*5109) of exons 2 and 3 of HLA-B51 gene to determine the HLA B51 genotypes. The results were subsequently confirmed by (PCR-SSP) and sequencing and were identical to those from polymerase chain reaction-sequencing specific primers (PCR-SSP) in 27 patients with BD and 30 healthy controls. This suggests that we successfully developed the oligochip for Behcet's-associated gene HLA-B51, which can effectively and accurately identify the HLA-B51 genotypes. PMID- 16379542 TI - Prevalence of the 35delG mutation in the GJB2 gene of patients with nonsyndromic hearing loss from Croatia. AB - The aim of this study was to investigate the allelic frequency of 35delG mutation in patients with recessive, nonsyndromic hearing loss (NSHL) compared to normal hearing individuals in the Croatian population. For this purpose, we analyzed 27 unrelated individuals with nonsyndromic hearing loss and 342 healthy individuals. The method we used is based on the principle of polymerase chain reaction (PCR) mediated, site-directed mutagenesis, followed by a BsiYI digestion. Among patients with NSHL, the 35delG mutation was found on 51.85% alleles. Carrier frequency among healthy control individuals was 1 in 68.4 (1.5%). The patients, found to be wild-type, either in heterozygous or homozygous form, were further tested by direct sequencing. Among them, two different mutations were observed, W24X and 313del14. Relatively high prevalence of 35delG mutation among patients with NSHL indicate that it is an important cause of NSHL in Croatia. Early diagnosis by identification of the 35delG mutation would greatly improve genetic counseling, as well as treatment and management of deafness in Croatia. PMID- 16379543 TI - Referral and experience with genetic testing among women with early onset breast cancer. AB - The purpose of this study was to determine whether physicians refer women with early onset breast cancer for genetic testing for BRCA1 and BRCA2, and how women respond to being offered testing and use the results. A web-based survey was distributed to 1221 women with early onset breast cancer. The survey included 158 questions divided into the following sections: demographics, family history of cancer, medical history, treatment history, and experience with genetic testing. Of 551 women diagnosed since 1993 who responded to the survey (45.1%), less than half (45%, n = 246) had ever discussed genetic testing with their physician and/or been referred to see a genetic counselor. Women with a family history of cancer (53%) and Ashkenazi Jewish women (81%) were more likely to have been referred. Of those who had discussed testing, 60% had undergone or were interested in testing. Overall 92 women were tested and 19 (20.6%) of these tested positive for a deleterious BRCA1 or BRCA2 mutation. Fourteen (74%) who tested positive subsequently underwent prophylactic surgery. Satisfaction with counseling and the decision to be tested was high. Among women who were not offered testing, the fact that the test had not been offered by their physician (89%), and fear of discrimination (83%) were the two most frequently cited factors for lack of interest in testing. A substantial number of women are not being referred to genetic counseling and/or testing after a diagnosis of early onset breast cancer. Among those who were tested, there was high interest in prophylactic surgery after confirmation of a BRCA1/2 mutation. PMID- 16379544 TI - Recruitment, genetic counseling, and BRCA testing for underserved women at a public hospital. AB - Genetic counseling and testing for heritable susceptibility to breast cancer caused by mutations in BRCA genes are largely unavailable to underserved women in the United States. Starting in 2002 the UCSF Cancer Risk Program offered this service free of charge to poor and medically indigent women at San Francisco General Hospital (SFGH). One recruitment strategy was a single-page questionnaire in four languages administered to women waiting for mammograms at SFGH. This report analyzes our first 3 years of experience with the recruitment questionnaire and compares the patient demographics and BRCA test results at SFGH with a more typical population undergoing genetic counseling and testing at UCSF's Mt. Zion Hospital (MZH). To our knowledge this is the first comprehensive clinical service for hereditary breast cancer in a U.S. public hospital. The ethnic mix of all 350 patients counseled was Caucasian 49% (approximately 20% of Caucasians reported Ashkenazi Jewish ancestry), Latina, 26%; African American, 13%; and Asian/other, 12%. Compared to the MZH population, SFGH patients were more ethnically diverse, less educated and more likely to be unemployed. Of 72 patients tested for BRCA mutations, 51 (71%) were negative, 5 were BRCA1 positive, and 12 were BRCA2 positive. Four (1 Caucasian, 1 Latina, 2 African American) had a total of 13 BRCA variants of unknown significance (VUS). The ratio of BRCA1/BRCA2 positive SFGH patients (5/12) was reversed compared to MZH (119/91). We evaluated 4573 recruitment questionnaires and 280 (6%) were judged to represent a high risk of heritable cancer. After additional screening and referral negotiation, 74 were scheduled for counseling. We judged the recruitment questionnaire to be a feasible, efficient, and reasonably cost-effective way to identify women at high risk of hereditary cancer in a traditionally underserved population. Underserved populations present special challenges for genetic counselors because of large, geographically dispersed families, cultural taboos about cancer diagnoses, and social marginalization. Despite these complexities, the clinical service at SFGH has been well accepted by patients and staff. Our successful venture can serve as a model for other public hospitals contemplating this clinical service. PMID- 16379545 TI - Impact of genetic counseling and DNA testing on individuals with colorectal cancer with a positive family history: a population-based study. AB - What is the impact of a genetic evaluation for colorectal cancer susceptibility? We previously reported a study of individuals diagnosed with colorectal cancer at 60 years of age or less in a five-county area of New York including Rochester. Subjects reporting at least one first- or second-degree relative with colorectal cancer were invited to receive genetic counseling and DNA testing. Of the 37 persons tested, we previously reported that 6 were found to have deleterious mutations in MSH2 or MLH1. A mutation has since been found in a seventh subject in another laboratory. To evaluate the impact of the testing experience, we followed-up on 36 of these 37 subjects at 3 and 12 months after communicating their DNA test result. We ascertained their knowledge of colon cancer, whether they told relatives their test result, their surveillance behavior, and their mental health. Three months after receiving their result, subjects knew more about colon cancer if they had more cancer of all types in their family (p = 0.02). At 12 months, they knew more if they had been found to have a mutation (p < 0.001), were younger when DNA tested (p < 0.01), or were younger when diagnosed with cancer (p < 0.03). All but 1 of those found to have an abnormality told relatives. Relatives of 3 subjects in whom a mutation had been found came to us to be tested themselves. At 12 months, surveillance for colon and endometrial cancer was more adherent if there were more total cancers in the family (p < 0.05) or if the testee were more worried about cancer (p < 0.05). Self-assessed mental health at 12 months was better for those who were married (p < 0.05). This study suggests that individuals undergoing a genetic evaluation for an inherited susceptibility to colorectal cancer pursue recommended surveillance and inform relatives of their result. PMID- 16379546 TI - The effect of preventability and severity levels of a genetic disorder on desire to communicate genetic testing information to family members. AB - This study examined the impact of severity and preventability of a genetic disorder on belief that genetic test results should be shared with family members, desire to be tested, and opinions about physician?s role in sharing information. Participants were 359 undergraduate students who received one of four descriptions of a fictitious genetic disorder called hemocythemia (i.e., factorial combination of high versus low disease preventability and high versus low disease severity). Logistic regression analyses indicated that disease severity and preventability did not influence agreement that genetic information should be shared with family members, interest in being tested for the disorder, or agreement that physicians should share genetic information without permission. Those who read about the disorder as preventable were more likely to agree that their family members should be tested for the disorder (odds ratio [OR] = 1.82, p < 0.05). Females, African Americans, and those of other races were more likely to agree that they would want their family members tested for hemocythemia than males or European Americans (p < 0.05). Describing the disorder as preventable minimized the effect of the severity manipulation on perception of disease severity (p < 0.05). Understanding a disorder to be preventable may increase encouragement of family members to be tested and affect perceived seriousness of the condition. PMID- 16379547 TI - Polymerase chain reaction-based analysis using deaminated DNA of dodecamer expansions in CSTB, associated with Unverricht-Lundborg myoclonus epilepsy. AB - Progressive myoclonus epilepsy of the Unverricht-Lundborg type is an autosomal recessive disorder that is characterized clinically by myoclonic seizures and ataxia. The majority of affected individuals carry repeat expansions of a dodecamer in the promoter region of the cystatin B gene. The unusually high GC content of this tract is refractory to conventional polymerase chain reaction (PCR), and, as a result, a circumventive procedure involving the deamination of DNA with sodium bisulfite has been proposed. This study evaluates the effectiveness of this deamination modification for the detection of dodecamer repeat variants. An analysis of 258 healthy Japanese individuals revealed an allele with four copies of the dodecamer repeat with a frequency of 0.01, in addition to the more commonly observed two and three copy repeat alleles. Homozygous repeat expansions 600 and 680 base pairs in length were detected in the analyses of two affected individuals. For these cases, sequencing, along with an alternative PCR-stutter formation, revealed 41 and 48 copies, respectively, of the dodecamer repeat. The complete conversion of C to T was observed in the expanded tracts, indicating that no methylation occurred at the CpG sites. Based on these results, it was concluded that the use of deaminated DNA allows for a precise analysis of consecutive GC tracts. PMID- 16379548 TI - LARaLink 2.0: a comprehensive aid to basic and clinical cytogenetic research. AB - LARaLink 2.0 (Loci Analysis for Rearrangement Link) is an enabling web technology that permits the rapid retrieval of clinical cytogenetic and molecular data. New data mining capabilities have been incorporated into version 2.0, building upon LARaLink 1.0, to extend the utility of the system for applications in both the clinical and basic sciences. These include access to the Chromosomal Variation in Man database and the GEO database. Together these new resources enhance the user's ability to associate genotype with phenotype to identify potential gene candidates. Unlimited access for researchers exploring disease-gene relationships and for clinicians extending practice in patient care is available at LARaLink.bioinformatics.wayne.edu:8080/ unigene. PMID- 16379550 TI - Stability of lycopene during food processing and storage. AB - With an increasing understanding of the health benefit of lycopene, how to preserve lycopene during food processing and storage has caused much attention. Lycopene belongs to the carotenoid family and mostly exists in nature as the all trans form. Heat, light, oxygen, and different food matrices are factors that have an effect on lycopene isomerization and autooxidation. Lycopene may isomerize to mono- or poly-cis forms with the presence of heat or oil or during dehydration. Reisomerization takes place during storage. After oxidation, the lycopene molecule split, which causes loss of color and off-flavor. The effects of heat, oxygen, light, and the presence of oil on the stability of lycopene are uniform in much of the literature; however, controversy still exists on some details, such as the conditions causing the occurrence of isomerization, the optimal moisture, and temperature for storage. PMID- 16379551 TI - Effect of casein hydrolysate, prepared with protease derived from Aspergillus oryzae, on subjects with high-normal blood pressure or mild hypertension. AB - Casein hydrolysate, prepared with Aspergillus oryzae protease, contains angiotensin I-converting enzyme inhibitory peptides, such as Val-Pro-Pro and Ile Pro-Pro. We conducted a randomized, double-blind, placebo-controlled study to evaluate the effect of casein hydrolysate on the blood pressure of 144 subjects with high-normal blood pressure (n = 104) and mild hypertension (n = 40). Subjects were randomly assigned to two groups for a 12-week intake period. In the test group, both systolic (SBP) and diastolic (DBP) blood pressure decreased significantly compared with the placebo group: SBP/DBP significantly decreased from 138.2 +/- 6.5/84.4 +/- 5.3 mm Hg at week 0 to 132.3 +/- 7.3 (P < .001)/81.2 +/- 4.8 mm Hg (P < .001) at week 12. In the stratified analysis, the test product showed an antihypertensive effect in both the subject group with high-normal blood pressure and that with mild hypertension. No side effect was observed in any subjects in this study. These results demonstrate that the casein hydrolysate, prepared with A. oryzae protease, produced a significant reduction in blood pressure in a population of subjects with high-normal blood pressure or mild hypertension without an adverse event. PMID- 16379552 TI - Genistein inhibits insulin-like growth factor-I receptor signaling in HT-29 human colon cancer cells: a possible mechanism of the growth inhibitory effect of Genistein. AB - Genistein, a soy isoflavone, has attracted much attention for its chemopreventive properties. Overexpression and constitutive activation of receptor tyrosine kinases are frequent events in human cancer. Because genistein has previously been reported to decrease HT-29 cell growth, the present study compared the effects of genistein with daidzein on the protein levels of the members of the ErbB receptor family and insulin-like growth factor-I (IGF-I) receptor (IGF-IR). HT-29 cells were cultured in serum-free medium, with 0, 25, 50, or 100 micromol/L genistein, daidzein, and/or 10 nmol/L IGF-I. DNA synthesis was estimated by 5 bromo-2'-deoxyuridine incorporation. Apoptotic cells were analyzed by annexin-V staining followed by flow cytometry. Genistein inhibited viable HT-29 cell numbers, in a dose-dependent manner, whereas daidzein had no effect on cell growth. The decrease in cell growth caused by genistein was due to decreased DNA synthesis and apoptosis induction. Immunoblot analysis showed that neither genistein nor daidzein decreased the protein levels of either of the epidermal growth factor receptors, ErbB2 or ErbB3. Genistein did, however, decrease the IGF IR protein levels, whereas daidzein had no effect. Genistein did not change the protein levels of insulin-receptor substrate-1 (IRS-1), the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI3K), or Akt. Immunoprecipitation/western blot analyses revealed that genistein decreased IGF-I stimulated phosphorylation of IGF-IR and IRS-1, recruitment of p85 to IGF-IR, and phosphorylation of Akt. These results suggest that inhibition of cell proliferation and induction of apoptosis by genistein are mediated, at least in part, by its ability to inhibit IGF-IR signaling and the PI3K/Akt pathway. PMID- 16379553 TI - Effects of soy isoflavone and/or estrogen treatments on bone metabolism in ovariectomized rats. AB - This study investigated whether soy isoflavone intake, with or without estrogen treatment, can reduce postmenopausal bone loss, and whether soy isoflavones can be an alternative for estrogen replacement therapy using a postmenopausal osteoporotic rat model in which ovariectomized female rats were fed a low calcium, high fat diet. Nine-week-old female Sprague-Dawley rats were ovariectomized and then fed low (0.1%) calcium diets with or without soy isoflavone supplementation (80 or 160 ppm) for 6 weeks. Some ovariectomized rats were fed the same diets but also injected with estrogen (10 microg/kg of body weight) subcutaneously. Serum calcium and phosphate levels were normal in all rats. Serum alkaline phosphatase activities were not affected by the treatments. Serum tartrate-resistant acid phosphatase activities and urinary hydroxyproline levels were not different between experimental groups. Bone mineral (calcium and phosphorus) contents were increased in the rats supplemented with 80 ppm soy isoflavone or the rats treated with only estrogen without soy isoflavone. Therefore, the effect of 80 ppm soy isoflavone supplementation was the same as estrogen injection, but there was no beneficial effect from combining soy isoflavones and estrogen injections. When 160 ppm soy isoflavone was used, the benefits were lessened or disappeared altogether. These results suggest that appropriate soy isoflavone supplementation prevents postmenopausal bone loss without estrogen injection and may have efficacy as an alternative to estrogen therapy. PMID- 16379554 TI - Immunomodulating activity of arabinogalactan and fucoidan in vitro. AB - Many polysaccharides obtained from natural sources are considered to be biological response modifiers and have been shown to enhance various immune responses. Here, we investigated the immunomodulating effects of arabinogalactan (AG) and fucoidan (FU) in vitro. Mouse spleen lymphocytes became cytotoxic to tumor cells after culture with AG and FU at concentrations of 10-100 microg/mL. Also, AG and FU were mitogenic in spleen lymphocytes and peripheral macrophages. Macrophages treated with AG and FU (10-100 microg/mL) exhibited induced tumoricidal activity and increased phagocytosis, lysosomal enzyme activity, and production of nitrite, H2O2, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-6. However, AG and FU had little effect on the level of IL-1beta. Thus, the tumoricidal effect of AG- and FU-activated macrophages appeared to be mainly mediated by production of free radicals (NO and H2O2) and cytokines (TNF-alpha and IL-6). These data suggest that AG and FU are activators of lymphocytes and macrophages. This property may contribute to their effectiveness in the immunoprevention of cancer. PMID- 16379555 TI - Antimicrobial activity of berberine alone and in combination with ampicillin or oxacillin against methicillin-resistant Staphylococcus aureus. AB - Methicillin-resistant Staphylococcus aureus (MRSA) bacteria have been responsible for substantial morbidity and mortality in hospitals because they usually have multidrug resistance. Some natural products are candidates as new antibiotic substances. In the present study, we investigated the antimicrobial activity of berberine, the main antibacterial substance of Coptidis rhizoma (Coptis chinensis Franch) and Phellodendri cortex (Phellodendron amurense Ruprecht), against clinical isolates of MRSA, and the effects of berberine on the adhesion to MRSA and intracellular invasion into human gingival fibroblasts (HGFs). Berberine showed antimicrobial activity against all tested strains of MRSA. Minimum inhibition concentrations (MICs) of berberine against MRSA ranged from 32 to 128 microg/mL. Ninety percent inhibition of MRSA was obtained with 64 microg/mL or less of berberine. In the checkerboard dilution test, berberine markedly lowered the MICs of ampicillin and oxacillin against MRSA. An additive effect was found between berberine and ampicillin, and a synergistic effect was found between berberine and oxacillin against MRSA. In the presence of 1-50 microg/mL berberine, MRSA adhesion and intracellular invasion were notably decreased compared with the vehicle-treated control group. These results suggest that berberine may have antimicrobial activity and the potential to restore the effectiveness of beta-lactam antibiotics against MRSA, and inhibit the MRSA adhesion and intracellular invasion in HGFs. PMID- 16379556 TI - Purification and characterization of an alkaline serine protease producing angiotensin I-converting enzyme inhibitory peptide from Bacillus sp. SS103. AB - An alkaline serine protease that hydrolyzes soybean protein into strong angiotensin I-converting enzyme inhibitory hydrolysates was isolated from alkalophilic Bacillus sp. SS103 and purified. The enzyme was purified by ammonium sulfate precipitation followed by gel filtration, cationic exchange column chromatography, and anionic exchange column chromatography. When run on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing gel, the purified enzyme gave a 36-kDa band and pI 5.5, respectively. The enzyme showed maximum activity at pH 11.0 and 50 degrees C. This enzyme activity was highly inhibited by aprotinin, suggesting it belongs to the serine protease class of enzymes. The K (m) and V (max) of the enzyme, when casein was used for the substrate, were 9.7 x 10-4 mM and 244 microg/minute, respectively. From the results of this study, it is concluded that the purified alkaline protease isolated from Bacillus sp. SS103 should be further studied for production of biofunctional hydrolysates. PMID- 16379557 TI - Anticancer activities of pomegranate extracts and genistein in human breast cancer cells. AB - Previous studies have demonstrated the anticarcinogenic activity of pomegranate extracts and genistein in a series of human cancer cells. In the present study, the potential anticancer effects of pomegranate extracts and genistein on inhibition of cell proliferation and induction of apoptosis in human breast cancer cells was investigated. Human breast cancer cells (MCF-7) were cultured as monolayers in complete RPMI 1640 medium. The cells were cultured for 48 hours to allow growth and achieve about 80% confluence in 48-well culture plates, and then exposed to the agents for 24 hours in single and combination treatments. Post treatment growth rate and apoptosis induction were assessed by the use of a series of bioassays-lactate dehydrogenase and 3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (inner salt) for viability and cytotoxicity; acridine orange-ethidium bromide and terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling assays for induction of apoptosis. Both pomegranate extracts and genistein had significant (dose- and time-dependent) cytotoxic and growth inhibition effects on MCF-7 cancer cells. Both growth inhibition and cytotoxicity were significantly higher (P < .01) in the combination treatments than in the single treatments with either agent. The data revealed that both drugs in single and in combination treatments induced apoptosis in MCF-7 cells. Apoptotic induction in the combination treatments was significantly higher (P < .01) than in single treatments. Both pomegranate extracts and genistein inhibit the growth of MCF-7 breast cancer cells through induction of apoptosis, with combination treatment being more efficacious than single treatments. PMID- 16379558 TI - Physiological activities of garlic extracts as affected by habitat and solvents. AB - Physiological activities of Korean-grown garlic (GKG) and Chinese-grown garlic (GCG) were examined. Nitrite-scavenging activity (NSA), superoxide dismutase (SOD)-like activity, and electron-donating ability (EDA) of garlic extracted with water or with either 50% or 100% ethanol were measured. NSA was optimized at pH 1.2 and was highest in water and 50% ethanol extracts of both origins. SOD-like activities of water or 50% ethanol extracts from both Korea and China were 48.43 63.89% lower than those of 1% or 0.1% L-ascorbate solutions. SOD-like activities of GKG extracts were higher than those of GCG extracts, and those of water extracts of samples were highest. EDAs of GKG extracts were higher (32.51-43.74%) than those of GCG extracts, while those of both sample extracts were lower than 1% or 0.1% L-ascorbate solutions. PMID- 16379559 TI - Effect of D-003, a mixture of very-long-chain aliphatic acids purified from sugarcane wax, on cerebral ischemia in Mongolian gerbils. AB - D-003 is a mixture of very-high-molecular-weight aliphatic acids purified from sugar cane wax (Saccharum officinarum), which inhibits platelet aggregation and lipid peroxidation. The objective of the present study was to evaluate the effect of D-003 on cerebral ischemia induced by ischemia-reperfusion (I-R) in Mongolian gerbils. Two experimental series were conducted. The first series investigated the effects of D-003 on cerebral edema, neurological symptoms, and mortality in Mongolian gerbils with cerebral ischemia induced by I-R, while the second series investigated the effects on histological markers of cerebral injury, such as edema intensity (vacuolization) and cerebral necrosis. Animals were randomly distributed in five experimental groups: a sham-operated group experiencing surgical handling except the clamping and orally treated with Tween/water vehicle and four groups subjected to the I-R surgical procedure. One of these groups was treated with the same vehicle, and the other three groups received D-003 at 25, 100, and 200 mg/kg, respectively. All treatments were administered for 14 days. D 003 (200 mg/kg) significantly reduced the cerebral edema and clinical symptoms provoked by I-R compared with the positive control group, whereas lower doses (25 and 100 mg/kg) were not effective. Positive control animals showed an injury profile characterized by swelling (tissue vacuolization) and necrosis of neurons in all areas of the brain studied (frontal cortex, hippocampus, and striatum). The results of the histological study were consistent with those observed by determining cerebral edema and symptoms observation. Thus, D-003 at 200 mg/kg significantly reduced histological markers of brain injury (swelling and necrosis) compared with the control group. It is concluded that D-003 administered orally at 200 mg/kg for 14 days protected against cerebral damage caused by bilateral cerebral ischemia in Mongolian gerbils. PMID- 16379560 TI - An investigation of the analgesic and anti-inflammatory effects of Nigella sativa seed polyphenols. AB - Extracts obtained from the seeds of Nigella sativa are used as a spice or remedy for the treatment of various inflammatory diseases. The purpose of this study was to examine the analgesic and anti-inflammatory effects of its polyphenols. N. sativa seed polyphenols were prepared, and analgesic and anti-inflammatory effects were studied in mice and rats using the acetic acid-induced writhing, formalin, light tail flick, carrageenan-induced paw edema, and croton oil-induced ear edema tests. In the acetic acid-induced writhing test, oral administration of N. sativa polyphenols (NSP) decreased the number of abdominal constrictions. Both oral and intraperitoneal administration of NSP significantly suppressed in a dose dependent manner the nociceptive response in the early and late phases of the formalin test, and the effect on the late phase was more pronounced. Pretreatment with naloxone failed to reverse the analgesic activity of NSP in this test. NSP did not produce a significant analgesia in the light tail flick test in mice. Oral administration of NSP did not produce a significant reduction in carrageenan induced paw edema. However, when injected intraperitoneally, NSP inhibited paw edema in a dosedependent manner. NSP when applied topically failed to reduce croton oil-induced ear edema. These results suggest that NSP have analgesic and anti-inflammatory effects. The lack of analgesic effect of NSP in the light tail flick test and also the failure of naloxone to reverse the analgesia in the formalin test reveal that mechanisms other than stimulation of opioid receptors are involved. PMID- 16379561 TI - Combination chemoprevention of experimental gastric carcinogenesis by s allylcysteine and lycopene: modulatory effects on glutathione redox cycle antioxidants. AB - Combination chemoprevention by diet-derived agents is a promising strategy for protection against gastric cancer. We therefore evaluated the combined chemopreventive effect of S-allylcysteine (SAC), an organosulfur constituent of garlic, and lycopene, a major carotenoid present in tomatoes, against N-methyl-N' nitro-N-nitrosoguanidine (MNNG) and saturated sodium chloride (S-NaCl)-induced gastric carcinogenesis in Wistar rats. The animals were divided into eight groups of six animals each. Rats in group 1 were given MNNG by intragastric intubation on days 0 and 14 as well as S-NaCl every 3 days during weeks 0-3. Animals in groups 2-4, administered MNNG and S-NaCl as in group 1, received in addition SAC and lycopene alone and in combination, respectively, three times per week starting on the day following the first exposure to MNNG. Groups 5-7 were given the chemopreventive agents alone, whereas group 8 served as controls. The animals were sacrificed after an experimental period of 21 weeks. Measurement of lipid peroxidation and antioxidants of the glutathione redox cycle in the stomach, liver, and erythrocytes was used to monitor the chemopreventive potential of SAC and lycopene. In the tumor tissue, diminished lipid peroxidation was accompanied by an increase in reduced glutathione (GSH) and GSH-dependent enzymes, whereas in the liver and erythrocytes, enhanced lipid peroxidation was associated with antioxidant depletion. Although SAC and lycopene alone significantly suppressed the development of gastric cancer, administration of SAC and lycopene in combination was more effective in inhibiting MNNG-induced stomach tumors and modulating the redox status in the tumor and host tissues. The results of the present study validate the hypothesis that diet-derived chemopreventive agents such as SAC and lycopene in combination may interact synergistically with high efficacy and lessened toxicity against gastric cancer. PMID- 16379562 TI - Food seasoning spices mixture improves glucose metabolism and lipid profile in fructose-fed hyperinsulinemic rats. AB - Fructose feeding has been shown to induce insulin resistance in rats, associated with hyperinsulinemia, hyperglycemia, and hypertriglyceridemia. We have investigated the effect of administering food seasoning spices mixture (SM) on glucose, insulin, and lipids in circulation and carbohydrate enzymes in the erythrocytes of high fructose-fed rats. Additionally, we also measured the protein glycation status by assaying the levels of glycated hemoglobin, fructosamine, and plasma protein glycation. Male Wistar rats received a daily diet containing either 60% fructose or 60% starch (control). The rats were administered SM at three different doses (10, 30, or 50 mg/day per rat) orally 15 days later. At the end of the 45-day experimental period, fructose-fed rats showed significantly higher levels of plasma glucose and insulin, dyslipidemia, and alterations in enzyme activities. Treatment with SM significantly reduced plasma glucose and insulin levels and brought about a favorable lipid profile. In these rats, the activities of enzymes of glucose metabolism were normal. These effects were observed at all three doses of SM. High homeostasis model assessment (HOMA) values indicated insulin resistance in fructose-fed rats, while the HOMA values in SM-treated fructose-fed rats were comparable to those of control rats. We conclude that administration of SM improves glucose metabolism and plasma lipid profile in fructose-fed rats, possibly through improved insulin-sensitizing actions of the active constituents. PMID- 16379563 TI - Effects of flax fiber on laxation and glycemic response in healthy volunteers. AB - We investigated whether a flax supplement taken orally or baked in a bakery product would effect the physiological responses characteristic of soluble and insoluble fiber, i.e., laxation and glycemic response, respectively. In Study 1, 26 healthy young adults consumed up to 15 g of fiber from a proprietary flax fiber supplement or as a psyllium supplement for 2 weeks once usual fecal weights were established. Changes in dietary fiber intake and acceptability of both products were evaluated. An increase in fecal weight was found with both fiber treatments. Supplemental fiber at intakes of 9.0 g/day (flax) and 10.4 g/day (psyllium) gave fecal bulking capacity of about 2.9 and 4.8 g of fecal weight/g of fiber, respectively. In Study 2, the effect of flax bread versus control white bread on glycemic response was studied. Eleven fasting subjects completed four test periods (duplicate trials of each bread) under standardized glycemic testing conditions. Paired t tests were used to analyze test compared with control peak blood glucose values (6.6 +/- 0.9 mmol/L compared with 6.9 +/- 0.7 mmol/L, P < .05, respectively) and area under the curve (AUC) (669 +/- 53 compared with 693 +/- 57, P = .015, respectively). Peak blood glucose values and AUC were improved by ingestion of flax fiber in healthy subjects. In conclusion, a flax fiber supplement provides the benefits of soluble and insoluble fiber. PMID- 16379564 TI - Bound sugars in hepatic glycoproteins from male rats during dietary citrus bioflavonoid and/or ascorbic acid supplementation. AB - The purpose of these experiments was to determine whether quantitative changes take place in the attached sugars of hepatic (postmitochondrial) glycoproteins isolated from rats fed a diet supplemented with citrus bioflavonoids (an equal mixture of rutin, naringin, and hesperidin) (B) and/or ascorbic acid (C) for 40 days and 90 days. Statistically significant increases in body weights (P < .05) were observed in the B-only groups and liver weights in the CB group (P < .01 .05) after 40 and 90 days of feeding the experimental diets, while liver weights were decreased in the B-only groups after 40 days of feeding (P < .05). In the acid-soluble glycoprotein fraction, statistically significant decreases were seen in bound hexoses and fucose (P < .05) in the CB group after 40 days, and in bound fucose only after 90 days (P < .05). In the acid-insoluble glycoprotein fraction, statistically significant changes were seen in bound hexoses and fucose (P < .05) in the CB group after 40 days, and in bound fucose only in the CB group after 90 days (P < .05). There is an apparent overall decrease in the sugar-rich acid soluble glycoprotein fraction that is accentuated even further by combined CB supplementation. This decrease is more probable after 40 days than it is after 90 days of dietary supplementation. An adaptive phenomenon is suggested for maintaining the intracellular environment during periods of dietary citrus bioflavonoid and/or ascorbic acid supplementation. PMID- 16379565 TI - Elemental and nutritional analysis of sea buckthorn (Hippophae rhamnoides ssp. turkestanica) Berries of Pakistani origin. AB - Sea buckthorn (Hippophae rhamnoides ssp. turkestanica) is a very important medicinal plant in northern areas of Pakistan. The fruit of sea buckthorn is rich in nutrients and medicinal compounds such as vitamins, carotene, flavonoids, essential oil, carbohydrates, organic acids, amino acids, and minerals. In order to compare various populations of sea buckthorn for chemical composition, eight populations from different areas of northern Pakistan were compared using fruit characteristics. Phytochemical analysis of berries showed vitamin C (250-333 mg/100 g), seed oil (7.69-13.7%), oil in softer pulp (19.2-29.1%), phytosterol content of seed oil (3.3-5.5%), and anthocyanin (0.5-25 mg/L), while the mineral element composition analysis revealed high contents of potassium (140-360 ppm), sodium (20-80 ppm), calcium (70-98 ppm), magnesium (150-240 ppm), iron (40-150 ppm), and phosphorus (110-133 ppm). This study established sea buckthorn berries as a good source of biochemical and mineral elements. The high variation between different populations shows the potential of selecting and breeding of the raw material for various defined purposes. PMID- 16379566 TI - Comparative evaluation of hepatoprotective activity of carotenoids of microalgae. AB - The present study deals with evaluation of the hepatotoprotective activity of carotenoids from two well-known microalgae, Spirulina platensis and Dunaliella salina. Carotenoids were extracted in hexane:isopropyl alcohol (1:1 vol/vol) and fed orally in olive oil to Wistar albino rats at a dose of 100 microg/kg of body weight/day (in terms of carotenoids). The degree of hepatoprotection was measured by estimation of biochemical parameters like serum transaminases [serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT)], serum alkaline phosphatase, total albumin, and total protein. The results were compared with those for a control group, a CCl4-induced hepatic damage group, and a group treated with synthetic beta-carotene (all-trans) at the same dose. The protein content of the CCl4-treated group, which received normal diet and a dose of toxin, showed a significant decrease, i.e., 3.92 mg/mL, whereas the protein levels were higher, i.e., 6.96 and 6.32 mg/mL, in the case of the Dunaliella and Spirulina, respectively, carotenoid-treated groups. The CCl4 treated group shown higher activity of transaminases (128.68 units/mL SGPT and 171.52 units/mL SGOT). However, the activity of SGPT was 62.83 units/mL for Dunaliella and 76.83 units/mL for Spirulina, i.e., carotenoids of Dunaliella showed a higher degree of protection. For serum alkaline phosphatase, the standard beta-carotene value was 81.52 units/mL, compared with 84.46 units/mL for the CCl4-treated group; however, natural algal carotenoids yielded 38.45 units/mL (D. salina) and 44.73 units/mL (Spirulina). The total albumin value diminished with CCl4 treatment (2.46 mg/mL); the effect was highest for Dunaliella, followed by the Spirulina carotenoid-treated group. The results clearly indicate that carotenoids from Dunaliella possess better hepatoprotection compared with those from Spirulina. High-performance liquid chromatography of the carotenoids indicated that Spirulina contains only beta-carotene and Dunaliella contains other carotenoids and xanthophyll. The increase in protection with Dunaliella indicates that mixed carotenoids exhibit better biological activity than beta carotene alone. The results of this study indicate that carotenoids obtained from an algal source have a higher antihepatotoxic effect, compared with synthetic beta-carotene and with beta-carotene alone from a natural source. PMID- 16379567 TI - Participation of alpha2 receptors in the antinociceptive activity of quercetin. AB - The present work was planned to evaluate the antinociceptive activity of quercetin, its site of action, and the involvement of adrenergic receptors in mediating the antinociceptive activity in thermal and chemonociceptive assays. Eight groups were employed in this study: Groups I, II, and III received quercetin alone in different doses, group IV animals were administered clonidine, group V animals were co-administered quercetin and clonidine, group VI animals were administered yohimbine 30 minutes prior to administratin of quercetin, group VII animals were administered yohimbine 30 minutes prior to co-administration of quercetin and clonidine, and group VIII animals were administered 0.5% carboxymethylcellulose in saline (vehicle for quercetin). The thermal nociception was measured by tail flick and hot plate methods. Quercetin produced a significant analgesic effect in a dose-dependent manner. Co-administration of low doses of quercetin and clonidine produced a synergistic analgesic effect. Pretreatment with yohimbine significantly reversed quercetin-as well as clonidine/quercetin combination-induced effects. The results of this study reveal that quercetin induces an antinociceptive effect and that this effect involves primarily the modulation of adrenergic pathways. PMID- 16379568 TI - Antioxidant and antimycobacterial activities of Tabernaemontana catharinensis extracts obtained by supercritical CO2 + cosolvent. AB - In the present work the antioxidant and antimycobacterial activities were determined for extracts from Tabernaemontana catharinensis. The extracts' global yields were obtained using supercritical CO2 plus cosolvent. The cosolvents ethanol, isopropyl alcohol, methanol, and water and their mixtures were used. The extracts were fractionated and analyzed by thin-layer chromatography and gas chromatography/flame ionization detection. The antimycobacterial activity was measured against Mycobacterium tuberculosis, Mycobacterium avium, and Mycobacterium kansasii. The antioxidant activity was determined by the coupled reaction of beta-carotene and limonene acid. The average global yield was approximately constant (2.4 +/- 0.1%) for the alcoholic cosolvents and significantly larger (15 +/- 1%) for the cosolvent water and its alcoholic mixtures. The content of alkaloids in the extracts was strongly affected by the cosolvent. The antioxidant activity of the extracts ranged from 53% to 95%. The highest antimycobacterial activity was detected in the alkaloidal fraction (minimum inhibitory concentration = 128 microg/mL), while the lowest was verified in the aqueous fraction (minimum inhibitory concentration >512 microg/mL). PMID- 16379569 TI - Pro- and antioxidant effects and cytoprotective potentials of nine edible vegetables in southwest Nigeria. AB - Antioxidant and cytoprotective activities of boiled, cold, and methanolic extracts of nine edible vegetables in Southwest Nigeria were evaluated in the 1,1 diphenyl-2-picrylhydrazyl free radical assay and hemagglutination assay in bovine erythrocytes, respectively. Crassocephalum rubens showed the highest antioxidant activity (56.5%), Solanum americanum and Vernonia amygdalina exhibited moderate antioxidant activity (26.0-37.5% and 14.8-36.2%, respectively), Solanum macrocarpon, Telfaria occidentalis, Amaranthus hybridus, and Jatropha tanjorensis produced weak activity (1.6-15.8%, 1.6-7.7%, 2.8-6.62%, and 10.7-12.1%, respectively), while Celosia argentea and Talinum triangulare were pro-oxidants. It was also shown that extracts from all the vegetables are pro-oxidants at high concentrations of either 1 or 5 mg/mL or both. On the other hand, the studies on the cytoprotective effect showed that all the plant extracts demonstrated a very low hemagglutination titer value between 0.32 and 5.56 except S. americanum methanolic extract, which had a titer of 50.0. These results indicated correlation between the antioxidant properties and the hemagglutination values of these plant extracts; however, the membrane stabilizing capacity of the extracts supports the plants' antioxidant activity. PMID- 16379570 TI - Effect of supplementation of traditional medicinal plants on blood glucose in non insulin-dependent diabetics: a pilot study. AB - The effect of supplementation of a powdered mixture of three traditional medicinal plants-bittergourd, jamun seeds, and fenugreek seeds-in raw and cooked form on blood glucose was studied in 60 non-insulin-dependent male diabetics. The patients were divided into two groups of 30 each. The patients of group I were given the raw powdered mixture in the form of capsules; the patients of group II were given this mixture in the form of salty biscuits. Daily supplementation of 1 g of this powered mixture for a 1.5-month period and then a further increase to 2 g for another 1.5 months significantly reduced the fasting as well as the postprandial glucose level of the diabetic patients. A significant decrease in oral hypoglycemic drug intake and decline in percentage of the subjects who were on hypoglycemic drugs were found after the 3-month feeding trial. It was concluded that 2 g of a powdered mixture of traditional medicinal plants in either raw or cooked form can be successfully used for lowering blood glucose in diabetics. PMID- 16379571 TI - The effect of soybeans on the anovulatory cycle. AB - We treated 36 outpatients, suffering from secondary amenorrhea, who had no menstruation or no ovulation for more than 6 months before consulting our clinic. After polycystic ovary syndrome, hyperprolactinemia, and other ovarian disorders that require medical treatment had been ruled out through smear test examinations of the uterine cervix and uterine myoma, ovarian tumor, and endometriosis had been checked for with ultrasonography and serum CA-125, the subjects began to take 6 g/day of black soybean in micropowder form for 6 months (S group). We estimated the ovular improvement of theses patients, observing basal body temperature (BBT) and follicular development with ultrasonography during the menstrual cycle as the indexes for ovulation and compared them with 34 patients with no treatment (C group). In the S group, improved ovulation was seen in 12 patients, four patients became pregnant, and three patients had anovular menstruation within 3 months after starting to take soybean powder. The periods of first ovulation were 66 +/- 12 days. After ovulation started, all subjects had regular menstruations and ovulation, with more than a 7-day high phase in BBT. On the other hand, in the C group, improved ovulation was seen in two patients, and two patients had anovular menstruation. In conclusion, black soybean has the potential to improve the anovular menstrual cycle. PMID- 16379572 TI - Induction of extracellular matrix synthesis in normal human fibroblasts by anthraquinone isolated from Morinda citrifolia (Noni) fruit. AB - In previous studies we found that Morinda citrifolia (Noni) fruit extract up regulated biosynthesis of type I collagen and glycosaminoglycans in primary cultures of normal human fibroblasts. The objective of this study was to identify the active ingredients in Noni fruit extract. An active single compound having a type I collagen-stimulating effect was isolated and identified as 1,4-dihydroxy-2 methoxy-7-methylanthraquinone by nuclear magnetic resonance, infrared, and mass analysis. It was revealed that anthraquinone showed significantly increased elaboration of procollagen type I C-terminal peptide and glycosaminoglycans and reduced expression of the collagenase matrix metalloproteinase-1 dose-dependently in human dermal fibroblasts. Furthermore, in a clinical trial, a nano-emulsion containing anthraquinone predominantly increased the dermal type I procollagen in nude mouse skin. These results suggest that anthraquinone derived from Noni extract is a good candidate for use as a new anti-wrinkle agent due to its strong induction of biosynthetic activity of extracellular matrix components. PMID- 16379573 TI - Effects of soy protein and soy phytochemicals on mammary tumor development in female transgenic mice overexpressing human pituitary growth hormone. AB - The objective of this study was to determine the effect of soy-based diets on mammary tumors in female cancer- prone mice. Transgenic virgin female mice expressing human pituitary growth hormone and their respective phenotypically normal littermates were fed a diet containing either casein (C), low-isoflavone soy protein (LIS), or high-isoflavone soy protein (HIS). Indices of tumor development were measured throughout the study. Both days from birth until death and days on diet until death were increased [by 20% (P = .01) and 19% (P = .02), respectively] in the LIS group when compared with the C group. Both intervals were increased also (by 16% and 17%, respectively; P < .05) in the HIS group when compared with the C group. Days from birth to first tumor were increased by 7% (P < .05), as was days on diet to first tumor by 5% (P < .05), in the LIS group when compared with the C group. First-onset number of tumors was decreased (P = .02) by 41% and 34% in the LIS and C groups, respectively, when compared with the HIS group. Final onset of tumors was decreased (P < .05) by 44% and 9% in the LIS and HIS groups, respectively, when compared with the C group. Total area of final tumors was decreased (P < .05) by 30% in the LIS group when compared with the C group. Taken cumulatively, these data suggest that a diet rich in soy protein may provide protective benefits regarding tumor development in female cancer-prone mice. Furthermore, some bioactive compounds in the HIS diet appeared to confound the soy protein-induced beneficial effects. PMID- 16379574 TI - Hepatoprotective property of ethanolic and aqueous extracts of fluted pumpkin (Telfairia occidentalis) leaves against garlic-induced oxidative stress. AB - Fluted pumpkin (Telfairia occidentalis) leaf is a darkish-green leafy vegetable popularly used in soup and in herbal preparations for the management of many diseases in Nigeria. In this study, the hepatoprotective property of ethanolic and aqueous extracts of T. occidentalis leaf (earlier confirmed to have a high level of antioxidant activity) against garlic induced-oxidative stress in rat hepatocytes was investigated. Oxidative stress was induced in Wistar strain albino rats by overdosing them with raw garlic (4%) for 14 days, and this caused a significant increase (P < .05) in serum alkaline phosphatase (ALP), glutamate oxaloacetate transaminase (GOT), and glutamate pyruvate transaminase (GPT), while there was no significant change (P > .05) in serum bilirubin, albumin, globulin, and total proteins. However, intubation of some of the rats fed raw garlic with 5 mg or 10 mg/0.5 mL of T. occidentalis leaf extract (ethanolic or aqueous) caused a significant decrease (P < .05) in serum ALP, GOT, and GPT when compared with rats fed raw garlic without intubation with the T. occidentalis leaf extract. Moreover, 10 mg/0.5 mL of extract was more effective than 5 mg/0.5 mL of extract, while the aqueous extracts appeared to be more effective than the ethanolic extracts in protecting hepatocytes. It could be inferred that both aqueous and ethanolic extracts of T. occidentalis leaf have hepatoprotective properties, although the aqueous extract is more effective than the ethanolic extract, which could be attributed to the higher antioxidant activity of the aqueous extract than the ethanolic extracts of T. occidentalis leaves. PMID- 16379577 TI - Spinal cord diffusion tensor imaging and fiber tracking can identify white matter tract disruption and glial scar orientation following lateral funiculotomy. AB - Diffusion tensor magnetic resonance imaging (DTI) provides data concerning water diffusion in the spinal cord, from which white matter tracts may be inferred, and connectivity between spinal cord segments may be determined. We evaluated this potential application by imaging spinal cords from normal adult rats and rats that received cervical lateral funiculotomies, disrupting the rubrospinal tract (RST). Vitrogen and fibroblasts were transplanted into the surgical lesion at time of injury in order to fill the cavity. At 10 weeks, animals were sacrificed; the spinal cords were dissected out and then imaged in a 9.4-Tesla magnet. DTI tractography demonstrated the disruption of the rubrospinal tract axons while indicating which axon tracts were preserved. Additionally, DTI imaging could identify the orientation of glial processes in the gray matter adjacent to the site of injury. In the injured animals, reactive astrocytes in adjacent gray matter appeared to orient themselves perpendicular to white matter tracts. In summary, DTI identified not only white matter disruption following injury, but could distinguish the orientation of the accompanying glial scar. PMID- 16379576 TI - Methylprednisolone causes minimal improvement after spinal cord injury in rats, contrasting with benefits of an anti-integrin treatment. AB - Spinal cord injury (SCI) leads to complex secondary events that expand and exacerbate the injury. Methylprednisolone (MP) has been considered a standard of care for acute SCI. The purpose of this study was to test the effects of MP, in severe and more moderate severe clip-compression models of SCI, on the measures of neurological function and lesion sparing that we used previously to assess a highly effective anti-inflammatory therapy, a monoclonal antibody (mAb) to the CD11d integrin. Intravenous treatment with the anti-CD11d mAb blocks the infiltration of leukocytes into the lesion, limits secondary cord damage, and improves neurological outcomes. We also undertook a 2- week study of effects of these two therapies in combination. To permit direct comparison, the new findings with MP are presented together with reference to the previously published effects of the mAb. The severe SCI was at the 4(th) thoracic segment (T4), causing extensive motor dysfunction; the more moderate SCI was at T12 and caused less locomotor loss but the induction of mechanical allodynia. Neither MP alone nor the combination treatment improved Basso, Beattie, and Bresnahan 21-point open field locomotor scores at 2-12 weeks after SCI. These scores were ~4 points in the control, MP, and combination treatment groups, respectively, at 2 weeks after severe SCI at T4. By 6 weeks after T4 SCI, scores in the control and MP groups were ~7. At 12 weeks after the more moderate T12 injury, scores were ~8 in both control and MP treatment groups. MP treatment had no consistent effect on mechanical allodynia during 12 weeks after SCI. Control and MP-treated rats responded to approximately five of 10 stimuli to their backs and three of 10 stimuli to their hind paws. MP treatment increased areas of neurofilament and myelin near the injury site at T4 and T12. Thus, MP treatment spared tissue, but had no corresponding effect on neurological function. In contrast, the combination treatment did not spare myelin significantly. These neurological outcomes after treatment with MP contrast with the consistent and significant improvements after treatment with the anti-CD11d mAb. Effects of MP on the lesion were significant, but myelin sparing was less than that caused by the anti-CD11d mAb. The presence of MP in the combination therapy appeared to reverse the positive effects of the mAb. The poor neurological outcome after MP treatment may relate to the long-lasting reduction in hematogenous monocyte/macrophages within the injury site that it causes and to the prolongation of a neutrophil presence. These findings demonstrate that the non-selective and enduring effects of immunosuppressive therapy with MP not only fail to improve neurological outcomes, but also can block the beneficial actions of selective therapies such as the anti CD11d mAb. Combination treatments that cause intense immunosuppression should be viewed with caution. PMID- 16379578 TI - Increased close appositions between corticospinal tract axons and spinal sympathetic neurons after spinal cord injury in rats. AB - Treatments for spinal cord injury may promote new spinal cord synapses. However, the potential for new synapses between descending somatomotor and spinal sympathetic neurons has not been investigated. We studied rats with intact spinal cords and rats after a chronic, bilateral, dorsal spinal hemisection. We identified sympathetically related spinal neurons by transynaptic, retrograde transport of renally injected pseudorabies virus. We counted retrogradely labeled sympathetic preganglionic neurons (SPN) and putative sympathetic interneurons (IN) that, under light microscopy, appeared closely apposed by anterogradely labeled axons of the corticospinal tract (CST) and by axons descending from the well-established sympathetic regulatory region in the rostral ventrolateral medulla (RVLM). Spinal sympathetic neurons that were closely apposed by CST axons were significantly more numerous in lesioned rats than in unlesioned rats. CST axons closely apposed 5.4% of SPN and 10.3% of IN in rats with intact spinal cords, and 38.0% of SPN and 37.3% of IN in rats with chronically lesioned spinal cords. Further, CST appositions in SCI rats consisted of many more varicosities than those in uninjured rats. SPN and IN closely apposed by axons from the RVLM were not more numerous in lesioned rats. However, RVLM axons apposed many more SPN than IN in both control and lesioned rats. Therefore, RVLM sympathoexcitation may be mediated largely by direct synapses on SPN. Although we have not determined the functional significance of close appositions between the CST and spinal sympathetic neurons, we suggest that future studies of spinal cord repair and regeneration include an evaluation of potential, new, somatic-autonomic interactions. PMID- 16379579 TI - Changes in cerebral blood flow from the acute to the chronic phase of severe head injury. AB - We studied cerebral blood flow (CBF) in the transition from the acute to the chronic phase of severe head injury in order to determine patterns of change in relation to neurological outcome. We measured CBF with stable xenon-enhanced computed tomography (Xe-CT) in 20 consecutive patients at 1, 2, 3, 4, and 6 weeks after severe head injury, and analyzed the relation between the pattern of change in CBF and neurological outcome at 6 months after injury. CBF values were significantly lower in the brain-injured patients than in 14 healthy volunteers, except at 3 weeks after injury, when CBF increased in the patients to a value that did not differ significantly from that in the normal volunteers. We therefore focused on the change in CBF at 3 weeks after injury. We separated the 20 brain-injured patients into two subgroups, of which the first (subgroup A) consisted of nine patients whose CBF had returned to normal by week 3 post injury, while the second (subgroup B) consisted of 11 patients whose CBF was subnormal at week 3 post-injury. CBF was significantly higher in subgroup A than in subgroup B at 2 weeks post-injury (p < 0.05). CBF in subgroup B remained significantly lower than that in subgroup A throughout the study period. At 6 months post-injury, subgroup A had a significantly better neurological outcome than did subgroup B (p < 0.05). We conclude that patients whose CBF returns to normal at 2-3 weeks following severe traumatic brain injury after being abnormally low in the acute phase of injury can be expected to achieve a good neurological outcome. PMID- 16379580 TI - Clinical performance of NICE recommendations versus NCWFNS proposal in patients with mild head injury. AB - Recommendations on treatment of patients with head injury were recently proposed by the National Institute for Clinical Excellence (NICE). We tested the clinical performance of NICE variables versus the proposal of the Neurotraumatology Committee of the World Federation of Neurosurgical Societies (NCWFNS). Over a 5 year period, the clinical data of 7,955 adolescent and adult patients with mild head injury were prospectively collected and patients were managed according to the NCWFNS proposal. Outcome measures were (a) any post-traumatic lesion; (b) need for neurosurgical intervention; (c) unfavorable outcome (death, permanent vegetative state, severe disability) after 6 months. The predictive value of NICE variables was tested by logistic regression analysis. Three hundred fifty-four patients (6.8%) had intracranial lesions on computed tomography (CT) scan; neurosurgical intervention was needed in 108 patients (1.3%), and an unfavorable outcome occurred in 54 patients (0.7%) at 6-month follow-up. NICE variables were less sensitive than NCWFNS (93.5%; 95% confidence interval 91.0-95.2; vs. 97.8%; 96.1-98.7; p < 0.001), but far more specific (70.0%, 69.0-71.0, vs. 45.9%, 44.8 47.0; p < 0.001) for predicting intracranial lesions. NICE variables were also more specific (66.5%, 65.5-67.5, vs. 43.5%, 42.4-44.6; p < 0.001) in the prediction of neurosurgical intervention. 99.1% of unfavorable outcomes were predicted by both protocols. The CT order rate of NICE was much lower (34.1% vs. 57.1%; p < 0.001). In sum, the variables selected by NICE recommendations, when applied to a typical broad sample of emergency medicine, are a reliable, clinically sensible tool in predicting significant outcomes in patients with mild head injury and are resource saving. PMID- 16379581 TI - The effect of the selective NMDA receptor antagonist traxoprodil in the treatment of traumatic brain injury. AB - Traumatic brain injury (TBI) remains a major public health problem, and there is a great medical need for a pharmacological treatment that could improve long-term outcome. The excitatory neurotransmitter, glutamate, has been implicated in processes leading to neurodegeneration. Traxoprodil (CP-101,606) is a novel and potent glutamate receptor antagonist that is highly selective for the NR2B subunit of the NMDA receptor; it has been shown to be neuroprotective in animal models of brain injury and ischemia. A randomized, double-blind, placebo controlled study was therefore conducted to assess the efficacy and safety of a 72-h infusion of traxoprodil compared to placebo in subjects with computed tomography scan evidence of severe TBI (GCS 4-8). A total of 404 males and non pregnant females, aged 16-70, were treated within 8 h of injury. At baseline, subjects were stratified by motor score severity. The results showed that a greater proportion of the traxoprodil-treated subjects had a favorable outcome on the dichotomized Glasgow Outcome Scale (dGOS) at 6 months (delta 5.5%, OR 1.3, p = 0.21, 95% CI:[0.85, 2.06]) and at last visit (delta 7.5%, OR 1.47, p = 0.07, 95% CI:[0.97, 2.25]). The mortality rate with traxoprodil treatment was 7% less than with placebo treatment (OR 1.45, p = 0.08, 95% CI:[0.96, 2.18]). Differences between treatment groups were more pronounced in the severest subset (delta 11.8% for the dGOS at last visit and delta 16.6% for mortality). Traxoprodil was well tolerated. Although these results are intriguing, no definitive claim of efficacy can be made for traxoprodil for the treatment of severe TBI. PMID- 16379582 TI - A single dose, three-arm, placebo-controlled, phase I study of the bradykinin B2 receptor antagonist Anatibant (LF16-0687Ms) in patients with severe traumatic brain injury. AB - Traumatic brain injury (TBI) mortality and morbidity remains a public health challenge. Because experimental studies support an important role of bradykinin (BK) in the neurological deterioration that follows TBI, a double-blind, randomized, placebo-controlled study of Anatibant (LF16- 0687Ms), a selective and potent antagonist of the BK B(2) receptor, was conducted in severe (Glasgow Coma Scale [GCS] < 8) TBI patients (n = 25) at six sites in the United States. At 8-12 h after injury (9.9 +/- 2.8 h), patients received a single subcutaneous injection of Anatibant (3.75 mg or 22.5 mg, n = 10 each) or placebo (n = 5). The primary objective was to investigate the pharmacokinetics of Anatibant; general safety, local tolerability, levels of the bradykinin metabolite BK1-5 in plasma and cerebrospinal fluid (CSF), intracranial pressure (ICP), and cerebral perfusion pressure were also assessed. We observed a dose-proportionality of the pharmacokinetics, Cmax, and AUC of Anatibant. V(d)/F, Cl/F, and t(1/2) were independent on the dose and protein binding was >97.7%. Anatibant, administered as single subcutaneous injections of 3.75 g and 22.5 mg, was well tolerated in severe TBI patients with no unexpected clinical adverse events or biological abnormalities observed. Interestingly, plasma and CSF levels of BK1-5 were significantly and markedly increased after trauma (e.g., 34,700 +/- 35,300 fmol/mL in plasma vs. 34.9 +/- 5.6 fmol/mL previously reported for normal volunteers), supporting the use of Anatibant as a treatment of secondary brain damage. To address this issue, a dose-response trial that would investigate the effects of Anatibant on the incidence of raised ICP and on functional outcome in severe TBI patients is needed. PMID- 16379583 TI - Delayed transplantation of human neurons following brain injury in rats: a long term graft survival and behavior study. AB - The NTera2 (NT2) cell line is a homogeneous population of cells, which, when treated in vitro with retinoic acid, terminally differentiate into postmitotic neuronal NT2N cells. Although NT2N neurons transplanted in the acute (24 h postinjury) period survive for up to 1 month following experimental traumatic brain injury (TBI), nothing is known of their ability to survive for longer periods or of their effects when engrafted during the chronic postinjury period. Adult male Sprague-Dawley rats (n = 348; 360-400 g) were initially anesthetized and subjected to severe lateral fluid-percussion (FP) brain injury or sham injury. At 1 month postinjury, only brain-injured animals showing severe neurobehavioral deficits received cryopreserved NT2N neurons stereotaxically transplanted into three sites in the peri-injured cortex (n = 18). Separate groups of similarly brain-injured rats received human fibroblast cells (n = 13) or cell suspension vehicle (n = 14). Sham-injured animals (no brain injury) served as controls and received NT2N transplants (n = 24). All animals received daily immunosuppression for three months. Behavioral testing was performed at 1, 4, 8, and 12 weeks post-transplantation, after which animals were sacrificed for histological analysis. Nissl staining and anti-human neuronal specific enolase (NSE) immunostaining revealed that NT2N neurons transplanted in the chronic post injury period survived up to 12 weeks post-transplantation, extended processes into the host cortex and immunolabeled positively for synaptophysin. There were no statistical differences in cognitive or motor function among the transplanted brain-injured groups. Long-term graft survival suggests that NT2N neurons may be a viable source of neural cells for transplantation after TBI and also that these grafts can survive for a prolonged time and extend processes into the host cortex when transplanted in the chronic post-injury period following TBI. PMID- 16379584 TI - Neuroprotection following fluid percussion brain trauma: a pilot study using quercetin. AB - Previously, we were able to demonstrate the neuroprotective effect of quercetin in an animal model of acute traumatic spinal cord injury. The objective of the present study was to determine whether any neuroprotective effect is seen when quercetin is administered in an animal model of traumatic brain injury. Twenty six adult male Sprague-Dawley rats were submitted to moderate fluid percussion injury in the anterior midline position. Animals were divided into two experimental groups: one group received 25 mumol/kg quercetin starting 1 h after injury, while animals in the second group received saline vehicle (n = 13 per group). Eight animals were used as uninjured healthy controls. Eight animals in each experimental group were sacrificed at 24 h, while five animals per group were allowed to recover for 72 h following injury. Compound action potential amplitudes (CAPAs) were recorded on 400-microm vibrotome sections of the corpus callosum superfused with oxygenated artificial CSF (n = 3 per animal) in 20 experimental animals and five healthy controls. Three brains from animals in each experimental group and healthy controls were used for histological, immunocytochemical and biochemical analysis after sacrifice at 24 h. CAPAs in uninjured animals had a mean of 1.12 mV. This decreased to 0.55 mV in saline vehicle-treated injured animals by 24 h and changed little over the next 3 days. CAPAs were significantly better at 0.82 mV at 24 h and 0.76 mV at 3 days in quercetin-treated injured animals when compared to injured saline vehicle controls. Quercetin significantly prevented decrease of glutathione levels and decreased myeloperoxidase activity. We conclude that this dietary flavonoid has therapeutic potential following brain trauma. PMID- 16379585 TI - Electrical stimulation of the vagus nerve enhances cognitive and motor recovery following moderate fluid percussion injury in the rat. AB - Intermittent, chronically delivered electrical stimulation of the vagus nerve (VNS) is an FDA-approved procedure for the treatment of refractory complex/partial epilepsy in humans. Stimulation of the vagus has also been shown to enhance memory storage processes in laboratory rats and human subjects. Recent evidence suggests that some of these effects of VNS may be due to the activation of neurons in the nucleus locus coeruleus resulting in the release of norepinephrine (NE) throughout the neuraxis. Because antagonism of NE systems has been shown to delay recovery of function following brain damage, it is possible that enhanced release of NE in the CNS may facilitate recovery of function. To evaluate this hypothesis the lateral fluid percussion injury (LFP) model of traumatic brain injury was used and a variety of motor and cognitive behavioral tests were employed to assess recovery in pre-trained stimulated, control, and sham-injured laboratory rats. Two hours following moderate LFP, vagus nerve stimulation (30.0-sec trains of 0.5 mA, 20.0 Hz, biphasic pulses) was initiated. Stimulation continued in each animal's home cage at 30-min intervals for a period of 14 days, with the exception of brief periods when the animals were disconnected for behavioral assessments. Motor behaviors were evaluated every other day following LFP and tests included beam walk, locomotor placing, and skilled forelimb reaching. In each measure an enhanced rate of recovery and /or level of final performance was observed in the VNS-LFP animals compared to nonstimulated LFP controls. Behavior in the Morris water maze was assessed on days 11-14 following injury. Stimulated LFP animals showed significantly shorter latencies to find the hidden platform than did controls. Despite these behavioral effects, neurohistological examination did not reveal significant differences in lesion extent, density of fluorojade positive neurons, reactive astrocytes or numbers of spared neurons in the CA3 subarea of the hippocampus, at least at the one time point studied 15 days post-injury. These results support the idea that vagus nerve stimulation enhances the neural plasticity that underlies recovery of function following brain damage and provides indirect support for the hypothesis that enhanced NE release may mediate the effect. Importantly, since VNS facilitated both the rate of recovery and the extent of motor and cognitive recovery, these findings suggest that electrical stimulation of the vagus nerve may prove to be an effective non-pharmacological treatment for traumatic brain injury. PMID- 16379588 TI - Microarray analysis of VEGF-C responsive genes in human lymphatic endothelial cells. AB - Vascular endothelial growth factor-C (VEGF-C) is considered one of the most important factors influencing lymphatic endothelial cell biology. The goal of this work was to characterize the gene expression response by lymphatic endothelial cells (LECs) to VEGF-C. Primary cultures of human microvascular LECs were exposed to 100 ng/mL VEGF-C for 30 minutes and 6 hours, and their lysates were evaluated by microarray analysis to determine changes in mRNA expression induced by VEGF-C. Characteristic of a response to a growth factor stimulus, the largest number of differentially expressed genes were transcription factors and cell cycle related. A number of genes known to be important in angiogenesis, tumorigenesis and tumor invasion, and the transport of proteins, solutes, and lipids were also affected. Interestingly, a number of genes related to lipid metabolism as well as neurogenesis and neurodegeneration were also responsive to VEGF-C stimulation. Further analysis of these genes may not only provide insight into the molecular mechanisms underlying lymphangiogenesis and associated pathogenesis, but may also identify other important roles of VEGF-C. PMID- 16379589 TI - A comparison of four diagnostic criteria for lymphedema in a post-breast cancer population. AB - BACKGROUND: Breast cancer survivors are at life-time risk of developing lymphedema (LE). The goal of this research was to describe LE incidence over time among women treated for breast cancer. METHODS AND RESULTS: Limb volume changes (LVC) were evaluated by two measurement methods, circumferences and perometry, among 118 participants followed preoperative to 12 months postdiagnosis. Four diagnostic criteria were used: 200 mL perometry LVC; 10% perometry LVC; 2 cm circumferential increase; and report of heaviness or swelling, either "now" or "in the past year." Using 200 mL, the estimated LE rate was 24% (95% CI = 17% 32%) at 6 months, and 42% (31%-53%) at 1 year. Using 10% LVC, the estimated LE rate was 8% (2%-13%) at 6 months, and 21% (12%-30%) at 1 year. Using 2 cm, the estimated LE rate was 46% (36%-56%) at 6 months, and 70% (60%-79%) at 1 year. Based on reported symptoms of heaviness or swelling, the estimated LE rate was 19% (11%-26%) at 6 months, and 40% (30-59%) at 1 year. CONCLUSIONS: In the absence of a gold standard, we can only say that the different LE definitions are not equivalent, but cannot say which is "best". From this data, it appears that 10% LVC corresponds to a more conservative definition, whereas the 2 cm difference corresponds to a more liberal definition. These preliminary findings also document the importance of baseline (preoperative) anthropometric and symptom data and monitoring of changes over time. Further investigation of LE occurrence over an extended time period is warranted. PMID- 16379591 TI - Recent progress in our understanding of the pathogenesis of infantile hemangiomas. PMID- 16379592 TI - Update on the molecular genetics of vascular anomalies. AB - Genetic factors play a critical role in the pathogenesis of vascular anomalies. Significant advances have been made in recent years in identifying the genetic and molecular determinants of a variety of vascular anomalies using a molecular genetic approach. Several genes for vascular anomalies have been identified. These genes include AGGF1 for Klippel-Trenaunay syndrome, RASA1 for capillary malformations, KRIT1, MGC4607, PDCD10 for cerebral cavernous malformations, glomulin for glomuvenous malformations, TIE2 for multiple cutaneous and mucosal venous malformations, VEGFR-3, FOXC2, NEMO, SOX18 for lymphedema or related syndromes, ENG, ACVRLK1, MADH4 for HHT or related syndromes, NDP for Coats' disease, Notch3 for CADASIL, and PTEN for Proteus Syndrome. These findings have made genetic testing possible in some clinical cases, and may lead to the development of therapeutic strategies for vascular anomalies. Furthermore, these studies have identified critical genes involved in vascular morphogenesis, and provided fundamental understanding of the molecular mechanisms underlying vasculogenesis and angiogenesis. PMID- 16379593 TI - Circulating endothelial progenitor cells and vascular anomalies. AB - Recent findings regarding pathways of stem/progenitor cell involvement in adult blood vessel growth (postnatal vasculogenesis) suggest new theories for the pathogenesis of vascular anomalies. The somatic growth of vascular malformations and the mysterious pattern of proliferation and involution in infantile hemangioma can no longer be purely understood through the paradigm of angiogenesis. Molecular signals for postnatal vasculogenesis are being discovered in numerous animal models of cancer and ischemia, yet little research has addressed the importance of vasculogenesis in the growth of vascular anomalies. In this review, we discuss early studies that have investigated stem/progenitor cell involvement in the pathophysiology of infantile hemangioma and other congenital vascular anomalies. PMID- 16379595 TI - Lymphedema therapy in the vascular anomaly patient: therapeutics for the forgotten circulation. PMID- 16379594 TI - A role for Hox A5 in regulating angiogenesis and vascular patterning. AB - BACKGROUND: Homeobox (Hox) genes are transcriptional regulators which modulate embryonic morphogenesis and pathological tissue remodeling in adults via regulation of genes associated with cell-cell or cell extracellular matrix (ECM) interactions. We previously showed that while Hox 3 genes promote angiogenesis, Hox D10 inhibits this process. METHODS AND RESULTS: Here we show that another Hox family gene, Hox A5, also blocks angiogenesis but accomplishes this by targeting different downstream genes than Hox D10. Sustained expression of Hox A5 leads to down regulation of many pro-angiogenic genes including VEGFR2, ephrin A1, Hif1alpha and COX-2. In addition, Hox A5 also upregulates expression of anti angiogenic genes including Thrombospondin-2. Furthermore, we show that while Hox A5 mRNA is expressed in quiescent endothelial cells (EC), its expression is diminished or absent in active angiogenic EC found in association with breast tumors or in proliferating infantile hemangiomas. CONCLUSIONS: Together our results suggest that restoring Hox A5 expression may provide a novel means to limit breast tumor growth or expansion of hemangiomas. PMID- 16379596 TI - Quality of life in vascular anomalies. AB - Hemangiomas and vascular malformations that cause disfigurement or functional limitations may affect the quality life of the affected individual and their families. Negative reaction from strangers, social stigmatization, sadness, stress and low self-esteem are some of the commonly reported effects of having such lesions. Extensive and accurate education about the disease process and advice from support groups may improve quality of life. Psychological intervention may be indicated for some individuals and families. PMID- 16379597 TI - The role of pediatric cardiology in the management of hemangiomas. PMID- 16379598 TI - Literature watch. A genetic Xenopus laevis tadpole model to study lymphangiogenesis. PMID- 16379600 TI - Synchronous kinetics of CD4+ lymphocytes and viral load before the onset of oral candidosis and hairy leukoplakia in a cohort of Mexican HIV-infected patients. AB - An observational, prospective, longitudinal cohort study was performed at the AIDS Clinic of a tertiary care institution in Mexico City to determine the association of viral load (VL) and CD4+ lymphocyte kinetics with the development of oral candidosis (OC) and hairy leukoplakia (HL). Participants were HIV infected adult subjects, without a history of or current OC or HL, not receiving HAART. Oral examinations were performed at baseline and every month for evidence of OC or HL; CD4+ and VL determinations were done at baseline, at 6-month intervals, when oral lesions were detected, and 2 months later. Affected subjects (OL group) by OC or HL had clinical intervals defined before (antecedent), during (concurrent), and after their development. In the nonaffected individuals (NA group), 6-month intervals were determined. Differences (changes) along the clinical and study intervals were calculated for CD4+ and VL. The median study time was 178 (range: 31-924) days; 99 patients were included. The 2-year cumulative incidence of either oral lesion was 54% (49.5% for OC and 33.2% for HL). In the OL group (31 patients) a progressive and continuous decrease of CD4+ was found in the antecedent interval followed by a significant increase in VL in the concurrent period. The NA group showed a significant fall in CD4+ by semester 3, without a significant rise of VL in the following semester. The effect of CD4+ remained significant in a multivariate analysis. This study has shown that the onset of OC and/or HL is heralded by the sequence of a sustained reduction of CD4+, followed by a sharp increase of VL. In the multivariate analysis, the decrease in CD4+ lymphocytes appeared to be the predominant factor predicting the appearance of these oral lesions. Their potential use as markers of a recent change in the immunologic and virologic status of HIV-infected individuals is emphasized. PMID- 16379601 TI - Short communication: No detrimental immunological effects of mycophenolate mofetil and HAART in treatment-naive acute and chronic HIV-1-infected patients. AB - Mycophenolate mofetil has been proposed for HIV-1 therapy because of its guanine depleting effect, which is expected to interfere with HIV-1 replication directly by hampering reverse transcription and indirectly via inhibition of CD4+ T cell proliferation. However, treatment with mycophenolate mofetil might also compromise lymphocyte reconstitution and HIV-specific immunity. Therefore we longitudinally studied the effects of mycophenolate mofetil in combination with HAART on T cell proliferation, lymphocyte reconstitution, and HIV-specific CD4+ and CD8+ T cell responses in six therapy-naive, acute or chronic HIV-1-infected patients, as compared to eight patients treated with HAART alone. T cell proliferation in whole blood cultures of patients treated with mycophenolate mofetil was inhibited. Strikingly, ex vivo Ki67 expression within T cells was not influenced by treatment with mycophenolate mofetil. In vitro studies showed that Ki67 expression occurs at an early step of the cell cycle and was not inhibited by guanine depletion. When treatment with mycophenolate mofetil was stopped a transient increase in apoptosis and Ki67-expressing T cells was detected. This observation together with near complete inhibition of T cell proliferation in whole blood cultures during treatment with mycophenolate mofetil indicated that T cell proliferation was inhibited in patients treated with mycophenolate mofetil. Still, there was no evidence for detrimental effects of treatment with mycophenolate mofetil in addition to HAART on CD4+ T cell reconstitution or HIV specific immunity. PMID- 16379602 TI - Short communication: Influence of active tuberculosis on chemokine and chemokine receptor expression in HIV-infected persons. AB - Tuberculosis (TB) is the major opportunistic infection of HIV-1-infected patients in developing countries. Concurrent infection with TB results in immune cells having enhanced susceptibility to HIV-1 infection, which facilitates entry and replication of the virus. Cumulative data from earlier studies indicate that TB provides a milieu of continuous cellular activation and irregularities in cytokine and chemokine circuits that favor viral replication and disease progression. To better understand the interaction of the host with HIV-1 during active tuberculosis, we investigated in vivo expression of the HIV-1 coreceptors, CCR5 and CXCR4, and circulating levels of the inhibitory beta-chemokines, macrophage inflammatory protein-1-alpha (MIP-1alpha), macrophage inflammatory protein-1-beta (MIP-1beta), and regulated upon activation T cell expressed and secreted (RANTES), in HIV-positive individuals with and without active pulmonary tuberculosis. We found a significant decrease from normal in the fraction of CD4+ T cells expressing CCR5 and CXCR4 in individuals infected with HIV. However, CCR5 and CXCR4 expression did not differ significantly between HIV patients with and without tuberculosis. Higher amounts of MIP-1alpha, MIP-1beta, and RANTES were detected in plasma of HIV-1-positive individuals, particularly those with dual infection, although the increase was not found to be statistically significant. PMID- 16379603 TI - Coreceptor usage of primary HIV type 1 isolates obtained from different lymph node subsets. AB - Biological characteristics of virus quantitatively rescued from different cell types present in lymph nodes of HIV-1-infected individuals in various stages of their disease were determined, not including patients with AIDS defining illness. Viruses were obtained by cocultivation with donor monocyte-derived macrophages and T-lymphocytes and their biological phenotype compared to viruses obtained from the peripheral blood mononuclear cells of the same patient. The biological phenotype was determined on established cell lines (U937-2, CEM, and MT-2) and on the U87.CD4 coreceptor indicator cell lines and variable region 3 (V3) of the envelope was subjected to direct sequencing. All isolates obtained from lymph node subsets used CCR5 as coreceptor. Furthermore, these viruses were also sensitive to inhibition by beta-chemokines as analyzed for viruses of one patient. All 12 V3 regions showed a unique sequence indicating compartmentalization within each patient. The biological phenotype of CCR5 dependent (R5) HIV-1 isolates obtained from PBMC resembles the phenotype of viruses isolated from different lymph node cell subsets. PMID- 16379604 TI - Detection of HCV-RNA in saliva of HIV-HCV coinfected patients. AB - The presence of HCV-RNA in saliva of patients with chronic hepatitis C provides a biological basis for the potential transmission of this virus. HCV viremia is particularly high in HCV-HIV-coinfected patients, which could favor the presence of HCV in their saliva. This study was designed to evaluate the prevalence of HCV in saliva of HCV-HIV-coinfected patients. Stimulated whole saliva was collected from 75 HCV-HIV-coinfected patients and 75 HCV controls. The presence of HCV-RNA in saliva was tested by a highly sensitive noncommercialized nested PCR, and analyzed in relation to demographic, clinical, and analytical variables. HCVRNA was detected in the saliva of 49 (65%) HCV-HIV-coinfected patients and 39 (52%) HCV controls. The presence of HCV in saliva was not related to any of the analyzed variables in HCV-HIV-coinfected patients. In the HCV control group a statistically significant relationship was demonstrated only between the detection of HCV-RNA in saliva and the viral load in peripheral blood (p < 0.001). Our results indicate that there is a trend toward a higher HCV-RNA prevalence in the saliva of HCV-HIV-coinfected patients. PMID- 16379605 TI - Covariability of selected amino acid positions for HIV type 1 subtypes C and B. AB - We studied covariability of selected amino acid positions in globally dominant HIV-1 subtype C viruses. The analyzed sequences spanned the V3 loop, Gag p17, Gag p24, and five CTL epitope-rich regions in Gag, Nef, and Tat. The corresponding regions in HIV-1 subtype B were also evaluated. The analyses identified a great number of covarying pairs and triples of sites in the HIV-1B V3 loop (173 site pairs, 242 site triples). Several of these interactions were found in the earlier studies [e.g., the V3 loop covariability analyses by Korber et al. (Proc Natl Acad Sci USA 1993;90:7176-7180) and Bickel et al. (AIDS Res Hum Retroviruses 1996;12:1401-1411)] and have known biological significance. However, generally these key covarying sites did not covary in the HIV-1C V3 loop (total 17 covarying site pairs), suggesting that the V3 loop may have subtype differences in functional or structural operating characteristics. Covariability of positions 309 and 312 was observed in the immunodominant region HIV-1C Gag 291-320 but no covariability was found in the corresponding region of HIV-1B, and vice versa for Nef 122-141; these findings may reflect subtype-specific covariability within immunologically relevant regions. Gag p17 exhibited greater covariability and less diversity for HIV-1B than HIV-1C, raising the hypothesis that Gag p17 is highly immunodominant in HIV-1B and is especially important for HIV-1B vaccines. Information on covariability should be better exploited in assessments of HIV-1 diversity and how to surmount it with vaccine design. PMID- 16379606 TI - Characterization of nevirapine (NVP) resistance mutations and HIV type 1 subtype in women from Abidjan (Cote d'Ivoire) after NVP single-dose prophylaxis of HIV type 1 mother-to-child transmission. AB - Nevirapine (NVP) single dose is widely used in developing countries to prevent HIV-1 mother-to-child transmission. However, this regimen selects key drug resistance mutations that can impair further HAART efficacy. We studied the HIV-1 reverse transcriptase genotype from 29 Ivoirian women 1 month after an NVP single dose prophylaxis. NVP resistance mutations were observed in six (20.7%) women. The majority of the isolates were CRF02_AG. These results confirm previous studies and suggest the need for different prophylaxis regimens in this setting. PMID- 16379607 TI - Evaluations of HIV type 1 rev gene diversity and functional domains following perinatal transmission. AB - The human immunodeficiency virus type 1 (HIV-1) rev exons 1 and 2 sequences were analyzed from six mother-infant pairs following perinatal transmission. The rev open reading frame was maintained with a frequency of 93.96% in six mother-infant pairs' sequences. There was a low degree of viral heterogeneity and estimates of genetic diversity in mother-infant pairs' rev sequences. However, the distances of rev sequences between epidemiologically unlinked individuals were greater than in epidemiologically linked mother-infant pairs. Furthermore, phylogenetic parameters revealed that the epidemiologically linked mother-infant pairs were closer evolutionarily to each other as compared with epidemiologically unlinked mother-infant pairs. Both mothers and infants were under positive selection pressure as determined by the ratios of nonsynonymous to synonymous substitutions. The functional domains required for Rev activity, including nuclear export of RNA, RNA binding domain, and nuclear import signals, were conserved in all mother-infant pairs' sequences. The conservation of functional domains of rev and a low degree of heterogeneity following vertical transmission are consistent with an indispensable role of rev in the HIV-1 life cycle. PMID- 16379608 TI - The evolving molecular epidemiology of HIV type 1 among injecting drug users (IDUs) in Malaysia. AB - Earlier studies in the 1990s indicate that human immunodeficiency virus type 1 (HIV-1) subtype B has been the predominant subtype among injecting drug users (IDUs) in Malaysia. More recent studies performed between 2003 and 2004, however, show a high prevalence of unique CRF01_AE/B intersubtype recombinants among IDUs. To determine the subtype distribution among IDUs in Kuala Lumpur prior to the emergence of CRF01_AE/B intersubtype recombinants, the gag-pol or the reverse transcriptase gene was sequenced from IDUs who were diagnosed as HIV positive between 1993 and 2002. Subtype B was present at 50.0% followed by CRF01_AE/B recombinant at 41.7%, with more CRF01_AE/B recombinants detected between 2000 and 2002. All CRF01_AE/B recombinants shared similar recombination patterns. Interestingly, we found that this potential new candidate of circulating recombinant form (CRF) could have emerged as early as the mid-1990s. The results showed evidence of changing HIV-1 molecular epidemiology toward the predominance of CRF01_AE/B intersubtype recombinants among IDUs in Kuala Lumpur. PMID- 16379609 TI - Characterization of five nearly full-length genomes of early HIV type 1 strains in Ruili city: implications for the genesis of CRF07_BC and CRF08_BC circulating in China. AB - To trace the genesis of HIV-1 CRF07_BC and CRF08_BC, two predominant circulating recombinants among intravenous drug users in China, a retrospective molecular epidemiological investigation (1996-1998) was conducted in Ruili city of Yunnan, where the first AIDS epidemic among IDUs was reported in 1989. Fifty-four HIV-1 env C2V3 sequences were determined and genotyped with 49 subtype B and only 5 subtype C strains. The nearly full-length genome analyses of these five env-based subtype C samples revealed that four of them were actually BC recombinants and only one was pure subtype C. The first identified nonrecombinant HIV-1 subtype C, genetically close to Indian C representatives, provided direct evidence for the hypothesis that subtype C in China was introduced from India. Interestingly, three BC recombinants with subtype B as backbones were identified; one BC recombinant that precisely shared a common subtype B segment (nef region) with CRF07_BC and CRF08_BC was described, which indicated a close evolutionary relationship to these two CRFs. The sequences undoubtedly lead us to a better understanding of the emergence of CRF07_BC and CRF08_BC. PMID- 16379610 TI - A conservative domain shared by HIV gp120 and EIAV gp90: implications for HIV vaccine design. AB - Both HIV and EIAV belong to the retroviridae family and lentivirus genus. Two variable regions (V3 and V4) of equine infectious anemia virus (EIAV) gp90 and two variable regions (V1 and V2) of HIV gp120 possibly adopt the same topology. We have studied the N-glycosylation properties and B cell linear epitope distribution profile of these two regions. Our results indicated that V3 and V4 of EIAV gp90 are very similar to V1 and V2 of HIV gp120. The differences between EIAV virulent and vaccine strains are mainly located at these two regions. Vaccine strains lose two N-glycosylation sites at these two regions. Because of the conservative domain shared by EIAV gp90 and HIV gp120, V1 and V2 of HIV gp120 and their glycosylation properties should be the regions preferably considered for HIV vaccine design. PMID- 16379611 TI - No evidence for rapid subtype C spread within an epidemic in which multiple subtypes and intersubtype recombinants circulate. AB - There are multiple subtypes of HIV-1 circulating worldwide, but recently, subtype C has become highly prevalent, particularly in certain geographic regions. It is unclear whether the dominance of subtype C or other subtypes is due to increased fitness of certain subtypes for transmission, or a founder effect in new, rapidly growing epidemics. To examine whether the prevalence of one subtype increases over the course of an expanding epidemic that includes several circulating subtypes, we examined the distribution of HIV-1 subtypes in Kenya from 1986 to 2000. We found no evidence for an increase in the prevalence of subtype C, which remained low throughout this approximately 15-year period. Interestingly, the percentage of subtype D present in the population decreased significantly over that period, with a slight increase in subtype A. Throughout that period, intersubtype recombinant viruses were detected, including at the early stages of the epidemic. This latter finding suggests that reinfection may have occurred in high-risk groups early in the epidemic, leading to intersubtype recombinant viruses that underwent secondary spread. PMID- 16379612 TI - Analysis of HIV type 1 subtype C full-length gag gene sequences from India: novel observations and plausible implications. AB - Twenty-four HIV-1 gag genes from patients in India were sequenced and analyzed. All measured 1476-1491 nucleotides with an average of 1483 nucleotides in length. Phylogenetic analysis revealed a homogeneous epidemic of HIV-1 subtype C. Intragenotypic divergence of up to 6.6% was present. Fourteen novel conserved signature pattern residues were delineated for HIV-1 subtype C strains. Each of the 15 nucleocapsid (NC) basic residues was highly conserved p6 gag LXSLFG and PT/SAPP motifs were highly conserved except for PTVPT in three and PTAPT in two strains. Zinc finger motifs were conserved in all. Documented HIV-1 subtype C gag immunodominant CTL epitopes were conserved. The evolving predominance and the change in nature of the epidemic from Thai B to that of subtype C in the Northeastern regions of India were observed. Tracking the evolution of the Indian epidemic has implications for developing a vaccine. PMID- 16379613 TI - Effect of inherent variability of inhalation products on impactor mass balance limits. AB - When measuring the aerodynamic particle size distribution of pulmonary drug products, the commonly used instrument is a cascade impactor. For this type of analysis, a mass balance (MB) criterion, 85-115% of label claim, has been recommended by the U.S. Food and Drug Administration (FDA) to be included in the drug product specification. Using statistical model simulations, the effect of inherent product variability on the risk to fail the proposed criteria has been assessed and compared to the corresponding risk to fail the delivered dose uniformity (DDU) test. The results clearly show that the MB criterion is at odds with typical variability of orally inhaled products and seriously contributes to the risk that a typical batch would be rejected due to natural variability of the delivered dose of the product. The MB criterion is generally more difficult to comply with compared to the corresponding delivered dose uniformity (DDU) test, indicating that the proposed FDA MB specification overrules the DDU criteria as being that controlling the DDU. PMID- 16379614 TI - Influence of particle size and patient dosing technique on lung deposition of HFA beclomethasone from a metered dose inhaler. AB - The objective of this study was to determine the lung delivery of HFA-134a beclomethasone dipropionate (HFA-BDP) from a breath-activated inhaler (QVAR Autohaler) compared with proper and improper press and breathe (QVAR P&B) metered dose inhaler (MDI) technique. The hypothesis was that that the smaller particles of BDP from HFA-BDP would stay suspended longer in the inspiratory air of patients and thus reduce the deleterious effects of inhaler discoordination. The study was an open label, four period, cross-over design. Asthmatic patients (n = 7) with a history of asthma symptoms, an FEV-1 of >70% of predicted normal, and a history of reversibility to a beta-agonist of >or=12% were utilized. BDP was radiolabeled with technetium-99m and delivered from the QVAR Autohaler or QVAR P&B device in patients trained to reproducibly utilize coordinated and discoordinated P&B MDI technique. Patients using Autohaler MDI exhibited 60% lung deposition of BDP. Patients using coordinated technique with the P&B MDI exhibited 59% lung deposition. Patients trained to consistently actuate the P&B MDI before inhaling exhibited 37% lung deposition. Patients trained to consistently actuate the P&B MDI late in the inspiration (i.e., 1.5 sec into a 3 sec inspiration) exhibited 50% lung deposition. In conclusion, the breath activated Autohaler automatically provided optimal BDP lung deposition of 60%. Patients with good P&B MDI technique also received optimal lung deposition of 59%. The degree of lung deposition was decreased as patients demonstrated poor inhaler technique. However patients with poor technique still received a large lung dose of BDP (i.e., >or=37%) compared with lung deposition values of 4-7% for CFC-BDP MDIs previously published and confirmed in this study. PMID- 16379615 TI - Lung deposition after electronically breath-controlled inhalation and manually triggered conventional inhalation in cystic fibrosis patients. AB - The present work aimed to investigate whether lung deposition can be improved by using a device that optimizes the breathing pattern through electronic control. The relative lung deposition was estimated by inhalation of the marker substance, sodium cromoglycate (SCG), and measurement of urinary excretion of SCG. Thirteen cystic fibrosis (CF) patients (aged 8-20 years) received 20 mg of SCG as nebulizer solution by means of (a) conventional inhalation (Parimaster + Pari LC Star nebulizer, manual interrupter) and (b) electronically breath-controlled inhalation (AKITA + Pari LC Star nebulizer). Inhalations were trained and supervised by a physiotherapist. Patients were asked to provide answers to a questionnaire about the convenience of electronically breath-controlled inhalation. Urine was collected in five fractions until 12 h p.a., and the excreted SCG was determined by means of high-performance liquid chromatography (HPLC). Following breath-controlled inhalation, the amount of SCG excreted in urine was significantly greater than after conventional inhalation (2.22 +/- 0.61 mg vs. 1.63 +/- 0.59 mg, p < 0.001). The absorption half-life for SCG following breath-controlled inhalation was significantly shorter when compared with conventional inhalation (78 +/- 23 min vs. 107 +/- 29 min; p < 0.01), suggestive of a more peripheral deposition for the former. Ninety-two percent of the patients judged that the electronically breath-controlled inhalation was good or very good. In conclusion, inhalation with an electronically optimized breathing pattern yields a greater and more peripheral lung deposition of SCG compared with the manually triggered conventional nebulizer technique in CF patients with several years of aerosol inhalation experience. PMID- 16379616 TI - Relationship of stage mensuration data to the performance of new and used cascade impactors. AB - Cascade impaction is a standard test method for characterizing the quality of inhalable drug products. The sizes of the nozzles on each stage of the impactor are the critical dimensions for the performance of the impactor. Compendial reference methods call for periodic measurement of the size of the nozzles on each stage, a procedure known as stage mensuration. There is however currently no guidance on acceptable mensuration criteria. We aim to remedy this situation by providing a sound basis for understanding and using mensuration data, be it for acceptance criteria for new impactors or for the setting of mensuration tolerances for in-use impactors. We first show that multi-nozzle impactor stages behave as if all of the nozzles are equal in size to an effective diameter, , that is composed of the area-mean and areamedian diameters, W* and , calculated directly from the individual nozzle diameters for all nozzles on a given stage (equation 1): W= (W*)(2/3) x (W)(1/3) (1). Hence, the effective diameter provides an intuitive and technically sound basis for setting acceptance criteria for new and in-use impactors. We tabulate these criteria for the Mark II eight-stage Andersen cascade impactor and the Next Generation Pharmaceutical Impactor in a manner similar to the tables of critical impactor dimensions published in EP Supplement 5.1 and in USP 28. For two different impactors or for one impactor measured at two different times (e.g., at manufacture and in use), we find that the D50 values of a given stage are related to the effective diameters by D(50,2)/D(50,1)= (W(2)/W(1))(3/2) (2). Using the stage mensuration data for new, as-manufactured NGIs, we compare the D(50 )values of the first 125 as manufactured NGIs with those of the archivally calibrated NGI. We further establish that the archivally calibrated NGI has D(50) values within 0.3% of an entirely perfect, hypothetical NGI with all nozzles equal to the nominal nozzle diameters. We also apply the equations to a specific mensurated impactor to show that a used impactor with some nozzles outside of the original manufacturing specifications can have the same aerodynamic performance as a new impactor. PMID- 16379617 TI - Use of the next generation pharmaceutical impactor for particle size distribution measurements of live viral aerosol vaccines. AB - Aerosol administration of live measles vaccine virus has proven to be extremely efficacious in field trials using an industrial compressor coupled to a disposable nebulizer (IPI). To develop a new system for administration, it is necessary to characterize the operating characteristics of the old system. There are no standardized techniques for measuring particle size of live biological agents. This study evaluated the Next Generation Pharmaceutical Impactor's (NGI) ability to particle size wet aerosols in an effort to measure the particle size distribution of live measles vaccine from the IPI nebulizer. As a control albuterol was aerosolized using a Pari LC Star, since the soluble albuterol is evenly distributed throughout the droplets and laser diffraction measurements should agree with those from the NGI, as long as the NGI is cooled to prevent heat transfer to the aerosol. Albuterol was also used as a control for the IPI using quantitative ultraviolet spectrophotometry. There was close agreement in MMD (mean +/- 95% CI) for the LC Star, measured by laser diffraction (3.24 +/- 0.06 microm) and the NGI (2.93 +/- 0.22 microm) and the IPI (4.26 +/- 0.17 and 4.26 +/- 0.24 microm, respectively). For the measles vaccine assayed for plaque forming units, there were significant differences between the NGI MMD (6.14 +/- 0.39 microm) compared to laser diffraction (4.95 +/- 0.16 microm) indicating that the vaccine is not evenly distributed among the droplets of various sizes. This is likely clumping of the virus due to gelatin in the formulation. These data indicate that the NGI is capable of particle sizing live biological agents. PMID- 16379618 TI - In vitro estimations of in vivo jet nebulizer efficiency using actual and simulated tidal breathing patterns. AB - In vivo aerosol delivery efficiency was estimated in vitro for two jet nebulizers using a breath monitor (Breathe!; Pari GmbH, Germany) and breath simulator (COMPAS; Pari GmbH) to reproduce subject tidal breathing patterns. The AeroEclipse (Trudell Medical International, Canada), a breath-actuated nebulizer, and the LC Star (Pari GmbH), a breath-enhanced nebulizer, were filled with levalbuterol HCl solution (Sepracor, USA) and operated with compressed O(2) at 8 lpm. Tidal breathing patterns of 20 adult subjects were digitally recorded with the Breathe! Breath Monitor. Subjects then breathed tidally from each nebulizer separately for 1 minute and to nebulizer dryness. Levalbuterol aerosol collected on filters placed between the nebulizer and mouth was chemically assayed to determine the inspired mass (IM), wasted mass (WM) and total emitted mass (TM). Measurements were repeated using the COMPAS Breath Simulator to simulate each subject's tidal breathing pattern. IM, WM, and TM measurements using actual versus simulated tidal breathing were highly comparable for each nebulizer, except the IM (p < 0.05) from LC Star measured at nebulizer dryness. Breath simulation was an inaccurate tool for estimating the time to nebulizer dryness as simulated measurements to nebulizer dryness took significantly longer than measurements preformed with actual tidal breathing (p < 0.001). While breath simulation provides an accurate in vitro tool for estimating in vivo aerosol delivery, it should not completely replace in vivo measurements until inherent limitations in simulator operation can be overcome to provide a more clinically realistic simulation. PMID- 16379619 TI - Analysis of cascade impactor mass distributions. AB - The purpose of this paper is to review the approaches for analyzing cascade impactor (CI) mass distributions produced by pulmonary drug products and the considerations necessary for selecting the appropriate analysis procedure. There are several methods available for analyzing CI data, yielding a hierarchy of information in terms of nominal, ordinal and continuous variables. Mass distributions analyzed as a nominal function of the stages and auxiliary components is the simplest approach for examining the whole mass emitted by the inhaler. However, the relationship between the mass distribution and aerodynamic diameter is not described by such data. This relationship is a critical attribute of pulmonary drug products due to the association between aerodynamic diameter and the mass of particulates deposited to the respiratory tract. Therefore, the nominal mass distribution can only be utilized to make decisions on the discrete masses collected in the CI. Mass distributions analyzed as an ordinal function of aerodynamic diameter can be obtained by introducing the stage size range, which generally vary in magnitude from one stage to another for a given type of CI, and differ between CIs of different designs. Furthermore, the mass collected by specific size ranges within the CI are often incorrectly used to estimate in vivo deposition at various regions of the respiratory tract. A CI-generated mass distribution can be directly related to aerodynamic diameter by expressing the mass collected by each size-fractionating stage in terms of either mass frequency or cumulative mass fraction less than the aerodynamic size appropriate to each stage. Analysis of the aerodynamic diameter as a continuous variable allows comparison of mass distributions obtained from different products, obtained by different CI designs, as well as providing input to in vivo particle deposition models. The lack of information about the mass fraction emitted by the inhaler that is not size-analyzed by the CI may be perceived as a disadvantage from the standpoint of comparing the total mass per actuation emitted from the inhaler mouthpiece. However, this is a limitation of the CI measurement technique rather than the data analysis procedure. Data reduction techniques can enable the large quantity of information conveyed in a mass-size distribution to be summarized in terms of representative parameters, but care needs to be exercised if utilizing model size distribution function fitting routines to avoid introducing error by the fitting procedure. PMID- 16379620 TI - Dose delivery characteristics of the AIR pulmonary delivery system over a range of inspiratory flow rates. AB - The purpose of this study was to evaluate the in vitro and in vivo dose delivery characteristics of the AIR pulmonary delivery system over a range of flow rates. A 5-mg placebo powder of engineered particles with low densities (<0.4 g/cc) and large geometric diameters (>5 microm) was delivered via a simple, capsule based, passive dry powder inhaler. The emitted dose, geometric and aerodynamic particle size distributions (aPSDs) were obtained over a range of flow rates (15-60 LPM). The in vitro results demonstrated improved powder dispersion with increasing flow rate through the inhaler. The in vivo dose delivery characteristics were obtained by gamma scintigraphy. Twelve healthy subjects performed the following three inhalation maneuvers: (i) a targeted peak inspiratory flow rate (PIFR) of 20 +/- 10 LPM, (ii) a deep comfortable inhalation, and (iii) a deep forced inhalation. PIFR and inhaled volume were obtained during the inhalation of the dose using a spirometer. In vivo dose delivery was characterized by high and reproducible emitted doses (mean = 87%; inter and intra-subject CV = 5%) and high lung deposition (mean = 51% of the total dose), with low inter and intra-subject CVs (18% and 13%, respectively) across a range of PIFRs (12-86 LPM). Lung deposition of the total dose was shown not to be dependent on PIFR by analysis of variance across the range of inspiratory flow rates (p = 0.29). This was due to the competing effects of smaller aPSDs, increased extrathoracic deposition and higher emitted doses with increasing PIFR. Fully characterizing the effect of inspiratory flow rate requires analysis of the therapeutic response, as well as in vitro dose delivery and lung deposition. PMID- 16379621 TI - Experimental measurements of particle deposition in three proximal lung bifurcation models with an idealized mouth-throat. AB - In this paper, particle deposition in three idealized proximal lung bifurcation models with an idealized mouth-throat were investigated experimentally. These bifurcation models included (1) a small symmetric bifurcation, (2) an intermediate asymmetric bifurcation, and (3) a large symmetric bifurcation. An idealized mouth-throat geometry (the "Alberta geometry") was used as the inlet to these bifurcation models. Monodisperse aerosol particles of DEHS (di-2-ethylhexyl sebecate) oil with mass median diameters in the range of 2.5-7.5 microm were employed at steady flow rates of 30-90 L/min. Particle deposition measurements were conducted by gravimetry. The results show that particle deposition in the mouth-throat and trachea accounts for the major portion of total deposition in the entire models used, and particle deposition fraction in the proximal lung bifurcations is lower compared with that deposited in the regions upstream (the mouth-throat and the trachea). Total particle deposition efficiency increases with increasing either inertial parameter or Stokes number. Total particle deposition varies appreciably from model to model. The laryngeal jet is the key factor dominating particle deposition within the trachea. An effect of Reynolds number on particle deposition efficiency in the trachea is observed. In addition, particle deposition in the bifurcation region is influenced little by the upstream flow condition, and therefore the effect of the laryngeal jet on deposition seemingly does not propagate to the bifurcations downstream. PMID- 16379622 TI - What is AVMA's role in moral guidance? PMID- 16379623 TI - What is your diagnosis? Traumatic hernia of the body wall. PMID- 16379624 TI - Anesthesia case of the month. Severe twist in the inner tube of the coaxial breathing circuit, resulting in obstruction of the inspiratory limb of the circuit. PMID- 16379625 TI - Effects of veterinary board disciplinary actions on veterinarians licensed in multiple states. PMID- 16379626 TI - Rabies surveillance in the United States during 2004. AB - During 2004, 49 states and Puerto Rico reported 6,836 cases of rabies in nonhuman animals and 8 cases in human beings to the CDC, representing a 4.6% decrease from the 7,170 cases in nonhuman animals and 3 cases in human beings reported in 2003. Approximately 92% of the cases were in wildlife, and 8% were in domestic animals (compared with 91% and 9%, respectively, in 2003). Relative contributions by the major animal groups were as follows: 2,564 raccoons (37.5%), 1,856 skunks (27.1%), 1,361 bats (19.9%), 389 foxes (5.7%), 281 cats (4.1%), 115 cattle (1.7%), and 94 dogs (1.4%). Compared with the numbers of reported cases in 2003, cases in 2004 decreased among all groups, except bats, cattle, human beings, and "other domestics" (1 llama). Decreases in numbers of rabid raccoons during 2004 were reported by 12 of the 20 eastern states in which raccoon rabies was enzootic. In the East, Massachusetts reported the first cases of raccoon rabies detected beyond the Cape Cod oral rabies vaccine barrier. Along the western edge of the raccoon rabies epizootic (Ohio in the north and Tennessee in the south), cases of rabies were reported from unexpected new foci beyond oral rabies vaccine zones. On a national level, the number of rabies cases in skunks during 2004 decreased by 12.1% from the number reported in 2003. Once again, Texas reported the greatest number (n = 534) of rabid skunks and the greatest overall state total of rabies cases (913). Texas reported only 1 case of rabies in a dog that was infected with the dog/coyote rabies virus variant and only 22 cases associated with theTexas gray fox rabies virus variant (compared with 61 cases in 2003). The total number of cases of rabies reported nationally in foxes and raccoons declined 14.7% and 2.7%, respectively, during 2004. The 1,361 cases of rabies reported in bats during 2004 represented a 12.3% increase over the previous year's total of 1,212 cases for this group of mammals. Cases of rabies reported in cats, dogs, horses and mules, and sheep and goats decreased 12.5%, 19.7%, 31.8%, and 16.7%, respectively, whereas cases reported in cattle increased 174%. In Puerto Rico, reported cases of rabies in mongooses decreased 4.1% and rabies in dogs (9 cases) remained unchanged from those reported in 2003. Among the 8 cases of rabies in human beings, 1 person from Oklahoma and 3 from Texas died following receipt of infected organs and tissues from an Arkansas donor. In California, a person originally from El Salvador and, in Florida, a person originally from Haiti both died of canine rabies infections acquired outside the United States. In Wisconsin, a teenager contracted rabies from a bat bite and became the first known person to survive rabies despite not having received rabies vaccine prior to symptom onset. PMID- 16379629 TI - Owner impressions of three premium diets fed to healthy adult dogs. AB - OBJECTIVE: To determine owner impressions of 3 premium canine diets when factors such as price and retail source were removed; to compare body condition scores (BCSs) assigned by owners versus a veterinarian; and to determine consistency of owner impressions of diets when owners were not informed that they were feeding the same diet during 2 consecutive periods. DESIGN: Randomized controlled trial. ANIMALS: 44 healthy adult dogs. PROCEDURE: During the initial 12 months of the study, dogs were each fed 3 premium diets for 4 months in random order. After feeding each diet for 1 and 4 months, owners completed questionnaires regarding palatability of the diet; the dog's attitude, energy level, fecal consistency, frequency of defecation, hair coat quality, and BCS; and whether they would feed the diet if available commercially. During the last 4 months of the study, owners fed the same diet they had been feeding during the previous 4 months. RESULTS: Scores for most variables did not differ among diets. However, mean BCS assigned by owners was significantly lower than mean BCS assigned by an investigator, with a moderate correlation between scores. When asked at the end of the third and fourth study periods whether they would consider feeding the diet long-term, 12 of the 44 (27%) owners gave inconsistent responses. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that when unaware of retail price and source, owners have similar impressions of 3 premium diets fed to healthy adult dogs, suggesting that factors other than the diets themselves may affect owner impressions. Owners also underestimate their dog's BCS. PMID- 16379631 TI - Fecal incontinence and spinal cord abnormalities in seven dogs. AB - Seven dogs with fecal incontinence and abnormal gaits were evaluated. Fecal incontinence was characterized as defecation of normal stools without posturing. Duration of clinical signs prior to evaluation ranged from 5 months to 3 years. Five dogs had upper motor neuron (UMN) paraparesis, and 2 dogs had UMN tetraparesis. With magnetic resonance imaging, spinal cord abnormalities primarily involving the dorsal aspect of the spinal cord were identified in all dogs. Five dogs had focal abnormalities, and 2 dogs had diffuse abnormalities of the spinal cord. Of the dogs with focal spinal cord lesions, 4 had cystic spinal cord abnormalities and 1 had a meningioma. Surgery was performed on all dogs with focal lesions; 4 of the 5 dogs had resolution of fecal incontinence after surgery. Results in these dogs suggest that fecal incontinence can be associated with spinal cord abnormalities and, depending on the characteristics of the lesion, can resolve after surgical treatment of the abnormality. PMID- 16379630 TI - Comparison of oral and subcutaneous administration of buprenorphine and meloxicam for preemptive analgesia in cats undergoing ovariohysterectomy. AB - OBJECTIVE: To compare the effectiveness of preoperative PO and SC administration of buprenorphine and meloxicam for prevention of postoperative pain-associated behaviors in cats undergoing ovariohysterectomy. DESIGN: Randomized controlled study. ANIMALS: 51 female cats (4 to 60 months old; weight range, 1.41 to 4.73 kg [3.1 to 10.4 lb]). PROCEDURE: Cats received 1 of 5 treatments at the time of anesthetic induction: buprenorphine PO (0.01 mg/kg [0.0045 mg/lb]; n = 10), buprenorphine SC (0.01 mg/kg; 10), meloxicam SC (0.3 mg/kg 10.14 mg/lb]; 10), meloxicam PO (0.3 mg/kg; 10), or 0.3 mL of sterile saline (0.9% NaCI) solution SC (control group; 11). Sedation scores and visual analog scale and interactive visual analog scale (IVAS) pain-associated behavior scores were assigned to each cat 2 hours before and at intervals until 20 hours after surgery. RESULTS: Cats receiving meloxicam PO or SC had significantly lower IVAS scores (2.91 and 2.02, respectively), compared with IVAS scores for cats receiving buprenorphine PO (755). Pain-associated behavior scores for cats administered buprenorphine or meloxicam PO or SC preoperatively did not differ significantly from control group scores. Rescue analgesia was not required by any of the cats receiving meloxicam, whereas 3 of 10 cats receiving buprenorphine PO, 2 of 10 cats receiving buprenorphine SC, and 1 of 11 cats receiving the control treatment required rescue analgesia. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of pain associated behavior scores, cats receiving meloxicam PO or SC before ovariohysterectomy appeared to have less pain after surgery than those receiving buprenorphine PO preoperatively. PMID- 16379632 TI - Coil embolization of a congenital orbital varix in a dog. AB - A 10-week-old Labrador Retriever was examined because of a swelling above the left eye. Ophthalmic examination revealed a tubular, light-pink, slightly raised lesion of the left conjunctiva that extended from the limbus to the fornix and into the dorsal eyelid. The lesion affected the entire margin of the dorsal eyelid and extended 2 cm dorsal to the eyelid margin. With compression of the left jugular vein, the exophthalmos worsened immediately and the subconjunctival and eyelid lesion enlarged. Results of ultrasonography, computed tomography, and contrast venography were consistent with a diagnosis of an orbital varix. Coil embolization was elected for treatment of the varix to prevent the pain and morbidity associated with an orbitotomy. Coils were introduced through a 22-gauge IV catheter inserted through the upper eyelid into the varix. The only complication was moderately severe orbital swelling. The owners reported that the lesion had resolved by 2 weeks after coil embolization. PMID- 16379633 TI - Outcome following surgical removal of nonvisceral soft tissue sarcomas in cats: 42 cases (1992-2000). AB - OBJECTIVE: To identify factors associated with outcome of cats with nonvisceral soft tissue sarcomas treated with surgery alone. DESIGN: Retrospective study. ANIMALS: 42 cats. PROCEDURE: Medical records were reviewed for clinically relevant data, and histologic samples were examined. Follow-up information was obtained by means of physical examination or through telephone conversations with referring veterinarians and owners. The Kaplan-Meier method was used to construct survival curves. RESULTS: Median survival time was 608 days (range, 85 to 2,291 days), although 24 cats were still alive at the time of the study. Tumor size (ie, diameter) and histologic type were significantly associated with survival time. Median survival time was significantly longer in cats with tumors that were < 2 cm in diameter, compared with cats in which tumors were > 2 cm. Median survival times for cats with a fibrosarcoma or nerve sheath tumor were significantly longer than median time for cats with a malignant fibrous histiocytoma. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that tumor size and type are significantly associated with survival time in cats with nonvisceral soft tissue tumors. PMID- 16379634 TI - Prevalences and clinical signs of polysaccharide storage myopathy and shivers in Belgian draft horses. AB - OBJECTIVE: To determine prevalences of polysaccharide storage myopathy (PSSM) and shivers in Belgian Draft Horses (BDHs) and determine whether there was an association between these 2 conditions. DESIGN: Prospective cohort study. ANIMALS: 103 BDHs > 1 year old. PROCEDURE: Owners were questioned regarding clinical signs of PSSM, shivers, and hindquarter weakness, defined as poor hindquarter muscling and lack of propulsion. Blood samples were collected for determination of serum creatine kinase and aspartate transferase activities and serum selenium and vitamin E concentrations. A biopsy sample from the gluteus medius muscle was submitted for histologic, histochemical, and biochemical analysis. A diagnosis of PSSM was made if abnormal amylase-resistant polysaccharide inclusions were seen histologically. RESULTS: 37 (36%) horses had PSSM and 19 (18%) had shivers, but only 6 (6%) had both PSSM and shivers, whereas 31 (30%) had PSSM alone, 13 (13%) had shivers alone, and 53 (51%) had neither, and a significant association between PSSM and shivers was not detected. Hindquarter weakness was found in 30 horses. Only 13 of 37 (35%) horses with PSSM and 11 of 19 (58%) horses with shivers had hindquarter weakness. Serum creatine kinase and aspartate transferase activities and serum selenium and vitamin E concentrations were not significantly different between horses with and without PSSM or between horses with and without shivers. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that PSSM and shivers are common but unrelated disorders in BDHs. PMID- 16379635 TI - Rupture of the gastrocnemius muscle in six foals. AB - Rupture of the gastrocnemius muscle and subsequent disruption of the reciprocal mechanism of the hind limb was diagnosed in 6 foals examined at 7 hours to 3 weeks of age. In 2 foals, the musculoskeletal injury was detected as an ancillary finding to clinical signs of neurologic dysfunction ascribed to hypoxic ischemic insult during delivery, whereas in the other 4 foals, musculoskeletal injury, manifested as inability to rise or stand unsupported, was the chief complaint at admission. Five foals had a history of dystocia and assisted delivery. Common clinical signs were inability to rise, disruption of the reciprocal mechanism, swelling in the caudal aspect of the thigh, instability of the stifle joint, and stifle joint effusion. For mild gastrocnemius injury, exercise restriction via forced recumbency, with minimal or no bandaging, may be sufficient treatment. For more severe disruption of the muscle, limb stabilization via splinting and intensive nursing and monitoring are necessary. Four foals had important concurrent problems, including musculoskeletal deformations (joint contractures), hypoxic ischemic disease, and failure of passive transfer and associated problems (ie, sepsis, polyarthritis, and pneumonia). Moderate to severe gastrocnemius muscle injury is difficult to treat successfully, and the long-term prognosis for athletic function should be regarded as guarded. PMID- 16379636 TI - Effect of prepartum intramammary treatment with pirlimycin hydrochloride on prevalence of early first-lactation mastitis in dairy heifers. AB - OBJECTIVE: To determine whether prepartum intramammary treatment of dairy heifers with pirlimycin hydrochloride would reduce the prevalence of intramammary infection (IMI) and lower the somatic cell count (SCC) during early lactation or improve 305-day mature equivalent milk production. DESIGN: Prospective clinical trial. ANIMALS: 183 Holstein-Friesian heifers (663 quarters) from 2 dairy farms. PROCEDURE: Heifers were assigned to treatment and control groups. Treated heifers received a single 50-mg dose of pirlimycin in each mammary quarter approximately 10 to 14 days prior to parturition. Prepartum mammary gland secretions and postpartum milk samples were collected for bacterial culture. Postpartum milk samples were also collected for determination of SCC or California mastitis testing and were tested for pirlimycin residues. Mature equivalent 305-day milk production data were recorded. RESULTS: Treated heifers in herd A had a higher overall cure rate, higher cure rates for IMI caused by coagulase-negative staphylococci (CNS) and Staphylococcus aureus, lower SCC, and lower prevalence of chronic IMI, compared with control heifers. Treated heifers in herd B had a higher overall cure rate and cure rate for IMI caused by CNS, compared with control heifers, but postpartum California mastitis test scores and prevalence of chronic IMI did not differ between groups. Mature equivalent 305-day milk production did not differ between herds or treatment groups. No pirlimycin residues were detected in postpartum milk samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that prepartum treatment of dairy heifers with pirlimycin may reduce the prevalence of early lactation IMI, particularly IMI caused by CNS, without causing pirlimycin residues in milk. PMID- 16379637 TI - Risk of removal and effects on milk production associated with paratuberculosis status in dairy cows. AB - OBJECTIVE: To determine effects on production and risk of removal related to Mycobacterium avium subsp paratuberculosis (MAP) infection at the individual animal level in dairy cattle. DESIGN: Longitudinal study. ANIMALS: 7,879 dairy cows from 38 herds in 16 states. PROCEDURE: A subset of dairy cattle operations that participated in the National Animal Health Monitoring System Dairy 2002 study was evaluated via a serum ELISA for antibodies against MAP and categorized according to ELISA score. Dairy Herd Improvement Association records were obtained to collect current and historical lactation data and removal (ie, culling) information. Production variables were evaluated on the basis of serum ELISA category. RESULTS: Cows with strong positive results had mature equivalent (ME) 305-day milk production, ME 305-day maximum milk production, and total lifetime milk production that were significantly lower than cows in other categories. No differences were observed for ME 305-day fat and protein percentages, age, lactation, and lactation mean linear somatic cell count score between cows with strong positive results and those with negative results. After accounting for lactation number and relative herd-level milk production, cows with strong positive results were significantly more likely to have been removed by 1 year after testing. CONCLUSIONS AND CLINICAL RELEVANCE: Without management changes designed to reduce the farm-level prevalence of MAP infection, paratuberculosis will continue to reduce farm income by decreasing milk production and potentially increasing premature removal from the herd. PMID- 16379640 TI - Questions justification for inoculation of dogs. PMID- 16379641 TI - Effect of intensity of fecal pat sampling on estimates of Escherichia coli O157 prevalence. AB - OBJECTIVE: To evaluate site-to-site variation within fecal pats from cattle with regard to detection of Escherichia coli O157 and determine the effect on the accuracy of prevalence estimates of assay of multiple samples collected from the same fecal pat. SAMPLE POPULATION: 120 freshly voided fecal pats collected from 2 beef feedlots. Procedures-5 samples were systematically collected from each fecal pat and analyzed for E coli O157 via selective preenrichment techniques, immunomagnetic separation, and biochemical tests. Presumptive isolates were definitively identified via agglutination assays and polymerase chain reaction techniques. Best estimators of prevalence were calculated from the distribution of E coli O157-positive samples per pat. RESULTS: Of the 120 fecal pats, 96, 13, 4, 2, 3, and 2 fecal pats had 0, 1, 2, 3, 4, and 5 E coli O157-positive samples, respectively. The greatest estimate of E coli O157 prevalence (20%) was achieved when all 5 samples were assessed; this estimate represented a 2.4- fold increase in prevalence, compared with that provided via analysis of 1 sample/pat (8.2%). Compared with assessment of 5 sites/pat, the relative sensitivity of detecting an E coli O157-positive fecal pat via analysis of 1 site/pat was 40.1%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that estimates of E coli O157 prevalence derived from sampling of 1 location/pat are likely underestimates of the true prevalence of this pathogen in fecal pats (and by extension, cattle). Additional research is warranted to confirm these results in situations of high and low prevalence and across different feedlots. PMID- 16379642 TI - Direct and indirect markers of cartilage metabolism in synovial fluid obtained from dogs with hip dysplasia and correlation with clinical and radiographic variables. AB - OBJECTIVE: To compare activities of interleukin (IL)-1beta, IL-6, tumor necrosis factor (TNF)-alpha, and matrix metalloproteinase (MMP)-3 and contents of sulfated glycosaminoglycan (S-GAG) in joint fluid obtained from dogs with hip dysplasia (HD) and clinically normal dogs, evaluate correlations among these markers in joint fluid obtained from dogs with HD, and evaluate correlations between each marker and clinical and radiographic variables. Animals-26 dogs with HD (clinical group) and 43 clinically normal Beagles (control group). PROCEDURE: Joint fluid was aseptically collected from the hip joints of all dogs. For each dog in the clinical group, age, duration of lameness, radiographic osteoarthritis (OA) score, and Norberg angle in each affected joint were recorded. Activities of IL 1beta, IL-6, TNF-alpha, and MMP-3 and S-GAG contents were measured. Values were compared between groups by use of Mann-Whitney U tests, and the Spearman rank correlation test was used to evaluate correlations among markers and between each marker and clinical or radiographic variables. RESULTS: Values of all markers were significantly higher for the clinical group, compared with values for the control group. There was a moderate positive correlation between lameness duration and IL-6 activity and a strong negative correlation between the Norberg angle and IL-1beta activity. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of our results indicated that there was a significant increase in markers of OA in dogs with HD. Activities of IL-1beta and IL-6 in joint fluid of dogs with HD may be influenced by the severity of laxity in the hip joint and lameness duration, respectively. PMID- 16379643 TI - Pharmacokinetics of ketamine and its metabolite, norketamine, after intravenous administration of a bolus of ketamine to isoflurane-anesthetized dogs. AB - OBJECTIVE: To determine the pharmacokinetics of ketamine and norketamine in isoflurane-anesthetized dogs. Animals-6 dogs. PROCEDURE: The minimum alveolar concentration (MAC) of isoflurane was determined in each dog. Isoflurane concentration was then set at 0.75 times the individual's MAC, and ketamine (3 mg/kg) was administered IV. Blood samples were collected at various times following ketamine administration. Blood was immediately centrifuged, and the plasma separated and frozen until analyzed. Ketamine and norketamine concentrations were measured in the plasma samples by use of liquid chromatography-mass spectrometry. Ketamine concentration-time data were fitted to compartment models. Norketamine concentration-time data were examined by use of noncompartmental analysis. RESULTS: The MAC of isoflurane was 1.43 +/- 0.18% (mean +/- SD). A 2-compartment model best described the disposition of ketamine. The apparent volume of distribution of the central compartment, the apparent volume of distribution at steady state, and the clearance were 371.3 +/- 162 mL/kg, 4,060.3 +/- 2,405.7 mL/kg, and 58.2 +/- 17.3 mL/min/kg, respectively. Norketamine rapidly appeared in plasma following ketamine administration and had a terminal half-life of 63.6 +/- 23.9 minutes. A large variability in plasma concentrations, and therefore pharmacokinetic parameters, was observed among dogs for ketamine and norketamine. CONCLUSIONS AND CLINICAL RELEVANCE: In isofluraneanesthetized dogs, a high variability in the disposition of ketamine appears to exist among individuals. The disposition of ketamine may be difficult to predict in clinical patients. PMID- 16379644 TI - Evaluation of the internal vertebral venous plexus, vertebral canal, dural sac, and vertebral body via nonselective computed tomographic venography in the cervical vertebral column in healthy dogs. AB - OBJECTIVE: To evaluate nonselective computed tomographic (CT) venography for evaluating the cervical internal vertebral venous plexus (IVVP), define the diameter and area dimensions of the IVVP, and determine the relationship between dimensions of the cervical IVVP and other vertebral components in medium-sized dogs. Animals-6 healthy dogs that weighed 18 to 27 kg. Procedure-Helical CT scans were performed from C1 to C7 before and after IV injection of contrast medium (480 mg of iodine/kg) and a continuous infusion (240 mg of iodine/kg). Image data were transferred to a CT workstation, and measurements were performed on displayed transverse images. Diameter and area measurements of the vertebral canal, dural sac, IVVP, and vertebral body were obtained at C3 to C7. RESULTS: Opacification of vertebral venous structures was achieved in all dogs with no adverse reactions. Sagittal diameters of the IVVP for C3 to C7 ranged from 0.6 to 3.2 mm. Transverse diameters ranged from 2.32 to 5.74 mm. The IVVP area represented 12.4% of the mean vertebral canal transverse area and 30.61% of the mean vertebral epidural space area. Area measurements of the IVVP were significantly correlated with vertebral canal area and dural sac area. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that nonselective CT venography is a safe, sensitive method for performing morphometric assessments of the cervical IVVP in dogs. Findings support the theory that there may be a physiologic or developmental relationship between cervical vertebral canal components. PMID- 16379645 TI - Use of gyroscopic sensors for objective evaluation of trimming and shoeing to alter time between heel and toe lift-off at end of the stance phase in horses walking and trotting on a treadmill. AB - OBJECTIVE: To determine whether a shoe with an axialcontoured lateral branch would induce greater lateral roll of the forelimb hoof during the time between heel and toe lift-off at end of the stance phase (breakover). Animals-10 adult horses. PROCEDURE: A gyroscopic transducer was placed on the hoof of the right forelimb and connected to a transmitter. Data on hoof angular velocity were collected as each horse walked and trotted on a treadmill before (treatment 1, no trim-no shoe) and after 2 treatments by a farrier (treatment 2, trim-standard shoe; and treatment 3, trim-contoured shoe). Data were converted to hoof angles by mathematical integration. Breakover duration was divided into 4 segments, and hoof angles in 3 planes (pitch, roll, and yaw) were calculated at the end of each segment. Multivariable ANOVA was performed to detect differences among treatments and gaits. RESULTS: Trimming and shoeing with a shoe with contoured lateral branches induced greater mean lateral roll to the hoof of 3.2 degrees and 2.5 degrees during the first half of breakover when trotting, compared with values for no trim-no shoe and trim-standard shoe, respectively. This effect dissipated during the second half of breakover. When horses walked, lateral roll during breakover was not significantly enhanced by use of this shoe. CONCLUSIONS AND CLINICAL RELEVANCE: A shoe with an axial-contoured lateral branch induced greater lateral roll during breakover in trotting horses, but change in orientation of the hoof was small and limited to the first half of breakover. PMID- 16379646 TI - Agreement in histologic assessments of the pituitary pars intermedia in aged horses. AB - OBJECTIVE: To evaluate concordance among veterinary pathologists in the assessment of histologic findings in the pars intermedia of pituitary gland sections from aged horses with mild signs suggestive of pituitary pars intermedia dysfunction (PPID). Sample Population-10 pituitary glands from aged horses. PROCEDURE: 7 pathologists were provided with signalment, clinical signs, and a single H&E-stained pituitary gland section from 10 aged horses with mild signs suggestive of PPID. Pathologists described histologic findings for each section and stated whether findings were consistent with PPID. Agreement among pathologists and with antemortem diagnostic test results was calculated. RESULTS: Overall, only fair agreement was found among the pathologists as to which horses had histologic findings consistent with disease (mean +/- SE kappa value, 0.34 +/ 0.069). Interpretation of individual sections varied, with minimal agreement (4 or 5/7 pathologists) for 5 of 10 sections evaluated. Postmortem assessment was in agreement with an antemortem endocrine diagnostic test result 79% of the time. CONCLUSIONS AND CLINICAL RELEVANCE: Validation of antemortem diagnostic testing for PPID in horses often relies on the results of postmortem histologic evaluation. The lack of consensus in histologic interpretation of pituitary glands from aged horses with mild clinical signs in our study indicates that postmortem histologic evaluation of pituitary glands is an inappropriate standard in validation of antemortem diagnostic tests for detection of early PPID. Caution should be used when interpreting diagnostic test results in horses in which early PPID is suspected. PMID- 16379647 TI - Clinicopathologic findings in dogs seroreactive to Bartonella henselae antigens. AB - OBJECTIVE: To assess the potential clinical relevance of seroreactivity to Bartonella henselae antigens in dogs. ANIMALS: 40 dogs seroreactive to B henselae and 45 dogs that did not seroreact to B henselae. PROCEDURE: A case-control study was conducted. Clinical and clinicopathologic findings were extracted from medical records of each dog. RESULTS: Statistical differences were not detected between dogs seroreactive or nonseroreactive to B henselae when analyzed on the basis of disease category or results of hematologic, biochemical, urine, or cytologic analysis. However, seroreactivity to B henselae antigens was detected in 2 of 4 dogs with a clinical diagnosis of granulomatous meningoencephalitis, 3 of 4 dogs with immune-mediated hemolytic anemia, 3 of 4 dogs with infective endocarditis, 2 of 3 dogs with lymphoid neoplasia, and 5 of 10 dogs with polyarthritis. Additionally, seroreactivity to B henselae antigens was detected in 18 of 34 thrombocytopenic dogs and 14 of 27 dogs with neutrophilia. CONCLUSIONS AND CLINICAL RELEVANCE: Significant associations were not detected between seroreactivity to B henselae and various diseases. Prospective epidemiologic studies investigating specific diseases, such as meningoencephalitis or polyarthritis, and specific hematologic abnormalities, such as immunemediated hemolytic anemia or thrombocytopenia, should be conducted to further define the potential clinical relevance of antibodies against B henselae in dogs. IMPACT FOR HUMAN MEDICINE: Bartonella organisms are increasingly reported as pathogens that induce are increasingly reported as pathogens that induce chronic infections in humans and dogs. Dogs may serve as natural candidates for future study of the disease in humans. PMID- 16379648 TI - Systemic and pituitary pars intermedia antioxidant capacity associated with pars intermedia oxidative stress and dysfunction in horses. AB - OBJECTIVE: To determine whether a deficiency in systemic or local (pars intermedia) antioxidant capacity is associated with pituitary pars intermedia oxidative stress and pituitary pars intermedia dysfunction (PPID) in horses. SAMPLE POPULATION: Blood samples from 20 horses with PPID and 20 healthy client owned horses, archived paraffin-embedded adrenal gland and substantia nigra tissues from 20 horses, and pituitary gland tissue from 16 horses. PROCEDURES: Total glutathione, superoxide dismutase, and glutathione peroxidase activities were determined in RBCs. Accumulation of a systemic marker of oxidative stress (3 nitrotyrosine) was assessed in plasma and formalin-fixed, paraffin-embedded adrenal gland and substantia nigra tissues. Local antioxidants (total and manganese superoxide dismutase, glutathione peroxidase, and total glutathione) were measured in pars intermedia tissues. RESULTS: No significant differences existed in systemic antioxidant enzyme activity or accumulation of 3 nitrotyrosine between horses with PPID and control horses. In pituitary gland tissues, glutathione peroxidase activity was increased in horses with oxidative stress, whereas total glutathione concentration and superoxide dismutase activity remained unchanged. There was an age-associated decrease in manganese superoxide dismutase activity in the pars intermedia. CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence of systemic accumulation of oxidative stress markers or deficiencies in antioxidant capacity in horses with PPID, suggesting that these are unlikely to be major predisposing factors in the development of PPID. Manganese superoxide dismutase activity in the pars intermedia decreased significantly with increasing age. Role of an age-associated decrease in antioxidant capacity for the pars intermedia in the development of PPID in horses warrants further investigation. PMID- 16379649 TI - Inflammatory changes in ruptured canine cranial and human anterior cruciate ligaments. AB - OBJECTIVE: To compare expression of tartrate-resistant acid phosphatase (TRAP) and cathepsin K and histologic changes in canine cranial cruciate ligaments (CCLs) and human anterior cruciate ligaments (ACLs). STUDY POPULATION: Sections of cruciate ligaments from 15 dogs with ruptured CCLs, 8 aged dogs with intact CCLs, 14 human beings with ruptured ACLs, and 11 aged human beings with intact ACLs. PROCEDURE: The CCLs and ACLs were evaluated histologically, and cells containing TRAP and cathepsin K were identified histochemically and immunohistochemically, respectively. RESULTS: The proportion of ruptured CCLs that contained TRAP+ cells was significantly higher than the proportion of intact ACLs that did but similar to proportions of intact CCLs and ruptured ACLs that did. The proportion of ruptured CCLs that contained cathepsin K+ cells was significantly increased, compared with all other groups. Numbers of TRAP+ and cathepsin K+ cells were significantly increased in ruptured CCLs, compared with intact ACLs. The presence of TRAP+ cells was correlated with inflammatory changes, which were most prominent in ruptured CCLs. CONCLUSION AND CLINICAL RELEVANCE: Results suggest that synovial macrophage-like cells that produce TRAP are an important feature of the inflammation associated with CCL rupture in dogs. Identification of TRAP and cathepsin K in intact CCLs and ACLs from aged dogs suggests that these enzymes have a functional role in cruciate ligament remodeling and repair. We hypothesize that recruitment and activation of TRAP+ macrophage-like cells into the stifle joint synovium and CCL epiligament are critical features of the inflammatory arthritis that promotes progressive degradation and eventual rupture of the CCL in dogs. PMID- 16379650 TI - Effects of long-term dietary supplementation with clinoptilolite on incidence of parturient paresis and serum concentrations of total calcium, phosphate, magnesium, potassium, and sodium in dairy cows. AB - OBJECTIVE: To determine whether dietary supplementation with clinoptilolite affects the incidence of parturient paresis and serum concentrations of total calcium (tCa), inorganic phosphorus (PO(4) (2)), magnesium (Mg2+), potassium (K+), and sodium (Na+) in dairy cattle. ANIMALS: 52 dairy cows. Procedure-Cows were placed into 3 groups. The first 2 groups (group A [n = 17] and group B [17]) were offered a concentrate supplemented with 1.25% and 2.5% clinoptilolite, respectively. The third (group C [n = 18]) served as a control and was offered the concentrate alone. The experiment started 1 month before parturition and lasted until the beginning of the next nonlactating period. Around the time of calving, all cows were monitored for the development of parturient paresis. Blood samples were taken at the commencement of the experiment, on the day of calving, and thereafter at monthly intervals to measure serum tCa, PO(4) (2), Mg2+, K+, and Na+ concentrations. Results-The incidence of parturient paresis in group B cows was significantly lower, compared with group C cows. However, serum concentrations of tCa, P(O4) (2), Mg2+, K+, and Na+ were not significantly affected by long-term supplementation with clinoptilolite. CONCLUSIONS AND CLINICAL RELEVANCE: In the context of this experiment, clinoptilolite supplementation at 2.5% appeared to have reduced the incidence of parturient paresis in dairy cows, suggesting that its effectiveness depends on the amount incorporated in the ration of cows. Addition of clinoptilolite in the concentrate of dairy cows during the nonlactating period could be used as a cost-effective preventive treatment for parturient paresis. PMID- 16379651 TI - Assessment of plasma brain natriuretic peptide concentration in Boxers with arrhythmogenic right ventricular cardiomyopathy. AB - OBJECTIVE: To determine whether Boxers with a clinical diagnosis of arrhythmogenic right ventricular cardiomyopathy (ARVC) have increased plasma concentrations of brain natriuretic peptide (BNP), compared with concentrations in clinically normal dogs. ANIMALS: 13 Boxers with ARVC, 9 clinically normal Boxers, 10 clinically normal non-Boxer dogs, and 5 hound dogs with systolic dysfunction. PROCEDURE: All Boxers were evaluated via 24-hour ambulatory electrocardiography and echocardiography; the number of ventricular premature contractions (VPCs) per 24 hours was assessed. Hound dogs with cardiac pacing induced systolic dysfunction (positive control dogs) and clinically normal non Boxer dogs (negative control dogs) were evaluated echocardiographically. Three milliliters of blood was collected from each dog for measurement of plasma BNP concentration by use of a radioimmunoassay. RESULTS: Mean +/- SD plasma BNP concentration for the ARVC-affected Boxers, clinically normal Boxers, negative control dogs, and positive control dogs was 11.0 +/- 4.6 pg/mL, 7.9 +/- 3.2 pg/mL, 11.5 +/- 4.9 pg/mL, and 100.8 +/- 56.8 pg/mL, respectively. Compared with findings in the positive control group, plasma BNP concentration in each of the other 3 groups was significantly different. There was no significant difference in BNP concentration between the 2 groups of Boxers. A significant correlation between plasma BNP concentration and number of VPCs per 24 hours in the ARVC affected Boxers was not identified. CONCLUSIONS AND CLINICAL RELEVANCE: A significant difference in BNP concentration between Boxers with ARVC and clinically normal Boxers was not identified. Results suggest that BNP concentration may not be an indicator of ARVC in Boxers. PMID- 16379652 TI - Effect of marbofloxacin on cardiovascular variables in healthy isoflurane anesthetized dogs. AB - OBJECTIVE: To study the hemodynamic effects of marbofloxacin (MBF) in isoflurane anesthetized dogs. ANIMALS: 6 healthy 8-month-old Beagles. PROCEDURE: Anesthesia was induced with sodium thiopental and maintained with isoflurane. Cardiovascular variables were monitored throughout anesthesia. Marbofloxacin was administered by an IV bolus at 2 mg/kg, followed 10 minutes later by an infusion at a rate of 40 mg/kg/h for 30 minutes (total dose, 20 mg/kg). Plasma MBF concentrations were measured by high-performance liquid chromatography. RESULTS: The mean peak concentration during MBF infusion was 34.2 +/- 6.4 microg/mL. The IV administration of the MBF bolus did not alter any cardiovascular variable in isoflurane-anesthetized dogs. Significant changes were found during infusion when a cumulative dose of 12 mg/kg had been given. The maximal decreases observed at the end of the infusion were 16% in heart rate, 26% in systolic left ventricular pressure, 33% in systolic aortic pressure, 38% in diastolic aortic pressure, 29% in cardiac output, and 12% in QT interval. All dogs recovered rapidly from anesthesia at the end of the experiment. CONCLUSIONS AND CLINICAL RELEVANCE: MBF may safely be used at 2 mg/kg IV in isoflurane-anesthetized dogs, and significant adverse cardiovascular effects are found only when 6 to 8 times the recommended dose is given. PMID- 16379653 TI - Evaluation of skin sensitivity after shock wave treatment in horses. AB - OBJECTIVE: To evaluate the effects of shock wave treatment on cutaneous nerve function, compared with the effects of local nerve block and sedation. ANIMALS: 18 clinically sound Swiss Warmbloods. PROCEDURE: Horses were randomly allocated to 3 groups and received different amounts and types of shock waves (extracorporeal shock wave treatment [ESWT] or radial pressure wave treatment [RPWT]). Horses were sedated with xylazine and levomethadone. Shock waves were applied to the lateral palmar digital nerve at the level of the proximal sesamoid bones on 1 forelimb. Skin sensitivity was evaluated by means of an electrical stimulus at the coronary band before and 5 minutes after sedation and at 4, 24, and 48 hours after application of ESWT or RPWT. On the contralateral forelimb, skin sensitivity was tested before and 10 minutes after an abaxial sesamoid nerve block. RESULTS: No significant changes in skin sensitivity were detected, regardless of the shock wave protocol applied. Mean reaction thresholds after sedation were more than twice the baseline thresholds. After the abaxial sesamoid block, no reaction was recorded in any of the horses. CONCLUSIONS AND CLINICAL RELEVANCE: Application of ESWT or RPWT to the palmar digital nerve had no effect on cutaneous sensation distal to the treated region for at least 2 days after application. The analgesic effect of sedation on reaction to electrical stimuli was distinct but varied among horses. PMID- 16379654 TI - Gelatinolytic matrix metalloproteinases-2 and -9 in tracheobronchial lavage fluid obtained from calves with concurrent infections of Pasteurella multocida and Mycoplasma bovirhinis. AB - OBJECTIVE: To study gelatinolytic matrix metalloproteinases (MMPs) in tracheobronchial lavage fluid (TBLF) obtained from clinically normal calves and calves with Pasteurella multocida infection. SAMPLE POPULATION: Samples of TBLF obtained from 11 calves with clinical signs of respiratory tract disease and growth of P multocida and Mycoplasma spp during culture of TBLF and samples of TBLF from 6 clinically normal calves with no bacterial growth during culture of TBLF. PROCEDURE: MMPs in TBLF were analyzed by use of gelatin zymography. Gelatinases were identified on the basis of molecular weights and inhibition by EDTA. RESULTS: The main gelatinolytic MMPs detected were the proform (90 to 110 kd) and active form (75 to 85 kd) of MMP-9 (gelatinase B) and the proform (67 to 75 kd) and active form (< 65 kd) of MMP-2 (gelatinase A). Increased amounts of active MMP-2 and MMP-9 were detected in TBLF of calves with respiratory tract disease, compared with amounts of active MMP-2 and MMP-9 in TBLF of clinically normal calves. Concurrent infection with Mycoplasma bovirhinis in calves with pneumonia attributable to P multocida was associated with higher concentrations of MMP-9. CONCLUSIONS AND CLINICAL RELEVANCE: The host response to P multocida includes increases in MMP-2 and MMP-9 concentrations in TBLF. Greater amounts of MMPs detected in calves with concurrent M bovirhinis and P multocida infection indicates synergism between these organisms. PMID- 16379655 TI - Evaluation of safety and pharmacokinetics of vancomycin after intravenous regional limb perfusion in horses. AB - OBJECTIVE: To evaluate clinical variables, regional concentrations, and pharmacokinetics of vancomycin in the synovial fluid of distal forelimb joints of horses after IV regional limb perfusion. ANIMALS: 6 horses. PROCEDURE: Vancomycin was administered via IV regional limb perfusion to the distal portion of the forelimbs of anesthetized horses. Drug (300 mg of vancomycin hydrochloride in 60 mL of saline [0.9% NaCl] solution) was infused into 1 forelimb, whereas the contralateral limb served as a control and was perfused with 60 mL of saline solution. Solutions were injected into the lateral digital vein after digital exsanguination. Synovial fluid from the metacarpophalangeal (MTCP) and distal interphalangeal (DIP) joints and systemic blood were collected prior to perfusion and 15, 30, 45, 65, and 90 minutes after initiation of the infusion. Synovial fluid from the MTCP joint and blood were also obtained at 4, 8, 12, and 24 hours after infusion. Plasma urea and creatinine concentrations, degree of lameness, and certain clinical variables involving the MTCP joint and infusion site were assessed for 7 days. Results were compared between the vancomycin treatment and control groups. RESULTS: No complications or significant differences in renal function, lameness, or clinical variables were observed between groups. Vancomycin concentrations exceeded 4 microg/mL in MTCP joints for approximately 20 hours. Higher concentrations were reached in DIP joints than in MTCP joints. CONCLUSIONS AND CLINICAL RELEVANCE: IV regional limb perfusion with 300 mg of vancomycin as a 0.5% solution was safe and may be useful in horses as treatment for distal limb infections. PMID- 16379656 TI - Use of proxies and reference quintiles obtained from minimal model analysis for determination of insulin sensitivity and pancreatic beta-cell responsiveness in horses. AB - OBJECTIVE: To develop proxies calculated from basal plasma glucose and insulin concentrations that predict insulin sensitivity (SI; L.min(-1) x mU(-1)) and beta cell responsiveness (ie, acute insulin response to glucose [AIRg]; mU/L x min( 1)) and to determine reference quintiles for these and minimal model variables. ANIMALS: 1 laminitic pony and 46 healthy horses. PROCEDURE: Basal plasma glucose (mg/dL) and insulin (mU/L) concentrations were determined from blood samples obtained between 8:00 AM and 9:00 AM. Minimal model results for 46 horses were compared by equivalence testing with proxies for screening SI and pancreatic beta cell responsiveness in humans and with 2 new proxies for screening in horses (ie, reciprocal of the square root of insulin [RISQI] and modified insulin-to-glucose ratio [MIRG]). RESULTS: Best predictors of SI and AIRg were RISQI (r = 0.77) and MIRG (r = 0.75) as follows: SI = 7.93(RISQI) - 1.03 and AIRg = 70.1(MIRG) - 13.8, where RISQI equals plasma insulin concentration(-0.5) and MIRG equals [800 - 0.30(plasma insulin concentration 50)(2)]/(plasma glucose concentration - 30). Total predictive powers were 78% and 80% for RISQI and MIRG, respectively. Reference ranges and quintiles for a population of healthy horses were calculated nonparametrically. CONCLUSIONS AND CLINICAL RELEVANCE: Proxies for screening SI and pancreatic beta-cell responsiveness in horses from this study compared favorably with proxies used effectively for humans. Combined use of RISQI and MIRG will enable differentiation between compensated and uncompensated insulin resistance. The sample size of our study allowed for determination of sound reference range values and quintiles for healthy horses. PMID- 16379657 TI - Cardiovascular and respiratory effects of ketamine infusions in isoflurane anesthetized dogs before and during noxious stimulation. AB - OBJECTIVE: To characterize the effects of ketamine administration on the cardiovascular and respiratory systems and on acid-base balance and to record adverse effects of ketamine in isoflurane-anesthetized dogs. ANIMALS: 6 healthy adult mongrel dogs. PROCEDURE: Dogs were anesthetized with isoflurane (1.25 times the individual minimum alveolar concentration) in oxygen, and ketamine was administered IV to target pseudo-steady-state plasma concentrations of 0, 0.5, 1, 2, 5, 8, and 11 microg/mL. Isoflurane concentration was reduced to an equipotent concentration. Cardiovascular, respiratory, and acid-base variables; body temperature; urine production; and adverse effects were recorded before and during noxious stimulation. Cardiac index, stroke index, rate-pressure product, systemic vascular resistance index, pulmonary vascular resistance index, left ventricular stroke work index, right ventricular stroke work index, arterial oxygen concentration, mixed-venous oxygen concentration, oxygen delivery, oxygen consumption, oxygen extraction ratio, alveolar-arterial oxygen partial pressure gradient, and venous admixture were calculated. Plasma ketamine and norketamine concentrations were measured. RESULTS: Overall, ketamine administration improved ventilation, oxygenation, hemodynamics, and oxygen delivery in isoflurane anesthetized dogs in a dosedependent manner. With the addition of ketamine, core body temperature was maintained or increased and urine production was maintained at an acceptable amount. However, at the higher plasma ketamine concentrations, adverse effects such as spontaneous movement and profuse salivation were observed. Myoclonus and dysphoria were observed during recovery in most dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Infusion of ketamine appears to be a suitable technique for balanced anesthesia with isoflurane in dogs. Plasma ketamine concentrations between 2 to 3 microg/mL elicited the most benefits with minimal adverse effects. PMID- 16379658 TI - Association between the existence of calves persistently infected with bovine viral diarrhea virus and commingling on pen morbidity in feedlot cattle. AB - OBJECTIVE: To determine the association between the existence of a calf persistently infected (PI) with bovine viral diarrhea virus (BVDV) and pen morbidity. ANIMALS: 5,041 calves in 50 pens at a feedlot in Iowa. PROCEDURE: In a longitudinal study, ear notches were collected from cattle and tested for BVDV antigen. Characteristics of each pen (owner, sex, disease rate, number of groups, and source) were recorded. The association between the existence of a BVDV-PI calf and morbidity in each pen was examined. RESULTS: Commingling was associated with an increase in respiratory tract disease (odds ratio [OR], 3; 95% confidence interval [CI], 2.5 to 3.6). Ten BVDV-PI calves (10/5,041 [0.2%]) were identified in 8 of 50 pens. A BVDV-PI calf was associated with reduced pen-level respiratory tract disease (OR, 0.7; 95% CI, 0.5 to 0.9). Disease prevalence (mean +/- SD morbidity, 7.9 +/- 3.1%) was lowest in pens containing single-source cattle and a BVDV-PI calf (4 pens containing 302 cattle), compared with single-source cattle with no BVDV-PI calf (mean morbidity, 11.89 +/- 9.7%; 31 pens containing 3,093 cattle), commingled cattle with no BVDV-PI calf (mean morbidity, 29.3 +/- 16.22%; 11 pens containing 1,127 cattle), and commingled cattle with a BVDV-PI calf (mean morbidity, 28.6 +/- 10.1%; 4 pens containing 519 cattle). CONCLUSIONS AND CLINICAL RELEVANCE: Commingling was the greatest risk factor associated with morbidity in each pen. A BVDV-PI calf in a pen was not associated with increased disease prevalence in commingled groups. PMID- 16379659 TI - Comparison of the cardiopulmonary effects of anesthesia maintained by continuous infusion of ketamine and propofol with anesthesia maintained by inhalation of sevoflurane in goats undergoing magnetic resonance imaging. AB - OBJECTIVE: To compare the cardiopulmonary effects of anesthesia maintained by continuous infusion of ketamine and propofol with anesthesia maintained by inhalation of sevoflurane in goats undergoing magnetic resonance imaging. ANIMALS: 8 Saanen goats. PROCEDURES: Goats were anesthetized twice (1-month interval) following sedation with midazolam (0.4 mg/kg, IV). Anesthesia was induced via IV administration of ketamine (3 mg/kg) and propofol (1 mg/kg) and maintained with an IV infusion of ketamine (0.03 mg/kg/min) and propofol (0.3 mg/kg/min) and 100% inspired oxygen (K-P treatment) or induced via IV administration of propofol (4 mg/kg) and maintained via inhalation of sevoflurane in oxygen (end-expired concentration, 2.3%; 1X minimum alveolar concentration; SEVO treatment). Cardiopulmonary and blood gas variables were assessed at intervals after induction of anesthesia. RESULTS: Mean +/- SD end-expired sevoflurane was 2.24 +/- 0.2%; ketamine and propofol were infused at rates of 0.03 +/- 0.002 mg/kg/min and 0.29 +/- 0.02 mg/kg/min, respectively. Overall, administration of ketamine and propofol for total IV anesthesia was associated with a degree of immobility and effects on cardiopulmonary parameters that were comparable to those associated with anesthesia maintained by inhalation of sevoflurane. Compared with the K-P treatment group, mean and diastolic blood pressure values in the SEVO treatment group were significantly lower at most or all time points after induction of anesthesia. After both treatments, recovery from anesthesia was good or excellent. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that ketamine-propofol total IV anesthesia in goats breathing 100% oxygen is practical and safe for performance of magnetic resonance imaging procedures. PMID- 16379660 TI - Evaluation of the respiratory elimination kinetics of selenium after oral administration in sheep. AB - OBJECTIVE: To evaluate the respiratory excretion and elimination kinetics of organic and inorganic selenium after oral administration in sheep. ANIMALS: 38 crossbred sheep. PROCEDURES: Selenium was administered PO to sheep as a single dose of 0, 1, 2, 3, or 4 mg/kg as sodium selenite or selenomethionine. Expired air was collected and analyzed from all sheep at 4, 8, and 16 hours after administration. RESULTS: Clinical signs consistent with selenium intoxication were seen in treatment groups given sodium selenite but not in treatment groups given the equivalent amount of selenium as selenomethionine. However, a distinct garlic-like odor was evident in the breath of all sheep receiving 2 to 4 mg of selenium/kg. The intensity of odor in the breath did not correlate with clinical signs in affected animals receiving sodium selenite treatment. CONCLUSIONS AND CLINICAL RELEVANCE: The concentration of selenium in expired air was greater in sheep receiving selenium as selenomethionine than sodium selenite. The concentration of selenium in expired air from sheep receiving high doses of selenium (3 and 4 mg of selenium/kg) was larger and selenium was expired for a longer duration than the concentration of selenium in expired air from sheep receiving low doses of selenium (1 and 2 mg of selenium/kg). PMID- 16379661 TI - Evaluation of surveillance and sample collection methods to document freedom from infectious bovine rhinotracheitis in cattle populations. AB - OBJECTIVES: To assess the sensitivity of the current surveillance program used in Denmark for detecting outbreaks of infectious bovine rhinotracheitis (IBR) at the herd level and to evaluate the impact of alternative sample collection strategies on the sensitivity of the system in an acceptable time frame. SAMPLE POPULATION: Data from the Danish Central Husbandry Register on cattle of 24,355 and 25,233 beef herds and on 13,034 and 12,003 dairy herds in the years 2000 and 2001, respectively. PROCEDURES: Surveillance programs were evaluated under current sample collection conditions and under 3 alternative scenarios by use of simulation modeling. Data from the current detection component of the surveillance system were used as input, taking into consideration the sensitivity and specificity of bulktank milk and serologic testing. RESULTS: The current system identifies infected dairy herds within a 3-month period with desired accuracy largely because of the test characteristics and number of bulk-tank milk samples. The system is less likely to detect infected beef herds in a timely manner because surveillance in beef herds depends solely on serologic testing at the time of slaughter. The efficiency of surveillance in dairy cattle herds was not decreased substantially when the slaughter-surveillance component was omitted. CONCLUSIONS AND CLINICAL RELEVANCE: Geographically targeted sample collection during the high-risk season (winter) was predicted to increase the probability of rapid detection of IBR infection in cattle. This approach can be used for assessing other surveillance systems to determine the best strategies for detection of infected herds. PMID- 16379662 TI - Application of a scaling model to establish and validate an interval level pain scale for assessment of acute pain in dogs. AB - OBJECTIVE: To establish interval level measurement in a prototype composite measure pain scale (CMPS) for assessment of acute pain in dogs and to investigate the scale's validity. ANIMALS: 20 clinically normal dogs, 20 dogs with medical conditions, and 117 dogs undergoing surgery. PROCEDURE: First, a scaling model was applied to the CMPS descriptors to establish weights for each and create a continuous scale. Subsequently, 5 observers independently used the scale to score signs of pain in 4 groups of dogs (control dogs, dogs with medical conditions, and 40 dogs undergoing soft tissue or orthopedic surgery). Scores from each group and from groups of conditions perceived to cause no, mild, moderate, and severe pain were compared. In addition, the scale was applied to 77 dogs undergoing orthopedic or soft tissue surgery and scores were compared with simultaneously derived numeric rating scale (NRS) scores; comparisons were made between surgical groups and with time after surgery. RESULTS: Calculated scale descriptor weights ranged from -2.0 to 2.0 and were transformed to create a continuous scale from 0 to 10. Median CMPS scores differed significantly among the 4 study groups and among pain severity groups and were typically greater with increasing perceived pain severity. Agreement was determined between CMPS and NRS scores, and there was a significant and expected time effect and difference between the CMPS scores of dogs undergoing orthopedic and soft tissue surgery. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that this interval level measurement scale is a valid measure of acute pain in dogs. PMID- 16379663 TI - Variation in the N-terminal region of an M-like protein of Streptococcus equi and evaluation of its potential as a tool in epidemiologic studies. AB - OBJECTIVE: To develop a method for typing Streptococcus equi on the basis of the DNA sequence of the genes that produce an M-like protein and to compare isolates among the United States, Japan, and other countries. SAMPLE POPULATION: S equi strains CF32, Hidaka/95/2, and NCTC9682 as well as 82 other isolates from the United States, Japan, and other countries obtained during 1975 to 2001. PROCEDURE: DNA sequences of the structural genes ( SeM and SzPSe) that produce M like proteins were determined for 3 representative strains to find a variable region. Variability in this region of SeM was then determined for the other isolates. Amino acid sequences were deduced and analyzed phylogenetically by use of the neighbor-joining method. RESULTS: Sequence diversity was detected in the N terminal region of SeM but not in SzPSe of the 3 representative strains. Base substitutions in the variable region of SeM varied in a nonsynonymous manner, resulting in variation in the amino acid sequence. Eighty-five isolates were categorized as 32 types of SeM on the basis of differences in the deduced amino acid sequences. CONCLUSIONS AND CLINICAL RELEVANCE: This study documented a region in the N-terminal portion of SeM that varies in a nonsynonymous manner. This information should be useful in molecular epidemiologic studies of S equi. PMID- 16379664 TI - Pharmacokinetics of carvedilol after intravenous and oral administration in conscious healthy dogs. AB - OBJECTIVE: To determine the pharmacokinetics of carvedilol administered IV and orally and determine the dose of carvedilol required to maintain plasma concentrations associated with anticipated therapeutic efficacy when administered orally to dogs. ANIMALS: 8 healthy dogs. PROCEDURES: Blood samples were collected for 24 hours after single doses of carvedilol were administered IV (175 microg/kg) or PO (1.5 mg/kg) by use of a crossover nonrandomized design. Carvedilol concentrations were detected in plasma by use of high-performance liquid chromatography. Plasma drug concentration versus time curves were subjected to noncompartmental pharmacokinetic analysis. RESULTS: The median peak concentration (extrapolated) of carvedilol after IV administration was 476 ng/mL (range, 203 to 1,920 ng/mL), elimination half-life (t(1/2)) was 282 minutes (range, 19 to 1,021 minutes), and mean residence time (MRT) was 360 minutes (range, 19 to 819 minutes). Volume of distribution at steady state was 2.0 L/kg (range, 0.7 to 4.3 L/kg). After oral administration of carvedilol, the median peak concentration was 24 microg/mL (range, 9 to 173 microg/mL), time to maximum concentration was 90 minutes (range, 60 to 180 minutes), t(1/2) was 82 minutes (range, 64 to 138 minutes), and MRT was 182 minutes (range, 112 to 254 minutes). Median bioavailability after oral administration of carvedilol was 2.1% (range, 0.4% to 54%). CONCLUSIONS AND CLINICAL RELEVANCE: Although results suggested a 3 hour dosing interval on the basis of MRT, pharmacodynamic studies investigating the duration of beta-adrenoreceptor blockade provide a more accurate basis for determining the dosing interval of carvedilol. PMID- 16379665 TI - Brainstem auditory-evoked response in dogs. PMID- 16379667 TI - Antidepressant-like components of Hypericum perforatum extracts: an overview of their pharmacokinetics and metabolism. AB - Extracts of Hypericum perforatum are becoming increasingly popular for the treatment of mild to moderate depression, despite the lack of consensus on their efficacy. Although the mechanism(s) of this action are still debated, several components, including the naphthodianthrones hypericin and pseudohypericin, the acylphloroglucinol hyperforin and some flavonols, are believed to play major roles in the antidepressant-like effects. Some of these also increase the expression of the P-glycoprotein transporter and others the expression of cytochrome P450 enzymes, possibly contributing to the interactions involving the extracts and conventional drugs. However, few pharmacokinetic studies of naphthodianthrones and hyperforin have appeared and none has yet evaluated the exposure to unchanged quercetin and its glycosides after intake of extracts. There are no formal pharmacokinetic studies in special populations. Bioavailability appears low, giving variable steady-state plasma concentrations, whose prediction may be complicated by non-linearity for hypericin and hyperforin. Data on tissue distribution are scarce, and it appears that hypericin and hyperforin do not reach the central nervous system in appreciable concentrations in animals. Clearance is low-intermediate, with little or no unchanged compounds excreted with urine. Although some potentially active conjugated metabolites have been identified for quercetin and its glycosides after intake of authentic compounds or flavonol-rich foods, these too have been characterised little with regard to their pharmacokinetics and central activities. Thus, further pharmacokinetic and pharmacodynamic studies of the main components and their metabolites are urgently needed to clarify the role of each constituent and provide more rational and safe regimens for people preferring "natural" drugs. PMID- 16379666 TI - Pharmacokinetics of dihydroergocristine and its major metabolite 8'-hydroxy dihydroergocristine in human plasma. AB - Dihydroergocristine (DHEC) is a semi-synthetic drug mainly used for age-related cognitive impairment. In this study, its major metabolite 8'-hydroxy dihydroergocristine (8'-OH-DHEC) was produced in incubates of a bovine liver preparation using dihydroergocristine mesylate (DHECM) as substrate. Purification was achieved by flash silica gel column and reverse phase liquid chromatographies, and identification was based on accurate molecular mass measurements, mass fragmentation spectra and NMR ((1)H/(13)C) chemical shifts. By using the substance produced in vitro, a fast, sensitive, specific and robust LC/MS/MS method for the simultaneous determination of DHEC and its major metabolite in human plasma was developed and validated. Bromocriptine was used as internal standard and limits of quantification for DHEC and 8'-OH-DHEC were 10 pg/ml and 20 pg/ml, respectively. Pharmacokinetic parameters were investigated on 12 male healthy volunteers to whom a single dose of 18 mg DHECM was administered in tablets (Iskevert). The peak of DHEC was 0.28 +/- 0.22 microg/l, the t(max) 0.46 +/- 0.26 h, the AUC(last) 0.39 +/- 0.41 microg/l.h and the terminal elimination half-life 3.50 +/- 2.27 h. The peak of 8'-OH-DHEC was 5.63 +/- 3.34 microg/l, the t(max) 1.04 +/- 0.66 h, the AUC(last) 13.36 +/- 5.82 microg/l.h and the terminal elimination half-life 3.90 +/- 1.07 h. Dosing of 18 mg DHECM was well tolerated, causing no adverse events. PMID- 16379668 TI - Role of microtubules network in CYP genes expression. AB - Superfamily of cytochrome P450 enzymes (CYPs), a distinctive enzyme system by which human body defends itself against toxic compounds, is the subject of a complex regulation process involving various mechanisms, on the levels of expression and activity. Apart from physiological factors, several patho physiological ones such as inflammation, infection, and stress affect CYP expression. The aim of this review is to summarize the current knowledge on the role of microtubules network in the regulation of drug metabolizing CYPs. Experiments on human and animal cell models revealed that microtubules disruption severely impaired basal and inducible expression of human CYP 1A1, 2B6, 2C8, 2C9, 2C19, and 3A4, and rat CYP 1A2, 2B1, 2B2, and 3A23. Inhibition of aryl hydrocarbon receptor (AhR) and glucocorticoid receptor (GR) transcriptional activity by microtubules disarray was found to be responsible for the suppressed CYP enzymes expression. However, the mechanism by which microtubules interfering agents (MIAs) inhibit GR and AhR transcriptional activities is not fully understood yet. Several lines of evidence indicate that: i) the cell cycle, G2/M phase in particular, has an influence on AhR and GR transcriptional activity, and ii) MIAs negatively modulate GR transcriptional activity via the activation of c Jun-N-terminal kinase. In conclusion, down-regulation of major CYP enzymes by microtubules disarray is intriguing from the mechanistic point of view and in relation to the cell differentiation. PMID- 16379669 TI - Improving cancer therapeutics by molecular profiling. AB - The individualized medicine aims to identify the molecular basis of the individual's response to different therapeutic treatments. Individualized medicine is very relevant for human diseases such as cancer and it has become a major task to accomplish more efficient and specific therapeutics. An individualized response to treatment could underline therapeutic success or failure and, even more, could support the rationale for good or bad prognosis. The use of up to date genomic approaches is changing the way we understand modern medicine in terms of drug efficacy, toxicity and diagnosis. Results from genetic polymorphism studies, gene expression profiling and epigenetics illustrate how pharmacogenomic testing will contribute to the goal of individualized medicine. Antineoplastic drugs are designed to block the anomalous activity of specific molecules (therapeutic targets) that regulate cellular processes such as cell cycle. Understanding the relationship between molecular changes in therapeutic targets and enhanced antitumoral response or chemotherapeutic resistance is crucial to establish the clinical relevance of genomic approaches. The goal of this review is to discuss the basic and the clinical significance of genomic research on drug targets and its impact on the early diagnosis and treatment of cancer. We will also assess how these methodologies could contribute to individualized medicine in oncology. A special focus will be put on oncogenes and tumor suppressor genes. Aspects such as drug efficacy, side effects and the diagnostic value of antineoplastic pharmacogenomic research will be also considered. PMID- 16379670 TI - A microscale in vitro physiological model of the liver: predictive screens for drug metabolism and enzyme induction. AB - In vitro models of the liver using isolated primary hepatocytes have been used as screens for measuring the metabolism, toxicity and efficacy of xenobiotics, for studying hepatocyte proliferation, and as bioartificial liver support systems. Yet, primary isolated hepatocytes rapidly lose liver specific functions when maintained under standard in vitro cell culture conditions. Many modifications to conventional culture methods have been developed to foster retention of hepatocyte function. Still, not all of the important functions -- especially the biotransformation functions of the liver -- can as yet be replicated at desired levels, prompting continued development of new culture systems. In the first part of this article, we review primary hepatocyte in vitro systems used in metabolism and enzyme induction studies. We then describe a scalable microreactor system that fosters development of 3D-perfused micro-tissue units and show that primary rat cells cultured in this system are substantially closer to native liver compared to cells cultured by other in vitro methods, as assessed by a broad spectrum of gene expression, protein expression and biochemical activity metrics. These results provide a foundation for extension of this culture model to other applications in drug discovery -- as a model to study drug-drug interactions, as a model for the assessment of acute and chronic liver toxicity arising from exposure to drugs or environmental agents; and as a disease model for the study of viral hepatitis infection and cancer metastasis. PMID- 16379671 TI - Metabolism of Delta(3)-carene by human cytochrome p450 enzymes: identification and characterization of two new metabolites. AB - The metabolism of the bicyclic monoterpene Delta(3)-carene was investigated in vitro using human liver microsomes as well as human smoker/non-smoker lung microsomes and 12 different recombinant cytochrome P450 enzymes coexpressed with human CYP-reductase in Escherichia coli cells. We detected two metabolites using GC-MS analysis. The mass fragmentation indicated for one metabolite hydroxylation in the allyl position and for the other metabolite epoxidation at the double bond. For clear identification the suggested metabolites were synthesized in a four-step reaction. Comparison of GC retention times and mass spectra lead to the identification of the metabolites as Delta(3)-carene-10-ol ((1S, 6R)-7,7 Dimethylbicyclo[4.1.0]hept-3-en-3-yl-methanol) and Delta(3)-carene-epoxide ((1S, 3S, 5R, 7R)-3,8,8-Trimethyl-4-oxa-tricyclo[5.1.0.0(3,5)]octane). Delta(3)-carene 10-ol was formed by human liver microsomes and recombinant human CYP2B6, CYP2C19 and CYP2D6. Delta(3)-Carene-epoxide was obviously catalyzed only by CYP1A2. In both cases there was a clear correlation between the metabolite formation, incubation time and enzyme concentration, respectively. Further kinetic analysis revealed that CYP2B6 exhibited the highest activity for Delta(3)-carene 10 hydroxylation. Michaelis-Menten K(m) and V(max) for oxidation of Delta(3)-carene were 0.6 mM and 28.4 nmol/min/nmol P450 using human CYP2B6. For the formation of Delta(3)-carene-epoxide 98.2 mM and 3.9 nmol/min/nmol P450 were determined as K(m) and V(max) by using human CYP1A2. To our knowledge, this is the first time that Delta(3)-carene-10-ol and Delta(3)-carene-epoxide are described as human metabolites of Delta(3)-carene. PMID- 16379673 TI - Shift work as an oxidative stressor. AB - BACKGROUND: Some medical disorders have higher prevalence in shift workers than others. This study was designed to evaluate the effect of night-shift-working on total plasma antioxidant capacity, with respect to the causative role of oxidative stress in induction of some of these disorders. METHODS: Two blood samples were taken from 44 workers with a rotational shift schedule, one after their day shift and one after their night shift. The total plasma antioxidant capacity of each worker was measured through the FRAP method. The impacts of age and weight were also assessed. RESULTS: The total plasma antioxidant capacity was measured in 44 shift-workers with a mean age of 36.57 years (SD: 10.18) and mean BMI of 26.06 (SD: 4.37) after their day and night shifts. The mean reduction of total plasma antioxidant capacity after the night shift was 105.8 micromol/L (SD: 146.39). Also, a significant correlation was shown between age and weight and total plasma antioxidant capacity. Age and weight were found to be inversely related to total plasma antioxidant capacity; as age and weight increased, the total plasma antioxidant capacity decreased. CONCLUSION: Shift work can act as an oxidative stressor and may induce many medical disorders. Aging and obesity in shift workers makes them more sensitive to this hazardous effect. PMID- 16379672 TI - Work factors and smoking cessation in nurses' aides: a prospective cohort study. AB - BACKGROUND: The prevalence of smoking in nursing personnel remains high. The aim of this study was to identify work factors that predict smoking cessation among nurses' aides. METHODS: Of 2720 randomly selected, Norwegian nurses' aides, who were smoking at least one cigarette per day when they completed a questionnaire in 1999, 2275 (83.6 %) completed a second questionnaire 15 months later. A wide spectrum of work factors were assessed at baseline. Respondents who reported smoking 0 cigarettes per day at follow-up were considered having stopped smoking. The odds ratios and 95 % confidence intervals of stopping smoking were derived from logistic regression models. RESULTS: Compared with working 1-9 hours per week, working 19-36 hours per week (odds ratio (OR) = 0.35; 95 % confidence interval (CI) = 0.13 - 0.91), and working more than 36 hours per week (i.e. more than full-time job) (OR = 0.27; CI = 0.09 - 0.78) were associated with reduced odds of smoking cessation, after adjustments for daily consumption of cigarettes at baseline, age, gender, marital status, and having preschool children. Adjusting also for chronic health problems gave similar results. CONCLUSION: There seems to be a negative association between hours of work per week and the odds of smoking cessation in nurses' aides. It is important that health institutions offer workplace-based services with documented effects on nicotine dependence, such as smoking cessation courses, so that healthcare workers who want to stop smoking, especially those with long working hours, do not have to travel to the programme or to dedicate their leisure time to it. PMID- 16379674 TI - Evidence-based cardiovascular medicine: why another journal? PMID- 16379675 TI - Focus on thrombolytic therapy. PMID- 16379676 TI - Global perspectives on cardiovascular disease. PMID- 16379677 TI - End-stage renal disease was strongly associated with increased blood pressure. PMID- 16379678 TI - Increasing blood pressure was associated with increased risk of stroke in patients with cerebrovascular disease. PMID- 16379679 TI - Low serum folate was associated with an increased risk for fatal coronary heart disease. PMID- 16379680 TI - Postmenopausal estrogen and progestin use was associated with a lower risk of cardiovascular disease. PMID- 16379681 TI - School and home interventions improved children's dietary and physical activity behaviors. PMID- 16379682 TI - Beta-carotene and vitamin A demonstrated no beneficial effect on lung cancer and death from all cause, cardiovascular cause and lung cancer. PMID- 16379683 TI - Alpha-tocopherol may reduce the risk of non-fatal myocardial infarction among patients with angiographically proven coronary atherosclerosis. PMID- 16379684 TI - Beta-carotene supplementation for 12 years did not affect the incidence of malignant neoplasms and cardiovascular disease. PMID- 16379685 TI - Detailed fetal echocardiography was sensitive and specific for the detection of congenital heart disease. PMID- 16379686 TI - Admission characteristics were weak predictors of 30-day mortality from myocardial infarction in the elderly. PMID- 16379687 TI - Routine addition of heparin to aspirin in acute myocardial infarction provides only modest benefit. PMID- 16379688 TI - Dalteparin in addition to aspirin reduces the early risk of death and new MI among patients with unstable coronary artery disease. PMID- 16379689 TI - Hirudin is modestly more effective than heparin in the treatment of acute coronary syndromes. PMID- 16379690 TI - Clopidogrel was more efficacious than aspirin in preventing major cardiovascular events. PMID- 16379691 TI - Continuous intravenous epoprostenol improved survival, clinical status and hemodynamics in patients with severe primary pulmonary hypertension. PMID- 16379692 TI - Right heart catheterization was associated with increased mortality and hospital costs. PMID- 16379693 TI - Five-year survival rates following PTCA and CABG were not significantly different. PMID- 16379694 TI - Endarterectomy is not indicated for moderate symptomatic carotid stenosis. PMID- 16379695 TI - Lower than optimal use of thrombolytic therapy, particularly with increasing age and female sex. PMID- 16379696 TI - Use of statins in primary or secondary prevention of coronary heart disease is cost-effective in some but not all. PMID- 16379697 TI - Impact of carvedilol on mortality and cardiovascular morbidity in patients with chronic heart failure. PMID- 16379698 TI - From journal to bedside: application of clinical trial results to individual patients. PMID- 16379699 TI - Focus on congestive heart failure. PMID- 16379700 TI - Lipid lowering in patients with coronary artery disease: low density lipoprotein cholesterol and beyond. AB - Now that the importance of LDL-C and its reduction are well established in the prevention of atherosclerotic vascular complications, we are moving to a new era in which physicians must pay more attention to factors beyond LDL-C lowering. More emphasis should be put on TRL and remnant lipoproteins as well as other contributors to the cardiovascular risk burden, such as thrombotic risk factors and impaired fibrinolysis. This should be carried out within the standard framework of a global approach to risk factor management in CAD patients. PMID- 16379701 TI - Hypertension was the most common cause of congestive heart failure. PMID- 16379702 TI - Surgical correction of mitral regurgitation due to flail leaflet was associated with improved long-term survival. PMID- 16379704 TI - Elevated cardiac troponin T levels were associated with increased 30-day mortality in patients presenting with acute myocardial ischemia. PMID- 16379703 TI - Ankle brachial index is a predictor of 5-year cardiovascular events. PMID- 16379705 TI - Elevated cardiac troponin I levels on admission were associated with increased 42 day mortality in patients with acute ischemic syndromes. PMID- 16379706 TI - Markers of left ventricular dysfunction were better predictors of a poor prognosis than markers of ischemia. PMID- 16379707 TI - Transdermal nicotine was safe as an aid to smoking cessation in participants with cardiac disease. PMID- 16379708 TI - Moderate-dose anticoagulation was as effective in preventing thromboembolic events as conventional-dose anticoagulation in patients with mechanical heart valves. PMID- 16379709 TI - Digoxin did not reduce mortality but reduced the rate of hospitalization due to worsening heart failure among patients with heart failure. PMID- 16379710 TI - No effect of amlodipine on mortality or cardiovascular morbidity in patients with severe chronic heart failure. PMID- 16379711 TI - Prophylactic implanted defibrillator may improve survival in patients at high risk for ventricular tachyarrhythmias. PMID- 16379712 TI - Stent implantation reduced the rate of restenosis following successful recanalization of chronic coronary occlusions. PMID- 16379713 TI - Antiplatelet therapy reduced the incidence of cardiac, hemorrhagic and vascular complications following coronary artery stent placement. PMID- 16379714 TI - Aggressive as opposed to moderate low-density lipoprotein cholesterol lowering strategies reduced the progression of atherosclerosis in grafts and reduced further revascularization. PMID- 16379715 TI - Moderate intensity adjusted-dose warfarin was more effective than low-intensity fixed dose warfarin and aspirin in preventing strokes in atrial fibrillation. PMID- 16379716 TI - Streptokinase in the early treatment of acute ischemic stroke was associated with increased mortality. PMID- 16379717 TI - Selection of patients for angiography following myocardial infarction was not based on risk but on young age and availability of angiographic facilities. PMID- 16379718 TI - Inverse correlation between increasing local rates of coronary artery bypass graft and the average anticipated survival benefit. PMID- 16379719 TI - PTCA had little cost advantage over CABG in patients with multi-vessel disease. PMID- 16379721 TI - From journal to bedside: quantifying the benefits of treatment. PMID- 16379720 TI - Atenolol before and after non-cardiac surgery reduced mortality and cardiovascular morbidity in patients at risk of acute coronary events. PMID- 16379722 TI - Management of the patient in atrial fibrillation. PMID- 16379723 TI - An approach to treating hypertension. PMID- 16379724 TI - Higher socio-economic status was associated with fewer coronary events and lower case-fatality rates. PMID- 16379725 TI - Reduction in CHD mortality was explained by both secondary risk factor reduction and improved treatment of CHD. PMID- 16379726 TI - High fish consumption was associated with a lower 30-year risk of death from coronary heart disease. PMID- 16379728 TI - Obesity, cigarette smoking and hypertension were associated with increased risk of pulmonary embolism in women. PMID- 16379727 TI - Higher plasma C-reactive protein was associated with increased risk of first thrombotic event. PMID- 16379729 TI - Continued smoking following successful percutaneous coronary revascularization increased the risk of all-cause mortality and Q-wave MI. PMID- 16379730 TI - Fruit-and-vegetable rich diet low in fat significantly reduced blood pressure. PMID- 16379731 TI - Realistic dietary changes in type, but not amount of, dietary fat reduced LDL and total blood cholesterol. PMID- 16379732 TI - Diuretics and beta-blockers are safe and effective first-line anti-hypertensive therapies. PMID- 16379733 TI - Low-dose chlorthalidone was beneficial in the treatment of isolated systolic hypertension in non-insulin-dependent diabetic and non-diabetic patients. PMID- 16379734 TI - Dual-chamber pacing produced modest symptomatic and hemodynamic improvements in patients with hypertrophic cardiomyopathy. PMID- 16379736 TI - Amiodarone reduced the risk of resuscitated ventricular fibrillation or arrhythmic death in survivors of MI with frequent or repetitive ventricular premature depolarizations. PMID- 16379735 TI - Active compression-decompression CPR improved short-term survival. PMID- 16379737 TI - Amiodarone reduced arrhythmic but not all-cause mortality among MI survivors with depressed LV function. PMID- 16379738 TI - Transmyocardial laser revascularization may relieve angina and improve myocardial perfusion among patients with angina refractory to medical treatment. PMID- 16379739 TI - Peri-operative supplemental warming was associated with fewer morbid cardiac events than routine thermal care. PMID- 16379740 TI - Adverse cerebral events were common and were associated with longer hospital stays. PMID- 16379741 TI - Verapamil improved walking distance in patients with claudication. PMID- 16379742 TI - Poor blood pressure control among treated hypertensives was not attributable to a 'white-coat' effect. PMID- 16379744 TI - Losartan was associated with a similar rate of renal dysfunction and worsening heart failure as captopril with fewer adverse effects. PMID- 16379743 TI - Population-wide educational strategies for serum cholesterol reduction are cost effective. PMID- 16379745 TI - From journal to bedside: obtaining medically useful answers from a meta-analysis. PMID- 16379746 TI - Dysglycemia, not just diabetes, is a continuous risk factor for cardiovascular disease. AB - Dysglycemia is a continuous cardiovascular risk factor. Whether or not interventions that lower glucose levels will also lower cardiovascular risk remains to be determined. A number of ongoing studies may provide answers to this question. Recognition that these non-diabetic glucose levels confer an increased cardiovascular risk may suggest new ways for preventing CVD. PMID- 16379747 TI - Focus on unstable angina. PMID- 16379748 TI - Passive smoking was associated with an increased risk of CHD. PMID- 16379749 TI - U-shaped relationship between BMI and risk of cardiovascular morbidity and mortality. PMID- 16379750 TI - Obesity and weight gain were associated with increased risk of ischemic, but not hemorrhagic stroke. PMID- 16379751 TI - LV internal dimensions predicted an increased risk of CHF in patients free of MI. PMID- 16379752 TI - Aortic jet velocity and baseline functional score predicted aortic valve replacement and death. PMID- 16379753 TI - Enoxaparin reduced the rate of the composite end-point of recurrent angina, MI and death in unstable angina. PMID- 16379754 TI - Subcutaneous dalteparin was similar to intravenous heparin in the early treatment of unstable angina. PMID- 16379755 TI - Abciximab with low-dose heparin prevented complications following percutaneous coronary revascularization without increased bleeding risk. PMID- 16379756 TI - Abciximab reduced short-term rates of death, MI and repeat revascularization in patients with refractory unstable angina undergoing PTCA. PMID- 16379757 TI - Eptifibatide did not significantly reduce the rate of adverse events in patients undergoing percutaneous coronary revascularization. PMID- 16379758 TI - Beta-blockers appear to reduce mortality in patients with heart failure. PMID- 16379759 TI - Oral theophylline may play a role in the treatment of sinus node disease. PMID- 16379760 TI - Rampiril reduced long-term mortality in patients with clinical heart failure following acute MI. PMID- 16379761 TI - Primary angioplasty reduced rate of death, reinfarction or disabling stroke. PMID- 16379762 TI - Catheter-based radiotherapy prevented coronary restenosis after stenting in patients with previous restenosis. PMID- 16379764 TI - Heparin and aspirin did not prevent early deaths following ischemic stroke. PMID- 16379763 TI - Probucol reduced the rates of restenosis following scheduled PTCA. PMID- 16379765 TI - Aspirin modestly decreased rate of death or recurrent ischemic stroke. PMID- 16379766 TI - High physician and hospital angioplasty volumes resulted in better cardiac outcomes. PMID- 16379767 TI - High-volume interventionalists had fewer major complications than low-volume interventionalists. PMID- 16379768 TI - Oral potassium reduced systolic and diastolic BP in hypertensive and normotensive participants. PMID- 16379769 TI - Drug safety: the different perspectives. PMID- 16379770 TI - Focus on the management of a sudden death survivor. PMID- 16379771 TI - Treatment for high blood pressure: evidence and practice. PMID- 16379772 TI - Smoking, high SBP and cholesterol levels were associated with increased risk of carotid artery stenosis. PMID- 16379773 TI - Cynical hostility was associated with increased risk of mortality and MI. PMID- 16379774 TI - Calcium supplementation during pregnancy was associated with lower SBP in the children. PMID- 16379775 TI - On-site cardiac catheterization facilities were associated with more cardiac procedures but no difference in mortality. PMID- 16379776 TI - Medium-dose hirudin was superior to heparin in preventing ischemic events in patients with acute myocardial ischemia. PMID- 16379777 TI - Ramipril following acute MI attenuated LV remodeling. PMID- 16379778 TI - PTCA in comparison to medical therapy was associated with a significant excess of death and non-fatal MI among patients with angina. PMID- 16379779 TI - Deferred invasive treatment was associated with fewer MIs in patients with inducible ischemia post-AMI. PMID- 16379780 TI - Reteplase was not better than alteplase in reducing 30-day mortality among patients with acute MI. PMID- 16379781 TI - Double-bolus alteplase was not equivalent to accelerated infusion in patients with acute MI. PMID- 16379782 TI - Felodipine in addition to enalapril did not improve clinical status, functional capacity or ventricular function in patients with CHF. PMID- 16379783 TI - Smoking cessation interventions improved the 12-month smoking cessation rate among patients undergoing non-cardiac surgery. PMID- 16379784 TI - Reductase inhibitor monotherapy for cholesterol reduction lowered the incidence of stroke. PMID- 16379785 TI - Administration of captopril following MI reduced the incidence of ischemia related events. PMID- 16379786 TI - Anti-hypertensive therapy reduced the incidence of stroke in older patients with isolated systolic hypertension. PMID- 16379787 TI - Ramipril slowed the decline in renal function among patients with nephropathy and proteinuria not due to insulin-dependent diabetes mellitus. PMID- 16379788 TI - Lisinopril inhibited the progression of renal albumin excretion in patients with insulin-dependent diabetes mellitus. PMID- 16379789 TI - Thrombolytic therapy increases the frequency of death and intra-cranial hemorrhage in survivors of ischemic stroke. PMID- 16379790 TI - Epoprostenol increased mortality in patients with severe CHF. PMID- 16379791 TI - Psycho-social nursing interventions following acute MI were not effective. PMID- 16379792 TI - Hyperventilation was a highly-specific test for the detection of coronary artery spasm but with only modest sensitivity. PMID- 16379793 TI - Asymptomatic LV systolic dysfunction was more than twice as prevalent as symptomatic LV systolic dysfunction. PMID- 16379794 TI - Elevated plasma homocysteine was associated with increased mortality. PMID- 16379795 TI - Elevated plasma homocysteine was associated with increased risk of vascular disease. PMID- 16379797 TI - 2-year survival with atenolol administration. PMID- 16379798 TI - Understanding the fundamentals of quality of life measurement. PMID- 16379799 TI - Selection of patients for coronary revascularization by PTCA. PMID- 16379800 TI - How the cardiovascular burden of illness is changing in eastern Europe. PMID- 16379801 TI - Cigarette smoking was associated with an increased risk of mortality in China. PMID- 16379802 TI - Smoking is a major cause of death in China. PMID- 16379803 TI - Environmental tobacco smoke exposure was associated with an increased risk of ischemic heart disease. PMID- 16379804 TI - Children of parents with early CAD were more likely to have an adverse risk profile by early adulthood. PMID- 16379805 TI - Substitution of saturated and trans unsaturated fats with mono- and poly unsaturated fats reduced the risk of CHD. PMID- 16379806 TI - Ticlopidine reduced the incidence of late occlusion of below-the-knee saphenous vein grafts. PMID- 16379807 TI - Prophylactic oral amiodarone following cardiac surgery reduced the rate of atrial fibrillation and lowered hospitalization costs. PMID- 16379808 TI - Long-term single-chamber atrial pacing for sick-sinus syndrome was associated with fewer deaths and adverse events. PMID- 16379809 TI - Prophylactic use of implantable cardiac defibrillators was not associated with improved survival in patients undergoing elective CABGS. PMID- 16379810 TI - ICD improved survival in patients resuscitated from life-threatening ventricular arrhythmias. PMID- 16379811 TI - Amiodarone reduced total mortality and arrhythmic/sudden death in patients with recent MI or with CHF. PMID- 16379812 TI - Bupropion was efficacious and safe as an aid to smoking cessation. PMID- 16379813 TI - The use of calcium antagonists and ACE inhibitors increased between 1992 and 1995. PMID- 16379814 TI - Plasma BNP concentration predicted the presence of heart failure. PMID- 16379815 TI - Brain natriuretic peptide concentration was effective for screening for LV systolic dysfunction. PMID- 16379816 TI - Dietary changes and exercise significantly retarded atherosclerosis and improved work capacity in patients with CAD. PMID- 16379817 TI - Sexual maturation, BMI, dietary intervention and dietary cholesterol reduced LDL cholesterol in children. PMID- 16379818 TI - Moderately reduced dietary fat intake was associated with reduced LDL cholesterol in hypercholesterolemic and combined hyperlipidemic men. PMID- 16379819 TI - Interpreting results of trials using surrogate outcome measures. PMID- 16379820 TI - Management of pregnant women with prosthetic heart valves. PMID- 16379821 TI - Role of psycho-social factors in CVD. PMID- 16379822 TI - Elevated concentrations of soluble intercellular adhesion molecule-1 were associated with increased risk of MI. PMID- 16379823 TI - Increased 1-hour postload plasma glucose was associated with increased risk of fatal CHD. PMID- 16379824 TI - There was a weak association between insulin levels and risk of CVD. PMID- 16379825 TI - Factors enhancing the effect of warfarin anti-coagulation. PMID- 16379826 TI - The risk of congenital heart disease recurrence in the offspring of adults with major heart defects. PMID- 16379827 TI - Multiple cardiovascular risk factors in young people were associated with marked increase in coronary atherosclerosis. PMID- 16379828 TI - Ischemic heart disease mortality declined sharply in Poland: an ecological study. PMID- 16379829 TI - Thallium perfusion defects and impaired exercise capacity predict death and non fatal MI in symptom-free patients with previous CABG. PMID- 16379830 TI - The incidence of infective endocarditis following surgical repair of a major congenital defect varied by type of heart defect. PMID- 16379831 TI - Balloon and closed mitral commissurotomy were associated with superior early hemodynamic and late functional and echocardiographic results. PMID- 16379832 TI - Nisoldipine was associated with increased risk of MI among hypertensive diabetic patients when compared with enalapril. PMID- 16379833 TI - Directional coronary atherectomy reduced the rate of late restenosis compared to PTCA in patients with stable or unstable angina. PMID- 16379834 TI - Surgery was beneficial for severe carotid stenosis in patients with symptomatic carotid ischemia. PMID- 16379835 TI - Dual-chamber pacing was more effective in patients with sinus-node dysfunction. PMID- 16379836 TI - The combination of warfarin and aspirin reduced the rate of IHD events more effectively than either agent alone. PMID- 16379837 TI - Primary coronary angioplasty is superior to thrombolytic therapy for acute MI. PMID- 16379838 TI - Urokinase reduced the need for open surgical procedures in acute ischemia of the leg. PMID- 16379839 TI - Proficiency in cardiac auscultation was poor in internal medicine and family practice residents. PMID- 16379840 TI - Physicians identified smoking status in two-thirds of office visits and provided cessation counseling to only a minority of smokers. PMID- 16379841 TI - Isosorbide dinitrate and furosemide in the treatment of patients with severe pulmonary edema. PMID- 16379842 TI - Assessing the incremental value of diagnostic tests. PMID- 16379843 TI - Hypertrophic cardiomyopathy. PMID- 16379844 TI - Epidemiology and prevention of rheumatic fever and current issues in the prevention of rheumatic heart disease. PMID- 16379845 TI - Targeting three subgroups of hypertensives can reduce incidence of intracerebral hemorrhage. PMID- 16379846 TI - Combination of diet and exercise effective in reducing LDL-cholesterol. PMID- 16379847 TI - Estrogen-progestin therapy was not effective in secondary prevention of CHD in post-menopausal women. PMID- 16379848 TI - Ischemic ST-segment response is a significant predictor of coronary events in asymptomatic individuals. PMID- 16379849 TI - Baroreflex sensitivity and heart rate variability were associated with total cardiac mortality following MI. PMID- 16379850 TI - Poor long-term survival after acute MI in patients receiving dialysis. PMID- 16379851 TI - Lovastatin is effective in primary prevention of acute coronary events. PMID- 16379852 TI - Beta-blockers were not as efficacious as diuretics when used as first-line therapy in elderly patients with uncomplicated hypertension. PMID- 16379853 TI - Garlic tablets are ineffective in hypercholesterolemia. PMID- 16379854 TI - Tirofiban combined with heparin and aspirin prevented ischemic events in patients with unstable angina and non-Q-wave MI. PMID- 16379856 TI - Efficacy of eptifibatide in patients with acute coronary syndromes. PMID- 16379855 TI - Tirofiban plus aspirin reduced ischemic events in patients with unstable angina. PMID- 16379857 TI - Use of abciximab is beneficial in stenting and balloon angioplasty. PMID- 16379858 TI - Stenting is superior to balloon angioplasty alone in preventing repeat PTCA. PMID- 16379859 TI - Beta-blockers are under-utilized in elderly patients after MI. PMID- 16379860 TI - Use of beta-blockers was associated with reduced mortality in high-risk patients. PMID- 16379861 TI - Home-based intervention strategy was superior to usual care in discharged CHF patients. PMID- 16379862 TI - Process of care is affected by the type of health insurance. PMID- 16379863 TI - Invasive treatment is not superior to conservative treatment in patients with non Q-wave MI. PMID- 16379865 TI - Role of local medical opinion leaders in adoption of AMI guidelines. PMID- 16379864 TI - Variation in use of invasive procedures in acute ischemic syndromes. PMID- 16379866 TI - Plasma LDL concentrations achieved by pravastatin reduced coronary event rate. PMID- 16379867 TI - Therapeutic objectives. PMID- 16379868 TI - What are my chances? PMID- 16379869 TI - Applying evidence-based cardiology to the individual patient. PMID- 16379871 TI - Why physicians should read reports of an economic analysis. PMID- 16379870 TI - Unique contributions of observational evidence. PMID- 16379872 TI - Subgroups time to change the question? PMID- 16379873 TI - The care gap: improving delivery of patient care. PMID- 16379875 TI - Renal function: an emerging risk factor for cardiovascular disease? PMID- 16379876 TI - Overcoming the evidence-to-application gap in clinical practice. PMID- 16379877 TI - Pharmacogenetics in Evidence-based practice--what to watch for. PMID- 16379878 TI - Managing acute and recent-onset atrial fibrillation. PMID- 16379880 TI - Finding evidence when you need it. PMID- 16379881 TI - Meta analysis finds strong evidence that aspirin reduces the risk of first myocardial infarction. PMID- 16379882 TI - Trial suggests diuretic based antihypertensive treatment is more effective than newer alpha blocker based treatments for preventing cardiovascular disease. PMID- 16379883 TI - Trial suggests that combining eplerenone and enalapril reduces left ventricular hypertrophy in hypertension more effectively than either treatment alone. PMID- 16379885 TI - Meta-analysis suggests regression of left ventricular hypertrophy during antihypertensive treatment is linked to reduced risk of cardiovascular disease. PMID- 16379886 TI - Trial suggests once daily lisinopril is safe and effective for hypertensive children. PMID- 16379887 TI - Trial suggests nurse counselling may reduce antihypertensive medication, but not alter blood pressure in high risk people with hypertension. PMID- 16379888 TI - Cohort study finds that impatience and hostility in young adults are linked with long-term risk of hypertension. PMID- 16379889 TI - Trial suggests initial angioplasty improves angina symptoms and exercise tolerance, but has no survival benefit over continued medical treatment. PMID- 16379891 TI - Trial finds sustained release propafenone increases time to recurrent atrial fibrillation compared with placebo. PMID- 16379892 TI - Trial suggests that prolonged antithrombotic pretreatment does not improve outcomes over early intervention in unstable coronary syndromes. PMID- 16379894 TI - Trial finds sirolimus eluting stents reduce restenosis in people with complex coronary lesions. PMID- 16379896 TI - Trial finds that exercise testing the day after coronary stenting is safe. PMID- 16379895 TI - Trial finds sirolimus eluting stents reduce restenosis in people with long atherosclerotic lesions in small coronary arteries. PMID- 16379897 TI - Trial suggests that perindopril reduces major cardiovascular events among people with stable coronary artery disease. PMID- 16379898 TI - Trial suggests long term trapidil reduces cardiovascular events and improves prognosis in coronary artery disease. PMID- 16379899 TI - Meta analysis suggests that intravenous GP IIb/IIIa antagonists improve short term survival following percutaneous coronary intervention. PMID- 16379900 TI - Meta analysis suggests myocardial viability affects long term survival following revascularisation in left ventricular dysfunction and coronary artery disease. PMID- 16379901 TI - Trial suggests transferring people with acute myocardial infarction for angioplasty is more effective than thrombolysis in a non interventional centre. PMID- 16379902 TI - Meta analysis suggests transfer to an angioplasty centre is superior to immediate thrombolysis in acute myocardial infarction. PMID- 16379903 TI - Review finds that telemedicine may reduce hospital readmissions in heart failure, but further evidence is needed. PMID- 16379904 TI - Trial finds candesartan reduces cardiovascular deaths and hospital admissions in people with heart failure, but may not affect all cause mortality. PMID- 16379906 TI - Meta analysis suggests that non lipid serum marker effects can not be used to differentiate between statins. PMID- 16379909 TI - Does evidence-based medicine do more harm than good for the elderly? PMID- 16379907 TI - Cohort study finds that combined elevated C-reactive protein and erythrocyte sedimentation rate predicts 6 month outcome in people with chest pain, but non diagnostic electrocardiogram and troponin T. PMID- 16379910 TI - Trial finds that omapatrilat is more effective than enalapril for reducing hypertension. PMID- 16379912 TI - Cohort study suggests that intensive patient education helps modify lifestyle risk factors in people with hypertension. PMID- 16379911 TI - Trial finds 6-monthly follow-up is as effective as 3-monthly follow-up for controlling hypertension in primary care. PMID- 16379913 TI - Trial finds verapamil and atenolol-based antihypertensive regimens are similarly effective in coronary artery disease. PMID- 16379914 TI - Trial finds ranolazine increases exercise tolerance in people with severe angina receiving atenolol, amlodipine or diltiazem. PMID- 16379915 TI - Review finds evidence to support pharmacological treatment in many areas of atrial fibrillation, but no strong evidence for one drug over another. PMID- 16379917 TI - Trial finds intravenous sotalol may control heart rate rapidly in atrial fibrillation where long-term use is intended. PMID- 16379916 TI - Trial finds amiodarone treatment maintains sinus rhythm safely in persistent atrial fibrillation. PMID- 16379918 TI - Trial finds adding abciximab to stent reperfusion improves survival in acute myocardial infarction. PMID- 16379919 TI - Trial finds paclitaxel-eluting stents reduce restenosis over bare metal stents in coronary artery disease. PMID- 16379921 TI - Trial finds that direct stenting does not reduce restenosis compared to conventional stenting in unselected lesions. PMID- 16379923 TI - Review finds insufficient evidence that arterial puncture closing devices are superior to standard manual compression. PMID- 16379922 TI - Trial finds that abciximab does not improve 30-day outcomes following elective PCI with clopidogrel pre-treatment. PMID- 16379924 TI - Trial finds off-pump coronary artery bypass surgery is as safe as on-pump surgery for people requiring multivessel grafting, but may have lower graft patency at 3 months. PMID- 16379926 TI - Meta-analysis finds that exercise training improves survival in people with heart failure. PMID- 16379925 TI - Trial finds that simvastatin plus niacin is safe in people with coronary artery disease and low HDL cholesterol. PMID- 16379927 TI - Review finds inadequate evidence of the cost effectiveness and efficacy of open access echocardiography in primary care. PMID- 16379928 TI - Trial finds that chest pain observation units reduce hospital admissions without increasing costs compared to usual care. PMID- 16379929 TI - Trial suggests vasopressin is as effective as epinephrine for out of hospital cardiopulmonary resuscitation and may be superior for people with asystole. PMID- 16379930 TI - Review finds that high intensity vitamin K antagonist therapy reduces thrombosis and bleeding more than low intensity therapy for people with mechanical heart valves. PMID- 16379931 TI - Criticisms of evidence-based medicine. PMID- 16379932 TI - Review finds that simplifying antihypertensive regimens may improve treatment adherence. PMID- 16379933 TI - Trial finds adjusting antihypertensive drugs based on home measurement may not improve blood pressure control. PMID- 16379934 TI - Trial finds that enoxaparin may be the preferred drug for initiating anticoagulation in atrial fibrillation. PMID- 16379935 TI - Trial finds direct stenting has similar vessel trauma and angiographic outcomes to stenting following predilation. PMID- 16379936 TI - Review finds primary stenting lowers the risk of reinfarction and revascularisation, but does not improve survival compared to balloon angioplasty. PMID- 16379937 TI - Trial finds cutting balloon angioplasty has procedural benefits over conventional angioplasty, but few clinical advantages. PMID- 16379938 TI - Review finds that ablative therapies do not improve clinical or angiographic outcomes compared to balloon dilation. PMID- 16379939 TI - Trial finds no evidence that directional coronary atherectomy prior to stenting has any benefit over stenting alone. PMID- 16379940 TI - Trial finds people with acute myocardial infarction ineligible for thrombolysis may benefit from angioplasty, with or without stenting. PMID- 16379941 TI - Trial finds clopidogrel is similarly tolerable to ticlopidine after stenting. PMID- 16379942 TI - Trial finds stenting for multivessel disease is less costly than bypass grafting, but more likely to require repeat revascularisation. PMID- 16379943 TI - Review finds combining angiotensin receptor blockers with ACE inhibitors in heart failure may only benefit people with contraindications to beta blockers. PMID- 16379944 TI - Digitalis may reduce hospitalisation and improve clinical status in heart failure, but has no effect on mortality. PMID- 16379945 TI - Trial finds using recent symptoms in computer-generated care suggestions does not improve outcomes in heart failure. PMID- 16379946 TI - Trial finds nurses can safely and effectively prepare patients for diagnostic cardiac catheterisation. PMID- 16379947 TI - Review finds discharge planning and support reduces hospital readmission for older people with heart failure. PMID- 16379948 TI - Review finds limited evidence that 'stages of change' interventions modify behaviour in primary care. PMID- 16379949 TI - Trial finds regular exercise may improve survival and lower costs compared to percutaneous coronary interventions in stable coronary artery disease. PMID- 16379950 TI - Review finds further work is required to define and assess the incidence of aspirin resistance. PMID- 16379951 TI - Trial finds intensive lipid-lowering reduces coronary atherosclerosis progression more than moderate regimen. PMID- 16379952 TI - Trial finds low dose ramipril does not reduce cardiovascular or renal events in people with type 2 diabetes and albuminuria. PMID- 16379953 TI - Review finds early invasive therapy may improve outcomes in unstable angina and non-ST-segment elevation myocardial infarction. PMID- 16379954 TI - Evidence-based Cardiovascular Medicine--an essential resource for busy practitioners. PMID- 16379956 TI - Trial finds exercise does not slow atherosclerosis in middle-aged men receiving statins. PMID- 16379955 TI - Quantifying the gap between proof and practice. PMID- 16379957 TI - Cost analysis suggests following evidence-based guidelines could reduce prescription costs in hypertension. PMID- 16379958 TI - Trial finds adding amlodipine to benazepril reduces diastolic blood pressure in non-responders to benazepril monotherapy. PMID- 16379959 TI - Trial finds valsartan and amlodipine equally effective for preventing cardiac disease among people with hypertension at high risk. PMID- 16379960 TI - Meta-analysis finds women with dyslipidemia respond as well or better than men to extended-release niacin. PMID- 16379962 TI - Trial finds administering carvedilol in hospital improves treatment adherence for people with heart failure. PMID- 16379961 TI - Trial finds no evidence that aspirin and warfarin improve outcomes in people with heart failure. PMID- 16379963 TI - Trial finds tolvaptan may help manage congestion in people with heart failure, but did not reduce death or hospitalisation. PMID- 16379964 TI - Review suggests benefits of beta-blockers may outweigh adverse effects in people with heart failure and systolic dysfunction. PMID- 16379965 TI - Meta-analysis finds exercise training is safe in people with heart failure and left ventricular dysfunction. PMID- 16379966 TI - Trial finds new anti-arrhythmic, tedisamil, is superior to placebo for converting recent onset atrial fibrillation or atrial flutter. PMID- 16379968 TI - Trial finds routine intravenous glucose-insulin-potassium does not improve myocardial salvage in people with acute myocardial infarction. PMID- 16379967 TI - Trial finds administering pravastatin early after symptom onset does not improve outcomes in people with acute coronary syndromes. PMID- 16379969 TI - Trial finds carbon-coated stents do not reduce restenosis more than bare metal stents. PMID- 16379970 TI - Trial finds rescue angioplasty does not improve survival in people with STEMI in whom fibrinolysis is unsuccessful. PMID- 16379971 TI - Trial finds high-dose bolus tirofiban is safe and effective for preventing complications in high-risk percutaneous coronary intervention. PMID- 16379972 TI - Meta-analysis finds exercise-based rehabilitation improves survival in people with coronary heart disease. PMID- 16379973 TI - Review finds coronary artery calcium score is an independent predictor of coronary events. PMID- 16379974 TI - Trial finds low carb diets reduce weight more than low fat diets in the short term, but have more minor adverse effects. PMID- 16379977 TI - Cell transplantation for heart disease: the clinical perspective. PMID- 16379978 TI - Taking diltiazem ER in the evening reduces morning blood pressure and heart rate more than ramipril. Commentary. PMID- 16379980 TI - Some antihypertensives are more effective than others for black people . Commentary. PMID- 16379979 TI - Atenolol is less effective than other antihypertensive drugs. Commentary 1. PMID- 16379981 TI - Calcium channel blockers reduce risk of stroke in people with hypertension. Commentary. PMID- 16379982 TI - Nifedipine is safe for people with angina. Commentary. PMID- 16379983 TI - Amiodarone and sotalol appear similarly safe and effective for reducing atrial fibrillation following open heart surgery. Commentary. PMID- 16379984 TI - Antiplatelet therapy plus moderate intensity anticoagulation is safe and effective in people with atrial fibrillation. Commentary. PMID- 16379985 TI - Angiotensin receptor blockers and ACE inhibitors are similarly effective in chronic heart failure and in high-risk acute myocardial infarction. Commentary. PMID- 16379986 TI - Actinomycin D-eluting stents do not reduce the risk of restenosis. Commentary. PMID- 16379987 TI - QuaDDS stents have an unacceptable safety profile. Commentary. PMID- 16379988 TI - PC-coated stents and abciximab do not reduce restenosis more than angioplasty in small coronary vessels. Commentary. PMID- 16379989 TI - Culprit lesion revascularisation initially costs less and is more successful than complete revascularisation in multivessel disease, but results equalise over time. Commentary. PMID- 16379990 TI - Early post-thrombolysis catheterisation may be preferable to a conservative strategy for people with ST-elevation myocardial infarction. Commentary 1. PMID- 16379991 TI - Early post-thrombolysis catheterisation may be preferable to a conservative strategy for people with ST-elevation myocardial infarction. Commentary. PMID- 16379992 TI - Coronary stenting improves myocardial salvage more than angioplasty when thrombolysis fails in acute myocardial infarction. Commentary. PMID- 16379993 TI - PCI with bivalirudin and provisional GP IIb/IIIa inhibition is more cost effective than heparin plus routine GP IIb/IIIa inhibition. Commentary. PMID- 16379994 TI - Posting treatment recommendations to primary care physicians does not improve care for people with myocardial infarction or heart failure. Commentary. PMID- 16379995 TI - Aggressive statin therapy in disease management clinics reduces cardiovascular morbidity for people with coronary heart disease. Commentary. PMID- 16379996 TI - Pravastatin may prevent disease progression in normocholesterolemic people with coronary artery disease. Commentary. PMID- 16379997 TI - Anti-tachycardia pacing for rapid ventricular tachycardia is as effective as shocks in people with implantable cardioverter defibrillators. Commentary. PMID- 16379998 TI - Transforming chronic care: a systematic review of the evidence. PMID- 16379999 TI - Fibre supplementation may help to reduce blood pressure. Commentary. PMID- 16380000 TI - Higher starting doses of atorvastatin may reduce LDL-cholesterol levels more than lower doses. Commentary. PMID- 16380001 TI - Simvastatin may have a slight effect on cognitive functioning in people with hypercholesterolemia. Commentary. PMID- 16380002 TI - Regimen to increase HDL cholesterol levels may improve cholesterol profiles. Commentary. PMID- 16380003 TI - Regimen to increase HDL cholesterol levels may improve cholesterol profiles. Commentary 2. PMID- 16380004 TI - Single-lead, shock-only ICD therapy reduces sudden death in people with congestive heart failure. Commentary. PMID- 16380005 TI - Valsartan reduces morbidity in people with heart failure receiving a low dose of ACE inhibitor and no beta-blocker. Commentary. PMID- 16380006 TI - ACE inhibitors improve exercise duration slightly in people with systolic heart failure. Commentary. PMID- 16380007 TI - Alternative energies may be as effective as classic Cox-Maze cut and sew techniques in the surgical treatment of atrial fibrillation. Commentary. PMID- 16380008 TI - Perioperative amiodarone reduced the incidence of atrial fibrillation after CABG but high thoracic epidural anesthesia did not. Commentary. PMID- 16380009 TI - Paclitaxel and sirolimus-eluting stents may have similar outcomes in routine practice. Commentary. PMID- 16380010 TI - Abciximab improves clinical outcomes in people with acute myocardial infarction receiving stents. Commentary. PMID- 16380011 TI - Routine stenting, but not abciximab, improves outcomes for people with diabetes and acute myocardial infarction receiving primary PCI. Commentary. PMID- 16380012 TI - Telephone calls from nurses can improve symptoms, anxiety, and knowledge among ICD recipients. Commentary. PMID- 16380013 TI - Multifaceted outpatient support can improve outcomes for people with heart failure. Commentary. PMID- 16380014 TI - There is insufficient evidence about the efficacy of commercial weight loss programmes. Commentary. PMID- 16380015 TI - Statins slow the progression of atherosclerosis and improve vascular function. Commentary. PMID- 16380016 TI - Aspirin and lipid-lowering agents reduce cardiovascular events following CABG, but there is little evidence about other therapies. Commentary. PMID- 16380017 TI - Translating evidence into practice for providers and patients. PMID- 16380018 TI - There may be gender differences in acute myocardial infarction triggers. Commentary. PMID- 16380019 TI - Antiplatelet therapy can be recommended for secondary but not primary prevention in people with hypertension. Commentary 1. PMID- 16380020 TI - Antiplatelet therapy can be recommended for secondary but not primary prevention in people with hypertension. Commentary 2. PMID- 16380021 TI - Adding magnesium sulfate to usual care may improve rate and rhythm control for people with rapid atrial fibrillation. Commentary. PMID- 16380022 TI - Invasive strategy should be considered for all people with unstable angina or non ST segment elevation myocardial infarction. Commentary. PMID- 16380023 TI - Adding carvedilol to ACE inhibitors may reduce arrhythmia and ventricular tachycardia following acute myocardial infarction. Commentary. PMID- 16380024 TI - Enoxaparin appears as effective as unfractionated heparin for people with high risk non-ST elevation acute coronary syndromes receiving rapid invasive therapy. Commentary. PMID- 16380025 TI - Drug-eluting stents do not increase the risk of stent thrombosis, but longer stents may be associated with greater incidence of thrombosis. Commentary. PMID- 16380026 TI - Paclitaxel-eluting stents have no greater risk of stent thrombosis in the first year than bare metal stents. Commentary. PMID- 16380027 TI - Trapidil does not reduce major cardiac events following balloon angioplasty. Commentary. PMID- 16380028 TI - B-type natriuretic peptide is a strong prognostic indicator for people with heart failure. Commentary. PMID- 16380029 TI - Ambulatory blood pressure monitoring may be a prognostic marker in people with heart failure. Commentary 1. PMID- 16380030 TI - Ambulatory blood pressure monitoring may be a prognostic marker in people with heart failure. Commentary 2. PMID- 16380031 TI - Beta-blockers may reduce mortality for elderly people with heart failure. Commentary. PMID- 16380032 TI - Non-invasive positive pressure ventilation improves outcomes for people with acute pulmonary oedema and chronic heart failure. Commentary. PMID- 16380033 TI - Decision support systems can improve clinical practice. Commentary. PMID- 16380034 TI - Nurse-led community-based care may reduce the risk of heart disease in black people at high risk. Commentary. PMID- 16380035 TI - Nurse-led clinics plus a home visit may reduce rehospitalisation for people with heart failure. Commentary 1. PMID- 16380036 TI - Nurse-led clinics plus a home visit may reduce rehospitalisation for people with heart failure. Commentary 2. PMID- 16380037 TI - Adding clopidogrel to aspirin following non-ST elevation acute coronary syndromes may be cost-effective. Commentary. PMID- 16380038 TI - Eplerenone appears cost-effective for people with heart failure after acute myocardial infarction. Commentary 1. PMID- 16380039 TI - Eplerenone appears cost-effective for people with heart failure after acute myocardial infarction. Commentary 2. PMID- 16380040 TI - Nurse-led primary care clinics for the secondary prevention of heart disease appear cost-effective compared to usual care. Commentary. PMID- 16380041 TI - There is no strong evidence that low-dose aspirin should be routinely discontinued preoperatively. Commentary. PMID- 16380042 TI - Statins and n-3 fatty acids are the best lipid-lowering interventions for reducing mortality. Commentary. PMID- 16380043 TI - Intensive lipid-lowering is no more effective than diet and low-dose lovastatin for reducing ischaemia in people with heart disease. Commentary. PMID- 16380044 TI - Here's to another great year of high quality evidence! PMID- 16380045 TI - Risk management after peripheral arterial interventions. PMID- 16380046 TI - Weight loss may not be associated with such large reductions in blood pressure as previously assumed. Commentary. PMID- 16380047 TI - Topiramate may help reduce weight and blood pressure in obese people. Commentary. PMID- 16380048 TI - Soy protein may lower blood pressure. Commentary. PMID- 16380049 TI - Multidisciplinary programmes can reduce hospitalisation and mortality in people with heart failure. Commentary. PMID- 16380050 TI - Candesartan may prevent diabetes in people with heart failure. Commentary. PMID- 16380051 TI - Reduced dose tenecteplase plus tirofiban achieves similar TIMI 3 flow to full dose tenecteplase alone in acute myocardial infarction. Commentary. PMID- 16380052 TI - Adenosine may not improve clinical outcomes for people with STEMI. Commentary. PMID- 16380053 TI - The effect of clopidogrel on infarct size at one month remains unclear for people with STEMI. Commentary 1. PMID- 16380054 TI - The effect of clopidogrel on infarct size at one month remains unclear for people with STEMI. Commentary 2. PMID- 16380055 TI - Tenecteplase followed by immediate angioplasty is more effective than tenecteplase alone for people with STEMI. Commentary. PMID- 16380056 TI - Early invasive strategy has higher short term mortality but better longer term efficacy compared to selective strategy for people with NSTEMI. Commentary 1. PMID- 16380057 TI - Early invasive strategy has higher short term mortality but better longer term efficacy compared to selective strategy for people with NSTEMI. Commentary 2. PMID- 16380058 TI - Direct comparisons of sirolimus-eluting and paclitaxel-eluting stents are needed. Commentary. PMID- 16380059 TI - Paclitaxel-eluting stents with erodable polymer appear safe. Commentary. PMID- 16380060 TI - Prasugrel may be safe for further testing in people undergoing PCI. Commentary. PMID- 16380061 TI - Invasive strategy may reduce infarct size more than conservative treatment for people with STEMI presenting within 12 to 48 hours. Commentary. PMID- 16380062 TI - Manual thrombus-aspiration is clinically feasible for people with STEMI undergoing urgent PCI. Commentary. PMID- 16380063 TI - Thrombectomy prior to PCI may improve ST-segment resolution in acute myocardial infarction. Commentary. PMID- 16380064 TI - Regular nurse follow up calls may improve time to hospitalisation, but not other outcomes, for people with heart failure. Commentary. PMID- 16380065 TI - Further studies are needed to assess the benefits of telemonitoring and telephone support for people with heart failure. Commentary. PMID- 16380066 TI - Statins may be cost effective for a wide range of subgroups. Commentary. PMID- 16380067 TI - Single and dual chamber pacemakers appear equally effective in elderly people with high grade AV block. Commentary. PMID- 16380068 TI - N-3 polyunsaturated fatty acids reduce atrial fibrillation following coronary artery bypass graft surgery. Commentary. PMID- 16380069 TI - Prayer and music, imagery, and touch therapy may not improve clinical outcomes following PCI. Commentary. PMID- 16380070 TI - [Parastomal hernias--1 (technique without transposition)]. AB - While relatively common, parastomal hernias are often well tolerated. The decision to operate and the choice of surgical technique should be well discussed taking into account the controversial results of the literature. We describe in the present article the repair techniques without stoma transposition. PMID- 16380071 TI - The effects of occupational health and safety management on work environment and health: a prospective study. AB - According to Norway's Internal Control Regulation, all companies are required to have an occupational health and safety (H&S) management system. This study investigated the effects of implementing or improving occupational H&S management on the work environment, H&S-related behaviour and musculoskeletal health of workers in small and medium-sized companies. A one-year prospective cohort study, using self-administered questionnaires, was performed among the managers and blue collar workers in 226 motor vehicle repair garages. Out of 1559 workers that responded at baseline 721 workers could be identified at follow-up. These 721 workers were included in the study. The workers in companies with improved H&S management from baseline to follow-up reported increased satisfaction with the H&S activities at the garage; improved support from management and colleagues; improved health-related support and control; and increased participation in H&S activities. PMID- 16380072 TI - Evaluation of the effectiveness of portable ceiling lifts in a new long-term care facility. AB - Researchers and health and safety practitioners have advocated replacing manual patient handling techniques with ceiling lifts in long-term care. The majority of these studies have only evaluated the impact of fixed ceiling lifts on extended care residents where the ratio of ceiling lifts to resident beds is one to one. This pre-post intervention study assesses the effectiveness of portable ceiling lifts in a new multi-level care facility on risk of patient handling injuries where the ratio of ceiling lifts to resident beds is one to six. Results indicated that staff perceived they were at significantly (p<0.05) less risk of injury when using ceiling lifts compared to manual methods. Seventy-five percent of staff preferred to use the ceiling lifts over any other method for lifting and transferring residents. Compensation costs due to patient handling decreased in the intervention facility, with a 241% increase in the comparison facility. This study demonstrates that incorporating ceiling lifts into the design of a new multi-level care facility reduced patient handling injuries and decreased perceived risk of injury among care staff. PMID- 16380073 TI - Hand-arm vibration syndrome (HAVS) and musculoskeletal symptoms in the neck and the upper limbs in professional drivers of terrain vehicles--a cross sectional study. AB - This study compares the prevalence of symptoms of Hand-arm vibration syndrome (HAVS) and musculoskeletal symptoms in the neck and the upper limbs, between professional drivers of terrain vehicles and a referent group. 769 male professional drivers of forest machines, snowmobiles, snowgroomers and reindeer herders and 296 randomly selected male referents completed a questionnaire about symptoms of HAVS and musculoskeletal symptoms in the neck and the upper limbs. They also gave information about their lifetime exposure duration driving terrain vehicles and their nicotine use. Prevalence odds ratios (POR) were determined and adjusted for age and nicotine use. Results show that there is a relation between exposure to driving terrain vehicles and some of the symptoms of HAVS (POR: 1.2 6.1). Increased odds of musculoskeletal symptoms in neck, shoulders and wrists were also found (POR 1.2-6.4), and it seemed to be related to the cumulative exposure time. PMID- 16380074 TI - Developing novel hCT derived cell-penetrating peptides with improved metabolic stability. AB - Many promising therapeutics are currently awaiting their clinical application. Due to their low capability of cell membrane crossing, these compounds do not reach their site of action. One way to overcome this problem might be the fusion of these agents to cell-penetrating peptides (CPP), which are able to shuttle various cargoes across cellular membranes. One disadvantage in using CPP in drug delivery is their low metabolic stability. The aim of our work was to increase the proteolytic resistance of the CPP hCT(9-32), a truncated C-terminal fragment of human calcitonin. Thus, we synthesised six modified N-terminally carboxyfluorescein labelled hCT(9-32) derivatives by replacing positions 12 and/or 16 of hCT(9-32) with either N-methylphenylalanine or d-phenylalanine, respectively. By using confocal laser scanning microscopy we showed that the modifications did neither affect the peptide internalisation efficiency in HeLa nor HEK 293T cells. The metabolic stability of the peptides was investigated in human blood plasma and HEK 293T cell culture supernatant. To analyse the degradation patterns, we used RP-HPLC and MALDI-TOF mass spectrometry. However, we found for all of the new derivatives high metabolic stabilities. In blood plasma, the half-lives for five of the six peptides increased compared to unmodified hCT(9-32). The degradation patterns showed a distinct stabilisation in the N-terminal part of the modified peptides, in the C-terminal part, we found some cleavage to a minor extent. Furthermore, we studied the conformation of the peptides by CD spectroscopy and demonstrated that they possess no cell toxicity. Since our metabolically more stable compounds are still able to pass the cell membrane they provide powerful tools as drug delivery vectors. PMID- 16380075 TI - The human transcription factor AP-1 is a mediator of bile acid-induced liver cell apoptosis. AB - Apoptosis induced by toxic bile acids is thought to contribute to liver injury during cholestasis. The transcription factor AP-1 is involved in the induction of apoptosis depending on stimulus and cell type. It is not known whether the major human toxic bile acid, glycochenodeoxycholic acid (GCDCA), modulates AP-1 in hepatocytes. Our data show that GCDCA (75 microM, 4 h) significantly upregulates cFos and JunB as demonstrated by microarray analysis and real-time PCR in HepG2 Ntcp hepatoma cells. GCDCA (75 microM, 4 h) also induced AP-1 activation as determined by EMSA that was most distinct after 30 min. In parallel, AP-1 transcriptional activity increased by 40% after exposure to GCDCA. Curcumin, an AP-1 inhibitor, dose-dependently reduced (1-25 microM) or completely abolished (50 microM) the apoptotic effect of GCDCA. Thus, GCDCA-induced upregulation of AP 1-dependent genes appears important for the cytotoxicity of this bile acid. PMID- 16380076 TI - An N-glycan structure correlates with pulmonary metastatic ability of cancer cells. AB - N-Glycan structures on the surface of cancer cells have diverse structures and play significant roles in metastatic process. However, little is known about their roles in organ-selective metastasis. Our study revealed that an alpha1,6 fucosylated biantennary N-glycan structure designated A2G2F is characteristic of lungs, with far more abundant expression in normal human and murine lungs than in other organs. In this study, we further examined the role of A2G2F in pulmonary metastasis. We stained metastatic cancers by alpha1,6-fucose-specific Lens culinaris agglutinin lectin and revealed that pulmonary metastatic nodules more abundantly expressed alpha1,6-fucosylated N-glycans than hepatic metastatic nodules from common primary cancers. The most specific alpha1,6-fucosylated N glycan structure in pulmonary metastatic cancer was identified to be A2G2F. Using a B16 melanoma cell metastasis model, we showed that A2G2F-rich B16 cells formed more pulmonary metastatic nodules than A2G2F-poor cells. Our results suggest that A2G2F plays a critical role in pulmonary metastasis. PMID- 16380077 TI - Introduction of alpha-hydroxymethyamino acid residues in substrate specificity P1 position of trypsin inhibitor SFTI-1 from sunflower seeds retains its activity. AB - In many complexes formed by serine proteinases and their inhibitors, the hydroxyl group provided by water molecule or by the inhibitor Ser residue is located close to the inhibitor P1-P1' reactive site. In order to investigate the role of this group, we synthesized analogues of trypsin inhibitor SFTI-1 isolated from the seeds of sunflower modified in P1 by alpha-hydroxymethylserine (HmSer) and both enantiomers of alpha-hydroxymethylvaline (HmVal). All the synthesized analogues inhibited bovine beta-trypsin and human leukocyte elastase. SFTI-1 analogues with HmVal and HmSer appear to be potent inhibitors of bovine beta-trypsin, whereas [Val5]SFTI-1 is practically inactive. Also trypsin inhibitory activity of [Ser5]SFTI-1 is significantly lower. Since the electrostatic interaction between protonated epsilon-NH2 group of the inhibitor P1 position and beta-carboxylate of trypsin Asp189 is the main driving force for interaction of both molecules, the results obtained are very interesting. We believe that these SFTI-1 analogues belong to a novel class of serine proteinase inhibitors. PMID- 16380078 TI - Low density lipoproteins inhibit the Na+/H+ antiport in human platelets via activation of p38MAP kinase. AB - Low density lipoproteins (LDL) inhibit the Na+/H+ antiport and thereby sensitize platelet towards agonist. However, mechanisms underlying the suppressing effect of LDL on Na+/H+ exchange are unclear. We here show that the lowering of intracellular pH and the suppression of the sodium propionate-induced Na+/H+ exchange in the presence of LDL are abolished by SKF86002, a selective inhibitor of p38MAP kinase (p38MAPK). The inhibitory effect of LDL on Na+/H+ exchange was mimicked by H2O2, which directly activates p38MAPK. Exposure of platelets to LDL or H2O2 led to phosphorylation of p38MAPK, its upstream regulator MAP kinase kinase 3/6 (MKK 3/6), and its downstream target heat shock protein 27 (HSP27), and this effect was abrogated in SKF86002-pretreated platelets. In addition, both LDL and H2O2 produced the SKF86002-sensitive phosphorylation of an oligopeptide encompassing p38MAPK phosphorylation sequence derived from NHE-1, a major Na+/H+ exchanger in platelets. We further show that the sensitizing effects of LDL on the thrombin-induced platelet activation, as reflected by aggregation and granule secretion, are abolished in cells pretreated with SKF86002. We conclude that activation of p38MAPK is required for the inhibitory effect of LDL on Na+/H+ antiport and thereby for LDL-dependent sensitization in human platelets. PMID- 16380079 TI - Visualization of the endocytic pathway in the filamentous fungus Aspergillus oryzae using an EGFP-fused plasma membrane protein. AB - Endocytosis is an important process for cellular activities. However, in filamentous fungi, the existence of endocytosis has been so far elusive. In this study, we used AoUapC-EGFP, the fusion protein of a putative uric acid-xanthine permease with enhanced green fluorescent protein (EGFP) in Aspergillus oryzae, to examine whether the endocytic process occurs or not. Upon the addition of ammonium into the medium the fusion protein was internalized from the plasma membrane. The internalization of AoUapC-EGFP was completely blocked by sodium azide, cold, and cytochalasin A treatments, suggesting that the internalization possesses the general features of endocytosis. These results demonstrate the occurrence of endocytosis in filamentous fungi. Moreover, we discovered that the endosomal compartments appeared upon the induction of endocytosis and moved in a microtubule-dependent manner. PMID- 16380080 TI - Elevated expression and potential roles of human Sp5, a member of Sp transcription factor family, in human cancers. AB - In this report, we describe the expression and function of human Sp5, a member of the Sp family of zinc finger transcription factors. Like other family members, the Sp5 protein contains a Cys2His2 zinc finger DNA binding domain at the C terminus. Our experiments employing Gal4-Sp5 fusion proteins reveal multiple transcriptional domains, including a N-terminal activity domain, an intrinsic repressive element, and a C-terminal synergistic domain. Elevated expression of Sp5 was noted in several human tumors including hepatocellular carcinoma, gastric cancer, and colon cancer. To study the effects of the Sp5 protein on growth properties of human cancer cells and facilitate the identification of its downstream genes, we combined an inducible gene expression system with microarray analysis to screen for its transcriptional targets. Transfer of Sp5 into MCF-7 cells that expressed no detectable endogenous Sp5 protein elicited significant growth promotion activity. Several of the constitutively deregulated genes have been associated with tumorigenesis (CDC25C, CEACAM6, TMPRSS2, XBP1, MYBL1, ABHD2, and CXCL12) and Wnt/beta-Catenin signaling pathways (BAMBI, SIX1, IGFBP5, AES, and p21WAF1). This information could be utilized for further mechanistic research and for devising optimized therapeutic strategies against human cancers. PMID- 16380081 TI - Angiotensin II promotes the proliferation of activated pancreatic stellate cells by Smad7 induction through a protein kinase C pathway. AB - Activated pancreatic stellate cells (PSCs) play major roles in promoting pancreatic fibrosis. We previously reported that angiotensin II (Ang II) enhances activated PSC proliferation through EGF receptor transactivation. In the present study, we elucidated a novel intracellular mechanism by which Ang II stimulates cellular proliferation. TGF-beta1 inhibits activated PSC proliferation via a Smad3 and Smad4-dependent pathway in an autocrine manner. We demonstrated that Ang II inhibited TGF-beta1-induced nuclear accumulation of Smad3 and Smad4. Furthermore, Ang II rapidly induced inhibitory Smad7 mRNA expression. Adenovirus mediated Smad7 overexpression inhibited TGF-beta1-induced nuclear accumulation of Smad3 and Smad4, and potentiated activated PSC proliferation. PKC inhibitor Go6983 blocked the induction of Smad7 mRNA expression by Ang II. In addition, 12 O-tetradecanoyl-phorbol 13-acetate, a PKC activator, increased Smad7 mRNA expression. These results suggest that Ang II enhances activated PSC proliferation by blocking autocrine TGF-beta1-mediated growth inhibition by inducing Smad7 expression via a PKC-dependent pathway. PMID- 16380082 TI - Single-point mutations of hepatitis C virus NS3 that impair p53 interaction and anti-apoptotic activity of NS3. AB - The N-terminal domain of NS3 of hepatitis C virus (HCV) possesses serine protease activity, which is essential for virus replication. This portion is also implicated in malignant transformation of hepatocytes. We previously demonstrated that an N-terminal portion of NS3 formed a complex with the tumor suppressor p53 and suppressed actinomycin D-induced apoptosis. We report here that single-point mutations of NS3 at position 106 from Leu to Ala (L106A), and position 43 from Phe to Ala (F43A) to a lesser extent, significantly impaired complex formation with p53. Moreover, the L106A mutation impaired an otherwise more distinct anti apoptotic activity of NS3. F43A and L106A mutations also inhibited serine protease activity of NS3. These results collectively suggest the possibility that Leu106 and Phe43 are involved in p53 interaction and serine protease activity, and therefore, can be a good target for certain low-molecular-weight compound(s) to inhibit both oncogenic and replicative abilities of HCV. PMID- 16380083 TI - Dicer and positive charge of proteins decrease the stability of RNA containing the AU-rich element of GM-CSF. AB - AU-rich elements (AREs) in the 3'-untranslated region of mRNAs promote rapid decay of the mRNAs for certain cytokines, including that encoding granulocyte macrophage colony-stimulating factor (GM-CSF). We show that an RNA molecule based on the ARE of GM-CSF mRNA is cleaved between U and A residues in the presence of bovine serum albumin of which cleavage effect is attenuated by acetylation. Furthermore, the expression of RNA molecule containing the ARE of GM-CSF mRNA in human cell lines was increased by inhibition of histone deacetylase activity and attenuation of Dicer expression. These findings suggest that degradation of mRNAs containing an ARE might be regulated by positive charge of polypeptides and Dicer. PMID- 16380084 TI - Interaction of bovine serum amine oxidase with the polyamine oxidase inactivator MDL 72527. AB - MDL 72527 was considered a selective inhibitor of FAD-dependent polyamine oxidases. In the present communication, we demonstrate that MDL 72527 inactivates bovine serum amine oxidase, a copper-containing, TPQ-enzyme, time-dependently at 25 degrees C. In striking contrast, the enzyme remained active after incubation with excessive MDL 72527 at 37 degrees C, even after 70 h of incubation. Inactivation of BSAO with MDL 72527 at 25 degrees C did not involve the cofactor, as was shown by spectroscopy and by reaction with phenylhydrazine. Docking of MDL 72527 is difficult, owing to its size and two lipophilic moieties, and it has been shown that minor changes in reaction rate of substrates cause major changes in K(m) and k(cat)/K(m). We hypothesise that subtle conformational changes between 25 and 37 degrees C impair MDL 72527 from productive binding and prevent the nucleophilic group from reacting with the double bond system. PMID- 16380085 TI - Brain magnetic resonance study of Mecp2 deletion effects on anatomy and metabolism. AB - Rett syndrome, a neurodevelopmental X-linked disorder, represents the most important genetic cause of severe mental retardation in the female population and results from a mutation in the gene encoding methyl-CpG-binding protein 2 (MECP2). We report here the first characterization of Mecp2-null mice, by in vivo magnetic resonance imaging and spectroscopy, delineating the cerebral phenotype associated with the lack of Mecp2. We performed a morphometric study that revealed a size reduction of the whole brain and of structures involved in cognitive and motor functions (cerebellum and motor cortex). Significant metabolic anomalies, including reduced N-acetylaspartate, myo-inositol, and glutamine plus glutamate, and increased choline levels were evidenced. These findings indicate that not only neuronal but also glial metabolism is affected in Mecp2-null mice. Furthermore, we uncovered an important reduction of brain ATP level, a hitherto undetected anomaly of energy metabolism that may reflect and contribute to cerebral injury and dysfunction. PMID- 16380086 TI - G protein activation is prerequisite for functional coupling between Galpha/Gbetagamma and tubulin/microtubules. AB - Heterotrimeric G proteins participate in signal transduction by transferring signals from cell surface receptors to intracellular effector molecules. Interestingly, recent results suggest that G proteins also interact with microtubules and participate in cell division and differentiation. It has been shown earlier that both alpha and betagamma subunits of G proteins modulate microtubule assembly in vitro. Since G protein activation and subsequent dissociation of alpha and betagamma subunits are necessary for G proteins to participate in signaling processes, here we asked if similar activation is required for modulation of microtubule assembly by G proteins. We reconstituted Galphabetagamma heterotrimer from myristoylated-Galpha and prenylated-Gbetagamma, and found that the heterotrimer blocks Gi1alpha activation of tubulin GTPase and inhibits the ability of Gbeta1gamma2 to promote in vitro microtubule assembly. Results suggest that G protein activation is required for functional coupling between Galpha/Gbetagamma and tubulin/microtubules, and supports the notion that regulation of microtubules is an integral component of G protein mediated signaling. PMID- 16380087 TI - Effect of the in situ electrochemical oxidation on the pigment-protein arrangement and energy transfer in light-harvesting complex from Rhodobacter sphaeroides 601. AB - The oxidation of bacteriochlorophylls (BChls) in peripheral light-harvesting complexes (LH2) from Rhodobacter sphaeroides was investigated by spectroelectrochemistry of absorption, fluorescence emission, and femtosecond (fs) pump-probe, with the aim obtaining information about the effect of in situ electrochemical oxidation on the pigment-protein arrangement and energy transfer within LH2. The experimental results revealed that: (a) the generation of the BChl radical cation in both B800 and B850 rings dramatically induced bleaching of the characteristic absorption in the NIR region and quenching of the fluorescence emission from the B850 ring for the electrochemical oxidized LH2; (b) the BChl B850 radical cation might act as an additional channel to compete with the unoxidized BChl-B850 molecules for rapidly releasing the excitation energy, however the B800-B850 energy transfer rate remained almost unchanged during the oxidation process. PMID- 16380088 TI - Mutations in p53 cDNA sequence introduced by retroviral vector. AB - The high mutation rates of retroviruses are a potential problem with retroviral vectors. We studied the mutation rates and spectra of p53 sequences transduced with a retroviral vector in a cancer gene therapy model. When p53-deficient H358 non-small cell lung cancer cells were treated with a retroviral vector carrying normal p53 cDNA, most of transduced cells were killed by apoptosis. However, a small number of clones escaped p53-mediated apoptosis. We examined the p53 cDNA structure in these resistant clones. PCR-based analysis showed that 88/102 clones had detectable mutations in p53, including gross rearrangements, deletions/insertions, and base substitutions. To study the mutation rate of the p53 sequence in all transduced clones, the retroviral vector containing the non functional p53 gene and the Neo-resistant marker gene was introduced into H358 cells. Only one of 95 isolated clones showed a base substitution. These results indicate that the mutation rate of p53 is not particularly high, but there is a significant risk that cancer cells will resist p53 gene therapy as a result of retroviral replication errors. PMID- 16380089 TI - Clinical and molecular analysis of a four-generation Chinese family with aminoglycoside-induced and nonsyndromic hearing loss associated with the mitochondrial 12S rRNA C1494T mutation. AB - We report here the clinical, genetic, and molecular characterization of a four generation Chinese family with aminoglycoside-induced and nonsyndromic hearing loss. Five of nine matrilineal relatives had aminoglycoside-induced hearing loss. These matrilineal relatives exhibited variable severity and audiometric configuration of hearing impairment, despite sharing some common features: being bilateral and having sensorineural hearing impairment. Sequence analysis of mitochondrial DNA (mtDNA) in the pedigree identified 16 variants and the homoplasmic 12S rRNA C1494T mutation, which was associated with hearing loss in the other large Chinese family. In fact, the occurrence of the C1494T mutation in these genetically unrelated pedigrees affected by hearing impairment strongly indicated that this mutation is involved in the pathogenesis of aminoglycoside induced and nonsyndromic hearing loss. However, incomplete penetrance of hearing loss indicated that the C1494T mutation itself is not sufficient to produce a clinical phenotype but requires the involvement of modifier factors for the phenotypic expression. Those mtDNA variants, showing no evolutional conservation, may not have a potential modifying role in the pathogenesis of the C1494T mutation. However, nuclear background seems to contribute to the phenotypic variability of matrilineal relatives in this family. Furthermore, aminoglycosides modulate the expressivity and penetrance of deafness associated with the C1494T mutation in this family. PMID- 16380090 TI - Lipopolysaccharide prevents apoptosis induced by brefeldin A, an endoplasmic reticulum stress agent, in RAW 264.7 cells. AB - The effect of lipopolysaccharide (LPS) on the cell death induced by endoplasmic reticulum (ER) stress agents in RAW 264.7 cells was studied. LPS prevented the cell death by brefeldin A, but not thapsigargin and tunicamycin. CpG DNA as well as LPS prevented brefeldin A-induced cell death whereas tumor necrosis factor alpha or interferon-gamma did not. Brefeldin A-induced cell death was mediated with apoptotic cell death and it was significantly inhibited by LPS. LPS abolished the activation of ER stress-related caspases, such as caspases 1, 3, and 4. LPS prevented brefeldin A-induced morphological changes in RAW 264.7 cells. Further, LPS prevented brefeldin A-induced Golgi dispersion. Therefore, LPS was suggested to diminish the stress of ER/Golgi complexes induced by brefeldin A and inhibit apoptosis. The preventive action of LPS on brefeldin A induced apoptosis is discussed. PMID- 16380091 TI - ERp29 is an essential endoplasmic reticulum factor regulating secretion of thyroglobulin. AB - ERp29 is a ubiquitously expressed endoplasmic reticulum (ER) protein, which is found in the folding complexes of several secretory proteins in the ER. In our previous work, it was suggested that ERp29 function is critical for the folding/secretion of thyroglobulin (Tg), a major secretory product of thyroid cells. Current work is an attempt to substantiate this assumption by answering the question whether the secretion of Tg can be regulated through the manipulation of ERp29 expression in the FRTL-5 rat thyroid cells. Indeed, transient overexpression of ERp29 resulted in twofold enhancement of the Tg secretion whereas the RNAi-mediated ERp29 silencing led to the attenuation of the Tg export. Mutational analysis has suggested two loci that might be involved in the ERp29-Tg interactions: the interdomain linker including Cys157, an amino acid, which is important for the structural integrity of the C-terminal domain and an uncharged surface on the N-terminal domain flanked by Tyr64 and Gln70. PMID- 16380092 TI - Multiple unfolding states of glutathione transferase from Physa acuta (Gastropoda [correction of Gastropada]: Physidae). AB - The equilibrium unfolding of the major Physa acuta glutathione transferase isoenzyme (P. acuta GST(3)) has been performed using guanidinium chloride (GdmCl), urea, and acid denaturation to investigate the unfolding intermediates. Protein transitions were monitored by intrinsic fluorescence. The results indicate that unfolding of P. acuta GST(3) using GdmCl (0-3.0M) is a multistep process, i.e., three intermediates coexist in equilibrium. The first intermediate, a partially dissociated dimer, exists at low GdmCl concentration (approximately at 0.7M). At 1.2M GdmCl, a dimeric intermediate with a compact structure was observed. This intermediate undergoes dissociation into structural monomers at 1.75M of GdmCl. The monomeric intermediate started to be completely unfolding at higher GdmCl concentrations (>1.8M). Unfolding using urea (0-7.0M) and acid-induced structures as well as the fluorescence of 8-anilino-1 naphthalenesulfonate in the presence of different GdmCl concentrations confirmed that the unfolding is a multistep process. At concentrations of GdmCl or urea less than the midpoints or at the midpoint pH (pH 4.2-4.6), the unfolding transition is protein concentration independent and involved a change in the subunit tertiary structure yielding a partially active dimeric intermediate. The binding of glutathione to the enzyme active site stabilizes the native dimeric state. PMID- 16380093 TI - Decorin binds myostatin and modulates its activity to muscle cells. AB - Myostatin, a member of TGF-beta superfamily of growth factors, acts as a negative regulator of skeletal muscle mass. The mechanism whereby myostatin controls the proliferation and differentiation of myogenic cells is mostly clarified. However, the regulation of myostatin activity to myogenic cells after its secretion in the extracellular matrix (ECM) is still unknown. Decorin, a small leucine-rich proteoglycan, binds TGF-beta and regulates its activity in the ECM. Thus, we hypothesized that decorin could also bind to myostatin and participate in modulation of its activity to myogenic cells. In order to test the hypothesis, we investigated the interaction between myostatin and decorin by surface plasmon assay. Decorin interacted with mature myostatin in the presence of concentrations of Zn(2+) greater than 10microM, but not in the absence of Zn(2+). Kinetic analysis with a 1:1 binding model resulted in dissociation constants (K(D)) of 2.02x10(-8)M and 9.36x10(-9)M for decorin and the core protein of decorin, respectively. Removal of the glycosaminoglycan chain by chondroitinase ABC digestion did not affect binding, suggesting that decorin could bind to myostatin with its core protein. Furthermore, we demonstrated that immobilized decorin could rescue the inhibitory effect of myostatin on myoblast proliferation in vitro. These results suggest that decorin could trap myostatin and modulate its activity to myogenic cells in the ECM. PMID- 16380094 TI - Disruption of caveolin-1 leads to enhanced nitrosative stress and severe systolic and diastolic heart failure. AB - Although caveolin-1 is not expressed in cardiomyocytes, this protein is assumed to act as a key regulator in the development of cardiomyopathy. In view of recent discordant findings we aimed to elucidate the cardiac phenotype of independently generated caveolin-1 knockout mice (cav-1(-/-)) and to unveil causative mechanisms. Invasive hemodynamic measurements of cav-1(-/-) show a severely reduced systolic and diastolic heart function. Additionally, genetic ablation of caveolin-1 leads to a striking biventricular hypertrophy and to a sustained eNOS hyperactivation yielding increased systemic NO levels. Furthermore, a diminished ATP content and reduced levels of cyclic AMP in hearts of knockout animals were measured. Taken together, these results indicate that genetic disruption of caveolin-1 is sufficient to induce a severe biventricular hypertrophy with signs of systolic and diastolic heart failure. Collectively, our findings suggest a causative role of a sustained nitrosative stress in the development of the pronounced cardiac impairment. PMID- 16380095 TI - 3,3'-Diindolylmethane downregulates pro-survival pathway in hormone independent prostate cancer. AB - Epidemiological evidences suggest that the progression and promotion of prostate cancer (CaP) can be modulated by diet. Since all men die with prostate cancer rather than of the disease, it is of particular interest to prevent or delay the progression of the disease by chemopreventive strategies. We have been studying the anticancer properties of compounds present in cruciferous vegetables such as indole-3-carbinol (I3C). Diindolylmethane (DIM) is a dimer of I3C that is formed under acidic conditions and unlike I3C is more stable with higher anti-cancer effects. In the present report, we demonstrate that DIM is a potent anti proliferative agent compared to I3C in the hormone independent DU 145 CaP cells. The anti-prostate cancer effect is mediated by the inhibition of the Akt signal transduction pathway as DIM, in sharp contrast to I3C, induces the downregulation of Akt, p-Akt, and PI3 kinase. DIM also induced a G1 arrest in DU 145 cells by flow cytometry and downstream concurrent inhibition of cell cycle parameters such as cyclin D1, cdk4, and cdk6. Our data suggest a need for further development of DIM, as a chemopreventive agent for CaP, which justifies epidemiological evidences and molecular targets that are determinants for CaP dissemination/progression. The ingestion of DIM may benefit CaP patients and reduce disease recurrence by eliminating micro-metastases that may be present in patients who undergo radical prostatectomy. PMID- 16380096 TI - Prenatal exposure to bisphenol A impairs sexual differentiation of exploratory behavior and increases depression-like behavior in rats. AB - Perinatal exposure to bisphenol A (BPA, 0.1 and 1 ppm in drinking water applied to mother rats for 6 weeks) has been shown to impair the sexual differentiation in exploratory behavior, but the exact critical period of this disrupting effect is still unknown. In this study, we examined the effects of prenatal exposure to BPA (0.1 ppm in drinking water applied to dams during the final week of pregnant) on emotional and learning behaviors in addition to exploratory behavior. Estimated daily intake was 15 microg/kg/day, below the reference dose (RfD) in the United States and the daily tolerable intake (TDI) in Japan (50 microg/kg/day). The rats were successively tested in open-field test, elevated plus maze test, passive avoidance test and forced swimming test during development from 6 to 9 weeks of juvenile period. Prenatal exposure to BPA mainly affected male rats and abolished sex differences in rearing behavior in the open field test and struggling behavior in the forced swimming test. BPA increased the immobility of male rats in the forced swimming test. The avoidance learning and behaviors in the elevated plus maze were not affected. The present study demonstrates that male rats at the final week of prenatal period are sensitive to BPA, which impairs sexual differentiation in rearing and struggling behavior and facilitate depression-like behavior. PMID- 16380097 TI - Changes of neuronal activity in areas CA1 and CA3 during anoxia and normoxic or hyperoxic reoxygenation in juvenile rat organotypic hippocampal slice cultures. AB - In neonates, asphyxia is usually followed by hyperoxic treatment. In order to study whether hyperoxic reoxygenation might cause additional impairment of neuronal function, we subjected organotypic hippocampal slice cultures of juvenile rats (7 DIV, P6-8) to 30 min anoxia followed by 60 min hyperoxic or normoxic reoxygenation (95% or 19% O2, respectively). Spontaneous and evoked field potentials as well as [Ca2+]o were recorded in the pyramidal layer of area CA1 or area CA3. In area CA1, 30 min of anoxia led to decline of evoked field potential amplitudes by on average 67% and to profound changes in field potential characteristics and Ca2+ homeostasis which were not related to outcome after reoxygenation. Hyperoxic reoxygenation resulted first in a fast recovery of the field potential amplitude to 82% of the control value and then, in 75% of slice cultures, in a large negative field potential shift accompanied by a prolonged decrease of [Ca2+]o and loss of excitability outlasting the experiment. Recovery of field potential amplitude under normoxic conditions stayed poor, with a first increase to 51% and a second decrease to 22%. In contrast, field potential amplitude in area CA3 recovered to 80% of the initial amplitude, irrespective of the reoxygenation mode. The selective loss of function during hyperoxic reoxygenation in area CA1 might be a first sign of neuronal injury that we observed 1 h after end of hyperoxic reoxygenation in a previous study. Whether the poor outcome after normoxic reoxygenation would favour long-term recovery remains to be determined. PMID- 16380098 TI - Prenatal treatment with picrotoxin promotes heterotypical sexual behavioral and neurochemical changes in male rat offspring. AB - The effects of maternal prenatal exposure to picrotoxin (0.75 mg/kg S.C. days 16 19 of pregnancy) in male rat offspring were observed. Adult sexually experienced and inexperienced animals were evaluated for heterotypical sexual behavior, as well as the testosterone plasma levels and striatal neurotransmitters. In relation to sexual behavior and analysis of sexual organs, the results showed that animals treated with picrotoxin exhibited a more intense reproductive behavior, and this could be expressed by a significant decrease in the number of mounts and intromissions and increase in the numbers of ejaculation, showed that these males are most motivate for sexual behavior. Testosterone levels as well as weight for sexual organs did not differ from control group. The neurochemical analysis showed that picrotoxin did not alter DA, 5-HT, 5-HIAA and GABA in animals. The DOPAC/DA and HVA/DA relation showed that the treatment increased the DA system activity in animals sexually experienced, as well as promote a decrease in 5-HT/5-HIAA relation, that is known was an inhibitory neurotransmitter system, blockade a male sexual behavior. There are no alterations observed in GABA levels. It's could be explained by suggests that picrotoxin modification DA system activity through GABAergic system, permitting that DA system to be freely active and facilitate the heterotypical behavior of male rats. These results show that the maternal prenatal exposure to picrotoxin produced changes in the neurochemical and sexual behavior of the adult male rats. Also previous heterotypical experience leads to changes in biogenic amine concentrations in these animals. PMID- 16380099 TI - Milk-derived lactoferrin may block tolerance to morphine analgesia. AB - Lactoferrin (LF) is a multifunctional protein that is widely found in milk, blood, and other biological fluids. In the present study, we investigated the possibility that LF may block a tolerance to morphine-induced analgesia in the mouse. The nociceptive effect of bovine milk-derived LF (bLF) was estimated in the mouse tail-flick test. Although an intraperitoneal (100 mg/kg) or an oral (300 mg/kg) administration of bLF did not show remarkable analgesia, a combination with intraperitoneal administration of morphine (3 mg/kg) strikingly enhanced morphine-induced analgesia. Moreover, repeated administration of morphine at doses of 3 mg/kg (ip) or 5 mg/kg (ip) caused a tolerance to the morphine on the 5th or 7th day, respectively. In contrast, the combination of bLF (100 mg/kg, ip) with morphine (3 mg/kg, ip) retarded the development of tolerance to the 9th day, although bLF did not show any effect on the mice that had obtained tolerance to morphine. Furthermore, the potentiative effect of bLF was partially blocked by pre-treatment with N(G)-nitro-L-arginine methyl ester (L NAME), a nonselective nitric oxide synthase (NOS) inhibitor, and completely blocked by 7-nitroindazole (7-NI), a selective neuronal NOS (nNOS) inhibitor. Methylene blue (MB), a guanylate cyclase (GC) inhibitor, also dose-dependently prevented the potentiative effect of bLF. These results suggest that bLF selectively activates nNOS and then accelerates NO production. The increased NO in turn modulates the GC activity and finally enhances the endogenous opioid system via cyclic guanosine monophosphate production. We conclude that bLF may block the development of tolerance to morphine in mice, possibly via the selective activation of nNOS. PMID- 16380100 TI - GDNF family ligands display distinct action profiles on cultured GABAergic and serotonergic neurons of rat ventral mesencephalon. AB - Glial-cell-line-derived neurotrophic factor (GDNF), neurturin (NRTN), artemin (ARTN) and persephin (PSPN), known as the GDNF family ligands (GFLs), influence the development, survival and differentiation of cultured dopaminergic neurons from ventral mesencephalon (VM). Detailed knowledge about the effects of GFLs on other neuronal populations in the VM is essential for their potential application as therapeutic molecules for Parkinson's disease. Hence, in a comparative study, we investigated the effects of GFLs on cell densities and morphological differentiation of gamma-aminobutyric acid-immunoreactive (GABA-ir) and serotonin ir (5-HT-ir) neurons in primary cultures of E14 rat VM. We observed that all GFLs [10 ng/ml] significantly increased GABA-ir cell densities (1.6-fold) as well as neurite length/neuron. However, only GDNF significantly increased the number of primary neurites/neuron, and none of the GFLs affected soma size of GABA-ir neurons. In contrast, only NRTN treatment significantly increased 5-HT-ir cells densities at 10 ng/ml (1.3-fold), while an augmentation was seen for GDNF and PSPN at 100 ng/ml (2.4-fold and 1.7-fold, respectively). ARTN had no effect on 5 HT-ir cell densities. Morphological analysis of 5-HT-ir neurons revealed a significant increase of soma size, number of primary neurites/neuron and neurite length/neuron after GDNF exposure, while PSPN only affected soma size, and NRTN and ARTN failed to exert any effect. In conclusion, we identified GFLs as effective neurotrophic factors for VM GABAergic and serotonergic neurons, demonstrating characteristic individual action profiles emphasizing their important and distinct roles during brain development. PMID- 16380101 TI - Hemispheric specialization for language: Brain volume matters. AB - Increasing brain volume may impose constraints, through longer information transfer delays, on the distributed networks supporting language. Here, we assessed the relative effects of brain volume and other putative predictors of the functional variability of perisylvian language areas, as probed with PET, during both a language comprehension and a language production task. In the case of language comprehension (story listening), a linear combination of planum temporale surface, brain volume and handedness could explain almost 60% of the functional asymmetry observed in the perisylvian area. Without brain volume, the goodness of fit was significantly decreased (39%, P < 0.05), and furthermore, the effect of handedness was not detected anymore. This was due to the fact that in our sample, left-handers (n = 12) had a significantly larger brain volume as compared to right-handers (n = 8, P = 0.03). As for language production (verb generation), brain volume and the planum temporale also played a role. However, in this case, the main predictor of functional variability was handedness, where a greater degree of right-handedness was associated with larger activation of left inferior frontal regions. Depending on the language component of interest, these results support different (yet compatible) theories on hemispheric specialization. Left specialization for comprehension could be attributed to the constraints of processing speech stimuli, while a gestural origin of language is mostly supported by the relation we observed between left specialization for production and right-handedness. PMID- 16380102 TI - Analysis of mouse brain using a clinical 1.5 T scanner and a standard small loop surface coil. AB - With increasing numbers of in vivo experiments in the field of neuroscience, the interest in methods for in vivo imaging of animal brains as small as those of mice has increased. Because highly specialized small bore scanners with high field strengths are not commonly available, clinical magnetic resonance imaging (cMRI) scanners have been used in the past to image rat and more recently also mouse brains in combination with specifically developed RF coils. These studies have demonstrated that imaging of small animal brains is feasible, and that tumor volumes measured by cMRI correlate well with histological tumor volume analysis. This protocol describes the cMRI settings at 1.5 T for imaging of mouse brain with resolutions up to 120 x 120 microm using an inexpensive, commercially available small loop surface coil. This allows easy establishment of a small animal MRI facility without the need for cost intensive dedicated small animal scanners or special custom made coils. In this study, we demonstrate high resolution imaging of intracranial xenografts in a mouse glioma model and monitor the treatment effect of external field irradiation by cMRI. PMID- 16380103 TI - Length and protein kinase A modulations of myocytes in cardiac myosin binding protein C-deficient mice. AB - OBJECTIVE: Beta-adrenergic stimulation modulates cardiac contractility through protein kinase A (PKA), which phosphorylates proteins such as troponin I (cTnI) and C-protein (cMyBP-C). The relative contributions of cTnI and cMyBP-C to the regulation of myofilament Ca(2+) sensitivity are still controversial because of difficulty in targeting specific protein phosphorylation. Recently, impaired relaxation was found in cMyBP-C-deficient mice (KO) in vivo under basal conditions and after beta-adrenergic stimulation. The goal of this study was to analyse the length-dependent and PKA-dependent modulations of the cardiac contractile machinery in a mouse model lacking cMyBP-C. METHODS: In the present work, we studied the PKA effect on myofilament Ca(2+) sensitivity of left ventricular skinned myocytes isolated from 5-week- and 55-week-old wild-type (WT) and cMyBP-C knockout (KO) mice at 1.9 and 2.3 mum sarcomere lengths (SL). The cTnI content and phosphorylation status were examined by Western blot analysis. RESULTS: Without PKA stimulation and at the shorter SL, Ca(2+) sensitivity was higher in KO compared to WT. The difference disappeared at the longer SL. No difference in passive tension or maximal active tension was observed. PKA stimulation induced a desensitization of WT myofilaments at both SL but had almost no effect in KO myofilaments despite similar levels of cTnI phosphorylation. We also observed expression of slow skeletal TnI in KO animals that was not correlated with the PKA effects. CONCLUSION: The results suggest that cMyBP-C contributes to the regulation of cardiac contraction at short sarcomere length and that myofilament desensitization induced by PKA requires the presence of cMyBP-C and does not depend only upon TnI phosphorylation. PMID- 16380104 TI - Evidence for the role of lipid peroxides on glycation of hemoglobin and plasma proteins in non-diabetic asthma patients. AB - BACKGROUND: Collective evidences reveal that malondialdehyde (MDA), reduced glutathione (GSH) and ascorbic acid can modulate protein glycation. We investigated the concentrations of MDA, GSH, ascorbic acid and protein glycation in asthma patients to delineate the possible association among these parameters. METHODS: Blood was collected from 18 asthma patients and 16 age and sex matched control subjects. Glycated hemoglobin (HbA1C), GSH, MDA, vitamin C, fructosamine and glucose were assessed in both groups. The effect of H2O2 on glycation of hemoglobin was studied by incubating normal healthy erythrocytes with either 5 or 50 mmol/l glucose concentration. RESULTS: Plasma of asthma patients revealed significantly higher concentrations of lipid peroxides and fructosamine concentrations than the matched controls. Glycated hemoglobin concentrations were also found to be significantly increased. Ascorbic acid and GSH concentrations were decreased significantly in the test group when compared with the healthy control group. When the effects of fasting glucose, GSH and ascorbic acid on the concentrations of HbA1C and fructosamine were refuted by partial correlation analysis, MDA was found to be a significant determinant of HbA1c and fructosamine in patients with asthma. The in vitro model with human erythrocytes showed an enhancement of protein glycation by H2O2. CONCLUSION: An increased glycation of proteins was found in asthma patients. These data also support the premise that lipid peroxides per se do have a role to play in glycation of hemoglobin and plasma proteins. PMID- 16380105 TI - Influence of T-786C polymorphism on the promoter activity of eNOS. PMID- 16380107 TI - Infant music perception: domain-general or domain-specific mechanisms? AB - We review the literature on infants' perception of pitch and temporal patterns, relating it to comparable research with human adult and non-human listeners. Although there are parallels in relative pitch processing across age and species, there are notable differences. Infants accomplish such tasks with ease, but non human listeners require extensive training to achieve very modest levels of performance. In general, human listeners process auditory sequences in a holistic manner, and non-human listeners focus on absolute aspects of individual tones. Temporal grouping processes and categorization on the basis of rhythm are evident in non-human listeners and in human infants and adults. Although synchronization to sound patterns is thought to be uniquely human, tapping to music, synchronous firefly flashing, and other cyclic behaviors can be described by similar mathematical principles. We conclude that infants' music perception skills are a product of general perceptual mechanisms that are neither music- nor species specific. Along with general-purpose mechanisms for the perceptual foundations of music, we suggest unique motivational mechanisms that can account for the perpetuation of musical behavior in all human societies. PMID- 16380106 TI - Proapoptotic and antitumor activities of the HMG-CoA reductase inhibitor, lovastatin, against Dalton's lymphoma ascites tumor in mice. AB - BACKGROUND: Diet rich in fat have a clear effect on the tumor incidence in humans. Increased level of lipid peroxidation were found in colon, liver, breast and kidney carcinogenesis. Although the beneficial effects of statins for cardiovascular diseases are well established, their importance in the area of cancer therapeutics has recently gained recognition. Many studies of lovastatin in in vitro systems and experimental animals have been reported as an effective antitumor agent. However, phase I/II clinical trials in cancer patients demonstrated a minor to non-significant responses. Hence more studies in different tumor models using doses corresponding to that used to reduce lipid in human are required to support the antitumor activity. METHODS: The antitumor activity was evaluated using Daltons' Lymphoma Ascites (DLA) cell line-induced ascites tumor model in mice. Proapoptotic activity was evaluated in DLA cell line induced ascites animals after the treatment of lovastatin. Apoptosis was analyzed morphologically by staining with Giemsa and biochemically by observing the laddering of DNA in agarose gel electrophoresis. In vitro cytotoxic activity of lovastatin was studied by trypan blue dye exclusion method. Lipid peroxidation inhibiting activity was demonstrated in Fe2+-ascorbate induced rat whole liver homogenate. RESULTS: Lovastatin dose dependently inhibited the ascites tumor growth at 4 and 16 mg/kg body wt (p.o). The percentage increase in life span (%ILS) in the 16 mg/kg treated group was 61.8% (P<0.01). Single dose of lovastatin (16 mg/kg body wt, p.o) was also effective to accelerate the apoptosis in the ascites tumor bearing mice that was evident from the multiple fragmentation of DNA in gel electrophoresis. Further the morphological analysis of DLA cells aspirated from the lovastatin treated animals showed a significant (P<0.01) increase of apoptotic cells (15.5+/-3%) than the control animals (6.5+/ 1%). Concentration of lovastatin required for the 50% of the cytotoxicity was 37+/-5 microg/ml. Lovastatin at its low concentrations were effective to inhibit lipid peroxidation. CONCLUSIONS: The antitumor activity of lovastatin against the ascites tumor is due to its proapoptotic and cytotoxic activities. Lovastatin at low concentrations inhibited Fe2+ induced lipid peroxidation in in vitro system. The proapoptotic and lipid peroxidation inhibiting activities of the lipid lowering drug lovastatin may further suggest its possible therapeutic use as a cancer chemopreventive agent. PMID- 16380108 TI - The role of attention in the facilitation effect and another "inhibition of return". AB - The present study investigated how anticipation of a target's appearance affects human attention to gaze cues provided by a schematic face. Subjects in a 'catch' group received a high number of 'catch' trials, in which no target stimulus appeared. Subjects in the control group did not receive any catch trials. As in previous studies, both groups showed a facilitation effect to the cued location during shorter stimulus onset asynchrony (SOA). In both groups, an analysis of eye movements confirmed that subjects' eyes remained on the fixation point, ruling out the possibility that the facilitation effect was due to shifting eye movements (saccades) as opposed to a shift in covert attention. But while the control group's response time (RT) decreased as SOA increased, the catch group's RT had a U-shaped pattern and the facilitation effect to the cued location was reversed at the longest SOA (1005 ms). These results suggest that subjects in the catch group disengaged their attention during long SOAs because they expected the trial to be a catch trial. This disengagement of attention during long SOAs results in a delay before attention could be re-focused to the previous location regardless of the cue validity ["IOR (inhibition of return)"-like-phenomenon]. Unlike the conventional IOR, we suggest that this "IOR"-like phenomenon caused by an unpredictive central gaze cue is likely to be mediated by an endogenous mechanism. PMID- 16380109 TI - Reduced fertility of female mice lacking CD81. AB - In somatic cells, the tetraspanins CD81 and CD9 associate with each other, with additional tetraspanins and with non-tetraspanin molecules to form proteolipidic complexes. Here we show that CD81 is expressed on the surface of oocytes where it associates with tetraspanin-enriched membrane structures. A major CD9 and CD81 partner, CD9P-1, is also expressed by oocytes. Deletion of CD81 gene in mice results in a 40% reduction of female fertility. In vitro insemination indicated that this infertility is due to a deficiency of oocytes to fuse with sperm. While the fertility of CD9-/- mice is severely but not completely impaired, double knock-out CD9-/- CD81-/- mice were completely infertile indicating that CD9 and CD81 play complementary roles in sperm-egg fusion. Finally, a fraction of CD9 was transferred from CD81-/- oocytes to sperm present in the perivitelline space indicating that the defect of fusion of CD81-/- oocytes does not result from an impaired initial gamete interaction. PMID- 16380110 TI - Specification of neural precursor identity in the geophilomorph centipede Strigamia maritima. AB - Despite differences in the formation of neural precursors, all arthropod species analyzed so far generate about 30 single precursors (insects/crustaceans) or precursor groups (chelicerates/myriapods) per hemi-segment. In Drosophila, each precursor has a distinct identity conferred by segment polarity and dorso-ventral patterning genes that subdivide the ventral neuroectoderm into a grid-like structure. Temporal patterning mechanisms generate additional diversity after delamination from the neuroectoderm. Previous work shows that the genetic network involved in recruitment and specification of neural precursors is conserved in arthropods. However, comparative studies on generation of precursor diversity are few and partial. Here, we test whether aspects of the Drosophila model may apply in the geophilomorph centipede Strigamia maritima. We describe precursor formation, based on morphology and on Delta and Notch expression. We then show that in S. maritima, hunchback and Kruppel are expressed in subsets of neural precursors generating distinct temporal expression domains within the plane of the neuroectoderm. This expression pattern suggests that temporal changes in spatial patterning cues may result in the ordered production of different neural identities. We suggest that temporal patterning mechanisms were present in the last common ancestor of arthropods, although the regulatory interactions of transcription factors might have diverged in the lineage leading to insects. PMID- 16380111 TI - The adaptor protein X11Lalpha/Dmint1 interacts with the PDZ-binding domain of the cell recognition protein Rst in Drosophila. AB - The Drosophila cell adhesion molecule Rst plays key roles during the development of the embryonic musculature, spacing of ommatidia in the compound eye and of sensory organs on the antenna, as well as in the neuronal wiring of the optic lobe. In rst(CT) mutants lacking the cytoplasmic domain of the Rst protein, cell sorting and apoptosis in the eye are affected, suggesting a requirement of this domain for Rst function. To identify potential interacting proteins, yeast two hybrid screens were performed using the cytoplasmic domains of Rst and its paralogue Kirre as baits. Among several putative interactors, two paralogous Drosophila PDZ motif proteins related to X11/Mint were identified. X11/Mint family members in C. elegans (LIN-10) and vertebrates are believed to function as adaptor proteins and to regulate the assembly of multi-subunit complexes at the synapse, thereby linking the vesicle cycle to cell adhesion. Using genetic, cell biological, and biochemical approaches, we show that the interaction of Rst with X11Lalpha is of biological significance. The proteins interact, for example, in the context of cell sorting in the pupal retina. PMID- 16380112 TI - Effects of p,p'-dichlorodiphenyldichloroethylene on the expressions of transferrin and androgen-binding protein in rat Sertoli cells. AB - The mechanisms of reproductive malfunction of male mammals caused by 2,2-bis(4 chlorophenyl)-1,1-dichloroethylene (p,p'-DDE, hereafter DDE) remain unknown. To explore the effects of DDE on the expressions of transferrin (Tf) and androgen binding protein (ABP), we isolated Sertoli cells from healthy immature rats (18 20 days SD rats), set up Sertoli cell cultures, evaluated the toxicity, and measured the expression levels of mRNA of Tf and ABP genes by the one-step reverse transcriptase polymerase chain reaction method after cultured Sertoli cells were in vitro exposed to DDE at different concentrations for 24 h. The results showed that the number and survival rate of Sertoli cells decreased sharply with increased doses of DDE. The expression level of Tf mRNA decreased, whereas ABP mRNA increased gradually with increased DDE doses. There existed an obvious dose-effect relationship between the concentration of DDE and the expression levels of Tf mRNA and ABP mRNA. These findings suggest that DDE may inhibit the expression of Tf and up-modulate expression of ABP in cultured rat Sertoli cells. PMID- 16380113 TI - Suppression of feeding, drinking, and locomotion by a putative cannabinoid receptor 'silent antagonist'. AB - This study compared the effects of the putative cannabinoid receptor 'silent antagonist' O-2050 with the cannabinoid receptor inverse agonist SR 141716 on food and water consumption, and locomotor activity. Non-deprived male Wistar rats were habituated to the apparatus and testing procedures, then injected intraperitoneally with vehicle, O-2050 (0.03-3.0 mg/kg), or SR 141716 (3.0 mg/kg) prior to 4-h test sessions. Food consumption was significantly reduced by both drugs. Water intake and locomotor activity were significantly reduced only by O 2050. Results support the notion that cannabinoid receptor antagonists suppress feeding behaviour by blocking an endogenous cannabinoid orexigenic signal, rather than by inverse agonism at cannabinoid receptors. However, further studies are needed to confirm the status of O-2050 as a cannabinoid CB(1) receptor antagonist devoid of inverse agonist properties. PMID- 16380114 TI - Relaxation and cyclic GMP levels in response to sildenafil in human pulmonary arteries from donors. AB - We measured cyclic GMP formation and relaxation response to sildenafil given either alone or in combination with sodium nitroprusside (SNP) in pulmonary arteries obtained from 13 multi-organ donors. Sildenafil (10(-9)-10(-4) M) caused concentration-dependent relaxations and amplified the relaxation induced by SNP. Relaxation was unaffected by endothelium removal or by pre-treatment with the inhibitor of nitric oxide synthase L-NMMA (10(-4) M). SNP (10(-7) M) caused elevation of cyclic GMP levels that was potentiated by sildenafil (10(-6) M). Thus, the enhancement of SNP-induced relaxation by sildenafil is mainly due to an increase in cyclic GMP accumulation. PMID- 16380115 TI - Dexamethasone modifies the susceptibility to serum cytotoxicity and increases the metastatic efficiency of a colon carcinoma cell line. AB - Metastatic inefficiency is a phenomenon by which a majority of tumor cells is lost in the blood stream during the metastatic process. We investigated the effects of dexamethasone (DEX), a synthesized glucocorticoid, on the serum susceptibility of a colon carcinoma cell line, HT-29, with respect to metastatic inefficiency. The susceptibility to serum cytotoxicity of these carcinoma cells is possibly an important factor in metastatic inefficiency. In this study, we used glucocorticoid because it modifies the function of the plasma membrane and has been shown to enhance the hematogenous metastasis of some tumor cells. Using HT-29 cells that had been treated with DEX in vitro, the following factors were evaluated: the metastasis of intrasplenic injected cells; in vitro and in vivo proliferation; motility; the production of matrix metalloproteinases (MMPs); and the expression of the membrane complement regulatory proteins CD46, CD55, and CD59. The number of viable cells in the liver after an intraportal injection of tumor cells was determined by the expression of human beta-globin mRNA that is aberrantly expressed in HT-29 cells. In addition, we investigated 100% serum induced proliferation, susceptibility, and apoptosis. Treatment with DEX was found to accelerate liver metastasis; here, the number of metastatic colonies and the weight of the liver were both significantly increased in DEX-treated HT-29 (HT-29DEX) cells. In contrast, there was no difference in terms of cell motility; the production of MMPs; or the expression of CD46, CD55, or CD59 between the HT 29 and HT-29DEX cells. The HT-29DEX cells exhibited enhanced proliferation in the serum, as well as resistance to cytotoxicity when exposed to 100% serum. In addition, DEX slightly inhibited serum-induced apoptosis. Finally, the expression of colon cancer-derived beta-globin mRNA was detectable 24 h after intravenous injection, but only in samples obtained from the HT-29DEX-, but not in those from the HT-29-inoculated mice. These results indicate that DEX reduced the metastatic inefficiency of the HT-29 cells, resulting in a hematogenous metastasis-prone phenotype. It is thus expected that the acquisition of resistance against serum cytotoxicity is among the mechanisms that contribute to the efficiency of hematogeneous metastasis. PMID- 16380116 TI - Annexin A5 inhibits engulfment through internalization of PS-expressing cell membrane patches. AB - Apoptosis and subsequent clearance of apoptotic cells are important for the prevention of diseases. Therefore, it is essential to understand the mechanisms underlying the biology of phagocytic clearance of apoptotic cells. The best characterized "eat me" signal on the surface of apoptotic cells is phosphatidylserine (PS). Recently, we demonstrated that annexin A5 mediates the internalization of PS-expressing membrane patches and down regulates surface expression of tissue factor. Here, we investigated the role of PS in the phagocytosis of apoptotic cells using annexin A5. Using a novel flow cytometric based phagocytosis assay, we observed that engulfment was inhibited with 20% if annexin A5 was added to PS-expressing cells that had completed apoptosis. The inhibition increased to more than 50% if annexin A5 was added during the apoptotic process. This inhibition is specific for annexin A5, since the mutant M23 and annexin A1 did not further increase the inhibition of phagocytosis when added during the apoptotic process. Interestingly, cells with internalized annexin A5 still express PS at their surface. We conclude that other ligands within the PS-expressing membrane patch act together with PS as an "eat me" signal. PMID- 16380117 TI - Central retinal thickness is positively correlated with macular pigment optical density. AB - Macular pigment (MP) has been suggested to have a protective role in age-related macular degeneration by reducing the amount of oxidative stress on the retina. MP levels peak at the foveal center, where it is found predominantly in the receptor axon and inner plexiform layers of the retina. The purpose of this study was to investigate the relationship between central retinal thickness and macular pigment optical density in a group of healthy subjects. We report that macular pigment optical density (MPOD) has a significant and positive relationship with central retinal thickness as measured by optical coherence tomography. The strength of the observed relationship (r approximately 0.30) was independent of the technique used to measure MPOD, whether heterochromatic flicker photometry (HFP) or 2-wavelength autofluorescence (AF). Of note, there was no statistically demonstrable relationship between MPOD at an eccentricity of 1- or 2-degrees and central retinal thickness. This finding has important implications for future studies investigating MPOD, and its response to dietary modification/supplementation. PMID- 16380118 TI - Small N-terminal mutant huntingtin fragments, but not wild type, are mainly present in monomeric form: Implications for pathogenesis. AB - N-terminal fragments of huntingtin containing an expanded polyglutamine stretch play an important role in the molecular pathogenesis of Huntington's disease. Their ultimate accumulation in insoluble protein aggregates constitutes an important pathological hallmark of Huntington's disease. We report on systematic biochemical comparison studies of soluble wild type and mutant N-terminal huntingtin fragments. The results show that soluble wild type exon 1 fragments are predominantly present in higher molecular weight complexes with a molecular size of approximately 300 kDa, while their mutant counterparts are mainly present in their monomeric form. In contrast, longer N-terminal fragments corresponding to peptides produced by caspase cleavage do not display these differential properties. These findings suggest that especially an increased amount of monomeric form of small N-terminal mutant huntingtin fragments may facilitate aberrant interactions both with itself via the polyglutamine stretch and with other proteins and thereby contribute to molecular pathogenesis. PMID- 16380119 TI - Mammalian mitochondrial nitric oxide synthase: characterization of a novel candidate. AB - Recently a novel family of putative nitric oxide synthases, with AtNOS1, the plant member implicated in NO production, has been described. Here we present experimental evidence that a mammalian ortholog of AtNOS1 protein functions in the cellular context of mitochondria. The expression data suggest that a candidate for mammalian mitochondrial nitric oxide synthase contributes to multiple physiological processes during embryogenesis, which may include roles in liver haematopoesis and bone development. PMID- 16380120 TI - The translocation mechanism of P-glycoprotein. AB - Multidrug transporters are involved in mediating the failure of chemotherapy in treating several serious diseases. The archetypal multidrug transporter P glycoprotein (P-gp) confers resistance to a large number of chemically and functionally unrelated anti-cancer drugs by mediating efflux from cancer cells. The ability to efflux such a large number of drugs remains a biological enigma and the lack of mechanistic understanding of the translocation pathway used by P gp prevents rational design of compounds to inhibit its function. The translocation pathway is critically dependent on ATP hydrolysis and drug interaction with P-gp is possible at one of a multitude of allosterically linked binding sites. However, aspects such as coupling stoichiometry, molecular properties of binding sites and the nature of conformational changes remain unresolved or the centre of considerable controversy. The present review attempts to utilise the available data to generate a detailed sequence of events in the translocation pathway for this dexterous protein. PMID- 16380121 TI - Insulin regulation of glucokinase gene expression: evidence against a role for sterol regulatory element binding protein 1 in primary hepatocytes. AB - Liver key genes for carbohydrate and lipid homeostasis are regulated by insulin and glucose. The sterol regulatory-element binding protein-1c (SREBP-1c) has emerged as a mediator of insulin effects on gene transcription, particularly on glucokinase (GK). In this paper, we show that despite stimulation of GK promoter transcription by overexpression of mature SREBP-1c, insulin induced GK transcription at least 4h ahead of accumulation of mature SREBP-1c in the nucleus. Importantly, the knockdown of SREBP-1 mRNA using a RNA-interference technique reduced the level of nuclear SREBP-1 protein, diminished fatty acid synthase mRNA level, but did not affect GK and L-pyruvate kinase mRNA levels. We concluded that SREBP-1 is unlikely to be the mediator of the early insulin effect on GK gene transcription. PMID- 16380122 TI - Hirsutenone inhibits phorbol ester-induced upregulation of COX-2 and MMP-9 in cultured human mammary epithelial cells: NF-kappaB as a potential molecular target. AB - Inappropriate upregulation of cyclooxygenase-2 (COX-2) and matrix metalloproteinases (MMPs) has been implicated in the pathogenesis of various types of cancer. In the present study, we investigated the effects of hirsutenone, a diarylheptanoid isolated from the medicinal plant Alnus hirsuta var. sibirica, on the expression of COX-2 and MMP-9 induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) in MCF10A human breast epithelial cells. Treatment of MCF10A cells with TPA led to the expression of COX-2 and MMP 9. Hirsutenone at 12 microM inhibited the TPA-induced COX-2 expression at both the transcriptional and posttranscriptional levels. Hirsutenone also suppressed the synthesis of prostaglandin E(2), one of the major products of COX-2, and its catalytic activity. The upregulation of MMP-9 by TPA was also significantly reduced by hirsutenone. Likewise, hirsutenone attenuated the invasiveness and motility of MCF10A cells stimulated with TPA. Hirsutenone blocked the TPA-induced DNA binding of nuclear factor kappa B (NF-kappaB) and translocation of p65, the functionally active NF-kappaB subunit, to the nucleus. The luciferase reporter gene assay revealed that hirsutenone abrogated the transcriptional activity of NF kappaB. Treatment of MCF10A cells with N-alpha-Tosyl-l-phenylalanine chloromethyl ketone, a specific inhibitor of NF-kappaB, reduced the TPA-induced expression of COX-2 and MMP-9. In summary, hirsutenone inhibits the TPA-induced upregulation of COX-2 and MMP-9 in human breast epithelial cells, possibly by targeting NF kappaB, which may contribute to its chemopreventive effects. PMID- 16380123 TI - The hormonal and behavioral response to group formation, seasonal changes, and restraint stress in the highly social Malayan Flying Fox (Pteropus vampyrus) and the less social Little Golden-mantled Flying Fox (Pteropus pumilus) (Chiroptera: Pteropodidae). AB - This study examined behavioral and physiological responses (changes in inter animal spacing, total glucocorticoids, testosterone, and body mass) to the formation of breeding and same-sex groups in two bat species, the socially gregarious Malayan Flying Fox (Pteropus vampyrus) and the less social Little Golden-mantled Flying Fox (Pteropus pumilus). We hypothesized that social instability, especially in the breeding groups and especially in P. vampyrus, would result in elevated glucocorticoids and that social facilitation of breeding and/or male-male competition would result in persistently higher levels of testosterone in breeding males. Seasonal rhythms in all measures were also predicted, and the glucocorticoid stress response was expected to vary by sex, season, and group type. Nearly all animals responded to group formation with elevated glucocorticoids, but, for breeding animals (especially aggressive male P. vampyrus), these responses persisted over time. In both species, breeding group formation resulted in elevated testosterone in males. Glucocorticoids, testosterone, testes volume, and body mass generally peaked in the breeding season in males (late summer and early autumn), but the seasonal glucocorticoid peak in females occurred in late winter and early spring. All animals responded to restraint stress with elevations in glucocorticoids that largely did not differ by sex, time of year, reproductive condition, group type, or, in lactating females, the presence of her pup. Changes in both behavior and physiology were more evident in P. vampyrus than in P. pumilus, and we believe that their underlying social differences influenced their responses to group formation and to the changing seasonal environment. PMID- 16380124 TI - Professional roles and communications in clinical placements: a qualitative study of nursing students' perceptions and some models for practice. AB - BACKGROUND: Research evidence suggests that clinical placements are important to both the perceptions and outcomes of nurse education. Nevertheless, despite this knowledge, research also indicates that many students receive negative experiences whilst attending these 'remote' settings, sometimes resulting in missed opportunities for learning and negative impressions of potential places of employment. OBJECTIVE: In this context, this study investigates the experiences and perceptions of students relating to their clinical placements and, in particular, their views on professional structuring. Specifically it addresses the roles of, and communications between, the key academics, clinical professionals and institutions responsible for their organization. METHODS: Focus group discussions with students (n=7) and an interview survey of ex-students (n=30) each from two British universities in Southeast England. FINDINGS: The research highlights the diverse experiences that student receive on clinical placements, as well as their own suggestions for improvements. Student informed models of worst, minimum, current and best practice are then presented, as well as a cross-setting evaluation feedback model. These highlight responsibilities and communications across health professionals and educational sites. The models include an emphasis on the roles of ward managers, mentors, link tutors and more generally on shared but clearly delineated institutional responsibility for quality assurance mechanisms. They offer the opportunity to improve educational practice in clinically-based education and concurrently to improve student experiences and outcomes. CONCLUSIONS: Clinical placements are designed to provide practical learning through a 'slice of practice life'. However, it is necessary to maximize this learning experience. Placements certainly need not be the worst slice. PMID- 16380125 TI - The effect of octyl glucoside on rheological behavior of diluted and concentrated lamellar phases. AB - We have investigated the rheological properties of lamellar liquid crystal formed by nonionic surfactants at low and high surfactant concentrations with a small amount of octyl glucoside and their relationship with the topology of the bilayer. Rheology is a specific signature of each bilayer topology. The decrease in viscosity by increasing the shear rate according to a power law with exponent close to -0.8 was found for the concentrated system of LSB/1-butanol/water and 1% in weight of OG. On the contrary, the decrease in the viscosity by increasing the shear rate for the diluted system is less pronounced with lower exponent values. The rheological data agrees with the presence of vesicles. A special case is the system with benzyl alcohol. The apparent viscosity does not follow the same power law than for alkanols. PMID- 16380126 TI - Effect of temperature on the surface phase behavior of n-hexadecyl dihydrogen phosphate in adsorption layers at the air-water interface. AB - We present the adsorption kinetics and the surface phase behavior of n-hexadecyl dihydrogen phosphate (n-HDP) at the air-water interface by film balance and Brewster angle microscopy (BAM). A phase diagram, which shows a triple point at about 25.8 degrees C, is constructed by measuring the surface pressure (pi)-time (t) adsorption isotherms. Below 25.8 degrees C, each of the pi-t curves shows a plateau at about zero surface pressure indicating the existence of a first-order phase transition. The BAM observation confirms the order of this phase transition by presenting two-surface phases during this plateau. However, the BAM observation also shows clearly another second-order phase transition from an isotropic phase to a mosaic-textured liquid condensed (LC) phase. The initial phase is a gas (G) phase. Considering the peculiarity of the middle phase, we suggest this phase as an intermediate (I) phase. Above the triple point, the pi-t curves predict the existence of two-step first-order phase transitions. Similar to the results at lower temperatures, the BAM images show two-surface phases during these first-order phase transitions together with a second-order phase transition from an isotropic phase to an LC phase. These transitions are classified as a first-order G-LE (liquid expanded) phase transition, which is followed by another first-order LE-I phase transition. The second-order phase transition is an I-LC phase transition. Contrary to these results, at 36 degrees C both the pi-t measurements and the BAM observation present only two first-order phase transitions, which are G-LE at zero surface pressure and LE-LC transition at higher surface pressure. The shape of the domains during the main transitions shows a peculiar change from a circular at 20 degrees C to an elongated at 24 degrees C and finally to a circular shape at 36 degrees C. Such a change in the domain shapes has been explained considering the dehydration effect at higher temperatures as well as the nature of phases. PMID- 16380127 TI - A small-angle neutron scattering study of sodium dodecyl sulfate-poly(propylene oxide) methacrylate mixed micelles. AB - Mixed micelle of protonated or deuterated sodium dodecyl sulfate (SDS and SDSd25, respectively) and poly(propylene oxide) methacrylate (PPOMA) are studied by small angle neutron scattering (SANS). In all the cases the scattering curves exhibit a peak whose position changes with the composition of the system. The main parameters which characterize mixed micelles, i.e., aggregation numbers of SDS and PPOMA, geometrical dimensions of the micelles and degree of ionisation are evaluated from the analysis of the SANS curves. The position q(max) of the correlation peak can be related to the average aggregation numbers of SDS-PPOMA and SDSd25-PPOMA mixed micelles. It is found that the aggregation number of SDS decreases upon increasing the weight ratio PPOMA/SDS (or SDSd25). The isotopic combination, which uses the "contrast effect" between the two micellar systems, has allowed us to determine the mixed micelle composition. Finally, the SANS curves were adjusted using the RMSA for the structure factor S(q) of charged spherical particles and the form factor P(q) of spherical core-shell particle. This analysis confirms the particular core-shell structure of the SDS-PPOMA mixed micelle, i.e., a SDS "core" micelle surrounded by the shell formed by PPOMA macromonomers. The structural parameters of mixed micelles obtained from the analysis of the SANS data are in good agreement with those determined previously by conductimetry and fluorescence studies. PMID- 16380128 TI - Screening of copy number polymorphisms in human beta-defensin genes using modified real-time quantitative PCR. AB - Defensins are cationic antimicrobial peptides, which play an important role in host immune defense to some infectious diseases as well as immune disease and skin disease. Recent studies identified that the genes coding for human beta defensin 2 (DEFB4), human beta-defensin 3 (DEFB103) and human beta-defensin 4 (DEFB104) showed variation in copy numbers. This variation may have an impact on gene expression levels. Here, we have demonstrated a real-time PCR-based method to measure beta-defensin gene copy number. Using this relative real-time quantitative PCR, we developed a new rapid and reliable approach, which involves amplification of the target locus (DEFB4 or DEFB103 or DEFB104) and the single copy reference locus (human serum albumin, ALB) in a single PCR reaction. A calibrator was prepared by recombining one copy of the target gene and one copy of the reference gene into a plasmid. After correcting the PCR amplification efficiency, which differed between the defensin gene and ALB gene, and normalization by the calibrator, the ratio of the copy number of the target gene to that of the reference gene in an unknown sample was determined. This normalized ratio directly related to the gene copy number. The assay was validated using previously genotyped samples, which demonstrated high accuracy and reliability of the method. Furthermore, this method was used to screen the copy number variations of these three beta-defensin genes in healthy blood donors. This method proved to be a reliable and fast tool to genotype gene copy number variations in projects associating genomic variations with gene expression or with population phenotypes in epidemiologic studies. PMID- 16380129 TI - Alanine substitution and deletion analogues of Manduca sexta allatostatin: structure-activity relationship on the spontaneous contractions of the foregut of larval Lacanobia oleracea. AB - Manduca sexta allatostatin (Manse-AS) is a 15-residue non-amidated peptide with a blocked N-terminus and a disulphide bridge between the cysteine residues at positions 7 and 14. Analogues of Manse-AS were used to examine the structural requirements of Manse-AS for inhibitory activity on spontaneous foregut contractions of larval tomato moth (Lacanobia oleracea). Breaking the disulphide bond between C(7) and C(14) by reduction reduced the potency of the peptide, suggesting that the conformation of Manse-AS is important for its biological activity. When either of the cysteine residues were replaced with alanine the Manse-AS analogue had no measurable bioactivity. Alanine substitution at Q(6) was as potent as Manse-AS, all other alanine substitution analogues (R(5), Y(8), F(9), N(10), P(11), I(12) and S(13)), were myoinhibitory but less potent than native Manse-AS to varying degrees. Analogues with alanine substitution at amino acids with aromatic side chains (Y(8) and F(9)) were the least active. Amino terminal deletion analogues Manse-AS(6-15) and Manse-AS(7-15) were inactive whereas Manse-AS(5-15) was fully active but not as potent as Manse-AS. The results show that amino acid residues both inside and outside the disulphide ring are important for biological activity. PMID- 16380130 TI - Alanine scanning of MS2 coat protein reveals protein-phosphate contacts involved in thermodynamic hot spots. AB - The co-crystal structure of the MS2 coat protein dimer with its RNA operator reveals eight amino acid side-chains contacting seven of the RNA phosphates. These eight amino acids and five nearby control positions were individually changed to an alanine residue and the binding affinities of the mutant proteins to the RNA were determined. In general, the data agreed well with the crystal structure and previous RNA modification data. Interestingly, amino acid residues that are energetically most important for complex formation cluster in the middle of the RNA binding interface, forming thermodynamic hot spots, and are surrounded by energetically less relevant amino acids. In order to evaluate whether or not a given alanine mutation causes a global change in the RNA-protein interface, the affinities of the mutant proteins to RNAs containing one of 14 backbone modifications spanning the entire interface were determined. In three of six protein mutations tested, thermodynamic coupling between the site of the mutation and RNA groups that can be even more than 16 A away was detected. This suggests that, in some cases, the mutation may subtly alter the entire protein-RNA interface. PMID- 16380131 TI - Communication between eukaryotic translation initiation factors 5 and 1A within the ribosomal pre-initiation complex plays a role in start site selection. AB - During eukaryotic translation initiation, the 43 S ribosomal pre-initiation complex scans the mRNA in search of an AUG codon at which to begin translation. Start codon recognition halts scanning and triggers a number of events that commit the complex to beginning translation at that point on the mRNA. Previous studies in vitro and in vivo have indicated that eukaryotic initiation factors (eIFs) 1, 2 and 5 play key roles in these events. In addition, it was reported recently that the C-terminal domain of eIF1A is involved in maintaining the fidelity of start codon recognition. The molecular mechanisms by which these factors work together to ensure fidelity of start site selection remain poorly understood. Here, we report the quantitative characterization of energetic interactions between eIF1A, eIF5 and the AUG codon in an in vitro reconstituted yeast translation initiation system. Our results show that recognition of an AUG codon by the 43 S complex triggers an interaction between eIF5 and eIF1A, resulting in a shift in the equilibrium between two states of the pre-initiation complex. This AUG-dependent change may be a reorganization from a scanning competent state to a scanning-incompetent state. Mutations in both eIF1A and eIF5 that increase initiation at non-AUG codons in vivo weaken the interaction between the two factors upon AUG recognition, while specifically strengthening it in response to a UUG codon. These data suggest strongly that the interaction between eIF1A and eIF5 is involved in maintaining the fidelity of start codon recognition in vivo. PMID- 16380132 TI - Leber's hereditary optic neuropathy with dystonia in a Japanese family. AB - We investigated a Japanese family with generalized dystonia attributed to striatal degeneration, which occurred in childhood, and late-onset optic neuropathy. We determined the entire nucleotide sequence of mitochondrial DNA (mtDNA) from the proband and compared our findings with the 2001 Revised Cambridge Reference Sequence. The mtDNA of the proband showed a total of 42 nucleotide changes. We identified two A3203G and G14459A mutations, which were completely absent in a population of 200 healthy Japanese, by estimating the frequency of each nucleotide change. The nucleotide G14459A mutation occurs in NADH dehydrogenase subunit 6, and has been suggested previously as the disease causing mutation in Hispanic, African-American and Caucasian families of Leber's hereditary optic neuropathy (LHON) and/or dystonia. The significance of the A3203G mutation remains unknown. To our knowledge, this is the first case of LHON with dystonia that revealed a mtDNA mutation in a Japanese family. PMID- 16380134 TI - Clinicopathologic features of nonsystemic vasculitic neuropathy and microscopic polyangiitis-associated neuropathy: a comparative study. AB - OBJECTIVE: To compare clinicopathologic findings in nonsystemic vasculitic neuropathy (NSVN) and microscopic polyangiitis-associated neuropathy (MPAN). METHODS: Patients clinicopathologically confirmed to have NSVN (n=23) or MPAN (n=40) were compared with respect to clinical, electrophysiologic, and histopathologic features. RESULTS: Clinical features of neuropathy such as initial symptoms, progression, and distribution of sensory and motor deficits were similar in both groups, while functional compromise was greater in MPAN than NSVN. Abnormalities of laboratory data including those reflecting severity and extent of inflammation such as C-reactive protein were more conspicuous in MPAN than NSVN. Perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCA) were positive in two-thirds of patients with MPAN but negative in all NSVN. Electrophysiologic and histopathologic findings indicated axonal neuropathy in both groups, whereas the reduction of compound muscle action potentials in the tibial nerve and sensory nerve action potentials in the median nerve was significantly more profound in MPAN than NSVN. As for the epineurial perivascular infiltration, frequencies of cell-specific markers for T lymphocytes, macrophages, and B lymphocytes among cells infiltrating the vasculitic lesions were essentially similar between groups. CONCLUSIONS: Clinicopathologic profiles and vascular pathology were similar between NSVN and MPAN but the age at onset, severity, and presence of p-ANCA were clearly different. Further study is needed to clarify the pathogenesis of NSVN and its place in the vasculitic spectrum of diseases. PMID- 16380133 TI - Mutation analysis of the ATM gene in two Chinese patients with ataxia telangiectasia. AB - Ataxia telangiectasia (A-T) is an autosomal recessive disorder characterized by cerebellar ataxia, telangiectasia, immunodeficiency, elevated alpha-fetoprotein level, chromosomal instability, predisposition to cancer, and radiation sensitivity. Although a lot of mutations in the ATM gene have been described, there is still no report about ATM mutations in Chinese population. Using a molecular approach, we screened for ATM mutations in two patients from two unrelated Chinese families. 100 normal controls were analyzed to exclude possibility of polymorphism. Two novel mutations in the ATM gene were identified. The first one is a novel, homozygous, 1346G>C (Gly449Ala) missense mutation. The second one is a compound heterozygous mutation, which consists of a novel, 610G>T (Gly204Stop) nonsense mutation, combined with a previously reported, 6679C>T (Arg2227Cys) missense mutation. The transversions 1346G>C (Gly449Ala) and 610G>T (Gly204Stop) are not localized either in the conserved PI-3 kinase domain or in the other domains of the ATM protein. The phenotypic features were characterized by progressive cerebellar ataxia, ocular telangiectasia, elevated alpha fetoprotein level, immunodeficiency (agammaglo-bulinemia and T-cell defect), and rearrangements of chromosomes 7 and 14; brain MRI showed cerebellar atrophy, brain SPECT showed cerebellar regional cerebral blood flow (rCBF) hypoperfusion. To our knowledge, this is the first report of ATM mutations in Mainland China, in which the transversions 1346G>C (Gly449Ala) and 610G>T (Gly204Stop) are two novel, disease-causing mutations. PMID- 16380136 TI - A longitudinal survey of self-assessed health trends in a community cohort of people with multiple sclerosis and their significant others. AB - BACKGROUND: Studies assessing psychosocial consequences of multiple sclerosis (MS) in the community are scarce; it appears that there are no longitudinal surveys in this area. OBJECTIVES: We prospectively assessed changes in self perceived health status over 5 years in a community cohort of MS adults. METHODS: The 251 people who participated in a 1999 postal survey were re-assessed in 2004, being sent the Multiple Sclerosis Quality-of-Life-54 (MSQOL-54), the Chicago Multiscale Depression Inventory (CMDI), and a demographic/clinical questionnaire. Health-related quality of life (Short Form-36) and CMDI were also assessed in participants' significant others. RESULTS: A total of 205 people participated: 14 (5.6%) of the original cohort MS had died and 32 (13%) did not return the questionnaires. A significant other was available for 74% of responders. The proportion requiring constant bilateral walking assistance increased from 16% to 33%. The proportion using housing adaptations increased from 17% to 27%, and the use of daily home care increased from 19% to 28%. Impaired CMDI mood affected 27% of MS and 19% of significant others. Changes in MSQOL-54 were not unidirectional: the domains change in health, physical function, and general health worsened; while social function, mental health, and health distress improved significantly. CONCLUSIONS: MS has a pervasive but inhomogeneous impact on the lives of MS sufferers: the proportion of those severely impaired doubled over the study period; nevertheless in 23% of participants the disease remained mild over a median duration of 11 years. The psychological burden affects not only people with MS but also their significant others. PMID- 16380137 TI - Fitting of random tessellation models to keratin filament networks. AB - The role of specific structural patterns in keratin filament networks for regulating biophysical properties of epithelial cells is poorly understood. This is at least partially due to a lack of methods for the analysis of filament network morphology. We have previously developed a statistical approach to the analysis of keratin filament networks imaged by scanning electron microscopy. The segmentation of images in this study resulted in graph structures, i.e. tessellations, whose structural characteristics are now further investigated by iteratively fitting geometrical statistical models. An optimal model as well as corresponding optimal parameters are detected from a given set of possible random tessellation models, i.e. Poisson-Line tessellations (PLT), Poisson-Voronoi tessellations (PVT) and Poisson-Delaunay tessellations (PDT). Using this method, we investigated the remodeling of keratin filament networks in pancreatic cancer cells in response to transforming growth factor alpha (TGFalpha), which is involved in pancreatic cancer progression. The results indicate that the fitting of random tessellation models represents a suitable method for the description of complex filament networks. PMID- 16380138 TI - Successive alterations of hippocampal gamma-aminobutyric acid B receptor subunits in a rat model of febrile seizure. AB - Febrile seizure (FS) is a frequently encountered seizure type in childhood. Changes of brain function following FS have clinical importance. The recently identified gamma-aminobutyric acid B receptor (GABA(B)R) is a metabotropic receptor of GABA. In this study, we used a rat model of recurrent FS to investigate the changes of GABA(B)R1a and GABA(B)R2 subunits in hippocampus after recurrent FS by using Western blot, quantitative RT-PCR, double immunofluorescence, in situ hybridization and immunoprecipitation/Western blot. After treatment of hyperthermia and the presence of induced seizures once every 2 days for 10 times, GABA(B)R1a and GABA(B)R2 subunits in hippocampus were decreased after 24 h of the last treatment. The decrease of GABA(B)R1a lasted for 15 days but that of GABA(B)R2 persisted for more than 30 days. The binding of GABA(B)R1a to GABA(B)R2 in hippocampus was also decreased significantly after 24 h of the last treatment and lasted for more than 30 days. In situ hybridization showed that GABA(B)R1a mRNA was significantly decreased in dentate gyrus, and GABA(B)R2 mRNA was considerably reduced in CA3 region. In H10 and FS1 groups in which hyperthermia treatment was the same but no (H10 group) or only one seizure (FS(1) group) was induced, the decrease of GABA(B)R1a and GABA(B)R2 subunits and the reduced binding capability between GABA(B)R1a and GABA(B)R2 subunits were also detected but with less severity, and the time recovering from these abnormalities was shorter. We conclude that GABA(B)R1a and GABA(B)R2 subunits and the binding of the 2 subunits decrease in hippocampus for a relatively long period of time after recurrent FS in immature rats. These changes may result in long-lasting imbalance of excitation/inhibition function in hippocampus, and are derived from the consequences of recurrent febrile seizures. PMID- 16380139 TI - Biomarker responses as indication of contaminant effects in blue mussel (Mytilus edulis) and female eelpout (Zoarces viviparus) from the southwestern Baltic Sea. AB - During a field study performed in spring and autumn 2001 and 2002, blue mussels (Mytilus edulis) and female eelpout (Zoarces viviparus) were collected at three locations in the Wismar Bay (Baltic Sea), and several biomarkers of contaminant effects were analysed. Besides seasonal and inter-annual variations, biomarker signals were most pronounced at the location closest to Wismar Harbour (Wendorf) in both species. Lysosomal membrane stability (LMS) was lowest and acetylcholinesterase activity (AChE) was significantly reduced. Frequency of micronuclei (MN) was significantly higher (in blue mussels), indicating mutagenic effects. In eelpout elevated levels of DNA adducts, EROD induction and PAH metabolites were measured. Metallothionein (MT), biomarker for trace metal exposure, showed a gradient only in spring. Organochlorine contaminant analyses (PCBs, DDTs) corresponded to the observed biomarker levels. The results obtained clearly demonstrate pollution effects in the southwestern Baltic Sea. Moreover, they show that a multibiomarker approach is also applicable in a brackish water environment. PMID- 16380140 TI - Biomarkers of exposure and effect in the Brazilian flounder Paralichthys orbignyanus (Teleostei: Paralichthyidae) from the Patos Lagoon estuary (Southern Brazil). AB - Biomarkers of exposure (liver metallothionein-like proteins content and catalase and glutathione S-transferase activities) and effect (liver lipoperoxidation and blood cell DNA damage) of contaminants were analyzed in the Brazilian flounder Paralichthys orbignyanus from the Patos Lagoon estuary (Southern Brazil). Flounders were collected for a year in two sites: "Coroa do Boi" (polluted site) and "Saco do Justino" (non-polluted site). Results indicated that micronucleated cells frequency was the best biomarker to distinguish flounders from the two sites. Taken together, data from DNA damage analyses (micronucleus test and comet assay) indicated that flounders from the non-polluted site efficiently repaired the DNA breaks, contrary to those from the polluted site, which probably had their DNA repair system inhibited or exhausted. Furthermore, data from enzyme activities (catalase and GST) and lipid peroxidation indicated that flounders from the polluted site were under oxidative stress in summer and autumn. PMID- 16380141 TI - Cytogenetic and cytotoxic effects in gill cells of the blue mussel Mytilus spp. from different zones of the Baltic Sea. AB - Frequency of micronuclei (MN) and other nuclear abnormalities (nuclear buds, bi nucleated and fragmented-apoptotic cells) was analysed in gill cells of the blue mussel (Mytilus spp.) from selected coastal sites in the Baltic Sea--Kvadofjarden (Sweden), Klaipeda-Butinge zone (Lithuania), Gulf of Gdansk (Poland) and Wismar Bay (Germany). Samples were collected from 650 specimens during bi-annual sampling campaigns in 2001 and 2002. The lowest frequency of MN (0.37 MN/1000 cells) was found in blue mussels from the reference site (Kvadofjarden). The highest MN values (up to 6.7 MN/1000 cells) were registered in blue mussels from the Gulf of Gdansk in autumn 2001 and 2002, and at Wismar Bay in spring 2001 (up to 5.06 MN/1000 cells). Gradients of MN incidences were observed when comparing the three studied locations in Wismar Bay, and at the Lithuanian coast before the crude oil spill in the Butinge oil terminal. Moreover, significant seasonal and inter-location differences in the responses were documented (P<0.0001). Nuclear abnormalities were observed most frequently in blue mussels from the Gulf of Gdansk. PMID- 16380142 TI - Biomarkers in croakers Micropogonias furnieri (Teleostei: Sciaenidae) from polluted and non-polluted areas from the Patos Lagoon estuary (Southern Brazil): evidences of genotoxic and immunological effects. AB - Biomarkers of exposure and effect of pollutants were analyzed in croakers Micropogonias furnieri (Teleostei: Sciaenidae) captured in winter and summer in a polluted and in a non-polluted site at the Patos Lagoon estuary (Southern Brazil). Catalase and glutathione S-transferase activities (exposure biomarkers) and lipid peroxidation (effect biomarker) were analyzed in liver samples. Other two effect biomarkers were also studied: blood cells DNA damage (through comet assay and micronucleus test) and respiratory burst measurements. In a broad view, results point to an important seasonal variation of the biochemical biomarkers analyzed. However, data obtained clearly indicate that croakers collected in winter at the polluted site were subjected to a level of clastogenic agents sufficient to generate irreversible genetic damages (mutations) and impair the fish immune system. PMID- 16380143 TI - Heparin-binding and patterns of virulence for two recombinant strains of Sindbis virus. AB - E2 is an important determinant of Sindbis virus neurovirulence. Increased heparan sulfate (HS) binding is associated with rapid clearance of viremia and usually with decreased virulence. However, substitution of histidine for arginine at E2 157 (R157H) or glutamate for lysine at E2-159 (K159E) produces viruses with decreases in heparin-Sepharose binding and increases in viremia but different levels of binding to HS-expressing cells and virulence phenotypes in newborn CD-1 mice (Byrnes, A.P., Griffin, D.E., 2000. Large-plaque mutants of Sindbis virus show reduced binding to heparan sulfate, heightened viremia and slower clearance from the circulation. J. Virol. 74, 644-651). To identify mechanisms of virulence, R157H and K159E were studied in newborn CD-1 and BALB/c mice. Subcutaneous inoculation of R157H caused 100% and K159E 60% mortality in 2-day old CD-1 mice. R157H caused 25% and K159E no mortality in 2-day-old BALB/c mice. R157H and K159E replicated similarly at the site of inoculation with the same level of viremia, but clearance was slower in CD-1 than BALB/c mice. R157H replicated better than K159E in the central nervous system (CNS) after subcutaneous and intracerebral inoculation and in undifferentiated neurons. These studies show a genetic restriction of replication in newborn BALB/c mice, and that amino acid substitutions affecting binding to proteoglycans may differ in importance for CNS infection and viremia. PMID- 16380144 TI - Mutations at the C-terminus of the simian immunodeficiency virus envelope glycoprotein affect gp120-gp41 stability on virions. AB - The transmembrane (TM) subunit of the envelope (Env) glycoprotein of the simian immunodeficiency virus (SIV) contains an unusually long cytoplasmic domain of 164 amino acids. Previously, we identified domains in the SIV TM cytoplasmic tail that are necessary for Env incorporation into virions and viral infectivity. In this study, we investigated the relevance to Env function of the highly conserved sequence comprising the immediate C-terminal 19 residues of TM. To this end, small in-frame deletions as well as a premature stop codon mutation were introduced into the coding region for the SIV TM C-terminus. All the mutant Env glycoproteins were expressed, processed and transported to the cell surface in an essentially wild-type manner. Moreover, the ability of the mutant Env proteins to mediate cell-to-cell fusion was similar to or slightly lower than that of the wild-type Env. However, viruses expressing the mutant Env glycoproteins were found to be poorly infectious in single-cycle infectivity assays. Further characterization of the TM mutant viruses revealed that while exhibiting wild type levels of the TM protein, they contained significantly lower levels of the Env surface (SU) subunit, which is consistent with increased SU shedding from virions after Env incorporation. This phenotype was independent of Gag processing, since genetic inactivation of the viral protease did not increase SU retention by the resulting immature particles. Our findings indicate that deletions at the C-terminus of the SIV Env promote the instability of the SU-TM association on the virion surface and point to an important role for the TM cytoplasmic domain in modulating Env structure. PMID- 16380145 TI - Diagnostic approaches for viruses and prions in stem cell banks. AB - Some stem cell lines may contain an endogenous virus or can be contaminated with exogenous viruses (even of animal origin) and may secrete viral particles or express viral antigens on their surface. Moreover, certain biotechnological products (e.g. bovine fetal serum, murine feeder cells) may contain prion particles. Viral and prion contamination of cell cultures and "feeder" cells, which is a common risk in all biotechnological products derived from the cell lines, is the most challenging and potentially serious outcome to address, due to the difficulty involved in virus and prion detection and the potential to cause serious disease in recipients of these cell products. Stem cell banks should introduce adequate quality assurance programs like the microbiological control program and can provide researchers with valuable support in the standardization and safety of procedures and protocols used for the viral and prion testing and in validation programs to assure the quality and safety of the cells. PMID- 16380146 TI - Polydnavirus genomes reflect their dual roles as mutualists and pathogens. AB - Symbionts often exhibit significant reductions in genome complexity while pathogens often exhibit increased complexity through acquisition and diversification of virulence determinants. A few organisms have evolved complex life cycles in which they interact as symbionts with one host and pathogens with another. How the predicted and opposing influences of symbiosis and pathogenesis affect genome evolution in such instances, however, is unclear. The Polydnaviridae is a family of double-stranded (ds) DNA viruses associated with parasitoid wasps that parasitize other insects. Polydnaviruses (PDVs) only replicate in wasps but infect and cause severe disease in parasitized hosts. This disease is essential for survival of the parasitoid's offspring. Thus, a true mutualism exists between PDVs and wasps as viral transmission depends on parasitoid survival and parasitoid survival depends on viral infection of the wasp's host. To investigate how life cycle and ancestry affect PDVs, we compared the genomes of Campoletis sonorensis ichnovirus (CsIV) and Microplitis demolitor bracovirus (MdBV). CsIV and MdBV have no direct common ancestor, yet their encapsidated genomes share several features including segmentation, diversification of virulence genes into families, and the absence of genes required for replication. In contrast, CsIV and MdBV share few genes expressed in parasitized hosts. We conclude that the similar organizational features of PDV genomes reflect their shared life cycle but that PDVs associated with ichneumonid and braconid wasps have likely evolved different strategies to cause disease in the wasp's host and promote parasitoid survival. PMID- 16380147 TI - Psychophysics of reading with a limited number of pixels: towards the rehabilitation of reading ability with visual prosthesis. AB - Psychophysics of reading with limited numbers of pixels has received increasing attention as the envisioned visual prosthesis offers a possibility to restore some useful reading ability to the blind through the pixelized vision it generates. This paper systematically studied how several important parameters of pixelized vision affect reading performance. A closed-circuit television reading platform with digital image processing capacities was developed to convert images of printed text into pixelized patterns made up of discrete dots. Reading rates in six normally sighted subjects were measured under different combinations of pixel number, window width, and angular subtense of pixel array. The results showed that reading is possible with as few as 6 x 6 binary pixels, at 15 words/min. It was also found that for a given array of pixels, maximum reading rates occur at a specific medium window width, due to a tradeoff between window width and character sampling resolution. It was also observed that pixelized reading exhibits more significant scale dependence than normal vision. Reading rates were decreased by increasing the angular subtense of the pixel array while keeping other parameters fixed. We hope these results will be helpful to the design of visual prosthesis for the rehabilitation of reading abilities. PMID- 16380148 TI - Equilibrium and kinetic modelling of Cd(II) biosorption by algae Gelidium and agar extraction algal waste. AB - In this study an industrial algal waste from agar extraction has been used as an inexpensive and effective biosorbent for cadmium (II) removal from aqueous solutions. This biosorbent was compared with the algae Gelidium itself, which is the raw material for agar extraction. Equilibrium data follow both Langmuir and Redlich-Peterson models. The parameters of Langmuir equilibrium model are q(max)=18.0 mgg(-1), b=0.19 mgl(-1) and q(max)=9.7 mgg(-1), b=0.16 mgl(-1), respectively for Gelidium and the algal waste. Kinetic experiments were conducted at initial Cd(II) concentrations in the range 6-91 mgl(-1). Data were fitted to pseudo-first- and second-order Lagergren models. For an initial Cd(II) concentration of 91 mgl(-1) the parameters of the pseudo-first-order Lagergren model are k(1,ads)=0.17 and 0.87 min(-1); q(eq)=16.3 and 8.7 mgg(-1), respectively, for Gelidium and algal waste. Kinetic constants vary with the initial metal concentration. The adsorptive behaviour of biosorbent particles was modelled using a batch reactor mass transfer kinetic model. The model successfully predicts Cd(II) concentration profiles and provides significant insights on the biosorbents performance. The homogeneous diffusivity, D(h), is in the range 0.5-2.2 x10(-8) and 2.1-10.4 x10(-8)cm(2)s(-1), respectively, for Gelidium and algal waste. PMID- 16380149 TI - Effect of operating parameters on molinate biodegradation. AB - The effect of operating conditions during molinate degradation by the defined mixed bacterial culture DC, previously described as able to mineralize molinate, was evaluated in a batch reactor. Parameters such as the rate of molinate degradation, the dissolved organic carbon (DOC) consumption and the accumulation of molinate degradation products were monitored along the culture growth. The effect of conditions such as temperature, pH, aeration rate, salinity, and presence of additional carbon and/or nitrogen sources, was tested independently. Degradation of molinate in river water was also evaluated. Culture DC was able to grow and to mineralize molinate at all the conditions assayed. Temperature was the factor with the strongest influence on bacterial growth and molinate mineralization. The lowest and the highest rate values of growth (0.010 and 0.110 h(-1)) and of molinate degradation (0.027 and 0.180 g molinate g(-1)celldrywth( 1)) were obtained at 15 and 35 degrees C, respectively. In cultures with approximately 187 mgl(-1) of molinate, 2-oxo-molinate was the major molinate degradation product accumulated in the medium, in concentrations below 0.133 mgl( 1). Degradation of molinate was also evaluated in a continuous stirred tank reactor (CSTR). Operating the CSTR at a hydraulic retention time (HRT) of 83 h, fed with medium containing molinate concentrations ranging from 1 to 3mM, culture DC degraded the herbicide with specific degradation rates similar to those obtained in the batch systems. PMID- 16380150 TI - Formation of nitrosodimethylamine (NDMA) during chlorine disinfection of wastewater effluents prior to use in irrigation systems. AB - The probable human carcinogen nitrosodimethylamine (NDMA) is produced when wastewater effluent is disinfected with chlorine. In systems where wastewater effluent is used for landscape or crop irrigation, relatively high chlorine doses (i.e., up to 2,000,mg-min/L) are often used to ensure adequate disinfection and to minimize biofouling in the irrigation system. To assess the formation of NDMA in such systems, samples were collected from several locations in full-scale wastewater treatment systems and their associated irrigation systems. Up to 460 ng/L of NDMA was produced in full-scale systems in which chloramines were formed when wastewater effluent was disinfected with chlorine in the presence of ammonia. Less than 20 ng/L of NDMA was produced in systems that used free chlorine (i.e., HOCl/OCl(-)) for disinfection in the absence of ammonia. The production of NDMA in ammonia-containing systems was correlated with the concentration of NDMA precursors in the wastewater effluent and the overall dose of chlorine applied. Much of the NDMA formation occurred in chlorine contact basins or in storage basins where water that contained chloramines was held after disinfection. When landscape or crop irrigation is practiced with ammonia containing wastewater effluent, NDMA production can be controlled by use of lower chlorine doses or by application of alternative disinfectants. PMID- 16380151 TI - Electrokinetic dewatering of Turkish glass sand plant tailings. AB - In this study, the dewatering of glass sand plant tailings from Mersin, Turkey was investigated using an electrokinetic technique. The particle size (d(80)) of the solid waste material tested was less than 0.020 mm and consisted mainly of silica, orthoclase, alumina, potassium and iron oxides. In current plant practice, Larox high-pressure filters are used to produce a filter cake containing 22-25% moisture. As an alternative, a laboratory-scale dewatering pond using an electrokinetic technique was designed and a final product (cake) containing 34% moisture was obtained after 24h treatment using 14.6 V direct current applied for 40 min. When the treatment continued up to 48 h, a firm-to hard cake was produced. PMID- 16380152 TI - Removal of a cationic dye from aqueous solutions by adsorption onto bentonite clay. AB - The ability of bentonite to remove malachite green from aqueous solutions has been studied for different adsorbate concentrations by varying the amount of adsorbent, temperature, pH and shaking time. Maximum adsorption of the dye, i.e. >90% has been achieved in aqueous solutions using 0.05 g of bentonite at a pH of 9. Thermodynamic parameters such as DeltaH degrees, DeltaS degrees and DeltaG degrees were calculated from the slope and intercept of the linear plots of lnK(D) against 1/T. Analysis of adsorption results obtained at 298, 308, 318 and 328 K showed that the adsorption pattern on bentonite seems to follow the Langmuir, Freundlih and D-R isotherms. The temperature increase reduces adsorption capacity by bentonite, due to the enhancement of the desorption step in the mechanism. The numerical values of sorption free energy (E(a)) of 1.00 1.12 kJ mol(-1) indicated physical adsorption. The kinetic data indicated an intraparticle diffusion process with sorption being first order. The rate constant k was 0.526 min(-1). The concentration of malachite green oxalate was measured before and after adsorption by using UV-Vis spectrophotometer. PMID- 16380153 TI - Effect of patient location on the performance of clinical models to predict pulmonary embolism. AB - INTRODUCTION: Current clinical likelihood models for predicting pulmonary embolism (PE) are used to categorize outpatients into low, intermediate and high clinical pre-test likelihood of PE. Since these clinical prediction rules were developed using outpatients it is not known if they can be applied universally to both inpatients and outpatients with suspected PE. Thus, the purpose of this study was to determine the effect of patient location on the performance of clinical models to predict PE. MATERIALS AND METHODS: Two clinical models (Wells and Wicki) were applied to data from the multi-centered PIOPED study. The Wells score was applied to 1359 patients and the Wicki score was applied to 998 patients. 361 patients (27%) from the PIOPED study did not have arterial gas measurement and were excluded from the Wicki score patient group. Patients were stratified by their location at the time of entry into the PIOPED study as follows: outpatient/emergency, surgical ward, medicine/coronary care unit or intensive care unit. The diagnostic performance of the two clinical models was applied to the various patient locations and the performance was evaluated using the area under a fitted receiver operating characteristic curve (AUC). RESULTS: The prevalence of PE in the three clinical probability categories were similar for the two scoring methods. Both clinical models yielded the lowest diagnostic performance in patients referred from surgical wards. The AUC for both clinical prediction rules decreased significantly when applied to inpatients in comparison to outpatients. CONCLUSIONS: Current clinical prediction rules for determining the pre-test likelihood of PE yielded different diagnostic performances depending upon patient location. The performance of the clinical prediction rules decreased significantly when applied to inpatients. In particular, the rules performed least well when applied to patients referred from surgical wards suggesting these rules should not be used in this patient group. As expected the clinical prediction rules performed best in outpatients with the optimum diagnostic performance in patients referred from emergency and outpatient wards. PMID- 16380154 TI - Analyzing the mechanism of Rap1 activation in platelets: Rap1 activation is related to the release reaction mediated through the collagen receptor GPVI. AB - The abundant Rap1 in platelets becomes activated when these cells are stimulated by various agonists, but its function has remained unknown. In view of this, we developed an assay to quantitatively measure activated Rap1 and used it to determine relationships between Rap1 activation and several platelet functions: integrin alpha2beta1 activation, tyrosine phosphorylation, and the release reaction. We looked at how these processes are affected by the protein kinase C inhibitor BIMI, tyrosine kinase inhibitor PP2, PI 3-kinase inhibitor wortmannin, and ADP scavenger apyrase. In CRP (collagen related peptide)-activated platelets, all the inhibitors severely inhibited Rap1 activation, but had little effect on integrin alpha2beta1 activation, indicating that the integrin activation mechanism is different from the Rap1 activation mechanism, at least in GPVI dependent activation. With p85alpha-null mouse platelets, we demonstrated that Rap1 activation involves PI 3-kinase p85alpha-dependent tyrosine phosphorylation. All the inhibitors similarly decreased Rap1 activation and the serotonin release reaction, and the inhibition of Rap1 activation was not due to the lack of released ADP. Our results indicate that platelet Rap1 activation is closely related to the release reaction and not to integrin alpha2beta1 activation in GPVI-mediated platelet activation. PMID- 16380155 TI - Cryptobiosis: a new theoretical perspective. AB - The tardigrade is a microscopic creature that under environmental stress conditions undergoes cryptobiosis [Feofilova, E.P., 2003. Deceleration of vital activity as a universal biochemical mechanism ensuring adaptation of microorganisms to stress factors: A review. Appl. Biochem. Microbiol. 39, 1-18; Nelson, D.R., 2002. Current status of the tardigrada: Evolution and ecology. Integrative Comp. Biol. 42, 652-659]-a temporary metabolic depression-which is considered to be a third state between life and death [Clegg, J.S., 2001. Cryptobiosis-a peculiar state of biological organization. Comp. Biochem. Physiol. Part B 128, 613-624]. In contrast with death, cryptobiosis is a reversible state, and as soon as environmental conditions change, the tardigrade "returns to life." Cryptobiosis in general, and among the tardigrade in particular, is a phenomenon poorly understood [Guppy, M., 2004. The biochemistry of metabolic depression: a history of perceptions. Comp. Biochem. Physiol. Part B 139, 435-442; Schill, R.O., et al., 2004. Stress gene (hsp70) sequences and quantitative expression in Milensium tardigradum (Tardigrade) during active and cryptobiotic stages. J. Exp. Biol. 207, 1607-1613; Watanabe, M., et al., 2002. Mechanisn allowing an insect to survive complete dehydration and extreme temperatures. J. Exp. Biol. 205, 2799 2802; Wright, J.C., 2001. Cryptobiosis 300 years on from van Leuwenhoek: what have we learned about tardigrades? Zool. Anz. 240, 563-582]. Moreover, the ability of the tardigrade to bootstrap itself and to return to life seems paradoxical like the legendary Baron von Munchausen who pulled himself out of the swamp by grabbing his own hair. Two theoretical obstacles prevent us from advancing our knowledge of cryptobiosis. First, we lack appropriate theoretical understanding of reversible processes of biological computation in living systems. Second, we lack appropriate theoretical understanding of bootstrapping in living systems. In this short opinion article, I would like to present the idea that although cryptobiosis is obscure from a certain point of view, it makes sense within a scientific perspective suggesting that "organization becomes cause in the matter" [Strohman, R.C., 2000. Organization becomes cause in the matter. Nat. Biotechnol. 18, 575-576]. I present Bateson's idea that organisms have a "recursive hierarchical" form of organization [Neuman, Y., 2004. Meaning making in the immune system. Perspect. Biol. Med. 48, 320-327; Neuman, Y., in press. A theory of meaning. Inform. Sci.] and suggest that this form of organization allows bootstrapping through reversible process of computation as discussed by theoretical physicists [Bennett, C.H., 1982. The thermodynamics of computation-a review. Int. J. Theoret. Phys. 1, 905-940; Landauer and Bennett, 1985]. PMID- 16380156 TI - The PTEN tumor suppressor inhibits telomerase activity in endometrial cancer cells by decreasing hTERT mRNA levels. AB - OBJECTIVES: Loss of PTEN expression is one of the most prevalent and earliest molecular abnormalities associated with endometrial carcinogenesis. Given that PTEN is often absent and telomerase is overexpressed by endometrial cancers, we hypothesize that PTEN signaling is important in telomerase regulation. METHODS: PTEN expression was reconstituted in the PTEN-null Ishikawa endometrial cancer cells by adenovirus-mediated gene transduction. Cell proliferation was evaluated 12-96 h after infection. Western blot analysis was performed to assess PTEN status and phosphorylated Akt expression. Telomerase activity was determined by the telomeric repeat amplification protocol (TRAP) assay. hTERT mRNA levels were assessed by real-time RT-PCR. Ishikawa cells were also treated with LY294002, a PI3-kinase inhibitor. RESULTS: Infection of Ishikawa cells by replication defective recombinant adenovirus expressing wild-type PTEN, but not control adenovirus or adenovirus expressing lipid phosphatase defective PTEN GE mutant, inhibited constitutive Akt activation and suppressed proliferation of Ishikawa cells. Infection by wild-type PTEN adenovirus, but not control adenovirus, inhibited telomerase activity 24 h after infection. This inhibition of telomerase activity was parallel to decreased hTERT mRNA levels. LY294002 treatment resulted in dose-dependent inhibition of Akt activation and cellular proliferation. LY294002 suppressed telomerase activity and decreased hTERT transcript levels in a dose-dependent manner. CONCLUSIONS: Our data suggest that PTEN may regulate telomerase activity by a novel mechanism in which inhibition of Akt activation by PTEN leads to decreased hTERT mRNA levels. Thus, loss of PTEN may allow endometrial cells to continue to express high levels of telomerase activity, facilitating the neoplastic transformation of the endometrium. PMID- 16380157 TI - Molecular profiling of endometrial cancers from African-American and Caucasian women. AB - OBJECTIVE: It is widely recognized that racial disparity in survival exists between African-American and Caucasian women with endometrial cancer (EC). Differential mutation frequencies in select genes have been postulated to explain these survival differences. The purpose of this study was to test the hypothesis that African-American women with EC have a distinct gene expression profile compared to Caucasian women with EC. METHODS: Microarray-based expression profiling using the Affymetrix U133A oligonucleotide array was performed on a series of ECs from African-American (n = 14) and Caucasian (n = 25). The two groups were matched for possible confounding variables including stage, histologic grade, and subtype. A model-based class comparison analysis was performed to generate a list of differentially expressed genes using a P value of <0.001. RESULTS: The class comparison analysis of genes differentially expressed by tumors from the two groups revealed 16 genes differentially expressed at P < 0.001, which was not statistically significant (P = 0.68). In addition, hierarchical clustering analysis did not segregate these tumors into two distinct groups based on race. CONCLUSION: These data indicate that there is no discernible difference in global gene expression profiles of ECs from African American and Caucasian women. Thus, racial disparities in clinical outcomes are unlikely to reflect differences in gene expression and may instead be attributable to other epidemiologic, clinical, or pathologic factors. PMID- 16380159 TI - Antidepressant-like effect of icariin and its possible mechanism in mice. AB - The behavioral, neurochemical and neuroendocrine effects of icariin isolated from Epimedium brevicornum were investigated in behavioral despair models of KunMing strain of male mice. Icariin was found to significantly shorten immobility time in the forced swimming test (FST) after orally administration for 21 consecutive days. Icarrin also produced a marked reduction in immobility time in the tail suspension test (TST) when administered for at least 7 consecutive days. The preferable antidepressant action by icariin was obtained at 17.5 and 35 mg/kg in the present study. Moreover, it was observed that the stress of FST exposure induced increases in brain monoamine oxidase (MAO) A and B activities, serum corticotropin-releasing factor (CRF) levels, as well as decreases in brain monoamine neurotransmitter levels. Treatment of icariin for 21 consecutive days mainly reversed the above effects in the mouse FST. These results suggested that icarrin possessed potent antidepressant-like properties that were mediated via neurochemical and neuroendocrine systems. PMID- 16380160 TI - Analysis of residual monomers in dendritic methacrylate copolymers and composites by HPLC and headspace-GC/MS. AB - OBJECTIVES: The aim of this study was to analyze the residual monomer content of photopolymerized dendritic methacrylate copolymers and particulate filler composites. Headspace-gas chromatography/mass spectrometry (HS-GC/MS) was compared with high performance liquid chromatography (HPLC). METHODS: The resin mixtures consisted of a dendritic methacrylate monomer, methyl methacrylate and acetoacetoxyethyl methacrylate in varied proportions. In addition, one of the composites contained 1,4-butanediol dimethacrylate. Camphorquinone and 2-(N,N dimethylamino)ethyl methacrylate were used as the light-activated initiator system. The content of residual methyl methacrylate and acetoacetoxyethyl methacrylate after 40 s photopolymerization were analyzed with HPLC and HS-GC/MS. RESULTS: The content of residual methyl methacrylate decreased and residual acetoacetoxyethyl methacrylate increased with increasing concentration of acetoacetoxyethyl methacrylate in the resin mixture. The results with both methods had the same trend. SIGNIFICANCE: The addition of acetoacetoxyethyl methacrylate enhanced the copolymerization of methyl methacrylate, but did not decrease the total residual monomer content. The HS-GC/MS method was found to be a feasible method in the analysis of low-boiling residuals in dental polymers. PMID- 16380161 TI - pH triggered release of protective poly(ethylene glycol)-b-polycation copolymers from liposomes. AB - Triggered release of adsorbed polymers from liposomes enables protection against immune recognition during circulation and subsequent intracellular delivery of DNA. Polycationic blocks, poly[2-(dimethylamino) ethyl methacrylate] (DMAEMA) (0.8, 3.1, 4.9, or 9.8 kg/mol) or polylysine (K) (3 kg/mol), act as anchors for poly(ethylene glycol) (PEG) (2 or 5 kg/mol) protective blocks. In addition, a copolymer with 15 strictly alternating blocks of PEG (2 kg/mol) and cationic amine sites was evaluated as a protective coating. Incorporation of 1,2-dioleoyl 3-dimethylammonium-propane, a titratable lipid with a pKa of approximately 6.7, allows the liposome's net charge to increase as the pH shifts from 7.4 in the bloodstream to 5.5 in the endosome. The increased net liposome cationicity results in decreased cationic polymer adsorption. The EMPEG113-DMAEMA31 and EMPEG113-DMAEMA62 association constants decrease from 3.1 and 6.2 (mg/m(2))/(mg/ml) at pH 7.4 to 1.7 and 3.2 (mg/m(2))/(mg/ml) at pH 5.5, respectively. However, EMPEG45-DMAEMA5, EMPEG45-DMAEMA20, and EMPEG45-N-DP15 did not show a strong response to changes in pH. Cationic polymer adsorption exceeds calculated values for liposome neutralization, resulting in adsorption profiles in the brush regime. PMID- 16380162 TI - Generation of a long-lasting, protective, and neutralizing antibody response to the ranavirus FV3 by the frog Xenopus. AB - Xenopus serves as an experimental model to evaluate the contribution of adaptive immunity in host susceptibility to emerging ranaviral diseases that may contribute to amphibian population declines. It has been previously shown that following a secondary infection with the ranavirus frog virus 3 (FV3), adult Xenopus more rapidly clear FV3 and generate specific anti-FV3 IgY antibodies. We have further evaluated the potency and persistence of the Xenopus antibody response against FV3. Frogs inoculated with FV3 (without adjuvant) up to 15 months after priming produce specific, thymus-dependent anti-FV3 IgY antibodies detectable from 10 days to 8 weeks post-infection. These antisera from boosted frogs are neutralizing in vitro and provide partial passive protection to susceptible larvae when they are injected a few minutes before FV3 inoculation. These results with Xenopus suggest that other anuran amphibians are likely to develop effective long-lasting protective humoral immunity after an initial viral exposure. PMID- 16380163 TI - Development of a framework for sustainable uses of resources: more paper and less plastics? AB - Taiwan's EPA has implemented a new guideline called the "Plastic Products Restriction Policy", prohibiting some industries to use plastics as packaging materials for the sake of sustainable use of resources. The significant effect resulting from this policy is the substitution of plastic products with paper products. Is this policy beneficial to achieve future sustainability? I attempt to analyze the resource choice between renewable resources and exhaustible resources for production of final products and services in case of exhaustion of natural resources. In this paper, I develop a framework to examine the dynamic responsiveness of a socio-economical system in facing a continual depletion of natural resources provided by an environmental system. In this framework, the status of an environmental system in terms of carrying capacity is affected by the cumulative impacts caused from human activities, including environmental pollution and resource exploitation. Conversely, it also affects the growth of renewable resources. This framework can serve as a guideline to construct indicators to measure the status of the environmental system and the socio economical system in order to support a policy planner that formulates an appropriate environmental policy. Based on this framework, I also develop a mathematical model to determine the optimal ratio of resources choice between renewable resources and exhaustible resources. PMID- 16380164 TI - From endothelium-derived hyperpolarizing factor (EDHF) to angiogenesis: Epoxyeicosatrienoic acids (EETs) and cell signaling. AB - Epoxyeicosatrienoic acids (EETs) are generated from arachidonic acid by cytochrome P450 (CYP) epoxygenases. The expression of CYP epoxygenases in endothelial cells is determined by a number of physical (fluid shear stress and cyclic stretch) and pharmacological stimuli as well as by hypoxia. The activation of CYP epoxygenases in endothelial cells is an important step in the nitric oxide and prostacyclin (PGI2)-independent vasodilatation of several vascular beds and EETs have been identified as endothelium-derived hyperpolarizing factors (EDHFs). However, in addition to regulating vascular tone, EETs modulate several signaling cascades and affect cell proliferation, cell migration, and angiogenesis. Signaling molecules modulated by EETs include tyrosine kinases and phosphatases, mitogen-activated protein kinases, protein kinase A (PKA), cyclooxygenase (COX) 2, and several transcription factors. This review summarizes the role of CYP derived EETs in cell signaling and focuses particularly on their role as intracellular amplifiers of endothelial cell hyperpolarization as well as in cell proliferation and angiogenesis. The angiogenic properties of CYP epoxygenases and CYP-derived EETs implicate that these enzymes may well be accessible targets for anti-angiogenic as well as angiogenic therapies. PMID- 16380165 TI - Personality and reports of suicide ideation among depressed adults 50 years of age or older. AB - BACKGROUND: Accuracy in patient reports of suicide ideation is a concern in clinical assessment, given that some patients deny suicide ideation even when suicidal. Despite this concern, there is little research on the psychological processes driving reported suicide ideation in at-risk patients. METHODS: A cross sectional design was used to examine the association of personality and suicide ideation in a clinical sample of 134 depressed adults 50 years and older. Patients completed a structured diagnostic interview, an interviewer-rated measure of current suicide ideation and self-report measures of personality (NEO Personality Inventory Revised; NEO-PI-R) and hopelessness. The main outcome variable in logistic regression analyses was suicide ideator status; covariates included comorbid psychopathology, hopelessness and physical illness burden. Predictors were Neuroticism and Openness to Experience (OTE) scores on the NEO-PI R. RESULTS: Elevated OTE and neuroticism were associated with suicide ideation in unadjusted analyses; OTE was also associated with suicide ideation in adjusted regression analyses. LIMITATIONS: This study used a cross-sectional methodology with depressed patients 50 years or older; it is possible that patients' depression severity may have influenced their responses to personality measures. Prospective studies of personality vulnerability to future suicide ideation are warranted. CONCLUSIONS: Elevated neuroticism increases the likelihood of reporting suicide ideation, just as it may enhance risk for suicidal behavior and death by suicide. The pattern for openness is markedly different. Although elevated openness increases the likelihood of reporting suicide ideation, previous research has shown that it may decrease risk of death by suicide, suggesting that the personality-mediated expression of suicide ideation may be adaptive in certain contexts. In contrast, low levels of openness may mute reports of suicide ideation in at-risk patients and confer risk for poor outcomes by potentially undermining clinician vigilance. PMID- 16380166 TI - Immunisation of the male tammar wallaby (Macropus eugenii) with spermatozoa elicits epididymal antigen-specific antibody secretion and compromised fertilisation rate. AB - Immunocontraception has been proposed as an effective and humane means of controlling overabundant kangaroo populations in Australia. We have examined the feasibility of using a sperm-based vaccine for this purpose using a model macropod species, the tammar wallaby (Macropus eugenii). This study has demonstrated immunocontraception in a marsupial species following immunisation of males with homologous spermatozoa. Serum anti-sperm IgG titres were associated with a significant reduction in fertilisation rates following mating with superovulated female wallabies. Antigen-specific IgG penetrated the reproductive tract at the rete testis and bound spermatozoa in vivo. IgG was detected bound to the acrosome and midpiece regions of both epididymal and ejaculated spermatozoa. The absence of adverse testicular pathology and sperm movement effects suggests that contraception may have been achieved by antibody-mediated blocking of sperm surface antigens essential for fertilisation. This study demonstrates that a contraceptive vaccine targeting sperm antigens has potential for fertility control in male macropods. PMID- 16380167 TI - Spermatogenic cells distal to the blood-testis barrier in rats lack C3 convertase regulators and may be at risk of complement-mediated injury. AB - On most tissues, multiple membrane complement regulators (CReg) protect self cells from damage by complement. An exception is the brain, where the blood-brain barrier provides a protected environment within which cells survive with little or no protection from complement. The testis has a functionally similar structure, the blood-testis barrier (BTB). Here, we have investigated the expression of C3/C5 convertase CReg and C3 in the normal rat testis at different ages and different spermatogenetic stages, as well as in rats in which spermatogenesis and the BTB were impaired due to a developmental deficit. Immature testis, prior to BTB formation at puberty, displayed broad expression of the ubiquitous rodent CReg Crry on all elements and no expression of CD46 or CD55. Within days of BTB formation, CReg expression was dramatically altered; Crry was expressed only in the spermatogenetic cells external to the BTB in basal layers of adult seminal epithelium. Spermatogenic cells immediately distal to the BTB at first expressed no C3/C5 convertase regulators but later acquired expression of CD46 and CD55. Staining for C3 was widespread pre-puberty, but absent distal to the BTB in mature rats. In rats with defects in spermatogenesis and BTB integrity, expression patterns of CReg and C3 resembled those in pre pubertal normals. The relative paucity of CReg and absence of C3 synthesis distal to the BTB suggest the presence of a complement-protected environment analogous to that described in the brain, and suggest also that cells enclosed by the BTB may be susceptible to complement damage when the barrier is breached. PMID- 16380168 TI - Cytokine profile of iron-laden macrophages: implications for cellular magnetic resonance imaging. AB - Superparamagnetic iron oxide (SPIO/USPIO) particles are a promising new tool to label cells for in vivo monitoring of their migration into the nervous system by magnetic resonance imaging (MRI). Upon systemic application, SPIO/USPIO particles are preferentially internalized by macrophages. It is unclear whether this affects their immunological profile. We tested the cytokine production of rat and mouse macrophages in vitro and found that internalization of SPIO/USPIO shifted macrophages towards an anti-inflammatory, less responsive phenotype by enhancing interleukin (IL)-10 and inhibiting tumor necrosis factor (TNF)-alpha production. During macrophage interaction with T-cells IL-12p40 secretion was inhibited. Based on our in vitro findings, potential immunomodulatory effects of SPIO/USPIO particles in vivo warrant further investigation. PMID- 16380169 TI - CD24 and myosin light polypeptide 2 are involved in prevention of experimental autoimmune encephalomyelitis by myelin basic protein-pulsed dendritic cells. AB - We previously demonstrated that injection of myelin basic protein-pulsed (MBP pulsed)--but not of unpulsed--autologous bone marrow-derived dendritic cells (DC) efficiently prevents experimental autoimmune encephalomyelitis (EAE) in Lewis rats. To define the molecules involved, we used 3 groups of rats pretreated subcutaneously with MBP-DC, or unpulsed DC, or PBS (control EAE). Four weeks later, all rats were immunized with encephalitogenic MBP peptide and adjuvant. Microarray analyses were done to screen for genes that differ among the 3 groups. Based on microarray analysis data, we used real-time PCR to measure expression of six probably involved genes in draining lymph node cells obtained on day 0, day 7 and day 14 post immunization (p.i.). Two of these 6 genes were consistently altered in both microarray analyses and RT-PCR. They are CD24 antigen being persistently low, and myosin light polypeptide 2 (Myl2) being high in the acute immune response in MBP-DC pretreated rats that develop resistance to EAE. These two genes could be targeted to treat EAE and, possibly, multiple sclerosis. PMID- 16380170 TI - Response to Kitazawa and Wolpert: Rhythmicity, randomness and synchrony in climbing fiber signals. PMID- 16380172 TI - The effects of polycyclic aromatic hydrocarbons on the immune system of fish: a review. AB - Polycyclic aromatic hydrocarbons are an important class of environmental pollutants that are known to be carcinogenic and immunotoxic. This review summarizes the diverse literature on the effects of these pollutants on innate and acquired immunity in fish and the mechanism of PAH-induced immunotoxicity. Among innate immune parameters, many authors have focused on macrophage activities in fish exposed to polycyclic aromatic hydrocarbons. Macrophage respiratory burst appears especially sensitive to polycyclic aromatic hydrocarbons. Among acquired immune parameters, lymphocyte proliferation appears highly sensitive to polycyclic aromatic hydrocarbon exposure. However, the effects of polycyclic aromatic hydrocarbons on both specific and non-specific immunity are contradictory and depend on the mode of exposure, the dose used or the species studied. In contrast to mammals, fewer studies have been done in fish to determine the mechanism of polycyclic aromatic hydrocarbon-induced toxicity. This phenomenon seems to implicate different intracellular mechanisms such as metabolism by cytochrome P4501A, binding to the Ah-receptor, or increased intracellular calcium. Advances in basic knowledge of fish immunity should lead to improvements in monitoring fish health and predicting the impact of polycyclic aromatic hydrocarbons on fish populations, which is a fundamental ecotoxicological goal. PMID- 16380173 TI - Alternative promoters and renal cell-specific regulation of the mouse type IIa sodium-dependent phosphate cotransporter gene. AB - The type IIa sodium-dependent phosphate cotransporter (NPT2a) expressed in renal proximal tubules represents an important determinant in maintaining inorganic phosphate (Pi) homeostasis. In the present study, we identified two variant transcripts of the mouse NPT2a gene, Npt2a-v1 and Npt2a-v2, characterized by the presence of alternative first exons (either exon 1A or exon 1B). The chromosomal structure analysis revealed that the Npt2a gene comprises of two promoters (promoters 1 and 2) and 14 exons, and spans approximately 17 kb. Quantitative PCR analysis showed that renal mRNA levels of both the variants markedly decreased in X-linked vitamin D-resistant hypophosphatemic rickets (Hyp) mice compared to normal littermates. Interestingly, transcriptional activity of a reporter gene, containing Npt2a promoters 1 and 2, was renal cell-specifically increased by 1alpha, 25(OH)2D3 and its analogs. The deletion analysis revealed that the CAAT box in the Npt2a promoter 2 is important for the 1alpha, 25(OH)2D3-dependent renal cell-specific activation of the reporter gene. These data suggested that two alternative promoters control the renal expression of Npt2a gene and both Npt2a variant transcripts are down regulated in Hyp mice. PMID- 16380174 TI - Functional analysis of two regulatory regions of the human Na+ -dependent vitamin C transporter 2, SLC23A2, in human vascular smooth muscle cells. AB - Uptake of vitamin C occurs through the Na+ -dependent vitamin C transporters (SVCT1 and 2), the products of two separate genes. In cultured human vascular smooth muscle cells (hVSMC), we found expression of only the hSVCT2 transcript and identified an additional 5'-UTR transcript variant that we termed exon 1b, in addition to the previously described exon 1a. We cloned and tested the promoter functionality of the two genomic regions of the hSVCT2 upstream of these alternative first exons in hVSMC. Both demonstrated activity, and deletion constructs demonstrated that the minimal promoter regions were within approximately 100 bp relative to their adjacent exons. PMID- 16380175 TI - Inflammation-responsive transcription factors SAF-1 and c-Jun/c-Fos promote canine MMP-1 gene expression. AB - Matrix metalloproteinase-1 (MMP-1) has been implicated in the pathogenesis of osteoarthritis (OA) due to its ability to degrade extracellular matrix component of the joint cartilage tissue that cushions the bone from frictional damage. Canine hip dysplasia, a developmental orthopedic disease which results in arthritic condition as is seen in human OA is an excellent system to study the involvement of MMP-1 in the pathogenesis of OA. To date, however, no report is available regarding canine MMP-1 promoter and the regulatory mechanism by which increased synthesis of MMP-1 protein might be regulated. To gain an insight, we have investigated the promoter region of canine MMP-1. MMP-1 synthesis in the resident cells of arthritic joints is regulated via two major cytokines, IL-1beta and TNF-alpha. By using a series of progressively deleted reporter constructs, multiple cytokine-responsive elements were identified in the proximal promoter region of canine MMP-1. These include DNA-binding elements of AP-1 and SAF-1 transcription factors. Mutation of AP-1 or SAF-1 element resulted in marked reduction in the cytokine responsiveness of MMP-1 promoter. We show that AP-1 and SAF-1 DNA-binding activities are increased in cytokine-stimulated cells as well as in osteoarthritic cartilage tissues. In correlation, immunohistochemical analysis indicated higher levels of MMP-1, SAF-1 and AP-1 proteins in osteoarthritic but not in the normal cartilage tissue. These results show that induction and activation of AP-1 and SAF-1 transcription factors are involved in the regulation of MMP-1 expression in the chondrocytes which could be used as therapeutic targets to combat pathogenesis of OA. PMID- 16380176 TI - E unum pluribus: multiple proteins from a self-processing polyprotein. AB - Many applications of genetic engineering require transformation with multiple (trans)genes, although to achieve these using conventional techniques can be challenging. The 2A oligopeptide is emerging as a highly effective new tool for the facile co-expression of multiple proteins in a single transformation step, whereby a gene encoding multiple proteins, linked by 2A sequences, is transcribed from a single promoter. The polyprotein self-processes co-translationally such that each constituent protein is generated as a discrete translation product. 2A functions in all the eukaryotic systems tested to date and has already been applied, with great success, to a broad range of biotechnological applications: from plant metabolome engineering to the expression of T-cell receptor complexes, monoclonal antibodies or heterodimeric cytokines in animals. PMID- 16380177 TI - Response to Hammer et al.: Tuning genetic control--importance of thorough promoter characterization versus generating promoter diversity. PMID- 16380178 TI - Cancer gene-therapy: clinical trials. AB - The objective of gene therapy for the treatment of cancer is to kill tumour cells but preserve normal tissue; therefore, the ideal gene therapy agent would be targeted for specific transduction of tumour cells and have specificity in its cytotoxic action. A variety of strategies to achieve these aims have demonstrated promising results in the laboratory, including enzyme-pro-drug activating systems, correction of genetic mutations contributing to the malignant phenotype and stimulation of a T-cell-mediated anti-tumour immune response. The key to the success of all these strategies is an effective vector that can direct appropriate expression of the therapeutic gene. Viruses have many properties that can be adapted to achieve this therapeutic endpoint; furthermore, they can be engineered to replicate selectively in cancer cells and lyse them. The challenge now is to translate these features into effective therapies that can supplement or supplant existing treatments. PMID- 16380179 TI - Therapeutic possibilities of plasmonically heated gold nanoparticles. AB - Nanoparticles of gold, which are in the size range 10-100 nm, undergo a plasmon resonance with light. This is a process whereby the electrons of the gold resonate in response to incoming radiation causing them to both absorb and scatter light. This effect can be harnessed to either destroy tissue by local heating or release payload molecules of therapeutic importance. Gold nanoparticles can also be conjugated to biologically active moieties, providing possibilities for targeting to particular tissues. Here, we review the progress made in the exploitation of the plasmon resonance of gold nanoparticles in photo thermal therapeutic medicine. PMID- 16380180 TI - Scientific Citizenship and good governance: implications for biotechnology. AB - In the wake of public distrust regarding biotechnology, it has been suggested that the debate should be moved "upstream", whereby the public help to set research priorities. Although many scientists see this as an illogical reaction to a loss of faith in science, we argue that the boundaries between science and its technological applications have become blurred and this produces conflicts of interests that have led to this crisis of trust. Furthermore, this distrust is also a crisis in governance that calls for a new open and democratic approach to scientific research. We propose that the concept of Scientific Citizenship, based on good governance, will help to restore public trust and bridge the gap between science and the society that it serves. Integral to this is the suggestion that the governance of science forms part of the training for scientists. PMID- 16380181 TI - Interface of biotechnology and ecology for environmental risk assessments of transgenic fish. AB - Genetically engineered fish with enhanced phenotypic traits have yet to be implemented into commercial applications. This is partly because of the difficulties in reliably predicting the ecological risk of transgenic fish should they escape into the wild. The ecological consequences of the phenotypic differences between transgenic and wild-type fish, as determined in the laboratory, can be uncertain because of genotype-by-environment effects (GXE). Additionally, we are limited in our ability to extrapolate simple phenotypes to the complex ecological interactions that occur in nature. Genetic background can also shape the phenotypic effects of transgenes, which, over time and among different wild populations, can make risk assessments a continuously evolving target. These uncertainties suggest that assessments of transgenic fish in contained facilities need to be conducted under as wide a range of conditions as possible, and that efficacious physical and biological containment strategies remain as crucial approaches to ensure the safe application of transgenic fish technology. PMID- 16380183 TI - Yeast ecology in French cider and black olive natural fermentations. AB - In this study, rDNA ITS restriction analysis was used to identify yeasts from two naturally fermented products: French ciders and black olives. In cider musts and bottled ciders, the PCR-RFLP method generated 15 different ITS/RFLP profiles for a total of 208 isolates. The predominant yeasts corresponded to Saccharomyces bayanus, Saccharomyces cerevisiae, Lachancea cidri and Dekkera anomala. Three identified species: Candida sake, Candida tropicalis and Kluyveromyces marxianus had never been described before in ciders. For the black olive fermentation, the method allowed for identification of 11 profiles for a total of 137 isolates. A sequential apparition of yeasts was observed with Pichia anomala, Candida boidinii and Debaryomyces etchellsii being the predominant species. Four isolates did not correspond to any known species based on the sequencing of the D1/D2 region of the 26S rRNA gene. By using the rDNA ITS method, valuable information on yeast population biodiversity and dynamics in the naturally fermented food products studied was obtained. PMID- 16380182 TI - Wound healing morbidity in STS patients treated with preoperative radiotherapy in relation to in vitro skin fibroblast radiosensitivity, proliferative capacity and TGF-beta activity. AB - BACKGROUND AND PURPOSE: In a recent study, we demonstrated that the ability of dermal fibroblasts, obtained from soft tissue sarcoma (STS) patients, to undergo initial division in vitro following radiation exposure correlated with the development of wound healing morbidity in the patients following their treatment with preoperative radiotherapy. Transforming growth factor beta (TGF-beta) is thought to play an important role in fibroblast proliferation and radiosensitivity both of which may impact on wound healing. Thus, in this study we examined the interrelationship between TGF-beta activity, radiosensitivity and proliferation of cultured fibroblasts and the wound healing response of STS patients after preoperative radiotherapy to provide a validation cohort for our previous study and to investigate mechanisms. PATIENTS AND METHODS: Skin fibroblasts were established from skin biopsies of 46 STS patients. The treatment group consisted of 28 patients who received preoperative radiotherapy. Eighteen patients constituted a control group who were either irradiated postoperatively or did not receive radiation treatment. Fibroblast cultures were subjected to the colony forming and cytokinesis-blocked binucleation assays (low dose rate: approximately 0.02 Gy/min) and TGF-beta assays (high dose-rate: approximately 1.06 Gy/min) following gamma-irradiation. Fibroblast radiosensitivity and initial proliferative ability were represented by the surviving fraction at 2.4 Gy (SF(2.4)) and binucleation index (BNI), respectively. Active and total TGF-beta levels in fibroblast cultures were determined using a biological assay. Wound healing complication (WHC), defined as the requirement for further surgery or prolonged deep wound packing, was the clinical endpoint examined. RESULTS: Of the 28 patients treated with preoperative radiotherapy, 8 (29%) had wound healing difficulties. Fibroblasts from patients who developed WHC showed a trend to retain a significantly higher initial proliferative ability after irradiation compared with those from individuals in the treatment group with normal wound healing, consistent with the results of our previous study. No link was observed between fibroblast radiosensitivity and WHC. Neither active nor total TGF-beta levels in cultures were significantly affected by irradiation. Fibroblast proliferation in unirradiated and irradiated cultures, as well as radiosensitivity, was not influenced by TGF-beta content. TGF-beta expression in fibroblast cultures did not reflect wound healing morbidity. CONCLUSIONS: These data are consistent with our previous study and combined the results suggest that in vitro fibroblast proliferation after irradiation may be a useful predictor of wound healing morbidity in STS patients treated with preoperative radiotherapy. TGF-beta levels in culture do not predict WHC, suggesting that the role of TGF beta in wound healing is likely controlled by other in vivo factors. PMID- 16380184 TI - Both arginine and fructose stimulate pH-independent resistance in the wine bacteria Oenococcus oeni. AB - The wine bacteria Oenococcus oeni has to cope with harsh environmental conditions including an acidic pH, a high alcoholic content, and growth inhibitory compounds such as fatty acids, phenolic acids and tannins. So how can O. oeni bacteria naturally present on the surface of grape berries acquire a natural resistance that will alleviate the effect of wine stresses? One mechanism displayed by O. oeni and many other bacteria against the damaging effects of acid environments is arginine consumption through the arginine deiminase pathway. Various studies have shown that the bacterial protection conferred by arginine depends on the rise in pH associated with ammonia production. However, many experimental results disagree with this point of view. The aim of this study was to clarify the protective effect of arginine on O. oeni stress adaptation. Is it only by increasing the pH through ammonia production that this effect is triggered, or does stimulation of appropriate cellular responses play an additional role? This study shows that: (a) arginine in combination with fructose triggers the expression of a subset of genes which are also stress-responsive; (b) cultivation of O. oeni in a fructose- and arginine-supplemented medium prior to wine exposure protects bacteria against subsequent wine shock, and (c) this acquired stress resistance is independent of pH. PMID- 16380185 TI - Distribution of newly described enterotoxin-like genes in Staphylococcus aureus from food. AB - Extensive analysis of the Staphylococcus aureus genome has allowed the identification of new genes encoding enterotoxin-like superantigens (SEls). Some of these are thought to be involved in staphylococcal food poisoning, while others do not elicit any emetic effect. The potential impact of these members of the enterotoxin-like family on the human organism seems to rely mainly on their superantigenic activity. In this paper the distribution of the genes coding for enterotoxin-like superantigens in S. aureus isolated from food was studied. Fifty isolates of S. aureus were examined and 27 were shown to be enterotoxigenic. Only 9 of the 27 strains carried genes encoding enterotoxins SEA-SEE. In 18 SEA-SEE negative strains the presence of newly described enterotoxin genes was detected. All SEA-SEE-positive strains simultaneously carried genes of new SEls. We show that the gene encoding SElH (staphylococcal enterotoxin-like enterotoxin H) was the most frequently detected (n=14), while genes encoding SElI together with SElG accompanied by the other genes of the egc locus were detected in three strains. We also detected the presence of three less investigated genes: sep, sel, and sek. These genes were present in eight, two, and one isolate, respectively. In one strain, sep was accompanied by genes of other SEls, while in the remaining seven it was the only enterotoxin-like gene detected. The high prevalence of newly discovered enterotoxin genes, including the genes encoding emetic toxins, was demonstrated in food-derived strains. This supports the need for additional work on its role in food poisoning and, alternatively, to monitor its presence in S. aureus isolated from food. Our results suggest that yet unknown genetic elements encoding enterotoxin genes may exist. PMID- 16380186 TI - Co-culture experiments demonstrate the usefulness of Lactobacillus sakei 10A to prolong the shelf-life of a model cooked ham. AB - This study investigated the usefulness of two selected lactic acid bacteria, Lactobacillus sakei subsp. carnosus (10A) and the lactocin S producing L. sakei 148 (LS5), to extend the shelf-life of cooked meat products. The interaction between these potential protective cultures and the spoilage organisms, Leuconostoc mesenteroides (LM4) and Brochothrix thermosphacta (BT1), were examined in co-culture studies on a model cooked ham product at 7 degrees C under vacuum packaged conditions. Furthermore, the influence of the glucose content of the model cooked ham on the interaction phenomena was investigated. When artificially contaminating the model cooked ham with BT1 at 10(2) cfu/g in combination with 10A at 10(5) cfu/g, the growth of BT1 was significantly slower compared to a simultaneous mono-culture experiment. In a similar experiment with LM4, LM4 reached a level of 10(7) cfu/g +/-14 days later when LM4 grew together with 10A compared to its growth in mono-culture. The lactocin S producing LS5 did not demonstrate an inhibitory action towards LM4 or BT1 and is therefore not useful as protective culture on cooked meat products. The glucose level of the model cooked ham had no influence on the observed antagonistic interactions of 10A towards LM4 or BT1, indicating that the action of the biopreservative 10A in cooked meat products is independent of the substrate glucose. PMID- 16380187 TI - Ultrasound radiation force enables targeted deposition of model drug carriers loaded on microbubbles. AB - A novel drug delivery vehicle that specifically targets using ultrasound radiation force (USRF) and biotin-avidin interactions is presented. Model vehicles consist of avidinated fluorescent nanobeads bound directly to the biotinylated lipid shells of preformed microbubbles. USRF was used to deflect the vehicle from the center of flow to a tube surface in order to facilitate receptor ligand mediated adhesion. At wall shear stress levels commensurate with venous and arterial flow, USRF was used to direct the vehicles to a biotinylated tube surface. Subsequent high-pressure pulses fragmented the carrier, and molecular interactions induced deposition of the nanobeads on the wall. Targeting of nanobeads to the tube was molecularly specific and dependent on, in order of importance, vehicle concentration, wall shear stress, nanobead size, and insonation time. The observation that portions of the microbubble lipid monolayer shell remain attached to adherent nanobeads is important for future consideration of drug transport mechanisms. This versatile method of delivery is shown to enable targeted deposition of nanoparticles in shear flow and could be modified to carry therapeutic agents for controlled release in targeted delivery applications. PMID- 16380188 TI - Characterization of the transdermal transport of flurbiprofen and indomethacin. AB - Transdermal permeation of two types of NSAIDs, [(3)H] flurbiprofen and [(14)C] indomethacin, was examined by use of the Ussing-type chamber method. We found that the transdermal permeability in the absorptive direction (P(abs)) of [(3)H] flurbiprofen was significantly higher than that of [(14)C] indomethacin. A lower pH (5.0) on the epidermal side increased the accumulation and the P(abs) of [(3)H] flurbiprofen (18-fold and 50-fold, respectively) and [(14)C] indomethacin (18-fold and 22-fold, respectively), compared with pH 7.4. Coadministration of unlabeled flurbiprofen and indomethacin increased P(abs) of [(3)H] flurbiprofen and [(14)C] indomethacin, respectively, in a concentration-dependent manner. Similar high-affinity transport was also observed in the uptake of [(3)H] flurbiprofen by human epidermal keratinocytes (HEK001 cells). RT-PCR analysis revealed the expression of mRNA of numerous transporters including MRP, OATP, MCT and OCTN family members in hairless mouse skin, human skin and HEK001 cells. These findings support the novel hypothesis that transdermal permeation of NSAIDs is mediated by saturable transport mechanisms, which may be candidates as targets for transdermal delivery of drugs. PMID- 16380189 TI - Preparation and characterization of protein-loaded N-trimethyl chitosan nanoparticles as nasal delivery system. AB - In this study, the potential of N-trimethyl chitosan (TMC) nanoparticles as a carrier system for the nasal delivery of proteins was investigated. TMC nanoparticles were prepared by ionic crosslinking of TMC solution (with or without ovalbumin) with tripolyphosphate, at ambient temperature while stirring. The size, zeta-potential and morphology of the nanoparticles were investigated as a function of the preparation conditions. Protein loading, protein integrity and protein release were studied. The toxicity of the TMC nanoparticles was tested by ciliary beat frequency measurements of chicken embryo trachea and in vitro cytotoxicity assays. The in vivo uptake of FITC-albumin-loaded TMC nanoparticles by nasal epithelia tissue in rats was studied by confocal laser scanning microscopy. The nanoparticles had an average size of about 350 nm and a positive zeta-potential. They showed a loading efficiency up to 95% and a loading capacity up to 50% (w/w). The integrity of the entrapped ovalbumin was preserved. Release studies showed that more than 70% of the protein remained associated with the TMC nanoparticles for at least 3 h on incubation in PBS (pH 7.4) at 37 degrees C. Cytotoxicity tests with Calu-3 cells showed no toxic effects of the nanoparticles, whereas a partially reversible cilio-inhibiting effect on the ciliary beat frequency of chicken trachea was observed. In vivo uptake studies indicated the transport of FITC-albumin-associated TMC nanoparticles across the nasal mucosa. In conclusion, TMC nanoparticles are a potential new delivery system for transport of proteins through the nasal mucosa. PMID- 16380190 TI - Preparation of amoxicillin intragastric buoyant sustained-release tablets and the dissolution characteristics. AB - An intragastric buoyant sustained-release tablet (IGB-T) containing 100 mg of amoxicillin (AMX) was prepared to eradicate gastric Helicobacter pylori. A tablet prepared by compressing the mixture of hydroxypropylcellulose-H (HPC-H), citric acid (17.2 mg), sodium hydrogen carbonate (22.8 mg) and AMX was employed as the basic system for preparing IGB-T. The weight and diameter of the tablets were designed to be about 300 mg and 10 mm, respectively. IGB-T containing 5 mg of AMX and HPC-H (255 mg) was buoyant and showed a sustained-release pattern in water. However, when AMX was increased and HPC-H decreased to maintain the tablet weight (300 mg), there was no apparent sustained-release pattern. To prepare IGB-T containing 50 mg of AMX, the surface of the tablet was coated with HPC-H after a tablet was prepared from the mixture of AMX (50 mg), HPC-H (210 mg), citric acid (17.2 mg), and sodium hydrogen carbonate (22.8 mg). This tablet (IGB-T50-Coating) was buoyant and showed a sustained-release pattern in water. However, to complete IGB-T with 100 mg of AMX, it was necessary not only to coat the surface of the tablet but also to use granulated AMX with a particle size of 300-500 microm (IGB T100-Coating-300-500G). IGB-T100-Coating-300-500G was confirmed to be buoyant for 24 h while maintaining a tablet shape and showed a sustained-release pattern in water and buffer solutions of pH 1.2 and 6.8. PMID- 16380192 TI - MIAME, we have a problem. PMID- 16380191 TI - Reliability and reproducibility issues in DNA microarray measurements. AB - DNA microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the current technology has several limitations. Here we discuss problems related to the sensitivity, accuracy, specificity and reproducibility of microarray results. The existing data suggest that for relatively abundant transcripts the existence and direction (but not the magnitude) of expression changes can be reliably detected. However, accurate measurements of absolute expression levels and the reliable detection of low abundance genes are difficult to achieve. The main problems seem to be the sub optimal design or choice of probes and some incorrect probe annotations. Well designed data-analysis approaches can rectify some of these problems. PMID- 16380193 TI - Combinatorial gene regulation in Plasmodium falciparum. AB - The malaria parasite Plasmodium falciparum has a complicated life cycle with large variations in its gene expression pattern, but it contains relatively few specific transcriptional regulators. To elucidate this paradox, we identified regulatory sequences, using an approach that integrates the sequence conservation among species and the correlation in mRNA expression within a species. Our analysis identified several DNA sequence motifs that are associated with mRNA expression, two of which were previously determined experimentally. We found more putative regulatory sequences per gene in P. falciparum than in other eukaryotes, such as yeast. We propose that Plasmodium uses the few regulatory proteins it has in a combinatorial approach for gene regulation, explaining the relative paucity in regulatory proteins. PMID- 16380195 TI - [Grand types of studies 2: analytical studies]. AB - PURPOSE: This paper describes the hierarchy of analytical studies, from the observational studies to the experimental ones. STATE OF THE ART AND MAIN POINTS: Case-control studies, the most frequently performed among analytical studies, may generate or confirm hypotheses. They are of particular use for rare diseases. Cohort studies aim mainly at confirming hypotheses already tested, and at precisely quantifying the magnitude of effect. Randomisation aims at the elimination of confounding factors, and meta-analysis, at decreasing potential selection biases and increasing power. Those goals may not always be achieved. PERSPECTIVES AND PROJETS: The study power, or the beta error, are major factors to be determined when designing a study, before any attempt of realization. PMID- 16380194 TI - Synthesis and in vitro microbiological evaluation of imidazo(4,5 b)pyridinylethoxypiperidones. AB - A series of imidazo(4,5-b)pyridinylethoxypiperidones was designed, synthesized and characterized for evaluation of potential antibacterial activity against Bacillus subtilis, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa and antifungal activity against Candida albicans-6, Candida albicans, Aspergillus niger, Candida albicans-51 and Aspergillus flavus. Structure-activity relationship led to the conclusion that compound 39 exerted strong in vitro antibacterial activity against Bacillus subtilis and Staphylococcus aureus whereas compounds 38 and 39 displayed promising antifungal activity against Aspergillus flavus. The interesting antimicrobial profile of compound 39 led us to select this derivative for further development. PMID- 16380196 TI - [Grand types of clinical studies]. AB - PURPOSE: This paper provides an overview of the hierarchy between the main descriptives clinical studies, with their respective weaknesses and strength. STATE OF THE ART AND MAIN POINTS: Case-reports and case-series are first described, which do not provide control groups but have been very useful in disease recognition and nosology building. Ecological studies, which are hypothesis generating at the group or population level, but which do not study individuals, and prevalence studies with one simple or a double question, which study individuals within groups but may lead to erroneous conclusions so far are risk factors are involved, are then reviewed. PERSPECTIVES AND PROJECTS: The overview of the descriptive studies leads to the notion of confounding factor, which may be better addressed by analytical studies. PMID- 16380197 TI - A randomised-controlled trial in England of a postnatal midwifery intervention on breast-feeding duration. AB - OBJECTIVE: To determine whether postnatal 'hands off' care by midwives on positioning and attachment of the newborn baby improves breast-feeding duration. DESIGN: Mothers were randomised at the first postnatal feed to receive either care by a midwife trained in the experimental protocol or by a control midwife undertaking routine care. SETTING: Eight wards in four English Midlands hospitals. PARTICIPANTS: 370 primiparous mothers with term babies who intended to breast feed, and could sit out of bed to do so. INTERVENTIONS: Experimental protocol of verbal-only advice on positioning and attachment, delivered at the first postnatal ward feed compared with routine care by a qualified midwife. MAIN OUTCOME MEASURES: Duration of breast feeding up to 17 weeks as assessed by diaries and interviews with mothers and protocol adherence from self-completed checklist by the midwife. The mothers' self-reported experience of care and support before, during and after delivery were assessed at 6 weeks, and feeding outcomes and employment status at 17 weeks. FINDINGS: Experimental group mothers more often held the baby across their lap and received 'hands off advice', but fewer babies in the experimental than control groups attached and fed: 59% (106/180) vs. 67% (118/175), p=0.1. No significant differences were found in the numbers of mothers breast feeding at 6 or 17 weeks in the experimental and control groups (stopped exclusive breast feeding: 76% (130/172) vs. 77% (126/163) at 6 weeks; 96% (167/174) vs. 96% (161/168) at 17 weeks; odds ratio 1.02, 95% CI 0.77 to 1.22; p=0.8; stopped any breast feeding: 35% (61/172) vs. 32% (53/167) at 6 weeks; 63% (109/173) vs. 60% (101/167) at 17 weeks; odds ratio 1.10, 0.84 to 1.45; p=0.5). There were no significant differences in the incidence of problems with breast feeding and care experienced by mothers before or during hospitalisation (other than at the first postnatal ward feed), nor after discharge home. CONCLUSIONS: No significant beneficial effect was found on breast feeding duration of the verbal- only advice on positioning and attachment, perhaps because aspects of the intervention are already within routine UK practice. Other care practices at subsequent feeds may negate benefits of care at earlier feeds. 'Hands off' care at the first feed may be less important to subsequent feeding than achieving a first feed under supervision in the postnatal ward. IMPLICATIONS FOR PRACTICE: Midwives can be trained in a 4-hr workshop to achieve improved knowledge of 'hands off' positioning and attachment care, and these can be translated into clinical practice. Future studies should differentiate the elements of the care that are effective in achieving postnatal feeds, and apply this advice consistently at successive feeds. PMID- 16380198 TI - [Antebrachial flap based on distal perforators of the radial artery: anatomic study of 10 cases]. AB - PURPOSE: The radial forearm flap based on distal perforators of the radial artery is a recent flap, derived from the so-called << chinese-flap >>. The interest of this flap resides in the preservation of the main arteries of the forearm. The aim of this study was to count the distal perforators of the radial artery and to study sub-cutaneous vascular system. MATERIALS AND METHOD: We performed 10 dissections on fresh cadavers, after selective injection of the radial artery with a solution of plasticized resin. The dissection was performed from the palmar flexor fold of the wrist up to 15 cm proximally, allowing to count all of the distal perforators. RESULTS: We determined three different intervals, among which the one located between 2 and 6 cm from the palmar flexor fold of the wrist was the most vascularized. This interval should be considered as the rotation point of the flap. CONCLUSION: This study allowed to make clear and more reliable the operative procedure to raise the flap. This procedure is easy to handle and intended to cover proximal defects of the dorsal aspect of the hand, while sparing the radial artery. PMID- 16380199 TI - Identification of genes and proteins involved in the pleiotropic response to arsenic stress in Caenibacter arsenoxydans, a metalloresistant beta proteobacterium with an unsequenced genome. AB - The effect of high concentrations of arsenic has been investigated in Caenibacter arsenoxydans, a beta-proteobacterium isolated from an arsenic contaminated environment and able to oxidize arsenite to arsenate. As the genome of this bacterium has not yet been sequenced, the use of a specific proteomic approach based on nano-high performance liquid chromatography tandem mass spectrometry (nanoLC-MS/MS) studies and de novo sequencing to perform cross-species protein identifications was necessary. In addition, a random mutational analysis was performed. Twenty-two proteins and 16 genes were shown to be differentially accumulated and expressed, respectively, in cells grown in the presence of arsenite. Two genes involved in arsenite oxidation and one in arsenite efflux as well as two proteins responsible for arsenate reduction were identified. Moreover, numerous genes and proteins belonging to various functional classes including information and regulation pathways, intermediary metabolism, cell envelope and cellular processes were also up- or down-regulated, which demonstrates that bacterial response to arsenic is pleiotropic. PMID- 16380200 TI - Effect of C-terminal truncation on the molecular chaperone function and dimerization of Escherichia coli trigger factor. AB - To examine the role of the C-terminal domain in the chaperone function of trigger factor (TF), a number of truncation mutants were constructed, namely: TF419, TF389, TF380, TF360, TF344, and TF251, in which the C-terminal 13, 43, 52, 72, 88 residues or the entire C-domain were deleted, respectively. Co-expression of mutant chicken adenylate kinase (AK) with TF and the C-terminal truncation mutants was achieved using a plasmid pBVAT that allows expression of TF and AK from a single plasmid. The results show that truncation of the C-terminus of TF has only minor effect on its ability to assist AK refolding in vivo. Further, ribosome-binding experiments indicate that C-terminal truncation mutants can still bind to the ribosome and the presence of the C-terminus may in fact lower the affinity of TF for the ribosome in vivo. This indicates that the C-domain of trigger factor may not be essential for the ribosome-associated molecular chaperone function of TF. However, the purified TF C-terminal truncation mutants had a dramatically reduced ability to assist rabbit muscle GAPDH refolding in vitro and a reduced tendency to dimerize. This shows that the structural integrity of the C-terminus contributes to both the chaperone function of TF and the stability of the dimeric form. PMID- 16380201 TI - Biphasic effect of IL-1beta on the activity of argininosuccinate synthetase in Caco-2 cells. Involvement of nitric oxide production. AB - The expression of the argininosuccinate synthetase gene (ASS), the limiting enzyme of arginine synthesis, was previously shown to be rapidly induced by a short-term (4 h) exposure to IL-1beta in Caco-2 cells [Biochimie, 2005, 403-409]. The present report shows that, by contrast, a long-term (24 h) exposure to IL 1beta inhibited the ASS activity despite an increase in both specific mRNA level and protein amount, demonstrating a post-translational effect. Concerning the mechanism involved, we demonstrate that the inhibiting effect is linked to the production of nitric oxide (NO) induced by IL-1beta. Indeed, the inhibiting effect of IL-1beta was totally blocked in the presence of l-NMMA, an inhibitor of the inducible nitric oxide synthase, or by culturing the cells in an arginine deprived medium. Moreover, a decrease in the ASS activity was induced by culturing the cells in the presence of SNAP, a NO donor. Conversely, blocking the action of NO by antioxidant agents, the stimulatory effect of IL-1beta on ASS activity was restored, as measured at 24 h. Finally, such an inhibiting effect of NO on ASS activity may be related, at least in part, to S-nitrosylation of the protein. The physiological relevance of the antagonistic effects of IL-1beta and NO on ASS is discussed. PMID- 16380203 TI - Ground-based methods reproduce space-flight experiments and show that weak vibrations trigger microtubule self-organisation. AB - The effect of weightlessness on physical and biological systems is frequently studied by experiments in space. However, on the ground, gravity effects may also be strongly attenuated using methods such as magnetic levitation and clinorotation. Under suitable conditions, in vitro preparations of microtubules, a major element of the cytoskeleton, self-organise by a process of reaction diffusion: self-organisation is triggered by gravity and samples prepared in space do not self-organise. Here, we report experiments carried out with ground based methods of clinorotation and magnetic levitation. The behaviour observed closely resembles that of the space-flight experiment and suggests that many space experiments could be carried out equally well on the ground. Using clinorotation, we find that weak vibrations also trigger microtubule self organisation and have an effect similar to gravity. Thus, in some in vitro biological systems, vibrations are a countermeasure to weightlessness. PMID- 16380202 TI - Does glycosylation of melanoma cells influence their interactions with fibronectin? AB - Cell surface integrins, especially those binding to fibronectin (FN), participate in processes of tumor cell invasion and metastasis. Changes in glycosylation of cell surface adhesion proteins are often associated with malignant transformation of cells. In this study we examined the influence of swainsonine (SW) on adhesion, wound healing and haptotactic migration on FN, comparing the responses of different human melanoma cell lines: primary WM35 and metastatic WM9, WM239 and A375. We also examined the role of alpha subunits in adhesion to FN. All of the antibodies inhibited adhesion to FN but with different efficiencies depending on the cell line. Adhesion was mediated mainly by integrin alpha(5)beta(1) (WM9, A375), alpha(3)beta(1) (WM35, A375, WM239). Scratch wound repair was significantly faster on FN-coated wells than on plastic for all cells except for WM9. A375 and WM9 had the greatest migration ability, both expressing the highest level of alpha(5)beta(1) integrin. It seems very likely that adhesion to FN can be accomplished by many different integrins, but for effective migration alpha(5)beta(1) integrin is responsible. Only A375 and WM239 cell lines reacted to SW treatment. In the presence of SW WM239 and A375 cells had 70% and 40% increased adhesion to FN, and their migration was decreased 40% and 50%, respectively. Interestingly, although most of the cell lines share a common profile of integrins, each line interacted with FN differently. They differed mainly in the repertoire of integrins used for adhesion, and in the manner in which glycosylation affected these processes. The influence of SW was observed in two metastatic cell lines indicating the contribution of glycosylation status to the progression of melanoma. The lack of reaction to SW in WM9 cells may suggest that there is a threshold in the expression level of the highly branched N glycans that may influence the adhesion and migration properties of the cell. PMID- 16380204 TI - Dielectric behavior of DNA in water-organic co-solvent mixtures. AB - The radiowave dielectric dispersions of DNA in different water-organic co-solvent mixtures have been measured in the frequency range from 100 kHz to 100 MHz, where the polarization mechanism is generally attributed to the confinement of counterions within some specific lengths, either along tangential or perpendicular to the polyion chain. The dielectric dispersions have been analyzed on the basis of two partially different dielectric models, a continuum counterion fluctuation model proposed by Mandel and a discrete charged site model, proposed by Minakata. The influence of the quality of the solvent on the dielectric parameters has been investigated in water-methanol and water-glycerol mixtures at different composition, by varying the permittivity (m) and the viscosity eta of the solvent phase. The analysis of the dielectric spectra in solvents where electrostatic and hydrodynamic interactions vary with the solvent composition suggests that both the two models are able, in principle, to account for the observed high-frequency dielectric behavior. However, while some certain assumptions are necessary about the polyion structure within the Mandel model, no structural prerequisite is needed within the Minakata model, where the polarization mechanism invoked considers a radial counterion exchange with the outer medium, which is largely independent of the local polyion conformation. PMID- 16380205 TI - Interfacial tension of phosphatidylcholine-phosphatidylserine system in bilayer lipid membrane. AB - The effect of pH of electrolyte solution on the interfacial tension of lipid membrane formed of phosphatidylcholine (PC, lecithin)-phosphatidylserine (PS) system was studied. In this article, three models describing the H+ and OH- ions adsorption in the bilayer lipid surface are presented. In Model I and Model II, the surface is continuous with uniformly distributed functional groups constituting the centres of H+ and OH- ions adsorption while in the other the surface is built of lipid molecules, free or with attached H+ and OH- ions. In these models contribution of the individual lipid molecule forms to interfacial tension of the bilayer were assumed to be additive. In Model III the adsorption of the H+ and OH- ions at the PC-PS bilayer surface was described in terms of the Gibbs isotherm. Theoretical equations are derived to describe this dependence in the whole pH range. PMID- 16380206 TI - Valuing river characteristics using combined site choice and participation travel cost models. AB - This paper presents new welfare measures for marginal changes in river quality in selected English rivers. The river quality indicators used include chemical, biological and habitat-level attributes. Economic values for recreational use of three types of river-upland, lowland and chalk-are presented. A survey of anglers was carried out and using these data, two travel cost models were estimated, one to predict the numbers of trips and the other to predict angling site choice. These models were then linked to estimate the welfare associated with marginal changes in river quality using the participation levels as estimated in the trip prediction model. The model results showed that higher flow rates, biological quality and nutrient pollution levels affect site choice and influence the likelihood of a fishing trip. Consumer surplus values per trip for a 10% change in river attributes range from pound 0.04 to pound 3.93 ( pound 2001) depending on the attribute. PMID- 16380207 TI - Fine epitope mapping of monoclonal antibodies specific to human alpha-synuclein. AB - The neuronal phosphoprotein alpha-synuclein has been increasingly implicated in the pathogenesis of Parkinson's disease (PD) and other neurodegenerative diseases; however, the exact function of alpha-synuclein still remains illusive. Suitable antibodies (Abs) specific for the gene of interest are indispensable for studying biological and immunological properties of the target gene. Here, we report not only the generation and characterization of monoclonal Abs, Syn-1 and Syn-17, against human alpha-synuclein, but also the epitope mapping by using recombinant synuclein family proteins and various GST fusion proteins of human alpha-synuclein domains. Syn-17 recognizes human and rodent alpha-synuclein, and its epitope is localized within residues 97-99 and 101 of alpha-synuclein. In contrast, the Syn-1 epitope is localized in residues 121 and 122 of human alpha synuclein, and Syn-1 recognizes only human but not rodent alpha-synuclein, indicating that it can be utilized as a useful reagent for studying human alpha synuclein transgenic mouse and zebrafish lines. PMID- 16380208 TI - Ca2+-dependent PKC activation mediates menthol-induced desensitization of transient receptor potential M8. AB - In 1950, Hensel and Zotterman reported cooling-induced desensitization of cold receptors by extracellular discharge recordings of cold fibers. Since then, however, its intracellular mechanism has remained unresolved. We studied menthol induced desensitization of cold/menthol receptors (TRPM8, transient receptor potential M8) expressed in HEK cells. TRPM8 desensitization depended on extracellular Ca2+ ions, indicating that Ca2+ influx-induced [Ca2+]i elevation caused the desensitization. We studied whether Ca2+-dependent kinase, PKC, mediated TRPM8 desensitization. PMA, a PKC activator, desensitized TRPM8. Inhibitor of Ca2+-dependent PKC isozymes specifically abolished PMA-induced TRPM8 desensitization. PMA similarly desensitized wild type TRPM8 and mutant TRPM8, in which serine or threonine residues in some putative PKC phosphorylation sites were replaced by alanine. PMA treatment did not induce internalization of TRPM8. As the basis of cooling-induced desensitization of cold receptors, we conclude that cooling-activated TRPM8 causes Ca2+-dependent PKC isozymes to desensitize TRPM8 itself. PMID- 16380210 TI - The discharge of subthalamic neurons is modulated by inhibiting the nitric oxide synthase in the rat. AB - The effects induced on the discharge of subthalamic spontaneously active neurons by inhibiting the enzyme nitric oxide synthase was studied in two groups of urethane-anesthetized rats. In the first group of animals (n = 10), the activity of subthalamic single units was recorded before and after the systemic administration of 7-nitro-indazole (7-NI, 50 mg/kg i.p.), a selective inhibitor of neuronal nitric oxide synthase. In the second group of rats (n = 15), Nomega nitro-L-arginine methyl ester (L-NAME), another inhibitor of nitric oxide synthase, was iontophoretically administered while performing single unit extracellular recordings. The activity of most tested spontaneously discharging neurons (8/10) was influenced by 7-NI administration, which always caused a statistically significant decrease in the firing rate of the responsive cells. In contrast, the iontophoretic administration of L-NAME, although influencing many cells (24/32), did not have univocal effects: in fact, 18 cells were inhibited while 6 neurons were excited in a statistically significant manner. We hypothesize that nitric oxide neurotransmission could exert a tonic modulatory influence upon spontaneously discharging subthalamic neurons, with a prevalent excitatory effect. PMID- 16380211 TI - Applied glycoproteomics--approaches to study genetic-environmental collisions causing protein-losing enteropathy. AB - Protein-losing enteropathy (PLE), the loss of plasma proteins through the intestine, is a life-threatening symptom associated with seemingly unrelated conditions including Crohn's disease, congenital disorder of glycosylation, or Fontan surgery to correct univentricular hearts. Emerging commonalities between these and other disorders led us to hypothesize that PLE develops when genetic insufficiencies collide with simultaneous or sequential environmental insults. Most intriguing is the loss of heparan sulfate (HS) proteoglycans (HSPG) specifically from the basolateral surface of intestinal epithelial cells only during PLE episodes suggesting a direct link to protein leakage. Reasons for HSPG loss are unknown, but genetic insufficiencies affecting HSPG biosynthesis, trafficking, or degradation may be involved. Here, we describe cell-based assays we devised to identify key players contributing to protein leakage. Results from these assays confirm that HS loss directly causes protein leakage, but more importantly, it amplifies the effects of other factors, e.g., cytokines and increased pressure. Thus, HS loss appears to play a central role for PLE. To transfer our in vitro results back to the in vivo situation, we established methods to assess enteric protein leakage in mice and present several genetically deficient strains mimicking intestinal HS loss observed in PLE patients. Preliminary results indicate that mice with haploinsufficient genes involved in HS biosynthesis or HSPG trafficking develop intestinal protein leakage upon additional environmental stress. Our goal is to model PLE in vitro and in vivo to unravel the pathomechanisms underlying PLE, identify patients at risk, and provide them with a safe and effective therapy. PMID- 16380209 TI - Trifluoperazine, an orally available clinically used drug, disrupts opioid antinociceptive tolerance. AB - Calcium/calmodulin dependent protein kinase II (CaMKII) has been shown to play an important role in the generation and maintenance of opioid tolerance. In this study, trifluoperazine was studied for its effect on morphine tolerance in mice. Acute treatment with trifluoperazine (0.5 mg/kg, i.p.) completely reversed the established antinociceptive tolerance to morphine. Pretreatment with trifluoperazine also significantly attenuated the development of antinociceptive tolerance (p<0.01). Morphine induced a significant up-regulation of supraspinal and spinal CaMKII activity in tolerant mice, which was abolished after the pretreatment or acute treatment with trifluoperazine. These data suggested that trifluoperazine was capable of suppressing opioid tolerance, possibly by the mechanism of inhibiting CaMKII. Since trifluoperazine has been safely used as an antipsychotic drug, we propose that the drug should be studied in humans for the prevention and treatment of opioid tolerance and addiction. PMID- 16380212 TI - A clinical forensic scientist in the burns unit: necessity or not? A prospective clinical study. AB - Child abuse by burning is a common means of assault that may be difficult to prove. Even though well trained in burns management physicians on the burn team may not be sufficiently qualified to clarify the medicolegal aspects of the incident. In Turkey, physicians have a responsibility to notify the law offices of injury to children caused by any non-accidental mechanism including neglect. The consequences of false positive and/or false negative reports to the legal offices may be damaging for the care-takers and/or the children. In our study, 239 consecutively hospitalized children with burns were studied prospectively. A clinical forensic scientist and a physician of the burns team interpreted incidents separately so that neither had an idea about the other's diagnoses until the end of the study. There were found to be some differences in the interpretation of the incidents by the clinical forensic scientist and the burns team physician. These differences were described as discordant diagnoses. The physician accepted 99 (41.4%) of the incidents as an accident while only 66 (27.6%) of the victims were labelled as accidents by the forensic scientist. A multivariate analysis identified low socioeconomic status, aged between 3 and 6 years, and a family size of more than six as independent variables significantly associated with discordant diagnoses (p = 0.0388, 0.0001, 0.0203, respectively). As a conclusion, we suggest that to minimize the likelihood of a legally inaccurate diagnosis, a clinical forensic scientist seems to be necessary in the emergency department. PMID- 16380213 TI - Enhanced healing of full-thickness burn wounds using di-rhamnolipid. AB - The aim of this study was to investigate the properties of di-rhamnolipid [alpha L-rhamnopyranosyl-(1-2)-alpha-L-rhamnopyranosyl-3-hydroxydecanoyl-3 hydroxydecanoic acid, also referred to as di-rhamnolipid BAC-3] relating to the process of cutaneous wound healing. Di-rhamnolipid was prepared in a eucerin ointment and applied topically on full-thickness burn wounds in normal Sprague Dawley rats covering 5% of the total body surface area. The rate of wound closure was measured over the period of 45 days. The collagen content was evaluated microscopically, by performing densitometric analysis on Verhoeff's stained histopathological slides of wound biopsies taken at the end of 45th day of di rhamnolipid treatment. Di-rhamnolipid toxicity was assessed with the subcutaneous multi-dose study in Swiss-Webster mice. The treatment of full-thickness-burn wounds with topical 0.1% di-rhamnolipid accelerated the closure of wounds on day 21 of the treatment by 32% compared to the control (p < 0.05). On day 35, the wounds closed in all animals-treated with 0.1% di-rhamnolipid ointment while some rats in the control group had open wounds on days 35 and even 45. Histologic comparisons have shown that di-rhamnolipid significantly decreased collagen content in burn wounds (47.5%, p < 0.05) as compared to the vehicle-treated (control) wounds. Di-rhamnolipid was well-tolerated. The results of this study raise the possibility of potential efficacy of di-rhamnolipid in accelerating normal wound healing and perhaps in overcoming defects associated with healing failure in chronic wounds. PMID- 16380214 TI - Application of cerium nitrate-silver sulphadiazine allows for postponement of excision and grafting. AB - Early excision and grafting cannot always be performed due to patient's medical status, and the lack of adequate donor sites. The use of cerium nitrate-silver sulphadiazine cream, which causes the formation of a leather-like eschar with excellent resistance to infection, is an alternative method. In order to postpone operations by using cerium nitrate-silver sulphadiazine, we compared the differences in contamination of the grafted areas between early and delayed excision. Eighteen patients underwent excision and grafting within 5 days post burn and nineteen patients were surgically treated after 5 days. Twelve months later the grafted areas were evaluated. Contamination of the grafted area occurred in 17 patients. No differences in contamination occurred between the early and delayed excision group, 8 versus 9. Also no differences in type of organism cultured and follow-up results were found between the early and delayed excision group. Cerium nitrate-silver sulphadiazine allows surgical treatment to be delayed without an increase of contamination of the grafted area and does not adversely affect the long-term outcome. PMID- 16380215 TI - Leakage of liquefied petroleum gas during motor vehicle refuelling--a new cause of cold injury. PMID- 16380216 TI - Evaluation of a bi-layer wound dressing for burn care I. Cooling and wound healing properties. AB - Severe burns remain a significant cause of morbidity and mortality despite the availability of numerous therapies. We assessed the wound healing and skin cooling properties of a DRDC hydrogel/polyurethane wound dressing using different pre-clinical models. Our results show that 85% of partial-thickness, non contaminated porcine wounds treated with our dressing healed within 6 days. In contrast, 85% of the wounds treated with commercial dressings healed within 8 days. Application of a moist DRDC dressing (to simulate a condition of exudate absorption) on a scald burn covering 25% of the dorsal area in rats reduced skin temperature by 1.70 +/- 0.14 degrees C for 5 min, the skin temperature being comparable to that of control burned rats after 20 min. The application of a moist DRDC dressing did not induce significant differences in body temperatures compared with that of burned animals without dressing coverage throughout the 90 min experiment. While no change in body temperatures were observed when standard dressings (i.e., not pre-moistened) were applied, skin temperature increased gradually. These data show that our dressing is effective in promoting faster healing of the treated wound; and providing a transient, but beneficial cooling effect to the skin contact-site, without the adverse effect of inducing whole body hypothermia. PMID- 16380217 TI - Health and well being of substance use disorder patients with and without posttraumatic stress disorder. AB - While studies link posttraumatic stress disorder (PTSD) to substance use disorders (SUDs), little is known about the health and functional status of patients with such comorbidity. This study examined the health and well being of SUD patients with and without PTSD. Participants were assessed using structured clinical interviews and self-administered questionnaires assessing chronic physical symptoms, and functional status and well being. PTSD was significantly associated with more cardiovascular, neurological, and total chronic physical symptoms. In addition, PTSD was significantly related to poorer functional status and well being, particularly in the mental health realm of functioning. The association between PTSD and mental health component of functional health and well-being remained significant after controlling for chronic physical symptoms and other comorbid psychopathology. PMID- 16380218 TI - Functional characterization of human osteoclast inhibitory peptide-1 (OIP-1/hSca) gene promoter. AB - We have recently identified and characterized the human osteoclast (OCL) inhibitory peptide-1 (OIP-1/hSca), a member of Ly-6 gene family. OIP-1 is an important physiologic regulator of OCL development and bone resorption activity. To determine the molecular mechanisms that regulate OIP-1 gene expression in OCL precursor cells, we isolated and characterized the OIP-1/hSca gene (2 Kb) promoter sequence. IFN-gamma (50 ng/ml) treatment of RAW 264.7 macrophage cells transfected with OIP-1 gene (-1 to -1988 bp relative to transcription start site) promoter-luciferase reporter plasmid demonstrated a significant (4 fold) increase in OIP-1 gene promoter activity. Sequence analysis of OIP-1 gene promoter region further identified a potential Stat-1 binding motif at -1629 to -1639 bp position. Stat-1 specific inhibitor, fludarabine (50 muM) abolished IFN-gamma stimulated OIP-1 gene promoter activity. Electrophoretic mobility shift assay (EMSA) further confirmed activated Stat-1 binding to the OIP-1 gene promoter sequence suggesting that IFN-gamma regulates OIP-1 gene expression in OCL precursor cells through a Stat-1 dependent signaling pathway. We further show that knock-down of TRADD enhances IFN-gamma signaling to increase OIP-1 gene expression in OCL precursor cells. These results should provide insights into the molecular control of OIP-1 gene expression and inhibition of OCL activity in the bone microenvironment. PMID- 16380220 TI - Alu-associated enhancement of single nucleotide polymorphisms in the human genome. AB - Identifying features shaping the architecture of sequence variations is important for understanding genome evolution and mapping disease loci. In this study, high resolution scanning of Alu-centered alignments of the human genome sequences has revealed a striking elevation of the frequency of single nucleotide polymorphisms (SNP) in the body and tail of Alu sequences compared to flanking regions. This enhancement in SNP density is evident for all twenty-four chromosomes, and in both the Alu-body and Alu-tail, which together may be referred to as the Alu SNPs. Reduced levels of Alu-SNPs in the sex chromosomes, especially in the non recombining NRY region of the Y chromosome, are consistent with recombination events playing an important role in the enhancement. The Alu elements are unstable recombination-mutation hotspots in the human genome, and it is suggested that the Alu-SNPs represent a key manifestation of this instability. Variations in Alu-SNPs among the HapMap populations of northern and western European ancestry (CEU), Han Chinese from Beijing (CHB), Japanese from Tokyo (JPT), and Yoruba from Ibadan, Nigeria (YRI) indicate that the Alu-SNPs provide useful sequence markers, in addition to the Alu-insertion polymorphisms themselves, for the delineation of human genome evolution. That Alu-SNP levels are highest in the youngest Alu-Y, intermediate in the Alu-S of intermediate age, and lowest in the oldest Alu-J is consistent with the occurrence of not only genetic drift but also natural selection on the Alu-SNPs. Such evolutionary selection in turn suggests that Alu-SNPs might include potential sites of disease association, and therefore deserve detailed investigation. PMID- 16380219 TI - Identification of novel PPARgamma target genes in primary human adipocytes. AB - Adipogenesis is the process by which undifferentiated precursor cells differentiate into fat laden adipocytes. The nuclear proteins peroxisome proliferator-activated receptors (PPARs) play a central role in adipocyte differentiation. The goals of this study were to identify novel PPARgamma responsive genes and to determine their role in regulating human adipocyte differentiation. Affymetrix profiling of gene expression in human adipocytes identified about 1000 genes that were significantly up-regulated subsequent to induction of differentiation. PPARgamma expression was reduced prior to induction of differentiation using a novel, chemically modified antisense oligonucleotide. Affymetrix microarray profiling of these cells identified 278 statistically significantly down-regulated genes. Eight genes were found to contain previously documented PPARgamma recognition element (PPRE) in their upstream nucleotide (promoter) sequence. Four of these genes are novel and have not previously been characterized. Chromatin immuno-precipitation experiments confirmed the binding of PPARgamma to the PPRE of three of these genes. The ortholog of one of these genes, hypothetical protein FLJ 20920, has previously been reported to be involved in the control of body fat composition in Caenorhabditis elegans. Inhibition of expression of this protein was found to also inhibit differentiation of human adipocytes. MAST/MEME algorithm analysis was used to identify novel commonly occurring sequence motifs in the 5' upstream region of transcripts for subset of down-regulated genes, which were grouped according to their sequence similarities. A number of clusters were identified and the largest cluster contained similar motifs from 26 genes with the literature supporting 7 of the 26 genes as being involved in fatty acid metabolism or PPARgamma interaction. PMID- 16380221 TI - Synonymous codon usage in environmental chlamydia UWE25 reflects an evolutional divergence from pathogenic chlamydiae. AB - Publication of the complete genome sequence for the Acanthamoeba sp. endosymbiont UWE25 has illuminated the evolution history of chlamydiae. In this study, the codon usage bias in UWE25 and five other species of pathogenic chlamydiae was calculated. It was found that genomic composition constraints are the major source of codon usage variation in UWE25. This result is different from the former observation in pathogenic chlamydiae, whose genomic base composition is more unbiased. Four other factors, such as strand-specific mutational bias, natural selection acting at the level of translation, hydropathy level of each protein and the conservation level of amino acids also have influence in shaping the codon usage in these six species to some extent. Further analysis suggests that the high stability of the UWE25 genome partially account for the difference in codon usage pattern between environmental and pathogenic chlamydiae. Moreover, our results imply that the replicational selection pressure in pathogenic chlamydiae is stronger than that in UWE25. Analyzing the codon usage pattern in the environmental chlamydia and comparing it with that of the pathogenic chlamydiae may provide clues how the chlamydiae have evolved from their common ancestor. PMID- 16380222 TI - Evolutionary analysis of a large mtDNA translocation (numt) into the nuclear genome of the Panthera genus species. AB - Translocation of cymtDNA into the nuclear genome, also referred to as numt, has been reported in many species, including several closely related to the domestic cat (Felis catus). We describe the recent transposition of 12,536 bp of the 17 kb mitochondrial genome into the nucleus of the common ancestor of the five Panthera genus species: tiger, P. tigris; snow leopard, P. uncia; jaguar, P. onca; leopard, P. pardus; and lion, P. leo. This nuclear integration, representing 74% of the mitochondrial genome, is one of the largest to be reported in eukaryotes. The Panthera genus numt differs from the numt previously described in the Felis genus in: (1) chromosomal location (F2-telomeric region vs. D2-centromeric region), (2) gene make up (from the ND5 to the ATP8 vs. from the CR to the COII), (3) size (12.5 vs. 7.9 kb), and (4) structure (single monomer vs. tandemly repeated in Felis). These distinctions indicate that the origin of this large numt fragment in the nuclear genome of the Panthera species is an independent insertion from that of the domestic cat lineage, which has been further supported by phylogenetic analyses. The tiger cymtDNA shared around 90% sequence identity with the homologous numt sequence, suggesting an origin for the Panthera numt at around 3.5 million years ago, prior to the radiation of the five extant Panthera species. PMID- 16380223 TI - Identification of paralogous genes of firefly luciferase in the Japanese firefly, Luciola cruciata. AB - Two homologous genes of firefly luciferase, LcLL1 and LcLL2, were cloned from the Japanese firefly Luciola cruciata, and were expressed and characterized. The gene product of LcLL1 had long-chain fatty acyl-CoA synthetic activity, but not luciferase activity. The other gene product of LcLL2 did not show enzymatic activities of acyl-CoA synthetase and luciferase. RT-PCR analysis showed that the transcript of LcLL1 was abundant in larva but very low in adult, while LcLL2 was expressed in both larva and adult. Phylogenetic analysis indicated that LcLL1 and LcLL2 are paralogous genes of firefly luciferase. Recently, we found that CG6178 in Drosophila melanogaster is an orthologue of firefly luciferase and shows fatty acyl-CoA synthetic activity, but not luciferase activity. These results suggest that firefly luciferase might be evolved from a fatty acyl-CoA synthetase by gene duplication in insects. PMID- 16380224 TI - Prenatal diagnosis and management of hydronephrosis. AB - Congenital hydronephrosis is frequently amenable to prenatal diagnosis, often as early as the second trimester. Most clinicians use a renal pelvis anterior posterior (AP) diameter of 4 mm or more prior to 20 weeks of gestation as a threshold for identifying pyelectasis. If mild dilation of the renal pelvis is an isolated finding in the second trimester, evaluation performed later in gestation is used to guide postnatal management. Since the normal renal pelvis dimensions may increase with advancing gestation, thresholds for the diagnosis are larger in the third trimester. Neonatal follow-up is typically recommended only if the fetal renal pelvis diameter exceeds a specified cut-off (e.g. 7 or 10 mm) at or beyond 34 weeks. If the measurement is less, most deem the pyelectasis physiologic or normal. However, it has been suggested that fetuses with early renal pelvis dilation that resolved during pregnancy might also benefit from postnatal surveillance. The newborn evaluation for hydronephrosis may be time consuming, invasive, and costly; however, it can often prevent sequelae from congenital uropathy. PMID- 16380225 TI - The nisin-controlled gene expression system: construction, application and improvements. AB - Lactic acid bacteria are widely used in industrial fermentation. The potential use of these bacteria as homologous and heterologous protein expression hosts has been investigated extensively. The NIsin-Controlled gene Expression system (the NICE system) is an efficient and promising gene expression system based on the autoregulation mechanism of nisin biosynthesis in the Lactococcus lactis. In the NICE system, the membrane-located histidine kinase NisK senses the inducing signal nisin and autophosphorylates, then transfers phosphorous group to intracellular response regulator protein NisR which activates nisA promoter to express the downstream gene(s). The NICE system allows regulated overproduction of a variety of interest proteins by several Gram-positive bacteria, especially L. lactis. The essential elements for system construction, its application for expression of some biotechnologically important proteins and further improvements of this system are discussed. PMID- 16380226 TI - Aromatase inhibitors for early breast cancer therapy: a choice of effective treatment strategies. PMID- 16380228 TI - Evaluation of leptin levels in subjects at risk for type 1 diabetes. AB - Type 1 diabetes (T1DM) is increasing in incidence worldwide, particularly in young children. Studies have suggested that weight gain in early childhood may play a role in determining disease risk, with increased risk in children who have gained more weight. We hypothesized that leptin may be involved by promoting a Th1-type immune response in individuals at risk for the development of T1DM. Insulin, GAD65, and IA-2 autoantibodies and leptin levels were measured in relatives of T1DM patients and in control subjects. After adjusting for HLA risk and age, autoantibody-positive relatives were compared with sex- and BMI-matched autoantibody-negative control subjects. BMI-matched individuals had similar leptin levels, and the leptin levels increased with increasing BMI Z-score, as expected. However, we did not demonstrate higher leptin levels in autoantibody positive relatives, compared with autoantibody-negative control subjects. Thus, elevations in leptin levels do not appear to be a major determinant of whether an individual develops autoimmunity. If elevated BMI and associated elevations in leptin levels are risk factors for the initiation and/or progression of autoimmunity, they may act more as permissive factors in this process, in the setting of a certain degree of genetic predisposition. PMID- 16380227 TI - Comparative genomic and expression analysis of the conserved NTPDase gene family in Xenopus. AB - The purines, ATP and adenosine, are important signaling molecules in the nervous system. ATP is sequentially degraded to adenosine by the ectonucleotidase proteins. The NTPDase (or CD39) family is a subfamily of these enzymes, which consists of nine members in mammals. In Xenopus embryos, we have shown that ATP, and its antagonist adenosine, regulate the rundown of swimming and we therefore proposed that ectonucleotidase proteins are key regulators of locomotor activity. Here, we report the cloning of all nine members of the NTPDase family in Xenopus laevis and Xenopus tropicalis. Our phylogenetic analysis shows that this family is highly conserved between the frog species and also during vertebrate evolution. In the adult frog, NTPDase genes are broadly expressed. During development, all NTPDase genes, except for NTPDase8, are expressed and display a distinct specific expression pattern, suggesting potentially different functions of these proteins during embryogenesis of X. laevis. PMID- 16380229 TI - Type 2 autoimmune hepatitis murine model: the influence of genetic background in disease development. AB - Genetic predisposition is recognized as an important factor for the development of autoimmune hepatitis (AIH). To assess the potential contribution of MHC and non-MHC genes, type 2 AIH was reproduced in three mice strains, taking advantage of their different genetic makeup with regard to MHC and non-MHC genes. Mice (C57BL/6, 129/Sv and BALB/c) were DNA vaccinated with a pCMV-CTLA4-CYP2D6-FTCD plasmid coding for the extracellular region of CTLA-4 and for the antigenic region of the CYP2D6 and FTCD, and with pCMV-IL12. ALT and total IgG levels, liver histology, FACS analysis and liver T-cell cytotoxicity assays were monitored up to 8 months post-injection. C57BL/6 mice showed elevated serum ALT levels, autoantibodies, antigen-specific cytotoxic T-cells and lobular and periportal inflammatory infiltrate. The 129/Sv mice showed slightly elevated ALT levels, sparse liver lobular infiltrate and cytotoxic T-cells. The BALB/c mice showed no liver inflammation. All mice had elevated total serum IgG levels. This murine model of type 2 AIH shows that MHC and non-MHC genes contribute to the susceptibility to autoimmune hepatitis. The understanding of the genetic determinants implicated in AIH development will be a major advance in the study of its pathogenesis and could lead to a better diagnostic approach and preventive strategies. PMID- 16380230 TI - Thrombopoietin (TPO) induces c-myc expression through a PI3K- and MAPK-dependent pathway that is not mediated by Akt, PKCzeta or mTOR in TPO-dependent cell lines and primary megakaryocytes. AB - Thrombopoietin (TPO) and its receptor (c-Mpl) are the major regulators of megakaryocyte and platelet production and serve a critical and non-redundant role in hematopoietic stem cell (HSC) biology. TPO signals through the Jak-STAT, Ras Raf-MAPK, and PI3K pathways, and promotes survival, proliferation, and polyploidization in megakaryocytes. The proto-oncogene c-myc also plays an important role in many of these same processes. In this work we studied the regulated expression of c-myc in megakaryocytic cell lines and primary cells by quantitative real-time RT-PCR. We found that TPO induced expression of c-myc in 1 h in both hematopoietic cell lines (UT-7 and BaF3/Mpl) and mature murine megakaryocytes. The TPO-induced expression of c-myc was blocked by a phosphatidylinositol 3-kinase (PI3K) inhibitor, suggesting that TPO stimulated c myc expression through a PI3K-dependent pathway. Of interest, our study showed that overexpression of active Akt did not rescue the effect of PI3K blockade on c myc expression, rather, enhanced it. In addition, inhibitors of protein kinase C (PKC)zeta and the target of rapamycin (mTOR) also failed to affect c-myc mRNA expression, while c-myc mRNA expression was reduced by inhibition of the mitogen activated protein kinase (MAPK) pathway. Therefore, we conclude that TPO stimulates c-myc expression in primary megakaryocytes through a PI3K- and MAPK dependent pathway that is not mediated by Akt, PKCzeta or mTOR. PMID- 16380231 TI - Synergistic activation of PKD by the B cell antigen receptor and CD19 requires PI3K, Vav1 and PLCgamma. AB - Antigens coated with complement fragments coligate the B cell receptor (BCR) with the CD21/CD19 complex which results in synergistic activation of B cells. Previous studies identified PI3K, Vav proteins and PLCgamma as important components of this synergy. We now show that protein kinase D (also known as PKCmu) is also a point of convergence of these signalling pathways. We found that PKD activation upon BCR engagement or coligation of the BCR with CD19 is entirely dependent on PI3K and PLCgamma but differ in the requirement for Vav proteins. Whereas PKD activation is Vav1 and Vav2 dependent in response to BCR cross linking, PKD activation is sensitive to the lack of Vav1 under synergistic stimulation of BCR and CD19. These findings show that Vav proteins and PI3K regulation of PLCgamma contributes to the activation of PKD in response to BCR and or CD19 cross-linking. PMID- 16380232 TI - Epilepsy, surgery, and the elderly. AB - Treatment of elderly patients with epilepsy may present unique challenges to physicians. Co-morbid conditions and drugs to treat such conditions are common in elderly patients, possibly complicating epilepsy therapies that are dependent on drugs alone. For this reason, surgical intervention may be an attractive option for elderly patients with epilepsy, particularly for medically intractable patients with key disease features, such as lateralization and precisely localized epileptic foci. Curative procedures, including lobectomy and lesionectomy, are most likely to lead to seizure freedom, but not all patients are candidates for such procedures. When a curative surgical procedure is not an option, palliative procedures, including vagus nerve stimulation and deep brain stimulation, may be viable options. Vagus nerve stimulation has been reported to reduce seizure rates and improve quality of life in elderly patients with epilepsy. Currently, widespread therapeutic application of deep brain stimulation is limited by risks, costs, and pending studies. PMID- 16380233 TI - Influence of growth phase on the phospholipidic fatty acid composition of two marine bacterial strains in pure and mixed cultures. AB - This in vitro study was conducted in order to determine the effects of hydrocarbons and growth phase on the phospholipid ester-linked fatty acid composition of two marine sedimentary hydrocarbon-degrading bacteria. These two strains, namely Corynebacterium sp. and Sphingomonas sp. 2MPII, were cultivated on either a simple soluble substrate (ammonium acetate) or a hydrocarbon (respectively n-eicosane and phenanthrene). The incubations were stopped at different times corresponding to point of lag (2 days), exponential (7 days) and stationary phases (21 and 56 days). The effects of growth phase and hydrophobic substrates were successfully demonstrated by a simple index, given as the sum of saturated fatty acids divided by the sum of unsaturated fatty acids ( summation operatorSFA/ summation operatorMUFA), ranging from 1.4 to 3, 0.3 to 0.6, and 0.5 to 1.0 for Corynebacterium sp., Sphingomonas sp. 2MPII, and mixed cultures, respectively. This result was validated by a principal component analysis. In pure cultures, the phospholipid fatty acid (PLFA) composition was strongly influenced by both the carbon source and the growth phase. Nevertheless, the two strains showed different "behaviors". For 2MPII, the main PLFA composition changes were observed at 2 days while they were progressive as a function of time for Corynebacterium sp. These differences could explain the evolution of PLFAs of mixed cultures. PMID- 16380234 TI - A combination of direct viable count and fluorescent in situ hybridization for estimating Helicobacter pylori cell viability. AB - The viable but non-culturable (VBNC) stage of Helicobacter pylori may represent a problem of public health concern, since these cells cannot be detected by traditional culture methods. In this study, the direct viable count method (DVC) was modified and adapted to H. pylori analysis by testing different times of incubation and concentrations of DNA-gyrase inhibitors. The DVC procedure was combined with fluorescent in situ hybridization (FISH) for the specific detection of viable cells of H. pylori (DVC-FISH). Incubation with 0.5 microg/ml of novobiocin for 24 h provided the optimal conditions for obtaining 3-5 times the original size of Helicobacter viable cells. Field work performed with various types of water (freshwater and seawater) using the DVC-FISH approach enabled us to confirm the presence of VBNC H. pylori cells in 16 of the 45 analyzed samples. The combination of the modified DVC procedure with FISH can provide a rapid and specific method to detect and identify viable cells of H. pylori in environmental samples. PMID- 16380235 TI - The 2-year stability of neurological soft signs after a first episode of non affective psychosis. AB - We examined the 2-year stability of neurological soft signs (NSS) in 29 patients after a first episode of psychosis. The numbers of NSS at inclusion and at 2 years follow-up were similar, but there was a significant increase in the numbers of NSS in the sub-group of patients whose dosage of antipsychotic medication had increased over time. PMID- 16380237 TI - Meta-analysis of drop-out rates in randomised clinical trials, comparing typical and atypical antipsychotics in the treatment of schizophrenia. AB - OBJECTIVE: To assess antipsychotic medication in the treatment of schizophrenia, based on trial drop-out rates. METHOD: The studies included were randomised controlled trials that compared any of the four clinically best-established atypical antipsychotics (quetiapine, olanzapine, risperidone or clozapine) against either of two typical antipsychotics regarded as the gold standard (haloperidol or chlorpromazine). RESULTS: Meta-analysis indicated less risk of all-cause patient withdrawal from atypical medication trials where dosage was flexible, in both the short, relative risk (RR) 0.70 (95% CI 0.64-0.76), P<0.00001, and long term, RR 0.72 (0.65-0.80), P<0.00001. Similar results were observed for withdrawal due to adverse events, RR: 0.54 (0.41-0.72), P<0.0001. Nevertheless, the favourable effects of atypical medication disappeared in trials relying on fixed dosage. CONCLUSIONS: We detected a significant positive effect in terms of the outcome of treatment discontinuation for atypical versus typical medication, though only where the use of flexible rather than fixed doses (closer to an experimental control situation) was possible. PMID- 16380236 TI - Impact of cognitive-psychoeducational interventions in bipolar patients and their relatives. AB - BACKGROUND: In recent years, several controlled studies could show that psychoeducational interventions have been effective for relapse prevention in bipolar disorders. We therefore established a cognitive-psychoeducational group intervention with 14 sessions providing information about the illness, early warning signs, cognitive and behavioural strategies for stress management and social rhythm. Additionally we offered a group intervention for the patients' relatives. The objective of this study was to describe the outcome associated with our psychoeducational intervention in bipolar patients and their relatives. METHODS: Sixty-two bipolar patients attended 14 sessions (a 90 min) of cognitive psychoeducational group therapy. Patients' knowledge of bipolar disorder and their satisfaction with the treatment were assessed using self-developed questionnaires before and after the group intervention. Additionally, 49 relatives of bipolar patients received two psychoeducational workshops of 4 hours each. We assessed demographic variables, burden, high expressed emotion and depressive symptoms of the relatives before and after the two workshops and at 1 year follow-up. RESULTS: Patients significantly improved their knowledge of bipolar disorder. They also have benefited from the discussions and the exchange of useful coping strategies. Burden and high expressed emotions showed no significant reductions at post-assessment, however they were significantly reduced at 1-year follow-up. Relatives also felt significantly better informed about the illness. CONCLUSIONS: These findings show that psychoeducational interventions in bipolar patients and their relatives improve patients' and their relatives' knowledge of the illness and the burden of the disorder as well as high expressed emotions are reduced in relatives at 1-year follow-up. PMID- 16380238 TI - Imipramine treatment ameliorates corticosterone-induced alterations in the effects of 5-HT1A and 5-HT4 receptor activation in the CA1 area of rat hippocampus. AB - This study tested whether imipramine reverses adaptive modifications in the function of hippocampal 5-HT1A and 5-HT4 receptors induced by repetitive administration of corticosterone. Rats received corticosterone for 1 or 3 weeks or imipramine for 2 weeks. The fourth experimental group was treated with corticosterone for 3 weeks and additionally with imipramine, beginning on the eighth day of corticosterone administration. Hippocampal slices were prepared 48 h after the last drug administration. 5-HT1A and 5-HT4 receptor-mediated effects on CA1 population spike amplitude were measured. While repeated corticosterone attenuated the inhibitory effect of 5-HT1A receptor activation by 8-OH-DPAT and enhanced the excitatory effect of 5-HT4 receptor activation by zacopride, imipramine treatment of naive rats resulted in opposite changes. In the corticosterone plus imipramine group, the effect of 8-OH-DPAT and zacopride were not different from control, indicating that corticosterone-induced adaptive changes in the reactivity of 5-HT1A and 5-HT4 receptors were reversed by imipramine treatment. PMID- 16380240 TI - Chemokines and leukocyte trafficking in rheumatoid arthritis. AB - Leukocyte infiltration into the joint space and tissues is an essential component of the pathogenesis of rheumatoid arthritis (RA). In this review, we will summarize the current understanding of the mechanisms of leukocyte trafficking into the synovium, focusing on the role of adhesion molecules, chemokines, and chemokine receptors in synovial autoimmune inflammation. The process by which a circulating leukocyte decides to migrate into the synovium is highly regulated and involves the capture, firm adhesion, and transmigration of cells across the endothelial monolayer. Adhesion molecules and chemokine signals function in concert to mediate this process and to organize leukocytes into distinct structures within the synovium. Chemokines play a key regulatory role in organ specific leukocyte trafficking and activation by affecting integrin activation, chemotaxis, effector cell function, and cell survival. Consequently, chemokines, their receptors, and downstream signal transduction molecules are attractive therapeutic targets for RA. PMID- 16380239 TI - Does amygdalar perfusion correlate with antidepressant response to partial sleep deprivation in major depression? AB - This study used functional MRI (fMRI) to clarify the sites of brain activity associated with the antidepressant effects of sleep deprivation (SD). We hypothesized: (1) baseline perfusion in right and left amygdalae will be greater in responders than in nonresponders; (2) following partial sleep deprivation (PSD), perfusion in responders' right and left amygdalae would decrease. Seventeen unmedicated outpatients with current major depression and eight controls received perfusion-weighted fMRI and structural MRI at baseline and following 1 night of late-night PSD. Baseline bilateral amygdalar perfusion was greater in responders than nonresponders. Clusters involving both amygdalae decreased from baseline to PSD specifically in responders. Right amygdalar perfusion diverged with PSD, increasing in nonresponders and decreasing in responders. These novel amygdalar findings are consistent with the overarousal hypothesis of SD as well as other functional imaging studies showing increased baseline amygdalar activity in depression and decreased amygdalar activity with remission or antidepressant medications. PMID- 16380241 TI - Physicochemical characterization and drug release properties of Eudragit E PO/Eudragit L 100-55 interpolyelectrolyte complexes. AB - The formation of interpolyelectrolyte complexes (IPEC) between Eudragit E PO (EE) and Eudragit L 100-55 (EL) was investigated, using turbidimetry, apparent viscosity measurements, elementary analysis and MT-DSC. The structure of the synthesized IPEC was investigated using FT-IR spectroscopy. The binding ratio of a unit molecule of EL with EE was found to be approximately 1:1 at pH 5.5. Based on the results of elementary analysis and FT-IR, the binding ratio of each component in the solid complexes was very close to that observed in turbidity and apparent viscosity measurements and indicate that the synthesized products can be considered as IPEC. As a result of electrostatic interaction between the polymer chains, the glass transition temperature of the IPEC increased significantly. Due to the structure of the IPEC, two maxima were observed in the swelling behavior as a function of pH. The release of the model drug ibuprofen (IBF) was significantly retarded from tablets made up of the IPEC as compared with individual copolymers, its physical mixture and Eudragit RL PO (RL), RS PO (RS). PMID- 16380242 TI - TEWL measurements as a routine method for evaluating the integrity of epidermis sheets in static Franz type diffusion cells in vitro. Limitations shown by transport data testing. AB - The suitability of transepidermal water loss (TEWL) measurements in vitro as a barrier integrity test for human heat separated epidermis (HSE) was investigated. A model system consisting of a Teflon membrane mounted in Franz diffusion cells (FDC) filled with phosphate buffer saline (PBS) was set up. The membrane was used intact and punctured with a needle (up to five holes). After each puncturing the TEWL was measured. Only the TEWL of intact and punctured membrane differed significantly regardless of the number of holes. From three donors intact human HSE and punctured HSE were compared and no significant difference of the TEWL was found. Permeation experiments with flufenamic acid (FFA) showed a significantly higher diffusion rate through punctured HSE. TEWL and drug permeation were compared for skin stripped three, seven and 15 times prior to heat separation to an intact control group. Only the TEWL values of intact HSE and HSE stripped 15 times differed significantly. However, seven and 15 times stripping resulted in significantly higher diffusion rate. In conclusion, TEWL measurements can detect severe damage of the stratum corneum (SC) but not small changes, which nevertheless may already influence drug diffusion. Therefore, TEWL measurements appears to be of limited use as a barrier integrity test for human HSE in in vitro test systems. PMID- 16380243 TI - Preparation and in vitro evaluation of lipidic carriers and fillers for inhalation. AB - The present study relates to compositions of solid lipidic microparticles (SLmP), composed of biocompatible phospholipids and cholesterol, and their use as carriers or as fillers delivering drugs directly to the lungs via a dry powder inhaler (DPI). SLmP were obtained by spray-drying and were formulated as lipidic matrices entrapping budesonide or as physical blends (drug carrier). They were developed in order to improve the delivery of the active drug by the pulmonary route. The SLmP were evaluated for their physical characteristics and in vitro deposition measurements were performed using the Multi-stage Liquid Impinger (MsLI). The Pulmicort Turbuhaler DPI (AstraZeneca) was used as a comparator product. The SLmP appeared to be spherical low-density material characterized by a smooth surface. The mass median diameters (D(0.5)), and the volume mean diameters (D[4,3]) were tiny and ranged from 1.7 to 3.1 microm and from 2.0 to 3.9 microm, respectively. The SLmP formulations, delivered by the Cyclohaler inhaler, were found to emit a fine particle dose (FPD) of 93-113 microg, which is very promising comparing to the FPD (68 microg) delivered by the Pulmicort Turbuhaler. PMID- 16380244 TI - Starch metabolism in Leucoagaricus gongylophorus, the symbiotic fungus of leaf cutting ants. AB - Leucoagaricus gongylophorus, the symbiotic fungus of the leaf-cutting ants, degrades starch, this degradation being supposed to occur in the plant material which leafcutters forage to the nests, generating most of the glucose which the ants utilize for food. In the present investigation, we show that laboratory cultures of L. gongylophorus produce extracellular alpha-amylase and maltase which degrade starch to glucose, reinforcing that the ants can obtain glucose from starch through the symbiotic fungus. Glucose was found to repress alpha amylase and, more severely, maltase activity, thus repressing starch degradation by L. gongylophorus, so that we hypothesize that: (1) glucose down-regulation of starch degradation also occurs in the Atta sexdens fungus garden; (2) glucose consumption from the fungus garden by A. sexdens stimulates degradation of starch from plant material by L. gongylophorus, which may represent a mechanism by which leafcutters can control enzyme production by the symbiotic fungus. Since glucose is found in the fungus garden inside the nests, down-regulation of starch degradation by glucose is supposed to occur in the nest and play a part in the control of fungal enzyme production by leafcutters. PMID- 16380245 TI - Inhibition of hyaluronan hydrolysis catalysed by hyaluronidase at high substrate concentration and low ionic strength. AB - Hyaluronidase and high levels of hyaluronan are found together in tumours. It is highly likely that hyaluronidase activity controls the balance between high molecular mass hyaluronan and oligosaccharides, and thus plays an important role in cancer development. The hyaluronan hydrolysis catalysed by bovine testicular hyaluronidase was studied as a model. The kinetics was investigated at pH 5 and 37 degrees C using the colorimetric N-acetyl-d-glucosamine reducing end assay method. While the substrate dependence obtained in the presence of 0.15 mol L(-1) ionic strength exhibited a Michaelis-Menten behaviour, an atypical behaviour was observed under low ionic strength: for increasing hyaluronan concentrations, the initial reaction rate increased, reached a maximum and then decreased to a very low level, close to zero at high substrate concentrations. One of the various hypotheses examined to explain this atypical behaviour is the formation of non specific complexes between hyaluronan and hyaluronidase based on electrostatic interactions. This hypothesis is the only one that can explain all the experimental results including the variation of the reaction medium turbidity as a function of time and the influence on the initial reaction rate of the hyaluronan concentration over hyaluronidase concentration. However, phenomena such as the high viscosity of highly concentrated hyaluronan solutions or the steric exclusion of hyaluronidase from hyaluronan solutions may contribute to the atypical behaviour. Finally, the biological implications of the non-linear and non-monotonous shape of the hyaluronan-hyaluronidase substrate dependence in the regulation of the hyaluronan chain molecular mass are discussed, in particular in the case of cancer development. PMID- 16380246 TI - Preferential production of prostaglandin D2 by lipopolysaccharide stimulated human choriodecidual explants. AB - It has been postulated that the progression of a pregnancy to term is, in part, the result of a relative maternal Th2 immunological state. Prostaglandins (PG) are critical mediators throughout pregnancy. Recent studies have demonstrated that one PG, PGD2, may be a mediator of a Th2 immunological state. To date, very little is known about the factors that regulate of PGD2 production by human gestational tissues. Placentae were collected from women undergoing Caesarean sections at term. Amnion was separated from the choriodecidua and choriodecidual explants established. Explants were allowed to equilibrate overnight in media containing 10% fetal calf serum. The following day, media were replaced with serum free media and then after an additional 24-h, media were collected and the wet weight of the tissues determined. Production rates of PGs were determined using radioimmunoassays. At all concentrations tested, LPS significantly enhanced PGD2 production by human choriodecidual explants compared to PGE2 and PGF2alpha production. Neutralization of TNF-alpha and IL-10 further increased the production of LPS stimulated PGD2 production. We suggest that a novel stimulatory pathway that drives the production of PGD2 has been uncovered. PMID- 16380247 TI - Application of a gas-solid fluidized bed separator for shredded municipal bulky solid waste separation. AB - A laboratory-scale gas-solid fluidized bed separator able to separate fractions of 5.6-50mm was used for separation of shredded municipal bulky waste (SBW) into combustibles and incombustibles. In batch-scale tests, it was found that accumulation of SBW in the bottom of the bed significantly reduced the separation efficiency. In this study, stirring was shown to be effective in preventing this accumulation. Flexible sheet materials such as paper and film plastics also significantly decreased the separation efficiency. In batch-scale tests, an overall efficiency of 90% was obtained when flexible materials such as film plastics and paper were excluded from the feed SBW. In continuous feeding tests, purities of the float and sink fractions attained 95% and 86% efficiencies, respectively, with an overall efficiency of 79%. The effect of feedstock shape on separation efficiency was also investigated. This study revealed that large particles can be properly separated on the basis of density, while the shape of the material significantly influenced behavior in the fluidizing bed. PMID- 16380248 TI - Ets2 transcription factor in normal and neoplastic human breast tissue. AB - The Ets family of transcription factors regulate the expression of multiple genes involved in tumour formation and progression. The aim of this work was to test the hypothesis that the expression of Ets2 in breast cancers was associated with parameters of tumour progression and metastasis. Using reverse-transcriptase polymerase chain reaction (RT-PCR), Ets2 mRNA was detected in 69% of 181 breast carcinomas, 63% of 43 fibroadenomas and 47% of 43 specimens of normal breast tissue. Levels were significantly higher in carcinomas compared with normal breast tissue (P = 0.006). Using Western blotting, Ets2 protein was found to migrate as two bands with molecular masses of 52 kDa (p52) and 54kDa (p54). Levels of both proteins were significantly higher in the carcinomas compared with both fibroadenomas (P = 0.0001) and normal breast tissue (P = 0.0001). In the carcinomas, a significant relationship was found between the p52 and p54 form of Ets2 (r = 0.51, P < 0.0001; Spearman correlation). Also, in the carcinomas, a significant correlation was found between both forms of Ets2 protein and urokinase plasminogen activator (uPA) (for p52, r = 0.43, P = 0.0005, n = 68; for p54, r = 0.50, P = 0.0001, n = 68). As Ets2 binding sites are present on the uPA promoter, Ets2 may be one of the transcription factors regulating uPA expression in human breast cancer. PMID- 16380249 TI - Preparation and characterization of activated carbon from palm shell by chemical activation with K2CO3. AB - Palm shell was used to prepare activated carbon using potassium carbonate (K2CO3) as activating agent. The influence of carbonization temperatures (600-1000 degrees C) and impregnation ratios (0.5-2.0) of the prepared activated carbon on the pore development and yield were investigated. Results showed that in all cases, increasing the carbonization temperature and impregnation ratio, the yield decreased, while the adsorption of CO2 increased, progressively. Specific surface area of activated carbon was maximum about 1170 m2/g at 800 degrees C with activation duration of 2 h and at an impregnation ratio of 1.0. PMID- 16380250 TI - 4-Aminophenoxyacetic acids as a novel class of reversible cathepsin K inhibitors. AB - We have designed and synthesized a novel series of 3-biphenylamino acid amides as cathepsin K inhibitors based on compound I. In these inhibitors, we have discovered 4-aminophenoxyacetic acids 43 and 47 with good IC(50) values, although lipophilic groups are favorable for the hydrophobic S1' pocket. PMID- 16380251 TI - Discovery and synthesis of a novel series of quinoline-based thrombin receptor (PAR-1) antagonists. AB - The design, synthesis, and SAR studies of a structurally novel series of highly potent thrombin receptor (PAR-1) antagonists are described. Compound 30 is a highly potent thrombin receptor antagonist (IC(50)=6.3 nM), a related compound 36 showing efficacy in a monkey ex vivo study. PMID- 16380252 TI - 5-Heteroatom-substituted pyrazoles as canine COX-2 inhibitors: Part 2. Structure activity relationship studies of 5-alkylethers and 5-thioethers. AB - Structure-activity relationship (SAR) studies of novel 2-[3-trifluoromethyl-5 alkyl(thio)ether pyrazo-1-yl]-5-methanesulfonyl pyridine derivatives for canine COX enzymes are described. The 4-cyano-5-alkyl ethers were found to have excellent potency and selectivity, whereas the 5-thioethers were potent but less selective than the ether analogs in a canine whole blood (CWB) COX-2 assay. PMID- 16380253 TI - Antiangiogenic activity of deoxoartemisinin derivatives on chorioallantoic membrane. AB - Non acetal-type derivatives at C-12 of artemisinin and their novel dimers including a fullerene conjugate were synthesized and some of them showed potent in vivo antiangiogenic activity on chorioallantoic membrane higher than or comparable to those of fumagillin and thalidomide. PMID- 16380254 TI - Synthesis and biological evaluation of 5-[(aryl)(1H-imidazol-1-yl)methyl]-1H indoles: potent and selective aromatase inhibitors. AB - The synthesis and the aromatase (CYP19) inhibitory activity of 5-[(aryl)(imidazol 1-yl)methyl]-1H-indoles were reported. Among the tested racemate compounds, 5-[(4 chlorophenyl)(1H-imidazol-1-yl)methyl]-1H-indole 8b emerged as a potent CYP19 inhibitor (IC(50)=15.3 nM). Chiral chromatography allowed isolation of the (+) enantiomer 8b2, which was about twice as active as the racemate (IC(50)=9 nM). PMID- 16380255 TI - Phenyl ureas of creatinine as mGluR5 antagonists. A structure-activity relationship study of fenobam analogues. AB - Fenobam (1) was developed by McNeil Laboratories as an anxiolytic agent with an unknown molecular target in the late 1970s. In a recent publication, it was revealed that fenobam is a non-competitive mGluR5 antagonist. Herein, we present the structure-activity relationship of fenobam and its analogues and similarities between the SAR of mGluR5 antagonism and the SAR of CNS properties originally reported by McNeil are discussed. PMID- 16380256 TI - Indole-glucosides as novel sodium glucose co-transporter 2 (SGLT2) inhibitors. Part 2. AB - A series of indole-O-glucosides and C-glucosides was synthesized and evaluated in SGLT1 and SGLT2 cell-based functional assays. Compounds 2a and 2o were identified as potent SGLT2 inhibitors and screened in ZDF rats. PMID- 16380257 TI - Structure-activity relationships of alpha-amino acid ligands for the alpha2delta subunit of voltage-gated calcium channels. AB - A series of alpha-amino acids were identified as ligands which compete with gabapentin for binding to the alpha(2)delta subunit of voltage-dependent Ca(2+) channels. Potent analogs were identified. Their activity in an in vivo pain assay is described. PMID- 16380258 TI - Novel dual inhibitors of calpain and lipid peroxidation with enhanced cellular activity. AB - A series of dipeptides with dual inhibitory activities on calpain and lipid peroxidation were prepared to target the intracellular calpain. This optimization program focused on the variations of the linker and the N-terminal amino acid of the peptidic core. Two compounds 6d-05 and 6d-08 exhibited potent intracellular calpain inhibition. The polar surface area and the number of rotors appeared to be critical descriptors to account for the behavior of these hybrid molecules in the cellular calpain assay. PMID- 16380259 TI - Inhibitors of epidermal growth factor receptor tyrosine kinase: Novel C-5 substituted anilinoquinazolines designed to target the ribose pocket. AB - A series of novel C-5 substituted anilinoquinazolines, selected on the basis of docking experiments and overlays with ATP in the active site of EGFR tyrosine kinase, have been prepared and found to be potent inhibitors. In vivo pharmacokinetics and disease model activity are discussed. PMID- 16380260 TI - Parting company with the chromosome segregation and aneuploidy series. PMID- 16380261 TI - Fluorescein-based amino acids for solid phase synthesis of fluorogenic protease substrates. AB - An efficient synthesis of new type fluorescent amino acids is described. The Fmoc protected dyes can be prepared in a four-step procedure with approximately 30% overall yield from aminofluoresceins and other inexpensive commercially available precursors. The dyes are much more photostable compared to fluorescein and exhibit constant pH-independent fluorescence that is advantageous in biological applications. The Fmoc-protected fluorescent amino acids are ready for use in solid phase peptide synthesis. As a proof of concept, a fluorogenic papain substrate was synthesized and employed for on-bead detection of the protease activity. By using a novel technique for quantitative analysis of bead fluorescence, a approximately 2.7-fold increase in mean bead brightness was measured and was attributed to substrate cleavage by papain. The new type fluorescent amino acids seem to be a promising tool for the synthesis of fluorescent peptide ligands and fluorogenic protease substrates. PMID- 16380262 TI - Half-life of 176Lu. AB - The half-life of 176Lu, measured using gamma-ray spectrometry, has been found to be (3.56+/-0.07)x10(10)yr. Comparison has been made between present results and literature data. PMID- 16380263 TI - Determination of 243Am by pulse shape discrimination liquid scintillation spectrometry. AB - Alpha specific activity of 243Am was determined using pulse shape discrimination in liquid scintillation spectrometry. 238Pu, 36Cl and 239Np (purified from 243Am) were used for obtaining the spillover of alpha/beta particles into the beta/alpha channels, respectively. Synthetic mixtures of 241Am/243Am were prepared. Using the alpha-specific activity, weights of the stock solutions used and the half life of 241Am and 243Am isotopes, the expected 241Am/243Am atom ratios in the mixtures were determined and compared with those obtained by thermal ionization mass spectrometry (TIMS). An agreement of about 1% was obtained between the 241Am/243Am atom ratios determined by the two methods. This shows that liquid scintillation counting with pulse shape discrimination can be used for 243Am determination with an accuracy better than 1%. PMID- 16380264 TI - Reduced ubiquitin C-terminal hydrolase-1 expression levels in dementia with Lewy bodies. AB - Parkinson disease (PD) and dementia with Lewy bodies (DLB) are characterized by the accumulation of abnormal alpha-synuclein and ubiquitin in protein aggregates conforming Lewy bodies and Lewy neurites. Ubiquitin C-terminal hydrolase-1 (UCHL 1) disassembles polyubiquitin chains to increase the availability of free monomeric ubiquitin to the ubiquitin proteasome system (UPS) thus favoring protein degradation. Since mutations in the UCHL-1 gene, reducing UPS activity by 50%, have been reported in autosomal dominant PD, and UCHL-1 inhibition results in the formation of alpha-synuclein aggregates in mesencephalic cultured neurons, the present study was initiated to test UCHL-1 mRNA and protein levels in post mortem frontal cortex (area 8) of PD and DLB cases, compared with age-matched controls. TaqMan PCR assays, and Western blots demonstrated down-regulation of UCHL-1 mRNA and UCHL-1 protein in the cerebral cortex in DLB (either in pure forms, not associated with Alzheimer disease: AD, and in common forms, with accompanying AD changes), but not in PD, when compared with age-matched controls. Interestingly, UCHL-1 mRNA and protein expressions were reduced in the medulla oblongata in the same PD cases. Moreover, UCHL-1 protein was decreased in the substantia nigra in cases with Lewy body pathology. UCHL-1 down-regulation was not associated with reduced protein levels of several proteasomal subunits, including 20SX, 20SY, 19S and 11Salpha. Yet UCHL-3 expression was reduced in the cerebral cortex of PD and DLB patients. Together, these observations show reduced UCHL-1 expression as a contributory factor in the abnormal protein aggregation in DLB, and points UCHL-1 as a putative therapeutic target in the treatment of DLB. PMID- 16380265 TI - GDNF is a chemoattractant factor for neuronal precursor cells in the rostral migratory stream. AB - Olfactory bulb (OB) interneurons are generated from neuroblast cells derived from the anterior subventricular zone (SVZa) of the forebrain. The mechanisms guiding the rostral migration of these neuronal precursors are not well understood. Here, we show that glial cell line-derived neurotrophic factor (GDNF) is produced in the olfactory bulb but distributed along the rostral migratory stream (RMS) in a pattern concordant with the expression of its GPI-anchored receptor GFRalpha1. We demonstrate that GDNF is a chemoattractant factor for RMS-derived neuronal precursors, but not for SVZa neuroblast cells. In agreement with this, GDNF increased Cyclin-dependent kinase 5 (Cdk5) activity in RMS cells, a kinase critically involved in neuronal migration and guidance. GDNF-mediated cell chemoattraction was abrogated in RMS explants treated with the Cdk5 inhibitor Roscovitine as well as in RMS explants isolated from Ncam mutant mice. Chemical cross-linking assays showed that 125I-GDNF is able to interact directly with NCAM in RMS-derived cells. Taken together, these data demonstrate that GDNF is a direct chemoattractant factor for neuroblast cells migrating along the RMS and support the participation of NCAM during this guidance process. PMID- 16380266 TI - Over-expression, purification, and characterization of aminopeptidase N from Escherichia coli. AB - The gene from Escherichia coli encoding aminopeptidase N (PepN) was subcloned into pET-26b, and PepN was over-expressed in BL21(DE3) E. coli and purified using Q-Sepharose chromatography. This protocol yielded over 17 mg of purified, recombinant PepN per liter of growth culture under optimum conditions. Gel filtration chromatography revealed that recombinant PepN exists as a monomer. MALDI-TOF mass spectra showed that the enzyme has a molecular mass of 98,750 Da, and steady-state kinetic studies revealed that as-isolated, recombinant PepN exhibits a k(cat) of 354 +/- 11s(-1) and a K(m) of 376 +/- 39 microM when using L alanine-p-nitroanilide as the substrate. Metal analyses demonstrated that as isolated, recombinant PepN binds 0.5 and <0.1 equivalents of iron and zinc, respectively. The addition of Zn(II) to recombinant PepN inhibits catalytic activity, while the addition of iron causes a slight decrease or no change in activity. Further metal binding studies revealed that recombinant PepN tightly binds 5 equivalents of iron and <0.1 equivalents of Zn(II). By using this over expression and purification system, E. coli PepN can now be obtained in quantities necessary for structural characterization and possibly inhibitor design efforts. PMID- 16380267 TI - Expression, purification, and characterization of His-tagged Staphylococcus xylosus lipase wild-type and its mutant Asp 290 Ala. AB - The gene encoding the extracellular lipase of Staphylococcus xylosus (SXL) was cloned using PCR technique. The sequence corresponding to the mature lipase was subcloned in the pET-14b expression vector, with a strong T7 promoter, to construct a recombinant lipase protein containing six histidine residues at the N terminal. High level expression of the lipase by Escherichia coli BL21 (DE3) cells harbouring the lipase gene containing expression vector was observed upon induction with 0.4 mM IPTG at 37 degrees C. One-step purification of the recombinant lipase was achieved with Ni-NTA resin. The specific activity of the purified His-tagged SXL was 1500 or 850 U/mg using tributyrin or olive oil emulsion as substrate, respectively. It has been proposed that the region near the residue Asp290 could be involved in the selection of the substrate. Therefore, we also mutated the residue Asp 290 by Ala using site-directed mutagenesis. The mutant SXL-D290A was overexpressed in E. coli BL21 (DE3) and purified with the same nickel metal affinity column. The specific activity of the purified His-tagged SXL-D290A mutant was 1000 U/mg using either tributyrin or olive oil emulsion as substrate. A comparative study of the wild type (His(6) SXL) and the mutant (His(6)-SXL-D290A) proteins was carried out. Our results confirmed that Asp290 is important for the chain length specificity and catalytic efficiency of the enzyme. PMID- 16380268 TI - Purification and cytotoxic properties of Bacillus cereus hemolysin II. AB - The hemolysin II from Bacillus cereus, HlyII, is a member of the beta-barrel pore forming toxin family of secreted microbial proteins that includes the Staphylococcus aureus alpha-toxin. Compared with other proteins of the family, hemolysin II has 90 extra amino acids at its C-terminus. To examine more closely the cytotoxic and pore-forming properties of the protein, we have cloned and expressed it in Escherichia coli. We developed a purification procedure for the matured HlyII protein from both culture media and cell extracts using a combination of cation exchange and affinity chromatography together with gel filtration. In both cases, the fully processed HlyII protein was purified as confirmed by N-terminal sequence analysis. The HlyII protein exhibits cytolytic activity of different extent on erythrocytes from various kinds of mammals. The results presented here show for the first time that two types of human cells are sensitive to HlyII action. In view of its broad cytotoxic activity as well as the ability to interact with artificial membranes, we assume that HlyII needs no specific receptor to bind to cell membranes. PMID- 16380269 TI - Cloning, expression, and purification of the general stress protein YhbO from Escherichia coli. AB - We cloned, expressed, and purified the Escherichia coli yhbO gene product, which is an amino acid sequence homolog to the Bacillus subtilis general stress protein 18 (the yfkM gene product), the Pyrococcus furiosus intracellular protease PfpI, and the human Parkinson disease protein DJ-1. The gene coding for YhbO was generated by amplifying the yhbO gene from E. coli by polymerase chain reaction. It was inserted into the expression plasmid pET-21a, under the transcriptional control of the bacteriophage T7 promoter and lac operator. A BL21 (DE3) E. coli strain transformed with the YhbO-expression vector, pET-21a-yhbO, accumulates large amounts of a soluble protein with a molecular mass of 20 kDa in SDS-PAGE that matches the expected YhbO molecular weight. YhbO was purified to homogeneity by ion exchange chromatography and hydroxyapatite chromatography, and its identity was confirmed by N-terminal sequencing and mass spectrometry analysis. The native protein exists in monomeric, trimeric, and hexameric forms. We also report a strong sequence homology between YhbO and the general stress protein YfkM (64% identities), which suggests that YhbO is a stress protein, and a strong structural homology between YhbO and the Pyrococcus horikoshii intracellular protease PhpI. We could not, however, detect any proteolytic or peptidolytic activity of YhbO, using classical biochemical substrates. PMID- 16380270 TI - Shared networks for auditory and motor processing in professional pianists: evidence from fMRI conjunction. AB - To investigate cortical auditory and motor coupling in professional musicians, we compared the functional magnetic resonance imaging (fMRI) activity of seven pianists to seven non-musicians utilizing a passive task paradigm established in a previous learning study. The tasks involved either passively listening to short piano melodies or pressing keys on a mute MRI-compliant piano keyboard. Both groups were matched with respect to age and gender, and did not exhibit any overt performance differences in the keypressing task. The professional pianists showed increased activity compared to the non-musicians in a distributed cortical network during both the acoustic and the mute motion-related task. A conjunction analysis revealed a distinct musicianship-specific network being co-activated during either task type, indicating areas involved in auditory-sensorimotor integration. This network is comprised of dorsolateral and inferior frontal cortex (including Broca's area), the superior temporal gyrus (Wernicke's area), the supramarginal gyrus, and supplementary motor and premotor areas. PMID- 16380271 TI - Self-initiated movements in chronic prefrontal traumatic brain injury: an event related functional MRI study. AB - Electrophysiological studies found traumatic lesions of the prefrontal cortex (PFC) to affect the preparation of self-initiated movements. However, a precise localization of the observed effects has not been reported yet. Moreover, previous reports did not account for effects of lesion location. Therefore, the present study utilized functional MRI to investigate the influence of predominantly left or right hemispheric PFC lesions on movement preparation in chronic traumatic brain injury (TBI) patients. 18 TBI patients with MRI-defined contusions of the PFC as well as 18 healthy control subjects matched for age and gender were examined. Patients were divided into subgroups of predominantly left or right hemispheric lesions. During functional image acquisition, subjects performed self-initiated abductions of their right index finger. Neuropsychological tests of attention and working memory, which are supposed to modulate preparatory processes, were conducted. Patients with predominantly left contusions demonstrated enhanced activity of the dorsal lateral premotor cortex in comparison to healthy control subjects. In predominantly right lesioned patients, reduced activity within the right caudate head was observed. Compared to predominantly left lesioned patients, neuropsychological tests yielded reduced task performances in the right lesion subgroup. In line with previous electrophysiological research, imaging results of the present study are interpreted to represent altered preparatory networks in chronic prefrontal TBI patients. Since attentional and working memory functions are supposed to modulate preparatory processes, differences between the patient subgroups are suggested to result from the more pronounced cognitive impairments in the right-lesioned group. PMID- 16380272 TI - Parieto-occipital grey matter abnormalities in children with Williams syndrome. AB - Williams syndrome (WS) is a neurodevelopmental disorder resulting from a hemizygous deletion of chromosome 7q11.23. The phenotype of WS consists of typical dysmorphic features, supravalvular aortic stenosis, infantile hypercalcemia and growth retardation. While language and facial recognition seem to be relatively spared, visuospatial constructive disabilities are a hallmark of the neurobehavioral profile of WS. In order to search for actual structural abnormalities underlying this precisely defined neurodevelopmental disorder, we performed anatomical magnetic resonance imaging (MRI) in 9 WS children (11.6 +/- 3.1 years; age range: 5.5-15 years) and 11 normal age-matched control children (11.8 +/- 2.2 years; age range: 8-15 years) using voxel-based morphometry (VBM). VBM is a fully automated whole-brain technique that delivers a voxel-wise assessment of regional grey and white matter concentration. A significant decrease in grey matter concentration was detected in the left parieto-occipital region of WS children (P < 0.05 corrected height threshold). The location of this abnormality in WS children coincides with the location of the structural abnormality previously described using the same method in 13 WS adults. These parieto-occipital abnormalities are consistent with the cognitive profile of WS which includes severe visuospatial construction and numerical cognition deficits. The demonstration of identical structural abnormalities in both adults and children argues for their early origin. Additionally, our study provides support for the use of advanced structural imaging techniques in children, in order to improve our understanding of neurobehavioral phenotypes associated with well defined genetic disorders. PMID- 16380273 TI - Common MRI acquisition non-idealities significantly impact the output of the boundary shift integral method of measuring brain atrophy on serial MRI. AB - Measuring rates of brain atrophy from serial magnetic resonance imaging (MRI) studies is an attractive way to assess disease progression in neurodegenerative disorders, particularly Alzheimer's disease (AD). A widely recognized approach is the boundary shift integral (BSI). The objective of this study was to evaluate how several common scan non-idealities affect the output of the BSI algorithm. We created three types of image non-idealities between the image volumes in a serial pair used to measure between-scan change: inconsistent image contrast between serial scans, head motion, and poor signal-to-noise (SNR). In theory the BSI volume difference measured between each pair of images should be zero and any deviation from zero should represent corruption of the BSI measurement by some non-ideality intentionally introduced into the second scan in the pair. Two different BSI measures were evaluated, whole brain and ventricle. As the severity of motion, noise, and non-congruent image contrast increased in the second scan, the calculated BSI values deviated progressively more from the expected value of zero. This study illustrates the magnitude of the error in measures of change in brain and ventricle volume across serial MRI scans that can result from commonly encountered deviations from ideal image quality. The magnitudes of some of the measurement errors seen in this study exceed the disease effect in AD shown in various publications, which range from 1% to 2.78% per year for whole brain atrophy and 5.4% to 13.8% per year for ventricle expansion (Table 1). For example, measurement error may exceed 100% if image contrast properties dramatically differ between the two scans in a measurement pair. Methods to maximize consistency of image quality over time are an essential component of any quantitative serial MRI study. PMID- 16380274 TI - Electrocardiogram, hemodynamics, and core body temperatures of the normal freely moving laboratory beagle dog by remote radiotelemetry. AB - INTRODUCTION: The objectives of this study were to provide baseline normative values for circadian changes in the time-series data collected over the course of a normal day in laboratory-housed dogs and to assess the relative efficiency of standard correction formulas to correct for the variations in QT intervals and heart rate functions. METHODS: One hundred and twenty-three beagle dogs (65 M, 58 F) were equipped with radiotelemetry transmitters and continuously monitored, while freely moving in their home cages. Electrocardiograms (ECGs), hemodynamic parameters (diastolic, systolic, and mean arterial pressures) as well as core body temperatures were recorded for 22 h. RESULTS AND DISCUSSION: Blood pressures and core body temperatures demonstrated only very slight variations in their respective values over the 22-h monitoring period. ECGs were measured by a computerized waveform analysis program and quantitative elements reported as RR, PR, QRS, and QT intervals. Little circadian rhythmicity was demonstrated in the ECG intervals. Standard study-specific correction formulas appeared to satisfactorily normalize (i.e., compensate for) the relationship between heart rate and QT intervals in these beagle dogs but elevated the values of the QTc as compared to the uncorrected QT intervals. In sharp contrast, a subject-specific correction method based on analysis of covariance produced a more linear function between heart rates and QT intervals and, more importantly, provided QTc values within the normal range of actual, recorded QT interval data. PMID- 16380275 TI - Ferroportin is a monomer in vivo in mice. AB - Ferroportin (FPN) is the main iron export protein in mammals. The actual structure of FPN in vivo and the pathogenesis of ferroportin-related disease are unknown. We aimed at studying the structure and biochemical properties of FPN in mouse tissues that are key for iron homeostasis during various iron manipulations in vivo. We performed glycosylation and oligomerization studies in spleen and liver extracts from mice fed a standard, iron-deprived or iron-enriched diet for 5 months. Purification by affinity chromatography and sucrose gradient show that FPN is not part of a large multiprotein complex. Dietary manipulations did not affect the monomeric status of the native or denatured protein. The glycosylation studies showed that ferroportin is digested by peptide: N-glycosidase F but not by endoglycosidase H. The same results were obtained using protein extracts from iron-deficient or iron-loaded mice. In conclusion, our studies indicate that mouse FPN, regardless of the tissue iron status, is glycosylated but not enriched in mannose residues, and that exists mainly in monomeric form. The latter finding may have important implications for understanding the pathogenesis of the disease due to ferroportin mutations. PMID- 16380276 TI - Secondary insomnia: a myth dismissed. PMID- 16380277 TI - Computational and comparative analyses of 150 full-length cDNA sequences from the oomycete plant pathogen Phytophthora infestans. AB - Phytophthora infestans is a devastating phytopathogenic oomycete that causes late blight on tomato and potato. Recent genome sequencing efforts of P. infestans and other Phytophthora species are generating vast amounts of sequence data providing opportunities to unlock the complex nature of pathogenesis. However, accurate annotation of Phytophthora genomes will be a significant challenge. Most of the information about gene structure in these species was gathered from a handful of genes resulting in significant limitations for development of ab initio gene calling programs. In this study, we collected a total of 150 bioinformatically determined near full-length cDNA (FLcDNA) sequences of P. infestans that were predicted to contain full open reading frame sequences. We performed detailed computational analyses of these FLcDNA sequences to obtain a snapshot of P. infestans gene structure, gauge the degree of sequence conservation between P. infestans genes and those of Phytophthora sojae and Phytophthora ramorum, and identify patterns of gene conservation between P. infestans and various eukaryotes, particularly fungi, for which genome-wide translated protein sequences are available. These analyses helped us to define the structural characteristics of P. infestans genes using a validated data set. We also determined the degree of sequence conservation within the genus Phytophthora and identified a set of fast evolving genes. Finally, we identified a set of genes that are shared between Phytophthora and fungal phytopathogens but absent in animal fungal pathogens. These results confirm that plant pathogenic oomycetes and fungi share virulence components, and suggest that eukaryotic microbial pathogens that share similar lifestyles also share a similar set of genes independently of their phylogenetic relatedness. PMID- 16380278 TI - Tissue astaxanthin and canthaxanthin distribution in rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar). AB - A comparative investigation of tissue carotenoid distribution between rainbow trout, Oncorhynchus mykiss, and Atlantic salmon, Salmo salar, was undertaken to identify the relative efficiency of utilization of astaxanthin and canthaxanthin. Higher apparent digestibility coefficients (ADCs) (96% in trout vs. 28-31% in salmon; P<0.05), and pigment retention efficiencies (11.5-12.5% in trout vs. 5.5% in salmon; P<0.05), for both astaxanthin and canthaxanthin, were observed for rainbow trout. Astaxanthin deposition was higher than canthaxanthin in rainbow trout, while the reverse was true for Atlantic salmon, suggesting species specificity in carotenoid utilization. The white muscle (95% in trout vs. 93% in salmon) and kidneys (0.5% in trout vs. 0.2% in salmon) represented higher proportions of the total body carotenoid pool in rainbow trout than in Atlantic salmon (P<0.05), whereas the liver was a more important storage organ in Atlantic salmon (2-6% in salmon vs. 0.2% in trout; P<0.05). The liver and kidney appeared to be important sites of carotenoid catabolism based on the relative proportion of the peak chromatogram of the fed carotenoid in both species, with the pyloric caecae and hind gut being more important in Atlantic salmon than in the rainbow trout. Liver catabolism is suspected to be a critical determinant in carotenoid clearance, with higher catabolism expected in Atlantic salmon than in rainbow trout. PMID- 16380279 TI - Respiratory allocation and standard rate of metabolism in the African lungfish, Protopterus aethiopicus. AB - This paper quantifies the relationship between respiratory allocation (air vs. water) and the standard rate of metabolism (SMR) in the primitive air-breathing lungfish, Protopterus aethiopicus. Simultaneous measurements of oxygen consumed from both air and water were made to determine the SMR at ecologically relevant aquatic oxygen levels for juveniles 2 to 221 g. Total metabolic rate was positively correlated with body mass with a scaling exponent of 0.78. Aerial oxygen consumption averaged 98% (range=94% to 100%) of total respiratory allocation under low aquatic oxygen levels. Measurements of oxygen consumption made across a gradient of dissolved oxygen from normoxia to anoxia showed that P. aethiopicus maintains its SMR despite a change in respiratory allocation between water and air. PMID- 16380280 TI - Ontogenetic shifts in thermal tolerance, selected body temperature and thermal dependence of food assimilation and locomotor performance in a lacertid lizard, Eremias brenchleyi. AB - We used Eremias brenchleyi as a model animal to examine differences in thermal tolerance, selected body temperature, and the thermal dependence of food assimilation and locomotor performance between juvenile and adult lizards. Adults selected higher body temperatures (33.5 vs. 31.7 degrees C) and were able to tolerate a wider range of body temperatures (3.4-43.6 vs. 5.1-40.8 degrees C) than juveniles. Within the body temperature range of 26-38 degrees C, adults overall ate more than juveniles, and food passage rate was faster in adults than juveniles. Apparent digestive coefficient (ADC) and assimilation efficiency (AE) varied among temperature treatments but no clear temperature associated patterns could be discerned for these two variables. At each test temperature ADC and AE were both higher in adults than in juveniles. Sprint speed increased with increase in body temperature at lower body temperatures, but decreased at higher body temperatures. At each test temperature adults ran faster than did juveniles, and the range of body temperatures where lizards maintained 90% of maximum speed differed between adults (27-34 degrees C) and juveniles (29-37 degrees C). Optimal temperatures and thermal sensitivities differed between food assimilation and sprint speed. Our results not only show strong patterns of ontogenetic variation in thermal tolerance, selected body temperature and thermal dependence of food assimilation and locomotor performance in E. brenchleyi, but also add support for the multiple optima hypothesis for the thermal dependence of behavioral and physiological variables in reptiles. PMID- 16380281 TI - Purification and partial characterization of two acid phosphatase forms from pearl oyster (Pinctada fucata). AB - The present study describes the details about the acid phosphatase forms in the pearl oyster, Pinctada fucata. Two isoenzymes (AcPase I and II) of acid phosphatase were separated and purified from viscera of pearl oyster, P. fucata to homogeneity by chromatography on DEAE-Sepharose Fast Flow, Sephadex G-200 superfine and ConA Sepharose 4B, and partial biochemical properties of AcPase I and II were studied. AcPase I and AcPase II had molecular weights of 208.8 and 64.3 kDa, respectively. AcPase I was a single polypeptide chain, while AcPase II was a dimeric enzyme composed of two equivalent subunits. AcPase I and II showed optimal pHs at 4.6 and 3.2 with p-nitrophenylphosphate as substrate. The optimal catalytic reaction temperature was 47 degrees C for AcPase I and 57 degrees C for AcPase II. Both enzyme forms were stable when incubated at 50 degrees C for 40 min. Tartrate and fluoride were the most effective inhibitors of the enzymes. Fe(3+), Zn(2+), Cu(2+) and Pb(2+) inhibited the activity of AcPase I and II to differing extents. AcPase I and II were apparently nonspecific and hydrolyzed various phosphoric esters. The different properties of AcPase I and II suggested that the two enzymes may play different roles in the pearl oyster. PMID- 16380282 TI - Effects of elevated ammonium on glycosylation gene expression in CHO cells. AB - The negative effects of ammonium on recombinant protein productivity and glycosylation have been well documented, but the interaction of ammonium on glycosylation genes has not been completely elucidated. In this study, the effects of elevated ammonium on 12 glycosylation related genes in Chinese hamster ovary cells were evaluated by quantitative real time reverse transcriptase polymerase chain reaction. Numerous cytosol and endoplasmic reticulum (ER) localized genes associated with early glycosylation steps were insensitive to the ammonium condition. The initial expression of uridine diphosphate (UDP)-galactose transporter was higher for the ammonium-treated culture, while the initial expressions of cytosine monophosphate (CMP)-sialic acid transporter, beta(1,4) galactosyltransferase, and UDP-glucose pyrophosphorylase were higher for the control culture. alpha(2,3)-sialyltransferase was observed to have lower expression level under the elevated ammonium condition compared to the control culture. This study indicates that galactosylation and sialylation inhibition is mainly due to decreased gene expression of galactosyltransferase, sialyltransferase, and CMP-sialic acid transporter and not due to sialidase. These unbalanced initial glycosylation and branching steps can explain the higher molecular heterogeneity under ammonium stress. Moreover, this study indicates that elevated ammonium has limited effects on the glycosylation genes associated with the ER and cytosol compared to the genes associated with the Golgi. PMID- 16380283 TI - [Chemotherapy in soft tissue sarcomas]. AB - Despite they represent an heterogeneous entity, the same protocols were applied to all subtypes of soft-tissue sarcomas until recently. Although doxorubicin and ifosfamide remain the cornerstone of therapy, their association yields enhanced response rates but has no obvious effect on survival. The benefit of adjuvant therapy is still matter of debate; however, it seems to improve relapse-free survival and might of particular interest for patients with high-grade tumours of the extremities. Yet, the major change occurring over the past few years is probably the development of subtype-specific regimens. Whether targeted therapies could provide additional benefit is a major concern but further studies are needed. PMID- 16380284 TI - [Place and technical aspects of external beam radiation therapy in the treatment of adult soft tissue sarcomas]. AB - In soft tissue sarcoma, surgical resection remains the cornerstone of therapy for localized disease. Quality of margins is very important to evaluate. In case of marginal or incomplete resection, a new enlarged surgical resection should always be discussed before administration of any adjuvant treatments. Many retrospective studies and 2 randomized studies (one of adjuvant brachytherapy and one of external beam radiotherapy) have shown that adjuvant radiotherapy after complete surgery reduces significantly the risk of local recurrence in extremity soft tissue sarcomas. Combination of surgery and pre- or postoperative radiotherapy has therefore become the standard treatment with a local recurrence rate .05). At discharge, patients who received slump stretching demonstrated significantly greater improvements in disability (9.7 points on the ODI, P < .001), pain (.93 points on the NPRS, P = .001), and centralization of symptoms (P < .01) than patients who did not. The results suggest that slump stretching is beneficial for improving short-term disability, pain, and centralization of symptoms. Future studies should examine whether these benefits are maintained at a longer-term follow-up. PMID- 16380287 TI - Predictors for the immediate responders to cervical manipulation in patients with neck pain. AB - Cervical manipulation has been considered an effective treatment for managing neck pain. However, clinical observation showed that cervical manipulation was not effective for every patient. Development of clinical prediction rules for identifying patients with neck pain who are likely to respond to cervical manipulation may improve clinical decision-making and the treatment success rate. The purpose of the study was to identify predictors for the immediate responders to cervical manipulation treatment in patients with neck pain. One hundred patients with neck pain (34 males and 66 females, mean age = 46 +/- 11 years) participated in the study. Patient's demographic data, symptom aggravating or easing factors, pain, and disability level were obtained through an initial assessment. A series of physical examinations were also administered. After receiving a single session of cervical manipulation, the patient was re-evaluated immediately to determine if a successful response to treatment was obtained. The successful response was determined by improvements seen in one of the three outcome variables that included reduction of pain intensity, significant perceived improvement, and high satisfaction level. From these judgment criteria, patients were classified into either responders or nonresponders to the cervical manipulation. Univariate analyses were used to assess if the treatment responders and nonresponders were different in their clinical presentations. The clinical factors that showed significant differences between two groups were then entered into a stepwise multiple logistic regression analysis to identify significant predictors and the prediction rule for treatment responders. Six predictors including "initial scores on Neck Disability Index < 11.50", "having bilateral involvement pattern", "not performing sedentary work > 5 h/day", "feeling better while moving the neck", "without feeling worse while extending the neck", and "the diagnosis of spondylosis without radiculopathy" were identified to significantly predict the immediate responders. The presence of four or more of these predictors increased the probability of success with manipulation to 89%. We concluded that using favourable predictors to identify treatment responders before administering cervical manipulations could significantly increase the probabilities of a successful treatment. This study finding may enhance the efficacy of clinical decision-making in using cervical manipulation intervention. PMID- 16380288 TI - Comparison of the effectiveness of a behavioural graded activity program and manual therapy in patients with sub-acute neck pain: design of a randomized clinical trial. AB - The objective is to present the design of a randomized clinical trial (RCT) on the effectiveness and cost effectiveness of a behavioural graded activity programme compared with manual therapy in patients with sub-acute neck pain. Sub acute is defined as pain existing for 4-12 weeks. The behavioural graded activity programme is a time-contingent increase in activities from baseline towards pre determined goals. Manual therapy consists mainly of specific spinal mobilization techniques and exercises. The primary outcomes are global perceived effect and functional status. Secondary outcomes are kinesiophobia, distress, coping, depression and somatization. The intensity and persistence of the pain and its interference with activities are also assessed. Direct and indirect costs are measured by means of cost diaries. Measurements take place at baseline and 6 and 12 weeks after randomization. To assess the long-term effect, measurements will also take place after 6 and 12 months. Finally some challenges are discussed concerning the use of a behavioural graded activity programme, manual therapy and outcomes. PMID- 16380289 TI - Primate orbitofrontal cortex and adaptive behaviour. AB - Orbitofrontal cortex contributes to behavioural adaptation in response to changes in the contingent relationship and incentive value of positive affective stimuli in the environment. This article integrates early descriptions of the effects of orbitofrontal ablation in monkeys, on object discrimination reversal and extinction, with contemporary theories of animal learning. Studies of incentive devaluation, conditioned reinforcement and changes in reward contingency are reviewed, highlighting the role of the orbitofrontal cortex in processing the affective and non-affective properties of rewarding stimuli, in reward expectation, and in goal selection. It is argued that future studies should focus on the interaction of the orbitofrontal cortex with peripheral arousal systems and the ascending monoamine systems in order to understand fully the role of the orbitofrontal cortex in behavioural adaptation. PMID- 16380290 TI - Challenges for control of taeniasis/cysticercosis in Indonesia. AB - Taeniasis/cysticercosis has been reported from several provinces of Indonesia: Papua (=former Irian Jaya), Bali, North Sumatra, East Nusa Tenggara, South East Sulawesi, Lampung, North Sulawesi, Jakarta, West Kalimantan, and East Java. The highest level of endemicity of taeniasis/cysticercosis has been found in Papua. Recent surveys in Jayawijaya District of Papua in 2000 and 2001 showed that 5 of 58 local people (8.6%) harbored the adult tapeworm, Taenia solium, whereas 44 of 96 people (45.8%), 50 of 71 pigs (70.4%), and 7 of 64 local dogs (10.9%) were seropositive for T. solium cysticercosis. Current surveys in Bali and Samosir District, North Sumatra during 2002-2005 revealed that Taenia saginata taeniasis has increased in incidence whereas T. solium cysticercosis is now rather rare compared to one-two decades ago in Bali. Taenia asiatica taeniasis is still common in Samosir District. Data from other provinces of Indonesia are very limited or unavailable. Control of these diseases is not a priority in the health or veterinary services, neither at central or local government levels. However, limited efforts toward control of the diseases have been implemented such as training of health personnel, community education on disease prevention, and provision of anthelminthics. A working group for control of the disease in Indonesia and an international collaboration have been established among Ministry of Health, Indonesia; University of Indonesia; and Asahikawa Medical College, Asahikawa, Japan since 1996. Future goals include implementation of active case finding (active surveillance) and treatment of tapeworm carriers, sustainable public health education, establishment of a system to check the quality of beef/pork and determine the distribution of infected animals and strengthening of laboratory capacity. Efforts to motivate provinces and districts should be implemented in developing the strategic plan to control of the disease. Given the considerable differences in cultures, religions, levels of education, socio economic levels, daily habits and behaviour etc., control programmes must be adapted to the local situations. PMID- 16380292 TI - IFNgammaR2 trafficking tunes IFNgamma-STAT1 signaling in T lymphocytes. AB - Ligand-dependent downregulation of the interferon gamma receptor signaling chain (IFNgammaR2) has always been seen as a key mechanism for shielding T lymphocytes from the antiproliferative effects of the IFNgamma-signal transducer and activator of transcription 1 (STAT1) pathway. Now, however, a ligand-independent mechanism of IFNgammaR2 internalization is emerging as a more general way of limiting IFNgamma-STAT1 signaling in T cells, with insulin-like growth factor-1 (IGF-1) and iron as the main players. Here, we review the array of immunomodulatory effects exerted by these two factors on different cell types involved in the immune response; these effects suggest that an inflammatory environment generates signals that favor IFNgammaR2 cell-surface accumulation and IFNgamma-induced apoptosis in T cells, whereas an anti-inflammatory environment promotes IFNgammaR2 internalization and induces T cell unresponsiveness to IFNgamma signaling. PMID- 16380293 TI - Dientamoebiasis: clinical importance and recent advances. AB - Dientamoeba fragilis, an unusual single-celled parasite that was described first in 1918, is found worldwide in the gastrointestinal tract of humans. D. fragilis has emerged from obscurity recently because it is now recognized as a common cause of chronic diarrhoea and is treatable with drugs. Recent molecular studies have described D. fragilis as having two genotypes. Diagnostic tests, based on conventional and real-time PCR, have been developed that will provide a rapid, sensitive and specific diagnosis of D. fragilis. These tests will also aid the elucidation of the host distribution and the life cycle of this pathogen. PMID- 16380294 TI - Immunopathogenic mechanisms in schistosomiasis: what can be learnt from human studies? AB - Studies in mice indicate that schistosome egg-induced granuloma formation and hepatic fibrosis depend markedly on cytokine regulation, with interleukin 10 having a central role. There is no clear consensus about the pattern of cytokine production and regulation that causes a minority of chronically exposed patients to develop severe hepatosplenic (HS) disease, which is characterized by periportal fibrosis and portal hypertension. HS disease and the progression of hepatic fibrosis are associated with the production of profibrotic type 2 cytokines in the early stages of infection with Schistosoma mansoni. However, other studies indicate that HS disease is characterized by a predominant T helper 1 profile. Until new tools and approaches are developed to study human disease in endemic areas, investigators must either speculate about indirect evidence from human studies or rely more heavily on findings generated from experimental models of the disease. PMID- 16380295 TI - Combined angiopoietin-1 and vascular endothelial growth factor gene transfer restores cavernous angiogenesis and erectile function in a rat model of hypercholesterolemia. AB - Hypercholesterolemia-related endothelial cell dysfunction and decreased endothelium-derived nitric oxide formation may account for impaired angiogenesis and subsequent erectile dysfunction. Angiopoietin-1 (Ang1) is a critical angiogenic factor for vascular maturation and enhances vascular endothelial growth factor (VEGF)-induced angiogenesis in a complementary manner. We hypothesized that combined adenovirus-delivered human Ang1 (ad-Ang1) and VEGF165 (ad-VEGF165) gene transfer might promote angiogenesis cooperatively in a rat model of hypercholesterolemic erectile dysfunction and result in a recovery of erectile function. Ad-Ang1 and ad-VEGF165 were injected either alone or in combination into the corpus cavernosum of the penis. Combined gene transfer of both ad-Ang1 and ad-VEGF165 significantly increased cavernous angiogenesis, eNOS phosphorylation, and cGMP expression compared with that in the groups treated with either therapy alone. Erectile function, as evaluated by electrical stimulation of the cavernous nerve 2 and 8 weeks after treatment, was completely restored in the combined treatment group, whereas intracavernous injection of either ad-Ang1 or ad-VEGF165 alone elicited partial improvement. The results indicate that combined application of angiogenic factors may enhance cavernous angiogenesis cooperatively by reinforcing the endothelium both structurally and functionally, which results in an additive effect on erectile function in hypercholesterolemic rats. PMID- 16380296 TI - Recurrent syncope in a case of metastatic malignant melanoma--an unusual presentation of an uncommon disease. AB - Cardiac tumours may be rare causes of syncope. Here we report a malignant melanoma that metastasized to the right atrium as an intracavitary mass that intermittently obstructed the tricuspid valve producing syncope episodes. PMID- 16380297 TI - Folic acid supplementation reduces the development of some blood cell abnormalities in children receiving carbamazepine. AB - BACKGROUND: Carbamazepine is a commonly used anticonvulsant agent; however, it has been linked with various blood cell abnormalities. This study evaluated the effect of low-dose folic acid supplementation on the prevention of carbamazepine induced hematological derangements in children. METHODS: This randomized clinical trial was conducted in children with epilepsy who received carbamazepine monotherapy. Group 1 received carbamazepine alone, and group 2 received carbamazepine plus folic acid. The two groups were age- and sex-matched. Each group comprised 41 children with epilepsy. Complete blood counts were obtained before starting medication (baseline) and then serially. The patients were followed for at least 1 year. RESULTS: In group 1, 31.4% of the patients developed leukopenia and 17.1% neutropenia, but in group 2, these figures were 14.6 and 9.8% (P = 0.067 and P = 0.331, respectively). At the end of the first year of follow up, white blood cell and polymorphonuclear cell counts were significantly higher in group 2 (P = 0.007 and P = 0.001, respectively). Hemoglobin concentration dropped in group 1, but rose slightly in group 2; these changes were significant. Platelet, lymphocyte, and monocyte counts and changes in serial blood tests did not differ significantly between the two groups. CONCLUSIONS: Folic acid is a safe drug that can reduce the development of some blood cell abnormalities linked to carbamazepine. It has a favorable effect on preventing the leukopenia and drop in hemoglobin observed in patients receiving carbamazepine, but its exact effect and the optimal dose required to enhance its benefits require further investigation. PMID- 16380298 TI - Oxidative stress and protective effects of polyphenols: comparative studies in human and rodent kidney. A review. AB - Reactive oxygen species (ROS) play a key role in the pathophysiological processes of a wide range of renal diseases. Thus, antioxidants are expected to decrease the vulnerability of the kidney to oxidative challenges. Polyphenols, particularly abundant in red wine, could act as ROS scavengers, iron chelators and enzyme modulators. In addition, chronic exposure to moderate amounts of ethanol results in increased activity of the renal antioxidant enzymes, further supporting a renoprotective effect of red wine based on its antioxidant properties. An enhancement of plasma antioxidant capacity following red wine consumption has been reported both in man and rodents, thereby providing a contributory factor to its renoprotective effect because the kidney is a highly perfused organ. Although phenol concentration of red wine does not influence the activity of antioxidant enzymes of the kidney, the concentration of these compounds is negatively correlated with tissue lipid peroxidation, assessed by thiobarbituric acid reactive substances, and positively correlated with the antioxidant capacity of plasma. Moreover, amelioration of myoglobinuric renal damage was found in rats following chronic exposure to flavonol-rich red wine. Also, pretreatment with resveratrol, or other red wine polyphenols, decreased kidney damage caused by ischaemia-reperfusion. The aim of the present review is to examine the pathophysiological basis of the renoprotective effect of red wine in man and rodents, based on functional, biochemical and ultrastructural evidence. PMID- 16380299 TI - Vitellogenin induction in the endangered goodeid fish Girardinichthys viviparus: vitellogenin characterization and estrogenic effects of polychlorinated biphenyls. AB - Vitellogenin (VTG) is a widely used biomarker in studies of endocrine disruption induced by xenobiotics such as polychlorinated biphenyls (PCBs). This study evaluates the estrogenic effects of these compounds on the black-fin goodeid Girardinichthys viviparus, an endangered fish species in Mexico with a reduced range of distribution due to pollution of its natural environment. Adult fish born in the laboratory were exposed to half the LC(0) of Inerteen commercial PCB mixture. VTG was determined through an inhibition enzyme-linked immunosorbent assay (ELISA) using a homologous-heterologous system. Male and female fish were killed after 1, 2, 4, 8 and 16 days of exposure. The distal third of each specimen was used for analysis. VTG was obtained from cultured hepatocytes and blood serum of males previously exposed to 17beta-estradiol. VTG molecular mass was 348 kDa. PCBs were found to elicit greater estrogenic effects on VTG induction in males than in females (p<0.05) and sex differences were noted. Time dependent VTG induction kinetics in males and a stationary phase in females were also observed. PMID- 16380300 TI - Homology assessment and molecular sequence alignment. AB - Hypotheses of homology are the basis of phylogenetic analysis. All character data are considered to be equivalent regardless of the source of those characters. Putative homology statements are designated based on observations of similarity. Pairwise sequence alignment using the Needleman-Wunsch algorithm is the basis for similarity maximization between molecular sequences. Multiple sequence alignment uses this algorithm in a topologically hierarchical framework. The resulting hypotheses of homology are tested in conjunction with character congruence through parsimony. This review introduces some underlying principles of phylogenetic analysis as they pertain homology testing and DNA sequence alignment. PMID- 16380301 TI - Determination of chlorhexidine (CHD) and nonylphenolethoxylates (NPEOn) using LC ESI-MS method and application to hemolyzed blood. AB - Rapid and reliable methods for identification of chlorhexidine (CHD) and nonylphenolethoxylates (NPEOn) in antiseptic and hemolyzed blood using electrospray ionization mass spectrometry (ESI-MS) were developed. Fragmental analysis provides accurate evidence for the presence of CHD in the samples. For the determination of CHD in hemolyzed blood, the method was also developed using LC-ESI-MS. Linearity of calibration curve was obtained over the concentration range of 0.1-11 microg/mL with residuals from -4.3 to 6.7%. We applied the methods to the case of suicidal injection of antiseptic and successfully detected CHD and NPEOn from hemolyzed blood. The CHD concentration was 352 microg/mL. PMID- 16380302 TI - Hemoglobin I from Lucina pectinata: a model for distal heme-ligand control. AB - Lucina pectinata hemoglobin I (HbI), which is a ferric sulfide-reactive hemeprotein, contains a distal pocket characterized by the presence of GlnE7 and PheB10. To elucidate the structural-functional properties of HbI, oxygen binding kinetics and FTIR studies with recombinant HbI (rHbI) and a set of mutants were conducted using CO and CN- as sensors of the hemeprotein environment. Three nuCO modes were observed for rHbI at 1936 cm(-1) (A3, closed conformer) 1950 cm(-1) (A1,2, closed conformer) and 1960 cm(-1) (A0, open conformer). These nuCO were affected by substitution of GlnE7 and PheB10 in the CO complexes. The contribution of GlnE7 is demonstrated when this residue is replaced with Asn, Val or His. For instance, decreasing the positive electrostatic environment with GlnE7Val, causes an increase of 65% in the population of A0 and the disappearance and 55% reduction of the population of the A1,2 and A3 respectively. The contribution of PheB10 to the stabilization of ligands is also observed in the Leu and Tyr mutants. The PheB10Leu mutation produced an 8% decrease in the population of the A3 conformer while that of the A1,2 configuration increased by 30%. This suggests that GlnE7 and PheB10 contribute to the A3 conformer stabilizing the CO in a closed configuration. With CN- as probe no substantial differences in the nuCN was observed upon substitution of GlnE7 by Val while a slight down shift in the nuCN from 2120 cm(-1) to 2117 cm(-1) was observed in the PheB10Leu mutant. This implies that in HbICN GlnE7 moves away from the binding site while PheB10 remains in the vicinity of the bound CN-. Here, a mechanism in which the flexibility of the distal protein matrix coupled with hemeporphyrin movement toward a different configuration is suggested as an important process in the H2S transport and delivery in hemoglobin I. PMID- 16380303 TI - The European Association of Consultation Liaison Psychiatry and Psychosomatics and its relationship to the journal. PMID- 16380304 TI - Psychosocial predictors of cardiac rehabilitation quality-of-life outcomes. AB - OBJECTIVE: This study investigated hostility, social support, coping, depression, and their contributions to concurrent and posttreatment quality of life (QoL) among a group of patients participating in a 6-week cardiac rehabilitation program. METHOD: Both direct and mediational relationships among psychosocial factors, QoL baseline, and QoL outcome were examined using structural equation modeling analysis, while age, education, and severity of illness (risk for future event) were controlled. RESULTS: The final model was well supported (chi(2)=64.88, df=56, P>.05; CFI=.99, RMSEA=.04). Results indicated that baseline QoL, hostility, and depressive symptom severity directly and independently predicted QoL outcome, while depression and hostility were also associated with baseline QoL. Hostility, social support, and maladaptive coping also contributed to baseline and follow-up QoL by their associations with depression. CONCLUSION: Psychosocial characteristics were interrelated, and they predicted postrehabilitation QoL outcome directly or indirectly through depression symptom severity. PMID- 16380305 TI - Depression following acute coronary syndromes: a comparison between the Cardiac Depression Scale and the Beck Depression Inventory II. AB - OBJECTIVE: This study compared the Cardiac Depression Scale (CDS) and the Beck Depression Inventory II (BDI-II). METHOD: Depression was assessed in 81 participants, 2 weeks post-ACS, using the BDI-II and the Composite International Diagnostic Interview. RESULTS: The CDS had a strong concurrent validity with the BDI-II (r=.69). Cross-validation of the BDI-II and the CDS with the structured interview demonstrated the ability of both measures to detect severe symptoms. More patients were classified as depressed using the CDS. The CDS also had a significantly higher correlation with a trait anxiety measure than the BDI-II did. CONCLUSION: The CDS is a more suitable scale for assessing the less severe depressive symptoms typically seen in a cardiac population. PMID- 16380306 TI - Depression, faith-based coping, and short-term postoperative global functioning in adult and older patients undergoing cardiac surgery. AB - OBJECTIVES: This prospective study examined how preoperative depression and faith based coping, assessed preoperatively and postoperatively, affected short-term postoperative global functioning (SPGF) following a major cardiac surgery. METHODS: We recruited 481 patients (male, 58%; mean age=62 years, range=35-89) 2 weeks before surgery for three sequential psychosocial interviews using standardized instruments. Of them, 426 completed the second interview, and 335 completed the postoperative follow-up. RESULTS: Multiple regression analyses showed that depression predicted poor SPGF, controlling for age, preoperative illness impact, and two noncardiac chronic conditions. Preoperative positive religious coping contributed to better SPGF, controlling for preoperative depression and other confounders. However, postoperatively assessed prayer coping was associated with poor SPGF. CONCLUSION: Research should distinguish the longitudinal protection of generally adaptive faith-based coping styles from the increased usage of such coping for immediate distress, mobilized by crisis. PMID- 16380307 TI - The prevalence and moderators of fatigue in people who have been successfully treated for cancer. AB - OBJECTIVE: The aims of this study were to estimate the prevalence of severe fatigue in disease-free breast cancer patients according to draft International Classification of Disease, Tenth Edition (ICD-10) criteria for cancer-related fatigue (CRF) and to obtain further information on the validity of these criteria. Furthermore, hypotheses derived from psychosocial theories of fatigue regarding the association of fatigue with activity level, psychological distress, and cognitive constructs were also tested. METHODS: Sixty-nine disease-free breast cancer patients were assessed at least 6 months posttreatment, using self report questionnaires and a structured interview. RESULTS: Nineteen percent of the sample met criteria for CRF. This subgroup differed significantly from the rest of the sample on multiple measures of fatigue and interference. Self reported activity level bore no relationship to fatigue. Fear of recurrence (FOR) contributed to fatigue indirectly, whilst psychological distress and beliefs about activity appeared to predict fatigue directly. CONCLUSION: The validity of the draft ICD-10 criteria for CRF was supported. Further research is required into the relationship between fatigue, emotional distress, and cognitive behavioural factors. PMID- 16380308 TI - Satisfaction with medical information in women with hypertensive disorders in pregnancy. AB - OBJECTIVE: Hypertensive disorders in pregnancy (HDP) are a major cause of maternal death and fetal prematurity. The purpose of this study was to examine HDP patients' reported satisfaction with the medical information provided by their health care providers. METHODS: A convenient sample of N=620 patients with a diagnosis of HDP within 24 months prior to the study period completed a self administered questionnaire dealing with the medical information received and their reported satisfaction. Chi-square, Fisher's Exact, and Student's t tests were used to determine the differences in sociodemographic data and potential confounders between HDP patients who were satisfied and dissatisfied with medical information. RESULTS: Of the women, 68.6% were not satisfied with the medical information. Women were particularly dissatisfied when they suffered from preeclampsia (PE), were informed of their actual situation, including eventual consequences only after delivery, and in case of fetal disorders resulting from HDP. CONCLUSION: Based on the background information reported on the importance of medical information to the patient, women developing HDP should be informed of their disease and potential sequelae as early and as differentiated as possible. Doctors need to be convinced of the role of the provider in counseling women with HDP. Information has to be repeated and health care providers are supposed to check whether the women understood the information received. PMID- 16380309 TI - Negatively oriented personality and perceived negative job characteristics as predictors of future psychological and physical symptoms: a meta-analytic structural modelling approach. AB - OBJECTIVES: There is evidence that perceptions of negative job characteristics (e.g., lack of control) and negatively oriented personality (e.g., negative affect) both predict symptom reporting. However, the relative contribution of these factors is rarely examined. This paper uses meta-analytic structural equation modelling (MA-SEM) to explore their relative contribution. METHODS: A meta-analysis of recent longitudinal studies was conducted. Eight studies covering 10 separate samples met the inclusion criteria (N=2104) and were used in the MA-SEM. Meta-correlations were used to construct structural equations models in which perceived negative job characteristics (PNJC) and negatively oriented personality (NOP) at baseline were used to predict concurrent and future symptom reporting, controlling for symptom reporting at baseline. RESULTS: The results indicated that a model based solely on NOP offered a more parsimonious account for baseline and future symptom reporting than did PNJC. CONCLUSIONS: The evidence indicates interventions should focus on both individuals and organizations, and not just organizational-level interventions. PMID- 16380310 TI - Spirituality predicts outcome independently of expectancy following flower essence self-treatment. AB - OBJECTIVE: The aim of this study was to determine whether absorption and spirituality predict the placebo response independently of expectancy. METHOD: This was an open study of self-treatment with self-selected Bach flower essences. Participants' expectancy of the effect of flower essences, attitudes to complementary medicine, holistic health beliefs, absorption, and spirituality were measured prior to treatment. One month after the start of treatment, participants responded to an e-mail enquiry about symptom change using a single seven-point change scale. RESULTS: One hundred sixteen participants (97 university undergraduates and 19 staff) completed all assessments. Spirituality and absorption together predicted additional variance compared with a cluster of expectancy measures comprising expectancy, attitude to complementary medicine, and holistic beliefs (increment in R(2)=.042, P=.032), and spirituality alone (but not absorption alone) predicted more additional variance than did the expectancy cluster (increment in R(2)=.043, P=.014). CONCLUSION: Our data are inconsistent with conventional explanations for the placebo effect. The mechanism underlying the placebo response is not fully understood. PMID- 16380311 TI - Effect of belief in "psychic healing" on self-reported pain in chronic pain sufferers. AB - OBJECTIVE: The "psychic healing" ability of a well-known Australian psychic was subjected to a televised experimental test. METHODS: Twenty volunteers suffering from chronic pain were recruited by newspaper advertisements. Half were randomly assigned to the treatment or control condition using a double-blind procedure. RESULTS: Comparison of pre- and posttreatment McGill Pain Questionnaire ratings indicated no effect of psychic healing. However, pretreatment questionnaire ratings of belief in psychic healing and related phenomena were significantly correlated with improvement in McGill Pain Questionnaire ratings irrespective of treatment condition. CONCLUSION: Results suggest that anecdotal reports of effective psychic healing and "faith healing" are attributable to the power of belief. PMID- 16380312 TI - Bullying at work, health outcomes, and physiological stress response. AB - The relationships among bullying or witnessing bullying at work, self-reported health symptoms, and physiological stress reactivity were analysed in a sample of 437 employees (294 women and 143 men). Physiological stress reactivity was measured as cortisol in the saliva. Of the respondents, 5% of the women (n=15) and 5% of the men (n=7) reported bullying, whereas 9% of the women (n=25) and 11% of the men (n=15) had witnessed bullying at work. The results indicated that the bullied respondents had lower social support from coworkers and supervisors, and they reported more symptoms of somatisation, depression, anxiety, and negative affectivity (NA) than did the nonbullied respondents. Witnesses reported more symptoms of anxiety and lower support from supervisor than did the nonbullied employees. Concentrations of cortisol in the saliva were lower at awakening in bullied respondents compared with nonbullied respondents. Previous studies have reported lower diurnal concentration of cortisol for people with posttraumatic stress disorder (PTSD) and chronic fatigue. To our knowledge, this is the first full study on the associations among being subjected to bullying, health outcomes, and physiological stress response. PMID- 16380313 TI - Learned helplessness in children and adolescents with juvenile rheumatic disease. AB - OBJECTIVE: To examine a learned helplessness conceptualization of psychological sequela in children and adolescents with juvenile rheumatic diseases (JRD) via an experimental procedure utilizing behavior-outcome contingent and noncontingent feedback. METHODS: Thirty-eight children and adolescents with JRD completed measures of transient affect, self-efficacy for functional ability, and causal attributions prior to and immediately following a computerized learned helplessness induction procedure. RESULTS: Children across contingent and noncontingent feedback conditions experienced decreased positive affect with a slightly more pronounced decline in the noncontingent condition. Noncontingent feedback resulted in poorer internalization of success for problem solving, while contingent feedback resulted in greater internalization of success for problem solving. Additionally, posttreatment control self-efficacy was significantly greater for children in the contingent condition that initially endorsed higher levels of internal task attributions. CONCLUSIONS: Children with JRD who experience behavior-outcome contingency in their environment may develop increased perceptions of control over functional ability. Furthermore, environmental noncontingency may result in poorer internalization of success, whereas contingent reinforcement may enhance cognitive appraisal mechanisms (i.e., causal attributions) associated with favorable disease outcome. Despite a modest decline in positive affect for children in the noncontingent condition, mood dysfunction is not entirely explicable within the context of noncontingent reinforcement. PMID- 16380314 TI - Preliminary evidence of parasympathetic influence on basal heart rate in posttraumatic stress disorder. AB - OBJECTIVE: Evidence of elevated basal heart rate (HR) in posttraumatic stress disorder (PTSD) has been interpreted in terms of elevated sympathetic cardiac activity, as have possible increased cardiovascular disease risks and outcomes associated with elevated HR. Yet it is well-established that the parasympathetic branch of the autonomic nervous system not only influences HR independently of the sympathetic branch, but makes a greater contribution to HR, including resting HR. Additionally, abnormally low tonic parasympathetic activity on the heart has been implicated in cardiovascular disease and hypertension. This study addressed a potential parasympathetic contribution to elevated basal HR in PTSD. METHODS: We used a descriptive and subgroup differences approach to investigate relationships between parasympathetic activity and basal HR in 59 adults (50 females) with PTSD, all of whom were participants in a treatment outcome study and assessed prior to exposure to trauma-related script-driven imagery. Consistent with the well-known relationship between parasympathetic activity and HR, we hypothesized that basal respiratory sinus arrhythmia (RSA), a measure of parasympathetic cardiac activity, would be negatively correlated with basal HR. More important, we predicted that pathologically elevated HRs previously associated with PTSD would only characterize a low-RSA subgroup. Potential confounds of age, respiration rate, and aerobic fitness were addressed. RESULTS: As predicted, mean HR was 80.5 bpm in the low-RSA tercile group, similar to mean HRs of PTSD groups in prior research and significantly higher than 69 and 65 bpm in the middle- and high-RSA groups, respectively, which are typical of non-PTSD controls. CONCLUSION: These findings suggest that a substantial proportion of those with PTSD may not have elevated basal HRs. Furthermore, among those who do exhibit elevated HR, there may be a parasympathetic contribution that is independent of any sympathetic one. Only measuring both branches at once, ideally with autonomic blockades, can definitively address these issues. PMID- 16380315 TI - Gender difference in association between polymorphism of serotonin transporter gene regulatory region and anxiety. AB - OBJECTIVE: The objective of this study was to verify the hypothesis that variation of the serotonin transporter gene promoter region (5-HTTLPR) is associated with sensitivity to stress. METHODS: Genotyping of 5-HTTLPR and evaluation of emotional states were performed on 194 participants. Participants' emotional states were evaluated using the Perceived-Stress Scale (PSS), the State Trait Anxiety Inventory (STAI), and the Self-rating Depression Scale (SDS). RESULTS: There was significant GenderxGenotype interaction in STAI (state, P<.05; trait, P<.05). Females with the l/s genotype showed higher anxiety than those with the s/s genotype in both state and trait anxiety. Oppositely, males with the s/s genotype showed higher anxiety than those with the l/s genotype. CONCLUSION: On all emotional scales, females with the l/s genotype showed high scores, contrary to males with the same genotype. Therefore, our results suggest that 5 HTTLPR l allele may be one pathway that activates negative emotion in females but acts contrary in males. PMID- 16380316 TI - Relationship between plasma concentrations of cytokines, ratio of CD4 and CD8, lymphocyte proliferative responses, and depressive and anxiety state in bulimia nervosa. AB - OBJECTIVE: Immunological dysfunction in participants with bulimia nervosa (BN) might be due to not only chaotic eating behavior but also psychological state, such as depression and anxiety, because studies have found a close relationship between depression, anxiety, and immunological dysfunction. METHOD: Participants included 20 females with BN and 14 control females. In eight BN participants, immune function was measured twice, before and after inpatient treatment. Cytokines [interleukin (IL)-1 receptor antagonist, IL-2 soluble receptor alpha, IL-6 soluble receptor (sIL-6r), and tumor necrosis factor soluble receptor II (sTNF-rII)], lymphocyte subsets (CD4 and CD8), and lymphocyte proliferative responses were measured. RESULTS: There were no differences in plasma cytokine levels or lymphocyte subsets between the BN participants and the controls. Conversely, lymphocyte proliferative responses were significantly lower in BN participants than in the controls. Lymphocyte proliferative responses negatively correlated with anxiety trait and improved with an improvement of bulimic state. CONCLUSION: The comorbid nonspecific anxiety trait significantly contributes to suppressing lymphocyte proliferative responses in BN participants. PMID- 16380317 TI - Bulimia nervosa and abnormal cardiac repolarization. AB - OBJECTIVE: Eating disorders (ED) are thought to be risk factors for sudden death, and arrhythmias are one of the major causes of sudden death in ED patients. Late potentials (LPs) are a predictor of arrhythmias and can be measured using signal averaged electrocardiography (SAECG). We examined arryhthmogenicity by LPs in ED patients. METHODS: We performed SAECG on 48 female ED patients [21 with anorexia nervosa (AN) and 27 with bulimia nervosa (BN)] and on 20 healthy women. An LP was judged positive if two or more of the following criteria were fulfilled: QRS duration >120 ms, root-mean-square voltage <20 microV, and a high-frequency, low amplitude duration >38 ms. We compared the occurrence of LPs among subgroups. RESULTS: Compared with the controls, BN patients with a history of AN had significantly more SAECG abnormalities. CONCLUSIONS: BN patients with a history of AN may be prone to ventricular arrhythmias. PMID- 16380318 TI - Coping with emotions and abuse history in women with chronic pelvic pain. AB - OBJECTIVE: The purpose of this study was to investigate whether past abuse and the tendency to repress or suppress unwanted thoughts and emotions contribute to the experience of pain in patients with chronic pelvic pain (CPP). METHODS: A group of CPP patients without endometriosis and a group with endometriosis were compared with a pain-free control group. Participants completed measures of pain, emotional repression, suppression of unwanted thoughts and emotions, and past abuse history. RESULTS: Both CPP groups were more likely to be emotional suppressors when compared with the control group and reported significantly higher levels of thought suppression and abuse. Endometriosis patients were also more likely to be repressors of emotions when compared with controls. Suppression but not repression was related to higher levels of abuse and pain. CONCLUSION: Suppression of unwanted thoughts and emotions and past abuse distinguishes CPP patients from healthy controls. Assisting patients to express distressing emotions may impact on pain levels. PMID- 16380320 TI - Pathogenesis of acute experimental liver amebiasis. AB - Classical descriptions of the pathology of amebiasis portray the parasite as the cause of tissue damage and destruction, and in recent years a number of amebic molecules have been identified as virulence factors. In this review we describe a series of experiments that suggest a more complex host-parasite relation, at least during the early stages of acute experimental amebic liver abscess in hamsters. The problems of extrapolating experiments in vitro to explain observations in vivo are discussed. The role of amebic cysteine proteases is examined and evidence presented to suggest that they are primarily related not to tissue damage but to amebic survival, which is required for the progression of the lesion. Inflammation is shown to be not only the major cause of tissue damage but also an absolute requirement for amebic survival in the liver, whereas complement and ischemia are not involved in the disappearance of the parasite in the absence of inflammation. PMID- 16380321 TI - Experimental amebiasis: a selected review of some in vivo models. AB - The use of in vivo animal models in amebiasis has contributed significantly to the knowledge of this common human parasitic disease. Although there is no animal model that mimics the whole cycle of the human disease, the use of different susceptible and resistant laboratory animals and the availability for many years of techniques for the axenic culture of trophozoites of Entamoeba histolytica have allowed a better understanding of the parasite and the host-parasite relationship. The recent introduction of frontier methodologies in biology has increased our comprehension of this parasite. New information on the cellular and molecular biology and genetics of this organism has been extensively reported, and much of this has clearly required the more frequent use of animal models to verify specific facts. Based on experimental animals characterized previously, the introduction of new animal models with genetic or surgical modifications, especially in mice, has allowed a more adequate analysis of the mechanisms of pathogenesis. Multiple factors have been considered in the promotion of the invasiveness and virulence of E. histolytica. Additionally, the immunological and physiological responses of the host, depending on the environmental conditions, lead to the establishment or the rejection of the parasite. The role of inflammatory reaction to amebic infection constitutes one of the controversies that has been studied by several authors. In susceptible animals (hamsters and gerbils), inflammatory cell damage seems to be related to target cell lysis, while in resistant animals (mice), inflammatory cells appear to protect the host by lysing the parasite. Presently, the involvement of various substances in the development of lesions including lectins, proteases, amebapores, promoters of apoptosis, cytokines, nitric oxide, etc., is being examined using different in vivo models. PMID- 16380322 TI - Calcium-binding proteins of Entamoeba histolytica. AB - Calcium plays an essential role in many fundamental processes in almost all eukaryotic cells including protozoan parasite Entamoeba histolytica. Many of the calcium-mediated processes are carried out through the help of calcium-binding proteins (CaBPs). A few of these E. histolytica CaBPs have been described before. These proteins are unique to this organism and are thought to be essential. Availability of genome sequence has opened up the possibility of studying CaBPs at the whole genome level. In this preliminary report, we describe the complement of CaBPs present in E. histolytica. A large fraction of these genes are expressed in the trophozoites and are likely to be functional. The results suggest a number of pathways that are involved in calcium signaling and may be unique for this organism. PMID- 16380323 TI - Epigenetic silencing of gene expression in Entamoeba histolytica. AB - Transcriptional silencing of an amebapore (ap-a) gene occurred in Entamoeba histolytica following the transfection of plasmids containing a DNA segment (473 bp) homologous to the 5' upstream region of the gene. This segment contains the promoter region of the ap-a gene, a T-rich stretch, followed by a truncated SINE1 (short interspersed element) that is transcribed from the opposite strand. The downstream silencing of the ap-a gene did not occur with plasmids containing the entire SINE1 sequence or lacking the entire SINE1 sequences including the T-rich stretch. Such plasmids promoted the overexpression of the ap-a gene. The transcription of the SINE element required both the T-rich stretch as well as sequences from the 5' end of SINE. RNA extracts from gene-silenced cultures showed small amounts of short (approximately 140 nt), single-stranded molecules with homology to SINE1 transcripts but no siRNA. Chromatin immunoprecipitation (ChIP) analysis of silenced G3 trophozoites with an antibody against methylated K4 of histone H3 revealed a demethylation of K4 at the domain of the ap-a gene indicating transcriptional inactivation. These results suggest the involvement of the SINE1 element in triggering the gene silencing and the role of histone modification in its epigenetic maintenance. The avirulent phenotype of the silenced trophozoites was demonstrated in various assays and the results suggest they may have a potential use for vaccination. PMID- 16380324 TI - The cytoskeleton of Entamoeba histolytica: structure, function, and regulation by signaling pathways. AB - Pathogenesis in the parasite Entamoeba histolytica has been related to motility of the trophozoites. Motility is an important feature in amebas as they perform multiple motile functions during invasion of host tissues. As motility depends on the organization and regulation of the cytoskeleton elements, in particular of the actin cytoskeleton, the study of the molecular components of the machinery responsible for movement has been a key aspect to study in this parasite. Although many of the components have high homology in amino acid sequence and function to those characterized in higher eukaryotic cells, there are important differences to suggest that parasitic organisms may have developed adaptative differences that could be useful as targets to stop invasion. The purpose of this review is to evaluate current knowledge about the cytoskeleton of E. histolytica and the ways in which the parasite controls motility. PMID- 16380325 TI - New insights into molecular mechanisms of phagocytosis in Entamoeba histolytica by proteomic analysis. AB - The protozoan parasite Entamoeba histolytica ingests microorganisms and mammalian cells. Phagocytosis is essential for cell growth and is implicated in pathogenesis of E. histolytica. Phagocytosis consists of a number of steps including recognition of and binding to ligands on the target cells via a galactose/N-acetylgalactosamine-specific lectin, activation of a signaling pathway leading to cytoskeletal reorganization, and vesicle trafficking, all of which play distinct but coordinated roles in phagocytosis. Recent studies of proteomic analysis of purified phagosomes or affinity-purified Gal/GalNAc-binding proteins using reversed phase capillary liquid chromatography and ion trap tandem mass spectrometry enabled high throughput identification of proteins involved in phagosome biogenesis. These studies provided a list of proteins involved in the pathway and also shed light on the dynamic process of phagosome maturation. These approaches should provide significant insights into molecular mechanisms of phagosome biogenesis and help to elucidate the pathogenesis of this important parasite. PMID- 16380326 TI - Host tissue invasion by Entamoeba histolytica is powered by motility and phagocytosis. AB - During amebiasis, E. histolytica motility is a key factor to achieve its progression across tissues. The pathogenicity of E. histolytica includes its capacity to phagocyte human cells. Motility requires polarization of E. histolytica that involves protrusion of a pseudopod containing actin and associated proteins [myosin IB, ABP-120 and a p21-activated kinase (PAK)] and whole-cell propulsion after contraction of the rear of the cell, where myosin II and F-actin are concentrated. An interesting characteristic of this parasite is the presence of two unique myosins (myosin II and unconventional myosin IB), in contrast to several actin genes. Little is known about the regulation of the actin-myosin cytoskeleton dynamics of E. histolytica, and a better understanding of signaling pathways that stimulate and coordinate regulators protein and cytoskeleton elements will provide new insight into the cell biology of the parasite and in amebiasis. Here we summarize the pleiotropic functions described for myosin II and PAK in E. histolytica. We propose that survival and pathogenicity of E. histolytica require an active actin-myosin cytoskeleton to cap surface receptors, to adhere to host components, to migrate through tissues, to phagocyte human cells and to form liver abscesses. PMID- 16380327 TI - Methods for the investigation of diversity in Entamoeba histolytica. AB - The ability to distinguish variants of a species has many potential applications. In Entamoeba histolytica the first method to detect variation was based on isoenzyme analysis. However, this approach has been superseded by DNA-based analysis. In this review I discuss the basis of the variation detected in E. histolytica by the various molecular methods that have been published to date. Information on diversity in other species is mentioned where such information exists. PMID- 16380328 TI - Epidemiology of amebiasis in Mexico: a molecular approach. AB - The differentiation of the two Entamoeba species E. histolytica and E. dispar through molecular characterization in the last decade of the twentieth century led to the need to re-evaluate the epidemiology of amebiasis in terms of prevalence and morbidity of the infection in the world's population, particularly in those geographic regions with high endemic rates. PMID- 16380329 TI - Prevalence and incidence of Entamoeba histolytica infection in South Africa and Egypt. AB - There are little data on the true prevalence and incidence of Entamoeba histolytica infection in Africa. This is due to the inability, historically, to differentiate Entamoeba histolytica from the more common, but non-pathogenic, Entamoeba dispar. In addition, newer studies have demonstrated that the previous gold standard, culture with zymodeme analysis, is insensitive in detecting the presence of infection, especially when compared to PCR. Recent published articles as well as data from the authors' previous work are reviewed and summarized to elucidate what is known about prevalence and incidence of Entamoeba histolytica in Africa. The majority of data on asymptomatic infection are published from South Africa, Egypt and Cote d'Ivoire. Egypt has high rates of asymptomatic infection detected in the stool (>21%), whereas South Africa and Cote d'Ivoire rates range between 0 and 2%. Seroprevalence estimates the rate of recent infection, because anti-amebic antibodies generally persist for <5 years. Seropositivity rates (IgG, IgA) range from approximately 10 to 20%, indicating recent infection in this proportion of the population. Entamoeba histolytica infects a significant proportion of many populations of Africa; however, little data are currently available to indicate true prevalence and incidence. Further studies are needed to determine the burden of infection and disease in Africa. PMID- 16380330 TI - Epidemiology and treatment of amebiasis in Hue, Vietnam. AB - Hue, a city of about 300,000 inhabitants in Central Vietnam, is known as a "hot spot" for diseases caused by Entamoeba histolytica and in particular for amebic liver abscess (ALA), thus representing an excellent site for studies on the epidemiology and treatment of E. histolytica infections. Accordingly, during the last few years, various studies on this topic have been performed in Hue in close collaboration among the Hue Medical College, the Hue Central Hospital and the Bernhard Nocht Institute for Tropical Medicine, Hamburg, Germany. These studies comprised (i) comprehensive analyses of a considerably large number of ALA cases, (ii) parasitological and seroepidemiological surveys on the prevalence and outcome of E. histolytica infections as well as (iii) comparative randomized treatment trials in order to optimize the therapeutic regimen for ALA and for asymptomatic E. histolytica infection. PMID- 16380331 TI - Diagnosis of amebiasis in Bangladesh. AB - Diagnosis of amebiasis by microscopic identification of the parasite in stool and liver abscess pus is insensitive and unable to distinguish the invasive parasite E. histolytica from the commensal parasites such as E. dispar and E. moshkovskii. New approaches to the detection of E. histolytica are based on detection of E. histolytica-specific antigen and DNA in stool and other clinical samples. Several molecular diagnostic tests for diagnosis of amebiasis have been developed and used to diagnose E. histolytica in Bangladesh. We have compared the TechLab E. histolytica-specific antigen detection test with PCR assays and with isoenzyme analysis of cultured amebas. The PCR assays are based on amplification of the multi-copy small subunit ribosomal RNA gene of E. histolytica and E. dispar. PCR assays and antigen detection test had comparable sensitivities when performed directly on fresh stool specimens. The correlation of antigen detection with PCR assays for identification of E. histolytica was excellent. TechLab's E. histolytica- specific antigen detection test was both rapid and simple to perform, making it appropriate for use in the developing world, where amebiasis is most prevalent. PMID- 16380332 TI - Diversity of clinical isolates of Entamoeba histolytica in Japan. AB - In Japan, amebiasis is domestically transmitted by two major populations: male homosexuals and mentally handicapped persons, which is remarkably different from most other developed countries where Entamoeba dispar infection is predominantly observed. Here we briefly summarize epidemiology of amebiasis in Japan. We also review our current understanding of the diversity of Entamoeba histolytica clinical isolates in Japan, based on polymorphic genetic markers, clinical representations, and in vivo virulence, using an animal model. PMID- 16380333 TI - Recent progress in vaccines for amebiasis. AB - The persistence of amebiasis as a global health problem, despite the availability of effective treatment, has led to the search for vaccines to prevent this deadly disease. Recent clinical studies suggest that mucosal immunity could provide some protection against recurrent intestinal infection with E. histolytica, but there is contradictory evidence about protective immunity after amebic liver abscess. Progress in vaccine development has been facilitated by new animal models that allow better testing of potential vaccine candidates and by the application of recombinant technology to vaccine design. Oral vaccines utilizing amebic antigens either co-administered with some form of cholera toxin or expressed in attenuated strains of Salmonella or Vibrio cholera have been developed and tested in animals for mucosal immunogenicity. Although there has been significant progress on a number of fronts, there are unanswered questions regarding the effectiveness of immune responses in preventing disease in man and, as yet, no testing of any of these vaccines in humans has been performed. PMID- 16380334 TI - Adherence-blocking vaccine for amebiasis. AB - The Gal/GalNAc lectin is a candidate vaccine antigen for an amebiasis vaccine due to its mediation of parasite adherence to the human intestine, because partial immunity in humans is associated with a mucosal IgA response against it, and because it is effective as a vaccine against amebic colitis in the murine model. The LecA domain of the Gal/GalNAc lectin contains neutralizing antibody epitopes. LecA contains the active site of the lectin (the carbohydrate recognition domain or "CRD") and has been an effective vaccine antigen in animal models of amebic colitis and liver abscess. Research needs include production of the LecA domain of the Gal/GalNAc lectin by a process that can be transferred to cGMP and optimization for immunogenicity, using adjuvants such as alum, MF59 or QS-21 adjuvants. Accomplishing this will enable testing of the ability of LecA immunizations to protect from amebic colitis in humans. PMID- 16380335 TI - The current status of an amebiasis vaccine. AB - Efficient control of infectious diseases requires the development and application of suitable vaccines. Development of vaccines against amebiasis is still in its infancy. However, in recent years progress has been made in the identification of possible vaccine candidates, the route of application and the understanding of the immune response that is required for protection against amebiasis. PMID- 16380336 TI - Antibiotic principles from a Streptomyces species and their sub-acute toxicity studies on hepatic, renal and haemopoietic system of rats. AB - Two promising antibiotics, JF-A and JF-B were isolated from the chloroform extract of a Banglaeshi Streptomyces strain. The mean zones of inhibition produced by the chloroform extract (400 microg/disc), JF-A (200 microg/disc) and JF-B (200 microg/disc) against 19 pathogenic bacteria were found to be 9-50, 12 38 and 10-41 mm while those produced by a standard antibiotic, kanamycin were 11 40 mm at 30 microg/disc. MICs of JF-A and JF-B were determined to be 64 microg/ml against Bacillus subtilis and 64 and 128 microg/ml against Shigella sonnei, respectively. The extract and the antibiotics were also tested for cytotoxicity against Artemia salina nauplii and LC50 values of 23.26, 18.05 and 32.27 microg/ml were obtained. 90% mortality of shrimp nauplii was observed at 69.18, 50.12 and 110.91 microg/ml, respectively. In a potato disc bioassay, the chloroform extract at 25 microg/disc demonstrated 37.39% inhibition of crown gall tumour induced by G -ve Agrobacterium tumefaciens B6 and the result was statistically significant (P < 0.05). The sub-acute toxicity studies on the JF-A and JF-B reflected innocuous nature of these antibiotics on hepatic, renal and haemopoietic system of rats at 1 mg/kg b.w. on daily administration for 21 consecutive days. This is also confirmed by detailed histopathological studies. No mortality was observed in experimental animals. PMID- 16380337 TI - Determination and quantification of cetirizine HCl in dosage formulations by RP HPLC. AB - A simple, sensitive, reliable and rapid HPLC method for the determination of cetirizine hydrochloride using hyoscine butyl bromide as an internal standard has been developed. The chromatographic system consisted of Shimadzu LC-10 AT VP pump, SPD-10 AV VP with UV/visible detector and a CBM-102 Bus Module integrator. Separation was achieved on the U Bondapak 125 A C18 10 microm column at room temperature. The samples were introduced through an injector valve with a 10 microl sample loop. Acetonitrile-water (1:1 v/v) was used as mobile phase, with flow rate 2 ml/minute. pH was adjusted to 2.9 with phosphoric acid. UV detection was performed at 205 nm. The results obtained showed a good agreement with the declared content. Recovery values for cetirizine hydrochloride were 99.19 100.82%. The proposed method is reliable rapid, precise, selective and may be used for the quantitative analysis of cetirizine HCl, in presence of hyoscine butyl bromide as internal standard. The method was valid was for the determination in raw materials, bulk drug and formulations. The limit of quantification was 5-30 nano grams, while the limit of detection was 0.4 nano grams. PMID- 16380338 TI - Antibiotic resistance pattern among gram-negative bacteria isolated from raw milk in Multan. AB - The use of raw milk in Pakistan has led to the antibiotics resistance. This investigation has showed incidence and antibiotic resistance among Coliform and Klebsiella to nalidixic acid, urixin and little to chloramphenicol. Enterobacter was sensitive to nalidixic acid, tetracycline and showed little resistance to chloramphenicol. E. coli were very sensitive to nalidixic acid, tetracycline, chloramphenicol, urixin, amoxil and with no resistance to ceftriaxone. PMID- 16380339 TI - Anti-inflammatory activity of Coldenia procumbens Linn. AB - Anti-inflammatory activity of the ethanolic extract of the aerial parts of Coldenia procumbens Linn. was studied in Wister rats using the carrageenan induced left hind paw edema, carrageenan induced pleurisy and cotton pellet induced granuloma model. The ethanolic extract (150 mg/kg, p.o.) produced the inhibition of carrageenan induced rat paw edema. It also showed an inhibitory effect on leukocyte migration and a reduction on the pleural exudates as well as reduction on the granuloma weight in the cotton pellet granuloma method. The results indicated that the ethanolic extract produced significant (P<0.001) anti inflammatory activity when compared with the standard and untreated control. PMID- 16380340 TI - Kinetics of copper(II) catalyzed oxidation of iodide by iron(III) orthophenanthroline complex in aqueous solution. AB - Orthophenanthroline (OPT) forms stable complex with Iron(II) at pH 3.5. In presence of Ce(IV) it forms a metal-ligand complex in which Fe is in the form of Fe(+++) for short period when mixed in mole ratio of 1:3 (Iron: OPT). This complex is stable for not more than 30 minutes. At the end, this complex reduces itself back to iron(II)orthophenanthroline [Fe(OPT)3](+2) complex. In present work, this property of iron(III) complex was used to oxidize I(-1) to I2. Analysis of [Fe(OPT)3](+2) was carried out at 510 nm to monitor the reduction of [Fe(OPT)3](+3) complex. Copper was used as a catalyst during the described reaction to convert back [Fe(OPT)3](+2). The reaction increased with increase in copper concentration. The conversion of [Fe(OPT)3](+3) to [Fe(OPT)3](+2) followed first order kinetics. PMID- 16380341 TI - The influence of different plasticizers on some physical and mechanical properties of hydroxypropyl methylcellulose free films. AB - Coating has been widely used in pharmaceutical manufacture either as non functional or a functional entity. The objectives of the present study were to investigate the effect of plasticizers such as PEG400, PEG1000 and triacetin on mechanical properties, glass transition temperature and water vapor transmission of free films prepared from HPMC and/or HPMC:PVA blends, to develop suitable coating system for tablets, and to determine the release profiles of the coated tablets. The tensile strength of plasticized HPMC films was generally lower than that of control HPMC film and could be attributed to increased crystallinity and segmental chain mobility of HPMC. This effect increased as the concentration of plasticizer increased. Generally the addition of both grades of polyethylene glycol (PEG400 & PEG1000) increased the moisture permeability of HPMC films but the films containing triacetin provided a more rigid barrier to moisture compared to unplasticized HPMC films. The dissolution profiles of paracetamol tablets coated with 7% w/v HPMC coating-solutions containing PEG400, PEG1000 and triacetin, and those containing PEG400 & PVA together showed that HPMC had weak water resistance. The presence of PEG400 and 1000 in HPMC films further weakened its resistance to solubility while the presence of triacetin caused a little increase in HPMC water resistance. From the results it was concluded that HPMC at 7%w/w concentration was suitable for film-coating intended for non-functional coating. Presence of the PEG 400, PEG1000 and triacetin as well as the presence of PVA and PEG400 together improved the coating properties of HPMC films and made it more suitable as a non-functional coating material. PMID- 16380342 TI - Antibacterial activity of 1-methyl-7-methoxy-beta-carboline and its phenacyl and coumarine analogues. AB - Antibacterial activity of 1-methyl-7-methoxy-beta-carboline (harmaline) and its phenacyl and coumarine analogues 1-(3-nitro-phenyl)-(2-(7-methoxy-1-methyl 1,3,4,9-tetrahydro-beta-carbolin-2-yl)-ethanone (II), 1-(3,4-dihydroxy-phenyl)-2 (7-methoxy-1-methyl-1,3,4,9-tetrahydro-beta-carbolin-2-yl)-ethanone (III) 7 (methoxy-beta-carboline),15-24,de-hydro(19,20-dimethoxy)coumarine (IV), 7 (methoxy-beta-carboline)15-24,dehydro(20-methoxy)coumarine (V) were studied by disc diffusion method. All compounds were tested against three gram positive and four gram-negative bacteria. Parent compound showed good activity. All compounds revealed better results against gram positive as compared to gram-negative bacteria. 1-(3,4-Dihydroxy-phenyl)-2-(7-methoxy-1-methyl-1,3,4,9-tetrahydro-beta carbolin-2-yl)-ethanone (III) was found most potent compound showing broad spectrum activity when compared with all synthesized analogues. Coumarine analogues showed more or less same activity indicating that number and position of methoxy groups are not important regarding antimicrobial activity. PMID- 16380343 TI - The effects of acetylsalicylic acid on the pituitary prolactin of the lizard, Uromastix hardwickii. AB - This study deals with the intravenous administration of 7 mg acetylsalicylic acid (ASA) solution to Uromastix hardwickii for 4 days. It enhances the activity of anterior pituitary lactotrophs, when 0.1 ml of pituitary homogenate of ASA treated was injected hypodermically to crop-sac showed a greater diametric response and increased activity with milk like secretion than that of the injections of 0.1 ml homogenate of control pituitary. The present study indicated that ASA induces hyperprolactinemia. PMID- 16380344 TI - Inhibition of proteinase 3 (PR3) by suramin and fetal calf serum (FCS): effect of PR3 and suramin on Chinese hamster ovary cells (CHO-cells). AB - BACKGROUND: Proteinase 3 (PR3) is a lysosomal protease that is stored in azurophilic granules neutrophilic granulocytes and monocytes. A number of inhibitors for this proteinase are reported. Comprehensive studies on the inhibitory effect of suramin and heat treated fetal calf serum (deltaFCS) on PR3 have not been reported. It has been reported that PR3 is able to destroy the cytoskeletal integral proteins, but we have not find any reports which showed the effect of this protease on Chinese hamster ovary cells (CHO-cells) in culture medium. Suramin has proven to be useful as an antitumor drug, but there was not any report on the effect of suramin on CHO-cells. METHODS: The effects of various concentrations of deltaFCS (from 0.5% up to 10%) and suramin (from 0.8 microM up to 100 microM) on PR3 and different concentrations of suramin (from 0.8 microM up to 1000 microM) on CHO-cells were investigated. Data analysis were performed by, Kolmogorov-Smirnov test, ANOVA test and Tukey HSD post tests. RESULTS: Results showed that deltaFCS and suramin have an inhibitory effect on PR3 and these effects increased with increasing the concentration significantly (p < 0.01). PR3 with the concentration of 2.2 Unit/ml has no effect on CHO-cells. Although suramin with the concentration of less than 125 microM cell growth retarded for only a few hours, but with the concentration of 125 to 250 microM inhibit the cell growth for a week, and after that cells gain normal growth gradually. Suramin with concentration of more than 500 microM inhibited the cell growth completely. CONCLUSIONS: Although suramin reversibly inhibit the PR3 activity but in concentration of less than 250 microM it had no long-term effect on CHO-cells. Therefore it can be used in the investigation of proteases. There were unknown components in deltaFCS, which cause the inhibition of PR3 activity. This finding is very important in PR3 production in culture medium. However CHO-cells are resistant to PR3 and suramin in low concentration. PMID- 16380345 TI - Brain tryptophan metabolism remained unaltered in restraint stressed rats following fluoxetine administration. AB - Effects of two hours immobilization stress on tryptophan (Tp) metabolism in rats was studied following fluoxetine-HCl (20 mg/kg) administration. After 2 hr immobilization stress there was no effect on peripheral liver Tp metabolism but there was marked increase in brain Tp metabolism leading to increases in 5-HT (5 Hydroxy tryptamine) synthesis and turn over. Rats subjected to 2 hrs immobilization stress immediately after fluoxetine-HCl (20 mg/kg) administrations show significant increases in holo and total Tp pyrrolase enzyme activities leading to decreased serum total Tp concentrations. Brain Tp and 5-HT remain unchanged but 5-hydroxy indole acetic acid (5-HIAA) concentration was significantly increased when compared to fluoxetine injected non-immobilized rats. Administration of fluoxetine-HCl (20 mg/kg) inhibited holo and total Tp pyrrolase enzyme activities when compared to saline injected rats and ultimately increases liver Tp concentrations. Although serum total Tp remained unchanged but brain Tp and 5-HT concentrations were significantly increased. It is therefore concluded that pre-treatment of rats with fluoxetine-HCl did not alter conditioned stress induced change in brain Tp metabolism. Future studies on chronic administration of the drug will be fruitful to investigate its effects on Tp metabolism in conditioned stress. PMID- 16380346 TI - Pharmacological activity of morpholino compound. AB - The pharmacological effect of morpholino compound in different biological field is of great importance for researchers and investigators. They are working day and night to synthesize pharmacologically more effective morpholino derivatives with less toxic effects. This kind of pharmacological screening of synthesized compounds can lead to the discovery of biologically useful compounds. The findings of the present study are encouraging, since present compound showed a promising analgesic activities and anti-inflammatory. Chemically synthesized morpholino compound are tested for its analgesic and inflammatory activities in intact albino mice ad rat. Compound was administered orally in different doses using the tail immersion test in mice for analgesic activity. The results of this study demonstrated that compound has inhibitory effect on arachidonic acid metabolisms via cyclooxygenase pathway or we can say the inhibition of prostaglandins biosynthesis. Classical non-steroidal anti-inflammatory drug, which inhibits cyclooxygenase, can shift arachidonic metabolism to lipoxygenase products. These findings reflect a new fact of pharmacological action of this compound like inhibition of arachidonic acid pathway in vivo. It can be concluded that the present study revealed a fair success rate but it is necessary to carry out further investigation to confirm the results. PMID- 16380347 TI - In vitro availability of lomefloxacin hydrochloride in presence of essential and trace elements. AB - Lomefloxacin hydrochloride is a third generation fluoroquinolone antibacterial agent having a broad spectrum of action against a wide range of gram-positive and gram-negative organisms. The in vitro availability studies of lomefloxacin were carried out in presence of essential and trace elements such as magnesium, calcium, chromium, ferric, ferrous, cobalt, nickel, copper, zinc and cadmium in simulated gastric juice, simulated intestinal juice and blood pH at 37 degrees C using B.P 2003 dissolution test apparatus. It was observed that availability of lomefloxacin was depressed in presence of nickel and zinc in simulated gastric juice and in presence of Fe2+ in simulated intestinal juice, while many metals like magnesium, chromium, iron (both Fe2+ and Fe3+), cobalt, nickel, copper and cadmium depressed the availability of lomefloxacin at blood pH. Furthermore, the availability of lomefloxacin alone in simulated intestinal juice and at blood pH was reduced as compared to simulated gastric juice. The antibacterial activities of lomefloxacin in presence of these metal ions were observed and compared to control against six different microorganisms i.e., Staphylococcus aureus, Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Bacillus fragilis and Streptococcus pneumonia by disc diffusion method to measure the inhibitory zone and MIC were determined by tube dilution method. PMID- 16380348 TI - Computer aided conformational analysis of sulfonated azo dyes diammonium orange G (C16H10N2O7S2 (NH4) x 4H2O. AB - The present work describes the conformational analysis of diammonium orange G (C16H10N2O7S2 (NH4) x 4H2O) by using Kitaigorodsky function. The minimum potential energy was found to be -0.0099839 at omega1 = 16 degrees and omega2 = 360 degrees. PMID- 16380349 TI - Relation of serum sialic acid with serum lipids in cardiac patients. AB - Elevated serum sialic acid concentration is a risk factor for coronary heart disease in general population along with dyslipidemia. Sialic acid and lipid profile was determined in healthy subjects (Group A1 and A2) and sixty cardiac patients, divided into two groups, group B comprising of thirty male patients and group C comprising of thirty female patients. A highly significant increase was observed in the level of sialic acid and different variables of lipid profile except HDL-C in all patients. HDL-C showed a significant decrease in patients. An increase in the level of sialic acid in cardiac patients showing the implications of raised level of sialic acid in the development of cardiovascular complications. PMID- 16380350 TI - Spectrophotometric methods for the determination of Ibuprofen in tablets. AB - Ibuprofen in film coated tablets of different strengths has been determined using different spectrophotometric methods. These are: (i) the compensation method, (ii) a two wavelengths method, (iii) second-order and (iv) fourth-order derivative methods, and (v) a curve fitting method based upon computing the quadratic coefficient of the orthogonal polynomials expansion of its benzenoid absorption characteristics. All results were compared with the HPLC method of the B.P. 2003 using paired comparison. Developed methods have been validated and applied to different tablet formulations. Mean differences from the B.P. method were found to be -0.10, -0.30, -0.10, -0.16, -0.34%, respectively. In view of the relatively low specific absorbance of ibuprofen in the ultraviolet region [A (1%, 1 cm) =18.5 at 264 nm] its accurate and precise determination in different tablet formulations is challenging due to the presence of interferences from excipients. PMID- 16380351 TI - In vitro availability of trimethylphloroglucinol and its degradation product from dosage formulations by RP-HPLC. AB - A simple, sensitive, reliable, and rapid high-performance liquid chromatographic method for the simultaneous determination of phloroglucinol and trimethyl phloroglucinol has been developed. Acetonitrile-water (1:1 v/v) was used as mobile phase, with flow rate 2 ml/minutes. pH was adjusted to 3 with phosphoric acid. U.V detection was performed at 242 nm. The results obtained showed a good agreement with the declared content. Recovery values for phloroglucinol were from 99.91% to 100.62% and recovery values for trimethylphloroglucinol were from 98.44% to 100.04%. The proposed method is rapid, accurate, and selective, it may be used for the quantitative analysis of phloroglucinol and trimethylphloroglucinol. The method was found to be specific, accurate, precise and reliable for the determination of phloroglucinol and trimethylphloroglucinol in the form of raw materials, in bulk drugs and formulation. It was possible to determine both phloroglucinol and trimethylphloroglucinol in the concentration range of 5 nano gram to 30 nano grams. The detection limit of both phloroglucinol and trimethylphloroglucinol were 0.4 nano gram. PMID- 16380352 TI - Anti-inflammatory, analgesic and diuretic activity of Polygonum lanatum Roxb. AB - The hexane (PLH), ethyl acetate (PLE) and methanol (PLM) extracts of dried whole plant parts of Polygonum lanatum Roxb. (Family, Polygonaceae) obtained by successive cold extraction, were subjected to evaluate anti-inflammatory, analgesic and diuretic activity in experimental animals. Oral administration of either PLH and PLM at a dose of 300 mg/kg body weight showed statistically significant (p < 0.001) inhibition of rat paw edema by 41.09% and 30.15%, respectively, which was comparable to that of standard drug phenylbutazone (42.15%). Compared to the inhibition of acetic acid-induced writhing by aminopyrine (69.94%, p < 0.001), treatment with either PLH, PLE or PLM elicited significant inhibition of acetic acid-induced writhing reflex by 44.80% (p < 0.001), 33.87% (p < 0.01) and 62.29% (p < 0.001), respectively. In addition, mild to potent diuretic activity was observed after oral administration of these extracts in Swiss albino mice. PMID- 16380353 TI - Resistance pattern of antibiotics against clinical isolates of Staphylococcus aureus. AB - The present research was conducted to study the susceptibility and resistance pattern of Staphylococcus aureus isolates against various brands of commonly used antibiotics. A total of 50 bacterial isolates were obtained from various clinical specimens submitted to the clinical laboratory of Pakistan Institute of Medical Sciences Islamabad (PIMS). Out of 50 isolates 30 (60%) were identified as coagulase positive Staphylococcus aureus. These isolates were tested for susceptibility pattern 19 (63%), these isolates were resistant to penicillin, 17 (57%) to ampicillin, 19 (63%) to cloxacillin, 20 (67%) to streptomycin, 11 (37%) to clindamycin, 16 (53%) to lincomycin and 19 (63%) to neomycin. These isolates had MIC range of penicillin and lincomycin (<0.5 to > 512 microg/mL), ampicillin, cloxacillin and clindamycin (0.5 to 256 microg/mL), streptomycin (4 to >512 microg/mL) and neomycin (2 to 256 microg/mL). Clindamycin was found to be the most effective among these antibiotics tested in the study against these clinical isolates. PMID- 16380354 TI - In vitro monitoring of ciprofloxacin antacids interactions by UV & HPLC. AB - Ciprofloxacin or 1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-7-(1-piperazinyl]-3 quinoline carboxylic acid is a fluorinated quinolone antibacterial agent extensively used worldwide. There are number of drug interactions reported for this antibacterial agent. In present studies, in vitro release of ciprofloxacin hydrochloride in presence of various antacids like sodium bicarbonate, calcium hydroxide, calcium carbonate, aluminum hydroxide, magnesium hydroxide, magnesium carbonate, magnesium trisilicate and magaldrate has been studied on BP 2002 dissolution test apparatus. Drug in each case was analyzed either by measuring the absorbance of aliquots at 278 and 316 nm on a UV/VIS spectrophotometer, or by reversed-phase high-performance liquid chromatographic (RP-HPLC) method. These studies were carried out in simulated gastric and intestinal juices for three hours at 37 degrees C. The availability of ciprofloxacin was found to be markedly retarded in presence of all the antacids studied. PMID- 16380355 TI - Phytochemical and active column fractions of Pyrenacantha staudtii leaf extracts on isolated rat uterus. AB - Four methanolic leaf fraction extracts of Pyrenacantha staudtii obtained from accelerated gradient chromatography (AGC) were tested on the isolated rat uterus. Various fractions--M1, M2, M3 and M4 through bioassay guided isolation were obtained. Fractions M(2) and M(4), containing saponins and alkaloids respectively, significantly (P<0.05) exerted high smooth muscle relaxant activity on the uterus. Fractions M(1) and M(3) containing fatty acids and tannins respectively did not exhibit significant effect on the isolated rat uterus. The results indicate the presence of active principles in the leaf extracts of P. staudtii which may be responsible for some of the applications in traditional medicines as remedy against threatened abortion and dysmenorrheal. PMID- 16380356 TI - Synthesis and characterization of gliclazide complexes of magnesium, calcium, chromium, manganese, iron, nickel, copper, zinc and cadmium salts. AB - Gliclazide is a well known agent used for NIDDM. Present paper reports the synthesis and characterization of its metal complexes with magnesium, calcium, chromium, manganese, iron, nickel, copper, zinc and cadmium. These complexes were characterized through physical characteristics, IR, H(1)-NMR, and atomic absorption spectroscopic studies. PMID- 16380357 TI - The effects of mefenamic acid on the blood haemoglobin of the lizard, Uromastix hardwickii. AB - This study deals with the effect of 7.1 mg/day, 10.5 mg/day and 14 mg/day doses of mefenamic acid administered for 12 days to three groups of Uromastix hardwickii respectively. Individual blood samples were obtained from the anterior abdominal vein and hemoglobin content was determined. The hemoglobin in test was 5.1 g/100 ml compared to 8.0 g/100 ml of controls in experiment I and its amount remained almost similar in the case of experiment II, whereas, 4.5 g/100 ml was observed of test compared to 8.0 g/100 ml of their counterparts. PMID- 16380358 TI - Grape fruit juice-drug interactions. AB - Grapefruit juice can markedly augment oral drug bioavailability was originally based on an unexpected observation from an interaction study between the dihydropyridine calcium channel antagonist, felodipine, and ethanol in which grapefruit juice was used to mask the taste of the ethanol. Subsequent investigations showed that grapefruit juice acted by reducing presystemic felodipine metabolism through selective post-translational down regulation of cytochrome P450 3A4 (CYP3A4) expression in the intestinal wall. Since the duration of effect of grapefruit juice can last 24h, repeated juice consumption can result in a cumulative increase in felodipine AUC and C(max). Clinically relevant interactions seem likely for most dihydropyridines, terfenadine, saquinavir, cyclosporin, midazolam, triazolam and verapamil and may also occur with lovastatin, cisapride and astemizole (Guo et al., 2000). The high variability of the magnitude of effect among individuals appeared dependent upon inherent differences in enteric CYP3A4 protein expression such that individuals with highest baseline CYP3A4 had the highest proportional increase. At least 20 other drugs have been assessed for an interaction with grapefruit juice metabolism mediated by CYP3A4 appear affected by grapefruit juice. Clinically relevant interactions seem likely for most dihydropyridines, terfenadine, saquinavir, cyclosporin, midazolam, triazolam and verapamil and may also occur with lovastatin, cisapride and astemizole. The importance of these interactions appears to be influenced by individual patient susceptibility, type and amount of grapefruit juice and administration-related factors. Although in vitro findings support the flavonoid, naringin, or the furanocoumarin, 6', 7' dihydroxybergamottin, as being active ingredients. A recent investigation indicated that neither of these substances made a major contribution to grapefruit juice-drug interactions in humans (Guo et al., 2000). PMID- 16380359 TI - Effect of magnesium and zinc on antimicrobial activities of some antibiotics. AB - Mg and Zn are essential elements in biological system. They are essential for enzymatic activity, maintaining three-dimensional structure of proteins, for the synthesis of nucleic acids and proteins etc. Deficiency of Mg and Zn causes different life threatening diseases. That is why the optimum level of Mg and Zn must be maintained for all biological systems. The experiment was aimed to evaluate the effect of magnesium and zinc on the efficacy of antibiotic agents against different microorganisms. It has been observed that the antibiotic activity of an antibiotic agent increased significantly with concomitant use of Mg salt at a concentration ranging from 3-9 microg per antibiotic disc. Similarly, Zn salt increased the activity of an antibiotic at a concentration ranging from 9-15 microg per antibiotic disc. The experiment revealed that concomitant administration of antibiotic with Mg or Zn salt not only will supplement the deficiency of these electrolytes but also will increase the activity of antibiotics against different bacterial strains. PMID- 16380360 TI - Effects of pefloxacin in multi drug resistant typhoid Fever. AB - In 28 children, with bacteriologically and/or serologically diagnosed typhoid fever treated at CMH, Rawalpindi in 2003, first one of the three recommended drugs (viz. chloramphenicol, amoxycillin or co-trimoxazole) was given for 7 days for defervescence to occur. In those who failed to respond a second trial of therapy with one of the other two drugs was initiated, after excluding the first drug. A second failure of therapy was taken as an indication to use pefloxacin singly. Finally, 18 (64.3%) cases responded to chloramphenicol or amoxycillin or co-trimoxazole. Pefloxacin was used in 10 (35.7%) cases. The failure rate of treatment with chloramphenicol was 50%, with amoxycillin 71.4% with co trimoxazole 75% and 0% with pefloxacin. An analysis of the 28 cases revealed that apart from fever (in 100%), splenomegaly (in 82.1%) was the most important clinical indicator to diagnosis. along with absolute eosinopenia (in 71.4%). There were no major complications, except 2 cases with typhoid hepatitis that responded to choramphenicol and co-trimoxazole, respectively. Blood culture grew Salmonella typhi in 7 cases of which 5 (72%) were multi drug resistant S. typhi. PMID- 16380361 TI - Saving Sex for Later: an evaluation of a parent education intervention. AB - CONTEXT: Initiation of sexual intercourse prior to high school is prevalent among inner-city black and Hispanic youths, and has multiple negative health and social consequences. A promising strategy for addressing early adolescent sexual activity is parent education that addresses normal pubertal changes and the challenges of becoming a teenager. METHODS: A 2003-2005 randomized trial to test the effectiveness of Saving Sex for Later, a parent education program presented on three audio CDs, enrolled 846 families with fifth- and sixth-grade students in seven New York City schools. Parent and youth surveys were conducted at baseline and three months postintervention. Multivariate logistic and linear regression analyses were performed to assess relationships between youth and parent outcomes and treatment condition. RESULTS: At follow-up, parents in the intervention group were significantly more likely than controls to score high on indexes of communication with children about targeted risk behaviors, self-efficacy to discuss pubertal development and sexuality, and perceived influence over youths' behaviors (odds ratios, 1.9-2.5). Youths in the intervention condition were more likely than controls to report high family support, and reported more family rules and fewer behavioral risks. Family support and rules partially mediate the relationship between treatment condition and behavioral risks. CONCLUSION: Saving Sex for Later is a promising intervention for promoting youths' sexual abstinence. The intervention may also be effective in enhancing positive parenting practices among parents who are typically difficult to reach because of economic hardship, full schedules and complicated lives. PMID- 16380362 TI - Social-cognitive predictors of consistent condom use among young people in Moscow. AB - CONTEXT: Russia is experiencing an explosive HIV epidemic, and young people aged 15-29 have the highest incidence of infection. Understanding factors associated with condom use in this age-group is important in developing effective prevention interventions. METHODS: Telephone interviews were conducted with 1,203 Muscovites aged 15-29 in September 2002 to assess condom use, HIV knowledge and sexual behavior. Multivariate logistic regression was used to determine independent predictors of consistent condom use. RESULTS: Forty-four percent of sexually experienced respondents reported using condoms consistently. In multivariate analysis, the likelihood of consistent use was elevated among single women and men (odds ratios, 1.8 and 2.6, respectively), those who considered condoms reliable protection against unwanted pregnancy (2.4 and 1.6) and those who believed that most of their peers use condoms (2.9 and 4.6). For women, having recently had multiple partners was associated with increased odds of consistent use, and consistent use declined with age. CONCLUSIONS: Strategies to promote condom use should increase awareness about their effectiveness against not only unwanted pregnancies but also HIV and other STDs. Condoms should be recommended for married couples and people with one permanent partner as a contraceptive option as well as for disease prevention. PMID- 16380363 TI - Pregnancy planning status and health behaviors among nonpregnant women in a California managed health care organization. AB - CONTEXT: Women's behaviors before and during pregnancy can affect their infants' health. Particularly because many births in the United States are unintended, it is important to understand women's health behaviors and pregnancy planning status before they become pregnant. METHODS: A telephone survey of nonpregnant women of childbearing age who belonged to a Southern California managed care plan was conducted from 1998 through 2000. Survey data were analyzed in logistic regression models assessing differences in selected behaviors between women planning pregnancy and others. RESULTS: Compared with women not planning pregnancy, those planning pregnancy within the next year ("soon") were less likely to report smoking (odds ratio, 0.6), and more likely to report taking a multivitamin regularly (1.4) and having had a health care visit in the past year (1.6). Women planning a pregnancy more than one year in the future had elevated odds of reporting alcohol use (1.4); they were similar to women not planning pregnancy with respect to multivitamin use and smoking behavior. Women planning pregnancy soon were more likely than women not planning pregnancy to report that a health care professional had talked to them about taking a vitamin or mineral supplement (1.6). CONCLUSIONS: All women of childbearing age need information about the importance of engaging in healthy behaviors. Health care providers who have regular contact with such women should send clear messages about the adverse effects of alcohol and smoking during pregnancy and the importance of taking a multivitamin regularly, regardless of women's pregnancy plans, before they become pregnant. PMID- 16380364 TI - Consistency of condom use among low-income hormonal contraceptive users. AB - CONTEXT: Hormonal contraceptive users may be at increased risk for HIV and other STDs. An understanding of their decisions and abilities to use condoms is needed to focus intervention programs aimed at improving their protective behaviors. METHODS: Between 1999 and 2001, 426 new users of depot medroxyprogesterone acetate (DMPA) and oral contraceptives were recruited from public clinics providing family planning services to low-income women and surveyed when they began their method and again three months later. Bivariate analyses examined the consistency of condom use across subgroups, and multivariate analyses assessed associations between consistent use and various characteristics. RESULTS: Among women who had used condoms consistently before starting on DMPA or the pill, 54% discontinued consistent use after taking these contraceptives. Overall, 20% of women consistently used condoms with their hormonal method, and such use did not vary significantly by contraceptive type. Seventy-five percent of women in non monogamous relationships were inconsistent users, though nearly a third had been consistent users prior to beginning a hormonal method. Factors associated with an elevated likelihood of consistent use were the male partner's positive opinion of condoms (odds ratio, 3.3) and the woman's strong belief that condom use is important for vaginal intercourse (3.5) and even if the couple is using another form of birth control (4.1). CONCLUSIONS: Many women at highest risk for disease have a decreased likelihood of using condoms, and disease prevention programs should be customized to target these women. Educational efforts focusing on women's attitudes and negotiation skills may be the best means of increasing dual method use. PMID- 16380365 TI - Parent-child relations among minor females attending U.S. family planning clinics. AB - CONTEXT: Relatively little is known about parent-child relations among minor females who use family planning clinics. Such information could inform the debate on parental involvement legislation and help clinics develop effective strategies to promote positive parental engagement. METHODS: Self-administered surveys were completed in 2003-2004 by 1,526 women younger than 18 attending 79 U.S. family planning clinics, providing measures of parent-child relations, perceived parental attitudes toward sex and birth control, and parental knowledge of the clinic visit. Associations between relationships with parents and parental knowledge of clinic visits were examined using t tests and logistic regression. RESULTS: Many adolescents had talked to parents about sexual issues (50-80%, depending on the topic) and reported high levels of connectedness with parents (68%). A substantial minority (19%) perceived that parents disapprove of their both having sex and using birth control. The majority (60%) reported that a parent knew of their clinic visit; such reports were most common among those who had high levels of connectedness to parents and communication with parents about sexual issues, and those who did not perceive parents to disapprove of sex and birth control. Adolescents aged 15 and younger were more likely than 17-year-olds to indicate that a parent knew they were at the clinic and to report that a parent suggested the clinic. CONCLUSIONS: Overall, minors attending family planning clinics have good relations with parents. The youngest adolescents may be at family planning clinics specifically because parents are involved in their reproductive health decisions. PMID- 16380366 TI - Changing the face of long-term care. AB - Long-term care (LTC) for older persons is based on the wrong foundations. It is too wedded to protection and not directed enough to maximizing personal goals. Tinkering at the margins will not suffice. Bigger, bolder steps are needed. A first step is to re-examine the goals of such care and the tools we have at hand to meet them. We need to re-examine the major elements of what we have taken for granted. A number of forces must be harnessed, and in some cases effort must be redistributed. Effective collaboration will depend on shared goals. Unless LTC is viewed as something that is desired, we will never receive more than reluctant support from any quarter. Good care must be seen as making a difference; that difference can be in function or in quality of life; it may be as subtle as slowing decline, but it must be made apparent to be appreciated. PMID- 16380367 TI - Chain reaction: an exploratory study of nursing home bankruptcy in California. AB - This paper reports on an exploratory study of nursing home bankruptcy. From state and industry data regarding nearly 1,000 California facilities, it was possible to identify 155 homes in five chains (multi-facility organizations) that were operating in bankruptcy in 2000. When compared with facilities in non-bankrupt chains, while the bankrupt chain facilities had significantly worse financial liquidity, higher administrative costs, and higher payables to related parties, they also had more Medicare residents, fewer Medicaid residents, better solvency, and were located in less competitive county markets and in areas with higher Medicaid reimbursement rates. These findings indicate that, rather than facility characteristics and local market factors, strategic decisions taken at the corporate (chain) level are the major determinants of nursing facility bankruptcy status. PMID- 16380368 TI - Emerging services for community-based long-term care in urban China: a systematic analysis of Shanghai's community-based agencies. AB - China's rapid economic reforms, coupled with the changes in age composition of the demographic structure, have greatly affected the traditional family support system. In response to these changes, efforts to develop new models of community based long-term care (CBLTC) for elders in China have received growing attention. This paper provides a systematic analysis of the current status of emerging CBLTC systems in Shanghai, China. It covers several domains of the system: service delivery, workforce, financing, and quality of care management. Several main issues involved in the development of the emerging system are addressed, and relevant policy implications are presented in the paper. PMID- 16380369 TI - A comparison of early retirement pensions in the United States and Russia: the pensions of musicians. AB - Early retirement pensions for particular occupations free national policy to establish the social security early retirement age at a later age that is more appropriate for the population as a whole. This paper focuses on early retirement pensions in the United States and the Russian Federation. While comparing early retirement pensions generally, the paper provides a more detailed discussion of the pensions for musicians. While this is an unconventional group to choose for the study of pensions, study of their pensions yields insights into the principles underlying retirement age policy in the two countries. PMID- 16380370 TI - An international review of the long-term care workforce: policies and shortages. AB - The developed world's population is aging, due to trends of increased life expectancies and decreased fertility rates. These trends are predicted to increase demand on long-term care services. At the same time, the long-term care workforce is in shortage in most of the developed world. Moreover, such shortages are expected to increase due to parallel socio-demographic factors. The increase in demand for longterm care, coupled with shortage in supply of care workers, has promoted some attention from policymakers. The current paper provides an international review of institutional arrangements for long-term care in different developed countries and in particular explores different strategies used or proposed to resolve the shortage in the long-term care workforce. PMID- 16380371 TI - CDP-choline significantly restores phosphatidylcholine levels by differentially affecting phospholipase A2 and CTP: phosphocholine cytidylyltransferase after stroke. AB - Phosphatidylcholine (PtdCho) is a major membrane phospholipid, and its loss is sufficient in itself to induce cell death. PtdCho homeostasis is regulated by the balance between hydrolysis and synthesis. PtdCho is hydrolyzed by phospholipase A2 (PLA2), PtdChospecific phospholipase C (PtdCho-PLC), and phospholipase D (PLD). PtdCho synthesis is rate-limited by CTP:phosphocholine cytidylyltransferase (CCT), which makes CDP-choline. The final step of PtdCho synthesis is catalyzed by CDP-choline:1,2-diacylglycerol cholinephosphotransferase. PtdCho synthesis in the brain is predominantly through the CDP-choline pathway. Transient middle cerebral artery occlusion (tMCAO) significantly increased PLA2 activity, secretory PLA2 (sPLA2)-IIA mRNA and protein levels, PtdCho-PLC activity, and PLD2 protein expression following reperfusion. CDP-choline treatment significantly attenuated PLA2 activity, sPLA2 IIA mRNA and protein levels, and PtdCho-PLC activity, but did not affect PLD2 protein expression. tMCAO also resulted in loss of CCT activity and CCTalpha protein, which were partially restored by CDP-choline. No changes were observed in cytosolic PLA2 or calcium-independent PLA2 tMCAO. protein levels after Up regulation of PLA2, PtdCho-PLC, and PLD and regulation of CCT collectively down resulted in loss of PtdCho, which was significantly restored by CDP-choline treatment. CDP-choline treatment significantly attenuated the infarction volume by 55 +/- 5% after 1 h of tMCAO and 1 day of reperfusion. Taken together, these results suggest that CDP-choline significantly restores Ptd-Cho levels by differentially affecting sPLA2-IIA, PtdCho-PLC, and CCTalpha after transient focal cerebral ischemia. A hypothetical scheme is proposed integrating results from this study and from other reports in the literature. PMID- 16380372 TI - Characterization of human DHRS6, an orphan short chain dehydrogenase/reductase enzyme: a novel, cytosolic type 2 R-beta-hydroxybutyrate dehydrogenase. AB - Human DHRS6 is a previously uncharacterized member of the short chain dehydrogenases/reductase family and displays significant homologies to bacterial hydroxybutyrate dehydrogenases. Substrate screening reveals sole NAD(+)-dependent conversion of (R)-hydroxybutyrate to acetoacetate with K(m) values of about 10 mm, consistent with plasma levels of circulating ketone bodies in situations of starvation or ketoacidosis. The structure of human DHRS6 was determined at a resolution of 1.8 A in complex with NAD(H) and reveals a tetrameric organization with a short chain dehydrogenases/reductase-typical folding pattern. A highly conserved triad of Arg residues ("triple R" motif consisting of Arg(144), Arg(188), and Arg(205)) was found to bind a sulfate molecule at the active site. Docking analysis of R-beta-hydroxybutyrate into the active site reveals an experimentally consistent model of substrate carboxylate binding and catalytically competent orientation. GFP reporter gene analysis reveals a cytosolic localization upon transfection into mammalian cells. These data establish DHRS6 as a novel, cytosolic type 2 (R)-hydroxybutyrate dehydrogenase, distinct from its well characterized mitochondrial type 1 counterpart. The properties determined for DHRS6 suggest a possible physiological role in cytosolic ketone body utilization, either as a secondary system for energy supply in starvation or to generate precursors for lipid and sterol synthesis. PMID- 16380373 TI - Sec14 homology domain targets p50RhoGAP to endosomes and provides a link between Rab and Rho GTPases. AB - Sec14 protein was first identified in Saccharomyces cerevisiae, where it serves as a phosphatidylinositol transfer protein that is essential for the transport of secretory proteins from the Golgi complex. A protein domain homologous to Sec14 was identified in several mammalian proteins that regulates Rho GTPases, including exchange factors and GTPase activating proteins. P50RhoGAP, the first identified GTPase activating protein for Rho GTPases, is composed of a Sec14-like domain and a Rho-GTPase activating protein (GAP) domain. The biological function of its Sec14-like domain is still unknown. Here we show that p50RhoGAP is present on endosomal membranes, where it colocalizes with internalized transferrin receptor. We demonstrate that the Sec14-like domain of P50RhoGAP is responsible for the endosomal targeting of the protein. We also show that overexpression of p50RhoGAP or its Sec14-like domain inhibits transferrin uptake. Furthermore, both P50RhoGAP and its Sec14-like domain show colocalization with small GTPases Rab11 and Rab5. We measured bioluminescence resonance energy transfer between p50RhoGAP and Rab11, indicating that these proteins form molecular complex in vivo on endosomal membranes. The interaction was mediated by the Sec 14-like domain of p50RhoGAP. Our results indicate that Sec14-like domain, which was previously considered as a phospholipid binding module, may have a role in the mediation of protein-protein interactions. We suggest that p50RhoGAP provides a link between Rab and Rho GTPases in the regulation of receptor-mediated endocytosis. PMID- 16380374 TI - Cholesterol-induced apoptotic macrophages elicit an inflammatory response in phagocytes, which is partially attenuated by the Mer receptor. AB - Macrophage apoptosis and the ability of phagocytes to clear these apoptotic cells are important processes in advanced atherosclerosis. Phagocytic clearance not only disposes of dead cells but usually elicits an anti-inflammatory response. To study this process in a model of advanced lesional macrophage death, macrophages rendered apoptotic by free cholesterol loading (FC-AMs) were incubated briefly with fresh macrophages ("phagocytes"). FC-AMs were promptly ingested by the phagocytes, which was dependent upon actin polymerization and the phagocyte Mer receptor. Surprisingly, this brief exposure to FC-AMs triggered a modest proinflammatory response in the phagocytes: tumor necrosis factor-alpha (TNF alpha) and interleukin (IL)-1beta were induced, whereas the levels of transforming growth factor-beta and IL-10 were not increased. This response required cell contact between the FC-AMs and phagocytes but not FC-AM ingestion. TNF-alpha and IL-1beta induction required one or more proteins on the FC-AM surface and was dependent on signaling through extracellular signal-regulated kinase-1/2 mitogen-activated protein kinase and nuclear factor-kappaB in the phagocytes. TNF-alpha production was markedly greater when Mer-defective phagocytes were used, indicating that Mer attenuated the inflammatory response. Interestingly, a more typical anti-inflammatory response was elicited when phagocytes were exposed to macrophages rendered apoptotic by oxidized low density lipoprotein or UV radiation. Thus, the proinflammatory milieu of advanced atherosclerotic lesions may be promoted, or at least not dampened, by contact between FC-induced apoptotic macrophages and neighboring phagocytes prior to apoptotic cell ingestion. PMID- 16380375 TI - Inhibition of Werner syndrome helicase activity by benzo[a]pyrene diol epoxide adducts can be overcome by replication protein A. AB - RecQ helicases are believed to function in repairing replication forks stalled by DNA damage and may also play a role in the intra-S-phase checkpoint, which delays the replication of damaged DNA, thus permitting repair to occur. Since little is known regarding the effects of DNA damage on RecQ helicases, and because the replication and recombination defects in Werner syndrome cells may reflect abnormal processing of damaged DNA associated with the replication fork, we examined the effects of specific bulky, covalent adducts at N(6) of deoxyadenosine (dA) or N(2) of deoxyguanosine (dG) on Werner (WRN) syndrome helicase activity. The adducts are derived from the optically active 7,8-diol 9,10-epoxide (DE) metabolites of the carcinogen benzo[a]pyrene (BaP). The results demonstrate that WRN helicase activity is inhibited in a strand-specific manner by BaP DE-dG adducts only when on the translocating strand. These adducts either occupy the minor groove without significant perturbation of DNA structure (trans adducts) or cause base displacement at the adduct site (cis adducts). In contrast, helicase activity is only mildly affected by intercalating BaP DE-dA adducts that locally perturb DNA double helical structure. This differs from our previous observation that intercalating dA adducts derived from benzo[c]phenanthrene (BcPh) DEs inhibit WRN activity in a strand- and stereospecific manner. Partial unwinding of the DNA helix at BaP DE-dA adduct sites may make such adducted DNAs more susceptible to the action of helicase than DNA containing the corresponding BcPh DE-dA adducts, which cause little or no destabilization of duplex DNA. The single-stranded DNA binding protein RPA, an auxiliary factor for WRN helicase, enabled the DNA unwinding enzyme to overcome inhibition by either the trans-R or cis-R BaP DE-dG adduct, suggesting that WRN and RPA may function together to unwind duplex DNA harboring specific covalent adducts that otherwise block WRN helicase acting alone. PMID- 16380376 TI - Profiling of early gene expression induced by erythropoietin receptor structural variants. AB - The development of erythroid progenitor cells is triggered via the expression of the erythropoietin receptor (EPOR) and its activation by erythropoietin. The function of the resulting receptor complex depends critically on the presence of activated JAK2, and the complex contains a large number of signaling molecules recruited to eight phosphorylated tyrosine residues. Studies using mutant receptor forms have demonstrated that truncated receptors lacking all tyrosines are able to support red blood cell development with low efficiency, whereas add back mutants containing either Tyr343 or Tyr479 reconstitute EPOR signaling and erythropoiesis in vivo. To study the contribution of tyrosines to receptor function, we analyzed the activation of essential signaling pathways and early gene induction promoted by different receptor structural variants using human epidermal growth factor receptor/murine EPOR hybrids. In our experiments, receptors lacking all tyrosine residues or the JAK2-binding site did not induce mitogenic and anti-apoptotic signaling, whereas add-back mutant receptors containing single tyrosine residues (Try343 and Tyr479) supported the activation of these functions efficiently. Profiling of early gene expression using cDNA array hybridization revealed that (i) the high redundancy in the activation of signaling pathways is continued at the level of transcription; (ii) the expression of many genes targeted by the wild-type receptor is not supported by add-back mutants; and (iii) a small set of genes are exclusively induced by add back receptors. We report the identification of several early genes that have not been implicated in the EPOR-dependent response so far. PMID- 16380377 TI - The nuclear import of protein kinase D3 requires its catalytic activity. AB - The protein kinase D (PKD) family consists of three serine/threonine protein kinases termed PKD, PKD2, and PKD3, which are similar in overall structure and primary amino acid sequence. However, each isozyme displays a distinctive intracellular localization. Taking advantage of the structural homology and opposite nuclear localization of PKD2 and PKD3, we generated an extensive set of chimeric proteins between both isozymes to determine which PKD3 domain(s) mediates its nuclear localization. We found that the C-terminal region of PKD3, which contains its catalytic domain, is necessary but not sufficient for its nuclear localization. Real time imaging of a photoactivatable green fluorescent protein fused to PKD3 revealed that point mutations that render PKD3 catalytically inactive completely prevented its nuclear import despite its interaction with importin alpha and beta. We also found that activation loop phosphorylation of PKD3 did not require its nuclear localization, and it was not sufficient to promote the nuclear import of PKD3. These results identify a novel function for the kinase activity of PKD3 in promoting its nuclear entry and suggest that the catalytic activity of PKD3 may regulate its nuclear import through autophosphorylation and/or interaction with another protein(s). PMID- 16380378 TI - Genetic and biochemical properties of streptococcal NAD-glycohydrolase inhibitor. AB - The gene encoding streptolysin O (slo), a cytolysin of hemolytic streptococci, is transcribed polycistronically from the promoter of the preceding NAD glycohydrolase (NADase) gene (nga). Between nga and slo, a putative open reading frame (orf1) is located whose function has been totally unknown. Present investigation demonstrated that the orf1 encodes a protein designated as streptococcal NADase inhibitor (SNI). From its nucleotide sequence, SNI was inferred to comprise 161 amino acid residues and the deduced molecular weight was 18,800. This protein was detectable only within cells. Coexpression of SNI was essential for production of streptococcal NADase, and NADase precursor existed as an inactive complex with SNI, in recombinant Escherichia coli. Monomeric NADase and SNI rapidly formed in vitro a stable heterodimer complex in the ratio 1:1, resulting in complete suppression of the hydrolase activity. Unlike other bacterial NADase inhibitors, SNI was thermostable. This protein, coexpressed and complexed with NADase, may protect the producer cocci from exhaustion of NAD. PMID- 16380379 TI - Promoter organization of the interferon-A genes differentially affects virus induced expression and responsiveness to TBK1 and IKKepsilon. AB - Virus-induced expression of interferon (IFN)-A genes is regulated by two members of the IFN regulatory factor (IRF) family, IRF-3 and IRF-7, which are activated by phosphorylation during viral infection by the IKK-related serine/threonine kinases TBK1 and IkappaB kinase epsilon (IKKepsilon). In this study, we demonstrate that three IRF-binding sites located in the virus-responsive element mediate the transcriptional activation of the IFN-A4 promoter by IRF-3. The precise arrangement of these IRF elements is required for synergistic activation of the IFN-A4 promoter following Newcastle disease virus infection or activation by TBK1 or IKKepsilon. The ordered assembly of IRF-3 multimers on the promoter also determines cooperative recruitment of IRF-3 and CREB-binding protein and differential virus-induced expression of IFN-A4 gene promoter compared with IFN A11. Naturally occurring nucleotide substitutions disrupt two of the IRF elements in the IFN-A11 gene promoter, leading to a dramatic decrease in IRF-3 and CREB binding protein recruitment and in IRF-3-dependent transcription. Transcription of the IFN-A4 promoter by IRF-7 is mediated by two IRF elements; promoter mutants that carry a reversed IRF element retain the ability to respond to IKKepsilon or TBK1 expression in the presence of IRF-7 but lose the capacity to respond to virus or kinase-induced IRF-3. Interestingly, IKKepsilon or TBK1 stimulates the IRF-7-mediated transcription of IFN-A11, although at a lesser extent compared with IFN-A4. Our data indicate that virus-induced expression of IFN-A genes is dictated by the organization of IRF elements within the IFN-A promoters and that the differential IFN-A gene expression, based on the IRF-3 responsiveness, is partially compensated in the presence of IRF-7 when both factors are activated by IKKepsilon or TBK1. PMID- 16380380 TI - The receptor protein-tyrosine phosphatase PTPmu interacts with IQGAP1. AB - The receptor protein-tyrosine phosphatase PTPmu is a member of the Ig superfamily of cell adhesion molecules. The extracellular domain of PTPmu contains motifs commonly found in cell adhesion molecules. The intracellular domain of PTPmu contains two conserved catalytic domains, only the membrane-proximal domain has catalytic activity. The unique features of PTPmu make it an attractive molecule to transduce signals upon cell-cell contact. PTPmu has been shown to regulate cadherin-mediated cell adhesion, neurite outgrowth, and axon guidance. Protein kinase C is a component of the PTPmu signaling pathway utilized to regulate these events. To aid in the further characterization of PTPmu signaling pathways, we used a series of GST-PTPmu fusion proteins, including catalytically inactive and substrate trapping mutants, to identify PTPmu-interacting proteins. We identified IQGAP1, a known regulator of the Rho GTPases, Cdc42 and Rac1, as a novel PTPmu interacting protein. We show that this interaction is due to direct binding. In addition, we demonstrate that amino acid residues 765-958 of PTPmu, which include the juxtamembrane domain and 35 residues of the first phosphatase domain, mediate the binding to IQGAP1. Furthermore, we demonstrate that constitutively active Cdc42, and to a lesser extent Rac1, enhances the interaction of PTPmu and IQGAP1. These data indicate PTPmu may regulate Rho-GTPase-dependent functions of IQGAP1 and suggest that IQGAP1 is a component of the PTPmu signaling pathway. In support of this, we show that a peptide that competes IQGAP1 binding to Rho GTPases blocks PTPmu-mediated neurite outgrowth. PMID- 16380381 TI - Mcl-1 interacts with truncated Bid and inhibits its induction of cytochrome c release and its role in receptor-mediated apoptosis. AB - Engagement of death receptors such as tumor necrosis factor-R1 and Fas brings about the cleavage of cytosolic Bid to truncated Bid (tBid), which translocates to mitochondria to activate Bax/Bak, resulting in the release of cytochrome c. The mechanism underlying the activation, however, is not fully understood. Here, we have identified the anti-apoptotic Bcl-2 family member Mcl-1 as a potent tBid binding partner. Site-directed mutagenesis reveals that the Bcl-2 homology (BH)3 domain of tBid is essential for binding to Mcl-1, whereas all three BH domains (BH1, BH2, and BH3) of Mcl-1 are required for interaction with tBid. In vitro studies using isolated mitochondria and recombinant proteins demonstrate that Mcl 1 strongly inhibits tBid-induced cytochrome c release. In addition to its ability to interact directly with Bax and Bak, tBid also binds Mcl-1 and displaces Bak from the Mcl-1-Bak complex. Importantly, overexpression of Mcl-1 confers resistance to the induction of apoptosis by both TRAIL and tumor necrosis factor alpha in HeLa cells, whereas targeting Mcl-1 by RNA interference sensitizes HeLa cells to TRAIL-induced apoptosis. Therefore, our study demonstrates a novel regulation of tBid by Mcl-1 through protein-protein interaction in apoptotic signaling from death receptors to mitochondria. PMID- 16380382 TI - N-methyl-D-aspartate blocks activation of JNK and mitochondrial apoptotic pathway induced by potassium deprivation in cerebellar granule cells. AB - During the postnatal development of cerebellum, lack of excitatory innervation from the mossy fibers results in cerebellar granule cell (CGC) apoptosis during the migration of the cells toward the internal granule cell layer. Accordingly, CGCs die by apoptosis when cultured in physiological KCl concentrations (5 mm; K5), and they survive in the presence of depolarizing conditions such as high KCl concentration (25 mm; K25) or N-methyl-D-aspartate (NMDA). We have recently shown that NMDA is able to exert a long lasting neuroprotective effect when added to immature (2 days in vitro) CGC cultures by inhibition of caspase-3 activity. Here we show that NMDA- and K25-mediated neuroprotection is associated with an increase in the levels of Bcl-2, an inhibition of K5-mediated increase in Bax, and the inhibition of the release of apoptogenic factors from mitochondria such as Smac/DIABLO and cytochrome c. Moreover, we have shown that similar effects are observed when c-Jun N-terminal kinases (JNKs) are inhibited and that treatment of CGC cultures with NMDA blocks K5-mediated JNK activation. These results allow us to postulate that the inhibition of JNK-mediated release of apoptogenic factors from mitochondria is involved in the NMDA protection from K5-mediated apoptosis of CGCs. PMID- 16380384 TI - Overexpression of CYP2E1 in mitochondria sensitizes HepG2 cells to the toxicity caused by depletion of glutathione. AB - Induction of CYP2E1 by ethanol is one mechanism by which ethanol causes oxidative stress and alcohol liver disease. Although CYP2E1 is predominantly found in the endoplasmic reticulum, it is also located in rat hepatic mitochondria. In the current study, chronic alcohol consumption induced rat hepatic mitochondrial CYP2E1. To study the role of mitochondrial targeted CYP2E1 in generating oxidative stress and causing damage to mitochondria, HepG2 lines overexpressing CYP2E1 in mitochondria (mE10 and mE27 cells) were established by transfecting a plasmid containing human CYP2E1 cDNA lacking the hydrophobic endoplasmic reticulum targeting signal sequence into HepG2 cells followed by G418 selection. A 40-kDa catalytically active NH2-terminally truncated form of CYP2E1 (mtCYP2E1) was detected in the mitochondrial compartment in these cells by Western blot analysis. Cell death caused by depletion of GSH by buthionine sulfoximine (BSO) was increased in mE10 and mE27 cells as compared with cells transfected with empty vector (pCI-neo). Antioxidants were able to abolish the loss of cell viability. Increased levels of reactive oxygen species and mitochondrial 3 nitrotyrosine and 4-hydroxynonenal protein adducts and decreased mitochondrial aconitase activity and mitochondrial membrane potential were observed in mE10 and mE27 cells treated with BSO. The mitochondrial membrane stabilizer, cyclosporine A, was also able to protect these cells from BSO toxicity. These results revealed that CYP2E1 in the mitochondrial compartment could induce oxidative stress in the mitochondria, damage mitochondria membrane potential, and cause a loss of cell viability. The accumulation of CYP2E1 in hepatic mitochondria induced by ethanol consumption might play an important role in alcohol liver disease. PMID- 16380383 TI - Cytochrome P-450 4F18 is the leukotriene B4 omega-1/omega-2 hydroxylase in mouse polymorphonuclear leukocytes: identification as the functional orthologue of human polymorphonuclear leukocyte CYP4F3A in the down-regulation of responses to LTB4. AB - Leukotriene B(4) (LTB(4)) is a potent chemoattractant for polymorphonuclear leukocytes (PMN) and other cells. Human PMN inactivate LTB(4) by omega-oxidation catalyzed by cytochrome P-450 (CYP) 4F3A. The contribution of the enzymatic inactivation of LTB(4) by CYP4Fs to down-regulating functional responses of cells to LTB(4) is unknown. To elucidate the role of CYP4F-mediated inactivation of LTB(4) in terminating the responses of PMN to LTB(4) and to identify a target for future genetic studies in mice, we have identified the enzyme that catalyzes the omega-1 and omega-2 oxidation of LTB(4) in mouse myeloid cells as CYP4F18. As determined by mass spectrometry, this enzyme catalyzes the conversion of LTB(4) to 19-OH LTB(4) and to a lesser extent 18-OH LTB(4). Inhibition of CYP4F18 resulted in a marked increase in calcium flux and a 220% increase in the chemotactic response of mouse PMN to LTB(4). CYP4F18 expression was induced in bone marrow-derived dendritic cells by bacterial lipopolysaccharide, a ligand for TLR4, and by poly(I.C), a ligand for TLR3. However, when bone marrow-derived myeloid dendritic cells trafficked to popliteal lymph nodes from paw pads, the expression of CYP4F18 was down-regulated. The results identify CYP4F18 as a critical protein in the regulation of LTB(4) metabolism and functional responses in mouse PMN and identify it as the functional orthologue of human PMN CYP4F3A. PMID- 16380385 TI - Scavenger receptor class B type I (SR-BI) is involved in vitamin E transport across the enterocyte. AB - Although cellular uptake of vitamin E was initially described as a passive process, recent studies in the liver and brain have shown that SR-BI (scavenger receptor class B type I) is involved in this phenomenon. As SR-BI is expressed at high levels in the intestine, the present study addressed the involvement of SR BI in vitamin E trafficking across enterocytes. Apical uptake and efflux of the main dietary forms of vitamin E were examined using Caco-2 TC-7 cell monolayers as a model of human intestinal epithelium. (R,R,R)-gamma-tocopherol bioavailability was compared between wild-type mice and mice overexpressing SR-BI in the intestine. The effect of vitamin E on enterocyte SR-BI mRNA levels was measured by real-time quantitative reverse transcription-PCR. Concentration dependent curves for vitamin E uptake were similar for (R,R,R)-alpha-, (R,R,R) gamma-, and dl-alpha-tocopherol. (R,R,R)-alpha-tocopherol transport was dependent on incubation temperature, with a 60% reduction in absorption at 4 degrees C compared with 37 degrees C (p < 0.05). Vitamin E flux in enterocytes was directed from the apical to the basal side, with a relative 10-fold reduction in the transfer process when measured in the opposite direction (p < 0.05). Co incubation with cholesterol, gamma-tocopherol, or lutein significantly impaired alpha-tocopherol absorption. Anti-human SR-BI antibodies and BLT1 (a chemical inhibitor of lipid transport via SR-BI) blocked up to 80% of vitamin E uptake and up to 30% of apical vitamin E efflux (p < 0.05), and similar results were obtained for (R,R,R)-gamma-tocopherol. SR-BI mRNA levels were not significantly modified after a 24-h incubation of Caco-2 cells with vitamin E. Finally, (R,R,R) gamma-tocopherol bioavailability was 2.7-fold higher in mice overexpressing SR-BI than in wild-type mice (p < 0.05). The present data show for the first time that vitamin E intestinal absorption is, at least in part, mediated by SR-BI. PMID- 16380386 TI - Neutral ceramidase encoded by the Asah2 gene is essential for the intestinal degradation of sphingolipids. AB - Complex sphingolipids are abundant as eukaryotic cell membrane components, whereas their metabolites, in particular ceramide, sphingosine, and sphingosine 1 phosphate, are involved in diverse cell signaling processes. In mammals, degradation of ceramide by ceramidase yields sphingosine, which is phosphorylated by the action of sphingosine kinase to generate sphingosine 1-phosphate. Therefore, ceramidases are key enzymes in the regulation of the cellular levels of ceramide, sphingosine, and sphingosine 1-phosphate. To explore the physiological functions of a neutral ceramidase with diverse cellular locations, we disrupted the Asah2 gene in mice. Asah2 null mice have a normal life span and do not show obvious abnormalities or major alterations in total ceramide levels in tissues. The Asah2-encoded neutral ceramidase is highly expressed in the small intestine along the brush border, suggesting that the neutral ceramidase may be involved in a pathway for the digestion of dietary sphingolipids. Indeed, Asah2 null mice were deficient in the intestinal degradation of ceramide. Thus, the results indicate that the Asah2-encoded neutral ceramidase is a key enzyme for the catabolism of dietary sphingolipids and regulates the levels of bioactive sphingolipid metabolites in the intestinal tract. PMID- 16380387 TI - The protein phosphatase 2A phosphatase activator is a novel peptidyl-prolyl cis/trans-isomerase. AB - The protein phosphatase 2A (PP2A) phosphatase activator (PTPA) is an essential protein involved in the regulation of PP2A and the PP2A-like enzymes. In this study we demonstrate that PTPA and its yeast homologues Ypa1 and Ypa2 can induce a conformational change in some model substrates. Using these model substrates in different assays with and without helper proteases, this isomerase activity is similar to the isomerase activity of FKBP12, the human cyclophilin A, and one of its yeast homologs Cpr7 but dissimilar to the isomerase activity of Pin1. However, neither FKBP12 nor Cpr7 can reactivate the inactive form of PP2A. Therefore, PTPA belongs to a novel peptidyl-prolyl cis/trans-isomerase (PPIase) family. The PPIase activity of PTPA correlates with its activating activity since both are stimulated by the presence of Mg2+ATP, and a PTPA mutant (Delta208-213) with 400-fold less activity in the activation reaction of PP2A also showed almost no PPIase activity. The point mutant Asp205 --> Gly (in Ypa1) identified this amino acid as essential for both activities. Moreover, PTPA dissociates the inactive form from the complex with the PP2A methylesterase. Finally, Pro190 in the catalytic subunit of PP2A (PP2AC) could be identified as the target Pro isomerized by PTPA/Mg2+ATP since among the 14 Pro residues present in 12 synthesized peptides representing the microenvironments of these prolines in PP2AC, only Pro190 could be isomerized by PTPA/Mg2+ATP. This Pro190 is present in a predicted loop structure near the catalytic center of PP2AC and, if mutated into a Phe, the phosphatase is inactive and can no longer be activated by PTPA/Mg2+ATP. PMID- 16380388 TI - Selective loss of fine tuning of Gq/11 signaling by RGS2 protein exacerbates cardiomyocyte hypertrophy. AB - Alterations in cardiac G protein-mediated signaling, most prominently G(q/11) signaling, are centrally involved in hypertrophy and heart failure development. Several RGS proteins that can act as negative regulators of G protein signaling are expressed in the heart, but their functional roles are still poorly understood. RGS expression changes have been described in hypertrophic and failing hearts. In this study, we report a marked decrease in RGS2 (but not other major cardiac RGS proteins (RGS3-RGS5)) that occurs prior to hypertrophy development in different models with enhanced G(q/11) signaling (transgenic expression of activated Galpha(q)(*) and pressure overload due to aortic constriction). To assess functional consequences of selective down-regulation of endogenous RGS2, we identified targeting sequences for effective RGS2 RNA interference and used lipid-based transfection to achieve uptake of fluorescently labeled RGS2 small interfering RNA in >90% of neonatal and adult ventricular myocytes. Endogenous RGS2 expression was dose-dependently suppressed (up to 90%) with no major change in RGS3-RGS5. RGS2 knockdown increased phenylephrine- and endothelin-1-induced phospholipase Cbeta stimulation in both cell types and exacerbated the hypertrophic effect (increase in cell size and radiolabeled protein) in neonatal myocytes, with no major change in G(q/11)-mediated ERK1/2, p38, or JNK activation. Taken together, this study demonstrates that endogenous RGS2 exerts functionally important inhibitory restraint on G(q/11)-mediated phospholipase Cbeta activation and hypertrophy in ventricular myocytes. Our findings point toward a potential pathophysiological role of loss of fine tuning due to selective RGS2 down-regulation in G(q/11)-mediated remodeling. Furthermore, this study shows the feasibility of effective RNA interference in cardiomyocytes using lipid-based small interfering RNA transfection. PMID- 16380389 TI - Muscle type-specific fatty acid metabolism in insulin resistance: an integrated in vivo study in Zucker diabetic fatty rats. AB - Intramyocellular lipid content (IMCL) serves as a good biomarker of skeletal muscle insulin resistance (IR). However, intracellular fatty acid metabolites [malonyl-CoA, long-chain acyl-CoA (LCACoA)] rather than IMCL are considered to be responsible for IR. This study aimed to investigate dynamics of IMCL and fatty acid metabolites during fed-to-starved-to-refed transition in lean and obese (IR) Zucker diabetic fatty rats in the following different muscle types: soleus (oxidative), extensor digitorum longus (EDL, intermediary), and white tibialis anterior (wTA, glycolytic). In the fed state, IMCL was significantly elevated in obese compared with lean rats in all three muscle types (soleus: 304%, EDL: 333%, wTA: 394%) in the presence of elevated serum triglycerides but similar levels of free fatty acids (FFA), malonyl-CoA, and total LCACoAs. During starvation, IMCL in soleus remained relatively constant, whereas in both rat groups IMCL increased significantly in wTA and EDL after comparable dynamics of starvation-induced FFA availability. The decreases of malonyl-CoA in wTA and EDL during starvation were more pronounced in lean than in obese rats, although there were no changes in soleus muscles for both groups. The concomitant increase in IMCL with the fall of malonyl-CoA support the concept that, as a reaction to starvation-induced FFA availability, muscle will activate lipid oxidation more the lower its oxidative capacity and then store the rest as IMCL. PMID- 16380390 TI - Endocrine responses to acute and chronic high-altitude exposure (4,300 meters): modulating effects of caloric restriction. AB - High-altitude anorexia leads to a hormonal response pattern modulated by both hypoxia and caloric restriction (CR). The purpose of this study was to compare altitude-induced neuroendocrine changes with or without energy imbalance and to explore how energy sufficiency alters the endocrine acclimatization process. Twenty-six normal-weight, young men were studied for 3 wk. One group [hypocaloric group (HYPO), n = 9] stayed at sea level and consumed 40% fewer calories than required to maintain body weight. Two other groups were deployed to 4,300 meters (Pikes Peak, CO), where one group (ADQ, n = 7) was adequately fed to maintain body weight and the other [deficient group (DEF), n = 10] had calories restricted as above. HYPO experienced a typical CR-induced reduction in many hormones such as insulin, testosterone, and leptin. At altitude, fasting glucose, insulin, and epinephrine exhibited a muted rise in DEF compared with ADQ. Free thyroxine, thyroid-stimulating hormone, and norepinephrine showed similar patterns between the two altitude groups. Morning cortisol initially rose higher in DEF than ADQ at 4,300 meters, but the difference disappeared by day 5. Testosterone increased in both altitude groups acutely but declined over time in DEF only. Adiponectin and leptin did not change significantly from sea level baseline values in either altitude group regardless of energy intake. These data suggest that hypoxia tends to increase blood hormone concentrations, but anorexia suppresses elements of the endocrine response. Such suppression results in the preservation of energy stores but may sacrifice the facilitation of oxygen delivery and the use of oxygen efficient fuels. PMID- 16380391 TI - Cytokine-induced metabolic effects in human adipocytes are independent of endogenous nitric oxide. AB - Nitric oxide (NO) has been recognized as a potential mediator of inflammation induced metabolic alterations, including insulin resistance. However, expression mechanisms and potential roles of endothelial and inducible NO synthases (eNOS and iNOS, respectively) in human adipocytes are poorly understood. In the present study, we aimed to analyze several aspects of NO-related gene expression and metabolite synthesis in basal and inflammation-activated human adipocyte models. eNOS mRNA was highly expressed in omental and to a lesser extent in human subcutaneous adipose tissue biopsies, but not in purified adipocytes, in mesenchymal stem cell (MSC)- and in preadipocyte-derived adipocytes, respectively. Trace amounts of iNOS mRNA were detected in adipose tissue samples of donors with abdominal infection, as opposed to noninfected subjects. Interferon-gamma, in combination with interleukin-1beta or lipopolysaccharide, evoked a transient (4 h < time < 24 h) iNOS mRNA expression in human MSC and preadipocyte-derived adipocytes, respectively. This induction was preceded by cytokine-specific mRNAs. In addition, it was accompanied by an activation of the tetrahydrobiopterin synthesis pathway and by inhibition of peroxisome proliferator-activated receptor-gamma2. In contrast to murine 3T3-L1-derived adipocytes, iNOS protein and NO oxidation products remained undetectable in iNOS mRNA-positive human adipocytes. Accordingly, coadministration of NOS inhibitors (i.e., Nomega-nitro-L-arginine methyl ester, Nomega-monomethyl-L-arginine, and 1400W) had no effects on insulin-mediated glucose uptake and lipolysis. We conclude that, in human adipocytes, endogenous NO is not involved in metabolic regulation during either basal or cytokine-activated conditions. PMID- 16380392 TI - Analysis of glucose metabolism in diabetic rat retinas. AB - This study was conceived in an effort to understand cause and effect relationships between hyperglycemia and diabetic retinopathy. Numerous studies show that hyperglycemia leads to oxidative stress in the diabetic retinas, but the mechanisms that generate oxidative stress have not been resolved. Increased electron pressure on the mitochondrial electron transfer chain, increased generation of cytosolic NADH, and decreases in cellular NADPH have all been cited as possible sources of reactive oxygen species and nitrous oxide. In the present study, excised retinas from control and diabetic rats were exposed to euglycemic and hyperglycemic conditions. Using a microwave irradiation quenching technique to study retinas of diabetic rats in vivo, glucose, glucose-derived metabolites, and NADH oxidation/reduction status were measured. Studying excised retinas in vitro, glycolytic flux, lactate production, and tricarboxylic acid cycle flux were evaluated. Enzymatically assayed glucose 6-phosphate and fructose 6 phosphate were only slightly elevated by hyperglycemia and/or diabetes, but polyols were increased dramatically. Cytosolic NADH-to-NAD ratios were not elevated by hyperglycemia nor by diabetes in vivo or in vitro. Tricarboxylic acid cycle flux was not increased by the diabetic state nor by hyperglycemia. On the other hand, small increases in glycolytic flux were observed with hyperglycemia, but glycolytic flux was always lower in diabetic compared with control animals. An observed decrease in activity of glyceraldehyde-3-phosphate dehydrogenase may be partially responsible for slow glycolytic flux for retinas of diabetic rats. Therefore, it is concluded that glucose metabolism, downstream of hexokinase, is not elevated by hyperglycemia or diabetes. Metabolites upstream of glucose such as the sorbitol pathway (which decreases NADPH) and polyol synthesis are increased. PMID- 16380393 TI - The health at work handbook. PMID- 16380394 TI - Demonstrating the economic value of investments in health at work: not just a measurement problem. PMID- 16380396 TI - Olfactory sensitivity in medical laboratory workers occupationally exposed to organic solvent mixtures. AB - BACKGROUND: Published epidemiological information relating the effects of occupational exposure to organic solvents (OS) to olfaction is limited. AIMS: The objectives of this pilot study were to measure the chemosensory abilities of medical laboratory employees occupationally exposed to OS mixtures, to compare these with control workers employed within the same occupational setting and to correlate chemosensory performance with OS exposure history and with employees' hedonic (pleasantness) perceptions about workplace OS odors. METHODS: Twenty-four medical laboratory employees (OS-exposed technicians plus control workers minimally exposed to OS) completed a health-related questionnaire, a test of pyridine odor detection threshold, along with a gustatory detection threshold test involving aqueous quinine solutions. Estimates of cumulative hours of OS exposure (CSI) were calculated from self-reports. RESULTS: OS-exposed laboratory technicians detected weaker concentrations of pyridine odor. Positive correlations were detected between CSI estimates to both pyridine detection and the degree that participants reported that OS odors were present in the workplace. However, no association was detected between pyridine detection and how unpleasant workplace OS odors were perceived. The OS-exposed participants were able to detect weaker concentrations of quinine. Compared to controls, OS exposed workers complained more of experiencing several symptoms while working, including headaches, nasal irritation and mild cognitive impairment. CONCLUSIONS: The results of this cross-sectional pilot study indicated that, compared to controls, medical laboratory technicians exposed to low-level OS mixtures displayed evidence of elevated olfactory sensitivity (hyperosmia) to pyridine odor. The relation of this study's results to chemical intolerance warrants further investigation. PMID- 16380397 TI - Offering physical activity counselling in occupational health care -- does it reach the target group? AB - BACKGROUND: Employees participating in worksite physical activity (PA) interventions are mostly the physically active and healthy ones. PA counselling may therefore have an important role in reaching 'at-risk' employees. AIMS: To examine (i) how PA counselling offered by occupational health care (OHC) providers reaches the target group of physically inactive employees who have intentions to increase PA and (ii) the relations of 12 selected variables to respondents' willingness to participate in PA counselling. METHODS: Questionnaire survey of employees of client companies of OHC providers. RESULTS: Eight of the 19 OHC providers contacted participated and recruited a total of nine client companies to the study. A questionnaire survey was delivered to all the employees of the companies (n = 1349). Fifty-eight percent of employees (n = 784) responded to the survey and half of them (n = 380) belonged to the target group of being physically inactive and intending to increase PA. Only half of the respondents (n = 201) in the target group were willing to participate in counselling. Respondents in small companies were more interested in counselling than employees in large companies as were white-collar workers compared to blue-collar workers. Earlier PA discussions in OHC and intention to increase leisure-time PA were also positively related to willingness. CONCLUSIONS: A counselling offer attached to a survey did not effectively reach the target group of physically inactive employees who were ready to increase their PA. More individually based approaches such as brief conversations during client contacts are needed in OHC to raise the interest in lifestyle issues. PMID- 16380398 TI - Effectiveness of smoking cessation advice for asbestos workers. AB - AIM: To assess effectiveness of smoking cessation advice to asbestos workers at statutory medical examinations. METHODS: Workers are required to be examined every 2 years while continuing potential exposure to asbestos, including cessation advice if smokers. Records of 170 workers seen between 1986 and 2004 in Kent and Canterbury Hospital were reviewed. Respiratory symptoms, signs, lung function, radiological findings and smoking status were analysed. RESULTS: At initial visit 109 (64%) were smokers. Thirty-four of these attended twice. Despite verbal advice, including emphasis on respiratory abnormalities detected in 62%, only three (9%) had ceased smoking by self-reporting at the second visit. CONCLUSION: Despite appropriate counselling regarding the potential risks of asbestos exposure and smoking, the cessation rate among asbestos workers was disappointingly low. Further measures to encourage asbestos workers to enter smoking cessation programmes should be considered. PMID- 16380399 TI - Delivery of health surveillance for hand-arm vibration in the West Midlands. AB - BACKGROUND: Concerns about provider competence and quality of hand-arm vibrations (HAVs) health surveillance programmes were identified by Health & Safety Executive (HSE) inspectors. AIMS: To evaluate health surveillance programmes and compare them with published HSE guidance. To identify deficiencies and areas for improvement in the health surveillance programmes. METHODS: A proforma was developed for the study and used on a sample of 10 local occupational health providers. RESULTS: All 10 organizations were aware of current HSE guidance for health surveillance for HAVs but only a minority (30%) were following it. Occupational health provider training, written procedures and health surveillance delivery were all identified as areas requiring improvement. CONCLUSIONS: The majority of organizations were not following HSE guidance. Occupational health providers undertaking health surveillance for HAV require specific training. PMID- 16380400 TI - Sensitization to king scallop (Pectin maximus) and queen scallop (Chlamys opercularis) proteins. AB - OBJECTIVE: To report a case of occupational asthma and urticaria due to the queen scallop (Chlamys opercularis) and king scallop (Pectin maximus). BACKGROUND: A 40 year-old female worked in a shellfish-processing plant, handling king and queen scallops for 5 years. At the time of investigation, she described a 2-year history of work-related respiratory symptoms. METHODS: Serial peak expiratory flow rate readings were recorded and an OASYS study completed. A workplace visit was undertaken and specific immunoglobulin (IgE) radioallergosorbent (RAST) testing of scallop extracts was performed. RESULTS: The OASYS study was consistent with occupational asthma. RAST testing demonstrated evidence of specific sensitization (IgE) to queen and king scallop. There was also some cross reactivity observed with other shellfish (prawns and crabs). CONCLUSION: Workers exposed to aerosols from scallop species are at risk of occupational asthma and require effective respiratory health surveillance. PMID- 16380402 TI - Plus ca change, plus c'est la meme chose. PMID- 16380403 TI - Code of practice for food handler activities. PMID- 16380405 TI - Reduced enterobacterial and increased staphylococcal colonization of the infantile bowel: an effect of hygienic lifestyle? AB - The modern Western lifestyle may have altered the composition of the commensal microflora. Here, we investigated the first year's intestinal colonization pattern in 99 vaginally delivered Swedish infants and 17 delivered by cesarean section. Rectal swabs obtained at 3 d of age were cultured for aerobic bacteria and fecal samples obtained at 1, 2, 4, and 8 wk and at 6 and 12 mo of age were cultivated quantitatively for aerobic and anaerobic bacteria. Vaginally delivered infants more often had Escherichia coli compared with cesarean section-delivered infants, whereas the latter more frequently carried other enterobacteria, such as Klebsiella and Enterobacter. Independent of delivery mode, it took 2 mo until most infants were colonized by enterobacteria, traditionally the first colonizers. In contrast, coagulase-negative staphylococci colonized 99% of the infants from d 3 onwards. The poor adaptation of staphylococci to the gut was shown by declining population sizes after some weeks. Dominating anaerobes were initially bifidobacteria and clostridia, whereas Bacteroides initially colonized only 30% of vaginally delivered infants and increased very slowly in prevalence. Bacteroides colonization was delayed up to 1 y in cesarean section-delivered compared with vaginally delivered infants. Our results show that some "traditional" fecal bacteria are acquired late today especially in cesarean section-delivered infants, probably due to limited environmental circulation. In their absence, skin bacteria like staphylococci have become the first gut colonizers. PMID- 16380406 TI - The effect of multifunctionality on the rate of evolution in yeast. AB - Multifunctional genes are expected to evolve at lower rates because mutations in such genes that improve one function might often have deleterious effects on other functions. Here we tested for an association between multifunctionality and evolutionary rates in genes of Saccharomyces cerevisiae, and we find a highly significant negative correlation between the number of biological processes in which a gene is involved in and its rate of evolution. However, the magnitude of this effect is small, and the results do not support the notion that multifunctionality limits a gene's rate of evolution. PMID- 16380407 TI - Uteroplacental insufficiency affects epigenetic determinants of chromatin structure in brains of neonatal and juvenile IUGR rats. AB - Intrauterine growth retardation (IUGR) increases the risk of neuroendocrine reprogramming. In the rat, IUGR leads to persistent changes in cerebral mRNA levels. This suggests lasting alterations in IUGR cerebral transcriptional regulation, which may result from changes in chromatin structure. Candidate nutritional triggers for these changes include altered cerebral zinc and one carbon metabolite levels. We hypothesized that IUGR affects cerebral chromatin structure in neonatal and postnatal rat brains. Rats were rendered IUGR by bilateral uterine artery ligation; controls (Con) underwent sham surgery. At day of life 0 (d0), we measured cerebral DNA methylation, histone acetylation, expression of chromatin-affecting enzymes, and cerebral levels of one-carbon metabolites and zinc. At day of life 21 (d21), we measured cerebral DNA methylation and histone acetylation, as well as the caloric content of Con and IUGR rat breast milk. At d0, IUGR significantly decreased genome-wide and CpG island methylation, as well as increased histone 3 lysine 9 (H3/K9) and histone 3 lysine 14 (H3/K14) acetylation in the hippocampus and periventricular white matter, respectively. IUGR also decreased expression of the chromatin-affecting enzymes DNA methyltransferase 1 (DNMT1), methyl-CpG binding protein 2 (MeCP2), and histone deacetylase (HDAC)1 in association with increased cerebral levels of zinc. In d21 female IUGR rats, cerebral CpG DNA methylation remained lower, whereas H3/K9 and H3/K14 hyperacetylation persisted in hippocampus and white matter, respectively. In d21 male rats, IUGR decreased acetylation of H3/K9 and H3/K14 in these respective regions compared with controls. Despite these differences, caloric, fat, and protein content were similar in breast milk from Con and IUGR dams. We conclude that IUGR results in postnatal changes in cerebral chromatin structure and that these changes are sex specific. PMID- 16380408 TI - Denervation in murine fast-twitch muscle: short-term physiological changes and temporal expression profiling. AB - Denervation deeply affects muscle structure and function, the alterations being different in slow and fast muscles. Because the effects of denervation on fast muscles are still controversial, and high-throughput studies on gene expression in denervated muscles are lacking, we studied gene expression during atrophy progression following denervation in mouse tibialis anterior (TA). The sciatic nerve was cut close to trochanter in adult CD1 mice. One, three, seven, and fourteen days after denervation, animals were killed and TA muscles were dissected out and utilized for physiological experiments and gene expression studies. Target cDNAs from TA muscles were hybridized on a dedicated cDNA microarray of muscle genes. Seventy-one genes were found differentially expressed. Microarray results were validated, and the expression of relevant genes not probed on our array was monitored by real-time quantitative PCR (RQ PCR). Nuclear- and mitochondrial-encoded genes implicated in energy metabolism were consistently downregulated. Among genes implicated in muscle contraction (myofibrillar and sarcoplasmic reticulum), genes typical of fast fibers were downregulated, whereas those typical of slow fibers were upregulated. Electrophoresis and Western blot showed less pronounced changes in myofibrillar protein expression, partially confirming changes in gene expression. Isometric tension of skinned fibers was little affected by denervation, whereas calcium sensitivity decreased. Functional studies in mouse extensor digitorum longus muscle showed prolongation in twitch time parameters and shift to the left in force-frequency curves after denervation. We conclude that, if studied at the mRNA level, fast muscles appear not less responsive than slow muscles to the interruption of neural stimulation. PMID- 16380409 TI - Induction chemotherapy for larynx preservation: patient selection or therapeutic effect? PMID- 16380410 TI - Extranodal natural killer T-cell lymphoma, nasal-type: a prognostic model from a retrospective multicenter study. AB - PURPOSE: Patients with natural killer T (NK/T) -cell lymphomas have poor survival outcome, and for this condition there is no optimal therapy. The purpose of this study was to design a prognostic model specifically for extranodal NK/T-cell lymphoma, which can identify high-risk patients who need more aggressive therapy. PATIENTS AND METHODS: This multicenter retrospective study was comprised of 262 patients who were diagnosed with NK/T-cell lymphoma. RESULTS: After a median follow-up duration of 51.2 months, 5-year overall survival rate in 262 patients was 49.5%. Prognostic factors for survival were "B" symptoms (P = .0003; relative risk, 2.202; 95% CI, 1.446 to 3.353), stage (P = .0006; relative risk, 2.366; 95% CI, 1.462 to 3.828), lactate dehydrogenase (LDH) level (P = .0005; relative risk, 2.278; 95% CI, 1.442 to 3.598), and regional lymph nodes (P = .0044; relative risk, 1.546; 95% CI, 1.009 to 2.367). Of 262 patients, 219 had complete information on four parameters. We identified four different risk groups: group 1, no adverse factor; group 2, one factor; group 3, two factors; and group 4, three or four factors. The new model showed a superior prognostic discrimination as compared with the International Prognostic Index (IPI). Notably, the distribution of patients was balanced when a new model was adopted (group 1, 27%; group 2, 31%; group 3, 20%; group 4, 22%), whereas 81% of patients were categorized as low or low-intermediate risks using IPI. CONCLUSION: The newly proposed model for extranodal NK/T-cell lymphoma demonstrated a more balanced distribution of patients into four groups with better prognostic discrimination as compared with the IPI. PMID- 16380411 TI - Outcome of critically ill allogeneic hematopoietic stem-cell transplantation recipients: a reappraisal of indications for organ failure supports. AB - PURPOSE: Because the overall outcome of critically ill hematologic patients has improved, we evaluated the short-term and long-term outcomes of the poor risk subgroup of allogeneic hematopoietic stem-cell transplantation (HSCT) recipients requiring admission to the intensive care unit (ICU). PATIENTS AND METHODS: This was a retrospective multicenter study of allogeneic HSCT recipients admitted to the ICU between 1997 and 2003. RESULTS: Two hundred nine critically ill allogeneic HSCT recipients were included in the study. Admission in the ICU occurred during the engraftment period (< or = 30 days after transplantation) for 70 of the patients and after the engraftment period for 139 patients. The overall in-ICU, in-hospital, 6-month, and 1-year survival rates were 48.3%, 32.5%, 27.2%, and 21%, respectively. Mechanical ventilation was required in 122 patients and led to a dramatic decrease in survival rates, resulting in in-ICU, in-hospital, 6 month, and 1-year survival rates of 18%, 15.6%, 14%, and 10.6%, respectively. Mechanical ventilation, elevated bilirubin level, and corticosteroid treatment for the indication of active graft-versus-host disease (GVHD) were independent predictors of death in the whole cohort. In the subgroup of patients requiring mechanical ventilation, associated organ failures, such as shock and liver dysfunction, were independent predictors of death. ICU admission during engraftment period was associated with acceptable outcome in mechanically ventilated patients, whereas patients with late complications of HSCT in the setting of active GVHD had a poor outcome. CONCLUSION: Extensive unlimited intensive care support is justified for allogeneic HSCT recipients with complications occurring during the engraftment period. Conversely, initiation or maintenance of mechanical ventilation is questionable in the setting of active GVHD. PMID- 16380412 TI - Antibody pretargeting advances cancer radioimmunodetection and radioimmunotherapy. AB - This article reviews the methods of pretargeting, which involve separating the targeting antibody from the subsequent delivery of an imaging or therapeutic agent that binds to the tumor-localized antibody. This provides enhanced tumor:background ratios and the delivery of a higher therapeutic dose than when antibodies are directly conjugated with radionuclides, as currently practiced in cancer radioimmunotherapy. We describe initial promising clinical results using streptavidin-antibody constructs with biotin-radionuclide conjugates in the treatment of patients with malignant gliomas, and of bispecific antibodies with hapten-radionuclides in the therapy of tumors expressing carcinoembryonic antigen, such as medullary thyroid and small-cell lung cancers. PMID- 16380413 TI - Preoperative serum tissue factor levels are an independent prognostic factor in patients with ovarian carcinoma. AB - PURPOSE: Tissue factor (TF) is a procoagulant that plays an important part in tumor angiogenesis. We sought to determine the role of preoperative serum TF levels in predicting clinical outcome in patients with ovarian cancer. MATERIALS AND METHODS: TF expression was determined by reverse transcriptase polymerase chain reaction in ovarian cell lines. Using enzyme-linked immunosorbent assay, we assessed preoperative serum TF levels in 98 women with invasive epithelial ovarian carcinoma, 30 with low malignant potential (LMP) tumors, 16 with benign tumors, and a separate validation group of 39 women with adnexal masses. Clinical information was gathered from chart review. RESULTS: TF was expressed in four of the five ovarian cancer cell lines, but absent in the nontransformed cells. Ovarian cancer patients had a median preoperative serum TF level of 85.2 pg/mL, which was significantly higher than in those with LMP tumors (12.8 pg/mL; P < .01) and benign adnexal disease (30.7 pg/mL; P < .01). TF >or= 190 pg/mL was significantly associated with decreased patient survival (P < .01). After adjusting for other clinical variables in a multivariate Cox regression model, TF >or= 190 pg/mL was an independent prognostic factor (P < .01). Analysis of serum TF levels from the validation set confirmed that high TF (>or=190 pg/mL) was associated with a 3.4-fold increase in risk of death from disease (P = .02) and shorter survival (P = .01). CONCLUSION: Preoperative serum TF levels are significantly elevated in patients with ovarian carcinoma. Elevated preoperative TF level is an independent prognostic factor for death from disease. PMID- 16380414 TI - DeltaTAp73 upregulation correlates with poor prognosis in human tumors: putative in vivo network involving p73 isoforms, p53, and E2F-1. AB - PURPOSE: Although full-length TAp73 variants largely mimic p53 suppressor activities, the transactivation-deficient transcripts DeltaTAp73 exert an oncogenic effect by inactivating p53 and TAp73 suppressor properties. Additionally, DeltaTAp73 may cooperate with oncogenic RAS to induce cell transformation, confer drug resistance, and induce the phosphorylation of phosphorylated Rb. Here, we study the expression of TAp73 and DeltaTAp73 variants and assess possible associations with E2F-1, p53 and K-ras status. We address the possible clinical relevance of alterations in these genes. PATIENTS AND METHODS: We determine in 113 colon and 60 breast cancer patients (a) the expression levels of TAp73, DeltaTAp73 (DeltaEx2p73, DeltaEx2/3p73, and DeltaNp73), and E2F-1 transcripts by quantitative real-time reverse transcriptase polymerase chain reaction (PCR); (b) mutations in the first exon of K-ras by PCR-single-stranded confirmational polymorphism; and (c) p53 status by immunohistochemistry. Tumor characteristics were examined in each patient. RESULTS: Both suppressor and oncogenic isoforms of TP73 were significantly coupregulated in tumor tissues. Associations were observed between (a) p53 wild type status and upregulation of some TP73 variants; (b) overexpression of E2F-1 and some TP73 forms; and (c) upregulation of DeltaTAp73 variants and advanced pathologic stage, lymph node metastasis, vascular invasion, presence of polyps, and tumor localization. CONCLUSION: Overexpression of TP73 variants in tumor tissues indicates that they may be involved in colon and breast carcinogenesis. The association between upregulation of DeltaTAp73 isoforms and poor prognosis features, specifically advanced tumor stage, suggests that they may be of practical clinical prognostic value. Interestingly, the in vivo associations identified here may indicate a functional network involving p73 variants, p53, and E2F-1. PMID- 16380415 TI - Single-cycle induction chemotherapy selects patients with advanced laryngeal cancer for combined chemoradiation: a new treatment paradigm. AB - PURPOSE: Primary chemoradiotherapy in patients with advanced laryngeal cancer can achieve high rates of organ preservation without sacrificing survival compared with radiation alone or conventional laryngectomy. Appropriate selection of patients for organ preservation approaches could enhance overall treatment outcome and quality of life. We conducted a phase II organ preservation trial for patients with stage III and IV larynx cancer to determine whether late salvage surgery rates could be decreased and survival improved by selecting patients for organ preservation based on response to a single cycle of induction chemotherapy. PATIENTS AND METHODS: The chemotherapy was cisplatin 100 mg/m2 on day 1 and fluorouracil 1,000 mg/m(2)/d for 5 days. Patients who achieved less than 50% response had immediate laryngectomy. Patients who achieved more than 50% response went on to concurrent chemoradiotherapy. Histologic complete responders after chemoradiotherapy received two more cycles of chemotherapy. Patients with residual disease after chemoradiotherapy had planned salvage surgery. RESULTS: Of 97 eligible patients, 73 (75%) achieved more than 50% response and received chemoradiotherapy. A total of 29 patients (30%) had salvage surgery; 19 patients (20%) had early salvage surgery after the single cycle of induction chemotherapy, three patients (3%) had late salvage surgery after chemoradiotherapy, six patients (6%) eventually had salvage surgery for recurrence, and one patient had laryngectomy for chondroradionecrosis. The median follow-up time was 41.9 months. The overall survival rate at 3 years is 85%. The cause-specific survival rate was 87%. Larynx preservation was achieved in 69 patients (70%). CONCLUSION: These results confirm excellent larynx preservation and improved overall survival rates compared with historical results. PMID- 16380416 TI - Mesozoic aviary takes form. PMID- 16380417 TI - Cinnamoyl-CoA reductase, a key enzyme in lignin biosynthesis, is an effector of small GTPase Rac in defense signaling in rice. AB - OsRac1, one of the Rac/Rop family of small GTPases, plays important roles in defense responses, including a role in the production of reactive oxygen species mediated by NADPH oxidase. We have identified an effector of OsRac1, namely rice (Oryza sativa) cinnamoyl-CoA reductase 1 (OsCCR1), an enzyme involved in lignin biosynthesis. Lignin, which is polymerized through peroxidase activity by using H(2)O(2) in the cell wall, is an important factor in plant defense responses, because it presents an undegradable mechanical barrier to most pathogens. Expression of OsCCR1 was induced by a sphingolipid elicitor, suggesting that OsCCR1 participates in defense signaling. In in vitro interaction and two-hybrid experiments, OsRac1 was shown to bind OsCCR1 in a GTP-dependent manner. Moreover, the interaction of OsCCR1 with OsRac1 led to the enzymatic activation of OsCCR1 in vitro. Transgenic cell cultures expressing the constitutively active OsRac1 accumulated lignin through enhanced CCR activity and increased reactive oxygen species production. Thus, it is likely that OsRac1 controls lignin synthesis through regulation of both NADPH oxidase and OsCCR1 activities during defense responses in rice. PMID- 16380418 TI - Atherosclerosis quantitative trait loci are sex- and lineage-dependent in an intercross of C57BL/6 and FVB/N low-density lipoprotein receptor-/- mice. AB - Atherosclerosis is a complex disease that is affected by environmental as well as genetic factors. The aim of the present study was to identify loci of atherosclerosis susceptibility in a cross of atherosclerosis-susceptible C57BL/6 and atherosclerosis-resistant FVB/N mice on the low-density lipoprotein (LDL) receptor (LDLR)-deficient background (LDLR(-/-)) and to test whether these loci are affected by lineage. A total of 459 F(2)s were generated in two ways: In cross "mB6xfFVB," male B6.LDLR(-/-) mice were crossed to female FVB.LDLR(-/-) mice to generate 107 female and 112 male F(2)s. In cross "mFVBxfB6," male FVB.LDLR(-/-) mice were crossed to female B6.LDLR(-/-) mice to generate 120 female and 120 male F(2)s. Animals were phenotyped for cross-sectional atherosclerotic lesion area at the aortic root, and a genome scan was carried out with 192 microsatellite markers. Quantitative trait locus mapping revealed significant loci of atherosclerosis susceptibility on chromosomes 3, 10, and 12. On chromosome 10 maximal logarithm of odds (LOD) scores of 13.1 (D10Mit16, 16 cM) and 5.7 (D10Mit168, 9 cM) were found in female and male mice, respectively. On chromosome 3, a maximal LOD score of 5.1 (D3Mit45, 79 cM) was detected only in females. On proximal chromosome 12 significant LOD scores were lineage-dependent, with maximal LOD scores of 3.9 (D12Mit82, 3 cM) and 4.8 (D12Mit189, 24 cM) present only in female mice of cross mB6xfFVB and male mice of cross mFVBxfB6, respectively. We conclude that, in this intercross, loci of atherosclerosis susceptibility are in part sex- and lineage-dependent. Awareness of these complexities may have major consequences for the identification of atherosclerosis susceptibility genes by quantitative trait locus mapping. PMID- 16380419 TI - Profile of Xiaodong Wang. PMID- 16380420 TI - More barrels from the viral tree of life. PMID- 16380421 TI - Specific, efficient, and selective inhibition of prokaryotic translation initiation by a novel peptide antibiotic. AB - Many known antibiotics target the translational apparatus, but none of them can selectively inhibit initiation of protein synthesis and/or is prokaryotic specific. This article describes the properties of GE81112, an effective and prokaryotic-specific initiation inhibitor. GE81112 is a natural tetrapeptide produced by a Streptomyces sp. identified by an in vitro high-throughput screening test developed to find inhibitors of the prokaryotic translational apparatus preferentially acting on steps other than elongation. In vivo GE81112 inhibits protein synthesis but not other cell functions such as DNA duplication, transcription, and cell wall synthesis. In vitro GE81112 was found to target the 30S ribosomal subunit and to interfere with both coded and noncoded P-site binding of fMet-tRNA, thereby selectively inhibiting formation of the 30S initiation complex. PMID- 16380422 TI - Crystal structure of phycocyanobilin:ferredoxin oxidoreductase in complex with biliverdin IXalpha, a key enzyme in the biosynthesis of phycocyanobilin. AB - Phytobilins (light harvesting and photoreceptor pigments in higher plants, algae, and cyanobacteria) are synthesized from biliverdin IXalpha (BV) by ferredoxin dependent bilin reductases (FDBRs). Phycocyanobilin:ferredoxin oxidoreductase (PcyA), one such FDBR, is a new class of radical enzymes that require neither cofactors nor metals and serially reduces the vinyl group of the D-ring and A ring of BV using four electrons from ferredoxin to produce phycocyanobilin, one of the phytobilins. We have determined the crystal structure of PcyA from Synechocystis sp. PCC 6803 in complex with BV, revealing the first tertiary structure of an FDBR family member. PcyA is folded in a three-layer alpha/beta/alpha sandwich structure, in which BV in a cyclic conformation is positioned between the beta-sheet and C-terminal alpha-helices. The basic patch on the PcyA surface near the BV molecule may provide a binding site for acidic ferredoxin, allowing direct transfer of electrons to BV. The orientation of BV is definitely fixed in PcyA by several hydrophilic interactions and the shape of the BV binding pocket of PcyA. We propose the mechanism by which the sequential reduction of the D- and A-rings is controlled, where Asp-105, located between the two reduction sites, would play the central role by changing its conformation during the reaction. Homology modeling of other FDBRs based on the PcyA structure fits well with previous genetic and biochemical data, thereby providing a structural basis for the reaction mechanism of FDBRs. PMID- 16380423 TI - Modeling the spatiotemporal cortical activity associated with the line-motion illusion in primary visual cortex. AB - Our large-scale computational model of the primary visual cortex that incorporates orientation-specific, long-range couplings with slow NMDA conductances operates in a fluctuating dynamic state of intermittent desuppression (IDS), which captures the behavior of coherent spontaneous cortical activity, as revealed by in vivo optical imaging based on voltage-sensitive dyes. Here, we address the functional significance of the IDS cortical operating points by investigating our model cortex response to the Hikosaka line-motion illusion (LMI) stimulus-a cue of a quickly flashed stationary square followed a few milliseconds later by a stationary bar. As revealed by voltage-sensitive dye imaging, there is an intriguing similarity between the cortical spatiotemporal activity in response to (i) the Hikosaka LMI stimulus and (ii) a small moving square. This similarity is believed to be associated with the preattentive illusory motion perception. Our numerical cortex produces similar spatiotemporal patterns in response to the two stimuli above, which are both in very good agreement with experimental results. The essential network mechanisms underpinning the LMI phenomenon in our model are (i) the spatiotemporal structure of the LMI input as sculpted by the lateral geniculate nucleus, (ii) a priming effect of the long-range NMDA-type cortical coupling, and (iii) the NMDA conductance-voltage correlation manifested in the IDS state. This mechanism in our model cortex, in turn, suggests a physiological underpinning for the LMI associated patterns in the visual cortex of anaesthetized cat. PMID- 16380424 TI - Morphological affinities of the Sahelanthropus tchadensis (Late Miocene hominid from Chad) cranium. AB - The recent reconstruction of the Sahelanthropus tchadensis cranium (TM 266-01-60 1) provides an opportunity to examine in detail differences in cranial shape between this earliest-known hominid, African apes, and other hominid taxa. Here we compare the reconstruction of TM 266-01-60-1 with crania of African apes, humans, and several Pliocene hominids. The results not only confirm that TM 266 01-60-1 is a hominid but also reveal a unique mosaic of characters. The TM 266-01 60-1 reconstruction shares many primitive features with chimpanzees but overall is most similar to Australopithecus, particularly in the basicranium. However, TM 266-01-60-1 is distinctive in having the combination of a short subnasal region associated with a vertical upper face that projects substantially in front of the neurocranium. Further research is needed to determine the evolutionary relationships between Sahelanthropus and the known Miocene and Pliocene hominids. PMID- 16380425 TI - Experimental support for the "E pathway hypothesis" of coupled transmembrane e- and H+ transfer in dihemic quinol:fumarate reductase. AB - Reconciliation of apparently contradictory experimental results obtained on the quinol:fumarate reductase, a diheme-containing respiratory membrane protein complex from Wolinella succinogenes, was previously obtained by the proposal of the so-called "E pathway hypothesis." According to this hypothesis, transmembrane electron transfer via the heme groups is strictly coupled to cotransfer of protons via a transiently established pathway thought to contain the side chain of residue Glu-C180 as the most prominent component. Here we demonstrate that, after replacement of Glu-C180 with Gln or Ile by site-directed mutagenesis, the resulting mutants are unable to grow on fumarate, and the membrane-bound variant enzymes lack quinol oxidation activity. Upon solubilization, however, the purified enzymes display approximately 1/10 of the specific quinol oxidation activity of the wild-type enzyme and unchanged quinol Michaelis constants, K(m). The refined x-ray crystal structures at 2.19 A and 2.76 A resolution, respectively, rule out major structural changes to account for these experimental observations. Changes in the oxidation-reduction heme midpoint potential allow the conclusion that deprotonation of Glu-C180 in the wild-type enzyme facilitates the reoxidation of the reduced high-potential heme. Comparison of solvent isotope effects indicates that a rate-limiting proton transfer step in the wild-type enzyme is lost in the Glu-C180 --> Gln variant. The results provide experimental evidence for the validity of the E pathway hypothesis and for a crucial functional role of Glu-C180. PMID- 16380426 TI - Enhancer-independent Mu transposition from two topologically distinct synapses. AB - Transposition of Mu is strictly dependent on a specific orientation of the left (L) and right (R) ends of Mu and a distant enhancer site (E) located on supercoiled DNA. Five DNA crossings are trapped in the three-site synapse, two of which are contributed by the interwrapping of L and R. To determine the contribution of E to the interwrapping of Mu ends, we examined the topology of the LR synapse under two different enhancer-independent reaction conditions. One of these conditions, which also alleviates the requirement for a specific orientation of Mu ends, revealed two topologically distinct arrangements of the ends. In their normal relative orientation, L and R were either plectonemically interwrapped or aligned by random collision. Addition of the enhancer to this system channeled synapsis toward the interwrapped pathway. When the ends were in the wrong relative orientation, synapsis occurred exclusively by random collision. In the second enhancer-independent condition, which retains the requirement for a specific orientation of Mu ends, synapsis of L and R was entirely by interwrapping. The two distinct kinds of synapses also were identified by gel electrophoresis. We discuss these results in the context of the "topological filter" model and consider the many contributions the enhancer makes to the biologically relevant interwrapped synapse. PMID- 16380427 TI - The evolutionary history of Cys-tRNACys formation. AB - The recent discovery of an alternate pathway for indirectly charging tRNA(Cys) has stimulated a re-examination of the evolutionary history of Cys-tRNA(Cys) formation. In the first step of the pathway, O-phosphoseryl-tRNA synthetase charges tRNA(Cys) with O-phosphoserine (Sep), a precursor of the cognate amino acid. In the following step, Sep-tRNA:Cys-tRNA synthase (SepCysS) converts Sep to Cys in a tRNA-dependent reaction. The existence of such a pathway raises several evolutionary questions, including whether the indirect pathway is a recent evolutionary invention, as might be implied from its localization to the Euryarchaea, or, as evidence presented here indicates, whether this pathway is more ancient, perhaps already in existence at the time of the last universal common ancestral state. A comparative phylogenetic approach is used, combining evolutionary information from protein sequences and structures, that takes both the signature of horizontal gene transfer and the recurrence of the full canonical phylogenetic pattern into account, to document the complete evolutionary history of cysteine coding and understand the nature of this process in the last universal common ancestral state. Resulting from the historical study of tRNA(Cys) aminoacylation and the integrative perspective of sequence, structure, and function are 3D models of O-phosphoseryl-tRNA synthetase and SepCysS, which provide experimentally testable predictions regarding the identity and function of key active-site residues in these proteins. The model of SepCysS is used to suggest a sulfhydrylation reaction mechanism, which is predicted to occur at the interface of a SepCysS dimer. PMID- 16380428 TI - Increase in TNF-alpha and inducible nitric oxide synthase-expressing dendritic cells in psoriasis and reduction with efalizumab (anti-CD11a). AB - We find that CD11c(+) cells with many markers of dendritic cells (DCs) are a major cell type in the skin lesions of psoriasis. These CD11c(+) cells, which are evident in both epidermis and dermis, are the sites for the expression of two mediators of inflammation, inducible nitric oxide synthase (iNOS) and TNF-alpha in diseased skin. These cells express HLA-DR, CD40, and CD86, lack the Langerin and CD14 markers of Langerhans cells and monocytes, respectively, and to a significant extent express the DC maturation markers DC-LAMP and CD83. Treatment of psoriasis with efalizumab (anti-CD11a, Raptiva) strongly reduces infiltration by these DCs in patients responding to this agent. Disease activity after therapy was more related to DC infiltrates and iNOS mRNA levels than T cell infiltrates, and CD11c(+) cells responded more quickly to therapy than epidermal keratinocytes. Our results suggest that a type of DC, which resembles murine "Tip DCs" that can accumulate during infection, has proinflammatory effects in psoriasis through nitric oxide and TNF-alpha production, and can be an important target for suppressive therapies. PMID- 16380429 TI - Simultaneous dopamine and single-unit recordings reveal accumbens GABAergic responses: implications for intracranial self-stimulation. AB - Intracranial self-stimulation (ICS) is a motivated behavior that results from contingent activation of the brain reward system. ICS with stimulating electrodes placed in the medial forebrain bundle (MFB) is particularly robust. However, the neurons that course through this pathway use a variety of neurotransmitters including dopamine and GABA. For this reason, the neurotransmitters that are central to this behavior, and the specific roles that they subserve, remain unclear. Here, we used extracellular electrophysiology and cyclic voltammetry at the same electrode in awake rats to simultaneously examine cell firing and dopamine release in the nucleus accumbens (NAc) during ICS and noncontingent stimulation of the MFB. ICS elicited dopamine release in the NAc and produced coincident time-locked changes (predominantly inhibitions) in the activity of a subset of NAc neurons. Similar responses were elicited with noncontingent stimulations. The changes in firing rate induced by noncontingent stimulations were reversed by the GABA(A) receptor antagonist bicuculline. Most time-locked unit activity was unaffected by D1 or D2-like dopamine-receptor antagonists, or by inhibition of evoked dopamine release, although, for a minority of units, the D1 dopamine-receptor antagonist SCH23390 attenuated neural activity. Thus, neurons in the NAc are preferentially inhibited by GABA(A) receptors after MFB stimulation, a mechanism that may also be important in ICS. PMID- 16380430 TI - Asymmetry in corticofugal modulation of frequency-tuning in mustached bat auditory system. AB - Focal electric stimulation of the auditory cortex is well suited for exploration of the function of the corticofugal (descending) system and the neural mechanism of plasticity in the central auditory system, because it evokes changes in frequency-tuning, called best frequency (BF) shifts, as does auditory fear conditioning. The Doppler-shifted constant frequency (DSCF) area of the primary auditory cortex of the mustached bat is highly specialized for fine frequency analysis. Focal electric stimulation of the DSCF area evokes the BF shifts of ipsilateral cortical and collicular neurons away from the BF of stimulated neurons, whereas the stimulation evokes the BF shifts of contralateral cortical and collicular neurons either toward or away from the stimulated BF. The direction of contralateral BF shifts shows a flip-flop, depending on the spatial relationship between the stimulated and recorded neurons. This asymmetry in corticofugal modulation is mostly, if not totally, created by two subdivisions of the stimulated DSCF area that transmit signals to the contralateral DSCF area, presumably through the corpus callosum. This intriguing asymmetry in corticofugal modulation presumably functions for equalization of the reorganization of the frequency maps of the DSCF areas and subcortical auditory nuclei on both sides. PMID- 16380431 TI - CREB-binding protein controls response to cocaine by acetylating histones at the fosB promoter in the mouse striatum. AB - Remodeling chromatin is essential for cAMP-regulated gene expression, necessary not only for development but also for memory storage and other enduring mental states. Histone acetylation and deacetylation mediate long-lasting forms of synaptic plasticity in Aplysia as well as cognition in mice. Here, we show that histone acetylation by the cAMP-response element binding protein (CREB)-binding protein (CBP) mediates sensitivity to cocaine by regulating expression of the fosB gene and its splice variant, DeltafosB, a transcription factor previously implicated in addiction. Using the chromatin immunoprecipitation assay with antibodies against histone H4 or CBP, we find that CBP is recruited to the fosB promoter to acetylate histone H4 in response to acute exposure to cocaine. We show that mutant mice that lack one allele of the CBP gene and have normal levels of fosB expression are less sensitive to chronic (10-day) administration of cocaine than are wild-type mice. This decreased sensitivity is correlated with decreased histone acetylation and results in decreased fosB expression and diminished accumulation of DeltafosB. Thus, CBP, which forms part of the promoter complex with CREB, mediates sensitivity to cocaine by acetylating histones. PMID- 16380432 TI - Involvement of KU80 in T-DNA integration in plant cells. AB - In Agrobacterium-mediated genetic transformation of plant cells, the bacterium exports a well defined transferred DNA (T-DNA) fragment and a series of virulence proteins into the host cell. Following its nuclear import, the single-stranded T DNA is stripped of its escorting proteins, most likely converts to a double stranded (ds) form, and integrates into the host genome. Little is known about the precise mechanism of T-DNA integration in plants, and no plant proteins specifically associated to T-DNA have been identified. Here we report the direct involvement of KU80, a protein that binds dsT-DNA intermediates. We show that ku80-mutant Arabidopsis plants are defective in T-DNA integration in somatic cells, whereas KU80-overexpressing plants exhibit increased susceptibility to Agrobacterium infection and increased resistance to DNA-damaging agents. The direct interaction between dsT-DNA molecules and KU80 in planta was confirmed by immunoprecipitation of KU80 dsT-DNA complexes from Agrobacterium-infected plants. Transformation of KU80-overexpressing plants with two separate T-DNA molecules resulted in an increased rate of extrachromosomal T-DNA to T-DNA recombination, indicating that KU80 bridges between dsT-DNAs and double-strand breaks. This last result further supports the notion that integration of T-DNA molecules occurs through ds intermediates and requires active participation of the host's nonhomologous end-joining repair machinery. PMID- 16380434 TI - Acute severe myopathy following a single infusion of omeprazole. PMID- 16380433 TI - Identification of BSPRY as a novel auxiliary protein inhibiting TRPV5 activity. AB - Transient receptor potential vallinoid 5 (TRPV5) and TRPV6 are the most Ca2+ selective members of the TRP superfamily and are essential for active Ca2+ (re)absorption in epithelia. However, little is known about intracellular proteins that regulate the activity of these channels. This study identified BSPRY (B-box and SPRY-domain containing protein) as a novel factor involved in the control of TRPV5. The interaction between BSPRY and TRPV5 by GST pull-down and co-immunoprecipitation assays was demonstrated. BSPRY showed co-localization with TRPV5 in mouse kidney. Expression of BSPRY resulted in a significant reduction of the Ca2+ influx in Madin-Darby Canine Kidney cells that stably express TRPV5 without affecting channel cell-surface abundance. Finally, BSPRY expression in kidney was increased in 25-hydroxyvitamin D3-1alpha-hydroxylase knockout mice, suggesting an inverse regulation by vitamin D3. Together, these results demonstrate the physiologic role of the novel protein BSPRY in the regulation of epithelial Ca2+ transport via negative modulation of TRPV5 activity. PMID- 16380435 TI - Catatonic schizophrenia and the use of memantine. AB - OBJECTIVE: To report a case of catatonic schizophrenia treated with memantine that resulted in a rapid reduction of catatonic symptoms. CASE SUMMARY: A 68-year old male with catatonic schizophrenia presented with bizarre and catatonic behavior manifested by mutism, waxy flexibility, immobility, staring, nonresponsiveness to verbal commands, grimacing, rigidity, and posturing. During the course of his hospitalization, he was treated with memantine up to 10 mg/day. The catatonia responded rapidly and significantly to memantine. DISCUSSION: Few treatment options are available for the management of catatonia. Lorazepam is generally the agent that has been used most often and has had varying degrees of success. Our report adds to a recently published case that also suggested that memantine might be useful for the management of catatonic behaviors in a patient with schizophrenia. While its mechanism of action is unknown, it has been thought that glutamate antagonists, such as memantine, may be beneficial in catatonic schizophrenia due to a glutamatergic dysfunction present in catatonic patients. It also has been hypothesized that there is a decrease in gamma-aminobutyric acid released to the supplementary motor areas, resulting in less glutamate inhibition. This results in a net effect of glutamatergic hyperfunction in the striatum, which may produce catatonia. Based on this hypothesis, memantine would be beneficial as an N-methyl-D-aspartic acid antagonist to decrease the amount of glutamate in the striatum and thereby relieve symptoms of catatonia. However, these potential benefits must be weighed against recently reported adverse reactions associated with memantine use, namely, psychosis and seizures. CONCLUSIONS: Memantine produced a rapid and significant reduction in catatonic symptoms in our patient with catatonic schizophrenia. PMID- 16380436 TI - Analysis of severe adverse events related to the use of mifepristone as an abortifacient. AB - BACKGROUND: The systematic analysis of morbidity and mortality for the Food and Drug Administration (FDA)-approved medical abortion regimen using mifepristone is possible using data from the FDA's Adverse Event Reporting System. OBJECTIVE: To assess mifepristone's mortality, morbidity, sentinel events, and quality of postmarketing surveillance using mifepristone adverse event reports (AERs). METHODS: Six hundred seven unique mifepristone AERs submitted to the FDA over a 4 year span were coded using the National Cancer Institute's Common Terminology Criteria for Adverse Events. Coding was based only on data in AERs and may underestimate severity and treatment rendered. Two board-certified obstetrician/gynecologists, the authors, made individual evaluations, compared them, and agreed upon final coding. RESULTS: The most frequent AERs were hemorrhage (n = 237) and infection (66). Hemorrhages included 1 fatal, 42 life threatening, and 168 serious cases; 68 required transfusions. Infections included 7 cases of septic shock (3 fatal, 4 life threatening) and 43 cases requiring parenteral antibiotics. Surgical interventions were required in 513 cases (235 emergent, 278 nonemergent). Emergent cases included 17 ectopic pregnancies (11 ruptured). Second trimester viability was documented in 22 cases (9 lost to follow-up, 13 documented fetal outcome). Of the 13 documented cases, 9 were terminated without comment on fetal morphology, 1 was enrolled in fetal registry, and 3 fetuses were diagnosed with serious malformations, suggesting a malformation rate of 23%. CONCLUSIONS: Hemorrhage and infection are the leading causes of mifepristone-related morbidity and mortality. AERs relied upon by the FDA to monitor mifepristone's postmarketing safety are grossly deficient due to extremely poor quality. PMID- 16380437 TI - p53 functions as a negative regulator of osteoblastogenesis, osteoblast-dependent osteoclastogenesis, and bone remodeling. AB - p53 is a well known tumor suppressor. We show that p53 also regulates osteoblast differentiation, bone formation, and osteoblast-dependent osteoclast differentiation. Indeed, p53(-/-) mice display a high bone mass phenotype, and p53(-/-) osteoblasts show accelerated differentiation, secondary to an increase in expression of the osteoblast differentiation factor osterix, as a result. Reporter assays indicate that p53 represses osterix transcription by the minimal promoter in a DNA-binding-independent manner. In addition, p53(-/-) osteoblasts have an enhanced ability to favor osteoclast differentiation, in association with an increase in expression of macrophage-colony stimulating factor, which is under the control of osterix. Furthermore, inactivating p53 is sufficient to rescue the osteoblast differentiation defects observed in mice lacking c-Abl, a p53 interacting protein. Thus, these results identify p53 as a novel regulator of osteoblast differentiation, osteoblast-dependent osteoclastogenesis, and bone remodeling. PMID- 16380438 TI - Pax3 and Pax7 have distinct and overlapping functions in adult muscle progenitor cells. AB - The growth and repair of skeletal muscle after birth depends on satellite cells that are characterized by the expression of Pax7. We show that Pax3, the paralogue of Pax7, is also present in both quiescent and activated satellite cells in many skeletal muscles. Dominant-negative forms of both Pax3 and -7 repress MyoD, but do not interfere with the expression of the other myogenic determination factor, Myf5, which, together with Pax3/7, regulates the myogenic differentiation of these cells. In Pax7 mutants, satellite cells are progressively lost in both Pax3-expressing and -nonexpressing muscles. We show that this is caused by satellite cell death, with effects on the cell cycle. Manipulation of the dominant-negative forms of these factors in satellite cell cultures demonstrates that Pax3 cannot replace the antiapoptotic function of Pax7. These findings underline the importance of cell survival in controlling the stem cell populations of adult tissues and demonstrate a role for upstream factors in this context. PMID- 16380439 TI - Coupling of the nucleus and cytoplasm: role of the LINC complex. AB - The nuclear envelope defines the barrier between the nucleus and cytoplasm and features inner and outer membranes separated by a perinuclear space (PNS). The inner nuclear membrane contains specific integral proteins that include Sun1 and Sun2. Although the outer nuclear membrane (ONM) is continuous with the endoplasmic reticulum, it is nevertheless enriched in several integral membrane proteins, including nesprin 2 Giant (nesp2G), an 800-kD protein featuring an NH(2)-terminal actin-binding domain. A recent study (Padmakumar, V.C., T. Libotte, W. Lu, H. Zaim, S. Abraham, A.A. Noegel, J. Gotzmann, R. Foisner, and I. Karakesisoglou. 2005. J. Cell Sci. 118:3419-3430) has shown that localization of nesp2G to the ONM is dependent upon an interaction with Sun1. In this study, we confirm and extend these results by demonstrating that both Sun1 and Sun2 contribute to nesp2G localization. Codepletion of both of these proteins in HeLa cells leads to the loss of ONM-associated nesp2G, as does overexpression of the Sun1 lumenal domain. Both treatments result in the expansion of the PNS. These data, together with those of Padmakumar et al. (2005), support a model in which Sun proteins tether nesprins in the ONM via interactions spanning the PNS. In this way, Sun proteins and nesprins form a complex that links the nucleoskeleton and cytoskeleton (the LINC complex). PMID- 16380440 TI - Nuclear congression is driven by cytoplasmic microtubule plus end interactions in S. cerevisiae. AB - Nuclear movement before karyogamy in eukaryotes is known as pronuclear migration or as nuclear congression in Saccharomyces cerevisiae. In this study, S. cerevisiae is used as a model system to study microtubule (MT)-dependent nuclear movements during mating. We find that nuclear congression occurs through the interaction of MT plus ends rather than sliding and extensive MT overlap. Furthermore, the orientation and attachment of MTs to the shmoo tip before cell wall breakdown is not required for nuclear congression. The MT plus end-binding proteins Kar3p, a class 14 COOH-terminal kinesin, and Bik1p, the CLIP-170 orthologue, localize to plus ends in the shmoo tip and initiate MT interactions and depolymerization after cell wall breakdown. These data support a model in which nuclear congression in budding yeast occurs by plus end MT capture and depolymerization, generating forces sufficient to move nuclei through the cytoplasm. This is the first evidence that MT plus end interactions from oppositely oriented organizing centers can provide the force for organelle transport in vivo. PMID- 16380441 TI - Single-channel properties of human NaV1.1 and mechanism of channel dysfunction in SCN1A-associated epilepsy. AB - Mutations in genes encoding neuronal voltage-gated sodium channel subunits have been linked to inherited forms of epilepsy. The majority of mutations (>100) associated with generalized epilepsy with febrile seizures plus (GEFS+) and severe myoclonic epilepsy of infancy (SMEI) occur in SCN1A encoding the NaV1.1 neuronal sodium channel alpha-subunit. Previous studies demonstrated functional heterogeneity among mutant SCN1A channels, revealing a complex relationship between clinical and biophysical phenotypes. To further understand the mechanisms responsible for mutant SCN1A behavior, we performed a comprehensive analysis of the single-channel properties of heterologously expressed recombinant WT-SCN1A channels. Based on these data, we then determined the mechanisms for dysfunction of two GEFS+-associated mutations (R1648H, R1657C) both affecting the S4 segment of domain 4. WT-SCN1A has a slope conductance (17 pS) similar to channels found in native mammalian neurons. The mean open time is approximately 0.3 ms in the 30 to -10 mV range. The R1648H mutant, previously shown to display persistent sodium current in whole-cell recordings, exhibited similar slope conductance but had an increased probability of late reopening and a subfraction of channels with prolonged open times. We did not observe bursting behavior and found no evidence for a gating mode shift to explain the increased persistent current caused by R1648H. Cells expressing R1657C exhibited conductance, open probability, mean open time, and latency to first opening similar to WT channels but reduced whole cell current density, suggesting decreased number of functional channels at the plasma membrane. In summary, our findings define single-channel properties for WT SCN1A, detail the functional phenotypes for two human epilepsy-associated sodium channel mutants, and clarify the mechanism for increased persistent sodium current induced by the R1648H allele. PMID- 16380442 TI - Intracellular pH modulates taste receptor cell volume and the phasic part of the chorda tympani response to acids. AB - The relationship between cell volume and the neural response to acidic stimuli was investigated by simultaneous measurements of intracellular pH (pHi) and cell volume in polarized fungiform taste receptor cells (TRCs) using 2',7'-bis-(2 carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) in vitro and by rat chorda tympani (CT) nerve recordings in vivo. CT responses to HCl and CO2 were recorded in the presence of 1 M mannitol and specific probes for filamentous (F) actin (phalloidin) and monomeric (G) actin (cytochalasin B) under lingual voltage clamp. Acidic stimuli reversibly decrease TRC pHi and cell volume. In isolated TRCs F-actin and G-actin were labeled with rhodamine phalloidin and bovine pancreatic deoxyribonuclease-1 conjugated with Alexa Fluor 488, respectively. A decrease in pHi shifted the equilibrium from F-actin to G-actin. Treatment with phalloidin or cytochalasin B attenuated the magnitude of the pHi-induced decrease in TRC volume. The phasic part of the CT response to HCl or CO2 was significantly decreased by preshrinking TRCs with hypertonic mannitol and lingual application of 1.2 mM phalloidin or 20 microM cytochalasin B with no effect on the tonic part of the CT response. In TRCs first treated with cytochalasin B, the decrease in the magnitude of the phasic response to acidic stimuli was reversed by phalloidin treatment. The pHi-induced decrease in TRC volume induced a flufenamic acid sensitive nonselective basolateral cation conductance. Channel activity was enhanced at positive lingual clamp voltages. Lingual application of flufenamic acid decreased the magnitude of the phasic part of the CT response to HCl and CO2. Flufenamic acid and hypertonic mannitol were additive in inhibiting the phasic response. We conclude that a decrease in pHi induces TRC shrinkage through its effect on the actin cytoskeleton and activates a flufenamic acid-sensitive basolateral cation conductance that is involved in eliciting the phasic part of the CT response to acidic stimuli. PMID- 16380443 TI - Proton sensing of CLC-0 mutant E166D. AB - CLC Cl- channels are homodimers in which each subunit has a proper pore and a (fast) gate. An additional slow gate acts on both pores. A conserved glutamate (E166 in CLC-0) is a major determinant of gating in CLC-0 and is crucially involved in Cl-/H+ antiport of CLC-ec1, a CLC of known structure. We constructed tandem dimers with one wild-type (WT) and one mutant subunit (E166A or E166D) to show that these mutations of E166 specifically alter the fast gate of the pore to which they belong without effect on the fast gate of the neighboring pore. In addition both mutations activate the common slow gate. E166A pores have a large, voltage-independent open probability of the fast gate (popen), whereas popen of E166D pores is dramatically reduced. Similar to WT, popen of E166D was increased by lowering pHint. At negative voltages, E166D presents a persistent inward current that is blocked by p-chlorophenoxy-acetic acid (CPA) and increased at low pHext. The pHext dependence of the persistent current is analogous to a similar steady inward current in WT CLC-0. Surprisingly, however, the underlying unitary conductance of the persistent current in E166D is about an order of magnitude smaller than that of the transient deactivating inward Cl- current. Collectively, our data support the possibility that the mutated CLC-0 channel E166D can assume two distinct open states. Voltage-independent protonation of D166 from the outside favors a low conductance state, whereas protonation from the inside favors the high conductance state. PMID- 16380444 TI - Regulation of connexin hemichannels by monovalent cations. AB - Opening of connexin hemichannels in the plasma membrane is highly regulated. Generally, depolarization and reduced extracellular Ca2+ promote hemichannel opening. Here we show that hemichannels formed of Cx50, a principal lens connexin, exhibit a novel form of regulation characterized by extraordinary sensitivity to extracellular monovalent cations. Replacement of extracellular Na+ with K+, while maintaining extracellular Ca2+ constant, resulted in >10-fold potentiation of Cx50 hemichannel currents, which reversed upon returning to Na+. External Cs+, Rb+, NH4+, but not Li+, choline, or TEA, exhibited a similar effect. The magnitude of potentiation of Cx50 hemichannel currents depended on the concentration of extracellular Ca2+, progressively decreasing as external Ca2+ was reduced. The primary effect of K+ appears to be a reduction in the ability of Ca2+, as well as other divalent cations, to close Cx50 hemichannels. Cx46 hemichannels exhibited a modest increase upon substituting Na+ with K+. Analyses of reciprocal chimeric hemichannels that swap NH2- and COOH-terminal halves of Cx46 and Cx50 demonstrate that the difference in regulation by monovalent ions in these connexins resides in the NH2-terminal half. Connexin hemichannels have been implicated in physiological roles, e.g., release of ATP and NAD+ and in pathological roles, e.g., cell death through loss or entry of ions and signaling molecules. Our results demonstrate a new, robust means of regulating hemichannels through a combination of extracellular monovalent and divalent cations, principally Na+, K+, and Ca2+. PMID- 16380445 TI - Polymorphisms in the IL10 but not in the IL1beta and IL4 genes are associated with inhibitor development in patients with hemophilia A. AB - The aim of the Malmo International Brother Study (MIBS) is to evaluate host genetic factors associated with the development of inhibitory antibodies in patients with hemophilia. Factor VIII gene mutations and genetic polymorphisms of the IL1beta, IL4, and IL10 genes, known to influence antibody production in autoimmune diseases, were analyzed in 164 patients (124 with severe, 26 with moderate, and 14 with mild disease) in 78 unrelated families with hemophilia A. Seventy-seven (47%) patients in 54 families had a history of inhibitors (57 high responding, 20 low responding). Inversions were found in 36 families (75 patients). There was no association between the development of inhibitor and the IL1beta Taq I RFLP alleles in exon 5 or the -590 C/T single nucleotide polymorphism (SNP) in the promoter region of IL4. There was, however, a strong association between an allele with 134 bp in one of the CA repeat microsatellites, IL10G, located in the promoter region of the IL10 gene, and the development of inhibitor (odds ratio [OR], 4.4; 95% confidence interval [95% CI], 2.1-9.5; P < .001). The association was consistent in the subgroup of families with severe hemophilia and inversions. IL10 is the first gene located outside the causative factor VIII gene mutation to be associated with inhibitor development. PMID- 16380446 TI - NF-kappaB is essential for the progression of KSHV- and EBV-infected lymphomas in vivo. AB - Activated NF-kappaB is a critical mechanism by which lymphoma cells infected by Epstein-Barr virus (EBV/HHV-4) and Kaposi sarcoma herpesvirus (KSHV/HHV-8) are protected from apoptotic stress. Selective pharmacologic inhibition of constitutive NF-kappaB activity induces apoptosis in KSHV- and EBV-infected lymphoma cells. In both tumor types, pharmacologic inhibition of NF-kappaB in vitro induced identical mitochondrially mediated apoptosis cascades. Assessment of gene regulation by microarray analysis revealed that the inhibition of NF kappaB in tumor cells results in the down-regulation of a distinct group of prosurvival genes, including cIAP-1, cIAP-2, cFLIP, and IL-6. Using EBV- and KSHV associated lymphomas in a murine system, we demonstrated that Bay 11-7082, a selective pharmacologic inhibitor of NF-kappaB, prevents or delays tumor growth and prolongs disease-free survival. Inhibition of NF-kappaB activity and tumor growth responses were further documented using a traceable reporter KSHV-positive cell line and in vivo imaging. These findings indicate that specific NF-kappaB regulated survival factors work cooperatively to protect KSHV- and EBV-infected lymphoma cells from apoptosis such that they promote the establishment and progression of KSHV- and EBV-associated lymphomas in mice. They also support the use of selective NF-kappaB inhibitors in the treatment of herpesvirus-associated lymphomas. PMID- 16380447 TI - Identification of cutaneous lymphocyte-associated antigen as sialyl 6-sulfo Lewis X, a selectin ligand expressed on a subset of skin-homing helper memory T cells. AB - We previously identified the carbohydrate determinant sialyl 6-sulfo Lewis X (Le(x)) as the major L-selectin ligand on high endothelial venules of peripheral lymph nodes. In this study, we examined the distribution of the sialyl 6-sulfo Le(x) determinant among peripheral lymphocytes. The determinant was expressed on a subset of helper memory T and NK cells. The helper memory T cells expressing sialyl 6-sulfo Le(x) were CD45RO(bright+) PSGL-1(high+) CCR4+ L-selectin+ CCR7+ but did not express alpha4beta7 integrin or CCR9, indicating that they were the skin-homing population of central memory T cells. The T-cell subset significantly expressed mRNA for 6-sulfotransferase HEC-GlcNAc6ST and fucosyltransferase Fuc-T VII, responsible for the synthesis of sialyl 6-sulfo Le(x). Characteristics of the T-cell population were similar to those previously described for cutaneous lymphocyte-associated antigen (CLA)-positive T cells defined by the HECA-452 or 2F3 antibody. The binding of the T-cell subset with the specific anti-sialyl 6 sulfo Le(x) antibody G152 was almost completely abrogated by HECA-452 or 2F3. Binding of recombinant E-, P-, and L-selectins to the T-cell subset was significantly inhibited by G152 and by HECA-452 antibodies. We propose that CLA, which is expressed without any activation stimuli on peripheral skin-homing helper memory T cells in healthy persons, is at least partly the sialyl 6-sulfo Le(x) determinant. PMID- 16380448 TI - Stem cell mobilization with G-CSF analogs: a rational approach to separate GVHD and GVL? AB - The separation of graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) remains the "holy grail" of allogeneic stem cell transplantation, and improvements are urgently needed to allow more effective therapy of malignant disease. The use of G-CSF-mobilized peripheral blood as a clinical stem cell source is associated with enhanced GVL effects without amplification of significant acute GVHD. Preclinical studies have demonstrated that G-CSF modulates donor T cell function before transplantation, promoting T(H)2 differentiation and regulatory T cell function. In addition, the expansion of immature antigen-presenting cells (APCs) and plasmacytoid dendritic cells (DCs) favors the maintenance of this pattern of T cell differentiation after transplantation. Although these patterns of T cell differentiation attenuate acute GVHD, they do not have an impact on the cytolytic pathways of the CD8(+) T cells that are critical for effective GVL. Recently, it has been demonstrated that modification of G-CSF, either by pegylation of the native cytokine or conjugation to Flt-3L, results in the expansion and activation of donor iNKT cells, which significantly augment CD8(+) T cell-mediated cytotoxicity and GVL effects after transplantation. Given that these cytokines also enhance the expansion of regulatory T cells and APCs, they further separate GVHD and GVL, offering potential clinical advantages for the transplant recipient. PMID- 16380449 TI - Fetomaternal cross talk in the placental vascular bed: control of coagulation by trophoblast cells. AB - Humans and rodents exhibit a peculiar type of placentation in which zygote derived trophoblast cells, rather than endothelial cells, line the terminal maternal vascular space. This peculiar aspect of the placental vasculature raises important questions about the relative contribution of fetal and maternal factors in the local control of hemostasis in the placenta and how these might determine the phenotypic expression of thrombophilia-associated complications of pregnancy. Using genomewide expression analysis, we identify a panel of genes that determine the ability of fetal trophoblast cells to regulate hemostasis at the fetomaternal interface. We show that spontaneous differentiation of trophoblast stem cells is associated with the acquisition of an endothelial cell-like thromboregulatory gene expression program. This program is developmentally regulated and conserved between mice and humans. We further show that trophoblast cells sense, via the expression of protease activated receptors, the presence of activated coagulation factors. Engagement of these receptors results in cell-type specific changes in gene expression. Our observations define candidate fetal genes that are potential risk modifiers of maternal thrombophilia-associated pregnancy complications and provide evidence that coagulation activation at the fetomaternal interface can affect trophoblast physiology altering placental function in the absence of frank thrombosis. PMID- 16380450 TI - Stress-induced cholinergic signaling promotes inflammation-associated thrombopoiesis. AB - To study the role of the stress-induced "readthrough" acetylcholinesterase splice variant, AChE-R, in thrombopoiesis, we used transgenic mice overexpressing human AChE-R (TgR). Increased AChE hydrolytic activity in the peripheral blood of TgR mice was associated with increased thrombopoietin levels and platelet counts. Bone marrow (BM) progenitor cells from TgR mice presented an elevated capacity to produce mixed (GEMM) and megakaryocyte (Mk) colonies, which showed intensified labeling of AChE-R and its interacting proteins RACK1 and PKC. When injected with bacterial lipopolysaccharide (LPS), parent strain FVB/N mice, but not TgR mice, showed reduced platelet counts. Therefore, we primed human CD34+ cells with the synthetic ARP26 peptide, derived from the cleavable C-terminus of AChE-R prior to transplantation, into sublethally irradiated NOD/SCID mice. Engraftment of human cells (both CD45+ and CD41+ Mk) was significantly increased in mice that received ARP26-primed CD34+ human cells versus mice that received fresh nonprimed CD34+ human cells. Moreover, ARP26 induced polyploidization and proplatelet shedding in human MEG-01 promegakaryotic cells, and human platelet engraftment increased following ex vivo expansion of ARP26-treated CD34+ cells as compared to cells expanded with thrombopoietin and stem cell factor. Our findings implicate AChE-R in thrombopoietic recovery, suggesting new therapeutic modalities for supporting platelet production. PMID- 16380452 TI - Epithelial-specific transcription factor ESE-3 is involved in the development of monocyte-derived DCs. AB - Dendritic cells (DCs) are recognized as the most potent antigen-presenting cells of the immune system with the unique ability to initiate and maintain primary immune responses. In order to better characterize the functional and phenotypic features of DCs, a subtractive cDNA library to identify differentially expressed genes in monocyte-derived DCs (MDCs) was constructed. Using this approach, we found that the epithelial transcription factor ESE-3, which was previously shown to be exclusively expressed in cells of epithelial origin, is differentially expressed in MDCs. This was further confirmed by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analyses. The expression of ESE-3 is up regulated upon maturation of MDCs and inhibited by treating the cells with IL-10 or IFN-gamma. Knockdown experiments using siRNA suggest that ESE-3 plays an important role during MDC development. Our results might help to improve the phenotypic characterization of DCs and lead to a better understanding of the cellular mechanisms involved in antigen presentation and T-cell stimulation. PMID- 16380451 TI - A global role for EKLF in definitive and primitive erythropoiesis. AB - Erythroid Kruppel-like factor (EKLF, KLF1) plays an important role in definitive erythropoiesis and beta-globin gene regulation but failure to rectify lethal fetal anemia upon correction of globin chain imbalance suggested additional critical EKLF target genes. We employed expression profiling of EKLF-null fetal liver and EKLF-null erythroid cell lines containing an inducible EKLF-estrogen receptor (EKLF-ER) fusion construct to search for such targets. An overlapping list of EKLF-regulated genes from the 2 systems included alpha-hemoglobin stabilizing protein (AHSP), cytoskeletal proteins, hemesynthesis enzymes, transcription factors, and blood group antigens. One EKLF target gene, dematin, which encodes an erythrocyte cytoskeletal protein (band 4.9), contains several phylogenetically conserved consensus CACC motifs predicted to bind EKLF. Chromatin immunoprecipitation demonstrated in vivo EKLF occupancy at these sites and promoter reporter assays showed that EKLF activates gene transcription through these DNA elements. Furthermore, investigation of EKLF target genes in the yolk sac led to the discovery of unexpected additional defects in the embryonic red cell membrane and cytoskeleton. In short, EKLF regulates global erythroid gene expression that is critical for the development of primitive and definitive red cells. PMID- 16380453 TI - Semaphorin-3A is expressed by tumor cells and alters T-cell signal transduction and function. AB - An important aspect of tumor progression is the ability of cancer cells to escape detection and clearance by the immune system. Recent studies suggest that several tumors express soluble factors interfering with the immune response. Here, we show that semaphorin-3A (Sema-3A), a secreted member of the semaphorin family involved in axonal guidance, organogenesis, and angiogenesis, is highly expressed in several tumor cells. Conditioned media of Sema-3A-transfected COS-7 cells or human recombinant Sema-3A inhibited primary human T-cell proliferation and cytokines production under anti-CD3 plus anti-CD28 stimulating conditions. Sema 3A also inhibited the activation of nonspecific cytotoxic activity in mixed lymphocyte culture (MLC), as measured against K-562 cells. In contrast, suppression of Sema-3A in tumor cells with a small interfering RNA (siRNA) augmented T-cell activation. The inhibitory effect of Sema-3A in T cells is mediated by blockade of Ras/mitogen-activated protein kinase (MAPK) signaling pathway. The presence of Sema-3A increased the activation of the Ras family small GTPase Rap1 and introduction of the dominant-negative mutant of Rap1 (Rap1N17) blunted the immunoinhibitory effects of Sema-3A. These results suggest that Sema 3A inhibits primary T-cell activation and imply that it can contribute to the T cell dysfunction in the tumor microenvironment. PMID- 16380454 TI - Conditioning including antithymocyte globulin followed by unmanipulated HLA mismatched/haploidentical blood and marrow transplantation can achieve comparable outcomes with HLA-identical sibling transplantation. AB - The outcomes of 293 patients with leukemia undergoing HLA-identical sibling (n = 158) or related HLA-mismatched (n = 135) hematopoietic cell transplantation (HCT) performed during the same time period were compared. Patients received BUCY2 in HLA-identical sibling HCT or BUCY2 + ATG in mismatched HCT as conditioning regimens, followed by unmanipulated marrow and/or peripheral blood (PB) transplantation. All patients achieved full engraftment. The cumulative incidences of grades II to IV acute graft-versus-host disease (aGVHD) in the matched and mismatched cohorts were 32% (CI, 25%-39%) versus 40% (CI, 32%-48%, P = .13), respectively, with the relative risk (RR) = 0.64 (95% CI, 0.43-0.94), P = .02. The incidence of chronic GVHD did not differ significantly between the cohorts (P = .97). Two-year incidences of treatment-related mortality and relapse for matched versus mismatched were 14% (range, 9%-20%) versus 22% (range, 15% 29%) with P = .10 and 13% (range, 8%-19%) versus 18% (range, 10%-27%) with P = .40, respectively. Two-year adjusted leukemia-free survival (LFS) and overall survival were 71% (range, 63%-78%) versus 64% (range, 54%-73%) with P = .27 and 72% (range, 64%-79%) versus 71% (range, 62%-77%) with P = .72, respectively. Multivariate analyses showed that only advanced disease stage and a diagnosis of acute leukemia had increased risk of relapse, treatment failure, and overall mortality. In summary, HCT performed with related HLA-mismatched donors is a feasible approach with acceptable outcomes. PMID- 16380455 TI - AML1-ETO rapidly induces acute myeloblastic leukemia in cooperation with the Wilms tumor gene, WT1. AB - AML1-ETO, a chimeric gene frequently detected in acute myelogenous leukemia (AML), inhibits the differentiation of myeloid progenitors by suppressing genes associated with myeloid differentiation and increases the replating ability of clonogenic myeloid progenitors. However, AML1-ETO alone cannot induce AML and thus additional genetic events are required for the onset of AML. The Wilms tumor gene (WT1), which has been identified as the gene responsible for Wilms tumor, is expressed at high levels in almost all human leukemias. In this study, we have generated transgenic mice (WT1-Tg) that overexpress WT1 in hematopoietic cells to investigate the effects of WT1 on AML1-ETO-associated leukemogenesis. AML1-ETO transduced bone marrow (BM) cells from WT1-Tg mice exhibited inhibition of myeloid differentiation at more immature stages and higher in vitro colony forming ability compared with AML1-ETO-transduced BM cells from wild-type mice. Most importantly, all of the mice that received a transplant of AML1-ETO transduced BM cells from the WT1-Tg mice rapidly developed AML. These results demonstrate that AML1-ETO may exert its leukemogenic function in cooperation with the expression of WT1. PMID- 16380456 TI - Three-dimensional architecture of inner medullary vasa recta. AB - The manner in which vasa recta function and contribute to the concentrating mechanism depends on their three-dimensional relationships to each other and to tubular elements. We have examined the three-dimensional architecture of vasculature relative to tubular structures in the central region of rat kidney inner medulla from the base through the first 3 mm by combining immunohistochemistry and semiautomated image acquisition techniques with graphical modeling software. Segments of descending vasa recta (DVR), ascending vasa recta (AVR), descending thin limb (DTL), ascending thin limb (ATL), and collecting duct (CD) were identified with antibodies against segment-specific proteins associated with solute and water transport (urea channel B, PV-1, aquaporin-1, ClC-K1, aquaporin-2, respectively) by immunofluorescence. Results indicate: 1) DVR, like DTLs, are excluded from CD clusters that we have previously shown to be the organizing element for the inner medulla; 2) AVR, like ATLs, are nearly uniformly distributed transversely across the entire inner medulla outside of and within CD clusters; 3) DVR and AVR outside CD clusters appear to be sufficiently juxtaposed to permit good countercurrent exchange; 4) within CD clusters, about four AVR closely abut each CD, surrounding it in a highly symmetrical fashion; and 5) AVR abutting each CD and ATLs within CD clusters form repeating nodal interstitial spaces bordered by a CD on one side, one or more ATLs on the opposite side, and one AVR on each of the other two sides. These relationships may be highly significant for both establishing and maintaining the inner medullary osmotic gradient. PMID- 16380457 TI - Endothelin-1 stimulates NO production and inhibits cAMP accumulation in rat inner medullary collecting duct through independent pathways. AB - Endothelin-1 (ET-1) inhibition of vasopressin (AVP)-stimulated cAMP accumulation in the collecting duct has been hypothesized to be mediated, at least in part, by nitric oxide (NO). To examine this, the effect of ET-1 on NO production by acutely isolated rat inner medullary collecting duct (IMCD) cell suspensions and the role of NO in mediating ET-1 effects on AVP-stimulated cAMP accumulation were studied. ET-1 dose dependently (first evident at 100 pM ET-1) increased IMCD NO production as determined by DAF-FM fluorescence. ET(B) receptor (BQ-788), but not ET(A) receptor (BQ-123), antagonism blocked this effect. Nonspecific NO synthase (NOS) inhibitors [N(G)-nitro-L-arginine methyl ester (L-NAME) or N(G)-monomethyl L-arginine] or NOS-1 inhibitors (SMTC or VNIO) inhibited the ET-1 response, whereas NOS-2 or NOS-3 inhibitors (L-NAA or 1400W) were ineffective. ET-1 also increased cGMP accumulation. ET-1 caused a 35% reduction in AVP-stimulated cAMP levels; however, this response was not affected by L-NAME or SMTC. The addition of L-arginine, NADPH, tetrahydrobiopterin, or tempol (to reduce superoxide dependent conversion of NO to peroxynitrate) did not affect the response. NO donors (SNAP or spermine NONOate), at concentrations that stimulated DAF-FM fluorescence and increased cGMP levels, did not alter AVP-stimulated cAMP accumulation in the IMCD cell suspensions. In conclusion, ET-1 stimulates IMCD NO production through activation of the ET(B) receptor and NOS-1. However, neither ET-1-mediated NO production nor NO donors inhibit AVP-stimulated cAMP accumulation, indicating that NO does not mediate ET-1 inhibition of cAMP production by the IMCD. PMID- 16380458 TI - Molecular characterization of the mercurial sensitivity of a frog urea transporter (fUT). AB - The amphibian urea transporter (fUT) shares many properties with the mammalian urea transporters (UT) derived from UT-A and UT-B genes. The transport of urea by fUT is inhibited by the mercurial agent p-chloromercuribenzenesulfonic acid (pCMBS). We found that in oocytes expressing cRNA encoding fUT, a 5-min preincubation in 0.5 mM mercury chloride (HgCl2) also significantly reduced urea uptake. The transport of urea by fUT was rendered mercury (Hg2+) insensitive by mutating either of the residues C185 or H187, both of which lie within the M-I region (close to the hypothetical UT pore). In oocytes expressing a mixture of the C185 and H187 mutants, Hg2+ sensitivity was reestablished. The transport of urea by the mouse UTs mUT-A2 and mUT-A3 was not sensitive to Hg2+. Introducing cysteine residues analogous to that mutated in fUT into mUT-A2 or mUT-A3 did not induce Hg2+ sensitivity. Additionally, introducing the double cysteine, histidine mutations into mUT-A2 or mUT-A3 still did not induce Hg2+ sensitivity, indicating that a region outside of the M-I region also contributes to the Hg2+-induced block of fUT. Using a series of chimeras formed between UT-A3 and fUT, we found that as well as C185 and H187, residues within the COOH terminal of fUT determine Hg2+ sensitivity, and we propose that differences in the folding of this region between fUT and mUT-A2/mUT-A3 allow access of Hg2+ to the fUT channel pore. PMID- 16380459 TI - Nitric oxide stimulates cyclooxygenase-2 in cultured cTAL cells through a p38 dependent pathway. AB - To examine the interaction of nitric oxide (NO) and cyclooxygenase (COX-2) and the signaling pathway involved, primary cultured rabbit cortical thick ascending limb (cTAL) were used. In these cells, immunoreactive COX-2 and vasodilatory prostaglandins were increased by a NO donor, S-nitros-N-acetylpenicillamine (SNAP; 2.5 +/- 0.3-fold control, n = 6, P < 0.01). SNAP increased expression of phosphorylated p38 (pp38; 2.4 +/- 0.3-fold control; n = 5; P < 0.01), which was inhibited by the p38 inhibitor SB-203580 (1.3 +/- 0.1-fold control, n = 5, P < 0.01). SB-203580 inhibited SNAP-induced COX-2 expression [1.4 +/- 0.2-fold control, n = 6, not significant (NS) vs. control] and levels of PGE2 significantly. In cTAL cells transfected with a luciferase reporter driven by the wild-type mouse COX-2 promoter, SNAP stimulated luciferase activity, which was reversed by SB-203580 (control vs. SNAP vs. SNAP + SB-203580: 1.4 +/- 0.2-, 8.3 +/- 1.4-, and 0.4 +/- 0.1-fold control, respectively, n = 4, P < 0.01). Electrophoretic mobility shift assay indicated that SNAP stimulated nuclear factor (NF)-kappaB binding activity in cTAL that was also inhibited by the p38 inhibitor. SNAP was not able to stimulate a mutant COX-2 promoter construct that is not activated by NF-kappaB (0.9 +/- 0.1, 1.2 +/- 0.1, and 1.0 +/- 0.2 respectively, n = 4, NS). Low chloride increased COX-2 expression (2.7 +/- 0.4 fold control, n = 6, P < 0.01) and pp38 expression (2.8 +/- 0.3-fold; n = 5, P < 0.01), which were reversed by the specific NO synthase (NOS) inhibitor 7 nitroindazole. Administration of a low-salt diet increased immunoreactive COX-2 and neuronal NOS (nNOS) in the macula densa and surrounding cTAL of kidneys of wild-type mice but did not significantly elevate COX-2 expression in nNOS-/- mice. In summary, these studies indicate that, in cTAL, NO can increase COX-2 expression in cTAL and macula densa through p38-dependent signaling pathways via activation of NF-kappaB. PMID- 16380460 TI - Role of fibrillin-1 in hypertensive and diabetic glomerular disease. AB - The microfibrillar protein fibrillin-1 is a component of the mesangial matrix. Defects in fibrillin-1 predisposes individuals to vascular damage in Marfan syndrome, but the role of fibrillin-1 in kidney disease is unknown. We hypothesized that fibrillin-1 is involved in hypertensive or diabetic glomerular disease. DOCA-salt hypertension or streptozotocin (STZ) diabetes led to a significant increase in glomerular fibrillin-1 deposition. To test the functional role of fibrillin-1, DOCA hypertension and STZ diabetes were induced in mice homozygous for a mutation leading to a fivefold lower expression of fibrillin-1 (mgR/mgR). Untreated male mgR/mgR mice usually die from aortic dissection during the first 4 mo of life. All DOCA-treated mgR/mgR mice died within 2 wk after onset of DOCA treatment. DOCA-treated heterozygous (mgR/+) and their wild-type littermates displayed similar blood pressure levels, but albuminuria was significantly lower in mgR/+ than in wild-type mice after DOCA treatment. Similarly, STZ diabetic mgR/mgR and mgR/+ developed lower albuminuria than wild type mice despite higher blood glucose levels in mgR/mgR and mgR/+ compared with wild-type mice. Blood pressure, blood glucose, and albuminuria did not differ among untreated mgR/mgR, mgR/+, and wild-type mice, respectively. In diabetic mgR/+ and mgR/mgR, but not in wild-type mice, an induction of glomerular decorin expression was observed. Thus underexpression of fibrillin-1 predisposes individuals to lethal aortic dissection in the presence of hypertension. On the other hand, albuminuria as a parameter of microvascular damage in hypertension and diabetes was ameliorated in fibrillin-1-underexpressing mice, possibly due to a compensatory upregulation of decorin. We conclude that fibrillin-1 may contribute to glomerular damage in hypertensive and diabetic kidney disease. PMID- 16380461 TI - Intracellular ANG II induces cytosolic Ca2+ mobilization by stimulating intracellular AT1 receptors in proximal tubule cells. AB - Intracellular ANG II induces biological effects in nonrenal cells, but it is not known whether it plays a physiological role in renal proximal tubule cells (PTCs). PTCs express angiotensinogen, renin, and angiotensin-converting enzyme mRNAs, suggesting the presence of high levels of intracellular ANG II. We determined if microinjection of ANG II directly in single PTCs increases intracellular calcium concentration ([Ca2+]i) and, if so, elucidated the cellular mechanisms involved. Changes in [Ca2+]i responses were studied by fluorescence imaging using the Ca2+ indicator fluo 3. ANG II (1 nM) was microinjected directly in the cells, whereas cell-surface angiotensin type 1 (AT1) receptors were blocked by losartan (10 microM). When ANG II (1 nM) was added to the perfusate, there was a marked increase in [Ca2+]i that was blocked by extracellular losartan. With losartan in the perfusate, intracellular microinjection of ANG II elicited a robust increase in cytoplasmic [Ca2+]i that peaked at 30 s (basal: 2.2 +/- 0.3 vs. ANG II: 14.9 +/- 0.4 relative fluorescence units; P < 0.01). Chelation of extracellular Ca2+ with EGTA (2 mM) did not alter microinjected ANG II-induced [Ca2+]i responses (Ca2+ free + ANG II: 12.3 +/- 2.6 relative fluorescence units, not significant vs. ANG II); however, pretreatment with thapsigargin to deplete intracellular Ca2+ stores or with U-73122 to inhibit phospholipase C (1 microM each) markedly attenuated microinjected ANG II-induced [Ca2+]i responses. Combined microinjection of ANG II and losartan abolished [Ca2+]i responses, whereas a combination of ANG II and PD-123319 had no effect. These data demonstrate for the first time that direct microinjection of ANG II in single PTCs increases [Ca2+]i by stimulating intracellular AT1 receptors and releases Ca2+ from intracellular stores, suggesting that intracellular ANG II may play a physiological role in PTC function. PMID- 16380462 TI - NFAT regulates calcium-sensing receptor-mediated TNF production. AB - Because nuclear factor of activated T cells (NFAT) has been implicated in TNF production as well as osmoregulation and salt and water homeostasis, we addressed whether calcium-sensing receptor (CaR)-mediated TNF production in medullary thick ascending limb (mTAL) cells was NFAT dependent. TNF production in response to addition of extracellular Ca(2+) (1.2 mM) was abolished in mTAL cells transiently transfected with a dominant-negative CaR construct (R796W) or pretreated with the phosphatidylinositol phospholipase C (PI-PLC) inhibitor U-73122. Cyclosporine A (CsA), an inhibitor of the serine/threonine phosphatase calcineurin, and a peptide ligand, VIVIT, that selectively inhibits calcineurin-NFAT signaling, also prevented CaR-mediated TNF production. Increases in calcineurin activity in cells challenged with Ca(2+) were inhibited after pretreatment with U-73122 and CsA, suggesting that CaR activation increases calcineurin activity in a PI-PLC dependent manner. Moreover, U-73122, CsA, and VIVIT inhibited CaR-dependent activity of an NFAT construct that drives expression of firefly luciferase in transiently transfected mTAL cells. Collectively, these data verify the role of calcineurin and NFAT in CaR-mediated TNF production by mTAL cells. Activation of the CaR also increased the binding of NFAT to a consensus oligonucleotide, an effect that was blocked by U-73122 and CsA, suggesting that a calcineurin- and NFAT-dependent pathway increases TNF production in mTAL cells. This mechanism likely regulates TNF gene transcription as U-73122, CsA, and VIVIT blocked CaR dependent activity of a TNF promoter construct. Elucidating CaR-mediated signaling pathways that regulate TNF production in the mTAL will be crucial to understanding mechanisms that regulate extracellular fluid volume and salt balance. PMID- 16380463 TI - AT1 receptor-activated signaling mediates angiotensin IV-induced renal cortical vasoconstriction in rats. AB - Angiotensin IV (ANG IV), an active ANG II fragment, has been shown to induce systemic and renal cortical effects by binding to ANG IV (AT(4)) receptors and activating unique signaling transductions unrelated to classical type 1 (AT(1)) or type 2 (AT(2)) receptors. We tested whether ANG IV exerts systemic and renal cortical effects on blood pressure, renal microvascular smooth muscle cells (VSMCs), and glomerular mesangial cells (MC) and, if so, whether AT(1) receptor activated signaling is involved. In anesthetized rats, systemic infusion of ANG II, ANG III, or ANG IV (0.01, 0.1, and 1.0 nmol.kg(-1).min(-1) iv) caused dose dependent increases in mean arterial pressure (MAP) and decreases in renal cortical blood flow (CBF; P < 0.01). ANG II also induced dose-dependent reductions in renal medullary blood flow (P < 0.01), whereas ANG IV did not. ANG IV-induced pressor and renal cortical vasoconstriction were completely abolished by AT(1) receptor blockade with losartan (5 mg/kg iv; P < 0.05). When ANG IV (1 nmol.kg(-1).min(-1)) was infused directly in the renal artery, CBF was reduced by >30%, and the response was also blocked by losartan (P < 0.01). In the renal cortex, unlabeled ANG IV displaced (125)I-labeled [Sar(1),Ile(8)]ANG II binding, whereas unlabeled ANG II (10 microM) inhibited (125)I-labeled Nle(1)-ANG IV (AT(4)) binding in a concentration-dependent manner (P < 0.01). In freshly isolated renal VSMCs, ANG IV (100 nM) increased intracellular Ca(2+) concentration, and the effect was blocked by losartan and U-73122, a selective inhibitor of phospholipase C/inositol trisphosphate/Ca(2+) signaling (1 microM). In cultured rat MCs, ANG IV (10 nM) induced mitogen-activated protein kinase extracellular/signal-regulated kinase 1/2 phosphorylation via AT(1) receptor- and phospholipase C-activated signaling. These results suggest that, at nanomolar concentrations, ANG IV can increase MAP and induce renal cortical effects by interacting with AT(1) receptor-activated signaling. PMID- 16380464 TI - Angiotensin II AT2 receptors inhibit proximal tubular Na+-K+-ATPase activity via a NO/cGMP-dependent pathway. AB - Angiotensin II AT2 receptors act as a functional antagonist for the AT1 receptors in various tissues. We previously reported that activation of the renal AT2 receptors promotes natriuresis and diuresis; however, the mechanism is not known. The present study was designed to investigate whether activation of AT2 receptors affects the activity of Na+-K+-ATPase (NKA), an active tubular sodium transporter, in the proximal tubules isolated from Sprague-Dawley rats. The AT2 receptor agonist CGP-42112 (10(-10)-10(-7) M) produced a dose-dependent inhibition of NKA activity (9-38%); the inhibition was attenuated by the presence of the AT2 receptor antagonist PD-123319 (1 microM), suggesting the involvement of the AT2 receptors. The AT1 receptor antagonist losartan (1 microM) did not affect the CGP-42112 (100 nM)-induced inhibition of NKA activity. The presence of guanylyl cyclase inhibitor ODQ (10 microM) and the nitric oxide (NO) synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME; 100 microM) abolished the CGP-42112 (100 nM)-induced NKA inhibition. ANG II (100 nM), in the presence of losartan, significantly inhibited NKA activity; the inhibition was attenuated by PD-123319. CGP-42112 also, in a dose-dependent manner, stimulated NO production (approximately 0-230%) and cGMP accumulation (approximately 25-100%). The CGP-42112 (100 nM)-induced NO and cGMP increases were abolished by the AT2 receptor antagonist PD-123319, ODQ, and L-NAME. The data suggest that the activation of the AT2 receptor via stimulation of the NO/cGMP pathway causes inhibition of NKA activity in the proximal tubules. This phenomenon provides a plausible mechanism responsible for the AT2 receptor-mediated natriuresis diuresis in rodents. PMID- 16380465 TI - Temporal expression profile and distribution pattern indicate a role of connective tissue growth factor (CTGF/CCN-2) in diabetic nephropathy in mice. AB - Connective tissue growth factor (CTGF) is overexpressed in diabetic nephropathy (DN) and has therefore been implicated in its pathogenesis. The objective of the present study was to determine the tissue distribution of increased CTGF expression and the relationship of plasma, urinary, and renal CTGF levels to the development and severity of DN. We studied the relationship between CTGF and renal pathology in streptozotocin (STZ)-induced diabetes in C57BL/6J mice. Diabetic and age-matched control mice were killed after 1, 2, 4, and 9 wk of diabetes. In addition, key parameters of diabetes and DN were analyzed in 10-mo old diabetic ob/ob mice and their ob/+ littermates. STZ-induced diabetic mice showed a significantly increased urinary albumin excretion after 1 wk and increased mesangial matrix score after 2 wk. Increased renal fibronectin, fibronectin ED-A, and collagen IValpha1 expression, as well as elevated plasma creatinine levels, were observed after 9 wk. After 2 wk, CTGF mRNA was upregulated threefold in the renal cortex. By 9 wk, CTGF mRNA was also increased in the heart and liver. In contrast, transforming growth factor-beta1 mRNA content was significantly increased only in the kidney by 9 wk. Renal CTGF expression was mainly localized in podocytes and parietal glomerular epithelial cells, and less prominent in mesangial cells. In addition, plasma CTGF levels and urinary CTGF excretion were increased in diabetic mice. Moreover, albuminuria strongly correlated with urinary CTGF excretion (R = 0.83, P < 0.0001). Increased CTGF expression was also demonstrated in type 2 diabetic ob/ob mice, which points to a causal relationship between diabetes and CTGF and thus argues against a role of STZ in this process. The observed relationship of podocyte and urinary CTGF to markers of DN suggests a pathogenic role of CTGF in the development of DN. PMID- 16380468 TI - Intervention of the renal disaster relief task force (RDRTF) in the Kashmir earthquake. PMID- 16380466 TI - Megalin binds and internalizes angiotensin-(1-7). AB - Megalin is a multiligand receptor heavily involved in protein endocytosis. We recently demonstrated that megalin binds and mediates internalization of ANG II. Although there is a strong structural resemblance between ANG II and ANG-(1-7), their physiological actions and their affinity for the angiotensin type 1 receptor (AT(1)R) are dissimilar. Therefore, the hypothesis of the present work was to test whether megalin binds and internalizes ANG-(1-7). The uptake of ANG (1-7) was determined by exposure of confluent monolayers of BN/MSV cells (a model representative of the yolk sac epithelium) to fluorescently labeled ANG-(1-7) (100 nM) and measurement of the amount of cell-associated fluorescence after 4 h by flow cytometry. Anti-megalin antisera and an AT(1)R blocker (olmesartan) were used to interfere with uptake via megalin and the AT(1)R, respectively. ANG-(1-7) uptake was prevented by anti-megalin antisera (63%) to a higher degree than olmesartan (13%) (P < 0.001). In analysis by flow cytometry of binding experiments performed in brush-border membrane vesicles isolated from kidneys of CD-1 mice, anti-megalin antisera interfered with ANG-(1-7) binding more strongly than olmesartan (P < 0.05 against positive control). Interactions of megalin with ANG-(1-7) at a molecular level were studied by surface plasmon resonance, demonstrating that ANG-(1-7) binds megalin dose and time dependently and with an affinity similar to ANG II. These results show that the scavenger receptor megalin binds and internalizes ANG-(1-7). PMID- 16380470 TI - The network of glucokinase-expressing cells in glucose homeostasis and the potential of glucokinase activators for diabetes therapy. AB - The glucose-phosphorylating enzyme glucokinase has structural, kinetic, and molecular genetic features that are ideal for its primary role as glucose sensor in a network of neuro/endocrine sentinel cells that maintain glucose homeostasis in many vertebrates including humans. The glucokinase-containing, insulin producing beta-cells of the pancreas take the prominent lead in this network, functioning in the aggregate as the master gland. The beta-cells are also conceptualized as the prototype for all other glucose sensor cells, which determines our current understanding of many extrapancreatic glucose sensors. About 99% of the enzyme resides, however, in the hepato-parenchymal cells and serves its second role in a high-capacity process of blood glucose clearance. Two examples strikingly illustrate how pivotal a position glucokinase has in the regulation of glucose metabolism: 1) activating and inactivating mutations of the enzyme cause hypo- and hyperglycemia syndromes in humans described collectively as "glucokinase disease" and fully explained by the glucose sensor paradigm, and 2) glucokinase activator drugs (GKAs) have been discovered that bind to an allosteric site and increase the kcat and lower the glucose S(0.5) of the enzyme. GKAs enhance glucose-stimulated insulin release from pancreatic islets and glucose disposition by the liver. They are now intensively explored to develop a novel treatment for diabetes. Future biophysical, molecular, genetic, and pharmacological studies hold much promise to unravel the evolving complexity of the glucokinase glucose sensor system. PMID- 16380471 TI - Noninvasive diagnosis of focal hyperinsulinism of infancy with [18F]-DOPA positron emission tomography. AB - Congenital hyperinsulinism of infancy (CHI) is characterized by severe hypoglycemia due to dysregulated insulin secretion, associated with either focal or diffuse pathology of the endocrine pancreas. The focal condition is caused by a paternally inherited mutation in one of the genes encoding the subunits of the beta-cell ATP-sensitive potassium channel (SUR1/ABCC8 or Kir6.2/KCNJ11) and somatic loss of maternal 11p15 alleles within the affected area. Until now, preoperative diagnostics have relied on technically demanding and invasive catheterization techniques. We evaluated the utility of fluorine-18 l-3,4 dihydroxyphenylalanine ([(18)F]-DOPA) positron emission tomography (PET) to identify focal pancreatic lesions in 14 CHI patients, 11 of which carried mutations in the ABCC8 gene (age 1-42 months). To reduce bias in PET image interpretation, quantitative means for evaluation of pancreatic [(18)F]-DOPA uptake were established. Five patients had a visually apparent focal accumulation of [(18)F]-DOPA and standardized uptake value (SUV) >50% higher (mean 1.8-fold) than the maximum SUV of the unaffected part of the pancreas. When these patients were operated on, a focus of 4-5 x 5-8 mm matching with the PET scan was found, and all were normoglycemic after resection of the focus. The remaining nine patients had diffuse accumulation of [(18)F]-DOPA in the pancreas (SUV ratio <1.5). Diffuse histology was verified in four of these, and pancreatic catheterization was consistent with diffuse pathology in four cases. In conclusion, [(18)F]-DOPA PET is a promising noninvasive method for the identification and localization of the focal form of CHI. PMID- 16380472 TI - Y2Y4 receptor double knockout protects against obesity due to a high-fat diet or Y1 receptor deficiency in mice. AB - Neuropeptide Y receptors are critical regulators of energy homeostasis, but the functional interactions and relative contributions of Y receptors and the environment in this process are unknown. We measured the effects of an ad libitum diet of normal or high-fat food on energy balance in mice with single, double, or triple deficiencies of Y1, Y2, or Y4 receptors. Whereas wild-type mice developed diet-induced obesity, Y2Y4 double knockouts did not. In contrast, Y1 knockout or Y1Y2 or Y1Y4 receptor double knockout mice developed an exacerbated diet-induced obesity syndrome. Remarkably, the antiobesity effect of Y2Y4 deficiency was stronger than the obesogenic effect of Y1 deficiency, since Y1Y2Y4 triple knockouts did not develop obesity on the high-fat diet. Resistance to diet induced obesity in Y2Y4 knockouts was associated with reduced food intake and improved glucose tolerance in the absence of changes in total physical activity. Fecal concentration of free fatty acids was significantly increased in Y2Y4 knockouts in association with a significantly reduced bile acid pool and marked alterations in intestinal morphology. In addition, hypothalamic proopiomelanocortin expression was decreased in diet-induced obesity (in both wild-type and Y1 receptor knockout mice) but not in obesity-resistant Y2Y4 receptor knockout mice fed a high-fat diet. Therefore, deletion of Y2 and Y4 receptors synergistically protects against diet-induced obesity, at least partially via changes in food intake and hypothalamic proopiomelanocortin expression. PMID- 16380473 TI - Costimulation blockade of both inducible costimulator and CD40 ligand induces dominant tolerance to islet allografts and prevents spontaneous autoimmune diabetes in the NOD mouse. AB - Costimulation blockade is a promising strategy for preventing allograft rejection and inducing tolerance. Using a fully allogeneic mouse model, we tested the effectiveness of the combined blockade of the CD40 ligand and the inducible costimulator (ICOS) on islet allograft survival and in the prevention of autoimmune diabetes in the NOD mouse. Recipients treated with blocking monoclonal antibodies (mAbs) to ICOS and the CD40 ligand had significant prolongation of graft survival, with 26 of 28 functioning for >200 days. Long-term engrafted mice maintained antidonor proliferative and cytotoxic responses, but donor-specific immunization did not induce graft rejection, and challenge with second, same donor but not third-party grafts resulted in long-term acceptance. The immunohistology of tolerant grafts demonstrated the presence of CD4(+)CD25(+) T cells expressing Foxp3, and islet/kidney composite grafts from tolerant mice, but not from mice lacking lymphocytes, were accepted indefinitely when transplanted into naive B6 mice, suggesting that recipient T-cells were necessary to generate dominant tolerance. Combined anti-ICOS and anti-CD40 ligand mAb therapy also prevented diabetes in NOD mice, with only 11% of treated recipients developing diabetes compared with 75% of controls. These data demonstrate that the blockade of CD40 ligand and ICOS signaling induces islet allograft tolerance involving a dominant mechanism associated with intragraft regulatory cells and prevents autoimmune diabetes in NOD mice. PMID- 16380474 TI - Natural killer T-cells participate in rejection of islet allografts in the liver of mice. AB - A role of natural killer T (NKT) cells in transplant rejection remains unknown. Here, we determined whether NKT cells participate in rejection of islet allografts, using NKT cell-deficient mice. Survival of islet allografts in streptozotocin-induced diabetic CD1d(-/-) mice or Valpha14 NKT cell(-/-) mice was significantly prolonged without immunosuppression when grafted into the liver, but not beneath the kidney capsule, compared with wild-type mice. Acceptance of intrahepatic islet allografts was achieved in CD1d(-/-) mice by a subtherapeutic dose of rapamycin, which was abrogated in conjunction with the transfer of hepatic mononuclear cells from wild-type, but not from CD1d(-/-), mice at islet transplantation. The second islet grafts from a donor-specific, but not from a third-party, strain in CD1d(-/-) mice bearing functional islet allografts were accepted without immunosuppression at 120 days after the initial transplantation. These findings demonstrate that NKT cells play a significant role in rejection of islet allografts in the liver of mice, but that NKT cells are not essential for induction of donor-specific unresponsiveness in this model. The current study indicates that NKT cells might be considered as a target for intervention to prevent islet allograft rejection when the liver is the site of transplantation. PMID- 16380475 TI - Rapamycin and interleukin-10 treatment induces T regulatory type 1 cells that mediate antigen-specific transplantation tolerance. AB - Islet transplantation is a cure for type 1 diabetes, but its potential is limited by the need for constant immunosuppression. One solution to this problem is the induction of transplantation tolerance mediated by T regulatory cells. T regulatory type 1 (Tr1) cells are characterized by their production of high levels of interleukin (IL)-10, which is crucial for their differentiation and suppressive function. We investigated the effects of IL-10 administered in combination with rapamycin on the induction of Tr1 cells that could mediate a state of tolerance in diabetic mice after pancreatic islet transplantation. The efficacy of this treatment was compared with IL-10 alone and standard immunosuppression. Stable long-term tolerance that was not reversible by alloantigen rechallenge was achieved only in mice treated with rapamycin plus IL 10. Tr1 cells that produced high levels of IL-10 and suppressed T-cell proliferation were isolated from splenocytes of rapamycin plus IL-10-treated mice after treatment withdrawal. In rapamycin plus IL-10-treated mice, endogenous IL 10 mediated an active state of tolerance, as was observed when the blockade of IL 10 activity rapidly induced graft rejection >100 days after transplantation. CD4(+) T-cells from rapamycin plus IL-10-treated mice transferred antigen specific tolerance in mice that received new transplants. Thus rapamycin plus IL 10 not only prevented allograft rejection but also induced Tr1 cells that mediated stable antigen-specific, long-term tolerance in vivo. PMID- 16380476 TI - Coexpression of CD25 and OX40 (CD134) receptors delineates autoreactive T-cells in type 1 diabetes. AB - T-cell-mediated loss of pancreatic beta-cells is the crucial event in the development of type 1 diabetes. The phenotypic characteristics of disease associated T-cells in type 1 diabetes have not yet been defined. The negative results from two intervention trials (the Diabetes Prevention Trial-Type 1 Diabetes and the European Nicotinamide Diabetes Intervention Trial) illustrate the need for technologies to specifically monitor ongoing autoimmune reactions. We used fluorescence-activated cell sorter analysis to study surface marker expression on T-cell lines specific for two major type 1 diabetes autoantigens, GAD65 and proinsulin. We then applied this knowledge in a cross-sectional approach to delineate the phenotype of circulating memory T-cells. The autoreactive T-cells of patients could be distinguished from those of control subjects by their coexpression of CD25 and CD134. Autoantigen-specific T-cells that recognized multiple GAD65- and preproinsulin-derived peptides and coexpressed CD25(+)CD134(+) were confined to patients (n = 32) and pre-diabetic probands (n = 5). Autoantigen-reactive T-cells in control subjects (n = 21) were CD25(+)CD134(-) and recognized fewer autoantigen-derived peptides. Insulin therapy did not induce CD25(+)CD134(+) T-cells in type 2 diabetic patients. The coexpression of CD25 and the costimulatory molecule CD134 on memory T-cells provides a novel marker for type 1 diabetes-associated T-cell immunity. The CD134 costimulatory molecule may also provide a novel therapeutic target in type 1 diabetes. PMID- 16380477 TI - A vHNF1/TCF2-HNF6 cascade regulates the transcription factor network that controls generation of pancreatic precursor cells. AB - Generation of pancreatic precursor cells in the endoderm is controlled by a network of transcription factors. Hepatocyte nuclear factor-6 (HNF6) is a key player in this network, because it controls the initiation of the expression of pancreatic and duodenal homeobox 1 (Pdx1), the earliest marker of pancreatic precursor cells. To further characterize this network, we have investigated how the expression of HNF6 is controlled in mouse endoderm, by using in vitro and in vivo protein-DNA interaction techniques combined with endoderm electroporation, transgenesis, and gene inactivation in embryos. We delineated Hnf6 regulatory regions that confer expression of a reporter gene in the embryonic endoderm but not in extraembryonic visceral endoderm. HNF6 expression in the embryonic endoderm was found to depend on an intronic enhancer. This enhancer contains functional binding sites for the tissue-specific factors of the forkhead box A and HNF1 families. Among the latter, variant HNF1 (vHNF1)/TCF2, which is expressed before HNF6 in the endoderm, was found to be critical for HNF6 expression. Therefore, the sequential activation of vHNF1, HNF6, and Pdx1 in the endoderm appears to control the generation of pancreatic precursors. This cascade may be used to benchmark in vitro differentiation of pancreatic precursor cells from embryonic stem cells, for cell therapy of diabetes. PMID- 16380478 TI - Evaluation of beta-cell replication in mice transgenic for hepatocyte growth factor and placental lactogen: comprehensive characterization of the G1/S regulatory proteins reveals unique involvement of p21cip. AB - We hypothesized that combined transgenic overexpression of hepatocyte growth factor (HGF) and placental lactogen in islets would lead to even greater increases in beta-cell mass and replication than either growth factor alone. This did not occur, suggesting that beta-cell replication is saturable or subject to molecular restraint. We therefore performed the first comprehensive G(1)/S cell cycle survey in islets, cataloguing the broad range of kinases, cyclins, and kinase inhibitors that control the G(1)/S transition in islets from normal, HGF, placental lactogen, and doubly transgenic mice. Many of the G(1)/S checkpoint regulators (E2Fs; pRb; p107; p130; cyclins D(1),(2),(3), A, and E; cdk-2; cdk-4; p15; p16; p18; p19; p21; p27; MDM2; p53; c-Myc; and Egr-1) are present in the murine islet. Most of these proteins were unaltered by overexpression of HGF or placental lactogen, either alone or in combination. In contrast, p21(cip) was uniquely, dramatically, and reproducibly upregulated in placental lactogen and HGF islets. p21(cip) was also present in, and upregulated in, proliferating human islets, localizing specifically in beta-cells and translocating to the nucleus on mitogenic stimulation. Homozygous p21(cip) loss releases islets from growth inhibition, markedly enhancing proliferation in response to HGF and placental lactogen. PMID- 16380479 TI - Glibenclamide treatment recruits beta-cell subpopulation into elevated and sustained basal insulin synthetic activity. AB - Use of sulfonylureas in diabetes treatment is based on their insulin-releasing effect on pancreatic beta-cells. Prolonged action is known to degranulate beta cells, but functional consequences have not been examined at the cellular level. This study investigates influences of in vivo (48-h) and in vitro (24-h) glibenclamide treatment on the functional state of the beta-cell population. Both conditions decreased cellular insulin content by >50% and caused an elevated basal insulin biosynthetic activity that was maintained for at least 24 h after drug removal. Glibenclamide stimulation of basal insulin synthesis was not achieved after a 2-h exposure; it required a calcium-dependent translational activity and involved an increase in the percent activated beta-cells (50% after glibenclamide pretreatment vs. 8% in control cells). The glibenclamide-activated beta-cell subpopulation corresponded to the degranulated beta-cell subpopulation that was isolated by fluorescence-activated cell sorter on the basis of lower cellular sideward scatter. Glibenclamide pretreatment did not alter cellular rates of glucose oxidation but sensitized beta-cells to glucose-induced changes in metabolic redox and insulin synthesis and release. In conclusion, chronic exposure to glibenclamide results in degranulation of a subpopulation of beta cells, which maintain an elevated protein and insulin synthetic activity irrespective of the presence of the drug and of glucose. Our study demonstrates that the in situ beta-cell population also exhibits a functional heterogeneity that can vary with drug treatment. Glibenclamide induces degranulated beta-cells with a sustained elevated basal activity that might increase the risk for hypoglycemic episodes. PMID- 16380480 TI - Effect of combined antisense oligonucleotides against high-glucose- and diabetes induced overexpression of extracellular matrix components and increased vascular permeability. AB - The effect of combined antisense oligonucleotides (AS-oligos) against overexpression of extracellular matrix (ECM) components, fibronectin, laminin, and collagen IV and on cell monolayer permeability was examined in rat microvascular endothelial cells (RMECs) grown in high glucose medium and on retinal vascular permeability in diabetic rats. RMECs grown in high glucose for 10 days and transfected with combined AS-oligos showed a significantly reduced fibronectin, laminin, and collagen IV protein level. In parallel studies, high glucose-induced excess monolayer permeability was also reduced in RMECs transfected with the combined AS-oligos. Similarly, diabetic rats intravitreally injected with the combined AS-oligos and examined after 2 months of diabetes showed significant reduction in retinal fibronectin, laminin, and collagen IV expression. In addition, vascular permeability, as determined from extravasation of fluorescein isothiocyanate-BSA in the surrounding areas of the retinal capillaries, was partially reduced in the combined AS-oligos-treated diabetic retinas. Our results indicate that the combined AS-oligos strategy is effective in simultaneously reducing fibronectin, collagen IV, and laminin overexpression and reducing vascular leakage in the retinal capillaries of diabetic rat retinas. The findings suggest that abnormal synthesis of ECM components may contribute to vascular leakage in the diabetic retina. PMID- 16380481 TI - Glucosamine-induced endoplasmic reticulum dysfunction is associated with accelerated atherosclerosis in a hyperglycemic mouse model. AB - Diabetes is a major independent risk factor for cardiovascular disease and stroke; however, the molecular and cellular mechanisms by which diabetes contributes to the development of vascular disease are not fully understood. Our previous studies demonstrated that endoplasmic reticulum (ER) stress-inducing agents, including homocysteine, promote lipid accumulation and activate inflammatory pathways-the hallmark features of atherosclerosis. We hypothesize that the accumulation of intracellular glucosamine observed in diabetes may also promote atherogenesis via a mechanism that involves ER stress. In support of this theory, we demonstrate that glucosamine can induce ER stress in cell types relevant to the development of atherosclerosis, including human aortic smooth muscle cells, monocytes, and hepatocytes. Furthermore, we show that glucosamine induced ER stress dysregulates lipid metabolism, leading to the accumulation of cholesterol in cultured cells. To examine the relevance of the ER stress pathway in vivo, we used a streptozotocin-induced hyperglycemic apolipoprotein E deficient mouse model of atherosclerosis. Using molecular biological and histological techniques, we show that hyperglycemia is associated with tissue specific ER stress, hepatic steatosis, and accelerated atherosclerosis. This novel mechanism may not only explain how diabetes and hyperglycemia promote atherosclerosis, but also provide a potential new target for therapeutic intervention. PMID- 16380482 TI - Nitric oxide cytoskeletal-induced alterations reverse the endothelial progenitor cell migratory defect associated with diabetes. AB - Stromal-derived factor-1 (SDF-1) is a critical chemokine for endothelial progenitor cell (EPC) recruitment to areas of ischemia, allowing these cells to participate in compensatory angiogenesis. The SDF-1 receptor, CXCR4, is expressed in developing blood vessels as well as on CD34+ EPCs. We describe that picomolar and nanomolar concentrations of SDF-1 differentially influence neovascularization, inducing CD34+ cell migration and EPC tube formation. CD34+ cells isolated from diabetic patients demonstrate a marked defect in migration to SDF-1. This defect is associated, in some but not all patients, with a cell surface activity of CD26/dipeptidyl peptidase IV, an enzyme that inactivates SDF 1. Diabetic CD34+ cells also do not migrate in response to vascular endothelial growth factor and are structurally rigid. However, incubating CD34+ cells with a nitric oxide (NO) donor corrects this migration defect and corrects the cell deformability. In addition, exogenous NO alters vasodilator-stimulated phosphoprotein and mammalian-enabled distribution in EPCs. These data support a common downstream cytoskeletal alteration in diabetic CD34+ cells that is independent of growth factor receptor activation and is correctable with exogenous NO. This inability of diabetic EPCs to respond to SDF-1 may contribute to aberrant tissue vascularization and endothelial repair in diabetic patients. PMID- 16380483 TI - The thromboxane receptor antagonist S18886 attenuates renal oxidant stress and proteinuria in diabetic apolipoprotein E-deficient mice. AB - Arachidonic acid metabolites, some of which may activate thromboxane A(2) receptors (TPr) and contribute to the development of diabetes complications, including nephropathy, are elevated in diabetes. This study determined the effect of blocking TPr with S18886 or inhibiting cyclooxygenase with aspirin on oxidative stress and the early stages of nephropathy in streptozotocin-induced diabetic apolipoprotein E(-/-) mice. Diabetic mice were treated with S18886 (5 mg . kg(-1) . day(-1)) or aspirin (30 mg . kg(-1) . day(-1)) for 6 weeks. Neither S18886 nor aspirin affected hyperglycemia or hypercholesterolemia. There was intense immunohistochemical staining for nitrotyrosine in diabetic mouse kidney. In addition, a decrease in manganese superoxide dismutase (MnSOD) activity was associated with an increase in MnSOD tyrosine-34 nitration. Tyrosine nitration was significantly reduced by S18886 but not by aspirin. Staining for the NADPH oxidase subunit p47(phox), inducible nitric oxide synthase, and 12-lipoxygenase was increased in diabetic mouse kidney, as were urine levels of 12 hydroxyeicosatetraenoic acid and 8-iso-prostaglandin F(2alpha). S18886 attenuated all of these markers of oxidant stress and inflammation. Furthermore, S18886 significantly attenuated microalbuminuria in diabetic mice and ameliorated histological evidence of diabetic nephropathy, including transforming growth factor-beta and extracellular matrix expression. Thus, in contrast to inhibiting cyclooxygenase, blockade of TPr may have therapeutic potential in diabetic nephropathy, in part by attenuating oxidative stress. PMID- 16380484 TI - Activation of AMP-activated protein kinase reduces hyperglycemia-induced mitochondrial reactive oxygen species production and promotes mitochondrial biogenesis in human umbilical vein endothelial cells. AB - We previously proposed that the production of hyperglycemia-induced mitochondrial reactive oxygen species (mtROS) is a key event in the development of diabetes complications. The association between the pathogenesis of diabetes and its complications and mitochondrial biogenesis has been recently reported. Because metformin has been reported to exert a possible additional benefit in preventing diabetes complications, we investigated the effect of metformin and 5 aminoimidazole-4-carboxamide ribonucleoside (AICAR) on mtROS production and mitochondrial biogenesis in cultured human umbilical vein endothelial cells. Treatment with metformin and AICAR inhibited hyperglycemia-induced intracellular and mtROS production, stimulated AMP-activated protein kinase (AMPK) activity, and increased the expression of peroxisome proliferator-activated response-gamma coactivator-1alpha (PGC-1alpha) and manganese superoxide dismutase (MnSOD) mRNAs. The dominant negative form of AMPKalpha1 diminished the effects of metformin and AICAR on these events, and an overexpression of PGC-1alpha completely blocked the hyperglycemia-induced mtROS production. In addition, metformin and AICAR increased the mRNA expression of nuclear respiratory factor-1 and mitochondrial DNA transcription factor A (mtTFA) and stimulated the mitochondrial proliferation. Dominant negative-AMPK also reduced the effects of metformin and AICAR on these observations. These results suggest that metformin normalizes hyperglycemia-induced mtROS production by induction of MnSOD and promotion of mitochondrial biogenesis through the activation of AMPK-PGC-1alpha pathway. PMID- 16380485 TI - High-density haplotype structure and association testing of the insulin-degrading enzyme (IDE) gene with type 2 diabetes in 4,206 people. AB - The insulin-degrading enzyme is responsible for the intracellular proteolysis of insulin. Its gene IDE is located on chromosome 10, in an area with suggestive linkage to type 2 diabetes and related phenotypes. Due to the impact of genetic variants of this gene in rodents and the function of its protein product, it has been proposed as a candidate gene for type 2 diabetes. Various groups have explored the role of the common genetic variation of IDE on insulin resistance and reported associations of various single nucleotide polymorphisms (SNPs) and haplotypes on both type 2 diabetes and glycemic traits. We sought to characterize the haplotype structure of IDE in detail and replicate the association of common variants with type 2 diabetes, fasting insulin, fasting glucose, and insulin resistance. We assessed linkage disequilibrium, selected single-marker and multimarker tags, and genotyped these markers in several case-control and family based samples totalling 4,206 Caucasian individuals. We observed no statistically significant evidence of association between single-marker or multimarker tests in IDE and type 2 diabetes. Nominally significant differences in quantitative traits are consistent with statistical noise. We conclude that common genetic variation at IDE is unlikely to confer clinically significant risk of type 2 diabetes in Caucasians. PMID- 16380486 TI - Increased lipid availability impairs insulin-stimulated ATP synthesis in human skeletal muscle. AB - Insulin resistance correlates with intramyocellular lipid content (IMCL) and plasma free fatty acids (FFAs) and was recently linked to mitochondrial dysfunction. We examined the underlying relationships by measuring skeletal muscle ATP synthase flux, glucose transport/phosphorylation, and IMCL in response to different plasma insulin and plasma FFA concentrations. Healthy men were studied twice during hyperinsulinemic-euglycemic clamps with (LIP) or without (CON) lipid infusion (plasma FFA: CON approximately 36 vs. LIP approximately 1,034 micromol/l, P < 0.001). ATP synthase flux, glucose-6-phosphate (G6P), and IMCL were determined before and during the clamp in calf muscle using (31)P and (1)H magnetic resonance spectroscopy. Plasma lipid elevation resulted in approximately 46% reduced whole-body glucose metabolism (180-360 min; P < 0.0001 vs. CON) and a 70% lower rise of G6P (P < 0.05 vs. CON) without significant changes in IMCL (LIP 117 +/- 12% vs. CON 93 +/- 3% of basal, P = 0.073). During the clamp, ATP synthase flux increased by approximately 60% under control conditions (P = 0.02 vs. baseline) and was 24% lower during lipid infusion (LIP 11.0 +/- 0.9 vs. CON 14.6 +/- 1.2 micromol . g muscle(-1) . min(-1), P < 0.05). Physiologically increased plasma FFA concentrations reduce insulin-stimulated muscle ATP synthase flux in parallel with induction of insulin resistance. PMID- 16380487 TI - Adiponectin does not cross the blood-brain barrier but modifies cytokine expression of brain endothelial cells. AB - Adiponectin has recently been reported to generate a negative energy balance by increasing energy expenditure. However, it is unclear whether such effects require the presence and direct action of the adiponectin protein in the central nervous system. In this study, neither radiolabeled nonglycosylated nor glycosylated globular adiponectin crossed the blood-brain barrier (BBB) in mice. In addition, adiponectin was not detectable in human cerebrospinal fluid using various established methods. Using murine cerebral microvessels, we demonstrated expression of adiponectin receptors, which are upregulated during fasting, in brain endothelium. Interestingly, treatment with adiponectin reduced secretion of the centrally active interleukin-6 from brain endothelial cells, a phenomenon that was paralleled by a similar trend of other proinflammatory cytokines. In summary, our data suggest that direct effects of endogenous adiponectin on central nervous system pathways are unlikely to exist. However, the identification of adiponectin receptors on brain endothelial cells and the finding of a modified secretion pattern of centrally active substances from BBB cells provides an alternate explanation as to how adiponectin may evoke effects on energy metabolism. PMID- 16380488 TI - Adipose triglyceride lipase: function, regulation by insulin, and comparison with adiponutrin. AB - Adipose triglyceride lipase (ATGL) is a recently described adipose-enriched protein with triglyceride-specific lipase activity. ATGL shares the greatest sequence homology with adiponutrin, a nutritionally regulated protein of unclear biological function. Here we present a functional analysis of ATGL and adiponutrin and describe their regulation by insulin. Retroviral-mediated overexpression of ATGL in 3T3-L1 adipocytes increased basal and isoproterenol stimulated glycerol and nonesterified fatty acid (NEFA) release, whereas siRNA mediated knockdown of ATGL had the opposite effect. In contrast, siRNA-mediated knockdown of adiponutrin in 3T3-L1 adipocytes had no effect on glycerol or NEFA release. In mice, both ATGL and adiponutrin are nutritionally regulated in adipose tissue, with ATGL being upregulated and adiponutrin being downregulated by fasting. In 3T3-L1 adipocytes, insulin decreased ATGL and increased adiponutrin expression in a dose- and time-dependent manner, suggesting that insulin directly mediates this nutritional regulation. In addition, adipose expression of ATGL was increased by insulin deficiency and decreased by insulin replacement in streptozotocin-induced diabetic mice and was increased in fat specific insulin receptor knockout mice, whereas adiponutrin showed the opposite pattern. These data suggest that murine ATGL but not adiponutrin contributes to net adipocyte lipolysis and that ATGL and adiponutrin are oppositely regulated by insulin both in vitro and in vivo. PMID- 16380489 TI - Interleukin-7 is a survival factor for CD4+ CD25+ T-cells and is expressed by diabetes-suppressive dendritic cells. AB - Dendritic cells can facilitate allograft survival and prevent autoimmunity via direct and indirect cell-mediated mechanisms. Recent studies demonstrate that immunoregulatory dendritic cells (iDCs) confer immune hyporesponsiveness in part through CD4(+) CD25(+) T regulatory cells (Tregs). Herein, we provide evidence to support the hypothesis that dendritic cells derived from NOD mice and engineered ex vivo to exhibit suppressed expression of the CD40, CD80, and CD86 costimulatory molecules motivate an increase in the prevalence of regulatory CD4(+) CD25(+) T-cells via interleukin (IL)-7. Unlike control dendritic cells, these dendritic cells expressed significant levels of IL-7. Exogenous addition of IL-7 to NOD T-cells did not promote expansion or proliferation, but instead selectively maintained the number of CD4(+) CD25(+) T-cells by inhibiting activation of apoptosis in these cells. In vitro, IL-7 receptor alpha-chain (IL 7Ralpha) was expressed at significantly higher levels on CD4(+) CD25(+) T-cells compared with CD4(+) CD25(-) T-cells irrespective of resting or stimulated state. In vivo, CD4(+) CD25(+) T-cells obtained from NOD-scid mice reconstituted with ex vivo engineered iDCs and NOD splenocytes expressed significantly higher levels of IL-7Ralpha compared with levels in the CD4(+) CD25(-) subset, especially in diabetes-suppressive dendritic cell-administered NOD-scid recipients. Taken together, our data suggest a novel mechanism by which iDCs delay autoimmunity through the CD4(+) CD25(+) Treg pathway and suggest IL-7 as a survival factor for these putative Tregs, which express the alpha-chain of its receptor at considerably higher levels than CD4(+) CD25(-) T-cells. PMID- 16380490 TI - Novel leptin receptor mutation in NOD/LtJ mice suppresses type 1 diabetes progression: II. Immunologic analysis. AB - Recently, we identified in normally type 1 diabetes-prone NOD/LtJ mice a spontaneous new leptin receptor (LEPR) mutation (designated Lepr(db-5J)) producing juvenile obesity, hyperglycemia, hyperinsulinemia, and hyperleptinemia. This early type 2 diabetes syndrome suppressed intra-islet insulitis and permitted spontaneous diabetes remission. No significant differences in plasma corticosterone, splenic CD4(+) or CD8(+) T-cell percentages, or functions of CD3(+) T-cells in vitro distinguished NOD wild-type from mutant mice. Yet splenocytes from hyperglycemic mutant donors failed to transfer type 1 diabetes into NOD.Rag1(-/-) recipients over a 13-week period, whereas wild-type donor cells did so. This correlated with significantly reduced (P < 0.01) frequencies of insulin and islet-specific glucose-6-phosphatase catalytic subunit-related protein-reactive CD8(+) T-effector clonotypes in mutant mice. Intra-islet insulitis was also significantly suppressed in lethally irradiated NOD-Lepr(db 5J)/Lt recipients reconstituted with wild-type bone marrow (P < 0.001). In contrast, type 1 diabetes eventually developed when mutant marrow was transplanted into irradiated wild-type recipients. Mitogen-induced T-cell blastogenesis was significantly suppressed when splenic T-cells from both NOD/Lt and NOD-Lepr(db-5J)/Lt donors were incubated with irradiated mutant peritoneal exudate cells (P < 0.005). In conclusion, metabolic disturbances elicited by a type 2 diabetes syndrome (insulin and/or leptin resistance, but not hypercorticism) appear to suppress type 1 diabetes development in NOD-Lepr(db 5J)/Lt by inhibiting activation of T-effector cells. PMID- 16380491 TI - Transforming growth factor-beta and natural killer T-cells are involved in the protective effect of a bacterial extract on type 1 diabetes. AB - The onset of type 1 diabetes in NOD mice is delayed by oral administration of a bacterial extract (OM-85) and can be completely prevented by its intraperitoneal administration. Optimal prevention is observed when starting treatment at 3 or 6 weeks of age, and some effect is still observed with treatment at 10 weeks of age. Using genetically deficient mice and cytokine-neutralizing monoclonal antibodies, we demonstrate here that the therapeutic effect does not involve T helper type 2 cytokines (interleukin [IL]-4 and -10) but is tightly dependent on transforming growth factor (TGF)-beta. Natural killer T-cells also participate in the therapeutic effect because CD1d(-/-) NOD mice are partially resistant to the protective effect of OM-85. The question remains of the specificity of the protective effect of OM-85, which may include proinflammatory components. It will thus be important to further characterize the molecular components that afford protection from type 1 diabetes. Lipopolysaccharide is excluded, but other Toll like receptor (TLR) agonists could be involved because OM-85 stimulated dendritic cells and induced TGF-beta production by splenocytes in a TLR-2-, TLR-4-, and MyD88-dependent fashion. PMID- 16380492 TI - The diabetes type 1 locus Idd6 modulates activity of CD4+CD25+ regulatory T cells. AB - The genetic locus Idd6 confers susceptibility to the spontaneous development of type 1 diabetes in the NOD mouse. Our studies on disease resistance of the congenic mouse strain NOD.C3H 6.VIII showed that Idd6 influences T-cell activities in the peripheral immune system and suggest that a major mechanism by which the Idd6 locus modifies diabetes development is via modulation of regulatory T-cell activities. Our transfer experiments using total splenocytes and purified T-cells demonstrated that the locus specifically controls the efficiency of disease protection mediated by the regulatory CD4(+)CD25(+) T-cell subset. Our data also implicate the Idd6 locus in controlling the balance between infiltrating lymphocytes and antigen-presenting cells within the pancreatic islet. PMID- 16380493 TI - Leucine regulation of glucokinase and ATP synthase sensitizes glucose-induced insulin secretion in pancreatic beta-cells. AB - We have recently shown that leucine culture upregulates ATP synthase beta-subunit (ATPSbeta) and increases ATP level, cytosolic Ca(2+), and glucose-induced insulin secretion in rat islets. The aim is to test whether glucokinase expression is also affected in rat islets and its role in glucose sensitization during leucine culture. Leucine culture increased glucose-induced NAD(P)H level at 1 and 2 days but not at 1 week. The half-maximal effective concentration of the glucose response curve for NAD(P)H was left-shifted from 5-7 to 2-3 mmol/l. The effect was dose dependent and rapamycin insensitive. Leucine culture did not affect glyceraldehyde effects on NAD(P)H. Leucine pretreatment for 30 min had no effects on NAD(P)H levels. Leucine culture for 2 days also increased glucose-induced cytosolic Ca(2+) elevation, ATP level, and insulin secretion. Leucine increase of glucokinase mRNA levels occurred as early as day 1 and lasted through 1 week. That of ATPSbeta did not occur until day 2 and lasted through 1 week. Leucine effects on both mRNAs were dose dependent. The upregulation of both genes was confirmed by Western blotting. Leucine culture also increased glucose-induced insulin secretion, ATP level, glucokinase, and ATPSbeta levels of type 2 diabetic human islets. In conclusion, leucine culture upregulates glucokinase, which increases NAD(P)H level, and ATPSbeta, which increases oxidation of NADH and production of ATP. The combined upregulation of both genes increases glucose induced cytosolic Ca(2+) and insulin secretion. PMID- 16380494 TI - Effects of hyperglycemia and hyperinsulinemia on circulating tissue factor procoagulant activity and platelet CD40 ligand. AB - Individuals with chronically elevated glucose and/or insulin levels, i.e., most patients with type 2 diabetes, have accelerated atherosclerosis and are prone to acute vascular events. We have tested the hypothesis that hyperglycemia and/or hyperinsulinemia singly or combined may increase tissue factor, the primary initiator of blood coagulation. We have determined changes in circulating tissue factor procoagulant activity (PCA) and other procoagulation proteins in healthy volunteers exposed to 24 h of selective hyperinsulinemia, selective hyperglycemia, or combined hyperinsulinemia and hyperglycemia. Combined elevations of plasma insulin and glucose levels for 24 h produced a ninefold increase in tissue factor PCA, which was associated with an increase in monocyte tissue factor protein (flow cytometry) and mRNA (RT-PCR), increases in plasma thrombin-antithrombin complexes, prothrombin fragment 1.2, factor VIII coagulant activity, and platelet CD40 ligand as well as decreases in factor VIIa, factor VII coagulant activities, and factor VII antigen. Effects of selective hyperinsulinemia and selective hyperglycemia were less striking but appeared to be additive. We conclude that hyperinsulinemia and hyperglycemia but particularly the combination of both create a prothrombotic state and in addition may be proinflammatory and proatherogenic because of the proinflammatory actions of CD40 ligand and tissue factor. PMID- 16380495 TI - Phagocytic NADPH oxidase overactivity underlies oxidative stress in metabolic syndrome. AB - Oxidative stress plays a critical role in the pathogenesis of atherosclerosis in patients with metabolic syndrome. This study aimed to investigate whether a relationship exists between phagocytic NADPH oxidase activity and oxidative stress and atherosclerosis in metabolic syndrome patients. The study was performed in 56 metabolic syndrome patients (metabolic syndrome group), 99 patients with one or two cardiovascular risk factors (cardiovascular risk factor group), and 28 healthy subjects (control group). NADPH oxidase expression and activity was augmented (P < 0.05) in metabolic syndrome compared with cardiovascular risk factor and control groups. Insulin was enhanced (P < 0.05) in metabolic syndrome patients compared with cardiovascular risk factor and control groups and correlated with NADPH oxidase activity in the overall population. Insulin stimulated NADPH oxidase activity; this effect was abolished by a specific protein kinase C inhibitor. Oxidized LDL and nitrotyrosine levels and carotid intima-media thickness were increased (P < 0.05) in the metabolic syndrome group compared with cardiovascular risk factor and control groups and correlated with NADPH oxidase activity in the overall population. These findings suggest that phagocytic NADPH oxidase overactivity is involved in oxidative stress and atherosclerosis in metabolic syndrome patients. Our findings also suggest that hyperinsulinemia may contribute to oxidative stress in metabolic syndrome patients through activation of NADPH oxidase. PMID- 16380496 TI - Natural antibiotics and insulin sensitivity: the role of bactericidal/permeability-increasing protein. AB - The innate immune system can immediately respond to microorganism intrusion by helping to prevent further invasion. Bactericidal/permeability-increasing protein (BPI) is a major constituent of neutrophils that possesses anti-inflammatory properties. Inflammation is increasingly recognized as a component of the metabolic syndrome. We hypothesized that the production of BPI could be linked to insulin sensitivity and glucose tolerance. We studied circulating BPI across categories of glucose tolerance. We also studied whether these cross-sectional associations were of functional importance. For this reason, we investigated circulating bioactive lipopolysaccharide and the effects of changing insulin action-after treatment with an insulin sensitizer (metformin)-on circulating BPI in subjects with glucose intolerance. Finally, we tested whether a 3' untranslated region (UTR) BPI polymorphism led to differences in BPI and insulin action among nondiabetic subjects. Age- and BMI-adjusted circulating BPI was significantly lower among patients with type 2 diabetes. Circulating BPI correlated negatively with fasting and postload glucose and insulin concentrations. In subjects with glucose intolerance, BPI was also linked to BMI, waist-to-hip ratio, and age- and BMI-adjusted insulin sensitivity. Bioactive lipopolysaccharide was negatively correlated with circulating BPI (r = -0.57, P < 0.0001) and positively with plasma lipopolysaccharide-binding protein (r = 0.54, P = 0.002). In parallel to improved insulin sensitivity, plasma BPI significantly increased in the metformin group but not in the placebo group. A 3'-UTR BPI polymorphism was simultaneously associated with plasma BPI concentration, waist to-hip ratio, fasting and postload insulin concentration, fasting plasma triglycerides, and insulin sensitivity. These findings suggest that this component of the innate immune system is associated with metabolic pathways. PMID- 16380497 TI - Glucose-induced reactive oxygen species cause apoptosis of podocytes and podocyte depletion at the onset of diabetic nephropathy. AB - Diabetic nephropathy is the most common cause of end-stage renal disease in the U.S. Recent studies demonstrate that loss of podocytes is an early feature of diabetic nephropathy that predicts its progressive course. Cause and consequences of podocyte loss during early diabetic nephropathy remain poorly understood. Here, we demonstrate that podocyte apoptosis increased sharply with onset of hyperglycemia in Ins2(Akita) (Akita) mice with type 1 diabetes and Lepr(db/db) (db/db) mice with obesity and type 2 diabetes. Podocyte apoptosis coincided with the onset of urinary albumin excretion (UAE) and preceded significant losses of podocytes in Akita (37% reduction) and db/db (27% reduction) mice. Increased extracellular glucose (30 mmol/l) rapidly stimulated generation of intracellular reactive oxygen species (ROS) through NADPH oxidase and mitochondrial pathways and led to activation of proapoptotic p38 mitogen-activated protein kinase and caspase 3 and to apoptosis of conditionally immortalized podocytes in vitro. Chronic inhibition of NADPH oxidase prevented podocyte apoptosis and ameliorated podocyte depletion, UAE, and mesangial matrix expansion in db/db mice. In conclusion, our results demonstrate for the first time that glucose-induced ROS production initiates podocyte apoptosis and podocyte depletion in vitro and in vivo and suggest that podocyte apoptosis/depletion represents a novel early pathomechanism(s) leading to diabetic nephropathy in murine type 1 and type 2 diabetic models. PMID- 16380498 TI - Increased renal arterial resistance predicts the course of renal function in type 2 diabetes with microalbuminuria. AB - Type 2 diabetic patients often die because of end-stage renal failure, but no definitive reliable factor predicting long-term renal outcome has been identified. We tested whether a renal arterial resistance index (R/I) > or =80, using Doppler ultrasound technique, was predictive of worsening renal function. The primary end points of the study were 1) the course of glomerular filtration rate (GFR) and 2) the albumin excretion rate in 157 microalbuminuric, hypertensive, type 2 diabetic patients after a 7.8-year follow-up period (range 7.1-9.2). Kaplan-Meier curves for the primary end point (decrease of GFR > or = 3.0 ml/min per 1.73 m(2) per year) was two to three times more frequently observed in patients with R/I > or =80. Four- to fivefold fewer patients showed a regression to normoalbuminuria during the follow-up period from baseline microalbuminuria in the cohort with R/I > or =80. Overt proteinuria did develop in 24% of patients with R/I > or =80 and in 5% of patients with R/I <80 (P < 0.01). In conclusion, intrarenal arterial resistance appears to play a nontrivial role in deteriorating renal function in type 2 diabetic patients. R/I is a noninvasive diagnostic procedure, which strongly predicts the outcome of renal function in type 2 diabetic patients, even when GFR patterns are still normal. PMID- 16380500 TI - Adiponectin multimeric complexes and the metabolic syndrome trait cluster. AB - Adiponectin circulates in human plasma mainly as a 180-kDa low molecular weight (LMW) hexamer and a high molecular weight (HMW) multimer of approximately 360 kDa. We comprehensively examined the relationships between circulating levels of total adiponectin, adiponectin multimers, and the relative distribution (i.e., ratio) of multimeric forms with key features of the metabolic syndrome. Total adiponectin (r = 0.45), HMW (r = 0.47), LMW (r = 0.31), and HMW-to-total adiponectin ratio (r = 0.29) were significantly correlated with insulin stimulated glucose disposal rate. Similarly, total (r = -0.30), HMW (r = -0.38), and HMW-to-total adiponectin ratio (r = -0.34) were correlated with central fat distribution but not with total fat mass or BMI. Regarding energy metabolism, although there were no effects on resting metabolic rate, total (r = 0.41) and HMW (r = 0.44) were associated with increasing rates of fat oxidation. HMW-to total adiponectin ratio increased as a function of total adiponectin, and it was HMW quantity (not total or HMW-to-total adiponectin ratio or LMW) that was primarily responsible for all of these relationships. Impact on nuclear magnetic resonance lipoprotein subclasses was assessed. HMW and total adiponectin were correlated with decreases in large VLDL (r = -0.44 and -0.41); decreases in small LDL (r = -0.41 and -0.36) and increases in large LDL (r = 0.36 and 0.30) particle concentrations accompanied by increased LDL particle size (r = 0.47 and 0.39); and increases in large HDL (r = 0.45 and 0.37) and HDL particle size (r = 0.53 and 0.47). Most of these correlations persisted after adjustment for metabolic covariables. In conclusion, first, serum adiponectin is associated with increased insulin sensitivity, reduced abdominal fat, and high basal lipid oxidation; however, it is HMW quantity, not total or HMW-to-total adiponectin ratio, that is primarily responsible for these relationships. Second, reduced quantities of HMW independently recapitulate the lipoprotein subclass profile associated with insulin resistance after correcting for glucose disposal rate and BMI. Finally, HMW adiponectin is an important factor in explaining the metabolic syndrome. PMID- 16380499 TI - Muraglitazar, a novel dual (alpha/gamma) peroxisome proliferator-activated receptor activator, improves diabetes and other metabolic abnormalities and preserves beta-cell function in db/db mice. AB - Muraglitazar, a novel dual (alpha/gamma) peroxisome proliferator-activated receptor (PPAR) activator, was investigated for its antidiabetic properties and its effects on metabolic abnormalities in genetically obese diabetic db/db mice. In db/db mice and normal mice, muraglitazar treatment modulates the expression of PPAR target genes in white adipose tissue and liver. In young hyperglycemic db/db mice, muraglitazar treatment (0.03-50 mg . kg(-1) . day(-1) for 2 weeks) results in dose-dependent reductions of glucose, insulin, triglycerides, free fatty acids, and cholesterol. In older hyperglycemic db/db mice, longer-term muraglitazar treatment (30 mg . kg(-1) . day(-1) for 4 weeks) prevents time dependent deterioration of glycemic control and development of insulin deficiency. In severely hyperglycemic db/db mice, muraglitazar treatment (10 mg . kg(-1) . day(-1) for 2 weeks) improves oral glucose tolerance and reduces plasma glucose and insulin levels. In addition, treatment increases insulin content in the pancreas. Finally, muraglitazar treatment increases abnormally low plasma adiponectin levels, increases high-molecular weight adiponectin complex levels, reduces elevated plasma corticosterone levels, and lowers elevated liver lipid content in db/db mice. The overall conclusions are that in db/db mice, the novel dual (alpha/gamma) PPAR activator muraglitazar 1) exerts potent and efficacious antidiabetic effects, 2) preserves pancreatic insulin content, and 3) improves metabolic abnormalities such as hyperlipidemia, fatty liver, low adiponectin levels, and elevated corticosterone levels. PMID- 16380501 TI - Variants in the human insulin gene that affect pre-mRNA splicing: is -23HphI a functional single nucleotide polymorphism at IDDM2? AB - Predisposition to type 1 diabetes and juvenile obesity is influenced by the susceptibility locus IDDM2 that includes the insulin gene (INS). Although the risk conferred by IDDM2 has been attributed to a minisatellite upstream of INS, intragenic variants have not been ruled out. We examined whether INS polymorphisms affect pre-mRNA splicing and proinsulin secretion using minigene reporter assays. We show that IVS1-6A/T (-23HphI+/-) is a key INS variant that influences alternative splicing of intron 1 through differential recognition of its 3' splice site. The A allele resulted in an increased production of mature transcripts with a long 5' leader in several cell lines, and the extended mRNAs generated more proinsulin in culture supernatants than natural transcripts. The longer mRNAs were significantly overrepresented among beta-cell-expressed sequenced tags containing the A allele as compared with those with T alleles. In addition, we show that a rare insertion/deletion polymorphism IVS1+5insTTGC (IVS 69), which is exclusively present in Africans, activated a downstream cryptic 5' splice site, extending the 5' leader by 30 bp. These results indicate that 23HphI and IVS-69 are the most important INS variants affecting pre-mRNA splicing and suggest that -23HphI+/- is a common functional single nucleotide polymorphism at IDDM2. PMID- 16380502 TI - The emerging role of the COP9 signalosome in cancer. AB - In the last several years, multiple lines of evidence have suggested that the COP9 signalosome (CSN) plays a significant role in the regulation of multiple cancers and could be an attractive target for therapeutic intervention. First, the CSN plays a key role in the regulation of Cullin-containing ubiquitin E3 ligases that are central mediators of a variety of cellular functions essential during cancer progression. Second, several studies suggest that the individual subunits of the CSN, particularly CSN5, might regulate oncogenic and tumor suppressive functions independently of, or coordinately with, the CSN holocomplex. Thus, deregulation of CSN subunit function can have a dramatic effect on diverse cellular functions, including the maintenance of DNA fidelity, cell cycle control, DNA repair, angiogenesis, and microenvironmental homeostasis that are critical for tumor development. Additionally, clinical studies have suggested that the expression or localization of some CSN subunits correlate to disease progression or clinical outcome in a variety of tumor types. Although the study of CSN function in relation to tumor progression is in its infancy, this review will address current studies in relation to cancer initiation, progression, and potential for therapeutic intervention. PMID- 16380503 TI - Comprehensive profiling of 8p11-12 amplification in breast cancer. AB - In human carcinomas, especially breast cancer, chromosome arm 8p is frequently involved in complex chromosomal rearrangements that combine amplification at 8p11 12, break in the 8p12-21 region, and loss of 8p21-ter. Several studies have identified putative oncogenes in the 8p11-12 amplicon. However, discrepancies and the lack of knowledge on the structure of this amplification lead us to think that the actual identity of the oncogenes is not definitively established. We present here a comprehensive study combining genomic, expression, and chromosome break analyses of the 8p11-12 region in breast cell lines and primary breast tumors. We show the existence of four amplicons at 8p11-12 using array comparative genomic hybridization. Gene expression analysis of 123 samples using DNA microarrays identified 14 genes significantly overexpressed in relation to amplification. Using fluorescence in situ hybridization analysis on tissue microarrays, we show the existence of a cluster of breakpoints spanning a region just telomeric to and associated with the amplification. Finally, we show that 8p11-12 amplification has a pejorative effect on survival in breast cancer. PMID- 16380504 TI - Identification of mitogen-activated protein kinase signaling pathways that confer resistance to endoplasmic reticulum stress in Saccharomyces cerevisiae. AB - Hypoxia activates all components of the unfolded protein response (UPR), a stress response initiated by the accumulation of unfolded proteins within the endoplasmic reticulum (ER). Our group and others have shown previously that the UPR, a hypoxia-inducible factor-independent signaling pathway, mediates cell survival during hypoxia and is required for tumor growth. Identifying new genes and pathways that are important for survival during ER stress may lead to the discovery of new targets in cancer therapy. Using the set of 4,728 homozygous diploid deletion mutants in budding yeast, Saccharomyces cerevisiae, we did a functional screen for genes that conferred resistance to ER stress-inducing agents. Deletion mutants in 56 genes showed increased sensitivity under ER stress conditions. Besides the classic UPR pathway and genes related to calcium homeostasis, we report that two additional pathways, including the SLT2 mitogen activated protein kinase (MAPK) pathway and the osmosensing MAPK pathway, were also required for survival during ER stress. We further show that the SLT2 MAPK pathway was activated during ER stress, was responsible for increased resistance to ER stress, and functioned independently of the classic IRE1/HAC1 pathway. We propose that the SLT2 MAPK pathway is an important cell survival signaling pathway during ER stress. This study shows the feasibility of using the yeast deletion pool to identify relevant mammalian orthologues of the UPR. PMID- 16380505 TI - Bone morphogenetic protein-2-induced transformation involves the activation of mammalian target of rapamycin. AB - Bone morphogenetic protein-2 (BMP-2) is an evolutionary conserved protein that is essential for embryonic development. BMP-2 is highly expressed in approximately 98% of human lung carcinomas with little expression in normal lung tissues. BMP-2 has been shown to enhance mobility, invasiveness, and metastasis of cancer cell lines. During development, BMP-2 induces the proto-oncogene phosphoinositide 3 kinase (PI3K)/mammalian target of rapamycin (mTOR) signaling pathway to regulate stem cell differentiation. We show that BMP-2 induces the phosphorylation of mTOR in A549 and H1299 lung cancer cell lines, which is attenuated by the PI3K antagonists LY-294002 and wortmannin. p70S6 kinase, which is a direct downstream target of mTOR, is also regulated by BMP-2 in lung cancer cell lines. We find that BMP-2 induces cyclin E in A549 and H1299 cells, which is mediated by the PI3K/mTOR signaling pathway. The regulation of cyclin E by BMP-2 occurs through a Smad 1/5-independent mechanism. Forced expression of BMP-2 in A549 cells (A549/BMP-2) induces transformation as shown by an increase in foci formation. The mTOR antagonist, rapamycin, prevented foci formation of the A549/BMP-2 cells. This study provides evidence that BMP-2-mediated transformation of lung cancer cells involves the activation of the PI3K/mTOR signaling pathway. PMID- 16380506 TI - Interleukin-12 up-regulates Fas expression in human osteosarcoma and Ewing's sarcoma cells by enhancing its promoter activity. AB - Interleukin-12 (IL-12) has shown significant antitumor activity in several preclinical animal tumor models. Our previous studies showed that IL-12 inhibited tumor growth in human osteosarcoma and Ewing's sarcoma animal model. Decreased Fas expression in osteosarcoma increased the lung metastatic potential. In this study, we further examined the mechanism of IL-12 antitumor activity and showed that IL-12 significantly increased Fas expression in both human osteosarcoma cells LM7 and Ewing's sarcoma cells TC71. Up-regulation of Fas expression increased their sensitivity to Fas-induced cell apoptosis. Constructs of the Fas promoter linked to a luciferase reporter gene were used to determine the promoter activity. IL-12 increased Fas promoter activity 4.2- and 4.9-fold in TC71 and LM7 cells, respectively. Time course studies have shown that recombinant IL-12 stimulated Fas promoter activity at 2 hours, reached the peak level at 4 hours, and then declined at 24 hours. To investigate whether IL-12 specifically enhanced Fas promoter activity, we determined whether another gene (E1A) was able to stimulate Fas promoter activity. We also evaluated effect of IL-12 on the topoisomerase IIalpha promoter. The results indicated that E1A but not IL-12 stimulated topoisomerase IIalpha promoter activity. E1A failed to increase Fas promoter activity. We also found that kappaB-Sp1 element at position -295 to -286 in Fas promoter was essential for IL-12-induced activation, and nuclear factor kappaB transcription factor was activated after IL-12 treatment in TC71 cells. These results indicate that IL-12 up-regulates Fas expression in human osteosarcoma and Ewing's sarcoma by enhancing Fas promoter activity. Understanding this mechanism may lead to new therapeutic approaches for the treatment of sarcoma involving the use of IL-12. PMID- 16380507 TI - Histone deacetylase 2-mediated deacetylation of the glucocorticoid receptor enables NF-kappaB suppression. AB - Glucocorticoids are the most effective antiinflammatory agents for the treatment of chronic inflammatory diseases even though some diseases, such as chronic obstructive pulmonary disease (COPD), are relatively glucocorticoid insensitive. However, the molecular mechanism of this glucocorticoid insensitivity remains uncertain. We show that a defect of glucocorticoid receptor (GR) deacetylation caused by impaired histone deacetylase (HDAC) 2 induces glucocorticoid insensitivity toward nuclear factor (NF)-kappaB-mediated gene expression. Specific knockdown of HDAC2 by RNA interference resulted in reduced sensitivity to dexamethasone suppression of interleukin 1beta-induced granulocyte/macrophage colony-stimulating factor production. Loss of HDAC2 did not reduce GR nuclear translocation, GR binding to glucocorticoid response element (GRE) on DNA, or GR induced DNA or gene induction but inhibited the association between GR and NF kappaB. GR becomes acetylated after ligand binding, and HDAC2-mediated GR deacetylation enables GR binding to the NF-kappaB complex. Site-directed mutagenesis of K494 and K495 reduced GR acetylation, and the ability to repress NF-kappaB-dependent gene expression becomes insensitive to histone deacetylase inhibition. In conclusion, we show that overexpression of HDAC2 in glucocorticoid insensitive alveolar macrophages from patients with COPD is able to restore glucocorticoid sensitivity. Thus, reduction of HDAC2 plays a critical role in glucocorticoid insensitivity in repressing NF-kappaB-mediated, but not GRE mediated, gene expression. PMID- 16380508 TI - LAT-mediated signaling in CD4+CD25+ regulatory T cell development. AB - Engagement of the T cell receptor for antigen (TCR) induces formation of signaling complexes mediated through the transmembrane adaptor protein, the linker for activation of T cells (LAT). LAT plays an important role in T cell development, activation, and homeostasis. A knock-in mutation at Tyr136, which is the phospholipase C (PLC)-gamma1-binding site in LAT, leads to a severe autoimmune disease in mice. In this study, we show that CD4+CD25+ T reg cells that expressed Foxp3 transcription factor were nearly absent in both thymus and peripheral lymphoid organs of LAT(Y136F) mice. This defect was not a result of the autoimmune environment as LAT(Y136F) T reg cells also failed to develop in healthy LAT-/- mice that received mixed wild-type and LAT(Y136F) bone marrow cells. Moreover, adoptive transfer of normal CD4+CD25+ T reg cells protected neonatal LAT(Y136F) mice from developing this disease. These T reg cells effectively controlled expansion of CD4+ T cells in LAT(Y136F) mice likely via granzymes and/or TGF-beta-mediated suppression. Furthermore, ectopic expression of Foxp3 conferred a suppressive function in LAT(Y136F) T cells. Our data indicate that the LAT-PLC-gamma1 interaction plays a critical role in Foxp3 expression and the development of CD4+CD25+ T reg cells. PMID- 16380509 TI - An essential role for complement C5a in the pathogenesis of septic cardiac dysfunction. AB - Defective cardiac function during sepsis has been referred to as "cardiomyopathy of sepsis." It is known that sepsis leads to intensive activation of the complement system. In the current study, cardiac function and cardiomyocyte contractility have been evaluated in rats after cecal ligation and puncture (CLP). Significant reductions in left ventricular pressures occurred in vivo and in cardiomyocyte contractility in vitro. These defects were prevented in CLP rats given blocking antibody to C5a. Both mRNA and protein for the C5a receptor (C5aR) were constitutively expressed on cardiomyocytes; both increased as a function of time after CLP. In vitro addition of recombinant rat C5a induced dramatic contractile dysfunction in both sham and CLP cardiomyocytes, but to a consistently greater degree in cells from CLP animals. These data suggest that CLP induces C5aR on cardiomyocytes and that in vivo generation of C5a causes C5a C5aR interaction, causing dysfunction of cardiomyocytes, resulting in compromise of cardiac performance. PMID- 16380510 TI - Regulation of the germinal center gene program by interferon (IFN) regulatory factor 8/IFN consensus sequence-binding protein. AB - Interferon (IFN) consensus sequence-binding protein/IFN regulatory factor 8 (IRF8) is a transcription factor that regulates the differentiation and function of macrophages, granulocytes, and dendritic cells through activation or repression of target genes. Although IRF8 is also expressed in lymphocytes, its roles in B cell and T cell maturation or function are ill defined, and few transcriptional targets are known. Gene expression profiling of human tonsillar B cells and mouse B cell lymphomas showed that IRF8 transcripts were expressed at highest levels in centroblasts, either from secondary lymphoid tissue or transformed cells. In addition, staining for IRF8 was most intense in tonsillar germinal center (GC) dark-zone centroblasts. To discover B cell genes regulated by IRF8, we transfected purified primary tonsillar B cells with enhanced green fluorescent protein-tagged IRF8, generated small interfering RNA knockdowns of IRF8 expression in a mouse B cell lymphoma cell line, and examined the effects of a null mutation of IRF8 on B cells. Each approach identified activation-induced cytidine deaminase (AICDA) and BCL6 as targets of transcriptional activation. Chromatin immunoprecipitation studies demonstrated in vivo occupancy of 5' sequences of both genes by IRF8 protein. These results suggest previously unappreciated roles for IRF8 in the transcriptional regulation of B cell GC reactions that include direct regulation of AICDA and BCL6. PMID- 16380511 TI - Purified hematopoietic stem cell engraftment of rare niches corrects severe lymphoid deficiencies without host conditioning. AB - In the absence of irradiation or other cytoreductive conditioning, endogenous hematopoietic stem cells (HSCs) are thought to fill the unique niches within the bone marrow that allow maintenance of full hematopoietic potential and thus prevent productive engraftment of transplanted donor HSCs. By transplantation of purified exogenous HSCs into unconditioned congenic histocompatible strains of mice, we show that approximately 0.1-1.0% of these HSC niches are available for engraftment at any given point and find no evidence that endogenous HSCs can be displaced from the niches they occupy. We demonstrate that productive engraftment of HSCs within these empty niches is inhibited by host CD4+ T cells that recognize very subtle minor histocompatibility differences. Strikingly, transplantation of purified HSCs into a panel of severe combined immunodeficient (SCID) mice leads to a rapid and complete rescue of lymphoid deficiencies through engraftment of these very rare niches and expansion of donor lymphoid progenitors. We further demonstrate that transient antibody-mediated depletion of CD4+ T cells allows short-term HSC engraftment and regeneration of B cells in a mouse model of B(-) non-SCID. These experiments provide a general mechanism by which transplanted HSCs can correct hematopoietic deficiencies without any host conditioning or with only highly specific and transient lymphoablation. PMID- 16380512 TI - Dual specificity phosphatase 1 (DUSP1) regulates a subset of LPS-induced genes and protects mice from lethal endotoxin shock. AB - Activation of the mitogen-activated protein kinase (MAPK) cascade after Toll-like receptor stimulation enables innate immune cells to rapidly activate cytokine gene expression. A balanced response to signals of infectious danger requires that cellular activation is transient. Here, we identify the MAPK phosphatase dual specificity phosphatase 1 (DUSP1) as an essential endogenous regulator of the inflammatory response to lipopolysaccharide (LPS). DUSP1-deficient (DUSP1-/-) bone marrow-derived macrophages showed selectively prolonged activation of p38 MAPK and increased cytokine production. Intraperitoneal challenge of DUSP1-/- mice with LPS caused increased lethality and overshooting production of interleukin (IL)-6 and tumor necrosis factor alpha. Transcriptional profiling revealed that DUSP1 controls a significant fraction of LPS-induced genes, which includes IL-6 and IL-10 as well as the chemokines CCL3, CCL4, and CXCL2. In contrast, the expression of the important mediators of endotoxin lethality, interferon gamma and IL-12, was not significantly altered by the absence of DUSP1. These data together demonstrate a specific regulatory role of DUSP1 in controlling a subset of LPS-induced genes that determines the outcome of endotoxin shock. PMID- 16380514 TI - Targeting brain AT1 receptors by RNA interference. PMID- 16380513 TI - MAP kinase phosphatase 1 controls innate immune responses and suppresses endotoxic shock. AB - Septic shock is a leading cause of morbidity and mortality. However, genetic factors predisposing to septic shock are not fully understood. Excessive production of proinflammatory cytokines, particularly tumor necrosis factor (TNF) alpha, and the resultant severe hypotension play a central role in the pathophysiological process. Mitogen-activated protein (MAP) kinase cascades are crucial in the biosynthesis of proinflammatory cytokines. MAP kinase phosphatase (MKP)-1 is an archetypal member of the dual specificity protein phosphatase family that dephosphorylates MAP kinase. Thus, we hypothesize that knockout of the Mkp-1 gene results in prolonged MAP kinase activation, augmented cytokine production, and increased susceptibility to endotoxic shock. Here, we show that knockout of Mkp-1 substantially sensitizes mice to endotoxic shock induced by lipopolysaccharide (LPS) challenge. We demonstrate that upon LPS challenge, Mkp-1 /- cells exhibit prolonged p38 and c-Jun NH2-terminal kinase activation as well as enhanced TNF-alpha and interleukin (IL)-6 production compared with wild-type cells. After LPS challenge, Mkp-1 knockout mice produce dramatically more TNF alpha, IL-6, and IL-10 than do wild-type mice. Consequently, Mkp-1 knockout mice develop severe hypotension and multiple organ failure, and exhibit a remarkable increase in mortality. Our studies demonstrate that MKP-1 is a pivotal feedback control regulator of the innate immune responses and plays a critical role in suppressing endotoxin shock. PMID- 16380515 TI - 2-methoxyestradiol induces cell cycle arrest and mitotic cell apoptosis in human vascular smooth muscle cells. AB - It has been shown that 2-methoxyestradiol (2-ME) inhibits cell proliferation and DNA synthesis in human aortic smooth muscle cells. However, the cellular mechanisms underlying the antiproliferative activity of 2-ME are unclear. The present study was performed to explore the cellular mechanisms whereby 2-ME leads to growth inhibition and apoptosis of human smooth muscle cells. Our results demonstrate that at 1 hour of treatment, 1 micromol/L 2-ME induces multiple spindles, overamplified centrosomes, and multipolar cytokinesis, whereas 10 micromol/L 2-ME causes completely damaged spindle, disorientated centrosomes, and missegregated chromosomes. At 6 hours of treatment, the mitotic index was increased and reached a maximal level, and cells with 4N DNA content (4N cells) began to accumulate. The increased mitotic cells induced by 2-ME were apoptotic as detected by both annexin V and TUNEL staining. Blockage of cells in G(1/0) phase by thymidine prevented 2-ME-induced apoptosis. In addition, the increased mitotic index declined concurrently when even more 4N cells accumulated at 12 to 48 hours of treatment with 10 micromol/L 2-ME. Furthermore, in response to 2-ME, cells delayed entry into the next cell cycle and exhibited aneuploidy or micronuclei. Some aneuploidy cells continued to synthesize DNA. We conclude that 2-ME treatment not only arrests cells in mitosis and promotes mitotic cell apoptosis, but also causes cells to undergo "mitotic slippage" and endoreduplication. The induction of mitotic cell arrest and apoptosis may be a major cellular mechanism by which 2-ME inhibits proliferation of human smooth muscle cells. PMID- 16380516 TI - Does obesity induce resistance to the long-term cardiovascular and metabolic actions of melanocortin 3/4 receptor activation? AB - Previous studies suggest that blockade of melanocortin 3 and 4 receptors (MC3/4 R) markedly attenuates the chronic hypertensive effects of leptin. Although obesity has been reported to be associated with leptin "resistance," it is unclear whether obesity alters the cardiovascular and metabolic effects of chronic MC3/4-R activation. Therefore, we tested whether the cardiovascular and metabolic actions of MC3/4-R activation are attenuated in Sprague-Dawley rats fed a high-fat diet (HF, n=6) compared with rats fed a standard chow (NF, n=6) for 12 months. A 21G steel cannula was placed in the lateral ventricle for ICV infusion, and arterial and venous catheters were implanted for measurement of mean arterial pressure (MAP) 24 hours/day and IV infusions. After a 5-day control period, rats were infused with MC3/4-R agonist melanotan II (10 ng/h, ICV), for 10 days followed by a 5-day recovery period. HF rats were heavier (558+/-21 versus 485+/ 13 g) with 140% more visceral fat than NF rats, hyperleptinemic (8.9+/-0.5 versus 2.7+/-0.5 ng/mL), and insulin resistant. HF rats also had higher MAP (109+/-3 versus 100+/-1 mm Hg). Chronic melanotan II infusion significantly increased MAP in HF and NF (7+/-2 and 6+/-1 mm Hg), decreased caloric intake (-32+/-2 and -25 +/-2 kcal/day), and reduced insulin levels in both groups by approximately 50%. Thus, the metabolic and cardiovascular actions of chronic MC3/4-R activation are preserved in diet-induced obesity, supporting a potential role for the hypothalamic melanocortin system in obesity hypertension. PMID- 16380517 TI - Adenovirus-mediated small-interference RNA for in vivo silencing of angiotensin AT1a receptors in mouse brain. AB - Because of the lack of pharmacological approaches, molecular genetic methods have been required to differentiate between angiotensin type 1(AT1) receptor subtypes AT1a and AT1b. RNA interference is a new tool for the study of gene function, producing specific downregulation of protein expression. In this study, we used the small hairpin RNA (shRNA) cassette method to screen target sites for selectively silencing AT1a or AT1b receptor subtypes in cultured Neuro-2a cells using real-time RT-PCR. For in vivo functional studies, we used C57BL mice with arterial telemetric probes and computerized licking monitors to test the effect of adenovirus carrying the DNA sequence coding AT1a shRNA (Ad-AT1a-shRNA). Ad AT1a-shRNA was injected into the lateral ventricle (intracerebroventricular) or the brain stem nucleus tractus solitaries/dorsal vagal nucleus (NTS/DVN) with measurement of water intake, blood pressure (BP), and heart rate (HR) for up to 20 days after injection. Tissue culture studies verified the specificity and the efficiency of the constructs. In animal studies, beta-galactosidase staining and Ang receptor binding assays showed expression of shRNA and downregulation of Ang AT1 receptors in the subfornical organ and NTS/DVN by >70%. Intracerebroventricular injection of Ad-AT1a-shRNA increased water intake with no effect on BP or HR. In contrast, microinjection of Ad-AT1a-shRNA into NTS/DVN caused a decrease in BP with no effect on HR or water intake. Results demonstrate the use of the RNA interference method in site-directed silencing of gene expression and provide a method for the in vivo study of Ang AT1 receptor function. PMID- 16380518 TI - Injection of nerve growth factor into stellate ganglia improves norepinephrine reuptake into failing hearts. AB - An impairment of cardiac norepinephrine reuptake through the neuronal norepinephrine transporter promotes depletion of cardiac norepinephrine stores and local cardiac sympathetic activation in heart failure. Nerve growth factor regulates differentiation and survival of adult sympathetic cells and is decreased in failing hearts. We hypothesized that injection of nerve growth factor into stellate ganglia normalizes cardiac norepinephrine homeostasis in experimental heart failure. Rats with transverse aortic constriction characterized by heart failure, depleted cardiac norepinephrine stores, and impaired cardiac norepinephrine reuptake were used as an experimental model. Nerve growth factor (20 microg) or saline was directly injected into left stellate ganglia 4 weeks after transverse aortic constriction. Thirty-two hours after injection, determinants of cardiac norepinephrine homeostasis were measured. As compared with saline, nerve growth factor refilled depleted cardiac norepinephrine stores and improved cardiac [3H]-norepinephrine uptake into isolated perfused hearts of transverse aortic constricted rats. In addition, pharmacological blockade of the norepinephrine transporter led to a higher increase in the overflow of endogenous norepinephrine from hearts of nerve growth factor-injected than saline-injected transverse aortic constricted rats. Norepinephrine transporter mRNA levels and the density of cardiac sympathetic nerves were not changed. Thirty-two hours after nerve growth factor injection, echocardiography revealed an increase in fractional shortening as compared with 2 days before injection. In conclusion, nerve growth factor attenuates local cardiac sympathetic overdrive of hypertrophic hearts by improving cardiac norepinephrine reuptake and might represent a novel therapeutic principle in the treatment of heart failure. PMID- 16380519 TI - Time course of maternal plasma volume and hormonal changes in women with preeclampsia or fetal growth restriction. AB - We tested the hypothesis that women with idiopathic fetal growth restriction (FGR) or preeclampsia (PE) have lower concentrations of some water-retaining hormones, such as aldosterone and estradiol, either preceding or concomitant with the onset of the reduced plasma volume described in these women. Plasma volume and serum concentrations of estradiol, progesterone, and aldosterone were measured serially at monthly intervals in 135 pregnant women from week 10 until term. Twenty-three developed idiopathic FGR, 17 had PE, and 95 remained normotensive and delivered normal-size infants (controls). Changes over time for each variable were studied using mixed models. Maternal age, parity, and weight/height at term were similar in all of the groups. Birth weight, body length, and ponderal index were lower in FGR and PE than in controls. Plasma volume increased throughout pregnancy in controls but was lower in FGR and PE from week 14 to 17 until term. Aldosterone values were lower in PE from week 26 to 29 onwards and in FGR after week 34. Progesterone concentrations were higher in PE than either control or FGR from week 18 to 21 until term. In contrast, FGR pregnancies had reduced progesterone and estradiol concentrations after week 34. Progesterone:estradiol ratio was significantly higher only in the PE group. In mothers with idiopathic FGR or PE, less expansion in plasma volume occurred before alterations in hormonal concentrations. We speculate that the early rise in progesterone may have a pathogenic role in the development of preeclampsia. PMID- 16380520 TI - Uric acid, left ventricular mass index, and risk of cardiovascular disease in essential hypertension. AB - Elevated serum uric acid (UA) is frequently encountered in individuals with hypertension, but whether the relationship between UA and cardiovascular events is circumstantial or causal remains to be answered. We examined the association between serum UA and left ventricular mass index (LVMI) and investigated prospectively whether the combination of UA and LVMI can predict the incidence of cardiovascular disease (CVD) in asymptomatic subjects with essential hypertension. A total of 619 subjects (mean age, 61 years; 52% female) free of prior CVD were included in this study. A significant association between UA and LVMI was also confirmed in multiple regression analysis (male: F=4.29, P<0.04; female: F=4.24, P<0.05). During follow-up (mean, 34 months), 28 subjects (14 female) developed CVD including myocardial infarction, angina pectoris, congestive heart failure, cerebral infarction, and transient cerebral ischemia. Sex-specific median values were used to separate the higher group from the lower group of UA and LVMI. Kaplan-Meier curves showed a significantly poorer survival rate in the group with higher UA and LVMI (LVMI, male: >126.9, female: >112.0 g/m2; UA, male: >374.7, female: >303.3 micromol/L; log-rank chi2=13.18; P<0.01). Multivariate Cox regression analysis showed that the combination of higher UA and LVMI was an independent predictor for CVD events (hazard ratio, 2.38; P<0.03). Our findings demonstrate that UA is independently associated with LVMI and suggest that the combination of hyperuricemia combined with left ventricular hypertrophy is an independent and powerful predictor for CVD. The association between UA and CVD events may be introduced in part because of a direct association of UA with LVMI. PMID- 16380521 TI - Problems with blood pressure terminology. PMID- 16380522 TI - Complex relationship between blood pressure and mortality in type 2 diabetic patients: a follow-up of the Botnia Study. AB - The presence of hypertension aggravates the high cardiovascular risk in type 2 diabetic patients. Pulse pressure is a marker of arterial stiffness and constitutes a risk factor for cardiovascular mortality. This study examines the relationship between different blood pressure indices and mortality in a cohort of type 2 diabetic patients. A total of 1294 type 2 diabetic patients with a median age of 69.1 years participated in the Botnia Study from 1990 to 1997. In 2004, after a median follow-up of 9.5 years, data on mortality was collected from the national population registry and hospital records. Systolic and diastolic blood pressure correlated negatively with mortality after adjustment for other risk factors. The association between low systolic and diastolic blood pressure and mortality was pronounced in patients with previous cardiovascular disease. A U-shaped association between pulse pressure and mortality was observed in elderly patients. These observations could be linked to arterial stiffness and heart failure. Low blood pressure in high-risk patients is likely to be a marker of poor health rather than the cause of mortality. The results suggest that the role of blood pressure as a risk marker in elderly type 2 diabetic patients with cardiovascular disease needs to be reevaluated. PMID- 16380523 TI - Aortic pulse wave velocity is associated with the presence and quantity of coronary artery calcium: a community-based study. AB - We investigated the relationship of aortic pulse wave velocity (aPWV), a measure of central arterial stiffness, with the presence and quantity of coronary artery calcium (CAC) in a community-based sample of adults without prior history of heart attack or stroke (n=401, mean age 59.8 years, 53% men). ECG-gated waveforms of the right carotid and right femoral artery were obtained by applanation tonometry, and aPWV was calculated using established methods. CAC was measured noninvasively by electron beam computed tomography, and CAC score was calculated. aPWV was significantly correlated with log(CAC +1; r=0.41; P<0.0001) and pulse pressure (r=0.47; P<0.0001). Multivariable logistic and linear regression models were used to identify independent predictors of the presence and quantity of CAC, respectively. In multivariable logistic regression analyses, aPWV was associated with the presence of CAC (P=0.011) after adjustment for age, male sex, total cholesterol, high-density lipoprotein cholesterol, diabetes, history of smoking, systolic blood pressure, body mass index, and use of hypertension and statin medications. In multivariable linear regression analyses, aPWV was significantly associated with log(CAC +1) after adjustment for the covariates enumerated above (P<0.0001). aPWV remained significantly associated with both the presence and quantity of CAC even after the additional adjustment for diastolic blood pressure. We conclude that aPWV is related to subclinical coronary atherosclerosis independent of conventional risk factors (including indices of blood pressure) and may be a biomarker of cardiovascular risk in asymptomatic individuals. PMID- 16380524 TI - Weather-related changes in 24-hour blood pressure profile: effects of age and implications for hypertension management. AB - A downward titration of antihypertensive drug regimens in summertime is often performed on the basis of seasonal variations of clinic blood pressure (BP). However, little is known about the actual interaction between outdoor air temperature and the effects of antihypertensive treatment on ambulatory BP. The combined effects of aging, treatment, and daily mean temperature on clinic and ambulatory BP were investigated in 6404 subjects referred to our units between October 1999 and December 2003. Office and mean 24-hour systolic BP, as well as morning pressure surge, were significantly lower in hot (>90th percentiles of air temperature; 136+/-19, 130+/-14, and 33.3+/-16.1 mm Hg; P<0.05 for all), and higher in cold (<10th percentiles) days (141+/-12, 133+/-11, and 37.3+/-9.5 mm Hg; at least P<0.05 for all) when compared with intermediate days (138+/-18, 132+/-14, and 35.3+/-15.4 mm Hg). At regression analysis, 24-hour and daytime systolic pressure were inversely related to temperature (P<0.01 for all). Conversely, nighttime systolic pressure was positively related to temperature (P<0.02), with hot days being associated with higher nighttime pressure. Air temperature was identified as an independent predictor of nighttime systolic pressure increase in the group of elderly treated hypertensive subjects only. No significant relationship was found between air temperature and heart rate. Our results show for the first time that hot weather is associated with an increase in systolic pressure at night in treated elderly hypertensive subjects. This may be because of a nocturnal BP escape from the effects of a lighter summertime drug regimen and may have important implications for seasonal modulation of antihypertensive treatment. PMID- 16380525 TI - Caution for winter morning surge in blood pressure: a possible link with cardiovascular risk in the elderly. PMID- 16380526 TI - Retinal vessel diameters and risk of hypertension: the Rotterdam Study. AB - Generalized retinal arteriolar narrowing is an important sign of systemic hypertension, and a lower arteriolar:venular diameter ratio predicts the risk of hypertension. We investigated whether this association was based on arteriolar or venular diameters or both. This study was based on the prospective population based Rotterdam Study (1990-1993) and included 1900 participants (> or =55 years of age) of whom 739 persons had normal blood pressure (systolic <120 mm Hg and diastolic <80 mm Hg) and 1161 prehypertension (systolic 120 to 139 mm Hg or diastolic 80 to 89 mm Hg). For each participant, retinal arteriolar and venular diameters were measured on digitized images of 1 eye. After a mean follow-up of 6.6 years, 808 persons developed hypertension, defined as either systolic blood pressure > or =140 mm Hg or diastolic blood pressure > or =90 mm Hg or use of antihypertensive medication. Adjusted for age, gender, follow-up time, body mass index, smoking, diabetes mellitus, total and high-density lipoprotein cholesterol, C-reactive protein, and intima-media thickness, arteriolar narrowing was associated with an increased risk of hypertension (odds ratio per SD: 1.38; 95% CI, 1.23 to 1.55); for venular narrowing this was less striking (OR: 1.17; 95% CI, 1.04 to 1.32). Each SD decrease in the arteriolar:venular diameter ratio significantly increased the risk of hypertension by 24%. To examine the effect of baseline blood pressure, we stratified persons into those with "normal blood pressure" or "prehypertension." Within these strata, arteriolar narrowing was still related to incident hypertension. These data show that both retinal arteriolar and venular narrowing may precede the development of systemic hypertension. PMID- 16380527 TI - Risk factors for arterial hypertension in adults with initial optimal blood pressure: the Strong Heart Study. AB - Whether metabolic factors and their change over time influence development of arterial hypertension in adults with initially optimal blood pressure (BP) is unknown. We analyzed associations of BP in the optimal range (<120/80 mm Hg), metabolic risk factors, and their changes over 4-year follow-up, with 8-year incident hypertension, in a cohort of American Indians with a high prevalence of obesity. At baseline, 967 participants with optimal BP and no prevalent cardiovascular disease (69.5% women; mean age, 54+/-7 years) were evaluated and reexamined after 4 (second examination) and 8 years to evaluate predictors of 8 year incident arterial hypertension. In participants with normal glucose tolerance, baseline BP and decrease in high-density lipoprotein cholesterol from baseline to the second examination were the most potent predictors of 8-year arterial hypertension (both P<0.0001), with additional effects of baseline waist circumference and its increase, increase in BP, and presence of diabetes at the second examination (all P<0.04). In participants with impaired glucose tolerance or diabetes, the most potent predictor of 8-year incident hypertension was diabetes at the second examination (P<0.0001) followed by a increase in BP and LDL cholesterol over the first 4 years (both P<0.001). Thus, incident arterial hypertension can be predicted by initial metabolic profile and unfavorable metabolic variations over time, in addition to initial BP. At optimal levels of initial BP, increasing abdominal obesity, and abnormal lipid profile are major predictors of development of arterial hypertension. Possible implications of these findings for primary cardiovascular prevention should be tested in prospective studies. PMID- 16380528 TI - Weight and mortality. PMID- 16380530 TI - Leptin regulates cardiomyocyte contractile function through endothelin-1 receptor NADPH oxidase pathway. AB - Leptin, the obese gene product, plays an important role in the regulation of cardiac function. However, the mechanism behind leptin-induced cardiomyocyte contractile response is poorly understood. This study was designed to examine whether endothelin-1 receptor and NADPH oxidase play any role in leptin-induced cardiac contractile response. Isolated murine cardiomyocytes were exposed to leptin (5, 50, and 100 nmol/L) for 60 minutes in the absence or presence of the ETA receptor antagonist BQ123 (1 micromol/L), the ETB receptor antagonist BQ788 (1 micromol/L), or the NADPH oxidase inhibitor apocynin (100 micromol/L) before mechanical function was studied. Superoxide levels were measured by dihydroethidium fluorescent dye and the superoxide dismutase-inhibitable reduction of cytochrome c. NADPH oxidase subunit expression (p22phox, p47phox, p67phox, and gp91phox) was evaluated with Western blot. Leptin depressed peak shortening and maximal velocity of shortening/relengthening (+/-dL/dt), prolonged the duration of relengthening (TR90) without affecting the time-to-peak cell shortening. Consistent with the mechanical characteristics, myocytes treated with leptin displayed a reduced electrically stimulated rise in intracellular Ca2+ (change in fura-2 fluorescence intensity) associated with a prolonged intracellular Ca2+ decay rate. All of the abnormalities were significantly attenuated by apocynin, BQ123, or BQ788. Intracellular superoxide generation was enhanced after leptin treatment, which was partially blocked by apocynin, BQ123, or BQ788. Leptin had no effect on p22phox and gp91phox but upregulated protein expression of p67phox and p47phox, both of which were inhibited by apocynin, BQ123, or BQ788. These results suggest that leptin suppresses cardiac contractile function in ventricular myocytes through the endothelin-1 receptor and NADPH oxidase-mediated pathway. PMID- 16380531 TI - Knockdown of arginase I restores NO signaling in the vasculature of old rats. AB - Arginase, expressed in endothelial cells and upregulated in aging blood vessels, competes with NO synthase (NOS) for l-arginine, thus modulating vasoreactivity and attenuating NO signaling. Moreover, arginase inhibition restores endothelial NOS signaling and l-arginine responsiveness in old rat aorta. The arginase isoform responsible for modulating NOS, however, remains unknown. Because isoform specific arginase inhibitors are unavailable, we used an antisense (AS) oligonucleotide approach to knockdown arginase I (Arg I). Western blot and quantitative PCR confirmed that Arg I is the predominant isoform expressed in endothelialized aortic rings and is upregulated in old rats compared with young. Aortic rings from 22-month-old rats were incubated for 24 hours with sense (S), AS oligonucleotides, or medium alone (C). Immunohistochemistry, immunoblotting, and enzyme assay confirmed a significant knockdown of Arg I protein and arginase activity in AS but not S or C rings. Conversely, calcium-dependent NOS activity and vascular metabolites of NO was increased in AS versus S or C rings. Acetylcholine (endothelial-dependent) vasorelaxant responses were enhanced in AS versus S or C treated rings. In addition, 1H-oxadiazolo quinoxalin-1-one (10 micromol/L), a soluble guanylyl cyclase inhibitor, increased the phenylephrine response in AS compared with S and C rings suggesting increased NO bioavailability. Finally, l-arginine (0.1 mmol/L)-induced relaxation was increased in AS versus C rings. These data support our hypothesis that Arg I plays a critical role in the pathobiology of age-related endothelial dysfunction. AS oligonucleotides may, therefore, represent a novel therapeutic strategy against age-related vascular endothelial dysfunction. PMID- 16380532 TI - Prognostic relevance of blood pressure variability. PMID- 16380533 TI - Prognostic significance for stroke of a morning pressor surge and a nocturnal blood pressure decline: the Ohasama study. AB - There is continuing controversy over whether the pattern of circadian blood pressure (BP) variation that includes a nocturnal decline in BP and a morning pressor surge has prognostic significance for stroke risk. In this study, we followed the incidence of stroke in 1430 subjects aged > or =40 years in Ohasama, Japan, for an average of 10.4 years. The association between stroke risk and the pattern of circadian BP variation was analyzed with a Cox proportional hazards model after adjustment for possible confounding factors. There was no significant association between total stroke risk and the nocturnal decline in BP (percentage decline from diurnal level) or between total stroke risk and the morning pressor surge. The cerebral infarction risk was significantly higher in subjects with a <10% nocturnal decline in BP as compared with subjects who had a > or =10% nocturnal decline in BP (P=0.04). The morning pressor surge was not associated with a risk of cerebral infarction. On the other hand, an increased risk of cerebral hemorrhage was observed in subjects with a large morning pressor surge (> or =25 mm Hg; P=0.04). Intracerebral hemorrhage was also observed more frequently in extreme dippers (those with a > or =20% nocturnal decline in BP) than dippers (those with a 10% to 19% decline; P=0.02). A disturbed nocturnal decline in BP is associated with cerebral infarction, whereas a large morning pressor surge and a large nocturnal decline in BP, which are analogous to a large diurnal increase in BP, are both associated with cerebral hemorrhage. PMID- 16380534 TI - Integrins in hypertensive remodeling. PMID- 16380535 TI - Synergistic acceleration of arterial stiffening in the presence of raised blood pressure and raised plasma glucose. AB - Arterial stiffness is recognized as a marker of arterial damage and an indicator of cardiovascular risk. This observational study was conducted to examine the synergistic effect of raised blood pressure (RBP; > or =130/85 mm Hg) and raised plasma glucose (RPG; > or =110 mg/dL) even at levels below those conventionally used to define hypertension and diabetes on the rate of increase of the pulse wave velocity (PWV) over a 3-year period in 2080 Japanese men (age 42+/-9 years). First, the subjects were classified into 4 groups based on the presence at the first examination of RBP, RPG, both abnormalities, or neither abnormality. The estimated annual rate of increase of the PWV was higher in subjects with both the abnormalities than in those with either abnormality alone or neither of the 2 abnormalities. Second, the subjects were also classified based on the evolutional status of these abnormalities during the study period; persistence of both of the abnormalities synergistically accelerated the rate of increase of the PWV (68.3+/ 7.1 cm/s per year), as compared with the persistence of either abnormality alone (persistence of RBP alone: 18.2+/-1.6 cm/s per year; persistence of RPG alone: 21.2+/-7.4 cm/s per year) or persistence of neither abnormality (11.1+/-0.8 cm/s per year; P<0.01). Thus, blood pressure and fasting plasma glucose levels even below those defining hypertension and diabetes may synergistically lead to progression of arteriosclerotic arterial damage. This synergistic progression may contribute to the additive increases in the risk of cardiovascular events, at least in part. PMID- 16380536 TI - alphaV integrins are necessary for eutrophic inward remodeling of small arteries in hypertension. AB - Human essential hypertension is characterized by eutrophic remodeling of small arteries, with little evidence of hypertrophy. Likewise, vessels of young hypertensive TGR(mRen2)27 animals have undergone similar structural alterations. The role of integrins in resistance arteries of TGR(mRen2)27 during the eutrophic remodeling process was examined as blood pressure rose. Initially, 8 alpha and 3 beta integrins were identified and levels of expression investigated using RT PCR. As pressure increased and remodeling advanced, integrin expression profiles revealed that only alphaV was significantly raised. In conjunction, we confirmed elevated integrin alphaV protein levels in TGR(mRen2)27 rat arteries and localization to the media using immunofluorescence. beta1 and beta3, but not beta5 integrin subunits were coprecipitated with integrin alphaV and are implicated in the eutrophic remodeling process. Administration of a peptide antagonist of alphaVbeta3 abolished remodeling but enhanced growth, indicating that hypertrophy supervened as a response to hypertension-induced increases in wall stress. We have established that the only upregulated integrin, the alphaV subunit of integrin alphaVbeta3, has a crucial role in the hypertensive remodeling process of TGR(mRen2)27 rat resistance arteries. During hypertensive remodeling, functions of specific alphaVbeta3-extracellular matrix interactions are likely to allow vascular smooth muscle cell-length autoregulation, which includes a migratory process, to maintain a narrowed lumen after a prolonged constricted state. PMID- 16380537 TI - Angiotensin-II type 1 receptor-mediated hypertension in D4 dopamine receptor deficient mice. AB - Dopamine receptors are important in systemic blood pressure regulation. D4 receptors are expressed in the kidney and brain, but their role in cardiovascular regulation is unknown. In pentobarbital-anesthetized mice, systolic and diastolic blood pressures were elevated in sixth-generation D4 receptor-deficient (D4(-/-)) mice and in tenth-generation D4(-/-) mice compared with D4 wild-type (D4(+/+)) littermates. The conscious blood pressures measured via a chronic arterial (femoral) catheter or telemetry (carotid) were also higher in D4(-/-) mice than in D4 littermates. Basal renal and plasma renin concentrations were similar in the 2 mouse strains. The protein expression of angiotensin II type 1 receptor was increased in homogenates of kidney (330+/-53%, n=5) and brain (272+/-69%, n=5) of D4(-/-) mice relative to D4(+/+) mice (kidney: 100+/-12%, n=5; brain: 100+/-32%, n=5). The expression of the receptor in renal membrane was also increased in D4( /-) mice (289+/-28%, n=8) relative to D4(+/+) mice (100+/-14%, n=10). In contrast, the expression in the heart was similar in the 2 strains. Bolus intravenous injection of angiotensin II type 1 receptor antagonist losartan initially decreased mean arterial pressures to a similar degree in D4(-/-) and D4(+/+) littermates. However, the hypotensive effect of losartan dissipated after 10 minutes in D4(+/+) mice, whereas the effect persisted for >45 minutes in D4(-/ ) mice. We conclude that the absence of the D(4) receptor increases blood pressure, possibly via increased angiotensin II type 1 receptor expression. PMID- 16380538 TI - Activation of AMP-activated protein kinase enhances angiotensin ii-induced proliferation in cardiac fibroblasts. AB - AMP-activated kinase (AMPK) is a highly conserved heterotrimeric kinase that functions as a metabolic regulator of cellular enzymes involved in carbohydrate and fat metabolism, which regulate ATP conservation and synthesis. Here, we investigated whether AMPK signaling has a role in the regulation of angiotensin II (Ang II)-induced proliferation in rat cardiac fibroblasts. Aminoimidazole-4 carboxamide-1-beta-ribofuranoside (AICAR) activated AMPK in rat cardiac fibroblasts and increased Ang II-induced extracellular signal-regulated kinase 1/2 phosphorylation and activity. AICAR also increased Ang II-induced c-fos mRNA expression in the cells. [3H]-thymidine and [3H]-proline incorporation by cardiac fibroblasts treated with Ang II was enhanced when the cells were pretreated with AICAR. Inhibition of AMPK by small interfering RNA for AMPKalpha1 suppressed Ang II-induced extracellular signal-regulated kinase activity, c-fos mRNA expression, and cell proliferation. Treatment of rats with AICAR (1 mg/g body weight per day) for 1 week significantly enhanced Ang II-induced hypertrophy of the myocardium. Our findings indicate that AMPK works as a stimulator of the Ang II-induced proliferative pathway in cardiac fibroblasts. Inhibition of AMPK signaling might serve as a new therapeutic target of remodeling of the hypertrophic myocardium. PMID- 16380539 TI - Extracellular ATP stimulates NO production in rat thick ascending limb. AB - NO produced by NO synthase (NOS) 3 acts as an autacoid to regulate NaCl absorption in the thick ascending limb. ATP induces NO production by NOS 3 in endothelial cells. We hypothesized that extracellular ATP activates NOS in thick ascending limbs through P2 receptors. To test this, we measured intracellular NO production using the NO-selective fluorescent dye DAF-2 in suspensions of rat medullary thick ascending limbs. We found that ATP increased DAF-2 fluorescence in a concentration-dependent manner, reaching saturation at &200 micromol/L with an EC50 of 37 micromol/L. The increase was blunted by 74% by the nonselective NOS inhibitor L-omega-nitro-arginine-methyl-ester (2 mmol/L; 60+/-7 versus 16+/-6 arbitrary fluorescence units; P<0.02; n=5). In the presence of the P2 receptor antagonist suramin (300 micromol/L), ATP-induced NO production was reduced by 64% (101+/-11 versus 37+/-5 arbitrary fluorescence units; P<0.002; n=5). Blocking ATP hydrolysis with a 5'-ectonucleotidase inhibitor, ARL67156 (30 micromol/L) enhanced the response to ATP and shifted the EC(50) to 0.8 micromol/L. In the presence of ARL67156, the EC50 of the P2X-selective agonist beta,gamma-methylene adenosine 5'-triphosphate was 4.8 micromol/L and the EC50 for the P2Y-selective agonist UTP was 40.4 micromol/L. The maximal responses for both agonists were similar. Taken together, these data indicate that ATP stimulates NO production in the thick ascending limb primarily through P2X receptor activation and that ATP hydrolysis may regulate NO production. PMID- 16380540 TI - Renal angiotensin type 2 receptors mediate natriuresis via angiotensin III in the angiotensin II type 1 receptor-blocked rat. AB - Whereas angiotensin (Ang) II is the major effector peptide of the renin angiotensin system, its metabolite, des-aspartyl1-Ang II (Ang III), may also have biologic activity. We investigated the effects of renal interstitial (RI) administration of candesartan (CAND), a specific Ang II type 1 receptor (AT1) blocker, with and without coinfusion of PD-123319 (PD), a specific Ang II type 2 receptor (AT2) blocker, on Na+ excretion (UNaV) in uninephrectomized rats. We also studied the effects of unilateral RI infusion of Ang II or Ang III on UNaV with and without systemic infusion of CAND with the noninfused kidney as control. In rats receiving normal Na+ intake, RI CAND increased UNaV from 0.07+/-0.08 to 0.82+/-0.17 micromol/min (P<0.01); this response was abolished by PD. During Na+ restriction, CAND increased UNaV from 0.06+/-0.02 to 0.1+/-0.02 micromol/min (P<0.05); this response also was blocked by PD. In rats with both kidneys intact, in the absence of CAND, unilateral RI infusion of Ang III did not significantly alter UNaV. However, with systemic CAND infusion, RI Ang III increased U(Na)V from 0.08+/-0.01 micromol/min to 0.18+/-0.04 micromol/min (P<0.01) at 3.5 nmol/kg per minute, and UNaV remained elevated throughout the infusion; this response was abolished by PD. However, RI infusion of Ang II did not significantly alter UNaV at any infusion rate (3.5 to 80 nmol/kg per minute) with or without systemic CAND infusion. These results suggest that intrarenal AT1 receptor blockade engenders natriuresis by activation of AT2 receptors. AT2 receptor activation via Ang III, but not via Ang II, mediates the natriuretic response in the presence of systemic AT1 receptor blockade. PMID- 16380541 TI - Pregnancy prevents hypertensive remodeling of cerebral arteries: a potential role in the development of eclampsia. AB - We investigated how hypertension during pregnancy affected passive structural (wall:lumen, wall stress) and active (myogenic activity) responses of the cerebral circulation. Female nonpregnant (NP; n=8) Sprague Dawley rats were compared with late-pregnant (LP; day 19 to 20, n=6) rats. Some animals were treated with the NO synthase inhibitor nitro-L-arginine in their drinking water to raise blood pressure. LP rats (n=6) were treated for the last 7 days of pregnancy (last trimester) to mimic preeclampsia and compared with NP rats treated for the same duration (n=8). Active and passive responses were determined on isolated and pressurized third-order posterior cerebral arteries. Nitro-L arginine treatment significantly raised blood pressure in both groups of animals that was associated with increased wall thickness and wall:lumen ratio in the NP hypertensive animals versus controls (P<0.05). In contrast, this response to pressure was absent in LP animals, which had similar wall measurements. In addition, arteries from NP hypertensive animals had increased myogenic tone and pressure of forced dilatation compared with NP control animals (P<0.01). Again, this response was lacking in the LP hypertensive animals that had similar tone and pressure of forced dilatation as normotensive controls. The increased tone and wall thickness decreased wall stress in the NP hypertensive animals, a response that did not occur in LP hypertensive animals. Because medial hypertrophy is considered a protective response to elevated blood pressure, these results suggest that hypertension in pregnancy may predispose the cerebral circulation to autoregulatory breakthrough and blood-brain-barrier disruption when blood pressure is elevated, as during eclampsia. PMID- 16380542 TI - Obesity and cardiovascular disease: pathophysiology, evaluation, and effect of weight loss: an update of the 1997 American Heart Association Scientific Statement on Obesity and Heart Disease from the Obesity Committee of the Council on Nutrition, Physical Activity, and Metabolism. AB - Obesity is becoming a global epidemic in both children and adults. It is associated with numerous comorbidities such as cardiovascular diseases (CVD), type 2 diabetes, hypertension, certain cancers, and sleep apnea/sleep-disordered breathing. In fact, obesity is an independent risk factor for CVD, and CVD risks have also been documented in obese children. Obesity is associated with an increased risk of morbidity and mortality as well as reduced life expectancy. Health service use and medical costs associated with obesity and related diseases have risen dramatically and are expected to continue to rise. Besides an altered metabolic profile, a variety of adaptations/alterations in cardiac structure and function occur in the individual as adipose tissue accumulates in excess amounts, even in the absence of comorbidities. Hence, obesity may affect the heart through its influence on known risk factors such as dyslipidemia, hypertension, glucose intolerance, inflammatory markers, obstructive sleep apnea/hypoventilation, and the prothrombotic state, in addition to as-yet-unrecognized mechanisms. On the whole, overweight and obesity predispose to or are associated with numerous cardiac complications such as coronary heart disease, heart failure, and sudden death because of their impact on the cardiovascular system. The pathophysiology of these entities that are linked to obesity will be discussed. However, the cardiovascular clinical evaluation of obese patients may be limited because of the morphology of the individual. In this statement, we review the available evidence of the impact of obesity on CVD with emphasis on the evaluation of cardiac structure and function in obese patients and the effect of weight loss on the cardiovascular system. PMID- 16380543 TI - Cyclooxygenase-1 deficiency in bone marrow cells increases early atherosclerosis in apolipoprotein E- and low-density lipoprotein receptor-null mice. AB - BACKGROUND: Cyclooxygenase-1 (COX-1) has been implicated in the pathogenesis of atherothrombosis and is expressed by the major cell types of atherosclerotic lesions. COX-1-mediated platelet thromboxane (TX) production has been proposed to promote both early atherosclerosis and thrombosis. Here, we examined the impact of COX-1 deficiency in bone marrow-derived cells on early atherogenesis in the mouse. METHODS AND RESULTS: LDL receptor (LDLR)(-/-) and apolipoprotein E (apoE)( /-) recipient mice were lethally irradiated and transplanted with COX-1(-/-) bone marrow. Mice reconstituted with COX-1(-/-) marrow had nearly complete (99.7%) loss of platelet TXA2 and significantly suppressed levels of macrophage and urinary TXA2 metabolites. Serum lipid levels and lipoprotein distributions did not differ between recipients reconstituted with COX-1(+/+) and COX-1(-/-) marrow. Surprisingly, the extent of atherosclerotic lesions in both LDLR(-/-) and apoE(-/-) mice reconstituted with COX-1(-/-) marrow was increased significantly compared with control mice transplanted with COX-1(+/+) marrow. Peritoneal macrophages isolated from LDLR(-/-) mice reconstituted with COX-1(-/-) marrow had increased lipopolysaccharide-induced levels of COX-2 mRNA and protein expression. Fetal liver cell transplantation studies revealed a 30% increase in atherosclerosis in COX-1(-/-)-->LDLR(-/-)mice compared with COX-1(+/+)-->LDLR(-/ )mice, whereas the extent of atherosclerosis was unchanged in COX-1(-/-)/COX-2(-/ )-->LDLR(-/-)mice. CONCLUSIONS: COX-1 deficiency in bone marrow-derived cells worsens early atherosclerosis in apoE(-/-) and LDLR(-/-) mice despite virtual elimination of platelet TX production. These data demonstrate that platelet TX production does not aggravate early atherosclerotic lesion formation and that upregulation of COX-2 expression in COX-1(-/-) macrophages is proatherogenic. PMID- 16380544 TI - Riboflavin lowers homocysteine in individuals homozygous for the MTHFR 677C->T polymorphism. AB - BACKGROUND: Meta-analyses predict that a 25% lowering of plasma homocysteine would reduce the risk of coronary heart disease by 11% to 16% and stroke by 19% to 24%. Individuals homozygous for the methylenetetrahydrofolate reductase (MTHFR) 677C-->T polymorphism have reduced MTHFR enzyme activity resulting from the inappropriate loss of the riboflavin cofactor, but it is unknown whether their typically high homocysteine levels are responsive to improved riboflavin status. METHODS AND RESULTS: From a register of 680 healthy adults 18 to 65 years of age of known MTHFR 677C-->T genotype, we identified 35 with the homozygous (TT) genotype and age-matched individuals with heterozygous (CT, n=26) or wild type (CC, n=28) genotypes to participate in an intervention in which participants were randomized by genotype group to receive 1.6 mg/d riboflavin or placebo for a 12-week period. Supplementation increased riboflavin status to the same extent in all genotype groups (8% to 12% response in erythrocyte glutathione reductase activation coefficient; P<0.01 in each case). However, homocysteine responded only in the TT group, with levels decreasing by as much as 22% overall (from 16.1+/-1.5 to 12.5+/-0.8 micromol/L; P=0.003; n=32) and markedly so (by 40%) in those with lower riboflavin status at baseline (from 22.0+/-2.9 and 13.2+/-1.0 micromol/L; P=0.010; n=16). No homocysteine response was observed in the CC or CT groups despite being preselected for suboptimal riboflavin status. CONCLUSIONS: Although previously overlooked, homocysteine is highly responsive to riboflavin, specifically in individuals with the MTHFR 677 TT genotype. Our findings might explain why this common polymorphism carries an increased risk of coronary heart disease in Europe but not in North America, where riboflavin fortification has existed for >50 years. PMID- 16380545 TI - CD8+ T lymphocytes regulate the arteriogenic response to ischemia by infiltrating the site of collateral vessel development and recruiting CD4+ mononuclear cells through the expression of interleukin-16. AB - BACKGROUND: Previous studies have demonstrated that macrophages and CD4+ T lymphocytes play pivotal roles in collateral development. Indirect evidence suggests that CD8+ T cells also play a role. Thus, after acute cerebral ischemia, CD8+ T cells infiltrate the perivascular space and secrete interleukin-16 (IL 16), a potent chemoattractant for monocytes and CD4+ T cells. We tested whether CD8+ T lymphocytes contribute to collateral vessel development and whether the lack of circulating CD8+ T cells prevents IL-16 expression, impairs CD4+ mononuclear cell recruitment, and reduces collateral vessel growth after femoral artery ligation in CD8(-/-) mice. METHODS AND RESULTS: After surgical excision of the femoral artery, laser Doppler perfusion imaging demonstrated reduced blood flow recovery in CD8(-/-) mice compared with C57/BL6 mice (ischemic/nonischemic limb at day 28, 0.66+/-0.04 versus 0.87+/-0.04, respectively; P<0.01). This resulted in greater calf muscle atrophy (mean fiber area, 785+/-68 versus 1067+/ 69 microm2, respectively; P<0.01) and increased fibrotic tissue content (10.8+/ 1.2% versus 7+/-1%, respectively; P<0.01). Moreover, CD8(-/-) mice displayed reduced IL-16 expression and decreased CD4+ T-cell recruitment at the site of collateral vessel development. Exogenous CD8+ T cells, infused into CD8(-/-) mice immediately after femoral artery ligation, selectively homed to the ischemic hind limb and expressed IL-16. The restoration of IL-16 expression resulted in significant CD4+ mononuclear cell infiltration of the ischemic limb, faster blood flow recovery, and reduced hindlimb muscle atrophy/fibrosis. When exogenous CD8+ T cells deficient in IL-16 (IL-16(-/-)) were infused into CD8(-/-) mice immediately after femoral artery ligation, they selectively homed to the ischemic hind limb but were unable to recruit CD4+ mononuclear cells and did not improve blood flow recovery. CONCLUSIONS: These results demonstrate that CD8+ T cells importantly contribute to the early phase of collateral development. After femoral artery ligation, CD8+ T cells infiltrate the site of collateral vessel growth and recruit CD4+ mononuclear cells through the expression of IL-16. Our study provides further evidence of the significant role of the immune system in modulating collateral development in response to peripheral ischemia. PMID- 16380546 TI - Relationship between C-reactive protein and subclinical atherosclerosis: the Dallas Heart Study. AB - BACKGROUND: Elevated levels of C-reactive protein (CRP) are associated with increased risk for incident cardiovascular events on the basis of observations from several prospective epidemiological studies. However, less is known regarding the relationship between CRP levels and atherosclerotic burden. METHODS AND RESULTS: We measured CRP in 3373 subjects 30 to 65 years of age who were participating in the Dallas Heart Study, a multiethnic, population-based, probability sample. Electron-beam CT scans were used to measure coronary artery calcification (CAC) in 2726 of these subjects, and MRI was used to measure aortic plaque in 2393. CRP levels were associated with most traditional cardiovascular risk factors. Subjects with CAC had higher median CRP levels than those without CAC (men: median, 2.4 versus 1.8 mg/L, P<0.001; women: median, 5.2 versus 3.6 mg/L, P<0.001), and there was a modest trend toward increasing CRP levels with increased CAC levels in men (P for trend=0.003) but not in women (P for trend=0.08). Male subjects with aortic plaque also had higher CRP levels than those without (median, 2.3 versus 1.8; P<0.001). In multivariate analysis adjusted for traditional cardiovascular risk factors, body mass index, and estrogen and statin medication use, the associations between CRP levels and CAC and CRP levels and aortic plaque were no longer statistically significant. CONCLUSIONS: In a large, population-based sample, subjects with higher CRP levels had a modest increase in the prevalence of subclinical atherosclerosis, but this association was not independent of traditional cardiovascular risk factors. CRP is a poor predictor of atherosclerotic burden. PMID- 16380547 TI - Increased small low-density lipoprotein particle number: a prominent feature of the metabolic syndrome in the Framingham Heart Study. AB - BACKGROUND: Levels of LDL cholesterol (LDL-C) are frequently not elevated in individuals with the metabolic syndrome (MetSyn). However, the atherogenic potential of LDL may depend on the number and size of LDL particles in addition to the cholesterol content of LDL. METHODS AND RESULTS: We examined the sex specific cross-sectional relations of small LDL particle number (determined by nuclear magnetic resonance spectroscopy) to the presence of MetSyn and its components in 2993 Framingham Heart Study participants (mean age, 51 years; 53% women) without cardiovascular disease (CVD) and the relations of small LDL particle number to CVD incidence in people with MetSyn. The MetSyn (> or =3 of 5 traits as defined by the National Cholesterol Education Adult Treatment Panel III) was present in 27% of men and 17% of women. In both sexes, small LDL particle number increased from 0 to 5 MetSyn traits, a pattern partly accounted for by strong correlations between small LDL particle number and serum triglycerides (r=0.61, P<0.0001) and HDL-C (r=-0.55, P<0.0001). Compared with participants without the MetSyn, those with the MetSyn had a higher CVD event rate. However, among participants with the MetSyn, CVD rates were similar for groups with an elevated versus a lower number of small LDL particles (defined by the sex-specific median). CONCLUSIONS: Small LDL particle number is elevated in the MetSyn, increases with the number of MetSyn components, and most prominently is correlated with triglycerides and HDL-C. Whereas increased small LDL particle number identified the MetSyn with high sensitivity, a higher small LDL particle number was not associated with greater CVD event rates in people with the MetSyn. PMID- 16380548 TI - Cardiac-specific overexpression of diacylglycerol kinase zeta prevents Gq protein coupled receptor agonist-induced cardiac hypertrophy in transgenic mice. AB - BACKGROUND: Diacylglycerol is a lipid second messenger that accumulates in cardiomyocytes when stimulated by Gqalpha protein-coupled receptor (GPCR) agonists such as angiotensin II, phenylephrine, and others. Diacylglycerol functions as a potent activator of protein kinase C (PKC) and is catalyzed by diacylglycerol kinase (DGK) to form phosphatidic acid and inactivated. However, the functional roles of DGK have not been previously examined in the heart. We hypothesized that DGK might prevent GPCR agonist-induced activation of diacylglycerol downstream signaling cascades and subsequent cardiac hypertrophy. METHODS AND RESULTS: To test this hypothesis, we generated transgenic (DGKzeta TG) mice with cardiac-specific overexpression of DGKzeta. There were no differences in heart size and heart weight between DGKzeta-TG and wild-type littermate mice. The left ventricular function was normal in DGKzeta-TG mice. Continuous administration of subpressor doses of angiotensin II and phenylephrine caused PKC translocation, gene induction of atrial natriuretic factor, and subsequent cardiac hypertrophy in WT mice. However, in DGKzeta-TG mice, neither translocation of PKC nor upregulation of atrial natriuretic factor gene expression was observed after angiotensin II and phenylephrine infusion. Furthermore, in DGKzeta-TG mice, angiotensin II and phenylephrine failed to increase cross-sectional cardiomyocyte areas and heart to body weight ratios. Phenylephrine-induced increases in myocardial diacylglycerol levels were completely blocked in DGKzeta-TG mouse hearts, suggesting that DGKzeta regulated PKC activity by controlling cellular diacylglycerol levels. CONCLUSIONS: These results demonstrated the first evidence that DGKzeta negatively regulated the hypertrophic signaling cascade and resultant cardiac hypertrophy in response to GPCR agonists without detectable adverse effects in in vivo hearts. PMID- 16380549 TI - Inhibition of histone deacetylation blocks cardiac hypertrophy induced by angiotensin II infusion and aortic banding. AB - BACKGROUND: A number of distinct stress signaling pathways in myocardium cause cardiac hypertrophy and heart failure. Class II histone deacetylases (HDACs) antagonize several stress-induced pathways and hypertrophy. However, cardiac hypertrophy induced by transgenic overexpression of the homeodomain only protein, HOP, can be prevented by the nonspecific HDAC inhibitors trichostatin A and valproic acid, suggesting that alternate targets that oppose class II HDAC function might exist in myocardium. We tested the effects of several HDAC inhibitors, including a class I HDAC-selective inhibitor, SK-7041, on cardiac hypertrophy induced by angiotensin II (Ang II) treatment or aortic banding (AB). METHODS AND RESULTS: Cardiac hypertrophy was induced by chronic infusion of Ang II or by AB in mice or rats and evaluated by determining the ratio of heart weight to body weight or to tibia length, cross-sectional area, or echocardiogram. Cardiac hypertrophy induced by Ang II or AB for 2 weeks was significantly reduced by simultaneous administration of trichostatin A, valproic acid, or SK-7041. Echocardiogram revealed that exaggerated left ventricular systolic dimensions were relieved by HDAC inhibitors. HDAC inhibitors partially reversed preestablished cardiac hypertrophy and improved survival of AB mice. The expressions of atrial natriuretic factor, alpha-tubulin, beta-myosin heavy chain, and interstitial fibrosis were reduced by HDAC inhibition. CONCLUSIONS: These results suggest that the predominant effect of HDAC inhibition, mainly mediated by class I HDACs, is to prevent cardiac hypertrophy in response to a broad range of agonist and stretch stimuli. PMID- 16380550 TI - Deficiency of the 5-hydroxytryptamine transporter gene leads to cardiac fibrosis and valvulopathy in mice. AB - BACKGROUND: Serotonin (5-hydroxytryptamine; 5-HT) overproduction is responsible for cardiac valvular disease in patients with carcinoid tumors. Reduced 5-HT inactivation is one proposed mechanism of the valvulopathy observed in individuals treated with the appetite suppressants fenfluramine and phentermine. One key protein limiting systemic availability of 5-HT is the 5-HT transporter (5 HTT) expressed by platelets and pulmonary vascular cells; 5-HTT is responsible for 5-HT uptake and subsequent inactivation of the amine passing through the lung. Here we investigated whether 5-HTT-deficient (5-HTT-KO) mice developed structural and/or functional cardiac abnormalities and valvulopathy. METHODS AND RESULTS: Cardiac endothelial cells expressed large amounts of 5-HTT in wild-type mice. 5-HTT deficiency appeared to be associated with marked interstitial, perivascular, and valvular fibrosis as evidenced by staining of cardiac collagen in 5-HTT-KO mice. Histological analysis provided evidence for valvulopathy characterized by valvular hyperplasia and prominent fibrosis at the attachment site and base of the leaflets. Echocardiography revealed an increase in left ventricular lumen diameter and a decrease in left ventricular diameter fractional shortening. Although 5-HT1B receptors mediated the 5-HT-induced collagen secretion by human cardiac myofibroblasts, the contribution of this receptor type to valvulopathy was ruled out because double-KO mice deficient in both 5-HTT and 5-HT1B receptors showed the same cardiac alterations as 5-HTT-KO mice. CONCLUSIONS: The present results establish a link between 5-HTT and the development of cardiac fibrosis and valvulopathy in vivo. 5-HTT-KO mice represent an especially relevant model for studying the mechanisms by which 5-HT induces valvulopathy. PMID- 16380551 TI - Sleep and exertional periodic breathing in chronic heart failure: prognostic importance and interdependence. AB - BACKGROUND: Sleep and exertional periodic breathing are proverbial in chronic heart failure (CHF), and each alone indicates poor prognosis. Whether these conditions are associated and whether excess risk may be attributed to respiratory disorders in general, rather than specifically during sleep or exercise, is unknown. METHODS AND RESULTS: We studied 133 CHF patients with left ventricular ejection fraction (LVEF) < or =40%. During 1170+/-631 days of follow up, 31 patients (23%) died. Nonsurvivors had higher New York Heart Association class, ventilatory response (ve/vco2 slope), and apnea-hypopnea index (AHI) and lower peak vo2 (all P<0.01); lower LVEF and prescription of beta-blockers, and shorter transmitral deceleration time (all P<0.05). Exertional oscillatory ventilation (EOV), established by cyclic fluctuations in minute ventilation that persisted for > or =60% of exercise duration with an amplitude > or =15% of the average resting value, was significantly more frequent in nonsurvivors (42% versus 15%, P<0.01). Multivariable analysis selected AHI (hazard ratio [HR] 5.66, 95% CI 2.3 to 19.9, P<0.01), peak vo2 (HR 0.93, 95% CI 0.90 to 0.97, P<0.01), and beta-blocker prescription (HR 0.34, 95% CI 0.13 to 0.87, P<0.05) as predictors of cardiac events. The best cutoff for AHI was >30/h. EOV was significantly related to AHI >30/h (chi2 14.6, P<0.01): 78% of EOV patients showed AHI >30/h. Multivariable analysis, including breathing disorders alone (EOV, AHI >30/h) or in combination (EOV plus AHI >30/h), selected combined disorders as the strongest predictor of events (HR 6.65, 95% CI 2.6 to 17.1, P<0.01). CONCLUSIONS: In CHF, EOV is significantly associated with AHI >30/h. Although each breathing disorder alone is linked to total mortality, their combination has a crucial prognostic burden. PMID- 16380552 TI - Comparison of electrode cooling between internal and open irrigation in radiofrequency ablation lesion depth and incidence of thrombus and steam pop. AB - BACKGROUND: Electrode cooling by circulating fluid within the electrode (closed loop) or open irrigation facilitates radiofrequency (RF) ablation. This study compared lesion parameters between closed loop and open irrigation with the use of a canine model. METHODS AND RESULTS: In 8 anesthetized dogs, the skin over the thigh muscle was incised and raised, forming a cradle superfused with heparinized blood (activated clotting time >350 seconds) at 37 degrees C. A 7F 4-mm closed loop electrode (irrigation 36 mL/min) and 7.5F 3.5-mm open irrigation electrode (irrigation 17 mL/min) were positioned perpendicular to the thigh muscle at 10 g contact weight. RF was applied (n=121) at 20 or 30 W for 60 seconds in low (0.1 m/s) or high (0.5 m/s) pulsatile blood flow. Temperatures were measured in the electrode, electrode-tissue interface, and within the tissue at 3- and 7-mm depths. After each RF, the cradle was emptied to examine the electrode and interface for thrombus. There was no difference between closed loop and open irrigation in impedance, lesion depth, or tissue temperature at 20 or 30 W. Interface temperature and electrode temperature were greater in the closed loop application. Thrombus occurred in 32 of 63 closed loop versus 0 of 58 open irrigation RF applications (P<0.05) with interface temperature > or =80 degrees C in all 32 (electrode temperature <40 degrees C in 1, 40 degrees C to 50 degrees C in 26, and >50 degrees C in 5). With closed loop, interface temperature and thrombus incidence were greater at 30 W and low blood flow. With open irrigation, interface temperature remained low (< or =71 degrees C) with no difference between 20 and 30 W or between low and high blood flow. Steam pop occurred at 20 W in 4 of 35 closed loop and 0 of 30 open irrigation and at 30 W in 15 of 28 closed loop and 4 of 28 open irrigation applications (P<0.05). CONCLUSIONS: Lower interface temperature, thrombus, and steam pop, especially in low blood flow, indicate greater interface cooling with open irrigation. PMID- 16380553 TI - Hurricane Katrina: a social and public health disaster. PMID- 16380554 TI - Appropriate use of the K function in urban environments. PMID- 16380555 TI - The diffusion of public health innovations. PMID- 16380556 TI - Daubert's menace. PMID- 16380559 TI - Did prohibition really work? Alcohol prohibition as a public health innovation. AB - The conventional view that National Prohibition failed rests upon an historically flimsy base. The successful campaign to enact National Prohibition was the fruit of a century-long temperance campaign, experience of which led prohibitionists to conclude that a nationwide ban on alcohol was the most promising of the many strategies tried thus far. A sharp rise in consumption during the early 20th century seemed to confirm the bankruptcy of alternative alcohol-control programs. The stringent prohibition imposed by the Volstead Act, however, represented a more drastic action than many Americans expected. Nevertheless, National Prohibition succeeded both in lowering consumption and in retaining political support until the onset of the Great Depression altered voters' priorities. Repeal resulted more from this contextual shift than from characteristics of the innovation itself. PMID- 16380558 TI - Growing the field of health impact assessment in the United States: an agenda for research and practice. AB - Health impact assessment (HIA) methods are used to evaluate the impact on health of policies and projects in community design, transportation planning, and other areas outside traditional public health concerns. At an October 2004 workshop, domestic and international experts explored issues associated with advancing the use of HIA methods by local health departments, planning commissions, and other decisionmakers in the United States. Workshop participants recommended conducting pilot tests of existing HIA tools, developing a database of health impacts of common projects and policies, developing resources for HIA use, building workforce capacity to conduct HIAs, and evaluating HIAs. HIA methods can influence decisionmakers to adjust policies and projects to maximize benefits and minimize harm to the public's health. PMID- 16380560 TI - Weight of the evidence or wait for the evidence? Protecting underground miners from diesel particulate matter. AB - A coalition of mine operators has used a variety of tactics to obstruct scientific inquiry and impede public health action designed to protect underground miners from diesel particulate matter. These workers are exposed to the highest level of diesel particulate matter compared with any other occupational group. This case study profiles a decade-long saga of the Methane Awareness Resource Group Diesel Coalition to impede epidemiological studies on diesel exhaust undertaken by the National Institute for Occupational Safety and Health and the National Cancer Institute, and to derail a health standard promulgated by the Mine Safety and Health Administration. The case study highlights the coalition's mastery of legislative, judicial, and executive branch operations and the reaction of policymakers. PMID- 16380561 TI - Tobacco cessation services through community health workers for Spanish-speaking populations. AB - Partnerships were established with the University of Arizona's Healthcare Partnership to train promotores--Spanish-speaking community health workers--as tobacco cessation counselors. Tobacco Free El Paso certified promotores to help identify tobacco users and offer tobacco cessation counseling services. The project certified 89 participants, of whom 95% were promotores; 88% were Hispanic/Latino, 67% were females, and 62% indicated Spanish as their primary language. Participants who completed Tecnicas Basicas, Treatment Specialist, and Dejate de ese Vicio certifications significantly increased self-confidence levels to deliver brief smoking cessation interventions (P < .05). Satisfaction scores (scale = 1-5) were also relatively high for each certification (Tecnicas Basicas, mean = 4.8; Treatment Specialist, mean = 4.7; Dejate de ese Vicio, mean = 4.6). The results suggest that promotores understood the concepts and methodologies presented. PMID- 16380562 TI - Diarrheal illness detected through syndromic surveillance after a massive power outage: New York City, August 2003. AB - OBJECTIVES: We investigated increases in diarrheal illness detected through syndromic surveillance after a power outage in New York City on August 14, 2003. METHODS: The New York City Department of Health and Mental Hygiene uses emergency department, pharmacy, and absentee data to conduct syndromic surveillance for diarrhea. We conducted a case-control investigation among patients presenting during August 16 to 18, 2003, to emergency departments that participated in syndromic surveillance. We compared risk factors for diarrheal illness ascertained through structured telephone interviews for case patients presenting with diarrheal symptoms and control patients selected from a stratified random sample of nondiarrheal patients. RESULTS: Increases in diarrhea were detected in all data streams. Of 758 patients selected for the investigation, 301 (40%) received the full interview. Among patients 13 years and older, consumption of meat (odds ratio [OR]=2.7, 95% confidence interval [CI]=1.2, 6.1) and seafood (OR=4.8; 95% CI=1.6, 14) between the power outage and symptom onset was associated with diarrheal illness. CONCLUSIONS: Diarrhea may have resulted from consumption of meat or seafood that spoiled after the power outage. Syndromic surveillance enabled prompt detection and systematic investigation of citywide illness that would otherwise have gone undetected. PMID- 16380563 TI - Dealing with an innovative industry: a look at flavored cigarettes promoted by mainstream brands. AB - Product and marketing innovation is key to the tobacco industry's success. One recent innovation was the development and marketing of flavored cigarettes as line extensions of 3 popular brands (Camel, Salem, and Kool). These products have distinctive blends and marketing as well as innovative packaging and have raised concerns in the public health community that they are targeted at youths. Several policy initiatives have aimed at banning or limiting these types of products on that basis. We describe examples of the products and their marketing and discuss their potential implications (including increased smoking experimentation, consumption, and "someday smoking"), as well as their potential impact on young adults. PMID- 16380564 TI - Breastfeeding duration and perinatal cigarette smoking in a population-based cohort. AB - OBJECTIVES: We examined the association between breastfeeding duration and maternal smoking before, during, and after pregnancy. METHODS: Data from the 2000 2001 Oregon Pregnancy Risk Assessment Monitoring System were used. Early weaning was defined as not breastfeeding at 10 weeks postpartum. RESULTS: At 10 weeks after pregnancy, 25.7% of mothers who initiated breastfeeding no longer breastfed. After controlling for confounders, quitters (mothers who quit smoking during pregnancy and maintained quit status after pregnancy) and postpartum relapsers (mothers who quit smoking during pregnancy and resumed smoking after delivery) did not have significantly higher risk for early weaning than nonsmokers. However, persistent smokers (mothers who smoked before, during, and after pregnancy) were 2.18 times more likely not to breastfeed at 10 weeks (95% confidence interval=1.52, 2.97). Women who smoked 10 or more cigarettes per day postpartum (i.e., heavy postpartum relapsers and heavy persistent smokers) were 2.3-2.4 times more likely to wean their infants before 10 weeks than were nonsmokers. CONCLUSIONS: Maternal smoking is associated with early weaning. Stopping smoking during pregnancy and decreasing the number of cigarettes smoked postpartum may increase breastfeeding duration. PMID- 16380565 TI - "Weathering" and age patterns of allostatic load scores among blacks and whites in the United States. AB - OBJECTIVES: We considered whether US Blacks experience early health deterioration, as measured across biological indicators of repeated exposure and adaptation to stressors. METHODS: Using National Health and Nutrition Examination Survey data, we examined allostatic load scores for adults aged 18-64 years. We estimated probability of a high score by age, race, gender, and poverty status and Blacks' odds of having a high score relative to Whites' odds. RESULTS: Blacks had higher scores than did Whites and had a greater probability of a high score at all ages, particularly at 35-64 years. Racial differences were not explained by poverty. Poor and nonpoor Black women had the highest and second highest probability of high allostatic load scores, respectively, and the highest excess scores compared with their male or White counterparts. CONCLUSIONS: We found evidence that racial inequalities in health exist across a range of biological systems among adults and are not explained by racial differences in poverty. The weathering effects of living in a race-conscious society may be greatest among those Blacks most likely to engage in high-effort coping. PMID- 16380567 TI - Associations of neighborhood characteristics with the location and type of food stores. AB - OBJECTIVES: We investigated associations between local food environment and neighborhood racial/ethnic and socioeconomic composition. METHODS: Poisson regression was used to examine the association of food stores and liquor stores with racial/ethnic composition and income in selected census tracts in North Carolina, Maryland, and New York. RESULTS: Predominantly minority and racially mixed neighborhoods had more than twice as many grocery stores as predominantly White neighborhoods (for predominantly Black tracts, adjusted stores per population ratio [SR]=2.7; 95% confidence interval [CI]=2.2, 3.2; and for mixed tracts, SR=2.2; 95% CI=1.9, 2.7) and half as many supermarkets (for predominantly Black tracts, SR=0.5; 95% CI=0.3, 0.7; and for mixed tracts, SR=0.7; 95% CI=0.5, 1.0, respectively). Low-income neighborhoods had 4 times as many grocery stores as the wealthiest neighborhoods (SR=4.3; 95% CI=3.6, 5.2) and half as many supermarkets (SR=0.5; 95% CI=0.3, 0.8). In general, poorer areas and non-White areas also tended to have fewer fruit and vegetable markets, bakeries, specialty stores, and natural food stores. Liquor stores were more common in poorer than in richer areas (SR=1.3; 95% CI=1.0, 1.6). CONCLUSIONS: Local food environments vary substantially by neighborhood racial/ethnic and socioeconomic composition and may contribute to disparities in health. PMID- 16380566 TI - Relative effectiveness of worker safety and health training methods. AB - OBJECTIVES: We sought to determine the relative effectiveness of different methods of worker safety and health training aimed at improving safety knowledge and performance and reducing negative outcomes (accidents, illnesses, and injuries). METHODS: Ninety-five quasi-experimental studies (n=20991) were included in the analysis. Three types of intervention methods were distinguished on the basis of learners' participation in the training process: least engaging (lecture, pamphlets, videos), moderately engaging (programmed instruction, feedback interventions), and most engaging (training in behavioral modeling, hands-on training). RESULTS: As training methods became more engaging (i.e., requiring trainees' active participation), workers demonstrated greater knowledge acquisition, and reductions were seen in accidents, illnesses, and injuries. All methods of training produced meaningful behavioral performance improvements. CONCLUSIONS: Training involving behavioral modeling, a substantial amount of practice, and dialogue is generally more effective than other methods of safety and health training. The present findings challenge the current emphasis on more passive computer-based and distance training methods within the public health workforce. PMID- 16380568 TI - Feasibility of screening adolescents for suicide risk in "real-world" high school settings. AB - OBJECTIVES: We evaluated the feasibility of a population-based approach to preventing adolescent suicide. METHODS: A total of 1323 students in 10 high schools completed the Suicide Risk Screen. Screening results, student follow-up, staff feedback, and school responses were assessed. RESULTS: Overall, 29% of the participants were rated as at risk of suicide. As a result of this overwhelming percentage, school staffs chose to discontinue the screening after 2 semesters. In further analyses, about half of the students identified were deemed at high risk on the basis of high levels of depression, suicidal ideation, or suicidal behavior. Priority rankings evidenced good construct validity on correlates such as drug use, hopelessness, and perceived family support. CONCLUSIONS: A simpler, more specific screening instrument than the Suicide Risk Screen would identify approximately 11% of urban high school youths for assessment, offering high school officials an important opportunity to identify young people at the greatest levels of need and to target scarce health resources. Our experiences from this study show that lack of feasibility testing greatly contributes to the gap between science and practice. PMID- 16380569 TI - Epidemiology and correlates of daily smoking and nicotine dependence among young adults in the United States. AB - OBJECTIVES: We describe the epidemiology of smoking behaviors in a national young adult sample and identify common and unique demographic, social, and psychological correlates of daily smoking and lifetime and current nicotine dependence by race/ethnicity. METHODS: Data are from the National Longitudinal Survey of Adolescent Health, wave III. Dependence was measured by the Revised Fagerstrom Test for Nicotine Dependence. Logistic regressions were estimated. RESULTS: Hispanic ethnicity, low education, parental and peer smoking, novelty seeking, early age of smoking onset, and pleasurable initial smoking experiences are significantly correlated with daily smoking and lifetime nicotine dependence. Depressive symptoms are uniquely associated with lifetime and current dependence. Few factors are highly associated with current dependence. Initial sensitivity to smoking has a significantly greater impact on daily smoking than on dependence. Correlates of smoking behaviors are mostly common across racial/ethnic groups, although parental and peer smoking are significant for Whites and Hispanics but not for African Americans. CONCLUSIONS: There are more common than unique correlates of each smoking stage and across racial/ethnic groups. Primary prevention and interventions addressing the factors tested could be uniform for most chronic smokers irrespective of dependence status and race/ethnicity. PMID- 16380570 TI - Socioeconomic disadvantage and periodontal disease: the Dental Atherosclerosis Risk in Communities study. AB - OBJECTIVES: We used data from the Dental Atherosclerosis Risk in Communities study to examine whether individual- and neighborhood-level socioeconomic characteristics were associated with periodontal disease. METHODS: We assessed severe periodontitis with a combination of clinical attachment loss and pocket depth measures. Marginal logistic regression modeling was used to estimate the association between individual and neighborhood socioeconomic indicators and prevalence of severe periodontitis before and after control for selected covariates. Residual intraneighborhood correlations in outcomes were taken into account in the analyses. RESULTS: Individual-level income and education were associated with severe periodontitis among Whites and African Americans, and these associations remained significant after adjustment for age, gender, recruitment center, and neighborhood socioeconomic score. Low-income Whites residing in disadvantaged neighborhoods had 1.8-fold (95% confidence interval=1.2, 2.7) higher odds of having severe periodontitis than high-income Whites residing in advantaged neighborhoods. CONCLUSIONS: Individual income and education were associated with severe periodontitis independently of neighborhood socioeconomic circumstances. Although the association between neighborhood socioeconomic status and severe periodontitis was not statistically significant, poverty and residence in a disadvantaged neighborhood were associated with higher odds of severe periodontitis among Whites. PMID- 16380571 TI - Early evidence on the effectiveness of clean indoor air legislation in New York State. AB - In July 2003, New York State implemented the Clean Indoor Air Act (CIAA) to reduce exposure to environmental tobacco smoke (ETS). In this cross-sectional study, workers (n=168) completed an interview assessing ETS exposure and provided urine for cotinine analysis. Hospitality workers recruited after implementation of the CIAA had significant reductions in ETS exposure and urine cotinine, compared with those recruited before implementation. The New York State CIAA yielded measurable reductions in ETS exposure for hospitality workers. PMID- 16380572 TI - The Environmental Protection Agency's brownfields pilot program. AB - OBJECTIVE: We studied the diffusion of the US Environmental Protection Agency's national brownfields pilot innovation to more than 300 local governments between 1993 through 2002 to determine why some local governments received grants very early in the process while other awardees received funding later. METHODS: We did an ordinal regression analysis of the characteristics of all local government award recipients, and we conducted interviews with early-award recipients. RESULTS: The first set of local government awardees had lost much of their manufacturing base, had large concentrations of economically disadvantaged minority residents, and had local capacity to compete for funding. Federal and state officials catalyzed the diffusion of the innovation by working with local governments. CONCLUSIONS: The widely praised program was diffused selectively at first and then more widely later on the basis of local need, local capacity to compete, and networks of contacts among entrepreneurs and local governments. The economic, social, political, and public health impacts must be monitored and reviewed. PMID- 16380573 TI - Cancer screening and risk factor rates among American Indians. AB - OBJECTIVES: We examined cancer screening and risk factor patterns in California using 4 different statistical tabulations of American Indian and Alaska Native (AIAN) populations. METHODS: We used the 2001 California Health Interview Survey to compare cancer screening and risk factor data across 4 different tabulation approaches. We calculated weighted prevalence estimates by gender and race/ethnicity for cancer screening and risk factors, sociodemographic characteristics, and access to care variables. We compared AIAN men and women with members of other racial groups and examined outcomes among AIAN men and women using the 4 tabulation methods. RESULTS: Although some differences were small, in general, screening and risk factor rates among American Indians/Alaska Natives were most similar to rates among Whites when the most inclusive multiracial tabulation approach was used and least similar when the more exclusive US census "single-race" approach was used. CONCLUSIONS: Racial misclassification and undercounting are among the most difficult obstacles to obtaining accurate and informative data on the AIAN population. Our analysis suggests some guidelines for overcoming these obstacles. PMID- 16380574 TI - Disparities in asthma hospitalization in Massachusetts. AB - OBJECTIVES: We examined racial disparities in asthma morbidity in Massachusetts. METHODS: We used Massachusetts case-mix data from 1994 to 2002 to screen and track individual asthma morbidity and hospitalizations, which resulted in a sample of 10145 patients who were first hospitalized for asthma between 1997 and 2000. We followed these patients for 2 years after their first hospitalization. Because asthma is widely considered a preventable cause of hospitalization, we interpreted a readmission for asthma as an indication of failed asthma management. RESULTS: We found substantial racial/ethnic disparities in readmission rates that persisted after control for comorbidities, payer type, and income. We estimated that the costs of repeat hospitalizations for asthma are in excess of one quarter of all asthma hospitalization costs. CONCLUSION: Racial/ethnic disparities in asthma readmission rates show that Massachusetts is not on the frontier of asthma treatment. PMID- 16380575 TI - Complementary and alternative medical therapy use among Chinese and Vietnamese Americans: prevalence, associated factors, and effects of patient-clinician communication. AB - OBJECTIVE: We examined the use of complementary and alternative medical (CAM) therapies among Chinese and Vietnamese Americans who had limited proficiency with the English language and explore the association between patient-clinician discussions about CAM therapy use and patient assessments of quality of care. METHODS: We surveyed Chinese and Vietnamese Americans who visited 11 community health centers in 8 major cities throughout the United States. RESULTS: Of the 4410 patients surveyed, 3258 (74%) returned completed questionnaires. Two thirds of respondents reported they had "ever used" some form of CAM therapy; however, only 7.6% of these patients had discussed their use of CAM therapies with clinicians. Among patients who had used CAM therapies during the week before their most recent visits, clinician-patient discussions about CAM therapy use were associated with better overall patient ratings of quality of care. CONCLUSION: Use of CAM therapies was common among Chinese and Vietnamese Americans who had limited proficiency with the English language. Although discussions about CAM therapy use with clinicians were uncommon, these discussions were associated with better ratings of quality of care. PMID- 16380577 TI - Physicians' perceptions of patients' social and behavioral characteristics and race disparities in treatment recommendations for men with coronary artery disease. AB - OBJECTIVES: A growing body of evidence suggests that provider decisionmaking contributes to racial/ethnic disparities in care. We examined the factors mediating the relationship between patient race/ethnicity and provider recommendations for coronary artery bypass graft surgery. METHODS: Analyses were conducted with a data set that included medical record, angiogram, and provider survey data on postangiogram encounters with patients who were categorized as appropriate candidates for coronary artery bypass graft surgery. RESULTS: Race significantly influenced physician recommendations among male, but not female, patients. Physicians' perceptions of patients' education and physical activity preferences were significant predictors of their recommendations, independent of clinical factors, appropriateness, payer, and physician characteristics. Furthermore, these variables mediated the effects of patient race on provider recommendations. CONCLUSIONS: Our findings point to the importance of research and intervention strategies addressing the ways in which providers' beliefs about patients mediate disparities in treatment. In addition, they highlight the need for discourse and consensus development on the role of social factors in clinical decisionmaking. PMID- 16380578 TI - Rapid assessment and response studies of injection drug use: knowledge gain, capacity building, and intervention development in a multisite study. AB - OBJECTIVES: We evaluated the World Health Organization's rapid assessment and response (RAR) method of assessing injection drug use and its associated health problems, focusing on knowledge gain, capacity building, and whether RAR leads to the development of interventions reducing the health effects of injection drug use. METHODS: Data were derived from RAR studies conducted in Beijing, China; Bogota, Colombia; Greater Rosario, Argentina; Hanoi, Vietnam; Kharkiv, Ukraine; Minsk, Belarus; Nairobi, Kenya; Penang, Malaysia; St. Petersburg, Russia; and Tehran, Iran. RESULTS: Substantial gains in knowledge and response capacity were reported at all of the study sites. Before RAR initiation, prevention and intervention programs had been absent or inadequate at most of the sites. The RARs resulted in many new or modified interventions; 7 sites reported 24 health related interventions that were subsequently developed and influenced by the RARs. CONCLUSIONS: RARs, which require relatively little external funding, appear to be effective in linking assessment to development of appropriate interventions. The present results add to the evidence that rapid assessment is an important public health tool. PMID- 16380579 TI - Rethinking the Hispanic paradox: death rates and life expectancy for US non Hispanic White and Hispanic populations. AB - OBJECTIVES: We examined the "Hispanic paradox," whereby persons of Hispanic origin seemed to experience lower mortality than the non-Hispanic White population. This paradox coincided with a change from the classification of deaths and population by Spanish surname to the use of Hispanic-origin questions in the census and vital statistics. METHODS: To estimate US Hispanic and non Hispanic White mortality, we applied a familiar relation between death rates for population subgroups to Hispanic and non-Hispanic White population death rates. We calculated age-specific death rates for the Hispanic population and the non Hispanic White population and computed life tables for each. Result. For Texas between 1980 (surname) and 1990 (origin), the change in Hispanic deaths in persons aged 65 years or older was only half as great as the change in population size, implying a relative omission of 15% to 20% of deaths. By a different approach, the life tables for the US Hispanic and non-Hispanic White populations pointed to a similar omission. CONCLUSIONS: There is no "Hispanic paradox." The Hispanic paradox described in past research derives from inconsistencies in counts of Hispanic-origin deaths and populations. PMID- 16380576 TI - The role of vitamin D in cancer prevention. AB - Vitamin D status differs by latitude and race, with residents of the northeastern United States and individuals with more skin pigmentation being at increased risk of deficiency. A PubMed database search yielded 63 observational studies of vitamin D status in relation to cancer risk, including 30 of colon, 13 of breast, 26 of prostate, and 7 of ovarian cancer, and several that assessed the association of vitamin D receptor genotype with cancer risk. The majority of studies found a protective relationship between sufficient vitamin D status and lower risk of cancer. The evidence suggests that efforts to improve vitamin D status, for example by vitamin D supplementation, could reduce cancer incidence and mortality at low cost, with few or no adverse effects. PMID- 16380580 TI - The cover. Children Playing Cards. PMID- 16380581 TI - Obesity's role in heart disease requires apples and pears comparison. PMID- 16380582 TI - Syphilis rates rise among men: trends for other STDs mixed. PMID- 16380583 TI - Early rheumatoid arthritis treatments weighed. PMID- 16380584 TI - Risk-treatment mismatch for heart failure. PMID- 16380585 TI - Continuing anticoagulation following venous thromboembolism. PMID- 16380586 TI - Antibiotic prescribing for lower respiratory tract infection. PMID- 16380587 TI - Long-term use of aspirin and risk of colorectal cancer. PMID- 16380588 TI - Regulation of medical marijuana. PMID- 16380590 TI - Dietary intake of antioxidants and risk of age-related macular degeneration. AB - CONTEXT: Age-related macular degeneration (AMD) is the most prevalent cause of irreversible blindness in developed countries. Recently, high-dose supplementation with beta carotene, vitamins C and E, and zinc was shown to slow the progression of AMD. OBJECTIVE: To investigate whether regular dietary intake of antioxidants is associated with a lower risk of incident AMD. DESIGN: Dietary intake was assessed at baseline in the Rotterdam Study (1990-1993) using a semiquantitative food frequency questionnaire. Incident AMD until final follow-up in 2004 was determined by grading fundus color transparencies in a masked way according to the International Classification and Grading System. SETTING: Population-based cohort of all inhabitants aged 55 years or older in a middle class suburb of Rotterdam, the Netherlands. PARTICIPANTS: Of 5836 persons at risk of AMD at baseline, 4765 had reliable dietary data and 4170 participated in the follow-up. MAIN OUTCOME MEASURE: Incident AMD, defined as soft distinct drusen with pigment alterations, indistinct or reticular drusen, geographic atrophy, or choroidal neovascularization. RESULTS: Incident AMD occurred in 560 participants after a mean follow-up of 8.0 years (range, 0.3-13.9 years). Dietary intake of both vitamin E and zinc was inversely associated with incident AMD. The hazard ratio (HR) per standard deviation increase of intake for vitamin E was 0.92 (95% confidence interval [CI], 0.84-1.00) and for zinc was 0.91 (95% CI, 0.83-0.98). An above-median intake of all 4 nutrients, beta carotene, vitamin C, vitamin E, and zinc, was associated with a 35% reduced risk (HR, 0.65; 95% CI, 0.46-0.92) of AMD. Exclusion of supplement users did not affect the results. CONCLUSION: In this study, a high dietary intake of beta carotene, vitamins C and E, and zinc was associated with a substantially reduced risk of AMD in elderly persons. PMID- 16380589 TI - Prophylactic Oral Amiodarone for the Prevention of Arrhythmias that Begin Early After Revascularization, Valve Replacement, or Repair: PAPABEAR: a randomized controlled trial. AB - CONTEXT: Atrial tachyarrhythmias after cardiac surgery are associated with adverse outcomes and increased costs. Previous trials of amiodarone prophylaxis, while promising, were relatively small and yielded conflicting results. OBJECTIVE: To determine whether a brief perioperative course of oral amiodarone is an effective and safe prophylaxis for atrial tachyarrhythmias after cardiac surgery overall and in important subgroups. DESIGN, SETTING, AND PATIENTS: Double blind randomized controlled trial of 601 patients listed for nonemergent coronary artery bypass graft (CABG) surgery and/or valve replacement/repair surgery between February 1, 1999, and September 26, 2003, at a tertiary care hospital. The patients were followed up for 1 year. INTERVENTION: Oral amiodarone (10 mg/kg daily) or placebo administered 6 days prior to surgery through 6 days after surgery (13 days). Randomization was stratified for subgroups defined by age, type of surgery, and use of preoperative beta-blockers. MAIN OUTCOME MEASURE: Incidence of atrial tachyarrhythmias lasting 5 minutes or longer that prompted therapy by the sixth postoperative day. RESULTS: Atrial tachyarrhythmias occurred in fewer amiodarone patients (48/299; 16.1%) than in placebo patients (89/302; 29.5%) overall (hazard ratio [HR], 0.52; 95% confidence interval [CI], 0.34-0.69; P<.001); in patients younger than 65 years (19 [11.2%] vs 36 [21.1%]; HR, 0.51 [95% CI, 0.28-0.94]; P = .02); in patients aged 65 years or older (28 [21.7%] vs 54 [41.2%]; HR, 0.45 [95% CI, 0.27-0.75]; P<.001); in patients who had CABG surgery only (22 [11.3%] vs 46 [23.6%]; HR, 0.45 [95% CI, 0.26-0.79]; P = .002); in patients who had valve replacement/repair surgery with or without CABG surgery (25 [23.8%] vs 44 [44.1%]; HR, 0.51 [95% CI, 0.31-0.84; P = .008); in patients who received preoperative beta-blocker therapy (27 [15.3%] vs 42 [25.0%]; HR, 0.58 [95% CI, 0.34-0.99]; P = .03); and in patients who did not receive preoperative beta-blocker therapy (20 [16.3%] vs 48 [35.8%]; HR, 0.40 [95% CI, 0.22-0.71]; P<.001), respectively. Postoperative sustained ventricular tachyarrhythmias occurred less frequently in amiodarone patients (1/299; 0.3%) than in placebo patients (8/302; 2.6%) (P = .04). Dosage reductions of blinded therapy were more common in amiodarone patients (34/299; 11.4%) than in placebo patients (16/302; 5.3%) (P = .008). There were no differences in serious postoperative complications, in-hospital mortality, or readmission to the hospital within 6 months of discharge or in 1-year mortality. CONCLUSION: Oral amiodarone prophylaxis of atrial tachyarrhythmias after cardiac surgery is effective and may be safe overall and in important patient subgroups. Clinical Trials Registration ClinicalTrials.gov Identifier: NCT00251706. PMID- 16380591 TI - Excess dosing of antiplatelet and antithrombin agents in the treatment of non-ST segment elevation acute coronary syndromes. AB - CONTEXT: Effective medical care assumes delivery of evidence-based medicines to appropriate patients with doses comparable to those studied. OBJECTIVE: To investigate dosing of unfractionated heparin (UFH), low-molecular-weight heparin (LMWH), and glycoprotein IIb/IIIa inhibitors, and the association between dosing and major outcomes. DESIGN, SETTING, AND PARTICIPANTS: A prospective observational analysis in 387 US academic and nonacademic hospitals of 30,136 patients from the CRUSADE (Can Rapid Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes With Early Implementation of the American College of Cardiology/American Heart Association Guidelines) National Quality Improvement Initiative Registry who had non-ST-segment elevation acute coronary syndromes (NSTE ACS) with chest pain and either positive electrocardiograms or cardiac biomarkers between January 1 and September 30, 2004. MAIN OUTCOME MEASURES: Excessive dosing of UFH, LMWH, and glycoprotein IIb/IIIa inhibitors and major clinical outcomes, including bleeding, in-hospital mortality, and length of stay. RESULTS: A total of 3354 patients (42%) with NSTE ACS who were administered antithrombotic agents received at least 1 initial dose outside the recommended range. An excess dose was administered to 2934 patients (32.8%) treated with UFH, 1378 (13.8%) treated with LMWH, and 2784 (26.8%) treated with glycoprotein IIb/IIIa inhibitors. Factors associated with excess dosing included older age, as well as female sex, renal insufficiency, low body weight, diabetes mellitus, and congestive heart failure. Relative to those patients not administered excess dosages, patients with excess dosages of UFH, LMWH, and glycoprotein IIb/IIIa inhibitors either tended toward or had higher risks for major bleeding (adjusted odds ratio [OR], 1.08; 95% confidence interval [CI], 0.94-1.26; OR, 1.39; 95% CI, 1.11-1.74; and OR, 1.36; 95% CI, 1.10-1.68; respectively). Bleeding increased relative to the degree of excess dose and to the number of agents administered in excess (6.6% [237/3590] if neither heparin nor glycoprotein IIb/IIIa excess vs 22.2% [93/419] if both excess). Mortality and length of stay were also higher among those patients administered excess dosing. We estimated that 15% (400/2766) of major bleeding in this population may be attributable to excess dosing. CONCLUSIONS: Patients with NSTE ACS treated in the community often receive excess doses of antithrombotic therapy. Dosing errors occur more often in vulnerable populations and predict an increased risk of major bleeding. PMID- 16380592 TI - Sibling cardiovascular disease as a risk factor for cardiovascular disease in middle-aged adults. AB - CONTEXT: While parental cardiovascular disease (CVD) doubles the risk for CVD in offspring, the extent of increased risk associated with sibling CVD is unclear. OBJECTIVE: To determine, using validated events, whether sibling CVD predicts outcome in middle-aged adults independent of other risk factors. DESIGN, SETTING, AND PARTICIPANTS: The Framingham Offspring Study, an inception cohort of the Framingham Heart Study, a prospective population-based cohort study initiated in 1948 with the offspring cohort initiated in 1971. Participants (n = 2475) were members of the offspring cohort aged 30 years or older, free of CVD, and with at least 1 sibling in the study; all were followed up for 8 years. MAIN OUTCOME MEASURES: Association of sibling CVD with 8-year personal risk for CVD using pooled logistic regression. A secondary analysis restricted to offspring with both parents in the study assessed the joint impact of parental and sibling CVD occurrence. RESULTS: Among 973 person-examinations in the sibling CVD group (mean age, 57 years) and 4506 person-examinations in the no sibling CVD group (mean age, 47 years), 329 CVD events occurred during follow-up. Baseline risk factors were more prevalent in the sibling CVD group compared with the no sibling CVD group. Sibling CVD was associated with a significantly increased risk for incident CVD (age- and sex-adjusted odds ratio [OR], 1.55; 95% confidence interval [CI], 1.19-2.03). Adjustment for risk factors did not substantially attenuate the risk (adjusted OR, 1.45; 95% CI, 1.10-1.91). In the analysis restricted to persons with both parents in the study, in models adjusting for both sibling and parental CVD, the multivariable-adjusted OR for sibling CVD (1.99; 95% CI, 1.32-3.00) exceeded that for parental CVD (1.45; 95% CI, 1.02 2.05). CONCLUSION: Using validated events, sibling CVD conferred increased risk of future CVD events above and beyond established risk factors and parental CVD in middle-aged adults. PMID- 16380593 TI - Noninvasive ventilation in acute cardiogenic pulmonary edema: systematic review and meta-analysis. AB - CONTEXT: In patients with acute cardiogenic pulmonary edema noninvasive ventilation may reduce intubation rate, but the impact on mortality and the superiority of one technique over another have not been clearly established. OBJECTIVE: To systematically review and quantitatively synthesize the short-term effect of noninvasive ventilation on major clinical outcomes. DATA SOURCES: MEDLINE and EMBASE (from inception to October 2005) and Cochrane databases (library issue 4, 2005) were searched to identify relevant randomized controlled trials and systematic reviews published from January 1, 1988, to October 31, 2005. STUDY SELECTION AND DATA EXTRACTION: Included trials were all parallel studies comparing noninvasive ventilation to conventional oxygen therapy in patients with acute pulmonary edema. Comparisons of different techniques, either continuous positive airway pressure (CPAP) or bilevel noninvasive pressure support ventilation (NIPSV), were also included. DATA SYNTHESIS: Fifteen trials were selected. Overall, noninvasive ventilation significantly reduced the mortality rate by nearly 45% compared with conventional therapy (risk ratio [RR], 0.55; 95% confidence interval [CI], 0.40-0.78; P = .72 for heterogeneity). The results were significant for CPAP (RR, 0.53; 95% CI, 0.35-0.81; P = .44 for heterogeneity) but not for NIPSV (RR, 0.60; 95% CI, 0.34-1.05; P = .76 for heterogeneity), although there were fewer studies in the latter. Both modalities showed a significant decrease in the "need to intubate" rate compared with conventional therapy: CPAP (RR, 0.40; 95% CI, 0.27-0.58; P = .21 for heterogeneity), NIPSV (RR, 0.48; 95% CI, 0.30-0.76; P = .24 for heterogeneity), and together (RR, 0.43; 95% CI, 0.32-0.57; P = .20 for heterogeneity). There were no differences in intubation or mortality rates in the analysis of studies comparing the 2 techniques. CONCLUSIONS: Noninvasive ventilation reduces the need for intubation and mortality in patients with acute cardiogenic pulmonary edema. Although the level of evidence is higher for CPAP, there are no significant differences in clinical outcomes when comparing CPAP vs NIPSV. PMID- 16380594 TI - Adrenoleukodystrophy: new approaches to a neurodegenerative disease. AB - X-linked adrenoleukodystrophy (X-ALD), which was first described in 1923, was viewed until 1976 as a rare and inexorably fatal neurodegenerative disorder that affected boys. The genetic defect and biochemical abnormalities have now been defined. Ongoing research has resulted in new findings: (1) there is a wide range of phenotypic expression. At least half of patients with X-ALD are adults with somewhat milder manifestations, and women who are carriers may become symptomatic. X-ALD is often misdiagnosed as attention-deficit/hyperactivity disorder in boys and as multiple sclerosis in men and women, and is not an uncommon cause of Addison disease; (2) the incidence of X-ALD, estimated to be 1:17,000 in all ethnic groups, approximates that of phenylketonuria; (3) noninvasive and presymptomatic diagnosis and prenatal diagnosis are available; family screening and genetic counseling are key to disease prevention; and (4) new therapies, applied early, show promise. Neonatal screening is likely to become available, and a wider awareness of X-ALD and its various modes of presentation permit new proactive approaches to this distressing disorder. PMID- 16380595 TI - The 100-year anniversary of the description of the frozen section procedure. PMID- 16380596 TI - Technology licensing: lessons from the US experience. PMID- 16380597 TI - Prophylaxis against postoperative atrial fibrillation: current progress and future directions. PMID- 16380598 TI - JAMA patient page. Frozen section biopsy. PMID- 16380600 TI - Multiple populations of T lymphocytes are distinguished by the level of CD4 and CD8 coexpression and require individual consideration. PMID- 16380601 TI - A study of CD33 (SIGLEC-3) antigen expression and function on activated human T and NK cells: two isoforms of CD33 are generated by alternative splicing. AB - The expression of CD33, a restricted leukocyte antigen considered specific for myeloid lineage, has been studied extensively on lymphoid cells. We demonstrated that wide subsets of mitogen- or alloantigen-activated human T and natural killer (NK) cells express CD33 at protein and nucleic acid levels. CD33+ and CD33- T and NK cell populations showed identical surface expression of activation markers such as CD25, CD28, CD38, CD45RO, or CD95. Myeloid and lymphoid CD33 cDNA were identical. However, lymphoid CD33 protein had lower molecular weight, suggesting cell type-specific, post-translational modifications. Additionally, reverse transcriptase-polymerase chain reaction and Northern blot analysis showed an unknown CD33 isoform (CD33m) expressed on all CD33+ cell lines or T cell clones tested. CD33m was identical to CD33 (CD33M) in the signal peptide, the immunoglobulin (Ig) domain C2, the transmembrane, and the cytoplasmic regions but lacked the extracellular ligand-binding variable Ig-like domain encoded by the second exon. CD33m mRNA was mostly detected on NKL and myeloid cell lines but poorly expressed on B cell lines and T lymphocytes. The CD33m extracellular portion was successfully expressed as a soluble fusion protein on transfected human cells, suggesting a functional role on cell membranes. Cross-linking of CD33 diminished the cytotoxic activity of NKL cells against K562 and P815 target cells, working as an inhibitory receptor on NK cells. These data demonstrate that CD33 expression is not restricted to the myeloid lineage and could exist as two different splicing variants, which could play an important role in the regulation of human lymphoid and myeloid cells. PMID- 16380602 TI - Peroxisome proliferator-activated receptor gamma contributes to T lymphocyte apoptosis during sepsis. AB - In the last two decades, extensive research failed to significantly improve the outcome of patients with sepsis. In part, this drawback is based on a gap in our knowledge about molecular mechanisms understanding the pathogenesis of sepsis. During sepsis, T cells are usually depleted. Recent studies in mice and human cells suggested a role of the peroxisome proliferator-activated receptor gamma (PPARgamma) in provoking apoptosis in activated T lymphocytes. Therefore, we studied whether expression/activation of PPARgamma might contribute to T cell death during sepsis. We observed PPARgamma up-regulation in T cells of septic patients. In contrast to controls, PPARgamma expressing cells from septic patients responded with apoptosis when exposed to PPARgamma agonists. Cell demise was attenuated by SR-202, a synthetic PPARgamma antagonist, and specificity was further verified by excluding a proapoptotic response to a PPARalpha agonist. We propose that up-regulation of PPARgamma sensitizes T cells of septic patients to undergo apoptosis. PPARgamma activation in T cells requires an exogenous PPARgamma agonist, which we identified in sera of septic patients. Septic sera were used to study reporter gene expression containing a PPAR-responsive element. We conclude that PPARgamma plays a significant role in T cell apoptosis, contributing to lymphocyte loss in sepsis. Thus, inhibition of PPARgamma may turn out to be beneficial for patients suffering from lymphopenia during sepsis. PMID- 16380603 TI - Patient page. Not all dementia is Alzheimer: dementia with Lewy bodies. PMID- 16380604 TI - Atypical territorial infarction in moyamoya disease. PMID- 16380605 TI - Nature throws curveballs: APOE and nonlinear decline in Alzheimer disease. PMID- 16380606 TI - Acute transverse myelitis: is the "idiopathic" form vanishing? PMID- 16380607 TI - Determinants of rapid disease progression in ALS. PMID- 16380608 TI - APOE alleles predict the rate of cognitive decline in Alzheimer disease: a nonlinear model. AB - BACKGROUND: The APOE genotype predicts the age at onset of Alzheimer disease (AD) and neuropathologic progression. However, studies relating APOE alleles to the rate of cognitive decline have been inconclusive. This may stem from their use of linear statistical analyses. OBJECTIVE: To model relations of APOE alleles to the rate of cognitive decline in AD, nonlinearly. METHODS: Serial measures of cognitive ability were obtained using the cognitive scale of the Cambridge Examination for Mental Disorders of the Elderly in 218 patients with AD. The relations of these serial scores to APOE alleles were tested using nonlinear and linear mixed-effects models. RESULTS: In the non-linear model, possession of an APOE epsilon4 allele related to earlier and faster cognitive decline, but possession of an APOE epsilon2 related to slower decline. Patients homozygous for APOE epsilon4 showed faster cognitive decline than heterozygotes. The linear model was less sensitive and did not detect differences between APOE epsilon4 homo- and heterozygotes. CONCLUSIONS: APOE genotype strongly predicts the rate of cognitive decline in Alzheimer disease. The decline shows a dose-response relation with the APOE epsilon4 allele, but the APOE epsilon2 allele is protective. The nonlinear model yielded larger estimates of the maximal rate of decline than the linear. PMID- 16380609 TI - Prodromal cognitive signs of dementia in 85-year-olds using four sources of information. AB - OBJECTIVE: To evaluate the utility of assessing four cognitive domains obtained from four information sources to identify individuals at risk for developing dementia, Alzheimer disease (AD), and vascular dementia (VaD). METHODS: A representative population sample of nondemented 85-year-olds (n = 313) from Gothenburg, Sweden, was examined regarding memory, language, and visuospatial and executive functions, using self- and key informant reports and neuropsychiatric and psychometric examinations. The sample was followed 3 years for incident dementia, AD, or possible VaD. RESULTS: All cases of dementia, AD, and VaD were preceded by low performance in most domains irrespective of information source. Isolated low memory performance or low cognitive performance with unimpaired memory did not predict dementia. Positive predictive values increased with number of domains affected. Self- and key informant reports were less useful for predicting dementia than neuropsychiatric and psychometric examinations. The best positive predictive value (88%) was for low cognitive performance in all domains using neuropsychiatric and psychometric examinations; however, sensitivity was only 18%. CONCLUSIONS: Although memory impairment was necessary to predict dementia, it was not sufficient. Other cognitive domains needed to be affected. Relying on self-reports or key informants for early detection of dementia excluded a large group at risk. In addition, vascular dementia appeared to have a prodromal stage. PMID- 16380610 TI - Parkinsonian signs in subjects with mild cognitive impairment. AB - BACKGROUND: Parkinsonian signs such as gait disturbance, rigidity, bradykinesia, and tremor are common among individuals with dementia and are associated with negative outcomes, but little is known about parkinsonian signs among individuals with mild cognitive impairment (MCI). OBJECTIVE: To examine the extent to which MCI is associated with parkinsonian signs and the relation between cognitive abilities and parkinsonism among individuals with MCI. METHODS: Participants included 835 individuals from the Rush Memory and Aging Project, a clinical pathologic study of common chronic conditions of old age. All participants underwent detailed clinical evaluations which included assessments of parkinsonian signs and cognitive function, and linear regression models were used to examine the associations of MCI and parkinsonism. RESULTS: In a series of analyses controlled for age, sex, and education, individuals with MCI exhibited significantly more parkinsonism than individuals without cognitive impairment, particularly gait disturbance, bradykinesia, and rigidity. Among individuals with MCI, lower levels of cognitive function, particularly in perceptual speed, were associated with higher levels of parkinsonism; when classified according to MCI subtype, individuals with amnestic vs non-amnestic MCI differed from each other on only one parkinsonian sign, with non-amnestic MCI showing more gait disturbance. Because vascular factors can contribute to cognitive impairment and parkinsonian signs, the authors repeated the core analyses including terms for vascular risk factors and vascular disease and the associations between MCI and parkinsonism persisted. CONCLUSIONS: Mild cognitive impairment (MCI) is accompanied by parkinsonian signs, which are related to the severity and type of cognitive impairment. The association between MCI and parkinsonism is not explained by vascular risk factors or vascular disease. PMID- 16380611 TI - Orolingual angioedema associated with ACE inhibitor use after rtPA treatment of acute stroke. PMID- 16380612 TI - Cerebral microbleeds are common in ischemic stroke but rare in TIA. AB - BACKGROUND: In patients with stroke, gradient-echo MRI commonly detects microbleeds, indicating small artery disease with increased risk of macroscopic intracranial bleeding. Antithrombotic treatments are frequently prescribed after TIA and stroke, but there have been no previous studies of microbleeds in TIA. Because microbleeds may predict the hemorrhagic risk of antithrombotic treatments, we studied the prevalence of microbleeds, risk factors, and pathophysiologic mechanisms in patients with ischemic stroke and TIA. METHODS: One hundred twenty-nine consecutive patients with ischemic stroke or TIA were studied with MRI including T2, fluid-attenuated inversion recovery, and gradient echo MRI sequences. Blinded observers counted microbleeds and graded white matter T2 hyperintensities throughout the brain. TIA patients with previous ischemic stroke were excluded. RESULTS: Sixty-seven percent of patients had ischemic stroke; 33% had TIA. Microbleeds were found in 23% of ischemic stroke patients but only 2% of TIA patients (p < 0.001). There were no significant differences in conventional risk factors or the severity of white matter disease on T2 MRI between stroke and TIA patients. Patients with microbleeds were more often hypertensive (81 vs 59%; p = 0.04) and had more severe MRI white matter disease on T2 MRI (p = 0.003). CONCLUSIONS: Microbleeds are common in ischemic stroke but rare in TIA, an observation not explained by differences in vascular risk factors or severity of white matter disease seen on T2 MRI. This finding has implications for the safety of antithrombotic therapy and clinical trial design in the two groups. Microbleeds may also be a new marker for severe microvascular pathology with increased risk of permanent cerebral infarction. PMID- 16380613 TI - The natural history of primary progressive MS in British Columbia, Canada. AB - BACKGROUND: Primary progressive multiple sclerosis (PPMS) has a distinct clinical phenotype and has historically been understudied with few longitudinal natural history studies spanning a reasonable time period. The authors examined patient characteristics, disease progression, and associated risk factors in the PP population of British Columbia, Canada. METHOD: The authors report on the PP population from an upcoming publication of the natural history study of definite MS in British Columbia, Canada. The main outcome was sustained progression to Expanded Disability Status Scale (EDSS) 6, secondary outcome time to EDSS 8. Risk factors for progression included sex, onset age, onset symptoms, and for time to EDSS 8, time to EDSS 6. RESULTS: Of the 2,837 patients with definite MS in the original study, 352 (12.4%) had PPMS. Mean disease duration was 17.2 years (0.3 to 49.6 years), mean onset age was 40.1 years (SD 11.5), and 53% (187) were female. One-quarter of the population had reached EDSS 6 after 7.3 years from onset, yet another 25% still did not require a cane after 25 years. Sex, onset age, and onset symptoms did not predict progression (p > 0.05). A shorter time to EDSS 6 predicted a shorter time to EDSS 8 (p < 0.0005). DISCUSSION: Progression of disability was slower than found in previous primary progressive multiple sclerosis natural history studies. However, considerable variation existed, with few predictors, other than "sooner to cane, sooner to wheelchair." PMID- 16380614 TI - Macrophage clustering as a diagnostic marker in sural nerve biopsies of patients with CIDP. AB - BACKGROUND: In adult patients with a slowly progressive demyelinating neuropathy, it may be difficult to distinguish between a hereditary neuropathy and chronic inflammatory demyelinating polyneuropathy (CIDP). The authors previously observed clustering of macrophages around endoneurial blood vessels in sural nerve biopsies from patients with CIDP. OBJECTIVES: To quantitate macrophage clustering around endoneurial blood vessels in CIDP vs hereditary neuropathies. METHODS: The authors studied 21 patients with CIDP, 18 patients with hereditary neuropathies, and 5 normal sural nerves. Numbers of macrophages, T-cells, and blood vessels were counted after immunohistochemical staining. The presence of three or more macrophages around one blood vessel was defined as a cluster. In a subsequent validation analysis, 65 stored biopsy specimens obtained from patients with a chronic neuropathy were re-evaluated for perivascular macrophage clustering according to criteria derived from the quantitative analysis of the first 221 biopsies in a blinded fashion. RESULTS: The percentage of endoneurial vessels with macrophage clusters was higher in CIDP than in hereditary neuropathies (CIDP median = 9.4, range 0 to 48; hereditary NP median = 0, range 0 to 7.7; p < 0.001). The evaluation of the 65 further biopsies showed that the presence of one perivascular macrophage cluster per fascicle proved to be a valid criterion to differentiate between inflammatory and other forms of neuropathy (chi2 test p = 0.0000025, sensitivity 75%, specificity 72%). CONCLUSION: The presence of clusters of macrophages around endoneurial vessels in sural nerve biopsies may serve as a useful additional marker for establishing the pathologic diagnosis of chronic inflammatory demyelinating polyneuropathy (CIDP). PMID- 16380615 TI - Minicore myopathy with ophthalmoplegia caused by mutations in the ryanodine receptor type 1 gene. AB - BACKGROUND: Minicore myopathy (multi-minicore disease [MmD]) is a congenital myopathy characterized by multifocal areas with loss of oxidative activity on muscle biopsy. MmD is clinically heterogeneous and distinct phenotypes have been associated with recessive mutations in either the selenoprotein N (SEPN1) or the skeletal muscle ryanodine receptor (RYR1) gene, also implicated in central core disease and malignant hyperthermia. External ophthalmoplegia is an additional finding in a subset of patients with MmD. OBJECTIVE: To clinically and genetically examine families with MmD and external ophthalmoplegia. METHODS: The authors investigated 11 affected individuals from 5 unrelated families. Clinical, histopathologic, and imaging studies were performed and RYR1 haplotyping and mutational analysis were carried out. RESULTS: All patients had multiple cores involving the entire fiber diameter on longitudinal sections. Weakness and wasting in the shoulder girdle, scoliosis, moderate respiratory impairment, and feeding difficulties were prominent. In contrast to SEPN1-related myopathies, soleus was more severely affected than gastrocnemius on muscle MRI. Haplotyping suggested linkage to the RYR1 locus in informative families and mutational screening revealed four novel RYR1 mutations in three unrelated families; in addition, functional haploinsufficiency was found in one allele of two recessive cases. CONCLUSION: These findings expand the phenotypic spectrum associated with mutations in the skeletal muscle ryanodine receptor (RYR1) gene. Recessive mutations of domains commonly affected in malignant hyperthermia appear to be particularly prevalent in multi-minicore disease with external ophthalmoplegia and might suggest a different pathomechanism from that involved in central core disease. PMID- 16380616 TI - A mutation in myotilin causes spheroid body myopathy. AB - BACKGROUND: Spheroid body myopathy (SBM) is a rare, autosomal dominant, neuromuscular disorder, which has only been previously reported in a single large kindred. Identification of the mutated gene in this disorder may provide insight regarding abnormal neuromuscular function. METHODS: The authors completed a detailed clinical evaluation on an extensive kindred diagnosed with SBM. Genome wide linkage analysis was performed to localize the disease gene to a specific chromosomal region. Further marker genotyping and screening of a positional, functional candidate gene were completed to detect the disease-causing mutation. Pathologic analysis of muscle biopsy was performed on three individuals. Biochemical studies were performed on one muscle biopsy specimen from an affected individual. RESULTS: Linkage to chromosome 5q23-5q31 was detected with a lod score of 2.9. Genotyping of additional markers in a larger sample of family members produced a maximum lod score of 6.1 and narrowed the critical interval to 12.2 cM. Screening of the candidate gene titin immunoglobulin domain protein (TTID, also known as MYOT) detected a cytosine-to-thymine mutation in exon 2 of all clinically affected family members. Similar pathologic changes were present in all muscle biopsy specimens. Immunohistologic and biochemical analysis revealed that the TTID protein, also known as myotilin, is a component of the insoluble protein aggregate. CONCLUSIONS: A novel mutation in the TTID gene results in the clinical and pathologic phenotype termed "spheroid body myopathy." Mutations in this gene also cause limb-girdle muscular dystrophy 1A and are associated with myofibrillar myopathy. PMID- 16380618 TI - Idiopathic acute transverse myelitis: application of the recent diagnostic criteria. AB - Despite an extensive diagnostic workup, some cases of acute transverse myelitis (ATM) remain of unknown etiology and have been referred to as "idiopathic" by the Transverse Myelitis Consortium group. In a retrospective study of 288 patients with ATM, 45 cases (15.6%) met the criteria for idiopathic ATM. The patients formed a relatively homogeneous group in terms of clinical and MRI data, but the prognosis was highly variable. PMID- 16380617 TI - Atomoxetine treatment in children and adolescents with ADHD and comorbid tic disorders. AB - OBJECTIVE: To test the hypothesis that atomoxetine does not significantly worsen tic severity relative to placebo in children and adolescents with attention deficit/hyperactivity disorder (ADHD) and comorbid tic disorders. METHODS: Study subjects were 7 to 17 years old, met Diagnostic and Statistical Manual of Mental Disorders-IV criteria for ADHD, and had concurrent Tourette syndrome or chronic motor tic disorder. Patients were randomly assigned to double-blind treatment with placebo (n = 72) or atomoxetine (0.5 to 1.5 mg/kg/day, n = 76) for up to 18 weeks. RESULTS: Atomoxetine treatment was associated with greater reduction of tic severity at endpoint relative to placebo, approaching significance on the Yale Global Tic Severity Scale total score (-5.5 +/- 6.9 vs -3.0 +/- 8.7, p = 0.063) and Tic Symptom Self-Report total score (-4.7 +/- 6.5 vs -2.9 +/- 5.2, p = 0.095) and achieving significance on the Clinical Global Impressions (CGI) tic/neurologic severity scale score (-0.7 +/- 1.2 vs -0.1 +/- 1.0, p = 0.002). Atomoxetine patients also showed greater improvement on the ADHD Rating Scale total score (-10.9 +/- 10.9 vs -4.9 +/- 10.3, p < 0.001) and CGI severity of ADHD/psychiatric symptoms scale score (-0.8 +/- 1.1 vs -0.3 +/- 1.0, p = 0.015). Discontinuation rates were not significantly different between treatment groups. Atomoxetine patients had greater increases in heart rate and decreases of body weight, and rates of treatment-emergent decreased appetite and nausea were higher. No other clinically relevant treatment differences were seen in any other vital sign, adverse event, or electrocardiographic or laboratory measures. CONCLUSIONS: Atomoxetine did not exacerbate tic symptoms. Rather, there was some evidence of reduction in tic severity with a significant reduction of attention deficit/hyperactivity disorder symptoms. Atomoxetine treatment appeared safe and well tolerated. PMID- 16380619 TI - Elevated IL-6 and TNF-alpha levels in patients with ALS: inflammation or hypoxia? AB - Abnormal levels of interleukin (IL)-6 were described in patients with ALS, related to an inflammatory process. The authors compared IL-6 and tumor necrosis factor alpha (TNF-alpha) levels in CSF and sera from 10 hypoxemics and 10 normoxemics patients with ALS to those of 10 hypoxemics and 10 normoxemics neurologic controls. The same pattern exists in patients with ALS and controls: the highest levels are found in hypoxic conditions and undetectable levels are found in normoxemic conditions. Elevated IL-6 levels in ALS could correspond to a normal response to hypoxemia. PMID- 16380620 TI - Pregnancy, delivery, and birth outcome in women with multiple sclerosis. AB - Using data from the compulsory Medical Birth Registry of Norway, the authors investigated the effect of maternal multiple sclerosis (MS) on pregnancy, delivery, and birth outcome in 649 births by MS mothers and 2.1 million control births. The mothers with MS had a higher proportion of neonates small for gestational age and also more frequent induction and operative interventions during delivery. PMID- 16380621 TI - Immune response to influenza vaccine is maintained in patients with multiple sclerosis receiving interferon beta-1a. AB - Immunologic response was assessed prospectively using influenza vaccine in 86 patients with multiple sclerosis (MS) who were taking interferon beta-1a and 77 patients who were not taking interferon. Blood samples were assayed for hemagglutination inhibition (HI) titers 0, 21, and 28 days after immunization. The two groups were similar in the proportion of patients achieving an HI titer of 40 or greater, the prespecified primary end point and on all secondary indicators of immune response. PMID- 16380622 TI - Dopamine transporter imaging study in parkinsonism occurring in fragile X premutation carriers. AB - The authors studied four patients with parkinsonism carrying the fragile X premutation using SPECT with ([23)I]FP-CIT. They found evidence of preserved presynaptic nigrostriatal function, suggesting that parkinsonism in the X fragile premutation might be related to postsynaptic dopaminergic changes or different neurotransmitter alterations. PMID- 16380623 TI - Gender differences in handedness and speech lateralization related to early neurologic insults. AB - Left-handedness is a normal variant but also may result from early-life brain insults. Speech typically resides in the left cerebral hemisphere in normal subjects. In 170 subjects with past brain injuries, bilateral or right hemisphere speech lateralization, determined by the intracarotid amobarbital procedure, was more frequent in females (19 vs 5, p = 0.003) as was left-handedness (26 vs 10, p = 0.004). This could be attributed to greater plasticity or functional symmetry in females. PMID- 16380624 TI - Hyponatremia from oxcarbazepine and carbamazepine. AB - The authors examined sodium concentrations from 97 oxcarbazepine-treated (OXC) and 451 carbamazepine-treated (CBZ) patients with epilepsy using cross-section and follow-up studies. The frequency of hyponatremia (Na+ < or = 134 mEq/L) was 29.9% among OXC-treated patients and 13.5% among CBZ-treated patients (p < 0.0001). Hyponatremia (Na+ < or = 128 mEq/L) was severe: 12.4% of OXC-treated patients and 2.8% of CBZ-treated patients (p < 0.001). Advanced age was a risk factor for hyponatremia. Hyponatremia, once present, persisted in both groups. PMID- 16380625 TI - Preictal headache in partial epilepsy. AB - The authors studied clinical characteristics in 11 patients with intractable focal epilepsy and preictal headache (PIHA) using a standardized interview. Headache was frontotemporal, ipsilateral to the focus, in nine patients with temporal lobe epilepsy (TLE) and contralateral in one with TLE and in one with frontal seizures. Migrainous features were found in four. After surgery, all seven seizure-free patients and two with rare seizures were free of PIHA. It may be a useful lateralizing sign in patients with TLE. PMID- 16380626 TI - CCM3 mutations are uncommon in cerebral cavernous malformations. AB - Cerebral cavernous malformations (CCMs) are characterized by abnormally enlarged capillary cavities without intervening brain parenchyma. Mutations in the gene PDCD10 have been found in CCM families linked to the CCM3 locus. The authors screened this gene in 15 families that did not have a CCM1 or CCM2 mutation. Only two novel mutations were found, suggesting that mutations in this gene may only account for a small percentage of CCM familial cases. PMID- 16380627 TI - Dystrophin analysis in carriers of Duchenne and Becker muscular dystrophy. AB - Associations between clinical phenotype (muscle weakness, dilated cardiomyopathy) and dystrophin abnormalities in muscle tissue among definite carriers of Duchenne (DMD) and Becker muscular dystrophy (BMD) were investigated. No associations between dystrophin abnormalities and clinical variables in DMD/BMD carriers were found. Because 26% of nonmanifesting carriers have dystrophin-negative fibers, this might be used in suspected DMD/BMD carriers in whom DNA analysis fails to give an answer about their carrier risk. PMID- 16380629 TI - MuSK Ab described in seropositive MG sera found to be Ab to alkaline phosphatase. PMID- 16380628 TI - Wernicke encephalopathy after bariatric surgery: losing more than just weight. PMID- 16380630 TI - A trochlear stroke. PMID- 16380631 TI - Spontaneous bilateral carotid artery dissection and posterior reversible encephalopathy syndrome. PMID- 16380632 TI - The therapeutic paradox in the diagnosis of tuberculous meningitis. PMID- 16380633 TI - Memoir: never saw it coming. PMID- 16380634 TI - Modafinil for fatigue in MS: a randomized placebo-controlled double-blind study. PMID- 16380635 TI - A variant in the HS1-BP3 gene is associated with familial essential tremor. PMID- 16380636 TI - Mitoxantrone treatment of multiple sclerosis: safety considerations. PMID- 16380637 TI - Personal history: the pipes of Pan. PMID- 16380638 TI - Overexpression of estrogen receptor alpha increases hepatic cholesterogenesis, leading to biliary hypersecretion in mice. AB - We explored whether there is an "estrogen-ERalpha-SREBP-2" (for estrogen-estrogen receptor subtype alpha-sterol-regulatory element binding protein-2) pathway for regulating hepatic cholesterol biosynthesis in ovariectomized AKR mice treated with 17beta-estradial (E2) at 6 microg/day or E2 plus the antiestrogenic agent ICI 182,780 at 125 microg/day and on chow or fed a high-cholesterol (1%) diet for 14 days. To monitor changes in cholesterol biosynthesis and newly synthesized cholesterol secreted into bile, incorporation into digitonin-precipitable sterols in mice treated with 25 mCi of [3H]water was measured in extracts of liver and extrahepatic organs 1 h later and in hepatic biles 6 h later. ERalpha upregulated SREBP-2, with resulting activation of SREBP-2-responsive genes in the cholesterol biosynthetic pathway. The E2-treated mice continued to synthesize cholesterol in spite of its excess availability from high dietary cholesterol, which reflects a loss in controlling the negative feedback regulation of cholesterol synthesis. These alterations augmented biliary cholesterol secretion and enhanced the lithogenicity of bile. However, these lithogenic effects of E2 were fully blocked by ICI 182,780. We conclude that during estrogen treatment, more newly synthesized cholesterol determined by the estrogen-ERalpha-SREBP-2 pathway is secreted into bile, leading to biliary cholesterol hypersecretion. These studies provide insights into therapeutic approaches to cholesterol gallstones in high risk subjects, especially those exposed to high levels of estrogen. PMID- 16380639 TI - Physiologic interactions between macrophages and Leydig cells. AB - The purpose of this minireview is to present information concerning the morphologic and functional relationship between testicular macrophages and Leydig cells. Although data concerning the negative influence of macrophage-derived products on testicular Leydig cells exist, this review is focused on the stimulatory influences thought to be involved in the physiologic interactions between these two diverse cell types. PMID- 16380640 TI - Adult stem cells for cardiac repair: a choice between skeletal myoblasts and bone marrow stem cells. AB - The real promise of a stem cell-based approach for cardiac regeneration and repair lies in the promotion of myogenesis and angiogenesis at the site of the cell graft to achieve both structural and functional benefits. Despite all of the progress and promise in this field, many unanswered questions remain; the answers to these questions will provide the much-needed breakthrough to harness the real benefits of cell therapy for the heart in the clinical perspective. One of the major issues is the choice of donor cell type for transplantation. Multiple cell types with varying potentials have been assessed for their ability to repopulate the infarcted myocardium; however, only the adult stem cells, that is, skeletal myoblasts (SkM) and bone marrow-derived stem cells (BMC), have been translated from the laboratory bench to clinical use. Which of these two cell types will provide the best option for clinical application in heart cell therapy remains arguable. With results pouring in from the long-term follow-ups of previously conducted phase I clinical studies, and with the onset of phase II clinical trials involving larger population of patients, transplantation of stem cells as a sole therapy without an adjunct conventional revascularization procedure will provide a deeper insight into the effectiveness of this approach. The present article discusses the pros and cons of using SkM and BMC individually or in combination for cardiac repair, and critically analyzes the progress made with each cell type. PMID- 16380641 TI - Role of matrix metalloproteinase-7 (matrilysin) in human cancer invasion, apoptosis, growth, and angiogenesis. AB - Matrix metalloproteinase (MMP)-7, also known as matrilysin, is a "minimal domain MMP" that exhibits proteolytic activity against components of the extracellular matrix (ECM). Matrilysin is frequently overexpressed in human cancer tissues and is associated with cancer progression. Tumorigenesis is a multistep process involving cell growth, invasion, metastasis, and angiogenesis. Matrilysin has been shown to play important roles not only in degradation of ECM proteins, but also in the regulation of several biochemical processes such as activation, degradation, and shedding of non-ECM proteins. This minire-view provides a summary of the current literature on the roles of matrilysin in tumorigenesis with a focus on the roles of modifications of non-ECM proteins by matrilysin and other related MMPs in tumorigenesis. Proteolysis of insulin-like growth factor binding protein by matrilysin results in increased bioavailability of insulin like growth factors and enhanced cellular proliferation. Matrilysin has also been implicated in the ectodomain shedding of several cell surface molecules. Heparin binding epidermal growth factor precursor (proHB-EGF) is cleaved by matrilysin into mature HB-EGF, which promotes cellular proliferation. Membrane-bound Fas ligand (FasL) is cleaved into soluble FasL, which increases apoptosis of cells adjacent to tumor cells. E-cadherin is converted to soluble E-cadherin to promote invasion. Tumor necrosis factor (TNF)-alpha precursor is cleaved to release soluble TNF-alpha to increase apoptosis. We propose that these matrilysin mediated pathways provide the necessary and logical mechanisms to promote cancer progression. PMID- 16380642 TI - Measuring rotations of a few cross-bridges in skeletal muscle. AB - The ability to measure properties of a single cross-bridge in working muscle is important because it avoids averaging the signal from a large number of molecules and because it probes cross-bridges in their native crowded environment. Because the concentration of myosin in muscle is large, observing the kinetics of a single myosin molecule requires that the signal be collected from small volumes. The introduction of small observational volumes defined by diffraction-limited laser beams and confocal detection has made it possible to limit the observational volume to a femtoliter (10(-15) liter). By restraining labeling to 1 fluorophore per 100 myosin molecules, we were able to follow the kinetics of approximately 400 cross-bridges. To reduce this number further, we used two photon (2P) microscopy. The focal plane in which the laser power density was high enough to produce 2P absorption was thinner than in confocal microscopy. Using 2P microscopy, we were able to observe approximately 200 cross-bridges during contraction. The novel method of confocal total internal reflection (CTIR) provides a method to reduce the observational volume even further, to approximately 1 attoliter (10(-18) liter), and to measure fluorescence with a high signal-to-noise (S/N) ratio. In this method, the observational volume is made shallow by illuminating the sample with an evanescent field produced by total internal reflection (TIR) of the incident laser beam. To guarantee the small lateral dimensions of the observational volume, a confocal aperture is inserted in the conjugate-image plane of the objective. With a 3.5-mum confocal aperture, we achieved a volume of 1.5 attoliter. Association-dissociation of the myosin head was probed with rhodamine attached at cys707 of the heavy chain of myosin. Signal was contributed by one to five fluorescent myosin molecules. Fluorescence decayed in a series of discrete steps, corresponding to bleaching of individual molecules of rhodamine. The S/N ratio was sufficiently large to make statistically significant comparisons from rigor and contracting myofibrils. PMID- 16380643 TI - Mesenchymal stem cells and the treatment of cardiac disease. AB - The ischemia-induced death of cardiomyocytes results in scar formation and reduced contractility of the ventricle. Several preclinical and clinical studies have supported the notion that cell therapy may be used for cardiac regeneration. Most attempts for cardiomyoplasty have considered the bone marrow as the source of the "repair stem cell(s)," assuming that the hematopoietic stem cell can do the work. However, bone marrow is also the residence of other progenitor cells, including mesenchymal stem cells (MSCs). Since 1995 it has been known that under in vitro conditions, MSCs differentiate into cells exhibiting features of cardiomyocytes. This pioneer work was followed by many preclinical studies that revealed that ex vivo expanded, bone marrow-derived MSCs may represent another option for cardiac regeneration. In this work, we review evidence and new prospects that support the use of MSCs in cardiomyoplasty. PMID- 16380644 TI - Influence of a protein concentrate from Amaranthus cruentus seeds on lipid metabolism. AB - It is widely known that elevated cholesterol and triglycerides levels favor the development of heart disease. In this paper we studied the effect of a protein concentrate from Amaranthus cruentus (Ac) on the lipid content in serum and liver tissue of male Wistar rats. The animals were separated into two groups, each group with 16 rats. The control diet had casein as protein source (CD), and the experimental one had Ac protein concentrate (PCAcD). The diets contained 1% cholesterol. Parameters of oxidative stress in liver with CD and PCAcD were also evaluated. No significant differences were observed in serum total cholesterol, whereas LDL decreased and HDL increased (P < 0.001), and the amount of triglycerides decreased in PCAcD as compared to CD. In liver, a decrease of total cholesterol and triglycerides (P < 0.001) was observed in the experimental group in relation to control. Fatty acid synthase (FAS) activity decreased significantly in the experimental group. The mRNA of HMG-CoA reductase did not change, and mRNA of FAS decreased in rat liver fed with PCAcD compared with CD. The excretion of total lipids in feces increased with PCAcD compared to CD (P < 0.001). The activity of reactive substances to thiobarbituric acid in liver showed no significant differences between the control and experimental diets. However, total glutathione and reduced glutathione increased in PCAcD compared to CD (P < 0.001). It can be concluded that PCAcD has a hypotriglyceridemic effect, affects the metabolism of liver lipids, and increases parameters of antioxidant protection in male Wistar rats. PMID- 16380645 TI - Soy protein isolate induces CYP3A1 and CYP3A2 in prepubertal rats. AB - Feeding soy diets has been shown to induce cytochrome P450s in gene family CYP3A in Sprague-Dawley rat liver. We compared expression of CYP3A enzymes on postnatal Day 33 (PND33) rats fed casein or soy protein isolate (SPI+)-based AIN-93G diets continuously from gestational Day 4 through PND33 or the diets were switched on PND15 (n = 3-6 litters) to examine the potential imprinting effects of soy on drug metabolism. In addition rats were fed casein, SPI+, SPI+ stripped of phytochemicals (SPI-), or casein diets supplemented with the soy-associated isoflavones genistein or daidzein from weaning through PND33 to examine the hypothesis that the isoflavones are responsible for CYP3A induction by soy feeding. Feeding SPI either continuously or from weaning induced hepatic CYP3A1 and CYP3A2 mRNA, apoprotein, and CYP3A-dependent testosterone 6beta-hydroxylase activity in liver microsomes 2- to 5-fold (P < 0.05). CYP3A mRNA expression was also elevated 2- to 3-fold in the jejunum of SPI-fed rats (P < 0.05). CYP3A was not induced in livers of rats switched to casein from soy at weaning. Induction of CYP3A1 also did not occur in rats fed SPI-, but CYP3A2 mRNA and apoprotein were induced (P < 0.05) in females fed SPI-. Offspring weaned onto genistein supplemented diets had no elevation of CYP3A mRNAs or apoproteins. Weaning onto daidzein diets increased CYP3A2 mRNA and apoprotein expression in male rats (P < 0.05). These data suggest that early soy consumption may increase the metabolism of a wide variety of CYP3A substrates, but that soy does not imprint the expression of CYP3A enzymes. Effects on CYP3A1 expression appear to be primarily due to phytochemical components of SPI other than isoflavones. In contrast, consumption of soy protein and daidzein appear to be associated with the induction of CYP3A2. PMID- 16380646 TI - Accumulation of symmetric dimethylarginine in hepatorenal syndrome. AB - In patients with cirrhosis, nitric oxide (NO), asymmetric dimethylarginine (ADMA), and possibly symmetric dimethylarginine (SDMA) have been linked to the severity of the disease. We investigated whether plasma levels of dimethylarginines and NO are elevated in patients with hepatorenal syndrome (HRS), compared with patients with cirrhosis without renal failure (no-HRS). Plasma levels of NO, ADMA, SDMA, and l-arginine were measured in 11 patients with HRS, seven patients with no-HRS, and six healthy volunteers. SDMA concentration in HRS was higher than in no-HRS and healthy subjects (1.47 +/- 0.25 vs. 0.38 +/- 0.06 and 0.29 +/- 0.04 microM, respectively; P < 0.05). ADMA and NOx concentrations were higher in HRS and no-HRS patients than in healthy subjects (ADMA, 1.20 +/- 0.26, 1.11 +/- 0.1, and 0.53 +/- 0.06 microM, respectively; P < 0.05; NOx, 94 +/- 9.1, 95.5 +/- 9.54, and 37.67 +/- 4.62 microM, respectively; P < 0.05). In patients with HRS there was a positive correlation between serum creatinine and plasma SDMA (r2 =0.765, P < 0.001) but not between serum creatinine and ADMA or NOx. The results suggest that renal dysfunction is a main determinant of elevated SDMA concentration in HRS. Accumulation of ADMA as a result of impaired hepatic removal may be the causative factor initiating renal vasoconstriction and SDMA retention in the kidney. PMID- 16380647 TI - Effect of long-term somatotropin treatment on body composition and life span in aging obese Zucker rats. AB - The objective of this work was to test the hypothesis that a somatotropin (STH) induced reduction in body fat would prolong the life span of the obese Zucker rat. Two experiments were conducted. In the first experiment, male and female, lean and obese Zucker rats were treated with STH (0 or 2 mg/d bovine STH) for 4 weeks, beginning at 7 months of age. Across phenotypes, STH treatment increased the growth rate by 159%, muscle weights by 14%, and circulating insulin-like growth factor (IGF)-1 by 23%, and decreased carcass fat by 21% (P < 0.05). The second experiment was a longevity trial to determine whether these changes in body composition would increase the life span of the obese rat. Beginning at 7 months of age, individually housed, male and female, lean and obese rats were assigned to daily STH treatments (0 or 2 mg/d). Rats were monitored daily, and sick or moribund rats were euthanized and necropsied to determine existing pathologies. The average life span of the lean rats was 661 days and was unaffected by STH treatment (639 days, NS) or gender. Average life span of the vehicle-injected obese rats (435 days) was less than that of the lean group (P < 0.001). STH treatment of the obese rats resulted in a further reduction of life span (349 days, P < 0.02). The predominant pathology observed across the treatment groups was renal disease, characterized by progressive glomerulonephropathy. Thus, although exogenous STH was able to reduce carcass lipid and to increase lean tissue mass in obese rats, there was no improvement in longevity. In contrast to the hypothesis, STH actually reduced the life span of the obese rat. It is likely that STH treatment accelerated the development of progressive glomerulonephropathy in the obese rat. PMID- 16380648 TI - ECRG1, a novel candidate of tumor suppressor gene in the esophageal carcinoma, triggers a senescent program in NIH3T3 cells. AB - Esophageal cancer-related gene 1 (ECRG1) is a novel tumor-suppressor gene candidate identified from the human esophagus. Previous studies showed that ECRG1 overexpression could inhibit cell growth and induce G1 cell cycle arrest and p15(INK4b) expression by interacting with Miz-1 (Myc-interacting zinc finger protein). Such evidence suggests the alterations in ECRG1 may play an important role in tumorigenesis. To further study the biological function of the ECRG1 gene, we transfected ECRG1 into NIH3T3 cells. Expression of ECRG1 in these cells caused senescence-like changes characterized in terms of altered cell morphology, cell cycle arrest at the G1/S phase, and significantly impaired cell proliferation (P < 0.01). Moreover, NIH3T3 cells transfected with ECRG1 stained positive for SA-beta-gal staining (pH 6.0), which is a specific marker of cellular senescence. We also studied changes in telomerase activity and the related senescence genes, such as p21 and p16. The results indicated that when ECRG1 induced a senescence-like state, telomerase activity was markedly decreased (P < 0.05), and expression of p21 was distinctly increased, whereas no changes were detected in p16 and telomerase-component RNA levels. These findings suggest that ECRG1 may be of importance in murine cell senescence, promoting senescence by regulating expression of p21. PMID- 16380649 TI - T cell deficiency leads to liver carcinogenesis in azoxymethane-treated rats. AB - There is an increasing amount of evidence suggesting that T cell deficiency contributes to tumor development. However, it is unclear whether T cell deficiency leads to liver and colon carcinogenesis. The aim of this study was to investigate the role of T cells on liver and colon carcinogenesis. Athymic F344/N Jcl-rnu/- (nu/nu) rats and euthymic F344/N Jcl-rnu/+(nu/+) rats were administered the carcinogen azoxymethane (AOM) at a dose of 15 mg/kg body wt once a week for 2 weeks. At 48 weeks after the second carcinogen treatment, the rats were sacrificed, and livers and colons were examined. Apoptosis and cell proliferation were evaluated by DNA fragmentation and proliferating cell nuclear antigen assays, respectively. Wild-type p53 and members of the Jun and Fos oncogene families were detected by Western blotting. AOM treatment induced 100% liver tumor and 63.6% colon tumor incidence in T cell-deficient nu/nu rats, compared with 0% and 38.5% incidence in nu/+ rats. T cell deficiency promoted the inhibitory action of AOM on apoptosis in both liver and colon at 48 weeks. In contrast, T cell deficiency increased cell proliferation after AOM treatment in both tissues. Wild-type p53 was reduced in both tissues of T cell-deficient rats. AOM treatment induced c-Jun and c-Fos expressions in the liver but increased only Fos B in the colon, whereas T cell deficiency enhanced c-Jun overexpression in the liver. These results suggest that T cell deficiency leads to liver carcinogenesis partly by a reduction in wild-type p53 and increasing c-Jun expression in AOM-treated rats. PMID- 16380650 TI - Arachidonic acid metabolites mediate the radiation-induced increase in glomerular albumin permeability. AB - Radiation-induced renal injury is characterized by proteinuria, hypertension, and progressive decline in renal function. We have previously shown that in vivo or in vitro irradiation of glomeruli with a single dose of radiation (9.5 Gy) increases glomerular albumin permeability (P(alb)) within 1 hr. The current studies tested the hypothesis that this early radiation-induced increase in P(alb) is caused by the release of arachidonic acid and by the generation of specific arachidonic acid metabolites. Glomeruli obtained from WAG/Rij/MCW rats and cultured rat glomerular epithelial and mesangial cells were studied after irradiation (9.5 Gy, single dose). Arachidonic acid release and eicosanoid synthesis by glomeruli or cultured glomerular cells were measured after irradiation, and the effect of inhibitors of phospholipase A2 (PLA2) and cyclooxygenase (COX) on the irradiation-induced increase in P(alb) was assessed. Arachidonic acid release was demonstrated within 10 mins of irradiation of isolated glomeruli and monolayer cultures of glomerular epithelial and mesangial cells. Prostaglandin F(2alpha) (PGF(2alpha)) and PGE2 release was increased after irradiation of isolated glomeruli. Blocking arachidonic acid release or COX activity before irradiation completely prevented the increase in P(alb). COX inhibition immediately after irradiation also diminished the radiation-induced increase in P(alb). We conclude that arachidonic acid and its COX metabolites play an essential role in the early cellular changes that lead to the radiation induced increase in P(alb). Understanding of the early epigenetic effects of irradiation may lead to new intervention strategies against radiation-induced injury of normal tissues. PMID- 16380651 TI - Synergistic effect of hydrogen peroxide and elastase on elastic fiber injury in vitro. AB - This laboratory has previously shown that hyperoxia enhances airspace enlargement in a hamster model of elastase-induced emphysema. To further understand the mechanism responsible for this finding, the effect of oxidants on elastase activity was studied in vitro, using a radiolabeled elastic fiber matrix derived from rat pleural mesothelial cells. Matrix samples were treated with either 0.1%, 1%, 3%, or 10% hydrogen peroxide (H2O2) for 1 hr, then incubated with 1.0 microg/ml porcine pancreatic elastase for 2 hrs. Radioactivity released from the matrix was used as a measure of elastolysis. Results indicate that sequential exposure to H2O2 and elastase markedly enhanced elastolysis compared to enzyme treatment alone. A 22% increase in elastolysis was seen with 0.1% H2O2 (325 vs. 396 cpm; P < 0.05), whereas samples pretreated with 1%, 3%, and 10% H2O2 showed increases of 53% (274 vs. 420 cpm; P < 0.05), 71% (381 vs. 653 cpm; P < 0.01), and 38% (322 vs. 443 cpm; P < 0.01), respectively. Exposure to various concentrations of H2O2 alone (0.1% to 10%) produced only minimal elastolysis (<20 cpm). However, 1% H2O2 was capable of degrading peptide-free desmosine and isodesmosine, suggesting that exposure to this oxidant may reduce the stability of the elastic fiber matrix. With regard to lung diseases such as emphysema, H2O2 and other oxidants derived from inflammatory cells or the environment could possibly act as priming agents for elastase-mediated breakdown of elastic fibers, resulting in amplification of lung injury. PMID- 16380653 TI - Modified amalgam plugger for etchant application. PMID- 16380652 TI - Functional hypersplenism in mice induced by adoptive transfer of syngeneic spleen cells. AB - Investigation of peripheral blood cell count alterations in cases with hypersplenism, and an understanding of the relationship between splenic function and hematopoietic cell production require suitable experimental animal models. Previously described methods are either traumatic or require surgical intervention. We suggest a relatively simple method for achievement of a state mimicking hypersplenism in mice by intraperitoneal inoculation of syngeneic spleen cells. Mice were inoculated intraperitoneally with 3 x 10(7) splenocytes suspended in 0.3 ml phosphate buffered saline (PBS). After 2 months, the inoculated animals showed a progressive decrease in the peripheral white blood cell (WBC) counts and hyperplastic bone marrow that persisted until the experimental end point (7 months). Five days after inoculation of splenocytes stained with carboxy-fluorescein diacetate succinimidyl ester (CFSE), the majority of the stained cells was present in the peritoneal cavity (33%) and in the liver (13%), whereas the percentage of stained cells in the peripheral blood and the spleen cell suspension was negligible. The mitogen response of the peripheral blood mononuclear cells (PBMC) from treated mice to concanavalin A (Con A) remained unaltered. Splenocyte-inoculated mice that were further splenectomized did not show leukocytosis after splenectomy, as was observed in animals in which the spleen was removed without any pretreatment. The lack of any signs of discomfort in animals from the study group, in comparison with the visibly ill appearance and even death of mice in which hypersplenism was achieved by repeated injections of methylcellulose (MC), which served as controls, favors the convenience of the method. PMID- 16380654 TI - Modified transpalatal bar for maxillary molar intrusion. PMID- 16380656 TI - 2005 JCO Orthodontic Practice Study. Part 1: trends. PMID- 16380657 TI - Computer-aided bracket placement for indirect bonding. PMID- 16380658 TI - Molar control using indirect miniscrew anchorage. PMID- 16380659 TI - [Epidemiology of hepatocellular carcinoma in Korea]. PMID- 16380660 TI - [HBV mutations during antiviral therapy in patients with chronic hepatitis B]. PMID- 16380661 TI - [The incidence rate of hepatitis B virus surface gene variants in Korean children with immunoprophylaxis failure of perinatal infection]. AB - BACKGROUND/AIMS: Perinatal infection with hepatitis B virus (HBV) may occur despite immunoprophylaxis. One of the important mechanisms for perinatal prophylaxis failure, might include HBV surface gene variants. Therefore, we screened Korean children, in whom perinatal prophylaxis failed, for HBV surface gene variants. METHODS: Thirty-one children with perinatal HBV prophylaxis failure were selected. To amplify the major hydrophilic region of the HBV surface gene, nested PCR with primers targeted to nucleotides 237 to 706 was performed, and then sequencing was done. RESULTS: All cases were shown to be PCR positive for HBV-DNA and genotype C. Nine out of 31 (29%) with perinatal prophylaxis failure had a nucleotide substitution at the major hydrophilic region of the gene; but only two cases (6.5%) had an amino acid substitution. One case was infected by wild type and variants of I126S, and the other by wild type and S114A+I126S, respectively. CONCLUSIONS: In Korea, compared to the previous studies of other nations, gene surface variants such as G145R do not appear to play an important role in perinatal immunoprophylaxis failure. PMID- 16380662 TI - [The role of neuropsychological testing and electroencephalogram for early detection of minimal hepatic encephalopathy]. AB - BACKGROUND/AIMS: Minimal hepatic encephalopathy in patients with clinically asymptomatic chronic progressive liver disease may have adverse effects on daily activity. We evaluated the differences in the cognitive function of patients with chronic hepatitis and liver cirrhosis group according to the Child-Pugh classification. METHODS: We enrolled 61 consecutive chronic liver disease patients. We used the following study instruments: visual continuous performance test, a spatial memory test, the Wisconsin card-sorting test chosen from Neuroscan and STIM system (Study of the Usefulness of Computerized Neuropsychological Test, Neurosoft company, New York, NY, USA), a global-local processing test and an electroencephalogram (EEG). RESULTS: A significant correlation was found between neurologic abnormalities and the degree of liver disease. The result of the neuropsychological test and the EEG showed that cognitive function decreased according to the severity of chronic liver disease, especially in liver cirrhosis. Cirrhotic patients, especially in Child-Pugh C group, exhibited selective deficits in complex attention and fine motor skills as well as visual spatial perception, with preservation of memory. CONCLUSIONS: The STIM and EEG are simple, subjective and reproducible methods and may be used as early detection methods of minimal hepatic encephalopathy. PMID- 16380663 TI - [Comparison of liver abscess between diabetic patients and non-diabetic patients]. AB - BACKGROUND/AIMS: Klebsiella pneumoniae is emerging as the leading cause for liver abscess although the most common pathogen was Escherichia coli in the past. Patients with diabetes mellitus are more likely to have a pyogenic liver abscess with gas forming infection; a gas forming pyogenic liver abscess carries a higher morbidity and mortality than the non-gas forming group. This study was conducted to clarify the clinical presentation and prognostic factors for pyogenic liver abscess in diabetic patients compared with non-diabetic patients. METHODS: Medical records of 140 cases of patients treated for pyogenic liver abscess from January 1995 through January 2004 were reviewed retrospectively in detail. RESULTS: Among 140 cases of pyogenic liver abscess, underlying diabetes was present in 26.4% (37/140). The clinical presentation between the two groups was not significantly different. The most common organism for the pyogenic abscess was K. pneumoniae in both groups. A gas forming liver abscess was discovered in only the diabetic liver abscess group, 6 of 37 patients (16%). CONCLUSIONS: K. pneumoniae was the most common organism cultured in both diabetic and non diabetic liver abscess. Gas forming liver abscess was more common in diabetic patients than non-diabetic patients. Diabetic patients had more complications than non-diabetic patients. PMID- 16380664 TI - [Clinical outcome of pulmonary resections in patients with pulmonary metastasis of hepatocellular carcinoma]. AB - BACKGROUND/AIMS: Although the lung is the most common site of extrahepatic spread from hepatocellular carcinoma (HCC), the role of surgery for pulmonary metastasis remains unclear. The aim of this study was to evaluate the efficacy of pulmonary resection in patients with pulmonary metastasis from HCC. METHODS: Between July 2000 and July 2004, a total of 6 patients with pulmonary metastasis from HCC underwent curative pulmonary resections. The patients were divided into two groups (Surgery group and Non-surgery group) according to the primary treatment modality of HCC. Medical records, imaging studies, and pathologic reports of the surgical specimens were reviewed. RESULTS: Three patients in the surgery group underwent pulmonary resections for a solitary metastasis after hepatectomy for HCC, and they are all still alive. One of the 3 patients developed a tumor recurrence in the chest wall after pulmonary resection. The survival time after diagnosis of HCC were 79, 122, and 54 months, respectively. The survival time after pulmonary metastatectomy were 49, 39, and 20 months in the three patients. Another 3 patients in the non-surgery group, received a pulmonary metastatectomy; they had either a complete response HCC or partial radiologic response. These 3 patients developed recurrent disease in the liver. One of 3 patients died. The survival time after diagnosis of HCC were 153, 83, 12 months. The survival time after pulmonary metastatectomy were 51, 4, 2 months. CONCLUSIONS: The surgical resections of a solitary pulmonary metastasis from HCC in highly selected patients might be an effective treatment modalities for prolonged survival. PMID- 16380665 TI - [The efficacy of hepatic arterial infusion therapy for advanced hepatocellular carcinoma according to extrahepatic collateral feeding vessels]. AB - BACKGROUND/AIMS: Despite the poor response rate of 20-30%, hepatic arterial infusion therapy (HAIT) has been often tried for advanced hepatocellular carcinoma with portal vein tumor thrombosis or ineffective response to other treatments. The factors that predict treatment response to HAIT remain unclear. This study ascertained the response rate to HAIT based on the existence of extrahepatic collateral feeding vessels or anatomical variants. METHODS: Forty one patients received repeated HAIT using an implanted drug delivery system. Of the 41 patients, 18 patients were treated with 5-FU, epirubicin and mytomycin-C; 17 patients were treated with 5-FU and cisplatin; and 6 patients were treated with 5-FU, cisplatin and leucovorin. The patients were divided into two groups according to the existence of extrahepatic collateral feeding vessels or anatomical variants. RESULTS: Of the 41 patients, 10 patients (24.4%) showed a complete response (CR) or partial response (PR). Of 41 patients, 22 patients (group A) did not have extrahepatic collateral feeding vessel or an anatomical variant, but 19 patients (group B) did. In group A, 10 patients (45.5%) had a treatment response (CR+PR). However, only one patient (5.3%) had a treatment response (CR+PR) in group B. The response rate in group A was significantly higher than that in group B (45.5 vs. 5.3%; P=0.005). The median survival of group A was significantly longer than that of group B (10.8 vs 3.4 months, P=0.031). CONCLUSIONS: Hepatic arterial infusion therapy may be useful therapeutic option for patients with advanced HCC, especially in those that do not have extrahepatic collateral feeding vessel or anatomical variant. PMID- 16380666 TI - [The factors related to the prognosis of solitary hepatocellular carcinoma after radiofrequency ablation]. AB - BACKGROUND/AIMS: Several risk factors, such as size and location, are related to local recurrence after radiofrequency ablation (RFA) in the treatment of hepatocellular carcinoma (HCC). The objectives of this study were to clarify factors related to prognosis. METHODS: From October 1999 to December 2002, we performed RFA for 107 consecutive patients with solitary HCC. We evaluated spiral computed tomography and serum alpha-fetoprotein level every 3 months after RFA. Seven possible factors for prognosis were analyzed using the Cox proportional hazards regression model: tumor size, tumor location, age, sex, etiology, platelet count, and Child-Pugh classification. Overall survival and disease free survival rate were estimated using the Kaplan-Meier method, and differences between two groups were compared using the log rank test. RESULTS: The Kaplan Meier estimates of overall survival after radiofrequency ablation were 90.5% at 12 months, 67.4% at 24 months and 46.4% at 36 months and disease free survival were 71.4%, 46.7% and 20.9%, respectively. Using the Cox proportional hazards regression model, it was shown that with regard to overall survival and disease free survival, Child-Pugh classification (P=0.001, P=0.026) and platelet count (P<0.001, P=0.002) were statistically significant factors. The other factors did not have a statistically significant relationship to overall survival and disease free survival. CONCLUSIONS: The size and location known as local recurrence factors were not statistically significant with regard to survival and disease free survival. The Child-Pugh classification and platelet count, that reflect the liver function at the time of RFA, were significant factors for prognosis. PMID- 16380667 TI - A case of the hepatocellular carcinoma during the pregnancy and metastasis to the left atrium. AB - A 28-year-old female with a pregnant state of 29th gestational week was admitted because of nausea and vomiting. MRI showed a huge hepatocellular carcinoma. At the 36th gestational week, a normal delivery was done. Surgery of hepatocellular carcinoma was done at 20 days after delivery. After 22 months from the operation, pulmonary metastasis was found and resection of lung mass was done. About 3 years after resection, a low attenuating nodule was detected in the left atrial chamber of heart. We report a case of hepatocellular carcinoma with metastatic lesions to lung and heart in a pregnant woman. PMID- 16380668 TI - [A case of primary hepatic leiomyosarcoma with intrahepatic and abdominal subcutaneous metastasis in Behcet's disease]. AB - Primary hepatic leiomyosarcoma is a very rare tumor of the liver and primary hepatic leiomyosarcoma with Behcet's disease has not been reported previously. Behcet's disease is a multisystem disorder presenting with recurrent oral and genital ulcerations as well as ocular lesions; it has rarely been reported in association with malignant disease. We report a case of primary hepatic leiomyosarcoma with intrahepatic and abdominal subcutaneous metastasis in a patient with Behcet's disease; this is the first report of these findings in Korea. PMID- 16380670 TI - Effect of rosiglitazone on early-morning plasma cortisol levels. AB - OBJECTIVES: PPAR-gamma agonists are able to inhibit pituitary tumour development and tumoral hormonal secretion in rodents both in vitro and in vivo. Their use for treatment of Cushing Disease (CD) has been suggested but the clinical experience with the two PPAR-gamma agonists commercially available (rosiglitazone and pioglitazone) was not impressive. Short-time treatment has been proposed to be the cause of unsuccessful results on CD in humans. We report here the effect on early-morning plasma cortisol levels of a long-time treatment with rosiglitazone at the highest approved dose. METHODS: Because PPAR-gamma receptors are located in normal corticotroph cells we tested in a placebo-controlled study the influence of rosiglitazone on cortisol secretion. The study enrolled 30 newly diagnosed type 2 patients which were assigned to receive either rosiglitazone (8 mg/day) or placebo. Plasma morning cortisol (8.00 a.m.) was measured at the baseline and at the end of the study. RESULTS: Rosiglitazone vs placebo did not modify the early morning plasma levels of cortisol (13 microg/dl [3-21] vs 11 microg/dl [7-23] [median and range]) after 26 weeks of treatment. CONCLUSION: The discrepancy between in vitro and animal data on one side and clinical data on the other side warrant further investigations into the mechanisms of action of PPAR gamma agonists on ACTH secretion before other clinical studies will be conducted. PMID- 16380671 TI - Unusual presentation of hypophysitis preceding an empty sella in a 75-year-old woman. AB - A 75-year-old woman complained about progressing fatigue. She appeared somnolent, but fully oriented and in no acute distress. Her face was pale and puffy. She did not show any signs of focal neurological disease, and the remainder of the physical examination was unrevealing. Routine laboratory tests were unremarkable except for hyponatremia and mildly decreased levels of free T3 and free T4, with TSH in the normal range. Pituitary function tests demonstrated secondary adrenal insufficiency and hypothyroidism. Magnetic resonance imaging (MRI) unmasked hypophysitis with the characteristic findings of homogeneous gadolinium uptake of the pituitary and a prominent pituitary stalk ('dural tail sign', arrows in Fig. 1 A and B, sagittal and coronal views). Substitution of hydrocortisone and levothyroxine resulted in rapid and sustained improvement of all symptoms and normalisation of laboratory findings. MRI abnormalities normalized within the following six months. At follow-up three years later, MRI signs had further regressed and demonstrated an empty sella (Fig. 2 A and B). PMID- 16380672 TI - Reproductive disturbances in type 1 diabetic women. AB - Among young type 1 diabetic women disturbances of reproductive system and other related disorders are often present. The present paper, which reviews the literature of the part several years aims to present some of those disorders. Special attention is focused on menstrual irregularities, fertility and sexual problems. Type 1 diabetic women usually have a delayed menarche and an early onset of menopause than nondiabetic women. They are also at higher risk of having menstrual disturbances, such as amenorrhea and oligomenorrhea. It has been suggested that the GnRH pulse-generator in the hypothalamus is responsible for diabetic menstrual dysfunction. The risk of sexual and gestational problems is higher in type 1 diabetes than in the general population, but fertility in diabetic women seems to be similar to nondiabetics. PMID- 16380673 TI - Beta-endorphin prevents collagen induced arthritis by neuroimmuno-regulation pathway. AB - OBJECTIVE: To observe the effects and mechanisms of beta-endorphin (beta-END) preventing collagen induced arthritis (CIA) by neuroimmuno-regulating pathway. METHODS: Female wistar (Ws) rats were used in this study. CIA was induced by Native bovine type II collagen emulsified with complete Freund's adjuvant (CFA). Beta-END was administered i.p. to CIA rats every other day from the 14th day (secondary immunization) to the 35th day after primary immunization. Clinical assessments were performed by two independent, blinded examiners every other day. Pathological and radiological observations were taken on the 35th day after the primary immunization. Tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), regulated upon activation, normal T-cell expressed and secreted (RANTES), inducible NO syntheses (iNOS), matrix metalloproteinase-2 (MMP-2) and MMP-9 mRNA expression of synovium tissues of CIA rats was estimated by quantitative RT-PCR. The frequency of spleen Th1 and Th2 cells were assessed by fluorescence activated cell sorter (FACS) assay. RESULTS: Clinical manifestation of rats with CIA were significantly abrogated or ameliorated by treatment with beta-END. Beta-END treatment in vivo could down regulate mRNA expression of several pro-inflammatory cytokines, chemokines and MMPs in CIA synovial, and polarize Th1/Th2 balance to Th2. CONCLUSION: Beta-END alleviates CIA through both depressing Th1 responses and down-regulating proinflammatory and other rheumatic factors, suggesting beta-END is a promising anti-inflammatory and anti-rheumatic agent in treating CIA. PMID- 16380674 TI - Polymorphism of the glycoprotein Ia and IIIa in the group of women in childbirth does not correlate with an increased risk of developing thrombosis. AB - OBJECTIVE: The aim of the study was to evaluate the prevalence of the GP Ia and GP IIIa polymorphisms in the group of women in labor, and to assess the risk of thrombosis associated with their occurrence. DESIGN AND SETTINGS: 245 women in labor hospitalized between 1.01.2001 and 31.12.2003 r. were enrolled in the study. Patients were qualified for the study if detailed physical exam and past medical history excluded existence of known risk factors predisposing to thrombosis. Study group was composed of 72 women in childbirth, which at some point during current pregnancy or in early labor were diagnosed with thrombosis, and control group included 173 women in labor randomly picked from the group of patients with uncomplicated pregnancies. Polymorphic regions of platelets glycoprotein were detected using genotyping methods based on polymerase chain reactions (PCR). RESULTS: 1.72% of patients were found to have thrombosis. The thrombosis was located in the venous system in 97.2% of cases. Arteries were affected in two patients (2.7%). Prevalence of individual platelets glycoprotein mutations did not differ between controls and study group. In both groups platelets glycoprotein polymorphisms moderately pro-thrombotic A1/A2 and C/T dominated, and the least numerous were strongly pro-thrombotic A2/A2 and T/T. CONCLUSIONS: Our results did not show the causative relationship between the existence of platelets GP IIIa/GP Ia mutations and venous system thrombosis in the women in labor. Probably presence of pro-thrombotic mutations of platelets glycoprotein in this group of patients does not represent the increased risk of thrombosis. PMID- 16380675 TI - Effects of different antioxidants on lipid peroxidation in brain homogenates, induced by L-thyroxine administration in rats. AB - OBJECTIVE: It is known that thyrotoxicosis induces lipid peroxidation (LPO). In contrast, propylthiouracil (PTU), a thyrostatic drug is a well-known antioxidant. Also melatonin has been shown to protect against free radical-induced neuronal destruction. At the same time, it is generally accepted that the brain is the most vulnerable tissue to oxidative stress. METHODS: The goal of the study was to examine the components of LPO, i.e., conjugated dienes (CD), malondialdehyde (MDA) and Schiff's bases (SB), in the brain of male Wistar rats. Two experiments were performed, with two control groups created for each experiment: Group 1- intact animals and Group 2--animals injected with 0.9% NaCl. In Experiment I, the animals received L-thyroxine (L-T4) in a dose of 100 microg/kg BW, i.p., daily, for two weeks (Groups 3-5). After one week of L-T4 treatment, the following agents were added during a subsequent week: Group 4--PTU in drinking water (45 mg/kg BW/day); Group 5--melatonin (5 mg/kg BW, daily). In Experiment II, lasting 7 days, the animals were divided into the following groups: Group 1--intact animals; Group 2--animals injected with 0.9% NaCl; Group 3--PTU in drinking water (45 mg/kg BW/day); Group 4--melatonin (5 mg/kg BW, daily). RESULTS: In Experiment I, we observed a significantly higher SB level in saline treated animals and a significant increase in both CD and SB levels in rats treated with L-T4, compared to those in non treated control. CD levels were also elevated in rats treated with L-T4, compared to values in the saline only treated animals. Melatonin and PTU reduced CD levels and melatonin diminished SB levels, as compared to those in L-T4-treated rats. In Experiment II, we observed significantly higher CD, SB and MDA levels in saline treated rats, when compared to respective values in non treated control. Melatonin decreased CD levels, when compared to CD levels in both the non treated and saline injected controls. Additionally, melatonin reduced SB levels relative to change in the brains of saline treated rats. Furthermore, PTU decreased CD levels in brain homogenates compared to non-treated animals. CONCLUSIONS: (1) L-thyroxine administration stimulates LPO in the rat brain; (2) All the examined antioxidants decrease LPO in L-T4-administered animals; (3) All the examined antioxidants reduce the basal LPO; 4) Stress, when induced by handling, intensifies oxidative processes in the organism. PMID- 16380676 TI - The occurrence of genital types of human papillomavirus in normal pregnancy and in pregnant women with pregestational insulin dependent diabetes mellitus. AB - OBJECTIVE: The aim of the study was to evaluate the prevalence of human papillomavirus, both types of a low oncogenic risk (HPV 6, 11) and a type carrying a high oncogenic risk (HPV 16) in the genital tract of the pregnant patients, their venous blood, the cord blood and the oral cavities of the neonates. Normal pregnant women and pregnant women with insulin dependent diabetes mellitus (IDDM) diagnosed before pregnancy were included in the study. DESIGN: The study group consisted of 15 pregnant women aged 22 to 32 years with IDDM diagnosed before pregnancy. The control group consisted of 30 patients aged 18 to 38 years, with normal pregnancy. The DNA of HPV types 6, 11 and 16 was studied in the discharge from the cervical canal, the maternal venous blood, the cord blood and the buccal smear obtained from the neonates. To detect of viral DNA the PCR was used. SETTING: A university teaching hospital delivering approximately 2000 women annually. RESULTS: Human papillomavirus (HPV) was found in 12 (26.7%) of the 45 pregnant women. Of the 15 patients with pregestational IDDM the DNA of HPV was detected in four (26.7%) of the patients. The DNA of HPV types 6 and 11 was found in three (20%) patients. The DNA of HPV type 16 was detected in one pregnant patient in the study group (6.67%). Of the 30 control patients, HPV DNA was detected in eight (26%). In two (6.6%), infection with oncogenic and non-oncogenic types of HPV was diagnosed. The DNA of HPV types 6 and 11 was found in six (20%) subjects. Of the 30 control patients, the DNA of HPV type 16 was detected in four (13.3%). The transmission of HPV from HPV positive mother to fetus was found in 50% of cases. CONCLUSIONS: (i) There was a similar level of occurrence of HPV infections in pregnant women with IDDM when compare with normal pregnancy. (ii) High percentage of HPV transmission from mother to neonate was determined. (iii) The cesarean section probably does not protect the neonate from HPV infection. (iv) There is a suggestion that fetus may be affected by HPV infection during intrauterine life. PMID- 16380677 TI - Clinical significance of the insulin-like growth factor I gene promoter (P1) polymorphism in thyroid nodular disease. AB - OBJECTIVES: Due to the recent increase of incidence of thyroid nodules and the known risk of malignant transformation, there is an elevated risk of thyroid cancer in Poland. Several approaches, including molecular, have been proposed to support fine needle aspiration biopsy in the early detection of malignant lesions. Although the IGF-I system in thyroid cancer has been studied, little is known about the gene and its promoter structure changes. Our aim was to assess, whether the analysis of the IGF-I gene promoter region and 5'UTR exon 1 structure may be useful in assessing the risk of thyroid carcinoma. MATERIAL: Our study included 46 patients that underwent strumectomy due to a presence of thyroid nodules. METHODS: All patients underwent clinical examination and laboratory investigations to assess their thyroid structure and function. Tissues obtained during the surgery were used for DNA extraction, PCR, SSCP and direct sequencing. RESULTS: Among 46 patients, 14 had a nucleotide difference in one of the examined regions. In our study we revealed no significant difference between carcinomatous and non-carcinomatous groups of patients in terms of presence of nucleotide change, but Fisher's exact test gave a significant result in terms of the efficacy of detecting follicular adenoma. Moreover, the patients with nucleotide change had thyroid glands significantly smaller in volume. CONCLUSIONS: We conclude that the molecular analysis of the IGF-I gene promoter is thought to be of a functional significance, but probably could not be considered useful in the assessment of risk of thyroid cancer in thyroid nodules. PMID- 16380678 TI - Academic mothers have a pronounced seasonal variation in their offspring sex ratio. AB - OBJECTIVES: Environmental and socio-demographic factors can influence the variation of the human sex ratio at birth (SRB = the ratio of males to males plus females). In particular, findings of seasonal, parental education, birth order, and maternal age effects on the SRB are not always in agreement, and a number of works report minimal variation. SETTING: Here, we investigated the seasonality of SRB in academic and non-academic mothers employed at the University of Vienna, and who gave birth between 1963 and 2000 (n = 1932 births). METHODS: All data were available from an anonymous employee database. RESULTS: Both groups, academic and non-academic mothers do not differ between their overall SRB. In academic mothers the SRB is significantly (P = 0.004) increased during the springtime and decreased during the summertime. Although in non-academic mothers the trend is comparable, it is far less pronounced and not significant (P = 0.345). When a multiple logistic model was applied to the data of academic mothers the only significant influencing factor on the SRB is the season, while birth order of children and mothers' age at childbirth has no effect. None of the three independent variables influence the SRB in non-academic mothers. MAIN FINDINGS: These findings suggest a more flexible SRB rate in academic mothers than in non-academics within the seasons. CONCLUSION: We conclude that the significant seasonal variation of the SRB in academic women cannot be merely interpreted as an effect of socio-economic status but more likely as the interaction between socio-economic and environmental working conditions. PMID- 16380679 TI - Plasma beta carotene in Alzheimer's disease. Association with cerebrospinal fluid beta-amyloid 1-40, (Abeta40), beta-amyloid 1-42 (Abeta42) and total Tau. AB - We studied the plasma beta carotene concentrations in 40 Alzheimer's disease patients and the association with cerebrospinal fluid beta-amyloid 1-40, (Abeta40), cerebrospinal fluid beta-amyloid 1-42 (Abeta42) and cerebrospinal fluid total Tau. We found that patients with plasma beta carotene levels below the 25th percentile had 55% reduced ratios of Abeta40/Tau and 51% reduced ratios of Abeta 40/Abeta 42 compared with patients in the highest quartile. Mean Tau concentrations in the lowest quartile of plasma beta-carotene levels were 74% higher compared with the highest quartile of plasma beta-carotene levels. Thus, we could demonstrate an statistically significant association between beta carotene levels in plasma and neurochemical markers in the cerebrospinal fluid of Alzheimer's disease patients. PMID- 16380680 TI - Cyclin D1 expression in primary thyroid carcinomas. AB - OBJECTIVES: The aim of the study was to demonstrate and evaluate the expression of cyclin D1, a protein connected with a cell cycle, by means of the immunohistochemical method in malignant thyroid neoplasms. The purpose of the analysis of the results was to explain the relation between cyclin D1 in thyroid cells and neoplasm transformation. MATERIAL AND METHODS: The study was conducted on thyroid neoplasms from 35 patients who were diagnosed with the thyroid carcinoma (30 women and 5 men). Detection DAKO LSAB + system was applied with use of monoclonal antibodies against cyclin D1. The results of immunohistochemical reaction was described as an index (percentage of cells showing a characteristic brown color in 1000 counted cells). As a positive result of reaction an intensive brown color of carcinomas cellular nuclei was acknowledged. RESULTS: The mean value of cyclin D1 expression index in papillary carcinoma was 14.44% +/- 9.37, in medullary carcinoma 27.35% +/- 5.40, in nonpapillary carcinomas originating from A cells 18.0% +/- 10.20. The results were statistically analyzed. In medullary carcinoma the highest values of positive cells cyclin D1 index were revealed. CONCLUSIONS: The results obtained encourage continued studies on cyclin D1 expression in thyroid neoplasms and a more accurate analysis with a larger number of cases. Perhaps the index of this protein will become a recognized prognostic marker in thyroid neoplasms or an objective risk factor of the thyroid epithelial cells neoplastic transformation. PMID- 16380681 TI - Kerosene poisoning in childhood: a 6-year prospective study at the Princess Rahmat Teaching Hospital. AB - PURPOSE: Poisoning due to drugs is influenced to a large extent by the population's socioeconomic and cultural status. The purpose of our study was to determine and present the pattern of poisoning with kerosene in Princess Rahmat hospital, Irbid, Jordan, and to assess the effects of variables such as age, season, sex and agent on poisoning frequency in Princess Rahmat hospital, northern Jordan. METHODS: This retrospective cross-sectional study was performed on 122 cases of poisoning with kerosene who were hospitalized in Princess Rahmat hospital, northern Jordan, during a six-year period from 1996 to 2001. The data collected include: age, sex, seasonal variation, material ingested, symptomatic or not on admission, time and place of ingestion, history of similar problem among the patient or his siblings, whether vomiting was induced by the parent before seeking medical help or not and the outcome regarding mortality and morbidity. Chi-square and values were considered for the statistical analysis. RESULTS: In a 6-year prospective study of kerosene poisoning in children admitted to the Rahmat Hospital, between January 1996 and December 2001, a total of 122 children (under 11 years of age). Among the children 48 (39.3%) were girls and 74 (60.6%) were boys. Majority (80.1%) were below 2 years. The largest group of patients was admitted during the summer months. The most commonly observed symptoms were cough (67.2%), tachypnea (56.5%), fever (54.1%), and vomiting (27.8%). About one third of the patients showed signs of central nervous system (CNS) impairment, including drowsiness, restlessness, stupor, and convulsions. CONCLUSIONS: 1) There is a higher risk of kerosene poisoning during the hot months of the year; 2) the respiratory system is the main target organ affected; 3) pneumonia is in most cases interstitial and bilateral; 4) vomiting after hydrocarbon ingestion is related to the rate of development of pneumonia; 5) symptoms of CNS impairment were correlated with hypoxemia, pneumonia, and fever; and 6) CNS toxicity may occur without hypoxemia, concurrent pulmonary pathology, or other pathology. PMID- 16380682 TI - Different manifestation of depressive disorder in the elderly. AB - OBJECTIVE: To compare the clinical manifestation of depressive disorder in elderly, and younger adults. METHODS: To compare the clinical manifestation of depressive disorder, we evaluate 46 elderly (33 female, and 13 male, mean age 71.1) and 60 younger adults (40 female, and 20 male, mean age 44.5 years). All patients suffering from depressive disorders according to ICD-10. For evaluation and comparison of depressive symptomatology we used the HAM-D-17. The results analysed by the SPSS. RESULTS: The clinical manifestation of depression is different in the elderly. Elderly depressed patients compared with their younger counterparts, scored significantly less in Depressed mood, but significantly higher in Work and activities, Retardation, Somatic symptoms-general, Hypochondriasis, Insomnia-middle, Insomnia-late, Anxiety-somatic, and Somatic symptoms-gastrointestinal. On the other hand, younger patients scored significantly higher in Feelings of guilt, and Genital symptoms. CONCLUSIONS: Clinical presentation of depressive disorder is different in the elderly, depressed mood is often absent or masked. Anxiety, somatization, and hypochondriasis are more often present in the elderly depressed patients than in younger patients. The elderly people are also more likely than their younger counterparts to complain of insomnia. PMID- 16380683 TI - Influence of personality disorder on the treatment of panic disorder--comparison study. AB - Most clinicians tend to believe that the occurrence of the anxiety disorder in comorbidity with a personality disorder often leads to longer treatment, worsens the prognosis, and thus increasing treatment costs. The study is designed to compare the short-term effectiveness of combination of cognitive behavioral therapy and pharmacotherapy in patient suffering with panic disorder with and without personality disorder. METHOD: We compare the efficacy of 6th week therapeutic program and 6th week follow up in patients suffering with panic disorder and/or agoraphobia and comorbid personality disorder (29 patients) and panic disorder and/or agoraphobia without comorbid personality disorder (31 patients). Diagnosis was done according to the ICD-10 research diagnostic criteria confirmed with MINI and support with psychological methods: IPDE, MCMI III and TCI. Patients were treated with CBT and psychopharmacs. They were regularly assessed in week 0, 2, 4, 6 and 12 by an independent reviewer on the CGI (Clinical Global Improvement) for severity and change, PDSS (Panic Disorder Severity Scale), HAMA (Hamilton Anxiety Rating Scale), SDS (Sheehan Disability Scale), HDRS (Hamilton Depression Rating Scale), and in self-assessments BAI (Beck Anxiety Inventory) and BDI (Beck Depression Inventory). RESULTS: A combination of CBT and pharmacotherapy proved to be the effective treatment of patients suffering with panic disorder and/or agoraphobia with or without comorbid personality disorder. The 12th week treatment efficacy in the patients with panic disorder without personality disorder had been showed significantly better compared with the group with panic disorder comorbid with personality disorder in CGI and specific inventory for panic disorder--PDSS. Also the scores in depression inventories HDRS and BDI showed significantly higher decrease during the treatment comparing with group without personality disorder. But the treatment effect between groups did not differ in objective anxiety scale HAMA, and subjective anxiety scale BAI. PMID- 16380684 TI - Early effects of treatment on regional cerebral blood flow in first episode schizophrenia patients evaluated with 99Tc-ECD-SPECT. AB - OBJECTIVES: Altered regional cerebral blood flow (r-CBF) is mentioned as one of possible etiological factors or as a consequence of schizophrenia. Perfusion scintigraphic brain studies with SPECT (Single Photon Emission Computed Tomography) and PET (Positron Emission Tomography) can detect areas of the brain with altered perfusion during the disease. Conflicting data exist on follow-up after treatment. The aim of our study was to evaluate r-CBF in acute first episode schizophrenia and early effect of antipsychotic drugs on r-CBF. DESIGN AND SETTING: Clinical criteria for schizophrenia were met according to International Classification of Diseases-10th Edition (ICD-10). Psychic status and severity of the disease in each patient were evaluated with psychiatric interview, Positive and Negative Syndrome Scale (PANSS) and Clinical Global Impression (CGI) on the same day as the scintigraphic study. Brain perfusion SPECT was performed 20 minutes after 99mTc-ECD application. R-CBF was measured in 9 first-episode schizophrenic male patients 2-7 (median 6) days after the beginning of antipsychotic treatment and 8-15 (median 9) weeks later. Semiquantitative analysis of r-CBF was performed with cerebellum as the reference region. R-CBF was evaluated for 12 regions in each study. RESULTS: Decrease in perfusion in comparison to cerebellum was detected in left (90%) and right (93%) frontal lobes. Increase in perfusion between the first (t1) and the second (t2) investigation was significant (p<0.05) for both frontal regions (left 94%, right 97%). Significant decrease in PANSS (p<0.05) and CGI (p<0.001) scores was noted and correlated well with the increase of r-CBF. CONCLUSIONS: Our findings implicate that patients with first-episode schizophrenia have significant left hypoperfusion relative to the right in dorsolateral frontal lobes. Increased blood flow was observed bilaterally in dorsolateral frontal lobes after 10 weeks of antipsychotic medication. Significant amelioration of r-CBF correlated with clinical improvement. PMID- 16380685 TI - The alliance theory of homosexual behavior and the perception of social status and reproductive opportunities. AB - OBJECTIVES: The alliance theory holds that homosexual behavior in humans may have evolved because it reinforced same-sex alliances which contributed to survival and reproduction. The present study was designed to test this evolutionary hypothesis in view of the strongly negative perception of homosexual behavior. It was predicted that targets who engaged in homosexual behavior would be perceived as likely to achieve greater social status and reproductive opportunities when the behavior reinforced an alliance that led to increased social opportunities. METHODS: Three hundred sixteen men and women read scenarios in which a target engaged in homosexual behavior and then answered questions about the target's future social status and reproductive opportunities as measured by number of sexual partners. The data were analyzed in two 3 (social outcome: positive vs. neutral vs. negative) x 2 (gender of target) x 2 (gender of participant) analyses of variance. RESULTS: The results supported the predictions and demonstrated that targets were perceived as likely to have the greatest social status and reproductive opportunities in the positive outcome condition. CONCLUSION: The alliance theory of homosexual behavior may have heuristic value for the evolutionary study of homosexual behavior and its perception. PMID- 16380686 TI - Testosterone levels and histological features of reproductive glands in adult male rats treated neonatally with tamoxifen. AB - OBJECTIVE: In this study we examined the possibility that demasculinization produced by the neonatal administration of tamoxifen is accompanied by a decline in plasma levels of testosterone during adulthood. METHODS: Wistar male rats received either a treatment with 12.5 microg/kg of tamoxifen during the first eight days of age or a treatment with 100 microg/kg of tamoxifen for five days. Each treatment had their respective control group. During adulthood their masculine sexual behavior was analyzed. At 8 months of age, males were killed by decapitation, trunk blood was collected and peripheral glands were dissected and weighed. Testosterone levels were measured by HPLC. Histological analysis of peripheral glands was performed. RESULTS: Both neonatal tamoxifen treatments significantly decreased male sexual behavior when compared to control values. In addition, both treatments also showed a significant decrease in testicular weight when compared to control groups, as well as a decrease in seminal vesicle weight. In the microscopic analysis, a significant decrease in the diameter of the seminiferous tubules was observed, especially in the animals treated with 100 microg/kg of tamoxifen. However, no differences were observed between tamoxifen treated and control animals concerning plasma levels of testosterone. CONCLUSION: The present results indicate that behavioral manifestations and changes in peripheral reproductive organs that accompanied demasculinization are not due to a deficit in testosterone secretion. PMID- 16380687 TI - Hyperthyroidism causes lipid peroxidation in kidney and testis tissues of rats: protective role of melatonin. AB - OBJECTIVE: The present study aimed at determining how 3-week intraperitoneal melatonin administration affected oxidative stress caused by experimental hyperthyroidism. MATERIALS AND METHODS: The study was conducted on 30 male rats of Sprague-Dawley species. The experimental animals were divided to 3 groups (control, hyperthyroidism and hyperthyroidism+melatonin). The supplementation was continued for 3 weeks after which the animals were sacrificed and tissue malondyaldehyde (MDA) and glutathione (GSH) levels were determined. RESULTS: MDA levels in kidney and testis tissues in hyperthyroidism group were higher than those in control and hyperthyroidism+melatonin administered groups (p<0.001) and levels in hyperthyroidism+melatonin administered group were higher than those in the control group (p<0.001). The highest GSH levels were obtained in hyperthyroidism+melatonin-administered group (p<0.001) and GSH levels in hyperthyroidism group were higher than those in the control group (p<0.001). CONCLUSION: Results of the study demonstrate that hyperthyroidism induced by 3 week L-thyroxine administration increased oxidative stress in kidney and testis tissues and that although melatonin administration inhibited this stress to a certain extent, it could not bring the stress down to the level in controls. PMID- 16380688 TI - Pinealectomy inhibits antioxidant system in rats with hyperthyroidism. AB - OBJECTIVE: Thyroid hormones regulate energy metabolism and act on the mitochondria, which is an important source of free radicals in the cell. Reactive oxygen types play a significant role in physiological mechanisms, but in excessive amounts they can cause oxidative damage in molecules. The aim of the present study was to determine levels of lipid peroxidation caused by induced hyperthyroidism in cerebral, hepatic and cardiac tissues of pinealectomized rats. METHODS: Experimental animals used in the study were allocated to three groups as general control group, hyperthyroidism-sham pinealectomy group and hyperthyroidism-pinealectomy group. GSH and MDA levels in cerebral, hepatic and cardiac tissues were evaluated at the end of the 3-week study period. RESULTS: It was found that MDA levels in cerebral, hepatic and cardiac tissues were the highest in hyperthyroidism and pinealectomy group and that these values were higher in hyperthyroidism-sham pinealectomy group than in the control group (p<0.001). It was seen that tissue GSH levels significantly increased in hyperthyroidism-sham pinealectomy group (p<0.001) and that the increase in hyperthyroidism and pinealectomy group was higher than the increase in the control group only (p<0.001). CONCLUSION: Results of our study show that MDA and GSH levels in cerebral, hepatic and cardiac tissues increased due to hyperthyroidism and that the increase in MDA levels became more evident and GSH levels were significantly suppressed after pinealectomy. PMID- 16380689 TI - Increased cytochrome oxidase activity in adrenal glands of thioacetamide cirrhotic rats. AB - OBJECTIVES: Cytochrome oxidase activity has been determined in the adrenal glands of thioacetamide (TAA)-cirrhotic rats. MATERIAL AND METHODS: Two groups of animals (Control group; n=10 and TAA group; n=11) of three months evolution were used to study the cytochrome oxidase activity in the suprarenal cortex (glomerular, fascicular and reticular layers) and medulla. TAA was administered orally and cytochrome oxidase (COX) activity was assayed by an immunohistochemical technique. RESULTS: In TAA-cirrhotic rats, COX activity increases in the cortex fascicular layer (221 +/- 1.79 vs 181.9 +/- 2.75; p<0.001) as well as in the medulla (146.6 +/- 1.72 vs 129 +/- 3.09; p<0.001). CONCLUSION: These results make it possible to consider the existence of hypothalamic-pituitary-adrenal-axis and adrenomedullary sympathetic system hyperactivity, both peripheral limbs of the stress system, in this experimental model of cirrhosis. PMID- 16380690 TI - In chronic fatigue syndrome, the decreased levels of omega-3 poly-unsaturated fatty acids are related to lowered serum zinc and defects in T cell activation. AB - There is now evidence that major depression is accompanied by decreased levels of omega3 poly-unsaturated fatty acids (PUFA), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). There is a strong comorbidity between major depression and chronic fatigue syndrome (CFS). The present study has been carried out in order to examine PUFA levels in CFS. In twenty-two CFS patients and 12 normal controls we measured serum PUFA levels using gas chromatography and mass spectrometry. We found that CFS was accompanied by increased levels of omega6 PUFAs, i.e. linoleic acid and arachidonic acid (AA), and mono-unsaturated fatty acids (MUFAs), i.e. oleic acid. The EPA/AA and total omega3/omega6 ratios were significantly lower in CFS patients than in normal controls. The omega3/omega6 ratio was significantly and negatively correlated to the severity of illness and some items of the FibroFatigue scale, i.e. aches and pain, fatigue and failing memory. The severity of illness was significantly and positively correlated to linoleic and arachidonic acid, oleic acid, omega9 fatty acids and one of the saturated fatty acids, i.e. palmitic acid. In CFS subjects, we found significant positive correlations between the omega3/omega6 ratio and lowered serum zinc levels and the lowered mitogen-stimulated CD69 expression on CD3+, CD3+ CD4+, and CD3+ CD8+ T cells, which indicate defects in early T cell activation. The results of this study show that a decreased availability of omega3 PUFAs plays a role in the pathophysiology of CFS and is related to the immune pathophysiology of CFS. The results suggest that patients with CFS should respond favourably to treatment with--amongst other things--omega3 PUFAs, such as EPA and DHA. PMID- 16380691 TI - Substantial reaction between histamine and malondialdehyde: a new observation of carbonyl stress. AB - OBJECTIVES: To explore the mechanism of carbonyl stress-related toxification on neurotransmitter histamine and the potential de-carbonylation function of histamine. METHODS: The reaction mixture of malondialdehyde (MDA) and histamine (HA) at pH7.4, 37 degrees C was assayed by high performance liquid chromatography (HPLC), spectrophotometry and spectrofluorometry. The reaction products were assayed by LC/MS. RESULTS: In physiological condition, the reaction of MDA and HA yielded two products: a nonfluorescent enaminal derivatives and a fluorescent 1,4 dihydropyridine adducts. The fluorescence maxima of the fluorescent products (Ex. 393 nm/Em. 464 nm) were similar to those of lipofuscin pigment. The fluorescence intensity of reaction mixture was in direct proportion to the MDA concentration. CONCLUSIONS: HA can react with MDA to form stable products, a non-fluorescent enamine (product 1) and a fluorescent 1,4-dihydropyridine (product 2) which are ceroid/lipofuscin-related adducts. The reaction of HA with MDA may reveal toxic effect of unsaturated carbonyls in the brain and may reflect a novel de carbonylation function of histamine under various pathological conditions. PMID- 16380692 TI - Neonatal hypothyroidism-induced changes in rat testis size, dependence on temperature. AB - OBJECTIVE: Effects of transient neonatal hypothyroidism (HPOT) on adult testis size and serum hormone profiles were evaluated in the Charles foster strain of rats, maintained under the temperature of 21 degrees C (HPOTL) or of 34 degrees C (HPOTH). METHODS: Hypothyroidism was induced in suckling pups in preweanling period (1-21 days), by administering 0.1% 6-propyl-2-thiouracil in drinking water to mothers. Body mass and testis characteristics, the steroidogenic potential [measured by histochemical localization of 3alpha-, 3beta- and 17beta hydroxysteroid dehydrogenases (HSDHs)] and serum concentrations of thyroid stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), luteinizing hormone (LH), testosterone (T) and corticosterone (Cort) were evaluated on the 35th, 45th, 60th and 90th day of age. RESULTS: The HPOTH rats showed lower testis masses, while the HPOTL rats showed higher testis masses and lower body masses after 90 days. Histologically, the testes of the HPOTH rats demonstrated increased germ cell degeneration after 35 and 45 days and reduced tubular size, germ cell numbers and sperm density after 90 days. In contrast, the testes of the HPOTL rats showed reduced tubular diameters after 35 and 45 days, and increased tubular diameters, germ cell numbers and sperm density after 90 days. Serum TSH, T3, T4, LH and T concentrations and 3beta- and 17beta-HSDH activities were reduced in both groups of the HPOT rats after 35 and 45 days. Intratubular steroidogenesis and a prominent triangle up5 pathway were also found in the HPOT animals. CONCLUSION: The temperature has a definite influence on the thyroid hormone action, postnatal growth and function of rat testes. PMID- 16380693 TI - Cycle dependent expression of endometrial metallothionein. AB - OBJECTIVES: Molecular changes observed in endometrium with respect to menstrual cycle changes seem to be crucial for the reproductive function. Accumulation of cytotoxic cells increases the exposure of endometrial cells to apoptosis. Protection against apoptosis may be reached by endometrial cells by self molecular regulation. Metallothionein was suggested to participate in this process. The aim of our study was to evaluate endometrial MT immunoreactivity with respect to the menstrual cycle phases. MATERIALS AND METHODS: MT expression was assessed using immunohistochemistry method in 47 endometrial tissue samples derived from randomly selected patients with respect to the menstrual cycle phases--proliferative and secretory with distinguishing early, mid and late subphases in each. RESULTS: MT expression changes were observed respectively to hormonal fluctuations with the highest level during mid secretory phase and its respective decrease during the early, late secretory and mid proliferative menstrual cycle phases. The lowest MT immunoreactivity level was disclosed during early proliferative phase. CONCLUSION: Significant differences in MT expression observed in endometrium with respect to menstrual cycle changes might suggest MT participation in endometrial cells protection against apoptosis. PMID- 16380694 TI - Antiandrogenic activity of Ruta graveolens L in male Albino rats with emphasis on sexual and aggressive behavior. AB - BACKGROUND: The Ruta graveolens L. is currently used by the Jordanian population systemically for its antispasmodic, diuretic, sedative, and analgesic effects and externally for its antirheumatic effect. OBJECTIVE: To study plant effects on the reproductive system and fertility using adult male albino rats with special emphasis on the aggressive behavior and sex behavior. The aqueous extracts of Ruta graveolens L. solution was fed orally to male albino rats at a dose of 500 mg/kg body weight for 60 days. RESULTS: This dose induces a significant decrease in the weight of reproductive organs (P<0.01) when compared to controls. The sperm motility and density in cauda epidydimides and testicular ducts were significantly decreased (P<0.01). A significant decreased (P<0.001) in spermatogenesis activity is observed in somniferous tubule. Treated rats testicular cell population showed a decrease in number of spermatocytes and spermatids (P<0.001) when compared to controls. Serum hormonal assay indicated a decrease in testosterone and follicular stimulating hormone levels in treated rats. A decrease in number of female rats impregnated by males receiving treatment was observed and demonstrated by a decrease in the implantation sites and viable fetuses number (P<0.01). The ingested extract also suppresses the sexual behavior in adult male rats expressed by a prolongation of first mount time, increase in intromission latency, decrease in intromissions number, and prolongation of the post-ejaculatory interval. This led to reduction of the ejaculation time and increase of the post ejaculatory intervals. Ingestion of R. graveolens markedly abolished aggressive behavior parameters in adult male treated rats namely, a suppression in lateralization, boxing bouts and ventral presenting postures. CONCLUSION: The aqueous extracts of Ruta graveolens L might have adverse effects on territorial aggression and sexual behavior in male albino rats. PMID- 16380695 TI - Reproductive toxic effects of Topamax ingestion in female Sprague-Dawley rats. AB - PURPOSE: The objectives of this study is to investigate the toxic effects of Topamax (100 mg/kg/body weight) on the reproductive system after administration to female Sprague-Dawley rats weighing 250-300 g for two time periods 4 and 12 weeks. METHODS: Twenty adult female rats were divided into two groups and exposed to Topiramate diet at a concentration of 100 mg/kg/body weight for two periods of time. First group containing 10 rats received treatment for 4 weeks and a second group of 10 rats received the same dose of treatment for a period of 12 weeks and compared with twenty non-exposed female rats received vehicle treatment. Female rats were allowed mating with males after 10 days prior to the last administration dose. Animals were autopsied under light anesthesia after mating and several parameters were determined including: number of pregnant rats, body and reproductive organ weight, number of implantation sites, viable fetuses, and resorption sites. Assessment of pregnancies in females was measured and the significance of these results was calculated using student's 't' and Chi-square tests. RESULTS: The effect of Topamax exposure on fertility was assessed in terms of pregnant rats number, implantation sites, viable fetuses, and resorption sites. Exposure to Topamax for 4 weeks did not have much effect on fertility. Significant decrease in the relative ovarian weights and embryo weights in rats exposed to Topamax were observed. Exposure to Topamax for a 12 weeks resulted in a reduction in the percentage of pregnancies and in the number of implantation sites, when compared with controls in both treatment periods. Rats receiving 12 weeks treatment showed an increase in ovarian weights and a decrease in viable fetuses number. These results indicate that long-term exposure of female rats to Topamax causes adverse effects on the reproductive system and fertility. CONCLUSION: The results of the current study suggest that ingestion of Topamax by adult female rats causes adverse effects on fertility and reproduction. PMID- 16380696 TI - ACTH responses to somatostatin, valproic acid and dexamethasone in Nelson's syndrome. AB - OBJECTIVE: Pituitary tumours occurring in patients bilaterally adrenalectomized because of Cushing's disease (Nelson's syndrome) are frequently invasive and a complete their resection is not possible in most of them. Administration of the drugs decreasing ACTH secretion could be helpful in such unresectable tumours. We tried to evaluate the influence of somatostatin and valproic acid, compared to dexamethasone, in short-term studies, on plasma ACTH levels in Nelson's syndrome (NS). MATERIAL AND METHODS: Basal ACTH levels were determined within 18 h after last dose of hydrocortisone and next, 1 and 2 hours following oral administration of 20 mg of hydrocortisone. Somatostatin was injected s.c. in two patients with NS while sodium valproate and dexamethasone were administered orally for three days in three patients with NS (two with an invasive pituitary tumour and one with a localized, intrasellar adenoma). The blood for ACTH and cortisol determination was drawn before the tests (two hours after 20 mg of hydrocortisone ingestion) as well as 1 and 2 hours following somatostatin injection and after 3 days of valproic acid or dexamethasone administration. RESULTS: High plasma ACTH levels were found before the tests. Somatostatin lowered ACTH levels in both patients, more effectively in the patient with non-invasive pituitary adenoma. Valproic acid decreased moderately ACTH concentration in two patients, while following dexamethasone administration a fall in ACTH levels was observed in all three patients, the most evident in the patient with a non-invasive Nelson's adenoma. CONCLUSION: Somatostatin seemed to be more effective in its inhibitory action on ACTH secretion than valproic acid, thus its administration in invasive cases of NS could be tried as a supplementary method to neurosurgery. The response to dexamethasone administration indicates that a feed-back regulation, although impaired, exists in these cases. PMID- 16380697 TI - Decreased melatonin nocturnal concentrations in hemodialyzed patients. AB - OBJECTIVES: In spite of broad interest, intensive studies on function of melatonin have not yielded much information about relationships between this hormone and kidneys in health, and particularity, in disease. There are only a few studies dealing with melatonin concentrations in renal diseases, mainly performed in hemodialyzed patients with end-stage renal disease (ESRD). Moreover, the most melatonin assays were performed during the daytime, and the results are conflicting. Therefore, the aim of the present study was to determine the circadian melatonin profiles in patients ESRD before and after hemodialysis. MATERIAL AND METHODS: Thirty patients (19 males and 11 females) with ESRD undergoing dialysis, aged 22 to 64 years (mean+/-SEM: 49.1.0+/-1.9 years) were included in the study. The control group consisted of 20 healthy volunteers (13 males and 7 females) aged 35 to 55 years (mean+/-SEM: 46.2+/-1.4 years) matched according to sex and age. Blood samples were collected on the day preceding hemodialysis and one day following dialysis at 08:00, 12:00, 16:00, 20:00, 24:00, 02:00, 04:00, and 08:00 h. Melatonin concentration was measured by enzyme immunoassay. RESULTS: In patients with renal insufficiency undergoing dialysis mean melatonin nocturnal concentrations were significantly lower then those in healthy volunteers. The presence of the circadian rhythm in melatonin concentrations (although of significantly lower nocturnal amplitude) was detected only in 8 patients with renal failure undergoing hemodialysis, whereas in remaining 22 patients no such rhythm was found. Hemodialysis did not influence melatonin concentrations. CONCLUSIONS: The mechanism of depressed melatonin concentrations in hemodialyzed patients observed in our study remains unclear. However, it seems possible that decline in melatonin levels is due to impairment in adrenergic function that occurs in renal failure. Because the studies on the melatonin secretion in chronic renal failure bring about conflicting results, the relationship between renal diseases and melatonin secretion needs further investigations. PMID- 16380698 TI - Exercise intensity and its effects on thyroid hormones. AB - BACKGROUND: Physical activity influences energy metabolism in human subjects by increasing activity-induced energy expenditure and resting metabolic rate for several hours after exercise. Effects of exercise on circulating thyroid hormone values remain controversial. We have investigated the effect of acute aerobic exercise on thyroid hormone values. MATERIALS/METHODS: The effect of different intensity levels of acute aerobic exercise on thyroid hormones was investigated in 60 male well-trained athletes by performing bicycle ergometer at 45% (low intensity), 70% (moderate intensity), and 90% (high intensity). These intensities were selected according to their maximum heart rate (MHR). At each intensity level, heart rate, blood lactic acid, serum total thyroxine (T4), free thyroxine (fT4), total triiodothyronine (T3), free triiodothyronine (fT3) and thyroid stimulating hormone (TSH) values were measured. RESULTS: The results of this study show that exercise performed at the anaerobic threshold (70% of maximum heart rate, lactate level 4.59 +/- 1.75 mmol/l) caused the most prominent changes in the amount of any hormone values. While the rate of T4, fT4, and TSH continued to rise at 90% of maximum heart rate, the rate of T3 and fT3 started to fall. CONCLUSIONS: Maximal aerobic exercise greatly affects the level of circulating thyroid hormones. PMID- 16380699 TI - Shape characteristics of the foot arch: dynamics in the pregnancy period. AB - OBJECTIVES: The aim is data detection and finding some load consequences generated by various mechanical or physiological changes in the interaction of the end segment of the body--the foot--and the environment. DESIGN: Shape instability of the foot caused by e.g. loading of the foot by long-term frequency loads--walking, by extreme loads--sport, by hormonal changes--pregnancy, by aging, by pathologies, etc. The footprint surface was numerically described in 3D by means of stereo-photo-gram-metrical method--DMR digital relief model. Density of discrete points--250-400 per one print. SETTING: Detailed DMR was constructed by means of triangular web including contour picture with the use of Atlas program. The specified generated web is characterized by triangles with a cca 1 mm side in the number of up to 4,500 elements per one footprint model. RESULTS: The results enable us to deduce shape characteristics of DMR--the shape of the interactive boundary of the foot--the rest surface, to solve foot arch straining, to solve issues of discomfort and distribution of the pressure at the boundary of the foot--the rest surface, the shoe, etc. The gained findings can be interpreted in the field of prevention, therapy, orthopedics, podology, and enable us to come up with recommendations for the orthopedic practice and industrial use in the footwear production, etc. THE MAIN FINDINGS: The difference between volume reductions of the space under the foot arch characterizes the level of "fall" of the arch. This criterion is independent of the foot size, and is in 3D. CONCLUSIONS: Shape characteristics of footprints in pregnant women and in the period after childbirth were calculated on the basis of the defined criterion. The results of the group of four women tested in three periods suggest that there is no clear tendency towards the foot arch falling/increasing of the foot arch "fall" during the pregnancy period. PMID- 16380700 TI - Homocysteine levels in acromegaly patients. AB - Acromegaly is associated with a two to three-fold increase in mortality related predominantly to cardiovascular disease. The excess mortality is associated most closely with higher levels of growth hormone (GH). Survival in acromegaly may be normalized to a control age-matched rate by controlling GH levels; in particular, GH levels less than 2.5 ng/mL are associated with survival rates equal to those of the general population. Hyperhomocysteinemia has also been recognized as a risk factor for cardiovascular disease, yet there are limited data on the prevalence of hyperhomocysteinemia in patients with acromegaly. Eighteen acromegaly patients (7 male, 11 female, mean age 42.8 +/- 11.0 years) in our endocrine clinic consented to having the following tests performed: complete blood count (CBC), thyroid hormones, folic acid, vitamin B12, plasma homocysteine levels, uric acid, fibrinogen, CRP, fasting glucose, insulin, C-peptide, total serum cholesterol, HDL cholesterol, LDL cholesterol, triglycerides, GH, insulin like growth factor-1 (IGF-1) and GH levels after an oral glucose tolerance test (OGTT). By history, fourteen had macroadenomas and four had microadenomas; eight had hypertension; two had glucose intolerance, and four had diabetes. Fifteen had had transsphenoidal or transfrontal surgery: two had been cured, but 13 others were taking long-acting octreotide. Five patients had undergone radiotherapy and the acromegaly in two was treated primarily with long-acting octreotide. CBC, thyroid hormone, folic acid, and vit B12 levels were normal in all patients. We divided the patients into two groups according to mean GH levels after an OGTT: Group 1 (GH<2.5 ng/mL, n=10), and Group 2 (GH<2.5 ng/mL, n=8). Comparison of the two groups using Mann-Whitney U testing revealed statistically significant lower levels in Group 1 of the following parameters: GH (1.91 +/- 0.90 vs. 8.58 +/- 5.55 ng/mL, p=0.002), IGF-1 (338.30 +/- 217.90 vs. 509.60 +/- 293.58 ng/dL, p=0.06), GH after an OGTT (1.42 +/- 0.81 vs. 9.01 +/- 4.53 ng/mL, p=0.001), plasma homocysteine (12.85 +/- 4.47 vs. 18.20 +/- 4.99 micromol/L, p=0.05), total cholesterol (164.0 +/- 20.81 vs. 188.0 +/- 22.26 mg/dL, p=0.05) and LDL cholesterol (81.0 +/- 9.64 vs. 116.70 +/- 13.03 mg/dl, p=0.01). Differences between the other parameters were not significantly different. Acromegaly patients with high GH levels after an OGTT have much higher levels of homocysteine than patients with lower GH levels. The role of elevated homocysteine levels as an independent cardiovascular risk factor in the mortality of acromegaly patients should be determined in future studies. PMID- 16380701 TI - Circulating leptin, zinc, and copper levels after extracorporeal circulation. AB - OBJECTIVE: The role of leptin in the acute stress response to extracorporeal circulation has been well documented, however, the relationship between leptin and zinc has not been investigated previously. We aimed to research the circulating leptin, zinc, and copper levels before, during, and after the extracorporeal circulation, and effect of preoperative zinc administration to these. METHODS: Twenty patients who were taken to elective coronary artery bypass grafting operations using extracorporeal circulation were taken to this research and divided into two equal groups (n1, n2). In both groups blood samples were taken just before the operation (T0), at the end of operation (T1), and at the first postoperative day (T2). In the second group (n2) oral zinc (50 mg, once a day) was administered to patients for 5 days, preoperatively. The serum leptin, zinc, and copper levels were studied. RESULTS: In group n1 circulating leptin levels were significantly increased at T2 when compared to T0 and T1 (p<0.05); zinc levels were decreased at T2 when compared to T0 and T1 (p<0.05); copper levels were decreased at T2 when compared to T0 (p<0.05), and decreased at T1 when compared to T0 (p<0.05). In group n2 circulating leptin levels were significantly increased at T2 when compared to T0 and T1 (p<0.05); zinc levels were decreased at T2 when compared to T0 and T1 (p<0.05); copper levels were increased at T2 when compared to T1 (p<0.05). CONCLUSIONS: These results indicate that circulating leptin levels increase after the extracorporeal circulation as an acute response, while zinc and copper levels decrease at the same period. Preoperative zinc administration does not prevent the leptin response after extracorporeal circulation. PMID- 16380702 TI - Assessment of the influence of exercise on heart rate variability in anxiety patients. AB - OBJECTIVES: The aim of the study was to determine the status of the autonomic nervous system (ANS) in anxiety disorder patients and to evaluate possible exercise intervention in order to improve the ANS and overall psychiatric status of patients. PATIENTS AND METHODS: The ANS function was monitored via means of heart rate variability (HRV) changes during a 6-week hospitalization at the Psychiatric Clinic of University Hospital in Olomouc. The status of ANS was monitored by the new evaluation method of spectral analysis (SA) of HRV. The research involved 43 anxiety patients (29 women and 14 men) who underwent six weeks of intensive psychotherapy; part of the test group also participated in regular aerobic exercise. RESULTS: In the sense of autonomic dysfunction we can not give clear-cut answers to the question whether the autonomic dysfunction could be one of the predictors of anxiety disorder, although our results suggest lower vagal representation in the spectra. Unlike in the healthy population the exercise had a positive impact only on a limited number of patients. Regular endurance exercise on a stationary bicycle had a positive impact on the ANS efficiency only in patients with primary ANS activity reduction. In patients with normal ANS efficiency the exercise intervention had no effect; in some cases we even found activity reduction during the hospitalization period. However, the exercise had a positive effect on the course and outcome of the applied therapy in all patients. CONCLUSIONS: ANS disturbances have not been exhibited in some patients suffering from anxiety disorder. Due to the positive impact of regular physical activity on cardiovascular, metabolic, neural, and psychological changes in organism, monitored exercise should be implemented into therapeutic programs for patients with anxiety disorders. PMID- 16380703 TI - Somatotrophin axis hormones in patients affected with psoriasis. AB - OBJECTIVES: The serum concentrations of somatotrophin axis hormones (growth hormone--GH, insulin-like growth factor-I--IGF-I and insulin-like growth factor binding protein-3--IGFBP-3) in the patients with psoriasis in its active stage have been evaluated in relation to the control group consisting of healthy people in order to see whether these hormones may play a role in the psoriasis aetiopathogenesis. MATERIAL AND METHODS: The subjects included 85 psoriatic patients (mean age 36.9 +/- 11.2 years) in the active state of disease and 20 healthy persons (mean age 35.6 +/- 6.6 years) as a control group. GH and IGFBP-3 concentrations in serum were determined by immunoradiometric assay (IRMA), IGF-I concentrations--by radioimmunological assay (RIA). Evaluation of the dermatological state also included a determination of psoriasis area and severity index (PASI). RESULTS: In the patients with psoriasis the concentration of the growth hormone was found to be significantly higher than in the healthy people at the simultaneously much lower concentrations of the insulin-like growth factor-I and its binding protein-3. IGF-I concentrations correlated negatively to PASI value. CONCLUSION: In the psoriatic patients the somatotrophin axis activity is disturbed and these disturbances may effect the psoriasis process modulation. However, it is impossible to determine if the disturbances are of the primary, i.e. aetiological, importance for the disease pathogenesis, or only of the secondary--psoroid character. PMID- 16380704 TI - Patients with multiple sclerosis have higher levels of serum ghrelin. AB - In addition to metabolic and neuroendocrine actions, the recently discovered hormone ghrelin has been found to have inhibitory effects on inflammatory processes. This novel finding suggests possible involvement of the peptide in the pathogenesis of inflammatory disorders including the inflammatory demyelinating disease of the central nervous system, multiple sclerosis (MS). The aim of the present study was to evaluate serum ghrelin levels in patients with MS. Serum ghrelin levels were measured in 40 MS patients and 20 controls. Control subjects were selected from healthy individuals, matched for age, sex and BMI. Fasting plasma levels of ghrelin were determined by radioimmunoassay. Serum ghrelin level was significantly higher in MS group (226.16 +/- 35.84 pg/ml, n=40) than that in the control group (113.04 +/- 11.28 pg/ml, n=20, P<0.001). Both, relapsing remitting and secondary progressive MS patients had ghrelin levels significantly higher than controls, while there was no significant difference between the ghrelin levels of patients with these two categories of MS. This study for the first time shows that patients with MS have higher levels of ghrelin and this increase in circulating ghrelin level may function against the proinflammatory process in these patients. PMID- 16380705 TI - Does brand differentiate pharmaceuticals? AB - INTRODUCTION: Role of marketing in pharmaceutical industry is increasing and inspiration by successful brands known from consumer goods market influenced pharmaceutical companies enough to switch their attention to branding initiatives. Still there is little evidence that pharmaceutical brands represent anything more than product only. OBJECTIVES AND METHODS: This study aims to explore the area of branding in pharmaceutical industry. Central hypothesis of the research has been that brand and its emotional content differentiate pharmaceuticals as well as rational data derived from clinical studies. It has been tested by extensive review of available literature as well as by primary research focused on drivers of physicians' attitudes towards products and their influence on prescribing behavior. The research has been conducted in the sample of psychiatrists in the Czech Republic. RESULTS: No evidence about pharmaceutical brand exceeding value of product has been found in reviewed literature. Nevertheless, the primary research conducted in the sample of Czech psychiatrists indicates that emotional brand in pharmaceutical industry exists and enables author to draw a model of Customer/product life cycle that describes likely impact of functional, emotional and self-expressive benefits throughout pharmaceutical product's market presence. CONCLUSIONS: Pharmaceutical brand is likely to develop differently than the same of consumer goods products--it seems to be built predominantly on long-term positive experience. Marketing role in this process should lie in finding relevant product position and building brand identity compliant with real product capabilities. PMID- 16380706 TI - The circadian rhythm of tryptophan in breast milk affects the rhythms of 6 sulfatoxymelatonin and sleep in newborn. AB - INTRODUCTION: The hormone melatonin regulates the sleep and this pineal hormone is synthesized in the organism from the amino acid tryptophan. It is known that breast-fed babies have better sleep patterns and a better entrained sleep/wake cycle than bottle-fed babies (adapted formula). OBJECTIVE: To compare the circadian rhythm of 6-sulfatoxymelatonin (aMT6s)--the metabolite of melatonin excreted in the urine--in urine of bottle-fed and breast-fed children, and relate it to the circadian rhythm of tryptophan in breast milk, also evaluating the possible effects on the baby's night-time rest. METHODS: 16 infants of 12 weeks of age were studied, divided into two groups depending on their exclusively natural or artificial feeding. The circadian rhythm of 6-sulfatoxymelatonin in urine was measured for the two groups of infants and for the breast-feeding mothers. In the breast milk, the circadian rhythm of the amino acid tryptophan was measured. The rest of the infants was tested by wrist actimeters for a week and the sleep parameters of the infants were measured and evaluated. RESULTS: The tryptophan in the breast milk presented a circadian rhythm with acrophase at around 03:00. This affected the 6-sulfatoxymelatonin circadian rhythm with acrophase at 06:00 in the breast-fed infants, and also promoted nocturnal sleep. Assumed sleep, actual sleep, and sleep efficiency were significantly increased in the breast fed infants with respect the formula fed infants. CONCLUSION: A temporal relationship was observed between the circadian rhythm of 6 sulfatoxymelatonin of the exclusively breast-fed babies and that of tryptophan in the mother's milk. Acting this amino acid as a zeitgeber entrainment of the biological rhythms in the breast-fed infant. PMID- 16380707 TI - Effect of amylin on prolactin release. AB - OBJECTIVE: Amylin is a 37-amino acid peptide that is secreted from the pancreatic beta cells. This peptide is cosecreted with insulin from the pancreas by glucose. Amylin plays a role in glucose homeostasis and in the regulation of lipid metabolism. Amylin receptors were identified in central nervous system of rats. There is no available data on the effects of amylin on the pituitary hormones secretion. The aim of this study was to evaluate the effect of amylin on prolactin release in vivo and in vitro experiments in male adult Wistar Kyoto rats. METHODS: Amylin in a dose of 1 microg/5 microl was injected intraventricularly (i.c.v) during 5 min. using automatic pump. The control group received 5 microl cerebrospinal fluid. Amylin was injected intravenously (i.v) in a dose of 10 microg in 300 microl 0.9% NaCl. The control group received 300 microl 0.9% NaCl. In vitro experiment was performed in the pituitary cells culture conditions. Amylin was added to pituitary cell culture in a dose of 1, 10, 100 nM. Prolactin concentrations were determined using RIA methods. RESULTS: Central or peripheral administration of amylin caused a significant decrease of serum PRL concentrations as compared with control after 120 min. After 240 minute incubation of the pituitary cells culture with amylin in doses of 10 nM and 100 nM a significant inhibition of the release of PRL was found. However we found that the effect of amylin on the release of PRL depended on dose and time of incubation. A significant increase of PRL level was observed in cultured media in the presence of 1 nM of amylin after 60 min. CONCLUSION: Our results indicate that amylin administrated centrally and peripherally as well in the cell culture inhibits PRL release. PMID- 16380708 TI - Sibutramine therapy in obese women--effects on plasma neuropeptide Y (NPY), insulin, leptin and beta-endorphin concentrations. AB - OBJECTIVE: Some neuropeptides and monoaminergic neurotransmitters may affect hypothalamic feeding centres, sympathetic activity and thermogenesis. Sibutramine (BTS54524; N-[1-[1(4-chloro phenyl) cyclobutyl]-3methyl N,N-dimethylamine hydrochloride monohydrate) is a new 5-HT serotonin and noradrenaline reuptake inhibitor (SNRI), antiobesity drug. The aim of this study was to evaluate the effects of the sibutramine therapy on plasma neuropeptide Y (NPY), insulin, leptin and beta-endorphin concentrations in obese patients. METHODS: Sibutramine, serotonin and noradrenaline reuptake antiobesity drug was administered for 6 months in a dose of 10 mg daily in 60 obese women (BMI 30-40 kg/m2) (mean 34 kg/m2). Plasma NPY, leptin, beta-endorphin and insulin concentrations were measured with RIA methods using commercial kits (Peninsula Lab, Linco, Peninsula Lab, Swierk respectively). The above neuropeptides levels were evaluated before and after the 6 month sibutramine therapy in 60 obese women as well as in 30 obese women on low caloric diet and in 30 of the control group. RESULTS: In 85% obese patients a decrease of body weight was found after 6 month therapy with sibutramine. A decrease in total cholesterol, LDL and triglycerides and an increase in HDL were observed after the sibutramine treatment. We have demonstrated that the sibutramine therapy leads to the decrease of plasma NPY, beta-endorphin, insulin and leptin concentrations in obese patients. After low diet therapy we have observed a decrease in plasma leptin levels, however we did not find significant changes in plasma leptin, NPY, beta-endorphin and insulin concentrations. CONCLUSIONS: We suggest that the effects on the disturbed activity of NPY, beta-endorphin, insulin and leptin may be involved in the mechanism of sibutramine action. PMID- 16380709 TI - Hypertriglyceridemia and peripheral neuropathy in neurologically asymptomatic patients. AB - OBJECTIVES: In this study we intended to find a correlation between hypertriglyceridemia and peripheral neuropathy (PN) in patients with a high triglyceride level and no neurological complaints. METHODS: We recruited 24 patients (21 males and 3 females) having a triglyceride level above 300 mg/dl with no neurologic complaints and none of the other common causes of PN and they underwent an electroneurographic study. The distal motor or sensory latencies (DL), motor or sensory conduction velocities (CV), and motor or sensory amplitudes (AMP) were collected for the peroneal, posterior tibial, sural, median, and ulnar nerves and were considered abnormal if they fall above or below 2 standard deviations of reference values. RESULTS: Our results show that 70.8% of the patients had a significant delay in the DL of the sural nerves and 66.7% had a significant delay in the DL of the median sensory fibers. 54.2% of the patients had a significant decrease in the motor CV in the posterior tibial nerves and 33.3% had a significant decrease in the sensory CV in the sural nerves. The means of the DL and CV were significantly different from reference values in most of the nerves. Amplitudes were the least if at all affected. CONCLUSION: The pattern of the abnormalities affecting more the DL, the sensory nerves and the longer nerves of the lower extremities is suggestive of an early axonal polyneuropathy. We conclude that hypertriglyceridemia affects conduction parameters in peripheral nerves in a trend suggestive of early peripheral neuropathy. PMID- 16380711 TI - The role of microRNA-1 and microRNA-133 in skeletal muscle proliferation and differentiation. AB - Understanding the molecular mechanisms that regulate cellular proliferation and differentiation is a central theme of developmental biology. MicroRNAs (miRNAs) are a class of regulatory RNAs of approximately 22 nucleotides that post transcriptionally regulate gene expression. Increasing evidence points to the potential role of miRNAs in various biological processes. Here we show that miRNA 1 (miR-1) and miRNA-133 (miR-133), which are clustered on the same chromosomal loci, are transcribed together in a tissue-specific manner during development. miR-1 and miR-133 have distinct roles in modulating skeletal muscle proliferation and differentiation in cultured myoblasts in vitro and in Xenopus laevis embryos in vivo. miR-1 promotes myogenesis by targeting histone deacetylase 4 (HDAC4), a transcriptional repressor of muscle gene expression. By contrast, miR-133 enhances myoblast proliferation by repressing serum response factor (SRF). Our results show that two mature miRNAs, derived from the same miRNA polycistron and transcribed together, can carry out distinct biological functions. Together, our studies suggest a molecular mechanism in which miRNAs participate in transcriptional circuits that control skeletal muscle gene expression and embryonic development. PMID- 16380712 TI - Identification of direct DAF-16 targets controlling longevity, metabolism and diapause by chromatin immunoprecipitation. AB - DAF-16, a forkhead transcription factor, is a key regulator of longevity, metabolism and dauer diapause in Caenorhabditis elegans. The precise mechanism by which DAF-16 regulates multiple functions, however, is poorly understood. Here, we used chromatin immunoprecipitation (ChIP) to identify direct targets of DAF 16. We cloned 103 target sequences containing consensus DAF-16 binding sites and selected 33 targets for further analysis. Expression of most of these genes is regulated in a DAF-16-dependent manner, and inactivation of more than half of these genes significantly altered DAF-16-dependent functions, including life span, fat storage and dauer formation. Our results show that the ChIP-based cloning strategy leads to greater enrichment for DAF-16 target genes than previous screening strategies. We also demonstrate that DAF-16 is recruited to multiple promoters to coordinate regulation of its downstream targets. The large number of target genes discovered provides insight into how DAF-16 controls diverse biological functions. PMID- 16380713 TI - Trak1 mutation disrupts GABA(A) receptor homeostasis in hypertonic mice. AB - Hypertonia, which results from motor pathway defects in the central nervous system (CNS), is observed in numerous neurological conditions, including cerebral palsy, stroke, spinal cord injury, stiff-person syndrome, spastic paraplegia, dystonia and Parkinson disease. Mice with mutation in the hypertonic (hyrt) gene exhibit severe hypertonia as their primary symptom. Here we show that hyrt mutant mice have much lower levels of gamma-aminobutyric acid type A (GABA(A)) receptors in their CNS, particularly the lower motor neurons, than do wild-type mice, indicating that the hypertonicity of the mutants is likely to be caused by deficits in GABA-mediated motor neuron inhibition. We cloned the responsible gene, trafficking protein, kinesin binding 1 (Trak1), and showed that its protein product interacts with GABA(A) receptors. Our data implicate Trak1 as a crucial regulator of GABA(A) receptor homeostasis and underscore the importance of hyrt mice as a model for studying the molecular etiology of hypertonia associated with human neurological diseases. PMID- 16380714 TI - Conserved noncoding sequences are selectively constrained and not mutation cold spots. AB - Noncoding genetic variants are likely to influence human biology and disease, but recognizing functional noncoding variants is difficult. Approximately 3% of noncoding sequence is conserved among distantly related mammals, suggesting that these evolutionarily conserved noncoding regions (CNCs) are selectively constrained and contain functional variation. However, CNCs could also merely represent regions with lower local mutation rates. Here we address this issue and show that CNCs are selectively constrained in humans by analyzing HapMap genotype data. Specifically, new (derived) alleles of SNPs within CNCs are rarer than new alleles in nonconserved regions (P = 3 x 10(-18)), indicating that evolutionary pressure has suppressed CNC-derived allele frequencies. Intronic CNCs and CNCs near genes show greater allele frequency shifts, with magnitudes comparable to those for missense variants. Thus, conserved noncoding variants are more likely to be functional. Allele frequency distributions highlight selectively constrained genomic regions that should be intensively surveyed for functionally important variation. PMID- 16380715 TI - Cooperative and antagonistic interactions between Sall4 and Tbx5 pattern the mouse limb and heart. AB - Human mutations in TBX5, a gene encoding a T-box transcription factor, and SALL4, a gene encoding a zinc-finger transcription factor, cause similar upper limb and heart defects. Here we show that Tbx5 regulates Sall4 expression in the developing mouse forelimb and heart; mice heterozygous for a gene trap allele of Sall4 show limb and heart defects that model human disease. Tbx5 and Sall4 interact both positively and negatively to finely regulate patterning and morphogenesis of the anterior forelimb and heart. Thus, a positive and negative feed-forward circuit between Tbx5 and Sall4 ensures precise patterning of embryonic limb and heart and provides a unifying mechanism for heart/hand syndromes. PMID- 16380716 TI - A unified mixed-model method for association mapping that accounts for multiple levels of relatedness. AB - As population structure can result in spurious associations, it has constrained the use of association studies in human and plant genetics. Association mapping, however, holds great promise if true signals of functional association can be separated from the vast number of false signals generated by population structure. We have developed a unified mixed-model approach to account for multiple levels of relatedness simultaneously as detected by random genetic markers. We applied this new approach to two samples: a family-based sample of 14 human families, for quantitative gene expression dissection, and a sample of 277 diverse maize inbred lines with complex familial relationships and population structure, for quantitative trait dissection. Our method demonstrates improved control of both type I and type II error rates over other methods. As this new method crosses the boundary between family-based and structured association samples, it provides a powerful complement to currently available methods for association mapping. PMID- 16380718 TI - Embracing risk. PMID- 16380717 TI - In vivo imaging of islet transplantation. AB - Type 1 diabetes mellitus is characterized by the selective destruction of insulin producing beta cells, which leads to a deficiency in insulin secretion and, as a result, to hyperglycemia. At present, transplantation of pancreatic islets is an emerging and promising clinical modality, which can render individuals with type 1 diabetes insulin independent without increasing the incidence of hypoglycemic events. To monitor transplantation efficiency and graft survival, reliable noninvasive imaging methods are needed. If such methods were introduced into the clinic, essential information could be obtained repeatedly and noninvasively. Here we report on the in vivo detection of transplanted human pancreatic islets using magnetic resonance imaging (MRI) that allowed noninvasive monitoring of islet grafts in diabetic mice in real time. We anticipate that the information obtained in this study would ultimately result in the ability to detect and monitor islet engraftment in humans, which would greatly aid the clinical management of this disease. PMID- 16380720 TI - Widening the spectrum of human genetic variation. PMID- 16380721 TI - Amyloid double trouble. PMID- 16380722 TI - Beyond the beta cell in diabetes. PMID- 16380723 TI - Confronting ethnicity-specific disease risk. PMID- 16380727 TI - Surprising shrinkage of expanding gels under an external load. AB - Hydrogels are fascinating and useful in that they can show large volume changes in response to various stimuli, such as temperature or chemical environment. Here we report the peculiar observation that chemically crosslinked hydrogels that normally expand owing to a change in electrolyte pH can be made to shrink in certain circumstances. Specifically, these hydrogels contract when tested at a constant compressive force and subjected to a pH change that causes expansion in the absence of the applied load. When tested under tension, the gels always expand. Although the effects of external stress on the swelling of gels is known, the concomitant change in gel mechanical properties during pH switching was found to be a more dominant effect in our studies. However, existing mechanical models used to predict dimensional changes in actuator materials could not explain both the tensile and compression results. In addition, we show that the friction between metal plates of the apparatus and the gel is a key factor in explaining the contractile actuation under compressive loads. The observations reported in this paper are important for the successful design and use of hydrogel actuators in devices such as valves for microfluidics. PMID- 16380729 TI - Study of colloidal quantum-dot surfaces using an innovative thin-film positron 2D ACAR method. AB - Nanosized inorganic particles are of great interest because their electronic properties can be easily tailored, providing a tremendous potential for applications in optoelectronic devices, light-emitting diodes, solar cells and hydrogen storage. Confinement of electrons and holes to dimensions comparable to their wavelength leads to quantum-well states with modified wavefunctions and density of states. Surface phenomena are crucial in determining nanoparticle properties in view of their large surface-to-volume ratio. Despite a wealth of information, many fundamental questions about the nature of the surface and its relationship with the electronic structure remain unsolved. Ab initio calculations on CdSe nanocrystals suggest that passivating the ligands does not produce the ideal wurtzite structure and that Se atoms relax outwards irrespective of passivation. Here we show that implanted positrons are trapped at the surface of CdSe nanocrystals. They annihilate mostly with the Se electrons, monitor changes in composition and structure of the surface while hardly sensing the ligand molecules, and we thus unambiguously confirm the predicted strong surface relaxation. PMID- 16380730 TI - Atomistic simulations of spinodal phase separation preceding polymer crystallization. AB - Many polymeric materials crystallize when cooled below their melting temperature. Although progress has been made in our understanding of the crystallization process through both experimental and theoretical efforts, these studies have focused mainly on the crystal nucleation and growth mechanism, where critical nuclei are formed from a metastable state during the first stages of crystallization, leading ultimately to the growth of crystal domains. Attention has also been given to the structure during the precrystallization (induction period). A pretransition state occurring before crystallization has been characterized as an unstable phase separation initiated by density and orientational fluctuations. These fluctuations are caused by an increase in the average length of rigid trans segments along the polymer backbone during the induction period. These observations are consistent with the theory proposed in ref. 14 on the isotropic-to-nematic transition of polymer liquid crystals, that is, the parallel ordering of polymers is caused by an increase in chain rigidity. Here we use large-scale computer simulations to investigate melts of polymers in the early ordering stages (induction period) before crystallization. In the ordered domains we identify growing dense regions similar to smectic liquid crystals. Our simulations reveal a 'coexistence period' in the ordering before crystallization, where nucleation and growth mechanisms coexist with a phase separation mechanism. PMID- 16380731 TI - Ocular morbidity from sickle cell disease in a Nigerian cohort. AB - Sickle cell Retinopathy is increasingly being recognised as a cause of significant ocular morbidity and blindness in Africa south of the Sahara. This study looked for retinopathy in a cohort of 90 Nigerians with Sickle Cell Disease (SCD). METHOD: The cohort consisted of black Nigerians from the Hausa-Fulani, Ibo and Yoruba, as well as other minority ethnic groups resident in the Federal Capital Territory aged between 5-36 yr. 88 patients were SS and only 2 SC. RESULTS: SCD related posterior lesions were seen in 22 patients (24%). Of these, 19 cases (21%) had Non Proliferative Sickle Retinopathy (NPSR) while 5 (5.6%) had Proliferative Sickle Retinopathy (PSR) in various stages of development, and 2 had both PSR and NPSR. Patients with PSR are at risk of blindness from vitreous haemorrhage and tractional retinal detachment. A 14-year-old male with arterio venous anastomosis was the youngest with PSR while the most advanced PSR lesion was a sea fan in a 25-year-old female. CONCLUSIONS: standard treatment consisting of photocoagulation and/or vitrectomy is not available in many eye centres in sub Saharan Africa and steps need to be taken to improve this situation. The role of anti-sickling remedies, if any, is the subject of ongoing investigations. Our findings with NIPRISAN, a phytomedicinal preparation currently undergoing trials, will be reported subsequently. PMID- 16380732 TI - Clinical features of goitres on the Nigerian Plateau. AB - To describe the prevalence, clinical types and aetiology of goitre among hospital patients on the Nigeria highland area of Plateau. Subjects, materials and methodology: Adult out-patients attending the medical, surgical and general outpatient clinics of the Jos University Teaching Hospital (JUTH) were prospectively examined for the presence of goitre. Goitrous patients were subjected to further evaluation. Goitre was graded according to the WHO (1979) criteria (grades 0 - 4) and characterised as to lobarity, nodularity, consistency, tenderness, mobility of overlying skin and presence of bruit as well as hormonal functional status. RESULTS: 3,000 outpatients were examined, 124 (4.1%) of which were goitrous. Of the goitrous patients 98 (84F, 14M) had full data to be eligible for further analysis. The goitrous patients were distributed according to goitre grade as follows: grade one 10 (10.2%), grade two 62 (63.3%), grade three 22 (22.5%) and grade four 4 (4.1%). Both right and left lobes were enlarged in 91 (92.9%) subjects. Enlargement was limited to the right lobe in one patient with 3 (3.1%) patients each showing goitre limited to the left lobe and the isthmus. Palpably nodular goitre was found in 21 (21.4%) patients while the large majority of 78.6% showed a diffuse enlargement. The goitre was firm in 55% of cases and soft in 27.5% of patients. Goitre was associated with thyrotoxicosis in 10 (10.2%) patients, carcinoma in 2(2.1%), thyroid cyst in 2(2.1%) and thyroiditis in 6(6.1%) patients. CONCLUSION: Goitre is common among out-patients on the Nigerian plateau, with a characteristic female preponderance. WHO grade 2 goitre is the most common form. Thyrotoxicosis is relatively common among the goitrous patients. Patients attending hospital in goitre-endemic areas should be routinely examined for thyroid enlargement and those found goitrous subjected to endocrine evaluation. PMID- 16380733 TI - What impressions do medical students of the University of Benin have about anaesthesia and anaesthesiologists? AB - A survey on the impressions 500 level medical students of the University of Benin have about anaesthesia and anaesthesiologists was carried out from May to August 1999. The survey showed the mean age of the students to be 25.3 years and a male to female ratio of 3:1. The study also showed that 170 (82.5%) of the students were unaware of the scope of anaesthesia before the posting. After the posting, this figure decreased to 58 (34.5%) (chi2 = 11.7, p < 0.05, df = 2). Approximately same number of females (71.1%) and males (72.3%) had low awareness (unaware and limited awareness) about the scope of anaesthesia. However, more males (16.9%) were fully aware about anaesthesia and anaesthesiologists than female students (9.4%). Similarly, 76 (54.3%) of the students knew that anaethesiologists were doctors before the posting. This figure improved to 109 (85.2%) after the posting and was statistically significant. Interest in the specialty was exhibited by only 2 (2%) of the students before the posting and 21 (25%) of them became interested during and 4 (5%), after the posting in anaesthesia. The impressions had by those uninterested in anaesthesia include the unchallenging, non-lucrative and stereotyped nature of the specialty. It also included the dependency of the specialty on other specialties and that it attracted dull, lazy doctors whose job was unrecognized. This study's findings suggest the need for better exposure of medical students to anaesthesia to increase awareness and for the specialty to grow. KEYWORDS: medical students, awareness, anaesthesia, anaesthesiologists. PMID- 16380734 TI - Knowledge and practice of cervical screening among female health workers in Sokoto, North Western Nigeria. AB - BACKGROUND: Several studies have shown the importance of health care professionals as predictors of the use of cervical cancer screening. AIMS: To determine the knowledge and utilisation of cervical cancer screening services in a cohort of subjects where awareness is naturally expected to be high. METHODS: A cross sectional questionnaire survey of 159 female health care providers in Usmanu Danfodiyo University Teaching Hospital Sokoto. The sample included doctors, nurses, pharmacists, laboratory scientists and medical social workers. RESULTS: Knowledge of cervical cancer screening is high among the respondents. However, only 4.4 % had availed themselves of the opportunity for the test. CONCLUSION: Health care providers should improve on opportunistic screening. It is necessary for a national screening policy to be formulated and implemented in Nigeria. PMID- 16380736 TI - Sexual behaviour, perception of HIV/AIDS and condom use among commercial motorcylists in Benin City. AB - OBJECTIVE: This study aims to evaluate the perception of HIV/AIDS, condom use and the sexual behaviour of commercial motorcyclists in Benin-city, Edo State. DESIGN: A questionnaire based study involving consecutive 160 male commercial motorcyclists. SETTING: The study was conducted in three randomly selected major motorcycle parks in Benin-city. The three parks were selected using a multi stage random sampling technique. In each of the selected parks, consecutive motorcyclists were interviewed until a total of 160 respondents had been interviewed. RESULTS: One hundred and fifty-two (95%) of the respondents were acquainted with the subject of HIV/AIDS, but only 140(87%) believed that the disease actually existed. Sixty-six per cent of the respondents had multiple sexual partners while 26% of the subjects had been treated for a sexually transmitted infection. Of the 54 (34%) men who believed the disease had a cure, 46% and 32% felt that the cure was herbal and spiritual respectively. Although 106 of the men (66%) acknowledged the preventive function of the condom in the spread of HIV/AIDS and other sexually transmitted infections, only 45% used the condom regularly with sexual partners. Sixty-two per cent of the respondents would submit themselves to voluntary HIV screening though none had gone for voluntary screening. CONCLUSION: The study suggests that the sexual behaviour of the commercial motorcyclist and a scarcity of information expose him to increased risk of HIV/AIDS infection and other sexually transmitted infections. PMID- 16380735 TI - Knowledge and perception of obstetric analgesia among prospective parturients at the Lagos University Teaching Hospital. AB - BACKGROUND: Natural labour is painful. Such pain has little value and is potentially harmful to the parturient and foetus. OBJECTIVE: To assess knowledge, perceptions and acceptance of obstetric analgesia among prospective parturients. The effect of age, educational status and parity on the above variables was evaluated. DESIGN AND SETTING: Prospective cross-sectional study between October 2002 and April 2003 at the Lagos University Teaching Hospital (LUTH). PATIENTS AND METHODS: Patients attending ante-natal clinics voluntarily responded to a structured questionnaire which enquired patient's age, educational status and parity; knowledge, perceptions and acceptance of obstetric analgesia. The association between the variables was determined with the Pearson's Chi square test. RESULT: Four hundred and fifty patients of varying educational levels were studied. Only 175 (38.9%) knew of obstetric analgesia. There was a significant association between educational status and knowledge of obstetric analgesia (p = 0.000). Two hundred and ninety four patients (65.3%) would accept analgesia during labour of which 179 (60.9%) would leave the choice to the doctor to give them the "best available". There was no association between age, educational status and parity with acceptance (p > 0.05). Reasons for rejection included labour being a 'natural process' in 15.1%, faith in divine intervention 6.0% and concerns about side effects 3.1%. CONCLUSION: This study has revealed a dearth of knowledge and gross misconception about obstetric analgesia. With proper education, many patients will accept some form of analgesia during labour. There is an urgent need to commence standard obstetric analgesia services in our tertiary hospitals. PMID- 16380737 TI - Sero-prevalence of hepatitis B surface antigen (HBsAg) in pregnant women in Port Harcourt, Nigeria. AB - BACKGROUND/OBJECTIVE: Hepatitis B Virus (HBV) infection is a global public health problem and is endemic in Asia and Sub-Saharan Africa, including Nigeria. Vertical transmission of HBV infection is thought to be a major mode of transmission in endemic areas. This study was therefore undertaken to determine the sero-prevalence of Hepatitis B surface antigen (HBsAg) on pregnant women in Port Harcourt, Nigeria, in an effort to obtain baseline information on disease burden in this locality, in order to institute public health measures to reduce transmission. PATIENTS AND METHODS: Serial recruitment of 600 women attending the ante-natal clinic at the University of Port Harcourt Teaching Hospital, Port Harcourt, Nigeria. HBsAg screening was done using a rapid ELISA Kit. RESULTS: 26(4.3%) of the 600 subjects tested positive to HBsAg. Blood transmission, dental manipulations, tattooing and circumcision were found not to be significant modes of transmission of HBV infection in Port Harcourt. CONCLUSION: There is an intermediate prevalence (4.3%) of HBV infection in pregnant women in Port Harcourt, Nigeria. The need to institute public health measures to reduce disease burden and transmission, including routine screening of all pregnant mothers for HBV infection and early passive/active immunisation of babies born to HBsAg- positive mothers is advocated. PMID- 16380738 TI - Haematological parameters in sickle cell anaemia patients with and without splenomegaly. AB - BACKGROUND: The aim is to investigate the clinical and haematologic effects of persistent splenomegaly (PS) and splenectomy in a population of Nigerians with sickle cell anaemia (SCA). MATERIALS AND METHODS: SCA patients attending the Haematology clinic of the OAUTHC, Ile-Ife, were examined physically and haematologically. Patients with PS were compared with age and sex-matched controls without splenomegaly. Foetal haemoglobin (HbF), packed cell volume (PCV), reticulocyte count and the frequency of blood transfusion were noted for both groups. The clinical effects of splenectomy were documented in three patients who underwent the procedure because of symptomatic massive splenomegaly (> or = 10 cm). RESULTS: Seventy-one patients, 40 males and 31 females, aged 16 48 (median, 21) years were studied. Nineteen (26.8%) had PS ranging from 3-22 cm (mean +/- SD = 9.4 +/- 4.4 cm; median = 7 cm). Ten of these had massive splenomegaly (spleen > or = 10 cm) varying from 10-22 cm (mean +/- SD = 12.3 +/- 3.8 cm; median = 12 cm). No significant differences were found between the mean values of HbF, PCV, reticulocyte count, frequencies of pain crisis, transfusion requirement, weights and heights in SCA with PS and age-sex matched controls without PS. Annual transfusion requirement showed a mild negative correlation with splenic size (r = -0.06), which was not statistically significant (p = 0.882). Significant haematologic improvement was confirmed in the 3 patients who underwent elective splenectomy for splenomegaly-induced hypersplenism. CONCLUSIONS: We conclude that massive persistent splenomegaly is a recognised cause of significant morbidity in SCA patients as evident in the patients that underwent splenectomy. However, where it is mild to moderate, PS may not have significant adverse effect on the overall clinical picture of the disorder. PMID- 16380739 TI - Pattern of cerebral malaria in children at the Lagos University Teaching Hospital. AB - Cerebral malaria is one of the most lethal forms of malaria. Given that malaria is a constantly evolving disease, it is therefore necessary to document patterns of presentation even in the same centre over a period of time. OBJECTIVE: To document the prevalence and pattern of cerebral malaria in children. DESIGN: Cross-sectional descriptive study of children with cerebral malaria attending the emergency room of the Lagos University Teaching Hospital. Age, sex, month at which diagnosis was made, associated clinical features, condition at discharge and mortality were assessed. RESULTS: Cerebral malaria was documented in 107/3309 (3.2%) children. There was an equal male:female ratio. Cerebral malaria occurred most frequently between July and September and in children between 2 and 2.9 years. A total of 79/107 (73.8) recovered fully at discharge, 9/107 (8.4%) recovered with some neurological sequelae while 19/107 (17.8%) died. Coma score on admission was significantly lower among those who died compared with those who survived (p = 0.001). Clinical signs observed in these children were seizures 88/107 (82.2%), pallor-75/107 (70.1%), jaundice-55/107 (51.4%) and hepato splenomegaly-18 (16.8%). However, decerebrate posturing was the only clinical sign associated with a fatal outcome OR, 11.47 (p = 0.009). CONCLUSION: This study shows that cerebral malaria still remains a problem of the under fives with unacceptably high mortality. The clinical significance of decerebrate posture as an indicator of mortality would require further evaluation. PMID- 16380740 TI - Improving attitude towards elderly people: Evaluation of an intervention programme for caregivers. AB - There is yet no specific Government policy on care of the elderly in Nigeria despite increase in its awareness as well as the number of people reaching old age. This study was carried out to assess the attitude of caregivers in seven out of the eleven wards in Ilorin metropolis. The caregivers consisted of daughters (44.4%), sons (18.3%), family-relation (24.6%), employees (1.4%), neighbors (4.5%) and community members (6.8%). The data collection toll was a structured questionnaire consisting of three parts designed to determine the socio demographic characteristics, general and specific attitude to the care of the elderly. Effect of Health Education on attitude of caregivers concerning the care of the elderly was also evaluated. Based on the scoring system, the attitude of the caregivers was found to be fairly positive and after the Health Education intervention, a significant improvement in the attitude of caregivers was recorded. The Mean (SD) score increased significantly from 78.4 +/- (14.6) pre intervention to 96.8 +/-(15.2), post-intervention, p-value < 0.05. Religious teaching (35.9%), traditional/customary obligation (28.7%) and personal feeling (21.1%) were the most influencing factors of attitude of caregivers towards the care of the elderly. The influence of educational status of caregivers on where the elderly should be taken care of was found to be non statistically significant (p > 0.05). Gender had overall positive effect on attitudinal measurement. The deduction from this study is that care of the elderly is still being valued despite lack of specific government policy and support. The negative attitude of caregivers to institutional care may be difficult to change through health education intervention because care of the elderly at home is still being greatly valued in this country as it is seen as a responsibility of children and other family members. Institutional care is also considered as an abomination and irresponsibility by some caregivers Keywords: attitude, caregivers, elderly, intervention and health education. PMID- 16380741 TI - Assault cases in Maiduguri metropolis: A comparative study of police and accident and emergency units' records. AB - BACKGROUND: Information on the incidence of assault cases such as armed robbery, fights and house burglaries in most Nigerian cities are usually based on data collected from police records with little or no attempt to tap medical sources of information. OBJECTIVE: The aim of this study was therefore to compare the rate of recording of assault cases in the Maiduguri metropolitan area by the police at the Maiduguri central police station and the Accident and Emergency unit (A and E) of the University of Maiduguri Teaching Hospital. Possible reasons why assault victims report or do not report to the police before seeking for treatment was also investigated. METHOD: All victims of assault who attended the A and E unit of the hospital over a one year period (2002-2003) were interviewed and relevant data collected in a prepared form. Findings were compared with the record of assault cases within the same period at the Maiduguri central police station. RESULT: 185 patients presented at A and E with body injuries due to assault during period of study. 108 (58%) informed the police before attending hospital, out of which majority (38.9%) claimed an 'instinctive reaction to do so' as the reason for informing the police. A significant proportion (61%) of those that did not inform the police claimed that the feeling that 'nothing will come out of it' as the reason. Only 41% of cases reported to the police were recorded. There was a tendency by the police to record cases where gun was used or the victim was a female. CONCLUSION: The true picture of violence in most Nigerian cities can better be assessed by collecting data from both the relevant A and E department as well as from the police. The need for the populace to develop confidence and trust in the police cannot be overemphasised. PMID- 16380742 TI - Helicobacter pylori and the pathogenesis of gastroduodenal disease: Implications for the management of peptic ulcer disease. AB - The discovery of Helicobacter pylori has changed our understanding of the pathophysiology of peptic ulcer disease. An estimated one billion people harbour the organism worldwide but the highest prevalence is found in developing countries with up to 80% of people infected. The most favoured modes of transmission are faeco-oral and oral-oral. The mechanisms of H pylori-induced gastroduodenal disease include the provocation of local inflammatory reaction with the release of toxic cytokines, elevation of gastrin concentration and cytotoxic epithelial injury from the activity of urease and other enzymes produced by the bacterium. However, a large proportion of infected persons have no disease or are asymptomatic thereby suggesting that there may be other factors apart from H pylori infection necessary for ulcer formation. The simple finger prick test, a variant of serology and the newly developed ELISA-based Faecal Antigen Test hold the ace for large-scale epidemiological studies. The eradication of H pylori is now a very important goal of treatment of gastric and duodenal ulcers. Most H pylori eradication regimens combine anti-secretory agent, usually a proton pump inhibitor or H2-receptor antagonist and two antibiotics (usually, Clarithromycin and Amoxycillin or Metronidazole). Emergence of antibiotic resistance is worrisome but a quadruple therapy that incorporates bismuth may be used if the triple therapy fails. PMID- 16380743 TI - Surgical lymph node biopsies in University of Ilorin Teaching Hospital, Ilorin, Nigeria. AB - BACKGROUND: To determine the diagnostic value of lymph-node biopsy, commonest causes of lymph node enlargement requiring biopsy and the usual nodes involved. METHODS: A retrospective study of 169 lymph node biopsies representing 97% of lymph node biopsies and 3.2% of total biopsies specimen received at Pathology Department of University of Ilorin Teaching Hospital Ilorin, Nigeria over a 5 year period was undertaken. RESULTS: Tuberculous lymphadenitis (38%) and metastatic nodal involvement (25%) were the commonest causes of lymph node enlargement. Generalised lymphadenopathy occurred in 105 (62%) patients while localised enlargement was seen in 64{38%) of cases. The commonest sites of localised lymphadenopathy were axillary (38%), cervical (32%), inguinal (8%), and submandibular (8%). While axillary lymph node enlargements were mostly associated with tumour metastasis, cervical node enlargements were mostly associated with tuberculosis. Supraclavicular lymphadenopathy has the highest risk of malignancy. CONCLUSION: The result shows that tuberculous lymphadenitis and metastatic nodal malignancies are the commonest causes of lymph node enlargement in our environment. Reasons for this include the often involvement of lymph nodes in tuberculous infections {as lymphadenitis constituted the most frequent form of extra pulmonary tuberculosis} and cancer's metastasis. With an effective and nationwide immunisation against tuberculosis, health education on HIV infection coupled with early screening and detection of malignancies in generals, we hope this trend will change in the future. Lymph node biopsy remains an important and valuable diagnostic tool in evaluation of lymph node enlargement as it allows for the architecture of the gland to be viewed thereby given an accurate and concise diagnosis with very minimal risk to the patient. PMID- 16380744 TI - The pregnant Rhesus negative Nigerian woman. AB - OBJECTIVE: Alloimmunisation to Rhesus D (RhD) is a major factor in perinatal morbidity and may result in the compromise of the woman's obstetric career. In Nigeria accurate population based studies to determine the prevalence of Rhesus negative women and the incidence of alloimmunisation are lacking, hence we undertook to study pregnancy outcome in Rhesus negative women. METHOD: We studied retrospectively sixty-seven RhD negative women over a two year period; information was obtained from the case-file of all pregnant women who presented to the ante-natal clinic and were identified as Rhesus negative. This was corroborated with the blood bank record over the same period. RESULT: Forty per cent of these were nulliparae with an average of 0.5 abortions. Only 20% had the blood groups of their husbands documented, and only four babies born to these women had their Rhesus group recorded. Six of the babies appeared to have been severely affected by Rhesus isoimmunisation. Three of these had an exchange blood transfusion (EBT); all who had an EBT had a satisfactory outcome. Out of the other three, there were two neonatal deaths and one fresh stillbirth. Fourteen babies had neonatal jaundice with a mean bilirubin level of 6 mg/dl, all of which were mainly unconjugated. Evidence for the administration of anti D was obtained in only three patients; all had one ampoule (dose in i.u and/or _g not stated) administered within 72 hr. CONCLUSION: This preliminary study has shown that isoimmunisation due to Rhesus incompatibility is poorly studied among Nigerian women with many questions unanswered; therefore there is an urgent need for a management protocol for this condition, which will include both the clinicians and the laboratory physicians. PMID- 16380745 TI - Cancer of the cervix. AB - Cases of cancer of the cervix histologically diagnosed at the Olabisi Onabanjo University Teaching Hospital, Sagamu, Nigeria between January 1991 and December 2000 were reviewed. There was 1,912 total gynaecological admissions during that period and 56 histologically diagnosed cancer of the cervix for an incidence of 2.9%. The mean age at occurrence of the cancer was 51.7 + 12.4 years. It was more common in grandmultiparous women and major presenting complaints were abnormal vaginal bleeding, (64.3%) watery discharge (25.0%) and postcoital bleeding (7.1%). Histologically, 96.4% has squamous cell carcinoma of varying degree of differentiation and most (78.6%) presented in the advanced stages. Majority (80.4) were referred for radiotherapy at either Lagos University Teaching Hospital or University College Hospital, Ibadan. Only 5.4% of cases were treated by radical hysterectomy. Measures to reduce the incidence and morbidity would include mass education of the sexually active women to have cervical smear regularly and also to report symptoms early so as to diagnose the invasive disease in the early stages. PMID- 16380746 TI - Surgical management of a large complex odontoma of the mandibular angle-ramus region through intra-oral buccal approach--A case report. AB - Large complex odontomas of the jaws are rare. A report of a large complex odontoma of the mandibular angle-ramus region enucleated through intra-oral buccal approach is presented. A review of the literature on different modalities of treatment is also undertaken. A large expansile complex odontoma of the angle ramus region of the mandible was excised through an intraoral buccal approach under general anaesthesia. Recovery and immediate post-operative period were uneventful. There was no altered sensation in the distribution of the inferior alveolar nerve and lingual nerve; and wound healing was satisfactory. Post operative radiograph 2 years after the operation showed satisfactory bone regeneration. Intraoral buccal approach to large complex odontomas of the angle ramus region of the mandible is a relatively safe procedure with minimal complication. PMID- 16380747 TI - [Neuropsychiatric systemic lupus erythematosus: where are we now?]. AB - When dealing with Neuropsychiatric Systemic Lupus Erythematosus (NPSLE) there are still many controversial topics. In 1999 the American College of Rheumatology gave classification criteria for 19 clinical syndromes. However major problems are still related to low specificity of some of them such as headache, cognitive impairment or mood disorders. Even though a frequency of CNS involvement from 14 to 75% has been described, depending on both the population studied and the methodology of assessment, a lower frequency ranging from 21 to 28 % derived by larger case series seems more realistic. The introduction of the concept of "borderline cases", proposed by Italian Study Group for NP-SLE, is based both on clinical and instrumental evaluation and could represent a useful tool when dealing with conditions which do not fulfil ACR classification. Also the relationship between SLE activity and NP involvement is a debated issue. Concerning pathogenesis, it seems reasonable to consider multifactorial mechanisms related to antibody-mediated damage, antiphospholipid pro-thrombotic effect, non-inflammatory vasculopathy and cytokines mediated cytotoxycity. However, direct and unequivocal evidence for the implication of any of the above mentioned mechanisms is still lacking. Although a wide range of neuroimaging tools have been used to evaluate CNS involvement, no single technique has proven to be definitive and, when dealing with a patient with suspected NPSLE, it is important to combine different diagnostic techniques. Due to the lack of effective imaging along with limitation in knowledge of underlying pathogenetic mechanisms and paucity of histopathologic findings, therapeutic approach in NPSLE remains a difficult issue and is currently based on personal experience. Italian Study Group for NP-SLE proposes the creation of a national registry on NPSLE to validate ACR classification criteria. Furthermore, the possibility to collect large series and stratifying them for each of the included neuro-psychiatric syndromes seems a good strategy for planning multicentric controlled therapeutic trials in the near future. PMID- 16380748 TI - [Complementary medicine in rheumatoid arthritis]. AB - Use of complementary and alternative medicine (CAM) for chronic conditions has increased in recent years. CAM is immensely popular for musculoskeletal conditions and patients suffering from rheumatoid arthritis (RA) frequently try CAM. This review summarises the trial data for or against CAM as a symptomatic treatment for rheumatoid arthritis. Collectively the evidence demonstrates that some CAM modalities show significant promise, e.g. acupuncture, diets, herbal medicine, homoeopathy, massage, supplements. However, for the great majority of these therapies no evidencebased (clinical randomised trials) results are available. CAM is usually used in addition to, and not as a substitute for conventional therapies. The motivation of patients to try CAM is complex; the willingness to take control of their healthcare, the desire to try everything available, the mass-media pressure and the erroneous notion that CAM is without risks. In fact, none of these treatments is totally devoid of risks. While the use of complementary and alternative modalities for the treatment of RA continues to increase, rigorous clinical trials examining their efficacy are needed before definitive recommendations regarding the application of these modalities can be made. PMID- 16380749 TI - [The arthropathy of systemic sclerosis]. AB - Joint involvement occurs in thirds of SSc patients during the course of the disease, but may be the onset manifestation. Arthralgias, stiffness and tendon sheath involvement constitute the most common clinical findings affecting all joints, but predominantly the fingers, wrists and ankles. The most common radiographic abnormalities in SSc patients are subcutaneous calcinosis and digital tuft resorptions, which are frequently observed at the hands. Juxtaarticular demineralisation, joint space narrowing and erosions also occur and are diagnostic challenges with rheumatoid arthritis. Flexion deformities and tendon friction rubs are more common in dcSSc; arthritis/arthralgias and radiographic abnormalities similarly affect patients from each subset. A recent classification of radiological patterns (inflammatory, degenerative, periarticular fibrotic) pointed out a greater prevalence of the fibrotic pattern at the hands and degenerative pattern at the feet. PMID- 16380750 TI - [Assessment of circadian rhythm in pain and stiffness in rheumatic diseases according the EMA (Ecologic Momentary Assessment) method: patient compliance with an electronic diary]. AB - BACKGROUND: Many researchers have used paper diaries in an attempt to capture patient experience. However, patient non-compliance with written diary protocols is a serious problem for researchers. Electronic patient experience diaries (eDiary) facilitate Ecologic Momentary Assessment (EMA) study designs by allowing the researcher to administer flexible, programmable assessments and mark each record with a time and date stamp. OBJECTIVES: The objectives of the current study were to evaluate methodological issues associated with real-time pain reports (EMA) using electronic patient experience diaries, to quantify compliance (percentage of the total number of diary reports scheduled that were actually completed), and to examine the circadian rhythm in pain and stiffness of patients with rheumatic diseases in an ecologically valid manner. METHODS: In this cross sectional study we examined 49 patients with rheumatic diseases (14 patients with rheumatoid arthritis, 18 with fibromyalgia and 17 with osteoarthritis of the knee), attending the care facilities of the Department of Rheumatology of Universita Politecnica delle Marche. All patients fulfilling the American College of Rheumatology (ACR) criteria. The assessment of pain and stiffness in all patients were repeated seven times a day (8 A.M., 10 A.M., 12 A.M., 2 P.M., 4 P.M., 6 P.M. and 8 P.M.) on seven consecutive days using an electronic diary (DataLogger(r) - Pain Level Recorder). A datalogger is newly developed electronic instrument that records measurements of pain and stiffness over time. Dataloggers are small, battery-powered devices that are equipped with a microprocessor. Specific software is then used to select logging parameters (sampling intervals, start time, etc.) and view/analyse the collected data. Compliance is based on the time and date record that was automatically recorded by the devices. RESULTS: Using the data from the electronic diary, we determined that the average verified compliance rate for pain and stiffness were 93.8 and 93.6%, respectively. The two highest compliance rates were observed in patients with rheumatoid arthritis (95.6 and 95.2%, respectively). There were no statistically significant difference in compliance between females and males or patients above or below 60 years old. Significant circadian rhythms in patients with RA and OA of the knee were detected in pain and stiffness. No rhythm in pain or stiffness was observed in subjects with fibromyalgia. CONCLUSIONS: We conclude that collection of subjective data using electronic diary in rheumatologic setting is a feasible method than can be adopted with high compliance rates across a range of patient demographic subgroups. The identification of diurnal cycles of self-reported pain and stiffness, using EMA method, has important implications for patients with respect to planning their daily activities and in developing individual therapeutic programs with respect to diurnal variability, which therefore may be more effective. PMID- 16380751 TI - [Soluble P-selectin levels in synovial fluid and serum from patients with psoriatic arthritis]. AB - OBJECTIVE: P-selectin is an adhesion molecule expressed by activated endothelial cells and platelets favouring the leukocyte adherence to microvascular endothelium. A soluble form of this molecule has been described, whose serum levels were found to be elevated and correlate with disease activity in rheumatoid arthritis (RA) patients. Aim of this study was to determine soluble P selectin levels in synovial fluid (SF) and serum from patients with psoriatic arthritis (PsA), where it has never been investigated, to define its involvement in PsA synovial damage. METHODS: we analysed, by ELISA, soluble P-selectin serum and SF levels in 100 patients presenting a knee joint effusion: 38 of them presented PsA, 40 RA and 22 osteoarthritis (OA). We examined the main clinical and laboratory parameters of these patients. Soluble P-selectin serum levels were also detected in 15 healthy subjects. RESULTS: soluble P-selectin SF levels were significantly higher in PsA and RA patients respect to OA subjects. Soluble P selectin SF levels were lower than those found in serum and the SF/serum ratio was higher in PsA and RA patients respect to OA. Soluble P-selectin serum levels were not significantly different among patients and controls. No correlation was found between SF and serum levels of soluble P-selectin and the main clinical parameters. CONCLUSIONS: our study of soluble P-selectin in PsA reveals a prominent local role of this molecule, with no differences respect to RA. Histological findings may be of help in understanding the role of this adhesion molecule in PsA. PMID- 16380752 TI - [Primary Sjogren's syndrome: clinical and serological feature of a single centre]. AB - OBJECTIVE: To describe clinical and serological features of a large series of patients affected by primary Sjogren's syndrome (pSS), assessing the evolution of the disease in a long-term follow-up study. METHODS: Clinical and laboratory data of 250 patients with pSS attending our Unit for a mean follow-up period of 140 months were retrospectively collected and analysed. In all the cases the diagnosis was made according with the recent international criteria. RESULTS: Glandular involvement was almost universally present, typically as the first manifestation of the disease and a slow progression of the salivary and lachrymal dysfunction was seen during the observation period. Extraglandular involvement was mild, quite rare and delayed. The respective percentages for muscle-skeletal disease, urogenital, haematological, skin, pulmonary, gastrointestinal, neurological and renal involvement were 60%, 40%, 24%, 20%, 11%, 7%, 8% and 3%. Only 6 patients developed a lymphoma. The serological pattern of the majority of patients remained constant throughout the follow-up period. CONCLUSIONS: pSS is often a benign condition. Since some patients may develop lymphoid malignancies, clinical follow-up is recommended. PMID- 16380753 TI - [Sensibility and specificity for pregnancy morbidity of anti-beta2-glycoprotein I antibodies in antiphospholipid syndrome]. AB - OBJECTIVE: This study aimed to evaluate the sensitivity and specificity of the anti-β2-glycoprotein I (GPI) antibodies for pregnancy morbidity in the antiphosoplipid syndrome (APS). METHODS: 335 women were recruited and on the basis of their clinical features were subdivided into 2 groups homogenous for number and age. The first (study) group contained the women whose pregnancy complications satisfied the classification criteria for APS. The second (control) group was made up of women with pregnancy complications not included in the classification criteria for APS. Anti-β2-GPI, anticardiolipin antibodies (aCL) and lupus anticoagulants (LA) were determined in all of these women. RESULTS: The only antiphospholipid antibodies occurring with a significant frequency (p=0,00) in the women with pregnancy criteria for APS were the IgG anti β2-GPI and the IgG aCL present respectively in 23,92% and in 27,60% of the women. Its association was found to be significant (p=0,000). The distribution of the different levels of positivity of the IgG and IgM anti-β2-GPI in the patients of the study and control groups was not significantly different. The highest sensitivity for pregnancy complications was that of the IgG aCL and of the IgG anti-β2-GPI whose difference was not statistically significant. The comparison of the specificity of the IgG and IgM anti-β2-GPI with that of the IgG and IGM aCL was not statistically significant. CONCLUSIONS: The importance of determining the IgG anti-β2-GPI as part of routine laboratory testing of women with pregnancy complications typical of APS was confirmed. Together with IgG aCL these antibodies have proved to be the most sensitive and specific markers of pregnancy complications in APS. PMID- 16380755 TI - [Prevalence of rheumatoid arthritis in North Sardinia: the Tempio Pausania's study]. AB - OBJECTIVE: To estimate the prevalence of rheumatoid arthritis in an Italian general population. METHODS: The study was conducted in the years 2002-2003 in Tempio Pausania's health district located on the North Sardinia and involved 30264 subjects aged 18 years or more treated by 29 general practices (GP). The GP was contact first by phone and letter and secondly a Rheumatologist administered a structured interview that included a screening questionnaire for RA. Cases were defined by the 1987 American College of rheumatology criteria adapted to epidemiological surveys. Every cases were examined by Rheumatologist and General practices together. The age and sex distribution of the sample were similar to those of the Italian population from the 2001 census. RESULT: Of the 141 subjects who fulfilled the inclusion criteria, there were 113 women and 28 men, with a mean age at diagnosis of 45 years. The prevalence of RA was 0,46% in the general population, 0,73% in women and 0,19% in men. CONCLUSION: Our results suggest that the prevalence of RA is comparable to that reported in other Mediterranean countries. Women are more affected than men. Our prevalence is higher than that observed in Chiavari in 1992. PMID- 16380754 TI - [Anti-TNFalpha treatment in patients with rheumatoid arthritis and anti-Ro/SSA antibodies]. AB - OBJECTIVE: To analyse clinical efficacy, onset of new autoantibodies or symptoms of autoimmune disease in patients affected by rheumatoid arthritis with anti Ro/SSA treated with anti-TNFalpha agents. METHODS: Six anti-Ro/SSA positive subjects with RA were studied every six months until 24th month of treatment in order to detect ANA titer (IFI), anti-dsDNA (Farr), anti-cardiolipin and anti beta2glycoprotein I (ELISA), anti-ENA (CIE). The titre of anti-Ro/SSA were analysed by ELISA. Four patients were diagnosed as overlap RA/SS. RESULTS: Six female patients (mean age 58 ys, SD 9.8 ys), with long-standing RA (mean 7 ys, range 5-22 ys) were treated with anti-TNFalpha agents for a mean of 31 months (SD: 20.4 m): 4 with Infliximab and 2 with Etanercept. All the patients showed a significant reduction of DAS until 24th month (p < 0.006) with stability of sicca symptoms. The titer of ANA and anti-Ro/SSA was stable, while 4 subjects developed anti-dsDNA at low titer within 6-12 months. One patient withdrawn the treatment, because of lupus-like features; another one, with HCV hepatitis, interrupted Etanercept because of elevation of liver enzymes. No anticardiolipin or antibeta2GPI antibodies were detected. One subject with RA-SS also presented a primary biliary cirrhosis: clinical and histological features of cholangitis remained stable during Etanercept treatment. CONCLUSIONS: Anti-TNFalpha treatment showed good efficacy and safety in anti-Ro/SSA positive patients with RA. Anti-ds DNA antibodies at low titer appeared in most patients while the onset of lupus like disease could be considered a rare event also in RA patients with a rich autoimmune repertoire. PMID- 16380756 TI - [The treatment of diffuse cutaneous systemic sclerosis with autologous hemopoietic stem cells transplantation (HSCT): our experience on 2 cases]. AB - OBJECTIVES: Autologous hematopoietic stem cell transplantation (HSCT) is a treatment option which may be considered for severe diffuse cutaneous systemic sclerosis (dcSSc) patients not responding to cyclophophamide (CY). We present two cases of dcSSc not responding to CY >10 g who were successfully treated with HSCT. PATIENTS AND METHODS: Two dcSSc patients were unresponsive to monthly i.v. pulse of CYC (0.75 g m2). Both patients had significant reduction of DLCO and mild-moderate pulmonary hypertension and HSCT was considered due to the rapid progression of the disease. Following informed consent and ethics committee approval, HSCT was performed. Mobilisation was performed with CY 4 g/m2 and recombinant human granulocyte colony stimulating factor (rHu GCSF) followed by a successful apheresis (CD34+ cells, >7X106). Conditioning regimens were: CY 100mg/kg body weight plus thiotepa 10 mg/ kg in the first patient and CY 200 mg/kg in the second. Both graft products were CD34 selected. No arrhythmias occurred during the procedure and no other severe side effects were observed during hospitalisation. RESULTS: Follow up: Patients underwent a monthly follow up with physical examination, pulmonary function tests and echocardiography every 3 months. Chest CT has been performed 6 months post transplantation. The following was observed: skin score (from 40 to 10 for the first patient and from 38 to 12 for the second one), LVEF and pulmonary function remained stable, PAP decreased from 45 mmHg to 35 mmHg and from 40 to 32 mmHg. No late complications or cardiac toxicity was observed. CONCLUSION: These two dcSSc cases demonstrate that HSCT may be successfully performed without serious side effects in cases in whom despite a cumulative CY dose was ineffective. This suggests an "immunological threshold" effect which may be exploited in other severe, therapy refractory autoimmune cases. PMID- 16380757 TI - [Psoriatic arthritis: epidemiological and clinical aspects in a cohort of 1.306 Italian patients]. AB - Because there is the impression that psoriatic arthritis is a composite disorder with mild forms close to more severe and aggressive ones, we conducted a multicenter study with the aim of characterizing disease expression in a large cohort of Italian patients. One-thousand-three-hundred-six patients fulfilled inclusion criteria and were analyzed in this study. Psoriasis antedated the onset of arthritis in the majority of the cases (67.7%). More rare was inverse or simultaneous onset which occurred in 17.3% and 15.0% of the cases, respectively. Peripheral articular involvement (mono-oligo or polyarthritis) was recorded in 88.7% of the cases while spondylitis occurred in 11.3%. Peripheral enthesopathies were found in 28.1% of the cases with a marked occurrence in patients with axial involvement (64.5% vs 35.5% in oligo or polyarthritis). Abnormal levels of ESR and CRP respectively occurred in 52.2% and in 52.6% of the cases, while rheumatoid factor was detected in 5.0% of the cases. On the basis of distribution of joint involvement, symmetry and presence of peripheral enthesopathies we recognized three clusters of arthritis. Patients included in Cluster 1 and Cluster 2 showed a severe form of polyarthritis in most of the cases (82.9%), with increased serum levels of inflammatory indices in more than 85% of the cases. Almost all the hospitalized patients (97.1%) were included in this two clusters. They markedly assumed steroids and methotrexate or another DMARD. About half of the patients (51.1%) included in Cluster 3 showed mono-oligo articular involvement. Serum inflammatory indices were increased in 20.8% of the cases while hospitalization occurred only in 2.9% of the cases and NSAIDs were the treatment of choice. The evidence in our country of a large prevalence of severe forms of arthritis needing specific and aggressive approach outlines the requirement of an intense educational action aimed at increasing the awareness of this condition. PMID- 16380758 TI - [The strange case of a patient affected by acromegaly with osteoporomalacia without hypogonadism]. AB - Acromegaly is a rare disease that, in the majority of cases, is due to the presence of a benign growth hormone (GH)-producing tumor of the pituitary. Growth hormone has profound effects on linear bone growth, bone metabolism, and bone mass. In acromegaly, the skeletal effects of chronic GH excess have been mainly addressed by evaluating bone mineral density (BMD). Most data were obtained in patients with active acromegaly, and apparently high or normal BMD was observed in the absence of hypogonadism. The Autors describe a case of patient affected by acromegaly without hypogonadism with serious osteoporosis and biological signs of osteomalacia. PMID- 16380759 TI - [Oral capillaroscopy: a new diagnostic method]. AB - The morphological-functional study of microcirculation is of fundamental importance; in fact, the microvascular bed is directly involved both in autoimmune etiopathogenesis pathologies, and in acute and chronic inflammatory etiopathogenesis pathologies. Oral capillaroscopy is a very stimulating method for studying microcirculation, because of the possibility of studying small vessels in vivo by means of a microscope. Today, it is becoming more reliable thanks to the improvement of the observation tools (photography, videomicroscopy). PMID- 16380760 TI - [Takayasu's arteritis. A concise review and some observations on a putative case reported by Giovanni Battista Morgagni (1761)]. AB - The discovery of Takayasu's arteritis is likely to date back as far as 1830, owing to the first description of the Japanese Rokushu Yamamoto. Thereafter, several authors from certain geographical areas and in various historical periods described such a vascular disorder, by introducing a quantity of definitions. At present, it is defined as an eponymic disease, namely Takayasu's arteritis, since Makito Takayasu, a Japanese ophtalmologist, reported in 1908 the clinical history of a woman showing some particular retinal anastomotic shunts of arterioles and venules. In the present study the description of an about 40 year-old woman suffering from a pulseless disease, as reported by Giovanni Battista Morgagni in 1761, is summarized. Such a description could be the first case report of Takayasu's arteritis, according to some previous literature data and our critical analysis. PMID- 16380761 TI - Is this clinical trial fully registered? A statement from the International Committee of Medical Journal Editors. PMID- 16380762 TI - The Ottawa Statement. International registration of protocol information and results from human trials of health related interventions. PMID- 16380763 TI - Ethical guidelines in genetics and genomics. An Islamic perspective. AB - We are at a time of unprecedented increase in knowledge of rapidly changing technology. Such biotechnology especially when it involves human subjects raises complex ethical, legal, social and religious issues. A World Health Organization expert consultation concluded that "genetics advances will only be acceptable if their application is carried out ethically, with due regard to autonomy, justice, education and the beliefs and resources of each nation and community." Public health authorities are increasingly concerned by the high rate of births with genetic disorders especially in developing countries where Muslims are a majority. Therefore, it is imperative to scrutinize the available methods of prevention and management of genetic disorders. A minimum level of cultural awareness is a necessary prerequisite for the delivery of care that is culturally sensitive, especially in Islamic countries. Islam presents a complete moral, ethical, and medical framework, it is a religion which encompasses the secular with the spiritual, the mundane with the celestial and hence forms the basis of the ethical, moral and even juridical attitudes and laws towards any problem or situation. Islamic teachings carry a great deal of instructions for health promotion and disease prevention including hereditary and genetic disorders, therefore, we will discuss how these teachings play an important role in the diagnostic, management and preventive measures including: genomic research; population genetic screening pre-marital screening, pre-implantation genetic diagnosis; assisted reproduction technology; stem cell therapy; genetic counseling and others. PMID- 16380764 TI - Assisted reproductive technology. What is needed in the Kingdom of Saudi Arabia? AB - In 1978, the first in vitro fertilization baby was born. Since then, we have seen the introduction of several new modalities of assisted reproductive technologies ART, and the spread of ART procedures throughout the world. The number of ART units in the Kingdom of Saudi Arabia is increasing rapidly. The associated complications of ART procedures include fetal, maternal and childhood. PMID- 16380765 TI - Serological diagnosis of autoimmune hepatobiliary diseases. AB - Autoimmune hepatobiliary diseases AIHBD comprise autoimmune hepatitis, primary biliary cirrhosis, primary sclerosing cholangitis and the overlap syndromes. Early diagnosis and treatment of AIHBD are essential for the prevention of the high morbidity, and mortality, that is, otherwise, associated with untreated patients. Screening for AIHBD relies heavily on the use of serological tests for the detection of serum autoantibodies that associate with the diseases. Understanding these tests, and the results produced, is important for the efficient diagnosis/exclusion of these diseases. In this review, we discuss the various tests available in the clinical immunology laboratory for screening of AIHBD and comment on the significance of the results produced by each test. We hope that this review will highlight this group of autoimmune diseases and lead to more efficient and earlier diagnosis, and treatment, of patients with AIHBD. PMID- 16380766 TI - Trace elements and oxidative stress in chronic obstructive pulmonary disease. AB - OBJECTIVES: Many trace elements have activator or inhibitor roles in the antioxidative defense systems in diseases such as chronic obstructive pulmonary diseases (COPD). In this study, we aimed to show the levels of trace elements with action in oxidative stress, and to show the levels of malondialdehyde (MDA) as a marker of oxidative stress, and the activity of paraoxonase (PON1) as an antioxidant in COPD and smokers. METHODS: We included 25 patients with COPD, and 20 healthy non-smokers in the study. We selected them from the hospitalized patients at the Hospital of Dicle University, Turkey, between April 2003 and January 2004. The clinical condition of the patients was stable. RESULTS: The serum copper (Cu) and MDA concentrations in COPD patients were higher than the control group. There were no differences in zinc (Zn) concentration and Cu/Zn ratio between COPD patients and the control group. We found lower serum PON1 activities in COPD patients compared with the control group. There was no statistically significant difference in Zn concentration and Cu/Zn ratio between smokers and non-smokers in COPD. There were statistically significant differences in Cu, MDA concentrations and serum PON1 activities between smokers and non smokers in COPD. CONCLUSION: We could suggest that trace elements such as Cu, oxidants and antioxidants such as MDA and PON1 have roles in oxidative stress, and in COPD. PMID- 16380767 TI - Frequency of methylenetetrahydrofolate reductase C677T polymorphism in patients with cardiovascular disease in Eastern Saudi Arabia. AB - OBJECTIVE: Homozygosity for the C677T mutation in the gene of the thermolabile enzyme 5,10 methylenetetrahydrofolate reductase (MTHFR) associates with reduced enzyme activity, leading to mild hyperhomocysteinemia. We now know that an elevated level of homocysteine is an important risk factor for cardiovascular disease (CVD). The objective of this study was to determine the prevalence of the C677T mutation in Saudi patients diagnosed with CVD. METHODS: Over a period of 2 years (2003-2004) in a case control study, we determined the prevalence of the C677T mutation in 83 CVD patients and in 40 age and gender-matched controls in the Eastern Province of Saudi Arabia. We determined the MTHFR genotype by restriction fragment length polymorphism and allele specific hybridization procedures. RESULTS: The CVD group showed over representation of the C677T allele frequencies (20.5%) compared with unaffected controls (15%) (p=0.3). Furthermore, the genotypic data indicated that the prevalence of homozygosity for the C677T mutation was dramatically higher in the CVD patients (10.8%) when compared with normal (0%) (p=0.058). CONCLUSION: These results suggest that the MTHFR C677T variant mildly influences CVD. However, we require further investigation in large independent samples. PMID- 16380768 TI - Breast carcinogenesis. Transition from hyperplasia to invasive lesions. AB - OBJECTIVES: To examine the balance loss between proliferation and apoptosis that play a role in breast cancer development, and to explore the places of various genes and molecules within this process in this supposed multistep process. METHODS: We obtained the specimens from 40 patients between 2002 and 2004 at the Department of Pathology, Medical Faculty, Adnan Menderes University, Aydin, Turkey. We categorized the lesions ductal hyperplasia (DH), atypical ductal hyperplasia (ADH), in situ ductal carcinoma (DCIS), and invasive ductal carcinoma (IDC). We determined the tumor size, histological grade and lymph node status of invasive cases and we used nottingham prognostic index (NPI). We applied ER, PR, c-erbB2, p53, Ki-67, bcl-2, dUTP nick end labeling (TUNEL), breast cancer gene-1, matrix metalloproteinases-1 and tissue inhibitor matrix metalloproteinases-1 stains to each lesion using the immunohistochemical method. RESULTS: We observed that ER and PR decreased in ADH when compared with DH (p=0.0001 and p=0.019). However, we determined that in DCIS as c-erbB2 (p=0.005) and Ki-67 (p=0.004) increase, TUNEL (p=0.04) and bcl-2 (p=0.005) decrease, when compared with ADH. When compared with DCIS lesions, we observed the existence of a higher c-erbB2 (p=0.003) and a lower TUNEL (p=0.012) in invasive tumors. Furthermore, we found that there is a higher MMP-1 (p=0.04) in invasive lesions, when compared with non invasive lesions. We detected higher PR (p=0.049), lower TUNEL and c-erbB2 (p=0.017) in low grade group of NPI, when compared with high grade group of NPI. CONCLUSION: As a result, it has been shown that together with increase in proliferation, decrease in apoptosis, too, contributes to the proliferation/apoptosis imbalance that occurs in breast carcinogenesis. Increase in proliferation and decrease in apoptosis are parallel with the progression of lesions. We also showed that the changes, beginning with loss of ER and PR in ADH step, can cause malign transformation, which is especially notable both in DCIS step due to Ki-67 and c-erbB2 increase, and also with bcl-2 and TUNEL decrease. PMID- 16380769 TI - Influence of ovariectomy and flutamide treatment on vaginal tissue and body weight. AB - OBJECTIVE: To determine the effect of flutamide on vaginal histology, and to ascertain whether flutamide regulates body weight gain in the ovariectomized rat model. METHODS: We chose 36 sexually mature female Wistar-Albino rats in this study. The study took placed in the Department of Medical Science Application and Research Center of Dicle University, Diyarbakir, Turkey, in 2002. We divided the model rats into 3 groups: group 1--control group, group 2--sham-operated group and group 3--bilaterally ovariectomized group. In addition, we gave flutamide to group 3. RESULTS: We found that the body weight was higher in the ovariectomized rats than the control rats. Furthermore, the body weight decreased a little after flutamide therapy. The thickness in the vaginal epithelium of the control group decreased, moreover, the appearance of stromal tissue was rather loose. After flutamide applications, infiltration of the stromal cells increased, the vaginal epithelial layer thickened and became keratinized. Microscopic papillae and anatomical processes appeared in the flutamide treated group. CONCLUSION: We observed that the antiandrogen drug flutamide is hormonally active and may exhibit estrogenic-antiestrogenic activity in rats. Our study may be a source for further research examining the relationship between the androgen receptor antagonist flutamide and reproductive function in female rats. PMID- 16380770 TI - Nonalcoholic fatty liver disease. Correlation with histology and viral hepatitis. AB - OBJECTIVE: Nonalcoholic fatty liver disease (NAFLD) involves a heterogeneous group of diseases that places some patients at risk of progression to cirrhosis. In this study, our aim was to investigate the relationships between the histopathological features of NAFLD, hepatic stellate cell activation, and capillarization to find a marker related to fibrosis, for NAFLD. METHODS: We studied liver biopsies from 62 patients with NAFLD, 21 patients with hepatitis B, and 19 patients with hepatitis C from the archives of the Department of Pathology, Gazi University Medical School, between 1997 and 2004. We performed immunoperoxidase stains for alpha-smooth muscle actin (alpha-SMA) and CD31 to identify activated hepatic stellate cells and capillarization. We investigated the relationships between histopathological features and both alpha-SMA and CD31 expressions. RESULTS: Most NAFLD cases were in low grades and stages. We found a relationship between both necroinflammatory grade and ballooning degeneration, and fibrosis. Pure steatosis did not relate to fibrosis. Immunohistochemical analysis revealed that CD31 expression was significantly higher than alpha-SMA expression in all groups. We determined a correlation between the fibrotic stage and CD31 expression, but not with alpha-SMA expression. In NAFLD cases, we detected the highest staining scores of CD31 in zone 3, while the portal/septal area was the dominant zone for control groups. There was no significant zone for alpha-SMA expression. CONCLUSION: Our results suggest that we can use CD31, rather than alpha-SMA, as a marker of endothelial damage and sinusoidal capillary transformation, both of which might precede fibrogenesis in chronic liver diseases, particularly in NAFLD. PMID- 16380771 TI - A new technique to avoid losing the strains of Echinococcus multilocularis during passaging. AB - OBJECTIVE: To examine the survival of Echinococcus multilocularis (E. multilocularis) and the formation of protoscoleces, under refrigerated conditions over a 43-day observation period. METHODS: We conducted this study in Ege University, School of Medicine, from May-December, 2004. We included 4 healthy females and 2 males, 8-12 week old M. unguiculatus species (approximately 60-70 gr) for the study, as they are known to be susceptible to E. multilocularis. In this experimental study, we aimed to define a technique to keep the strains during these passages. RESULTS: We found preserved viability and virulence of E. multilocularis stored in RPMI 1640 plus 10% fetal calf serum at +4 degrees C. We accomplished the infection of the Meriones unguiculatus after in-vivo passaging from that flask, which we kept at +4 degrees C, and this proves the viability of protoscoleces and membranous structures perfectly at the 43rd day. CONCLUSION: We also found that E. multilocularis metacestodes and the formation of protoscoleces keep their viability up to 60 days and virulence up to 43 days under this condition. We recommend storing an extra flask during each passage as a precaution against losing strains. PMID- 16380772 TI - Induction of clindamycin resistance in erythromycin-resistant, clindamycin susceptible and methicillin-resistant clinical Staphylococcal isolates. AB - OBJECTIVES: To demonstrate the in vitro ability of erythromycin to induce clindamycin in erythromycin resistant and clindamycin susceptible clinical isolates of Staphylococci. METHODS: We studied 291 clinical isolates of erythromycin-resistant (ER-R) clindamycin-susceptible Staphylococci (CL-S) at Almana General Hospitals, Al-Khobar, Dammam, Saudi Arabia during the period from June 2004 to May 2005. The isolates included 70 Staphylococcus aureus, 81 Methicillin Resistant Staphylococcus aureus (MRSA) and 140 coagulase-negative Staphylococci (CNS). We examined these isolates for inducible clindamycin resistance (ICR) by erythromycin induction test using double disc susceptibility test (D-test). Strains producing ICR show flattening of the clindamycin disc zone adjacent to the erythromycin disc. RESULTS: Of the 291 ER Staphylococci studied, 82 (28%) demonstrated constitutive clindamycin resistance [2 (2.9%) S. aureus, 43 (53%) MRSA and 37 (26%) CNS]. Inducible clindamycin resistance was demonstrated in 113 (38.8%) of Staphylococcal isolates, including 84 (28.9%) from adult patients and 29 (10%) from pediatric patients. The incidence of ICR was 49 (70%) for S. aureus, 35 (43%) for MRSA and 29 (20.7)% for CNS. Overall, 96 (33%) of the isolates remained susceptible to clindamycin and were negative for clindamycin induction [19 (27%) S. aureus, 3 (3.7%) MRSA and 74 (52.8%) CNS]. CONCLUSION: We conclude that a significant number of ER-R CL-S staphylococcal isolates studied were positive for ICR. These isolates should be reported as clindamycin resistant. Given the high rate of inducible resistance to clindamycin in the staphylococcal isolates, we recommend that microbiology laboratories perform erythromycin induction test on all ER-R CL-S staphylococcal isolates prior to reporting clindamycin susceptibility. PMID- 16380773 TI - Metabolic syndrome in Saudi Arabia. AB - OBJECTIVES: Metabolic syndrome (MS) is a well-established risk factor for the development of coronary artery disease (CAD). We designed this study to obtain the prevalence of MS and each of its components in Saudi Arabia. This study is part of Coronary Artery Disease in Saudi Study (CADISS). METHODS: We conducted this community-based national epidemiological health survey by examining Saudi subjects in the age group of 30-70 years of selected households over a 5-year period between 1995 and 2000 in Saudi Arabia. We interviewed all subjects, examined and took measurements of their blood pressure, weight, height, waist circumference, as well as fasting samples of plasma glucose, triglycerides, and high-density lipoprotein (HDL) cholesterol. We obtained the prevalence of MS based on the presence of at least 3 of the following: abdominal obesity (waist circumference > 102 cm (40 inch) in male and > 88 cm (35 inch) in female), triglycerides > or = 150 mg/dl (1.69 mmol/L), HDL cholesterol < 40 mg/dl (1.03 mmol/L) in male and < 50 mg/dl (1.29 mmol/L) in female, blood pressure > or = 130/85 mm Hg, fasting glucose > or = 110 mg/dl (6.1 mmol/L) as defined by the Adult Treatment Panel (ATP) III in 2001. RESULTS: We included 17,293 subjects in this survey during the study period. The overall age-adjusted prevalence of MS in Saudi Arabia obtained from this study is 39.3%. Age adjusted prevalence in males is 37.2% and crude prevalence is 40.9% (95% confidence interval [CI] 39.8-42), while females have a higher prevalence of 42% and crude prevalence of 41.9% (95% CI 40.9-42.9). Saudi subjects from urban areas have significantly higher prevalence of 44.1% (95% CI 43.2-45) compared to those living in rural areas of 35.6% (95% CI 34.3-36.7) (p<0.0001). Low HDL affects 81.8% of females and 74.8% of males with MS leading all other factors, and it continued to be consistent in all different age groups. Metabolic syndrome is a risk factor for CAD, as the prevalence of CAD was higher among patients with MS (6.7%) compared to subjects without MS (4.6%) (p<0.0001). CONCLUSION: The prevalence of MS is high in Saudi Arabia. Low HDL cholesterol plays a major role in the contribution to the MS in Saudi Arabia. Therefore, we recommend routine assessment for the components of MS in patients with CAD, furthermore, we encourage aggressive management of the MS for primary prevention of CAD, particularly, measures to increase HDL cholesterol. PMID- 16380774 TI - Chlamydia pneumoniae seropositivity and risk of developing coronary heart disease in Western Saudi Arabia. AB - OBJECTIVES: To estimate the seroprevalence of IgG and IgA antibodies against Chlamydia pneumoniae (C. pneumoniae) among a sample of the Saudi population, and to evaluate whether there is a relationship between seropositivity to chronic infection with C. pneumoniae and the manifestation of symptomatic coronary heart disease (CHD). METHODS: We collected 273 sera samples from CHD patients and 273 sera samples from healthy matched controls from the Western region of Saudi Arabia during the period from November 2004 to May 2005. We tested anti chlamydial IgG and IgA antibodies using enzyme-linked immunosorbent assay technique. RESULTS: We found 239 (87.5%) patients and 213 (78%) controls positive for C. pneumoniae IgG antibodies. However, 58 (21.2%) patients and 55 (23.9%) controls were positive for C. pneumoniae IgA antibodies. These results indicate a significant correlation between the presence of IgG antibodies and the development of CHD (p=0.003). Data of this study showed that the presence of IgG antibodies has a 2-fold increase risk in development of CHD. We found no significant correlation between the existence of IgA antibodies and CHD. CONCLUSION: Our study indicates that C. pneumoniae infection plays an important role in the development of CHD in the Saudi community, emphasizing the importance of developing strategies for prevention and control against this type of bacterial infection. However, we need further study throughout the Kingdom to approve these results in all regions. PMID- 16380775 TI - Effect of hormone replacement therapy on hemostatic variables in post-menopausal women. AB - OBJECTIVE: Hormone replacement therapy (HRT) has been increasingly promoted over the last 40 years to improve quality of life, and to reduce the risks of osteoporotic fractures and coronary heart disease (CHD). Recent randomized controlled clinical trials reported that HRT usage is associated with an increased risk of myocardial infarction (MI), stroke, and venous thrombosis. We conducted this study to evaluate the mean levels of some hemostatic parameters among groups that differ in estrogen levels and age. METHODS: We studied 150 healthy women in an observational comparative study, divided into 3 groups. Forty women were post-menopausal using HRT for a period of 6 months to 17 years. Fifty five women were post-menopausal and were not using HRT. Fifty-five women were younger pre-menopausal women with an age range of 20-54 years. The HRT group women were recruited from gynecologist private clinics while the other 2 groups were recruited in a random way from the society in Damascus, Syria between August 2002 and January 2003. We determined estradiol, fibrinogen, antithrombin III (AT III) and protein C in all women. RESULTS: When compared with post-menopausal non users group, current HRT users had higher mean levels of estradiol, but lower mean levels of AT III and protein C, and similar mean levels of fibrinogen. When compared with pre-menopausal group, current users had similar mean levels of estradiol, AT III and protein C, but higher mean levels of fibrinogen. However, post-menopausal non-users women had higher mean levels of fibrinogen and lower mean levels of AT III and protein C when compared with pre-menopausal women. CONCLUSION: Hormone replacement therapy treatment did not change fibrinogen mean levels, but it caused a decrease in AT III and protein C mean levels. PMID- 16380776 TI - Frequency and clinical characteristics of ketoacidosis at onset of childhood type 1 diabetes mellitus in Northwest Saudi Arabia. AB - OBJECTIVE: To determine the frequency, and to describe the clinical characteristics of ketoacidosis at initial diagnosis of childhood type 1 diabetes mellitus (T1DM) in Al-Madina region of the Northwest province of Saudi Arabia. METHODS: We retrospectively analyzed the hospital records of 311 (152 males and 159 females) children diagnosed with childhood T1DM in Al-Madina region, Saudi Arabia between January 1992 and December 2004. RESULTS: At diagnosis 172 (55.3%) children presented with diabetic ketoacidosis at the onset of their illness, 101 (58.7%) were females and 71 (41.3%) males. We found females to have more ketoacidosis at the onset of their illness with 1.4:1 female to male ratio. The mean age at presentation with ketoacidosis was 6.7 years (95% CI=5.6-7.8) ranging from 4 months to 14 years. Most of the ketoacidosis was mild to moderate (84.9%), while only 26 (15.1%) children had the severe type. Sixty-one (35.5%) children were in the younger age group, 54 (31.4%) were in the middle age group, and 57 (33.1%) were in older age group, there was no significant difference (p=0.5) between the 3 age groups in the frequency of ketoacidosis. The duration of symptoms before presentation with ketoacidosis was 15.8 days (95% CI=13.5-18.1). Altered consciousness was present in 21 (12.2%) children; all of them were from the severe type of ketoacidosis. There was a strong correlation between the severity of the central nervous system depression and the degree of acidosis (r=0.826, p<0.0001), but no correlation with age, gender, duration of symptoms, and blood glucose level. CONCLUSION: The frequency of ketoacidosis at onset of childhood diabetes mellitus in our region is significant. Prevention of diabetic ketoacidosis and reduction of its frequency should be a goal in managing children with diabetes. Rising standards of medical information and general awareness can contribute to this. PMID- 16380777 TI - Results and complications of external dacryocystorhinostomy surgery at a teaching hospital in Iran. AB - OBJECTIVES: To conduct a survey of outcomes, failure frequency rates, and other complications of dacryocystorhinostomy (DCR) at a teaching hospital in Iran. METHODS: In this outcome study, we survey the results of DCR in 187 consecutive chronic dacryocystitis patients operated at the Eye Department of Yazd Medical University, Iran from March 2001 to April 2003. We investigated the results and probable complications of this surgical procedure. RESULTS: Our data shows that the success rate of the operation was 90.4% with a confidence interval of 95% (86.3-94.5). The success rate was 91% in female and 89.3% in males (p=0.543). The failure rate was 9.6%. According to the methods used in this study, the success rate of the external method with silastic intubation was 96.3% and 88% without. These differences, however, were not statistically significant (p=0.08). The most frequent and important complication of DCR was failure and recurrence of epiphora or purulent discharge (9.6%). The less frequent complications included scar formation (8.6%), wound infection (5.3%) and granuloma formation (3.2%). There were no complications during the operation. CONCLUSION: External DCR is an easy technique with a success rate of more than 90%, with few complications. PMID- 16380778 TI - Validity of leukocyte count to predict the severity of acute appendicitis. AB - OBJECTIVE: To ascertain whether white blood cell (WBC) count with differential analysis may predict severity of disease in acute appendicitis. METHODS: We conducted this retrospective study on appendectomy patients from 1996 to 2001, at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. We reviewed patient's age, gender, duration of symptoms, temperature on admission, WBC count including differential and the histological diagnosis of the appendicular specimen. We further analyzed the data of those patients found to have acute, gangrenous and perforated appendicitis to determine the correlation between a high WBC count and a more advanced form of appendicitis. RESULTS: Out of an aggregate of 232 patients, 162 were males and 70 females with a mean age of 23.7 years (range, 12-70 years). Mean duration of symptoms was 1.9 +/- 1.1 days, mean temperature 37.8 +/- 1.4 degrees C, with reported elevated WBC count in 167 (71.9%) and normal in 65 (28.1%) cases. Mean WBC counts in acute were 14.5 +/- 7.3 x 10(9)/L, gangrenous 17.1 +/- 3.9 x 10(9)/L and perforated appendicitis 17.9 +/- 2.1 x 10(9)/L. This reflected a persistently higher WBC count in the complex (gangrenous, perforated) appendicitis compared with acute appendicitis (p < 0.05). The differential analysis showed neutrophilia in 123 (53%) and lymphopenia in 112 (48%) cases and out of these, 116 (94%) with neutrophilia and 107 (95%) with lymphopenia were reported to have appendicitis. CONCLUSION: A high WBC with differential count is a reliable indicator of the severity of appendicitis and signifies a more advanced stage. PMID- 16380779 TI - Drug poisoning in childhood. AB - OBJECTIVES: To determine and present the pattern of poisoning with drugs in Princess Rahmat Hospital, Irbid, Jordan, and to assess the effects of variables such as age, season, gender and agent on poisoning frequency. METHODS: We performed this retrospective cross-sectional study on 126 cases of poisoning with drugs hospitalized in Princess Rahmat Teaching Hospital, Northern Jordan, during a 6-year period from 1996 to 2001. The data collected included: age, gender, seasonal variation, material ingested, symptomatic or not on admission, time and place of ingestion, history of similar problem among the patient or his siblings, whether the parent induced vomiting before seeking medical help or not and the outcome regarding mortality and morbidity. We considered chi-square and values for the statistical analysis. RESULTS: There were 126 children (<12 years of age) admitted during the study period. Among the children, 54 (42.9%) were girls and 72 (57.1%) were boys. The majority (89.7%) was <6 years. There were seasonal variations of poisoning events with a higher frequency in the Spring (39%) and in Summer (35%). We considered severity mild in 60%, moderate in 27% and severe in 13% of cases. The large majority of drugs were psychotropic agents (56%) and principally diazepam's (39%). Self-poisonings were the most frequent cause of poisonings among children <12 years of age (mostly accidental poisonings such as drug poisonings). None of the children died or had a permanent sequel. CONCLUSION: Although we recorded no deaths in this study, drug poisoning in children remains a frequent problem, highlighting the need to develop an education program on primary prevention in our region. Parental education and intensified child supervision are the indicated measures of prevention for unintentional poisoning. PMID- 16380780 TI - Incidence of chromosome abnormalities in the Sultanate of Oman. AB - OBJECTIVES: To evaluate the cytogenetic findings in Omani children referred for suspected chromosomal anomalies that caused a variety of clinical disorders. Secondly, to study the frequency of chromosomal abnormalities in these patients and to compare our results with those reported elsewhere. METHODS: We performed chromosomal analysis on 1800 consecutive pediatric patients referred to the Cytogenetics section between June 1999 and May 2004 at Central Public Health Laboratories, Sultanate of Oman. Indication for referrals for exclusion of chromosomal rearrangements was multiple congenital anomalies, dysmorphic features, unclassified mental retardation, developmental delay, growth, and endocrine disorders. We carried out the lymphocyte culture according to standard methods. RESULTS: We found various types of chromosomal anomalies in 510 (28.3%) children and showed abnormal karyotypes in the form of trisomy 21 (391; 21.7%), trisomy 18 (32; 1.8%), trisomy 13 (20; 1.1%), sex chromosome aberrations (50; 2.8%) and other types of abnormalities (17; 0.95%). There was a considerable phenotypic-cytogenetic heterogeneity. We found a high rate of chromosomal abnormalities in the present study, and we observed variations in the frequency of chromosomal aberrations reported by different investigators. CONCLUSION: The higher incidence of the chromosomal abnormalities demonstrates the importance of cytogenetic evaluation in patients with dysmorphic features and congenital anomalies. Our findings suggest that chromosome analysis is a useful tool in the investigation of children with genetic disorders of unknown origin for confirmation of clinical diagnosis and proper medical care followed by genetic counseling and management. PMID- 16380781 TI - Prevalence, knowledge, beliefs and psychosocial impact of acne in University students in Central Saudi Arabia. AB - OBJECTIVES: To describe the prevalence of acne among Qassim University students, and to evaluate the knowledge, beliefs, and its psychosocial impact on them. METHODS: We observed 717 students (381 males, and 336 females), at Qassim University Medical clinics during February and March 2005. We interviewed and examined the subjects for the presence of acne. After confirming the diagnosis, we asked the patients several questions about acne, history, knowledge, perceptions, beliefs, and its psychosocial impact on them. We performed statistical analysis using the chi-square test with a 5% significance level. RESULTS: We found 56.2% of the students to have acne. The difference between both gender was statistically insignificant, and 47.9% of patients suffered from acne for more than one year. Of those who sought medical advice, 40.3% had their consultation within 3 months of the disease onset, and 58.9% of patients sought medical advice as a self-decision. Fifty-six percent believed they have an adequate knowledge of acne, and the most common source of information was newspapers. The most believed factor responsible for acne was hormones, and the most aggravating factor was stress. In 46% of patients, acne had no, or minimal effect on their self-image and in most of the patients (73%) it had no or minimal effect on their relationships. CONCLUSION: Acne is a common skin disease among Qassim university students, affecting both gender. We need health education in our community to encourage people to seek appropriate help for skin problems. We require further community based research to evaluate the effectiveness of such educational interventions in under-standing the natural history, pathogenesis and the sequelae of acne, increasing help-seeking behavior, and improving the awareness of patients about acne. PMID- 16380782 TI - Prevalence of visual deficits among young men in Jordan. AB - OBJECTIVE: To report the pattern of various causes of decreased visual acuity (VA), within the age group of young (18-25 years) healthy adults in Jordan. METHODS: We carried out this retrospective study on the records of 16,550 candidates examined from June 2004 to June 2005 by the Medical Committee of Employment, Royal Medical Services in Amman, Jordan. All candidates received an ophthalmic examination as well as medical and ENT examination. We divided the candidates with visual acuity of less than 6/12 in either eye into 3 groups. RESULTS: The most important cause of decreased VA in the 18-25 years age group was refractive errors followed by amblyopia, which represents the major ophthalmic cause of employment rejection in a wide range of occupations in adult life. CONCLUSION: This study emphasize the need for early visual screening for refractive errors and anisometropic amblyopia in early primary schools, and every effort must be made to achieve the best possible acuity in young patients with amblyopia. PMID- 16380783 TI - Prevalence of tongue lesions among Turkish schoolchildren. AB - OBJECTIVES: To collect data on the prevalence of tongue lesions of Turkish schoolchildren in 3 different population samples in Ankara, and to analyze the relationship between tongue lesions and gender, oral hygiene, and income levels. METHODS: Nine hundred and six schoolchildren aged 6-12 participated in this study between March 2004 and July 2004 in Ankara, Turkey. Of the 906 schoolchildren, 442 were girls and 464 were boys from 3 primary schools. We examined all the children for the presence of the following tongue lesions: 1) ankyloglossia, 2) bifid tongue, 3) fissured tongue, 4) geographic tongue, 5) median rhomboid glossitis, 6) lingual thyroid nodule, 7) atrophic tongue, 8) hairy tongue, 9) crenation tongue, 10) furred tongue, 11) macroglossia, 12) microglossia, and 13) lingual or sublingual varicosities. RESULTS: Of the 906 subjects, we detected 45 subjects with tongue lesions with a prevalence of 4.95%. We found lesions more frequently in boys (6%) than in girls (4.5%). However, these results were statistically insignificant. Oral lesions commonly found were geographic tongue (1.8%), ankyloglossia (1.3%), fissured tongue (0.9%), bifid tongue (0.4%), hairy tongue (0.2%) and macroglossia (0.2%), lingual thyroid nodule (0.1%). We did not find any of the following lesions: median rhomboid glossitis, atrophic tongue, crenation tongue, furred tongue, microglossia, lingual, or sublingual varicosities. CONCLUSION: The tongue has a special importance due to its localization. It can readily affect a patient's social life because of lesions and various symptoms. The localization of tongue may also play important role in diagnosis for systematic, hormonal, and allergic disorders. PMID- 16380784 TI - Measles seroepidemiology in 3 cities in Turkey. AB - OBJECTIVE: To evaluate immunity against measles and its relation with some variables among healthy subjects in 3 cities in Turkey. METHODS: We carried out a cross-sectional study on measles antibody titers in the serum samples of 712 people from Antalya, 696 from Diyarbakir and 667 from Samsun, Turkey using particle agglutination test between February 2000 and October 2001. The study groups, informed and asked for their consent by midwives, consisted of randomly selected subjects of all ages older than 6 months. We implemented the study in 3 steps: physical examination, interview and blood collection. RESULTS: We considered titers of > or = 1:16 as positive and we observed lower seropositivity in Diyarbakir (90.8%) than Antalya (95.9%), and Samsun (94.9%) (p=0.040). We also observed that seropositivity was lower among preschool group than older groups (p=0.006). The number of doses of measles vaccine (p=0.001) and measles infection history (p=0.003) were found as a factor increasing the seropositivity ratio. There was no statistically significant between age groups (p=0.219), gender (p=0.148), residence (p=0.537), and number per household (p=0.983) among the provinces. CONCLUSION: Based on the findings, measles infection still has a high incidence in Turkey and the second dose of measles vaccine is extremely important in the prevention of measles transmission among school children and the community. Furthermore, we must improve our regional differences in routine vaccination services. PMID- 16380785 TI - Ossifying renal tumor of infancy. AB - Ossifying renal tumor of infancy ORTI is a benign neoplasm, which presents with gross hematuria and less frequently as an abdominal mass, histologically it comprises a large cell with an epithelial nature and osteoid formation. We report a case of a 10-month-old girl who developed ORTI as non-opacified upper calyces of left kidney on excretory urography. We outlined the calcific and tumors nature of the lesion by ultrasound and computed tomography. The macroscopic and histological features were pathognomonic. PMID- 16380786 TI - Anesthesia management in Kabuki make-up syndrome. AB - Kabuki make-up syndrome KMS is a rare condition with a number of characteristic congenital abnormalities. The syndrome is characterized by peculiar facial appearance resembling the make-up of actors in Kabuki, the traditional Japanese theater, skeletal anomalies, dermatoglyphic abnormalities, postnatal growth deficiency, and mental retardation. These are rare reports of central nervous system dysfunctions, other than mental retardation, and no previously described congenital talipes calcaneo-valgus in this syndrome. We report the case of a 22 month-old girl having Kabuki make-up. At presentation, she had an adenoid hypertrophy and a history of recurrent otitis media. She had also delay in motor development, and a postnatal growth deficiency. The variable phenotypic expression is a well-known characteristic of the syndrome. For that reason, we should perform careful morphologic examination in every patient and their parents, and use flexotype laryngoscope Heine, Germany to visualize vocal cord in case of difficult intubation. At preoperative examination, as clinicians, we must be careful regarding patient morphology. Congenital heart defects and epilepsy are important for anesthesia management in KMS. PMID- 16380787 TI - Ascites and eosinophilic colitis in a young patient. AB - A 32-year-old man presented with 3-weeks history of abdominal pain and distention. Physical examination showed ascites, with no stigmata of chronic liver disease. Cytological preparations from the ascitic fluid showed a heavy population of mature eosinophils. Histological examination of colonic biopsies revealed a heavy expansion of the mucosa by sheaths of eosinophils. On the following days, the peripheral eosinophilia, ascites and abdominal pain resolved spontaneously. PMID- 16380788 TI - A rare presentation of hydatid cyst. AB - Hydatid disease, although known to occur in most body areas, is extremely rare in the female reproductive system. There are different modes of presentation for the disease; however, we report and discuss a case presented with cystic vesicles passing through the vagina, which is considered as a rare presentation for secondary involvement of the uterus and both ovaries. We confirmed diagnosis with radiological examinations and serological tests. We operated on the patient, and studied the excised cysts microscopically. The gynecologist should be aware of hydatid cyst when vaginally passing a grape like vesicle is presented by the patient. PMID- 16380789 TI - Successful treatment of chronic persistent vesicular hand dermatitis with topical pimecrolimus. AB - Hand dermatitis is a common chronic skin condition that has many clinical forms including contact, hyperkeratotic, frictional, nummular, atopic, pompholyx and chronic vesicular hand dermatitis. Topical steroids are the first line agents used. Here, we report the successful response to topical pimecrolimus 1% cream in a patient with steroid resistant chronic vesicular hand dermatitis. PMID- 16380790 TI - A death due to electrical injury through external ear canals. PMID- 16380791 TI - The incidence and impact of lupus anticoagulants among patients in the intensive care unit. PMID- 16380792 TI - Utility of cefoxitin resistance determination by disk diffusion method for routine detection of methicillin resistant Staphylococcus aureus. PMID- 16380793 TI - The hypo-hyper syndrome. PMID- 16380794 TI - Actinomyces meyeri isolation from synovial fluid of a patient with metastatic squamous cell lung carcinoma. PMID- 16380795 TI - Role of maternal factors in the etiology of neural tube defects in Jordan. PMID- 16380796 TI - Self-inflicted needle in the urethra retrieved endoscopically from the bladder. PMID- 16380797 TI - Torsion of an epiploic appendix mimicking acute appendicitis. PMID- 16380798 TI - Public knowledge and attitudes towards passive smoking. PMID- 16380799 TI - The evaluation of the internet and computer utilization by the fourth grade medical students. PMID- 16380800 TI - Evaluation of oxacillin and cefoxitin disk diffusion test for routine detection of methicillin resistant Staphylococcus aureus. PMID- 16380801 TI - Treatment of vitiligo with topical 15% lactic acid solution in combination with ultra violet-A. PMID- 16380803 TI - Paradoxical rise in serum adiponectin concentration in the face of acid-induced insulin resistance 13-cis-retinoic. AB - AIMS/HYPOTHESIS: We previously reported that treatment of acne with 13-cis retinoic acid causes insulin resistance and disturbances in lipid metabolism resembling those of the insulin-resistance syndrome. It is not known whether this is associated with alterations in the concentrations of serum adiponectin, an insulin-sensitising hormone secreted by adipocytes. MATERIALS AND METHODS: Eleven men (age 24+/-2 years, BMI 22.1+/-0.9 kg/m(2)) received 13-cis-retinoic acid (Roaccutan) treatment for acne for an average of 5 months. The insulin sensitivity of the subjects and concentrations of serum adiponectin were measured before, during and 1 month after the treatment by a euglycaemic-hyperinsulinaemic clamp and ELISA, respectively. RESULTS: There was a reversible reduction in whole body insulin sensitivity during therapy with 13-cis-retinoic acid. This was associated with a transient 34% increase in serum adiponectin concentration (from 5.3+/-0.9 to 7.1+/-1.2 mug/ml, p<0.05), with a return to pretreatment levels by 1 month after the end of therapy. In the pretreatment study, as well as in the study performed 1 month after the end of therapy, there was a small yet significant decrease in serum adiponectin concentration during a 4-h euglycaemic hyperinsulinaemic clamp. This decrease was not observed in the clamp performed during treatment with 13-cis-retinoic acid. CONCLUSIONS/INTERPRETATION: There is a paradoxical increase in fasting serum adiponectin concentration during the 13 cis-retinoic acid-induced reduction in insulin sensitivity. PMID- 16380804 TI - The metabolic syndrome, babies and bathwater. PMID- 16380802 TI - Changes in hepatic glycogen cycling during a glucose load in healthy humans. AB - AIMS/HYPOTHESIS: Glycogen cycling, i.e. simultaneous glycogen synthesis and glycogenolysis, affects estimates of glucose fluxes using tracer techniques and may contribute to hyperglycaemia in diabetic conditions. This study presents a new method for quantifying hepatic glycogen cycling in the fed state. Glycogen is synthesised from glucose by the direct and indirect (gluconeogenic) pathways. Since glycogen is also synthesised from glycogen, i.e. glycogen-->glucose 1 phosphate-->glycogen, that synthesised through the direct and indirect pathways does not account for 100% of glycogen synthesis. The percentage contribution of glycogen cycling to glycogen synthesis then equals the difference between the sum of the percentage contributions of the direct and indirect pathways and 100. MATERIALS AND METHODS: The indirect and direct pathways were measured independently in nine healthy volunteers who had fasted overnight. They ingested (2)H(2)O (5 ml/kg body water) and were infused with [5-(3)H]glucose and acetaminophen (paracetamol; 1 g) during hyperglycaemic clamps (7.8 mmol/l) lasting 8 h. The percentage contribution of the indirect pathway was calculated from the ratio of (2)H enrichments at carbon 5 to that at carbon 2, and the contribution of the direct pathway was determined from the (3)H-specific activity, relative to plasma glucose, of the urinary glucuronide excreted between 2 and 4, 4 and 6, and 6 and 8 h. RESULTS: Glucose infusion rates increased (p<0.01) to approximately 50 mumol kg(-1) min(-1). Plasma insulin and the insulin : glucagon ratio rose approximately 3.6- and approximately 8.3-fold (p<0.001), respectively. From the difference between 100% and the sum of the direct (2-4 h, 54+/-6%; 4-6 h, 59+/-5%; 6-8 h, 63+/-4%) and indirect (32+/-3, 38+/-4, 36+/-3%) pathways, glycogen cycling was seen to be decreased (p<0.05) from 14+/-4% (2-4 h) to 4+/-3% (4-6 h) and 1+/-3% (6-8 h). CONCLUSIONS/INTERPRETATION: This method allows measurement of hepatic glycogen cycling in the fed state and demonstrates that glycogen cycling occurs most in the early hours after glucose loading subsequent to a fast. PMID- 16380805 TI - Differential gene expression in a paclitaxel-resistant clone of a head and neck cancer cell line. AB - The anti-neoplastic drug paclitaxel (taxol), which is known to block cells in the G2/M phase of the cell cycle through stabilization of microtubules, is meanwhile commonly used for chemotherapy of advanced head and neck cancer. Chemotherapy is primarily used in order to preserve laryngeal and/or pharyngeal structures. Although paclitaxel generally seems to be a powerful agent, it failed to reach a loco-regional tumor control in a sufficient percentage of patients. In order to investigate molecular resistance mechanisms, we have established a paclitaxel resistant subline originating from the larynx carcinoma cell line HLaC79, which seemed to be partially dependent on taxol. The original and the descendant cell line were characterized by growth inhibition assays. We used western blotting and the cDNA subtraction (SSH) technique to identify genes differentially expressed in the taxol-resistant cell clone. cDNA subtraction revealed increased expression of six genes, including clathrin heavy chain, alpha3-tubulin, a neuroblastoma specific Thymosin beta, the ribosomal protein L7a, HLA-B associated transcript 3 and collagen IIIalpha1 in the taxol-resistant cell line. Furthermore, western blots showed an overexpression of MDR-1 in the taxol-resistant clone, while alpha and beta-tubulins and p48/IRF9 were expressed in equal amounts in both cell lines. PMID- 16380806 TI - Changes in distortion of two-tone cochlear microphonic and otoacoustic emission signals during an acute endolymphatic hydrops in the guinea pig. AB - An acute endolymphatic hydrops was induced by the injection of 1.1 mul of artificial endolymph into the scala media of guinea pig cochleas. This volume corresponds with an acute endolymphatic hydrops of 23%. During and after the injection, cochlear function was assessed by measuring the 2f(1)-f(2 )and f(2) f(1) distortion products in cochlear microphonics (CMDP) and the 2f(1)-f(2) distortion product otoacoustic emission (DPOAE). A reversible pressure increase of 23 Pa and a relatively stable endocochlear potential (EP) were accompanied by a mean decrease in 2f(1)-f(2) DPOAE of only 3.4 dB. Similarly, the 2f(1)-f(2) CMDP amplitude change was minimal during and after the injection. The only substantial change was measured in the f(2)-f(1) CMDP amplitude. The measured range of distortion amplitudes during an acute endolymphatic hydrops can be related to small changes in the cochlear transducer operating point. PMID- 16380807 TI - Neurological complications following functional neck dissection. AB - A retrospective study was done to assess the incidence and factors associated with neurological complications in patients who have undergone a functional neck dissection (FND). Four hundred forty-two epidermoid cancer patients operated on from January 1984 to December 2002 were included in the study. Clinical parameters, neurological sequelae, and other complications were evaluated in all cases. The incidence of neural damage was calculated on the nerves at risk (n =714). Paralysis of the 11th nerve occurred in 12 cases (1.68%). A lesion of the marginal branch of the 7th cranial nerve was observed in nine cases (1.26%). Bernard-Horner's syndrome and hypoglossal nerve paralysis were noted in four and three cases (0.56 and 0.42%), respectively. Thus, the incidence of neurological sequelae after FND is low. Neurological complications were not associated with either clinical parameters or non-neurological complications (P >0.05). None of the factors studied can predict the appearance of neural problems in the postoperative period. PMID- 16380808 TI - A new labyrinthulid isolate that produces only docosahexaenoic acid. AB - We show here that a new labyrinthulid strain, L72, isolated from a fallen leaf in the Seto Inland Sea of Japan, produced only docohexaenoic acid (DHA) among all the long-chain polyunsaturated fatty acids (LCPUFAs). The main fatty acid composition was 16:0 (28.9%), 18:0 (7.2%), 18:1 (5.7%), 18:2 (10.4%), and DHA (45.9%) without any other LCPUFA. The lipid content of the strain was 27.4%. The cells had many lipid bodies, which were densely located in all of the cells. On phylogenetic analysis using the 18S rDNA sequence, the strain was located in the labyrinthulids group, forming a monophyletic group with Labyrinthula sp. (strain s) and Labyrinthuila sp. (strain L59). We further tested the culture optimization of strain L72 to evaluate the ability of the strain to produce DHA. The optimum salt concentration and the temperature of the strain were 100% of artificial seawater and 20 degrees C. Strain L72 could grow well on soybean oil (SBO) or soybean lecithin (SBL) as the carbon source. When 20 g/l of SBL was added to the medium, DHA production reached the maximum amount at 0.67 g/l for 14 d. The two important facts, that the strain can use SBL as the main nutrient and contains only DHA among the LCPUFAs, will be of great advantage for industry. PMID- 16380809 TI - Molecular detection of marine invertebrate larvae. AB - The ecological patterns of many invertebrate larvae remain an ongoing mystery, in large part owing to the difficult task of detecting them in the water column. The development of nucleic-acid-based technology has the potential to resolve this issue by direct identification and monitoring of embryonic and larval forms in situ. We report herein on the successful development and application of nucleic acid-based sandwich hybridization assays that detect barnacles using rRNA targeted probes with both group-(order Thoracica) and species-(Balanus glandula) specificity. Primary results include the determination of target 18S rRNA sequences and the construction of "capture" probes for detection of larvae using hybridization techniques. In addition, we modified existing protocols for whole cell hybridization of invertebrate larvae as confirmation of the sandwich hybridization results. We used both hybridization techniques successfully in the laboratory on a plankton time series collected over 3 months, as well as a week long in situ deployment of the technique in Monterey Bay, CA. The adaptability of this technology promises to be further applicable to various organisms and could be used to enhance our understanding of larval presence in the world's oceans. PMID- 16380810 TI - ICTV in San Francisco: a report from the Plenary Session. PMID- 16380811 TI - Human cytomegalovirus UL131A, UL130 and UL128 genes are highly conserved among field isolates. AB - Coding sequences of the UL131A, UL130, and UL128 genes of human cytomegalovirus (HCMV) were found to be highly conserved among 34 field isolates from pregnant women with primary HCMV infection and their fetuses or newborns, as well as from solid organ transplant recipients and patients with AIDS. No strain clustering was observed. In contrast, sequencing of UL55 (gB coding gene) allowed the 34 isolates to be clustered into 4 genotypes. The conservation of the UL131A-UL128 locus is consistent with the conclusion that the three encoded proteins are all essential for growth of HCMV in endothelial cells and virus transfer to leukocytes. PMID- 16380812 TI - Characterization and partial genome sequence of stocky prune virus, a new member of the genus Cheravirus. AB - Characterization of a seemingly new spherical virus isolated from severely affected plum trees in south-western France indicated that its divided genome is composed of two single-stranded, polyadenylated RNAs of approximately 7.4 and 3.7 kb. Its particles are composed of three coat protein subunits of approximately 23, 23.5, and 24.5 kDa. Partial sequencing of the genomic RNAs indicated that this new virus, tentatively named stocky prune virus (StPV), is distantly related to the two sequenced cheraviruses, cherry rasp leaf virus (CRLV) and apple latent spherical virus (ALSV). StPV should be regarded as a new member in the unassigned genus Cheravirus. PMID- 16380813 TI - Antenatal psychiatric illness and adequacy of prenatal care in an ethnically diverse inner-city obstetric population. AB - OBJECTIVE: This study assessed the prevalence of antenatal psychiatric illness in low-income, ethnically diverse patients in an urban obstetric clinic and examined associations between positive psychiatric screens and inadequate utilization of prenatal care. METHODS: Bilingual research assistants administered the Primary Care Evaluation of Mental Disorders Patient Health Questionnaire and the Mood Disorder Questionnaire to 154 English- and Spanish-speaking pregnant patients attending routine prenatal visits. We assessed associations between patient characteristics, current and past psychiatric diagnoses, and utilization of prenatal care. RESULTS: Forty-five (29%) women screened positive for criteria for current psychiatric disorders with the highest rates for major or minor depression (26%) and anxiety disorders (10%). Inadequate prenatal care utilization was significantly associated with past psychiatric history and domestic abuse in the last year, but not with current psychiatric diagnosis, alcohol abuse, age, primiparity, marital status, receipt of government assistance, or unplanned pregnancy. Even after adjustment for possible confounding risk factors (e.g. past substance abuse, single marital status, unstable housing, education less than high school, and having other children), past psychiatric history was still significantly associated with inadequate prenatal care utilization and delayed initiation of care. CONCLUSIONS: A high percentage of disadvantaged pregnant women meet screening criteria for psychiatric disorders when screened during routine prenatal visits. Screening for past psychiatric history in routine prenatal visits could identify patients at risk for inadequate utilization of prenatal care. PMID- 16380814 TI - Innate sensors of microbial infection. AB - The innate immune system utilizes multiple families of pattern-recognition receptors (PRRs) to protect the host from infection. Each of these families contributes certain elements to the complement of innate effector functions that is elicited during an infection. Here we review the families of PRRs and explore examples of their cooperativity. PMID- 16380815 TI - Manifold mechanisms of Toll-like receptor-ligand recognition. AB - Toll-like receptors recognize a diverse range of molecules derived from pathogens as well as host cells. As the number and diversity of TLR ligands and host factors increase, more questions are being raised. Here, we review recent advances toward understanding the molecular and cellular mechanisms underlying TLR-mediated direct or indirect recognition of their diverse range of ligands, including lipids, proteins, and nucleic acids. The elucidation of such mechanisms may represent a key for developing novel immunotherapeutics for infectious diseases, allergies, or cancer and to intervene in immunological disorders such as autoimmune diseases. PMID- 16380816 TI - V alpha14 i NKT cells are innate lymphocytes that participate in the immune response to diverse microbes. AB - Natural Killer T (NKT) cells constitute a conserved T lymphocyte sublineage that has been implicated in the regulation of various immune responses, including the responses to viruses, bacteria, and parasites. NKT cells recognize self and foreign glycolipids presented by CD1d, a non-classical antigen-presenting molecule, and they rapidly produce various cytokines. Many studies have shown that NKT cells have protective roles following microbial infection through the amplification of innate and adaptive immunity, although NKT cells have detrimental roles in some cases. Recent studies have shed light on the natural antigens recognized by NKT cells and the mechanisms whereby they contribute to host defense, and they suggest that these unique T cells have evolved to jump start the immune response to microbes. PMID- 16380817 TI - NKG2D in NK and T cell-mediated immunity. AB - One of the best characterized NK cell receptors is NKG2D, a highly conserved C type lectin-like membrane glycoprotein expressed on essentially all NK cells, as well as on gammadelta-TcR+ T cells and alphabeta-TcR+ CD8+ T cells, in humans and mice. Here we review recent studies implicating NKG2D in T cell and NK cell mediated immunity to viruses and tumors, and its potential role in autoimmune diseases and allogeneic bone marrow transplantation. PMID- 16380818 TI - The anti-death machinery in IKK/NF-kappaB signaling. AB - The most extensively studied function of NF-kappaB is its ability to promote cell survival through induction of target genes, whose products inhibit various aspects of the apoptotic machinery in both normal and malignant cells. Recent studies, however, indicate that NF-kappaB activation can also suppress programmed necrosis through induction of genes encoding anti-oxidant proteins. Since tumor cells often use NF-kappaB pathway as a shield to escape the killing of conventional anti-cancer therapies, intervention of IKK/NF-kappaB signaling would be a promising option to improve the efficacy of cancer treatment. PMID- 16380820 TI - Probabilistic regulation of IL-4 production. AB - Among a population of uniformly differentiated TH(2) cells, only a portion express IL-4 upon stimulation and those that do often express the product of only a single allele. We review the evidence for the basis of IL-4 monoallelism and argue that it depends upon probabilistic expression of the Il4 gene. Further, we argue that probabilistic expression may provide a powerful mechanism through which certain key functions of IL-4, such as immunoglobulin class switching and determination of macrophage phenotype, may be efficiently regulated. PMID- 16380821 TI - Protective molecules--C-reactive protein (CRP), serum amyloid P (SAP), pentraxin3 (PTX3), mannose-binding lectin (MBL), and apolipoprotein A1 (Apo A1), and their autoantibodies: prevalence and clinical significance in autoimmunity. AB - Apoptotic defects and impaired clearance of cellular debris are considered key events in the development of autoimmunity, as they can contribute to autoantigen overload, and may initiate an autoimmune response. The pentraxins are a group of highly conserved proteins including the short pentraxins, C-reactive protein (CRP) and serum amyloid-P (SAP), and the long pentraxin-3 (PTX3), which are all involved in innate immunity and in acute-phase responses. Mannan-binding lectin (MBL) is an activator of the complement system, and Apolipoprotein A-1 (Apo A-1) is pivotal in the cholesterol homeostasis and has anti-inflammatory properties. In addition to their role in innate immunity and inflammation, each of these five proteins participates in the removal of damaged and apoptotic cells. In this review, we discuss the clinical significance of different levels of these proteins, their role in the induction or protection from autoimmunity, and the presence of specific autoantibodies against them in the different autoimmune diseases. PMID- 16380822 TI - Molecular aspects of regulation of collagen gene expression in fibrosis. AB - Fibrosis, the hyper-accumulation of scar tissue, is characterized by the overproduction and deposition of type I and III collagen by fibroblasts and is the one of the main pathologic outcomes of the autoimmune disorder scleroderma. While the causes of fibrosis in scleroderma are unknown, cytokines such as TGF beta, IL-4 and IL-13, play a crucial role in the stimulation of collagen production have been implicated in the disease process. In fibroblasts stimulation of collagen production by these cytokines is dependent on the Smad and STAT6 signaling pathways induced by TGF-beta and IL-4, IL-13 respectively. Furthermore, mounting evidence suggest cytokine crosstalk is relevant in the sclerotic process. Our laboratory demonstrated an increase in TGF-beta1 gene transcription from fibroblasts stimulated with IL-4. In addition, TSK/+ mice lacking the IL-4alpha receptor show impaired transcription of the TGF-beta1 gene and did not display fibrosis. Likewise, it appears that STAT6 plays a role in fibroblast TGF-beta1 transcription after IL-4 or IL-13 stimulation. These findings suggest that an epistatic interaction between IL-4 and TGF-beta may exist which is crucial for pathologic sclerotic activity. PMID- 16380823 TI - Influence of cardiopulmonary bypass on the state of cognitive functions in patients with ischemic heart disease. AB - Forty patients with ischemic heart disease and undergoing aortocoronary shunting surgery with cardiopulmonary bypass were studied. All patients were subjected to neuropsychological assessment and immunochemical analysis of the production of chemokines (IL-8, IP-10, MCP-1, MCP-3, MIP-1 beta, SDF-1 alpha) and cytokines (TNF-alpha and IL-10). The aims of the study were to assess the presence and severity of cognitive deficit developing after surgery with cardiopulmonary bypass and to assess the effects of intraoperative Trasylol on its severity. Cognitive deficit on day 9 after coronary shunting with cardiopulmonary bypass was seen as impairments of attention, hearing-speech memory, visual memory, and dynamic praxis. Trasylol had a marked neuroprotective effect and suppressed the systemic inflammatory response. Patients given intraoperative Trasylol had no clinically significant cognitive deficit in the early post-operative period. PMID- 16380819 TI - Human dendritic cell subsets for vaccination. AB - Protective immunity results from the interplay of antigen (Ag)-nonspecific innate immunity and Ag-specific adaptive immunity. The cells and molecules of the innate system employ non-clonal recognition pathways such as lectins and TLRs. B and T lymphocytes of the adaptive immune system employ clonal receptors recognizing Ag or peptides in a highly specific manner. An essential link between innate and adaptive immunity is provided by dendritic cells (DCs). As a component of the innate immune system, DC organize and transfer information from the outside world to the cells of the adaptive immune system. DC can induce such contrasting states as active immune responsiveness or immunological tolerance. Recent years have brought a wealth of information regarding DC biology and pathophysiology that shows the complexity of this cell system. Thus, presentation of antigen by immature (non-activated) DCs leads to tolerance, whereas mature, antigen-loaded DCs are geared towards the launching of antigen-specific immunity. Furthermore, DCs are composed of multiple subsets with distinct functions at the interface of the innate and adaptive immunity. Our increased understanding of DC pathophysiology will permit their rational manipulation for therapy such as vaccination to improve immunity. PMID- 16380824 TI - Use of p300 cognitive evoked potentials in the diagnosis of impairments of higher mental functions after cardiac surgery in conditions of cardiopulmonary bypass. AB - A total of 40 patients with ischemic heart disease undergoing aortocoronary shunting surgery in conditions of cardiopulmonary bypass were studied. Parameters of the P300 cognitive evoked potentials were studied before surgery and 7-9 days after surgery. Neurological and neuropsychological assessments were also performed. The most significant parameters of the P300 potential were found to be the latencies of the P3 and N2 components, increases in which showed positive correlations with the extent of the developing cognitive deficit. Evidence supporting the neuroprotective effects of Trasylol given during surgery was obtained. Patients given Trasylol showed less marked cognitive deficit and smaller changes in P300 parameters. Analysis of the P300 cognitive evoked potential can be recommended for detecting early cognitive dysfunction and assessing the efficacy of neuroprotective therapy in patients undergoing surgery with cardiopulmonary bypass. PMID- 16380825 TI - Neurophysiological correlates of induced discrete emotions in humans: an individually oriented analysis. AB - Studies on 30 right-handed subjects addressed EEG characteristics (62 channels) in conditions of laboratory simulation of induced emotions of happiness, joy, anger, disgust, fear/anxiety, and sadness. Induced emotions were found to produce, along with common features, individual patterns in the distribution of amplitude-frequency EEG characteristics. Induced positive and negative discrete emotions were characterized by interhemisphere activatory asymmetry in the theta 2 (4-6 Hz), alpha-2 (10-12 Hz), and beta-1 (12-18 Hz) ranges. Experience of the emotions of joy, anger, and disgust occurred on the background of asymmetrical increases in activity in the anterior cortex of the left hemisphere in the theta 2 range, suggesting a leading role for the activity of these areas in realizing the cognitive components of emotional reacting. In addition, some high-ergicity negative emotions evoked combined alpha-2 and beta-1 desynchronization (disgust) or beta-1 desynchronization (fear/anxiety) in the right parietal-temporal cortex, suggesting its involvement in the mechanisms of non-specific emotional activation. These data provide evidence that each of these emotions is characterized by its own individual pattern in the distribution of the amplitude frequency characteristics of the EEG and, on the other hand, that series of ranges and cortical areas show similar but different (in terms of intensity) effects in response to emotional activation for emotions of different flavor. PMID- 16380826 TI - Evoked activity in the hypothalamus and amygdala of the cat in conditions of food related motivation and emotional tension. AB - The amplitude-time characteristics of potentials evoked by clicks were analyzed in bilateral leads from the lateral hypothalamus and amygdala in cats in conditions of food-related motivation, emotional tension (presentation of dogs), and orientational reactions. In conditions of food-related motivation, as compared with the satiated state, there were decreases in the latent periods and changes in the amplitudes of the P1 and N2 components in the hypothalamus and P1, N2, and N3 in the amygdala. The most marked changes occurred on the left side in both structures. Presentation of dogs induced decreases in the latent periods of all components (including N1) of evoked potentials in the hypothalamus and amygdala, the most marked changes in the hypothalamus occurring on the right side and the most marked changes in the amygdala occurring on the left side. Conversely, orientational reactions to emotionally neutral stimuli induced increases in the latent periods of evoked potentials. It is concluded that there is an increase in sensory reactivity in the hypothalamus and amygdala in motivational-emotional states. It is suggested that the side of dominance in these structures may be associated both with the factor of the activity/passivity of the behavior in conditions of fear and the genesis of the emotion (motivational or informational). PMID- 16380827 TI - Involvement of the transmitter systems of the neostriatum in automation of motor skills in dogs. AB - Chronic experiments were performed on six dogs to address the influence of automation of a motor skill on the components of an operant defensive reflex, rearrangement of posture, and the characteristics of the projection of the center of mass on a tensometric platform by the forepaws; the effects of activation and blockade of muscarinic and dopamine receptors on the process of automation of the motor skill were also studied. Despite high criteria for execution of the operant reflex, the reproducibility of reflex performance, increases in the tonic component of the response, and the clear diagonal pattern of the postural rearrangement, a local (as opposed to diffuse) projection of the position of the center of mass on the tensometric platform by the forepaws was seen only after prolonged training leading to automation of the skill. The effect of automation of the operant response could be obtained immediately following bilateral microinjection of the muscarinic receptor agonist carbachol into the neostriatum. The same effect, but weaker, was obtained using bilateral microinjections of the dopamine D2 receptor blocker raclopride into the neostriatum. Conversely, bilateral microinjections of the selective muscarinic M1 receptor blocker pirenzepine into the neostriatum gave the opposite result, with increases in the phasic component of the response, impairment of the diagonal pattern of the postural rearrangement, and diffuseness of the projection of the center of mass on the tensometric platform by the forelimbs. It is concluded that the indirect efferent output of the neostriatum has an important role in the process of learning the motor operant reflex involving maintenance of a defined degree of flexion (where the main role is played by the tonic component of the movement) and during the process of automation of the motor skill. PMID- 16380828 TI - Changes in anxiety levels are followed by changes in behavioral strategy in mice subjected to stress and in the extent of stress-induced analgesia. AB - The experiments reported here demonstrated that corasol increased the extent of analgesia induced by stress and decreased the duration of immobility in mice in a forced swimming test in cold water. Administration of diazepam led to the opposite changes and counteracted the actions of the anxiogen. The effects of the anxiolytic were more apparent in NMRI than mongrel mice, while in mongrel mice the effects of the anxiogen were more marked. Changes in measures following administration of agents were reciprocal in nature. These results lead to the conclusion that that these changes are determined by the level of anxiety, and that the strain differences between mongrel and NMRI mice are also linked with this factor. PMID- 16380829 TI - A functional continuum of regulatory anxiety-enhancing peptides. The search for complexes providing the optimal basis for developing inhibitory therapeutic agents. AB - Regulatory peptides are actively involved in controlling most physiological processes. One such function is regulation of the level of anxiety and panic states. We report here a meta-analysis of data published from 1960 to 2004 on the effects of anxiety-enhancing regulatory peptides. The resulting database was used to investigate the organization and functioning of the anxiogenic regulatory peptide system. Using vector representation of the effects of these peptides, the spectra of physiological effects which might be provoked by each anxiety- and fear-increasing regulatory peptide alone and in combination were evaluated. Complexes of regulatory peptides with anxiogenic profiles with the greatest potential for the further experimental development of inhibitory pharmacological agents were identified. PMID- 16380830 TI - Possible mechanisms of the involvement of dopaminergic cells and cholinergic interneurons in the striatum in the conditioned-reflex selection of motor activity. AB - A possible mechanism for the involvement of cholinergic interneurons in the striatum and dopaminergic cells in the substantia nigra in the selection from among several types of motor activity during learning is proposed. Selection is triggered by simultaneous increases in the activity of dopaminergic neurons and a pause in the activity of cholinergic interneurons in response to the conditioned signal. The appearance of the pause may facilitate activation of GABAergic interneurons in the striatum and the action of dopamine on D2 receptors on cholinergic interneurons. Differently directed changes in dopamine and acetylcholine levels synergistically modulate the efficiency of corticostriatal inputs, such that the rules for modulation of the "strong" and "weak" inputs are opposite in sign. The subsequent reorganization of neuron activity in the cortex basal ganglia-thalamus-cortex circuit leads to increased activity in those cortical neurons providing "strong" innervation to the striatum with simultaneous decreases in the activity of neurons providing "weak" innervation to the striatum, which may underlie the selection of the movement reaction, in which the neocortex is involved. It follows from this model that if the delay between the conditioned and unconditioned stimuli is not longer than the latent period of the reactions of dopaminergic and cholinergic cells (about 100 msec), selection of movement activity in response to the conditioned signal and learning is hindered. PMID- 16380831 TI - Involvement of the motor cortex in the bimanual unloading reaction: a transcranial magnetic stimulation study. AB - The responses of the biceps brachii muscle of the upper arm to magnetic stimulation of the motor cortex during the postural pretuning and forearm unloading tasks were studied in humans. On active unloading, the amplitude of the evoked response decreased in parallel with a decrease in muscle activity. During stationary holding of the load, the muscle response changed in proportion to the load. When, on the background of stationary holding of the load, the other arm took on the same load, the amplitude of the evoked response in the biceps muscle of the arm holding the load decreased without any change in the muscle activity. Passive unloading was accompanied by similar changes in the response evoked by magnetic stimulation as seen with active unloading. The question of whether the decrease in muscle activity (postural pretuning) in active unloading may be associated with both direct corticospinal influences and influences mediated via subcortical structures is discussed. PMID- 16380832 TI - Simulation studies of the role of intracortical inhibition in the formation of sensitivity to cross-shaped figures. AB - The receptive fields of detector neurons for cross-shaped figures in the visual cortex were modeled in conditions of blockade of intracortical inhibition. The tuning of simulated neurons was compared with and without inhibition in the receptive field. In a simulated detector with convergence from two orientation detectors, acute tuning to the cross widened in the absence of inhibition, becoming invariant to the shape and orientation of the cross. A detector based on the disinhibition mechanism lost cross sensitivity when inhibition was blocked and became a detector for the orientation of a single bar. A model of a receptive field in which the inhibitory zones mask the tuning to a cross-shaped figure and in which blockade of inhibition affects only sensitivity is also proposed. We identified which of the properties of receptive field (configuration, location, zone weightings) allow them to simulate the properties of cat visual cortex field 17 neurons, these being sensitive to the shape and orientation of cross-shaped figures. PMID- 16380833 TI - Spike activity of neurons in the amygdala and hypothalamus in bilateral leads in food motivation. AB - Measurements of mean spike frequencies and plotting of autocorrelation histograms were performed to study the nature of the spike activity of individual neurons in the lateral hypothalamus and central nucleus of the amygdala from bilateral leads in rabbits in calm waking, after 24-h food deprivation, and after satiation. The nature of neuron spike activity changed in different ways in the hypothalamus and amygdala on transfer from hunger to satiation: 1) the mean spike frequency changed in more hypothalamic (85%) than amygdalar (56%) cells; 2) as compared with calm waking and satiation, hunger was associated with a decrease in the number of neurons with periodic discharges in the delta frequency range in the hypothalamus, while the amygdala showed an increase in periodic discharges in the beta-2 frequency range; 3) in hunger, the hypothalamus showed a decrease in the number of neurons with burst and periodic discharges, while the amygdala showed increases in the number with equiprobabilistic discharge activity. Assessed in terms of autocorrelation histogram shape, greater changes in the nature of neuron spike activity associated with changes in state occurred on the left side than on the right in both the hypothalamus and amygdala. The maximal differences in neuron spike activity on the right and left sides in the hypothalamus were seen in hunger; the greatest differences in the amygdala were seen in satiation. PMID- 16380834 TI - Protein synthesis-dependent reactivation of a contextual conditioned reflex in the common snail. AB - We report here a study of the effects of blockade of protein synthesis with anisomycin during reactivation of a contextual conditioned reflex in the common snail. The amplitudes of the defensive reactions of snails to standard tactile stimulation before training were identical in two conditions: 1) testing of responses of snails fixed by the shell to a plastic ball floating in water and 2) on the surface of the terrarium glass. After applying electric shocks to the snails' skin for 5 days, a significant difference in responses reflecting the formation of a contextual conditioned reflex was seen in only one of the contexts. Placing trained snails in the same context (reminding) two days after training with simultaneous injection of anisomycin led to significant weakening of training, while control injections of physiological saline produced no such changes. These data suggest that the mechanisms of memory consolidation after training and reminding are not identical. PMID- 16380835 TI - Phase II trial of imatinib (Gleevec) in patients with metastatic renal cell carcinoma. AB - Fourteen patients with metastatic renal cell carcinoma (RCC) were treated on a Phase II trial with imatinib. Eligible patients had histologically confirmed RCC, metastatic and measurable disease by Response Evaluation Criteria in Solid Tumors (RECIST), Karnofsky performance status (KPS) of at least 70%, life expectancy of more than 3 months, and adequate hematological, renal, and liver function. Imatinib was given orally at a dose of 400 mg bid. The most common toxicities were Grade II/III nausea (28%) and Grade II renal insufficiency (14%). All patients had tumor tested by immunohistochemistry (IHC) for KIT protein (CD117, DAKO). One tumor (7%) demonstrated strong, diffuse expression and the rest were negative. No complete or partial responses were observed in 12 evaluable patients treated with imatinib. PMID- 16380837 TI - Comparative analysis of Trichinella spiralis and Trichinella nativa proteins by two-dimensional gel electrophoresis. AB - Trichinella spiralis and Trichinella nativa are both common wildlife parasites in Finland. However, they differ substantially in their resistance to below 0 degrees C temperatures in their natural hosts. T. nativa can live in frozen fox meat for years, whereas T. spiralis dies when frozen. In mouse muscle, the difference is not as evident; even T. nativa cannot maintain infectivity when kept at -20 degrees C for 1 week. Crude larval protein extracts of these two parasite species were analyzed by two-dimensional gel electrophoresis (2DE). The protein patterns showed clear differences, but matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) peptide mass fingerprint followed by database searches failed to identify these proteins, suggesting that they may still be uncharacterized. The patterns compared after freezing treatment at -20 degrees C revealed changes in the intensity of some protein spots. The antigenic differences of the species were analyzed with two-dimensional Western blots, which showed T. spiralis-specific proteins. PMID- 16380838 TI - Serum cytokine levels in human ascariasis and toxocariasis. AB - Cytokine-mediated regulation of chronic intestinal helminth infections is well documented. The present study reports the serum cytokine responses in 38 ascariasis (stool samples positive for Ascaris lumbricoides ova) and toxocariasis (seropositive) patients, 8 ascariasis-positive and toxocariasis-seronegative patients, 22 endemic, normal, healthy subjects residing in areas hyperendemic for ascariasis and 16 normal healthy subjects residing in a low-endemic area in India. The results indicated T-helper type-2-type cytokine responses in ascariasis and toxocariasis (seropositive) and ascariasis-positive and toxocariasis-seronegative patients. The important observation was that both patients and healthy individuals in ascariasis-hyperendemic areas had significantly higher interleukin-5 (IL-5) responses than non-endemic control subjects. The altered immune responses of patients in areas hyperendemic of ascariasis may have further implications. Earlier reports suggest that the geohelminth parasites in endemic areas may modulate the immune response to oral vaccines. A critical role for IL-5 in the immune response against challenge infection consistent with the association of type-2 cytokines with vaccine mediated protection has been reported. Furthermore, co-infection by pathogens that elicit opposing immune responses, particularly helminths vs HIV and tuberculosis, could influence the infection dynamics, progression and immunoprophylaxis of the diseases they cause. Further studies are warranted to ascertain these findings. PMID- 16380839 TI - Raillietiella morenoi sp. n. (Pentastomida) from Gallotia atlantica (Peters and Doria, 1882) (Lacertidae) in the Canary Islands. AB - Raillietiella morenoi sp. n., a new cephalobaenid pentastomid found in the lungs of Gallotia atlantica collected in the Canary Islands, is described. The new species belongs to the sharp-tipped posterior-hook type. The annulus number, morphology, and dimensions of copulatory spicules and the dimensions of anterior and posterior hooks separate Raillietiella sp. n. from the other raillietiedid sharp-tipped posterior-hook species of small lizard parasites in Africa. The host character of endemic protected species of G. atlantica in Alegranza Island posed great difficulty in obtaining more parasite materials. However, more studies are required to state the variability of this species and its possible distribution in other species in the Canary Islands, as well as in other Gallotia spp. PMID- 16380836 TI - From methotrexate to pemetrexed and beyond. A review of the pharmacodynamic and clinical properties of antifolates. PMID- 16380841 TI - The relative importance of olfaction and vision in a diurnal and a nocturnal hawkmoth. AB - Nectar-feeding animals can use vision and olfaction to find rewarding flowers and different species may give different weight to the two sensory modalities. We have studied how a diurnal or nocturnal lifestyle affects the weight given to vision and olfaction. We tested naive hawkmoths of two species in a wind tunnel, presenting an odour source and a visual stimulus. Although the two species belong to the same subfamily of sphingids, the Macroglossinae, their behaviour was quite different. The nocturnal Deilephila elpenor responded preferably to the odour while the diurnal Macroglossum stellatarum strongly favoured the visual stimulus. Since a nocturnal lifestyle is ancestral for sphingids, the diurnal species, M. stellatarum, has evolved from nocturnal moths that primarily used olfaction. During bright daylight visual cues may have became more important than odour. PMID- 16380840 TI - Detection of putative secreted proteins in the plant-parasitic nematode Heterodera schachtii. AB - The beet cyst nematode Heterodera schachtii is an important pathogen worldwide, but its molecular characterization has been limited to studying individual genes of interest. We undertook a high-throughput genomic approach and drastically increased the number of available sequences for this parasite. A total of 2,662 expressed sequence tags were grouped into 1,212 clusters representing a nonredundant catalog of H. schachtii genes. Implementing a bioinformatic workflow, we identified 50 sequences coding for candidate secreted proteins. All of these contain a putative signal peptide required for entry into the secretory pathway and lack any transmembrane domain. Included are previously postulated cell-wall-degrading enzymes and other parasitism-related genes. Moreover, we provide the first report of an arabinogalactan endo-1,4-beta-galactosidase enzyme (EC 3.2.1.89) in animals. As sequence data increase at a rapid rate, developing high-throughput genomic screening is a necessity. The in silico approach described here is an effective way to identify putative secreted proteins and prioritize candidates for further studies. PMID- 16380842 TI - Directionality of the pressure-difference receiver ears in the northern leopard frog, Rana pipiens pipiens. AB - We studied the directional response of the coupled-eardrum system in the northern leopard frog, Rana pipiens pipiens. Eardrum behavior closely approximates a linear time-invariant system, with a highly correlated input-output relationship between the eardrum pressure difference and the eardrum velocity. Variations in the eardrum transfer function at frequencies below 800 Hz indicate the existence of an extratympanic sound transmission pathway which can interfere with eardrum motions. The eardrum velocity was shown to shift in phase as a function of sound incident angle, which was a direct result of the phase-shift of the eardrum pressure difference. We used two laser-Doppler vibrometers to measure the interaural vibration time difference (IVTD) and the interaural vibration amplitude difference (IVAD) between the motions of the two eardrums. The coupled eardrum system enhanced the IVTD and IVAD by a factor of 3 and 3 dB, respectively, when compared to an isolated-eardrum system of the same size. Our findings are consistent with the time-delay sensitivity of other coupled-eardrum systems such as those found in crickets and flies. PMID- 16380843 TI - The phase relationships between the diurnal rhythms in axial length and choroidal thickness and the association with ocular growth rate in chicks. AB - Eyes of young chickens show diurnal oscillations in axial length and choroidal thickness that are out of phase. In eyes responding to myopic defocus induced by prior form deprivation, the two rhythms shift into phase. In order to elucidate the possible role for these rhythms in ocular growth regulation, they were measured under visual conditions that altered ocular growth rate. (1) Form deprivation to myopic defocus. Eyes of chicks were monocularly deprived for 5 days. Diffusers were removed. (2) Myopic defocus to hyperopic defocus. Eyes wore positive lenses for 6 days; lenses were removed. (3) Hyperopic to myopic defocus. Eyes wore negative lenses for 5 days; lenses were removed. Eyes were measured using A-scan ultrasonography at 6-h intervals for 24 h over various cycles. The rhythms shift into phase in eyes slowing their growth in response to myopic defocus in all three conditions. This shift precedes by 1 day the decrease in growth in both lens conditions, and is concomitant with it in recovering eyes. There is a positive correlation between the phase difference and growth rate. In conclusion, there is a consistent association between growth rate and phase relationships of the rhythms in axial elongation and choroidal thickness. PMID- 16380844 TI - Molecular control of serial module formation along the apical-basal axis in the sponge Lubomirskia baicalensis: silicateins, mannose-binding lectin and mago nashi. AB - The freshwater sponge Lubomirskia baicalensis (from Lake Baikal) is characterized by a body plan composed of serial modules which are arranged along an apical basal axis. In shallow water, the sponge occurs only encrusting, while in deeper environment (>3 m), this species forms branches and grows in an arborescent manner. Each module is stabilized by bundles of spined oxeas (amphioxeae spicules). The spicules are surrounded by an organic matrix. cDNAs for structural proteins (silicatein and mannose-binding lectin (MBL)) as well as for one regulatory protein (mago nashi) were isolated from L. baicalensis. Surprisingly the silicatein alpha molecule exists in several, at least four, isoforms (a1 to a4). Expression studies revealed that the steady-state levels of transcripts for the silicateins, the mannose-binding lectin, and mago nashi are highest at the top of the branches, while only very low levels are found in cells at the base. Based on in situ hybridization studies, evidence is presented that the spicule formation (1) starts and is completed inside of the bundles, and (2) occurs together with the mannose-binding lectin from the surfaces of the bundles. The data suggest that the modules are sequentially formed. It is speculated that the expression of the silicateins and the mannose-binding lectin might be (partially) controlled by mago nashi. PMID- 16380845 TI - Deterministic and stochastic features of rhythmic human movement. AB - The dynamics of rhythmic movement has both deterministic and stochastic features. We advocate a recently established analysis method that allows for an unbiased identification of both types of system components. The deterministic components are revealed in terms of drift coefficients and vector fields, while the stochastic components are assessed in terms of diffusion coefficients and ellipse fields. The general principles of the procedure and its application are explained and illustrated using simulated data from known dynamical systems. Subsequently, we exemplify the method's merits in extracting deterministic and stochastic aspects of various instances of rhythmic movement, including tapping, wrist cycling and forearm oscillations. In particular, it is shown how the extracted numerical forms can be analysed to gain insight into the dependence of dynamical properties on experimental conditions. PMID- 16380847 TI - Viper bites: treat or ignore? Review of a series of 99 patients bitten by Vipera aspis in an alpine Swiss area. AB - In a well defined alpine area of Switzerland (Valais, about 300000 inhabitants, tourists included) we studied retrospectively over 32 years, 99 patients bitten by vipers (Vipera aspis, the likely unique species in this area). The annual incidence was estimated at 3/100,000, as in other European countries. The mortality was 0% for the adults. The patients, 72 adults and 17 children (13 years and less), were classified in four groups: grade 0 no envenomation (8%), grade 1 minimal (42%), grade 2 moderate (40%), and grade 3 severe envenomation (10%). The 10 patients of grade 3 showed impressive clinical signs and blood abnormalities, as exemplified by our three most severe cases. Only patients of grade 3 must be treated with antivenom and other intensive treatments, but all patients, even grade 1, especially small children, must be observed for several hours. PMID- 16380848 TI - Management of patients with amyotrophic lateral sclerosis. AB - Standard recommendations for the clinical management of patient with ALS have been edited in recent years. These documents emphasise the importance of patient's autonomy. AIM OF STUDY: To measure how these different recommendations can be applied in the context of a general hospital without a specific ALS clinic. METHODS: Review of medical records of 21 patients with an ALS diagnosis treated by the University Hospitals Geneva who died from 1996-2002. RESULTS: Patients suffered from distressing symptoms during their last hospitalisation. Artificial nutrition was given to 5 patients. Six patients had non invasive ventilation (NIV). Written advance directives were only available in 2 cases. Discussions about theses issues were also conducted late in the evolution of the disease. CONCLUSION: Some discrepancies between our daily practice and the existing recommendations exist, particularly regarding the key issues of artificial nutrition and ventilatory support. PMID- 16380849 TI - Differing definitions of contrast-induced nephropathy. PMID- 16380850 TI - Ultrasonographic assessment of the morphological characteristics of the carotid plaque. AB - Apart from the degree of stenosis, plaque morphology has emerged in recent years as an important contributory factor in stroke risk. Ultrasound studies have shown that hypo- or anechogenic plaques carry a higher risk of cerebrovascular events than echogenic ones. Similarly, heterogeneous plaques presenting a complex pattern of echogenicity in ultrasound have also been more frequently associated with the occurrence of neurological symptoms than homogeneous lesions. Further, most studies determining the surface characteristics in ultrasound have found that ulceration also predicted increased risk of subsequent stroke. These studies are, however, based on visual evaluation using different classification systems and presenting a high variability of intra- and interobserver agreement. A quantitative method using a computerised image analysis of the plaque based on a grey-scale median (GSM) value has recently been developed which allows a more objective and reproducible evaluation of plaque echogenicity. Several studies have also shown that low GSM values are associated with an increased stroke incidence. A stratified GSM analysis determining the GSM in each one millimeter thick stratum of the plaque may represent an additional method of determining different components of unstable plaques, such as thinning of the fibrous cap and the position of the necrotic core near the surface. This article reviews the ultrasonic morphology of the carotid plaque, its clinical prognostic value and correlations with histopathological studies. Recent ultrasound developments in the assessment of plaque echogenicity are also discussed. PMID- 16380851 TI - Illicit methadone injecting during methadone maintenance treatment in a specialised out-patient clinic. AB - AIM: The injection of non-sterile methadone designed for oral consumption is associated with serious health risks. There is only a small number of studies on this topic, with divergent results. The main aim of the present study was to obtain data on the frequency of methadone injecting in a state out-patient clinic specialised in substance use disorders. METHODS: Eighty patients in methadone maintenance treatment were interviewed with a short questionnaire. Mean age was 32 years, 76% were male, mean methadone dose was 55 mg. RESULTS: Twenty-six patients (32%) indicated having injected methadone at least once in their life. Only four patients (5%) reported having injected methadone within the preceding month (mean dose 56 mg). All four had injected additional substances during this month. CONCLUSIONS: Frequency rates appeared low compared with other studies despite a generous take-away policy. The results suggest an association between methadone injecting and a more general tendency to inject substances. PMID- 16380852 TI - Impact of right ventricular size on ECG after percutaneous closure of atrial septal defect with Amplatzer Septal Occluder. AB - OBJECTIVES: To assess ECG changes after percutaneous atrial septal defect (ASD) closure in children with significant left-to-right shunt. METHODS: Analysis of data of 36 consecutive children with an ASD who had successful percutaneous ASD closure with an Amplatzer Septal Occluder. Assessment comprised echocardiography and ECG the day before and after the procedure and at 1, 6 and 12 months follow up. RESULTS: The median age (interquartile range) of children was 7.3 (5.3) years. On the day after the procedure the end diastolic diameter of the right ventricle showed already a diminution (34 (12) mm/m2 before intervention vs. 32 (12) mm/m2). ECG changes were first observed at 1 month follow-up (PR interval before intervention 139 (20) ms vs. 132 (20) ms; QRS duration 88 (18) ms vs. 82 (19) ms) and at 6 months follow-up (QRS axis 77 degrees (33) before intervention vs. 72 degrees (53)). With the exception of the QRS duration, ECG intervals and axis were in a normal range in all patients before the procedure. Median QRS duration normalised at 1 year follow-up (83 (8) ms). CONCLUSION: After transcatheter ASD closure, decrease in right ventricular size began rapidly and was followed by reduction of the QRS duration and PR interval within weeks. Shifting to the left of the QRS axis was observed within 6 months follow-up. This study showed that ECG changes due to right ventricular volume overload can regress and normalise after percutaneous ASD closure in children. PMID- 16380853 TI - Acute medical problems due to Ecstasy use. Case-series of emergency department visits. AB - STUDY AIM: To describe the clinical characteristics of Ecstasy (3,4 Methylenedioxymethamphetamine, MDMA) toxicity. METHODS: Retrospective case-study of 52 self-reported Ecstasy intoxications presenting to our Emergency Department (ED) between January 2001 and December 2003. RESULTS: Most patients ingested Ecstasy together with other substances, including alcohol (51.9%) or other illicit drugs (71.1%). Medical problems leading to ED presentation were collapse or loss of consciousness (36.5%), palpitations (19.2%), dizziness or weakness (15.4%), and anxiety (13.5%). When other drugs were used in combination with Ecstasy the clinical presentation significantly changed. Panic reactions were observed in 4 of 13 patients with cocaine co-use (30.7%), compared to 3 of 39 patients without cocaine use (7.7%). Deep coma was found in 11 of 16 patients with co-use of gamma-hydroxybutyrate (GHB) or opiates (68.8%) but in none of the 36 patients who took Ecstasy without these drugs. Most patients were monitored in the ED. Six patients (11.5%) were transferred to an intensive care unit. Medical complications were severe in five patients and included cardiac arrest, hyperthermia, rhabdomyolysis, disseminated intravascular coagulation, renal insufficiency and liver failure, seizures, and one fatal outcome. CONCLUSIONS: The clinical picture of Ecstasy related problems is complicated by multiple drug ingestion. Co-use of cocaine induces panic reactions. Co-use of GHB or opiates results in depressed levels of consciousness. PMID- 16380854 TI - Absence of clinically relevant effect of caspofungin on cyclosporin pharmacokinetics. PMID- 16380855 TI - Amiodarone after unsuccessful direct-current cardioversion of persistent atrial fibrillation. AB - AIM: To assess the safety and efficacy of amiodarone used after unsuccessful direct current (DC) cardioversion of persistent atrial fibrillation (AF). METHODS: The study group comprised 67 patients (F/M 26/41; mean age 61.3+/-11.2 years) after unsuccessful DC cardioversion (DCC) of persistent AF (mean arrhythmia duration 212.6+/-135.2 days) in whom another attempt of DCC was intended. Repeat DC cardioversion was performed after loading with oral amiodarone, for a period necessary to achieve a cumulative dose of up to 12.0 16.0 g. Pretreatment was an outpatient procedure. After successful DC cardioversion all study subjects received a maintenance dose of amiodarone, 100 200 mg daily, aimed at preventing AF. The follow-up period was 12 months. RESULTS: Spontaneous conversion to sinus rhythm (SR) during amiodarone pretreatment was observed in 13 pts (19.2%). DCC was performed in 54 pts and SR was restored in 41 of the study pts (76%). Complications occurred in 3 pts, including 1 case of apparent hyperthyroidism and 2 cases of decreased TSH level, and required amiodarone withdrawal. After 12 months, 72.2% of pts maintained SR on low dose (179.2+/-42.1 mg/day) amiodarone. Spontaneous conversion to SR during amiodarone loading was significantly related to long-term SR maintenance after successful DC cardioversion (p<0.013; RR 2.01; 95% CI 1.34-3.03). CONCLUSION: Pretreatment with amiodarone and repeat DC cardioversion results in sinus rhythm restoration in about 80.6% of pts with persistent AF after an initial unsuccessful attempt. Direct-current cardioversion can be performed safely taking standard precautions for patients receiving amiodarone. At 12 months after successful repeated DC cardioversion, more than 72.2% of pts on low-dose amiodarone maintain SR. PMID- 16380857 TI - Closure of perimembranous ventricular septal defect using transcatheter technique versus surgical repair. AB - INTRODUCTION: Since the Membr VSD occluders have been introduced into clinical practice it is now possible to compare two treatment methods -- surgical and percutaneous. AIM: Assessment of the effectiveness and risk of complications in patients with perimembranous ventricular septal defect (VSD) treated with the Membr VSD occluder or surgery as well as comparison of postprocedural left and right ventricular systolic functions. METHODS: The study involved 11 children with perimembranous VSD treated with occluder implantation (Group A) and 12 children with surgical repair (Group C). Groups A and C differed slightly in terms of age of patients (p=0.026), but had similar mean weights (p=0.103), pulmonary to systemic flow ratios (Qp: Qs, p=0.929) and follow-up duration after the procedure. No significant differences were observed between the two groups in terms of left ventricular systolic function [left ventricular ejection fraction (EF, p=0.567)], diastolic function [mitral flow E/A ratio (p=0.975)], E deceleration time (DCT, p=0.346), isovolumetric relaxation time (IVRT, p=0.606), heart rate (HR, p=0.133) or left ventricular diameter (LV) on transthoracic echocardiography (TTE) before VSD closure. TEE was performed in all patients and the parameters listed above were measured within 3 to 16 months after the procedure, and then intra- and inter-group comparisons were carried out. Additionally, mitral (MV), tricuspid (TV) and aortic (Ao) valve regurgitations were evaluated. RESULTS: Tricuspid (TR, p=0.028) and mitral regurgitation (MR, p=0.043) decreased significantly after closure of VSD with the Membr VSDO. MR was significantly smaller in patients treated with the occluder than in those after surgery (p=0.026). Postprocedural TR (p=0.486) and aortic regurgitation (AR, 0.607) did not differ significantly between the two groups. Left ventricular EF was significantly (p=0.004) lower and HR higher (0.043) after surgery than in children treated with the occluder. No significant differences of the diastolic function were found - E/A (p=0.88), DCT (p=0.413), IVRT (p=0.09). CONCLUSIONS: 1. During the mid-term follow-up left ventricular EF was higher in patients after Membr VSDO occluder implantation than after surgical repair of VSD. 2. Higher EF and reduction of the left ventricular diameter after closure of perimembranous VSD with the occluder may result in a lower incidence and degree of MR in comparison to the subjects operated on. PMID- 16380859 TI - Heart rate variability and left ventricular mass in slim children and young adults with hypertension. AB - INTRODUCTION: Uncontrolled arterial hypertension brings direct and long-term sequelae in adult age, such as stroke, ischaemic heart disease with myocardial infarction, left ventricular hypertrophy or cardiac arrhythmia. AIM: To assess heart rate variability (HRV) spectral parameters and left ventricular mass in slim children with arterial hypertension, and to search for correlations between these two parameters. METHODS: 35 children aged 14.4+/-3.1 with idiopathic untreated arterial hypertension were enrolled. The control group included 30 age- and gender-matched healthy children (aged 14.1+/-2.9 years). In all analysed subjects an analysis of HRV parameters (high frequency (HF) and low frequency (LF) components) during 10-minute waking state and sleeping time was performed and left ventricular mass (LVM) as well as the left ventricular mass index (LVMI, g/m) were assessed based on echocardiographic measurements. RESULTS: There was no difference in LF during the waking state and sleep HF between the two groups, whereas HF values during the waking state were significantly lower (p<0.05) in children with hypertension. The LF/HF index from both registration intervals was significantly higher in the group of children with hypertension. In children with hypertension, LVM and LVMI correlated significantly with LF (r=0.32, p<0.05 and r=0.39, p<0.01). LVM and LVMI correlated positively with the LF/HF index during night hours (r=0.45, p<0.004 and r=0.49, p<0.002). No significant correlations were found between the analysed parameters in children from the control group. CONCLUSIONS: The increase of sympathetic activity during sleep correlates significantly with left ventricular mass and corrected left ventricular mass index in children with arterial hypertension. PMID- 16380861 TI - The prognostic significance of early dobutamine echocardiography in patients with acute myocardial infarction treated with primary coronary angioplasty. AB - BACKGROUND: The prognostic significance of early dobutamine echocardiography (DE) after successfully treated acute myocardial infarction (AMI) with primary coronary angioplasty (PTCA) is still unclear. Patients who respond to DE may have better left ventricular function improvement and possibly a better clinical outcome. AIM: To assess whether early DE can predict spontaneous functional recovery in patients treated successfully with primary PTCA and whether responders to DE have a better clinical outcome. METHODS: DE (5 and 10 ug/kg/min) was performed in 110 consecutive patients (61+/-10 years) 4+/-1 days after successful primary PTCA (TIMI 3, stenosis <30%). Left ventricular ejection fraction (LVEF) and wall motion index (WMSI) were measured. Patients underwent clinical assessment and two-dimensional echocardiography at 3 and 6 months. RESULTS: In the DE responders (76 pts), LVEF increased significantly from 41%+/ 9% at baseline to 47%+/-10% at 6 months (p<0.0001), whereas the improvement found in nonresponders (34 pts) was insignificant (from 36.3%+/-9% at baseline to 38.8%+/-10% at 6 months, p=0.4). The nonresponders to DE had a higher incidence of subsequent revascularisation (4/34 (11.8%) vs 3/76 (3.9%) p=0.12), reinfarction (5/34 (14.7%) vs 2/76 (2.6%), p=0.28) and death (3/34 (9%) vs 0/76 (0%), p=0.0086). The incidence of combined end-point (revascularisation, reinfarction and death) was significantly lower in the group of responders to early DE (p=0.03). CONCLUSIONS: Early DE can precisely predict functional recovery and the extent of irreversibly damaged myocardium in patients with AMI in whom anterograde flow is fully restored. A positive response to early DE is associated with a better clinical outcome and prognosis. PMID- 16380863 TI - Ventricular activation patterns during different pacing modes. An insight from electroanatomical mapping. AB - BACKGROUND: Biventricular pacing (BiV) is employed as the current standard for cardiac resynchronisation therapy. Other pacing modalities have been proposed as alternatives; however, data on changes in electrical activation sequence caused by pacing from various sites are limited. AIM: To describe changes in activation patterns during different ventricular pacing modes in patients with chronic heart failure. METHODS: A total number of 20 patients (mean age 59.6+/-8 years) with chronic heart failure, intraventricular conduction abnormality (QRS >130 ms) or complete AV block were studied. Endocardial activation maps of both ventricles (CARTOTM, Biosense-Webster) were obtained during spontaneous rhythm and biventricular (BiV, n=9), right ventricular bifocal (BiF, n=7) and single-site left ventricular (LV, n=4) pacing. The following parameters were assessed: activation pattern, total LV endocardial activation time (LVAT) and electrical interventricular delay (IVD). RESULTS: Right ventricular apical pacing was associated with the longest LVAT (145+/-24 ms). On the contrary, both BiV and BiF pacing shortened LVAT with BiV being superior in the degree of LVAT reduction (89+/-13 vs 103+/-10 ms, p<0.05). BiV pacing also significantly shortened IVD and modified the LV activation sequence in a complex manner. Such changes were not observed during BiF pacing. In the presence of fusion with spontaneous activation, single-site LV pacing was comparable to BiV pacing. CONCLUSIONS: Among the different pacing modes, BiV pacing and single-site LV pacing with fusion resulted in the most pronounced changes in ventricular activation that appear to be a prerequisite for successful resynchronization of both ventricles. PMID- 16380865 TI - Carotid and femoral atherosclerotic lesions in patients with coronary heart disease confirmed by angiography. AB - INTRODUCTION: Clinically evident atherosclerosis is preceded by preclinical changes in the arterial wall. These changes are characterised by increased thickness of the intima-media complex (IMT). AIMS: A complex ultrasound assessment of the peripheral vessels as well as an attempt to find ultrasound parameters correlating with the burden of atherosclerotic lesions of the coronary arteries. METHODS: 231 patients who underwent both coronary angiography and ultrasound examination of the following arteries: common carotid artery (CCA), carotid bulb and common femoral artery (CFA) were enrolled. The IMT value, presence of plaque and Doppler blood flow parameters were evaluated. Selected clinical and biochemical risk factors of atherosclerosis were assessed. Two groups of patients were analysed: 200 patients with coronary artery disease confirmed by angiography (study group), and 31 patients with normal coronary arteries (control group). RESULTS: Significantly higher values of the IMT in the peripheral arteries were observed in patients with coronary artery lesions than in those without (CCA - 0.91 vs 0.61 mm, carotid bulb -- 1.31 vs 0.67 mm, CFA -- 1.38 vs 0.63 respectively, p<0.0001). Atherosclerotic plaques were present only in patients with coronary artery disease. Additionally, IMT values of the CCA, carotid bulb and CFA were significantly higher in patients with severe coronary artery disease (three vessel disease) than in patients with lesions in one or two coronary arteries. CONCLUSIONS: Patients with coronary lesions present with increased IMT values and higher plaque occurrence. Complex ultrasound evaluation of different peripheral arteries (CCA, carotid bulb and CFA) may be used as part of the cardiovascular risk stratification. PMID- 16380867 TI - [Right atrial mobile thrombus in a patient with acute, massive pulmonary embolism effectively treated with thrombolysis -- a case report]. AB - A 74-year old woman was admitted to the orthopaedic word due to femoral bone fracture. Six days later she was admitted to the intensive care unit because of the clinical signs of circulatory and respiratory failure. Because acute pulmonary embolism was suspected, transthoracic echocardiography was performed and showed typical signs of acute massive pulmonary embolism as well as a highly mobile, longitudinal, additional echo in the right atrium attached to the interatrial septum in the place of foramen ovale, prolapsing in the diastole into the right ventricle. The patient was immediately and successfully treated with thrombolysis. Treatment options in patients with acute pulmonary embolism and right atrial mobile thrombus are discussed. PMID- 16380869 TI - [Ultrasonographic contrast injection enhances safety of thrombin - induced obliteration of femoral artery pseudoaneurysm]. AB - Thrombin injection is known as an efficient and safe method of femoral artery pseudoaneurysms treatment, however, it can be complicated by peripheral thrombosis to accidental casual thrombin injection into femoral artery. We present our own procedure modification. Before thrombin injection we inject into pseudoaneurysm a small volume of ultrasonographic contrast. Doppler signal enhancement confirms proper niddle position. PMID- 16380871 TI - [Bronchial adenocarcinoma mimicking acute pulmonary embolism -- a case report]. AB - A case of a 31-year-old male with medical history, clinical symptoms and additional diagnostic tests strongly suggesting diagnosis of acute pulmonary embolism is presented. However, pulmonary emboli were not seen during pulmonary angiography. At autopsy bronchial carcinoma was found and this definitive diagnosis was confirmed by the results of histopathological study. PMID- 16380872 TI - [Wellens's syndrome -- the ominous T-waves]. AB - We describe three patients with Wellens's syndrome: biphasic T-waves or deeply inverted T-waves in the precordial leads plus a history of acute coronary syndrome without serum marker abnormalities. Wellens's syndrome is due to critical stenosis of the proximal left anterior descending artery and there is a high risk for anterior wall myocardial infarction and/or sudden cardiac death within a few weeks. Early coronary angiography with subsequent PCI or CABG should be performed in these patients. The T-wave changes usually occur during a pain free interval but resolve within 12 months after revascularisation. PMID- 16380874 TI - [Double-orifice mitral valve -- case report]. AB - A case of 16 year old male with an initial diagnosis of the mitral stenosis is presented. The transthoracic echocardiography showed the complete bridge type double-orifice mitral valve with a larger medial orifice. PMID- 16380875 TI - [Recurrent myocardial infarction leading to heart failure in a patient with a history of previous infarction successfully treated by primary angioplasty]. PMID- 16380878 TI - [Do we know the mechanisms of favourable effects of n-3 unsaturated fatty acids?]. PMID- 16380877 TI - [Hypotensive treatment after stroke -- the MOSES study documented superiority of eprosartan over nitrendipine]. PMID- 16380879 TI - [Is the use of vasodilator therapy justified in patients with asymptomatic, significant aortic regurgitation?]. PMID- 16380880 TI - [RF ablation of four arrhythmias associated with the presence of pseudo-Mahaim fibres -- a case report]. PMID- 16380881 TI - [Assessment of endothelial function and carotid intima-media thickness: clinical applications]. PMID- 16380882 TI - [Genetic diseases of the eye]. AB - The eye has always provided a diagnostic window for hereditary and acquired systemic diseases. Of the more than 6,000 known hereditary diseases, many are associated with changes of the visual system. Some are isolated genetic diseases of the eye, others are associated with additional ocular or systemic disorders (syndromes). In recent years, the recognition of genetic diseases as a leading cause of severe visual impairment in adults and in children has led to efforts to determine the underlying defects as well as to develop diagnostic and therapeutic molecular genetic tools. Also, education of ophthalmologists about these diseases will foster the prevention of and therapeutic approaches for genetic blindness. In this article, current knowledge on the clinical manifestations, aetiology and management of genetic diseases of the eye has been summarised. PMID- 16380883 TI - [Does the orientation of the IOL in the eye play a role? Imaging quality of foldable intraocular lenses measured utilizing point spread function (PSF measurement) and scanning electron microscopy cross-sectional images]. AB - BACKGROUND: Foldable intraocular lenses (IOLs) are established in ophthalmic surgery and increasingly more complex geometries are available. This study investigates different designs of the optics of the IOL as well as the imaging quality of these optics utilizing electron microscopic photographs and mathematical approximations. METHOD: Included in this study were two different lens powers of common IOLs (+ 13 D and + 26 D). All studied IOLs were processed depending upon their water content: Aqueous IOLs with up to 25 % water content were fixed with 2.5 % glutaraldehyde in 0.1 M phosphate buffer (mPP), and then contrasted with 2 % OsO (4) and processed in an alcohol series of 30 - 100 % propanol. Subsequently, the IOLs were transferred into EPON (epoxy resin) and polymerization took place at 60 degrees C. After hardening, the samples were cut out with a leaves saw and divided into halves. Afterwards these were polished with glass and diamond knifes at the ultramicrotom (Ultracut, Ruckert-Jung). After coal vaporization measurement took place in the scanning electron microscope (DSM 962, Zeiss). In the case of dry stored IOLs, the samples were sputtered with Au-Pd 20 nm and afterwards measurement was performed as described above. The measurement of the optical imaging quality was performed with correctly and inversely oriented IOLs according to the standard EN ISO 11979 -- 2 by means of a measuring system which works on the basis of the point spread function (PSF) and/or the modulation transfer function (MTF). From this the Strehl ratio was computed in each case. RESULTS: Utilizing a correlation of the electron microscopic investigation of the optics of different IOLs with their imaging quality (Strehl ratio), we were able to show that the optical design of the IOL has substantial influence on the imaging quality of an intraocular lens. In particular, with non-symmetrical designs of the optics (e. g., AR40 e; AMO) the orientation of an IOL in the eye is crucial; i. e., the Strehl ratio increased thereby from 0.68 to 0.87. CONCLUSIONS: The design of asymmetrical optics is crucial and much attention should be paid to the fact that the imaging quality can substantially be affected. A correct orientation of the lens within the eye is very important with these asymmetrical optics. PMID- 16380884 TI - [Asphericity of the cornea and astigmatism]. AB - BACKGROUND: Regarding astigmatism of the cornea, curvature as well as asphericity depend on the meridional axis. Their functional dependence and relation to the quality of the retinal image are still unclear. METHODS: The astigmatic eye was modelled using a biconoid anterior corneal surface by means of a commercially available optical designer programme (Zemax EE, Zemax). The influence of asphericity and astigmatism on the quality of the retinal image was determined by means of ray tracing. Thirty eyes with astigmatism of up to 5.3 D underwent corneal topography (Keratograph C, Oculus) which allowed a numerical evaluation of the cylindric power as well as the asphericity in the main meridians. RESULTS: The quality of the retinal image of an eye with corneal astigmatism can be improved by a factor of 2.28 if the asphericity is optimised. Correction of the central astigmatism only (without considering asphericity) yields only a rather marginal improvement. The average difference of the asphericity in the main meridians is close to zero, however, the individual difference ranges from - 0.372 to + 0.444 which is definitely clinically relevant. CONCLUSIONS: Anisotropic asphericity of the cornea may significantly enhance or compensate central corneal astigmatism. Clinically manifest astigmatism is an individually variable combination of asphericity and curvature difference in the two main meridians and is dependent on the pupil size. Laser correction of corneal astigmatism must take meridional asphericity into account. PMID- 16380885 TI - [Accuracy of the autorefractor power refractor in clinical work--a comparative study]. AB - BACKGROUND: The aim of this prospective study was to investigate accuracy and efficiency of the autorefractometer PowerRefractor compared to established autorefractometers and retinoscopy as standard method. PATIENTS AND METHODS: 150 patients (300 eyes) were examined. Patients were initially (pupil not influenced) analyzed with the Power Refractor (PR (neutral)). After that a examination followed under standardized Cyclopentolat cycloplegia with the Power Refractor (PR (cyclo)), the hand-held Retinomax K-plus (Nikon), the table-top mounted RK-5 (Canon) and retinoscopy. RESULTS: Deviation from retinoscopy (percentage of values with more than 1 dpt difference/maximal deviation): Sphere: PR (neutral): 66.3 %/8.5 dpt; PR (cyclo): 48 %/5 dpt; RK-5: 26.7 %/4.75 dpt; Retinomax K-plus: 6.7 %/1.25 dpt; Cylinder: PR (neutral): 12 %/6.25 dpt; PR (cyclo): 21 %/4.75 dpt; RK-5: 1.3 %/2.75 dpt; Retinomax K-plus: 2.3 %/2.5 dpt; spherical equivalent: PR (neutral): 44.7 %/7.1 dpt; PR (cyclo): 35.3 %/5.4 dpt; RK-5: 9 %/4.5 dpt; Retinomax K-plus: 1.3 %/1.,4 dpt; error of entire refraction: PR (neutral): 62.7 %/7.8 dpt; PR (cyclo): 61.3 %/7.3 dpt; RK-5: 24 %/4,5 dpt; Retinomax K-plus: 14.7 %/3.6 dpt CONCLUSIONS: The best accuracy compared to retinoscopy had Retinomax K plus followed by RK-5. PowerRefractor had the biggest deviations of the tested devices. PowerRefractor may be a usefull device for screening small children and handicapped people because of the one meter observing distance. For a precise refraction especially in children a retinoscopy under cycloplegic conditions is still necessary. PMID- 16380886 TI - [Driving habits of patients with glaucoma]. AB - BACKGROUND: Central visual field defects due to glaucoma are common, increasing with old age. Impaired visual processing, for instance caused by glaucoma, may play a role in the aetiology of car accidents involving older drivers which can result in personal injury. Mandatory eye exams with assessment of the visual field in elderly people holding a driving licence will become more and more important, especially in a continuously ageing and increasingly mobile population. MATERIALS AND METHODS: In this prospective study, 80 patients with overt glaucoma and 52 patients without glaucoma, all holders of a valid driving licence, were enrolled. For each patient, the best corrected visual acuity was recorded and an examination of the central visual field was performed with automatic perimetry. In addition, a detailed questionnaire about the current driving habits of the patient was requested. RESULTS: In summary, 29 patients (36 %; 95 % CI: 26 - 48 %) of 80 glaucoma patients were driving a motor vehicle with binocular congruent scotomata within the central 30 degrees visual field, which is not sufficient to meet current legal requirements in Austria. In addition, 3 out of 29 impaired patients had a visual acuity that was below the mandatory legal requirements. A total of 39 patients (49 %; 95 % CI: 37 - 60 %) of the glaucoma patients fulfilled legal requirements. Examination of these patients showed only monocular or binocular central visual field defects that were not congruent. However, 12 (15 %; 95 % CI: 8 - 25 %) patients were holders of a valid driving licence, but had stopped driving some time ago. Based on the prevalence of glaucoma and the number of driving licence holders, the projected number of actively driving glaucoma patients who do not meet the legal requirements regarding the visual field is probably around 15,400 (7,400 - 29,500) in Austria and around 163,500 (79,000 - 313,500) in Germany. CONCLUSIONS: Time limits for the validity of the driving licence within the European Community have been set. In addition, the legal requirements for driving a motor vehicle should also be clearly defined, especially the requirements regarding the visual field and the acceptable dimensions of central scotomata. In addition, a mandatory eye exam for older drivers to be performed by ophthalmologists should be considered in order to detect persons posing a safety risk in traffic. PMID- 16380887 TI - [Changes in the G-DRG System from 2003 to 2005 in Ophthalmology--Ophthalmologic G DRG 2003 to 2005]. AB - BACKGROUND: The objective of the study was to illustrate the effect of the extensive changes of the German DRG System on reimbursement of clinical ophthalmology during the years 2003, 2004 and 2005. METHODS: All ophthalmologic patients treated as in-patients at the Department of Ophthalmology at the University Munich during the year 2003 served as a reference data basis. By means of appropriate software those cases were then re-grouped according to the G-DRG classification of the years 2003, 2004 and 2005. This resulted in different reimbursements caused only by system changes for an exemplary hospital of maximum medical care. In addition, the same calculations were performed for four virtual, typical clinics based on the calculation data of the "Institut fur Entgeltsysteme (InEK)". For those four clinics it was assumed that 80 % of the cases came from one of the subspecialties retina, glaucoma, cataract or strabismus surgery. RESULTS: Changes in the G-DRG system caused the sample hospital of maximum care to loose 8.5 % case mix index (CMI) during the period of 2003 to 2005. For three of the four virtual, typical ophthalmological clinics the theoretic reimbursement conditions also deteriorated: retina surgery -- 10.6 %, glaucoma surgery - 15.8 % and cataract surgery -- 17.9 % CMI. Only strabismus surgery showed an increase of + 5.6 % in CMI during the period examined. CONCLUSION: Over the years 2003 to 2005 the CMI clearly deteriorated for many ophthalmological subspecialties given otherwise identical conditions. To calculate the changes specifically for an individual hospital, the individual base rates have to be considered. PMID- 16380888 TI - [Bilateral lid swelling during infectious mononucleosis (Hoagland-sign)]. AB - BACKGROUND: Infectious mononucleosis ("kissing disease") is an infection caused by Epstein-Barr-Virus (EBV, human herpes virus 4). METHODS: We present the case of an 18-year old female patient with dacryoadenitis and sub-febrile temperatures followed by a distinct feeling of sickness. RESULTS: Laboratory results revealed a Leukocytosis of 16.3 G/I with explicit elevation of hepatic parameters. The ultrasound scan of the orbital region showed a bilateral, echo-reduced, compressible lesion in the area of the lachrymal gland. The abdomen-ultrasound scan revealed a splenomegaly and the size of the liver was within normal limits. Serology was positive for EBV-VCA-IgG, EBV-EA-IgG and EBV-VCA-IgM. CONCLUSIONS: A dacryoadenitis can be the presenting symptom of an infectious mononucleosis. PMID- 16380889 TI - [Molecular genetic analysis of the BIGH3 gene in lattice type I (Biber-Haab Dimmer) and granular type II (Avellino) corneal dystrophy: is indirect mutation analysis for hot spots recommended?]. AB - BACKGROUND: Mutations of the BIGH3 gene were delineated as the underlying gene defect for corneal dystrophy Lattice Type I (CDL1) and corneal dystrophy Avellino type (CDA) in families with different regional provenance. Missense mutations in exon 4 with single base pair substitution which result in amino acid alterations Arg124Cys (CDL1) and ARG124His are described as hot spots. We report on histopathological and molecular genetic investigations in 2 German families and a single patient with CDL1 and CDA. METHOD: In 3 affected family members and 1 unaffected family member and in one single patient with CDL1 and in 3 affected family members and 1 unaffected family member of a family with CDA mutation analysis in exon 4 of BIGH3 gene by direct sequencing of genomic DNA from peripheral blood was performed. Histopathological examination of corneal tissue of both index patients was performed after penetrating keratoplasty. RESULTS: We revealed a heterozygous single base pair substitution 417C-->T in family A and patient B (CDL1) and a heterozygous single base pair substitution 418G-->A in family C (CDA). In all index patient's diagnosis was confirmed by histopathological examination of corneal tissue. The sequencing results were confirmed by restriction digestion with HpyCH4V (NEB; CDL1) restriction endonuclease site and AvaII (NEB; CDA) restriction endonuclease site. The heterozygous 417C-->T transition in family A and patient B alters the amino acid sequence from Arg124Cys while the heterozygous 418G-->A transition in family C alters the amino acid sequence from Arg124His in the keratoepithelin. COMMENT: Codon 124 of the BIGH3 gene appears as a mutation hot spot also in German families with CDL1 and CDA. Indirect mutation analysis with restriction digestion is suggested as first step investigation in families with relevant corneal dystrophies. Direct sequencing of all exons is recommended as a second step if there are no results in restriction digestion. PMID- 16380890 TI - [Spondylosis, spondylolisthesis, spondyloptosis]. PMID- 16380891 TI - [Optimization of cup positioning in THA--comparison between conventional mechanical instrumentation and computer-assisted implanted cups by using the orthopilot navigation system]. AB - AIM: The goal of the current study was to determine the accuracy of free-hand and computer-assisted cup replacement. MATERIAL AND METHODS: We analysed the cup positions after 50 conventional instrumented THAs and compared them with the positions of 50 navigated cups. The operations were done between October 2002 and November 2004. To determine the accuracy of the cup position, the author developed a special measurement tool. With the help of the method it was ensured that the radiograph central beam reached the pelvis in a horizontal position. The central beam was focused directly on the head. We calculated the anteversion and the inclination. RESULTS: The variability of the cup position was significantly higher in the free-hand implanted group. The standard deviation for antetorsion in the navigation group was 5.0 and for inclination angles 2.8. In the conventionally implanted group the standard deviation was 7.4 for inclination and 6.9 for antetorsion. In particular, the anteversion cannot be exactly calculated without a navigation tool since the individual position of the patient's pelvis cannot be assessed by the surgeon without aids. Clinical long-term studies for the clarification of a higher stand-time of the computer-navigated cups are still necessary. PMID- 16380892 TI - [Bilateral hip joint replacement as a one-stage or two-stage procedure for dysplastic coxarthritis: a comparative analysis of 30 patients]. AB - AIM: Dysplastic coxarthritis usually occurs earlier than primary coxarthritis. As a rule dysplastic coxarthritis mostly affects young and active individuals. Bilateral coxarthritis may lead to severe impairment requiring surgical replacement of both hip joints, as the patients are frequently dissatisfied with the substitution of one joint alone. METHOD: In a retrospective study, 15 patients with bilateral hip dysplasia who had received total endoprostheses in both hip joints in a one-stage procedure (group A) were compared to 15 patients with bilateral dysplasia who had been operated on bilaterally in two stages (group B). Group B was subdivided into B1 (first side) and B2 (second side). All dysplasias were classified according to Hartofilakidis and Crowe. RESULTS: On the first postoperative day, group A experienced a mean drop of 33.9% in haemoglobin values, group B1 a mean drop of 25.1% and group B2 a mean drop of 26.3%. Patients of groups A/B1/B2 received on average 1.7/1.1/1.3 units of autologous blood and 1.2/0.1/0.3 units of foreign blood, respectively. Both groups experienced an improvement in their quality of life due to marked improvements in their Harris Hip Score and UCLA Score. Group A had no higher rate of complications. CONCLUSION: Although the single-stage procedure involved greater effort in terms of physiotherapy, patients preferred bilateral implantation of total hip endoprosthesis in a single surgical session because they needed to undergo the process of operation, mobilisation and rehabilitation only once. PMID- 16380893 TI - [Clinical and radiological results of the thrust plate prosthesis in patients with aseptic necrosis of the femoral head]. AB - AIM: In this study, we investigated the results after implantation of the thrust plate prosthesis (TPP) in patients with femoral head necrosis. We intended to answer the question if the femoral neck prosthesis, inaugurated by Huggler and Jacob in Switzerland, which needs a good bone stock for a successful implantation, is a recommendable alternative to other cementless intramedullary fixed prostheses. METHOD: In a prospective study, 62 patients who had received 70 TPP because of femoral head necrosis as a result of various aetiologies between 1993 and 2004 were examined clinically and radiologically. The follow-up examinations were carried out 3 and 6 months postoperatively and subsequently once a year. The mean follow-up interval was 6.0 +/- 1.9 years (1.0-10.2 years). Clinical examination was carried out using the Harris hip score; the radiological examination was performed according to predefined criteria in an exact a.-p. view. Four sectors can be distinguished: A corresponds to the femoral neck stump, B to the bony stock cranial (= 1) and caudal (= 2) to the mandrel of the prosthesis, C is the region above and under the bolt and D is the cortical area around the lateral plate. Furthermore, we performed a Kaplan-Meier survival rate analysis. RESULTS: We found excellent clinical results. The preoperative Harris score increased from 48.3 to 91.6 +/- 6.6 at 24 months after the operation. Radiolucencies of various relevancies were detected depending on the localisation. We often saw bony atrophy under the thrust plate (sector A: A 1 21.4%, A 2 9.9%) and small radiolucencies along the bolt (sector C 30 % in general). As a pathological finding we interpreted progressive radiolucencies of sector B, which was considered to be a sign of loosening, when they showed a thickness of > or = 2 mm. Therefore, we had to change one TPP. In two additional cases we saw an extended atrophy under the lateral plate, which was also interpreted to be a sign of loosening. The Kaplan-Meier survivorship analysis for 5 and 10 years was 95.1% (95% - 0.95 +/- 0.05). Our study suggests that, in spite of a slightly higher aseptic loosening rate in comparison with cementless stem prosthesis, the thrust plate prosthesis proved worthwhile. On account of our previous experience we consider the TPP to be a good alternative implant, especially for young patients. CONCLUSION: Due to excellent clinical results and nearly identical findings in the survivorship analysis in comparison to cementless stem prostheses, the TPP is also a good alternative implant for total hip arthroplasty in patients with femoral head necrosis. For a successful implantation of the TPP a good bone quality of the proximal femur is necessary. Therefore we cannot recommend the use of a thrust plate prosthesis in patients with femoral head necrosis and simultaneous osteopeny. PMID- 16380894 TI - [Prevention of heterotopic ossification after THA with indomethacin: analysis of risk factors]. AB - AIM: Several risk factors are associated with heterotopic bone formation following total hip replacement. All these risk factors were defined in cohorts without any treatment against postoperative ectopic bone. The aim of this prospective study was to reveal risk factors for the development of postoperative ossifications in patients who underwent a 7-day course of indomethacin therapy. METHODS: 211 consecutive patients with 217 cementless total hip arthroplasties were included. Patients were given 100 mg indomethacin daily in a 7-day course. RESULTS: Grade 2 or 3 ossifications were observed in 13 (5%) of the male and 3 (2%) of the female Individuals postoperatively (p = 0.0043; odds ratio = 0.45). Older individuals (p = 0.0021; odds ratio = 1.03) as well as patients with primary osteoarthritis (p = 0.0307; odds ratio = 0.28) also showed a higher risk for developing ectopic bone formations. CONCLUSION: With a 7-day course of indomethacin after total hip arthroplasty, male and elderly individuals, as well as patients with primary osteoarthritis, are considered to be at high risk to develop heterotopic ossifications postoperatively. Indomethacin reduces the incidence of postoperative ectopic ossifications, but not the patient's risk. PMID- 16380895 TI - [Macerated human hip bone preparation as a substitute for fresh frozen preparations for testing the primary stability and frictional torques of cementless hip cups]. AB - AIM: The aim of the study was to validate macerated human acetabuli as replacement for fresh frozen preparations for testing primary stability and the screwing in moments of cementless threaded hip cups. METHOD: Three fresh frozen human pelvis were tested. One half of each pelvis was macerated whereas the other half was preserved as fresh frozen preparation. In the side of every pelvis the moments of screwing-in, the micromotions, the maximum expressing force and the maximum pull-out torque were determined. RESULTS: The screwing in moments, the maximum expressing forces and the maximum pull-out torques did not change. The micromotions were reduced to half. CONCLUSION: Considering the reduction of the micromotions, macerated human acetabuli are valid replacements for fresh frozen preparations for testing the primary stability and the screwing-in behaviour of screwed pans. PMID- 16380896 TI - [Improved osseointegration and periprosthetic bone volume around cementless metal implants under bisphosphonate treatment]. AB - AIM: The effects of a systemic treatment with the bisphosphonate ibandronate on osseointegration of uncoated and hydroxyapatite-coated titanium implants and on periprosthetic bone volume have been evaluated and the dosage of medication had to be defined. METHOD: We used an animal model of the rat, the animals were assigned to three treatment groups receiving 1 microg, 5 microg and 25 microg/kg body weight and one control group receiving NaCl 0.9%. An uncoated and a hydroxyapatite-coated titanium rod were inserted into the medullary canal of the femur. After 28 days the specimens were harvested and histomorphometric evaluation revealed extend of osseointegrated implant surface and changes of periprosthetic bone volume. RESULTS: Treatment groups receiving 5 microg and 25 microg ibandronate showed significant improvement of osseointegrated implant surface compared to the control group. Enhancement of periprosthetic bone volume was revealed in all treatment groups but only application of 25 microg ibandronate was significantly improved compared to the control group. CONCLUSION: A minor dose of 1 microg ibandronate is not effective to improve osseointegration. A high dosed bisphosphonate treatment with 5 microg or 25 microg ibandronate is potent to improve osseointegrated implant surface significantly compared to an untreated control in both uncoated and hydroxyapatite-coated titanium implants and to enhance periprosthetic bone volume. By that, improved secondary stability and prolonged survival time of cementless metal implants can be expected. PMID- 16380897 TI - [Effects of high-energy shock waves on the structure of the immature epiphysis--a histomorphological study]. AB - AIM: Extracorporeal shock wave treatment of infantile skeleton diseases is theoretical reasonable. The aim of this study was to investigate if extracorporeal shock wave therapy (ESWT) may cause damage to the epiphysis within the energy range of human application doses. METHOD: 18 rabbits were treated with 800 extracorporeal shock waves (Philips lithotripter LDM-E, energy density 0.32 mJ/mm(2)) at the proximal tibia epiphysis. At 72 hours, 14 days and 4 weeks each after shock wave application 6 rabbits were sacrificed. RESULTS: Histological examination showed no damage to the epiphysis in all treated subjects. CONCLUSION: This experimental study demonstrates that, under the conditions used in human shock wave therapy, no damage to the rabbit epiphysis occurs. PMID- 16380898 TI - [Case report of Ito-syndrome associated with congenital hemihypertrophy from the orthopaedic point of view]. AB - We report on the orthopaedic treatment of a patient with the very rare Ito syndrome and congenital hemihypertrophy. The leading symptom is the lamellar depigmentation of the skin for which it is synonymously called incontinantia pigmenti acromians. Further anomalies are found in the central nervous system, as well as the ocular and the musculoskeletal systems. The treatment of the hemihypertrophy and the coexistent dysplasia of the hip with a combination of intertrochanteric shortening osteotomy and a triple osteotomy are specified and further methods are discussed. PMID- 16380899 TI - [Biocompatibility testing of different sterilised or disinfected allogenous bone grafts in comparison to the gold standard of autologous bone grafts--an "in vitro" analysis of immunomodulation]. AB - INTRODUCTION: Repair of large skeletal defects using bone allografts has become a routine procedure in orthopaedic and trauma surgery. Different procedures of sterilisation (82.5 degrees C disinfection; 121 degrees C autoclaving; PES; Tutoplast; 25 kGy gamma irradiation) are available to inactivate bacteria and fungi, including their spores, as well as viruses in human bone allografts. The efficiency of these procedures has been proven. However, the effects on the cellular response are rarely investigated. This present in vitro study investigates the immunological answer of human bone marrow cells to human allogenous and autologous bone platelets which were sterilised by different methods. MATERIALS AND METHODS: Human bone marrow cells and the bone platelets were harvested from patients undergoing a total hip replacement. All patients provided informed consent. Human bone platelets, 10 mm in diameter, 3 mm in height, were produced from femoral heads which were removed within the scope of total hip replacements. They were sterilised by different procedures or were disinfected (gamma radiotherapy, PES/ethanol treatment, Tutoplast procedure, 121 degrees C autoclaving, > 82.5 degrees C thermodisinfection). In addition, an autologous in vitro bone donation was simulated and compared with the allogenous bone grafts. Endobon was evaluated as a bovine hydroxyapatite ceramic. As control a human bone marrow cell culture without bone platelets was used. Over a period of four weeks the changes of the immunogenic cell populations were analysed in vitro (FACS analysis). Light and scanning microscopy were done to reveal morphological differences. As a vitality test the trypan-blue staining was performed. RESULTS: Light and scanning microscopy demonstrated large differences between the various sterilisation and disinfection methods. After 4 weeks the autologous bone platelets were completely covered with homogenously distributed human osteoblast like cells. The heat-sterilised/disinfected transplants demonstrated similar effects compared to the autologous bone grafts while the irradiated bone platelets demonstrated less cell coverage. 2/3 of the cells were vital on average after four weeks, with the exception of the irradiated bone platelets. The FACS analysis revealed in comparison to the control group provable differences in the immunological answer for the autologous bone donation as well as for the differently sterilised or disinfected allogenous bone grafts. The heat sterilisation or, respectively, disinfection methods compared to the autologous bone donation demonstrated almost similar in vitro effects. By far the worst results, characterised by an excessively increased portion of cytotoxic T-cells and a decreased amount of viable cells, were seen in the 25 kGy gamma irradiation samples. CONCLUSIONS: The results demonstrate the influence of the different sterilisation and disinfection procedures on the differentiation of human marrow cells (host). Similar in vitro effects were seen for the autologous and heat treated bone platelets. The treatment of allogenous bone grafts with PES/ethanol and the Tutoplast procedures showed, just as Endobon, only low differences in comparison with the control cultures. The worse results in the case of the irradiated bone platelets may be explained by the production of free radicals which led to an excessive cell death. PMID- 16380900 TI - [Cell differentiation of a human bone marrow cell culture under the influence of UHMW-PE debris]. AB - AIM: The purpose of the present study was to evaluate the influence of ultra-high molecular-weight polyethylene (UHMW-PE), which is the major constituent of the material debris formed as a result of orthopaedic implant wear, on the cellular differentiation in a modified in vitro model. METHODS: UHMW-PE particles (O < or = 7.5 microm) were suspended in soluble collagen type I and subsequently solidified in different concentrations (10(5), 10(6) and 10(7) particles per well) on the bottom of the wells. Human bone marrow cells in a concentration of 3 x 10(6) cells per well were seeded on the collagen-particle substrate and maintained for up to 72 h. The response of the cells to the particles was examined by light microscopy, scanning electron microscopy and FACS analysis compared to cells on control collagen surfaces without any particles. RESULTS: Light and scanning microscopic evaluation revealed that the UHMW-PE particles, which had built large conglomerates (O 7.5 microm), were mainly surrounded by the cells and less phagocytosed. The results of the FACS analysis revealed significant differences in CD3/CD4 positive, CD14 positive and CD19 positive cells (p < 0.05). A significant elevation of CD3/CD4 positive and CD14 positive cells (p < 0.05) was observed after the period of culture (72 h) whereas a significant decrease could be detected in the case of CD19 positive cells. CONCLUSION: The results demonstrate that the particle-induced response by UHMW-PE limits itself not only to the particle macrophage contact but influences also the differentiation of the bone marrow. Moreover, the results confirm that the present method is useful to evaluate the in vitro effects of UHMW-PE wear particles with direct particle cell contact. Although the particles built large conglomerates, it could be shown that a change of the immune-competent cells also occurred. PMID- 16380901 TI - [Refined adenoviral transduction for controlled gene transfer into human adult mesenchymal stem cells]. AB - AIM: Adenoviral gene transfer remains a powerful tool for basic research purposes. We hypothesize that adenoviral transduction of human mesenchymal stem cells (hMSC) in vitro can be improved by refined use of experimental parameters. METHODS: hMSCs were transduced by adenoviral vectors encoding Luciferase or BMP-2 at a selection of multiplicities of infection (MOI) and exposure times. Transgene production and total protein content were measured. To determine practical relevance, expression of the bone marker genes Runx2 and Type I collagen was analyzed by quantitative PCR. As a phenotypic marker alkaline phosphatase was assessed. ANOVA and post hoc statistical analyses were used to determine differences among data (p < 0.05). RESULTS: Prolonged exposure led to a decrease in transgene production and total protein content. Increasing MOI at exposure of up to 4 hours resulted in a higher production of the transgene. Transfer of the hBMP-2 gene promoted an enhanced lineage progression to the osteoblast phenotype indicating biological activity. CONCLUSION: Time of exposure is of major importance for toxicity in vitro and should not exceed 4 hours for hMSC. While increase in exposure time leads to cell death, surviving cells, up to a certain limit, seem to compensate by increasing production of the transgene indicating that transduction efficiency cannot be positively measured in a binary yes-or-no scheme. PMID- 16380902 TI - [Osteoblastic differentiation of human adult mesenchymal stem cells after through gene transfer of BMP-2 in the absence of dexamethasone]. AB - AIM: Mesenchymal stem cells (MSC) of various species appear to require different cues to differentiate towards the osteoblastic lineage. For MSC of human origin, recombinant hBMP-2 is reported to be not sufficient but dexamethasone seems to be essential. The aim of this study was to analyse changes in genotype and phenotype of hMSC after adenoviral transfer of the BMP-2 gene in the absence of dexamethasone. METHODS: We employed hMSC and analysed changes in expression of the Runx2, Osterix and type I collagen gene by quantitative PCR after adenoviral transfer of the human BMP-2 gene in the absence of dexamethasone. As a phenotypic marker alkaline phosphatase activity was assessed. ANOVA and post hoc statistical analyses were used to determine differences among data (p < 0.05). RESULTS: Transfer of the hBMP-2 gene and consecutive production of transgenic BMP-2 up regulated bone marker gene expression and increased alkaline phosphatase activity and thus promoted an enhanced lineage progression to the osteoblast phenotype without the addition of dexamethasone. CONCLUSION: These findings are noteworthy in the light of a possible superiority of endogenous transgenic proteins compared to exogenous recombinant proteins. PMID- 16380903 TI - [Isolated bilateral Sternocostoclavicular Hyperostosis (SCCH) with a pathological clavicular fracture--a 10-year follow-up]. AB - The presented case demonstrates the clinical and radiological course of a woman suffering from bilateral sternocostoclavicular hyperostosis (SCCH) from early changes to a spontaneous clavicula fracture after 10 years. SCCH is characterised by a chronic recurrent painful swelling of the sternoclavicular region due to an aseptic inflammation and hyperostosis of the clavicula, sternum, upper ribs and the adjacent soft tissues. Frequently the picture is accompanied by cutaneous and other skeletal symptoms. The disease can be part of the SAPHO (synovitis, acne, pustulosis, hyperostosis, osteitis) syndrome. The chronic process is represented by the typical "bull horn sign" in a bone scan. NSAIDS should represent the first line treatment. PMID- 16380904 TI - A homozygous missense mutation in TGM5 abolishes epidermal transglutaminase 5 activity and causes acral peeling skin syndrome. AB - Peeling skin syndrome is an autosomal recessive genodermatosis characterized by the shedding of the outer epidermis. In the acral form, the dorsa of the hands and feet are predominantly affected. Ultrastructural analysis has revealed tissue separation at the junction between the granular cells and the stratum corneum in the outer epidermis. Genomewide linkage analysis in a consanguineous Dutch kindred mapped the gene to 15q15.2 in the interval between markers D15S1040 and D15S1016. Two homozygous missense mutations, T109M and G113C, were found in TGM5, which encodes transglutaminase 5 (TG5), in all affected persons in two unrelated families. The mutation was present on the same haplotype in both kindreds, indicating a probable ancestral mutation. TG5 is strongly expressed in the epidermal granular cells, where it cross-links a variety of structural proteins in the terminal differentiation of the epidermis to form the cornified cell envelope. An established, in vitro, biochemical cross-linking assay revealed that, although T109M is not pathogenic, G113C completely abolishes TG5 activity. Three-dimensional modeling of TG5 showed that G113C lies close to the catalytic domain, and, furthermore, that this glycine residue is conserved in all known transglutaminases, which is consistent with pathogenicity. Other families with more-widespread peeling skin phenotypes lacked TGM5 mutations. This study identifies the first causative gene in this heterogeneous group of skin disorders and demonstrates that the protein cross-linking function performed by TG5 is vital for maintaining cell-cell adhesion between the outermost layers of the epidermis. PMID- 16380906 TI - Genomewide high-density SNP linkage analysis of 236 Japanese families supports the existence of schizophrenia susceptibility loci on chromosomes 1p, 14q, and 20p. AB - The Japanese Schizophrenia Sib-Pair Linkage Group (JSSLG) is a multisite collaborative study group that was organized to create a national resource for affected sib pair (ASP) studies of schizophrenia in Japan. We used a high-density single-nucleotide-polymorphism (SNP) genotyping assay, the Illumina BeadArray linkage mapping panel (version 4) comprising 5,861 SNPs, to perform a genomewide linkage analysis of JSSLG samples comprising 236 Japanese families with 268 nonindependent ASPs with schizophrenia. All subjects were Japanese. Among these families, 122 families comprised the same subjects analyzed with short tandem repeat markers. All the probands and their siblings, with the exception of seven siblings with schizoaffective disorder, had schizophrenia. After excluding SNPs with high linkage disequilibrium, we found significant evidence of linkage of schizophrenia to chromosome 1p21.2-1p13.2 (LOD=3.39) and suggestive evidence of linkage to 14q11.2 (LOD=2.87), 14q11.2-q13.2 (LOD=2.33), and 20p12.1-p11.2 (LOD=2.33). Although linkage to these regions has received little attention, these regions are included in or partially overlap the 10 regions reported by Lewis et al. that passed the two aggregate criteria of a meta-analysis. Results of the present study--which, to our knowledge, is the first genomewide analysis of schizophrenia in ASPs of a single Asian ethnicity that is comparable to the analyses done of ASPs of European descent--indicate the existence of schizophrenia susceptibility loci that are common to different ethnic groups but that likely have different ethnicity-specific effects. PMID- 16380905 TI - Bipolar I disorder and schizophrenia: a 440-single-nucleotide polymorphism screen of 64 candidate genes among Ashkenazi Jewish case-parent trios. AB - Bipolar, schizophrenia, and schizoaffective disorders are common, highly heritable psychiatric disorders, for which familial coaggregation, as well as epidemiological and genetic evidence, suggests overlapping etiologies. No definitive susceptibility genes have yet been identified for any of these disorders. Genetic heterogeneity, combined with phenotypic imprecision and poor marker coverage, has contributed to the difficulty in defining risk variants. We focused on families of Ashkenazi Jewish descent, to reduce genetic heterogeneity, and, as a precursor to genomewide association studies, we undertook a single nucleotide polymorphism (SNP) genotyping screen of 64 candidate genes (440 SNPs) chosen on the basis of previous linkage or of association and/or biological relevance. We genotyped an average of 6.9 SNPs per gene, with an average density of 1 SNP per 11.9 kb in 323 bipolar I disorder and 274 schizophrenia or schizoaffective Ashkenazi case-parent trios. Using single-SNP and haplotype-based transmission/disequilibrium tests, we ranked genes on the basis of strength of association (P<.01). Six genes (DAO, GRM3, GRM4, GRIN2B, IL2RB, and TUBA8) met this criterion for bipolar I disorder; only DAO has been previously associated with bipolar disorder. Six genes (RGS4, SCA1, GRM4, DPYSL2, NOS1, and GRID1) met this criterion for schizophrenia or schizoaffective disorder; five replicate previous associations, and one, GRID1, shows a novel association with schizophrenia. In addition, six genes (DPYSL2, DTNBP1, G30/G72, GRID1, GRM4, and NOS1) showed overlapping suggestive evidence of association in both disorders. These results may help to prioritize candidate genes for future study from among the many suspected/proposed for schizophrenia and bipolar disorders. They provide further support for shared genetic susceptibility between these two disorders that involve glutamate-signaling pathways. PMID- 16380908 TI - Preferential transmission of paternal alleles at risk genes in attention deficit/hyperactivity disorder. AB - Family, twin, and adoption studies have demonstrated a significant genetic contribution to the etiology of attention-deficit/hyperactivity disorder (ADHD). Pharmacological, neuroimaging, and animal-model findings suggest imbalances in monoaminergic (dopaminergic, serotonergic, and noradrenergic) neurotransmission in ADHD. We have examined monoaminergic candidate genes for possible genetic association with ADHD in the Irish population, focusing particularly on genes of the dopaminergic and serotonergic systems. We have observed that several of these genes are associated with ADHD, including DAT1, DBH, DRD4, DRD5, and 5HT1B. Here, we present what appears to be a systematic overtransmission of paternal alleles at candidate genes associated with ADHD. For the nine genes included in the analysis, the overall odds ratio for paternal transmission was 2, compared with 1.3 for maternal transmission (paternal vs. maternal chi 2=9.6; P=.0019). Transmission to females, from either parent, was significantly stronger than to males. Possible reasons for this preferential transmission include imprinting and ascertainment bias, although results of further analyses show that the latter is unlikely. PMID- 16380907 TI - GJB2 mutations and degree of hearing loss: a multicenter study. AB - Hearing impairment (HI) affects 1 in 650 newborns, which makes it the most common congenital sensory impairment. Despite extraordinary genetic heterogeneity, mutations in one gene, GJB2, which encodes the connexin 26 protein and is involved in inner ear homeostasis, are found in up to 50% of patients with autosomal recessive nonsyndromic hearing loss. Because of the high frequency of GJB2 mutations, mutation analysis of this gene is widely available as a diagnostic test. In this study, we assessed the association between genotype and degree of hearing loss in persons with HI and biallelic GJB2 mutations. We performed cross-sectional analyses of GJB2 genotype and audiometric data from 1,531 persons, from 16 different countries, with autosomal recessive, mild-to profound nonsyndromic HI. The median age of all participants was 8 years; 90% of persons were within the age range of 0-26 years. Of the 83 different mutations identified, 47 were classified as nontruncating, and 36 as truncating. A total of 153 different genotypes were found, of which 56 were homozygous truncating (T/T), 30 were homozygous nontruncating (NT/NT), and 67 were compound heterozygous truncating/nontruncating (T/NT). The degree of HI associated with biallelic truncating mutations was significantly more severe than the HI associated with biallelic nontruncating mutations (P<.0001). The HI of 48 different genotypes was less severe than that of 35delG homozygotes. Several common mutations (M34T, V37I, and L90P) were associated with mild-to-moderate HI (median 25-40 dB). Two genotypes--35delG/R143W (median 105 dB) and 35delG/dela(GJB6-D13S1830) (median 108 dB)--had significantly more-severe HI than that of 35delG homozygotes. PMID- 16380909 TI - Heterogeneous duplications in patients with Pelizaeus-Merzbacher disease suggest a mechanism of coupled homologous and nonhomologous recombination. AB - We describe genomic structures of 59 X-chromosome segmental duplications that include the proteolipid protein 1 gene (PLP1) in patients with Pelizaeus Merzbacher disease. We provide the first report of 13 junction sequences, which gives insight into underlying mechanisms. Although proximal breakpoints were highly variable, distal breakpoints tended to cluster around low-copy repeats (LCRs) (50% of distal breakpoints), and each duplication event appeared to be unique (100 kb to 4.6 Mb in size). Sequence analysis of the junctions revealed no large homologous regions between proximal and distal breakpoints. Most junctions had microhomology of 1-6 bases, and one had a 2-base insertion. Boundaries between single-copy and duplicated DNA were identical to the reference genomic sequence in all patients investigated. Taken together, these data suggest that the tandem duplications are formed by a coupled homologous and nonhomologous recombination mechanism. We suggest repair of a double-stranded break (DSB) by one-sided homologous strand invasion of a sister chromatid, followed by DNA synthesis and nonhomologous end joining with the other end of the break. This is in contrast to other genomic disorders that have recurrent rearrangements formed by nonallelic homologous recombination between LCRs. Interspersed repetitive elements (Alu elements, long interspersed nuclear elements, and long terminal repeats) were found at 18 of the 26 breakpoint sequences studied. No specific motif that may predispose to DSBs was revealed, but single or alternating tracts of purines and pyrimidines that may cause secondary structures were common. Analysis of the 2-Mb region susceptible to duplications identified proximal specific repeats and distal LCRs in addition to the previously reported ones, suggesting that the unique genomic architecture may have a role in nonrecurrent rearrangements by promoting instability. PMID- 16380910 TI - The effect of single-nucleotide polymorphism marker selection on patterns of haplotype blocks and haplotype frequency estimates. AB - The definition of haplotype blocks of single-nucleotide polymorphisms (SNPs) has been proposed so that the haplotypes can be used as markers in association studies and to efficiently describe human genetic variation. The International Haplotype Map (HapMap) project to construct a comprehensive catalog of haplotypic variation in humans is underway. However, a number of factors have already been shown to influence the definition of blocks, including the population studied and the sample SNP density. Here, we examine the effect that marker selection has on the definition of blocks and the pattern of haplotypes by using comparable but complementary SNP sets and a number of block definition methods in various genomic regions and populations that were provided by the Encyclopedia of DNA Elements (ENCODE) project. We find that the chosen SNP set has a profound effect on the block-covered sequence and block borders, even at high marker densities. Our results question the very concept of discrete haplotype blocks and the possibility of generalizing block findings from the HapMap project. We comparatively apply the block-free tagging-SNP approach and discuss both the haplotype approach and the tagging-SNP approach as means to efficiently catalog genetic variation. PMID- 16380911 TI - Discriminating power of localized three-dimensional facial morphology. AB - Many genetic syndromes involve a facial gestalt that suggests a preliminary diagnosis to an experienced clinical geneticist even before a clinical examination and genotyping are undertaken. Previously, using visualization and pattern recognition, we showed that dense surface models (DSMs) of full face shape characterize facial dysmorphology in Noonan and in 22q11 deletion syndromes. In this much larger study of 696 individuals, we extend the use of DSMs of the full face to establish accurate discrimination between controls and individuals with Williams, Smith-Magenis, 22q11 deletion, or Noonan syndromes and between individuals with different syndromes in these groups. However, the full power of the DSM approach is demonstrated by the comparable discriminating abilities of localized facial features, such as periorbital, perinasal, and perioral patches, and the correlation of DSM-based predictions and molecular findings. This study demonstrates the potential of face shape models to assist clinical training through visualization, to support clinical diagnosis of affected individuals through pattern recognition, and to enable the objective comparison of individuals sharing other phenotypic or genotypic properties. PMID- 16380912 TI - A genomewide linkage study of 1,933 families affected by premature coronary artery disease: The British Heart Foundation (BHF) Family Heart Study. AB - Coronary artery disease (CAD) and its most important complication, myocardial infarction (MI), are the leading cause of premature death in the Western world. CAD has a substantial genetic basis, especially when it occurs early. We investigated the genetic determinants of premature CAD by performing a genomewide linkage analysis of 4,175 affected subjects from 1,933 families recruited throughout the United Kingdom. Each family had at least two available siblings with CAD, with validated onset before age 66 years. Linkage analysis was performed using 416 microsatellite markers. We observed suggestive linkage, for both CAD and MI, to a region on chromosome 2. For CAD, a LOD score of 1.86 was observed at marker D2S2271, which, in an ordered subset analysis, increased to 2.70 in families (n=1,698) with a minimum age at diagnosis of 56 years or younger. For MI, an overlapping peak with a LOD score of 1.15 was observed at marker D2S2216, which increased to 2.1 in families (n=801) with a minimum age at diagnosis of 59 years or younger. Exclusion mapping showed that 100% of the autosomal genome could be excluded for locus-specific sibling relative risks of 1.5 and 1.6 for CAD and MI, respectively. The region identified on chromosome 2 overlaps linked regions observed in two other smaller genome scans for CAD. Together, these findings strongly suggest that there is a locus on chromosome 2 that influences coronary atherosclerosis risk. The exclusion of a common locus that increases risk of CAD to siblings by >50% has important implications for strategies for further defining the genetic basis of CAD. PMID- 16380913 TI - Comparative genomics and gene expression analysis identifies BBS9, a new Bardet Biedl syndrome gene. AB - Bardet-Biedl syndrome (BBS) is an autosomal recessive, genetically heterogeneous, pleiotropic human disorder characterized by obesity, retinopathy, polydactyly, renal and cardiac malformations, learning disabilities, and hypogenitalism. Eight BBS genes representing all known mapped loci have been identified. Mutation analysis of the known BBS genes in BBS patients indicate that additional BBS genes exist and/or that unidentified mutations exist in the known genes. To identify new BBS genes, we performed homozygosity mapping of small, consanguineous BBS pedigrees, using moderately dense SNP arrays. A bioinformatics approach combining comparative genomic analysis and gene expression studies of a BBS-knockout mouse model was used to prioritize BBS candidate genes within the newly identified loci for mutation screening. By use of this strategy, parathyroid hormone-responsive gene B1 (B1) was found to be a novel BBS gene (BBS9), supported by the identification of homozygous mutations in BBS patients. The identification of BBS9 illustrates the power of using a combination of comparative genomic analysis, gene expression studies, and homozygosity mapping with SNP arrays in small, consanguineous families for the identification of rare autosomal recessive disorders. We also demonstrate that small, consanguineous families are useful in identifying intragenic deletions. This type of mutation is likely to be underreported because of the difficulty of deletion detection in the heterozygous state by the mutation screening methods that are used in many studies. PMID- 16380914 TI - The Y deletion gr/gr and susceptibility to testicular germ cell tumor. AB - Testicular germ cell tumor (TGCT) is the most common cancer in young men. Despite a considerable familial component to TGCT risk, no genetic change that confers increased risk has been substantiated to date. The human Y chromosome carries a number of genes specifically involved in male germ cell development, and deletion of the AZFc region at Yq11 is the most common known genetic cause of infertility. Recently, a 1.6-Mb deletion of the Y chromosome that removes part of the AZFc region--known as the "gr/gr" deletion--has been associated with infertility. In epidemiological studies, male infertility has shown an association with TGCT that is out of proportion with what can be explained by tumor effects. Thus, we hypothesized that the gr/gr deletion may be associated with TGCT. Using logistic modeling, we analyzed this deletion in a large series of TGCT cases with and without a family history of TGCT. The gr/gr deletion was present in 3.0% (13/431) of TGCT cases with a family history, 2% (28/1,376) of TGCT cases without a family history, and 1.3% (33/2,599) of unaffected males. Presence of the gr/gr deletion was associated with a twofold increased risk of TGCT (adjusted odds ratio [aOR] 2.1; 95% confidence interval [CI] 1.3-3.6; P = .005) and a threefold increased risk of TGCT among patients with a positive family history (aOR 3.2; 95% CI 1.5 6.7; P = .0027). The gr/gr deletion was more strongly associated with seminoma (aOR 3.0; 95% CI 1.6-5.4; P = .0004) than with nonseminoma TGCT (aOR 1.5; 95% CI 0.72-3.0; P = .29). These data indicate that the Y microdeletion gr/gr is a rare, low-penetrance allele that confers susceptibility to TGCT. PMID- 16380916 TI - The role of mannosylated enzyme and the mannose receptor in enzyme replacement therapy. AB - Lysosomal acid lipase (LAL) is the critical enzyme for the hydrolysis of triglycerides (TGs) and cholesteryl esters (CEs) in lysosomes. LAL defects cause Wolman disease (WD) and CE storage disease (CESD). An LAL null (lal-/-) mouse model closely mimics human WD/CESD, with hepatocellular, Kupffer cell and other macrophage, and adrenal cortical storage of CEs and TGs. The effect on the cellular targeting of high-mannose and complex oligosaccharide-type oligosaccharide chains was tested with human LAL expressed in Pichia pastoris (phLAL) and CHO cells (chLAL), respectively. Only chLAL was internalized by cultured fibroblasts, whereas both chLAL and phLAL were taken up by macrophage mannose receptor (MMR)-positive J774E cells. After intraperitoneal injection into lal-/- mice, phLAL and chLAL distributed to macrophages and macrophage-derived cells of various organs. chLAL was also detected in hepatocytes. Ten injections of either enzyme over 30 d into 2- and 2.5-mo-old lal-/- mice produced normalization of hepatic color, decreased liver weight (50%-58%), and diminished hepatic cholesterol and TG storage. Lipid accumulations in macrophages were diminished with either enzyme. Only chLAL cleared lipids in hepatocytes. Mice double homozygous for the LAL and MMR deficiences (lal-/-;MMR-/-) showed phLAL uptake into Kupffer cells and hepatocytes, reversal of macrophage histopathology and lipid storage in all tissues, and clearance of hepatocytes. These results implicate MMR-independent and mannose 6-phosphate receptor-independent pathways in phLAL uptake and delivery to lysosomes in vivo. In addition, these studies show specific cellular targeting and physiologic effects of differentially oligosaccharide-modified human LALs mediated by MMR and that lysosomal targeting of mannose-terminated glycoproteins occurs and storage can be eliminated effectively without MMR. PMID- 16380915 TI - Replication of putative candidate-gene associations with rheumatoid arthritis in >4,000 samples from North America and Sweden: association of susceptibility with PTPN22, CTLA4, and PADI4. AB - Candidate-gene association studies in rheumatoid arthritis (RA) have lead to encouraging yet apparently inconsistent results. One explanation for the inconsistency is insufficient power to detect modest effects in the context of a low prior probability of a true effect. To overcome this limitation, we selected alleles with an increased probability of a disease association, on the basis of a review of the literature on RA and other autoimmune diseases, and tested them for association with RA susceptibility in a sample collection powered to detect modest genetic effects. We tested 17 alleles from 14 genes in 2,370 RA cases and 1,757 controls from the North American Rheumatoid Arthritis Consortium (NARAC) and the Swedish Epidemiological Investigation of Rheumatoid Arthritis (EIRA) collections. We found strong evidence of an association of PTPN22 with the development of anti-citrulline antibody-positive RA (odds ratio [OR] 1.49; P=.00002), using previously untested EIRA samples. We provide support for an association of CTLA4 (CT60 allele, OR 1.23; P=.001) and PADI4 (PADI4_94, OR 1.24; P=.001) with the development of RA, but only in the NARAC cohort. The CTLA4 association is stronger in patients with RA from both cohorts who are seropositive for anti-citrulline antibodies (P=.0006). Exploration of our data set with clinically relevant subsets of RA reveals that PTPN22 is associated with an earlier age at disease onset (P=.004) and that PTPN22 has a stronger effect in males than in females (P=.03). A meta-analysis failed to demonstrate an association of the remaining alleles with RA susceptibility, suggesting that the previously published associations may represent false-positive results. Given the strong statistical power to replicate a true-positive association in this study, our results provide support for PTPN22, CTLA4, and PADI4 as RA susceptibility genes and demonstrate novel associations with clinically relevant subsets of RA. PMID- 16380917 TI - Robust estimation of experimentwise P values applied to a genome scan of multiple asthma traits identifies a new region of significant linkage on chromosome 20q13. AB - Over 30 genomic regions show linkage to asthma traits. Six asthma genes have been cloned, but the putative loci in many linked regions have not been identified. To search for asthma susceptibility loci, we performed genomewide univariate linkage analyses of seven asthma traits, using 202 Australian families ascertained through a twin proband. House-dust mite sensitivity (Dpter) exceeded the empirical threshold for significant linkage at 102 cM on chromosome 20q13, near marker D20S173 (empirical pointwise P = .00001 and genomewide P = .005, both uncorrected for multiple-trait testing). Atopy, bronchial hyperresponsiveness (BHR), and forced expiratory volume in 1 s (FEV1) were also linked to this region. In addition, 16 regions were linked to at least one trait at the suggestive level, including 12q24, which has consistently shown linkage to asthma traits in other studies. Some regions were expected to be false-positives arising from multiple-trait testing. To address this, we developed a new approach to estimate genomewide significance that accounts for multiple-trait testing and for correlation between traits and that does not require a Bonferroni correction. With this approach, Dpter remained significantly linked to 20q13 (empirical genomewide P = .042), and airway obstruction remained linked to 12q24 at the suggestive level. Finally, we extended this method to show that the linkage of Dpter, atopy, BHR, FEV1, asthma, and airway obstruction to chromosome 20q13 is unlikely to be due to chance and may result from a quantitative trait locus in this region that affects several of these traits. PMID- 16380918 TI - Identification of an X-chromosomal locus and haplotype modulating the phenotype of a mitochondrial DNA disorder. AB - Mitochondrial DNA (mtDNA) mutations are a major cause of human disease. A large number of different molecular defects ultimately compromise oxidative phosphorylation, but it is not clear why the same biochemical defect can cause diverse clinical phenotypes. There is emerging evidence that nuclear genes modulate the phenotype of primary mtDNA disorders. Here, we define an X chromosomal haplotype that interacts with specific MTND mutations to cause visual failure in the most common mtDNA disease, Leber hereditary optic neuropathy. This effect is independent of the mtDNA genetic background and explains the variable penetrance and sex bias that characterizes this disorder. PMID- 16380919 TI - NF1 gene mutations represent the major molecular event underlying neurofibromatosis-Noonan syndrome. AB - Neurofibromatosis type 1 (NF1) demonstrates phenotypic overlap with Noonan syndrome (NS) in some patients, which results in the so-called neurofibromatosis Noonan syndrome (NFNS). From a genetic point of view, NFNS is a poorly understood condition, and controversy remains as to whether it represents a variable manifestation of either NF1 or NS or is a distinct clinical entity. To answer this question, we screened a cohort with clinically well-characterized NFNS for mutations in the entire coding sequence of the NF1 and PTPN11 genes. Heterozygous NF1 defects were identified in 16 of the 17 unrelated subjects included in the study, which provides evidence that mutations in NF1 represent the major molecular event underlying this condition. Lesions included nonsense mutations, out-of-frame deletions, missense changes, small inframe deletions, and one large multiexon deletion. Remarkably, a high prevalence of inframe defects affecting exons 24 and 25, which encode a portion of the GAP-related domain of the protein, was observed. On the other hand, no defect in PTPN11 was observed, and no lesion affecting exons 11-27 of the NF1 gene was identified in 100 PTPN11 mutation negative subjects with NS, which provides further evidence that NFNS and NS are genetically distinct disorders. These results support the view that NFNS represents a variant of NF1 and is caused by mutations of the NF1 gene, some of which have been demonstrated to cause classic NF1 in other individuals. PMID- 16380920 TI - Genomewide scan and fine-mapping linkage studies in four European samples with bipolar affective disorder suggest a new susceptibility locus on chromosome 1p35 p36 and provides further evidence of loci on chromosome 4q31 and 6q24. AB - We present the findings of a large linkage study of bipolar affective disorder (BPAD) that involved genomewide analysis of 52 families (448 genotyped individuals) of Spanish, Romany, and Bulgarian descent and further fine mapping of the 1p34-p36, 4q28-q31, and 6q15-q24 regions. An additional sample of 56 German families (280 individuals) was included for this fine-mapping step. The highest nonparametric linkage scores obtained in the fine mapping were 5.49 for 4q31 and 4.87 for 6q24 in the Romany families and 3.97 for 1p35-p36 in the Spanish sample. MOD-score (LOD scores maximized over genetic model parameters) analysis provided significant evidence of linkage to 4q31 and at least borderline significance for the 1p and 6q regions. On the basis of these results and previous positive research findings, 4q31 and 6q24 should now be considered confirmed BPAD susceptibility loci, and 1p35-p36 is proposed as a new putative locus that requires confirmation in replication studies. PMID- 16380921 TI - Recent spread of a Y-chromosomal lineage in northern China and Mongolia. AB - We have identified a Y-chromosomal lineage that is unusually frequent in northeastern China and Mongolia, in which a haplotype cluster defined by 15 Y short tandem repeats was carried by approximately 3.3% of the males sampled from East Asia. The most recent common ancestor of this lineage lived 590 +/- 340 years ago (mean +/- SD), and it was detected in Mongolians and six Chinese minority populations. We suggest that the lineage was spread by Qing Dynasty (1644-1912) nobility, who were a privileged elite sharing patrilineal descent from Giocangga (died 1582), the grandfather of Manchu leader Nurhaci, and whose documented members formed approximately 0.4% of the minority population by the end of the dynasty. PMID- 16380923 TI - Ontogeny of performance in vertebrates. AB - When competing for food or other resources, or when confronted with predators, young animals may be at a disadvantage relative to adults because of their smaller size. Additionally, the ongoing differentiation and growth of tissues and the development of sensory-motor integration during early ontogeny may constrain performance. Because ectothermic vertebrates show different growth regimes and energetic requirements when compared to endothermic vertebrates, differences in the ontogenetic trajectories of performance traits in these two groups might be expected. However, both groups of vertebrates show similar patterns of changes in performance with ontogeny. Evidence for compensation, resulting in relatively high levels of performance in juveniles relative to adults, appears common for traits related to locomotor and defensive behaviors. However, there is little evidence for compensation in traits associated with feeding and foraging. We suggest that this difference may be due to different selective regimes operating on locomotor versus feeding traits. As a result, relatively high levels of locomotor performance in juveniles and relatively high levels of feeding performance in adults are observed across a wide range of vertebrate groups. PMID- 16380922 TI - Inactivating mutations in ESCO2 cause SC phocomelia and Roberts syndrome: no phenotype-genotype correlation. AB - The rare, autosomal recessive Roberts syndrome (RBS) is characterized by tetraphocomelia, profound growth deficiency of prenatal onset, craniofacial anomalies, microcephaly, and mental deficiency. SC phocomelia (SC) has a milder phenotype, with a lesser degree of limb reduction and with survival to adulthood. Since heterochromatin repulsion (HR) is characteristic for both disorders and is not complemented in somatic-cell hybrids, it has been hypothesized that the disorders are allelic. Recently, mutations in ESCO2 (establishment of cohesion 1 homolog 2) on 8p21.1 have been reported in RBS. To determine whether ESCO2 mutations are also responsible for SC, we studied three families with SC and two families in which variable degrees of limb and craniofacial abnormalities, detected by fetal ultrasound, led to pregnancy terminations. All cases were positive for HR. We identified seven novel mutations in exons 3-8 of ESCO2. In two families, affected individuals were homozygous--for a 5-nucleotide deletion in one family and a splice-site mutation in the other. In three nonconsanguineous families, probands were compound heterozygous for a single-nucleotide insertion or deletion, a nonsense mutation, or a splice-site mutation. Abnormal splice products were characterized at the RNA level. Since only protein-truncating mutations were identified, regardless of clinical severity, we conclude that genotype does not predict phenotype. Having established that RBS and SC are caused by mutations in the same gene, we delineated the clinical phenotype of the tetraphocomelia spectrum that is associated with HR and ESCO2 mutations and differentiated it from other types of phocomelia that are negative for HR. PMID- 16380924 TI - Development of the escape response in teleost fishes: do ontogenetic changes enable improved performance? AB - Teleost fishes typically first encounter the environment as free-swimming embryos or larvae. Larvae are morphologically distinct from adults, and major anatomical structures are unformed. Thus, larvae undergo a series of dramatic morphological changes until they reach adult morphology (but are reproductively immature) and are considered juveniles. Free-swimming embryos and larvae are able to perform a C-start, an effective escape response that is used evade predators. However, escape response performance improves during early development: as young fish grow, they swim faster (length-specific maximum velocity increases) and perform the escape more rapidly (time to complete the behavior decreases). These improvements cease when fish become juveniles, although absolute swimming velocity (m s(-1)) continues to increase. We use studies of escape behavior and ontogeny in California halibut (Paralichthys californicus), rainbow trout (Oncorhynchus mykiss), and razorback suckers (Xyrauchen texanus) to test the hypothesis that specific morphological changes improve escape performance. We suggest that formation of the caudal fin improves energy transfer to the water and therefore increases thrust production and swimming velocity. In addition, changes to the axial skeleton during the larval period produce increased axial stiffness, which in turn allows the production of a more rapid and effective escape response. Because escape performance improves as adult morphology develops, fish that enter the environment in an advanced stage of development (i.e., those with direct development) should have a greater ability to evade predators than do fish that enter the environment at an early stage of development (i.e., those with indirect development). PMID- 16380925 TI - The ontogeny of contractile performance and metabolic capacity in a high frequency muscle. AB - High-performance muscles such as the shaker muscles in the tails of western diamond-backed rattlesnakes (Crotalus atrox) are excellent systems for studying the relationship between contractile performance and metabolic capacity. We observed that shaker muscle contraction frequency increases dramatically with growth in small individuals but then declines gradually in large individuals. We tested whether metabolic capacity changed with performance, using shaker muscle contraction frequency as an indicator of performance and maximal activities of citrate synthase and lactate dehydrogenase as indicators of aerobic and anaerobic capacities, respectively. Contraction frequency increased 20-fold in 20-100-g individuals but then declined by approximately 30% in individuals approaching 1,000 g. Mass-independent aerobic capacity was positively correlated with contractile performance, whereas mass-independent anaerobic capacity was slightly but negatively correlated with performance; body mass was not correlated with performance. Rattle mass increased faster than the ability to generate force. Early in ontogeny, shaker muscle performance appears to be limited by aerobic capacity, but later performance becomes limited equally by aerobic capacity and the mechanical constraint of moving a larger mass without proportionally thicker muscles. This high-performance muscle appears to shift during ontogeny from a metabolic constraint to combined metabolic and mechanical constraints. PMID- 16380926 TI - Ontogenetic scaling of bite force in lizards and turtles. AB - Because selection on juvenile life-history stages is likely strong, disproportionately high levels of performance (e.g., sprint speed, endurance, etc.) might be expected. Whereas this phenomenon has been demonstrated with respect to locomotor performance, data for feeding are scarce. Here, we investigate the relationships among body dimensions, head dimensions, and bite force during growth in lizards and turtles. We also investigate whether ontogenetic changes in bite performance are related to changes in diet. Our analyses show that, for turtles, head dimensions generally increase with negative allometry. For lizards, heads scale as expected for geometrically growing systems. Bite force generally increased isometrically with carapace length in turtles but showed significant positive allometry relative to body dimensions in lizards. However, both lizards and turtles display positive allometric scaling of bite force relative to some measures of head size throughout ontogeny, suggesting (1) strong selection for increased relative bite performance with increasing head size and (2) intrinsic changes in the geometry and/or mass of the jaw adductors during growth. Whereas our data generally do not provide strong evidence of compensation for lower absolute levels of performance, they do show strong links among morphology, bite force, and diet during growth. PMID- 16380927 TI - Scaling of suction feeding performance in the catfish Clarias gariepinus. AB - Ontogenetic changes in the absolute dimensions of the cranial system together with changes in kinematics during prey capture can cause differences in the spatiotemporal patterns of water flow generated during suction feeding. Because the velocity of this water flow determines the force that pulls prey toward and into the mouth cavity, this can affect suction feeding performance. In this study, size-related changes in the suction-induced flow patterns are determined. To do so, a mathematical suction model is applied to video recordings of prey capturing Clarias gariepinus ranging in total length from 111 to 923 mm. Although large C. gariepinus could be expected to have increasing peak velocities of water flow compared with small individuals, the results from the hydrodynamic model show that this is not the case. Yet, when C. gariepinus becomes larger, the expansive phase is prolonged, resulting in a longer sustained flow. This flow also reaches farther in front of the mouth almost proportionally with head size. Forward dynamical simulations with spherical prey that are subjected to the calculated water flows indicate that the absolute distance from which a given prey can be sucked into the mouth as well as the maximal prey diameter increase substantially with increasing head size. Consequently, the range of potential prey that can be captured through suction feeding will become broader during growth of C. gariepinus. This appears to be reflected in the natural diet of this species, where both the size and the number of evasive prey increase with increasing predator size. PMID- 16380928 TI - In vivo bone strain and ontogenetic growth patterns in relation to life-history strategies and performance in two vertebrate taxa: goats and emu. AB - This study examined ontogenetic patterns of limb loading, bone strains, and relative changes in bone geometry to explore the relationship between in vivo mechanics and size-related changes in the limb skeleton of two vertebrate taxa. Despite maintaining similar relative limb loads during ontogeny, bone strain magnitudes in the goat radius and emu tibiotarsus generally increased. However, while the strain increases in the emu tibiotarsus were mostly insignificant, strains within the radii of adult goats were two to four times greater than in young goats. The disparity between ontogenetic strain increases in these taxa resulted from differences in ontogenetic scaling patterns of the cross-sectional bone geometry. While the cross-sectional and second moments of area scaled with negative allometry in the goat radius, these measures were not significantly different from isometry in the emu tibiotarsus. Although the juveniles of both taxa exhibited lower strains and higher safety factors than the adults, the radii of the young goats were more robust relative to the adult goats than were the tibiotarsi of the young compared with adult emu. Differences in ontogenetic growth and strain patterns in the limb bones examined likely result from different threat avoidance strategies and selection pressures in the juveniles of these two taxa. PMID- 16380929 TI - Age-specific forced polymorphism: implications of ontogenetic changes in morphology for male mating tactics. AB - Age-specific forced polymorphism is the presence of two or more distinct phenotypes (here we consider only males) that occur in separate sexually mature age groups (e.g., horns in older males but not younger males). The life-stage morph maturation hypothesis posits that all younger males that possess a particular structure can transform into older males with a different structure, most likely via the influence of hormones. The life-stage morph selection hypothesis posits that polymorphism is due to intense selection resulting in a highly nonrandom sample of younger males surviving to become older males, thus leading to different mean phenotypes in different age groups. We conducted an extensive review of literature from the past 20 years (1983-2003) for cases of age-specific forced polymorphism. Overall, we found only a few cases that fit our criteria of age-specific forced polymorphism, and we argue that most (e.g., orangutans, elephant seals) have likely arisen via the life-stage morph maturation mechanism, but we also present several examples (e.g., green anole lizards) that appear to be candidates for life-stage morph selection. However, none of the reviewed studies provided enough information (e.g., age of morphs, growth patterns of the morphological structure) to definitively invoke either of the two mechanisms. We suggest that age-specific forced polymorphism is more common than reflected in this review and that future studies should gather demographic and laboratory data that will directly compare the life-stage morph maturation and life-stage morph selection hypotheses. PMID- 16380930 TI - Effects of size, sex, and voluntary running speeds on costs of locomotion in lines of laboratory mice selectively bred for high wheel-running activity. AB - Selective breeding for over 35 generations has led to four replicate (S) lines of laboratory house mice (Mus domesticus) that run voluntarily on wheels about 170% more than four random-bred control (C) lines. We tested whether S lines have evolved higher running performance by increasing running economy (i.e., decreasing energy spent per unit of distance) as a correlated response to selection, using a recently developed method that allows for nearly continuous measurements of oxygen consumption (VO2) and running speed in freely behaving animals. We estimated slope (incremental cost of transport [COT]) and intercept for regressions of power (the dependent variable, VO2/min) on speed for 49 males and 47 females, as well as their maximum VO2 and speeds during wheel running, under conditions mimicking those that these lines face during the selection protocol. For comparison, we also measured COT and maximum aerobic capacity (VO2max) during forced exercise on a motorized treadmill. As in previous studies, the increased wheel running of S lines was mainly attributable to increased average speed, with males also showing a tendency for increased time spent running. On a whole-animal basis, combined analysis of males and females indicated that COT during voluntary wheel running was significantly lower in the S lines (one-tailed P=0.015). However, mice from S lines are significantly smaller and attain higher maximum speeds on the wheels; with either body mass or maximum speed (or both) entered as a covariate, the statistical significance of the difference in COT is lost (one-tailed P> or =0.2). Thus, both body size and behavior are key components of the reduction in COT. Several statistically significant sex differences were observed, including lower COT and higher resting metabolic rate in females. In addition, maximum voluntary running speeds were negatively correlated with COT in females but not in males. Moreover, males (but not females) from the S lines exhibited significantly higher treadmill VO2max as compared to those from C lines. The sex-specific responses to selection may in part be consequences of sex differences in body mass and running style. Our results highlight how differences in size and running speed can account for lower COT in S lines and suggest that lower COT may have coadapted in response to selection for higher running distances in these lines. PMID- 16380931 TI - Routine and active metabolic rates of migrating adult wild sockeye salmon (Oncorhynchus nerka Walbaum) in seawater and freshwater. AB - We present the first data on the differences in routine and active metabolic rates for sexually maturing migratory adult sockeye salmon (Oncorhynchus nerka) that were intercepted in the ocean and then held in either seawater or freshwater. Routine and active oxygen uptake rates (MO2) were significantly higher (27%-72%) in seawater than in freshwater at all swimming speeds except those approaching critical swimming speed. During a 45-min recovery period, the declining postexercise oxygen uptake remained 58%-73% higher in seawater than in freshwater. When fish performed a second swim test, active metabolic rates again remained 28%-81% higher for fish in seawater except at the critical swimming speed. Despite their differences in metabolic rates, fish in both seawater and freshwater could repeat the swim test and reach a similar maximum oxygen uptake and critical swimming speed as in the first swim test, even without restoring routine metabolic rate between swim tests. Thus, elevated MO2 related to either being in seawater as opposed to freshwater or not being fully recovered from previous exhaustive exercise did not present itself as a metabolic loading that limited either critical swimming performance or maximum MO2. The basis for the difference in metabolic rates of migratory sockeye salmon held in seawater and freshwater is uncertain, but it could include differences in states of nutrition, reproduction, and restlessness, as well as ionic differences. Regardless, this study elucidates some of the metabolic costs involved during the migration of adult salmon from seawater to freshwater, which may have applications for fisheries conservation and management models of energy use. PMID- 16380932 TI - Changes in the condition factor have an impact on metabolic rate and swimming performance relationships in Atlantic cod (Gadus morhua L.). AB - In the field, Atlantic cod face seasonal changes in food availability that in turn lead to changes in condition. To examine the physiological consequences of these changes in condition, we measured routine metabolic rate (RMR) to estimate standard metabolic rate (SMR), active metabolic rate (AMR), aerobic scope, critical swimming speed (Ucrit), cost of transport, sprint performance, time to exhaustion, and postexhaustion metabolic rate (EMR) for 24 Atlantic cod from the Gulf of St. Lawrence. Cod were measured at their initial condition (condition factor of 0.676+/-0.076) and after 9 wk of feeding (condition factor of 0.923+/ 0.096). These levels of condition are representative of wild cod in the Gulf of St. Lawrence during the spring and during the fall-early winter, respectively. The improved condition did not change mass-specific SMR. However, mass-specific AMR, aerobic scope, and EMR decreased with the improvement in condition. The various measures of swimming performance were affected differently. Ucrit increased and cost of transport at 1.3 and 1.5 body lengths s(-1) decreased with improved condition, but the cost of transport at 0.3, 0.9, 1.1, 1.7, and 1.9 body lengths s(-1), sprint performance, and time to exhaustion did not change. Hierarchies for the speed at first burst-coast, the proportion of Ucrit supported by burst-coasts, and time to exhaustion were maintained with the improvement in condition. The relationships between metabolic rates and swimming performance differed with condition level, with stronger correlations apparent in the cod at their initial condition. Given the low condition of wild cod stocks, these responses indicate that reduced performance, due to both maintenance of metabolic expenditures and modified swimming capacities, may impair survival under conditions of reduced food availability. PMID- 16380933 TI - Trypsin gene expression by quantitative in situ hybridization in carnivorous and herbivorous prickleback fishes (Teleostei: Stichaeidae): ontogenetic, dietary, and phylogenetic effects. AB - We determined trypsin gene sequences and compared relative levels of trypsin gene expression as influenced by ontogeny, diet, and phylogeny in four related prickleback fish species. Of these species, Cebidichthys violaceus and Xiphister mucosus shift from carnivory to herbivory at approximately 45 mm standard length [SL], whereas Xiphister atropurpureus and Anoplarchus purpurescens remain carnivores. Pairwise sequence similarities among the four species were 77%-95% for the trypsin nucleotides and 69%-94% for the amino acids. Trypsin gene expression levels in small (30-40 mm SL) and larger (60-75 mm) wild-caught juveniles and larger (60-75 mm) juveniles raised on a high-protein artificial diet increased with ontogeny in all four species but in response to the diet only in the two carnivores. The indistinguishable expression levels in the sister taxa, X. mucosus and X. atropurpureus, represented the only apparent phylogenetic effect. Xiphister atropurpureus, however, increased both trypsin gene expression and enzymatic activity (the latter from a previous study) on the high-protein artificial diet, indicating transcriptional rather than posttranscriptional (shown in X. mucosus) regulation of the activity. This study provides evidence for genetically programmed upregulation of trypsin gene expression with ontogeny in both the carnivorous and herbivorous species but in response to the high protein artificial diet only in the carnivores. PMID- 16380934 TI - From physiology to fitness: the costs of a defensive adaptation in rattlesnakes. AB - The costs of using and maintaining presumed adaptations are unknown for most animals. Energetically expensive traits, such as some agonistic and antipredator behaviors in animals, may incur trade-offs with other aspects of an animal's life history, such as feeding and reproduction. However, infrequent and brief use may reduce the costs of vigorous behaviors. The shaker muscles in the tails of rattlesnakes are an excellent system for studying the potential costs of a specialized defensive system. The high energetic cost of rattling may increase feeding requirements or use energy that could otherwise be available for reproduction. I used energetic modeling to test whether the cost of rattling in western diamond-backed rattlesnakes (Crotalus atrox) can be high enough to increase feeding demands or reduce fecundity and fitness. Only very frequent and prolonged rattling would increase feeding needs and perhaps reduce fecundity to some degree. Typically, rattling probably incurs very low costs to feeding, reproduction, and hence fitness. These and other results suggest that many seemingly expensive adaptations may have minimal costs to energy budgets, reproduction, and fitness. PMID- 16380935 TI - Both incubation temperature and posthatching temperature affect swimming performance and morphology of wood frog tadpoles (Rana sylvatica). AB - In many oviparous vertebrates, hatchling phenotypes are influenced by egg incubation temperature. Many of those phenotypic traits can also acclimate to long-term thermal conditions of juveniles and adults, yet the interactive effects of prehatching and posthatching temperatures on phenotypes have not been studied. To address such interaction, we incubated eggs of wood frogs (Rana sylvatica) at two temperatures and subsequently reared larvae at three temperatures in a fully factorial design. We measured body size, size-independent morphology, and burst swimming speed at one developmental stage. Body size was independent of egg temperature but decreased significantly with increasing larval temperature. Size independent morphology depended in complex ways on both temperature treatments directly and on their interaction. Burst speed was not influenced directly by egg temperature but was influenced by larval temperature and by the interactions among egg temperature, larval temperature, and test temperature. Our results indicate pervasive effects of egg temperature even late in the larval period and show that prehatching and posthatching temperatures can interact to affect various phenotypic traits. Tadpoles may be able to alter the long-term effects of incubation temperature by choosing particular larval developmental temperatures. Thus, the importance of incubation temperature in oviparous vertebrates should be evaluated by considering the effects of posthatching temperatures. PMID- 16380936 TI - Ontogenetic diet shifts and digestive constraints in the omnivorous freshwater turtle Trachemys scripta. AB - Many reptiles undergo an ontogenetic diet shift from carnivory to herbivory. In this study, we used the yellow-bellied slider turtle, Trachemys scripta, as a model to evaluate whether juvenile turtles are carnivorous because physiological constraints preclude herbivory. We conducted feeding trials in which we fed juvenile and adult turtles a duckweed plant, Lemna valdiviana, or a freshwater grass shrimp, Palaemontes paludosus, for 5 wk. During the trials, we measured mass-specific intake, digestibility, and digestible intake for both size classes, as well as juvenile growth. At the end of the trials, we measured the nutrient composition of the juvenile turtles. Juveniles fed shrimp grew 3.2 times faster than those fed duckweed and had equivalent lipid stores. Digestive processing in juveniles was extremely efficient on the shrimp diet, with higher mass-specific intakes than adults and very high digestibilities (97%). Juveniles digested duckweed as well as adults did; however, their intake of this diet was limited, possibly by the time required for fermentation. We concluded that although juveniles can process plant material, an animal diet allows for greater juvenile growth, which in turtles is linked to higher survivorship and increased future reproductive success. PMID- 16380937 TI - Nectar xylose metabolism in a rodent pollinator (Aethomys namaquensis): defining the role of gastrointestinal microflora using 14C-labeled xylose. AB - The Namaqua rock mouse Aethomys namaquensis, a rodent pollinator of certain geoflorous Protea species, consumes nectar containing xylose. Xylose is not known to be efficiently utilized by mammals. However, it is fermented by certain bacteria, yeasts, and fungi, particularly gastrointestinal bacteria. The end products of microbial fermentation are utilized by the host in oxidative metabolism. Here we investigate the degree to which intestinal bacteria of A. namaquensis contribute to xylose metabolism. Mice were caught during Protea humiflora flowering and nonflowering seasons and given an oral dose of 14C labeled xylose. Exhaled CO2 and excreted urine and feces were continuously collected for 30 h thereafter, and label recovery was determined. Each mouse was then treated with antibiotics to reduce gut microflora, and the experiment was repeated. With their natural gut flora population intact, mice caught during the flowering season exhaled significantly more 14CO2 than did mice caught during the nonflowering season. Also, during both seasons, mice exhaled significantly more 14CO2 before antibiotic treatment than after. Antibiotic treatment caused a significant increase in the proportion of 14C-labeled xylose that was excreted in the urine. The mouse diet likely influences the composition of the gastrointestinal community. Aethomys namaquensis relies on its gut microflora to ferment xylose, thereby converting it into end products that are used by the mice for metabolism. PMID- 16380938 TI - Prioritization or summation of events? Cardiovascular physiology of postprandial Dungeness crabs in low salinity. AB - Decapod crustaceans commonly forage in estuarine environments. The osmoregulatory mechanisms that allow them to cope with periodic episodes of low salinity have been well documented. There is less information on how ventilatory and cardiovascular mechanisms aid survival in low salinity. Prior experiments have shown that most species exhibit a tachycardia coupled with an increase in ventilation rate and oxygen uptake. However, these previous experiments were conducted on animals that were starved before experimentation in order to avoid increases in metabolism associated with digestive processes. This study investigated how the Dungeness crab Cancer magister balances the demands of physiological systems during feeding and digestion in low salinity. Cardiac and ventilatory parameters increased during feeding. When the crabs were subjected to low salinity after feeding, heart rate increased in 25% seawater (SW) but decreased in 50% SW. Instead of an expected increase in ventilation rate during low-salinity exposure, there was a decrease. Feeding was associated with an increase in sternal artery flow, with subsequent decreases in flows through the sternal and anterolateral arteries in low salinity. When low salinity was administered first, a tachycardia occurred, coupled with decreased stroke volume and cardiac output. There was also an increase in ventilation rate. When crabs were fed in low salinity, heart rate decreased in 50% SW but was maintained in 25% SW. Ventilation rate decreased when crabs fed in 50% and 25% SW. Flow through the sternal artery and anterolateral arteries decreased in low salinity, and except for transient increases while feeding, there were further decreases during digestion. Cardiac and ventilatory parameters were rapidly regained when control conditions were restored. The results suggest that events during low salinity are prioritized. Nevertheless, these alterations in physiological parameters may not be beneficial; although digestive processes did not affect osmoregulatory ability, postprandial crabs did not survive as long as starved crabs in 25% SW. The results show that the digestive state of an animal is important in modulating its physiological responses to environmental perturbations, underscoring the importance of an integrative approach to studying physiological responses at the organismal level. PMID- 16380939 TI - Regulation of blood pressure in the land crab Cardisoma guanhumi. AB - We examined the cardiovascular responses to acute and chronic changes in blood volume (BV) in the land crab Cardisoma guanhumi. Acute reduction in BV caused an increase in activity in the dorsoventral muscles (DVMs) and to a lesser extent in the epimeral attractor muscles (EAMs). Contraction of the DVMs and EAMs will decrease the volume of the dorsal sinus and the thorax as a whole, respectively. BV reduction also caused bradycardia with frequent periods of cardiac arrest. There was a small drop in hemolymph pressure. BV expansion had the reciprocal effect on DVM and EAM activity but had no effect on heart rate (fH). After the cardioregulatory nerves were cut, acute hypovolemia had no effect on fH but still caused a moderate increase in DVM activity. After dehydration-induced BV reduction, DVM activity increased, whereas hemolymph pressure, fH, and EAM activity were maintained close to control levels. PMID- 16380940 TI - Changes in temperature and light alter the flight speed of hornets (Vespa crabro L.). AB - Although the principal mechanisms of insect flight are well understood, knowledge about the influence of environmental factors on flight performance is scarce. We show that ambient temperature (TA) and illuminance have an influence on the flight performance of hornets. Moreover, the regulation of flight speed at different environmental conditions is sex specific. In roundabout experiments with tethered hornets, the flight speed of workers was independent of TA, while that of drones was negatively correlated with TA. We hypothesize that the reasons for these differences may lie in sex-specific cooling mechanisms; workers might regulate their body temperature through forced heat loss, whereas drones might reduce their heat production rates. At low illuminance, workers reduced their flight speed, whereas drones did not. Reduced flight speed at low illuminances is probably a behavioral mechanism of compensation for disadvantages caused by changing neuronal processes in the sensory neurons in darkness. PMID- 16380941 TI - A three-dimensional functional assessment of heart and vessel development in the larva of the zebrafish (Danio rerio). AB - There has been considerable recent interest in the development of the circulation in the zebrafish. Optical techniques typically used to visualize changes in heart size allow measurement of stroke volume during early vertebrate development, but this approach is complicated in zebrafish larvae because of the heart's irregular shape and its significant change in morphology during the first 6 d of development. By use of a three-dimensional integration of the early zebrafish heart and vessels, we have greatly reduced measurement error of stroke volume and cardiac output and have determined the cross-sectional growth of major vessels in the developing zebrafish larvae. A dramatic 500%-600% increase in cardiac output (from 10 to 50-60 nL min(-1)) occurs on days 5 and 6 postfertilization in Danio rerio. Cross-sectional area of key vessels (dorsal artery, caudal artery, dorsal vein) as well as between-individual variation significantly decreased over the first 6 d of development. Associated with the decrease in cross-sectional area is a significant increase in red blood cell velocity on days 5 and 6 postfertilization. Together, the three-dimensional data of the cardiac and vascular systems have shown that the most profound physiological and developmental changes occur in days 5 and 6, which corresponds with the appearance of the adult form of the heart and the transition from diffusive to convective O2 supply to internal tissues. PMID- 16380942 TI - Reliability in estimates of body composition of birds: oxygen-18 versus deuterium dilution. AB - Body composition in birds was evaluated indirectly by 18O and 2H dilution. Body composition was determined by whole-body chemical analysis of eight adult roosters (Gallus gallus). In vivo measurements of total body water (TBW) were carried out using doubly labeled water (2H2 18O). Estimated dilution spaces using both the plateau and intercept approaches were compared with the results obtained by carcass lyophilization. Both 18O and 2H slightly overestimated TBW compared with the results obtained by lyophilization, by 2.2%+/-1.9% and 5.7%+/-0.2%, respectively; both differences were statistically significant (P<0.01). The difference between these isotope estimations was significant (P<0.001). However, isotope dilution spaces and TBW were highly correlated. There was a strong inverse relationship between total body fat and TBW percentages (r2=0.98, P<0.0001). The relation between TBW and body protein was significant. Water content in lean body mass (72.8%) obtained in our study was very close to that reported in mammals, demonstrating no fundamental difference in tissue water content between birds and mammals. Estimated body fat and protein values from isotopic dilution did not significantly differ from values obtained by direct chemical analysis (P>0.05), except for body fat in the Pace and Rathbun approach (Table 3). Although estimation of TBW and body composition by isotope dilution is time consuming and expensive, deuterium offers a reliable and low-cost alternative compared with 18O. The advantage of in vivo estimation of TBW with isotopic dilution in combination with the regression approach is that it permits repeated measurements of body composition on the same birds under laboratory and free-living conditions. PMID- 16380943 TI - A search algorithm for fixed-composition protein design. AB - We present a computational protein design algorithm for finding low-energy sequences of fixed amino acid composition. The search algorithms used in protein design typically do not restrict amino acid composition. However, the random energy model of Shakhnovich suggests that the use of fixed-composition sequences may circumvent defects in the modeling of the denatured state. Our algorithm, FC_FASTER, links fixed-composition versions of Monte Carlo and the FASTER algorithm. As proof of principle, FC_FASTER was tested on an experimentally validated, full-sequence design of the beta1 domain of protein G. For the wild type composition, FC_FASTER found a lower energy sequence than the experimentally validated sequence. Also, for a different composition, FC_FASTER found the hypothetical lowest-energy sequence in 14 out of 32 trials. PMID- 16380944 TI - Continuous medium theory for nonequilibrium solvation: IV. Solvent reorganization energy of electron transfer based on conductor-like screening model. AB - In this work the authors present some evidences of defects in the popular continuous medium theories for nonequilibrium solvation. Particular attention has been paid to the incorrect reversible work approach. After convincing reasoning, the nonequilibrium free energy has been formulated to an expression different from the traditional ones. In a series of recent works by the authors, new formulations and some analytical application models for ultrafast processes were developed. Here, the authors extend the new theory to the cases of discrete bound charge distributions and present the correct form of the nonequilibrium solvation energy in such cases. A numerical solution method is applied to the evaluation of solvent reorganization energy of electron transfer. The test calculation for biphenyl-cyclohexane-naphthalene anion system achieves excellent agreement with the experimental fitting. The central importance presented in this work is the very simple and a consistent form of nonequilibrium free energy for both continuous and discrete charge distributions, based on which the new models can be established. PMID- 16380945 TI - FTIR studies on the bond properties of the aspartyl phosphate moiety of the Ca2+ ATPase. AB - As part of our work to determine the bond properties of the aspartyl phosphate moiety of the Ca(2+)-ATPase (SERCA1a) phosphoenzymes, we analyzed Morse potentials of the bridging P-O bond as well as C=O bond strengths for the model compound acetyl phosphate and the two phosphoenzyme intermediates Ca(2)E1P and E2P. Reaction-induced infrared difference spectroscopy was used and a carbonyl band of E2P at 1708 cm(-1) in the presence of mM Mg(2+) was tentatively assigned to the carbonyl group of phosphorylated Asp(351) because of its sensitivity to divalent cations. This band is found at 1716 cm(-1) with mM Ca(2+), for Ca(2)E1P at 1717 cm(-1) with Mg(2+), and at 1719 cm(-1) with Ca(2+) and at 1718 cm(-1) for acetyl phosphate in the absence of divalent cations. The similar band positions indicate similar strengths of interaction of the carbonyl oxygen in acetyl phosphate and the two phosphoenzymes. Together with information on the P-O bond strengths, this implies that the bridging oxygen exerts stronger interactions in the phosphoenzymes than in acetyl phosphate. PMID- 16380946 TI - Discovery of a pseudo beta barrel: synthesis and formation by tiling of ferrocene cyclopeptides. PMID- 16380947 TI - Evidence from NMR and EXAFS studies of a dynamically uniform mononuclear single site zeolite-supported rhodium catalyst. PMID- 16380948 TI - Toward the isolation of functional organic nanotubes. PMID- 16380949 TI - Dimerization of lithiated terminal aziridines. PMID- 16380950 TI - Bond orders between molecular fragments. AB - An extension of the Mayer bond order for the interaction between molecular fragments is presented. This approach allows the classical chemical concepts of bond order and valence to be utilised for fragments and the interactions between the fragments and symmetry-adapted linear combinations to be analysed. For high symmetry systems, the approach allows the contribution from each irreducible representation to be assessed and provides a semiquantitative measure of the role of each bonding mode to interfragment interactions. The utility of this tool has been examined by a study of the bonding in symmetrical sandwich complexes. The validity of the frontier-orbital approach and the contributions from each frontier-orbital interaction can also be assessed within this model. As demonstrated by a study of a number of mixed-sandwich complexes, the model proves to be especially useful for low-symmetry systems in which separation of the sigma, pi and delta roles in bonding of the ligand is difficult to assess. The fragment bond order describes the interaction between preoptimized fragment orbitals and is independent of the charges that are placed on these fragments. Although the method allows the chemist to define fragments in any way they choose, most insight is gained by using the same frontier orbitals employed so successfully in perturbational molecular-orbital approaches. The results are free from the influence of the electron-counting method used to describe fragments, such as the rings and metals in sandwich complexes. PMID- 16380951 TI - Platform synthesis: A useful strategy for rapid and systematic generation of molecular diversity. AB - Emergence of library-based approaches have changed the way of developing new functional molecules in materials science and pharmaceutical science. Therefore, reliable methods for rapid and systematic generation of functional molecules are highly called for in this field. We herein describe our concept of "platform synthesis" as a useful strategy for generating molecular diversity. This simple yet powerful strategy realizes the synthesis of a number of interesting multifunctional molecules, such as multisubstituted olefins, in a programmable and diversity-oriented format. As well as applications to the synthesis of pharmaceutically important molecules, such as tamoxifen and CDP840, applications to materials science, which have led to the discovery of interesting fluorescent materials and properties, are also described. PMID- 16380952 TI - Characterization and catalytic-hydrogenation behavior of SiO2-embedded nanoscopic Pd, Au, and Pd-Au alloy colloids. AB - Colloids embedded in a silica sol-gel matrix were prepared by using fully alloyed Pd-Au colloids, and pure Pd and Au colloids stabilized with tetraalkylammonium bromide following a modified sol-gel procedure with tetrahydrofuran (THF) as the solvent. Tetraethoxysilicate (TEOS) was used as the precursor for the silica support. The molar composition of the sol was TEOS/THF/H2O/HCl = 1:3.5:4:0.05 for the bimetallic Pd-Au and TEOS/THF/H2O/HCl = 1:4.5:4:0.02 for Pd and Au monometallic systems. After refluxing, the colloid was added as a 4.5 wt % solution in THF for Pd-Au, 10.2 wt % solution in THF for Pd and 8.4 wt % solution in THF for Au at room temperature. The gelation was carried out with vigorous stirring (4 days) under an Ar atmosphere. Following these procedures, bimetallic Pd-Au-SiO2 catalysts with 0.6 and 1 wt % metal, and monometallic Pd- and Au-SiO2 catalysts with 1 wt % metal were prepared. These materials were further treated following four different routes: 1) by simple drying, 2) in which the dried catalysts were calcined in air at 723 K and then reduced at the same temperature, 3) in which they were directly reduced in hydrogen at 723 K, and 4) in which the surfactant was extracted using an ethanol-heptane azeotropic mixture. The catalysts were characterized by nitrogen adsorption-desorption isotherms at 77 K, H2 chemisorption measurements, solid-state 1H, 13C, 29Si-CP/MAS-NMR spectroscopy, powder X-ray diffraction (XRD), small angle X-ray scattering (SAXS), X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), and 197Au Mossbauer spectroscopy. The physical characterization by a combination of these techniques has shown that the size and the structural characteristics of the Pd-Au colloid precursor are preserved when embedded in an SiO2 matrix. Catalytic tests were carried out in selective hydrogenation of 3-hexyn-1-ol, cinnamaldehyde, and styrene. These data showed evidence that alloying Pd with Au in bimetallic colloids leads to enhanced activity and most importantly to improved selectivity. Also, the combination of the two metals resulted in catalysts that were very stable against poisoning, as was evidenced for the hydrogenation of styrene in the presence of thiophene. PMID- 16380953 TI - Array formation in evanescent waves. PMID- 16380954 TI - Estimation of Joule heating effect on temperature and pressure distribution in electrokinetic-driven microchannel flows. AB - In this study we present simple analytical models that predict the temperature and pressure variations in electrokinetic-driven microchannel flow under the Joule heating effect. For temperature prediction, a simple model shows that the temperature is related to the Joule heating parameter, autothermal Joule heating parameter, external cooling parameter, Peclet number, and the channel length to channel hydraulic diameter ratio. The simple model overpredicted the thermally developed temperature compared with the full numerical simulation, but in good agreement with the experimental measurements. The factors that affect the external cooling parameters, such as the heat transfer coefficient, channel configuration, and channel material are also examined based on this simple model. Based on the mass conservation, a simple model is developed that predicts the pressure variations, including the temperature effect. An adverse pressure gradient is required to satisfy the mass conservation requirement. The temperature effect on the pressure gradient is via the temperature-dependent fluid viscosity and electroosmotic velocity. PMID- 16380955 TI - Calibration of migration times of variable salinity samples with internal standards in capillary electrophoresis. AB - A practical approach is presented for identifying the analyte peaks stacked by transient ITP (TITP) in samples of uncontrolled salinity. For TITP with chloride ions acting as the leading electrolyte, the effect of matrix chloride of an unknown concentration was calibrated using multiple internal standards to predict the migration times of weakly acidic anionic analytes behaving as strong electrolytes to an accuracy of over 99.9%. The calibration equations for the migration time of an analyte are given as a function of the migration times of internal standards using the mobilities of the relevant ions as parameters. The effects of matrix chloride and various separation conditions such as the temperature, plug length, ionic strength, and pH of the BGE were completely eliminated from the calibration equations. In addition, the actual mobilities, determined for a standard saline sample under the working conditions, were used, and thus, there was no need to conduct supplementary experiments to determine the absolute mobilities at infinite dilution. The internal standards were dyes, which were easily identified in an auxiliary channel monitoring the absorbance at a longer wavelength. For five standard saline matrices containing 100-300 mM NaCl at intervals of 50 mM, the mean absolute error (MAE) in migration times calibrated with two internal standards was 0.4 s (n=5x13). For an electropherogram of a real standard reference urine sample, peaks of spiked analytes were identified with an MAE of 0.9 s (n=13) without conductivity normalizing or desalting of the sample. PMID- 16380956 TI - Loading capacity of carrier ampholytes--based buffers in capillary electrophoresis. AB - Narrow pH cuts of carrier ampholytes (CAs), originally designed for IEF, have been used as BGEs in CE. Their physicochemical properties, rather high buffering capacity and low conductivity, allow very efficient protein separations under high electric field strength. Due to their isoelectric properties, CA BGEs are expected to present a low ionic concentration and consequently a low loading capacity. In this study, we developed a simple method that allows the estimation of the loading capacity of a UV-absorbing BGE by CE. We first characterized in terms of loading capacity, classical ammediol-chromate UV-absorbing BGEs and a 10 mM histidine solution, a classical isoelectric buffer. Then, the loading capacity of four different CA-based BGEs has been assessed. Experimental results have shown that the CA-based buffers were presenting a rather high loading capacity, comparable to classical buffer ones and far higher than the one of the 10 mM histidine solution. PMID- 16380957 TI - The unified equation for the evaluation of first order reactions in dynamic electrophoresis. AB - The unified equation was validated for first order reactions in dynamic CE with a data set of 31 250 elution profiles. Comparison with the results from conventional iterative computer simulation revealed that the unified equation is superior in terms of success rate and precision. The unified equation was applied to determine the cis-trans isomerization rate constants of the angiotensin converting enzyme inhibitor captopril. The separation of the rotational cis-trans isomeric drug has been performed in an aqueous 66 mM citric acid/Tris buffer at pH 3.0 in a 50 cm polyacrylamide-coated fused-silica capillary. Interconversion profiles featuring pronounced plateau formation and peak broadening were observed. Activation parameters DeltaH not equal and DeltaS not equal were obtained from temperature-dependent measurements between 10 and 25 degrees C in 2.5 K steps. From the activation parameters the isomerization barriers of captopril at 37 degrees C under acidic conditions were calculated to be DeltaG not equal trans-->cis=90.6 kJ/mol and DeltaG not equal cis-->trans=84.6 kJ/mol. By comparison of the kinetic data with the results obtained under basic conditions (pH 9.3) a mechanism of isomerization could be proposed. PMID- 16380958 TI - Leukocyte-associated Ig-like receptor-1 has SH2 domain-containing phosphatase independent function and recruits C-terminal Src kinase. AB - Most inhibitory receptors in the immune system contain one or several immunoreceptor tyrosine-based inhibitory motifs (ITIM) and recruit the SH2 domain containing phosphatases SHP-1, SHP-2 and/or SHIP, which are generally believed to be essential for the inhibitory function. However, it has not been systematically investigated whether ITIM-bearing receptors exert their function through alternative interactions. Here we describe that leukocyte-associated Ig-like receptor (LAIR)-1 has inhibitory function in DT40 chicken B cells that lack both SHP-1 and SHP-2. In addition, we found that LAIR-1 did not recruit SHIP upon phosphorylation. Thus, LAIR-1 can function independently from SH2 domain containing phosphatases and must recruit at least one other signaling molecule. Using a yeast-tri-hybrid system, we found that phosphorylated LAIR-1 bound the C terminal Src kinase (Csk). The interaction required the SH2 domain of Csk and phosphorylation of the tyrosine in the N-terminal ITIM of LAIR-1. We propose that Csk is an additional player in the regulation of the immune system by ITIM bearing receptors. PMID- 16380959 TI - Expansion of Valpha24(+)Vbeta11(+) NKT cells from cord blood mononuclear cells using IL-15, IL-7 and Flt3-L depends on monocytes. AB - Human Valpha24(+)Vbeta11(+) NKT cells are a unique T cell population specifically and potently activated by alpha-galactosylceramide (alphaGalCer; KRN7000) presented by CD1d. Here, we present a simple and efficient method for expanding Valpha24(+)Vbeta11(+) NKT cells from human cord blood mononuclear cells (CBMNC) using alphaGalCer in the presence of interleukin (IL)-15, IL-7 and Flt3-L. The addition of alphaGalCer from day 0, compared to its addition from day 8 or day 15, induced a greater expansion of NKT cells. The maximal expansion of NKT cells was observed after 15 days (2300-fold). Thereafter, the number of NKT cells decreased slowly, a decrease that was correlated with the diminution of CD1d positive cells. NKT cell proliferation induced by alphaGalCer was not observed when CD1d-expressing monocytes were depleted from CBMNC, whereas B cell and dendritic cell depletions had no effect. Expanded NKT cells were CD4(+)CD8(-) and secreted both IL-4 and IFN-gamma. In this system, CD3(+) T cells and CD3( )CD56(+) NK cells were also expanded. However, the expansion of NKT cells had no significant functional effect on T and NK cells. This expansion method of CBMNC derived NKT cells is simple and may be helpful for clinical use. PMID- 16380960 TI - Analysis of FOXP3 protein expression in human CD4(+)CD25(+) regulatory T cells at the single-cell level. PMID- 16380964 TI - Analysis of FOXP3 protein expression in human CD4(+)CD25(+) regulatory T cells at the single-cell level. PMID- 16380965 TI - Direct manipulation of activator protein-1 controls thymocyte proliferation in vitro. AB - B cell activating transcription factor (BATF) belongs to the activator protein-1 (AP-1) superfamily of basic leucine zipper transcription factors and forms heterodimers with Jun that possess minimal transcriptional activity. Mice carrying a p56(lck)HA-BATF transgene were created to observe the effects of constitutive expression of this well-characterized AP-1 inhibitor on T cell proliferation. Consistent with the role of AP-1 in promoting the proliferation of many cell types, BATF-transgenic thymocytes proliferate poorly in vitro when stimulated with anti-CD3epsilon and anti-CD28 antibodies or with Concanavalin A. However, when BATF-transgenic thymocytes were stimulated using a standard treatment of PMA and ionomycin, proliferation is normal. The responsiveness to PMA and ionomycin can be attributed to the dramatic disappearance of the hemagglutinin antigen (HA)-tagged BATF protein which is a PKC-dependent process caused by the down-regulation of the p56(lck) proximal promoter coupled with the rapid turnover of the HA-BATF protein. These studies describe conditions of T cell stimulation that negatively influence transcription of the widely used p56(lck) proximal promoter expression cassette. In addition, the unique circumstances of this regulation were exploited to demonstrate that inhibition of AP-1 activity by BATF exerts a direct, and reversible, effect on T cell proliferation in vitro. PMID- 16380966 TI - Dwarfs in ancient Egypt. AB - Ancient Egypt was one of the most advanced and productive civilizations in antiquity, spanning 3000 years before the "Christian" era. Ancient Egyptians built colossal temples and magnificent tombs to honor their gods and religious leaders. Their hieroglyphic language, system of organization, and recording of events give contemporary researchers insights into their daily activities. Based on the record left by their art, the ancient Egyptians documented the presence of dwarfs in almost every facet of life. Due to the hot dry climate and natural and artificial mummification, Egypt is a major source of information on achondroplasia in the old world. The remains of dwarfs are abundant and include complete and partial skeletons. Dwarfs were employed as personal attendants, animal tenders, jewelers, and entertainers. Several high-ranking dwarfs especially from the Old Kingdom (2700-2190 BCE) achieved important status and had lavish burial places close to the pyramids. Their costly tombs in the royal cemeteries and the inscriptions on their statutes indicate their high-ranking position in Egyptian society and their close relation to the king. Some of them were Seneb, Pereniankh, Khnumhotpe, and Djeder. There were at least two dwarf gods, Ptah and Bes. The god Ptah was associated with regeneration and rejuvenation. The god Bes was a protector of sexuality, childbirth, women, and children. He was a favored deity particularly during the Greco-Roman period. His temple was recently excavated in the Baharia oasis in the middle of Egypt. The burial sites and artistic sources provide glimpses of the positions of dwarfs in daily life in ancient Egypt. Dwarfs were accepted in ancient Egypt; their recorded daily activities suggest assimilation into daily life, and their disorder was not shown as a physical handicap. Wisdom writings and moral teachings in ancient Egypt commanded respect for dwarfs and other individuals with disabilities. PMID- 16380967 TI - Sizing the Jurassic theropod dinosaur Allosaurus: assessing growth strategy and evolution of ontogenetic scaling of limbs. AB - Allosaurus is one of the most common Mesozoic theropod dinosaurs. We present a histological analysis to assess its growth strategy and ontogenetic limb bone scaling. Based on an ontogenetic series of humeral, ulnar, femoral, and tibial sections of fibrolamellar bone, we estimate the ages of the largest individuals in the sample to be between 13-19 years. Growth curve reconstruction suggests that maximum growth occurred at 15 years, when body mass increased 148 kg/year. Based on larger bones of Allosaurus, we estimate an upper age limit of between 22 28 years of age, which is similar to preliminary data for other large theropods. Both Model I and Model II regression analyses suggest that relative to the length of the femur, the lengths of the humerus, ulna, and tibia increase in length more slowly than isometry predicts. That pattern of limb scaling in Allosaurus is similar to those in other large theropods such as the tyrannosaurids. Phylogenetic optimization suggests that large theropods independently evolved reduced humeral, ulnar, and tibial lengths by a phyletic reduction in longitudinal growth relative to the femur. PMID- 16380968 TI - Experimental evolution and phenotypic plasticity of hindlimb bones in high activity house mice. AB - Studies of rodents have shown that both forced and voluntary chronic exercise cause increased hindlimb bone diameter, mass, and strength. Among species of mammals, "cursoriality" is generally associated with longer limbs as well as relative lengthening of distal limb segments, resulting in an increased metatarsal/femur (MT/F) ratio. Indeed, we show that phylogenetic analyses of previously published data indicate a positive correlation between body mass corrected home range area and both hindlimb length and MT/F in a sample of 19 species of Carnivora, although only the former is statistically significant in a multiple regression. Therefore, we used an experimental evolution approach to test for possible adaptive changes (in response to selective breeding and/or chronic exercise) in hindlimb bones of four replicate lines of house mice bred for high voluntary wheel running (S lines) for 21 generations and in four nonselected control (C) lines. We examined femur, tibiafibula, and longest metatarsal of males housed either with or without wheel access for 2 months beginning at 25-28 days of age. As expected from previous studies, mice from S lines ran more than C (primarily because the former ran faster) and were smaller in body size (both mass and length). Wheel access reduced body mass (but not length) of both S and C mice. Analysis of covariance (ANCOVA) revealed that body mass was a statistically significant predictor of all bone measures except MT/F ratio; therefore, all results reported are from ANCOVAs. Bone lengths were not significantly affected by either linetype (S vs. C) or wheel access. However, with body mass as a covariate, S mice had significantly thicker femora and tibiafibulae, and wheel access also significantly increased diameters. Mice from S lines also had heavier feet than C, and wheel access increased both foot and tibiafibula mass. Thus, the directions of evolutionary and phenotypic adaptation are generally consistent. Additionally, S-line individuals with the mini-muscle phenotype (homozygous for a Mendelian recessive allele that halves hindlimb muscle mass [Garland et al., 2002, Evolution 56:1,267-1,275]) exhibited significantly longer and thinner femora and tibiafibulae, with no difference in bone masses. Two results were considered surprising. First, no differences were found in the MT/F ratio (the classic indicator of cursoriality). Second, we did not find a significant interaction between linetype and wheel access for any trait, despite the higher running rate of S mice. PMID- 16380969 TI - Ultrastructural investigation of a salivary gland in a centipede: structure and origin of the maxilla I-gland of Scutigera coleoptrata (Chilopoda, Notostigmophora). AB - The maxilla I-gland of Scutigera coleoptrata was investigated using light and electron microscopy methods. This is the first ultrastructural investigation of a salivary gland in Chilopoda. The paired gland opens via the hypopharynx into the foregut and extends up to the third trunk segment. The gland is of irregular shape and consists of numerous acini consisting of several gland units. The secretion is released into an arborescent duct system. Each acinus consists of multiple of glandular units. The units are composed of three cell types: secretory cells, a single intermediary cell, and canal cells. The pear-shaped secretory cell is invaginated distally, forming an extracellular reservoir lined with microvilli, into which the secretion is released. The intermediary cell forms a conducting canal and connects the secretory cell with the canal cell. Proximally, the intermediary cell bears microvilli, whereas the distal part is covered with a distinct cuticle. The cuticle is a continuation of the cuticle of the canal cells. This investigation shows that the structure of the glandular units of the salivary maxilla I-gland is comparable to that of the glandular units of epidermal glands. Thus, it is likely that in Chilopoda salivary glands and epidermal glands share the same ground pattern. It is likely that in compound acinar glands a multiplication of secretory and duct cells has taken place, whereas the number of intermediary cells remains constant. The increase in the number of salivary acini leads to a shifting of the secretory elements away from the epidermis, deep into the head. Comparative investigations of the different head glands provide important characters for the reconstruction of myriapod phylogeny and the relationships of Myriapoda and Hexapoda. PMID- 16380970 TI - Postembryonic development of Tanystylum bealensis (Pycnogonida, Ammotheidae) from Barkley Sound, British Columbia, Canada. AB - All developmental stages of Tanystylum bealensis are described. This is the first complete developmental series of a pycnogonid species collected from a natural, rather than a laboratory-reared population. Development proceeds through a series of nine instars from egg to adult, with major defining characteristics of the instars being the addition of walking legs, loss of chelifores, and modification of larval appendages into adult palps and ovigers. All stages are free-living on the hydroid Plumularia setacea, except for the first instar (protonymphon), which remains on the ovigers of the adult male for a short time after hatching. Development in T. bealensis is compared to development in both the closely related species T. orbiculare and to the more distantly related Achelia alaskensis. PMID- 16380971 TI - Electrostatic interactions of peptides flanking the tyrosine kinase domain in the epidermal growth factor receptor provides a model for intracellular dimerization and autophosphorylation. AB - The mechanism by which ligand-activated EGFR induces autophosphorylation via dimerization is not fully understood. Structural studies have revealed an extracellular loop mediated receptor dimerization. We have previously presented experimental data showing the involvement of a positive 13 amino acid peptide (R645-R657; P13+) from the intracellular juxtamembrane domain (JM) of EGFR important for intracellular dimerization and autophosphorylation. A model was presented that suggest that P13+ interacts with a negative peptide (D979-E991; P13-) positioned distal to the tyrosine kinase domain in the opposite EGFR monomer. The present work shows additional data strengthening this model. In fact, by analyzing protein sequences of 21 annotated ErbB proteins from 9 vertebrate genomes, we reveal the high conservation of peptides P13+ and P13- with regard to their sequence as well as their position relative to the tyrosine kinase (TK) domain. Moreover in silico structure modeling of these ErbB intracellular domains supports a general electrostatic P13+/P13- interaction, implying that the C-terminal of one receptor monomer is facing the TK domain of the other monomer in the receptor dimer and vice versa. This model provides new insights into the molecular mechanism of ErbB receptor activation and suggests a new strategy to pharmacologically interfering with ErbB receptor activity. PMID- 16380972 TI - Progress in the pursuit of therapeutic adenosine receptor antagonists. AB - Ever since the discovery of the hypotensive and bradycardiac effects of adenosine, adenosine receptors continue to represent promising drug targets. First, this is due to the fact that the receptors are expressed in a large variety of tissues. In particular, the actions of adenosine (or methylxanthine antagonists) in the central nervous system, in the circulation, on immune cells, and on other tissues can be beneficial in certain disorders. Second, there exists a large number of ligands, which have been generated by introducing several modifications in the structure of the lead compounds (adenosine and methylxanthine), some of them highly specific. Four adenosine receptor subtypes (A1, A2A, A2B, and A3) have been cloned and pharmacologically characterized, all of which are G protein-coupled receptors. Adenosine receptors can be distinguished according to their preferred mechanism of signal transduction: A1 and A3 receptors interact with pertussis toxin-sensitive G proteins of the Gi and Go family; the canonical signaling mechanism of the A2A and of the A2B receptors is stimulation of adenylyl cyclase via Gs proteins. In addition to the coupling to adenylyl cyclase, all four subtypes may positively couple to phospholipase C via different G protein subunits. The development of new ligands, in particular, potent and selective antagonists, for all subtypes of adenosine receptors has so far been directed by traditional medicinal chemistry. The availability of genetic information promises to facilitate understanding of the drug-receptor interaction leading to the rational design of a potentially therapeutically important class of drugs. Moreover, molecular modeling may further rationalize observed interactions between the receptors and their ligands. In this review, we will summarize the most relevant progress in developing new therapeutic adenosine receptor antagonists. PMID- 16380973 TI - A Process Analytical Technology approach to near-infrared process control of pharmaceutical powder blending. Part III: Quantitative near-infrared calibration for prediction of blend homogeneity and characterization of powder mixing kinetics. AB - The Process Analytical Technology (PAT) initiative, undertaken by the Food and Drug Administration (FDA), paves the way for improvement of drug manufacturing through real-time measurements that allow better process understanding. This study is the third and final Part in a series of studies that represent an integrated approach for real-time blend uniformity assessment using near-infrared (NIR) technology. In this study, the development of a quantitative NIR model for prediction of blending end point is presented. Process signature was built into NIR calibration models by using blend samples that were collected from actual blend experiments under different processing conditions. Evaluation of various calibration algorithms including principal component regression (PCR), partial least squares (PLS), and multi-term linear regression (MLR) was performed. It was found that linear regression, using a single wavelength, yielded optimum calibration and prediction results. The blending profiles predicted by the NIR quantitative model correlated well to those determined by the UV reference analytical method. Characterization of intra-shell versus inter-shell powder mixing kinetics and its implication in sensor positioning was also performed and will be discussed. PMID- 16380974 TI - A Process Analytical Technology approach to near-infrared process control of pharmaceutical powder blending: Part II: Qualitative near-infrared models for prediction of blend homogeneity. AB - The successful implementation of near-infrared spectroscopy (NIRS) in process control of powder blending requires constructing an inclusive spectral database that reflects the anticipated voluntary or involuntary changes in processing conditions, thereby minimizing bias in prediction of blending behavior. In this study, experimental design was utilized as an efficient way of generating blend experiments conducted under varying processing conditions such as humidity, blender speed and component concentration. NIR spectral data, collected from different blending experiments, was used to build qualitative models for prediction of blend homogeneity. Two pattern recognition algorithms: Soft Independent Modeling of Class Analogies (SIMCA) and Principal Component Modified Bootstrap Error-adjusted Single-sample Technique (PC-MBEST) were evaluated for qualitative analysis of NIR blending data. Optimization of NIR models, for the two algorithms, was achieved by proper selection of spectral processing, and training set samples. The models developed were successful in predicting blend homogeneity of independent blend samples under different processing conditions. PMID- 16380975 TI - Physical stability of crystal hydrates and their anhydrates in the presence of excipients. AB - There are few studies in the literature that deal with the effect of excipients on the kinetics of vapor phase induced hydrate-anhydrate phase transformations. The main purpose of this study was to probe the phase stability of hydrate anhydrate systems in the presence of hygroscopic and nonhygroscopic excipients following exposure to either dehydrating or hydrating conditions. Physical mixtures and compacts of model hydrate formers (theophylline and carbamazepine) and excipients (mannitol, microcrystalline cellulose (MCC), polyvinylpyrrolidone (PVP) K12 and K90) were stored at 22 degrees C and varying relative humidities. Raman spectroscopy was used to monitor the kinetics of transformation between hydrate and anhydrate. In general, excipients were found either to have no effect or to promote dehydration. For hydrate formation, excipients could accelerate, retard, or have no influence on hydration kinetics. MCC was found to have only minimal effects on either the dehydration or hydration kinetics of model compounds, whereas mannitol enhanced dehydration but had little effect on hydration. Different PVP grades showed a variety effects: PVPK12 greatly enhanced the dehydration of both theophylline monohydrate (MT) and carbamazepine dihydrate (DC). PVPK90 also enhanced the dehydration of DC, but had a negligible effect on MT. For hydrate formation, PVPK12 was found to have a retarding effect on theophylline anhydrous (AT) transformation, but enhanced the conversion of carbamazepine anhydrous (AC) to DC, PVPK90 also retarded the hydration of AT, but had no effect on AC. Optical microscopy and X-ray powder diffraction studies suggested that PVP (in particular K12), when stored at high RH, was able to result in the partial dissolution of the active pharmaceutical ingredient and hence changed the hydration process from a solid state to a solution-mediated transformation. In summary, the effect of excipients on the kinetics of dehydration and hydration is complex and needs be rationalized in terms of several excipient properties including physical state, chemical composition, and the possibility of specific API-excipient interactions. It is concluded that a multitude of factors will dictate, and often complicate, the final effect of excipients on the phase transformation kinetics of hydrate formers. PMID- 16380976 TI - Vitamin E succinate suppresses prostate tumor growth by inducing apoptosis. AB - Prostate cancer is a major cause of cancer death and morbidity in western countries. However, because of its intrinsic nature of chemoresistance, there is only limited systemic therapy available for the patients. Vitamin E (VE) has been under intensive study as a chemopreventive agent for various types of cancers. Preclinical studies suggest that vitamin E succinate (VES) is the most effective antitumor analogue of VE, yet there are scarce studies of VES in prostate cancer. In this study, we investigated the effects of VES on a panel of prostate cancer cells, and a xenograft model of prostate cancer. Our results indicate that VES significantly inhibited proliferation and induced apoptosis of prostate cancer cell lines in a dose and time dependent manner. The results of microarray analysis followed by real-time RT-PCR and inhibitor analyses indicated that the VES-induced apoptosis is mediated by caspase-4 in prostate tumor cells. In our animal model of prostate cancer in SCID mouse, daily injection of VES significantly suppressed tumor growth as well as lung metastases. These results suggest a potential therapeutic utility of VES for patients with prostate cancer. PMID- 16380977 TI - Ecto-nucleotidase activities in the fat body of Rhodnius prolixus. AB - In this study, we describe the ability of intact fat body of an insect, Rhodnius prolixus, to hydrolyze extracellular ATP. In these fat bodies, the ATP hydrolysis was low in the absence of any divalent metal, and was stimulated by MgCl(2). Both activities (in the absence or presence of MgCl(2)) were linear with time for at least 30 min. In order to confirm the observed nucleotidase activities as ecto nucleotidases, we used an impermeant inhibitor, DIDS (4, 4' diisothiocyanostylbene 2'-2'-disulfonic acid). This reagent inhibited both nucleotidase activities and its inhibitory effect was suppressed by ATP. Both ecto-nucleotidase activities were insensitive to inhibitors of other ATPase and phosphatase activities, such as oligomycin, sodium azide, bafilomycin, ouabain, vanadate, molybdate, sodium fluoride, levamizole, tartrate, p-NPP, sodium phosphate, and suramin. Concanavalin A, activator of some ecto-ATPases, was able to stimulate the Mg(2+)-independent nucleotidase activity, but not the Mg(2+) dependent one. The Mg(2+)-independent nucleotidase activity was enhanced with increases in the pH in the range between 6.4-8.0, but the Mg(2+)-dependent nucleotidase activity was not affected. Besides MgCl(2) , the ecto-ATPase activity was also stimulated by CaCl(2),() MnCl(2), and SrCl(2), but not by ZnCl(2). ATP, ADP, and AMP were the best substrates for the Mg(2+)-dependent ecto nucleotidase activity, and CTP, GTP, and UTP produced very low reaction rates. However, the Mg(2+)-independent nucleotidase activity recognized all these nucleotides producing similar reaction rates, but GTP was a less efficient substrate. The possible role of the two ecto-nucleotidase activities present on the cell surface of fat body of Rhodnius prolixus, which are distinguished by their substrate specificity and their response to Mg(2+), is discussed. PMID- 16380978 TI - Characterization and biochemical analyses of venom from the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae). AB - During parasitism, the ectoparasitic wasp Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) induces a developmental arrest in host pupae that is sustained until the fly is either consumed by developing larvae or the onset of death. Bioassays using fluids collected from the female reproductive system (calyx, alkaline gland, acid gland, and venom reservoir) indicated that the venom gland and venom reservoir are the sources of the arrestant and inducer(s) of death. Infrared spectroscopic analyses revealed that crude venom is acidic and composed of amines, peptides, and proteins, which apparently are not glycosylated. Reversed phase high performance liquid chromatography (HPLC) and sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) confirmed the proteinaceous nature of venom and that it is composed mostly of mid to high molecular weight proteins in the range of 13 to 200.5 kilodaltons (kDa). Ammonium sulfate precipitation and centrifugal size exclusion membranes were used to isolate venom proteins. SDS-PAGE protein profiles of the isolated venom fractions displaying biological activity suggest that multiple proteins contribute to arresting host development and eliciting death. Additionally, HPLC fractionation coupled with use of several internal standards implied that two of the low molecular weight proteins were apamin and histamine. However, in vitro assays using BTI-TN-5B1-4 cells contradict the presence of these agents. PMID- 16380979 TI - Proteome analysis of the silkworm (Bombyx mori. L) colleterial gland during different development stages. AB - The silkworm, Bombyx mori, colleterial gland developed very slowly until 2 days before emergence, then markedly enlarged due to the accumulation of a glue-like substances (mainly including 85% water and 11% proteins). However, the No glue (Ng) mutant female moth secreted only very little glue-like substance and laid loose eggs naturally. High-resolution two-dimensional polyacrylamide gel electrophoresis, followed by computer-assisted analysis, was used to screen the secretory region of colleterial gland protein patterns during different development stages to find quantitative and qualitative difference in protein expression during the pupae and moth stages. More than 700 protein spots were resolved in different developmental stages from the secretory region of the glands and most of the proteins were distributed in the mass range from 30 to 70 kD with pH 4-8. Through comparison and analysis, it was found that 3 proteins were only expressed in the later pupae stage (one or two days before emergence) and moth stage. Furthermore, these proteins were not expressed in the Ng mutant especially actin. There was a great variation of some protein expression volume during the development. Protein spots that changed more than 1.5-fold in expression level (relative to day 9), including 6 spots that were down-regulated and 2 spots that were up-regulated in expression were excised for identification by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Results indicated that actins that participated or regulated the exocytosis of colleterial gland and other differentially expressed proteins might be related to colleterial gland development or the secretion of a glue-like substance. PMID- 16380980 TI - Fruit and vegetable intake and the risk of stomach and oesophagus adenocarcinoma in the European Prospective Investigation into Cancer and Nutrition (EPIC EURGAST). AB - It is considered that fruit and vegetable (F&V) protect against oesophagus and gastric cancer (GC). However, 2 recent meta-analyses suggest that the strength of association on GC seems to be weaker for vegetables than for fruit and weaker in cohort than in case-control studies. No evidence exists from cohort studies about adenocarcinoma of oesophagus (ACO). In 521,457 men and women participating in the EPIC cohort in 10 European countries, information of diet and lifestyle was collected at baseline. After an average of 6.5 years of follow-up, a total of 330 GC and 65 ACO, confirmed and classified by a panel of pathologists, was used for the analysis. We examined the relation between F&V intake and GC and ACO. A calibration study in a sub-sample was used to control diet measurement errors. In a sub-sample of cases and a random sample of controls, antibodies against Helicobacter pylori (Hp) were measured and interactions with F&V were examined in a nested case-control study. We observed no association with total vegetable intake or specific groups of vegetables and GC risk, except for the intestinal type, where a negative association is possible regarding total vegetable (calibrated HR 0.66; 95% CI 0.35-1.22 per 100 g increase) and onion and garlic intake (calibrated HR 0.70; 95% CI 0.38-1.29 per 10 g increase). No evidence of association between fresh fruit intake and GC risk was observed. We found a negative but non significant association between citrus fruit intake and the cardia site (calibrated HR 0.77; 95% CI 0.47-1.22 per 100 g increase) while no association was observed with the non-cardia site. Regarding ACO, we found a non significant negative association for vegetable intake and for citrus intake (calibrated HRs 0.72; 95% CI 0.32-1.64 and 0.77; 95% CI 0.46-1.28 per 100 and 50 g increase, respectively). It seems that Hp infection does not modify the effect of F&V intake. Our study supports a possible protective role of vegetable intake in the intestinal type of GC and the ACO. Citrus fruit consumption may have a role in the protection against cardia GC and ACO. PMID- 16380982 TI - Combined therapy of an established, highly aggressive breast cancer in mice with paclitaxel and a unique DNA-based cell vaccine. AB - Here, we describe the enhanced benefits of treating a highly aggressive breast cancer in mice with a combination of paclitaxel and immunization with a unique DNA-based cell vaccine. An adenocarcinoma was isolated from a spontaneous neoplasm that arose in the mammary gland of a C3H/He mouse (H-2(k)) (SB5b cells). The vaccine was prepared by transfer of genomic DNA-fragments (25 kb) from the breast cancer cells into a mouse fibroblast cell line (LM), modified to enhance its immunogenic properties. As the transferred DNA is integrated, and replicated as the recipient cells divide, the vaccine could be prepared from relatively small numbers of cancer cells (10(7) = 4 mm tumor). SB5b cells were injected into the mammary fat pad of naive C3H/He mice, which are highly susceptible to the growth of the cancer cells. When the tumors reached 3 mm, the mice were injected s.c. with a noncurative dose of paclitaxel. Six days later, when immune competence returned, the mice received the first of 3 weekly s.c. injections of the vaccine. The combined therapy induced robust cellular immunity to the breast cancer, mediated by CD8+ and NK/LAK cells, which resulted in prolonged survival. The immunity was specific, as immunization with a vaccine prepared by transfer of DNA from B16 melanoma cells into the fibroblasts failed to induce immunity to the breast cancer. This type of vaccine raises the possibility that an analogous strategy could be used in the treatment of breast cancer patients at an early stage of the disease. PMID- 16380983 TI - Near infrared thoracoscopy of tumoral protease activity for improved detection of peripheral lung cancer. AB - Improvement in tumor detection using "smart" probes in combination with microcatheter fluorescence thoracoscopy was evaluated in a mouse model. These imaging probes increase in fluorescence intensity after protease activation; cathepsin B is a major activator of the probes used in this study. Lewis lung carcinoma cells were orthotopically implanted in the subpleural lung parenchyma. Two activatable near infrared (NIR) probes with different excitation and emission wavelength were administered intravenously to determine whether wavelength would modulate target to background ratio (TBR). Mice were selectively intubated and thoracoscopy performed. A 0.8 mm outer diameter imaging catheter was used to record simultaneous white-light (anatomic) and NIR (protease expression) images. At both wavelength pairs evaluated (680/700 and 750/780 nm excitation/emission), the intrinsic luminosity differences between tumors and normal lung in uninjected animals was low (p > 0.3 and p = 0.4, respectively and TBR near 1). In mice receiving protease probes IV, tumors were significantly more fluorescent than adjacent lung (p < 0.0005 for 680/700 and p < 0.006 for 750/780) and TBR increased to approximately 9-fold. Confirmatory fluorescence microscopy and immunohistochemistry were similar and revealed that normal lung had very low levels when compared to tumors of cathepsin B and probe fluorescence. In conclusion, protease sensitive imaging probes selective for cathepsin B, imaged with NIR microcatheters, significantly increase the TBR, making small peripheral lung tumors more readily apparent. Such an approach may be a useful adjunct in staging or restaging patients with lung cancer to find minimal disease in the pleural and subpleural space. PMID- 16380984 TI - Cancer incidence in the age range 0-34 years: historical and actual status in Denmark. AB - On the basis of 55 years of continuous cancer registration in Denmark, we present cancer incidence rates, time trends and birth cohort analyses for persons aged 0 34 years. The group of 40,750 cancer patients showed a substantial over representation of males aged 1-24 years. The cancer pattern among young (15-34 years) men was dominated by testicular cancer (35%), lymphomas (14%) and tumors of the brain (13%), while the pattern among young women was governed by invasive cervical cancer (19%), malignant melanoma (15%) and cancer of the breast (12%). In this age range, a positive time trend was seen after 1970, equivalent to average annual percentage increases of 1.9% for men and 1.8% for women, due mainly to markedly increasing trends for testicular cancer, malignant melanoma, brain tumors, thyroid cancer, skin carcinomas and skin sarcoma among men, and for brain tumors, non-Hodgkin lymphoma, malignant melanoma, skin carcinomas and thyroid cancer among women. We saw no clear time trend for breast cancer among women. The cancer pattern among children (0-14 years) was similar to that reported for other white populations. PMID- 16380985 TI - Childhood cancer control: is progress being made? PMID- 16380981 TI - EST-based profiling and comparison of gene expression in the silkworm fat body during metamorphosis. AB - The fat body plays key roles in metabolism and sustenance of growth throughout the life of a silkworm, and thus represents a model tissue for studying development and metamorphosis. Analysis of 18,480 ESTs derived from larval and pupal fat body cDNA libraries allowed characterization of gene expression patterns in the fat body during metamorphosis. By Phrap assembling, 6,814 estimated transcription units (TUs) were generated, 2,673 of which contained 60% of the total ESTs and represented 1,366 distinctive known genes. These genes were classified into 14 categories based on their known or putative functions, and most were found to be involved in metabolism and cell defense. Further comparative analysis of the expression profiles revealed that the gene expression patterns in the larval fat body strikingly differed from those in the pupal fat body. From this, we identified a subset of genes potentially associated with metamorphic events such as the formation or destruction of specific tissues, and simultaneously confirmed the stage-specific expression patterns of several identified genes using RT-PCR examination. This work will provide a valuable resource for studying regulatory mechanisms associated with fat body developmental changes and silkworm metamorphosis. PMID- 16380986 TI - Animal models for the study of squamous cell carcinoma of the upper aerodigestive tract: a historical perspective with review of their utility and limitations. Part A. Chemically-induced de novo cancer, syngeneic animal models of HNSCC, animal models of transplanted xenogeneic human tumors. AB - Understanding the complex histological, genetic and molecular changes that lead to malignant transformation of squamous epithelia of the head and neck will likely guide the development of methods for improved diagnosis, monitoring and treatment of head and neck squamous cell carcinoma (HNSCC). The development and use of animal models that closely mimic the histopathology and molecular pathogenesis of HNSCC in humans would greatly expand the research possibilities and provide a means of testing potential therapeutic agents. However, many available animal models of HNSCC fall short of this objective. In order for investigators to select the appropriate model to answer scientific questions, it is important to understand the benefits and limitations of available animal models for the study of HNSCC. The purpose of this work is to give an overview of the most pertinent animal models of HNSCC, and to discuss future directions of research in this field. PMID- 16380987 TI - Incidence trends and projections for childhood cancer in Ontario. AB - Studies of cancer incidence patterns and trends can provide useful measures of health burden and possible disease etiology, which can aid the planning of cancer care services. This report aims to characterize trends in incidence of childhood cancer, and to assess the implications of these trends by generating incidence projections to 2015. Cancer incidence data were obtained from the database of the Pediatric Oncology Group of Ontario (POGO), which has registered all cancer cases in Ontario since 1985. Annual incidence rates were calculated with census-based population estimates for the 1986-2001 period. Poisson regression models were used to analyze trends, and to calculate projected numbers of cases up to the year 2015. From 1986 to 2001, 5,163 cancer cases occurred among children aged 0 14. Leukemia, CNS tumors and lymphomas were the most common cancers. The number of incident cases increased by 14%, from 296 in 1986 to 336 in 2001. For all cancers, average annual age-standardized rates increased from 147 per million in 1991 to 157 per million in 2001. Over the next 15 years, the 0-14 year population is expected to decrease from 2.28 million in 2000 to 2.13 million in 2015. A marginally statistically significant trend in incidence was projected for all cancers combined (0.5% increase per year p < 0.10) and a statistically significant increase for lymphomas, (1.2% per year 95% CI = 0.0-3.9%). During this period, the number of cases of leukemia and CNS tumors is expected to remain relatively stable. The number of cases of all cancers is expected to increase by 8%, from the average of 320 in 1995 to approximately 347 in 2015. Understanding of these projections will facilitate health care resource planning. PMID- 16380988 TI - Animal models for the study of squamous cell carcinoma of the upper aerodigestive tract: a historical perspective with review of their utility and limitations part B. Transgenic mouse models. PMID- 16380989 TI - Do model results suggest spontaneous regression of breast cancer? PMID- 16380990 TI - Acid decomposition of omeprazole in the absence of thiol: a differential pulse polarographic study at the static mercury drop electrode (SMDE). AB - The reactions of omeprazole, a potent proton pump inhibitor (PPI), were investigated in the absence of a nucleophile. Reactions were monitored, using differential pulse polarography (DPP) at the static mercury drop electrode (SMDE), in solutions buffered to pH values ranging from 2.0 to 8.0. The fast, sensitive, and selective electrochemical technique facilitated to repeat recordings of successive voltammograms [peak current (nA) vs. peak potential (volts vs. Ag/AgCl saturated with 3.0 M KCl)]. The DPP signals of omeprazole and its degradation products, believed to be due to sulfur functional group (the principal site of electrode reaction), gave advantages over the previously employed UV detection technique. The latter primarily relied on pyridine and benzimidazole analytical signals, which are common reaction products of PPI in aqueous acidic solutions. After peak identification, the resulting current (nA) time (s) profiles, demonstrated that omeprazole undergoes degradation to form two main stable compounds, the first is the cyclic sulfenamide (D(+)), previously believed to be the active inhibitor of the H(+), K(+)-ATPase, the second is omeprazole dimer. This degradation is highly dependant on pH. Unlike previous studies which reported that the lifetime of D(+) is few seconds, the cyclic sulfenamide (D(+)) was found to be stable for up to 5-20 min. The results further indicated that omeprazole converts into the cyclic sulfenamide in an irreversible reaction, consequently, D(+) and sulfenic acid (an intermediate which rapidly converts into D(+)) were not interconvertable. The present work suggested that the sulfenic acid is the active inhibitor in vivo. In addition, the omeprazole reactions, in the absence of the thiol, were not as complicated as were previously reported. PMID- 16380991 TI - Fruit consumption reduces the effect of smoking on bladder cancer risk. The Belgian case control study on bladder cancer. AB - We have hypothesized that consumption of fruit and vegetables may be associated with reduced risk of bladder cancer and that this may interact with cigarette smoking and metabolic genetic polymorphisms. A population-based case-control study was performed in the Belgian province of Limburg among 200 cases and 385 controls. Logistic regression was performed to calculate odds ratios (ORs) for bladder cancer occurrence with corresponding 95% confidence intervals (95% CI). Effect modification by smoking was investigated. We also evaluated interaction between fruit intake and GSTM1, GSTT1, NAT2 and SULT1A1 amongst "ever-smokers." Total vegetable intake was not significantly associated with the risk of bladder cancer (OR 1.15, 95% CI: 0.70-1.88 for the highest compared to the lowest tertile). However, total fruit intake was negatively associated with bladder cancer risk (OR 0.61, 95% CI 0.37-0.99 comparing the same tertiles). Among individuals with low daily fruit consumption, ever smokers had a highly increased risk of bladder cancer risk (OR: 4.23, 95% CI: 1.91-9.40). By increasing the daily fruit consumption, the risk of "ever-smokers" for developing bladder cancer decreased, however it remained significant (OR: 2.15; 95CI%: 1.15-4.05). No interaction was identified between the different genotypes and fruit consumption. We conclude that fruit consumption may decrease the effect of smoking on developing bladder cancer. Antioxidants, found in fruit, may protect against the damage caused by free radicals found in cigarette smoke. Metabolic polymorphisms appear not to modify this relation. PMID- 16380992 TI - Cancer risk among Finnish food industry workers. AB - Occupational cancer risks suggested among food industry workers are inconclusive. The objective of our study was to assess associations between different cancers and working in the food industry in Finland. The carcinogenic exposures are mainly inhalatory, and we were therefore interested in respiratory cancers in particular. We followed up a cohort of all economically active Finns born between 1906 and 1945 for 30 million person-years during 1971-95. The 1970 Census data on occupations were linked with data on subsequent incident cancer cases. Standardized incidence ratios (SIR) and 95% confidence intervals (CI) were calculated for each occupation using the economically active population as the reference. A total of 2,526 incident cancer cases were observed. Elevated risks were observed among male food industry workers for pancreatic (SIR=1.50, CI=1.13 1.96) and kidney cancers (1.51, 1.16-1.94). With respect to specific occupations, there was an excess of lung cancer among female bakers (1.38, 1.01-1.85) and laryngeal cancer among male grain millers (2.60, 1.05-5.36). Occupational exposure is unlikely to be a major risk factor for cancer among Finnish workers employed in typical food industry occupations. PMID- 16380993 TI - Selenium binding protein 1 in ovarian cancer. AB - Selenium binding protein 1 (SELENBP1) was identified to be the most significantly down-regulated protein in ovarian cancer cells by a membrane proteome profiling analysis. SELENBP1 expression levels in 4 normal ovaries, 8 benign ovarian tumors, 12 borderline ovarian tumors and 141 invasive ovarian cancers were analyzed with immunohistochemical assay. SELENBP1 expression was reduced in 87% cases of invasive ovarian cancer (122/141) and was significantly reduced in borderline tumors and invasive cancers (p<0.001). Cox multivariate analysis within the 141 invasive cancer tissues showed that SELENBP1 expression score was a potential prognostic indicator for unfavorable prognosis of ovarian cancer (hazard ratio [HR], 2.18; 95% CI=1.22-3.90; p=0.009). Selenium can disrupt the androgen pathway, which has been implicated in modulating SELENBP1 expression. We investigated the effects of selenium and androgen on normal human ovarian surface epithelial (HOSE) cells and cancer cells. Interestingly, SELENBP1 mRNA and protein levels were reduced by androgen and elevated by selenium treatment in the normal HOSE cells, whereas reversed responses were observed in the ovarian cancer cell lines. These results suggest that changes of SELENBP1 expression in malignant ovarian cancer are an indicator of aberration of selenium/androgen pathways and may reveal prognostic information of ovarian cancer. PMID- 16380994 TI - Lifetime occupational and recreational physical activity and risk of benign prostatic hyperplasia. AB - To investigate the relation between occupational and recreational physical activity (PA) in different periods of life and the risk of benign prostatic hyperplasia (BPH), we conducted a hospital-based, case-control study in Italy. The study included 1,369 histologically confirmed BPH and 1,451 controls, admitted to the same hospitals for acute, nonneoplastic diseases. Odds ratios (ORs) and 95% confidence intervals (CIs) of BPH, according to lifetime PA, were obtained by unconditional multiple logistic regression models, including terms for age, study center and education. Compared to the lowest level of occupational PA, the multivariate ORs for BPH for the heavy/strenuous level were 0.6 (95% CI, 0.4-0.8) at age 15-19, 0.6 (95% CI, 0.4-0.8) at age 30-39 and 0.7 (95% CI, 0.5 0.9) at age 50-59. Moreover, compared to <2 hr/week of recreational PA, the ORs for BPH for the highest level (>or=5 hr/week) were 0.5 (95% CI, 0.4-0.7) at age 15-19, 0.6 (95% CI, 0.5-0.8) at age 30-39, and 0.7 (95% CI, 0.5-0.8) at age 50 59. All inverse trends in risk were significant, and no heterogeneity was found by reason of BPH-diagnosis, age at diagnosis, and body mass index (BMI). The inverse association between PA and BPH risk may be due to favorable hormonal correlates of PA, but residual confounding by socioeconomic covariates cannot be excluded. A moderate PA at any ages may help reducing a sizeable number of BPH. PMID- 16380995 TI - Tumor shedding of laminin binding protein modulates angiostatin production in vitro and interferes with plasmin-derived inhibition of angiogenesis in aortic ring cultures. AB - The growth of solid tumors is largely controlled by the process of angiogenesis. A 67 kDa protein, the laminin binding protein (LBP), is shed from malignant cells in significant amounts and binds to laminin-1 (Starkey et al., Cytometry 1999;35:37-47; Karpatova et al., J Cell Biochem 1996;60:226-34). However, the functions of shed LBP are not fully understood. We hypothesize that matrix-bound LBP could modulate local tumor angiogenesis. In support of this hypothesis, we demonstrate that shed LBP exhibits sulfhydryl oxidase-like activities, and modifies the production of angiostatins from plasmin in vitro. The molecular weights of the autocatalytic products of lys-plasmin incubated with LBP in vitro suggest that PMDs (plasmin A chains attached to degraded B chains) (Ohyama et al., Eur J Biochem 2004;271:809-20) are preferentially generated. Using rat aortic ring assays, we also show that shed LBP reverses plasmin-dependent inhibition of vascular outgrowth. To elucidate which LBP region(s) are active in modulating angiogenesis, limited proteolysis experiments were conducted to determine stable rLBP domains likely to fold correctly, and these were cloned, expressed and purified. The stable LBP fragments were tested for binding to laminin-1 and for competition with shed LBP activity in the aortic ring assay. Results of these studies suggest that the active LBP domains lie within the 137 230 amino acid sequence, a region known to contain 2 bioactive sequences. Since this fragment binds to laminin-1 and modulates angiogenesis, it appears likely that binding of shed LBP to matrix laminin-1 is related to its functions in tumor angiogenesis. The findings presented in this manuscript suggest that LBP shedding could provide a useful therapeutic target. PMID- 16380996 TI - Influence of target gene mutations on survival, stage and histology in sporadic microsatellite unstable colon cancers. AB - High-frequency microsatellite unstable (MSI-H) colon tumors develop as a consequence of mutations at repetitive sequences in target genes. TGFBR2 and ACVR2, encoding TGFbeta superfamily receptors, and the proapoptotic gene BAX are frequent targets for frameshift mutation. We analyzed the effect of these mutations on survival and histology in 2 separate cohorts. Forty-eight MSI-H Dukes B2 colon tumors from a cohort of 172 patients had mutations in TGFBR2, BAX and ACVR2 correlated with patient survival. Further, 54 population-based MSI-H colon cancers of all stages from a cohort of 503 patients had mutations correlated with tumor stage, grade and size. Of 44 amplifiable MSI-H Dukes B2 tumors, 70% harbored TGFBR2, 63% BAX and only 4.5% ACVR2 mutations. While mutation alone did not influence survival, concomitant mutation of TGFBR2 and BAX was associated with an improved prognosis in Dukes B2 patients (p=0.05). ACVR2 mutations were more frequent in the second, population-based cohort (stage II: 32.5%, p<0.05). While no target gene mutation correlated with stage in this cohort, poor histological grade and large tumor volume were associated with mutant ACVR2, but not TGFBR2 or BAX mutations, and likely accounts for the lower prevalence of ACVR2 mutations in the first, well-differentiated Dukes B2 cohort. Because target gene mutations did not correlate with stage, they likely occur early in the pathogenesis of MSI-H cancers. Mutations in TGFBR2 and BAX may improve survival in MSI-H Dukes B2 patients, and mutations of ACVR2 may augment histological changes consistent with poor tumor grade that is characteristic of MSI-H colon cancers, and increase tumor size. PMID- 16380997 TI - Mutation analysis of PIK3CA and PIK3CB in esophageal cancer and Barrett's esophagus. AB - Mutation of PIK3CA, the gene coding for the p110alpha catalytic subunit of phosphoinositide 3-kinase (PI3K), has been reported in a limited range of human tumors. We now report that PIK3CA is also mutated in esophageal tumors. Single strand conformational polymorphism (SSCP) and denaturing high-performance liquid chromatography (DHPLC) were used to screen all 20 exons of PIK3CA in 101 samples from 95 individuals with esophageal cancer and/or Barrett's esophagus. Somatic mutation of PIK3CA was detected in 4 of 35 (11.8%) of esophageal squamous cell carcinomas (SCC) and 3 of 50 (6%) adenocarcinomas. No mutations were detected in any of 17 samples of Barrett's esophagus. For PIK3CB, we screened exons 11 and 22, which code for the regions corresponding to the exon 9 and 20 mutational 'hotspots' of PIK3CA. No somatic changes were detected in PIK3CB This study extends previous observations in other tumor types by demonstrating the presence of somatic PIK3CA mutations in both SCC and adenocarcinoma of the esophagus, thus implicating the PI3K pathway in the initiation and/or progression of esophageal cancers. PMID- 16380998 TI - Alcohol consumption and mammographic density in a multiethnic population. AB - This report examined the association between alcohol intake and breast cancer risk as assessed by mammographic densities in a multiethnic population. Information for this analysis was available from 2 previous investigations: a nutritional intervention study (The Breast, Estrogens, and Nutrition Study; BEAN) with 217 premenopausal women and a nested case-control study within The Multiethnic Cohort (MEC) with 1,250 primarily postmenopausal women. On the basis of self-reported alcohol intake from a validated food frequency questionnaire, women were categorized into abstainers (<1 drink/month), low (<1 drink/day) and high (>or=1 drink/day) alcohol consumers. On average, 3 mammograms were available per woman. Using mixed models, we calculated mean percent densities for each alcohol consumption category while adjusting for covariates. Mean alcohol intakes for women in the BEAN study and for cases and controls in the MEC study were 2.0, 2.7 and 1.8 drinks/week, respectively. Overall, the difference in densities between abstainers and the highest alcohol intake category was only 1-2% and the differences were not statistically significant. However, the difference was 3-5% for women aged 55-65 years and for breast cancer cases. In postmenopausal women without hormone replacement therapy (HRT), breast density increased by 2% for each higher alcohol intake category. Breast densities were also elevated for high alcohol consumers on estrogen-only therapy as compared to abstainers and low consumers, whereas combined HRT use was associated with higher mammographic densities independent of alcohol use. In conclusion, despite the low alcohol intake in our population, alcohol consumers had higher percent breast densities than did abstainers. The larger difference in some subgroups suggests that women with certain characteristics may be more susceptible to the influence of alcohol on breast density than others. PMID- 16380999 TI - Extra-tumoral breast tissue in breast cancer patients: variations with steroid contraceptive use. AB - The association between oral contraceptive (OC) use and benign breast changes in extra-tumoral breast tissue was studied histologically in 1,503 breast cancer patients from The WHO Collaborative Study of Neoplasia and Steroid Contraceptives. The occurrence of ductal hyperplasia, ductal atypia, sclerosing adenosis, cysts, apocrine metaplasia, apocrine hyperplasia, apocrine atypia, adenosis, lobular atypia, duct ectasia, calcifications, inflammatory reaction, lactational metaplasia and a high epithelial-stromal ratio was graded semi quantitatively. Prevalence odds ratio (POR) for each histologic variable was calculated by logistic regression analyses. Patients who had ever used OC had lower occurrence of ductal hyperplasia than never users (POR 0.72 (95% CI 0.52 0.99)). Current use and more than 8 years of use was also associated with a lower prevalence of ductal hyperplasia (POR 0.40 (0.20-0.81) and POR 0.33 (0.17-0.64), respectively). Age > 35 years at first use was associated with increased prevalence of ductal carcinoma in situ (POR 2.15 (1.05-4.40)), but not of atypical ductal hyperplasia. Our results show that the effects of OC use on ductal hyperplasia in non-neoplastic breast tissue of breast cancer patients are similar to what others have found in patients with benign breast disease only. The increased prevalence of extra-tumoral ductal carcinoma in situ in breast cancer patients who started OC use at high age may possibly be explained by a longer preinvasive phase in these patients. PMID- 16381000 TI - LIM kinase 1 increases tumor metastasis of human breast cancer cells via regulation of the urokinase-type plasminogen activator system. AB - Mammalian LIM kinase 1 (LIMK1) phosphorylates and inactivates the actin-binding and -depolymerizing factor cofilin and induces actin cytoskeletal changes. LIMK1 is reported to play an important role in cell motility, but the mechanism of induction of cell motility and the role of LIMK1 in tumor growth, angiogenesis and invasion are poorly understood. Here we show that expression of LIMK1 in MDA MB-435 human breast cancer cells enhanced cell proliferation and cell invasiveness and promoted in vitro angiogenesis. Since tumor metastasis requires degradation of the extracellular matrix by the serine protease urokinase type plasminogen activator (uPA), we examined the role of LIMK1 in the regulation of uPA/uPAR system. LIMK1 overexpression in breast cancer cells upregulated the uPA system, increased uPA promoter activity, induced uPA and uPAR mRNA and protein expression and induced uPA secretion. In contrast, cells transfected with the catalytically inactive LIMK mutant D460N-LIMK1 did not exhibit these phenotypic changes. Blocking antibodies against uPA and uPAR suppressed LIMK1-induced cell invasiveness. In addition, LIMK1 overexpression increased tumor growth in female athymic nude mice, promoted tumor angiogenesis and induced metastasis to livers and lungs, possibly by increasing uPA expression in the tumors. Finally, LIMK1 and uPAR were coordinately overexpressed in human breast tumors. These results suggested an important role for LIMK1 signaling in breast cancer tumor growth, angiogenesis and invasion and a regulatory connection between LIMK1 and the uPA system. PMID- 16381001 TI - Ectopic expression of neutrophil gelatinase-associated lipocalin suppresses the invasion and liver metastasis of colon cancer cells. AB - Neutrophil gelatinase-associated lipocalin (NGAL), also known as lipocalin 2, is a 25-kDa lipocalin initially purified from neutrophil granules. It is thought to play a role in regulating cellular growth since its expression is highly upregulated in a variety of proliferative cells such as cancer cells. However, experimental evidence showing a clear causal relationship between NGAL expression and the proliferation of tumor cells is lacking. Here, we found NGAL expression in highly and poorly metastatic colon cancer cell lines of the same genetic origin correlated inversely with the metastatic potential of these cells, which suggests NGAL participates in the metastatic process. To explore the role NGAL plays in tumor growth and metastasis, the KM12SM human colon cancer cell line, which is highly metastatic while showing decreased NGAL expression, was genetically manipulated to overexpress NGAL. The effects of this on tumor growth and liver metastasis were then analyzed using experimental animal models established by injecting BALB/c nude mice with tumor cells subcutaneously or intrasplenically. Ectopic expression of NGAL in the colon cancer cells had little effect on the growth and viability of the tumor cells both in vitro and in vivo. However, NGAL expression not only suppressed the ability of the colon carcinoma cells to invade Matrigel in vitro, it also substantially inhibited liver metastasis in an experimental animal model. Collectively, these results indicate that NGAL may be a candidate metastasis suppressor in colon cancer cells. PMID- 16381002 TI - Hypoxia inducible factor-1alpha is a prognostic marker in premenopausal patients with intermediate to highly differentiated breast cancer but not a predictive marker for tamoxifen response. AB - Hypoxia is common in many solid tumours, including breast cancer. Hypoxia triggers the expression of hypoxia inducible factor-1alpha (HIF-1alpha), and HIF 1alpha has been associated with an impaired prognosis in breast cancer and down regulation of the oestrogen receptor (ER), potentially affecting the treatment efficiency of antioestrogens. The role of HIF-1alpha regarding prognostic and treatment predictive information in breast cancer has not been established and we therefore analyzed HIF-1alpha using immunohistochemistry in a cohort of 377 premenopausal stage II breast cancers arranged in a tissue microarray. The patients were included in a randomized trial with either 2 years of tamoxifen or no adjuvant treatment. The tamoxifen treatment effect could be studied in subgroups of breast cancer and pure prognostic information could be scrutinized for untreated control patients. HIF-1alpha was scored as positive in 24% of the tumours and correlated positively to tumour size, Nottingham histological grade (NHG), Ki-67, Her2 and cyclin E expression and negatively to lymph node status, cyclin D1, ER and PR (progesterone receptor) expression. Surprisingly, there was no difference in tamoxifen response for patients with high or low HIF-1alpha expressing tumours. In lymph node-positive patients as well as NHG 1/2 tumours, high HIF-1alpha protein expression was significantly associated with an impaired recurrence-free survival (p=0.014, 0.018). When analyzing the subgroup of NHG 1/2 tumours, a high HIF-1alpha expression was the only independent significant prognostic marker in multivariate analysis, including standard prognostic markers, suggesting that HIF-1alpha might be a useful prognostic marker in this subgroup of breast cancer, with a rather good but diverse prognosis. PMID- 16381003 TI - Space-time clustering of childhood cancer in great Britain: a national study, 1969-1993. AB - Previous studies have provided evidence that infections may play a part in the aetiology of certain childhood cancers. The finding of space-time clustering indicates the presence of an environmental component to aetiology and is especially supportive of a role for infections. Space-time clustering occurs when excess numbers of cases of a disease are observed within small geographical locations at limited periods of time and this cannot be explained in terms of general excesses in those locations or at those times. To investigate whether infections may be involved in the aetiology of childhood cancer, we have analysed for space-time clustering using a large set of national population-based data from Great Britain for the period 1969-1993. Data were examined by a second-order procedure based on K-functions, with fixed thresholds of closeness in space (0.5 7.5 km) and closeness in time (0.1-1.5 years). Locations were addresses at diagnosis. Tests were repeated, replacing geographical distances with distances to the 19th-33rd nearest neighbours and this provided the primary result for each analysis. There were a total of 32,295 cases of childhood cancer. The analyses showed statistically significant evidence of space-time clustering for acute lymphoblastic leukaemia over the whole age range (p = 0.04), but especially for ages 1-4 years (p = 0.03). There was less statistically significant evidence for total leukaemia (p = 0.048). Significant space-time clustering was also evident for soft tissue sarcomas (p = 0.03) and osteosarcomas (p = 0.02). Results support other evidence suggesting a role for infections in the aetiology of these particular diagnostic groups. PMID- 16381004 TI - Calcitonin stimulates the secretion of urokinase-type plasminogen activator from prostate cancer cells: its possible implications on tumor cell invasion. AB - Calcitonin (CT) is synthesized and secreted in prostate epithelium, and its secretion from malignant prostates is several folds higher than that in benign prostates. CT receptor (CTR) is expressed in malignant prostate epithelium, and its activation increases invasiveness of prostate cancer (PC) cells via activation of protein kinase A. Since the role of urokinase-type plasminogen activator (uPA) in invasion of PC has been established, we tested the hypothesis that CT increases invasion of PC cells by stimulating uPA secretion from PC cells. Exogenously added CT stimulated the secretion of uPA from PC-3M cells in a dose-dependent manner, which was blocked by Rp.cAMP, a competitive inhibitor of protein kinase A. CT stimulated the secretion of MMP-2 and MMP-9 from PC-3M cells, and also increased their invasiveness. Both these actions of CT were blocked by uPA-neutralizing antibodies. Immunofluorescence studies with PC-3M cells suggest that CT stimulated redistribution of cellular uPA to focal adhesion sites, which was further confirmed by co-immunoprecipitation of uPA with focal adhesion kinase (FAK) in response to CT. These results suggest that CT increases invasiveness of PC cells by stimulating PKA-mediated uPA secretion and by redirecting the secreted uPA to focal adhesion sites. The results also suggest that uPA may, at least in part, mediate proinvasive actions of CT on PC cells by stimulating the secretion of gelatinases and degradation of focal adhesion sites. PMID- 16381005 TI - BRAF mutation, CpG island methylator phenotype and microsatellite instability occur more frequently and concordantly in mucinous than non-mucinous colorectal cancer. AB - Mucinous colorectal cancer (CRC) has been reported to have distinct clinicopathological and genetic characteristics. However, the incidence and the relationship among microsatellite instability (MSI), CpG island methylator phenotype (CIMP) and BRAF and KRAS mutations in mucinous and non-mucinous CRC are not known. Activating mutations of BRAF and KRAS and their relationship with MSI and CIMP were examined in 83 sporadic CRC specimens (26 mucinous and 57 non mucinous CRC). MSI, CIMP, BRAF and KRAS mutation were observed in 17, 24, 25 and 36% of the tumors, respectively. BRAF mutation was highly correlated with MSI (p < 0.001) and CIMP (p < 0.001). A higher incidence of MSI (27% vs. 12%), CIMP (38% vs. 18%, p < 0.05) and BRAF mutation (46% vs. 16%, p < 0.01) was observed in mucinous CRC. KRAS mutation (27% vs. 40%) was observed more frequently in non mucinous CRC. Significantly higher percentages of mucinous CRC (54%, p < 0.05) had MSI or CIMP or BRAF mutations. Concordant occurrence of 2 or more of these alterations was observed in 39% of mucinous CRC and only 11% of non-mucinous CRC (p < 0.01). The more frequent occurrence and closer association among MSI, CIMP and BRAF mutation in mucinous CRC observed in our study further supports the idea that its pathogenesis may involve distinct genetic and epigenetic changes. PMID- 16381006 TI - Tumour-cell apoptosis after cisplatin treatment is not telomere dependent. AB - Cisplatin is a major chemotherapeutic agent, especially for the treatment of neuroblastoma. Telomeres with their sequence (TTAGGG)n are probable targets for cisplatin intrastrand cross-linking, but the role of telomeres in mediating cisplatin cytotoxicity is not clear. After exposure to cisplatin as single dose or continuous treatment, we found no loss of telomeres in either SHSY5Y neuroblastoma cells (telomere length, approximately 4 kbp), HeLa 229 cells (telomere length, 20 kbp) or in the acute lymphoblastic T cell line 1301 (telomere length, approximately 80 kbp). There was no induction of telomeric single strand breaks, telomeric overhangs were not degraded and telomerase activity was down-regulated only after massive onset of apoptosis. In contrast, cisplatin induced a delayed formation of DNA strand breaks and induced DNA damage foci containing gamma-H2A.X at nontelomeric sites. Interstitial DNA damage appears to be more important than telomere loss or telomeric damage as inducer of the signal pathway towards apoptosis and/or growth arrest in cisplatin-treated tumour cells. PMID- 16381007 TI - Contact of high-invasive, but not low-invasive, melanoma cells to native collagen I induces the release of mature cathepsin B. AB - Metastasis of malignant tumor cells involves cell-cell and cell-matrix interactions, which regulate the expression and localization of proteolytic enzymes. In the present study, we investigated the expression and localization of the lysosomal cysteine proteinase cathepsin B and its natural inhibitors cystatin A, B and C in high- (MV3), intermediate- (SKmel28) and low-invasive (SKmel23, WM164) human melanoma cell lines grown on plastic or in contact with monomeric or fibrillar collagen type I. Neither the transcript levels of cathepsin B nor those of the natural inhibitors, cystatin B and C, were altered by the interaction of melanoma cells with collagen type I. However, protein expression and cellular localization of cathepsin B and its inhibitors were markedly affected. In contrast to low-invasive cells, high-invasive cells constitutively released procathepsin B when cultured on plastic. In addition, contact of invasive cells with fibrillar collagen type I resulted in the release of both mature forms of the protease. Perturbation studies using inhibitory antibodies against the beta1 subunit of the integrin receptor indicated a role for the beta1 integrin receptor family in the regulation of cathepsin B release. Cystatin B protein expression was much lower in high-invasive cells in both culture conditions, when compared to low-invasive cells. Cystatin C expression was comparable in all cells, but cell contact to fibrillar collagen type I induced its expression. These results strongly implicate a pivotal role of cell-matrix interactions for the regulation of cathepsin B localization and activity in melanoma cells. PMID- 16381008 TI - Are mammotropic hormones mainly permissive for the development of breast cancer? AB - In a case-control study nested within the Greek EPIC cohort, serum levels of estrone, estradiol, androstenedione, dehydroepiandosterone sulfate, testosterone and IGF-1 were measured for 29 breast cancer patients and 58 control women, matched for age and menopausal status. There was little difference in breast cancer risk when values of 4-6-as compared to values of 1-3-hormones were elevated, a finding arguing against a positive interaction among these hormones. Breast cancer risk, however, was significantly and substantially lower among women with levels of all hormones below the corresponding age- and menopausal status-predicted means, compared to women with levels of at least 1 hormone above the predicted mean (odds ratio = 0.11 with 95% confidence interval 0.01-0.90; p = 0.04). Our results suggest that the studied mammotropic hormones may act as permissive factors for breast cancer occurrence, and that the levels of some of them above the mean suffice for sustaining growth of a developing tumor. A corollary is that studies of mammotropic hormones in relation to breast cancer risk may also need to focus on the lower end of the distributions of these growth enhancing hormones. PMID- 16381009 TI - Inhibition of human prostate cancer growth, osteolysis and angiogenesis in a bone metastasis model by a novel mechanism-based selective gelatinase inhibitor. AB - Metastasis to the bone is a major clinical complication in patients with prostate cancer (PC). However, therapeutic options for treatment of PC bone metastasis are limited. Gelatinases are members of the matrix metalloproteinase (MMP) family and have been shown to play a key role in PC metastasis. Herein, we investigated the effect of SB-3CT, a covalent mechanism-based MMP inhibitor with high selectivity for gelatinases, in an experimental model of PC bone metastases. Intraperitoneal (i.p.) treatment with SB-3CT (50 mg/kg) inhibited intraosseous growth of human PC3 cells within the marrow of human fetal femur fragments previously implanted in SCID mice, as demonstrated by histomorphometry and Ki-67 immunohistochemistry. The anti-osteolytic effect of SB-3CT was confirmed by radiographic images. Treatment with SB-3CT also reduced intratumoral vascular density and bone degradation in the PC3 bone tumors. A direct inhibition of bone marrow endothelial cell invasion and tubule formation in Matrigel by SB-3CT in vitro was also demonstrated. The use of the highly selective gelatinase inhibitors holds the promise of effective intervention of metastases of PC to the bone. PMID- 16381010 TI - Decreased expression of phosphorylated JNK in breast infiltrating ductal carcinoma is associated with a better overall survival. AB - Phosphorylation/activation of c-jun NH2-terminal kinase (JNK) has an ambivalent role, pro-proliferation or antiproliferation, in human cancers, which is determined by different cell types and by its crosstalk with other kinases. So far, the role of phosphorylated JNK (p-JNK) in breast cancer is mostly undefined. In this study, we analyzed the expression of p-JNK, as well as p-ERK1/2 and p-38, in the pair of cancer and noncancer breast tissues, by using immunoblotting techniques. These results were further correlated with the clinicopathological characteristics and overall survival. Decreased p-JNK1/2 expression in cancer tissues was observed in 48.5% of breast infiltrating ductal carcinoma (IDC) cases and was correlated significantly with the increased tumor grade and the decreased age at diagnosis (p = 0.030 and 0.029). Interestingly, the Kaplan-Meier survival curve showed that the decreased p-JNK1/2 expression was associated with a better overall survival of IDC (p = 0.004). The expression of p-JNK1/2 was positively correlated with p-p38 (p = 0.002), but not p-ERK1/2. Furthermore, co-expressed p JNK1/2 and p-p38 was associated with a poor overall survival of IDC (p = 0.007). In conclusion, our results indicate that the aberrant p-JNK1/2 expression and the co-expressed p-JNK1/2 and p-p38 in breast tissues may play a role in the carcinogenesis of breast IDC. PMID- 16381011 TI - The Salzburg concept of intraoperative radiotherapy for breast cancer: results and considerations. AB - Aim of this study is to show that ipsilateral breast tumor recurrence (IBTR) after breast conserving surgery can be reduced by proper surgery and modern radiotherapy techniques. Three hundred and seventy eight women with stage I or II breast cancer had breast conserving surgery and received 51-56.1 Gy of postoperative radiation to the whole breast in 1.7 Gy fractions, but patients received different boost strategies. Group 1 (n = 188) received electron boost radiation of 12 Gy subsequent to the irradiation to the whole breast, group 2 (n = 190) received intraoperative electron boost radiation of 9 Gy directly to the tumor bed, followed by whole breast irradiation. After a median follow up period of 81.0 months in group 1 and a median follow up period of 51.1 months in group 2, 12 IBTRs (6.4%) could be observed in group 1 and no IBTR could be observed in group 2 (0.0%). The 5-year actuarial rates of IBTR were 4.3% (95% CI, 1.9-8.3%) and 0.0% (95% CI, 0.0-1.9%), respectively (p = 0.0018). The 5-year actuarial rates of distant recurrence were 8.6% (95% CI, 4.9-13.5%) and 4.2% (95% CI, 1.8 8.2%), respectively (p = 0.08). The 5 year disease-free survival rates were 90.9% (95% CI, 85.8-94.7%) in group 1 and 95.8% (95% CI, 91.8-98.2%) in group 2 (p = 0.064). Immediate IORT-boost and whole breast irradiation yields excellent local control at 5 years, and was associated with a statistically significant decreased rate of IBTR compared with a similar cohort of patients treated with whole breast irradiation and conventional electron boost. PMID- 16381012 TI - Msh2 deficiency increases susceptibility to benzo[a]pyrene-induced lymphomagenesis. AB - DNA mismatch repair (MMR) is essential for repair of single-base mismatches and insertion/deletion loops. MMR proteins also participate in cellular response to DNA damaging agents such as various alkylating agents. Mice deficient in the MMR gene Msh2 develop tumors earlier after exposure to alkylating agents when compared to unexposed mice. The interaction between the MMR system and polycyclic aromatic hydrocarbons such as benzo[a]pyrene (B[a]P) has not been investigated in vivo. Here, we show that treatment of Msh2-deficient mice with B[a]P enhances susceptibility to lymphomagenesis. Carrying at least one intact copy of the Msh2 gene had a protective effect. B[a]P treatment only induced lymphomas in 3 of the 40 (7.5%) mice with at least one intact copy of the Msh2 gene as compared to 13 of the 17 (76.5%) Msh2-deficient mice and occurs only after a much longer time period. The B[a]P-DNA adduct levels measured in lung, liver, spleen and forestomach of B[a]P-treated Msh2-/- mice were not significantly different from B[a]P-treated Msh2+/+ mice. In summary, the results suggest that B[a]P accelerates lymphomagenesis in Msh2-deficient mice. Furthermore, Msh2 deficiency does not have any significant effect on B[a]P-DNA adduct levels. PMID- 16381013 TI - Gastrointestinal transit and mucoadhesive characteristics of complexation hydrogels in rats. AB - The aim of this study was to investigate the gastrointestinal (GI) transit and mucoadhesive properties of complexation hydrogels, poly(methacrylic acid-grafted ethylene glycol). The fluorescent labeled complexation hydrogels containing different molar ratios of methacrylic acid/ethylene glycol and different particle diameters were synthesized by free radical solution polymerization. The GI transit profiles of microparticles after oral administration to rats were evaluated by determining the polymer remaining in the stomach and the small intestine. Moreover, the mucoadhesion to the duodenal mucosa was evaluated by an in situ perfusion method. The ethylene glycol content and particle size of the hydrogels influenced significantly the GI transit and mucoadhesive capacities. The microparticles composed of polymers prepared from 1:1 ratio of methacrylic acid/ethylene glycol and having diameters of <53 microm showed the strongest mucoadhesive capacity. These findings indicated that the hydrogels may be a promising tool for improving oral bioavailability of various drugs, which are poorly absorbed from the GI tract. PMID- 16381014 TI - A Process Analytical Technology approach to near-infrared process control of pharmaceutical powder blending. Part I: D-optimal design for characterization of powder mixing and preliminary spectral data evaluation. AB - Experimental design, multivariate data acquisition, and analysis in addition to real time monitoring and control through process analyzers, represent an integrated approach for implementation of Process Analytical Technology (PAT) in the pharmaceutical industry. This study, which is the first in a series of three parts, uses an experimental design approach to identify critical factors affecting powder blending. Powder mixtures composed of salicylic acid and lactose were mixed in an 8 qt. V-blender. D-optimal design was employed to characterize the blending process, by studying the effect of humidity, component concentration, and blender speed on mixing end point. Additionally, changes in particle size and density of powder mixtures were examined. A near-infrared (NIR) fiber-optic probe was used to monitor mixing, through multiple optical ports on the blender. Humidity, component concentration, and blender speed were shown to have a significant impact on the blending process. Furthermore, humidity and concentration had a significant effect on particle size and density of powder mixtures. NIRS was sensitive to changes in physicochemical properties of the mixtures, resulting from process variables. Proper selection of NIR spectral preprocessing is of ultimate importance for successful implementation of this technology in the monitoring and control of powder blending and is discussed. PMID- 16381015 TI - Factors influencing regional differences in intestinal absorption of UK-343,664 in rat: possible role in dose-dependent pharmacokinetics. AB - The objective of this study was to evaluate potential contributions of intestinal export and metabolism to the oral dose-dependent pharmacokinetics of the human cGMP-specific phosphodiesterase type 5 inhibitor, UK-343,664. Differences between jejunal and ileal handling of this CYP3A and P-gp substrate were investigated. CYP3A and P-gp display differing activities in the upper and lower mammalian small intestine and their impact on variable drug absorption can be mechanistically assessed for individual compounds with in situ perfusion of rat's small intestine. Isolated segments of rat jejunum and ileum were perfused with UK 343,664 solution and measurements were made as a function of drug concentration for dose dependence and in the presence of CYP3A and P-gp inhibitors. Intestinal permeability and metabolism were measured by total drug disappearance and major metabolite, UK-347,334 (N-desethyl metabolite), appearance in the intestinal lumen. Intestinal tissue and mesenteric blood measurements of drug and metabolite were also determined. The effective permeability (P(eff)) of UK-343,664 and metabolite formation (F(met)) increased as a function of concentration. Regional differences in P(eff) and F(met) were observed with low-intestinal metabolism of UK-343,664 in both regions (<10%). P-gp inhibition caused significant increase in P(eff) and F(met) in jejunum and ileum while ketoconazole, a P-gp and CYP3A inhibitor, has only limited effect on metabolism. In conclusion, UK-343,664 absorption is mainly regulated by P-gp in jejunum and ileum while CYP3A intestinal metabolism has minimal effect. This role of P-gp could explain the dose-dependent pharmacokinetics of UK-343,664 and its unusual behavior of t(max) as a function of dose. PMID- 16381016 TI - Monitoring of disseminated tumor cells in bone marrow in high-risk breast cancer patients treated with high-dose chemotherapy. AB - The present study aimed to investigate the clinical relevance of disseminated tumor cells (DTC) in breast cancer patients before and after high-dose adjuvant chemotherapy with or without progenitor stem-cell support. One hundred and eighteen high-risk stage II breast cancer patients entering the Scandinavian Study Group multicenter trial were randomized to 9 cycles of tailored and dose escalated FEC (5-fluorouracil, epirubicin, cyclophosphamide) or 3 cycles of standard FEC followed by high-dose chemotherapy. Bone marrow (BM) samples at diagnosis and 6 months after completion of chemotherapy were assessed for the presence of cytokeratin positive (CK+) cells. Before treatment, 29% of the patients were CK+ (21% in the dose-escalated group and 36% in the high-dose group). Six months after treatment, 17% of the patients were CK+ (17 and 16% respectively). Of the 95 patients who were evaluated 6 months after treatment, 60% were consistently CK-. CK+ cells in BM was evaluated as a prognostic and predictive marker and compared to other defined prognostic factors of the primary tumor. Monitoring BM changes at the time of diagnosis and 6 months posttreatment is an independent predictive factor for breast-cancer-specific survival (BCS) (p = 0.001). Those who have consistent CK negative (-) BM findings constitute a group of patients with good prognosis. Our results suggest that changes in CK+ cells in BM before and after chemotherapy can be used clinically as a surrogate maker to predict outcome in breast cancer patients. PMID- 16381017 TI - Polymorphisms in the growth hormone receptor: a case-control study in breast cancer. AB - The human growth hormone receptor (GHR) mediates the effects of growth hormone (GH1), starting a signalling cascade that is involved in the regulation of proliferation, differentiation and apoptosis. Recently, an isoform of the GHR gene lacking exon 3 (GHRd3) was associated with accelerated responsiveness to growth hormone. In this study, we investigated the association of the GHRd3 polymorphism with breast cancer risk and performed a haplotype analysis with 3 additional single nucleotide polymorphisms (SNPs) (Gly186Gly, Cys440Phe and Ile544Leu) in the GHR coding region in a Polish cohort. We did not observe any effect of the 4 polymorphisms on breast cancer risk. Neither did the 3 most common haplotypes influence breast cancer risk. However, a rare haplotype (dGGC), containing the GHRd3 allele, was associated with a decreased breast cancer risk (OR 0.30, 95% CI 0.11-0.80). PMID- 16381018 TI - Gene expression profiling identifies potential relevant genes in alveolar rhabdomyosarcoma pathogenesis and discriminates PAX3-FKHR positive and negative tumors. AB - We analyzed the expression signatures of 14 tumor biopsies from children affected by alveolar rhabdomyosarcoma (ARMS) to identify genes correlating to biological features of this tumor. Seven of these patients were positive for the PAX3-FKHR fusion gene and 7 were negative. We used a cDNA platform containing a large majority of probes derived from muscle tissues. The comparison of transcription profiles of tumor samples with fetal skeletal muscle identified 171 differentially expressed genes common to all ARMS patients. The functional classification analysis of altered genes led to the identification of a group of transcripts (LGALS1, BIN1) that may be relevant for the tumorigenic processes. The muscle-specific microarray platform was able to distinguish PAX3-FKHR positive and negative ARMS through the expression pattern of a limited number of genes (RAC1, CFL1, CCND1, IGFBP2) that might be biologically relevant for the different clinical behavior and aggressiveness of the 2 ARMS subtypes. Expression levels for selected candidate genes were validated by quantitative real-time reverse-transcription PCR. PMID- 16381019 TI - The norepinephrine-driven metastasis development of PC-3 human prostate cancer cells in BALB/c nude mice is inhibited by beta-blockers. AB - The development of metastases is a decisive step in the course of a cancer disease. The detection of metastases in cancer patients is correlated with a poor prognosis, and over 90% of all deaths from cancer are not due to the primary tumor, which often can be successfully treated, but are due to the metastases. Tumor cell migration, a prerequisite for metastasis development, is not merely genetically determined, but is distinctly regulated by signal substances of the environment including chemokines and neurotransmitters. We have shown previously that the migration of breast, prostate, and colon carcinoma cells is enhanced by the stress-related neurotransmitter norepinephrine in vitro, and that this effect can be inhibited by the beta-blocker propranolol. We now provide for the first time evidence for the in vivo relevance of this neurotransmitter-driven regulation using PC-3 prostate carcinoma cells. The development of lumbar lymph node metastases in athymic BALB/c nude mice increased with the application of norepinephrine via microosmotic pumps, while propranolol inhibited this effect. However, the growth of the primary tumor was not affected by either treatment. Additionally, experiments using human tissue microarrays showed that 70-90 percent of breast, colon, and prostate carcinoma tissues express the relevant beta2-adrenoceptor. Thus, our work contributes to the understanding of the basic cellular mechanisms of metastasis development, and furthermore delivers a rationale for the chemopreventive use of clinically established beta-blockers for the inhibition of metastases. PMID- 16381020 TI - Cancer incidence in a cohort of Ontario and Quebec women having bilateral breast augmentation. AB - The possibility that women, who receive breast implants for cosmetic purposes, have increased long-term risks of developing cancer continues to be debated. The objective of our study was to prospectively examine cancer incidence among women who received breast implants. A cohort was assembled of 24,558 women, 18 years of age and older, who underwent bilateral cosmetic breast augmentation, and 15,893 women who underwent other cosmetic procedures in Ontario or Quebec between 1974 and 1989. These plastic surgery patients were selected from the same clinics as the implant population. Incident cancers were identified by linking to Canadian registry data up to December 31, 1997. In total, 676 cancers were identified among women who received breast implants compared to 899 expected based on general population rates (standardized incidence ratio (SIR) = 0.75; 95% confidence interval (CI) = 0.70-0.81). Overall cancer incidence rates among women who received breast implants were similar to that of the other plastic surgery patients (relative risk (RR) = 0.91, 95% CI = 0.81-1.02). However, women who received breast implants had lower breast cancer rates than the plastic surgery patients (RR = 0.64, 95% CI = 0.53-0.79). No increased risks were observed among the implant population for any of the other cancer sites examined. Comparisons involving only women who received breast implants found no association between long-term breast cancer incidence and implant site (submuscular vs. subglandular), fill (saline vs. silicone) or envelope (polyurethane-coated or not). In conclusion, women undergoing cosmetic breast augmentation do not appear to be at an increased long-term risk of developing cancer. PMID- 16381021 TI - Urgency degree and bladder contraction velocity: sequential changes in women with idiopathic detrusor overactivity. AB - AIMS: In patients with idiopathic detrusor overactivity (DO) who showed storage symptoms worsening with time, we checked whether and which urodynamic parameter changes are associated with an increased urgency degree. MATERIALS AND METHODS: We analyzed retrospectively the urodynamic findings in 54 women -27 with storage symptoms (Group A, mean age 32 +/- 7 years) and 27 controls (Group B, mean age 30 +/- 9 years). These latter had a history of recurrent UTIs (urinary tract infections), but when seen by us had no UTI, DO, lower urinary tract symptoms, or any other pathological finding. Group A had a first urodynamic examination when first referred (time 1) and were re-assessed a mean of 16 months later (time 2) for worsened storage symptoms. RESULTS: In Group A, an idiopathic DO was shown at both times 1 and 2; urgency of voiding could be delayed during cystometry for >or=2 min (= moderate urgency) at time 1 and for <2 min (= severe urgency) at time 2; detrusor contraction strength or contractility proved higher than in Group B (P < 0.001) and increased from time 1 to time 2 (P < 0.001), detrusor shortening velocity being always the major component of the higher contractility levels. CONCLUSIONS: A DO-related increase in bladder contractility may have been further enhanced by severe urgency through a positive feedback mechanism. The urgency degree proved closely associated in DO patients with the level of detrusor shortening velocity rather than with detrusor pressure. PMID- 16381022 TI - CYP17 MspA1 polymorphism and risk of biliary tract cancers and gallstones: a population-based study in Shanghai, China. AB - Biliary tract cancers, encompassing cancers of the gallbladder, extrahepatic bile duct and ampulla of Vater, are rare but highly fatal malignancies. Other than gallstones, little is known about the risk factors for biliary tract cancers. Endogenous estrogens are thought to play a role in the etiology of gallstones and gallbladder cancer, since both conditions predominate in females and are associated with parity and obesity. In view of reports linking the CYP17 MspA1 polymorphism to high circulating levels of estrogens and a predisposition to other hormonally related cancers, we examined the relationship between CYP17 MspA1 variants and risk of biliary disease in a population-based case-control study in Shanghai. The study included 446 cancer cases (254 gallbladder, 139 extrahepatic bile duct, 53 ampullary cancers), 929 biliary stone cases (691 gallbladder, 238 bile duct) and 818 population controls. Genomic DNA from peripheral blood lymphocytes was used for genotyping. Relative to those with the A2/A2 genotype, A1 carriers (A1/A1 and A1/A2 genotypes) had an increased risk of gallbladder cancer (odds ratio (OR) = 1.5, 95% confidence interval (CI) = 1.1 2.1). In addition, women with the A1 allele and high parity (> or =3) had a 3 fold risk of gallbladder cancer (OR = 3.3, 95% CI = 1.6-6.9), compared to those with the A2/A2 genotype and lower parity, with the highest risk seen for those also having biliary stones (OR = 4.6, 95% CI = 1.8-11.7, P(interaction) = 0.04). The A1 allele was not associated with a higher risk of gallstones except among those with body mass index (BMI) greater than 25 kg/m2 (OR = 3.1, 95% CI = 2.0 4.8, P(interaction) = 0.02) and among those with a history of diabetes (OR = 2.5, 95% CI = 1.4-4.3, P interaction = 0.09). No clear relation was seen between the CYP17 polymorphism and cancers of the bile duct or ampulla of Vater. The association of the CYP17 MspA1 polymorphism with an increased risk of gallbladder cancer, as well as biliary stones among overweight and diabetic individuals, suggests an interplay between genetic and hormonal risk factors in gallbladder disease. PMID- 16381023 TI - Variable number of tandem repeats polymorphism in the SMYD3 promoter region and the risk of familial breast cancer. PMID- 16381024 TI - Effects of novelty on behavior in the adolescent and adult rat. AB - Adolescence is a time of high-risk behavior and increased exploration. This developmental period is marked by a greater probability of initiating drug use and is associated with an increased risk to develop addiction and dependency in adulthood. Human adolescents are predisposed towards an increased likelihood of risk taking behaviors (Zuckerman, 1986), including drug use or initiation. The purpose of the study was to examine differences in developmental risk taking behaviors. Adolescent and adult animals were exposed to a novel stimulus in a familiar environment to assess impulsive behaviors, novelty preference, and exploratory behaviors. Adolescent animals had greater novelty-induced locomotor activity, greater novelty preference, and showed higher approach and exploratory behaviors compared to adult animals. These data support the notion that adolescents may be predisposed toward sensation seeking and consequently, are more likely to engage in risk-taking behaviors, such as drug use initiation. PMID- 16381025 TI - Fibronectin as target for tumor therapy. AB - During cancer progression, the extracellular matrix (ECM) of the tissue in which the tumor grows is extensively remodeled, both by degradation of preexisting ECM molecules and by the neosynthesis of ECM components, which in many cases are not present in the ECM of normal tissues. Fibronectin (FN), a class of high-molecular weight adhesive glycoproteins, plays a prominent role in mediating ECM function, because of its high abundance and its interaction with cellular components. Furthermore, the generation of tumor-associated FN isoforms allows the development of specific ligands (e.g., antibodies), which can be used for the selective delivery of therapeutic agents to the tumor environment. In view of these considerations, it is not surprising that FN is being used as a target for biomolecular intervention, both for the development of inhibitory molecules that block the interaction of FN with integrins and other receptors on the cell surface, and for the development of ligand-based targeted imaging and therapeutic strategies. In this review, we briefly present the essential properties of FN, and we then focus on the therapeutic strategies that are currently in preclinical or clinical development and feature FN as a target, or that are based on FN fragments so as to promote tumor-growth inhibition. PMID- 16381026 TI - Kinetics of finite dose absorption through skin 2: volatile compounds. AB - A diffusion model to account for the disposition of an arbitrary dose of a (potentially) volatile compound applied to skin from a volatile vehicle is presented. In its most general form, the model allows for variable diffusivity of the permeant in the stratum corneum (SC) and must be solved numerically. However, for permeants having a constant diffusivity, absorption, and evaporation is characterized in terms of four dimensionless parameters-a reduced time tau, a fractional deposition depth in the SC f, a ratio of membrane capacity for the permeant to the applied dose beta, and a ratio of evaporative mass transfer coefficient to diffusive permeability chi. An important combination of these parameters arises as the reduced dose M(r) = (fbeta)(-1). Two cases are distinguished. In Case 1, corresponding to M(r) < or = 1, the dose is less than that required to saturate the upper layers of the SC, and the shape of the absorption and evaporation profiles is independent of the dose. Analytical solutions to Case 1 may be derived for arbitrary initial distributions of the permeant; the solution for a square wave is presented. In Case 2, corresponding to M(r) > 1, absorption and evaporation approach steady-state values as the dose is increased. Numerical evaluations of this behavior are shown. Limiting behavior for the case of a highly volatile solvent applied to skin is discussed. A companion paper discusses the application of the model to the absorption and evaporation of benzyl alcohol from human skin in vitro. PMID- 16381027 TI - Trace conditioning in 1-day-old rats. AB - A recent test of 3-hr-old rats indicated surprisingly effective trace conditioning with a 60-s trace interval. The present study tested similar trace conditioning in pups 24-hr-old, in the absence of circumstances that immediately follow birth and might promote cognition. In Experiment 1 pairing an olfactory CS with a gustatory US yielded conditioning despite a 120-s trace. Experiment 2 determined that ambient CS odor remaining in the vicinity of the conditioning surface was not sufficient to result in the olfactory-gustatory association. Experiment 3 applied conditioning procedures more like those previously used for the 3-hr-old infant and more stringent than those applied in Experiment 1. With these procedures the 24-hr-old pups nevertheless acquired the CS-US contingency with a 60-s trace. Perhaps perinatal tolerance for long trace intervals is linked to their substantial experience in dealing with odors and tastes prenatally and postnatally. PMID- 16381028 TI - Serial reversal learning of position discrimination in developing rats. AB - The current study established a procedure to evaluate the capability of rats on postnatal days (PND) 21, 26, and 30 to perform a spatial serial reversal task using a T-maze. Training consisted of an acquisition session followed by a series of six reversal sessions. To examine the role of proactive interference in the serial reversal effect, the point of reversal was manipulated so that it occurred at the start of each session (between-sessions) or the midpoint of each session (within-sessions). Performance was initially impaired during the first reversal but improved dramatically across the series. Reversal between-sessions enhanced this serial reversal effect in comparison to reversal within-sessions. Experiment 1 showed that rats of all ages learned the between-sessions serial reversal task at a comparable rate. However, on the within-sessions task, PND21 rats were impaired relative to the PND26 and 30 rats, which did not differ. Experiment 2 revealed that the addition of a tactile cue that is correlated with each phase of reversal eliminated age and task differences in serial reversal performance. These findings suggest that higher-order cognitive processes underlying serial reversal are present during the weanling period, but there is some improvement with age under conditions involving high memory interference and/or difficulty in detecting the transition between reversal phases. PMID- 16381029 TI - Spontaneous facial motility in infancy: a 3D kinematic analysis. AB - Early spontaneous orofacial movements have rarely been studied experimentally, though the motor experiences gained from these behaviors may influence the development of motor skills emerging for speech. This investigation quantitatively describes developmental changes in silent, spontaneous lip and jaw movements from 1 to 12 months of age using optically based 3D motion capture technology. Twenty-nine typically developing infants at five ages (1, 5, 7, 9, and 12 months) were studied cross-sectionally. Infants exhibited spontaneous facial movements at all ages studied. Several age-related changes were detected in lip and jaw kinematics: the occurrence of spontaneous movements increased, movement speed increased, the duration of movement epochs decreased and movement coupling among different facial regions increased. Additionally, evidence for stereotypic movements was not strong. The present findings suggest that, during the first year of life, early spontaneous facial movements undergo significant developmental change in the direction of skill development for speech. PMID- 16381031 TI - Embryonic origin of mate choice in a lizard with temperature-dependent sex determination. AB - Individual differences in the adult sexual behavior of vertebrates are rooted in the fetal environment. In the leopard gecko (Eublepharis macularius), a species with temperature-dependent sex determination (TSD), hatchling sex ratios differ between incubation temperatures, as does sexuality in same-sex animals. This variation can primarily be ascribed to the temperature having direct organizing actions on the brain. Here we demonstrate that embryonic temperature can affect adult mate choice in the leopard gecko. Given the simultaneous choice between two females from different incubation temperatures (30.0 and 34.0 degrees C), males from one incubation temperature (30.0 degrees C) preferred the female from 34.0 degrees C, while males from another incubation temperature (32.5 degrees C) preferred the female from 30.0 degrees C. We suggest that this difference in mate choice is due to an environmental influence on brain development leading to differential perception of opposite-sex individuals. This previously unrecognized modulator of adult mate choice lends further support to the view that mate choice is best understood in the context of an individual's entire life-history. Thus, sexual selection results from a combination of the female's as well as the male's life history. Female attractiveness and male choice therefore are complementary. PMID- 16381030 TI - Examining the role of endogenous opioids in learned odor-stroke associations in infant rats. AB - Maternal touch profoundly regulates infant neural and behavioral development, and supports learned odor associations necessary for infant attachment. Endogenous opioids are well characterized to mediate the calming and analgesic properties of maternal touch; yet their role in learned odor-touch associations is unknown. We administered naltrexone, an opioid receptor antagonist, before or immediately following classical conditioning with peppermint odor and tactile stimulation (stroking) in rat neonates. Results indicate odor-stroke conditioning produces odor preferences facilitated by endogenous opioids during acquisition and memory consolidation. These results provide additional evidence for the modulatory role of opioids in neonate learning and memory. Disturbances to this system may alter the impact of touch on infant development, particularly in the realm of learning necessary for attachment. PMID- 16381032 TI - Age-related changes in memory reactivation by 1- and 2-year-old human infants. AB - In three experiments, we examined the effect of a single reactivation treatment on retention by 1- and 2-year-old human infants who were tested in the visual recognition memory (VRM) paradigm. In all experiments, infants were familiarized with a visual stimulus and were tested after a delay. In the absence of a reactivation treatment, infants of both ages exhibited forgetting but exposure to a reactivation treatment alleviated forgetting after the same delay. When the duration of the original familiarization period was only 10 s, the minimum duration of an effective reminder treatment was 1 s for 2-year-olds, but was 5 s for 1-year-olds. When the duration of the original familiarization period was increased to 30 s, however, a 1-s reminder also alleviated forgetting by 1-year olds. PMID- 16381034 TI - Maternal behavior and maternal stress are associated with infant behavioral development in macaques. AB - The simultaneous effects of naturally occurring individual differences in maternal care and maternal peripartum stress on infant development have been sparsely reported in nonhuman primates. In this work, we used a comparative approach to assess how changes in peripartum maternal excreted cortisol levels and the quality of mother-infant interactions correlate with infant behavioral development in group-living rhesus and Japanese macaques. We tested the hypothesis that peripartum maternal stress was associated with infant behavioral characteristics during development. Due to the difference in mothering style between the two species, we provided separated analyses for two groups. A sample of mother-infant pairs (Japanese macaques, N = 14; rhesus macaques, N = 10) was observed during the first 3 months of the infant's life. Follow-up observations (at 5, 7, and 9 months of age) were collected for the infants. Maternal cortisol levels were measured during the peripartum period. We found preliminary evidence that maternal peripartum stress and differences in key components of maternal behavior are associated with infant behavior throughout the developmental phase. We also provided a working hypothesis regarding maternal behavior and maternal stress as factors playing unique roles in different components of infant behavioral development. PMID- 16381035 TI - Changes in the pattern of exploratory behavior are associated with the emergence of social dominance relationships in male rats. AB - This study examined the effect of the establishment of dominance relationships and subordination on exploratory behavior for both postpubertal and adult male rats. Prior to an open field test, subjects were housed either in isolation (IS) or in littermate pairs (PS) with mild dominance relationships without overt victory or defeat, or in pairs with clear hierarchical relationships as dominants (DOM) or subordinates (SUB). Stretch-attend postures and entries into the center area of the open-field were measured as an index of passive and active exploratory behavior, respectively, and crossings in the peripheral area were counted as activity. SUB rats, both postpubertal and adult, displayed less activity and lower levels of active exploratory behavior, whereas adult IS rats showed higher levels of active exploratory behavior compared to the other groups. Furthermore, both DOM and PS rats exhibited a more passive pattern of exploratory behavior in adulthood than in postpuberty. Thus the results show that an increase in the active exploratory pattern is inhibited by the establishment of social relationships among adult rats, while a decrease in activity is a primarily effect of subordination. The capacity to change exploratory patterns following subordination is found even in the postpubertal stage when adultlike social relationships have not yet appeared. PMID- 16381036 TI - Interaction of OGG1 Ser326Cys polymorphism with cigarette smoking in head and neck squamous cell carcinoma. AB - Recent molecular epidemiological studies have demonstrated that the human oxoguanine glycosylase 1 (OGG1) gene polymorphism may be associated with various cancers. To determine whether the OGG1 Ser326Cys polymorphism interacts with clinicopathological parameters including smoking and alcohol intake in head and neck squamous cell carcinoma (HNSCC), DNA samples from 192 patients with primary HNSCC were genotyped and studied by the case-only design. We observed an association between the Cys/Cys genotype and HNSCC with cigarette smoking of more than 40 pack-years by a multivariate logistic regression analysis (OR=8.10, 95% CI=1.06-61.73). No significant association of this genotype with alcohol intake was observed. Our present data suggest a possible interaction between the OGG1 Ser326Cys polymorphism and smoking in HNSCC. PMID- 16381038 TI - Cleavage of nitrogen-hydrogen bonds of ammonia induced by triruthenium polyhydrido clusters. PMID- 16381039 TI - Molecular architecture towards helical double-stranded polymers. PMID- 16381040 TI - Tribenzosubporphines: synthesis and characterization. PMID- 16381041 TI - Ruthenium half-sandwich complexes bound to protein kinase Pim-1. PMID- 16381042 TI - Phosphorus remains exciting! PMID- 16381043 TI - Total synthesis of (+/-)-merrilactone A. PMID- 16381044 TI - Biosynthesis of the cyclotide Kalata B1 by using protein splicing. PMID- 16381045 TI - Altering the regioselectivity of cytochrome P450 CYP102A3 of Bacillus subtilis by using a new versatile assay system. AB - A novel monooxygenase (CYP102A3) has been discovered within the Bacillus subtilis genome that reveals a similarity of 76 % to the well-known cytochrome P450 BM-3 of B. megaterium (CYP102A1). Both enzymes are natural fusion proteins consisting of a heme domain and a FAD/FMN-reductase domain. Because of their high turnover rates, these biocatalysts are of special interest for industrial applications, but show only limited regioselectivity. In this work, the regioselectivity of CYP102A3 was changed by directed evolution and protein design to hydroxylate substrates not only in different subterminal, but also to a high extent, in terminal carbon chain positions. To enable a high-throughput screening procedure, a very versatile assay was developed that is capable of discriminating between terminal and subterminal hydroxylation of carbon chains. A double mutant of CYP102A3 was obtained that produces 48 % octan-1-ol as the main product of the reaction. PMID- 16381046 TI - Production of Lewis x tetrasaccharides by metabolically engineered Escherichia coli. AB - Two tetrasaccharides carrying the trisaccharidic Lewis x motif on a GlcNAc or a Gal residue were produced on the gram-scale by high-cell-density cultures of metabolically engineered Escherichia coli strains that overexpressed the Helicobacter pylori futA gene for alpha-3 fucosyltransferase and the Neisseria meningitidis lgtB gene for beta-4 galactosyltransferase. The first compound Galbeta-4(Fucalpha-3)GlcNAcbeta-4GlcNAc was produced by glycosylation of chitinbiose, which was endogenously generated in the bacterial cytoplasm by the successive action of the rhizobial chitin-synthase NodC and the Bacillus circulans chitinase A1, whose genes were additionally expressed in the E. coli strain. The second compound, Galbeta-4(Fucalpha-3)GlcNAcbeta-3Gal, was produced from exogenously added Gal by a strain that was deficient in galactokinase activity and overexpressed the additional N. meningitidis lgtA gene for beta-3 N acetylglucosaminyltransferase. PMID- 16381047 TI - Termination-free prokaryotic protein translation by using anticodon-adjusted E. coli tRNASer as unified suppressors of the UAA/UGA/UAG stop codons. Read-through ribosome display of full-length DHFR with translated UTR as a buried spacer arm. PMID- 16381048 TI - DNA-templated synthesis in organic solvents. PMID- 16381050 TI - Visual essences, readable signs, means of construction: on the relevance of images in science. PMID- 16381049 TI - Biosynthesis of the heat-shock protein 90 inhibitor geldanamycin: new insight into the formation of the benzoquinone moiety. PMID- 16381051 TI - Reactive polymers: a versatile toolbox for the immobilization of functional molecules on TiO2 nanoparticles. PMID- 16381052 TI - Evidence for the formation and structure of palladacycles during Pd-catalyzed C-N bond formation with catalysts derived from bulky monophosphinobiaryl ligands. PMID- 16381053 TI - Exploitation of perfluorophenyl-phenyl interactions for achieving difficult macrocyclizations by using ring-closing metathesis. PMID- 16381054 TI - Distanna-closo-dodecaborate. PMID- 16381055 TI - Molecular-recognition characteristics of SAM-binding riboswitches. PMID- 16381057 TI - Escalation, group and A + B designs for dose-finding trials. AB - In this paper, rules are given on how to construct escalation, A + B, and group designs for dose-finding trials. Operational characteristics of these designs are discussed and compared via simulations. Dose-finding designs for trials with ordinal toxicity outcome are considered. PMID- 16381058 TI - Cooperative effect of factors governing molecular ion yields in desorption/ionization mass spectrometry. AB - Factors governing the molecular ion yields of amino acids and peptides have been studied using fast atom bombardment (FAB) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) in positive-ion mode. The ion yields of protonated amino acids under FAB conditions are dependent on proton affinity (PA), hydrophobicity, and aromaticity of amino acids. Both PA and hydrophobicity contribute to an increase in the ion yields, while aromaticity contributes to a decrease. In MALDI, the ion yields increase linearly with the increase of PA of amino acids with the exception of lysine. In both FAB and MALDI experiments with peptides, the presence of arginine residues is essential for producing abundant protonated peptides. In FAB, the presence of aliphatic and hydrophobic amino acids (leucine and isoleucine) increases the ion yields of protonated peptides, while some hydrophilic amino acids (aspartic acid and asparagines) decrease the ion yields. The presence of two or more arginine residues does not give higher ion yields in FAB. In MALDI, the presence of aromatic amino acids (phenylalanine and tyrosine) enhances the signals for protonated peptides. Thus, physicochemical factors of individual amino acids cooperatively affect the ion yields of protonated amino acids and peptides. These factors governing the ion yields in FAB and MALDI affect two processes, desorption and ionization, that can be considered independently. PMID- 16381059 TI - An ion-pairing liquid chromatography/tandem mass spectrometric method for the determination of ethephon residues in vegetables. AB - A rapid, sensitive and selective method has been developed for the direct determination of ethephon residues in vegetables (apple, cherry and tomato). Given the anionic character of ethephon, the use of ion-pairing liquid chromatography (LC) in combination with tandem mass spectrometry (MS/MS, triple quadrupole) allowed its direct determination in these matrices avoiding a derivatisation step and favouring the automation of the method. Samples were extracted with a mixture of dichloromethane/aqueous formic acid (pH 3) (1:1). Then, tetrabutylammonium acetate (TBA) was added as an ion-pairing reagent, and an aliquot of the aqueous extract was directly injected into the LC/MS/MS system. Quantification was performed with matrix-matched standards prepared from blank sample extracts. MS/MS measurements were made in the selected reaction monitoring (SRM) mode, using the most sensitive transition (m/z 107 > 79) for quantification, and up to four additional transitions for confirmation. Quantitative recoveries were obtained for all matrices (between 83% and 96%) at two concentration levels tested (0.05 and 0.5 mg/kg), with relative standard deviations lower than 9% in all cases. The addition of TBA directly into the sample extract contained in the injection vial was found sufficient to obtain satisfactory LC retention for the analyte. Under these conditions, the absence of ion-pairing reagent in the mobile phase minimised the ionisation suppression for ethephon in the MS source, leading to an increase in the sensitivity of the method and reaching limits of detection of 0.02 mg/kg for all matrices investigated. The acquisition of five specific MS/MS transitions for ethephon allowed the simultaneous and reliable quantification and confirmation of the analyte in the samples. PMID- 16381060 TI - Temperature dependence of the mean autodetachment lifetime of the p-benzoquinone molecular radical anion. AB - Experimental data previously published from this laboratory (Asfandiarov NL, Pshenichnyuk SA, Fokin AI, Nafikova EP. Chem. Phys. 2004; 298: 263), on the temperature dependence of the mean autodetachment lifetime of the p-benzoquinone molecular radical anion, were analyzed in the framework of a simple statistical model. This model is a simplification of the well-known semiempirical approach elaborated by Christophorou et al. (Christophorou LG, Hadjiantoniou A, Carter JG. J. Chem. Soc., Faraday Trans. II 1973; 69: 1722). Evaluation of the vibrational energy storage of the target molecule and anion was made in a quantum approximation following a published procedure (Matejcik S, Mark TD, Spanel P, Smith D, Jaffke T, Illenberger E. J. Chem. Phys. 1994; 102: 2516). The results obtained are in reasonable agreement with the experimental data. PMID- 16381061 TI - Desorption electrospray ionization mass spectrometry for the analysis of pharmaceuticals and metabolites. AB - The performance of desorption electrospray ionization (DESI) in the analysis of a group of pharmaceuticals and their glucuronic acid conjugates is reported. The suitability of different sprayer solvents and different surfaces was examined. In the positive ion mode, water/methanol/trifluoroacetic acid performed best, whereas, in the negative ion mode, water/methanol/ammonium hydroxide was found to be the most suitable spray solvent. Of the surfaces investigated, polymethylmethacrylate (PMMA) was found to give the best performance in terms of sensitivity. Spray solution flow rate and the distance of the sprayer tip from the surface were also found to have significant effects on the signal intensity. Analytes with basic groups efficiently formed the corresponding protonated molecules in the positive ion mode, whereas acidic analytes, such as the glucuronic acid conjugates, formed intense signals due to the deprotonated molecules in the negative ion mode. Ionization of neutral compounds was less efficient and in many cases it was achieved through adduct formation with simple anions or cations. PMID- 16381062 TI - Analysis of unknown compounds in gentamicin bulk samples with liquid chromatography coupled with ion trap mass spectrometry. AB - Six unknown compounds present in bulk gentamicin samples have been identified by ion-pairing reversed-phase liquid chromatography coupled with ion trap mass spectrometry. The structures of these unknown compounds were deduced by comparison of their fragmentation patterns with those of the available related substances and gentamicin reference substances. Seven other unknown components were partially identified. PMID- 16381063 TI - Screening for pregnane glycosides with immunological activities from the stems of Stephanotis mucronata by high-performance liquid chromatography/tandem mass spectrometry. AB - The stems of the Chinese traditional medicine Stephanotis mucronata were screened for immunologically active pregnane glycosides using high-performance liquid chromatography (HPLC) coupled with electrospray ionization tandem mass spectrometry. In the mass spectra of pregnane glycosides, predominant [M+Na]+ ions were observed and used to determine the molecular masses, while fragmentation reactions of the [M+Na]+ ions were recorded to provide information on the primary sequences of oligosaccharide chains in terms of classes of monosaccharide. Fragment ions from the side-chain cleavage of aglycone portions can provide mass information about side-chain substitutions. To further confirm the fragment ion structures, Fourier transform ion cyclotron resonance tandem mass spectrometry (MSn) with low-energy collision-induced dissociation was performed using samples collected from HPLC fractions, which provided accurate elemental compositions of fragment ions. Based on fragmentation patterns and comparison with standards, ten pregnane glycosides were identified as stemucronatosides C, D, F, and G, mucronatosides A, B, and C, stephanoside E, and two glycosides that are identified in the S. mucronata extracts for the first time. The latter two pregnane glycosides are 12-O-cinnamoyldeacetylmetaplexigenin 3-O-6-deoxy-3-O-methyl-beta-D-allopyranosyl-(1 --> 4)-beta-D-cymaropyranosyl-(1 - > 4)-beta-D-cymaropyranoside and 12-O-cinnamoyl-20-O-acetyl (20S)-pregn-6-ene 3beta,5alpha,8beta,12beta,14beta,17beta,20-heptaol 3-O-beta-D-thevetopyranosyl-(1 --> 4)-beta-D-cymaropyranosyl-(1 --> 4)-beta-D-cymaropyranoside. PMID- 16381064 TI - A new method of transvaginal ultrasound-guided polypectomy: a feasibility study. AB - OBJECTIVES: A new device has been manufactured (Safe T Choice), which allows attachment of a transvaginal ultrasound probe to a specially adapted cervical tenaculum. This affords the capacity to monitor intrauterine surgical procedures without the need for hysteroscopy. The purpose of this study was to investigate the feasibility of endometrial polypectomy using this device combined with saline contrast sonohysterography (SCSH) to monitor the procedure. METHODS: Women diagnosed with an endometrial polyp on routine B-mode two-dimensional transvaginal ultrasound (TVS) were invited to join the study. Transvaginal ultrasound-guided polypectomies were carried out by a single operator. The procedure was timed from application until removal of the tenaculum. The ultrasound views were rated as satisfactory or poor. Success of the procedure was gauged by complete removal of the polyp without recourse to hysteroscopy. Women also attended for postoperative follow-up ultrasound scans to check for residual disease. RESULTS: Thirty-seven women were recruited to the study. The mean operating time was 8 min (95% CI, 5.9-10.4). The procedure was successful in 32/37 (86.5%) cases (95% CI, 75.5-97.5). In three cases (8.1%) the procedure failed because of an inability to obtain satisfactory images of the uterine cavity, and in two further cases (5.4%) the operator was unable to grasp and remove the polyp. Two patients (5.4%) bled from the tenaculum insertion site, necessitating suture for hemostasis. There were no other complications and none of the patients had evidence of residual polyp tissue at the follow-up visit. CONCLUSION: This study showed that transvaginal ultrasound-guided polypectomy is a feasible technique for the removal of endometrial polyps. Further work is required to compare outcomes and cost-effectiveness of this technique with hysteroscopic polypectomy. PMID- 16381065 TI - Structural assignment of isomeric 2-aminopyridine-derivatized monosialylated biantennary N-linked oligosaccharides using negative-ion multistage tandem mass spectral matching. AB - To investigate the possibility of structural assignment based on negative-ion tandem multistage (MSn) mass spectral matching, four isomers of 2-aminopyridine (PA)-derivatized monosialylated oligosaccharides (i.e., complex-type N-glycans with an alpha2-3- or alpha2-6-linked sialic acid on alpha1-6 or alpha1-3 antennae) were analyzed using high-performance liquid chromatography/electrospray ion trap time-of-flight mass spectrometry (HPLC/ESI-IT-TOFMS). The negative ion [M-2H]2- is observed predominantly in the MS1 spectra without the loss of a sialic acid. The MS2 spectra derived from it are sufficiently reproducible that MS2 spectral matching based on correlation coefficients can be applied to the assignment of these isomers. The isomers containing a sialic acid on alpha1-6 or alpha1-3 antennae can be distinguished by MS2 spectral matching, but the alpha2-3 and alpha2-6 linkage types of sialic acid cannot be distinguished by their MS2 spectra. However, MS3 spectra derived from fragment ions containing a sialic acid (i.e., C4- and D-type ions) clearly differentiate the alpha2-3 and alpha2-6 linkage types of sialic acid in their MS3 spectral patterns. This difference might be rationalized in terms of a proton transfer from the reducing-end mannose to the negatively charged sialic acid. These two moieties are very close in the structural conformations of the precursor C4-type fragment ions of alpha2-6 linkage type, as predicted by molecular mechanics calculations. Thus, negative ion MSn (n = 2, 3) spectral matching was demonstrated to be useful for the structural assignment of these four monosialylated PA N-glycan isomers. PMID- 16381066 TI - Barriers to the provision of evidence-based psychosocial care in oncology. AB - Meeting the psychological, social and physical needs of people with cancer is a challenge for individual health practitioners, health administrators and health policy makers. However, there is a considerable gap between recommended best evidence psychosocial and supportive care and actual practice. This paper provides a discussion of the reasons for this gap using the precede-proceed model as a theoretical framework. The model is a useful way of classifying potential barriers to the application of recommended best practice into three categories: predisposing factors which influence motivation to behave in a particular way, enabling factors which facilitate the enactment of the behaviour and reinforcing factors which increase the likelihood that the behaviour will be maintained over time. Ways of addressing these barriers are proposed and discussed. PMID- 16381067 TI - Statistical inference for correlated data in ophthalmologic studies. AB - In ophthalmologic studies, each subject usually contributes important information for each of two eyes and the values from the two eyes are generally highly correlated. Previous studies showed that test procedures for binary paired data that ignore the presence of intraclass correlation could lead to inflated significance levels. Furthermore, it is possible that asymptotic versions of these procedures that take the intraclass correlation into account could also produce unacceptably high type I error rates when the sample size is small or the data structure is sparse. We propose two alternatives for these situations, namely the exact unconditional and approximate unconditional procedures. According to our simulation results, the exact procedures usually produce extremely conservative empirical type I error rates. That is, the corresponding type I error rates could greatly underestimate the pre-assigned nominal level (e.g. (empirical type I error rate/nominal type I error rate) 0.8). On the other hand, the approximate unconditional procedures usually yield empirical type I error rates close to the pre-chosen nominal level. We illustrate our methodologies with a data set from a retinal detachment study. PMID- 16381068 TI - Diazepam-induced prospective memory impairment and its relation to retrospective memory, attention, and arousal. AB - The amnestic effects of benzodiazepines are well documented on a variety of memory tasks. However, prospective memory (PM), or remembering to execute an action at a future time, has not been studied previously. This study examined the effect of diazepam on word list recall, PM, sustained attention, and subjective ratings of arousal. Forty-eight healthy participants, aged 19-35, received an average of 0.19 mg/kg oral diazepam or placebo in a double-blind manner. Retrospective memory and PM were assessed by free recall of unrelated word lists and by instructing participants to request a hidden belonging at the end of the session, respectively. Sustained attention was measured by multiple trials of a digit cancellation task, and subjective arousal was assessed by self-ratings of drowsiness. Diazepam impaired performance on all measures, including PM. Reduced PM performance was associated with decreased subjective arousal in the diazepam group but was unrelated to sustained attention. This is the first report of the effects of benzodiazepines on prospective remembering, and further supports the view that the arousal/attentional system is composed of partially independent subsystems that have differential relationships to memory. PMID- 16381069 TI - The sigmoidally transformed cosine curve: a mathematical model for circadian rhythms with symmetric non-sinusoidal shapes. AB - We introduce a family of non-linear transformations of the traditional cosine curve used in the modelling of biological rhythms. The non-linear transformation is the sigmoidal family, represented here by three family members: the Hill function, the anti-logistic function, and the arctangent function. These transforms add two additional parameters that must be estimated, in addition to the acrophase, MESOR, and amplitude (and period in some applications), but the estimated curves have shapes requiring many more than two additional harmonics to achieve the same fit when modelled by harmonic regression. Particular values of the additional parameters can yield rectangular waves, narrow pulses, wide pulses, and for rectangular waves (representing alternating 'on' and 'off' states) the times of onset and offset (hence duration, as when modelling the duration of the large night-time melatonin secretory epoch). We illustrate the sigmoidally transformed cosine curves, and compare them to harmonic regression modelling, in a sample of eight activity recordings made on patients in a nursing home. PMID- 16381070 TI - Non-inferiority trials: the 'at least as good as' criterion with dichotomous data. AB - The 'at least as good as' criterion, introduced by Laster and Johnson for a continuous response variate, is developed here for applications with dichotomous data. This approach is adaptive in nature, as the margin of non-inferiority is not taken as a fixed difference; it varies as a function of the positive control response. When the non-inferiority margin is referenced as a high fraction of the positive control response, the procedure is seen to be uniformly more efficient than the fixed margin approach, yielding smaller sample sizes when sizing non inferiority trials under identically specified conditions. Extending this method to proportions is straightforward, but highlights special considerations in the design of non-inferiority trials versus superiority trials, including potential trade-offs in statistical efficiency and interpretability. PMID- 16381071 TI - Bivariate poisson-poisson model of zero-inflated absenteeism data. AB - Bimodal distributions of counts with one mode at zero are often seen in medical research. In a health survey parents were asked the number of days their children missed their activities (Y(1)) and the number of days their children spent in bed (Y(2)) due to illness in the past four weeks. Both variables exhibited zero inflation. We consider a bivariate Poisson-Poisson regression model, in which the two variables are regarded as indicators of an unobserved health status variable. Based on this, we further develop a bivariate Poisson-Poisson model that constrains Y(1)>or=Y(2). It is often claimed that there is a critical window of growth and nutrition in foetal life and infancy during which subsequent health status is affected. It is not clear whether the claim is true and whether childhood growth matters more. We analyse the bivariate data in relation to weight-for-age in infancy and weight gain from infancy to age 7 years. The findings do not support the existence of a critical window in infancy. There is some indication that childhood weight gain might affect health status. PMID- 16381072 TI - Extending the methods used to screen the WHO drug safety database towards analysis of complex associations and improved accuracy for rare events. AB - Post-marketing drug safety data sets are often massive, and entail problems with heterogeneity and selection bias. Nevertheless, quantitative methods have proven a very useful aid to help clinical experts in screening for previously unknown associations in these data sets. The WHO international drug safety database is the world's largest data set of its kind with over three million reports on suspected adverse drug reaction incidents. Since 1998, an exploratory data analysis method has been in routine use to screen for quantitative associations in this data set. This method was originally based on large sample approximations and limited to pairwise associations, but in this article we propose more accurate credibility interval estimates and extend the method to allow for the analysis of more complex quantitative associations. The accuracy of the proposed credibility intervals is evaluated through comparison to precise Monte Carlo simulations. In addition, we propose a Mantel-Haenszel-type adjustment to control for suspected confounders. PMID- 16381073 TI - Comparison of misclassification rates of search partition analysis and other classification methods. AB - Search partition analysis (SPAN) is a method to develop classification rules based on Boolean expressions. The performance of SPAN is compared against the trials reported by Lim et al. of 33 other methods of classification, including tree, neural network and regression methods on 16 data sets, most of which were health related. Each data set was augmented with noise variables in further trials. Lim et al. assessed the performance of the methods by estimates of misclassification rate, either cross-validated or test sample based. In this paper, the same data sets are analysed by SPAN and misclassification rates of the SPAN classifiers are estimated. Comparison is made of the performance of SPAN against the other methods that were considered by Lim et al. In terms of average misclassification error rate, taken over all data sets, SPAN was among the best five methods. In terms of average ranking of misclassification, that is, for each data set ranking the misclassification rates from lowest to highest, SPAN was second only to polyclass logistic regression. PMID- 16381074 TI - Examining accuracy of screening mammography using an event order model. AB - Screening mammography is a widely used method for breast cancer detection. For each mammogram we propose a performance model based on order of outcomes. That is, we envision an initial assessment, a follow up assessment if the initial one is positive and, eventually, a determination of whether cancer was present or not. A model can be built at each stage reflecting effects due to patient characteristics, to the facility where mammogram was performed and to the radiologist reading the mammogram. Since assessment is not perfectly associated with outcome, familiar rates of agreement and disagreement are of interest. These rates can be investigated at various levels of risk factors of interest. The approach is illustrated with screening mammography data from the Group Health Cooperative in Seattle, WA. A Bayesian framework is adopted for inference and an analysis of the data set is presented. PMID- 16381075 TI - A Bayesian semi-parametric model for colorectal cancer incidences. AB - A Bayesian semi-parametric model is proposed to capture the interaction among demographic effects (age and gender), spatial effects (county) and temporal effects of colorectal cancer incidences simultaneously. In particular, an extension of multivariate conditionally autoregressive (CAR) processes to a partially informative Gaussian demographic spatial temporal CAR (DSTCAR) process for a spatial-temporal setting is proposed. The precision matrix of the Gaussian DSTCAR process is the Kronecker product of several components. The spatial component is modelled with a CAR prior. A pth order intrinsic autoregressive prior (IAR(p)) is implemented for the temporal component to estimate a smoothed and non-parametric temporal trend. The demographic component is modelled with a Wishart prior. Data analysis shows significant spatial correlation only exists in the age group of 50-59. Males and females in their 50s and 60s show fairly strong correlation. The hypothesis testing based on Bayes factor suggests that gender correlation cannot be ignored in this model. PMID- 16381076 TI - Surrogate markers: back to the future. PMID- 16381077 TI - Phase specific analysis of herpes zoster associated pain data: a new statistical approach by R. B. Arani, S.-J. Soong, H. L. Weiss, M. J. Wood, P. A. Fiddian, J. W. Gnann and R. Whitley, Statistics in Medicine 2001; 20:2429-2439. PMID- 16381078 TI - Optimized and parallelized implementation of the electronegativity equalization method and the atom-bond electronegativity equalization method. AB - The most common way to calculate charge distribution in a molecule is ab initio quantum mechanics (QM). Some faster alternatives to QM have also been developed, the so-called "equalization methods" EEM and ABEEM, which are based on DFT. We have implemented and optimized the EEM and ABEEM methods and created the EEM SOLVER and ABEEM SOLVER programs. It has been found that the most time-consuming part of equalization methods is the reduction of the matrix belonging to the equation system generated by the method. Therefore, for both methods this part was replaced by the parallel algorithm WIRS and implemented within the PVM environment. The parallelized versions of the programs EEM SOLVER and ABEEM SOLVER showed promising results, especially on a single computer with several processors (compact PVM). The implemented programs are available through the Web page http://ncbr.chemi.muni.cz/~n19n/eem_abeem. PMID- 16381079 TI - Multibaric-multithermal ensemble molecular dynamics simulations. AB - We present new generalized-ensemble molecular dynamics simulation algorithms, which we refer to as the multibaric-multithermal molecular dynamics. We describe three algorithms based on (1) the Nose thermostat and the Andersen barostat, (2) the Nose-Poincare thermostat and the Andersen barostat, and (3) the Gaussian thermostat and the Andersen barostat. The multibaric-multithermal simulations perform random walks widely both in the potential-energy space and in the volume space. Therefore, one can calculate isobaric-isothermal ensemble averages in wide ranges of temperature and pressure from only one simulation run. We test the effectiveness of the multibaric-multithermal algorithm by applying it to a Lennard-Jones 12-6 potential system. PMID- 16381080 TI - Constructing binomial confidence intervals with near nominal coverage by adding a single imaginary failure or success. AB - In this paper we present a simple method for constructing (1- alpha)100 per cent confidence intervals for binomial proportions with near nominal coverage for all underlying proportion parameters on the unit interval. This new method uses, with a slight modification, the standard normal approximation technique taught in introductory statistics classes. Specifically, we first augment the observed binomial data with an imaginary failure to compute the lower bound and an imaginary success to compute the upper bound. By contrast, the Agresti-Coull method adds the same number of imaginary successes and failures to the observed data, yet it can still give somewhat subnominal coverage. As motivation, we discuss the relationship between this new method and the Clopper-Pearson exact method. We also present numerical calculations to illustrate the satisfactory performance of this new method compared with several common alternatives. Finally, we argue that for certain statistical applications, such as the design and analysis of clinical trials with adverse events, this new method represents a valuable complementary approach. PMID- 16381081 TI - Road traffic injuries in the elderly. AB - OBJECTIVE: Elderly victims of motor vehicle collisions are increasing with the aging population. This study aimed to investigate the injury pattern of elderly victims involved in motor vehicle collisions. METHODS: This was a retrospective study using data from the Victorian State Trauma Outcome Registry and Monitoring Group (VSTORM) from June 2001 to July 2003, Australian Bureau of Statistics year 2001 population estimates, and Victoria Transport Accident Commission year 2001 total road death toll. Elderly victims were defined as age 65 and above. Comparison of fatality rates and general injury patterns for the elderly and young victims was undertaken. RESULTS: The total fatality rate of the elderly group was almost double that of the younger group. The elderly victims had a higher rate of chest injuries (23.42% v 18.17%; p = 0.003). The three most common chest injuries of the elderly victims were rib fractures (23.58%), flail chest (9.55%), and sternum fractures (5.97%). Elderly chest injured patients also had longer intensive care unit stay compared with the younger group (7.96 days v 5.31 days; p = 0.048). CONCLUSIONS: Elderly victims of motor vehicle collisions have a higher risk of chest injuries, especially of chest wall injuries. Age specific injury patterns are important in determining primary and secondary prevention strategies. PMID- 16381082 TI - Using simulation to estimate the cost effectiveness of improving ambulance and thrombolysis response times after myocardial infarction. AB - OBJECTIVES: To quantify the health gains and costs associated with improving ambulance and thrombolysis response times for acute myocardial infarction. DESIGN: A computer simulation model. PATIENTS/SETTINGS: Patients experiencing acute myocardial infarction in England. INTERVENTIONS: Improving the ambulance response time to 75% of calls reached within 8 minutes and the hospital arrival to thrombolysis time interval (door-to-needle time) to 75% receiving it within 30 minutes and 20 minutes, compared to best estimates of response times in the mid 1990s. MAIN OUTCOME MEASURES: Deaths prevented, life years saved, and discounted cost per life year saved. RESULTS: Improving the ambulance response to 75% of calls within 8 minutes resulted in an estimate of 5 deaths prevented or 57 life years saved per million population per year, with a discounted incremental cost per life year saved of 8540 pounds sterling over 20 years. The corresponding benefit of improving the door-to-needle time to 75% of myocardial infarction patients within 30 minutes was an estimated 2 deaths prevented and 15 life years saved per million population per year, with a discounted incremental cost per life year saved of between 10,150 pounds sterling to 54,230 pounds sterling over 20 years. Little further gain was associated with reaching the 20 minute target. Combining ambulance and thrombolysis targets resulted in 70 life years saved per million population per year. CONCLUSIONS: Improving ambulance response times appears to be cost effective. Reducing door-to-needle time will have a smaller effect at an uncertain cost. Further benefits may be gained from reducing the time from onset of symptoms to starting thrombolysis. PMID- 16381083 TI - Tinidazole: a new super-charged metronidazole. PMID- 16381084 TI - Rhabdomyolysis. PMID- 16381085 TI - Biomarkers for preterm birth. PMID- 16381086 TI - Plasma exposure alters the proteome of S. pyogenes. PMID- 16381087 TI - Proteomics gets out of a fix. PMID- 16381088 TI - From clinical research to clinical practice: a 4-year review of ziprasidone. AB - Ziprasidone is a second-generation antipsychotic that received Food and Drug Administration approval in February 2001. It has a unique receptor profile that includes high-affinity antagonist activity at dopamine D2 receptors, inverse agonist activity at serotonin (5-HT)2A receptors, agonist activity at 5-HTlA receptors, and a relatively high affinity for the serotonin and norepinephrine transporters. The 5-HTIA affinity, together with the inhibitory effect on mono amine reuptake, may underlie the hypothesized beneficial effects on comorbid affective and cognitive abnormalities in schizophrenia and schizoaffective disorder. The short-term efficacy of ziprasidone for core positive symptoms of schizophrenia appears to be comparable to other conventional and atypical antipsychotics. The short-term efficacy of ziprasidone in acute mania has been established based on two 3-week, double-blind, placebo-controlled trials.Open label treatment for up to 52 weeks confirms the sustained efficacy and safety of ziprasidone in bipolar disorder. Maintenance studies in schizophrenia and schizoaffective disorder indicate that long-term ziprasidone therapy is effective in preventing relapse, while maintaining cognitive and psychosocial benefits. The safety database suggests that the overall cardiovascular and cerebrovascular risk associated with ziprasidone is lower than with other atypicals, with notably lower risk of drug-related increases in weight, glucose, or lipids. The data also suggest a modestly increased risk of QTc prolongation that is not dose related or linked to torsades de pointes. Switching to ziprasidone from other atypicals appears to improve both clinical symptoms and metabolic parameters, though more studies are needed to fully characterize these benefits. This monograph summarizes the efficacy, tolerability, and safety of oral ziprasidone in the treatment of schizophrenia, schizoaffective disorder, and bipolar mania. PMID- 16381089 TI - Response times: myths, measurement & management. PMID- 16381090 TI - The education bridge: how culture-based narratives improve EMS learning. PMID- 16381091 TI - A two-way street: Kansas adopts bi-directional education mobility for registered nurses and paramedics. PMID- 16381092 TI - Days of triage: familiarity breeds controlled chaos. PMID- 16381093 TI - Spinal trauma: prehospital identification & management of spinal-injured patients. PMID- 16381094 TI - Screening for duct-dependant congenital heart disease with pulse oximetry: a critical evaluation of strategies to maximize sensitivity. AB - AIM: To evaluate the feasibility of detecting duct-dependent congenital heart disease before hospital discharge by using pulse oximetry. DESIGN: Case-control study. SETTING: A supra-regional referral centre for paediatric cardiac surgery in Sweden. PATIENTS: 200 normal term newborns with echocardiographically normal hearts (median age 1.0 d) and 66 infants with critical congenital heart disease (CCHD; median age 3 d). METHODS: Pulse oximetry was performed in the right hand and one foot using a new-generation pulse oximeter (NGoxi) and a conventional technology oximeter (CToxi). RESULTS: With the NGoxi, normal newborns showed a median postductal saturation of 99% (range 94-100%); intra-observer variability showed a mean difference of 0% (SD 1.3%), and inter-observer variability was 0% (SD 1.5%). The CToxi recorded a significantly greater proportion of postductal values below 95% (41% vs 1%) in the normal newborns compared with NGoxi (p<0.0001). The CCHD group showed a median postductal saturation of 90% (45-99%) with the NGoxi. Analysis of distributions suggested a screening cut-off of <95%; however, this still gave 7/66 false-negative patients, all with aortic arch obstruction. Best sensitivity was obtained by adding one further criterion: saturation of <95% in both hand and foot or a difference of >+/-3% between hand and foot. These combined criteria gave a sensitivity of 98.5%, specificity of 96.0%, positive predictive value of 89.0% and negative predictive value of 99.5%. CONCLUSION: Systematic screening for CCHD with high accuracy requires a new generation oximeter, and comparison of saturation values from the right hand and one foot substantially improves the detection of CCHD. PMID- 16381095 TI - A renaissance for the 'sense of wonder'? PMID- 16381096 TI - What price dissection? Dissection literally dissected. AB - Hamlet: Has this fellow no feelings of his business, that he sings at grave making? Horatio: Custom hath made it in him a property of easiness. (Hamlet Act V, scene i) Hamlet is appalled by the gravedigger's insensitivity towards death and corpses. Horatio explains that the gravedigger is so accustomed to such things that he no longer shares Hamlet's seriousness. We contend that human dissection may make in medical students and doctors the "property of easiness" in dealing with death and the human body, and that this may have negative consequences for medics and patients. It is perhaps worth emphasising at the outset what this essay is NOT about. We do not wish to call into question the value of dissection in medical education; to charge dissection with being an inefficient or ineffective means of teaching and learning human anatomy is not our intent. Instead, we explore, through the medium of literature, experiences of dissection, and what kind of student and doctor may be encouraged or produced by the dissection room; what price might be paid for a practical, first-hand experience of human anatomy. PMID- 16381097 TI - [Prevalence of serum antibodies against Bartonella ssp. in a health population from the south area of the Seville province]. AB - OBJECTIVE: To know the prevalence of serum antibodies against Bartonella spp. in a healthy population from south of Spain. PATIENTS AND METHODS: A clinical epidemiological survey was conducted among 146 healthy individuals. An indirect immunofluorescence commercial technique was used in a sample of serum from each individual to detect the present of IgG type serum antibodies against Bartonella spp., considering a result equal to or greater than 1:128 as positive. RESULTS: Thirty six (24.7%) of all the subjects studied were asymptomatic carriers of antibodies against Bartonella spp. No crossed reactions against Chlamydia trachomatis, C. pneumoniae or Coxiella burnetti were observed. No significant association was found between the presence of seropositivity for Bartonella spp. and other factors. CONCLUSION: There is an elevated frequency of asymptomatic carriers of antibodies against Bartonella spp. among the healthy population of our area. This suggests that most of the infections by Bartonella are subclinical. PMID- 16381098 TI - The medical ethics of Erasmus and the physician-patient relationship. AB - Desiderius Erasmus set out his views on medical ethics just over 500 years ago. Applying the characteristic approach of Renaissance Humanism, he drew upon a variety of classical sources to develop his own account of medical obligation. Of particular interest is Erasmus's attention to the patient's duties as well as the physician's. By treating this reciprocal relationship as a friendship between extreme unequals, Erasmus was able to maintain the nobility of the medical art and at the same time deal with the culturally sensitive issue of payment for physicians' services. The use of physician-patient reciprocity as a principle of medical ethics has until recently been considered a novel feature of nineteenth century medical codes. As Erasmus's treatment of physician-patient reciprocity arose from a classical conception of friendship, there may be grounds for reconsidering the role of friendship in other discourses on medical ethics from the Renaissance to the nineteenth century. PMID- 16381099 TI - Medical humanities at the University of Wales Swansea. AB - The UK's first taught master's degree in medical humanities involves a field of inquiry that is frequently philosophical, pursuing interests and questions traditionally arising in medical philosophy and ethics, but on a larger interdisciplinary canvas, drawing upon literature and the visual arts, sociology and anthropology, social history and politics, and theological and religious perspectives. PMID- 16381100 TI - [Do you see morning red in the east?]. PMID- 16381101 TI - [The ophthalmologist on the Internet]. PMID- 16381102 TI - From interleukin families to glycans: relating cytokine structure to function. PMID- 16381103 TI - Post-traumatic stress disorder. PMID- 16381104 TI - Edna Mabel Auger (1876-1932). AB - In June 1924, the full-time instructress at Medicine Hat General Hospital (MHGH) school of nursing and all the graduated nurses on staff at the hospital resigned (MHGH Minutes of the Board, 1924). Because of cutbacks, the school's superintendent, Edna Auger, was unable to hire an instructress again until 1928, and so she took on most of the teaching duties herself (MHGH Minutes of the Board, 1928). The cause of the group resignation remains clouded in controversy. One theory holds that a book written by Auger and approved by the hospital board in 1922, Rules and Regulations for Nurses, caused some dissension among staff members and their ultimate resignations. PMID- 16381105 TI - White uniforms... to be or not to be, that is the question. PMID- 16381108 TI - The "drip-and-ship" approach: starting IV t-PA for acute ischemic stroke at outside hospitals prior to transfer to a regional stroke center. AB - PURPOSE: Acute stroke therapy with intravenous (IV) tissue plasminogen activator (t-PA) is vastly underutilized. Increasingly, patients are being started on IV t PA and being transferred to regional Stroke Center programs, where additional therapies can be offered in a multimodal format. We describe our experience at the Stroke Center at Hartford Hospital with interhospital patient transfers who received IV t-PA prior to transfer to our medical center. METHODS: A retrospective analysis of our Acute Stroke Therapies database was undertaken, encompassing the intial four-year period of our Stroke Center program (May 1, 2001 to April 30, 2005). We evaluated the patient characteristics, clinical outcomes, and adjunctive therapies of patients who were started on IV t-PA at referring hospitals prior to their emergent transfer to our Stroke Center. RESULTS: From a total of 229 patients who received IV and/or IA thrombolysis and newer catheter-delivered devices or clinical trials at our Stroke Center, a total of 33 (14.4%) were started on IV t-PA at an outside hospital prior to transfer. Symptomatic hemorrhage occurred in one of the 33 patients (3.0%), and in-hospital mortality rate for these patients was 6.1%. A total of 26 patients (78.8%) had a positive outcome in that they were discharged either to home or to acute rehabilitation. CONCLUSIONS: Use of IV t-PA in a "drip-and-ship" approach is growing at the regional Stroke Center at Hartford Hospital. This protocol is safe and offers several advances for the care of patients with AIS: (a) empowering emergency physicians and neurologists at outside hospitals, via access to a 24/7 Acute Stroke Hotline, to treat patients with AIS; (b) facilitating the early initiation of IV t-PA; and (c) offering adjunctive therapeutic approaches, following arrival at our facility, for patients not sufficiently improving with IV t-PA alone. PMID- 16381109 TI - Discharge patterns of patients with asthma from the emergency department: a retrospective review. AB - BACKGROUND: Despite evidence that specific therapies improve outcomes in patients with asthma, they are often not used. Combining several evidence-based therapies into a treatment "bundle" to be offered at the time of discharge from the emergency department, might reduce variation and potentially optimize clinical outcomes. OBJECTIVE: To assess the utilization of four evidence-based therapies for asthma by analyzing the visits of patients with acute exacerbations of asthma discharged from the emergency department. DESIGN: A retrospective chart review. SETTING: Single 650-bed inner-city hospital emergency department. PATIENTS: Two hundred and twenty six patients discharged from the emergency department after 500 acute exacerbations of asthma. MEASUREMENTS: All visits were reviewed for the presence of the four evidence-based components of asthma treatment upon discharge: follow-up referral, oral steroids, asthma education, and inhaled corticosteroids. Visits were also assessed for medications prescribed upon discharge, medication history, and patient's asthma severity based on national guidelines. RESULTS: The four components of asthma treatment were documented as follows: follow-up referral (86.2%), oral steroids (67.8%), asthma education (19.6%), and inhaled corticosteroids (16.2%). Only 3.4% of visits documented all four components in the aggregate. Twenty-three distinct combinations of medication were prescribed upon discharge. The majority of visits failed to document asthma severity. CONCLUSIONS: This retrospective chart review reveals significant variation in the discharge management of patients with asthma, specifically regarding medications prescribed. While follow-up referral was sufficiently documented, the remaining three components were not. With only 3.4% of visits containing all four components, implementing an asthma "bundle" may present an opportunity to improve outcomes in asthma management. PMID- 16381110 TI - Malignant meningioma: computed tomography and magnetic resonance imaging characteristics. AB - Malignant meningioma is a relatively rare subtype of meningioma demonstrating an aggressive growth pattern, distinct histologic features, and a propensity for recurrence following surgical resection. There has been little consensus regarding imaging features of malignant meningioma, and typically the degree of tumor aggressiveness is determined by pathology. Here we describe a case of malignant meningioma determined to be benign at initial biopsy. Computed tomography (CT) and magnetic resonance (MR) imaging are reviewed with discussion of the aggressive features seen at presentation. PMID- 16381111 TI - A liver transplant: the gift of life. PMID- 16381112 TI - Are public relations necessary to a medical society? 1955. PMID- 16381113 TI - Work: satisfactions and dissatisfactions. PMID- 16381114 TI - Antibiotics and mental disorder. PMID- 16381115 TI - What you know, what you have, what you are. PMID- 16381116 TI - Arming labs with biopreparedness tools. PMID- 16381117 TI - Patient monitoring in the Fast Lane. PMID- 16381118 TI - Embedded coaches lead to CPOE victory. PMID- 16381119 TI - Formulary for success. PMID- 16381120 TI - Raising the bar. PMID- 16381121 TI - A dose of RHIOlity. PMID- 16381122 TI - Medical simulation makes medical education better and safer. PMID- 16381123 TI - Voice medicine: arts medicine for otolaryngologists. PMID- 16381124 TI - Blast injury to the tympanic membrane. PMID- 16381125 TI - Endoscopic view of obstructing frontal recess polyps. PMID- 16381126 TI - Mucus retention cyst and reflux disease. PMID- 16381127 TI - Impacted pharyngeal fish bone migrating to the retropharynx. PMID- 16381128 TI - Osteomyelitis. PMID- 16381129 TI - Dedifferentiated cervical liposarcoma. PMID- 16381130 TI - The lower-eyelid tarsal-strip procedure. PMID- 16381131 TI - Class action lawsuits. PMID- 16381132 TI - Spontaneous transtemporal CSF leakage: a study of 51 cases. AB - We conducted a retrospective study of 51 cases of spontaneous transtemporal cerebrospinal fluid (CSF) leakage in 48 adults who had presented to our tertiary care academic referral center between July 1, 1988, and June 30, 2002. All patients had undergone high-resolution temporal bone computed tomography, and 26 patients had undergone magnetic resonance imaging. All patients were treated with a middle fossa craniotomy to repair the CSF fistulae. During a mean follow-up of 4.9 years, 46 of the 48 patients (95.8%) had experienced a complete cessation of CSF leakage (49 of 51 cases [96.1%]). The 2 patients whose leakage recurred were successfully managed with a subtotal petrosectomy with occlusion of the eustachian tube and obliteration of the middle ear and mastoid. No patient developed meningitis. PMID- 16381133 TI - Poststapedectomy hearing gain: comparison of a Teflon (fluoroplastic ASTM F 754) prosthesis with a Schuknecht-type wire/Teflon prosthesis. AB - We conducted a retrospective study to compare poststapedectomy hearing gain in study-eligible patients who had received a Teflon (fluoroplastic ASTM F 754) prosthesis (study group; n = 76) with hearing gain achieved in a matched group (by age, sex, and degree of hypoacusis) of patients who had received a Schuknecht type wire/Teflon prosthesis (control group; n = 70). All procedures had been performed by the authors at our institution between Jan. 2, 1994, and Dec. 31, 1997. Airway averages at low, medium, and high frequencies were estimated on the basis ofpre- and postoperative audiologic evaluations, as were total air-bone gaps at 7 frequencies between 125 and 8, 000 Hz. We found that the study group achieved a significantly greater degree of hearing gain at 125 and 250 Hz and significantly better closure of the air-bone gap at 250, 500, 1,000, 2,000, and 4,000 Hz. The hearing outcomes among patients in the study group were excellent. PMID- 16381134 TI - Primary carcinosarcoma of the helix of the ear. AB - We report our histologic and immunohistochemical findings in a rare case of cutaneous carcinosarcoma involving the helix of the ear. The tumor exhibited cellular features of both basal cell and squamous cell carcinoma and a malignant mesenchymal component that was consistent with malignant fibrous histiocytoma. The epithelial component exhibited a positive immunohistochemical reaction to cytokeratin and a negative reaction to vimentin, whereas the mesenchymal component showed a positive immunohistochemical reaction to vimentin and a negative reaction to cytokeratin. To the best of our knowledge, this is only the third reported case of a carcinosarcoma of the ear and the second case in which it developed on the helix. PMID- 16381135 TI - Supernumerary nostril with congenital cataract. AB - Supernumerary nostril is a very rare congenital anomaly. It can be unilateral or bilateral, and it sometimes occurs in the presence of other congenital deformities. Behind the external opening of a supernumerary nostril is a small accessory nasal cavity, which may or may not communicate with the normal nasal cavity on the same side. We describe a new case in which the supernumerary nostril with a small accessory nasal cavity, which did not communicate with the normal nasal cavity on the same side, appeared in a young girl who also had microcornea and congenital cataract. The accessory nasal cavity was successfully removed surgically. We believe that this case may represent the first reported case of a supernumerary nostril with a congenital cataract on the same side. We also discuss the hypotheses that have been proposed to explain supernumerary nostrils. PMID- 16381136 TI - Extramedullary plasmacytoma arising from the nasal septum. AB - We report a rare case of extramedullary plasmacytoma of the nasal septum in a 65 year-old woman. She presented with a 2-month history of left-sided nasal obstruction and intermittent blood-tinged nasal crusting. Nasal endoscopy revealed that a dark-red mass had arisen from the nasal septum; no evidence of invasion to adjacent tissues was seen. A biopsy specimen was diagnosed as a plasmacytoma (kappa light chain-type). Serum and urine electrophoresis failed to detect any myeloma component or Bence Jones protein. All other screening tests to rule out multiple myeloma were negative. These findings confirmed the diagnosis of extramedullary plasmacytoma. The mass was completely removed via an endoscopic approach. No recurrence was noted at the 2-year follow-up. PMID- 16381137 TI - Surgical emphysema in the neck as a result of a dental procedure. AB - We report the development of subcutaneous emphysema in a middle-aged woman that occurred several hours after she had undergone a dental restoration procedure. The patient presented to the emergency department, and she was admitted for observation and prophylactic antibiotic coverage. She recovered in 3 days without further intervention and was discharged. PMID- 16381138 TI - Spontaneous retropharyngeal and cervical emphysema: a rare singer's injury. AB - Spontaneous retropharyngeal and cervical emphysema is rare. We describe a case that was unusual in its etiology: the result of singing. Although this condition is usually benign, hospital admission for close observation and supportive therapy is prudent. PMID- 16381139 TI - Myxolipoma of the epiglottis in an adult: a case report. AB - Lipomas of the larynx are rare. When they do occur, they are important because some require that the surgeon perform a tracheotomy to prevent a fatal airway obstruction. We report a case of myxolipoma-that is, a lipoma with foci of myxomatous degeneration-of the epiglottis in a 56-year-old woman. We successfully treated the tumor with intubation and surgery; tracheotomy was not necessary. To the best of our knowledge, myxolipoma of the epiglottis in an adult has not been previously reported in the English-language literature. PMID- 16381140 TI - Schwannoma of the larynx: a case report. AB - Neurogenic tumors of the larynx are rare. We report a case of supraglottic schwannoma in a 59-year-old woman. We excised the tumor via a lateral thyrotomy approach. PMID- 16381141 TI - Impaired sustained attention and error awareness in traumatic brain injury: implications for insight. AB - The processes of error awareness and sustained attention were investigated in 18 traumatic brain injury (TBI) individuals and 16 matched control participants. In Experiment 1, we found that: (1) in comparison to controls, TBI participants displayed reduced sustained attention and awareness of error during the Sustained Attention to Response Task; (2) degree of error awareness was strongly correlated with sustained attention capacity, even with severity of injury partialed out; and (3) that error feedback significantly reduced errors. We replicated the finding of a correlation between error awareness and sustained attention capacity in Experiment 2 with a separate sample of 19 TBI participants and 20 controls. We conclude that TBI leads to impaired sustained attention and error awareness. The finding of a significant relationship between these two deficits in TBI suggests there may be a link between these two processes. Feedback on error improves sustained attention performance of control and TBI participants. PMID- 16381142 TI - The contribution of the left and right hemispheres to early recovery from aphasia: a SPECT prospective study. AB - This prospective study examined the relationship between post-stroke recovery of aphasia and changes in cerebral blood flow (CBF). To address the question of right hemisphere (RH) involvement in restitution of language, we tested the hypothesis that the increase in perfusion of the RH is crucial for early recovery from aphasia. Twenty-four right-handed patients with acute aphasia following left hemisphere (LH) ischaemic stroke were examined twice with a six-month interval. At each session CBF and language scores were measured on the same stroke patients. Language was measured by selected tasks derived from the Boston Diagnostic Aphasia Examination (BDAE). The SPECT scans were obtained using (99m)Tc-ECD on a triple-head gamma camera Multispect-3. Although initial CBF measured for the whole group of aphasic patients was not a predictor for future language recovery for either hemisphere, increased perfusion of the RH during a six-month interval was found to parallel the recovery of aphasic disorders. There was a correlation between the change in the right parietal CBF (but not the left) and a change in numerous language abilities. Nevertheless, only CBF values on the left predicted performance on the language tests at initial and follow-up examinations. When the area damaged on structural imaging was excluded from perfusion analysis, only subcortical CBF change on the left showed a positive correlation with language improvement. Thus, the cerebral mechanism associated with early recovery from aphasia is a dynamic and complex process that may involve both hemispheres. Probably this mechanism involves functional reorganisation in the speech-dominant (damaged) hemisphere and regression of haemodynamic disturbances in the non-dominant (structurally intact) hemisphere. PMID- 16381143 TI - Effects of a neuropsychological rehabilitation programme on schizophrenic patients' subjective perception of improvement. AB - Cognitive and neuropsychological deficits have long been recognised as part of the schizophrenic syndrome. As these deficits persist after acute psychosis subsides and contribute to the social and occupational difficulties often experienced by individuals with schizophrenia, researchers have begun to consider the applicability of neuropsychological rehabilitation techniques to the treatment of schizophrenic neuropsychological deficits. Although the effectiveness of cognitive rehabilitation programmes in schizophrenia has been addressed, no attention has been afforded to whether the trained patient perceives improvement of the cognitive processes instructed. The present study utilised a treatment versus control group comparison design to evaluate the effect of a neuropsychological rehabilitation programme on the perception of improvement both in schizophrenic patients and their therapists. Twenty-four participants diagnosed with schizophrenia participated in the study. One half of the sample was trained by means of Attention Process Training (APT) and the rest of the patients comprised the control group. No significant differences in perception of improvement were found between treatment and control group, and trained patients found the APT more useful than did their therapists. PMID- 16381144 TI - Using errorless learning to treat letter-by-letter reading: contrasting word versus letter-based therapy. AB - Some pure alexic readers have been shown to activate lexical and semantic knowledge under brief presentation conditions. This ability is not seen when letter-by-letter reading accuracy is high or the reading impairment is very severe. It is also unlikely to occur under normal untimed presentation because the pure alexic will make deliberate use of their letter-by-letter strategy. This paper presents data from a moderately severe letter-by-letter reader, FD, who had visual processing problems affecting reading. He also had other mild aphasic characteristics. FD showed implicit reading abilities under brief presentation conditions, being able to make lexical decisions and semantic categorisations well above chance. FD was given two therapy programmes, the first, whole word therapy to exploit this implicit ability and the second to improve letter-by letter accuracy and speed. FD showed some improvement in reading ability after both therapy programmes, particularly for words of personal interest to him. His letter naming accuracy and reading of visually similar words were the most resistant to change. A striking effect of therapy was the cessation of FD's letter-by-letter reading and the emergence of some of the characteristics of deep dyslexia. Even when therapy concentrated on letter accuracy, FD did not revert back to his original letter-by-letter reading strategy. The results are discussed with reference to the two theories of pure alexia. Some conclusions are drawn about the need for therapists to examine and exploit all residual reading skills when devising therapeutic programmes. PMID- 16381145 TI - The effect of residential and agricultural runoff on the microbiology of a Hawaiian ahupua'a. AB - The objective of this project was to study the relationship between environmental runoff and the incidence of antibiotic-resistant microorganisms (ARMO) in freshwater streams. Five water systems along the windward coast of the island of O'ahu were evaluated. Samples were collected from sites upstream of residential or agricultural areas, throughout these areas, and at sites of entrance into oceans or bays. It was hypothesized that the incidence of ARMO would increase as the stream received runoff from residential and agricultural areas. The percentage of ARMO did not increase as the streams passed through residential or agricultural areas. Surprisingly, pristine sites, well upstream from residential or agricultural areas, contained bacteria resistant to at least one antibiotic. Areas most affected by runoff did not show a significant increase in the incidence of antibiotic-resistant organisms, suggesting that the incidence of antibiotic resistance is not simply a function of contamination with agricultural or residential runoff. The correlation of antibiotic resistance with heavy metal resistance was evaluated, because others (Fasim et al., 1999; Lazar et al., 2002; Nies, 1999) have shown that antibiotic and heavy metal resistance are each carried on extrachromosomal plasmids. The vast majority of ARMO were also resistant to concentrations of heavy metals reported in the sediments of indicator streams (Waihee, system III), suggesting that an antibiotic-resistant bacterium has a high probability of having dual resistance to a heavy metal. A 3.2-kb plasmid (pSTAMP) was isolated from a bacterium with dual antibiotic and heavy metal resistance. Further analysis of the plasmid is currently in progress. PMID- 16381146 TI - Antibiotic resistance from wastewater oxidation ponds. AB - In an extensive, multiyear study of antibiotic resistance from wastewater oxidation ponds, five mobile home park wastewater oxidation ponds in Clarke and Oconee counties were shown to be discharging high numbers of antibiotic-resistant bacteria into the waterways of North Georgia. This effluent contributed to higher nitrogen, phosphorus, and fecal coliform levels in creeks downstream from the ponds. A survey of residents revealed that many people did not complete their antibiotic prescriptions, and the majority flushed leftover antibiotic medications down the toilet. In the pond discharges, resistance was found to eighteen antibiotics: amikacin, amoxicillin/clavulanic acid, ampicillin, apramycin, cefoxitin, ceftiofur, ceftriaxone, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, imipenem, kanamycin, naladixic acid, streptomycin, sulphamethoxazole, trimethoprim/sulphamethoxazole, and tetracycline. The discharged bacteria contained both integrons and plasmids, the latter being transferable to a laboratory strain of Escherichia coil (E. coli). A turtle was found living at a pond discharge site with multiply-antibiotic-resistant bacteria in its feces. Last year, RNA fingerprinting conclusively documented the survival of three multiply-resistant important pathogenic bacteria. Ceftriaxone-resistant Stenotrophomonas maltophilia and Pseudomonas aerogenosa and a ciprofloxacin resistant E. coli were traced through oxidation pond stages and into the discharge, thus documenting that the pathogens survived the treatment process. In addition, a potential pathogen, a serotype group D Salmonella spp., was found in the discharge. In this study, tetracycline-resistance genes C and G were detected in the first and second stages of the oxidation pond and the discharge went directly into the environment. These genes are generally found in intestinal bacteria, so it can be inferred that they are from a human source. Antimicrobial residue from the beta-lactam family of antibiotics was found in all oxidation pond stages and in the creek above the pond. Tetracycline residue was found in the first and second stages of the pond. In addition to the antibiotics, genes coding for antibiotic resistance and the antibiotics themselves were documented to survive oxidation pond treatment. Tetracycline-resistant genes were identified in the oxidation pond stages and in the discharge going into the environment. A model was also developed to study oxidation pond function in the laboratory. A biofilm was created using a highly antibiotic-resistant Salmonella typhimurium 3/97, and pond water was added. The biofilm was processed via a rotating disk bioreactor specifically designed to study biofilms in nature, but with conditions that were more favorable to bacterial inhibition than those in nature. Cultures revealed that, under these optimal conditions, S. typhimurium 3/97 was still present in this in vitro system. Thus, the competitive inhibition process that helps to remove bacteria in oxidation ponds did not effectively remove an important bacterium, S. typhimurium 3/97, in this mock oxidation pond. The bioreactor model developed in this study can be used to further investigate discharges from oxidation ponds. From this data, it is apparent that the problem is two-fold. A cost-effective technique must be developed that inactivates antibiotic-resistant bacteria in oxidation pond discharges and also removes the antibiotics. A public awareness campaign was initiated by the author to encourage proper use and disposal of antibiotics, as flushing them is a common practice in the United States. PMID- 16381147 TI - Calcium carbonate phosphate binding ion exchange filtration and accelerated denitrification improve public health standards and combat eutrophication in aquatic ecosystems. AB - Cultural eutrophication, the process by which a lake becomes rich in dissolved nutrients as a result of point and nonpoint pollutant sources, is a major cause of the loss of natural lake ecosystems throughout the world. The process occurs naturally in all lakes, but phosphate-rich nutrient runoff from sources such as storm drains and agricultural runoff is a major cause of excess phosphate-induced eutrophication. Especially in Madrona Marsh, one of the last remaining vernal marshes in the greater Los Angeles area, California, cultural eutrophication has become a major problem. In this study, calcium carbonate was found to be an excellent phosphate binder, reducing up to 70% of the phosphates in a given sample of water, and it posed relatively negligent ecological repercussions. This study involved the testing of this principle in both the laboratory and the real ecosystem. A calcium carbonate lacing procedure was first carried out to determine its efficacy in Madrona Marsh. Through this, ammonia was found to interfere with the solubility of calcium carbonate and therefore to be a hindrance to the reduction of phosphate. Therefore, various approaches for reduction of ammonia were tested, including aeration, use of fiber growth media, and plants, mainly Caulerpa verticellata, chosen for it hardiness, primarily in an attempt to increase population of Nitrobacter and Nitrosomonas. All were successful in moderately reducing ammonia levels. In addition, soil sampling, sediment analysis, microscopic plant analysis, microorganism and macroinvertebrate identification, and rate law formulations were conducted. The effect of phosphate and ammonia reduction on the populations of enterobacteria was also an important focus of this experiment. Varying concentrations of phosphate, ammonia, and calcium carbonate in conjunction with phosphate were tested in Madrona Marsh to determine their effects on the populations of enteropathogens on nonspecific blood agar, MacConkey agar, and Hektoen agar. Initial analyses suggest a strong correlation between phosphate concentrations and bacterial populations; a 66% decrease in phosphate resulted in a 35% reduction in bacterial populations and a 45% reduction in enteropathogenic populations. Likewise, a strong correlation was shown between calcium carbonate concentrations and bacterial reduction greater than that which can be attributed to the phosphate reduction alone. This was followed by the construction of various phosphate binding calcium carbonate filters, which used the ion exchange principle, including a spring loading filter, PVC pipe filter, and a galvanized filter. All were tested with the aid of Stoke's law formulation. The experiment was extremely successful in designing a working phosphate-binding and ammonia reducing filter, and a large-scale agitator-clarifier filter system is currently being planned for construction in Madrona Marsh; this filter will reduce phosphate and ammonia levels substantially in the following years, bringing ecological, economical, and health-related improvements to the overall ecosystem and habitat. PMID- 16381148 TI - Bottled water: how safe is it? AB - Sales of bottled water have increased dramatically in recent years, with worldwide sales of more than dollars 35 billion, largely because of the public perception of purity and safety and public concern about the quality of tap water. Presently, there are no Food and Drug Administration (Washington, D.C.) recommendations regarding temperature and duration of storage for bottled water once it is opened and used. The objectives of this study were to examine the effects of time and storage temperature on bacterial growth and characterize the types of microorganisms contaminating bottled water after drinking once from the bottle. Bottled and tap water were tested using standard microbiology culture techniques. The bacterial count in bottled water increased dramatically, from less than 1 colony per milliliter (col/mL) to 38,000 col/mL over 48 hours of storage at 37 degrees C. Bacterial growth was markedly reduced at cold temperatures (refrigeration) compared with room temperature, with 50% fewer bacterial colonies in 24 hours and 84% fewer colonies in 48 hours. Interestingly, tap water resulted in only minimal growth, especially at cold temperatures (< 100 col/mL at 48 hours). These findings may be useful to increase public awareness and development of guidelines on storage temperature and expiration time for bottled water once it is opened and used. PMID- 16381149 TI - Laboratory evaluation of thermophilic-anaerobic digestion to produce Class A biosolids. 1. Stabilization performance of a continuous-flow reactor at low residence time. AB - There is increasing interest in the United States in producing biosolids from municipal wastewater treatment that meet the criteria for Class A designation established by the U.S. Environmental Protection Agency. Class A biosolids are intended to be free of pathogens and also must meet requirements for reduction of the vector-attraction potential associated with untreated sludge. High temperature processes are considered to produce Class A biosolids if the combination of operating temperature and treatment time exceeds minimum criteria, but this option is not applicable to mixed, continuous-flow reactors. Such reactors, or any combination of reactors that does not meet the holding time requirement at a specific temperature, must be demonstrated to inactivate pathogens to levels consistent with the Class A criteria. This study was designed to evaluate pathogen inactivation by thermophilic anaerobic digestion in a mixed, continuous-flow reactor followed by batch or plug-flow treatment. In this first of a two-part series, we describe the performance of a continuous-flow laboratory reactor with respect to physical and chemical operating parameters; microbial inactivation in the combined continuous-flow and batch treatment system is described in the second part. Sludges from three different sources were treated at 53 degrees C, while sludge from one of the sources was also treated at 55 and 51 degrees C. Relatively short hydraulic retention times (four to six days) were used to represent a conservative operating condition with respect to pathogen inactivation. Treatment of a fermented primary sludge led to an average volatile solids (VS) destruction efficiency of 45%, while VS destruction for the other two sources was near or below 38%, the Class A criterion for vector attraction reduction. Consistent with other studies on thermophilic anaerobic digestion of sludges at short residence times, effluent concentrations of volatile fatty acids (VFAs) were relatively high. Also consistent with other studies, the most abundant VFA in the effluent was propionate. Gas production ranged from 0.3 to 0.5 m3/kg VS fed and from 0.8 to 1.3 m3/kg VS destroyed. PMID- 16381150 TI - Laboratory evaluation of thermophilic-anaerobic digestion to produce Class A biosolids. 2. Inactivation of pathogens and indicator organisms in a continuous flow reactor followed by batch treatment. AB - Thermophilic-anaerobic digestion in a single-stage, mixed, continuous-flow reactor is not approved in the United States as a process capable of producing Class A biosolids for land application. This study was designed to evaluate the inactivation of pathogens and indicator organisms in such a reactor followed by batch treatment in a smaller reactor. The combined process was evaluated at 53 degrees C with sludges from three different sources and at 51 and 55 degrees C with sludge from one of the sources. Feed sludge to the continuous-flow reactor was spiked with the pathogen surrogates Ascaris suum and vaccine-strain poliovirus. Feed and effluent were analyzed for these organisms and for indigenous Salmonella spp., fecal coliforms, Clostridium perfringens spores, and somatic and male-specific coliphages. No viable Ascaris eggs were observed in the effluent from the continuous reactor at 53 or 55 degrees C, with greater than 2 log removals across the digester in all cases. Approximately 2-log removal was observed at 51 degrees C, but all samples of effluent biosolids contained at least one viable Ascaris egg at 51 degrees C. No viable poliovirus was found in the digester effluent at any of the operating conditions, and viable Salmonella spp. were measured in the digester effluent in only one sample throughout the study. The ability of the continuous reactor to remove fecal coliforms to below the Class A monitoring limit depended on the concentration in the feed sludge. There was no significant removal of Clostridium perfringens across the continuous reactor under any condition, and there also was limited removal of somatic coliphages. The removal of male-specific coliphages across the continuous reactor appeared to be related to temperature. Overall, at least one of the Class A pathogen criteria or the fecal coliform limit was exceeded in at least one sample in the continuous-reactor effluent at each temperature. Over the range of temperatures evaluated, the maximum time required to meet the Class A criteria by batch treatment of the continuous-reactor effluent was 1 hour for Ascaris suum and Salmonella spp. and 2 hours for fecal coliforms. PMID- 16381151 TI - Effect of high influent sulfate on anaerobic wastewater treatment. AB - A laboratory-scale study was conducted using a completely mixed reactor with a constant influent-total-organic carbon (TOC) of 3750 mg/L to evaluate the effect of increasing influent-sulfate levels on anaerobic-treatment performance. The sulfate levels were increased stepwise from 333 to 666, 1000, 1333 and 1666 mg S/L. The results showed that an elevation of influent sulfate actually increased the TOC removal efficiency as long as the produced sulfide level did not induce toxicity. At 1333 mg S/L influent sulfate, the produced dissolved sulfide was 613 mg S/L (free sulfide = 228 mg S/L), which started to impose toxicity to the methane-producing bacteria (MPB). It was also found that the percent electron flow to the sulfate-reducing pathway increased with the increasing influent sulfate, but the direction toward the methanogenesis was correspondingly reduced. Nevertheless, under the experimental conditions tested, the majority of the influent organics was still degraded through the methanogenic pathway. Through this study, an oxidation-reduction-potential (ORP)-based oxygenation process was developed for online oxidation of sulfide in recirculating biogas. With controlled oxygen injection to raise the reactor's ORP by 25 mV, the residual sulfide in the reactor was almost totally eliminated. In case of over oxygenation, any excess oxygen was quickly consumed by the facultative organisms in the reactor, thereby imposing no toxicity to the MPB. PMID- 16381152 TI - Simulation evaluation of the effects of non-uniform flow and degradation parameter uncertainty on subsurface-flow constructed wetland performance. AB - Although constructed wetland treatment systems have been used in a variety of applications, uncertainty in adequately determining flow conditions or hydraulic residence times ("hydraulic efficiencies") and degradation model parameters remains a problem with their design. Breakthrough or impulse-type tracer studies in constructed wetlands often result in residence-time distributions exhibiting long skewed "tails" suggesting multiple flow channels or perhaps unrealistically large dispersion factors. A fractional-flow analysis is developed here to quantify possible flow non-uniformity in a subsurface-flow constructed wetland and is then used to assess the effects of non-uniformity and degradation model parameter variability on constituent (for example, chemical oxygen demand) removal. A model application to tracer data developed previously demonstrates how flow non-uniformity alone can account for significant "tailing" and can be related to even moderate estimated dispersion numbers. From the analysis, it is evident that flow non-uniformity is of greater concern than decay parameter uncertainty, and that, from a constructed wetland design and operation perspective, every effort should be made to ensure relative flow uniformity across the constructed wetland. PMID- 16381153 TI - Ion exchange treatment of rinse water generated in the galvanizing process. AB - A study was conducted of the viability of using the cationic exchange resins Amberlite IR-120 and Lewatit SP-112 to treat rinse water generated in the galvanizing process as well as acidic wastewater containing zinc (Zn) and iron (Fe). Solutions containing either 100 mg/L of Zn at pH 5.6 (rinse water) or Fe and Zn at concentrations of 320 and 200 mg/L at pH 1.5 (acidic water), respectively, were percolated through packed beds until the resins were exhausted. Breakthrough capacities obtained ranged between 1.1 and 1.5 meq metal/mL resin. The elution of metal and the regeneration of resins were performed with hydrochloric acid. The influence of the flowrate used during the loading stage was also studied, with 0.5 bed volumes/min (3.2 cm/min) found to be the optimum flowrate. PMID- 16381154 TI - An equilibrium model for chloride removal from recycled cooling water using the ultra-high lime with aluminum process. AB - Removal of chloride from recycled cooling water is needed to reduce corrosion and prolong equipment life. Laboratory experiments have demonstrated that the ultra high lime with aluminum (UHLA) process has the ability to achieve high chloride removal efficiency from recycled cooling water. In an effort to further understand the behavior of chloride in the UHLA process, a fundamental model of the chemical processes was developed. The purpose of this paper is to describe this equilibrium model and present values for solubility products of precipitated solids that have not been investigated previously. The model was based on PHREEQC and a new program called INVRS K was integrated with PHREEQC to calculate values of unknown or poorly defined equilibrium or kinetic constants using a Gauss Newton nonlinear regression routine. Model predictions indicated that the results could be best described by assuming the formation of a solid solution of calcium chloroaluminate (Ca4Al2Cl2OH12), tricalcium hydroxyaluminate (Ca3Al2OH12), and tetracalcium hydroxyaluminate (Ca4Al2OH14). PMID- 16381155 TI - Development of a new model for batch sedimentation and application to secondary settling tanks design. AB - Assuming that settling takes place in two zones (a constant rate zone and a variable rate zone), a model using four parameters accounting for the nature of the water-suspension system has been proposed for describing batch sedimentation processes. The sludge volume index (SVI) has been expressed in terms of these parameters. Some disadvantages of the SVI application as a design parameter have been pointed out, and it has been shown that a relationship between zone settling velocity and sludge concentration is more consistent for describing the settling behavior and for design of settling tanks. The permissible overflow rate has been related to the technological parameters of secondary settling tank by simple working equations. The graphical representations of these equations could be used to optimize the design and operation of secondary settling tanks. PMID- 16381156 TI - Presence of Aeromonas spp in water from drinking-water- and wastewater-treatment plants in Mexico City. AB - The frequency of Aeromonas spp in three wastewater-treatment plants (WWTPs) and two drinking-water plants (DWPs) in Mexico City was determined. Samples were taken throughout a year by the Moore's swab technique. A total of 144 samples were obtained from WWTPs and 96 from DWPs of both incoming and outflowing water. Aeromonas spp was isolated in 31% of the samples, from both kinds of sources. The technique used for the isolation of the pathogen was suitable for samples with high associate microbiota content and for those with a scarce microbial content. The presence of mesophilic-aerobic, coliform, and fecal-coliform organisms was investigated to determine whether there was any correlation with the presence of Aeromonas spp. Most samples from WWTP, which did not comply with the Mexican standards, had the pathogen, and some of the samples from the outflow of the DWP, which were within the limits set by the Mexican standards, also had Aeromonas spp. Most samples containing Aeromonas spp. had concentrations below 0.1 ppm residual chlorine, and the strains were resistant to 0.3 ppm, which supports the recommendation to increase the residual chlorine concentration to 0.5 to 1.0 ppm, as recommended by the Mexican standards. PMID- 16381157 TI - Modeling depth filtration of activated sludge effluent using a compressible medium filter. AB - A new filter, using a compressible-filter medium, has been evaluated for the filtration of secondary effluent. The ability to adjust the properties of the filter medium by altering the degree of the medium compression is a significant departure from conventional depth-filtration technology. Unlike conventional filters, it is possible to optimize the performance of the compressible-medium filter (CMF) by adjusting the medium properties (i.e., collector size, porosity, and depth) to respond to the variations in influent quality. Because existing filter models cannot be used to predict the performance of the CMF, a new predictive model has been developed to describe the filtration performance of the CMF and the effect of medium-compression ratio. The model accounts for the fact that the properties of the filter medium change with time and depth. The model, developed for heterodisperse suspensions and variable influent total suspended solids concentrations, can be used to predict all possible phases of filtration (i.e., ripening, constant removal, and breakthrough). A hyperbolic-type, second order, nonlinear, partial-differential equation was derived to model the CMF. The equation was solved using the finite-difference numerical method. The accuracy of the numerical method was tested by a sensitivity analysis and a convergence test. The model is first-order accurate with respect to medium depth and time. Field data were obtained for the filtration of settled secondary effluent using a CMF with a capacity of 1200 m3/d. Model predictions were compared with observed performance from filter runs conducted at medium-compression ratios between 15 and 40% and filtration rates from 410 to 820 L/m2 min. The difference between the observed and the predicted values was found to be within 0 to 15%. PMID- 16381158 TI - Protocol for early detection and evaluation of inhibitory wastewater using combined aerobic respirometric and anaerobic batch techniques. AB - Faced with the task of treating significant volumes of complex industrial wastewaters, the biological components of municipal wastewater treatment plants are operating under the risk of toxic or inhibitory contaminants from the industrial effluents that may be detrimental to their operation. This might lead to undesirable effluent toxicity and/or result in permit violations. Therefore, there is a need for upset early warning systems that can protect full-scale plants from toxic or inhibitory constituents in the incoming wastewaters. This study focused on the development of a protocol for rapid detection of potentially toxic inhibitory or toxic wastewaters using combined aerobic respirometric and anaerobic batch techniques. Aerobic respirometers equipped with automated data acquisition systems were used as potential early warning devices. The inhibition effect on carbon and nitrogen oxidation was assessed. The degree of inhibition was evaluated as the concentration causing 50% reduction in microbial activity, which was estimated by an inhibition model. Anaerobic toxicity assays were also conducted to evaluate the inhibitory effects of the toxic compounds to anaerobic inocula obtained from a master culture reactor fed with ethanol. The developed protocol for early detection of toxicity was validated using wastewater samples from a biotechnology industry and a food processing industry, and pure chemicals such as furfural and phenol. Varying degrees of sensitivity were observed in the study when different groups of microorganisms, wastewater samples, and chemicals were tested. The comparison of aerobic and anaerobic inhibition suggested the importance of using both aerobic and anaerobic cultures to maximize the necessary sensitivity of the protocol. PMID- 16381159 TI - Evidence-based decision making in agricultural health and safety. PMID- 16381160 TI - Using fault tree analysis to identify contributing factors to engulfment in flowing grain in on-farm grain bins. AB - Findings reported by researchers at Illinois State University and Purdue University indicated that since 1980, an average of eight individuals per year have become engulfed and died in farm grain bins in the U.S. and Canada and that all these deaths are significant because they are believed to be preventable. During a recent effort to develop intervention strategies and recommendations for an ASAE farm grain bin safety standard, fault tree analysis (FTA) was utilized to identify contributing factors to engulfments in grain stored in on-farm grain bins. FTA diagrams provided a spatial perspective of the circumstances that occurred prior to engulfment incidents, a perspective never before presented in other hazard analyses. The FTA also demonstrated relationships and interrelationships of the contributing factors. FTA is a useful tool that should be applied more often in agricultural incident investigations to assist in the more complete understanding of the problem studied. PMID- 16381161 TI - Barriers to research participation identified by rural people. AB - It has been suggested anecdotally that rural people are less receptive to participating in research than other populations. Proposed reasons include culture, knowledge, attitudes, and barriers. Barriers to health care may also be barriers to research participation. A random sample of 5,000 households from a sampling frame of 45,000 property owners in a rural upstate New York county was selected. This article is a report of development of a barrier scale from the findings of 865 completed surveys. The survey in this study contained 100 questions and was adapted from a pre-existing survey of public attitudes regarding willingness to participate in medical research. Factor analysis was utilized to isolate a "barriers to participation in research" scale. Comparison of demographics and perceived barriers to participation were completed. Those who were classified as younger than the median sample age and male scored significantly lower on the barrier scale, indicating more barriers to participation in health care research. Those with the highest perception of barriers were among the least willing to participate as research subjects. The findings inform assumptions that researchers make about barriers to research, and strategies are suggested to remove such barriers. Opening the doors to inclusion of rural people in health research studies will ultimately result in improved individual and community health in rural places. PMID- 16381162 TI - Longitudinal analysis of pulmonary dysfunction in the initial years of employment in the grain industry. AB - The negative health effects of exposure to grain dust have previously been examined, but few studies have observed the effects on newly hired employees. Young grain workers are of interest because changes in pulmonary function may occur after a short duration of employment, and because older grain workers may represent a survivor population. The New Grain Workers Study (NGWS), a longitudinal study of 299 newly hired male grain industry workers, was conducted between 1980 and 1985. The objectives were to determine the effects of employment in the grain industry on pulmonary function. Pre-employment physical examinations and pulmonary function tests were conducted on subjects at the Division of Respiratory Medicine, Department of Medicine, Royal University Hospital, University of Saskatchewan. The Grain Dust Medical Surveillance Program (GDMSP) was a Labour Canada program that began in 1978. All subjects were grain workers employed in the grain industry in Saskatchewan. All subjects completed a respiratory symptoms questionnaire and underwent pulmonary function testing. Baseline observations were recorded every three years between 1978 and 1993. Data were available on 2184 grain workers. Generalized estimating equations were used to fit marginal and transitional multivariable regression models to determine the effects of grain dust exposure on pulmonary function. Marginal and transitional models were then compared. Height, exposure weeks, and previous FVC were predictive of FVC in the NGWS, while exposure weeks and previous FEV1 were predictive of FEV1. These models, as well as a transitional regression model built using the GDMSP data, were used to compute predicted mean annual decline inpulmonary function. Non-smoking grain workers in the NGWS had the highest pulmonary function test values, but also had the greatest predicted annual decline in pulmonary function. Ever-smoking grain workers in the GDMSP had the lowest pulmonary function test values. Non-smoking grain workers in the GDMSP had the least predicted annual decline in pulmonary function. PMID- 16381163 TI - Intellectual property rights, agriculture, and rural communities. AB - The global extension of intellectual property rights (IPRs) in agriculture and the simultaneous move towards evolving community and farmers' rights have significant implications for the future of rural communities. While IPRs are based on individual interests, the movement for granting farmers' rights is becoming diluted towards assigning ownership rights in a similar fashion as IPRs. It is important to reframe the debate to ensure that communities can claim their role in national and international negotiations. PMID- 16381164 TI - Sowing the seeds of hope: providing regional behavioral health supports to the agricultural population. AB - In 1999, project leaders from seven states (i.e., Iowa, Kansas, Minnesota, Nebraska, North Dakota, South Dakota, and Wisconsin) began to share ideas and resources for providing behavioral health assistance (i.e., mental health counseling and addictions services) to stressed farmers, ranchers, farm workers, and their families. The seven states are among those most impacted by the farm crisis of the 1980s and again by low commodity prices and disasters such as droughts and floods in the 1990s. Project leaders conferred in monthly telephone conference calls and by 2001 began meeting in semi-annual face-to-face meetings to formally agree on a mission, program components, and management structure. Administrative functions were transferred from the Wisconsin Office of Rural Health and Wisconsin Primary Healthcare Association, to AgriWellness, Inc., a regional nonprofit corporation founded to provide technical assistance, grant writing, training of service providers, and other administrative supports. The Sowing the Seeds of Hope program has become a model for the provision of behavioral health supports for the agricultural population, including development of farm stress telephone hotlines, provision of confidential and affordable outpatient mental health and substance abuse counseling, training of professional providers in agricultural behavioral health, training of indigenous farm and rural residents as outreach workers who can respond to disasters of all types, and weekend educational retreats for farm residents. The program has achieved economy of scale by sharing expertise across state boundaries and the formation of a regional administrative structure. Yet, many challenges exist, the greatest of which is obtaining ongoing permanent support for the increasing numbers of uninsured and underinsured people involved in agriculture. PMID- 16381165 TI - A study of the impact and efficacy of health fairs for farmers. AB - The purpose of this study was to evaluate the effectiveness of two regional multi disciplinary farm health fairs, pairing cardiovascular and agricultural health and safety risk factor assessments and education, in agricultural regions in southern Minnesota in 1999 and 2001. This study explores the rationales and motives cited as having an influence on: (1) work practice and lifestyle adaptations, (2) resistance to such changes, and (3) compliance with health fair recommendations. Evaluation took the form of standardized telephone surveys and callbacks with an open-ended component administered by a parish nurse interviewer and conducted six months following each of the fairs. Participants had received individualized work practice and lifestyle recommendations based upon their medical results and questionnaire responses. Of the 378 total attendees at the two farm health fairs, 272 (72%) participated by filling out on-site questionnaires, and 284 (75%) completed a post-fair interview. Participants who were actively farming totaled 237 (63%) of all participants. A majority of those interviewed on callback (78%) reported either work safety or lifestyle changes, while 47% of those actively farming claimed varying work safety changes as a result of the fair. The conclusions drawn from the follow-up interviews include: (1) the two farm health fairs resulted in a substantial number of participants positively modifying lifestyle and work practices, (2) the stand-alone farm health fairs were effective in attracting farmers, and (3) low-level stressors identified in the participants could be effectively addressed by knowledgeable rural health care resources such as parish nursing. PMID- 16381166 TI - [M. Nencki Institute of Experimental Biology]. PMID- 16381167 TI - [Letter to Minister of Science and Information Technology]. PMID- 16381168 TI - [Epigenome and cancer: new possibilities of cancer prevention and therapy?]. AB - In recent years it became evident that epigenetic control of gene expression plays important role in the development of cancer. Epigenetic alterations occur throughout all stages of carcinogenesis, what makes them excellent targets for both chemoprevention and, at later stages, chemotherapy. Two changes integral to epigenetic transcriptional control are DNA methylation and covalent modifications of histone proteins. In cancer cells, genome-wide hypomethylation is accompanied by local DNA hypermethylation with unique patterns exhibited by each tumor type. Hypermethylation of promoter regions of classic tumor suppressor genes in human cancers is at least as common as mutation as a mechanism for their inactivation. The second major layer of epigenetic transcriptional control is modification of histone proteins, particularly their acetylation. These two ways of epigenetic control are integrally linked. In this review we summarize the current knowledge about epigenetic events related to cancer and discuss the possibility of using them as chemotherapy or chemoprevention targets. PMID- 16381169 TI - [Tyrosine kinases. New target of anticancer therapy]. AB - Recently, clinical studies of new drugs development to target specific forms of cancer were reported. Herceptin, a monoclonal antibody against the Her2/neu receptor tyrosine kinase, prolonged the survival of women with Her2/neu positive metastatic breast cancer. STI571, a small molecule inhibitor of the BCR/ABL, c Kit and platelet derived growth factor receptor tyrosine kinase, produced pronounced clinical responses in patients with BCR/ABL positive chronic myeloid leukemia and c-Kit positive gastrointestial stromal tumors. In order to consider the use of the inhibitor of tyrosine kinases activity as anticancer drug, their mechanisms of the oncogenic activation and their impact on tumor transformation should be studied. The treatment with tyrosine kinase inhibitors such as STI571 or herceptin was a spectacular clinical success which stimulated research on the structure and function of both kinases and their inhibitors. PMID- 16381170 TI - [HMGA proteins and their genes as a potential neoplastic biomarkers]. AB - HMGA proteins and their genes are described in this article. HMGA proteins reveal ability to bind DNA in AT-rich regions, which are characteristic for gene promoter sequences. This interaction lead to gene silencing or their overexpression. In normal tissue HMGA proteins level is low or even undetectable. During embriogenesis their level is increasing. High HMGA proteins level is characteristic for tumor phenotype of spontaneous and experimental malignant neoplasms. High HMGA proteins expression correlate with bad prognostic factors and with metastases formation. HMGA genes expression can be used as a marker of tumor progression. Present studies connected with tumor gene therapy based on HMGA proteins sythesis inhibition by the use of viral vectors containing gene encoding these proteins in antisence orientation, as well as a new potential anticancer drugs acting as crosslinkers between DNA and HMGA proteins suggest their usefulness as a targets in cancer therapy. PMID- 16381171 TI - [Influence of human gastrointestinal tract bacterial pathogens on host cell apoptosis]. AB - Several pathogenic bacteria are able to trigger apoptosis in the host cell, but the mechanisms by which it occurs differ, and the resulting pathology can take different courses. Induction and/or blockage of programmed cell death upon infection is a result of complex interaction of bacterial proteins with cellular proteins involved in signal transduction and apoptosis. In this review we focus on pro/anti-apoptotic activities exhibited by two enteric pathogens Salmonella enterica, Yersinia spp. and gastric pathogen Helicobacter pylori. We present current knowledge on how interaction between mammalian and bacterial cell relates to the molecular pathways of apoptosis, and what is the role of apoptosis in pathogenesis. PMID- 16381172 TI - [The proinflammatory activity of lipopolysaccharide]. AB - Lipopolysaccharide (LPS) is the major component of the outer membranes of the walls of gramnegative bacteria that protects bacterial cells against antibacterial agents. LPS is the heteropolimer consisting of two parts: the hydrophobic lipid A and hydrophilic polisaccharide. After lysis of bacterial cells LPS is released into the circulation and it is able to activate the immunological system by stimulation of monocytes, macrophages, neutrophils, blood platelets and endothelial cells. LPS can bind to plasma proteins and form complexes that increase toxic activity of LPS and affinity of LPS to cell receptors. The activation of immune cells by LPS leads to release of inflammatory mediators: cytokines, chemokines, enzymes, eicosanoids, adhesion agents and free radicals that are responsible for progression of inflammatory reactions and may induce pathophysiological processes including septic shock. PMID- 16381173 TI - [Thyroid transcription factors involved in the thyroid hormones biosynthesis]. AB - The thyroid hormone biosynthesis is dependent on the level of the expression and the correct action of the basic proteins involved in this process: TSH receptor (TSH-R), sodium-iodine symporter (NIS), thyroglobulin (Tg), thyroid peroxidase (TPO) and iodothyronine deodinases type 1 (D1) and type 2 (D2). Transcription of the genes coding these proteins is regulated by TSH via cAMP cascade. Despite the thyroid-specific transcription factors are necessary for the transcription regulation of TSH-R, NIS, TG, TPO, DIO1 and DIO2 genes. Three thyroid transcription factors Titf1/Nkx2-1, Foxel and Pax8 seem to be essential for thyroid development, growth and function. Disturbances of the expression of these transcription factors and their co-operation could be responsible for the congenital thyroid gland disorders as well as in thyroid cancerogenesis. PMID- 16381174 TI - [Diseases associated with protein aggregation]. AB - The so-called conformational diseases constitute a specific subtype of protein folding diseases that is characterized by abnormal aggregation of improperly folded polypeptides. This review describes a series of examples of such disorders and summarizes the present knowledge on their molecular pathophysiology and new therapeutic strategies. PMID- 16381175 TI - [Adaptor proteins of blood platelets]. AB - Adaptor proteins play a pivotal role in the regulation of signal transduction events elicited after the engagement of cell surface receptors. Platelets exhibit a number of integral membrane receptors capable of initiating a cellular response. These include collagen receptors, von Willebrand factor receptors, the fibrinogen receptor, and a number of G-protein coupled receptors, such as those for thrombin and ADP. The primary function of platelet receptors is the translation of externally applied signals into appropriate responses leading to platelet activation being a prerequisite for normal hemostasis. Multitude of signalling pathways described in platelets is based on the interaction of compounds of many different categories, such as transmembrane receptors, protein kinases, protein phoshatases, G-proteins, transmembrane and cytosolic adaptor proteins, phosphoinositides, cyclic AMP or GMP. Adaptor proteins lack intrinsic effector function, but contain distinct molecular domains, which mediate protein protein and protein-lipid interactions. These molecules thus serve as a scaffolding, around which effectors and their substrates are assembled into three dimensional signaling complexes. Adaptor proteins integrate receptor-mediated signals at intracellular levels and couple signaling receptors to cytosolic signaling pathways. While the function of adaptor proteins is well established in immune cells, the knowledge about their role in platelet activation is still at the onset Over the last decade numerous adaptor proteins have been identified in platelets and shown to be involved in accurate assembly of intracellular signaling complexes. Collagen-induced platelet intracellular signaling through GPVI resembles the functional response of B- and T-cell antigen receptors and is the best described in the literature. This review focuses on the structure and functional role of the most extensively studied adaptor proteins during platelet activation induced by physiological agonists. PMID- 16381176 TI - [Cystatins, thyropins and inhibitors homologous to propeptides of cysteine proteases]. AB - Protein inhibitors of proteolytic enzymes play an important role in regulating the activity of endogenous proteases and in host defense mechanisms against pathogens preventing the deleterious effects of exogenous proteases. In recent years a great interest in protein inhibitors of cysteine proteases has increased due to the extensive growth of knowledge about the contribution of cysteine proteases to pathological processes associated with many human diseases, as well as due to prospects for treatment of these disorders which may arise from the thorough understanding of their inhibitory mechanisms. This paper reviews the most important aspects of three families of cysteine protease inhibitors: cystatins, thyropins and inhibitors homologous to propeptides of cysteine proteases. Special attention is given to structural bases of the interactions between the inhibitors and their target enzymes. The paper presents a general characterization of the families according to the MEROPS classification of protease inhibitors, pointing out new members. PMID- 16381177 TI - [Non-homologous DNA end joining]. AB - DNA double strand breaks (DSB) are the most serious form of DNA damage. Repair of DSBs is important to prevent chromosomal fragmentation, translocations and deletions. Non-homologous end joining (NHEJ) is one of three major pathways for the repair of DSBs in human cells. In this process two DNA ends are joined directly, usually with no sequence homology, although in the case of same polarity of the single stranded overhangs in DSBs, regions of microhomology are utilized. NHEJ is typically imprecise, a characteristic that is useful for immune diversification in lymphocytes in V(D)J recombination. The main components of the NHEJ system in eukaryotes are the catalytic subunit of DNA protein kinase (DNA PKcs), Ku proteins, XRCC4, DNA ligase IV, and Artemis. This review focuses on the mechanisms an dregulation of DSB repair by NHEJ in mammalian cells. PMID- 16381178 TI - [Specific arginine mediated RNA recognition]. AB - In many biological systems substantial roles are played by interactions between amino acids and RNA. Among amino acids L-arginine seems to be particularly relevant, because the guanidinium group of arginine side chain can potentially form five hydrogen bonds with appropriately positioned acceptor groups of RNA. Extensive studies reveal that specific arginine recognition is achieved by many different RNAs over a broad range of binding affinities. Arginine is frequently found among amino acids in the nucleic acid-binding motifs in various proteins. For example, specific binding of the HIV-1 Tat protein to its RNA site (TAR) is mediated by a single arginine residue. Free arginine can be also bound by the guanosine site in the group I Tetrahymena ribosomal RNA intron catalytic centre, as well as by numerous RNA motifs, called arginine aptamers, which have been selected in vitro. PMID- 16381179 TI - [Application of molecular biology for the discovery of biosynthetic genes of polyketide and peptide antibiotics produced by actinomycetes]. AB - Actinomycetes are currently the main source of antibiotics. Genome sequencing reveals the presence in these organisms of multiple gene clusters for the synthesis of yet unidentified secondary metabolites. Technological advances in DNA isolation, cloning and sequencing, as well as development of bioinformatics, facilitate large scale search for new gene clusters in organisms with unknown genome sequence and in environmental DNA. Methods used for detection of polyketide synthase (PKS) and non-ribosomal peptide synthetase (NRPS) genes are described in this article. New PKS and NRPS genes give access to new biologically active natural products which can become drugs or substrates for chemical modifications. Even more inspiring is their use in combinatorial biosynthesis to produce a variety of compounds with rationally designed structures. PMID- 16381180 TI - Deep brain stimulation in movement disorders: stereotactic coregistration of two dimensional electrical field modeling and magnetic resonance imaging. AB - OBJECT: Adjusting electrical parameters used in deep brain stimulation (DBS) for dystonia remains time consuming and is currently based on clinical observation alone. The goal of this study was to visualize electrical parameters around the electrode, to correlate these parameters with the anatomy of the globus pallidus internus (GPI), and to study the relationship between the volume of stimulated tissue and the electrical parameter settings. METHODS: The authors developed a computer-assisted methodological model for visualizing electrical parameters (the isopotential and the isoelectric field magnitude), with reference to the stereotactic target, for different stimulation settings (monopolar and bipolar) applied during DBS. Electrical field values were correlated with the anatomy of the GPI, which was determined by performing stereotactic magnetic resonance imaging in one reference patient. By using this method it is possible to compare potential and electrical field distributions for different stimulation modes. In monopolar and bipolar stimulation, the shape and distribution of the potential and electrical field are different and depend on the stimulation voltage. Distributions visualized for patient-specific parameters can be subsequently correlated with anatomical information. The application of this method to one patient demonstrated that the 0.2-V/ mm isofield line fits best with the lateral GPI borders at the level of the stimulated contacts. CONCLUSIONS: The electrical field is a crucial parameter because it is assumed to be responsible for triggering action potentials. Electrical field visualization allows the calculation of the stimulated volume for a given isoline. Its application to an entire series of patients may help determine a threshold for obtaining a therapeutic effect, which is currently unknown, and consequently may aid in optimizing parameter settings in individual patients. PMID- 16381181 TI - Deep brain stimulation in Parkinson disease: a metaanalysis of patient outcomes. AB - OBJECT: Deep brain stimulation (DBS) to treat advanced Parkinson disease (PD) has been focused on one of two anatomical targets: the subthalamic nucleus (STN) and the globus pallidus internus (GPI). Authors of more than 65 articles have reported on bilateral DBS outcomes. With one exception, these studies involved pre- and postintervention comparisons of a single target. Despite the paucity of data directly comparing STN and GPI DBS, many clinicians already consider the STN to be the preferred target site. In this study the authors conducted a metaanalysis of the existing literature on patient outcomes following DBS of the STN and the GPI. METHODS: This metaanalysis includes 31 STN and 14 GPI studies. Motor function improved significantly following stimulation (54% in patients whose STN was targeted and 40% in those whose GPI was stimulated), with effect sizes (ESs) of 2.59 and 2.04, respectively. After controlling for participant and study characteristics, patients who had undergone either STN or GPI DBS experienced comparable improved motor function following surgery (p = 0.094). The performance of activities of daily living improved significantly in patients with either target (40%). Medication requirements were significantly reduced following stimulation of the STN (ES = 1.51) but did not change when the GPI was stimulated (ES = -0.02). CONCLUSIONS: In this analysis the authors highlight the need for uniform, detailed reporting of comprehensive motor and nonmotor DBS outcomes at multiple time points and for a randomized trial of bilateral STN and GPI DBS. PMID- 16381182 TI - Attenuation of fluctuating striatal synaptic dopamine levels in patients with Parkinson disease in response to subthalamic nucleus stimulation: a positron emission tomography study. AB - OBJECT: The "wearing-off" phenomenon often hampers the treatment of Parkinson disease (PD). Although deep brain stimulation (DBS) of the subthalamic nucleus (STN) is known to ameliorate the wearing-off phenomenon, the mechanism by which it does this remains unclear. As part of an inquiry into the mechanism of STN DBS, the authors measured synaptic dopamine levels in the striatum by performing positron emission tomography (PET) with [11C]raclopride. METHODS: Three patients with PD who were experiencing the wearing-off phenomenon underwent PET scanning before and after DBS of the STN. The clinical features in these patients were evaluated by applying the Hoehn and Yahr, United Parkinson's Disease Rating, and Schwab and England Activities of Daily Living Scales. Before and after surgery, PET scans were obtained using [11C]raclopride prior to and 1 hour following an oral administration of levodopa. Regions of interest for the [11C]raclopride binding potential (RacloBP) were set in the bilateral putamen and the caudate nucleus. All clinical scores were dramatically improved postoperatively. Deep brain stimulation of the STN reduced the baseline RacloBP in both the putamen and caudate nucleus, but the differences between the pre- and postoperative levels were insignificant. Before DBS of the STN, the levodopa administration significantly reduced RacloBP in the putamen (p < 0.0001). Postoperatively the drug-induced reduction in RacloBP became statistically insignificant. The drug induced increase in synaptic dopamine concentrations in the putamen preoperatively was estimated to be approximately four times higher than that after surgery (p < 0.01). The drug-induced RacloBP change in the caudate nucleus was similar to that in the putamen, although the magnitude of the change was lower (p < 0.005). The drug-induced increase in the caudate nucleus was also reduced postoperatively (p < 0.05). CONCLUSIONS: Deep brain stimulation of the STN induces the stabilization of synaptic dopamine concentrations in the striatum and may attribute to the alleviation of levodopa-related motor fluctuations. PMID- 16381183 TI - Relative importance of hypertension compared with hypervolemia for increasing cerebral oxygenation in patients with cerebral vasospasm after subarachnoid hemorrhage. AB - OBJECT: Hypervolemia and hypertension therapy is routinely used for prophylaxis and treatment of symptomatic cerebral vasospasm at many institutions. Nevertheless, there is an ongoing debate about the preferred modality (hypervolemia, hypertension, or both), the degree of therapy (moderate or aggressive), and the risk or benefit of hypervolemia, moderate hypertension, and aggressive hypertension in patients following subarachnoid hemorrhage. METHODS: Monitoring data and patient charts for 45 patients were retrospectively searched to identify periods of hypervolemia, moderate hypertension, or aggressive hypertension. Measurements of central venous pressure, fluid input, urine output, arterial blood pressure, intracranial pressure, and oxygen partial pressure (PO2) in the brain tissue were extracted from periods ranging from 1 hour to 24 hours. For these periods, the change in brain tissue PO2 and the incidence of complications were analyzed. During the 55 periods of moderate hypertension, an increase in brain tissue PO2 was found in 50 cases (90%), with complications occurring in three patients (8%). During the 25 periods of hypervolemia, an increase in brain oxygenation was found during three intervals (12%), with complications occurring in nine patients (53%). During the 10 periods of aggressive hypervolemic hypertension, an increase in brain oxygenation was found during six of the intervals (60%), with complications in five patients (50%). CONCLUSIONS: When hypervolemia treatment is applied as in this study, it may be associated with increased risks. Note, however, that further studies are needed to determine the role of this therapeutic modality in the care of patients with cerebral vasospasm. In poor-grade patients, moderate hypertension (cerebral perfusion pressure 80-120 mm Hg) in a normovolemic, hemodiluted patient is an effective method of improving cerebral oxygenation and is associated with a lower complication rate compared with hypervolemia or aggressive hypertension therapy. PMID- 16381184 TI - Prospective evaluation of surgical microscope-integrated intraoperative near infrared indocyanine green videoangiography during aneurysm surgery. AB - OBJECT: The authors prospectively compared a new technique of surgical microscope based indocyanine green (ICG) videoangiography with intraoperative or postoperative digital subtraction (DS) angiography. METHOD: The technique was performed during 187 surgical procedures in which 124 aneurysms in 114 patients were clipped. Using a newly developed setup, the ICG technique has been integrated into an operating microscope (Carl Zeiss Co., Oberkochen, Germany). A microscope-integrated light source containing infrared excitation light illuminates the operating field. The dye is injected intravenously into the patient, and intravascular fluorescence from within the blood vessels is imaged using a video camera attached to the microscope. The patency of parent, branching, and perforating arteries and documentation of clip occlusion of the aneurysm as shown by ICG videoangiography were compared with intraoperative or postoperative findings on DS angiography. The results of ICG videoangiography corresponded with intra- or postoperative DS angiography in 90% of cases. The ICG technique missed mild but hemodynamically irrelevant stenosis that was evident on DS angiography in 7.3% of cases. The ICG technique missed angiographically relevant findings in three cases (one hemodynamically relevant stenosis and two residual aneurysm necks [2.7% of cases]). In two cases the missed findings were clinically and surgically inconsequential; in the third case, a 4-mm residual neck may require a second procedure. Indocyanine green videoangiography provided significant information for the surgeon in 9% of cases, most of which led to clip correction. CONCLUSIONS: Microscope-based ICG videoangiography is simple and provides real-time information about the patency of vessels of all sizes and about the aneurysm sac. This technique may be useful during routine aneurysm surgery as an independent form of angiography or as an adjunct to intra- or postoperative DS angiography. PMID- 16381185 TI - Angiographic and clinical results in 316 coil-treated basilar artery bifurcation aneurysms. AB - OBJECT: The aim of this study was to analyze the effect of the endovascular treatment of basilar artery (BA) bifurcation aneurysms and to compare the results with those published by other neuroendovascular teams. METHODS: The authors performed a retrospective analysis of 316 aneurysms of the BA bifurcation that had been treated using endovascular coil occlusion between November 6, 1992, and February 12, 2005. After the initial embolization procedure, a 90 to 100% occlusion rate was achieved in 86% of the aneurysms. No complication was evident in 80% of the lesions, although periprocedural aneurysm rupture (3.2%) and thromboembolic events (12.3%) were the most frequent complications. Clinical outcome according to the Glasgow Outcome Scale (GOS) was a score of 5 or 4 in 77%, 3 in 11%, 2 in 5%, and 1 in 7% of patients. Initial follow-up angiography studies were obtained in 56% of patients at a mean of 19 months posttreatment and demonstrated a 90 to 100% occlusion rate in 70%. No recurrence was seen on 65% of the aneurysms. Coil compaction was evident on 24% of the follow-up angiograms. A second treatment was performed on 48 aneurysms (15%) a mean of 27 months after the first therapeutic session and resulted in 90 to 100% occlusion in 83% of the lesions. Complications were encountered in 19% of the aneurysms. Rupture did not occur during any of the procedures. Clinical outcome was rated as GOS Score 5 or 4 in 83% of the patients and Grade 3 in 17%. During a cumulative clinical follow up of 821 years in 237 patients, 182 patients (81%) were independent (GOS Score 5 or 4), 33 (14%) were dependent (GOS Score 3), eight (3%) were in a vegetative state, and two (1%) had died. Clinical outcome was significantly worse after previous aneurysm rupture and following procedural complications. CONCLUSIONS: These results are within the range of published data for coil treatment of BA tip aneurysms and confirm both the safety and efficacy of this endovascular treatment method. PMID- 16381186 TI - Modified boron neutron capture therapy for malignant gliomas performed using epithermal neutron and two boron compounds with different accumulation mechanisms: an efficacy study based on findings on neuroimages. AB - OBJECT: To improve the effectiveness of boron neutron capture therapy (BNCT) for malignant gliomas, the authors used epithermal rather than thermal neutrons for deep penetration and two boron compounds-sodium borocaptate (BSH) and boronophenylalanine (BPA)-with different accumulation mechanisms to increase the boron level in tumors while compensating for each other's faults. METHODS: Thirteen patients, 10 of whom harbored a glioblastoma multiforme (GBM), one a gliosarcoma, one an anaplastic astrocytoma, and one an anaplastic oligoastrocytoma, were treated using this modified BNCT between January 2002 and December 2003. Postoperatively, neuroimaging revealed that only one patient with a GBM had no lesion enhancement postoperatively. The patients underwent 18F-BPA positron emission tomography, if available, to assess the accumulation and distribution of BPA before neutron radiotherapy. The neutron fluence rate was estimated using the Simulation Environments for Radiotherapy Applications dose planning system before irradiation. The patients' volume assessments were performed using magnetic resonance (MR) imaging or computerized tomography (CT) scanning. Improvements in the disease as seen on neuroimages were assessed between 2 and 7 days after irradiation to determine the initial effects of BNCT; its maximal effects were also analyzed on serial neuroimages. The mean tumor volume before BNCT was 42.3 cm3. Regardless of the pre-BNCT tumor volume, in every patient harboring an assessable lesion, improvements on MR or CT images were recognized both at the initial assessment (range of volume reduction rate 17.4-71%, mean rate 46.4%) and at follow-up assessments (range of volume reduction rates 30.3-87.6%, mean rate 58.5%). More than 50% of the contrast enhanced lesions disappeared in eight of the 12 patients during the follow-up period. CONCLUSIONS: This modified BNCT produced a good improvement in malignant gliomas, as seen on neuroimages. PMID- 16381187 TI - Resection of parietal lobe gliomas: incidence and evolution of neurological deficits in 28 consecutive patients correlated to the location and morphological characteristics of the tumor. AB - OBJECT: The goal of this study is to report the incidence and clinical evolution of neurological deficits in patients who underwent resection of gliomas confined to the parietal lobe. METHODS: Patient demographics, findings of serial neurological examinations, tumor location and neuroimaging characteristics, extent of resection, and surgical outcomes were tabulated by reviewing inpatient and office records, as well as all pre- and postoperative magnetic resonance (MR) images obtained in 28 consecutive patients who underwent resection of a glial neoplasm found on imaging studies to be confined to the parietal lobe. Neurological deficits were correlated with hemispheric dominance, location of the lesion within the superior or inferior parietal lobules, subcortical extension, and involvement of the postcentral gyrus. The tumors were located in the dominant hemisphere in 18 patients (64%); had a mean diameter of 39 mm (range 14-69 mm); were isolated to the superior parietal lobule in six patients (21%) and to the inferior parietal lobule in eight patients (29%); and involved both lobules in 14 patients (50%). Gross-total resection, documented by MR imaging, was achieved in 24 patients (86%). Postoperatively, nine patients (32%) experienced new neurological deficits, whereas seven (25%) had an improvement in their preoperative deficit. A correlation was noted between larger tumors and the presence of neurological deficits both before and after resection. Postoperatively higher-level (association) parietal deficits were noted only in patients with tumors involving both the superior and inferior parietal lobules in the dominant hemisphere. At the 3-month follow-up examination, five of nine new postoperative deficits had resolved. CONCLUSIONS: Neurological deterioration and improvement occur after resection of parietal lobe gliomas. Parietal lobe association deficits, specifically the components of Gerstmann syndrome, are mostly associated with large tumors that involve both the superior and inferior parietal lobules of the dominant hemisphere. New hemineglect or sensory extinction was not noted in any patient following resection of lesions located in the nondominant hemisphere. Nevertheless, primary parietal lobe deficits (for example, a visual field loss or cortical sensory syndrome) occurred in patients regardless of hemispheric dominance. PMID- 16381188 TI - Aneurysms of the posterior circulation: detection and treatment planning using volume-rendered three-dimensional helical computerized tomography angiography. AB - OBJECT: The aim of this study was to determine whether computerized tomography (CT) angiography could be used to identify and characterize aneurysms of the posterior circulation and guide optimal treatment selection, and how data obtained using this method compared with intraoperative findings. METHODS: Patients suspected of harboring brain aneurysms underwent CT angiography and digital subtraction (DS) angiography; the results were prospectively interpreted by blinded independent evaluators. All patients with posterior circulation aneurysms were consecutively enrolled in the study. After treatment, neurosurgeons and endovascular therapists evaluated the ability of CT and DS angiography to demonstrate features of the lesions important for triage between treatment options (Wilcoxon signed-rank test) and to allow for coil or clip preselection and complete treatment planning (McNemar test of proportions), while using intraoperative findings as the basis of truth. In 242 patients overall, CT angiography detected 38 aneurysms and two aneurysmal blisters in 32 patients. The sensitivity of CT angiography in revealing posterior circulation aneurysms was 100% compared with DS angiography, with no false-positive results. Furthermore, CT angiography was sufficient as the sole study at triage for 65% of the posterior circulation aneurysms (26 of 40 lesions; p < 0.001), including 62% of the complex lesions (p < 0.001), and permitted coil or clip preselection in 74% of treated cases (20 of 27 cases; p < 0.002). Results of CT angiography revealed information about mural calcification and intraluminal thrombus not available on DS angiography, which affected patient care. CONCLUSIONS: In this study population, CT angiography was comparable to DS angiography in the detection and characterization of aneurysms of the posterior circulation. Computerized tomography angiography was used successfully to triage patients between endovascular and neurosurgical treatment options in a significant proportion of cases and permitted treatment planning in more than 70% of treated cases. PMID- 16381189 TI - Connectivity of the human periventricular-periaqueductal gray region. AB - OBJECT: The periventricular gray (PVG) zone and its continuation, the periaqueductal gray (PAG) substance, have been targets for deep brain stimulation (DBS) in the alleviation of intractable pain for longer than two decades. Nevertheless, the anatomical connectivity of this region has been fairly poorly defined. The effects of DBS in this region are probably related to the release of endogenous endorphins, but until the connectivity of this region is better understood the mechanisms will remain unclear. METHODS: Diffusion tractography was used to trace the pathways of the PVG-PAG region in seven healthy human volunteers. Images were acquired with the aid of a 1.5-tesla magnetic resonance imaging system. The region of interest was located just lateral to the posterior commissure and extended caudally to the level of the superior colliculus. Probabilistic diffusion tractography was performed from each voxel in each patient's PVG-PAG region. The PVG-PAG region was found to yield descending projections to the spinal cord and cerebellum. Ascending projections to the thalamus and frontal lobes were also observed. CONCLUSIONS: These findings suggest that the PVG-PAG region may modulate pain by two mechanisms: one involving the antinociceptive system in the spinal cord and the other involving influences on the central pain network. PMID- 16381190 TI - Local distribution and toxicity of prolonged hippocampal infusion of muscimol. AB - OBJECT: The activity of gamma-aminobutyric acid (GABA), the principal inhibitory neurotransmitter, is reduced in the hippocampus in patients with complex partial seizures from mesial temporal sclerosis. To provide preliminary safety and distribution data on using convection-enhanced delivery of agents to treat complex partial seizures and to test the efficacy and safety of regional selective neuronal suppression, the authors infused muscimol, a GABA-A receptor agonist, directly into the hippocampus of nonhuman primates using an integrated catheter electrode. METHODS: Ten rhesus monkeys were divided into three groups: 1) use of catheter electrode alone (four monkeys); 2) infusion of escalating concentrations of muscimol followed by vehicle (three monkeys); and 3) infusion of vehicle and subsequent muscimol mixed with muscimol tracer (three monkeys). Infusions were begun 5 days after catheter electrode placement and continued for 5.6 days before switching to the other agent. Head magnetic resonance (MR) images and electroencephalography recordings were obtained before and during the infusions. Brain histological studies and quantitative autoradiography were performed. Neurological function was normal in controls and when muscimol concentrations were 0.125 mM or less, whereas higher concentrations (0.5 and 1 mM) produced reversible apathy and somnolence. Fluid distribution was demonstrated on MR images and muscimol distribution was demonstrated on autoradiographs throughout the hippocampus and adjacent white matter. CONCLUSIONS: Targeted modulation of neuronal activity is a reasonable research strategy for the investigation and treatment of medically intractable epilepsy. PMID- 16381191 TI - Role of estrogen deficiency in the formation and progression of cerebral aneurysms. Part I: experimental study of the effect of oophorectomy in rats. AB - OBJECT: Estrogen has been shown to play a central role in vascular biology. Although it may exert beneficial vascular effects, its role in the pathogenesis of cerebral aneurysms remains to be determined. To elucidate the role of hormones further, the authors examined the effects of bilateral oophorectomy on the formation and progression of cerebral aneurysms in rats. METHODS: Forty-five female, 7-week-old Sprague-Dawley rats were divided into three equal groups. Group I consisted of intact rats (controls). To induce cerebral aneurysms, the animals in Groups II and III were subjected to ligation of the right common carotid and bilateral posterior renal arteries. One month later, the rats in Group II underwent bilateral oophorectomy. Three months after the experiment began all animals were killed and cerebral vascular corrosion casts were prepared and screened for cerebral aneurysms by using a scanning electron microscope. Plasma was used to determine the level of estradiol and the gelatinase activity. Hypertension developed in all rats except those in the control group. The estradiol level was significantly lower in Group II than in the other groups (p < 0.01). The incidence of cerebral aneurysm formation in Group II (60%) was three times higher than that in Group III (20%), and the mean size of aneurysms in Group II (76 +/- 27 microm, mean +/-standard deviation) was larger than that in Group III (28 +/- 4.6 microm) (p < 0.05). No aneurysm developed in control animals (Group I), and there was no significant difference in plasma gelatinase activity among the three groups. CONCLUSIONS: The cerebral aneurysm model was highly reproducible in rats. Bilateral oophorectomy increased the susceptibility of rats to aneurysm formation, indicating that hormones play a role in the pathogenesis of cerebral aneurysms. PMID- 16381192 TI - Role of estrogen deficiency in the formation and progression of cerebral aneurysms. Part II: experimental study of the effects of hormone replacement therapy in rats. AB - OBJECT: The increased incidence of cerebral aneurysms in postmenopausal women appears to be related to low levels of circulating estrogen. Using a rat model of aneurysm induction, the authors found that oophorectomy increased the incidence of experimental cerebral aneurysms (Part I in this issue). In the current study they examined the effects of hormone replacement therapy (HRT) on the formation of cerebral aneurysms in rats. METHODS: Forty-five female Sprague-Dawley rats were divided into three equal groups. The animals in Groups A and B were subjected to a cerebral aneurysm induction procedure (renal hypertension and right common carotid artery ligation) followed 1 month later by bilateral oophorectomy. After an additional week the rats in Group A received 17beta estradiol continuous-release pellets. The rats in Group C served as controls. Three months after the aneurysm induction procedure, all the rats were killed and vascular corrosion casts of their cerebral arteries were prepared and checked for aneurysmal changes. Using a scanning electron microscope, the authors recorded aneurysmal changes as endothelial changes alone (Stage I), endothelial changes with intimal pad elevation (Stage II), and saccular aneurysm formation (Stage III). Aneurysmal changes (Stages I, II, and III) occurred in one third of rats that had undergone oophorectomy and were receiving HRT (Group A), compared with 87% of the rats that had undergone oophorectomy but did not receive HRT (Group B). Although most of the aneurysmal changes identified in Group A rats were limited to Stage I or II, most changes in Group B animals were identified as saccular dilation (Stage III). CONCLUSIONS: The findings demonstrated the significant protective role of estrogen against the formation and progression of cerebral aneurysms. It appears to be related to the beneficial effects of estrogen on the function and growth of endothelial cells, which play a major role in preserving the integrity of the vascular wall. PMID- 16381193 TI - Retargeting of adenoviral vector using basic fibroblast growth factor ligand for malignant glioma gene therapy. AB - OBJECT: Adenovirus vector (AdV)-mediated gene delivery has been recently demonstrated in clinical trials as a novel potential treatment for malignant gliomas. Combined coxsackievirus B and adenovirus receptor (CAR) has been shown to function as an attachment receptor for multiple adenovirus serotypes, whereas the vitronectin integrins (alphavbeta3 and alphavbeta5) are involved in AdV internalization. In resected glioma specimens, the authors demonstrated that malignant gliomas have varying levels of CAR, alphavbeta3, and alphavbeta5 expression. METHODS: A correlation between CAR expression and the transduction efficiency of AdV carrying the green fluorescent protein in various human glioblastoma multiforme (GBM) cell lines and GBM primary cell lines was observed. To increase transgene activity in in vitro glioma cells with low or deficient levels of CAR, the authors used basic fibroblast growth factor (FGF2) as a targeting ligand to redirect adenoviral infection through its cognate receptor, FGF receptor 1 (FGFR1), which was expressed at high levels by all glioma cells. These findings were confirmed by in vivo study data demonstrating enhanced transduction efficiency of FGF2-retargeted AdV in CAR-negative intracranial gliomas compared with AdV alone, without evidence of increased angiogenesis. CONCLUSIONS: Altogether, the results demonstrated that AdV-mediated gene transfer using the FGF2/FGFR system is effective in gliomas with low or deficient levels of CAR and suggested that FGF2-retargeting of AdV may be a promising approach in glioma gene therapy. PMID- 16381194 TI - Nerve regeneration across a 2-cm gap in the rat median nerve using a resorbable nerve conduit filled with Schwann cells. AB - OBJECT: In a rat model, nerve regeneration was evaluated across a 2-cm defect in the median nerve by using a resorbable artificial nerve conduit. The aim of this study was to develop an artificial, biocompatible nerve guide to induce regeneration in the peripheral nervous system. METHODS: The authors compared a nerve conduit of trimethylenecarbonate-co-epsilon-caprolactone (TMC/CL) filled with autologous Schwann cells with both an empty hollow conduit and an autologous nerve graft. Animals that did not undergo surgery served as the control group. Nerve regeneration was evaluated with the grasping test, histological analysis of the nerve, muscle weight analysis (flexor digitorum superficialis muscle), and electrophysiological examination. After an observation period of 9 months, regeneration occurred only in animals that had received an autologous graft or a Schwann cell containing nerve conduit. No signs of regeneration were found in animals supplied with the empty conduit. CONCLUSIONS: Results of this study reveal the important role of Schwann cells in the regeneration process across a 2 cm defect in the rat median nerve. Furthermore, Schwann cell-filled nerve conduits induced functional recovery, as demonstrated in the grasping test, that was comparable with that of the autologous graft 9 months after implantation. PMID- 16381195 TI - Synovial sarcoma of the frontal sinus. Case report. AB - Synovial sarcoma is a soft-tissue lesion occurring predominantly in the extremities of young adults. Although the head and neck region is the second most common site of involvement, synovial sarcoma has rarely been reported in the paranasal sinus. The authors present a case of synovial sarcoma arising from the frontal sinus and review the literature of synovial sarcomas arising from the paranasal sinuses. PMID- 16381196 TI - Camptocormia in a patient with Parkinson disease: beneficial effects of pallidal deep brain stimulation. Case report. AB - Camptocormia is a well-known feature in cases of Parkinson disease (PD), and is usually unresponsive to dopaminergic therapy. A 62-year old man with a long history of PD and severe progressive camptocormia underwent bilateral pallidal stimulation. During the next 6 months a sustained improvement in trunk flexion was observed. After 14 months of follow up, the patient's trunk flexion is almost normal. PMID- 16381197 TI - Rapid expansion of a previously asymptomatic subependymoma. Case report. AB - This 39-year-old man presented with a 6-month history of occipital headaches. Magnetic resonance imaging revealed an irregularly shaped fourth ventricle mass. One month after his initial presentation, he was admitted to the hospital with significant tumor expansion and clinical deterioration. A posterior fossa craniectomy was performed and the mass was resected. Histopathological analysis of this tumor showed central necrosis with associated edema in an otherwise typical and benign-appearing subependymoma. To the authors' knowledge, this is the first reported case of rapid, nonhemorrhagic expansion associated with necrosis in a previously asymptomatic subependymoma. PMID- 16381198 TI - Arachnoid cyst with associated arachnoiditis developing after subarachnoid hemorrhage. Case report. AB - The authors report the case of a 53-year-old woman in whom a T1-T2 spinal arachnoid cyst with associated arachnoiditis developed, compressing the thoracic spinal cord 1 year after the patient had suffered a Hunt and Hess Grade IV subarachnoid hemorrhage (SAH). Development of spinal arachnoiditis with or without an arachnoid cyst is a rare complication of aneurysmal SAH. Risk factors may include posterior circulation aneurysms, the extent and severity of the hemorrhage, and the need for cerebrospinal fluid diversion. Surgical drainage, shunt placement, or cyst excision, when possible, is the mainstay of treatment. PMID- 16381199 TI - Pituitary cyst presenting with hyponatremia and increased secretion of brain natriuretic peptide. Case report. AB - In most cases of pituitary cyst there are no clinical symptoms and the lesions are found incidentally. The authors report the case of a 60-year-old man with a pituitary cyst causing visual disturbance and hyponatremia. The patient presented with appetite loss and general fatigue. On admission, blood workup showed severe hyponatremia (112 mEq/L), and bitemporal hemianopsia was observed on neurological examination. Magnetic resonance imaging revealed an intra- and suprasellar region cystic mass extending to the frontal base and hypothalamic area. The serum level of brain natriuretic peptide (BNP) was elevated (92 pg/ml) with polyuria and excessive Na excretion. Transsphenoidal surgery was performed to drain the cyst. The cyst wall was partially excised and the cystic fluid was aspirated. The secretion of BNP normalized postoperatively, and the hyponatremia and visual symptoms resolved. Histological examination, including an electron microscopy study, confirmed the diagnosis of a simple cyst. This appears to be the first reported case of a pituitary simple cyst associated with hyponatremia and an elevated BNP level. PMID- 16381200 TI - Rathke cleft cyst of the sphenoid bone and nasopharynx. Case illustration. PMID- 16381201 TI - Harvey Cushing and Oskar Hirsch: early forefathers of modern transsphenoidal surgery. AB - The transnasal transsphenoidal approach is the preferred route for removal of most lesions of the sella turcica. The concept of transnasal surgery traversing the sphenoid sinus to reach the sella has existed for nearly a century. A comprehensive historical overview of the evolution of transsphenoidal surgery has been reported previously. In the present vignette, the authors focus on transsphenoidal surgery in the early 1900s, particularly on the methods advocated by Harvey Cushing and Oskar Hirsch, two prominent pituitary surgeons who pioneered the transsphenoidal technique. Cushing championed the sublabial approach, whereas Hirsch was the master of the endonasal route. Coincidentally, both surgeons independently performed the submucous septal resection for the first time on June 4, 1910. Although Cushing's and Hirsch's approaches were predicated on the work of their predecessors, their transsphenoidal procedures became the two most popular techniques and, for future generations of pituitary surgeons, laid the foundation for modem transsphenoidal surgery. In this comparative analysis, the authors compare the operative nuances of the approaches of Cushing and Hirsch and describe the contributions of these pioneers to modern transsphenoidal surgery. PMID- 16381202 TI - Assessment of adjacent-segment disease in patients treated with cervical fusion or arthroplasty: a prospective 2-year study. AB - OBJECT: The authors compared the incidence of radiologically documented changes and symptomatic adjacent-level cervical disc disease after single-level discectomy and subsequent cervical fusion or arthroplasty in two independent prospective clinical studies. METHODS: The patients were treated with the Affinity Anterior Cervical Cage System or the Bryan Artificial Cervical Disc. In each study the patients were required to undergo serial cervical radiography preoperatively and 24 months postoperatively, as well as serial clinical evaluations including documentation of adverse events, neurological status, and results of the 36-item Short Form Health Survey. All serial radiographs were reviewed prior to evaluating the clinical symptoms for development of increasing or new adjacent degenerative disc disease (DDD). Subsequently, the clinical data were analyzed. For various reasons of exclusion, the cases analyzed in the Bryan disc-treated cohort consisted of 74 patients and in the Affinity system-treated cohort there were 158 patients. New anterior osteophyte formation or enlargement, increased narrowing of an interspace, new DDD, and calcification of the anterior longitudinal ligament were the radiological findings indicative of adjacent-level disease. Fusion was associated with a significant increase in x-ray film-based changes of adjacent-disc disease (p = 0.009, odds ratio [OR] 2.44). In the cage fusion series, the incidence of symptomatic adjacent-level DDD was statistically greater than that in the group treated with the artificial disc (p = 0.018), and the patients required a statistically greater number of medical treatments related to episodic symptoms of neck, shoulder, and arm pain attributed to new disc disease (p = 0.001, OR 35.8). CONCLUSIONS: In comparing these prospective studies the authors demonstrated that maintaining motion rather than fusion will prevent symptomatic adjacent-disc disease and will decrease adjacent-level radiological indicators of disease at a 24-month postoperative interval. PMID- 16381203 TI - Cervical disc arthroplasty: a controlled randomized prospective study with intermediate follow-up results. Invited submission from the joint section meeting on disorders of the spine and peripheral nerves, March 2005. AB - OBJECT: The authors of studies of anterior cervical fusion for disc-related disease have documented satisfying clinical results and infrequent complications. Recently, cervical disc arthroplasty has entered clinical trials as an alternative to fusion. Although the results of anecdotal reports and prospective studies support this procedure, these studies have not conferred the validity of a controlled randomized study. In the present study, the author presents data obtained in such a study. METHODS: After symptoms failed to respond to conservative therapy, 46 patients with one-level discogenic cervical radiculopathy and/or myelopathy were randomized to undergo arthroplasty or fusion as part of a US Food and Drug Administration medical device study. In all patients there was a minimum follow-up duration of 1 year. Equivalent results for the relief of arm pain and neck pain were noted in both treatment groups. Treatment parameters other than operative time were similar. No neurological or serious systemic complications occurred. CONCLUSIONS: Although extended follow-up data and larger patient populations are needed, the results of this study indicate that arthroplasty is a viable alternative to cervical fusion. PMID- 16381204 TI - Primary spinal syringomyelia. Invited submission from the joint section meeting on disorders of the spine and peripheral nerves, March 2005. AB - In the present review the author describes the different types of syringomyelia that originate from abnormalities at the level of the spinal cord rather than at the craniovertebral junction. These include posttraumatic and postinflammatory syringomyelia, as well as syringomyelia associated with arachnoid cysts and spinal cord tumors. The diagnosis and the principles of managing these lesions are discussed, notably resection of the entity restricting cerebrospinal fluid flow. Placement of a shunt into the syrinx cavity is reserved for patients in whom other procedures have failed or who are not candidates for other procedures. PMID- 16381205 TI - Safety of transforaminal lumbar interbody fusion and intervertebral recombinant human bone morphogenetic protein-2. AB - OBJECT: Recombinant human bone morphogenetic protein-2 (rhBMP-2) is being increasingly used for spinal fusion. There are few data regarding its clinical safety, effectiveness, and clinical outcome when applied on an absorbable collagen sponge (ACS) in conjunction with allograft for transforaminal lumbar interbody fusion (TLIF). METHODS: Seventy-four consecutive patients undergoing TLIF for degenerative disc disease were divided into five groups depending on whether the patient underwent a minimally invasive or open approach, as well as the number of spinal levels surgically treated. Surgery-related data, fusion results, complications, and clinical outcome were evaluated. The mean follow-up duration was 20.6 months (range 14-28 months). The radiographic fusion rate was 100% at 12 and 24 months after the surgery. No bone overgrowth or other complications related to BMP use were demonstrated. CONCLUSIONS: Analysis of the results demonstrated that TLIF combined with a BMP-2-soaked ACS is a feasible, effective, and safe method to promote lumbar fusion. There were no significant intergroup differences in clinical outcome between patients who underwent open compared with minimally invasive procedures. Patient satisfaction rates, however, were higher in the minimally invasive procedure group. The efficacy of BMP-2 was not dependent on which approach was used or the number of spinal levels that were treated. PMID- 16381206 TI - Computerized tomography-guided kryorhizotomy in 76 patients with lumbar facet joint syndrome. AB - OBJECT: The authors prospectively evaluated the therapeutic effect of computerized tomography (CT)-guided kryorhizotomy in the treatment of patients with lumbar facet joint syndrome (LFJS) and assessed prognostic factors that predict this effect. METHODS: Between February 2001 and March 2004, CT-guided kryorhizotomy of facet joints was performed in 76 patients with LFJS. A diagnosis was established after three positive CT-guided medial nerve branch blocks. Outcome was determined by evaluating the results of a standardized questionnaire, including visual analog scale (VAS) score, use of medication, ability to work, and physical conditions. Measurement was performed before treatment and repeated postoperatively at 3 days, 3 months, and every 6 months thereafter. On September 2004 all patients underwent clinical reevaluation. The median follow-up period was 22.5 months (range 6-43 months); the median interval to pain reduction was 6 months (range 0.1-31 months) after the first kryorhizotomy. The mean VAS pain score was 6.7 preoperatively and 2.9, 3.2, and 3.4 at 3 days, 3 months, and 6 months postoperatively, respectively. In 40% of patients pain was reduced for 12 months or longer. In patients in whom there was no prior surgical treatment of the relevant spinal segment, the duration of pain relief was significantly longer than in patients who had previously undergone surgery (p < 0.03). Eighteen patients underwent a second, seven a third, and one a fourth kryorhizotomy. No patient reported any side effect. The use of CT guidance guarantees an exact needle-tip position control and documentation for repeated procedures. CONCLUSIONS: Computerized tomography-guided kryorhizotomy is a minimally invasive and repeatable treatment that yields good long-term results in patients with LFJS. PMID- 16381207 TI - Thoracic and lumbar tuberculous spondylitis treated by posterior debridement, graft placement, and instrumentation: a retrospective analysis in 19 cases. AB - OBJECT: Surgical treatment of thoracic and lumbar tuberculous spondylitis is controversial. An anterior approach is usually recommended. The aim of the present study was to assess the efficacy of posterior debridement and the placement of posterior instrumentation for the treatment of patients with thoracic and lumbar tuberculous spondylitis. METHODS: Nineteen patients with thoracic and lumbar tuberculous spondylitis underwent single-stage posterior decompression and debridement as well as the placement of posterior interbody grafts if necessary, instrumentation and posterior or posterolateral grafts. No postoperative neurological deterioration was noted. One patient died of myocardial infarction on Day 10. The mean follow-up duration, excluding the one death, was 52.7 months (range 16-125 months). In a 70-year-old patient, a single pedicle screw broke after 3 months. All patients were in better neurological condition after surgery and at the last follow-up examination. Neurological deficits were present in only two patients at the last follow up (one American Spinal Injury Association Grade B and one Grade C deficit preoperatively). Three other patients suffered intermittent back or low-back pain. The mean angulation measured in 13 patients with kyphotic deformity was 18.2 degrees (range 5-42 degrees) preoperatively; this was reduced to 17.3 degrees (range 0-42 degrees) after surgery. There was a 2.8 degrees loss of correction (range 2-5 degrees) after 44.3 months (16-64 months). Kyphosis did not progress beyond 15 months in any patient. CONCLUSIONS: A posterior approach in combination with internal fixation and posterior or posterolateral fusion (with or without placement of posterior interbody grafts) may be sufficient for the debridement of the infection and to allow spinal stabilization in patients with thoracic and lumbar tuberculous spondylitis. This procedure is associated with easy access to the spinal canal for neural decompression, prevention of loss of corrected vertebral alignment in the long term, and facilitation of early mobilization. PMID- 16381208 TI - Endoscopic thoracic microdiscectomy. AB - OBJECT: The purpose of this clinical study was to evaluate prospectively surgical and neurological outcomes after endoscopic thoracic disc surgery. METHODS: The authors assessed the following quantifiable outcome data in 46 patients: operative time, blood loss, duration of chest tube insertion, narcotic use, hospital length of stay (LOS), and long-term follow-up neurological function and pain-related symptoms. In patients who presented with myelopathy there was a postoperative improvement of two Frankel grades. Pain related to radiculopathy was improved by 75% and in one patient it worsened postoperatively. The authors also present operative data, surgical outcomes, and complications. CONCLUSIONS: Thoracoscopic discectomy can be used to achieve acceptable results. It has several distinct advantages such as reduced postoperative pain, morbidity, and LOS compared with traditional open procedures. PMID- 16381209 TI - Biomechanical analysis of a newly designed bioabsorbable anterior cervical plate. Invited submission from the joint section meeting on disorders of the spine and peripheral nerves, March 2005. AB - OBJECT: The authors present a biomechanical analysis of a newly designed bioabsorbable anterior cervical plate (ACP) for the treatment of one-level cervical degenerative disc disease. They studied anterior cervical discectomy and fusion (ACDF) in a human cadaveric model, comparing the stability of the cervical spine after placement of the bioabsorbable fusion plate, a bioabsorbable mesh, and a more traditional metallic ACP. METHODS: Seven human cadaveric specimens underwent a C6-7 fibular graft-assisted ACDF placement. A one-level resorbable ACP was then placed and secured with bioabsorbable screws. Flexibility testing was performed on both intact and instrumented specimens using a servohydraulic system to create flexion-extension, lateral bending, and axial rotation motions. After data analysis, three parameters were calculated: angular range of motion, lax zone, and stiff zone. The results were compared with those obtained in a previous study of a resorbable fusion mesh and with those acquired using metallic fusion ACPs. For all parameters studied, the resorbable plate consistently conferred greater stability than the resorbable mesh. Moreover, it offered comparable stability with that of metallic fusion ACPs. CONCLUSIONS: Bioabsorbable plates provide better stability than resorbable mesh. Although the results of this study do not necessarily indicate that a resorbable plate confers equivalent stability to a metal plate, the resorbable ACP certainly yielded better results than the resorbable mesh. Bioabsorbable fusion ACPs should therefore be considered as alternatives to metal plates when a graft containment device is required. PMID- 16381210 TI - Localized cervical facet joint kinematics under physiological and whiplash loading. AB - OBJECT: Although facet joints have been implicated in the whiplash injury mechanism, no investigators have determined the degree to which joint motions in whiplash are nonphysiological. The purpose of this investigation was to quantify the correlation between facet joint and segmental motions under physiological and whiplash loading. METHODS: Human cadaveric cervical spine specimens were exercise tested under physiological extension loading, and intact human head-neck complexes were exercise tested under whiplash loading to correlate the localized component motions of the C4-5 facet joint with segmental extension. Facet joint shear and distraction kinematics demonstrated a linear correlation with segmental extension under both loading modes. Facet joints responded differently to whiplash and physiological loading, with significantly increased kinematics for the same-segmental angulation. The limitations of this study include removal of superficial musculature and the limited sample size for physiological testing. CONCLUSIONS: The presence of increased facet joint motions indicated that synovial joint soft-tissue components (that is, synovial membrane and capsular ligament) sustain increased distortion that may subject these tissues to a greater likelihood of injury. This finding is supported by clinical investigations in which lower cervical facet joint injury resulted in similar pain patterns due to the most commonly reported whiplash symptoms. PMID- 16381211 TI - Gowers intrasyringal hemorrhage. Case report and review of the literature. AB - The concept of hemorrhage in a preexisting syringomyelic cavity was first described by Gowers in 1904. Since its first description only 13 cases have been reported. The aims of this report are to describe a new case, bring this entity to wider attention, and summarize the existing literature on the subject. This 36 year-old woman presented with progressive gait disturbance and unsteadiness. Physical examination revealed incomplete quadriparesis, predominantly on the left side, and hypesthesia below C-7. Magnetic resonance imaging revealed hematomyelia characterized by a heterogeneous hyperintense signal within the central cervical cord. A liquefied well-limited hematoma was evacuated. The postoperative course was uneventful; a near-complete recovery was observed at the 7-year follow-up examination. Most cases of intrasyringal hemorrhage (ISH) have occurred in syringomyelic cavities associated with scoliosis or Chiari malformation Type I. Although there is no specific clinical picture associated with this entity, it can be characterized by three neurological forms: 1) sudden onset or rapid development of signs and symptoms, 2) acute worsening of symptoms that may improve but leaving greater neurological dysfunction than before the previous episode, and 3) ISH may initiate progressive deterioration in a patient with known syringomyelia. Intrasyringal bleeding is most probably caused by a sudden dilation of the syringomyelic cavity, which may provoke rupture of the intrasyringal vessels by an acute distension of the accompanying strands. Magnetic resonance imaging is the most accurate diagnostic modality, and recognition of ISH can lead to early, safe, and efficient surgical treatment. PMID- 16381212 TI - An unusual presentation and resolution of syringomyelia after cervical spine injury. Case report. AB - The authors describe a case of noncommunicating syringomyelia associated with Chiari malformation Type I in a patient in whom acute symptomatic exacerbation occurred following cervical spine trauma. Surgical stabilization and realignment of the spine resulted in marked resolution of the neurological abnormalities, and subsequent magnetic resonance imaging demonstrated persistent collapse of the syrinx. The authors review the various factors in the pathogenesis of this unusual sequence of events. PMID- 16381213 TI - A large, erosive intraspinal and paravertebral gout tophus. Case report. AB - Symptomatic gout tophi of the spine are a rare but well-characterized complication of tophaceous gout. The authors report the case of a 29-year-old previously healthy man who presented with L-5 radiculopathy. Lumbar magnetic resonance (MR) imaging revealed a 4.5 x 4.5 x 2.8-cm large gout tophus mimicking a malignant spinal tumor or abscess. The tophus completely destroyed both L-4 and L-5 facet joints and the left L-4 lamina and spread epidurally from L-3 to L-5, compressing the left L-5 nerve root. There has been no similar case reported so far with respect to the extent of bone destruction. The authors describe the case history and present intraoperative, MR imaging, and histological findings. PMID- 16381214 TI - Spinal intradural arteriovenous fistulas acquired in late adulthood: absent spinal venous drainage in pathogenesis and pathophysiology. Report of two cases. AB - Intradural spinal arteriovenous fistulas (AVFs), a subtype of spinal arteriovenous malformation in which there is a direct communication between a spinal artery and a vein on the cord surface or in the subarachnoid space, are generally considered to be congenital lesions caused by maldevelopment of the embryonic vascular system. The authors present the cases of two patients with acquired AVFs of the terminal filum. In each patient an AVF between the distal segment of the anterior spinal artery and its accompanying vein on the terminal filum developed within 1 year of repeated lumbar myelography that had demonstrated no evidence of abnormal vascularity. In both patients spinal arteriography demonstrated the absence of medullary venous drainage in the thoracolumbar region, which, combined with the arterialized venous input from the AVF, permitted the development of venous congestion and myelopathy. The involved segment of the terminal filum was excised; in vitro microarteriography and the histopathological examination demonstrated a single, simple arteriovenous connection in both patients. The findings in these cases indicate that intradural AVF can spontaneously arise in later life. The development of these lesions and/or their clinical manifestation may require not only the presence of the AVF, but also deficiency of medullary spinal venous drainage. The epidemiology and anatomy of intradural AVFs are compatible with an acquired origin in many cases. PMID- 16381215 TI - Spinal hemangioblastoma containing metastatic renal cell carcinoma in von Hippel Lindau disease. Case report and review of the literature. AB - The authors describe the case of a patient with von Hippel-Lindau (VHL) disease in which a spinal hemangioblastoma contained metastatic renal cell carcinoma (RCC). The literature on tumor-to-tumor metastasis associated with VHL disease of the central nervous system (CNS) is reviewed. Midthoracic back pain developed in this 43-year-old man with a left-sided radicular component 2 years after he underwent resection of a left RCC. Radiological findings demonstrated a T6-7 intradural intramedullary lesion. A T5-8 laminectomy and gross-total resection of the spinal cord mass were performed. Light and electron microscopic examination showed features of hemangioblastoma, which contained metastatic foci of RCC. Genetic analysis demonstrated the presence of a deleting mutation in the first exon (nt. 394-406) of the VHL locus, truncating 16 amino acids (N61-77) from the first beta sheet in the VHL protein. A review of the literature revealed that RCC to-CNS hemangioblastoma is the second most common donor-recipient tumor association among the tumor-to-tumor metastases. PMID- 16381216 TI - Spinal pelvic reconstruction after total sacrectomy for en bloc resection of a giant sacral chordoma. Technical note. AB - Although radical resection prolongs the disease-free survival period, surgical management of primary sacral tumors is challenging because of their location and often large size. Moreover, in cases of lesions for which a radical resection necessitates total sacrectomy, reconstruction is required. The authors have previously described a modified Galveston technique in which a liaison between the spine and pelvis is achieved using lumbar pedicle screws and Galveston rods embedded into the ilia; additionally, a transiliac bar reestablishes the pelvic ring. Although this reconstruction technique achieves stabilization, several biomechanical limitations exist. In the present report the authors present the case of a patient who underwent spinal pelvic reconstruction after a total sacrectomy was performed to remove a giant sacral chordoma. They describe a novel spinal pelvic reconstruction technique that addresses some of the biomechanical limitations. PMID- 16381217 TI - Age and lumbar surgery. PMID- 16381218 TI - Spinal cord herniation. PMID- 16381219 TI - Atlantoaxial screw fixation. PMID- 16381221 TI - Spotlight on success. Val Doyle. PMID- 16381222 TI - Career pathways in sexual health. PMID- 16381223 TI - Implementing change in sexual health care. PMID- 16381224 TI - Strategic thinking. PMID- 16381225 TI - Targeting sexually transmitted infections. AB - Nurses and other healthcare professionals working in the field of sexual health face significant challenges. The way that resources, which are allocated on the basis of the Choosing Health consultation, are used will have an impact on service modernisation and new opportunities for service delivery. But targets and the desire among PCTs to meet their LDPs must not compromise clinical quality, and robust mechanisms for clinical governance must be set in place. These should include: The education and training of all healthcare professionals, The development of effective pathways for referral to specialist GUM providers, Arrangements for ongoing supervision and support of clinical work undertaken in non GUM settings. For many patients, access to services and plurality of supply are of paramount importance. New technology is making different patterns of service delivery possible, as is better use and development of staff, particularly nurses. Nurses however must decide upon and, if necessary, develop nationally recognised specialist education in STIs that supports the development of skills and expertise from novice to expert. Whether this education should, and could, be delivered uni- or multiprofessionally is a matter for ongoing debate. What must be clarified however are the levels of competence achieved and their parity across professional groups. Ultimately, whether government targets are achievable remains to be seen. What is important is the fact that, by setting targets, the government has finally recognised STIs and GUM as health priorities. PMID- 16381226 TI - In strictest confidence. PMID- 16381227 TI - Scope of practice. PMID- 16381228 TI - Overseas recruitment and migration. PMID- 16381229 TI - [Viral hepatitis: Croatian consensus statement]. AB - There has been a dramatic improvement in diagnostic procedures and therapy of viral hepatitis in the last 20 years. Improvements in therapy caused an increase in actual cost, however, with significant long-term savings through a decreased cost of treatment of advanced liver disease including liver transplantation. The Croatian National Board for Viral Hepatitis has decided to initiate the organization of consensus conference on viral hepatitis enabling the leading experts in the country to give the best possible recommendations for the diagnosis, prophylaxis and therapy in our circumstances. The Consensus Conference took place in Zagreb in June 2004, with update in March 2005, organized by the Croatian National Board for Viral Hepatitis, Reference Centers of the Ministry of Health for Chronic Liver Diseases, Infectious Diseases and AIDS, Croatian Society of Gastroenterology--Hepatology Section, Croatian Society for Nephrology, Dialysis and Transplantation, and Croatian Institute for Health Insurance. Invited experts provided written reports on the respective subjects that appear in this issue and their recommendations resulting in this consensus statement. PMID- 16381230 TI - [Epidemiology of hepatitis B and C in Croatia]. AB - The incidence of HBV and HCV infection is hard to determine because of the high number of asymptomatic infections. According to data of the Croatian Institute of Public Health, there are 200 newly infected persons with hepatitis B and approximately the same number of newly identified HBsAg carriers occur each year. Accordingly, Croatia is among the countries with less than 2% of HBsAg carriers in the general population. In these circumstances, HBV infection is most often spread among adolescents and younger adults. The route of transmission is most often sexual (semen) or through the skin in high-risk groups. An increased risk of infection is found in newborns of HBsAg positive mothers, i.v. addicts, promiscuous individuals, male homosexuals, person in close contact with acutely ill or chronic HBsAg carriers, persons that come in contact with blood and other potentially contaminated body fluids, dialysis patients, patients with multiple blood transfusions, patients with transplanted organ or tissue, patients treated for hematologic malignancies and hemophilia, and persons who undergo acupuncture, tattooing or piercing, or travel to areas with a high prevalence of HBV infection. The estimated prevalence of HCV infection marker (anti-HCV) in the Croatian general population is more than 1% and the number of yearly infected with hepatitis C reported to the Croatian Institute of Public Health is around 200 cases. The highest incidence is found in the 20-40 age groups at a high risk of infection by the use of drug injection. At risk are persons who received transfusion of blood or blood products prior to the availability of blood screening of voluntary blood donors. PMID- 16381231 TI - [Prevention of hepatitis B and C]. AB - Preventive measures are determined by epidemiological characteristics of HBV and HCV infections. A significant difference between these two infections is in the availability of effective vaccine for hepatitis B. Vaccine against hepatitis C is not available. The vaccination program for hepatitis B in Croatia was initiated in 1994, at first only for high risk groups. The vaccination program for children in the 6th grade of primary school started in 1999. Besides that, all pregnant women are obliged to undergo HBV testing. HBV infection is very rare but not unknown in early childhood. HBV infection in early childhood, most commonly asymptomatic, results in very high prevalence of chronic infection and carries a high risk of late serious consequences of the HBV infection. Since the main aim of vaccination is to prevent serious chronic liver disease, it is very important to protect the youngest children. The prevention of hepatitis C consists of primary prevention, mostly eliminating the risks of the presence and spread of infection. It is recommended to follow instructions regarding prevention of transmission by skin and mucosal contact with blood and body fluids, especially in health institutions and during hemodialysis. Providing due education to high risk persons (i.v. addicts, persons with more than one sexual partner, patients and workers who come in contact with blood and other body fluids) is very important. Early identification of infected persons enables timely treatment. It is not recommended to screen general population but only persons at high risk identified by medical history and other relevant medical information. PMID- 16381232 TI - [Hepatitis B and C: natural course of disease]. AB - Significant progress in the understanding of the natural history of hepatitis B and C has been made in recent years due to molecular diagnosis techniques. The most important biologic feature of hepatitis B and C viruses (HBV, HCV) is their ability to cause chronic hepatitis. The natural course of HBV infection is variable, ranging from inactive HBsAg carrier state to progressive chronic hepatitis that can evolve into liver cirrhosis and hepatocellular carcinoma. HBeAg-negative chronic hepatitis is due to a naturally occurring HBV variant with mutations in the precore or basic core promoter regions. It accounts for the majority of cases in many European countries and is generally associated with a more severe liver disease. The morbidity and mortality in chronic hepatitis B are linked to the evolution to cirrhosis and hepatocellular carcinoma. The progression of fibrosis is strongly associated with persistent active viral replication When the diagnosis is made, the 5-year cumulative incidence of developing cirrhosis ranges from 8% to 20%. The 5-year cumulative incidence of hepatic decompensation is 20%. Hepatocellular carcinoma is one of the most common cancers worldwide, 75% of which are related to chronic HBV infection. Coinfection with hepatitis D virus can lead to a more progressive liver disease in a shorter period of time. Hepatitis C virus infection becomes chronic in 80% of infected persons resulting in different stages of chronic hepatitis, with 20%-30% progressing to cirrhosis within 20 years period. The progression of fibrosis determines the ultimate prognosis. The major factors known to be associated with fibrosis progression are older age, male gender and alcohol consumption. Viral load and genotype do not play a role in the disease progression. Progression to fibrosis is more rapid in immunocompromised patients. PMID- 16381233 TI - [Factors influencing clinical course of viral hepatitis]. AB - Hepatitis B and C are diseases characterized by a high global prevalence, complex clinical course and limited efficacy of currently available antiviral therapy. Hepatitis B: local factors have a significant influence not only on the disease prevalence but also on the disease course. Vertical transmission of the infection in the areas of high prevalence results in perinatal infection, which universaly leads to the development of chronic disease. Factors associated with an increased risk of cirrhosis are older age, persistent viremia, coinfection with HCV, HDV and HIV, and consumption of alcohol, while the role of viral genotype is uncertain. Predictors of HCC development in cirrhotic liver are older age, male sex, alcohol abuse, exposure to aflatoxin, coinfection with HCV and HDV, continuously active inflammation, and potentially viral genotype. Survival predictors in cirrhotic patients are age, serum albumin, platelet count and splenomegaly as a reflection of portal hypertension. Hepatitis C: the risk of cirrhosis is low. Risk factors for cirrhosis are infection in older age, alcohol abuse, and coinfection with HBV and HIV. Obesity has negative impact on treatment efficacy. PMID- 16381234 TI - [Viral hepatitis: clinical and histological assessment]. AB - Viral factors are important in the pathogenesis of liver disease either directly, through cell injury associated with the accumulation of intact virus or viral proteins, or indirectly, through the host immune response. The immune response of the host is more important than viral factors in the pathogenesis of liver injury caused by HBV. There is little genomic variability in HBV, but mutant forms of HBV with mutations in the precore, surface and X genes, as well as the core promoter region have been implicated in a number of clinical syndromes. The mechanisms of viral persistence and hepatocellular injury in patients with hepatitis C infection are poorly understood. Viral infection can produce cellular injury by direct cytotoxicity and by indirect immune-mediated injury. The range of histologic findings in patients with hepatitis B and C is broad, from minimal inflammatory changes to fulminant hepatitis, liver cirrhosis and hepatocellular carcinoma. The goal of histologic analysis is quantitative and qualitative assessment of necroinflammatory lesions and fibrosis, changes of liver structure and follow-up of histologic changes due to the natural course of the disease or antiviral therapy. Subjective impression in the assessment of histologic changes is not satisfactory and numerical indices should be used. Presently, Knodell's index is being replaced by METAVIR system and Ishak system. Clinical assessment of patients with viral hepatitis must include detailed physical examination, description of the clinical course and extrahepatic manifestations, definition of viral factors, histologic assessment of inflammatory activity and degree of fibrosis, and a number of relevant laboratory parameters. PMID- 16381235 TI - [Laboratory diagnosis of viral hepatitis B and C]. AB - Accurate diagnosis of viral hepatitis is based on determination of specific viral markers. In HBV infection they include HBsAg, anti-HBs, HBeAg, anti-HBe, anti HBc, IgM anti-HBc, and HBV DNA. There are patients with HBV marker constellation indicating serologic recovery, but with HBV DNA in the liver indicating continuous viral replication. Mutations have been described in all four HBV genes. It is important to take into account the main precore mutation which leads to a decrease or complete inhibition of HBeAg production (HBeAg negative HBV infection). Diagnostically most important are surface gene mutations because they can result in the false diagnosis or delay in diagnosis in important groups of patients. Anti-HCV and HCV RNA are found in sera of patients with HCV infection. A false positive result is possible with ELISA, especially in patients with low c/o ratio and in all individuals with a low risk of HCV infection. It is necessary to confirm ELISA positivity with confirmation techniques (western blot, immunoblot). There are qualitative and quantitative assays for HCV RNA determination. HCV genotyping should be done, since different viral genotypes respond differently to therapy and therapeutic protocols are different. It is possible to determine HCVAg free or complexed with the antibody. Determination of free HCVAg could enable the diagnosis of acute HCV infection. There are some clinical situations where the interpretation of HBV and HCV markers is difficult because of ambiguous interpretation and "unusual" constellation. Attention should be focused on simultaneous infection within other hepatitis viruses or other viruses like HIV. Replication of one virus could have a direct influence on the replication and expression of another virus. PMID- 16381236 TI - [Monitoring patients with chronic hepatitis during and after therapy]. AB - Hepatitis C is a major public health problem. General screening is not advisable and should be limited to risk groups. The gold standard for the assessment of disease severity is liver biopsy. AST and ALT do not correlate with histology. Serum HCV RNA by qualitative assay and HCV genotype should be determined prior to therapy. Response to antiviral therapy should be assessed by testing AST, ALT and qualitative HCV RNA. Repeat liver biopsy is not necessary. The incidence of HCC related to HCV infection is rising. Early detection by a cost effective screening program is essential. In patients with liver cirrhosis caused by hepatitis C, alpha fetoprotein and liver sonography should be done every 6 months. Upper GI endoscopy is recommended every 1-4 years in cirrhotic patients. Over 350 000 000 people are infected with HBV worldwide, and chronic HBV infection is the leading cause of liver cancer and tenth leading cause of death. HBs Ag, HBeAg and HBV DNA positive patients should be monitored for 6 months before treatment. Patients treated with antiviral therapy should be tested for HBAg, HBeAg and HBV DNA at the end of treatment and every 6 months thereafter to assess virologic response. Monitoring of serum HBV DNA is done by PCR. Patients treated with lamivudine should be tested for YMDD mutation. Ultrasound and AFP monitoring are recommended for detection of HCC, but results are not always reliable. Approximately 40% -70% of HIV infected patients have coinfection with HCV, HBV and HDV. HIV/HCV coinfected patients have an increased risk of progressive liver disease and should be treated accordingly. PMID- 16381237 TI - [Hepatitis B: who should be treated?]. AB - Patients with moderate to severe chronic hepatitis and active HBV replication should be managed according to HBeAg status and presence of coinfecting virus(es) (HDV, HCV, HIV). Treatment is also recommended for immunocompromised patients and those older than 60 years with acute HBV infection. Current treatment of chronic hepatitis B has limited long-term efficacy. The patient's age, severity of liver disease, likelihood of response and possibility of adverse effects and complications should be considered before deciding on treatment. PMID- 16381238 TI - [Hepatitis C: who should be treated?]. AB - The goals of treatment for hepatitis C are eradication of viral infection and prevention of progression to end-stage liver disease. To decide on whom to treat, it is necessary to consider the following parameters: stage of disease, comorbidity, patient age, probability of therapeutic success, contraindications, and therapy associated risks. Therapy is recommended in patients with elevated ALT levels, patients with normal ALT level and at least F2 METAVIR score, nonresponders to standard interferon monotherapy, patients with compensated liver cirrhosis, and patients with acute infection. It is possible to treat patients with HBV and HIV co-infection, patients with severe extrahepatic manifestations of HCV infection, and patients with transplanted liver. Drug users and alcoholics can be treated after 12-month abstinence. Therapy is not recommended in patients with fulminant hepatitis, patients with persistently normal ALT levels without fibrosis, kidney transplant recipients, and pregnant women. PMID- 16381239 TI - [Treatment of chronic hepatitis B]. AB - More that 350 million people worldwide suffer from chronic hepatitis B virus (HBV) infection. Up to 40 percent of persons infected with the virus may develop complications related to cirrhosis or hepatocellular carcinoma. This reservoire is further growing up despite the fact that more than 110 countries have introduced large-scale vaccination against hepatitis B as part of the worldwide campaign launched by the WHO in 1991 and despite improved blood supply testing. At present, in Croatia there are no precise data on the rate of chronic HBV carriers, however, it is estimated to be < 2% (low prevalence country). To date, two drugs, interferon alfa and lamivudine, have been approved for the treatment of chronic hepatitis B in Croatia. Recent guidelines for the treatment of chronic hepatitis B based on the accumulated knowledge on the disesase pathogenesis and treatment outcome are presented. PMID- 16381240 TI - [Prophylaxis and therapy of hepatitis B after liver transplantation]. AB - About 25% of all liver transplantations in USA are performed for liver disease due to viral hepatitis, 25% of them related to hepatitis B. It is also known that 90%-100% of transplanted persons with proven replicative stage of hepatitis B [HBeAg (+), HBV DNA (+)] develop reinfection of the transplanted liver in the absence of prophylaxis. This percentage is lower (30%-50%) in patients with nonreplicative stage of infection [HBeAg (-), HBV DNA (-)]. The consequence of most transplant infections is death due to the transplanted organ failure. The prophylaxis of infection is possible, however a combination of intravenous hepatitis B hyperimmune gammaglobulin and lamivudin may be too expensive for some transplantation centers in less developed countries. The aim of this article is to recommend prophylaxis and treatment of recurrent hepatitis B in persons with transplanted liver using data of the Consensus Conference on Hepatitis B in Geneva 2002, Canadian Consensus Group for Hepatitis (CASL) and EASL International Consensus Conference on Hepatitis B that should be available for patients in Croatia. PMID- 16381241 TI - [Treatment of recurrent HCV infection after liver transplantation]. AB - End-stage liver disease associated with hepatitis C virus (HCV) infection is the most common indication for orthotopic liver transplantation (OLT) and accounts for 50% of these procedures in Spain and 42% of OLT performed in the United States. Recurrent infection with HCV after OLT, however, is almost universal and is associated with substantial morbidity, mortality, and graft loss. In contrast to immunocompetent individuals, HCV infection in immunosuppressed transplant recipients usually has an accelerated course. Acute hepatitis develops in approximately 75% of HCV recipients in the first six months following OLT. By the fifth postoperative year, over 80% of HCV-infected liver transplant recipients will develop histologic evidence of chronic allograft injury secondary to hepatitis C, with up to 30% developing cirrhosis. While the choice of calcineurin inhibitor has not been clearly shown to affect the histologic recurrence of hepatitis C or the frequency of rejection in HCV-infected recipients, cumulative exposure to corticosteroids is associated with increased mortality, higher levels of HCV viremia and more severe histologic recurrence. There have been no well controlled, large, prospective, multicenter and randomized clinical trials to determine the optimal approach to the treatment of recurrent HCV infection following OLT. Most published studies were small, lacked controls, had short follow-up periods, and were devoid of histologic analysis. Furthermore, most of the published studies are largely incomparable due to differences in the definition of recurrent hepatitis C, timing of anti-HCV therapy administration relative to transplantation, the drugs and doses used and regimens employed, and the study end points assessed (i. e. biochemical, virologic and histologic end points have not all been consistently investigated). In lieu of large studies in post-transplant patients, monotherapy with conventional interferon or monotherapy with pegylated interferon should be considered in recipients with histologically confirmed recurrence of HCV infection. The role of hepatitis C immunoglobulin and new imunosuppression agents in the management of post-transplant HCV infection is still evolving. PMID- 16381242 TI - [Retreatment of patients with chronic hepatitis C]. AB - Impressive gains have been made in the treatment of chronic hepatitis C virus (HCV) infection during the past decade. As the treatment for chronic hepatic C has improved, the question has arisen as to whether patients who failed previous HCV treatment regimens should be retreated. Several recent studies involving retreatment have established that a small but significant increase in sustained virologic response (SVR) resulted when nonresponders to prior interferon monotherapy were retreated with interferon and ribavirin. Given the superior results observed with pegylated interferon and ribavirin in the treatment of naive population, it is now appropriate to consider whether retreating previous relapsers and nonreponders, particularly nonresponders to conventional interferon and ribavirin, will be effective. Such treatment would be most advantageous for patients with advanced fibrosis or cirrhosis who are at the highest risk of developing complications of advanced liver disease, including hepatocellular carcinoma. Namely, there is no doubt that interferon-based regimens may reduce hepatic inflammation in the abscence of SVR. For this reason, patients with advanced liver disease receive histologic benefit from therapy in addition to the potential benefit of viral eradication, because progression of fibrosis to cirrhosis is a function of hepatic inflammation. A number of investigator initiated clinical trials have been started to study response in prior nonresponders and relapsers retreated with pegylated interferon and ribavirin. From the data obtained from these trials, so far is clear that combined therapy can clear HCV-RNA in a significant number of prior nonresponders (25% to 40%) while on treatment, yet the rate of SVR appears to be by far lower (20%). Better results have been reported with relapsers to prior combination therapy. Moreover, preliminary results from the HALT-C trial have shown that maintenance therapy with pegylated interferon and ribavirin can significantly delay fibrosis progression and reduce the risk of hepatic decompensation and hepatocellular carcinoma and the need of liver transplantation. According to these results, many authors have proposed a course of combination therapy with pegylated interferon and ribavirin for relapsers and nonresponders with advanced fibrosis and cirrhosis. PMID- 16381243 TI - [Hepatitis C--treatment of untreated (naive) patients]. AB - The most important therapeutic advance in recent years considering chronic HCV infection has occurred with the introduction of pegylated interferon (PEG IFN) in the combination therapy with ribavirin, which results in better sustained virologic response (SVR). Although an SVR is difficult to correlate with improved survival because of the necessity for long-term follow up, the absence of detectable serum HCV RNA has been associated with resolution of liver injury, reduction in hepatic fibrosis, and a low likelihood of a relapse of the HCV infection. Two large pivotal trials examined the efficacy of PEG IFN plus ribavirin in the treatment of chronic HCV infection. Overall, PEG IFN plus ribavirin was more effective than the standard interferon-ribavirin combination. SVR rates were similar with both forms of PEG IFN (PEG IFN alfa-2a and PEG IFN alfa-2b) when used in combination with ribavirin. SVR rates of 42% and 46% were achieved in patients with genotype 1 compared to rates of 76% and 82% in patients with genotypes 2 and 3. Factors associated with successful therapy included genotypes other than 1, lower baseline viral levels, less fibrosis or inflammation on liver biopsy, and lower body weight or body surface area. Twenty four weeks of treatment with a combination of PEG IFN and ribavirin appears to be sufficient for patients infected with genotypes 2 and 3, while patients with genotype 1 need 48 weeks of treatment. Early virologic response (EVR), defined as undetectable HCV RNA or a minimum 2 log decrease in viral load (relative to baseline) after the first 12 weeks of treatment, is predictive of SVR and should be a routine part of monitoring patients with genotype 1. Patients who fail to achieve an EVR have only a small chance of achieving an SVR, therefore treatment should be discontinued after 12 weeks. It is also recommended to treat patients with acute hepatitis C to reduce the risk of developing chronic infection. Treatment should start 12 weeks after the onset of symptoms and includes 24 weeks of monotherapy with PEG IFN. PMID- 16381244 TI - [Treatment of viral hepatitis in children]. AB - The goal of treating children with chronic hepatitis B is the prevention of complications rather than the urgency of stopping the disease. Candidates for treatment are HBV DNA and HBeAg positive children, older than 2 years, with elevated transaminases for at least 6 months and histologic evidence for chronic hepatitis. Treatment can be carried out with interferon alfa or lamivudine, however, interferon is preferred for higher efficacy. The combined therapy has been currently studied. No consensus has yet been attained on the necessity to treat chronic hepatitis C in children. Candidates for treatment are HCV RNA positive children, older than 3 years, having persistently elevated transaminases, with histologic evidence for chronic hepatitis and significant fibrosis. Until recently, treatment was carried out with interferon alfa, but now, due to better results, a combined therapy with interferon alfa and ribavirin is recommended. New controlled trials are expected as a basis for general recommendations. PMID- 16381245 TI - [Treatment of the side effects of antiviral therapy]. AB - The adverse effects of antiviral drugs are dose dependent and often reversible. The major side effects include influenza-like symptoms, hematologic abnormalities and neuropsychiatric symptoms. The influenza-like syndrome can be prevented by paracetamol taken at the time of the injection. Psychiatric adverse effects range from irritability to a severe depressive syndrome. Antidepressants, such as selective serotonin reuptake inhibitors, may be useful in the management. Adverse hematologic effects can occur very early during treatment. The platelet count often stabilizes rapidly, but neutropenia can deteriorate throughout treatment. In selected patients treatment with hematopoietic growth factor (filgrastim) may be useful. Ribavirin therapy may result in a dose-dependent reversible intravascular hemolytic anemia in 10% of patients. Adjunctive therapy with erythropoietin for ribavirin-induced anemia is currently under evaluation. Interferons and ribavirin are contraindicated in pregnancy. Contraception must be continued for 4 months (women) and 7 months (men) after ribavirin cessation. Lactic acidosis may be a rare complication of combination therapy in patients undergoing therapy for HIV and HCV. Any sign of mitochondrial DNA depletion syndrome calls for blood lactate measurement and, possibly, a modification of antiretroviral treatment. Lamivudine is well tolerated but the emergence of lamivudine-resistant (YMDD) HBV mutants is associated with the loss of clinical response. Adefovir dipivoxil effectively suppresses lamivudine-resistant HBV in chronic hepatitis B. PMID- 16381246 TI - [Treatment of chronic hepatitis C in patients with HIV infection]. AB - Treatment of chronic hepatitis C in patients infected with the human immunodeficiency virus (HIV) is recommended when: 1) aminotransferase levels are repeatedly elevated; 2) CD4+ blood cell counts are above 350 per microl; and 3) HIV RNA plasma levels are less than 50 000 copies per milliliter. Treatment is not recommended for patients who inject illegal drugs, consume large amount of alcohol, or have a severe psychiatric disorder. Treatment of patients with normal aminotransferase levels can be considered in the context of a clinical trial or if stage F2 or worse has been histologically confirmed on a liver biopsy specimen. Liver biopsy is generally recommended prior to treatment. However, because of faster progression to fibrosis in HIV and hepatitis C virus (HCV) coinfected patients, if the patient declines liver biopsy it should not exclude him from treatment. Treatment with interferon and ribavirin (800 mg/day orally) is recommended. Pegylated interferon is preferred (180 microg of alfa-2a form and 1.5 mg/kg of alfa-2b form once weekly subcutaneously) because of poor results with the conventional form of interferon, however, treatment with conventional interferon (3 times weekly 3 million units subcutaneously) can be considered in HCV genotype 2 or 3 infection. All genotypes should be treated for 48 weeks. Since only patients who have a decline of HCV viremia of at least 2 logarithms after 12 weeks of treatment have a chance of cure, treatment should be discontinued in patients who do not achieve this target. Concurrent treatment with zidovudine and didanosine should be avoided because of additive toxicity. One should also be cautious when antiretroviral drugs with a greater potential for hepatotoxicity (nevirapine, ritonavir) are concurrently administered. PMID- 16381247 TI - [Prevention and treatment of hepatitis B in patients on hemodialysis and vaccination of hemodialysis health personnel against hepatitis B]. AB - The prevalence and incidence of hepatitis B in hemodialysis patients in Croatia have been estimated to 1.3% and 0.03%, respectively. HBV infection in dialysis patients is usually asymptomatic, has a prolonged course, and progresses to chronic HBsAg hepatitis in 50% of cases. Some 15%-40% of HBsAg carriers on dialysis will develop cirrhosis, liver decompensation or hepatocellular carcinoma. Strict adherence to the standard infection prevention measures, continuous monitoring of HBV markers in patients on hemodialysis, patient and personnel immunization and hepatitis B treatment in hemodialyzed patients are mandatory. Each new patient in a dialysis center must be tested for HBV markers irrespective of prior immunization. All patients in the center should be routinely screened every 3-4 months. HBV immunization is mandatory for all patients on dialysis. In patients with uremia the anti-HBs antibody production is decreased (antibodies will develop in 50%-60% of cases after immunization). It is recommended to immunize all patients with progressive kidney disease, preferably in the preterminal stage. Hepatitis B therapy is recommended in all patients with biopsy proven chronic liver disease. Patients should be treated with standard interferon alpha and/or lamivudine, or peginterferon alpha monotherapy. Hepatitis B treatment is most important in kidney and/or liver transplant candidates. HBV immunization is obligatory for all hospital personnel who are in close contact with infected patients and infective materials. PMID- 16381248 TI - [Prevention and treatment of hepatitis C in patients on dialysis]. AB - Nosocomial transmission of blood-borne pathogens, including hepatitis C virus infection, is the most common one in a dialysis center setting. The prevalence of HCV antibodies is by far higher in patients undergoing maintenance hemodialysis therapy than in those on peritoneal dialysis. Standard infection prevention measures in hospital settings and measures of infection prevention in dialysis units should be performed. They include serologic testing for HCV of every new patient in a dialysis unit as well as routine testing of all patients every six months. Hepatitis C therapy is recommended in patients on dialysis who have detectable HCV RNA, positive liver biopsy (portal or bridging fibrosis or moderate stage of necroinflammation), younger patients (less than 65 years), and transplantation candidates. When evaluating ALT, it should be kept in mind that ESRD patients have ALT levels lower than general population, making ALT level not relevant parameter of liver disease activity in these patients. Results of hepatitis C therapy with interferon alpha and peginterferon alpha are similar to those in the general population but with more common side effects, which may require therapy discontinuation. Due to the possibility of anemia, ribavirin is contraindicated in patients with ESRD. Around 30% of patients treated with peginterferon have sustained viral response, 25%-45% of them have end of treatment viral response, and 50%-80% have end of treatment biochemical response (ALT normalization). Numerous clinical trials have established that the decrease in HCV load and prolonged suppression of viral replication during interferon therapy significantly reduce hepatic inflammation and consequently postpone progression of fibrosis to cirrhosis. PMID- 16381249 TI - Loss of licensure as a result of the conviction of a crime. PMID- 16381250 TI - A look at POs and PHOs in Michigan. PMID- 16381252 TI - That is what I do. PMID- 16381251 TI - Pay-for-performance: whether fad or foundation, doctors must be involved now. PMID- 16381253 TI - Effective risk management pays off. PMID- 16381254 TI - The perfect storm, Part II. PMID- 16381255 TI - Drug interactions in anesthesia. PMID- 16381256 TI - Does pancuronium or cisatracurium delay the rate of arousal following remifentanil-based anesthesia? AB - STUDY OBJECTIVE: The present report investigates the rate of arousal following remifentanil-based anesthesia associated with the coadministration of pancuronium, which inhibits butyrylcholinesterase, or cisatracurium, which is partially metabolized by nonspecific esterases, versus vecuronium that is eliminated independently of ester hydrolysis. DESIGN, SETTING AND PATIENTS: Sixty patients, ASA I-II, scheduled for elective abdominal surgeries were enrolled in a double-blinded prospective study. In fact, patients were equally divided into three Groups with each Group receiving remifentanil and either one of the following three muscle relaxants: pancuronium, vecuronium or cisatracurium. MEASUREMENTS: The rate of arousal following discontinuation of anesthesia was assessed by Modified Aldrete Score. Time to eye opening on verbal command, tracheal extubation, Modified Aldrete Score >9, and time to discharge from the recovery room were recorded. MAIN RESULTS: Time to eye opening on verbal command, tracheal extubation, Modified Aldrete Score >9, and time to discharge from the recovery room were not significantly different between the three groups. CONCLUSION: The results suggest that recovery following remifentanil-based anesthesia is not delayed by the coadministration of pancuronium, cisatracurium versus vecuronium; and by the use of neostigmine for reversal of neuromuscular blockade. PMID- 16381257 TI - Cricoid pressure: an evidence-based practice? PMID- 16381258 TI - Anesthesia and care for the patient with diabetes mellitus and associated complications. PMID- 16381259 TI - Induction-intubation response--smokers vs non-smokers--. AB - BACKGROUND AND OBJECTIVES: Tobacco smoking has been established to be a hazardous activity. Changing social attitude is bringing a decline in tobacco consumption but a significant proportion of patients presenting for surgery still continues to smoke, putting themselves at risk of perioperative complications. We evaluated induction-intubation response in 40 male patients (ASA-I) divided into two groups of 20, each consisting of smokers and non-smokers. METHODS: All patients received standard premedication and general anesthesia. Heart rate, blood pressure and rate-pressure product were measured prior to induction, just before intubation and at 1, 3, 5 and 10 minutes after intubation. Continuous electrocardiography (EKG lead II) monitoring was done. Carboxyhemoglobin and total hemoglobin were estimated at the time of securing venous access. Arterial blood gases were analysed 5 minutes before and after the tracheal intubation. RESULTS: Incidence of arrhythymias (30%) was higher in smokers compared to non-smokers (10%). Mean carboxyhemoglobin (COHb) level in smokers was 3.81 +/- 2.17 gdL(-1) as compared to 2.95 +/- 1.33 gdL(-1) in non-smokers. Four patients who continued to smoke till the day of surgery had higher COHb levels (8.2, 5.9, 6, 8.8 gdL(-1)). PaO2 and PaCO2 levels were comparable in both groups. CONCLUSIONS: During induction intubation period, heart rate; systolic, diastolic and mean arterial pressure and rate-pressure product showed more pronounced fluctuations in smokers than in non smokers (p < 0.05). PMID- 16381260 TI - The challenge of a multicultural environment for critical care nursing in Saudi Arabia. AB - With a worldwide shortage of critical care nurses and the alluring opportunity for nurses to work abroad, it can be said that the multicultural experience associated with working in Saudi Arabia may not be unique to the Kingdom. Healthcare staff is faced with many challenges in a multicultural environment in relation to language, customs, communication, and healthcare practices. In this article it is endeavored to discuss these challenges and to suggest some possible solutions to the problems encountered. PMID- 16381261 TI - The intubating laryngeal mask airway Fastrach for emergence after carotid endarterectomy. AB - We determine the feasibility of using the intubating laryngeal mask airway Fastrach (ILM) as a ventilatory device during emergence from anesthesia after use as an airway intubator in patients undergoing carotid endarterectomy. Thirty-five patients (ASA 2-3, 53-84 yr) were studied. Induction was with midazolam/fentanyl/etomidate and maintenance was with sevoflurane 1-2% in O2 33 50% and N2O. Neuromuscular blockade was with cisatracurium. Tracheal intubation was with a flexible lightwand via the ILM. After successful intubation, the ILM remained in the pharynx, but with the cuff deflated. After surgery, but before anesthesia was discontinued, baseline cardiovascular variables were recorded. The ILM cuff was then reinflated, the tracheal tube removed, the anesthesia breathing system connected to the ILM and anesthesia discontinued. Any changes in the cardiovascular variables greater than +/- 20% baseline values were noted from cuff reinflation to 1 minute after ILM removal. Any adverse respiratory (laryngospasm, coughing, gagging, stridor, SpO2 <94%, end-tidal carbon dioxide >45 mmHg, regurgitation/aspiration) or electrocardiographic (ST segment or rhythm changes) events were also noted. Patients were questioned about postoperative sore throat at 2 and 24 hr. ILM insertion and intubation through the ILM were successful in all patients. Adequate ventilation was achieved in all patients before intubation and after extubation. The mean (range) time taken from cuff reinflation to ILM removal was 9 (5-21) min. The rate pressure product remained within +/- 20% baseline values in all patients. There were no adverse respiratory or electrocardiographic events. There were no adverse neurological events. The surgical field was satisfactory. Postoperative sore throat occurred in 14% at 2 hr and 0% at 24 hr. We conclude that the ILM can be used as a ventilatory device for emergence from anesthesia after use as an airway intubator for carotid endarterectomy. PMID- 16381262 TI - The patient with maple syrup urine disease. PMID- 16381263 TI - Perioperative bioelectrical impedence analysis in neurosurgery. AB - The use of bioelectrical impedence (BI) measurement to assess body composition has recently attracted the attention of anesthesiologists. Analysis of BI provides a non-invasive method to quantify fluid distribution in different body compartments. This study was designed to assess whether BI analysis reflects fluid depletion in neurosurgical patients with moderate blood loss. Six adult male patients scheduled for elective craniotomy under general anesthesia were studied. Exclusion criteria included patients with cardio-respiratory disease. BI analysis was performed at three stages, A, day before operation, B, during surgery and C, on the first postoperative day. Total body resistivity was measured by BI analysis with a four-terminal portable impedence analyzer. At each frequency, impedence was calculated as resistance (Rx)2 + reactance (Rc)2. The mean values of total body water (TBW) at stages A, B and C were 39.8 L (range: 33.1-46.7 L), 43.2 L (range: 33.1-66.2 L) and 36.8 L (range: 22.4-36.3 L) respectively with significant differences (P<0.05). The impedence at the three frequencies during stages A, B and C showed significant differences (P<0.05). In conclusion, we have found that in male neurosurgical patients multiple frequency BI measurements has reflected fluid balance perioperatively. Whether this observation remains true for other surgical procedures with massive blood loss, yet to be further investigated. PMID- 16381264 TI - Remifentanil versus propofol sedation for peribulbar anesthesia. AB - The short onset and offset of remifentanil may allow for accurate dosing of sedative effect with few side-effects and rapid recovery. In this study, remifentanil is compared with propofol for sedation intraocular pressure hemodynamics and oxygen saturation, in patients undergoing cataract surgery with peribulbar block. After ethical committee approval, fifty patients scheduled for one eye cataract surgery were enrolled into the prospective study. Remifentanil infusion was initialized at a rate of 0.04 microg/kg/min, 10 min before peribulbar anesthesia in Remifentanil group (Group R). In Propofol Group (Group P), propofol infusion of 2 mg/kg/h was reduced to 1mg/kg/h. Intraocular pressures of the contralateral eye were measured before and after peribulbar anesthesia and at the end of the operation. Sedation scores (1-5), hemodynamic parameters and saturation were assessed at 5 minute intervals. Heart rate, intraocular pressure, saturation and sedation scores were similar between groups. Intraocular pressures decreased within time in both groups. There were significant changes in mean blood pressure after 5th min between groups (P < 0.05). The infusion of remifentanil or propofol for sedation provided adequate sedation, hemodynamic stability without increasing intraocular pressure during surgery in patients undergoing cataract surgery with peribulbar anesthesia. PMID- 16381265 TI - Wound instillation with 0.25% bupivacaine as continuous infusion following hysterectomy. AB - Postoperative pain relief was assessed by the effects of local anesthetic wound instillation on 100 patients who had undergone total abdominal hysterectomy with bilateral salpingo oophorectomy (TAH with BSO). Patients were divided into four groups of wound and non-wound instillation: Wound instillation Group A1 received diclofenac IM. Group A2 received diclofenac suppository. Non-wound instillation Group B1 received diclofenac IM. Group B2 received diclofenac suppository. A standard general anesthesia technique was administered. For would instillation, a multiholed (1 cm apart) 18G epidural catheter was placed above rectus sheath. This was connected to a pediatric regulated drip set with "Dial-a flo" to deliver 0.25% bupivacaine 10 ml/hour for 6 hours after a basal bolus of 10 ml. During first 6 hours after surgery rescue pentazocine 15 mg was administered to achieve VAS score < or = 30. Thereafter, rescue diclofenac was administered to patients. The requirement of rescue analgesic (pentazocine) was significantly less (P < 0.001) in wound instillation Groups A1 (13.80 mg +/- 13.64) and A2 (12.00 mg +/- 12.25) in comparison to non wound instillation Groups B1 (35.60 mg +/- 14.02) and B2 (31.80 mg +/- 15.80). Rescue diclofenac was not required in wound instillation groups as compared to 30 mg (B1) and 36 mg (B2) in non wound instillation groups. Nausea and vomiting was less in wound instillation groups. VAS score supine from 4th to 12th hours, VAS coughing during all time interval and VAS leg raising from 3rd to 12th hours was significantly lower (P < 0.001) in wound instillation group (A1, A2) in comparison to non wound instillation groups (B1, B2). We conclude that basal bolus infusion followed by continuous wound instillation of bupivacaine decreases analgesic requirement and pain scores in first 24 hours of postoperative period after TAH with BSO. PMID- 16381266 TI - Epidural for labour analgesia--bupivacaine + fentanyl vs bupivacaine + fentanyl + epinephrine. AB - A prospective and randomized study was conducted on 50 full term parturients undergoing labour analgesia at Panna Dai Hospital, R.N.T. Medical College, Udaipur. The parturients were allocated in two groups: group BF (n = 25) received an epidural injection of bupivacaine (0.15%; 15 mg) + Fentanyl (0.0002%); 2 microg/ml); group BEF (n = 25) bupivacaine (0.15%; 15 mg) + Fentanyl (0.0002%; 2microg/ml) + Epinephrine (1.7 microg/ml; 1:600,000). Subsequent top up (same as bolus) was administered at VAS > or = 3. Comparison between the two groups showed no significant difference in onset of analgesia, number of top up doses, degree of motor block and parturients acceptance. The difference in duration of analgesia with addition of epinephrine (93.00 +/- 29.00 min : BEF group; V/S 86.00 +/- 27.00 Min : BF group) did not show any statistical significance between the two groups. Ambulation was achieved in 100% of parturients from both study groups. The incidence of spontaneous delivery was 92% in each group. Parturients in group BEF demonstrated a higher incidence of side effects like nausea and vomiting (4% v/s 0%); hypotension (4% v/s 0%); paresis (8% v/s 0%) and retention of urine (12% v/s 8%) as compared to BF group. To conclude, the addition of epidural epinephrine (1:600,000) to bupivacaine and fentanyl does not affect the duration or quantity of labour analgesia. PMID- 16381267 TI - The effect of prone position on respiratory mechanics during spinal surgery. AB - OBJECTIVE: To study the effect of prone position on respiratory mechanics during spine surgery. DESIGN: Prospective study. SETTING: Elective spine surgery at a university hospital. PATIENTS: 12 ASA physical I & II with no coexisting cardiorespiratory disease undergoing cervical or lumbar laminectomy under general anesthesia in prone position. MEASUREMENTS: Ten min after induction of general anesthesia and endotracheal intubation, while patients were in supine position, the following measurements were taken using anesthesia delivery unit (Datex Ohmeda type A_Elec, Promma, Sweden): peak airway pressure (Ppeak), peak plataeu pressure (Pplat), peak mean pressure (Pmean) and dynamic lung compliance (DLC). The same measurements were recorded 10 min after placing patients into prone position. At the end of surgery and 5 min after turning the patients supine and before tracheal extubation, the same measurements were again recorded. The results expressed as means +/- sd. One way ANOVA was used for analysis of differences in the data before, during prone position and after turning patients supine at the end of the procedure. For all comparisons p < 0.05 was considered significant. RESULTS: During prone position there was significant reduction in DLC and significant increase in airway pressures. CONCLUSION: We conclude that turning the patients form supine to prone position during anesthesia for spine surgery caused significant decrease of DLC and significant increase of airway pressure. PMID- 16381268 TI - Bilateral negative airway pressure pulmonary edema (NPPE)--a case report--. AB - A case of negative pressure bilateral pulmonary edema in a 28 years old healthy female patient, scheduled for diagnostic pelvic laparoscopy for infertility. Following extubation and apparent recovery from anesthesia, she had strong inspiratory efforts due to airway obstruction caused by coughing and laryngeal spasm, that lead to negative pressure bilateral pulmonary edema. The pulmonary edema disappeared within few hours. She was breathing spontaneously through CPAP system (mask-bag-expiratory valve). Diuretics and lungs physiotherapy helped in controlling patient's complication. PMID- 16381269 TI - Anesthetic management of autoimmune polyglandular syndrome (Schmidt's syndrome)- a case report-. AB - BACKGROUND: Autoimmune polyglandular syndromes (APS) are complex diseases with diverse clinical presentations resulting from involvement of multiple endocrine glands. Surgery under anesthetic in these patients is challenging. A case of Schmidt syndrome (autoimmune polyglandular syndrome type 11) that developed adrenocortical insufficiency in the postoperative period is reported. Etiology, pathogenesis, types and anesthetic problems associated with these cases are discussed. CASE REPORT: A 41 yr old female patient, diagnosed to have APS (Schmidt syndrome) presented for uterine surgery. She had autoimmune glandular involvement of pituitary, thyroid, parathyroid, adrenals and melanocytes and was on hormone replacements for the deficiencies incurred, which were continued till the morning of surgery. Surgery was conducted under general anesthetic combined with epidural analgesia. In spite of supplementation of steroid in physiological doses prior to surgery, she developed hemodynamic instability in the early postoperative period, but could be successfully resuscitated with additional steroid dosage and fluids. CONCLUSION: This patient presented with multiglandular endocrine involvement necessitating timely, adequate hormone replacement and appropriate fluid management. These challenges require careful approach to anesthetic management. PMID- 16381270 TI - Anesthetic problems in Christ Siemens Touraine syndrome--a case report--. PMID- 16381271 TI - An audit of postoperative minor complications of anesthesia. AB - OBJECTIVE: To look at minor complications attributable to anesthesia in adult surgical patients at our Institution and to identify various contributing factors. DESIGN: Descriptive cross sectional audit. SETTING: Aga Khan University Hospital, Karachi, Pakistan. STUDY PARTICIPANTS: Seven hundred and thirty surgical patients undergoing elective surgery were recruited. INTERVENTIONS: This was a non-interventional study and data was collected prospectively based on predefined criteria. MAIN OUTCOME MEASURES: The incidence of nausea, vomiting, sore throat, headache, drowsiness, phlebitis, dizziness, myalgia, transient nerve palsy, conjunctivitis, ringing of ears, low backache, lip injury, dental injury or any other minor complications were looked at. RESULTS: Nausea, sore throat and vomiting were the highest reported complications. The incidence of nausea, drowsiness and dizziness was less in patients more than sixty years of age. Incidence of nausea and vomiting was higher in ASA 1 patients and in surgery lasting less than ninety minutes. Headache and dizziness were reported high in certain surgical specialties. CONCLUSION: The overall rate of minor complications following anesthesia was 12.6%. No complications were reported by 30% of the study population. The data has given us a benchmark for patient information and will be used for risk reduction in our Department of Anaesthesiology. PMID- 16381272 TI - Spinal fusion surgery obviating bone graft harvest--a reality in the near future. PMID- 16381273 TI - The Malaysian Orthopaedic Association humanitarian mission to Indonesia and Sri Lanka. AB - The tsunami which occurred off the west coast of North Sumatra on December 26, 2004 devastated the coastal areas of North Sumatra, South-West Thailand, South East India and Sri Lanka killing more than a quarter of a million people. The destruction was enormous with many coastal villages destroyed. The other countries affected were Malaysia, Myanmar, Maldives, Bangladesh, Somalia, Kenya, Tanzania and the Seychelles. In January 2005, volunteers went in weekly rotation to Banda Aceh in collaboration with Global Peace Mission. These were Dr Hyzan Yusof, Dr Suryasmi Duski, Dr Sharaf Ibrahim, Dr Saw Aik, Dr Kamariah Nor and Dr Nor Azlin. In Banda Aceh, the surgical procedures that we could do were limited to external fixation of open fractures and debriding infected wounds at the Indonesian Red Crescent field hospital. In February, a team comprising Dato Dr K S Sivananthan, Dr T Kumar and Dr S Vasan spent a week in Sri Lanka. In Sri Lanka, Dato Sivananthan and his team were able to perform elective orthopaedic operations in Dr Poonambalam Memorial Hospital. We appealed for national and international aid and received support from local hospitals and the orthopaedic industry. International aid bound for Banda Aceh arrived in Kuala Lumpur from the Philippine Orthopaedic Association, the Chiba Children's Hospital in Japan and the Chinese Orthopaedic Association. The COA donated 1.5 tons of orthopaedic equipments. A special handing over ceremony from the COA to the Indonesian Orthopaedic Association was held in Putrajaya in March. Malaysia Airlines flew in the donated equipment to Kuala Lumpur while the onward flight to Aceh was provided by the Royal Malaysian Air Force. In April, Dr Saw Aik and Dr Yong Su Mei joined the Tsu-Chi International Medical Association for volunteer services on Batam Island, Indonesia. The MOA acknowledges the many individuals and organizations, both governmental and non-governmental, for their contributions in the humanitarian efforts. PMID- 16381274 TI - Extracorporeal shock wave therapy for musculoskeletal pathology--a literature review. AB - For more than two decades extracorporeal shock wave lithotripsy has emerged as the standard therapy for calculi in the kidney and urinary tract, and biliary system. Application of extracorporeal shock waves in orthopaedics involves treatment of recalcitrant chronic pain of plantar fasciitis, tennis elbow and calcifying tendonitis of the shoulder. This review explores current evidence based issues related to its potential use as a treatment option for some musculoskeletal conditions. PMID- 16381275 TI - Cycle spokes and chain related injuries in children--emerging injuries in Malaysia. AB - A descriptive prospective study of 16 children with injuries inflicted by bicycle spokes and chain was undertaken to identify the demographic profiles of such injuries. Aspects of preventive measures are proposed. PMID- 16381276 TI - The incidence of deep venous thrombosis following arthroscopic knee surgery. AB - A prospective cohort study was undertaken at two centers to look for the incidence of deep vein thrombosis (DVT) following knee arthroscopic surgery. Eighty-four patients who had 90 arthroscopic procedures were reviewed. The relevant risk factors: past or family history of DVT, smoking, oral contraceptives, body weight, haemoglobin level, platelet count, tourniquet time and type of anaesthesia were documented. All patients were subjected to preoperative and post-operative duplex ultrasound. Only one patient (1.2 %) was noted to have DVT involving the peroneal vein. We concluded that the incidence of DVT after knee arthroscopy was very low in this study population. PMID- 16381277 TI - The epidemiology of shoulder dislocation in a state-hospital: a review of 106 cases. AB - This retrospective study was conducted in a state hospital set-up and aimed at identifying the magnitude of shoulder dislocations and their demographic data, characteristics of the injury, mechanism and predisposing factors, and the instituted treatment. Patients with radiographic evidence of shoulder dislocation admitted to the hospital from January 1999 to December 2002 were included. Data were recorded from the case notes. There were 105 shoulder dislocations with male predomination in 77% cases and age ranged between 11 and 90 years (average 30.9 years). The right shoulder was affected in 68% of the cases. The contributing events were fall in 37% of cases, road traffic accident 23%, sports 17% and pathological conditions 13%. Anterior dislocation occurred in 96.2% of the cases. Posterior and inferior dislocations encountered in two patients for each type. Twelve dislocations were associated fracture of the greater tuberosity, two each with humeral neck fracture and cerebral injuries. First time dislocation occurred in 73.6% of the cases. The recurrences ranged between 2 to 6 times (average 3.4 times). Closed manipulative reduction and strapping was the definitive treatment in 92.4% of the cases and the remaining needed surgical reconstruction. Four patients had open reduction and internal fixation of the associated fractures while another four had arthroscopic Bankart's repair. In conclusion, shoulder dislocation represents the most common shoulder problems. It afflicted young adults of reproductive age (21-40 years) and participation in sports was a risk factor in men. Women over 40 years and fall were at risk to develop shoulder dislocation. PMID- 16381278 TI - Sports-related shoulder dislocations: a state-hospital experience. AB - This retrospective study was conducted in a state hospital set-up and aimed at identifying the incidence of sports-related shoulder dislocations and their characteristics and the sports events involved. All patients with shoulder dislocation related to sporting activities admitted to the hospital from January 1999 to December 2002 were included in the study. There were 18 sports-related shoulder dislocations out of 106 all shoulder dislocations admitted during this 4 year period. The average age of the patients was 25.4 years. All but two were male. All were anterior dislocations. Recurrent dislocation constitutes 78% of the cases with an average of 3 times re-dislocation. Rugby and badminton were the major contributors to the injuries followed by volleyball, soccer and swimming. Conservative treatment was successfully instituted for 88% of the patients and 12% opted for surgical intervention. PMID- 16381279 TI - Radiation exposure to the surgeon during femoral interlocking nailing under fluoroscopic imaging. AB - Femoral interlocking nailing requires fluoroscopic assistance for insertion of the nail and distal screws. In this study, scattered radiation to the eye and hand of the operating surgeon was measured during the procedure. Thermo luminescent dosimeter (TLD) was used to quantify the dose received by the surgeon. The mean radiation exposure time during the procedure was 3.89 minutes. The mean scattered radiation doses to the hand and eye were 0.27 mSv and 0.09 mSv per procedure respectively. These very low doses have made a surgeon very unlikely to receive more than the recommended annual dose limit set by the National Council on Radiological Protection. PMID- 16381280 TI - The HUKM Spinal Instrumentation System for adolescent idiopathic scoliosis. A biomechanical comparison study using finite element analysis. AB - This finite element analysis is aimed at comparing relative stiffness of three different posterior instrumentation constructs: the Hospital Universiti Kebangsaan Malaysia Spinal Instrumentation System (HUKM-SIS), the Cotrell Dubousset Instrumentation (CDI) and Harrington Instrumentation System (HIS), used in the treatment of adolescent idiopathic scoliosis (AIS). The constructs were tested under various loads using MSC Patran 2001 r2a. Under increasing flexion loads, there was a linearly corresponding increase in deflection magnitudes for all constructs on the load-deflection curve. The CDI was the stiffest construct under axial, forward flexion and extension loads, followed by the HUKM-SIS and HIS. Under lateral bending loads, the HUKM-SIS construct was the stiffest followed by CDI and HIS. The HUKM-SIS construct was stiffer than HIS under torsional loads. We conclude that multiple pedicle screws increase the stiffness of posterior instrumentation constructs under all loads and inter-segmental spinous processes wiring increase the stiffness against lateral bending. PMID- 16381281 TI - Accuracy of the funnel technique of thoracic pedicle screws insertion in scoliosis surgery--an evaluation by CT-scans. AB - Pedicle screw system has increasingly been used for correction of thoracic scoliosis. It offers several biomechanical advantages over hook system as it controls all three-column of the spine with enhanced stability. Of many techniques of pedicle screw placement in the thoracic spine, the funnel technique has been used in Sarawak General Hospital since 2002. This prospective study aims to assess the accuracy of the placement of thoracic pedicle screws using the funnel technique in the corrective surgery of idiopathic scoliosis. A total of 88 thoracic pedicle screws were inserted into the T4 to T12 vertebrae of 11 patients. Post-operative CT-scan was performed to evaluate the position of the pedicle screw. Seventy six (86.4%) screws were noted to be totally within the pedicle. There was no screw with medial violation of the pedicle, 8 (9.1%) screws breeching the lateral wall of the pedicle and 4 (4.5%) screws with anterior and lateral penetration of the vertebral body. No clinical sequel with the mal positioned screws was noted. In conclusion, the funnel technique enabled simple, accurate and reliable insertion of pedicle screw even in the scoliotic thoracic spine without the need of any imaging guidance. It is however imperative for the surgeon to have a thorough knowledge of the thoracic spine anatomy, and to be familiar with the technique to insert these screws diligently. PMID- 16381282 TI - Measurement of patellar thickness in relation to patellar resurfacing. AB - Patellar thickness is an important consideration for resurfacing in total knee arthroplasty. A patella of 25 mm in thickness is not suitable for resurfacing using the currently available total knee systems. A cross sectional study on patellar thickness using plain radiographs was undertaken on 56 patients. It was observed that the average thickness of the bony part of the patella was 20.05 mm (range 17-23 mm). The actual thickness of cartilage varies from 2.0 to 5.5 mm (mean 3.2 mm). Thus the mean thickness of the patella was 23.2 mm (range 20.2 26.2 mm). The majority of our patients (73%) had patellar thickness of 24 mm or less. The patellar thickness had a significant correlation with the patient's height (R=0.5). Since the majority of our patients have thin patella, we are of the opinion that majority of our patients are not suitable for patellar resurfacing. PMID- 16381283 TI - Modular endoprosthetic replacement after total femur resection for malignant bone tumor. AB - Total femur endoprothesis is an alternative replacement for massive malignant bone tumor with intramedullary extension or skip lesion. Four patients underwent total femoral resection and replacement with megaprosthesis: three had primary malignant bone tumor and one had salvage procedure for aseptic loosening of the distal femoral replacement. Tumor-free margins were achieved in all patients with two patients required vascularized latissimus dorsi free flap cover for reconstruction of soft tissue defects. The average follow-up was 24 months (range 16 - 60 months). All four patients were still alive with three of them being disease-free and one survived even with the presence of lung metastasis. The functional results obtained were either excellent or good in all patients in accordance to the Musculoskeletal Tumors Society grading system. PMID- 16381284 TI - Colonies in engineered articular cartilage express superior differentiation. AB - In view of poor regeneration potential of the articular cartilage, in-vitro engineering of cartilage tissue offers a promising option for progressive joint disease. This study aims to develop a biologically engineered articular cartilage for autologous transplantation. The initial work involved determination of chondrocyte yield and viability, and morphological analysis. Cartilage was harvested from the knee, hip and shoulder joints of adult New Zealand white rabbits and chondrocytes were isolated by enzymatic digestion of the extra cellular matrix before serial cultivation in DMEM/Ham's F12 media as monolayer cultures. No differences were noted in cell yield. Although chondrocytes viability was optimal (>93%) following harvest from native cartilage, their viability tended to be lowered on passaging. Chondrocytes aggregated in isogenous colonies comprising ovoid cells with intimate intracellular contacts and readily exhibited Safranin-O positive matrix; features typically associated with articular cartilage in-vivo. However, chondrocytes also existed concurrently in scattered bipolar/multipolar forms lacking Safranin-O expression. Therefore, early data demonstrated successful serial culture of adult chondrocytes with differentiated morphology seen in established chondrocyte colonies synthesizing matrix proteoglycans. PMID- 16381285 TI - Bone graft substitute using hydroxyapatite scaffold seeded with tissue engineered autologous osteoprogenitor cells in spinal fusion: early result in a sheep model. AB - Spinal fusion using autologous bone graft is performed in an increasing rate for many spinal disorders. However, graft harvesting procedure is associated with prolonged operation time and potential donor site morbidity. We produced an engineered 'bone graft' substitute by using porous hydroxyapatite (HA) scaffold seeded with autologous bone marrow osteoprogenitor cells (OPCs) and fibrin. This obviates bone graft harvesting, thus eliminates donor site morbidity and shortens the operation time. The aim of this study is to evaluate Hydroxyapatite (HA) ceramics as scaffold for autologous tissue engineered bone construct for spinal fusion in a sheep model. The sheep's marrow was aspirated from iliac crest. The bone marrow mesenchymal stem cells (BMMSCs) were cultured for several passages in the presence of growth and differentiation factors to increase the number of OPCs. After the cultures reached confluence, they were trypsinized and seeded on Hydroxyapatite scaffold (HA). Approximately 5 million cells were generated after 3 weeks of culture. Microscopically, very tight Colony Forming Units (CFU-Fs) were seen on monolayer culture. The Von Kossa and Alizarin Red staining of monolayer culture showed positive mineralization areas; indicating the presence of OPCs. Sheep underwent a posterolateral spinal fusion in which scaffolds with or without OPCs seeded were implanted on both sides of the lumbar spine (L1-L2). Intended fusion segments were immobilized using wires. At the end of third month, the fusion constructs were harvested for histological examination. Fibrous tissue infiltration found in the inter-connecting pores of plain HA ceramics indicates inefficient new bone regeneration. New bone was found surrounding the HA ceramics seeded with autologous cells. The new bone is probably formed by the sheep BMMSCs that were initially encapsulating HA while it remained intact. The new bone is naturally fused with the vertebrae. In conclusion, the incorporation of autologous bone marrow cells improved the effectiveness of HA ceramics as 'bone graft' substitute for spinal fusion. PMID- 16381286 TI - Dynamic traction and passive mobilization for the rehabilitation of zone II flexor tendon injuries: a modified regime. AB - This retrospective study aims to evaluate the effectiveness of the modified regime for rehabilitation of Zone II flexor tendon injuries in Sibu Hospital. From January to December 2003, 8 patients with 15 injured digits were treated by using the combined method of dynamic traction and passive mobilization. According to Strickland's criteria, 14 (93.3%) digits achieved good to excellent outcomes and only 1 (6.7%) was rated as poor. No occurrence of tendon rupture was noted. The overall grip strength of the injured hand was 50.1% of the uninjured hand at 3 months after the repair. Our results compare favorably with the other published studies. We believed that this modified regime is as effective as other established regimes and suitable to be adopted in our setting. Further study with larger sample group will be required to consolidate our findings. PMID- 16381287 TI - Resection arthrodesis for primary bone tumour about the knee. AB - Bone defect following en bloc resection of primary bone tumor around the knee can be reconstructed by allograft or prothesis or combination of both. Resection arthrodesis is an alternative option for young vigorous patients facing circumstances of financial constrain or limited supply of allograft. This study was undertaken to determine the outcome and complications associated with resection-arthrodesis of 22 primary bone tumors (13 giant cell tumors and 9 osteosarcomas) around the knee treated between 1990 and 2003 at the University Malaya Medical Center. The mean follow-up was 6 years (range 1-13 years). hree patients with osteosarcoma died of lung metastasis, 3 required above knee amputation and 2 defaulted follow-up. Local complications of the procedure include infection in 8 cases (36.4%), non-union 7 (31.8%) and mal-union. Of 14 patients who returned for final evaluation, 79.8% had satisfactory outcomes according to the Musculoskeletal Tumor Society grading system. In conclusion, resection-arthrodesis of the knee is a viable treatment option for selected patients with primary bone tumor around the knee, and good functional outcome can be expected in the presence of short-term local complications. PMID- 16381288 TI - Shoulder arthrodesis in brachial plexus injuries--a review of six cases. AB - Shoulder arthrodesis is a secondary reconstructive option for patients with brachial plexus injuries requiring a stable shoulder. This study was undertaken to evaluate the clinical and radiological outcomes of shoulder fusion in six patients with flail upper limbs following complete brachial plexus injuries. The shoulder was fused in 30 degrees abduction, 30 degrees internal rotation and 30 degrees flexion via a direct lateral approach by using a 4.5 mm reconstruction plate without bone grafting. The average follow-up was 10.3 months. Radiological union was obtained in all patients. Five patients (83%) had relief of pain after the shoulder fusion. Improvement of function was observed in all patients with a mean improvement of 56.6 degrees (range 30 degrees-75 degrees) and 47.5 degrees (range 30 degrees-60 degrees) active flexion and abduction respectively. One patient developed humeral fracture distal to the plate and the fracture eventually healed with a splint. Successful stable fusion of the shoulder in patients with brachial plexus injuries requires rigid fixation with a single 4.5 mm reconstruction plate, protection of bone healing with a triangular abduction brace for 12 weeks and functional trapezius, levator scapulae, serratus anterior and rhomboid muscles for optimizing the functional result. PMID- 16381289 TI - Operative treatment of trochanteric fractures with dynamic hip screw--is perioperative blood transfusion needed? AB - The need for perioperative blood transfusion in elderly patients with trochanteric fracture scheduled for elective dynamic hip screw fixation has recently been questioned following reports on the association between allogeneic blood transfusion and post-operative infections. This retrospective study was undertaken to assess the amount of per operative blood loss and transfusion requirement in relation to pre-operative haemoglobin level in 198 patients with trochanteric fractures. The average per operative blood loss was 409 ml and it correlated well with the duration of the operation. More than half of the patients (52.5%) required blood transfusion and nearly three-quarters were anaemic prior to the surgery. Proactive pre-operative measures to optimize the patient's haemoglobin level and intra-operative minimization of blood loss are essential steps to obviate the need for perioperative allogeneic blood transfusion. PMID- 16381290 TI - Mid-term results of tibial plateau fractures. AB - We studied the factors influencing the mid-term outcomes of tibial plateau fractures treated conservatively (n=21) and surgically (n=27) from December 1994 to December 1997. Joint stability was an important prognostic determinant. In the surgical group, the most important factor was good anatomical reduction. Functional outcomes were comparable between the conservative and surgical groups. We concluded that conservative treatment is a valid option for fractures with minimal displacement and surgical treatment is justified for severely displaced or depressed fractures. Attention must be paid to the recognition and restoration of joint stability and articular surface congruency for a satisfactory outcome. PMID- 16381291 TI - Revision surgery for developmental dysplasia of the hip. AB - Revision surgery following failed open reduction for developmental dysplasia of the hip (DDH) is technically demanding. We reviewed 12 patients in our institution that required a revision surgery between January 1994 and December 2003. The aims of this study are to identify the causes of redislocation after the primary open reduction and to determine the clinical and radiological outcome after the revision surgery. The mean age at presentation for DDH was 31 months (range 1-84) and the mean age at primary open reduction surgery was 38 months (range 15-84) and the mean age at revision surgery was 69 months (range 21-180). The mean follow-up period after revision surgery was 20 months (range 3-84). All the revision surgery was performed via an anterior Smith Peterson approach. The most common cause for redislocation was inadequate exposure and failure to release the obstructing soft tissues around the hip. The bony factors for redislocation included failure to perform a femoral shortening and excessive derotation of an anteverted femoral head combined with a Salter osteotomy. Three cases with posterior acetabular wall deficiency needed immobilization of the hip in extension as the hip tended to dislocate in flexion. Six patients had limb length discrepancy ranging from 1 cm to 4 cm and 50% developed avascular necrosis. Only 2 patients were asymptomatic and the majority had a limp and limitation of motion. All were pain free except one. In view of the technically demanding surgery and poor results after revision, the surgeon should recognize the pathology and ensure that the primary procedure achieves a stable and concentric reduction. PMID- 16381292 TI - Early results of Rotating Platform total knee replacement. AB - We report on our early experiences with the Press Fit Condylar Rotating Platform (PFC-RP) total knee replacement prosthesis at University Malaya Medical Centre. This new prosthesis was introduced to Malaysia in 2001. It combines the rotating platform technology of Low Contact Stress (LCS) and the modularity of Press Fit Condylar (PFC) system. We performed 18 knee replacements using cruciate retaining prosthesis without patellar resurfacing in 10 patients and followed them up over a period of 16-month. A new operative strategy was developed to match the flexion and extension gaps. The pre- and post-operative assessments were made according to the American Knee Society Scores (AKSS). Post-operative pain relief, range of motion, knee score and functional score showed promising early results. The average knee rating improved from 28 to 91 (range 75-94). Average functional assessment improved from 26 to 82 (range 75-90). The radiographic assessments show correction of an average pre-operative varus of 140 to post-operative valgus of 50 (range 4-70). Early infection occurred in two knees. Both responded to aggressive debridement and antibiotics. Stiffness was encountered in one patient. There were no spinouts of the insert. We are satisfied that the implants are performing well at the one-year mark and will continue monitor this cohort. PMID- 16381293 TI - Reconstruction of the heel defect with in-step island flap. A report of four cases. AB - Soft tissue loss of the heel is difficult to treat as it may cause significant morbidity particularly recurrent ulceration and subsequent chronic osteomyelitis. Reconstruction of such defect with local flap can provide good result. We report our experience in treating four patients with heel defect using the in-step island flap. The flap, which is based on the medial plantar neurovascular pedicle, provides a sensate durable tissue required for weight-bearing and normal gait. PMID- 16381294 TI - Severe open fracture of the tibia associated with massive soft tissue loss: limb salvage revisited. AB - The choice between limb salvage and primary amputation in a severely injured limb is at time difficult. A case of severe Gustilo type-IIIB open fracture of the tibia with massive soft tissue loss is presented to highlight the immediate and definitive treatment undertaken to preserve the limb. PMID- 16381295 TI - Progressive supra-axial cervical instability in Down syndrome: a case report emphasising the role of extended short-segment fusion. AB - We report a case of upper cervical instability associated with Down syndrome to highlight its potential progression to inflict cord compression and the rationale for surgical decompression and extended short segment occipito-axial fusion. PMID- 16381296 TI - Traumatic hip dislocation in late pregnancy: a case report. AB - Hip dislocation in pregnancy is an uncommon injury. We report a case of traumatic hip dislocation in the third trimester of pregnancy to highlight potential problems associated with its treatment. The rationale for choosing the preferred treatment options is discussed. PMID- 16381297 TI - Simultaneous dislocation of the metacarpophalangeal and subluxation of the carpometacarpal joints of the thumb in a child. AB - Double dislocations of joints of the thumb are uncommon injuries. We report an unusual case of simultaneous dislocation of the metacarpophalangeal (MCP) and subluxation of the carpometacarpal (CMC) joints of the thumb in a child to illustrate problems related to their treatment. PMID- 16381298 TI - Fillet flap: a successful procedure to preserve the optimum length of below-the knee-amputation stump in congenital constriction band of the leg. AB - The use of 'spare part' tissue from an amputated segment to cover a stump with soft tissue deficiency is a viable option for augmenting the shape of the stump and preserving functional length of the stump without additional donor morbidity. The procedure requires no microsurgical skill if the tissues are used as pedicle fillet flaps transfer. This concept of fillet flap makes the amputation distal to an unfavourable definitive amputation site such as in congenital constriction band possible. We report herein a case of amputation distal to constriction ring to illustrate the application of fillet flap in reconstruction surgery. PMID- 16381299 TI - Nurses can now initiate hormonal contraception. PMID- 16381300 TI - Disclosure of confidential information. Amendments to the Code of Ethics of Nurses. PMID- 16381301 TI - 2005-2006 General Supervision Program. PMID- 16381302 TI - [Physician's competence and obligations at the place of subject's death outside the health-care facilities]. AB - The valid law does not unequivocally define a physician's competence and obligations at the place of a subject's death outside the health-care facilities. The scheme creates a general instruction for the medical personnel to be used in cases of death outside health-care facilities. It is aimed at a better understanding of the problem, and it is supplemented with relevant legal regulations. The scheme is divided into 3 horizontal levels, the death being placed on the first level, the decision about autopsy on the second level, and the burial on the third level. These three levels are interconnected vertically, in order to represent the chronological sequence of individual steps and the respective decisions made by the examining physician. The schematic representation of the problems of death outside health-care facilities resulted in several lines of action, all that with regard to the medical and forensic aspects of the death. In every line of action, the work of the examining physician's is summarised. PMID- 16381303 TI - History of forensic medicine--the third part. The development and history of forensic medical science in Middle Europe. PMID- 16381304 TI - [Neural stem cells and their significance in regeneration processes in the nervous system]. AB - This review includes the literature data and the results of authors' own investigations on identification, cultivation and perspectives of therapeutic application of human and animal regional stem cells. Proliferation of pluripotent stem cells is observed in subventricular area of lateral ventricles and subgranular layer of dentate fascia of hippocampal formation of adult brain of man and animals. Data on the hierarchical organization of gene networks in the regulation of individual development may point to a possible functional role of repeating mini- and microsatellite DNA sequences in stem cell differentiation. The variants of application of human bone marrow as a source of stem cells for repair of damaged brain tissues are considered. It is established that heat shock proteins block the formation of glial scar during neurotransplantation. The viability of stem cells during transplantation may be improved by insertion of genes for neurotrophic growth factors into a genome of transplanted neurons. PMID- 16381305 TI - [Neocortex formation in mice developing after prenatal serotonin depletion]. AB - The objective of this study was a detailed investigation of structural changes taking place in murine neocortex during its formation and stratification after prenatal serotonin depletion. The study was carried out in murine embryos of F1 (C58BL/CBA) hybrid strain. For depletion of endogenous serotonin in mice, pchlorophenylalanin, an inhibitor of tryptophan hydroxylase - a key enzyme of serotonin synthesis, was used. Brain of the offspring was studied on postnatal days 1, 5 and 10 (n = 10-15 for each time point). Intact animals of respective developmental stages were used as a control. The study demonstrated that prenatal inhibition of serotonin synthesis resulted in malformation of all neocortical layers, disorders of neuronal growth, development and differentiation, changes in their shape and dimensions. During postnatal development, the loss of a significant numbers of cells was observed in the brain structures studied of serotonin-depleted animals. PMID- 16381306 TI - [Neuronal organization of thalamic nucleus reticularis in adult man]. AB - The neuronal content of human thalamic nucleus reticularis was studied in serial sections cut in sagittal and frontal projections and impregnated with silver nitrate using Golgi method. The neuronal content of human thalamic nucleus reticularis was found to be more diverse than previously reported in animals and man. Besides two types of sparsely-branched long-dendritic spineless R1 and R2 neurons, this nucleus contained spiny cells. Medium and small-sized sparsely branched short-dendritic neurons and densely-branched spiny cells were demonstrated. The principle of organization of human thalamic nucleus reticularis is described. PMID- 16381307 TI - [Hypothalamic neurosecretory cell apoptosis during stress in mice at different stages of ontogenesis]. AB - The aim of this work was to detect the participation of cycloferon in apoptosis control of cells of hypothalamic neurosecretory centers in young and old mice subjected to immobilization stress. Apoptotic cells were detected using ethidium bromide staining. Optic density of antiapoptotic Bcl-2 protein was determined in the immunoreactive cells of supraoptic and paraventricular nucleus (PVN). It was observed that cycloferon reduced activity of apoptosis in hypothalamic neurosecretory centers via Bcl-2-independent pathway. Cycloferon administration prior to stress had no influence on the number of apoptotic cells, except those ones in PVN of old animals, which demonstrated an apoptosis inhibition. PMID- 16381308 TI - [DNA, proteins and lipids distribution in the cells of olfactory bulbs of rats of various ages]. AB - Using luminescent and absorption stains, specific for DNA (acridine orange, ethidium bromide), proteins (silver nitrate) and lipids (sudan III), the distribution of these substances was studied in the sections of rat olfactory bulbs, fixed by paraformaldehyde. DNA prevalence was found in glomerular cell layer as compared with the mitral one. Large amount of DNA was detected in granular cell layer, underlying the mitral one. The peculiarities of cellular layers of olfactory bulbs of 2-day-old rats were compared with those ones in 1 month-old animals. In rats of different ages, the differences were found in DNA content and distribution between layers, while no differences were detected in total protein and lipids. In 2-day-old rats glomerular underdevelopment was demonstrated. PMID- 16381309 TI - [Structural changes of the retina and corpus vitreum in the model of proliferative vitreoretinopathy in alloxan diabetes]. AB - The present study was conducted to determine the role of blood mononuclear cells in a morphogenesis of proliferative vitreoretinopathy (PVR) and to detect the peculiarities of PVR development in hyperglycemia. Experiments were performed in 75 mature male Wistar rats. In animals of group 1, the model of PVR was reproduced by injection of mononuclear cells into the corpus vitreum. In animals of group 2, alloxan diabetes was induced 6.5 weeks prior to intravitreal injection of mononuclear cells. The material obtained was studied using the methods of light and electron microscopy. Development of PVR by intravitreal injection of mononuclear cells in diabetic rats was found to be associated with more severe destructive and proliferative changes. Considering the integrative role of blood mononuclear cells in the intercellular networks, it is suggested that these cells are involved in the pathogenesis of PVR. PMID- 16381310 TI - [Smooth myocytes of cerebral arteries at the emergency stage of hypertension]. AB - The objective of this study was to analyze the structural modifications of smooth myocytes (SM) in the tunica media of cerebral arteries at the emergency stage of hypertension. SM were isolated by the method of targeted alkaline dissociation from the tunica media of medial cerebral artery in 22 pups (10 animals comprising the control group). Emergency stage of hypertension was induced by an experimental aortal coarctation. Mathematical analysis of parameters of arterial pressure, inner diameter and tunica media thickness is indicative of a reduction of permissible tensile strength of cerebral arteries. The transformation of vascular SM was demonstrated that included the increase in their dimensions and the decrease of their nuclear/cytoplasmic ratio. Destructive and compensatory adaptive SM changes (increase of nuclear DNA content and of the proportion of binuclear forms) were detected. These changes in the structure of arterial SM population in combination with the decrease of permissible tensile strength, may contribute to an increased risk of vascular wall rupture and the development of an acute dysfunction of cerebral circulation. PMID- 16381311 TI - [Analysis of the changes of stromal precursor cell numbers in the thymus and the spleen of animals of different age groups]. AB - The objective of this study was to analyze the species differences in the numbers of stromal precursor cell (CFU-f), their cloning efficiency (CFE-0 and their dynamics in different organs during aging, using the mathematical gradient decrease method. Age changes of CFU-f numbers and of their CFE-f were studied in the thymus and the spleen of mice and guinea pigs. The study was performed using CFU-f cloning in monolayer cultures. CFU-f numbers and CFE-f were found to decrease with aging both in the thymus and the spleen of mice and guinea pigs. However these changes were different in each species and were variable in different organs of the animals of the same species, which, probably was associated with the physiological characteristics and aging peculiarities of the animals of different species and with the functional role of organs studied. The process of reduction was more significant in the thymus of guinea pigs and mice - the numbers of CFU-f were decreased 75- and 12-fold, respectively. Since it is known that the population of CFU-f in the thymus and the spleen includes inducible osteogenic precursor cells, the data obtained indicate the possibility of a reduction in numbers of this category ofstromal precursors, that could be one of the reasons of osteoporosis of aging. The application of a mathematical analysis using the gradient decrease method allows to predict the time-course of age changes and to evaluate the dynamics of CFU-f numbers and of CFE-f in association with organism aging. PMID- 16381312 TI - [Inguinal lymph nodes of monkeys subjected to 8-days-long immersion in water]. AB - The morphometric study of cytoarchitecture of the structural and functional components of lymph nodes of male monkeys (Macaca mulatta) aged 4-5 years was performed after 8-days-long immersion in water. Activation of lymphocyte mitotic activity in lymphoid nodules (B-dependent zones) was demonstrated in association with an accumulation of small lymphocytes in them and an increase of macrophage reaction. Accumulation of mature forms of plasma cells was detected in all the structures of lymph nodes. It was also found that 8-days-long immersion in water resulted in remodeling of the vessels of lymph node microcirculatory bed, which was accompanied by changes of the endothelium in blood vessel wall and by disturbances of lymph drainage, that lead to accumulation of lymphocytes in lymph node sinuses. PMID- 16381313 TI - [Comparative characteristic of structural-functional changes of the spleen in the offspring of female rats with chronic liver pathology of various etiology]. AB - The aim of the study was a comparative analysis of structural-functional changes of the spleen in the offspring of female rats with chronic liver pathology of various genesis. In adult female Wistar rats various forms of liver lesions were modeled: toxic, autoimmune, cholestatic. The spleen of the offspring of the experimental animals with chronic liver disease was studied at different time points of postnatal ontogenesis. The increased numbers of Ig-producing cells and splenic karyocytes were detected in association with an increased surface area of the section, occupied by lymphoid nodules. Activation of proliferation and differentiation of B-lymphocytes was found together with an increase of the surface area of the section, occupied by B-dependent zones, indicating the increased intensity of humoral immunity in the offspring of female rats with chronic pathology of hepatobiliary system of various genesis. PMID- 16381314 TI - [Endocrinocytes of the human bulbourethral glands]. AB - Structure, topography and numbers of endocrinocytes in bulbourethral glands of mature men were studied using immunohistochemical demonstration of chromogranin A. Chromogranin-positive cells were found to be predominantly localized in the epithelium of excretory ducts, while they were sparse in the terminal secretory portions. Endocrinocytes in bulbourethral glands were shown to possess argyrophobic properties and to be stained with antibodies against common cytokeratin. The possibility of epithelial histogenesis of bulbourethral gland endocrinocytes is discussed. PMID- 16381315 TI - [Oxidative metabolism of uterine smooth muscular tissue in norm and neoplastic growth (clinical and experimental studies)]. AB - Complex echoscopic, histological and EPR- (electron paramagnetic resonance) spectroscopic study of uterine smooth muscular tissue was performed using normal and neoplastic samples obtained from women of reproductive age as well as from experimental animals (guinea pigs) with normo- and hyperestrogenemia. It was found that as compared with normal myometrium, the proliferating uterine myoma had an extensive peripheral vascularization of myomatous nodule with a decreased resistance index, which is a marker of myocyte proliferative activity in the myomatous nodule. These data were supported by histological findings in material obtained at operations, which demonstrated the signs of proliferative growth. Using EPR, it was shown that uterine myoma was characterized by an estrogen dependent intensification of the processes of free-radical oxidation, which correlated with a degree of hormonal changes. Accumulation of free-radical oxidation activators and of ions of ferritin-unbound iron in the tumor tissue is indicative of the intensification of proliferative activity of the cells of uterine myoma and is one of the risk factors of neoplastic growth. Hyperestrogenemia, characteristic for the myoma development, is one of the reasons for NO synthesis activation, which, in the oxidative stress, is transformed into cytotoxic peroxinitrite, contributing to further intensification of an oxidative stress and malignant transformation of tissues. PMID- 16381316 TI - [Peculiarities of repair osteogenesis during the transverse traction of the longitudinal diaphysis fragment under the conditions of intraosseus vessel preservation]. AB - The application of technique for obtaining the tibial fragment (split) with a preserved vascular network of the nutrient artery and its branches, that participate in the blood supply of this bone area, permits to detect significant osteogenetic potential of both endosteum and undifferentiated bone marrow cells. Under these conditions it was possible to increase the bone thickness 2 to 3 times in short time. The regeneration process in this case is characterized by a rapid course and formation of large massive of cancellous bone. The results of this study suggest that an adequate blood supply, stable fixation of bone using Ilizarov apparatus and the chosen rate of split displacement create the favorable conditions for the active osteogenesis. PMID- 16381317 TI - [Experimental morphological analysis of perichondrial chondrogenesis]. AB - Microscopic structure of perichondrium of rabbits of different age groups was studied under normal conditions and after its ortho- and heterotopic transplantation in several variants. The results obtained showed that after isolation from its normal environment and placement into unusual conditions of survival, perichondrium maintained its capacity for growth; its cells were found to divide and differentiate actively eventually forming regular hyaline cartilage in a manner similar to that taking place during normal ontogenesis. Perichondrial chondrogenesis occured in all cases of free perichondrial transplantation, provided the experimental conditions assured the optimal vascularization of transplant bed and there were no inerference with the realization of perichondrial histoblastic potential. The most intensive and full-scale chondrogenesis was obtained in the experiments that provided the expansive growth of transplant due to availability of a free space above it. Under these conditions, the proliferation of chondroblasts occured similar to that taking place in normal articular cartilage, outer surface of which faces the free space of articular cavity. These experimental data were supported by the clinical practice, where perichondrial transplantation is successfully used for the treatment of osteoarticular pathology. PMID- 16381318 TI - [Minor congenital malformations: prognostic indicator of ill-being of an infant born by alcoholic mother]. AB - This work presents the results of a comparative analysis of risk factors and phenotypical characteristics of 228 children with fetal alcohol syndrome and of 347 children with hypoxicischemic encepalopathy. The study of a phenotype detected that an average number of minor congenital malformations in children with fetal alcohol syndrome was equal to 12.2 while in comparison group it was equal to 7.3. High prevalence of stigmata was associated with an increased frequency congenital malformations of internal organs and mental disorders in children by the age of 4. PMID- 16381319 TI - [Modification of histogenetic processes in rat nervous tissue after dexamethasone administration during prenatal development]. AB - In experiments performed on rats, using the methods of confocal microscopy and immunohistochemistry, it was detected that administration of a single dose of synthetic corticosteroid dexamethasone during the period of increased sensitivity of developing nervous system (embryonic day 13) resulted in modification of histogenetic processes in the brain, resulting in the changes of its structural and functional characteristics during postnatal period of ontogenesis. PMID- 16381320 TI - [Primary sensory neurons in the central nervous system]. AB - This paper summarizes the results of author's personal observations and the literature data on exteroceptive and some interoceptive neurons of the brain and spinal cord, such as Hesse's intraspinal ocelli, light-sensitive epiphyseal and ependymal neurons. Also described are the light-sensitive ganglion neurons of invertebrates, true bipolar sensory neurons inside the spinal cord, primary chemo and thermosensitive neurons and sensory unipolar neurons, accompanying four thin Motavkin "central nerves", perforating spinal cord meninges and terminating with bush-like receptors near blood vessels or close to canalis centralis ependyma. The data on all known vertebrate intracerebral and intraspinal interoceptors are summarized in a unified scheme. The hypothesis is formulated, according to which animal and human central nervous system possesses its own sensory innervation, comparable to that one of the other organs, and contains local primary sensory neurons and their asynaptic dendrites, which may be divided into two groups: interoceptive and exteroceptive. PMID- 16381322 TI - [Bulbourethral glands]. PMID- 16381321 TI - [Sensory innervation of the brain--myth or reality?]. AB - This paper contains a critical analysis of conceptions of the sensory innervation of the spinal cord and the brain, presented in the articles by O.S. Sotnikov (issues 2 and 3 of this journal). The criteria for identification of intracerebral primary sensory neurons are examined, as well as the possible role of neuronal cilia. The significance of asynaptic dendrites and bipolar cells is discussed. The importance of an adequate usage of some terms and the advisability of a combined morphological and physiological analysis for the study of sensory intracerebral exteroceptors is emphasized. PMID- 16381323 TI - [Role of Professor August Rauber's Institute of Anatomy in the development of anthropology in Tartu (former Imperial Derpt/Yuriev) University in 1886-1911]. PMID- 16381325 TI - NCQA, health plans promote movement back to HMOs. PMID- 16381326 TI - Health plan cost trends for 2006 show HMO metrics lowest. PMID- 16381327 TI - Are 'persistently high users' burdening your primary care practice? PMID- 16381328 TI - Michigan plans look for savings in Medicaid managed care. PMID- 16381329 TI - Capitation shrinks in multispecialty IDS practices, but primary care bucks trend. PMID- 16381330 TI - Evaluation of the child with developmental delay. PMID- 16381331 TI - A clinical analysis of children with developmental delay. AB - The purpose of this study was to identify the functional domains and etiological risk factors of children with developmental delay (DD). During a 41-month period, a retrospective chart review was carried out on all 1192 patients who visited pediatric clinics for evaluation of developmental delay or behavioral problems. Etiological risk factors or associated disorders were identified after the completion of clinical evaluation and a series of laboratory investigations. Clinical psychologists examined the functional domains of those patients who underwent developmental and psychological assessments. Those children who met the criteria of DD were subdivided into six functional subtypes, including cognitive, motor, language, social adaptation, global and non-specific developmental delay. A total of 1192 children were diagnosed as DD, 86.7% had cognitive DD, 58.4% had language DD and 47.9% had motor DD. Of 1192 children, 831 were male, and the ratio male to female was 2.3 to 1. Only 20.5% (244) of children had identifiable etiological risk factors and 35.6% (424) had associated disorders. Results showed DD was more prevalent in males, and cognitive delay was the most common subtype. Identifying the etiological risk factors of DD remains difficult because of its heterogeneity. PMID- 16381332 TI - Prognostic predictors of neurodevelopmental outcome or mortality in very-low birth-weight infants. AB - Survival rate of very-low-birth-weight (VLBW) infants has much improved in recent years. However, neurodevelopmental outcomes are still of the most concerned to such infants' parents. In order to survey survival rate, neurodevelopmental outcome and prognostic factors in these infants, we did a retrospective study of VLBW infants who were admitted to neonatal intensive care unit of Taipei City Hospital, Branch for Women and Children from 1st January 2001 to 31th October 2004. We reviewed their medical records and analyzed the data. A total of 68 cases were enrolled in this study, and overall one-year survival rate was 70.3% (45/64). The unfavorable outcome group had lower birth weight, lower gestational age, lower Apgar scores at 1-minute and 5-minute, higher rate of vaginal delivery, more respiratory distress syndrome (RDS), and higher IVH grades (all p value < 0.05). However, with multiple logistic regression analysis, only low birth weight and higher IVH grades were the significant predictors of unfavorable outcome. Univariate analysis showed that RDS increased but Cesarean section (C/S) decreased the risk of severe IVH. According to the result, we can make the conclusion that the survival rate of VLBW infants in our hospital was similar to that in other medical centers in Taiwan. In order to prevent severe grades of IVH and subsequent unfavorable outcome in VLBW infants, prenatal therapies to prevent RDS and perhaps choice of C/S as delivery mode are the best strategies. PMID- 16381333 TI - The rescue of acute fulminant myocarditis by extracorporeal membrane oxygenation in pediatric patients. AB - The severity of acute myocarditis varies between subclinical to lethal. For fulminant myocarditis, the mortality rate can be as high as 75% when shock occurs. Extracorporeal membrane oxygenation (ECMO) support to revert this fulminant course has been introduced into the management in recent years. Six such pediatric cases rescued with ECMO in our hospital over a 3-year period were reviewed. Femoral venoarterial cannulated ECMO was undertaken in 5 patients and right atrium and ascending aorta cannulated ECMO in one. All patients had histories and clinical findings consistent with fulminant myocarditis. Median age was 12 years old (range 9 to 14 years). Median duration of ECMO support was 61.5 hours (range 36 to 90 hours). Three patients survived. Two of them complicated with deep vein thrombosis or peripheral neuropathy. The survivors showed normal cardiac function after follow-up periods of 0.8-4.3 years. Early recognition and immediate establishment of an ECMO circuit are crucial to rescue a patient with fulminant myocarditis. Every effort to avoid the complications associated with ECMO should then be stressed. PMID- 16381334 TI - Maternal acupuncture effects on surfactant and antioxidant enzymes in preterm rat lungs. AB - The objectives of this study are to evaluate the effects of maternal acupuncture treatment on lung maturation in preterm rats. Two stainless-steel needles were inserted into the Tsu-San-Li locus in the right hind leg of timed pregnant Sprague-Dawley rats for 30 min. One-day acupuncture-group mothers received electroacupuncture on day 18 of gestation. Two-day acupuncture-group mothers received electroacupuncture on days 17 and 18 of gestation. Control-group mothers received acupuncture at a site not contained in the Atlas of Human Acupuncture Points on day 18 of pregnancy. On day 19 of gestation, pups in all dams were delivered by cesarean section. Maternal 2-day acupuncture treatment significantly increased total phospholipids in fetal lung tissue when compared with control and 1-day acupuncture-treated groups. Two-day acupuncture-treated fetuses had higher saturated phosphatidylcholine level in lung tissue although the difference did not reach statistical significance. Two-day acupuncture-treated fetuses had significantly lower superoxide dismutase, catalase, and glutathione peroxidase activities than did the control and 1-day acupuncture-treated fetuses. We conclude that maternal acupuncture treatment affects surfactant and antioxidant enzyme development in contrasting ways and may have both beneficial and potentially harmful effects on different aspects of lung development. PMID- 16381335 TI - Prescribing patterns of anti-asthma drugs in pediatric patients. AB - Guidelines from the Global Initiative for Asthma (GINA) mention several medications for the treatment of asthma. These medications include oral and inhaled beta-2 agonists, oral and inhaled corticosteroids, xanthines, leukotriene receptor antagonists, and their combinations. In addition to asthma, these drugs are commonly prescribed to treat other respiratory diseases, such as acute bronchitis, chronic cough, lower respiratory infection, or even bronchopneumonia. We analyzed differences in prescribing patterns between pediatric patients with and those without asthma, as coded in the claim records from the National Health Insurance Research Database. Oral beta-2 agonists were the most frequently monotherapy in both groups of patients (52.6-77.6% vs 62.8-84. 8%). Oral beta-2 agonists combined with xanthines or oral corticosteroids combined with an oral beta-2 agonist were the most frequent combination therapies in both groups. Inhaled corticosteroids were used in 3.1-11.0% of patients with asthma; the rate varied by patient age. In conclusion, prescribing patterns were similar in pediatric patients with and those without asthma. PMID- 16381336 TI - Premature with trichomoniasis: report of one case. AB - We report a case of an extremely-low-birth-weight premature infant with Trichomonas vaginalis infection of the vagina and urinary tract. Her mother is also infected with Trichomonas vaginalis but is asymptomatic. The patient's illness started as an asymptomatic pyuria and later on at early infancy developed profuse malodorous vaginal discharge. Her vaginal discharge was positive for Trichomonas vaginalis, both on wet mount and Papanicolaou smear, and the infection responded well to treatment with metronidazole. Infants with recurrent vaginal discharge presenting beyond the neonatal period may be attributed to T. vaginalis infection. PMID- 16381337 TI - Hepatopulmonary syndrome: report of one case. AB - Hepatopulmonary syndrome consists of a triad of chronic liver disease, hypoxemia, and intrapulmonary vascular dilatation in the absence of primary cardiac or pulmonary diseases. Though this complication has been described in as many as 4 29% of patients with liver disease, its clinical manifestations are often subclinical. The standard diagnostic tests for hepatopulmonary syndrome are the applications of contrast echocardiography and 99mTc lung perfusion scan. In this report, we describe a 19-year-old female with biliary atresia and subsequent liver cirrhosis, who also had the complication of hepatopulmonary syndrome since adolescence. We conclude when the symptoms and signs of hypoxemia appear in a patient with chronic liver disease, the differential diagnosis of hepatopulmonary syndrome should be kept in mind. PMID- 16381338 TI - Identification of OCRL1 mutations in two Taiwanese Lowe syndrome patients. AB - The oculocerebrorenal syndrome of Lowe (OCRL) is a rare X-linked multisystem disorder characterized by congenital cataracts, mental retardation, and renal tubular dysfunction. The OCRL1 gene responsible for Lowe syndrome has been mapped to chromosome Xq24-q26. We analyzed two Taiwanese OCRL patients and their families. In Case 1, a splicing mutation (889-11 G --> A) was identified in intron 10 of the OCRL1 gene. The mother is a heterozygous carrier. The 889-11 G - > A mutation results in an abnormal splicing and predicts premature termination of translation. In Case 2, a novel de novo missense mutation (1373G --> A, P458H) was identified in exon 14 of the OCRL1 gene. The missense mutation predicts a substitution in a domain highly conserved among the inositol-5-phosphatase family. PMID- 16381339 TI - Peritoneal metastasis in osteosarcoma: report of one case. AB - We describe a rare case of peritoneal metastasis in osteosarcoma. A 14-year-old boy was diagnosed with osteosarcoma of the femur. Although initial treatment with surgical resection and chemotherapy was successful, a large inoperable pulmonary metastasis was detected 27 months after diagnosis of the primary tumor. At 38 months, whilst still receiving salvage chemotherapy, malignant ascites with peritoneal metastasis was detected. The patient died two months later from disease progression. This is probably the 6th case and youngest patient with peritoneal metastasis of osteosarcoma reported in the literature. For early diagnosis, patients with a history of osteosarcoma having persistent abdominal symptoms or signs should be offered a CT scan of the abdomen, and examination of the ascitic fluid when present. With advances in treatments, unusual sites of metastasis may become more prevalent. PMID- 16381340 TI - Axillary supernumerary breast in a female adolescent: report of one case. AB - Polythelia and polymastia usually occur along the embryonic milk lines extending from the axilla to the groin. Polymastia in female patients has been reported to manifest during pregnancy or lactation. We report a 14-year-old adolescent with axillary supernumerary breasts. She had painful axillary swelling during menstrual period. The mass in the right axilla was excised with pathologic report of supernumerary breasts with fibrocystic change. When a mass is located along the milk line, the possibility of the presence of breast tissue should be considered. PMID- 16381341 TI - Poland syndrome in a neonate: report of one case. AB - Poland syndrome is an infrequent disease that presents in newborn infants and features an incidence of about one in 7,000 to one in 100,000 live births. The clinical features include an absence of the costosternal portion of the pectoralis major muscle, hypoplasia and/or aplasia of the ipsilateral breast or nipple, hypoplasia of subcutaneous tissue, abnormalities of the rib cage and upper extremity anomalies. We present a male newborn suffering Poland syndrome who featured a depressed left chest wall with respiratory distress. Ultrasonography of the chest revealed the absence of a section of pectoralis major muscle over the left chest wall, although no abnormality was found over the ipsilateral upper extremity. The patient was regularly followed up and showed no limitation to the range of motion of the left upper extremity at the age of four months. PMID- 16381342 TI - Short staffing and iatrogenic stressors: can we survive our own doings? AB - If there was ever a time when the nursing profession was under the spotlight and gaining national and local attention, this is it. There are those whose efforts will help with nurse recruitment and retention. There will be increased funding for scholarships and other financial inducements for nurses. Regulators are looking at mandated nurse-patient ratios and examining mandatory overtime for nurses. External change agents can--and are--influencing the state of nursing. There are changes that need to be made within nursing. Systems, role, and environmental change is best directed by nurses. We are called to be internal change agents within nursing with an eye toward seriously re-examining how we create our own stress. Using a 1970's team model for eight-hour shifts is redundant with 12-hour scheduling models in place. Recalling the scarcity of vital information in days gone by (i.e., blood gases were drawn but once a day as an adjunct to intense nursing observations), today there is a plethora of unending clinical data, such that limits need to be established. Failing to sufficiently address the human condition in a coordinated and effective manner, given the cost of care and the rising expectations of consumers is a constant challenge if we fail to address the soundness of our operational systems. Systems must be kept current with the times. For instance, how we blend temporary workers with "core resource" nurses remains largely unanswered. We must systematically address these issues, or be subject to the stress caused by the failure to do so. Nurse leader--and effective nurse followers--should use the spotlight to demand a re-hauling of our ineffective practices. Not in the spirit of work reengineering aimed at cost savings alone, but pragmatically to address the essence of the nurse-patient relationship and to create a savory environment for nursing professionals. PMID- 16381343 TI - West Nile Virus. AB - West Nile Virus (WNV), a mosquito transmitted disease, made its first appearance in North America in 1999 and quickly spread westward. It arrived in Kansas in 2002, re-emerged in 2003, and is expected again in 2004. The natural cycle is from infected bird to mosquito, with humans and horses as accidental (dead-end) hosts. WNV is usually asymptomatic in humans, but can cause a wide spectrum of disease. There is currently no human vaccine and treatment is symptomatic only. A best preventive measure combines personal protection to prevent mosquito bites with reducing the number of mosquitoes. PMID- 16381344 TI - Diarrheal illnesses: a public health perspective. AB - Diarrheal illnesses remain among the leading causes of morbidity in the United States. Approximately five million diarrheal cases occur annually (Chin, 2000; Ostroff & Leduc, 2000), with an estimated incidence of one diarrheal episode per person per year (Aranda-Michel & Giannella, 1999). Though the causes of diarrheal illnesses vary, infectious agents account for a majority of cases (Aranda-Michel & Giannella, 1999; Chin, 2000; Ostroff & Leduc, 2000). Most diarrhea-causing infectious agents are transmitted through food, water, or person-to-person via the fecal-oral route and are the cause of numerous diarrheal outbreaks. The risk for exposure to such pathogens within the general population is universal; however, persons in pediatric, geriatric, and other immunocompromised populations are at increased risk for subsequent illness and complications (Centers for Disease Control and Prevention, 2001; Ostroff & Leduc, 2000). Moreover, many persons with diarrheal illness do not seek medical care and self-treat with over the-counter antidiarrheal agents, which have potentially serious side effects among high-risk individuals. The public health impact of diarrheal illness is apparent and emphasizes the need for early diagnosis and appropriate treatment, timely notification of illness with public health implications, and coordination between healthcare professionals and public health officials to prevent and control the spread of infection. PMID- 16381345 TI - Hepatitis C: a public health response in Kansas. AB - The hepatitis C (HCV) infection prevalence in Kansas is concentrated in populations with increased risk factors for acquiring the virus. Testing the appropriate population and providing counseling and medical referral to these high risk individuals is accomplished by offering HCV testing at HIV Counseling and Testing sites across the state. Such public health programs along with the medical community allow the at risk populations of Kansas an opportunity to improve their health and decrease the spread of HCV PMID- 16381346 TI - Reducing surgical site infection through process improvement initiatives. PMID- 16381347 TI - 2005 Presidential Address. PMID- 16381348 TI - Laparoscopic transhiatal surgery of the esophagus. AB - OBJECTIVE: Esophagectomy is an operation with high morbidity and mortality. Its adoption as a minimally invasive operation worldwide has been slow, but the potential benefits of reducing the trauma of surgery need to be considered. Our 30-month experience with transhiatal esophagectomy in a district general hospital is presented herein. METHODS: Patients were considered for surgery after radiological staging had excluded inoperable disease. Laparoscopic staging was initially performed. Patients with tumors of the esophagus and high-grade dysplasia in a Barrett's esophagus were included. RESULTS: Twenty-nine patients were referred for consideration for resectional surgery. Nine underwent outpatient laparoscopy only. Twenty patients (age range, 34 to 78, 15 males:5 females) underwent resectional surgery. Seventeen transhiatal resections were completed, 2 were converted to open procedures, and 1 transhiatal resection of a benign tumor was performed. Median time of surgery was 415 minutes (range, 320 to 480) and blood loss was 300 mL (range, 200 to 350). The median length of post operative ventilation and critical care stay were 1 (range, 1 to 4) and 4 (range, 2 to 8) days. Median duration of hospitalization was 17 days (range, 10 to 28). Thirty-day mortality was 0; 1 patient who was converted to an open procedure died after a cerebrovascular event on day 34. CONCLUSION: A zero mortality rate for laparoscopic resection and a low-morbidity rate compare well with morbidity and mortality in reported series using this method and open surgery. Laparoscopic transhiatal esophagectomy is an advanced, complex procedure that can be performed safely in a district general hospital setting. PMID- 16381349 TI - Laparoscopic sigmoid colectomy for chronic diverticular disease. AB - BACKGROUND: The feasibility of laparoscopic sigmoid colectomy for diverticular disease has now been well established. We report herein our experience with laparoscopic sigmoid colectomy in 100 patients who underwent laparoscopic colectomy for chronic diverticular disease. METHODS: A retrospective review was performed of a 7-year period from January 1995 to June 2002. Chronic diverticular disease was treated with laparoscopic sigmoid colectomy in 100 patients. The setting was a community hospital. All cases were performed by 1 of 2 colorectal surgeons. All laparoscopic sigmoid colectomy patients received lighted ureteral stents placed preoperatively that were removed at the end of surgery. RESULTS: Mean age was 61.6 years. The male to female ratio was 38:62. The mean estimated blood loss was 138 mL, liquid diet was tolerated for 2.4 days, and hospital length of stay was 4.6 days. The mean operative time for laparoscopic sigmoid colectomy was 196 minutes. Relative complications for laparoscopic sigmoid colectomy are as follows: anastomotic leak in 2 (3.0%) patients, hematuria in 95 (95%) with an average duration for 3.1 days, urinary tract infection in 6 (6%), and ureteral injury in 1 (1%). The mean operating room charges in the laparoscopic sigmoid colectomy patients was dollars 9,643. CONCLUSION: We recommend laparoscopic sigmoid colectomy as the modality of treatment for chronic diverticular disease. Laparoscopic sigmoid colectomy appears to be a reliable, safe, and efficacious treatment modality for chronic diverticular disease. The operative time for laparoscopic sigmoid colectomy is decreasing as surgeons gain more experience. PMID- 16381350 TI - Early experience with laparoscopic pyloromyotomy in a teaching institution. AB - OBJECTIVE: Hypertrophic pyloric stenosis is a common pediatric surgical condition. A Ramstedt pyloromyotomy is performed either via laparotomy or laparoscopy. We report our first 25 cases of laparoscopic pyloromyotomy at an academic children's hospital. METHODS: From January 2002 through February 2003, we retrospectively reviewed our first 25 laparoscopic pyloromyotomies. All patients had documented hypertrophic pyloric stenosis by ultrasound criteria. Three incisions were made, one 5-mm umbilical port, one 3-mm right upper quadrant port, and a third left upper quadrant working stab incision. A 4-mm, 30 degrees scope was used in all cases. A longitudinal pyloromyotomy was performed using an arthrotomy scalpel. The pylorus was further separated with a laparoscopic Benson spreader. At the completion of the pyloromyotomy, the stomach was insufflated with air to identify any mucosal injury. RESULTS: Age range was 2.3 weeks to 8.4 weeks. Operating time has decreased from 70 minutes to 15 minutes. Two conversions to an open procedure were necessary, both during the first 10 cases. No mucosal perforations or incomplete pyloromyotomies have occurred. Feeds were started within 4 hours and advanced to goal. Time to discharge ranged from 12 hours to 30 hours. One patient developed umbilical cellulitis that was successfully treated with antibiotics. CONCLUSIONS: Laparoscopic pyloromyotomy is a safe, effective procedure for hypertrophic pyloric stenosis in a resident teaching environment. Laparoscopy permits excellent visualization, has comparable postoperative recovery, and superior cosmesis. PMID- 16381351 TI - Laparoscopic splenectomy in children. AB - BACKGROUND: Laparoscopic splenectomy is being performed more commonly in children, although its advantages are not clear. We sought to determine whether laparoscopic splenectomy was superior to open splenectomy. METHODS: The records of all pediatric patients undergoing splenectomy without significant comorbidities over a 12-year period were examined. The patients were divided into those undergoing laparoscopic splenectomy and those undergoing open splenectomy. Demographics, operative time, estimated blood loss, spleen size, length of stay, and total charges were compared between the groups. RESULTS: Eighty-one (58%) children underwent laparoscopic splenectomy, and 59 (42%) children underwent open splenectomy. The groups were similar in age and sex; hereditary spherocytosis was more common in the LS group. Operating time was longer in the laparoscopic splenectomy group (231 +/- 10 min vs 138 +/- 9 min; P<0.001), but blood loss and complication rates were similar. Twelve (15%) conversions were necessary primarily due to spleen size. Although children undergoing LS had a shorter length of stay (2.4 +/- 0.1 vs 4.1 +/- 0.3 days; P<0.001), they incurred higher charges (dollars 21199 +/- 664 vs dollars 15723 +/- 1737; P<0.002). CONCLUSION: Laparoscopic splenectomy is a safe procedure in children, resulting in shorter hospital stay, which may translate into earlier return to activity and a smaller burden on the child's caretakers. PMID- 16381352 TI - Laparoscopic inguinal hernia repair in children. AB - BACKGROUND: This study aimed to document the authors' experience with laparoscopic inguinal hernia repair in children. METHODS: Ninety-three hernia repairs were performed in 64 children. The neck was closed with a purse string suture by using 4-0 absorbable suture. RESULTS: Ninety-three indirect inguinal hernial sacs were closed in 64 children. Nine percent of children had an ectopic testis. The mean operating time for laparoscopic ring closure was 25 minutes (range, unilateral 21 to 35; bilateral, 28 to 50). The contralateral processus vaginalis was patent in 20% of children. In 24% of children, the final procedure was modified based on the findings of a dilated internal ring. A laparoscopic ilio-pubic tract repair was done in these cases. Laparoscopic mobilization, orchiopexy followed by ilio-pubic tract repair was done in 9% of children. Scrotal swelling occurred in one child. Hydrocoele occurred in one patient. Recurrence rate was 3.1%. CONCLUSION: Laparoscopic inguinal hernia repair in children can be offered, as it is safe, reproducible, and technically easy for experienced laparoscopic surgeons. Ilio-pubic tract repair may be added in cases with dilated internal ring. Recurrence following laparoscopic ring closure can be managed with laparoscopic ilio-pubic tract repair. The long-term follow-up of laparoscopic ilio-pubic tract repair is awaited. PMID- 16381353 TI - Laparoscopic repair of small bowel perforation. AB - OBJECTIVES: For years, limiting operative wound sepsis and its resultant morbidity in patients with small bowel perforations has been a major headache for surgeons. The present study was intended to extend the benefits of minimally invasive surgery to traumatic and typhoid small bowel perforations, in terms of assessing its feasibility and limiting wound sepsis. METHODS: Twenty-five patients (20 with typhoid ileal and 5 with traumatic small bowel perforation) presenting within 96 hours of the catastrophe were included in the study. All were explored laparoscopically, and the perforation repaired by intracorporeal suturing. RESULTS: Time of operation varied from 45 minutes to 92 minutes. Two patients developed port-site infection. None of the patients developed postoperative fistula, and no mortalities occurred. Postoperative hospital stay was between 7 days and 10 days. CONCLUSION: Benefits of minimally invasive surgery can be safely and efficaciously extended to select patients with small bowel perforation in terms of limiting sepsis-related wound complications. PMID- 16381354 TI - A randomized comparison of the early outcome of stapled and unstapled techniques of laparoscopic total extraperitoneal inguinal hernia repair. AB - OBJECTIVE: The need for stapling is a relative drawback of laparoscopic hernia repairs because it adds to the complications and costs. The safety of unstapled repairs as a viable alternative lacks validation, due to the dearth of analogous comparative trials. METHODS: Patients were randomized to undergo either stapled or unstapled total extraperitoneal hernia repairs. The groups were matched for age and the type of hernia repaired. Pain scores, intraoperative complications, postoperative complications, postoperative recovery, and long-term outcomes (ie, groin pain, paraesthesias, testicular atrophy, and recurrence) were studied. RESULTS: The incidence of complications, pain scores, pain trends, hospital stay, return to activity, and long-term outcomes were comparable. No recurrence has been noted at a median follow-up of 23 months in 63 hernias repaired in 49 patients. CONCLUSION: Unstapled laparoscopic hernia repair scores are equivalent to their stapled counterparts with respect to recurrence and complications. PMID- 16381355 TI - Laparoscopic cholecystectomy in the elderly. AB - OBJECTIVE: To evaluate the outcomes of laparoscopic cholecystectomy in elderly patients at a single institution. METHODS: A retrospective chart review was conducted of all patients > or = 65 years of age who underwent laparoscopic cholecystectomy over a 5-year period (January 1995 to December 1999). Four-trocar site laparoscopic cholecystectomy using the open Hasson technique were performed in all patients. The demographic data (age, sex), associated comorbidities, American Society of Anesthesiologist's (ASA) score, postoperative morbidity, mortality, and length of stay were recorded for each patient. Statistical analysis was done using Fisher's exact test and chi-square analysis. Statistical significance was defined as P < or = 0.05. RESULTS: The patient cohort included 46 patients with a median age of 71 years (range, 65 to 87). Seventeen (37%) patients were < or = 70 years of age, and twenty-nine (63%) patients were > or = 70 years of age. Twenty-two (48%) patients had ASA scores of > or = 3. Patients > or = 70 had significantly higher ASA scores. Eighteen patients > or = 70 years had ASA > or = 3 compared with 4 patients < or = 70 with ASA > or = 3 (P<0.05). Twenty-two patients > or = 70 and 8 patients < or = 70 required urgent surgery P<0.05). Fifteen (33%) patients presented with acute cholecystitis, and 31 (67%) patients presented with a greater number of chronic symptoms. Four (9%) patients had pancreatitis on presentation, and 6 patients underwent preoperative endoscopic retrograde cholangiopancreatography (ERCP). Two of these 6 patients also underwent sphincterotomy. Urgent surgery was performed in 30 (65%) patients. The mean operative time was 103 +/- 37 (SD) minutes. One (2%) conversion to open cholecystectomy was required. The mean postoperative stay was 7 days (range, 1 to 46). Fourteen (30%) patients had only a 1-night postoperative stay. Patients > or = 70 had significantly longer postoperative stays. Nine patients > or = 70 and only 1 patient < or = 70 stayed in the hospital for more than 7 days. Postoperative complications were noted in 6 (13%) patients, most of which were chest infections. Five patients > or = 70 and only 1 patient < or = 70 developed postoperative complications. No mortalities occurred. CONCLUSION: Laparoscopic cholecystectomy is safe and feasible in elderly patients. Patients > or = 70 years seem to have a longer postoperative stay and slightly more postoperative complications. Age alone should not be a contraindication to laparoscopic cholecystectomy in the elderly patient. PMID- 16381356 TI - Minimal-access splenectomy: a viable alternative to laparoscopic splenectomy in massive splenomegaly. AB - BACKGROUND: Laparoscopic splenectomy of normal-sized spleens or in moderate splenomegaly is performed with increasing frequency. By using a modification of the open laparotomy, minimal-access splenectomy is an attractive alternative in severe splenomegaly. METHODS: Between September 2002 and October 2003, 9 patients (mean age, 58.8 years; range, 41 to 72) with severe splenomegaly (mean length, 27.9 cm; range, 23 to 32) underwent minimal-access splenectomy. Indications for splenectomy were non-Hodgkin's lymphoma in 5 cases and idiopathic myelofibrosis in 4. RESULTS: Minimal-access splenectomy was successfully completed in all patients. Mean operative time was 124 minutes (range, 75 to 165). Postoperative complications occurred in 2 cases; one perioperative death occurred in a patient with idiopathic myelofibrosis as a consequence of a secondary blast crisis. Median postoperative hospital stay was 9.1 days (range, 6 to 15). CONCLUSIONS: Minimal-access splenectomy seems to be a viable alternative to laparoscopic splenectomy in cases of severe splenomegaly. It combines the advantages of hand assistance like shorter operative times and increased safety of the procedure to the classical benefits of minimally invasive surgery. PMID- 16381357 TI - Laparoscopic splenectomy reduces the need for platelet transfusion in patients with idiopathic thrombocytopenic purpura. AB - OBJECTIVES: Laparoscopic splenectomy has been increasingly used in patients with idiopathic thrombocytopenic purpura. Because it is associated with minimal abdominal trauma, platelet consumption could be reduced with the laparoscopic approach. The aim of this study was to analyze intraoperative bleeding and the need for apheresis platelets, comparing laparoscopic with open splenectomy. METHODS: Records of 40 patients who underwent splenectomy (20 through laparoscopy and 20 through open surgery) for idiopathic thrombocytopenic purpura were retrospectively reviewed. Intraoperative bleeding and need of perioperative apheresis platelets were evaluated in both groups. Statistical evaluation was conducted using the Mann-Whitney rank test, and differences were considered significant at P<0.01. RESULTS: The mean amount of intraoperative bleeding was less in the laparoscopic group (P<0.01). Apheresis platelets were necessary in all patients in the open group (2 units transfused in 55% and 1 unit in 45% of cases) and only in 30% of cases in the laparoscopic group (1 unit transfused in each case). CONCLUSIONS: Laparoscopic splenectomy is a safe procedure also in patients at high risk for bleeding diathesis. In idiopathic thrombocytopenic purpura, laparoscopic splenectomy should be the gold-standard surgical treatment. Need of platelet transfusion is probably reduced when laparoscopic splenectomy is compared with open surgery in these patients. PMID- 16381358 TI - A "one-stage" laparoscopic procedure for treating choledocholithiasis. AB - OBJECTIVES: A minimally invasive approach is considered the treatment of choice for gallbladder stones. We report our experience with the treatment of choledocholithiasis. METHODS: From January 1993 to December 2002, 3118 patients underwent minimally invasive surgery for symptomatic gallstones, 2681 for gallbladder stones and 437 (14%) for cholecysto-choledocholithiasis. RESULTS: We performed endoscopic retrograde cholangiopancreatography and endoscopic sphincterotomy in 71 patients (18.7%) with high operative risks, transcystic clearance and transcystic drainage in 96 cases (26.2%) and transcholedochal clearance with a T-tube in 270 cases (73.8%). In 2 patients, residual stones were removed with endoscopic retrograde cholangiopancreatography and endoscopic sphincterotomy. Postoperative stay ranged from 4 days to 12 days. No morbidity or mortality occurred. CONCLUSION: In our experience, "one-stage" laparoscopic procedure for cholecystocholedocholithiasis is safe and effective in skilled hands. PMID- 16381359 TI - Treatment of urachal anomalies: a minimally invasive surgery technique. AB - BACKGROUND: Urachal disease is uncommon. The surgical treatment consists of the resection of the urachus throughout its entire length. Our objective is to demonstrate the use of minimally invasive surgery to treat this disease. METHODS: Six patients were studied and diagnosed. The technique used three 10-mm ports on the right hemi abdomen, through which the dissection of the urachus was performed from the umbilical extreme to the bladder. We evaluated the perioperative records to assess morbidity and outcome. RESULTS: Most patients suffered from episodes of umbilical discharge. The diagnosis was made mainly through clinical history and confirmed during the laparoscopic procedure. The urachus was resected throughout its entire length, and we did not perform a segmentary bladder resection in any patient. The average operative time was 66 minutes (range, 42 to 123), and no operative complications were associated with the technique. DISCUSSION: Minimally invasive surgery is a safe and effective procedure that allows the dissection of the urachus through its entire length, providing optimal postoperative results. PMID- 16381360 TI - Chronic pelvic pain: the occurrence of interstitial cystitis in a gynecological population. AB - OBJECTIVE: The objective of this study was to determine what relationship exists between interstitial cystitis and chronic pelvic pain in patients. METHODS: A prospective study of 35 women with a complaint of chronic pelvic pain was performed between August 2002 and September 2003. These patients underwent a workup to exclude other causes of pelvic pain and underwent a laparoscopy and a cystoscopy with hydrodistention at 80 cm of hydrostatic water pressure. Results were obtained and quantified. RESULTS: Twenty-eight patients (80%) were diagnosed with interstitial cystitis, 28 were diagnosed with endometriosis (80%), 24 had both disease entities simultaneously (69%), and 32 (91%) had endometriosis, interstitial cystitis, or both. Three patients (9%) had neither and were diagnosed with other pathologies. CONCLUSIONS: Chronic pelvic pain is a major concern for many women in the United States. Patients with chronic pelvic pain have traditionally been difficult to manage. A large percentage of women presenting with chronic pelvic pain have been shown to have endometriosis, interstitial cystitis, or both. Therefore, an appropriate workup for those individuals with chronic pelvic pain involves not only obtaining a good history and performing a good physical examination, but the possibility of a cystoscopy being performed when a laparoscopy has been deemed necessary for diagnosis of the pain. These procedures can serve as both a means for diagnosis and short-term treatment of these problems when encountered. PMID- 16381361 TI - Microlaparoscopy and a GnRH agonist: a combined minimally invasive approach for the diagnosis and treatment of occlusive salpingitis isthmica nodosa associated with endometriosis. AB - OBJECTIVE: To evaluate whether occlusive salpingitis isthmica nodosa associated with endometriosis can be diagnosed by microlaparoscopy and managed with medical therapy using leuprolide acetate. METHODS: This was a prospective, nonrandomized study conducted at a university hospital and a private community hospital. It included women with occlusive salpingitis isthmica nodosa associated with endometriosis. Diagnosis of salpingitis isthmica nodosa was made via microlaparoscopy with chromotubation. Patients with occlusive salpingitis isthmica nodosa were treated with leuprolide acetate 3.75 mg administered monthly for 6 months. RESULTS: Tubal patency in occlusive salpingitis isthmica nodosa following medical therapy with leuprolide acetate was evaluated. Thirteen of 16 (81.3%) women with bilateral salpingitis isthmica nodosa achieved patency of both fallopian tubes following treatment with leuprolide acetate; 3 of 16 (18.8%) developed patency in one of the fallopian tubes. All 5 women with unilateral SIN demonstrated bilateral patency following medical therapy. CONCLUSION: Diagnosis of occlusive salpingitis isthmica nodosa can be made by microlaparoscopy. These preliminary results suggest that medical therapy with leuprolide acetate may be the first-line treatment modality for women with occlusive salpingitis isthmica nodosa associated with endometriosis, possibly avoiding a more invasive surgical procedure. PMID- 16381362 TI - Laparoscopic myomectomy for very large myomas using an isobaric (gasless) technique. AB - OBJECTIVES: Laparoscopic myomectomy using pneumoperitoneum for large myomas (> or = 8 cm) is hindered by several factors, such as the increased operative time, the risk of perioperative bleeding, and the risk of conversion to laparotomy. With the introduction of isobaric laparoscopy using abdominal wall lifting, this procedure can be performed using conventional surgical instruments introduced through small abdominal incisions. The aim of this study was to evaluate the feasibility, reproducibility, and safety of isobaric laparoscopic myomectomy for very large myomas > or = 10 cm using a subcutaneous abdominal wall-lifting device. METHODS: A series of 24 consecutive patients with at least 1 symptomatic myoma > or = 10 cm underwent a gasless laparoscopic myomectomy with the Laparotenser device. Conventional long laparotomy instruments were used. RESULTS: Gasless laparoscopic myomectomy was successful in all 24 consecutive patients. The size of the dominant myoma varied from 10 cm to 20 cm. The median operating time was 93 minutes. The median postoperative drop in hemoglobin was 2.8 g/dL. No surgical complications occurred. The median hospital stay was 2.8 days. CONCLUSION: Gasless laparoscopic myomectomy is feasible, reproducible, and safe for removing very large myomas. Therefore, it can represent an excellent option for the minimally invasive removal of very large myomas. PMID- 16381363 TI - Comparison of 5-, 10-, and 15-point laparoscopic ovarian electrocauterization in patients with polycystic ovarian disease: a prospective, randomized study. AB - OBJECTIVE: We compared 12-month pregnancy and live birth rates in patients with polycystic ovarian disease undergoing 5-, 10-, and 15-point laparoscopic ovarian electrocauterization. METHODS: This was a prospective, randomized study performed at the Dabirashrafi Fertility and Endoscopy Research Center, Tehran, Iran. The study included 187 patients with polycystic ovarian disease who were randomly assigned to 3 groups. Group I comprised 67 patients whose ovaries received 5 point electrocauterization. Group II comprised 57 patients whose ovaries received 10-point electrocauterization. Group III comprised 63 patients whose ovaries received 15-point electocauterization. Laparoscopic ovarian electrocauterization with a unipolar current was used. The main outcome measures were 12-month pregnancy and live birth rates. RESULTS: Patients were homogeneous for age, body mass index, and type and duration of infertility. Twenty pregnancies resulted in Group I, with a pregnancy rate of 29.9% (20/67) and a live birth rate of 20.9% (14/57). Eighteen pregnancies resulted in Group II, with a pregnancy rate of 31.6% (18/57), and a live birth rate of 28.1% (16/57). Thirty-three pregnancies resulted in group III, with a pregnancy rate of 52.4% (33/63), and a live birth rate of 47.6% (30/63). Comparison of Group III with Groups I and II revealed a statistically significant increase in pregnancies (P=0.016) and live birth rates (P=0.004). CONCLUSION: We recommend 15-point electrocauterization of ovaries in patients with polycystic ovarian disease. PMID- 16381364 TI - Laparoscopy or laparotomy for the management of endometrial cancer. AB - OBJECTIVE: The aim of this study was to evaluate the feasibility of laparoscopy in the management of early stage endometrial cancer. METHODS: Fifty-two patients with endometrial cancer who underwent surgical staging consisting of total hysterectomy, bilateral salpingo-oophorectomy with pelvic lymph node dissection, and cytology between 1998 to 2002 were included in the study. Laparotomy and laparoscopy were randomly offered to patients upon admittance. RESULTS: Of 52 patients, 26 underwent laparotomy and the remaining 26 underwent laparoscopic staging surgery. No significant difference existed between the demographic characteristics of the 2 groups. The mean number of harvested lymph nodes was 18.2 in the laparoscopic group and 21.1 in the laparotomic group (P>0.05). Pelvic lymph node metastases were detected in 7.7% of the patients in the laparoscopy group and 15.4% in the laparotomy group, and the difference was not significant. Adjuvant radiotherapy was applied later to 42.3% of the laparoscopy group and 38.5% of the laparotomy group. Operative morbidity was higher in the laparotomy group mainly because of postoperative wound infection, and the patients in the laparotomy group had a longer hospital stay. CONCLUSION: Laparoscopic surgery is a method that can be applied as well as laparotomy in the management of endometrial cancer. Lymph node number and detection of lymph node metastasis did not differ significantly in laparotomic and laparoscopic approaches. Wound infections were more frequent in laparotomies. PMID- 16381365 TI - Laparoscopic myomectomy with lateral dissection of the uterine artery. AB - BACKGROUND: We assessed the results and impact of lateral uterine artery dissection on clinical outcome following laparoscopic myomectomy. METHODS: We retrospectively analyzed the clinical data for 27 laparoscopic myomectomy cases (Group I) and 54 laparoscopic myomectomy cases combined with lateral uterine artery dissection (Group II) between January 2001 and August 2004 in one center. Only 81 patients who had dominant fibroids between 4 cm and 10 cm in diameter were included in the study. We assessed the clinical outcomes: perioperative blood loss, operating time, hospital stay, complications, hemoglobin decrease, inflammatory response, and tissue markers (C-reactive protein, white blood cells, creatinine kinase) changes. RESULTS: The mean operating time was 70.37 minutes in group I and 78.61 minutes in group II. The mean length of hospital stay was 2.7 days versus 2.2 days, respectively (P>0.05). The difference in intraoperative blood loss was 70.1 mL (147.7 mL vs 77.3 mL, Group I) and 33.9 mL (105 mL vs 71.1 mL, Group II); estimated postoperative blood loss was statistically significant (P<0.001, P<0.05, respectively). Group 2 demonstrated a less intense stress response in C-reactive protein (P<0.001) and white blood cell count (P<0.05). CONCLUSION: The dissection of the uterine artery in laparoscopic myomectomy is a feasible operative procedure with a low rate of complications. The procedure reduced perioperative blood loss and resulted in significant improvement in fibroid-related symptoms. PMID- 16381366 TI - Safety and efficacy of metallic stents in the management of colorectal obstruction. AB - BACKGROUND: The use of self-expandable metallic stents in the management of obstructing colorectal cancer has been described with increasing frequency in the literature. Our goal was to evaluate the efficacy and associated morbidity of the use of self-expandable metallic stents to relieve colorectal obstruction at our institution. METHODS: A retrospective chart review of patients who underwent colorectal stent placement between December 2001 and December 2003 in a tertiary referral center was performed. RESULTS: Stents were placed successfully in 17 of 21 patients (81%) with colorectal obstruction. Placement was achieved endoscopically in 13 patients and radiologically in 4. Ten self-expandable metallic stents were used as a bridge to surgery, and 7 were used for palliation. The obstructions were located in the sigmoid colon (11 patients), the rectosigmoid (3), the splenic flexure, the hepatic flexure, and the rectum. Malignant obstruction was noted in 14 patients. One patient with malignancy experienced a sigmoid perforation, and 2 patients with benign disease had complications (1 stent migration and 1 re-obstruction). Stent patency in obstruction secondary to colonic adenocarcinoma was 100% in our follow-up period (range, 5 to 15 months). CONCLUSIONS: The use of stents as a bridge to surgery is associated with low morbidity, allows for bowel preparation, and thus avoids the need for a temporary colostomy. Long-term patency suggests that stents may allow for the avoidance of an operation in patients with metastatic disease and further defines their role in the palliation of malignant obstruction. Further prospective randomized studies are necessary to fully elucidate the use of stents in the management of colorectal cancer. PMID- 16381367 TI - A training module for laparoscopic urology. AB - OBJECTIVES: A fellowship training model in laparoscopic urological surgery has been established for interested urologists to help them proceed from the pelvic trainer/animal laboratory environment to safe clinical practice. The objective of the model is to provide trainees with clinical experience under direct mentor supervision before embarking on independent laparoscopic urological surgery at their own base hospitals. METHODS: The fellowship model incorporates 9 fluid phases: Phase 1 to complete basic and advanced training courses. Phase 2 to practice at home or in the office using pelvic trainers. Phase 3 to proceed to an animal laboratory course. Phase 4 to visit centers of international repute to observe high-volume laparoscopic urology. Phase 5 to observe the mentor perform several major renal laparoscopic cases. Phase 6 to perform several hand-assisted renal procedures under direct mentor guidance at the mentor hospital. Phase 7 to perform several laparoscopic or retroperitoneoscopic renal procedures, or both, under direct mentor guidance at the mentor hospital. Phase 8 to mentor assisted trainees to start laparoscopic surgery at their own hospitals. Phase 9 to practice laparoscopic urology independently. RESULTS: So far, 9 trainees have participated in the fellowship. Six have reached phase 9 with independent practice, 2 others are in phase 8, and 1 is in phase 7. Skills development has been steady, with progressive acquisition of surgical dexterity and spatial orientation. CONCLUSION: This fluid fellowship model provides urologists with clinically applicable teaching experience to learn a relatively new surgical concept safely and effectively, thereby promoting clinical governance. It may be possible for other centers to establish similar fluid "mini" fellowships to help disseminate laparoscopic surgical skills. PMID- 16381368 TI - Heated, humidified CO2 gas is unsatisfactory for awake laparoscopy. AB - BACKGROUND: The necessity for general anesthesia represents an impediment to using a laparoscopic approach for some procedures that are otherwise performed with the patient under local anesthesia using a conventional open technique. Heating and humidifying the insufflation gas reportedly reduces perioperative pain associated with a CO2 pneumoperitoneum, thus enabling awake laparoscopy. METHODS: Two cases are reported herein of laparoscopy performed with the patient under local anesthesia using heated, humidified CO2 gas for the pneumoperitoneum. RESULTS: Both patients experienced pain with insufflation of heated, humidified CO2 gas of sufficient magnitude that the procedure could not be performed. The CO2 gas was washed out and replaced with helium gas insufflation with complete resolution of pain. The laparoscopic procedures were accomplished without further discomfort with local anesthesia and using a helium gas pneumoperitoneum. CONCLUSIONS: Heated, humidified CO2 gas insufflation does not reduce pain sufficiently to permit satisfactory performance of laparoscopy with local anesthesia, especially when full volume insufflation is required. Cold, dry helium gas produces no pain. The theory that cold, dry insufflation gas is a source of peritoneal pain during laparoscopy needs to be reassessed. PMID- 16381369 TI - Metastatic spread to a percutaneous gastrostomy site from head and neck cancer: case report and literature review. AB - BACKGROUND: The placement of percutaneous endoscopic gastrostomy tubes is a common procedure in patients with head and neck cancer who require adequate nutrition because of the inability to swallow before or after surgery and adjuvant therapies. A potential complication of percutaneous endoscopic gastrostomy tubes is the metastatic spread from the original head and neck tumor to the gastrostomy site. METHODS: This is a case of a 59-year-old male with a (T4N2M0) Stage IV squamous cell carcinoma of the oropharynx who underwent percutaneous endoscopic gastrostomy tube placement at the time of his surgery and shortly thereafter developed metastatic spread to the gastrostomy site. A review of the published literature regarding the subject will be made. RESULTS: Twenty nine cases of percutaneous endoscopic gastrostomy site metastasis occurring in patients with head and neck cancer have been previously reported in the literature. The pull-through method of gastrostomy tube placement had been used in our patient as well as in the majority of the other cases reviewed in the literature. CONCLUSION: The metastatic spread of head and neck cancer to the percutaneous endoscopic gastrostomy site is a very rare occurrence. The direct implantation of tumor through instrumentation is the most likely explanation for metastasis; however, hematogenous seeding is also a possibility. To prevent this rare complication, other techniques of tube insertion need to be considered. PMID- 16381370 TI - Laparoscopic treatment of Bouveret's syndrome presenting as acute pancreatitis. AB - Gastric outlet obstruction as a result of gallstone (Bouveret's syndrome) is a rare but serious complication of cholelithiasis. Although patients present with persistent vomiting, colicky epigastric pain and dehydration, the clinical features of the Bouveret's syndrome are not pathognomonic. Due to its rarity, the diagnosis and treatment represent a challenge for the surgeon. In most of the reported cases, the diagnosis was made at the time of laparotomy. We report an unusual clinical presentation of Bouveret's syndrome with mild acute pancreatitis that was treated laparoscopically. To our knowledge, this is the first described case. Cause, clinical presentation, methods of diagnosis, and options for management of Bouveret's syndrome are also discussed. PMID- 16381371 TI - Exacerbation of occult femoral hernia during laparoscopic prostatectomy. AB - Laparoscopic prostatectomy has been accepted as an appropriate treatment for prostate cancer because of the shorter hospital stay and quicker recovery. We present a rare complication of groin hernia with incarceration and necrosis of small bowel following laparoscopic prostatectomy. Occult hernias and small fascia defects may not always be apparent pre-operatively, but extension of pneumoperitoneal insufflation to extraperitoneal compartments should alert the surgeon to the possible presence of such a defect. PMID- 16381372 TI - A case of emphysematous cholecystitis managed by laparoscopic surgery. AB - BACKGROUND: Emphysematous cholecystitis is a rare condition caused by ischemia of the gallbladder wall with secondary gas-producing bacterial proliferation. The pathophysiology and epidemiology of this condition differ from that in gallstone related acute cholecystitis. This report illustrates a case of emphysematous cholecystitis successfully treated by laparoscopic surgery. METHODS: An 83-year old female patient was admitted to the hospital with acute abdominal syndrome. Clinical examination and blood tests suggested acute cholecystitis. Plain radiography revealed a circular gas pattern in the right upper quadrant suggestive of emphysematous cholecystitis. Subsequent computed tomography confirmed the presence of gas in the gallbladder wall and a gas-fluid level within the organ. RESULTS: Emergency laparoscopic cholecystectomy was successfully performed during which bubbling of the gallbladder wall was observed. Intraoperative cholangiography revealed no bile duct stones or biliary obstruction. The patient made an unremarkable recovery from surgery with no postoperative complications or admission to the intensive care unit. Pathological analysis revealed full-thickness infarctive necrosis of the gallbladder. Bacterial cultures grew Clostridium perfringens. CONCLUSIONS: This case illustrates a typical case of emphysematous cholecystitis successfully treated by laparoscopic surgery. It contributes to suggestions from other reports that this condition can be safely treated by the laparoscopic approach. PMID- 16381373 TI - Laparoscopic cholecystectomy in a patient with a biventricular cardiac assist device. AB - Evaluation and management of abdominal pathology in patients with ventricular assist devices is likely to become increasingly important as the utilization of these devices expands. Ventricular assist devices represent a class of intracorporeal or paracorporeal mechanical devices that augment cardiac output in patients with congestive heart failure. Patients with ventricular assist devices supporting both right and left ventricles (biventricular assist devices) are uniquely challenging to the general surgeon because these devices restrict direct access to the abdominal cavity and because of the perioperative implications of biventricular heart failure. We describe herein the first reported successful laparoscopic cholecystectomy in a patient with a paracorporeal biventricular assist device. Cholecystectomy was performed in this patient for acute cholecystitis that occurred while the patient was awaiting heart transplantation. Our results add weight to the small body of evidence that laparoscopy is well tolerated in ventricular assist devices patients. The unique aspects of the biventricular assist device patient make laparoscopic abdominal intervention particularly suitable in this patient population. PMID- 16381374 TI - Laparoscopic management of gallbladder-associated ectopic liver. AB - Ectopic liver is a rare entity discussed infrequently in the surgical literature. Liver ectopia develops due to rests of liver parenchyma retained at various intraperitoneal or intrathoracic locations during migration of the liver proper during embryologic development. It is usually found during exploration for other diagnoses, commonly diseases of the biliary tree. We report a case in which a 3.7 cm mass associated with the fundus of the gallbladder was visualized preoperatively by computed tomography and identified histologically as ectopic liver after diagnostic laparoscopy and cholecystectomy with en bloc resection of the associated mass. PMID- 16381375 TI - Colonoscopic diagnosis of appendiceal intussusception: case report and review of the literature. AB - Intussusception of the appendix is an extremely rare condition. Although approximately 200 cases of appendiceal intussusception have been reported in the literature, very few have ever been diagnosed preoperatively. We report a case of appendiceal intussusception secondary to endometriosis in an otherwise healthy female. The case was diagnosed preoperatively by colonoscopy and treated surgically at laparoscopy. We review the literature of appendiceal intussusception and discuss the associated conditions, diagnosis, and a classification scheme for this unusual finding. PMID- 16381376 TI - Laparoscopic abdominal cerclage. AB - Abdominal cerclages are necessary when the standard transvaginal cerclages fail or anatomical abnormalities preclude the vaginal placement. The disadvantage of the transabdominal approach is that it requires at least 2 laparotomies with significant morbidity and hospital stays. We discuss a case of abdominal cerclage performed laparoscopically. We feel it offers less morbidity and in the proper hands eliminates or significantly shortens hospital stays. PMID- 16381377 TI - Torsion of the greater omentum: preoperative computed tomographic diagnosis and therapeutic laparoscopy. AB - Primary or idiopathic segmental infarction of the greater omentum is a rare surgical condition. We describe a case of omental torsion in an adult patient who was diagnosed preoperatively by contrast-enhanced computed tomography and managed by laparoscopy. PMID- 16381378 TI - Emergency laparoscopic unroofing of a ruptured hepatic cyst. AB - The use of laparoscopic techniques in the emergency treatment of liver cyst complications has not been reported in the literature. Laparoscopic surgery is a well-established treatment modality for giant hepatic cyst in the elective setting. We report a case of laparoscopic unroofing of a 15-cm ruptured hepatic cyst, which had recurred after previous percutaneous alcohol instillation therapy. Laparoscopic ultrasound was used to delineate the extent of the cyst while a Harmonic scalpel was used for dissection. The patient made an uneventful recovery and was discharged 4 days after the operation. PMID- 16381379 TI - Processing complaints and disciplining nurses. An overview of the investigative process by the Kansas State Board of Nursing. AB - This is a brief review of the process the Kansas State Board of Nursing would utilize when investigating a complaint and determining discipline against a licensee, and does not include all points of law that could apply. Although the Kansas Administrative Procedures Act was discussed, there are many sections of the statute that were not addressed. The Board of Nursing has published two pamphlets "Your Rights Before the Kansas State Board of Nursing" and "The Investigative Process" which are valuable to the nurse in understanding the process. These are available on the KSBN's website www.ksbn.org or by mail. PMID- 16381380 TI - Emergency care EMTALA. Implementation and enforcement issues. PMID- 16381381 TI - Standards for privacy of individually identifiable health information. HIPAA implementation. PMID- 16381382 TI - [Brain of patients with chronic schizophrenia and neurotransmitters]. PMID- 16381383 TI - [Relationship between the courses of clinical Features of patients with schizophrenia in adolescents and admission to psychiatric clinic]. AB - In order to improve diagnosis of schizophrenia with onset in adolescents at an early stage, we investigated in detail the clinical features of 74 patients with schizophrenia, (23 males) at adolescents psychiatric clinic. Many of the subjects had been suffering from the illness about 14 years old but had not undergone their first psychiatric examination until a few years later. A high percentage (more than 80%) of our subjects presented psychiatric symptoms such as delusional remembrance, delusional moods, delusions of persecution and hypobulia. Additionally, more than 60% of our subjects presented auditory hallucinations. In general, teenage patients with schizophrenia onset show vague symptoms such as anxiety, embarrassment and strange moods rather than obvious hallucinations. Nevertheless, it was possible to identify certain clinical features of this disorder in adolescents: many patients suffer delusional remembrance, delusional moods and delusions of persecution immediately after the onset of the illness. Gradually, problematic behaviors such as anorexia, self injury, offences against their families, voluntary vomiting, etc., develop, but patients do not always receive psychiatric examination at this stage. After socially obvious problems such as school refusal, withdrawal from social activities and lowering of school record develop over a period of time, patients may be urged to undergo psychiatric examination. Our research again underlines the difficulty of achieving diagnosis of schizophrenia at an early stage. The key to early diagnosis appears to be the accurate identification of psychiatric symptoms in the early stages of the illness at school, or at home if possible, before socially problematic behaviors arise. PMID- 16381384 TI - [Remarkably low blood pressure induced by combination of SSRI and a small dose of TCA]. AB - Recently, selective serotonin reuptake inhibitors (SSRI) have been frequently used as the first choice of antidepressants because of their safety in comparison with tricyclic antidepressive agents (TCA). On the other hand, when SSRIs are not effective, they are frequently replaced with another antidepressant. In that case, a small dose of a TCA is often added without cross tapering. We report two elderly patients with depression who exhibited markedly low blood pressure when a small dose of a TCA was added to a SSRI. The combination of a SSRI and a TCA can be hazardous because it can cause unexpected adverse effects especially in elderly patients. PMID- 16381385 TI - [Realities and function of 50-bed stress care ward in the non-profit private psychiatric hospital]. PMID- 16381386 TI - [Role of the stress care ward in the psychiatric hospital: Report on hospitalized treatment for patients with depressive disorder]. PMID- 16381387 TI - [Why does hospitalized care for patients with depressive disorder take place in the stress care ward?]. PMID- 16381388 TI - [Meaning of rest in therapy for the depressive disorders-- role of the stress care ward]. AB - The Meaning of Rest in Therapy for the Depressive Disorders-The Role of Stress Care Ward Rest is necessary in the therapy for the depressive disorders, even in the case that pharmcotherapy or psychological treatment seems to be effective. In this paper, the meaning of rest is examined, which is inclined to be underestimated. Especially, the meaning of admission in therapy for the depressive disorders is recognized and re-estimated. Rest is minimum environmental requirement for restoring spontaneous healing, and also is maximum therapeutic strategy for activating it. Systematic medical provision of rest is admission to stress-care ward, and it is given as environment in which stress related sickness patients are able to be really rest. According to the study at the stress-care ward in the Siranui hospital in 2004, standard citizens were admitted with moderate depressive state, and around 90% patients were recovered after about three month therapy. Most important important therapeutic factors were supposed to be abundant stuff supply and team therapy. There are not enough hospital beds for the depressive disordered patients as adequate environment for rest. We hope radical medical policy change in this area in near future. We think that evidence based study is needed for examining effectiveness of rest and admission to stress-care ward as therapeutic strategy. PMID- 16381389 TI - [Operative management of the stress care unit in the psychiatric hospital]. PMID- 16381390 TI - [Suicide prevention for patients with depressive disorder in Japan]. PMID- 16381391 TI - [Early-stage schizophrenia in cases of committed suicide]. PMID- 16381392 TI - [Treatment and prevention for suicidal patients with severe personality disorders]. AB - Suicide is a complex behavior of multiple, inter-related factors. The author, in this paper, discusses several issues of the psychodynamic psychotherapy for chronic suicidal patients with severe personality disorders. With regard to psychotherapy for them, the author firstly points out the treatment difficulties that stem from the training system both of psychiatry and psychotherapy where students are not systemically trained with treatment of suicidal behavior. Secondly he discusses psychodynamic evaluation, therapeutic attitude, treatment setting, counter-transference, and grief work. In terms of psychodynamic evaluation, it is suggested that a therapist needs to recognize self-maintaining function, unconscious fantasy of re-union with great pleasure, internal object relations that lead to self-destructiveness, resentment toward self, capacity for loving good aspects of personality, specific sensitiveness to certain object loss, disorganization of self representation, and ego regression. With regard to, the attitude of, the therapist and the treatment setting, the author stresses that psychotherapy should be conducted in a highly structured treatment setting with the attitude of the therapist such as active intervention, honesty (openness), awareness of treatability, and conviction. Countertransferences frequently develop based on the feelings that the therapist experiences narcissistic injury with regard to his/her own ego ideal in the psychotherapy process, in which he/she develops the various defense mechanism against the increasing aggression of his/her own. A therapist needs to be aware of the patient's dependency, make frequent interpretations of patient's destructiveness, and make effective use of the consultation at the time when he/she places himself/herself at the impasse of treatment. Finally, the author emphasizes the grief work for both therapist and family in the case when the patient unfortunately has committed suicide and open communications among therapist, other staff involved and family members should be strongly recommended. These works not only help persons concerned to work through their mourning work, but also prevent the transmission of suicides over generations. PMID- 16381393 TI - [Suicide in children and the prevention]. AB - There have been studied on suicide in children and adolescents in Western countries, literature review was carried out for the forth coming needs of protective intervention for suicidal children in Japan. Suicide is very rare in children with gradual increase towards adolescents. Suicide in children has association with family history, psychiatric disorders of both children and parents, especially depression and addictive disorders, cognitive and emotional disturbance in children, demographic characteristics, family discord, family psychopathology and child abuse, accessibility of lethal suicide methods. Identification of risk factors as foci for intervention is important for suicide prevention. Although there are some programs for prevention, their theoretical bases have been hardly presented and their efficacy are not acceptable. Future evaluative research is essential, and at moment, network of medical, educational and child welfare professionals is important to support children and family at risk where there is much to do with psychiatry to take role. PMID- 16381394 TI - [Prevention of suicide in the elderly]. AB - In order to establish a comprehensive community intervention program for preventing suicide in late life, a series of observation and intervention studies was conducted in an urban residential district where the elderly population was rapidly increased. In cross-sectional studies, impaired instrumental activities of daily living, lack of social support, and depressive symptoms were independently associated with suicidal ideation in elderly community residents. In addition, mental disorders, particularly depressive and alcohol-related disorders, were found to be strongly associated with recent suicidal ideation. In prospective cohort studies, low motor function or severe pain, alcohol problems, and depressive symptoms independently predicted male suicidal ideation, whereas only depressive symptoms predicted female suicidal ideation. In intervention studies, education, consultation, increased community activity, and screening programs decreased the proportion of residents who indicated a lack of social support, and reduced depressive symptoms (GDS). Additionally, outreach and case management programs improved mental-health wellbeing (WHO-5), and reduced suicidal ideation among elderly with depressive disorders. Screening for depressive symptoms followed by diagnostic evaluation of mental disorders may provide a practical and effective strategy for case identification. However, it should be noted that physical distress and alcohol problems might increase suicide risk irrespective of depression, especially among elderly males. The combination of population and high-risk strategies is recommended for a comprehensive community intervention program for preventing elderly suicide in urban residential districts. PMID- 16381395 TI - [Clinical cognitive therapy]. AB - Cognitive therapy was originally developed as a short-term approach to the treatment of depression. In recent years, it has been applied, either alone or in combination with pharmacotherapy, to a wide range of mental disorders, from anxiety, eating, and substance use disorders to personality, bipolar, and schizophrenic disorders. In this article, how to use cognitive therapy for treating psychiatric outpatients will be illustrated. Also, clinical decision analysis, a quantitative method for synthesizing the best research evidence and the patient's preference, will be highlighted in the acute and maintenance treatment of recurrent depression. PMID- 16381396 TI - [Occupational mental health services and mental disorders]. PMID- 16381397 TI - What have we done for you lately? Liability reform efforts looking positive. PMID- 16381398 TI - Big help for small hearts. PMID- 16381399 TI - Katrina took, you gave. Arkansas health care providers gave selflessly in the wake of tragedy. PMID- 16381400 TI - Could HIT transform health care? Hospitals, health care providers hold the key. PMID- 16381401 TI - The obstetrical dilemma. PMID- 16381402 TI - A success story with chronic hepatitis C. PMID- 16381403 TI - Meet our members: Jim Mark Ingram, MD. PMID- 16381404 TI - Sacroplasty. PMID- 16381405 TI - Pharmacist activity in the United States: view of a Japanese government regulator and pharmacist. PMID- 16381406 TI - Discovering diabetes. PMID- 16381407 TI - Ready or not, here comes medication therapy management! PMID- 16381408 TI - Pharmacy's dedication in meeting hurricane evacuees' needs. PMID- 16381409 TI - Hurricane Katrina: pharmacists making a difference. PMID- 16381410 TI - Caring for animal patients following Hurricane Katrina. PMID- 16381411 TI - American Pharmacists Association Foundation: a catalyst for change. PMID- 16381412 TI - Impact of Hurricane Ivan on pharmacies in Baldwin County, Alabama. AB - OBJECTIVE: To evaluate the impact of Hurricane Ivan, which made landfall east of Mobile, Alabama, on September 16, 2004, on pharmacies in the affected areas. DESIGN: Retrospective cross-sectional analysis. SETTING: Baldwin County, Alabama. INTERVENTIONS: Pharmacy community rapid-needs-assessment survey. PARTICIPANTS: 41 hospital and community (chain and independent) pharmacies. MAIN OUTCOME MEASURES: Posthurricane pharmacy hours of operations, prescription volumes, infrastructure damage, and prehurricane disaster planning. RESULTS: During the week of the hurricane, both chain and independent community pharmacies within the evacuation zone worked significantly fewer hours (46% and 49%, respectively) and dispensed significantly fewer prescriptions (37% and 52%) compared with the same week of the prior year. Overall, 40% of pharmacies depleted their supplies of certain medications (e.g., anxiolytics, antihypertensives). A total of 60% of the chain and independent pharmacies outside the evacuation zone closed because of loss of electricity, but pharmacies with a generator were significantly less likely to report having turned away patients. The proportion of pharmacies that had a disaster plan but turned away patients or rationed or ran out of medications was similar to that of pharmacies without a disaster plan. CONCLUSION: Although Hurricane Ivan primarily affected the operation of pharmacies within the evacuation zone, pharmacies in the surrounding area were also affected because of loss of power. Emergency management officials should evaluate the efficacy of specific guidelines outlined in disaster plans and identify ways to deliver essential medications to people in disaster-affected areas. PMID- 16381413 TI - Job turnover intentions among Certified Pharmacy Technicians. AB - OBJECTIVE: To describe the job withdrawal intentions of a nationwide sample of Certified Pharmacy Technicians (CPhTs). DESIGN: Cross-sectional descriptive design. SETTING: A random sample of 3,200 CPhTs whose names and contact information were acquired from the Pharmacy Technician Certification Board. INTERVENTIONS: Mailed survey. MAIN OUTCOME MEASURES: Responses to questions eliciting job withdrawal intentions, reasons for remaining/staying with the employer, and various quality of worklife attitudes. Employment intentions were regressed over quality of worklife and practice environment variables in a forward-conditional logistic regression procedure. RESULTS: Nearly 4 in 5 respondents indicated intentions to remain with the current employer during the next 12 months. Relationships with coworkers, good benefits, and work schedule were frequently cited as reasons to stay. Poor salary, lack of advancement opportunity, and insufficient staffing were cited frequently as factors among those indicating intentions to leave. Mean hourly wages differed among respondents with intention to leave due to salary, those not citing salary as a reason to remain or leave, and those citing salary as a reason for staying. Intention to remain was associated with higher pay, perceived employer support, number of years with the current employer, and lower career turnover intentions, with organizational commitment acting as a powerful mediator for all of the variables. CONCLUSION: Differences in CPhTs' future employment intentions were described more comprehensively by perceptions of the employing organization than by perceptions of managers/supervisors and coworkers or personal/work environment variables. Implications for pharmacy organizations are discussed. PMID- 16381414 TI - Assessing risk for loss of rural pharmacy services in Minnesota. AB - OBJECTIVE: To quantify rural communities' risks for losing access to local pharmacy services using a newly developed risk-assessment tool. DESIGN: Cross sectional study. SETTING: Minnesota in June through August 2003. PARTICIPANTS: Pharmacy owners and managers in rural communities with populations less than 5,000 residents and only one community pharmacy. INTERVENTION: Self-administered questionnaire completed by study participants. MAIN OUTCOME MEASURES: Each answer was given a score based on the related factor's ability to affect risk of pharmacy closure. Scores were summed to give a comparative risk score with a positive score indicating higher risk; these scores could range from -13.5 to +30.75, depending on responses. RESULTS: From 126 communities outside of the Minneapolis-St. Paul metropolitan area with populations of fewer than 5,000 residents and only one community pharmacy, 81 (64.3%) usable responses were received. The calculated risk scores ranged from -7 to +12.3, with mean (+/- SD) scores of 1.55 +/- 4.17. Factors that contributed most to rural pharmacies' risk index were dramatic decreases in community populations within 5 years, poor community support for the pharmacy, owner's intent to sell pharmacy within 2 years, less than dollar 750,000 in annual prescription revenue, perceived erosion of total market share of prescriptions, difficulties in recruiting full-time pharmacist-employees, lack of access to local primary care, and perceived likelihood that local primary care clinics would not be active in 5 years. Approximately 26% of pharmacy owners responding expressed interest in selling their pharmacy within 3 years, and 62% were interested in selling within 10 years. CONCLUSION: The tool developed for this study allows prospective assessment of risk, which can facilitate a greater awareness by local and regional officials and consumers of the possibility of loss of rural pharmacies, assist in targeting resources to areas most in need, and serve as a bridge to collaborative relationships aimed at ensuring local access to both pharmacy services and the knowledge of a pharmacist. PMID- 16381415 TI - Student pharmacist perspectives of rural pharmacy practice. AB - OBJECTIVE: To evaluate the level of interest of upper-level doctor of pharmacy students in rural pharmacy practice, rural pharmacy ownership, and a proposed rural pharmacy practice model. DESIGN: Cross-sectional study. SETTING: Minnesota, North Dakota, and South Dakota in October through December 2003. PARTICIPANTS: Third- and fourth-year professional student pharmacists at the University of Minnesota, North Dakota State University, and South Dakota State University. INTERVENTION: Self-administered questionnaire completed by study participants. MAIN OUTCOME MEASURES: Student interest in rural pharmacy practice and potential future directions for maintaining access to medications and the knowledge of pharmacists in rural communities. RESULTS: Of 177 respondents, 62.7% has given serious consideration to practice in rural communities with populations of 5,000 or fewer residents. Of these 111 students, 81% would be interested in the proposed practice model explained in the survey. Of the 108 total students expressing interest in the proposed rural pharmacy practice model, 63% had not previously given serious consideration to pursuing ownership (full or partnership) of a community pharmacy. CONCLUSION: Students from the schools that have traditionally supplied the majority of Minnesota rural pharmacists indicated an interest in practicing in rural areas as long as opportunities align with their personal and professional interests. PMID- 16381416 TI - Effects of empowerment on pharmacists' organizational behaviors. AB - OBJECTIVE: To investigate the effects of power factors, need for achievement, and empowerment on commitment, loyalty, identification, and job turnover intention among pharmacists. DESIGN: Cross-sectional study. SETTING: United States. PARTICIPANTS: 447 licensed pharmacists nationwide. INTERVENTION: Self administered questionnaire. MAIN OUTCOME MEASURES: Structural equation modeling was used to assess the fit of the theoretical model and examine the effects of empowerment on pharmacists' behaviors within their organizations using pharmacists' self-reports. RESULTS: An overall response rate of 42.2% was obtained. The test of the hypothesized model using structural equation modeling resulted in a satisfactory fit. The effects of power factors and need for achievement on psychological empowerment (gamma11 = .75, gamma12 = .27) and structural empowerment (gamma21 = .81, gamma22 = .20) were examined. Also, the effects of psychological empowerment and structural empowerment on loyalty (beta31 = .05, beta32 = .69), commitment (beta41 = - .09, beta42 = .92), and identification (beta51 = .05, beta52 = .78) were analyzed. Finally, the effects of loyalty (beta63 = -.24), commitment (beta64 = -.74), and identification (beta65 = .35) on job turnover intention were assessed. CONCLUSION: Kanter's theory, which maintains that structures within organizations have an impact on organizational behaviors, was supported by our findings. Pharmacists' organizational behaviors such as commitment, loyalty, identification, and job turnover intention are influenced by structural empowerment. Given the pharmacist supply-and-demand imbalances of the past few years, organizations should make every effort to retain the pharmacists currently in their employ. PMID- 16381417 TI - Lifestyle modification counseling of patients with dyslipidemias by pharmacists and other health professionals. AB - OBJECTIVES: To establish whether patients who are taking lipid-lowering medications receive information on lifestyle modifications from health care providers when originally prescribed and whether they continue to receive follow up information on lifestyle modifications, and to establish where patients with dyslipidemias are receiving information about lowering their serum cholesterol levels through lifestyle modifications. DESIGN: Cross-sectional survey. SETTING: Two community pharmacies and two hospitals in two medium-sized cities in the midwestern area of the United States. PARTICIPANTS: 234 patients taking medication to lower serum lipids. INTERVENTION: Paper-based survey. MAIN OUTCOME MEASURE: Responses to survey items. RESULTS: Nearly three quarters (73.9%) of participants received information about lowering their serum lipids through lifestyle modifications when they were first diagnosed with elevated serum cholesterol concentrations. Of these, most (83.8%) said that the information came from their physician. Fewer than one half (48.3%) of all participants said that they continued to receive this type of information. Those who received lifestyle modification information at their original diagnosis and who continued to receive this type of information were more likely to be actively trying to lower their serum lipid levels through diet (93.1%) and exercise (71.6%). Participants visited their pharmacy more often than their physician's office each year, yet they recalled pharmacists offering less patient counseling on lifestyle modifications than did physicians and nurses. CONCLUSION: Despite being well positioned to assist patients with elevated serum cholesterol concentrations, pharmacists offer less patient counseling about therapeutic lifestyle modifications compared with physicians and nurses. PMID- 16381418 TI - Medication use patterns and health outcomes among patients using a subsidized prescription drug program. AB - OBJECTIVE: To evaluate medication adherence, medication safety, health care utilization, and health outcomes among patients enrolled in a subsidized prescription program. DESIGN: Cross-sectional study. SETTING: Conducted as part of the Prescription Access program, a subsidized prescription program serving indigent patients residing in Franklin County, Ohio. PATIENTS: Patients qualifying for enrollment in the program were uninsured and had a household income of 200% or less of federal poverty level. Approximately 5% of the 2,500 patients (mean age, 70.6 years) enrolled in the program were systematically selected from a computer-generated patient enrollment report. INTERVENTION: Telephone interviews conducted by a pharmacist or advanced student pharmacist between January and September 2002. MAIN OUTCOME MEASURES: Patterns of medication use and safety, level of health care utilization, and health outcomes. RESULTS: A total of 104 patients reported taking a mean (+/- SD) of 6.7 +/- 3.8 medications. A total of 72 (69%) patients reported taking their medications correctly, and 90 (87%) reported finishing their medication course as prescribed. Medication refills were obtained by 75 (72%) patients, but of these patients, only 55 (73%) indicated that they obtained their refills on time. Adverse effects occurred in 25 (24%) patients, and 2 patients reported an allergic reaction. A total of 51 (49%) patients made unscheduled visits to their primary care physician, another health care facility, an emergency department, and/or were admitted to a hospital. Unscheduled visits occurred more often among nonadherent patients (59%) than adherent patients (44%), but not significantly so. In addition, 82 (79%) patients reported an improvement in health-related quality of life (QOL); 90 (87%) had a means of transportation to obtain medications; and 93 (89%) indicated that they would have to skip medications or give up necessities, if they were not enrolled in a subsidized prescription program. CONCLUSION: An improvement in self reported QOL and a high rate of medication adherence demonstrate support for the benefits of this and similar subsidized prescription drug programs. A high rate of additional health care utilization, especially among nonadherent patients, indicates an area for further analysis, program revisions, and/or patient education. PMID- 16381419 TI - Evaluation of accuracy of health studies reported in mass media. AB - OBJECTIVE: To evaluate communication of clinical research in the written media for completeness and accuracy. DESIGN: Observational assessment. SETTING: United States. PARTICIPANTS: Not applicable. INTERVENTIONS: Content of media articles discussing randomized controlled trials was assessed by three reviewers on the basis of the Consolidated Standards of Reporting Trials (CONSORT) criteria modified for the mass media. Reports from October 1 through December 31, 2002, published in the top two U.S. daily newspapers (USA Today and Wall Street Journal), weekly news magazines (Time and Newsweek), and daily news Web sources (CNN.com and MSNBC.com) and the corresponding published RCTs were analyzed. MAIN OUTCOME MEASURES: Total score and score in 10 specific content areas, leading to classification of coverage as poor, fair, or excellent. RESULTS: A total of 60 media reports discussing results of 25 RCTs appeared in these media during the study period. All reports were categorized as fair, and no content area was rated excellent. Several content areas received poor rankings in all and/or most media, including reporting of adverse effects, outcomes data, and statistical tests used. Media reports written by newswire services were rated more highly than were those prepared by nonnewswire services, but only 1 of 10 criteria had statistically significant differences. CONCLUSION: Mass media reports of RCTs are often incomplete. This type of reporting may misinform the lay public and may lead to questions about the applicability of the results to individual patients. PMID- 16381420 TI - Strategic planning by independent community pharmacies. AB - OBJECTIVES: (1) To assess the degree and level of use of the strategic planning process (none, partly, fully) by independent community pharmacy owners/managers and (2) to evaluate the relationships between independent community pharmacy owners/managers' level of strategic planning and indicators of pharmacy performance; including new and refill prescriptions filled, gross margin, rated patient care performance, rated dispensing performance, rated non-pharmacy performance, and rated financial performance. DESIGN: Cross-sectional study. SETTING: United States. PARTICIPANTS: Nationwide random sample of 1,250 owners/managers of independent community pharmacies. INTERVENTIONS: Mailed survey. MAIN OUTCOME MEASURES: Quality of strategic planning conducted; pharmacy performance measures. RESULTS: Only 141 of 527 (26.8%) usable responses indicated use of some (77 pharmacies, 54.6%) or all (64 pharmacies, 45.4%) of the seven steps typical of strategic planning. Significant associations were observed between the level of strategic planning use and all pharmacy performance variables assessed, including indicators such as greater numbers of new and refill prescriptions dispensed, gross margins, patient care performance, dispensing performance, non-pharmacy performance, and financial performance. CONCLUSION: Greater ratings of pharmacy performance were significantly associated with the level of strategic planning use. Respondents who fully used strategic planning had significantly higher indicators than partial users; respondents who partly used the process had significantly higher ratings than respondents who did not conduct strategic planning. PMID- 16381422 TI - Judging a book by its cover: the predicament with Canadian pharmacy advertisements and American consumers. PMID- 16381421 TI - Availability of and attitudes toward resources on alternative medicine products in the community pharmacy setting. AB - OBJECTIVE: To examine the availability of resources on dietary supplements in the community pharmacy setting and to assess the attitudes of community pharmacists toward these resources. DESIGN: Cross-sectional study. SETTING: Community pharmacies in New York and New Jersey that serve as experiential sites for senior student pharmacists at the Arnold & Marie Schwartz College of Pharmacy and Health Sciences, Long Island University. PARTICIPANTS: Preceptors or full-time pharmacists. INTERVENTION: Mailed survey. MAIN OUTCOME MEASURES: Frequency of use, availability of, and pharmacists' satisfaction with resources on alternative medicines, defined in the survey as any product including herbal remedies, vitamins, minerals, and natural products that may be purchased at a health food store, pharmacy, supermarket, alternative medicine store/magazine for the purpose of self-treatment. RESULTS: A total of 64 pharmacists characterized their frequency of use of resources on alternative medicines while formulating responses to questions as: never (n = 5; 7.8%), seldom (n = 31; 48.4%), often (n = 24; 37.5%), or always (n = 4; 6.3%). A total of 30 different resources were available to 40 respondents. The most commonly available resources were the PDR for Herbal Medicines (42.5%), The Review of Natural Products (20.0%), and the Web site of the National Center for Complementary and Alternative Medicine (12.5%). Of 54 respondents, 4 (7.1%) reported being completely dissatisfied with the available resources; 17 (31.5%) were somewhat dissatisfied; 25 (46.3%) were somewhat satisfied; and 8 (14.8%) were completely satisfied with the resources available to them. Topics the pharmacists most commonly wanted to see improved included safety (72.7%), interactions (70.9%), and uses (69.1%). CONCLUSION: Community pharmacists do not use information resources on alternative medicine products frequently, and many are not satisfied with the resources available to them. PMID- 16381423 TI - Pharmacists' activities supporting medication adherence: a regional survey. PMID- 16381424 TI - Edward Parrish (1822-1872): pioneer ethicist. PMID- 16381425 TI - Pertussis update: focus on adolescents. PMID- 16381426 TI - New drugs: pregabalin and omega-3-acid ethyl esters. PMID- 16381427 TI - OTC product: Monistat Soothing Care for vaginial discomfort. PMID- 16381429 TI - Dental emergencies at school--how to treat and prevent oral emergencies. PMID- 16381430 TI - Managing hypoglycemia in the school setting. PMID- 16381431 TI - What's the latest buzz on caffeine? PMID- 16381432 TI - Standing orders for allergy symptoms and anaphylaxis in the K through 12 school setting. PMID- 16381433 TI - The musculoskeletal examination: how and when to evaluate. PMID- 16381434 TI - "It's a celebration"! PMID- 16381435 TI - The wonderful world of children. PMID- 16381436 TI - Dental caries disease in children: management by risk assessment. PMID- 16381437 TI - Medical advances in pediatric health care: what dentists need to know. PMID- 16381438 TI - Management of pediatric permanent tooth avulsion. PMID- 16381439 TI - Glass ionomer cement restorations for contemporary pediatric dentistry. PMID- 16381440 TI - Status of amalgams in pediatric dentistry: pros and cons. PMID- 16381441 TI - Treating very young patients with conscious sedation and medical immobilization: a Jewish perspective. PMID- 16381442 TI - The stainless steel crown revisited. PMID- 16381443 TI - HIV and AIDS in children and adolescents. PMID- 16381444 TI - Making some sense of child behavior. PMID- 16381445 TI - Practice management for the fee-for-service pediatric dentist. AB - In conclusion, good practice management in pediatric dentistry is not sporadic or a series of random, uncoordinated activities. Instead, it encompasses the philosophy of the practice, is centered around the patient and is a fluid, consistent management function. It begins with the pediatric dentist as the leader and continues in all phases of daily activity. It is a well-developed and highly functioning system. It results in staff and patient loyalty and high patient satisfaction. The challenge is to make the practice an ongoing, continuously active, evolutionary process for many years to come. PMID- 16381446 TI - We have made a difference! PMID- 16381447 TI - [Insurance medicine--a challenge in the national and international context]. PMID- 16381448 TI - [Mitral valve stenosis (in adults)]. PMID- 16381449 TI - [Between selection and integration: the "double history" of insurance medicine in the 19th and 20th century]. PMID- 16381450 TI - [Disability management and occupational health systems]. PMID- 16381451 TI - [The semiotics of fibromyalgia and related somatoform disorders]. PMID- 16381452 TI - [Neurophysiological aspects of chronic pain]. PMID- 16381453 TI - [On the medicolegal assessment of somatoform disorders]. PMID- 16381454 TI - [Insurance and late-onset genetic disorders--fears and misunderstandings]. PMID- 16381455 TI - [Application of "3S" technologies in continuous usage of traditional Chinese medicine resources]. AB - This paper introduced development of 3S technologies, i.e., remote sensing (RS), geographic information system (GIS), and global positioning system(GPS), and drew the conclusion that 3S technologies can be used widely in resources dynasty survey, ecology study, and geoherbs study and classification about traditional Chinese medicine resources. PMID- 16381456 TI - [The new progress of the study about volatile oil of the angelica]. AB - To summarize the new progress of the study about volatile oil of the angelica, including the distillable methods, the analysis of the chemical components, the pharmacological effects and the clinical applications. We tracked and searched the correlative references and study reports about volatile oil of the angelica in CNKI data base(1994-2004) and Medline data base (1997-2004). We summarized and compared the different distillable methods of volatile oil of the angelica, meanwhile we summarized many study reports about the analysis of the chemical components of volatile oil of the angelica and it's pharmacological effects, including the toxicity of the volatile oil and it's effects on the uterus smooth muscle, cardiovascular system, respiratory system, central nerve system and immune system. Finally we summarized the clinical application of the volatile oil of the angelica. There are three distillable methods of volatile oil of the angelica . The harvest efficiency of volatile oil is different with different distillable methods. The chemical components are very complicated and the new chemical components are separated and identified. The volatile oil has bidirectional effects on the uterus smooth muscle. It can inhibit the contraction of the uterus smooth muscle induced by different mechanisms. Meanwhile it can depress the blood pressure and ameliorate the cardiac ischemia. The volatile oil can resist the arrhythmia and asthma, restrain the central system, improve the immune function. Nowadays the volatile oil of the angelica is applied to therapy the dysmenorrhea and disorder of the catamenia. The chemical components of the volatile oil of the angelica are very complicated, moreover the pharmacological effects of the volatile oil are comprehensive. People make the new progress of the study about volatile oil of the angelica. PMID- 16381457 TI - [The quality specification of grape seed extract]. AB - This paper reiviewed the current situation of quality control of grape seed extract in domestic and international market. Considering the fact that there is no national or industrial technical specifications established for the extract product, the authors suggested that two sets of quality specifications should be established for the grape seed extract. The two sets of specifications are: the high purity grape seed extract should contain polyphenol NLT 95%, monomer NLT 10%; and the grape seed extract with ordinary quality should have a procyanidolic value NLT 95, and monomer NLT 6%. PMID- 16381458 TI - [Genetic relationships among Fructus Aurantii cultivars revealed by ISSR]. AB - OBJECTIVE: A total of 27 samples belonging to 5 cultivars of Fructus Aurantii (Citrus aurantium), i. e. cv. Xiucheng, cv. Xiangcheng, cv. Lecheng, cv. Jizicheng, and cv. Youzicheng, collected at Changfu and Huanggang, Zhangshu City, Jiangxi Province, were assayed to reveal the genetic relationship among the cultivars and the accordance between morphological and molecular markers. METHOD: Cultivar identification was based on morphology and cultivar relationship was based on Inter-simple sequence repeats (ISSR). RESULT: Twenty out of 40 ISSR primers screened generated 392 loci across all 27 samples with 315 informative loci. The UPGMA dendrogram showed that samples within cv. Xiucheng and cv. Xiangcheng from Changfu were closely related. However, samples of cv. Lecheng, cv. Jizicheng and cv. Youzicheng from Huanggang, or cv. Xiucheng and cv. Xiangcheng from both Changfu and Huanggang did not exhibited close relationships within each cultivars. CONCLUSION: Based on morphology the same cultivar grown in different plantations, or even within a single plantation sometimes do not show close genetic relationship, indicating diverse origin of the cultivars. Synonyms or homonyms are believed to common phenomenon in Fructus Aurantii production. To solve the problem ISSR markers can serve a kind of molecular markers which are preferable to partition genetic variations within and between cultivars and to establish genetic relationships among them. PMID- 16381460 TI - [Study on HPLC fingerprint and quality control of Propolis]. AB - OBJECTIVE: To study on HPLC fingerprint of Propolis and to control it's quality. METHOD: The chromatographic fingerprints of seven samples from different producing areas were determined by RP-HPLC. RESULT: The chromatograms of Propolis from different producing areas were very similar. CONCLUSION: The quality of Propolis can be controlled by determination the HPLC fingerprint. PMID- 16381459 TI - [Effects of cultivation technique on yield and favonoid content of Chrysanthemum flower (Qiju) grown in Hebei]. AB - OBJECTIVE: To study the effect of cultivation techniques on the flower yield flavonoid content in Chrysanthemum flower grown in Hebei. METHOD: Studied on flowers yield and three factors (transplanting date and plant density and fertilizer quantity) were examined in field experiment at 4 treatments levels. RESULT AND CONCLUSION: The best results were obtained at following conditions: diammonium phosphate 300 kg x hm(-2) and potassium sulfate 150 kg x hm(-2) fertilized before transplanting, transplanting at the first ten days of May and the spacing 40 cm x 40 cm. PMID- 16381461 TI - [Study on the pretreatment of the series of HPD macroporous adsorption resin]. AB - OBJECTIVE: To develop a method for the pretreatment of the series of HPD macroporous adsorption resin (MAR). METHOD: The effect of 2% NaOH to pretreatment of the series of HPD MAR was studied by gas chromatography and UV spectrometery. RESULT: The eluting effect by ethanol was better after the MAR was marinated and eluted by 2% NaOH. CONCLUSION: The pretreatment of the series of HPD MAR was: after marinated and eluted by 2% NaOH, the series of HPD MAR were eluted by ethanol at 2 BV x h(-1) at 60 degrees C for 3-4 bed volumes (BV). At then the UV absorbance of eluate was 0.2-0.5. Benzene was not detected and the other residues less than 10 mg x L(-1) in eluted-MAR by GC. And the MARs match to the standard of medicine. PMID- 16381462 TI - [Study on chemical constituents in total saponin from Trigonella foenum-graecum]. AB - OBJECTIVE: To study the chemical constituents in the total saponin from Trigonellf foenum-graecum. METHOD: The compounds were isolated by column chromatography on macroporous resin and silica gel and elucidated by physical and chemical evidences and spectroscopic analysis. RESULT: Two compounds were obtained and identifiedas methyl-protodioscin and methyl-protodeltonin. CONCLUSION: Methyl-protodioscin and methyl-protodeltonin were isolated from this plant for the first time. PMID- 16381463 TI - [Studies on flavonoids from herb of Artemisia ordosica]. AB - OBJECTIVE: To study the chemical constituents of Artemisia ordosica. METHOD: The chemical constituents were isolated and repeatedly purified on silica gel column and the structures were elucidated by the NMR spectra and physico-chemical properties. RESULT: Eight flavonoids were obtained and identified as 5-hydroxy-7, 4'-dimethoxyflavanone, 5-hydroxy-7, 4'-dimethoxyflavone, 5, 4'-dihydroxy-7 methoxyflavone, 5, 7-dihydroxy-6, 4'-dimethoxyflavone, 5, 4'-dihydroxy-7 methoxyflavanone, 5, 3', 4'-trihydroxy-7-methoxyflavanone, 5, 7-dihydroxy-3', 4' dimethoxyflavone and 5, 3', 4'-trihydroxy-7-methoxyflavone. CONCLUSION: All the compounds were obtained from A. ordosica for the first time. PMID- 16381464 TI - [Determination of catechin and its analogues in Spatholobus suberectus by RP HPLC]. AB - OBJECTIVE: To develop a method for determination of catechin and its analogues in Spatholobus suberectus. METHOD: Gallocatechin, catechin and epicatechin are used as reference substances. Chromatographic assay is performed on a C18 colum. The mobile phase is composed of methanol and acetic acid solution with gradient elution. Velocity of flow is 0.8 mL x min(-1) and the detection wavelength is 270 nm. RESULT: The calibration curves of gallocatechin, catechin and epicatechin are linear within the range of 0.397 microg to 1.986 microg, 0.404 microg to 2.019 microg and 0.405 microg to 2.024 microg. The recoveries of them are 97.2%, 100.8% and 98.8%, and the RSD are 1.7%, 2.4%, 2.4% (n = 5). CONCLUSION: The method is simple, rapid and suitable for the determination of catechin and its analogues in S. suberectus. PMID- 16381465 TI - [Identification of triacylglycerols in coix oil by high performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry]. AB - OBJECTIVE: To identify triacylglycerols in coix oil. METHOD: High performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry was used for identification. The experiment was operated under the conditions: spray voltage at 3 000 V, capillary temperature at 250 degrees C, APCI vaporizer temperature at 400 degrees C, and corona current of 4 microA. Sheath gas pressure (high purity liquid nitrogen) was 35 kPa. Mass spectra were obtained over the m/e range of 300 to 900 amu, scan duration of 1s and Q1 peak width at 0.7. The stationary phase was Zorbax Extend C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase: dichloromethane-acetonitrile (35:65), flow rate: 1 mL x min(-1); column temperature: 25 degrees C. RESULT: 12 triacylglycerols were identified by HPLC-MS method. CONCLUSION: The result can be used to identify the components in a fingerprint chromatogram of coix oil and its related injection product. PMID- 16381466 TI - [Studies on the changes of 5-hydroxymethyl furfuraldehyde content in Radix Rehmanniae steamed for different time]. AB - OBJECTIVE: To find the changing regularity of 5-Hydroxymethyl furfuraldehyde (5 HMF) content in Radix Rehmanniae steamed for different time. METHOD: HPLC method was used, with C18 column (6 mm x 200 mm, 5 microm), a mixture of methanol-water (15: 85) as the mobile phase. The flow rate was 1.0 mL x min(-1). The UV detection was setat 284 nm and the column temperature was set at 24 degrees C. RESULT: The linear range was from 0.01 to 0.08 microg, r = 0.999 9 (n = 5). The average recovery was 100.2% and RSD was 2.7% (n = 5). CONCLUSION: The 5-HMF content increases as the processing time prolongs in a certain time rang. PMID- 16381467 TI - [Effects of rhubarb on the immune substances secreted from intestine in mice]. AB - OBJECTIVE: To investigate the therapeutic mechanism of rhubarb in protecting the intestinal muco-membranous barrier in the mice. METHOD: Bal b/c mice were divided into 2 groups, gavaged with normal saline and 10% rhubarb decoction, respectively. The animals were killed after 24 hours after the treatments. The intestinal juice was collected after intestinal lavage and centrifuged for determination of IgA, total protein, C3, high density lipoprotein, type II PLA2 activity, and content of lysozyme. At the same time, 40 mg of small intestine were incised in each mouse. Reverse transcription polymerase chain reaction (RT PCR) and gel image analysis were performed to detect the content of the cryptdin gene expression. RESULT: The content of IgA, total protein, the C3, lysozyme, and the type II PLA2 activity in intestinal lavaged juice exhibited the statistical differences between the two groups (P < 0.05). There were no significant difference in the ontents of HDL, cryptdin-1 and cryptdin-4 gene expression between the two groups (P > 0.05). CONCLUSION: Rhubarb could increase secretion of several immune associated substances of the mucous membrane in normal intestine, indicating a possibility to abate the injury of intestine mucus resulted from severe stress induced by trauma, burn and shock. Through above mechanisms Rhubarb may also reduce the incidence of bacterial translocation and systemic inflammatory reaction syndrome (SIRS). PMID- 16381468 TI - [Study on anti-cancer components of Fig residues with supper critical fluid CO2 extracting technique]. AB - OBJECTIVE: To optimize Supper Critical CO2 extracting technical (SFE-CO2) methods for extraction of anti-cancer active components of Fig Residues and to investigate the anti-cancer effect of the extract in vitro and in vivo. METHOD: The anti-cancer activity of extracted compound was measured on U937,95D and AGS cancer cells in vitro by MTT method. The anti-cancer effect of the extraction of Fig Residues was studied on mice transplant liver cancer in vivo. RESULT: The SFE CO2 condition for extraction of the anti-cancer components of Fig Residues was optimized as follows: granularity was 100, the extraction pressure was 30 MPa, the extraction temperature was 45 degrees C, the extraction time was 6 h and the CO2 flux was 12 L x h(-1); The IC50 of anti-cancer active components of Fig Residues on U937, 95D and AGS cells were 70.125 microg x mL(-1), 127.957 microg x mL(-1), 116.000 microg x mL(-1); The anti-cancer active components of Fig Residues inhibited 49.3% of the transplanted liver cancer in the mice. CONCLUSION: The method for extracting the anticancer active components of Fig Residues is stable and reasonable, and the extract from Fig Residues is of the anticancer effect. PMID- 16381469 TI - [Protective effect of the pretreatment with Chuanxiong-phthalide A on the vascular endothelial cells impaired by the ischemia and reperfusion in isolated rats hearts]. AB - OBJECTIVE: The aim of this study was to investigate the protective effect of Chuanxiong-pathalide A on the injury of endothelial cell induced by ischemia and reperfusion in isolated rat hearts. METHOD: The isolated rat hearts were perfused under constant pressure with Chuanxiong-pathalide A at the concentrations of 0.012 5 mg x mL(-1), 0.025 mg x mL(-1) and 0.05 mg x mL(-1) within 10 min followed by a 10-min washout period before the induction of a 30-min normothermic global ischemia and 60 min reperfusion. RESULT: Pretreatment with Chuanxiong pathalide A produced a reduction in the incidence of reperfusion-induced ventricular fibrillation (VF) and ventricular tachycardia (VT). Pretreatment of the hearts with high dose of Chuanxiong-pathalide A (0.05 mg x mL(-1)) prior to the ischemia and reperfusion, reduced the incidence of reperfusion-induced VF and VT to 37.5% as compared with non-pretreated control group (P < 0.05). The duration of occurrence of VF and VT in the group pretreated with Chuanxiong pathalide A at dosages of 0.012 5 mg x mL(-1), 0.025 mg x mL(-1) and 0.05 mg x mL(-1) were (7.50 +/- 1.61), (1.64 +/- 0.67) and (1.06 +/- 0.70) min respectively, which were significantly shorter than (23.51 +/- 3.99) min in non pretreated control group; 0.05 mg x mL(-1) of Chuanxiong-pathalide A increased coronary flow in comparison with the control group. The content of SOD in the group pretreated with 0.05 mg x mL(-1) of Chuanxiong-pathalide A was (59.6 +/- 18.7) U x mg(-1), significant lower than (92.3 +/- 19.0) U x mg(-1) in the non pretreated group. The contents of LDH and MDA were (2 378.0 +/- 196.3) U x g(-1) and (12.1 +/- 1.3) nmol x mg(-1) in the non-pretreated group, which were much higher than the values of (1 669.4 +/- 192.5) U x g(-1) and (6.9 +/- 0.8) nmol x mg(-1) in the group pretreated with Chuanxiong-pathalide A respectively. In addition, enzymeimmunoassays showed an decreased production of IL-1beta and the ratio of TXB2/6-Keto-PGF1alpha. CONCLUSION: Our results show that chuanxiong pathalide A pretreatment can protect the endothelial function from the injury caused by ischemia and reperfusion. PMID- 16381470 TI - [Study on the anticancer activities of the Clematis manshrica saponins in vivo]. AB - OBJECTIVE: To study the anticancer activity of the Clematis manshrica saponins in vivo. METHOD: Anticancer activities were tested in mice with experimental tumor (S180, HepA and P388) in vivo. RESULT: The Clematis manshrica saponins showed a significant anticancer activities on Sarcoma-180, HepA and P388 implanted in mice. In S180 sarcoma, the average tumor inhibition rates were 42.78%, 52.06% and 58.25% (P < 0.05-0.01) respectively; The mean inhibition rates were 37.44%, 52.05% and 59.36% (P < 0.05-0.001) in Hep A tumor separately; while in P388 tumor, the mean inhibition rates were 34.50%, 46.78% and 54.39% (P < 0.05-0.01), respectively. CONCLUSION: The results indicate that Clematis manshrica has obvious antitumor effects against various transplanted tumor in mice. PMID- 16381471 TI - [Effect on pharmacokinetics of pseudo-ephedrine in human plasma among herbs in Mahuang decoction]. AB - OBJECTIVE: To develop a GC-MS method for the determination of pseudo-ephedrine in human plasma and study the mutative rules of the pharmacokinetic parameters of the pseudo-ephedrine in different compositions. METHOD: Orthogonal design was used to determine 8 compositions from Mahuang decoction (MHD). The healthy volunteers were divided into groups in random, each group including 8 men. After taking the medicine orally, vena blood would be taken out at different time. Determining pseudo-ephedrine in plasma and plotting the concentration-time curve, the pharmacokinetic parameters of each composition were calculated by WinNonlin 4.0.1. The statistical analysis of the pharmacokinetic parameters was proceeded by SPSS 10.0. RESULT: All concentration-time curves were adequately modeled by one compartment, first order absorption model, no lag time. Some parameters of pseudo-ephedrine showed significant variance (P < 0.05) in different compositions. The statistic results showed interactions between drugs in MHD (P < 0.05). CONCLUSION: The other herbs erds of MHD have some certain extent effects to the pharmacokinetic parameters of pseudo-ephedrine in the dominant drug. PMID- 16381472 TI - [The pharmacokinetics of rhein in 12 healthy volunteers after oral administration of rhubarb extract]. AB - OBJECTIVE: To study the pharmacokinetics of rhein in 12 healthy volunteers after oral administration of rhubarb extract. METHOD: The blood sample were obtained at 0,0.083,0.5,1,1.5,2,3,4,5,7,10 h after a single dose oral administration of rhubarb (50 mg x kg(-1)). The plasma rhein concentration was determined by HPLC. The pharmacokinetics of rhein were analysed by 3P97 program. RESULT: The absorption of rhein was very fastafter oral administration of rhubarb extract in the healthy volunteers. The main pharmacokinetic parameters of rhein were C(max) (3.20 +/- 1.08) microg x mL(-1); t(max) (1.03 +/- 0.41) h; t(1/2alpha) (0.21 +/- 0.02) h; t(1/2beta) (2.68 +/- 1.09) h; MRT(5.31 +/- 1.78) h; AUC(0-infinity) (1 573.08 +/- 366.48) microg x mL(-1) min(-1), respectively. CONCLUSION: Rhein could be absorbed rapidly and its pharmacokinetics was consistent with two-compartment model. PMID- 16381473 TI - [Discussion on criterions of endanger and protection levels of traditional Chinese medicine herbs]. AB - To efficiently protect the endangered traditional Chinese medicine herbs is essential for the sustainable development of traditional Chinese medicine. On the bases of present species endanger and protection levels, problems in the traditional Chinese medicine herbs are analyzed. The endangered levels of traditional Chinese medicine herbs should refer to the standard of IUCN, and the protection levels should adopt qualitative and quantitative analysis based on the characteristic of traditional Chinese medicine herbs. Some qualitative and quantitative factors are discussed, some useful information was also provided for the establishment of protection levels of traditional Chinese medicine herbs. PMID- 16381474 TI - [IV Russian National Congress "Occupation and Health" (Moscow, October 25-27, 2005)]. PMID- 16381475 TI - [Health of able-bodied population in Russia]. AB - Dynamics of demographic parameters in Russia depicts marked negative trends. Mortality among able-bodied population after 1991 increased 2 times mostly due to accidents, poisonings and traumatism including suicides, and cardiovascular diseases. The authors consider parameters of occupational morbidity, occupational traumatism, transitory disablement morbidity and disability, demonstrate negative influence of occupational factors on life expectancy and children's health, suggest complex of priorities aimed to preservation of health in Russian population. PMID- 16381476 TI - [Evaluation of homeostatic disorders induced by radiation exposure]. AB - Possibilities of correction and prophylaxis. The authors presented results of study covering workers of radiation-dangerous plants of Siberian Chemical Enterprise, who underwent long-term exposure to outer and inner radiation in "low" doses range during the occupational activity. Target-systems for radiation effects were proved to be lipid peroxidation and antioxidant defence systems, cell energy metabolism--severity of disorders in which is correlated with degree of radiation exposure. PMID- 16381477 TI - [Analysis of health state in individuals exposed to thorium and chemical hazards in occupational environment]. AB - Clinical and epidemiologic study of chronic somatic discases in workers engaged into thorium production (main group) in short-term period revealed significantly higher prevalence of vegetative neurosis, asthenic state, chronic gastritis, arterial hypotension, chronic nasopharyngitis and higher risk of those conditions, if compared to reference group over the period since 1947 to 1959. These findings are caused by combined exposure to occupational hazards: outer and inner radiation due to long-lived thorium, thoron and their radioactive products, associated exposure to toxic chemicals. Chronic radiation sickness was diagnosed only in individuals who worked over the period since 1947 to 1959 and demonstrated hemopoietic disorders associated with functional neural system impairment. Better sanitary and hygienic work conditions and more efficient individual protective means ("Lepestok" respirator) resulted in reliably decreased risk of chronic somatic diseases among thorium production workers employed since 1960 to 1972. PMID- 16381478 TI - [Medical and biologic consequences of human chronic exposure to radiation]. AB - The author considered medical consequences of radiation accidents on Industrial Association "Mayak" (radioactive waste discharge into Techa river and accident in 1957). Results of longstanding observations helped to evaluate health state of people exposed to radiation and their descendants. PMID- 16381479 TI - [Some health parameters in personnel contacting highly enriched uranium]. AB - The authors analysed 20-years and longer observations over health state of personnel having occupational contact with highly enriched uranium. Findings are no determined effects as clinical manifestations of chronic uranium intoxication, presented by most frequently involved systems (hemopoietic, bronchopulmonary) and main organs accumulating uranium (liver, kidneys, bones). Long-term observations revealed malignancies in 11% of the examinees. Further medical observations over the personnel (risk group) are required to evaluate risk of aleatory effects in prolonged contact with various uranium compounds. PMID- 16381480 TI - [Theoretic evaluation of allowable time for evacuation from occupational zone of accidentally released uranium hexafluoride]. AB - Transdermal intake of uranium hexafluoride hydrolysis products is analysed. The authors present calculations of uranium and fluor amounts entering human body, in accordance with duration of stay in hazardous circumstances. PMID- 16381481 TI - [Parameters of physical and mental development of children and adolescents residing near enterprise for radioactive waste processing and stationary storage]. AB - Operation of enterprise for radioactive waste processing and disposal does not influence negatively physical and mental development of children and adolescents residing in the area under observation. In some parameters, physical and mental development of children and adolescents residing in the area under observation surpasses that in reference area--that could be due to social and economic peculiarities. PMID- 16381482 TI - [Immune state in uranium mine workers having respiratory diseases]. AB - The authors present analysis of immune disorders in miners with variable severity of occupational respiratory diseases. PMID- 16381483 TI - [Epidemiologic and radiobiologic features of neoplasms in respiratory and gastrointestinal tracts among patients who had occupational contact with uranium]. AB - Morbidity structure among workers engaged into main production of nuclear industry had prevailing lung cancer (1.93 per 1,000), gastric carcinoma (1.09) and colon cancer (0.72). The lung cancer patients demonstrated the highest concentrations of uranium in lung tissue (0.8313 g/g) and liver tissue (0.3548 g/g); those with gastric carcinoma and of reference group--in lung radix nodes (0.1855 and 0.3685 g/g respectively). PMID- 16381484 TI - Organochlorine pesticide levels in blood serum samples taken at autopsy from auto accident victims in Veracruz, Mexico. AB - Samples of human blood sera (N = 118) for the determination of organochlorine pesticide levels were obtained at autopsy from auto accident victims in Veracruz, Mexico, during the years 2000 and 2001. The presence of hexachlorobenzene (HCH), beta-hexachlorocyclohexane (beta-HCH), 2,2-bis(p-chlorophenyl)-1,1 dichloroethylene (p,p'-DDE), 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (p,p' DDT), and o,p'-DDT was confirmed by gas-liquid-electron-capture detection chromatography. During the years 2000 and 2001, the respective mean levels of (a) HCB, (b) beta-HCH, (c) p,p'-DDE, (d) o,p'-DDT, (e) p,p'-DDT, and (f) total DDT were (a) 2.1 ng/ml and 1.4 ng/ml, (b) 3.0 ng/ml and 3.6 ng/ml, (c) 21.1 ng/ml and 23.8 ng/ml, (d) 1.2 ng/ml and 0.8 ng/ml, (e) 3.3 ng/ml and 2.5 ng/ml, and, finally, (f) 25.4 ng/ml and 27.1 ng/ml, respectively. High levels of persistent organochlorine pesticides were--and continue to be--present in the blood of individuals who live in Mexico. Levels of insecticide metabolites (e.g., beta HCH, p,p'-DDE) in blood have increased during recent years (1997-2001), but levels of p,p'-DDT decreased in 2001 because the use of DDT for the control of malaria in Mexico was restricted. PMID- 16381485 TI - Gastrointestinal absorption of metallic mercury. AB - The absorption of mercury from the gastrointestinal systems of 7 subjects, of whom none had any amalgam fillings, was examined in this study. The authors obtained quantitative information about mercury concentration in plasma and duodenal fluid after the gastrointestinal systems of the subjects were exposed to liquid elemental mercury enclosed in rubber balloons (i.e., approximately 20 g of mercury), using a standard procedure followed for the sampling of bile. Plasma samples were collected prior to exposure, as well as up to 10 d following exposure, and duodenal fluid was collected 1 h, 2 h, 4 h, and 6 h during the intubation process. The authors studied the kinetics of dissolution in vitro by leaching elemental liquid mercury and mercuric chloride. The results of this study supported the hypothesis that metallic mercury is oxidized in the gastrointestinal tract. In addition, the authors determined that duodenal intubation, while using liquid metallic mercury in rubber bags, resulted in the diffusion of minor amounts of atomic elemental mercury through the rubber walls. The absorbed amount of mercury that reached the central circulation was comparable to a daily dose of mercury from dental amalgam in the amalgam-bearing population. PMID- 16381486 TI - Blood lead levels in preprimary school-age children in Nicosia, Cyprus, and their relationship with leaded soil dust exposure. AB - The authors conducted a cross-sectional study to determine blood lead levels in children who attended kindergarten schools and nurseries in Nicosia, Cyprus, and to correlate their findings with (a) home and school environments, (b) behavior of the children, and (c) socioeconomic characteristics. Capillary blood for lead assay was collected from March 2001 to September 2001 from children who lived and attended school in Nicosia. Children who lived and attended school in a rural setting served as controls. Parental questionnaires and interviews yielded information about socioeconomic background, environment, and children's habits and health. Overall findings indicated that children in Nicosia had mean blood lead levels similar to controls (i.e., <10 microg/dl)--the level of concern defined by the U.S. Centers for Disease Control and Prevention. The findings of our study likely represent high standards of hygiene adopted by parents and teachers, rather than knowledge embraced by parents and teachers about risks associated with lead exposure and sources of lead exposure. PMID- 16381487 TI - Time and temperature effects on the viability and infectivity of Cryptosporidium parvum oocysts in chlorinated tap water. AB - The authors compared the viability and infectivity of Cryptosporidium parvum oocysts in chlorinated tap water at various storage durations (i.e., 2 wk, 4 wk, 6 wk, or 8 wk) and at 2 cool temperatures (i.e., 10 degrees C and 4 degrees C), using in vitro (excystation) and in vivo (suckling mouse) methods. After 8 wk, mean oocyst excystation decreased to 33.4% and 26.7% at 10 degrees C and 4 degrees C, respectively. Suckling mice infectivity was higher after storage at 10 degrees C than after storage at 4 degrees C. These data suggest that Cryptosporidium parvum oocysts can survive and remain infectious for 8 wk in cool chlorinated tap water. PMID- 16381488 TI - A dried blood spot method to evaluate cholinesterase activity in young children. AB - Field methods are needed to detect and monitor the organophosphate pesticide exposure of young children. Twenty children, aged 11 to 18 mo, living in an agricultural community along the United States/Mexico border were enrolled in a pilot study investigating methods to detect pesticide exposure. Healthy children were recruited at pediatric clinics with the informed consent of their parents. Venous blood samples were collected from children twice, 4 wk apart. Cholinesterase activity was compared in whole heparinized venous blood with venous blood samples dried on filter paper. Although the amount of activity in the dried blood was consistently less than in the heparinized blood, the activity was significantly correlated: Spearman r = .6 (p = 0.01). This dried blood method may be used during field studies to evaluate changes in cholinesterase values in children over time. PMID- 16381489 TI - Association of outdoor air pollution with chronic respiratory morbidity in an industrial town in northern India. AB - A cross-sectional study was performed in one industrial (study) and one non industrial (reference) town in Punjab State, northern India. Ambient air quality samples were collected and analyzed each week for 2 yr. Subjects were 3,603 individuals >15 yr old who were interviewed and whose lung functions were measured spirometrically. Their biomarkers were categorized in terms of obstructive or restrictive defects. Levels of total suspended particulates, nitrogen oxides, sulfur oxides, carbon monoxide, and ozone were significantly higher in the study town than in the reference town. The prevalence of chronic respiratory symptoms (cough, phlegm, breathlessness, or wheezing) was 27.9 and 20.3% in the study and reference towns, respectively (p < 0.05). That of obstructive ventilatory defect was 24.9 and 11.8% (p < 0.05), respectively. Logistic regression analysis showed that residence in the study town was independently associated with chronic respiratory symptoms (odds ratio [OR] = 1.5; 95% confidence interval [CI] = 1.2, 1.8; p < 0.001) and spirometric ventilatory defect (OR = 2.4; 95% CI = 2.0, 2.9; p < 0.001) after controlling for other demographic effects. PMID- 16381490 TI - Increased risk of preterm delivery in women residing near thermal power plants in Taiwan. AB - In this article, the researchers studied the prevalence of preterm births for women living near thermal power plants. The prevalence of delivery of preterm birth infants was significantly higher among women living within 3 km of a thermal power plant than among women living within 3-4 km of a plant. After controlling for several possible confounders (including maternal age, season, marital status, maternal education, infant gender, and birth site), the adjusted odds ratio was 1.14 (95% confidence interval, 1.01-1.30) for delivery of preterm infants for women living close to the thermal power plants. These data provide further support for the hypothesis that air pollution can affect the outcome of pregnancy, although a semiecological study cannot confirm a direct causal relationship. PMID- 16381491 TI - Reduction in kidney cancer mortality following installation of a tap water supply system in an arsenic-endemic area of Taiwan. AB - Arsenic is the major risk factor for blackfoot disease, a peripheral vascular disease that has been endemic to the southwest coast of Taiwan for more than 50 yr because of the consumption of local artesian well water containing high levels of arsenic. Long-term arsenic exposure has been associated with kidney cancer mortality in a dose-response relationship. In the early 1960s, a tap water supply system was implemented in the blackfoot-endemic areas. After the mid-1970s, artesian well water was no longer used for drinking or cooking in the region. The authors examined whether kidney cancer mortality decreased after the elimination of arsenic exposure from artesian well water. Standardized mortality ratios for kidney cancer were calculated for the blackfoot-endemic area for the years 1971 2000. Study results showed that mortality from kidney cancer declined gradually during this time; therefore, the association of arsenic exposure with kidney cancer mortality was likely causal. PMID- 16381492 TI - Paying the price for going wireless. PMID- 16381493 TI - Keeping the airwaves safe and secure. Increasing access to Wi-FI networks requires stronger tools to protect them. PMID- 16381494 TI - Is there interference in the air? Wireless network users say radio frequency and electromagnetic interference still are concerns. PMID- 16381495 TI - Versatile PDAs pick up RFID, bar codes. PMID- 16381496 TI - Hospitals get a wireless makeover. PMID- 16381497 TI - Readers' perspectives. . . The HIPPA security rule has contributed to better protection of patient information. Do you agree or disagree? PMID- 16381498 TI - [Antibody development and frequency of events temporarily associated with measles and rubella vaccines among university health personnel in Jalisco]. AB - OBJECTIVE: To evaluate the seroprevalence, seroconversion, anamnesic response and events temporally associated with immune status pre and post immunization with measles and rubella vaccine in health personnel from a public University in Guadalajara, Mexico. MATERIAL AND METHODS: We carried out a prospective, longitudinal and comparative study from May to June 2000 among 120 healthy volunteers. Informed consent was obtained from all participants. We administered measles (Schwarz) and rubella (RA 27/3) vaccines. Weekly phone calls during six weeks were recorded from each volunteer to assess local and systemic events temporally associated with immunization non attributable to any other disease. Serum samples were obtained before and after vaccination in 75 volunteers. Antibodies against measles and rubella were measured by an enzyme immunoassay kit (Behring) with cut-off points of 300mUI/mL 8UI/mL respectively. Statistical analysis included mean, standard deviation and paired Student's t-test (P < 0.05). RESULTS: 105/120 participants (87.5%) were followed during 6 weeks. 87.4% were health personnel and 44.6% were males. The age range was 17-71 years (median = 21). We found a prior history of measles in 57.1%. Local events included pain, heat, redness and induration and were reported by 4/105. Systemic events such as fever, exanthema, pain in joints and arthritis was reported by 9/105. The proportion of study subjects with protective antibodies against measles and rubella prior to vaccination was 90.7 and 94.7% respectively. Both groups reached 100% after vaccination. CONCLUSIONS: The proportion of non-immune health personnel was low and similar to other reports. However, the potential spread of measles and rubella virus from a non-immunized infected health care provider could be amplified by the number of patients seen daily during the peak period of 3-5 days. The vaccination of health personnel should be encouraged. PMID- 16381499 TI - [Systemic hypertension after kidney transplantation: associated risk factors and influence on graft survival]. AB - Systemic hypertension after kidney transplant (HAPT) has been associated with a reduction in graft survival and increased morbidity and mortality of kidney transplant recipients. With the use of calcinuerin inhibitors, prevalence of HAPT has increased to 60-80%. The purpose of this study was to document the prevalence of HAPT in kidney transplant recipients attending the Instituto Nacional de Ciencias Medicas y Nutricion "Salvador Zubiran" associated risk factors and the effect of hypertension in long term graft survival. We retrospectively reviewed the clinical charts of all the patients that underwent kidney transplant from 1984 to 1994. The following risk factors were studied: age, gender, cause of renal failure, presence of hypertension before kidney transplant, histocompatibility, acute rejection episodes, chronic rejection, serum creatinine values and use of cyclosporine. We divided subjects in two groups: normotensive (NT) and hypertensive (HT). HAPT included 3140/90 mmHg blood pressure level observed at least during two consecutive evaluations or the use of antihypertensive medication. We analyzed 215 grafts from 205 patients (10 patients had two kidney transplants); mean age at transplant of 30 +/- 9 years, 131 subjects were female and 84 male. One hundred and eighty eight patients (88%) displayed pretransplant hypertension. The mean follow up was 56+/-32 months. In the postransplant period 152 (71%) were HT and 63 (29%) NT. The HT group had significantly higher blood pressure and serum creatinine values than the NT group (P < 0.001), in spite of an adequate blood pressure control in 65% of the patients from the HT group. The NT group displayed a higher graft survival than the HT group; 60 +/- 30 months vs. 51 +/- 32 months respectively (p<0.01). Multivariate analysis did not show any risk factors independently associated with the development of HAPT. The prevalence of HAPT in our series is similar to the one reported in the literature. During the postransplant period there was a reduction of hypertensive patients (88% pretransplant vs. 71% postransplant). HAPT is a significant risk factor associated with long term survival of the graft. PMID- 16381500 TI - [Primary central nervous system lymphoma in immunocompetent patients]. AB - Primary central nervous system lymphoma has been traditionally described in patients with immunodeficiency syndromes; however, there is an increasing number of immunocompetent patients with this type of tumor that have been reported recently. In this paper we have retrospectively analyzed 22 immunocompetent patients with a confirmed diagnosis of primary lymphoma of the brain. The mean age in this group was 65 years with a similar male/female ratio. The time of evolution of the clinical course was 80.4 days and it was mainly characterized by headache and focal neurological deficit. In four patients multiple lesions were observed, while the remaining presented single lesions mainly located in the periventricular area of the cerebral hemispheres. All patients were initially administered steroids and a stereotactic biopsy was performed. The majority of tumors were histologically classified as diffuse large cells and all of them showed a positive reaction to B-cells antigens on immunohistochemistry. All patients were treated with radiotherapy and in 10 of them, chemotherapy with methotrexate was also indicated. The mean survival rate was II months among patients treated with radiotherapy alone and increased to 36 months when chemotherapy was added. PMID- 16381501 TI - [Prognostic value of pre-B immunophenotype in early treatment response among acute pediatric lymphoblast leukemia patients]. AB - OBJECTIVE: To determine the prognostic value of preB immunophenotype and its variants on early treatment response among of acute pediatric lymphoblast leukemia. PATIENTS AND METHODS: A case-control study nested in a cohort was carried out with male and female patients 15 years and younger with recently diagnosed pre-B lymphoblast leukemia. A panel of B, T, monoclonal antibodies of the myelo-monocytic and megakaryocytic cell type was used. Response was assessed by bone marrow aspiration 14 days post treatment. RESULTS: 54 patients were included. The median age was 7 years (2 months - 14 years) median leukocyte count was 13,450/mm3 (1200-986,000/mm3). We identified 29 cases with late pre-B immune phenotype, 19 cases with common pre B and 6 cases with early preB immunophenotype. Eleven, patients also displayed myeloid antigens. A significant association (p=0.034) was found between early treatment response and the presence of myeloid antigens. No association was found between the pre-B immunophenotype, age and leukocyte count with early treatment response (p=0.264). CONCLUSIONS: We need to pay special emphasis on early treatment response in children with lymphoblast leukemia as our study did not corroborate the common finding that clinical factors and immune phenotype can be predictive factors. PMID- 16381502 TI - [Teaching and including HIV/AIDS in the curricula of a medical school sample in Mexico]. AB - BACKGROUND: The HIV/AIDS epidemic is apublic health problem that has had an impact on all health systems around the world. Mexico is no exception. Although it has been acknowledged that we have a concentrated epidemic, the problem continues to grow. OBJECTIVE: The objective of this study was to evaluate if the medical school curricula in Mexico included the topic of HIV/AIDS and to assess the level of knowledge medical students have about this disease. MATERIAL AND METHODS: Descriptive study of eight medical schools interviewing pre-graduate medical students. RESULTS: Only 15% of the curricula of private medical schools and 21.7% of public schools included the subject of HIV/AIDS. Although it is difficult to standardize this finding, an average of 8.8 hours was assigned to the study of HIV/AIDS. 90% of the professors involved in teaching the topic of HIV/AlDS hadno clinical experience in thefield CONCLUSION: HIV/AIDS is a low priority subject in our medical school curricula. Efforts must be placed to standardize and reinforce this highly important topic. PMID- 16381503 TI - [Intracellular calcium-dependent contractile effects of serotonin in the human umbilical artery]. AB - BACKGROUND: Absence of innervation is a hallmark of human umbilical vessels. Intervillous space blood flow is regulated by vasoactive substances and calcium dependent contractility, both in normal and pathological conditions such as preeclampsia-eclampsia. OBJECTIVE: To obtain additional information on the intracellular calcium contractile effects of serotonin in human umbilical arteries. MATERIALS AND METHODS: Umbilical arteries from normal pregnancies were dissected, cut in 5 mm rings and mounted in a temperature-controlled isolated organ chamber, using calcium-free Krebs solution. The contractile effects of serotonin, lantane, verapamil and cyclopiazonic acid were evaluated at different concentrations using a computer coupled biopac polygraph. RESULTS: No differences in response were observed in the presence and absence of intracellular calcium. The positive contractile effects observed with serotonin were significantly decreased with repeated stimulation. An increase in the basal tone of the vessel was observed after calcium supplementation was added to the solution. This effect was minimized in the presence of verapamil and lantane. The contractile effects of serotonin in the calcium-free solution were not affected by the presence of cyclopiazonic acid. CONCLUSIONS: Serotonin contractile effects in the human umbilical artery depend mainly on intracellular calcium levels which favor the gradual entrance of this ion over time. Calcium influx induced by serotonin is possible through L and Non-L channels apparently insensitive to ciclopiozonic acid. PMID- 16381504 TI - [Leptine participation in the development of liver steatosis and biliar lithiasis]. AB - Obesity increases significantly the risk of developing several common gastrointestinal diseases such as gallstone disease (GD) and hepatic steatosis (HS). Elsewhere we have shown a relationship between HDL cholesterol, cholesterol saturation index, and leptin in obese patients loosing weight. Furthermore, leptin plays an important role facilitating HS and possibly in the associated inflammatory process. The aim of this study was to investigate the relationship between GD and HS. The sample was comprised by patients attending the unit for check-up. Subjects with visible stones or HS by ultrasound (cases) were compared with healthy controls. Demographic and body mass index (BMI) were recorded. Plasma leptin, insulin and serum lipids and lipoproteins levels were measured by standard methods. A total of 317 subjects were included in this study. They were divided in four groups as follows: GD (n=100), HS (n=84), GD + HS (n=33) and controls (n=100). The control group was significantly older (GD, 52.6+/-11.6; HS, 49.8+/-11.1; GD +HS, 51.6+/-10.5; 57.1+/-7.4), p< 0.05. BMI was higher in the HS groups (28.7 +/- 2.8) and GD +EH (29.0 +/- 3.8) than in the GD (27.4 +/- 4.3) and control (27.0 +/- 3.1) group, p<0.05. The GD group displayed the highest leptin levels (13.7 241 8.1), P < 0.05, whereas insulin levels were similar in all groups. Since GD and HS subjects have high plasma leptin levels compared with controls, our results suggest that leptin plays an important role in the pathophysiology of GD and HS. PMID- 16381505 TI - [Early breastfeed weaning as a risk factor for acute appendicitis in children]. AB - OBJECTIVE: To determine if short term exclusive (EBF), combined breastfeeding (CBF), or an early start of other kind of food constitute risk factors for appendicitis in children. MATERIALS AND METHODS: By means of a case-control study, the mothers of 200 children up to 16 years with a diagnosis of acute appendicitis and 200 matched controls were interviewed regarding the duration of breastfeeding and the start of other foods. Data were analyzed to calculate chi square c2; odds ratio (OR) was used to calculate risk factors, confidence intervals (CI 95%) were also determined. RESULTS: There was no EBF in 52 cases (26%), or in 5 controls (2.5%) (p< 0.001). EBF lasted for 2.3 +/- 1.8 months in cases and 3 +/- 1.1 months in controls (p< 0.001). CBF lasted for 8.1 +/- 7.5 months in cases and 8.8 +/- 3.5 in controls (p< 0.001); ablactation was started at 4.4 +/- 1.3 months in cases and at 4.7+/-1.2 in controls (p< 0.05). The OR of developing appendicitis for non- EBF was 10.4 (CI 95% 4 - 26.5), for ablactation before 4 months 1.4, (CI 95% 0.9 - 2) and for CBF for less than 6 months 4.6, (CI 95% 2.3 - 8.3). CONCLUSIONS: The risk of developing appendicitis is higher among children who never received EBF and who were breastfed for less than 6 months. PMID- 16381506 TI - [Adipocitokines, adipose tissue and its relationship with immune system cells]. AB - Adipokines or adipocytokines are the proteins secreted by the adipose tissue. These bioactive molecules include proteins that modify insulin sensitivity (acylation-stimulating protein (ASP), TNF-alpha IL-6, resistin, leptin and adiponectin), and proteins that have known effects on vascularity (angiotensinogen and the plasminogen inhibitor protein PAI-I). Several studies have found a close relationship between adipocytes and immune cells as a consequence of evolutionary mechanisms that favor metabolic adaptation and survival under adverse conditions. It is known that adipokines contribute to the inflammation and insulin resistance present in obese individuals. The aim of this review is to analyze current information related to the physiology of the adipose tissue, with a special emphasis on the secretion of adipokines and their role in inflammation. We recommend that therapies addressing the treatment of obesity related disorders should focus on modifying the inflammatory process that originates in the adipose tissue. PMID- 16381507 TI - [Signal transduction, pillar of the neurobiological integration of memory. An alternative view to the cholinergic hypothesis]. AB - Neurophysiological, biochemical and molecular processes described in the integration of memory are closely related with neurotransmitters such as glutamate and serotonin (SHT) and with the function of calcium and potassium ion channels more than with cholinergic activity. Infact, glutamate and 5-HT receptors are closely related with Long-Term potentiation (L TP) processes, the mechanism by which memory is preserved throughout time. That is, the activation of the 5-HTI receptor triggers a transduction signal that after influencing nuclear cell activity, provokes several presynaptic changes, which leads to the displacement of magnesium from the postsynaptic area depolarizing the neuron and leading to the activation of N-methyl-D-aspartate receptors (NMDA). As a whole, this process contributes to the support and perpetuation of LTP, which consists of the following processes: LTPI that depends on protein kinase activity; LTP2 linked to translation of genes; and LTP3 closely related to genes transcription. On the opposite side but in perfect balance, we find the mechanism of Long-Term depression (LTD), which is triggered instead when the Ca+ +flow decreases in the presynaptic neuron activating the inhibitor-1 enzyme that promotes the dephosphorylation of a calmodulin-dependent protein kinasell and as a result, the inhibition of autophosphorylation and consequently of LTP too. Despite the widespread dissemination of the cholinergic hypothesis in Alzheimer's disease, memory build up rather than involving acetylcholine essentially depends on the participation of other neurotransmitters such as 5-HT and glutamate, which have not been adequately considered in the treatment of this disease. However, beyond neurotransmission, it is the cellular mechanism of autophosphorylation of several protein kinases, the process susceptible of being activated or controlled by the action of distinct substances. In such a case, it would be possible to exert some influence on gene expression improving perhaps, some of the physiopathological deficits that characterize memory disruption. PMID- 16381508 TI - [A sixteen year-old male with right calf pain and motor difficulty]. PMID- 16381509 TI - [ALK-1 positive anaplastic large cell lymphoma of the lung. Report of a pediatric case]. AB - Ki-1 anaplastic large cell non-Hodgkin lymphoma is a well recognized clinical entity. The main clinical feature includes lymphadenopathy with mediastinal sparing. In the extranodal disease, which occurs in approximately half of the cases, the skin is the most common site; bone marrow, lung and central nervous system are generally not involved. Anaplastic large cell lymphoma is more common among young people, in whom the prognosis is more favorable. Primary anaplastic large cell lymphoma of the lung is a rare clinical entity. Its clinical expression is similar to a high grade malignant lymphoma, and in most cases the diagnosis is made in advanced stages. We present a pediatric case with ALK-1 positive anaplastic large cell lymphoma of the lung. PMID- 16381510 TI - [Early diagnosis of a newborn with a mediastinal mass]. AB - Neuroblastoma is an embryonal tumour that evolves from the neural crest cell. This neoplasm may arise at any site in the sympathetic nervous system, including the brain, the cervical region, the posterior mediastinum, the para-aortic sympathetic ganglia, the pelvis, and the adrenal medulla. The clinical presentation in neonatal age is rare and the differential diagnosis includes congenital lung malformations, pneumoniae, atelectasia, etc. The case of a newborn with a mediastinal neuroblastoma is presented. This case illustrates how a patient with an X-ray image compatible with a thoracic tumor should be studied. PMID- 16381511 TI - [Mesial temporal sclerosis]. PMID- 16381512 TI - [The chimpanzee genome]. PMID- 16381513 TI - [Uncommon metastases in renal carcinoma]. AB - INTRODUCTION: Renal clear cell carcinoma (RCCCa) represents 2-3% of the neoplasms in the adult population, and can be metastastatic in up to 30% at the time of diagnosis. In these cases its mortality can be of up to 99% in 5 years, due to the lack of response to current treatments, unless it is a single, surgically resectable metastasis. Currently, there is no explanation for its routes of progression, it even seems to be non existent based on the rare tumor implant sites. OBJECTIVE: To report unusual metastasis from RCCCa and a review of the literature. MATERIAL AND METHODS: We reviewed retrospectively 15 years of patient records diagnosed with metastasic CaRCC different from: local lymph nodes, lung, bone or liver. We identified 11 cases: brain (2), duodenum (2), tonsils, thyroid, esophagus, diaphragm, testicle, adrenal gland and soft tissue of the knee. CONCLUSION: A description of this and other literature series with rare metastasis was done, as well as an analysis of possible routes and mechanisms responsible for the unusual tumor progression in RCCCa. We need to continue investigating the dissemination pathways and other cellular conditions involved in metastastatic kidney cancer in order to better understand and improve the treatment of patients in advanced stages. PMID- 16381514 TI - Far-field diffraction patterns of circular sectors and related apertures. AB - In studies of scalar diffraction theory and experimental practice, the basic geometric shape of a circle is widely used as an aperture. Its Fraunhofer diffraction pattern has a simple mathematical expression easily determined by use of the Fourier-Bessel transform. However, it may require considerable mathematical effort to determine the far-field diffraction patterns of aperture shapes related to the circular geometry. From a computational point of view, the mathematical difficulties posed by other aperture geometries as well as more general apertures with irregular shapes can be surpassed by means of optical setups or fast numerical algorithms. Unfortunately, no additional insight or information can be obtained from their exclusive application, as would be the case if mathematical formulas were available. The research presented here describes the far-field diffraction patterns of single-sector apertures as well as their extension to double symmetrical sectors and to sector wheels formed by interleaved transparent sectors of equal angular size; in each case, full or annular sectors are considered. The analytic solutions of their far-field amplitude distribution are given here in terms of a series of Bessel functions, some interesting properties are deduced from these solutions, and several examples are provided to illustrate graphically the results obtained from approximate numerical computations. Our results have been verified numerically with the fast-Fourier-transform algorithm and experimentally by means of a spherical wavefront-single-lens Fourier-transform architecture. PMID- 16381515 TI - Four-dimensional dynamic flow measurement by holographic particle image velocimetry. AB - The ultimate goal of holographic particle image velocimetry (HPIV) is to provide space- and time-resolved measurement of complex flows. Recent new understanding of holographic imaging of small particles, pertaining to intrinsic aberration and noise in particular, has enabled us to elucidate fundamental issues in HPIV and implement a new HPIV system. This system is based on our previously reported off axis HPIV setup, but the design is optimized by incorporating our new insights of holographic particle imaging characteristics. Furthermore, the new system benefits from advanced data processing algorithms and distributed parallel computing technology. Because of its robustness and efficiency, for the first time to our knowledge, the goal of both temporally and spatially resolved flow measurements becomes tangible. We demonstrate its temporal measurement capability by a series of phase-locked dynamic measurements of instantaneous three dimensional, three-component velocity fields in a highly three-dimensional vortical flow-the flow past a tab. PMID- 16381516 TI - Projection method of resolving ambiguities by determining the order of colors in moire fringes. AB - From the scientific literature there are some well-known methods used to resolve the arising problem of ambiguities in moire patterns. The main disadvantage of these methods is that for image processing one must use several patterns. It means that it is impossible to work with dynamic objects in a real-time operation mode. We present a projection method to resolve ambiguities by determining the order of colors in moire patterns. This method is based on the application of color rulings to gratings in a projection moire technique. PMID- 16381517 TI - Long-working-distance incoherent-light interference microscope. AB - We describe the design and operation of a long-working-distance, incoherent light interference microscope that has been developed to address the growing demand for new microsystem characterization tools. The design of the new microscope is similar to that of a Linnik interference microscope and thus preserves the full working distance of the long-working-distance objectives utilized. However, in contrast to a traditional Linnik microscope, the new microscope does not rely on the use of matched objectives in the sample and the reference arms of the interferometer. An adjustable optical configuration has been devised that allows the total optical path length, wavefront curvature, and dispersion of the reference arm to be matched to the sample arm of the interferometer. The reference arm configuration can be adjusted to provide matching for 5x, 10x, and 20x long-working-distance objectives in the sample arm. In addition to retaining the full working distance of the sample arm objectives, the new design allows interference images to be acquired in situations in which intervening windows are necessary, such as occur with packaged microsystems, microfluidic devices, and cryogenic, vacuum, or environmental chamber studies of microsystem performance. The interference microscope is compatible with phase-shifting interferometry, vertical scanning interferometry, and stroboscopic measurement of dynamic processes. PMID- 16381519 TI - Polymer-based flexible microlens arrays with hermaphroditic focusing properties. AB - Polymer microlens arrays with hermaphroditic focusing behaviors are demonstrated. Each microlens in an arrays exhibits either converging or diverging focus, depending on the polarization direction of the incident light. A polymer film with patterned microlens arrays is flexible, lightweight, and ultrathin (approximately 50 microm). Details of the lens structure, device fabrication, and lens performance are described. PMID- 16381518 TI - Signal-to-noise ratio study of full-field fourier-domain optical coherence tomography. AB - We report a new approach in optical coherence tomography (OCT) called full-field Fourier-domain OCT (3F-OCT). A three-dimensional image of a sample is obtained by digital reconstruction of a three-dimensional data cube, acquired with a Fourier holography recording system, illuminated with a swept source. We present a theoretical and experimental study of the signal-to-noise ratio of the 3F-OCT approach versus serial image acquisition (flying-spot OCT) approach. PMID- 16381520 TI - High-temperature stability multilayers for extreme-ultraviolet condenser optics. AB - We investigate the thermal stability of Mo/SiC multilayer coatings at elevated temperatures. Transmission electron microscopy and x-ray diffraction studies show that, upon annealing, a thermally induced structural relaxation occurs that transforms the polycrystalline Mo and amorphous SiC layers in as-deposited multilayers into an amorphous Mo-Si-C alloy and crystalline SiC, respectively. After this relaxation process is complete, the multilayer is stable at temperatures up to 400 degrees C. PMID- 16381521 TI - Optical characterization of ophthalmic lenses by means of modulation transfer function determination from a laser speckle pattern. AB - A method is presented for measuring the modulation transfer function of ophthalmic lenses by use of the generation of laser speckle with an integrating sphere. The measurements are performed with a rectangular double-slit aperture positioned at the output port of the integrating sphere. The distance between the lens and the detector determines the spatial frequency being tested; therefore high frequencies are tested close to the lens and low frequencies are tested far away from the lens. We can conclude that the double-slit method can be a versatile technique for comparing the optical quality of ophthalmic lenses from different makers. PMID- 16381522 TI - Effect of illumination in an integral imaging system with large depth of focus. AB - We present the characteristics of integral imaging systems with large depth of focus (DOF) by use of two kinds of illumination: plane illumination and diffusing illumination. For each system, we perform ray analysis based on ray optics. To check the visual quality through optical experiments, we use an average image of observed images picked up at various positions within a large DOF. The synthesized elemental images for a three-dimensional (3-D) object with two character patterns were displayed in an optical system and its reconstruction experiments are performed. Experimental results show that use of diffusing illumination can improve visual quality of reconstruction 3-D images in depth priority integral imaging. PMID- 16381523 TI - Stressed liquid-crystal optical phased array for fast tip-tilt wavefront correction. AB - A liquid-crystal optical phased-array technology that uses stressed liquid crystals provides a new type of tip-tilt wavefront corrector. It demonstrates a very fast time response (10 kHz) and high beam-steering efficiency (approximately 91%). The new technology presented here will allow for a nonmechanical, high speed correction with simple device construction. PMID- 16381524 TI - Random number generation based on the time of arrival of single photons. AB - We report the demonstration of a new type of true random number generator based on the random distribution of the time interval between photons from a single photon-like source. The experimental setup is simple and robust against mechanical and temperature disturbances. With improved detector resolution and efficiency, the random number bit rate could be increased by more than an order of magnitude to satisfy practical requirements. PMID- 16381525 TI - Radiative transfer code SHARM for atmospheric and terrestrial applications. AB - An overview of the publicly available radiative transfer Spherical Harmonics code (SHARM) is presented. SHARM is a rigorous code, as accurate as the Discrete Ordinate Radiative Transfer (DISORT) code, yet faster. It performs simultaneous calculations for different solar zenith angles, view zenith angles, and view azimuths and allows the user to make multiwavelength calculations in one run. The Delta-M method is implemented for calculations with highly anisotropic phase functions. Rayleigh scattering is automatically included as a function of wavelength, surface elevation, and the selected vertical profile of one of the standard atmospheric models. The current version of the SHARM code does not explicitly include atmospheric gaseous absorption, which should be provided by the user. The SHARM code has several built-in models of the bidirectional reflectance of land and wind-ruffled water surfaces that are most widely used in research and satellite data processing. A modification of the SHARM code with the built-in Mie algorithm designed for calculations with spherical aerosols is also described. PMID- 16381526 TI - Definition, meaning, and measurement of the polarization extinction ratio of fiber-based devices. AB - We clarify the definition of the polarization extinction ratio-also called polarization cross talk--of fiber-based devices. Its strong wavelength dependence, even for simple devices such as a single-fiber patchcord, is highlighted. Thus white sources may not be used for most measurements. We also explain the weakness of measurements with a rotating polarizer and a monochromatic source. Only a polarimeter may be used if accuracy is required. We report measurements of connections (including two connectors and one mating sleeve) and show the importance of the mating sleeve in the result. Finally, we define the validity domain of the standardized method, which uses both a white source and a rotating polarizer. PMID- 16381527 TI - Modeling and measurement of temperature sensitivity in birefringent photonic crystal holey fibers. AB - We analyzed theoretically the spectral dependence of polarimetric sensitivity to temperature (KT) and the susceptibility of phase modal birefringence to temperature (dB/dT) in several birefringent photonic crystal holey fibers of different construction. Contributions to dB/dT related to thermal expansion of the fiber dimensions and that related to temperature-induced changes in glass and air refractive indices were calculated separately. Our results showed that dB/dT depends strongly on the material used for manufacturing the fiber and on the fiber's geometry. We demonstrate that, by properly designing the birefringent holey fiber, it is possible to reduce its temperature sensitivity significantly and even to ensure a null response to temperature at a specific wavelength. Furthermore, we show that the temperature sensitivity in a fiber with arbitrary geometry can be significantly reduced by proper choice of the glass used in the fiber's manufacture. We also measured the polarimetric sensitivity to temperature and identified its sign in two silica-air fibers. The experimental values are in good agreement with the results of modeling. PMID- 16381528 TI - Study of transmission response of edge filters employed in wavelength measurements. AB - The ratiometric wavelength-measurement system is modeled and an optimal design of transmission response of the employed edge filter is demonstrated in the context of a limited signal-to-noise ratio of the signal source. The corresponding experimental investigation is presented. The impact of the limited signal-to noise ratio of the signal source on determining the optimal transmission response of edge filters for a wavelength-measurement application is shown theoretically and experimentally. PMID- 16381529 TI - Wavelength references for interferometry in air. AB - Cavity-mode wavelengths in air are determined by measuring a laser's frequency while it is locked to the mode in vacuum during a calibration step and subsequently correcting the mode wavelength for atmospheric pressure compression, temperature difference, and material aging. Using a Zerodur ring cavity, we demonstrate a repeatability of +/- 2 x 10(-8) (3sigma), with the wavelength accuracy limited to +/- 4 x 10(-8) by knowledge of the absolute helium gas temperature during the pressure calibration. Mirror cleaning perturbed the mode frequency by less than deltav/v approximately 3 x 10(-9), limited by temperature correction residuals. PMID- 16381530 TI - Fiber ring laser with a feedback mirror. AB - We describe the spectral and power features of a ytterbium-doped double-clad photonic crystal fiber laser that is operated in a ring configuration with an external mirror that feeds back only one of its two output beams. We compare the operation of the laser with and without an external feedback mirror. We find that the feedback mirror reduces significantly the spectral and power fluctuations. It is also responsible for an interesting spectral phenomenon: The laser frequency is drifting periodically over 9 nm at a rate of 2 nm/s from a short wavelength to a longer wavelength and vice versa. PMID- 16381531 TI - Single-cavity, multiline laser system with large coherence length. AB - We present a new approach of generating the several wavelengths required for color holography with coherence lengths in the range of several meters. Our proposed laser system consists of an argon-ion laser, which is equipped with broadband optics. Its main lasing lines include 457, 488, and 514 nm. Sufficient coherence length is achieved by means of an intracavity etalon. We report single frequency operation at several competing wavelengths and the successful recording of multicolor holograms with the described laser system. PMID- 16381532 TI - Development of a 756 nm, 3 W injection-locked cw Ti:sapphire laser. AB - We have developed a 756 nm, 3 W single-frequency cw Ti:sapphire laser by using the technique of injection locking. A cw Ti:sapphire laser in a ring-type configuration was forced to lase unidirectionally by use of an optical diode to prevent a high-power backward laser from disturbing the injection laser. A master laser was amplified by a broad-area laser diode and coupled into a single-mode fiber to generate a 50 mW injection laser with a Gaussian beam profile, which was enough to lock the Ti:sapphire laser at full power of 3 W. Such a high-power single-frequency Ti:sapphire laser enables a watt-level blue or near-ultraviolet single-frequency laser to be generated by frequency doubling. PMID- 16381533 TI - Role of the stimulated-emission rate in the photostability of solid-state dye lasers. AB - The lasing and photostability characteristics of Rhodamine 6G and Pyrromethene 567 dyes dispersed in polymeric host materials have been investigated as a function of the intensities of incident pump and signal beams in a longitudinally pumped dye laser in an oscillator-amplifier configuration. A substantial reduction in the rate of photodegradation was observed under lasing conditions and with increasing signal intensity in a dye amplifier, establishing that the service lives of these materials improve with an increase in the rate of stimulated emission. We observed approximately 62% amplifier efficiency at 2 Hz operation and 10% reduction in amplifier efficiency at 10 Hz operation after exposure of 72,000 pulses by use of a Pyrromethene disk. PMID- 16381534 TI - Correction of spherical and azimuthal aberrations in radially polarized beams from strongly pumped laser rods. AB - Spherical aberration of heavily pumped Nd:YAG rods was corrected by use of spherical relay optics selected to add conjugate amounts of aberration. Wavefront measurements showed elimination of spherical aberration. Correction of spherical aberration allowed 250 W of power to be generated in a radially polarized, birefringence-free oscillator (60% more power than without correction). Scale-up of wavefront maintenance was demonstrated in a two-rod amplifier module (6.3 kW electrical pump power). Radial polarization and spherical aberration correction together eliminated the main aberrations in uniformly pumped rod-based lasers. Rotating adjacent pump chambers substantially reduced the multifold aberrations induced by nonuniformity of the azimuthal pumps. DeltaM2 in the radially polarized beam was 0.3 and 1.4 with and without aberration correction, respectively, in each two-pump chamber module. Analysis predicts further improvements when higher-order aberration correction is applied. PMID- 16381535 TI - Radiometric calibration of SeaWiFS in the near infrared. AB - The radiometric calibration of the Sea-Viewing Wide-Field-of-View Sensor (SeaWiFS) in the near infrared (band 8, centered on 865 nm) is evaluated by use of ground-based radiometer measurements of solar extinction and sky radiance in the Sun's principal plane at two sites, one located 13 km off Venice, Italy, and the other on the west coast of Lanai Island, Hawaii. The aerosol optical thickness determined from solar extinction is used in an iterative scheme to retrieve the pseudo aerosol phase function, i.e., the product of single scattering albedo and phase function, in which sky radiance is corrected for multiple scattering effects. No assumption about the aerosol model is required. The aerosol parameters are the inputs into a radiation-transfer code used to compute the SeaWiFS radiance. The calibration method has a theoretical inaccuracy of plus or minus 2.0-3.6%, depending on the solar zenith angle and the SeaWiFS geometry. The major source of error is in the calibration of the ground-based radiometer operated in radiance mode, assumed to be accurate to +/- 2%. The establishment of strict criteria for atmospheric stability, angular geometry, and surface conditions resulted in selection of only 26 days for the analysis during 1999-2000 (Venice site) and 1998-2001 (Lanai site). For these days the measured level-1B radiance from the SeaWiFS Project Office was generally lower than the corresponding simulated radiance in band 8 by 7.0% on average, +/- 2.8%. PMID- 16381536 TI - Measuring the optical parameters of weakly absorbing, highly turbid suspensions by a new technique: photoacoustic detection of scattered light. AB - We present and apply a novel method, the scattering photoacoustic (SPA) technique, for measuring optical parameters in weakly absorbing, highly scattering suspensions. In this method, a solid absorber is in contact with a suspension sample to permit the photoacoustic detection of the sample's light scattering properties. We conducted measurements conducted to determine the reduced scattering coefficients of Intralipid suspensions with a concentration range of 0.1-5%, and the results are in good agreement with those achieved by other researchers. Moreover, we also illustrate the relationship between the amplitude of the SPA signal and absorption, scattering, and detection distance. Through a study of Intralipid-ink mixes, we demonstrate that the SPA technique has the ability to determine simultaneously the absorption and reduced scattering coefficients of turbid media. This new technique has low cost and is noninvasive, and it enables on-line measurements to be made. PMID- 16381537 TI - Optical waveguiding in suspensions of dielectric particles. AB - An optical waveguide formed by a suspension of dielectric nanoparticles in a microchannel is described. The suspensions, chosen for their guiding and scattering properties, are silica and polystyrene particles that have diameters of 30-900 nm and are dispersed in water with volume fractions up to 10%. Changing the diameter and concentration of the particles causes the suspensions to transition from Rayleigh to Mie scattering and from single to multiple scattering. The threshold for optical guiding in a waveguide core composed of these suspensions is set by the numerical aperture of the effective refractive index difference introduced by the suspension and not by the average interparticle distance. PMID- 16381538 TI - Multiple-scattering effects on static light-scattering optical structure factor measurements. AB - We show that the extent and effect of multiple scattering on angularly resolved light-scattering intensity measurements, the optical structure factor, can be quantitatively described by a single parameter, the average number of scattering events along the scattering volume. This quantity is easily measured or calculated and hence provides a useful experimental indicator of multiple scattering, which is a hindrance to accurate structure factor measurements. PMID- 16381539 TI - Complete characterization of optical pulses by real-time spectral interferometry. AB - We demonstrate a simple method for complete characterization (of amplitudes and phases) of short optical pulses, using only a dispersive delay line and an oscilloscope. The technique is based on using a dispersive delay line to stretch the pulses and recording the temporal interference of two delayed replicas of the pulse train. Then, by transforming the time domain interference measurements to spectral interferometry, the spectral intensity and phase of the input pulses are reconstructed, using a Fourier-transform algorithm. In the experimental demonstration, mode-locked fiber laser pulses with durations of approximately 1 ps were characterized with a conventional fast photodetector and an oscilloscope. PMID- 16381540 TI - Signal degradation due to output filtering of self-seeded gain-switched pulses exhibiting weak inherent side-mode-suppression ratios. AB - We show the importance of achieving an acceptable level of output side-mode suppression ratio when generating pulses by using the self-seeded gain-switched technique. Experiments carried out on such pulses exhibiting poor side-mode suppression ratios that are subsequently filtered to improve the latter demonstrate that they possess an associated level of noise. This buildup of noise with a decreasing inherent side-mode-suppression ratio is noted regardless of the improved output-filtered side-mode-suppression ratio of 35 dB that is maintained. The degradation of the signal is due to the mode partition effect and may render these pulses unsuitable for use in high-speed optical communications systems. PMID- 16381541 TI - The status of peer review. PMID- 16381542 TI - Hypertension in the oncology setting. AB - In the oncology setting, HTN is a common comorbidity and complication of some treatments. A thorough assessment of each patient, including medical history and current medication list, is used to screen for HTN. The Joint National Committee guidelines (National Heart, Lung, and Blood Institute, 2003) have provided a resource for proper selection of medications for management of HTN, with considerations for special populations. PMID- 16381543 TI - Clinical update: a nonhealing fractured mandible. AB - Bisphosphonates have shown significant clinical benefit in reducing skeletal fractures in patients with multiple myeloma or bone: penicillin VK 500 mg or amoxicillin 500 mg; both 4 times daily (QID) initially and twice daily (BID) for maintenance, If penicillin allergic: 1. Clindamycin 150 to 300 mg QID. 2. Vibramycin 100 mg once daily (QD). 3. Erythromycin ethylsuccinate 400 mg 3 times daily (TID). 4. Antifungals when required: 5. Nystatin oral suspension 5 to 15 mL QID or 100,000 IU/mL. 6. Mycelex troches (clotrimazole 10 mg) x 5/day. 7. Fluconazole 200 mg initially, then 100 mg QD. 8. Other potential systemic antifungals include itraconazole or ketoconazole. 9. Antivirals, if required: 10. Acyclovir 400 mg BID. 11. Valacyclovir hydrochloride 500 mg to 2g BID. PMID- 16381544 TI - Breast cancer education for Native American women: creating culturally relevant communications. AB - In the Navajo language, the word for cancer translates as the sore that does not heal. This literal linkage to a sense of hopelessness reflects a cultural perspective that impedes cancer detection in its early, more treatable stages. As a coauthor of this article and a Navajo breast cancer survivor, Nellie Sandoval, BS, MS, explains that the very topic of cancer is taboo to discuss among the Navajo population, for to speak of cancer is to invite it. When statistical data from the San Juan Regional Tumor Registry supported the authors' anecdotal findings regarding late diagnoses, they created Breast Cancer: It Can Be Healed. The first Navajo-language video to address such cultural barriers, it discusses the triad of early detection-breast self-examination, clinical examination, and mammography. Its success sparked creation of a second video, sponsored by the Native American Cancer Research Partnership (NACRP). The 12-minute video, Breast Cancer: The Healing Begins, focuses on treatment options, including surgery, radiation, and hormone therapy. By conducting field screenings throughout the Navajo Nation, the NACRP team has enhanced the video's visual imagery and messages and has confirmed the value of cultural relevancy in cancer education. PMID- 16381545 TI - Sexual dysfunction related to the treatment of young women with breast cancer. AB - Women have a 13.4% chance of developing breast cancer over their lifetime, and an estimated 60,000 women aged 20-50 will develop breast cancer in 2005. Surgical, radiologic, and chemotherapeutic treatments used to treat breast cancer can alter women's body image and menopausal status, ultimately affecting sexual health. A diagnosis of breast cancer is more traumatic for young women because of psychosocial concerns, side effects of treatment, and a potentially shortened life expectancy. Alterations in sexual health interfere with intimate relationships. Nurses can play a pivotal role in improving the sexual health of young women with breast cancer. This article explores the potential side effects of treatment that can affect sexual health, the unique needs of young women, and nursing interventions that systematically address sexual health concerns. PMID- 16381546 TI - Management of opioid-induced sedation in patients with cancer. AB - Opioid-induced sedation is a common dose-limiting side effect of opioid therapy that can be very distressing and sometimes is more difficult to manage than pain. Opioid-induced sedation may prohibit patients from participating in certain activities of daily living, which can be a source of considerable distress for patients and their families. The issue presents a therapeutic dilemma for healthcare professionals caring for patients experiencing the side effect. Currently, several therapeutic approaches are used to counteract or minimize the severity of opioid-induced sedation, including reduction of the opioid dose, the addition of other drugs, opioid rotation, and the use of invasive routes of administration. This article will address the management of opioid-induced sedation. PMID- 16381547 TI - The phenomenon of chemo brain. AB - During and following chemotherapy, some patients experience difficulties with memory, attention, and other aspects of cognitive function. This constellation of deficits commonly is referred to as chemo brain. Although the phenomenon is not understood completely, it is assuming greater significance as cancer survival improves. Return to prediagnosis levels of domestic, employment, and academic activity is expected in most survivors. Advances in basic, imaging, and clinical sciences are beginning to unravel pathophysiologic mechanisms and develop neuroprotective strategies. Pharmacologic options are borrowed from diverse diseases, including attention-deficit/hyperactivity disorder and neurodegenerative diseases. Conventional therapies soon may find new applications; for example, recent preclinical data suggest that erythropoietin may have some neuroprotective abilities, which may positively affect patients experiencing chemo brain. A collaborative model is bringing together international specialists interested in unraveling the mysteries of the phenomenon and developing management strategies to attenuate its effects. This article will review the clinical features of chemo brain as well as a working hypothesis regarding pathophysiology. The potential and emerging interventions that can be used by oncology nurses to assist patients and their families to cope with this enigmatic dysfunction will be discussed. PMID- 16381548 TI - Shared decision making: empowering the bedside nurse. AB - Shared decision making is a process that has empowered specialty nurses at the Mayo Clinic in Rochester, MN, to solve a practice concern. Staff nurses recognized a lack of concise, collated information available that described what nurses need to know when caring for patients receiving chemotherapy. Many aspects of the administration process were knowledge and experience based and not easily retrievable. The Hematology/Oncology/Blood and Marrow Transplant Clinical Practice Committee identified this as a significant practice issue. Ideas were brainstormed regarding how to make the information available to nursing colleagues. The Chemotherapy Yellow Pages is a resource that was developed to facilitate the rapid retrieval of pertinent information for bedside nurses. The content of this article outlines a'model of shared decision making and the processes used to address and resolve the practice concern. PMID- 16381549 TI - Hospice patients in clinical cancer treatment trials. AB - Patients with cancer and practitioners face a conflict when no known curative treatments exist because Medicare and other insurers do not allow clinical trial participation and hospice care to occur concurrently. Patients may not fully benefit from hospice care because of late enrollment. Barriers to hospice referrals exist in the form of clinician attitudes and insurance limitations. Patients with advanced cancer may be prevented from enrolling in clinical trials, and trial participation may limit hospice care. Factors related to this situation are discussed. PMID- 16381550 TI - Understanding bodywork for the patient with cancer. AB - The goal of this article is to increase oncology nurses' understanding of common bodywork modalities and the current research about them in the oncology setting. Bodywork is a broad term that incorporates massage and energy modalities. Eleven modalities are described. In addition, issues related to safety, licensure, making referrals, and nurses' and bodyworkers' roles are discussed. Better knowledge will increase oncology nurses' abilities to assess and guide patients' bodywork choices and facilitate discussions with patients, physicians, and bodyworkers to ensure that patients with cancer are receiving safe and effective care. PMID- 16381552 TI - Editorial 2006: exciting and challenging times for organic letters. PMID- 16381551 TI - Review of selected approaches to promoting smoking cessation. PMID- 16381553 TI - Two polymerizable derivatives of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). AB - [structure: see text] A versatile strategy for the synthesis of polymerizable derivatives of the redox-active indicator dye 2,2'-azino-bis(3 ethylbenzthiazoline-6-sulfonic acid) (ABTS) is reported. Two products are shown to illustrate how the final step in the synthetic strategy can be used to alter the physical properties of the product. Both products were characterized spectroscopically and electrochemically. The hydrophilic monomer (sABTS) was polymerized, and the utility of this polymer (polyABTS) is demonstrated in the context of bioelectrocatalysis. PMID- 16381554 TI - Amphidinolide B: asymmetric synthesis of a C7-C20 synthon. AB - [reaction: see text] An asymmetric synthesis of a C(7)-C(20) synthon of amphidinolide B is described. The synthesis entails the construction of C(7) C(13) and C(14)-C(20) fragments and makes extensive use of catalytic asymmetric bond constructions to establish the requisite stereochemical relationships. Fragment coupling proceeds by Suzuki cross-coupling and installs the trisubstituted diene unit that is among amphidinolide B's defining structural features. PMID- 16381555 TI - Microwave-enhanced palladium-catalyzed cross-coupling reactions of potassium vinyltrifluoroborates and allyl acetates: a new route to 1,4-pentadienes. AB - [reaction: see text] A novel and straightforward microwave synthesis of 1,4 pentadienes has been developed involving the cross-coupling of potassium vinyltrifluoroborates and allyl acetates in the presence of a palladium catalyst. PMID- 16381556 TI - Asymmetric synthesis of propargylamides via 3,3'-disubstituted binaphthol modified alkynylboronates. AB - [reaction: see text] Alkynylboronates derived from 3,3'-disubstituted-2,2' binaphthols react with various N-acylimines to give the expected chiral propargylamides with up to 99% ee. This new methodology was applied to the first enantioselective synthesis of the antitubulin agent (-)-N-acetylcolchinol. PMID- 16381557 TI - A convenient FeCl3-catalyzed hydroarylation of styrenes. AB - [reaction: see text] 1,1-Diarylalkanes are easily synthesized by CH functionalization reactions of electron-rich arenes and heteroarenes with styrenes in the presence of FeCl(3) as catalyst. PMID- 16381558 TI - Synthetic studies toward the haouamines. AB - [reaction: see text] A concise synthetic approach toward the haouamines based on Stork-Danheiser alkylation and Friedel-Crafts chemistry is described. A novel electrophilic aromatic substitution with concomitant formation of an enol triflate is reported. PMID- 16381559 TI - Stereoselective furan-iminium cation cyclization in the construction of the core structure of manzamine A. AB - [reaction: see text] A new type of furan-iminium cation cyclization was developed and used to construct the ABC ring of manzamine A. The cyclization proceeded at the 2-position with complete regio- and stereoselectivity to give a spiro-center. The product was efficiently converted to the highly substituted core structure of manzamine A. PMID- 16381560 TI - Induction of gamma-turn-like structure in ferrocene bearing dipeptide chains via conformational control. AB - [structure: see text] A combination of the ferrocene scaffold as a central reverse-turn unit with the dipeptide chains (-L-Pro-L-Ala-NHPy) was demonstrated to induce both inverse gamma-turn-like and antiparallel beta-sheet-like structures. Only the antiparallel beta-sheet-like structure was formed in the ferrocene bearing the heterochiral dipeptide chains (-L-Pro-D-Ala-NHPy), in which highly organized self-assembly was achieved through a network of intermolecular hydrogen bonds. PMID- 16381561 TI - Computational characterization of a complete palladium-catalyzed cross-coupling process: the associative transmetalation in the Stille reaction. AB - [reaction: see text] The species presumably involved in the associative ligand substitution mechanism for the Stille cross coupling of vinyl bromide and trimethylvinyl stannane with Pd(PMe(3))(2)/PMe(3) as catalysts in DMF (as ligand and solvent) have been structurally and energetically characterized. The cyclic four-coordinate transition state for the rate-determining transmetalation step explains the retention of configuration in the Stille coupling of chiral nonracemic alkyl stannanes. PMID- 16381562 TI - Solvent-dependent changes in the ene reaction of RTAD with alkenes: the cyclopropyl group as a mechanistic probe. AB - [reaction: see text] The vinylcyclopropyl moiety was used as an efficient probe to test mechanistic possibilities of the triazolinedione-alkene ene reaction. In non-hydroxylic solvents, this reaction afforded only the ene adducts via a closed three-membered aziridinium imide (AI) intermediate, whereas in hydroxylic solvents a dipolar intermediate is favored and trapped by the cyclopropyl moiety to form the corresponding cyclopropyl-rearranged solvent-trapped adducts. PMID- 16381563 TI - Addition of arylboronic acids to symmetrical and unsymmetrical azo compounds. AB - [reaction: see text] The addition of aryl- and heteroarylboronic acids to azo compounds is described. Copper salt catalysis was necessary to perform the reaction under mild conditions and high yields. Excellent regioselectivity was observed in addition to unsymmetrical azo compounds. PMID- 16381564 TI - Investigation of the Yamaguchi esterification mechanism. Synthesis of a lux-s enzyme inhibitor using an improved esterification method. AB - [reaction: see text] A one-pot procedure for the regioselective synthesis of aliphatic esters is described. This was a result of a study on mixed aliphatic aromatic anhydrides. The data suggest that during the Yamaguchi esterification reaction, a symmetric aliphatic anhydride is produced in situ, which upon reaction with an alcohol yields the ester. We confirmed that benzoyl chloride could be used instead of the sterically hindered Yamaguchi acid chloride. This method was successfully applied in the synthesis of Lux-S aspartic acid inhibitor. PMID- 16381565 TI - Stereoselective hydroazidation of amino enones: synthesis of the ritonavir/lopinavir core. AB - [reaction: see text] The base-catalyzed hydroazidation of alpha'-amino alpha,beta unsaturated ketones with in situ generated hydrazoic acid was found to proceed with high stereoselectivity in favor of the syn product. The stereoselectivity is controlled by the configuration of the enone and syn/anti ratios up to 7:1 were obtained with secondary and tertiary amines at low temperature. By this route the diamino alcohol core of HIV-PR inhibitors ritonavir and lopinavir was synthesized in 37% yield from phenylalanine. PMID- 16381566 TI - Synthesis of the ribosomal P-site substrate CCA-pcb. AB - [reaction: see text] CCA-pcb (cytidylyl-(3'5')-cytidylyl-(3'5')-3'(2')-O-(N-(6-D (+)-biotinoylaminohexanoyl)-L-phenylalanyl)adenosine), a ribosomal P-site substrate, was synthesized by phosphoramidite chemistry in 26 steps with an overall yield of 18%, starting from biotin. The synthesis relies on the judicious selection of orthogonal silyl protecting groups for the 5'-hydroxyls and acid labile protecting groups (DMTr, AcE, and MeE) at other reactive sites to ensure the intactness of the labile ester. Both 3'-esterification and nucleotide coupling were accomplished by in situ activation with imidazolium ions. PMID- 16381567 TI - Preparation of 2,5,7-trithiabicyclo[2.2.1]heptane from 1,2-bis triisopropylsilanylsulfanyl alkenes. AB - [reaction: see text] The chemical behavior of 1,2-bis-triisopropylsilanylsulfanyl alkenes 1 is relatively unexplored, and the weak sulfur-silicon bonds give rise to various transformations. Under acidic conditions (HCl) and in the presence of a Lewis acid at room temperature the bicyclic adduct 2 is obtained in good yield. The structure was confirmed by X-ray crystal analysis with R = benzyl. PMID- 16381568 TI - Vinylation of aryl bromides using an inexpensive vinylpolysiloxane. AB - [reaction: see text] A mild and general method for the palladium-catalyzed vinylation of aryl bromides has been developed. The use of tetrabutylammonium fluoride (TBAF) as the activator and an inexpensive and nontoxic vinyl donor, 1,3,5,7-tetramethyl-1,3,5,7-tetravinylcyclotetrasiloxane (D(4)(V), 1), allows for a general and high-yielding preparation of substituted styrenes. PMID- 16381569 TI - Rhodium-catalyzed addition of alkynes to activated ketones and aldehydes. AB - [reaction: see text] The rhodium-catalyzed addition of alkynes to 1,2-diketones, 1,2-ketoesters, and aldehydes provides a method for the synthesis of tertiary alkynyl alcohols under mild conditions. The reaction tolerates many functional groups (such as carboxylic acids) that are incompatible with other methods. The alkyne addition reaction proceeds best using bulky phosphine ligands such as 2 (di-tert-butylphosphino)biphenyl. This method fills a void in the more common zinc-catalyzed processes, which give poor yields with enolizable 1,2-dicarbonyl substrates. PMID- 16381570 TI - 2-(1,3-Dioxan-2-yl)ethylsulfonyl group: a new versatile protecting and activating group for amine synthesis. AB - [reaction: see text] 2-(1,3-Dioxan-2-yl)ethylsulfonyl (Dios) chloride was synthesized and used as a new versatile sulfonating agent for amines. Primary and secondary amines were sulfonated very easily in excellent yields with Dios chloride. N-Nonsubstituted and N-monosubstituted Dios-amides, activated amines, were alkylated satisfactorily under new Mitsunobu conditions utilizing (cyanomethylene)tributylphosphorane (CMBP). The Dios group is very stable under basic and reductive conditions and is removed by heating in a hot aqueous solution of trifluoroacetic acid. PMID- 16381571 TI - cis-dihydroxylation of unsaturated potassium alkyl- and aryltrifluoroborates. AB - [reaction: see text] The cis-dihydroxylation of olefin-containing potassium alkyl and aryltrifluoroborates proceeds readily in moderate to excellent yields. The resulting diols are efficient coupling partners in Suzuki-Miyaura-type reactions with both alkenyl and aryl bromides. PMID- 16381572 TI - Configurational analysis of cyclopropyl fatty acids isolated from Escherichia coli. AB - [reaction: see text] The absolute configuration of methyl lactobacillate and its 9,10 homologue, both isolated from Escherichia coli B-ATCC 11303, was found to be 11R,12S and 9R,10S, respectively. PMID- 16381573 TI - Co2(CO)8-catalyzed intramolecular hetero-Pauson-Khand reaction of alkynecarbodiimide: synthesis of (+/-)-physostigmine. AB - [reaction: see text] Herein we describe a novel Co(2)(CO)(8)-catalyzed intramolecular aza-Pauson-Khand-type reaction of alkynecarbodiimide derivatives affords pyrrolo[2,3-b]indol-2-one ring systems in reasonable yields. This is the first reported Co(2)(CO)(8) successfully applied in the hetero-Pauson-Khand reaction. Significantly, the transformation of one of our pyrrolo[2,3-b]indol-2 one derivatives into the indole alkaloid, (+/-)-physostigmine, was completed in a highly stereoselective manner. PMID- 16381574 TI - Tin-free intermolecular addition of primary alkyls to imines via the dimethylzinc air radical process. AB - [reaction: see text] A dimethylzinc-air initiator was applied to the generation of primary alkyl radicals from alkyl iodides. The addition of the generated primary alkyl radicals to N-tosylimines was accelerated by the action of boron trifluoride-diethyl etherate and copper(II) triflate to give the corresponding adducts in good yields after 2-3 h. Air oxygen was essential for the reaction to proceed, showing involvement of a radical process in the reaction. PMID- 16381575 TI - Synthesis and properties of a dithiirane trans-1,2-dioxide, a three-membered vic disulfoxide. AB - cis- and trans-3-(1-adamantyl)-3-tert-butyldithiirane 1-oxides were oxidized with dimethyldioxirane to give the trans-1,2-dioxide. Thermal decomposition of the 1,2 dioxide yielded the corresponding (E)- and (Z)-sulfines, thioketone, and cis- and trans-dithiirane 1-oxides. In the thermolysis, decomposition to the sulfines and SO was the main path and that to the thioketone and SO(2) was the minor one. The two decomposition processes and epimerization to the cis-1,2-dioxide were analyzed theoretically. SO generated in situ reacted with thioketones as additives to give the corresponding dithiirane 1-oxides. PMID- 16381576 TI - Sequential pericyclic reaction of ene-diallenes: an efficient approach to the steroid skeleton. AB - [reaction: see text] The one-pot construction of polycyclic aromatic systems from acyclic ene-bis(propargyl alcohols) was achieved through a tandem dual [2,3] sigmatropic rearrangement/6pi-electrocyclic reaction/intramolecular [4 + 2] cycloaddition sequence. A steroidal compound was conveniently synthesized using the present method. PMID- 16381577 TI - Synthesis and photochromism of novel phenylene-linked photochromic bispyrans. AB - [reaction: see text] Phenylene-linked bisnaphthopyrans were synthesized in good yields via the one-pot reaction of bis-propargyl alcohols with naphthols. Temperature-dependent photochromism in 1,4-phenylene-linked bispyrans leads to up to 60 nm bathochromic shift between the colored species formed at room temperature and at -20 degrees C. Better fatigue resistance and higher colorability was observed in 1,4-phenylene-linked bis-[2H]-naphtho[1,2-b]pyrans by comparison to the 1,3-phenylene linked bis-[2H]-naphtho[1,2-b]pyrans. PMID- 16381578 TI - Phosphorinanes as ligands for palladium-catalyzed cross-coupling chemistry. AB - [structure: see text] Phosphorinanes are presented as a class of phosphine ligand suitable for organopalladium cross-coupling chemistry. Prepared via a direct double Michael addition of a monoalkyl- or arylphosphine to phorone followed by a Wolf-Kishner reduction, phosphorinanes are relatively inexpensive to manufacture and allow modification of one of the alkyl moieties permitting steric and electronic fine-tuning of the ligands. Library screening and applications of these ligands in the Suzuki, Sonogashira, ketone arylation, and aryl amination reactions are presented. PMID- 16381579 TI - Application of RCM reaction in the construction of ABC ring of micrandilactone A. AB - [reaction: see text] The functionalized ABC ring system of micrandilactone A was successfully constructed in 14 steps. The key reactions in this synthesis are the intermolecular Diels-Alder reaction (IMDA) and the eneyne ring-closing metathesis (RCM) reaction. PMID- 16381580 TI - Synthesis of new cholesterol- and sugar-anchored squaraine dyes: further evidence of how electronic factors influence dye formation. AB - [reaction: see text] Synthesis of new quinaldine-based squaraine dyes linked to cellular recognition elements that exhibit near-infrared absorption (>740 nm) are described. Both product analysis and theoretical calculations substantiate the interesting electronic effects of various substituents in the dye formation reaction. These results are useful in the synthesis of symmetrical and unsymmetrical squaraine dyes that can have potential biological and photodynamic therapeutical applications. PMID- 16381582 TI - The alpha-effect in gas-phase SN2 reactions revisited. AB - This paper re-examines gas-phase S(N)2 reactions at saturated carbon for model reactions Nu(-) + CH(3)Cl --> CH(3)Nu + Cl(-) (Nu(-) = HO(-), MeO(-), NH(2)(-), HS(-), Cl(-), Br(-), I(-), HOO(-), MeOO(-), HSS(-), and NH(2)NH(-)) using the G2(+) theory. The calculated results show that the alpha-effect does exist in the gas-phase S(N)2 reaction at the sp(3) carbon, contrary to the currently accepted notion of the absence of the alpha-effect in the gas phase. PMID- 16381581 TI - Facile construction of the pentacyclic framework of subincanadine B. Synthesis of 20-deethylenylated subincanadine B and 19,20-dihydrosubincanadine B. AB - [reaction: see text] We describe a facile approach for effectively constructing the pentacyclic framework of subincanadine B. The seven-step assembly of tetracyclic ketone 14 featured Michael addition, Pictet-Spengler cyclization, and Dieckmann condensation. From this key ketone intermediate, two analogues of subincanadine B, i.e., 20-deethylenylated subincanadine B (27) and 19,20 dihydrosubincanadine B (31), were synthesized in four steps, respectively. PMID- 16381583 TI - Lycoperine A, A novel C27N3-type pentacyclic alkaloid from Lycopodium hamiltonii, inhibiting acetylcholinesterase. AB - [structure: see text] A novel C(27)N(3)-type pentacyclic Lycopodium alkaloid, lycoperine A (1) consisting of two octahydroquinoline rings and a piperidine ring, was isolated from the club moss Lycopodium hamiltonii. The structure and relative stereochemistry were elucidated on the basis of 2D NMR data and chemical transformation. Lycoperine A (1) exhibited an inhibitory activity against acetylcholinesterase. PMID- 16381584 TI - Enantioselective synthesis of vicinal halohydrins via dynamic kinetic resolution. AB - [reaction: see text] Expanding the scope of enantioselective catalysis via DKR, transfer hydrogenation of a variety of cyclic alpha-halo ketones was accomplished using the Noyori/Ikariya (R,R)- or (S,S)-I catalysts and either HCO(2)H/Et(3)N or HCO(2)Na/n-Bu(4)NBr in H(2)O/CH(2)Cl(2) as the hydrogen sources. Good yields of vicinal bromo-, chloro-, and fluorohydrins with excellent de and ee levels were achieved in most cases after a simple tuning of reaction conditions. PMID- 16381585 TI - An alpha-formylglycine building block for fmoc-based solid-phase peptide synthesis. AB - [reaction: see text] Nearly all known sulfatases share a common active site modification that is required for their activity: conversion of cysteine to alpha formylglycine. We report the synthesis of an alpha-formylglycine building block suitable for Fmoc-based solid-phase peptide synthesis. The building block was incorporated into a synthetic peptide derived from the active site of a Mycobacterium tuberculosis sulfatase. PMID- 16381586 TI - Synthesis of the N-acetylcysteamine thioester of seco-proansamitocin. AB - [structure: see text] The enantioselective total synthesis of the N acetylcysteamine thioester of seco-proansamitocin, a key biosynthetic intermediate of the highly potent antitumor agent ansamitocin, is described, which twice utilizes the Nagao acetate aldol reaction, as well as an indium mediated alkynylation of a benzyl bromide followed by carboalumination. The key step is a Heck reaction between two terminal alkenes for merging the two major fragments. PMID- 16381587 TI - Selective diethylzinc reduction of imines in the presence of ketones catalyzed by Ni(acac)2. AB - A selective reduction method of an electronically deficient imine in the presence of ketone was developed by employing Et(2)Zn and 5 mol % of Ni(acac)(2). The method was applied in the reduction of S(S)-tert-butanesulfinyl ketimines 1 to afford amines 2 in 23-92% yields and 73:27 to 98:2 diastereoselectivities. A plausible mechanism was proposed on the basis of an NMR study. PMID- 16381588 TI - Enantioselective addition of terminal alkynes to isolated isoquinoline iminiums. AB - [reaction: see text] The asymmetric, catalytic addition of terminal alkynes to discrete alkyliminium species in the presence of copper bromide, QUINAP ligand, and triethylamine is reported. The reaction proceeds in high yield and high enantiomeric excess with a variety of substrates in solution and is also amenable to solid-phase synthesis. PMID- 16381589 TI - Cyano(ethoxycarbonothioylthio)methyl benzoate: a novel one-carbon radical equivalent. AB - [reaction: see text] Cyano(ethoxycarbonothioylthio)methyl benzoate 3 has been prepared and shown to be an excellent one-carbon radical equivalent that can be applied for the introduction of an acyl unit via xanthate transfer radical addition to olefins. The corresponding adducts can be further elaborated. A rare 1,5-nitrile translocation was also observed during the study. PMID- 16381590 TI - An enantioselective total synthesis of (+)-ricciocarpin A. AB - [reaction: see text] Starting from 4,4-dimethyl-2-cyclohexenone, an efficient total synthesis of ricciocarpin A (1) in natural form has been accomplished. PMID- 16381591 TI - Synthesis of the tetracyclic core of the kempanes by a ring-closing metathesis strategy. AB - [reaction: see text] The synthesis of the tetracyclic ring system of the kempane diterpenes was achieved through the highly regio- and stereoselective Diels-Alder reaction of an isopropenyl-diene with 2,6-dimethyl-p-benzoquinone, addition of an allyl group, and ring-closing metathesis of the isopropenyl and allyl groups. PMID- 16381592 TI - Convergent, stereoselective synthesis of the GHIJ fragment of brevetoxin A. AB - [reaction: see text] A stereoselective synthesis of the GHIJ fragment of brevetoxin A utilizing a convergent assembly strategy is described. Glycolate alkylation, ring-closing metathesis, and Hosomi-Sakurai reactions were central operations in the construction of the G ring and J ring subunits, which were united through a Horner-Wadsworth-Emmons coupling. Subsequent dehydrative cyclization produced an endocyclic enol ether that was further elaborated to the tetracyclic GHIJ fragment of brevetoxin A. PMID- 16381593 TI - The first examples of Nazarov cyclizations leading to annulated pyrroles. AB - [reaction: see text] Reactions between alpha-substituted unsaturated carboxylic acids 20 and N-tosylpyrroles [14, 23] in the presence of trifluoroacetic anhydride result in smooth alpha-acylation of the pyrrole, followed by Nazarov cyclization to give 50-80% yields of cyclopenta[b]pyrroles. The presence of an alpha-substituent in the unsaturated acid appears to be mandatory. PMID- 16381594 TI - Facile preparation and reactivity of polymer-supported N-(2-Iodyl-phenyl) acylamide, an efficient oxidizing system. AB - A simple three-step preparation of polymer-supported N-(2-iodyl-phenyl)-acylamide (NIPA resin) starting from 2-iodoaniline is described. The resin was obtained with good loading levels (0.7-0.8 mmol g(-1)) and has been successfully used for efficient oxidation of a diverse collection of alcohols. Thus, treating alcohols with 1.0 equiv of the resin in 1,2-dichloroethane under reflux for 30-60 min allowed rapid and in most cases complete conversion to the corresponding carbonyl compound. PMID- 16381595 TI - Theory-guided discovery of unique chemical transformations of cyclopropenes. AB - [reaction: see text] Chiral 2-cyclopropenyl-4-tolyl sulfones, available by the [2 + 1]-cycloaddition of tosyldiazomethane to acetylenes under catalysis by the Rh(II) complex 1, provide a number of unusual transformations and useful chiral products. This chemistry is dominated by the unusual strain and reactivity of the cyclopropene ring system. PMID- 16381596 TI - Claisen-type condensation of vinylogous acyl triflates. AB - [reaction: see text] The Claisen-type condensation reaction of cyclic vinylogous carboxylic acid triflates with lithium enolates and their analogues produces acyclic alkynes bearing a 1,3-diketone-type moiety. The present transformation is proposed to proceed via a 1,2-addition of the enolate to the vinylogous acyl triflate, followed by fragmentation of the aldolate intermediate. PMID- 16381597 TI - An alternative and effective HIV vaccination approach based on inhibition of antigen presentation attenuators in dendritic cells. AB - BACKGROUND: Current efforts to develop HIV vaccines that seek to stimulate immune responses have been disappointing, underscoring the inability of natural immune responses to control HIV-1 infection. Here we tested an alternative strategy to induce anti-HIV immune responses by inhibiting a host's natural immune inhibitor. METHODS AND FINDINGS: We used small interfering RNA (siRNA) to inhibit suppressor of cytokine signaling (SOCS) 1, a key negative regulator of the JAK/STAT pathway, and investigated the effect of this silencing on the ability of dendritic cells (DCs) to induce anti-HIV-1 immunity. We found that SOCS1-silenced DCs broadly induced enhanced HIV-1 envelope (Env)-specific CD8+ cytotoxic T lymphocytes and CD4+ T helper cells, as well as antibody responses, in mice. Importantly, SOCS1 silenced DCs were more resistant to HIV Env-mediated suppression and were capable of inducing memory HIV Env-specific antibody and T cell responses. SOCS1 restricted signaling, as well as production of proinflammatory cytokines such as interleukin-12 by DCs, play a critical role in regulating the anti-HIV immune response. Furthermore, the potency of HIV DNA vaccination is significantly enhanced by coimmunization with SOCS1 siRNA expressor DNA. CONCLUSIONS: This study demonstrates that SOCS1 functions as an antigen presentation attenuator to control both HIV-1-specific humoral and cellular responses. This study represents the first, to our knowledge, attempt to elicit HIV-specific T cell and antibody responses by inhibiting a host's antigen presentation attenuator, which may open a new and alternative avenue to develop effective therapeutic and prophylactic HIV vaccines. PMID- 16381598 TI - Cost-effectiveness of alternative blood-screening strategies for West Nile Virus in the United States. AB - BACKGROUND: West Nile virus (WNV) is endemic in the US, varying seasonally and by geographic region. WNV can be transmitted by blood transfusion, and mandatory screening of blood for WNV was recently introduced throughout the US. Guidelines for selecting cost-effective strategies for screening blood for WNV do not exist. METHODS AND FINDINGS: We conducted a cost-effectiveness analysis for screening blood for WNV using a computer-based mathematical model, and using data from prospective studies, retrospective studies, and published literature. For three geographic areas with varying WNV-transmission intensity and length of transmission season, the model was used to estimate lifetime costs, quality adjusted life expectancy, and incremental cost-effectiveness ratios associated with alternative screening strategies in a target population of blood-transfusion recipients. We compared the status quo (baseline screening using a donor questionnaire) to several strategies which differed by nucleic acid testing of either pooled or individual samples, universal versus targeted screening of donations designated for immunocompromised patients, and seasonal versus year long screening. In low-transmission areas with short WNV seasons, screening by questionnaire alone was the most cost-effective strategy. In areas with high levels of WNV transmission, seasonal screening of individual samples and restricting screening to blood donations designated for immunocompromised recipients was the most cost-effective strategy. Seasonal screening of the entire recipient pool added minimal clinical benefit, with incremental cost effectiveness ratios exceeding USD 1.7 million per quality-adjusted life-year gained. Year-round screening offered no additional benefit compared to seasonal screening in any of the transmission settings. CONCLUSIONS: In areas with high levels of WNV transmission, seasonal screening of individual samples and restricting screening to blood donations designated for immunocompromised recipients is cost saving. In areas with low levels of infection, a status-quo strategy using a standard questionnaire is cost-effective. PMID- 16381599 TI - Aspartic proteinase content of the Arabidopsis genome. AB - The sequence of the Arabidopsis genome has given us information about one plant's complement of aspartic proteinases. Using an in silico analysis based on the homology to known aspartic proteinase genes, we have uncovered 51 sequences that potentially encode these enzymes. This is substantial more than the number predicted for other eukaryotic systems. We have grouped the deduced amino acid sequences into 3 classes - typical plant aspartic proteinase, nucellin-like and atypical aspartic proteinase sequences-, depending on their putative domain organizations and their active site sequence motifs. Searching databases has revealed cDNAs or ESTs for nearly 90% of these genes. Sequence analysis using software that detects targeting signals indicates most of the predicted proteins have the expected localization in the secretory system although several of these are membrane bound. The analysis also predicts 8 chloroplast localized proteins and 2 mitochondria-localized aspartic proteinase-like proteins. The wide variety of structures and subcellular locations implies multiple functions for aspartic proteinases in plants. PMID- 16381600 TI - Mapping of the active site of proteases in the 1960s and rational design of inhibitors/drugs in the 1990s. AB - For several decades the specificity of proteases has been presented as an active site divided into subsites, using the nomenclature of Schechter & Berger from 1967 (S1, S2... for subsites of the active site; P1, P2... for residues of the substrate occupying the corresponding subsites). At early stages of the research (1960s) it was realized that the size of the active site was larger than expected and important interactions occur in regions remote from the catalytic site. Since the active site was found to be large it was divided into subsites, and a procedure to map it up was developed. The map provides information on the size of the active site (number of subsites), the properties of each subsite (free energy of ligand binding, nature of binding forces, etc.), and it enables rational design of new substrates and inhibitors. Already in 1968 inhibitors with binding constants ten thousand fold higher than available inhibitors, were prepared. The model of a large active site was initially met with strong opposition. Before long, however, predictions of the model (size of the active site, interactions in subsites remote from the catalytic site) were confirmed by X-ray crystallography (1970). During the 1990s proteolytic enzymes received renewed attention in biology and medicine, they became therapeutic targets, and protease inhibitors were successfully applied in the treatment of AIDS and hypertension. The model of large active site divided into subsites, proposed 38 years ago, stood the test of time. This model is still in use in basic research to evaluate enzyme activity, and in pharmaceutical research for the development of inhibitors/drugs. PMID- 16381601 TI - Nepenthesin, a unique member of a novel subfamily of aspartic proteinases: enzymatic and structural characteristics. AB - Carnivorous plants are known to secrete acid proteinases to digest prey, mainly insects, for nitrogen uptake. In our recent study, we have purified, for the first time, to homogeneity two acid proteinases, nepenthesin I (Nep I) and nepenthesin II (Nep II) from the pitcher fluid of Nepenthes distillatoria and investigated their enzymatic and structural characteristics. Both enzymes were optimally active at pH approx. 2.6 toward acid-denatured hemoglobin; the specificity of Nep I toward oxidized insulin B chain appears to be similar, but slightly wider than those of other aspartic proteinases (APs). At or below 50 degrees C both enzymes were remarkably stable; especially Nep I was extremely stable over a wide range of pH from 3 to 10 for over 30 days. This suggests an evolutionary adaptation of the enzymes to their specific habitat. We have also cloned the cDNAs and deduced the complete amino acid sequences of the precursors of Nep I and Nep II from the pitcher tissue of Nepenthes gracilis. Although the corresponding mature enzymes are homologous with ordinary pepsin-type APs, both enzymes had a high content of cysteine residues (12 residues per molecule), which are assumed to form six unique disulfide bonds as suggested by computer modeling and are thought to contribute toward the remarkable stability of Neps. Moreover, the amino acid sequence identity of Neps with ordinary APs, including plant vacuolar APs, are remarkably low (approx. 20%), and phylogenetic comparison shows that Neps are distantly related to them to form a novel subfamily of APs with a high content of cysteine residues and a characteristic insertion, named 'the Nep type AP (NAP)-specific insertion', including a large number of novel, orthologous plant APs emerging in the gene/protein databases. PMID- 16381602 TI - Computational methods for remote homolog identification. AB - As more and more protein sequences are available, homolog identification becomes increasingly important for functional, structural, and evolutional studies of proteins. Many homologous proteins were separated a very long time ago in their evolutionary history and thus their sequences share low sequence identity. These remote homologs have become a research focus in bioinformatics over the past decade, and some significant advances have been achieved. In this paper, we provide a comprehensive review on computational techniques used in remote homolog identification based on different methods, including sequence-sequence comparison, and sequence-structure comparison, and structure-structure comparison. Other miscellaneous approaches are also summarized. Pointers to the online resources of these methods and their related databases are provided. Comparisons among different methods in terms of their technical approaches, their strengths, and limitations are followed. Studies on proteins in SARS-CoV are shown as an example for remote homolog identification application. PMID- 16381603 TI - Endothelin receptors in gastrointestinal smooth muscle. AB - Endothelins (ETs) are a family of peptides with 21-amino-acid residues. ET-1 was identified as a potent vasoconstrictor produced by vascular endothelial cells. Three distinct isoforms of ET, i.e. ET-1, ET-2 and ET-3, have been found to exist in a variety of tissues. ET was later found to cause contraction as well as relaxation of smooth muscle in many physiologic systems. In the gastrointestinal tract, ET causes contraction and/or relaxation of the esophagus, stomach, ileum and colon. In the hepatobiliary system, ET causes contraction of the portal vein, hepatic stellate cells, gallbladder and common bile duct. In mammalian species, two classes of ET receptors, ET(A) and ET(B), have been cloned. ET(A) receptors have higher affinities for ET-1 and ET-2 than ET-3, while ET(B) receptors have the same affinities for ET-1, ET-2 and ET-3. In the gastrointestinal system, ET causes smooth muscle contraction through interaction with ET(A) receptors, ET(B) receptors or both ET(A) and ET(B) receptors, depending on the tissues and species. In addition to contraction, ET causes smooth muscle relaxation through interaction with ET(A) receptors or ET(B) receptors. At the present time, there are no studies showing that ET causes smooth muscle relaxation through interaction with both ET(A) and ET(B) subtypes. ET induces contraction in most of the non-sphincter muscle except the fundus of the stomach. On the other hand, ET causes relaxation and contraction in the lower esophageal and internal anal sphincters. ET may play an important role in the control of human gastrointestinal motility and portal vein pressure. PMID- 16381604 TI - Naturally occurring polyamines: interaction with macromolecules. AB - The naturally occurring polyamines, spermine [NH2(CH2)3NH(CH2)4NH(CH2)3NH2] and spermidine [NH2(CH2)3NH(CH2)4NH2], as well as the diamine putrescine [NH2(CH2)4NH2], are widely spread in nature. They occur in plants, micro organisms and animal tissues and fulfil many important physiological functions. Due to their cationic nature they interact with negatively charged macromolecules such nucleic acids, phospholipids and proteins. This ionic interaction, which is reversible, leads to the stabilization of DNA, tRNA, membranes and some proteins. Early studies demonstrated that polyamines stimulate the growth of pro- and eukaryotic cells and that they play an important role in carcinogenesis and in malignant transformation processes. As a result of these studies various inhibitors of polyamine biosynthesis have been synthesized and are used to combat cancer and parasitic diseases (e.g., African sleeping sickness). PMID- 16381605 TI - The RING-finger protein haprin: domains and function in the acrosome reaction. AB - The RBCC (RING finger, B-box type zinc finger, coiled-coil domain) motif family contains a large number of proteins implicated in many cellular processes, including vesicle exocytosis. The acrosome reaction, the sperm exocytotic event that is required for fertilization, involves essentially the same process of intracellular membrane fusions as vesicular exocytosis in somatic cells. We have previously isolated a haploid-germ-cell-specific gene designated haprin, which encodes a RBCC motif protein that plays a role in the acrosome reaction of sperm by mediating protein complex formation via the RBCC motif. In this review, we describe the potential role of Haprin in the molecular mechanisms of acrosome reaction, as compared with some other RBCC proteins. The conserved structure and localization of the Haprin protein in human and mouse suggest an indispensable role for Haprin in the functioning of mammalian sperm. PMID- 16381606 TI - IX International Conference on Salt Lake research: research opportunities and management challenges. AB - The 9th International Conference on Salt Lake Research was held 26-30 September 2005 in Western Australia at the Curtin University of Technology, Perth, Australia. One hundred scientists from 10 countries presented research on a diverse array of topics highlighting research findings and opportunities, and management challenges associated with inland saline waters. Major emergent themes of the conference included modeling of ecosystem processes, microbial communities, and features of Western Australian inland saline environments, including current threats, conservation and management. PMID- 16381607 TI - Greater risk of incident asthma cases in adults with allergic rhinitis and effect of allergen immunotherapy: a retrospective cohort study. AB - Asthma and rhinitis are often co-morbid conditions. As rhinitis often precedes asthma it is possible that effective treatment of allergic rhinitis may reduce asthma progression. The aim of our study is to investigate history of allergic rhinitis as a risk factor for asthma and the potential effect of allergen immunotherapy in attenuating the incidence of asthma. Hospital-referred non asthmatic adults, aged 18-40 years between 1990 and 1991, were retrospectively followed up until January and April 2000. At the end of follow up, available subjects were clinically examined for asthma diagnosis and history of allergen specific immunotherapy, second-hand smoking and the presence of pets in the household. A total of 436 non-asthmatic adults (332 subjects with allergic rhinitis and 104 with no allergic rhinitis nor history of atopy) were available for final analyses. The highest OR (odds ratio) associated with a diagnosis of asthma at the end of follow-up was for the diagnosis of allergic rhinitis at baseline (OR, 7.8; 95%CI, 3.1-20.0 in the model containing the covariates of rhinitis diagnosis, sex, second-hand smoke exposure, presence of pets at home, family history of allergic disorders, sensitization to Parietaria judaica; grass pollen; house dust mites; Olea europea: orchard; perennial rye; and cat allergens). Female sex, sensitization to Parietaria judaica and the presence of pets in the home were also significantly predictive of new onset asthma in the same model. Treatment with allergen immunotherapy was significantly and inversely related to the development of new onset asthma (OR, 0.53; 95%CI, 0.32-0.86). In the present study we found that allergic rhinitis is an important independent risk factor for asthma. Moreover, treatment with allergen immunotherapy lowers the risk of the development of new asthma cases in adults with allergic rhinitis. PMID- 16381608 TI - Comparison of normalization methods for CodeLink Bioarray data. AB - BACKGROUND: The quality of microarray data can seriously affect the accuracy of downstream analyses. In order to reduce variability and enhance signal reproducibility in these data, many normalization methods have been proposed and evaluated, most of which are for data obtained from cDNA microarrays and Affymetrix GeneChips. CodeLink Bioarrays are a newly emerged, single-color oligonucleotide microarray platform. To date, there are no reported studies that evaluate normalization methods for CodeLink Bioarrays. RESULTS: We compared five existing normalization approaches, in terms of both noise reduction and signal retention: Median (suggested by the manufacturer), CyclicLoess, Quantile, Iset, and Qspline. These methods were applied to two real datasets (a time course dataset and a lung disease-related dataset) generated by CodeLink Bioarrays and were assessed using multiple statistical significance tests. Compared to Median, CyclicLoess and Qspline exhibit a significant and the most consistent improvement in reduction of variability and retention of signal. CyclicLoess appears to retain more signal than Qspline. Quantile reduces more variability than Median in both datasets, yet fails to consistently retain more signal in the time course dataset. Iset does not improve over Median in either noise reduction or signal enhancement in the time course dataset. CONCLUSION: Median is insufficient either to reduce variability or to retain signal effectively for CodeLink Bioarray data. CyclicLoess is a more suitable approach for normalizing these data. CyclicLoess also seems to be the most effective method among the five different normalization strategies examined. PMID- 16381609 TI - Changes in microRNA expression profiles in HIV-1-transfected human cells. AB - MicroRNAs (miRNAs) are small RNAs of 18-25 nucleotides (nt) in length that play important roles in regulating a variety of biological processes. Recent studies suggest that cellular miRNAs may serve to control the replication of viruses in cells. If such is the case, viruses might be expected to evolve the ability to modulate the expression of cellular miRNAs. To ask if expression of HIV-1 genes changes the miRNA profiles in human cells, we employed a high throughput microarray method, termed the RNA-primed Array-based Klenow Enzyme (RAKE) assay. Here, we describe the optimization of this assay to quantify the expression of miRNAs in HIV-1 transfected human cells. We report distinct differences in miRNA profiles in mock-transfected HeLa cells versus HeLa cells transfected with an infectious HIV-1 molecular clone, pNL4-3. PMID- 16381610 TI - Functional studies of signaling pathways in peri-implantation development of the mouse embryo by RNAi. AB - BACKGROUND: Studies of gene function in the mouse have relied mainly on gene targeting via homologous recombination. However, this approach is difficult to apply in specific windows of time, and to simultaneously knock-down multiple genes. Here we report an efficient method for dsRNA-mediated gene silencing in late cleavage-stage mouse embryos that permits examination of phenotypes at post implantation stages. RESULTS: We show that introduction of Bmp4 dsRNA into intact blastocysts by electroporation recapitulates the genetic Bmp4 null phenotype at gastrulation. It also reveals a novel role for Bmp4 in the regulation the anterior visceral endoderm specific gene expression and its positioning. We also show that RNAi can be used to simultaneously target several genes. When applied to the three murine isoforms of Dishevelled, it leads to earlier defects than previously observed in double knock-outs. These include severe delays in post implantation development and defects in the anterior midline and neural folds at headfold stages. CONCLUSION: Our results indicate that the BMP4 signalling pathway contributes to the development of the anterior visceral endoderm, and reveal an early functional redundancy between the products of the murine Dishevelled genes. The proposed approach constitutes a powerful tool to screen the functions of genes that govern the development of the mouse embryo. PMID- 16381611 TI - The Academic Medical Center Linear Disability Score (ALDS) item bank: item response theory analysis in a mixed patient population. AB - BACKGROUND: Currently, there is a lot of interest in the flexible framework offered by item banks for measuring patient relevant outcomes. However, there are few item banks, which have been developed to quantify functional status, as expressed by the ability to perform activities of daily life. This paper examines the measurement properties of the Academic Medical Center linear disability score item bank in a mixed population. METHODS: This paper uses item response theory to analyse data on 115 of 170 items from a total of 1002 respondents. These were: 551 (55%) residents of supported housing, residential care or nursing homes; 235 (23%) patients with chronic pain; 127 (13%) inpatients on a neurology ward following a stroke; and 89 (9%) patients suffering from Parkinson's disease. RESULTS: Of the 170 items, 115 were judged to be clinically relevant. Of these 115 items, 77 were retained in the item bank following the item response theory analysis. Of the 38 items that were excluded from the item bank, 24 had either been presented to fewer than 200 respondents or had fewer than 10% or more than 90% of responses in the category 'can carry out'. A further 11 items had different measurement properties for younger and older or for male and female respondents. Finally, 3 items were excluded because the item response theory model did not fit the data. CONCLUSION: The Academic Medical Center linear disability score item bank has promising measurement characteristics for the mixed patient population described in this paper. Further studies will be needed to examine the measurement properties of the item bank in other populations. PMID- 16381613 TI - Novel oxygen-concentrator-based equipment: take a test drive first! PMID- 16381612 TI - Classification of real and pseudo microRNA precursors using local structure sequence features and support vector machine. AB - BACKGROUND: MicroRNAs (miRNAs) are a group of short (approximately 22 nt) non coding RNAs that play important regulatory roles. MiRNA precursors (pre-miRNAs) are characterized by their hairpin structures. However, a large amount of similar hairpins can be folded in many genomes. Almost all current methods for computational prediction of miRNAs use comparative genomic approaches to identify putative pre-miRNAs from candidate hairpins. Ab initio method for distinguishing pre-miRNAs from sequence segments with pre-miRNA-like hairpin structures is lacking. Being able to classify real vs. pseudo pre-miRNAs is important both for understanding of the nature of miRNAs and for developing ab initio prediction methods that can discovery new miRNAs without known homology. RESULTS: A set of novel features of local contiguous structure-sequence information is proposed for distinguishing the hairpins of real pre-miRNAs and pseudo pre-miRNAs. Support vector machine (SVM) is applied on these features to classify real vs. pseudo pre miRNAs, achieving about 90% accuracy on human data. Remarkably, the SVM classifier built on human data can correctly identify up to 90% of the pre-miRNAs from other species, including plants and virus, without utilizing any comparative genomics information. CONCLUSION: The local structure-sequence features reflect discriminative and conserved characteristics of miRNAs, and the successful ab initio classification of real and pseudo pre-miRNAs opens a new approach for discovering new miRNAs. PMID- 16381614 TI - A prospective comparison of 3 new-generation pulse oximetry devices during ambulation after open heart surgery. AB - OBJECTIVE: To assess the clinical performance of 3 new-generation pulse-oximetry signal-processing software systems (Philips FAST, Masimo SET, and Nellcor N-3000) during ambulation after open-heart surgery. DESIGN: Prospective, convenience sample. SETTING: Cardiac surgical progressive care unit in a 629-bed, not-for profit, tertiary-care teaching hospital. PATIENTS: Status post-cardiac-surgery patients (n = 36) during their first postoperative ambulation. INTERVENTIONS: None. PROTOCOL: Randomization was used for digit and hand selection, and all 3 devices were used continuously during ambulation. Data on dropouts, false alarms, and correlation with heart rate were recorded. We continuously measured arterial oxygen saturation via pulse oximetry during ambulation with all 3 devices. RESULTS: Pairwise comparisons indicated significant differences among the 3 devices for data dropout and false alarm. In repeated-measures analysis, the Nellcor N-3000 had the greatest likelihood of data dropout (odds ratio of 31.9 to Masimo and 5.6 to Philips, at the 95% confidence interval). However, the converse was true for false alarms; the Masimo had the most false alarms, with an odds ratio of 17.9 to Nellcor and 2.3 to Philips, at the 95% confidence interval. There were also significantly more dropouts with all 3 devices when readings were taken on a hand on an arm from which a radial graft had been taken (p = 0.004). For heart-rate correlation, the mean absolute difference among the 3 devices was similar: Philips = 4.3 beats/min, Masimo = 5.1 beats/min, and Nellcor = 3.0 beats/min. CONCLUSIONS: There are significant differences among the 3 devices with regard to dropout and false alarms. High numbers of dropouts are problematic because no pulse-oximetry patient information is available during dropout. However, false alarms are even more problematic, because they desensitize clinicians to alarms and call into question the accuracy of displayed data. While our data highlight the statistical differences between the studied oximeters, the clinical implications of these differences warrant further study. PMID- 16381615 TI - The effect of telephone appointment-reminder calls on outpatient absenteeism in a pulmonary function laboratory. AB - BACKGROUND: Absenteeism from outpatient appointments is common. Telephone appointment-reminder calls reduce outpatient-appointment absenteeism in many clinic settings. OBJECTIVE: To determine if telephone appointment-reminder calls reduce outpatient absenteeism at a hospital-based pulmonary function laboratory. METHODS: We conducted a retrospective review of our pulmonary function laboratory's outpatient appointment records from April to November 2004. Data were collected from consecutive outpatient appointments, including patient age, sex, whether a telephone appointment-reminder call was successfully made, and whether the patient showed up for the scheduled test. We performed 3 analyses. Differences in absenteeism between the groups was the primary outcome measure. First, appointments were separated into 2 groups: (1) appointments for which a reminder call was attempted ("called" group) and (2) appointments for which a reminder call was not attempted ("not-called" group). The appointments were then separated into 2 further groups: (1) the reminder call was successfully achieved ("contacted" group) and (2) the patient either was not called or was called but could not be reached ("not-contacted" group). Finally, the contacted group was separated into 2 further groups: (1) reminder calls that resulted in direct conversation with an appropriate person at the patient's listed telephone number, and (2) reminder message left on an answering machine. RESULTS: Data were collected from 515 consecutive outpatient appointments; 45 (8.7%) of these patients did not show up for testing. The absentee rate was 4.7% (n = 10) in the called group and 11.6% (n = 35) in the not-called group (p = 0.0066). In the called group, 6.5% (n = 14) could not be reached. The absentee rate was 4% (n = 8) in the contacted group and 11.7% (n = 37) in the not-contacted group (p = 0.0021). We found no difference in absenteeism between patients who received reminders via direct conversation (4.2%) and those who had a reminder message left on an answering machine (3.7%) (p > 0.05). CONCLUSIONS: A policy of reminding outpatients of their appointments via telephone reduces absenteeism at a hospital-based pulmonary function laboratory. We found no difference in absenteeism between communicating the reminder via direct conversation versus via leaving a message on an answering machine. PMID- 16381616 TI - A proposed curvilinearity index for quantifying airflow obstruction. AB - BACKGROUND: Though forced expiratory volume in the first second (FEV(1)) is the primary indicator of airway obstruction, curvilinearity in the expiratory flow volume curve is used to support the quantitative assessment of obstruction via FEV(1). Currently there is no available index to quantify a pathological contour of curvilinearity. STUDY PURPOSE: We propose a "curvature" index (k(max)) and compare FEV(1) values to the index with a sequential sample of spirometry data. METHODS: The hyperbolic function b(0)Q + b(1)Q V + b(2)V = 1 (in which Q = flow rate, V = volume, and b(0), b(1), and b(2) are estimated from the patient's flow volume data) is fit to a fixed segment of the descending phase of the expiratory flow-volume curve. A previously developed biomechanical interpretation of this relationship associates the coefficient b(1) with the rate of airway-resistance increase as exhaled volume increases. A global curvature index k(max)=b(1)/2(b(0)b(2)+b(1)) is defined to quantify the curvilinearity phenomenon. We used statistics software to determine the k(max) of spirometry data from 67 sequential patients, and to determine the relationship of k(max) to FEV(1). RESULTS: Individual k(max) estimates appeared to correspond well with the degree of curvilinearity observed and were related in an exponential manner to FEV(1). CONCLUSIONS: We defined a curvature index to quantify the curvilinearity phenomenon observed in the expiratory limb of flow-volume loops from patients with obstructive lung disease. This index uses data from a major segment of the flow-volume curve, and our preliminary data indicate an exponential relationship with FEV(1). This new index allows the putative association between curvilinearity and obstructive lung disease to be examined quantitatively in clinical practice and future studies. PMID- 16381617 TI - A comparison of 2 methods of continuous aerosol administration during methacholine challenge testing. AB - BACKGROUND: Exposure to the bronchoconstricting agent methacholine is a potential hazard to technical staff during methacholine challenge testing, which remains a useful and frequently performed test. There are several methods of performing the test. One of the 2 methods listed in the American Thoracic Society's guidelines is the 2-min tidal-breathing method. The methacholine can be inhaled using one of several methods. The loosely-fitting-mask method is likely to produce more contamination of the local environment than a filtered exhalation system. METHODS: We tested 2 variations of the tidal-breathing method of measuring the methacholine provocational concentration (PC(20), the dose that produces a 20% decrease in forced expiratory volume in the first second). One involved use of the open-mask technique and the other a T-piece-and-filter system that precluded the release of methacholine-containing droplets into the environment. We performed duplicate methacholine challenge tests with 10 subjects who had a wide range of PC(20). The tests were done in random order, and each subject performed one test using the mask and one using the T-piece/filter system. RESULTS: With the mask system the geometric mean PC(20) was 4.7 mg/mL, versus 5.1 mg/mL with the T-piece-filter system (p = 0.36). These values are very close and would not be substantially different clinically. CONCLUSION: The 2 methods are equivalent, and the low cost of the products used in the T-piece/filter method makes it suitable for reducing technician exposure to methacholine, using potentially completely disposable components. PMID- 16381618 TI - Asthma educational seminar targeting Medicaid providers. AB - BACKGROUND: Medicaid-insured children have high risk of asthma but are less likely to receive care in keeping with national guidelines. We targeted providers who care for a large proportion of Medicaid-insured children and presented a 2 session multimedia asthma-education seminar that emphasizes communication and teaching techniques, to enhance providers' asthma-care teaching skills. METHODS: Five Medicaid-approved health maintenance organizations recruited pediatric primary-care providers. Providers were surveyed at baseline, 6 months, and 12 months to determine if they reported changes in their use of certain asthma-care communication techniques. RESULTS: Fifty-three of 70 participating providers completed the program and initial survey. They reported that 50% (median) of their patients were insured by Medicaid. At baseline, providers reported they were very confident of their asthma knowledge; however, they were less confident in interactions with patients/families regarding asthma self-management skills. Providers reported use of written plans less than half of the time. The response rate was 60% at 6 months and 71% at 1 year. Twenty-eight providers completed all 3 surveys. They reported significantly more frequent use of communication and counseling techniques that involved patient/parent asthma education and self management skills at the 6-month point, that were partly sustained at 1 year. Reported provision of written asthma plans to patients had increased significantly at 6 months, but that increase was not sustained at the 1-year point. CONCLUSIONS: The seminar significantly enhanced knowledge of specific communication techniques related to asthma-teaching goals and reported use of asthma action plans to enhance self-management skills; however, these practices appear to need frequent reinforcement. PMID- 16381619 TI - An investigation of nebulized bronchodilator delivery using a pediatric lung model of spontaneous breathing. AB - BACKGROUND: The literature lacks comparative data on nebulizer aerosol delivered via mask versus T-piece, to spontaneously breathing pediatric subjects. PURPOSE: To compare total inhaled drug mass delivered via standard pediatric aerosol mask versus via T-piece, with increasing distance. METHODS: We used a sample of 5 nebulizers, operated under manufacturers' conditions, with a standard pediatric aerosol mask and with a T-piece capped at one end, at 0 cm, 1 cm, and 2 cm from an inhalation filter placed at the inlet of a pediatric test lung. Inhaled drug mass was analyzed with spectrophotometry. Aerosol particle size was measured separately from the breathing simulations, using a laser particle sizer to determine fine-particle mass (particles < 4.7 mum) and fine-particle fraction as percent of total mass. The fine-particle fraction was used to estimate the fine particle mass. RESULTS: The mean + SD values for inhaled drug mass as a percentage of nominal dose, at 0 cm, 1 cm, and 2 cm, with the mask were 2.88 + 0.79%, 1.61 + 0.65%, and 1.3 + 0.42%, respectively, and with the T-piece were 4.14 + 1.37%, 3.77 + 1.04%, and 3.47 + 0.64%, respectively. There was a statistically greater inhaled drug mass with T-piece than with mask, overall (p < 0.01), and a significant decrease with mask or T-piece as distance increased (p < 0.01). The difference between mask and T-piece for inhaled drug mass at 2 cm was statistically significant (p < 0.018). The mean + SD values for fine-particle mass estimated as a percentage of total drug mass at 0, 1, and 2 cm, with the mask were 1.39 + 0.36%, 0.78 + 0.29%, and 0.64 + 0.20%, respectively, and with the T-piece were 2.1 + 0.63%, 1.84 + 0.45%, and 1.71 + 0.27%, respectively. CONCLUSION: Inhaled drug mass was greater with T-piece than with a standard pediatric aerosol mask under the conditions studied. PMID- 16381620 TI - Conflicting methacholine challenge tests. PMID- 16381621 TI - Highlights From ECCO 13-The European Cancer Conference Paris, France October 30 to November 3, 2005. PMID- 16381622 TI - The global breast cancer burden: variations in epidemiology and survival. AB - Breast cancer is the most common type of cancer and the most common cause of cancer-related mortality among women worldwide. However, the burden is not evenly distributed, and, according to the best available data, there are large variations in the incidence, mortality, and survival between different countries and regions and within specific regions. Many complex factors underlie these variations, including population structure (eg, age, race, and ethnicity), lifestyle, environment, socioeconomic status, risk factor prevalence, mammography use, disease stage at diagnosis, and access to high-quality care. We review recent breast cancer incidence and mortality statistics and explore why these vary so greatly across the world. Further research is needed to fully understand the reasons for variations in breast cancer outcomes. This will aid the development of tailored strategies to improve outcomes in general as well as the standard of care for underserved populations and reduce the burden of breast cancer worldwide. PMID- 16381623 TI - Concepts and clinical trials of dose-dense chemotherapy for breast cancer . AB - This article will review the strategy of dose-dense administration of chemotherapy for breast cancer. Increased dose density is achieved by reducing the interval between each dose of chemotherapy. The cumulative drug dose remains constant, but the same amount of drug is administered over a shorter period. Mathematical models of tumor growth have provided the basis for the clinical application of dose-dense chemotherapy. The Norton-Simon model suggests that increasing the dose density of chemotherapy will increase efficacy by minimizing the opportunity for regrowth of tumor cells between cycles of chemotherapy. Intergroup trial 9741, coordinated by the Cancer and Leukemia Group B (CALGB), tested the 2 hypotheses that dose-dense and sequential administration of chemotherapy regimens incorporating doxorubicin, cyclophosphamide, and paclitaxel would improve disease-free survival and overall survival. A statistically significant 4-year disease-free survival advantage was detected for the 2 dose dense regimens compared with the regimens administered every 3 weeks. The mathematical concepts and previous clinical trials of dose density that contributed to the design of CALGB 9741 will be reviewed. The strengths and limitations of CALGB 9741 will then be discussed before the presentation of future directions of research and recommendations for clinical practice today. PMID- 16381624 TI - Reducing the global breast cancer burden: the importance of patterns of care research. AB - Breast cancer treatment guidelines are not uniformly followed in clinical practice, with evidence for substantial variations in treatment patterns, quality of care, and patient outcomes among and within countries. The factors that drive treatment decisions are unclear. Furthermore, the impact of different treatment strategies on survival is poorly understood outside the clinical trial setting. Sources of patterns of care information often have limitations in completeness, quality, timeliness of reporting, and relevance to the larger population. Patterns of care studies frequently lack details on cancer stage at diagnosis, tumor biology, and treatment received. It is difficult to compare data between studies and/or track changes over time because of variations in data sources and collection techniques. Thus, the design and implementation of a global registry is sorely needed in order to prospectively evaluate worldwide patterns of care and outcomes in patients with breast cancer. Components of this registry should include random selection of centers of variable practice settings in multiple countries and accurate and rapid data reporting at prestudy and follow-up timepoints. Data collected would include tumor and demographic factors, staging information, treatment rendered, and survival. Variables that influenced the treatment selected would be assessed. This unique international effort would allow the development of strategies to improve diagnostic and treatment-related standards of care and survival outcomes, thus reducing the breast cancer burden worldwide. PMID- 16381625 TI - Chemotherapy-induced amenorrhea from adjuvant breast cancer treatment: the effect of the addition of taxanes. AB - PURPOSE: Adjuvant chemotherapy for breast cancer can be associated with a variety of side effects, one of which is the induction of premature menopause in premenopausal patient. Although taxanes have increasingly been used in the adjuvant setting, there has been relatively little published on the frequency of amenorrhea related to their use. PATIENTS AND METHODS: We review records of 159 premenopausal patients receiving adjuvant chemotherapy from our practice. RESULTS: Altogether, 51% of all patients retained menstrual function after chemotherapy. CONCLUSION: It was observed that patients receiving adjuvant anthracycline-based chemotherapy with sequential taxane therapy did not have a higher rate of amenorrhea than those not receiving a taxane. PMID- 16381626 TI - Two concurrent phase II trials of paclitaxel/carboplatin/trastuzumab (weekly or every-3-week schedule) as first-line therapy in women with HER2-overexpressing metastatic breast cancer: NCCTG study 983252. AB - PURPOSE: The efficacy and tolerability of 2 different schedules of paclitaxel/carboplatin/trastuzumab for HER2-overexpressing metastatic breast cancer (MBC) were evaluated in this parallel multicenter phase II trial. PATIENTS AND METHODS: Patients received every-3-week therapy (n = 43) consisting of a 200 mg/m(2) dose of paclitaxel/carboplatin area under the curve (AUC) of 6 mg/mL per minute and trastuzumab (an initial 8 mg/kg dose and subsequent 6 mg/kg doses) administered every 21 days for 8 cycles or weekly therapy (n = 48) consisting of an 80-mg/m(2) dose of paclitaxel/carboplatin AUC of 2 mg/mL per minute for 3 of 4 weeks, with weekly trastuzumab (an initial 4-mg/kg dose and subsequent 2-mg/kg doses) administered every 4 weeks for 6 cycles. Trastuzumab was continued until disease progression or unacceptable toxicity. HER2 status was confirmed by a central laboratory review. RESULTS: The overall response rate (ORR) with every-3 week therapy was 65% (90% confidence interval [CI], 51%-77%), with a median time to disease progression of 9.9 months and median overall survival (OS) time of 2.3 years. The ORR with weekly therapy was 81% (90% CI, 70%-90%), with a median time to disease progression of 13.8 months and a median OS time of 3.2 years. Hematologic and nonhematologic toxicities occurred significantly less frequently with weekly therapy versus every-3-week therapy: grade 3/4 neutropenia (52% vs. 88%); grade 3 thrombocytopenia (4% vs. 30%); and grade 3 neurosensory toxicity (2% vs. 19%), respectively. CONCLUSIONS: Every-3-week and weekly regimens of paclitaxel/carboplatin/trastuzumab are highly active in women with HER2 overexpressing MBC. However, fewer patients developed severe neutropenia, leukopenia, or thrombocytopenia with the weekly schedule. PMID- 16381627 TI - Docetaxel/gemcitabine administered every other week as first-line treatment for metastatic breast cancer: final results of a phase II trial. AB - BACKGROUND: The purpose of this study was to assess the toxicity and efficacy of the combination of docetaxel and gemcitabine every 2 weeks as first-line therapy in metastatic breast cancer. PATIENTS AND METHODS: We selected patients between the ages of 18 years and 70 years with a Karnofsky performance status of > or = 60 to receive docetaxel 65 mg/m(2) followed by gemcitabine 2500 mg/m(2) on day 1 every 14 days for 10 cycles. Patients could receive more cycles at the discretion of investigator. RESULTS: Of the 52 patients entered, 48 were evaluable for efficacy and toxicity. The overall response rate was 75% (95% confidence interval, 60%-86%), with 8 patients (17%) exhibiting a complete response. The median time to progression was 10.7 months, and the median survival was 32.2 months. The predominant grade 3/4 hematologic toxicity was neutropenia (44% of patients), and the predominant grade 3/4 nonhematologic toxicity was asthenia (15% of patients). Other grade 3/4 toxicities, such as anemia, nausea/vomiting, and diarrhea, were present but infrequent (< or = 10% of patients). The relative dose intensities of both drugs were > 88%. CONCLUSION: The combination of docetaxel/gemcitabine once every 2 weeks is active and well tolerated in patients with untreated advanced breast carcinoma. This chemotherapy regimen might be useful in advanced breast cancer when anthracycline combinations are not preferred, such as cases previously treated with adjuvant anthracycline chemotherapy. PMID- 16381628 TI - Dermal lymphatic emboli in inflammatory and noninflammatory breast cancer: a French-Tunisian joint study in 337 patients. AB - BACKGROUND: We studied whether dermal lymphatic emboli (DLE) add independent prognostic information to the clinical definition of inflammatory breast cancer (IBC). PATIENTS AND METHODS: The study was performed in 2 centers, one each in France and Tunisia. For every patient with IBC, 1-3 patients with noninflammatory breast cancer (non-IBC) were included. All patients were to have a surgical tumor biopsy, including a sample of the skin surrounding the tumor. The endpoint was the risk of a relapse at 2 years, which was estimated using univariate and multivariate Cox models. RESULTS: Three hundred thirty-seven patients were included (150 in France and 187 in Tunisia). The IBC status was divided into 2 clinical categories according to the extent of inflammation in the breast (localized IBC, which was defined as clinical inflammation in the tumor area, vs. diffuse IBC, which was defined as inflammation of at least two thirds of the breast). In total, 57 patients presented with localized IBC, 71 with diffuse IBC, and 209 with non-IBC. Dermal lymphatic emboli were found in 7% of non-IBC cases, in 25% of localized IBC cases, and in 45% of diffuse IBC cases. We found a significant interaction between the presence of DLE and diffuse IBC (P = 0.01). In patients with diffuse IBC, the presence of DLE increased the risk of relapse 3 fold. Conversely, DLE were not associated with the risk of relapse in patients with non-IBC, nor in patients with localized IBC. In patients with diffuse IBC and no DLE, the risk of relapse was similar to that of patients with localized IBC. CONCLUSION: A DLE status might be a useful prognostic indicator exclusively in patients with diffuse IBC. However, because all patients with localized and diffuse IBC generally receive similar types of treatment, additional information on the presence or absence of DLE will not have an impact on treatment practice. PMID- 16381629 TI - Treatment of chemotherapy-induced anemia in breast cancer: results of a randomized controlled trial of darbepoetin alfa 200 microg every 2 weeks versus epoetin alfa 40,000 U weekly. AB - BACKGROUND: Current chemotherapy regimens for breast cancer result in high incidences of anemia, which can be treated with erythropoietic agents. The relative efficacy of darbepoetin alfa and epoetin alfa was explored in this phase II, open-label, randomized, multicenter trial in anemic patients with breast cancer receiving chemotherapy. PATIENTS AND METHODS: Patients were randomized at a 1:1 ratio to receive darbepoetin alfa 200 microg every 2 weeks (n = 72) or epoetin alfa 40,000 U weekly (n = 69) for < or = 16 weeks. Clinical and hematologic endpoints and validation of a novel patient satisfaction questionnaire for anemia treatment were evaluated for all patients randomized to receive > or = 1 dose of study drug. RESULTS: Baseline characteristics were generally similar between treatment groups. Mean changes in hemoglobin (Hb) level from baseline were similar at 1.9 g/dL for darbepoetin alfa and 1.7 g/dL for epoetin alfa. Hematopoietic responses (> or = 2 g/dL increase in Hb level from baseline or Hb level > or = 12 g/dL) were also similar between groups (88% for darbepoetin alfa and 81% for epoetin alfa). The proportions of patients who received a transfusion during treatment were 6% (95% CI, 0-11%) for darbepoetin alfa and 16% (95% CI, 7%-25%) for epoetin alfa. Most patients (67 patients receiving darbepoetin alfa [93%]; 61 patients receiving epoetin alfa [90%]) exhibited a clinically meaningful target Hb level > or = 11 g/dL. No differences in safety were observed. CONCLUSION: These results suggest that, in patients with breast cancer, darbepoetin alfa 200 microg every 2 weeks and epoetin alfa 40,000 U weekly result in comparable clinical outcomes for the treatment of chemotherapy induced anemia. PMID- 16381630 TI - Statins: health-promoting agents show promise for breast cancer prevention. PMID- 16381631 TI - A pilot study to evaluate the vascular endothelial growth factor receptor tyrosine kinase inhibitor AZD2171 and chemotherapy in locally advanced and inflammatory breast cancer. PMID- 16381632 TI - [An experimental study on antiviral effects of IFN alpha]. AB - OBJECTIVE: To investigate the effects of different subtypes IFN alpha (IFN alpha2b, IFN alpha2a, and IFN alpha1b) transduction molecular STAT1, STAT2, IFNAR, PKR, and RNase L, and to study the differences of their antiviral effects and to evaluate the key signaling transduction molecules. METHODS: (1) After HepG2 cells were treated with IFN alpha2b, IFN alpha2a, or IFN alpha1b, the mRNA levels of STAT1, STAT2, IFNAR, PKR, and RNase L were detected by RT-PCR. (2) After HepG2 cells were treated with 1000 U/ml IFN alpha2b, IFN alpha2a, or IFN alpha1b, the protein expression levels of STAT1 and IFNAR were examined by Western blot. RESULTS: RT-PCR results: (1) IFNAR, STAT1, and STAT2 mRNA expression levels were slightly higher in the IFN alpha1b group than those in the IFN alpha2b group (P > 0.05). The mRNA expression levels in IFN alpha1b or IFN alpha2b groups were significantly higher than in the IFN alpha2a group (P < 0.05). (2) The PKR mRNA expression showed no significant differences among IFN alpha1b, IFN alpha2b, and IFN alpha2a groups. (3) The RNase L mRNA expression was very weak. We could not compare the differences of the RNase L mRNA levels in different groups by RT-PCR. Western blot results: (1) The IFNAR, and STAT1 protein expressions were greatly up-regulated after IFN alpha induction compared with the untreated group (P < 0.05). (2) The IFNAR, and STAT1 protein expression levels in IFN alpha1b group were slightly higher than the IFN alpha2b group. IFNAR, and STAT1 protein levels of IFN alpha1b or IFN alpha2b group were significantly higher than IFN alpha2a group (P < 0.05). CONCLUSION: STAT1, STAT2, IFNAR mRNA and protein expressions could all be markedly up-regulated after IFN alpha treatment. Effects of IFN alpha1b or IFN alpha2b were greatly stronger than IFN alpha2a. The PKR mRNA expression also was greatly up-regulated after IFN alpha treatment. Expression levels of PKR in IFN alpha1b, IFN alpha2b, and IFN alpha2a groups were all similar. The mRNA level results were consistent with the protein level results. Our results showed that the antiviral activity of IFN alpha1b or IFN alpha2b were stronger than that of IFN alpha2a. The signal transduction molecules STAT1, STAT2, and IFNAR could be regarded as a key index to evaluate antiviral activity of IFN alpha. Further confirmation is still needed to see whether PKR could be regarded as a key index. PMID- 16381633 TI - [The promoter activity of the DNA sequence corresponding to HCV 5'UTR in HepG2]. AB - OBJECTIVE: To study the promoter activity in HepG2 cells of the DNA sequence corresponding to the HCV 5'UTR. METHODS: Plasmids, 5'UTR-Luc(+) and 5'UTR-Luc(-) carrying the forward and reverse DNA sequences corresponding to the HCV 5'UTR respectively were constructed, and subsequently transfected into HepaG2 cells. The luciferase activity and the mRNA of the luciferase gene were then detected. The 5'UTR sequence was cloned into a GFP vector to make 5'UTR-EGFP, and then the GFP expression was confirmed by fluorescence microscopy. RESULTS: 5'UTR-Luc(+) had an obvious luciferase activity whereas 5'UTR-Luc(-) had nearly no luciferase activity. The former had a high level of luciferase mRNA while the latter could not be detected. An intense green fluorescence expression was observed in the cells transfected with the plasmid of 5'UTR-EGFP. CONCLUSION: The forward DNA sequence corresponding to HCV 5'-UTR had an obvious promoter activity in hepG2 cells. It may play an important role in the replication of HCV. PMID- 16381634 TI - [Cleavage of HCV by HCV specific deoxyribozyme in vitro]. AB - OBJECTIVE: To study the cleavage activity of specific deoxyribozyme to hepatitis C virus in vitro. METHODS: Three deoxyribozymes were designed to cleave at sites 157, 168, 173 in HCV 5'-noncoding region with the active region of 5' GGCTAGCTACAACGA-3' respectively. Plasmid pCMV/T7-NCRC -Delta Luc was completely linearized with restriction endonuclease Xba I. HCV RNA5'-NCRC was transcribed in vitro from the linearized products and radiolabelled with [alpha-32P] UTP. Under the conditions of 37 degrees C, pH7.5, Mg2+ 10 mmol/L, the three deoxyribozymes were mixed with substrate RNA individually for 120 minutes and then the reactions were terminated. The cleavaged products were separated with 8% denaturated polyacrylamide gel electrophoresis and displayed by autoradiography. DRz3 was mixed with the substrate RNA at different Mg2+ concentrations. The cleavage efficiency was analyzed with a gel document action analyzing systems. RESULTS: Under the adopted conditions the three deoxyribozymes efficiently cleaved to the target RNA in vitro and the cleavage activity of DRz3 was increased with the increase of Mg2+ concentration. CONCLUSION: The designed deoxyribozymes can cleave 5'-NCR mRNA of HCV efficiently in vitro and it is dose-respondent to Mg2+ concentration. PMID- 16381635 TI - [Effects of nuclear factor kappaB and transforming growth factor beta1 in the anti-liver fibrosis process using Ginkgo biloba extract]. AB - OBJECTIVE: To evaluate the effects of Ginkgo biloba extract (EGB) on CCl(4) induced liver fibrosis and to investigate the underlying mechanisms. METHODS: Rats were divided into the following groups: normal control group, CCl(4) model group, low dose EGB group, moderate dose EGB group and high dose EGB group. The rat liver fibrosis model was induced by intraperitoneal injection of CCl(4) twice a week for 8 weeks. The model rats of the three EGB treated groups were given 0.25 g/kg, 0.5 g/kg, 1.0 g/kg of EGB by stomach tubes every day. At the end of the eighth week, the blood and liver specimens were obtained. The expressions of nuclear factor kappaB (NF-kappaB) P65, and alpha-smooth muscle actin (alpha-SMA) were detected by immunohistochemistry. Radioimmunoassay was exploited to evaluate serum hyaluronic acid (HA) and laminin (LN) levels. Electrophoretic mobility shift assay (EMSA) was used to confirm the nuclear translocation activity of NF kappaB in liver tissues. The mRNA expression of transforming growth factor-beta1 (TGFbeta1) and collagen I was determined by RT-PCR. Malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in liver tissues and alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the sera were also examined. RESULTS: CCl(4) administration induced liver fibrosis, which was inhibited by EGB in a dose-dependent manner. The histopathologic scores of liver fibrosis, the levels of serum ALT, AST, HA and LN were significantly lower in the rats treated with EGB compared with those not treated (P <0.01 or P <0.05). SOD and GSH-Px activities were notably elevated and MDA content was significantly lower in the rats treated with EGB (P <0.01 or P <0.05), indicating reduced oxidative stress. Immunohistochemical staining demonstrated inhibition of hepatic stellate cell (HSC) activation (in terms of alpha-SMA expression) and NF kappaB P65 expression in the livers of the EGB-treated rats. As determined by EMSA and RT-PCR, activation of NF-kappaB, the mRNA expression of TGFbeta1 and collagen I were significantly higher in model group rats, but obviously lower in EGB treated rats. CONCLUSION: EGB is able to ameliorate liver injury and prevent rats from CCl(4)-induced liver fibrosis by suppressing oxidative stress. This process may be related to inhibiting the expression of TGFbeta1 and the induction of NF-kappaB on HSC activation. PMID- 16381636 TI - [The relationship between the plasma homocysteine level and the polymorphism of MTHFR gene C667T in liver cirrhosis]. AB - OBJECTIVE: To study the relationship between the plasma homocysteine (HCY) level and the polymorphism of N(5), N(10)-methylenetetrahydrofolate reductase (MTHFR) gene C667T in liver cirrhosis. METHODS: 112 normal subjects and 87 liver cirrhosis patients were recruited in the study. Their plasma HCY levels were measured using high performance liquid chromatography with fluorescence detection and polymorphisms of their MTHFR gene were analyzed using PCR-RFLP. RESULTS: The mean level of plasma HCY was significantly higher in patients with liver cirrhosis (21.71+/-4.86) micromol/L than that in healthy individuals (8.34+/ 3.59) micromol/L. There were three kinds of MTHFR genotypes: +/+ (TT, homozygous mutation), +/- (CT, heterozygous mutation) and -/- (CC, wild type). The frequencies of the three genotypes were as follows: +/+, 29.9%; +/-, 52.9%; -/-, 17.2% in cirrhosis patients and +/+, 19.6%; +/-, 33.9%; -/-, 46.4% in normal subjects. The frequency of homozygous or heterozygous mutation was significantly higher in cirrhosis patients than that in the normal control. Moreover, plasma homocysteine level was markedly higher in patients with MTHFR genetic mutation than those without mutation. CONCLUSIONS: Hyperhomocysteinemia may be an independent risk factor for liver cirrhosis. MTHFR is the main enzyme related to homocysteine metabolism. The genetic mutation of MTHFR C667T is possibly an important mechanism of hyperhomocysteinemia in liver cirrhosis. The level of plasma homocysteine may be an early indicator for liver cirrhosis. PMID- 16381637 TI - [A quantitative study of the relationship between levels of liver fibrosis markers in sera and fibrosis stages of liver tissues of patients with chronic hepatic diseases]. AB - OBJECTIVES: To study the quantitative relationship between the levels of serum liver fibrosis markers and fibrosis stages of liver tissues in patients with chronic hepatic diseases. METHODS: In 118 patients with chronic hepatitis, fatty liver or cirrhosis, their Serum levels of LN, HA, PCIII and CIV were investigated by EIA and their liver histological changes were studied. The relationship between the levels of serum LN, HA, PCIII and CIV and the degrees of liver tissue fibrosis was analyzed quantitatively by using the SPSS11.0. RESULTS: A correlation between the levels of serum LN, HA, PCIII and CIV and the histologically assessed grades of inflammatory activity was found (r = 0.394, 0.449, 0.443, 0.351, respectively, P <0.01). The correlation between the levels of serum LN, HA, PCIII and CIV and the histological assessed stages of liver fibrosis was strong (r = 0.456, 0.564, 0.476, 0.421 respectively, P <0.01). The levels of serum LN, HA, PCIII and CIV of the patients with a stage 2 liver fibrosis were 110 ng/ml, 110 ng/ml, 100 ng/ml and 70 ng/ml respectively, with sensibilities of diagnosing stage 2 liver fibrosis at 70%, 79%, 79% and 74% respectively. Their specificities in diagnosing stage 2 liver fibrosis were 68%, 72%, 64% and 73% respectively. The levels of LN, HA, PCIII and CIV in serum of these patients diagnosing cut-off value in stage 4 liver fibrosis (early cirrhosis) were 130 ng/ml, 140 ng/ml, 120 ng/ml and 70 ng/ml respectively. Their sensibility of diagnosing liver cirrhosis was 79%, 93%, 79% and 86% respectively. Their specificity of diagnosing liver cirrhosis was 66%, 82%, 72% and 61% respectively. As shown by the ROC curves in these patients, differentiating patients with cirrhosis or without cirrhosis, serum HA level was more valuable than LN, PCIII, CIV (the areas under the curves = 0.938 vs 0.775, 0.787, 0.791 ) When serum HA was higher than 190 ng/ml, the veracity of diagnosing liver cirrhosis was 93%. CONCLUSIONS: There is a certain quantitative relationship between the levels of LN, HA, PCIII and CIV in serum and the degrees of liver tissue fibrosis. The level of HA in serum is an important reference datum for early diagnosing liver cirrhosis. PMID- 16381638 TI - [Effects of metformin on fatty liver in insulin-resistant rats]. AB - OBJECTIVES: To investigate the effects of metformin on fatty livers in insulin resistant rats. METHODS: Thirty-one male Wistar rats were randomly divided into a normal (n=8) and an experimental group (n=23). The normal group rats were fed a standard diet, and those of the experimental group with a high-fat diet. After 8 weeks, 7 rats from the experimental group were sacrificed to verify whether the model was established successfully. Then the experimental group was randomly subdivided into two groups: one with metformin treatment (n=8) and one without (n=8). After 6 weeks, insulin sensitivity was measured with glucose infusion rate (GIR) by euglycermic hyperinsulinemia clamp technique. Aminotransferase, triglyceride (TG) and free fatty acids (FFA) were measured by biochemical methods, insulin by radioimmunoassay and tumor necrosis factor-alpha (TNF alpha) by ELISA. The steatosis changes and inflammation activity of all rat livers were scored histologically. RESULTS: Compared with the model group, the insulin resistance, liver index, visceral adiposity and weight loss of the metformin group were dramatically ameliorated. The steatosis and the inflammatory activity in the livers and the level of serum aminotransferase of the metformin group were also significantly decreased. Furthermore, metformin treatment lowered serum TG, liver lipid accumulation and the production of FFA and TNF alpha. CONCLUSION: Metformin can significantly improved insulin resistance and fatty liver development in rats fed a high-fat diet. It may become a new choice for fatty liver treatment in the future. PMID- 16381639 TI - [Clinical characteristics and ultrastructural features of livers in children with Wilson disease manifested mainly as hepatic injuries]. AB - OBJECTIVES: To study the feasibility and possibility to diagnose Wilson disease with electronmicroscopical examination of liver biopsies. METHODS: Clinical analysis, histological observation and ultrastructural examination were performed on 15 children with Wilson disease. RESULTS: All 15 subjects had symptoms of hepatic disorders, such as jaundice. Morphological signs of hepatocyte injury in three phase, namely steatosis, mitochondrion changes and cholestasis in bile canaliculi of the early phase, nucleus injury, dilation of endoplasmic reticulum, increase of lysosomes and appearance of residual bodies of the second phase, and massive autophagy and cirrhosis of the late phase were shown. A few inflammatory cells in the liver specimens were observed. Accumulation of copper in lysosomes and autophagosomes was found by energy-dispersion X-ray. CONCLUSION: The diagnostic signs for Wilson disease are autophagosomes in hepatocytes, cirrhosis accompanied with a few of inflammatory cells. A certain diagnosis of the disease depends on the finding of copper accumulation in hepatocytes. PMID- 16381640 TI - [Effects on intracellular Ca2+ and expression of L-type voltage-operated calcium channel protein in activated hepatic stellate cells stimulated by Chinese herbal compound 861]. PMID- 16381641 TI - [Inhibition of mouse hepatocyte apoptosis by anti-caspase-12 small interfering RNA]. AB - OBJECTIVES: To study the inhibition of primary mouse hepatocyte apoptosis by small interfering RNA (siRNAs) against caspase-12. METHODS: The Balb/c mouse primary hepatocytes were isolated in situ with two-step liver perfusion with 0.5 g/L collagenase type IV, and apoptosis were induced with 4 micromol/L thapsigargin (TG). The three kingds of siRNAs targeting different gene sites (130, 214, 521) were synthetized chemically. The single-stranded RNAs were annealed to produce double-stranded siRNAs, then the mouse primary hepatocytes were transfected by oligofectamine package. The inhibition of caspase-12 was analyzed with RT-PCR and Western-blot. The viable hepatocytes following the induction of apoptosis were evaluated with MTT. RESULTS: All the three kinds of siRNAs could obviously inhibit normal mouse hepatocyte caspase-12 mRNA. The siRNA (214) were more effective than the other two when the concentration was 100 nmol/L. The caspase-12 mRNA expression was inhibited by 52.08%, while that of siRNA (521) was 30.73% (t=4.30, P <0.05). However when the concentration was 200 nmol/L, the inhibitions were similar (88.07%, 86.22% and 89.41% respectively). siRNA (214) could downregulate the expression of apoptotic hepatocytes procaspase 12 by 51.43% ( t=4.30, P <0.01). Contrasted with apoptotic hepatocytes, the cell activity, which was analyzed with MTT, increased by 48.76% (t=2.23, P <0.01). CONCLUSION: siRNAs could effectively downregulate the expression of caspase-12 at mRNA and protein levels and prevent mouse primary hepatocytes from apoptosis. PMID- 16381642 TI - [Clinical and laboratory characteristics of anti-soluble liver antigen/liver pancreas (SLA/LP) autoantibody positive liver disease patients]. AB - OBJECTIVE: To analyze the clinical and laboratory features of anti-soluble liver antigen/liver-pancreas (SLA/LP) autoantibody positive patients with abnormal liver functions. METHODS: From July 1999 to August 2004, 4928 serum samples from patients with abnormal liver functions (ALT >40 U/L) were collected. A series of autoantibody examinations were carried out. Clinical manifestations and laboratory findings of 8 patients with anti-SLA/LP autoantibody positive were reviewed. RESULTS: Among the 5500 serum samples, 8 cases (6 females and 2 males) with positive anti-SLA/LP autoantibodies were found with complete clinical information. The age of the patients was (27-76) years old. The case histories were from 2 years to 10 years. Of the 8 patients, 6 cases had liver cirrhosis and HBsAg-negative and anti-HCV-negative, active, 1 case had liver cirrhosis with HBsAg-positive, but HBVDNA negative; 1 case had liver cirrhosis and anti-HCV positive, but HCV RNA negative. The 8 cases were all ANA positive with titers of 31:320. Four cases were AMA positive and 2 among these 4 cases were M2 positive. The most frequent symptoms were fatigue, anorexia, nausea, jaundice, abdominal distention and edema of lower limbs. All patients had high hypergammaglobulinemia. CONCLUSION: Anti-SLA/LP autoantibody was at a low detection rate in the study with females in preponderance, Clinical and laboratory characteristics of the 8 cases were consistent with those of the autoimmune hepatitis (AIH). Testing for anti-SLA autoantibodies helps in the diagnosis of AIH in many patients who may otherwise be misdiagnosed. PMID- 16381643 TI - [A pathological study on liver tissues of patients with HIV infection]. AB - OBJECTIVE: To study the pathological changes of the liver tissues of patients with HIV infection. METHODS: 14 biopsy and 12 autopsy liver tissues were examined histologically. HIV-1 related antigen of outer membrane protein gp120 and capsid protein p24 were examined with their corresponding monoclonal antibodies by immunohistochemistry. RESULTS: In the biopsy group, cytomegalic virus (CMV) infection was found in one (1/14) case, outer membrane protein gp120 and/or capsid protein p24 antigen were detected in Kupffer cells and in some of the lymphocytes in 11 cases. All the hepatocytes were negative for outer membrane protein gp120 and capsid protein p24 antigens. In the autopsy group, there were 5 (5/12) cases of liver tissues with CMV infection and 5 cases each with mycobacterium and Toxoplasma gondii infection. Capsid protein p24 was detected in liver tissues in 3 cases. CONCLUSION: There is HIV infection in liver tissue of patients with HIV. The rate of opportunistic infections in liver biopsy samples was lower than that in the autopsy liver tissues of patients with HIV. PMID- 16381644 TI - [Effect of a Chinese herbal compound and lamivudine combination therapy on patients with chronic hepatitis B and its influence on YMDD motif mutation]. PMID- 16381645 TI - [Treatment of CCl4 induced chronic liver injury with bone marrow mesenchymal stem cells overexpressing hepatocyte growth factor]. PMID- 16381646 TI - [Expression of HBsAg and HBcAg in the ovaries of patients with chronic hepatitis B]. PMID- 16381647 TI - [The effect of intestinal endotoxemia on the balance of Th1/Th2 in patients with hepatitis B]. PMID- 16381648 TI - [Treatment of severe liver damage with autologous bone marrow stem cell transplantation in human patients]. PMID- 16381649 TI - [The expression of B7 and CD28 in peripheral blood mononuclear cells of chronic severe hepatitis B patients and its clinical significance]. PMID- 16381650 TI - [Clinical application of endoscopic esophageal variceal ligation combined with partial splenic embolization on patients with portal hypertension]. PMID- 16381651 TI - [Two cases report and review of hepatic veno-occlusive disease]. PMID- 16381652 TI - [A report from the Second China National Annual Meeting of Artificial Liver and Blood Purification]. PMID- 16381653 TI - [The regulatory mechanism of liver oval cell differentiation]. PMID- 16381654 TI - [HIV/HBV coinfection]. PMID- 16381655 TI - [Subacute severe hepatitis as the initial presentation of autoimmune hepatitis: report of 2 cases]. PMID- 16381656 TI - [Progress of the study on the relationship between MxA and interferon therapy of viral hepatitis]. PMID- 16381657 TI - Structural diversity of cytosolic free oligosaccharides in the human hepatoma cell line, HepG2. AB - It is thought that free oligosaccharides in the cytosol are an outcome of quality control of glycoproteins by endoplasmic reticulum-associated degradation (ERAD). Although considerable amounts of free oligosaccharides accumulate in the cytosol, where they presumably have some function, detailed analyses of their structures have not yet been carried out. We isolated 21 oligosaccharides from the cytosolic fraction of HepG2 cells and analyzed their structures by the two-dimensional high performance liquid chromatography (HPLC) sugar-mapping method. Sixteen novel oligosaccharides were identified in the cytosol in this study. All had a single N acetylglucosamine at their reducing-end cores and could be expressed as (Man)n (GlcNAc)1. No free oligosaccharide with N,N'-diacetylchitobiose was detected in the cytosolic fraction of HepG2 cells. This suggested that endo-beta-N acetylglucosaminidase was a key enzyme in the production of cytosolic free oligosaccharides. The 21 oligosaccharides were classified into three series- series 1: oligosaccharides processed from Manalpha1-2Manalpha1-6 (Manalpha1 2Manalpha1-3)Manalpha1-6(Manalpha1-2Manalpha1-2Manalpha1-3) Manbeta1-4GlcNAc (M9A') and Manalpha1-2Manalpha1-6(Manalpha1-3) Manalpha1-6(Manalpha1-2Manalpha1 2Manalpha1-3)Manbeta1-4GlcNAc (M8A') by digestion with cytosolic alpha mannosidase; series 2: oligosaccharides processed with Golgi alpha-mannosidases in addition to endoplasmic reticulum (ER) and cytosolic alpha-mannosidases; and series 3: glucosylated oligosaccharides produced from Glc1Man9GlcNAc1 by hydrolysis with cytosolic alpha-mannosidase. The presence of the series "2" oligosaccharides suggests that some of the misfolded glycoproteins had been processed in pre-cis-Golgi vesicles and/or the Golgi apparatus. When the cells were treated with swainsonine to inhibit cytosolic alpha-mannosidase, the amounts of M9A' and M8A' increased remarkably, suggesting that these oligosaccharides were translocated into the cytosol. Four oligosaccharides of series "2" also increased. In contrast, there were obvious reductions in Manalpha1-6(Manalpha1 2Manalpha1-2Manalpha1-3)Manbeta1-4GlcNAc (M5B'), the end product from M9A' by digestion with cytosolic alpha-mannosidase, and Manalpha1-6(Manalpha1- 2Manalpha1 3)Manbeta1-4GlcNAc, derived from series "2" oligosaccharides by digestion with cytosolic alpha-mannosidase. Our data suggest that (1) some of the cytosolic oligosaccharides had been processed with Golgi alpha-mannosidases, (2) the major oligosaccharides translocated from the ER were M9A' and M8A', and (3) M5B' and Glc1M5B' were maintained at relatively high concentrations in the cytosol. PMID- 16381658 TI - A highly efficient transformation protocol for Micro-Tom, a model cultivar for tomato functional genomics. AB - We report a highly efficient protocol for the Agrobacterium-mediated genetic transformation of a miniature dwarf tomato (Lycopersicon esculentum), Micro-Tom, a model cultivar for tomato functional genomics. Cotyledon explants of tomato inoculated with Agrobacterium tumefaciens (Rhizobium radiobacter) C58C1Rif(R) harboring the binary vector pIG121Hm generated a mass of chimeric non-transgenic and transgenic adventitious buds. Repeated shoot elongation from the mass of adventitious buds on selection media resulted in the production of multiple transgenic plants that originated from independent transformation events. The transformation efficiency exceeded 40% of the explants. This protocol could become a powerful tool for functional genomics in tomato. PMID- 16381659 TI - Interspecific pairs of class II S haplotypes having different recognition specificities between Brassica oleracea and Brassica rapa. AB - There are several pairs of similar class I S haplotypes between Brassica oleracea and Brassica rapa. The similar S halotypes in these interspecific pairs have been reported to have the same recognition specificities. In the present study, three interspecific pairs showing a high sequence similarity were found in class II S haplotypes, i.e. between BoS-2b (B. oleracea S-2b) and BrS-44 (B. rapa S-44), between BoS-5 and BrS-40, and between BoS-15 and BrS-60. By pollination tests using interspecific hybrids between B. oleracea and B. rapa, BoS-5 and BoS-2b were revealed to have slightly and completely different recognition specificities from those of BrS-40 and BrS-44, respectively. The recognition reaction between SP11 and SRK of BoS-15 was suggested to be incomplete. The regions of class II SP11 and SRK important for self-recognition specificity and the diversification of class II S haplotypes are discussed herein. PMID- 16381660 TI - Endocrine disrupting chemical atrazine causes degranulation through Gq/11 protein coupled neurosteroid receptor in mast cells. AB - We studied the effects of representative endocrine-disrupting chemicals on beta hexosaminidase release from mast cells and their putative neurosteroid receptor involvement. Some endocrine-disrupting chemicals, such as amitrol, benzophenon, bisphenol A, pentachlorophenol, and tetrabromophenol A did not cause hexosaminidase release from RBL-2H3 cells, but they blocked the release by dehydroepiandrosterone sulfate, a representative neurosteroid agonist. On the contrary, atrazine, which is a widely used herbicide, caused a rapid and concentration-dependent degranulation in the range between 10 nM and 1 microM in RBL-2H3 and peritoneal mast cells. Atrazine-induced degranulation was also evaluated by Alexa 488-annexin V binding to the phosphatidylserine, which is externalized during degranulation, and these actions were blocked by BSA conjugated (membrane-impermeable) progesterone (PROG-BSA). The atrazine-induced beta-hexosaminidase release was characterized by various inhibitors including antisense-oligodeoxynucleotide for Galpha(q/11), pertussis toxin, phospholipase C inhibitor U-73122, inositol 1,4,5-triphosphate receptor inhibitor xestospongin C and Ca(2+) channel blocker lanthanum chloride. These analyses revealed that the degranulation is mediated by putative metabotropic neurosteroid receptor, G(q/11), phospholipase C and Ca(2+) mobilization from intracellular stores. Having documented progesterone receptor-modulation of atrazine-induced mast cell degranulation in vitro, this response was evaluated in mice. Atrazine caused pain responses when injected in the foot pads of mice, and they were antagonized by local administration of PROG-BSA or diphenhydramine. Atrazine also caused PROG BSA-reversible plasma extravasation. All these findings strongly suggest that herbicide atrazine exerts inflammatory activity through activation of putative G(q/11)-coupled neurosteroid receptor and phospholipase C. PMID- 16381661 TI - Characterizing the ovotoxicity of cyclophosphamide metabolites on cultured mouse ovaries. AB - Cyclophosphamide (CPA) is reported to target dormant primordial ovarian follicles in rodents and humans. However, mechanistic studies are complicated due to the complex ovarian structure. We present here the characterization of the sensitivity of ovaries to CPA metabolites and the timing of morphological alterations induced by phosphoramide mustard (PM) in an in vitro system. Intact mouse ovaries (postnatal-day-4) were cultured in vitro and exposed to multiple breakdown products of CPA on day 0 (d0). Tissues were cultured up to d8, and then follicle counts and immunohistochemistry were performed. 4-Hydroperoxy-CPA (4 HC), a precursor of an activated form of CPA, and PM depleted primordial and primary follicles (> or =1 microM and > or =3 microM, respectively, p < 0.05); acrolein had effects on follicle numbers only under continuous exposure (> =30 microM); carboxycyclophosphamide and 4-ketocyclophosphamide reduced primordial and small primary follicles only at high concentrations (100 microM). PM-induced follicle loss became significant (p < 0.05) by d1 or d2 following exposures to 10 microM or 3 microM PM, respectively, as determined by the numbers of pyknotic or TUNEL-positive follicles. Cellular targets were oocytes in the smallest follicles, but granulosa cells in large primary follicles. TUNEL staining was observed in both cell types, but caspase-3, a marker of apoptosis, was absent from primordial follicles. In addition, a pan-caspase inhibitor could not prevent follicle losses when administered prior to PM. Thus, brief exposures to 4-HC or PM are sufficient to induce permanent follicle loss in ovaries, and PM is likely the ultimate ovotoxicant. Furthermore, the cell death pathway is likely caspase independent. PMID- 16381662 TI - A combined approach to drug metabolism and toxicity assessment. AB - The challenge of predicting the metabolism or toxicity of a drug in humans has been approached using in vivo animal models, in vitro systems, high throughput genomics and proteomics methods, and, more recently, computational approaches. Understanding the complexity of biological systems requires a broader perspective rather than focusing on just one method in isolation for prediction. Multiple methods may therefore be necessary and combined for a more accurate prediction. In the field of drug metabolism and toxicology, we have seen the growth, in recent years, of computational quantitative structure-activity relationships (QSARs), as well as empirical data from microarrays. In the current study we have further developed a novel computational approach, MetaDrug, that 1) predicts metabolites for molecules based on their chemical structure, 2) predicts the activity of the original compound and its metabolites with various absorption, distribution, metabolism, excretion, and toxicity models, 3) incorporates the predictions with human cell signaling and metabolic pathways and networks, and 4) integrates networks and metabolites, with relevant toxicogenomic or other high throughput data. We have demonstrated the utility of such an approach using recently published data from in vitro metabolism and microarray studies for aprepitant, 2(S)-((3,5-bis(trifluoromethyl)benzyl)-oxy)-3(S)phenyl-4-((3-oxo 1,2,4-triazol-5-yl)methyl)morpholine (L-742694), trovofloxacin, 4 hydroxytamoxifen, and artemisinin and other artemisinin analogs to show the predicted interactions with cytochromes P450, pregnane X receptor, and P glycoprotein, and the metabolites and the networks of genes that are affected. As a comparison, we used a second computational approach, MetaCore, to generate statistically significant gene networks with the available expression data. These case studies demonstrate the combination of QSARs and systems biology methods. PMID- 16381663 TI - Purification and characterization of akr1b10 from human liver: role in carbonyl reduction of xenobiotics. AB - Members of the aldo-keto reductase (AKR) superfamily have a broad substrate specificity in catalyzing the reduction of carbonyl group-containing xenobiotics. In the present investigation, a member of the aldose reductase subfamily, AKR1B10, was purified from human liver cytosol. This is the first time AKR1B10 has been purified in its native form. AKR1B10 showed a molecular mass of 35 kDa upon gel filtration and SDS-polyacrylamide gel electrophoresis. Kinetic parameters for the NADPH-dependent reduction of the antiemetic 5-HT3 receptor antagonist dolasetron, the antitumor drugs daunorubicin and oracin, and the carcinogen 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) to the corresponding alcohols have been determined by HPLC. Km values ranged between 0.06 mM for dolasetron and 1.1 mM for daunorubicin. Enzymatic efficiencies calculated as kcat/Km were more than 100 mM-1 min-1 for dolasetron and 1.3, 0.43, and 0.47 mM-1 min-1 for daunorubicin, oracin, and NNK, respectively. Thus, AKR1B10 is one of the most significant reductases in the activation of dolasetron. In addition to its reducing activity, AKR1B10 catalyzed the NADP+ dependent oxidation of the secondary alcohol (S)-1-indanol to 1-indanone with high enzymatic efficiency (kcat/Km=112 mM-1 min-1). The gene encoding AKR1B10 was cloned from a human liver cDNA library and the recombinant enzyme was purified. Kinetic studies revealed lower activity of the recombinant compared with the native form. Immunoblot studies indicated large interindividual variations in the expression of AKR1B10 in human liver. Since carbonyl reduction of xenobiotics often leads to their inactivation, AKR1B10 may play a role in the occurrence of chemoresistance of tumors toward carbonyl group-bearing cytostatic drugs. PMID- 16381664 TI - Modulation of the P-glycoprotein-mediated intestinal secretion of ivermectin: in vitro and in vivo assessments. AB - The everted gut sac method was used to assess the role of the P-glycoprotein (P gp) on the intestinal secretion of ivermectin (IVM), an antiparasitic widely used in human and veterinary medicine. The work included the evaluation of two different P-gp modulators [itraconazole (ITZ) and valspodar (PSC833)] used at equimolar doses in the rat. Furthermore, the influence of both P-gp modulator agents on the disposition kinetics of IVM in plasma, liver, and gastrointestinal tissues was characterized. For the in vitro experiments, ileal sacs were incubated with IVM (3 microM) in the presence or absence of either ITZ (10 microM) or PSC833 (10 microM). In the in vivo experiments, male Wistar rats were randomly allocated to three groups (n=18) and subcutaneously treated with IVM (200 microg/kg-1), alone and coadministered with ITZ (5 mg, two doses) or PSC833 (8.6 mg, two doses). Animals were sacrificed between 6 and 96 h. Blood, liver, and gastrointestinal samples were collected. IVM concentrations were determined by high performance liquid chromatography. The rate of IVM accumulation in the intestinal wall of everted sacs was significantly higher after its incubation with ITZ (0.115 nmol/g/min) and PSC833 (0.238 nmol/g/min) than that obtained after the incubation without the P-gp modulators (0.016 nmol/g/min). In agreement with the in vitro experiment, the presence of ITZ and PSC833 induced an enhancement in the concentrations of IVM in plasma and gastrointestinal tissues. The results obtained in the current work, both under in vivo and in vitro conditions, confirm the relevance of P-gp-mediated transport to the intestinal secretion of IVM. PMID- 16381665 TI - Pharmacokinetics and metabolism of a selective androgen receptor modulator in rats: implication of molecular properties and intensive metabolic profile to investigate ideal pharmacokinetic characteristics of a propanamide in preclinical study. AB - S-1 [3-(4-fluorophenoxy)-2-hydroxy-2-methyl-N-[4-nitro-3-(trifluoromethyl)phenyl] propanamide] is one member of a series of potent selective androgen receptor modulators (SARMs) that are being explored and developed for androgen-dependent diseases. Recent studies showed that S-1 holds great promise as a novel therapeutic agent for benign hyperplasia [W. Gao, J. D. Kearbey, V. A. Nair, K. Chung, A. F. Parlow, D. D. Miller, and J. T. Dalton (2004) Endocrinology 145:5420 5428]. We examined the pharmacokinetics and metabolism of S-1 in rats as a component of our preclinical development of this compound and continued interest in structure-activation relationships for SARM action. Forty male Sprague-Dawley rats were randomly assigned to treatment groups and received either an i.v. or a p.o. dose of S-1 at a dose level of 0.1, 1, 10, or 30 mg/kg. S-1 demonstrated a low clearance (range, 3.6-5.2 ml/min/kg), a moderate volume of distribution (range, 1460-1560 ml/kg), and a terminal half-life ranging from 3.6 to 5.2 h after i.v. doses. The oral bioavailability of S-1 ranged from 55% to 60%. Forty phase I and phase II metabolites of S-1 were identified in the urine and feces of male Sprague-Dawley rats dosed at 50 mg/kg via the i.v. route. The two major urinary metabolites of S-1 were a carboxylic acid and a sulfate-conjugate of 4 nitro-3-trifluoromethylphenylamine. Phase I metabolites arising from A-ring nitro reduction to an aromatic amine and B-ring hydroxylation were also identified in the urinary and fecal samples of rats. Furthermore, a variety of phase II metabolites through sulfation, glucuronidation, and methylation were also found. These studies demonstrate that S-1 is rapidly absorbed, slowly cleared, moderately distributed, and extensively metabolized in rats. PMID- 16381666 TI - Metabolism and excretion of erlotinib, a small molecule inhibitor of epidermal growth factor receptor tyrosine kinase, in healthy male volunteers. AB - Metabolism and excretion of erlotinib, an orally active inhibitor of epidermal growth factor receptor tyrosine kinase, were studied in healthy male volunteers after a single oral dose of [14C]erlotinib hydrochloride (100-mg free base equivalent, approximately 91 microCi/subject). The mass balance was achieved with approximately 91% of the administered dose recovered in urine and feces. The majority of the total administered radioactivity was excreted in feces (83+/ 6.8%), and only a low percentage of the dose was recovered in urine (8.1+/-2.8%). Only less than 2% of what was recovered in humans was unchanged erlotinib, which demonstrates that erlotinib is eliminated predominantly by metabolism. In plasma, unchanged erlotinib represented the major circulating component, with the pharmacologically active metabolite M14 accounting for approximately 5% of the total circulating radioactivity. Three major biotransformation pathways of erlotinib are O-demethylation of the side chains followed by oxidation to a carboxylic acid, M11 (29.4% of dose); oxidation of the acetylene moiety to a carboxylic acid, M6 (21.0%); and hydroxylation of the aromatic ring to M16 (9.6%). In addition, O-demethylation of M6 to M2, O-demethylation of the side chains to M13 and M14, and conjugation of the oxidative metabolites with glucuronic acid (M3, M8, and M18) and sulfuric acid (M9) play a minor role in the metabolism of erlotinib. The identified metabolites accounted for >90% of the total radioactivity recovered in urine and feces. The metabolites observed in humans were similar to those found in the toxicity species, rats and dogs. PMID- 16381667 TI - Glucuronidation as a major metabolic clearance pathway of 14c-labeled muraglitazar in humans: metabolic profiles in subjects with or without bile collection. AB - The metabolism and disposition of 14C-labeled muraglitazar (Pargluva), a novel dual alpha/gamma peroxisome proliferator-activated receptor activator, was investigated in eight healthy male subjects with and without bile collection (groups 1 and 2) after a single 20-mg oral dose. Bile samples were collected for 3 to 8 h after dosing from group 2 subjects in addition to the urine and feces collection. In plasma, the parent compound was the major component, and circulating metabolites, including several glucuronide conjugates, were minor components at all time points. The exposure to parent drug (Cmax and area under the plasma concentration versus time curve) in subjects with bile collection was generally lower than that in subjects without bile collection. The major portion of the radioactive dose was recovered in feces (91% for group 1 and 51% for group 2). In addition, 40% of the dose was recovered in the bile from group 2 subjects. In this 3- to 8-h bile, the glucuronide of muraglitazar (M13, 15% of dose) and the glucuronides of its oxidative metabolites (M17a,b,c, M18a,b,c, and M20, together, 16% of dose) accounted for approximately 80% of the biliary radioactivity; muraglitazar and its O-demethylated metabolite (M15) each accounted for approximately 4% of the dose. In contrast, fecal samples only contained muraglitazar and its oxidative metabolites, suggesting hydrolysis of biliary glucuronides in the intestine before fecal excretion. Thus, the subjects with and without bile collection showed different metabolic profiles of muraglitazar after oral administration, and glucuronidation was not observed as a major pathway of metabolic clearance from subjects with the conventional urine and fecal collection, but was found as a major elimination pathway from subjects with bile collection. PMID- 16381668 TI - Selectivity of substrate (trifluoperazine) and inhibitor (amitriptyline, androsterone, canrenoic acid, hecogenin, phenylbutazone, quinidine, quinine, and sulfinpyrazone) "probes" for human udp-glucuronosyltransferases. AB - Relatively few selective substrate and inhibitor probes have been identified for human UDP-glucuronosyltransferases (UGTs). This work investigated the selectivity of trifluoperazine (TFP), as a substrate, and amitriptyline, androsterone, canrenoic acid, hecogenin, phenylbutazone, quinidine, quinine, and sulfinpyrazone, as inhibitors, for human UGTs. Selectivity was assessed using UGTs 1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B7, and 2B15 expressed in HEK293 cells. TFP was confirmed as a highly selective substrate for UGT1A4. However, TFP bound extensively to both HEK293 lysate and human liver microsomes in a concentration-dependent manner (fuinc 0.20-0.59). When corrected for nonspecific binding, Km values for TFP glucuronidation were similar for both UGT1A4 (4.1 microM) and human liver microsomes (6.1+/-1.2 microM) as the enzyme sources. Of the compounds screened as inhibitors, hecogenin, alone, was selective; significant inhibition was observed only for UGT1A4 (IC50 1.5 microM). Using phenylbutazone and quinine as "models," inhibition kinetics were variously described by competitive and noncompetitive mechanisms. Inhibition of UGT2B7 by quinidine was also investigated further, because the effects of this compound on morphine pharmacokinetics (a known UGT2B7 substrate) have been ascribed to inhibition of P-glycoprotein. Quinidine inhibited human liver microsomal and recombinant UGT2B7, with respective Ki values of 335+/-128 microM and 186 microM. In conclusion, TFP and hecogenin represent selective substrate and inhibitor probes for UGT1A4, although the extensive nonselective binding of the former should be taken into account in kinetic studies. Amitriptyline, androsterone, canrenoic acid, hecogenin, phenylbutazone, quinidine, quinine, and sulfinpyrazone are nonselective UGT inhibitors. PMID- 16381669 TI - Novel metabolites of buprenorphine detected in human liver microsomes and human urine. AB - The in vitro metabolism of buprenorphine was investigated to explore new metabolic pathways and identify the cytochromes P450 (P450s) responsible for the formation of these metabolites. The resulting metabolites were identified by liquid chromatography-electrospray ionization-tandem mass spectrometry. In addition to norbuprenorphine, two hydroxylated buprenorphine (M1 and M2) and three hydroxylated norbuprenorphine (M3, M4, and M5) metabolites were produced by human liver microsomes (HLMs), with hydroxylation occurring at the tert-butyl group (M1 and M3) and at unspecified site(s) on the ring moieties (M2, M4, and M5). Time course and other data suggest that buprenorphine is N-dealkylated to form norbuprenorphine, followed by hydroxylation to form M3; buprenorphine is hydroxylated to form M1 and M2, followed by N-dealkylation to form M3 and M4 or M5. The involvement of selected P450s was investigated using cDNA-expressed P450s coupled with scaling models, chemical inhibition, monoclonal antibody (MAb) analysis, and correlation studies. The major enzymes involved in buprenorphine elimination and norbuprenorphine and M1 formation were P450s 3A4, 3A5, 3A7, and 2C8, whereas 3A4, 3A5, and 3A7 produced M3 and M5. Based on MAb analysis and chemical inhibition, the contribution of 2C8 was higher in HLMs with higher 2C8 activity, whereas 3A4/5 played a more important role in HLMs with higher 3A4/5 activity. Examination of human urine from subjects taking buprenorphine showed the presence of M1 and M3; most of M1 was conjugated, whereas 60 to 70% of M3 was unconjugated. PMID- 16381670 TI - Benzo[A]pyrene-induced oral carcinogenesis and chemoprevention: studies in bioengineered human tissue. AB - Oral cancer, originating from smoking-induced lesions of the basal cells in the complex stratified oral epithelium, is difficult to treat. Early detection of premalignant lesions, e.g., leukoplakia, has suggested the possibility of chemopreventive measures, such as topical application of antimutagenic/antiproliferative dietary or pharmaceutical agents. As an extension of a study in human oral epithelial cell monolayers, we determined the carcinogen, i.e., benzo[a]pyrene (BaP), transport, bioactivation, and DNA binding in a bioengineered human gingival epithelial tissue construct and the chemopreventive effects of dietary polyphenols. Short-term experiments showed that both types of compounds can traverse this tissue as well as be effectively taken up by the tissue. The model cigarette smoke carcinogen BaP very slowly, but to a great extent, accumulated in the tissue with maximal uptake at 24 h. Such exposure clearly resulted in DNA binding of BaP by the tissue. This DNA binding was associated with BaP-induced CYP1B1 as well as CYP1A1 expression, as evidenced by mRNA measurements. Cotreatment of the oral tissue with dietary polyphenols, including resveratrol and quercetin, and BaP, resulted in significant inhibition of the BaP-DNA binding. Using fluorescence microscopy as well as simultaneous autoradiography, we also demonstrated that quercetin indeed penetrates the entire stratified tissue layer, but that quercetin was also oxidized within the cells. Thus, this bioengineered oral tissue construct opens up improved ways of understanding and preventing/treating smoking-induced oral cancer. PMID- 16381671 TI - Tissue distribution and ontogeny of organic cation transporters in mice. AB - Organic cation transporters (Octs) play an important role in transporting cationic xeno- and endobiotics across biological membranes. Little is known about Octs in mice; therefore, the tissue distribution and developmental changes in the mRNA expression of Octs in mice were quantified. Oct1, Oct2, Oct3, Octn1, Octn2, and Octn3 mRNA expression was quantified in 14 tissues from male and female mice using the branched DNA signal amplification assay. Oct1 mRNA expression was highest in kidney, followed by liver. Oct2 mRNA was almost exclusively expressed in kidney, with male mice having twice that in female mice. The higher expression of Oct2 in male mice is due to testosterone. Oct3 mRNA was most highly expressed in placenta, ovary, and uterus, but was expressed at low levels in most tissues. Octn1 and Octn2 mRNA expression was similar, with the highest levels in kidney followed by small intestine. Octn3 mRNA was almost exclusively expressed in testes. The developmental expression of Oct1, Oct2, Octn1, and Octn2 mRNA in kidneys as well as Oct1 in liver was determined in young mice. Ontogenic expression data indicate that each of the Octs approached adult expression levels by about 3 weeks of age. The gender difference in Oct mRNA expression did not become apparent until day 30 after birth. The differences in tissue distribution of the Octs may play an important role in drug disposition to various tissues. Furthermore, low expression of the Octs in young animals may affect the pharmacokinetic behavior of drugs compared with that in adults. PMID- 16381672 TI - Sulfation of raloxifene and 4-hydroxytamoxifen by human cytosolic sulfotransferases. AB - Raloxifene and 4-hydroxytamoxifen (4-OHT) are important estrogen-related drugs used in the treatment of osteoporosis and breast cancer. Sulfation is involved in the metabolism and inactivation of both compounds in human tissues, although the sulfotransferase (SULT) isoforms involved in their conjugation have not been well described. The ability of seven expressed SULT isoforms to sulfate raloxifene and 4-OHT was investigated. Raloxifene was conjugated by all seven SULT isoforms tested, whereas 4-OHT was conjugated only by SULTs 1A1, 1E1, and 2A1. Characterization of raloxifene and 4-OHT sulfation demonstrates that sulfation can occur at therapeutic concentrations. SULT1E1 displayed the lowest Km (0.2 microM) for 4-OHT sulfation and SULT2A1 the lowest (0.3 microM) for raloxifene sulfation. SULT1E1 was the only isoform exhibiting detectable levels of raloxifene disulfation activity. Modeling of the interactions of raloxifene in the active site of SULT1E1 indicates that both hydroxyl groups of raloxifene can be readily positioned in proximity to the sulfonyl group of 3'-phosphoadenosine 5'-phosphosulfate and the catalytically important His107 residue. Both raloxifene and 4-OHT sulfation activities were detectable in all human liver cytosols tested. 4-OHT sulfation was detected in cytosol prepared from endometrial biopsies of normal women obtained during the proliferative and secretory phases of the same menstrual cycle. In contrast, raloxifene sulfation was detectable only in secretory phase cytosols in association with SULT1E1 activity. In summary, several human SULT isoforms are capable of sulfating raloxifene and 4 OHT. Tissue-specific expression of the individual SULT isoforms may have important roles in the regulation of the activity of these compounds. PMID- 16381673 TI - The role of pregnane X receptor in 2-acetylaminofluorene-mediated induction of drug transport and -metabolizing enzymes in mice. AB - Activation of the pregnane X receptor (PXR) mediates the induction of several drug transporters and -metabolizing enzymes. In vitro studies have reported that several of these genes are induced after exposure to the hepatocarcinogen, 2 acetylaminofluorene (2-AAF). Thus, we hypothesized that PXR may play a role in the in vivo induction of gene expression by 2-AAF. We examined the expression of the drug-metabolizing enzymes CYP1A2 and CYP3A11 and the drug transporters breast cancer resistance protein (BCRP), MRP2, and OATP2. Wild-type (PXR+/+) and PXR null (PXR-/-) C57BL/6 mice were injected daily for 7 days with 150 or 300 mg/kg 2 AAF suspended in corn oil (i.p.), whereas the control group received corn oil vehicle. Levels of mRNA isolated from liver were measured by reverse transcription-polymerase chain reaction and normalized to beta-actin. Treatment of PXR+/+ mice resulted in a dose-dependent 2- to 4-fold induction (p<0.001) of MRP2, OATP2, BCRP, CYP3A11, and CYP1A2, but no induction was observed in PXR-/- mice. Induction of PXR mRNA was observed in the 2-AAF-treated PXR+/+ mice. Furthermore, a dose-dependent increase in CYP3A4 promoter construct activity was observed in HepG2 cells cotransfected with human or rat PXR, indicating that 2 AAF does indeed activate PXR. These results suggest that PXR is responsible for 2 AAF-mediated induction of drug efflux transporters and biotransformation enzymes in the liver. Moreover, novel findings demonstrate that PXR plays a role in regulation of the drug efflux transporter, BCRP, in mice. PMID- 16381675 TI - Nanodosimetry, from radiation physics to radiation biology. AB - In view of the fact that early damage to genes and cells by ionising radiation starts with the early damage to segments of the DNA, it is a great challenge to radiation research to describe the general behaviour of ionising radiation in nanometric target volumes (nanodosimetry). After summarising basic aspects of nanodosimetry, an overview is given about its present state. As far as experimental procedures are concerned, main emphasis is laid on single-ion counting and single-electron counting methods, which use millimetric target volumes filled with a low-pressure gas to simulate nanometric target volumes at unit density. Afterwards, physical principles are discussed, which can be used to convert experimental ionisation cluster-size distributions into those caused by ionising radiation in liquid water. In the final section, possibilities are analysed of how to relate parameters derived from the probability of cluster-size formation in liquid water to parameters derived from radiobiological experiments. PMID- 16381676 TI - 21st century challenges in radiation protection and shielding: draft 2005 recommendations of ICRP. AB - The International Commission on Radiological Protection regularly examines the status of its recommendations and reviews the increasing knowledge of the effects of exposure to ionising radiation in order to decide whether new recommendations are needed. The Commission strives to make its system more coherent and comprehensible, while recognising the need for stability in international and national regulations, many of which have, only fairly recently, implemented the 1990 Recommendations. However, new scientific data have been produced since 1990 and there have been societal developments in that more openness or transparency is expected in developing new recommendations and, in addition, there has been a move from the utilitarian approach of 'the greatest good for the greatest number', to one with more concern for the 'individual', all of which have inevitably led to some changes in the formulation of the recommendations. This paper outlines the proposals for the 2005 Recommendations. PMID- 16381677 TI - Radiation protection standards: their evolution from science to philosophy. AB - The concept of applying constraints on individual sources to a small fraction of the public dose limit has been deemed inappropriate when shielding the medical X ray sources. This represents a broad-based consensus of medical physics and radiological societies in the United States, and the report series on the shielding design for medical X-ray sources (including dental, X-ray imaging and therapeutic X ray) from the National Council on Radiation Protection and Measurements (NCRP) utilises 1 mSv y(-1) as a source control limit. In the present study, the rationale for such a conclusion is discussed, and a somewhat critical look at the current model of radiation protection of the public is made. PMID- 16381678 TI - The Spallation Neutron Source (SNS) project: a fertile ground for radiation protection and shielding challenges. AB - The Spallation Neutron Source facility presently under construction in the USA consists of a front end, a linac, an accumulator ring, a target station and a neutron instrument hall, producing pulsed neutron beams driven by a proton beam of 1 GeV energy and 1.4 MW power with a repetition rate of 60 Hz. The layout of the facility and the radiation protection and shielding concept of the facility is laid out in numerous examples in a walk from the proton beam generation to the neutron utilisation. PMID- 16381679 TI - High Intensity Proton Accelerator Project in Japan (J-PARC). AB - The High Intensity Proton Accelerator Project, named as J-PARC, was started on 1 April 2001 at Tokai-site of JAERI. The accelerator complex of J-PARC consists of three accelerators: 400 MeV Linac, 3 GeV rapid cycle synchrotron and 50 GeV synchrotron; and four major experimental facilities: Material and Life Science Facility, Nuclear and Particle Physics Facility, Nuclear Transmutation Experiment Facility and Neutrino Facility. The outline of the J-PARC is presented with the current status of construction. PMID- 16381680 TI - Implications of the space radiation environment for human exploration in deep space. AB - Human exploration of the solar system beyond Earth's orbit will entail many risks for the crew on these deep space missions. One of the most significant health risks is exposure to the harsh space radiation environment beyond the protection provided by the Earth's intrinsic magnetic field. Crew on exploration missions will be exposed to a complex mixture of very energetic particles. Chronic exposures to the ever-present background galactic cosmic ray (GCR) spectrum consisting of all naturally occurring chemical elements are combined with sporadic, possibly acute exposures to large fluxes of solar energetic particles, mainly protons and alpha particles. The background GCR environment is mainly a matter of concern for stochastic effects, such as the induction of cancer with subsequent mortality in many cases, and late deterministic effects, such as cataracts and possible damage to the central nervous system. Unfortunately, the actual risks of cancer induction and mortality owing to the very important high energy heavy ion component of the GCR spectrum are essentially unknown. The sporadic occurrence of extremely large solar energetic particle events (SPE), usually associated with intense solar activity, is also a major concern for the possible manifestation of acute effects from the accompanying high doses of such radiations, especially acute radiation syndrome effects such as nausea, emesis, haemorrhaging or, possibly, even death. In this presentation, an overview of the space radiation environment, estimates of the associated body organ doses and equivalent doses and the potential biological effects on crew in deep space are presented. Possible methods of mitigating these radiations, thereby reducing the associated risks to crew are also described. PMID- 16381681 TI - New trends in medical imaging. AB - In the past decades new imaging techniques and developments in existing systems have been introduced. Novel ideas with strong potential in areas such as radiography, nuclear magnetic resonance, ultrasound and nuclear medicine have recently emerged or are still under development or evaluation. Positron emission tomography (PET) is finally garnering enormous interest for high-sensitivity and high-specificity molecular imaging. The combination of PET and computed tomography is proving to be very useful in clinical oncology. Functional magnetic resonance imaging and magnetic resonance spectroscopy are also emerging as strong partners in terms of functional information. Recent improvements in ultrasound imaging, such as a 3-D, tissue harmonics, high-frequency, real time, extended field of view and new contrast agents, show the potential of this methodology. Film/screen mammography represents the ultimate in radiography capacities, but new digital systems are beginning to offer even better response. PMID- 16381682 TI - Properties of the Feynman-alpha method applied to accelerator-driven subcritical systems. AB - A Monte Carlo study of the Feynman-method with a simple code simulating the multiplication chain, confined to pertinent time-dependent phenomena has been done. The significance of its key parameters (detector efficiency and dead time, k-source and spallation neutrons multiplicities, required number of fissions etc.) has been discussed. It has been demonstrated that this method can be insensitive to properties of the zones surrounding the core, whereas is strongly affected by the detector dead time. In turn, the influence of harmonics in the neutron field and of the dispersion of spallation neutrons has proven much less pronounced. PMID- 16381683 TI - Hardening neutron spectrum for advanced actinide transmutation experiments in the ATR. AB - The most effective method for transmuting long-lived isotopes contained in spent nuclear fuel into shorter-lived fission products is in a fast neutron spectrum reactor. In the absence of a fast test reactor in the United States, initial irradiation testing of candidate fuels can be performed in a thermal test reactor that has been modified to produce a test region with a hardened neutron spectrum. Such a test facility, with a spectrum similar but somewhat softer than that of the liquid-metal fast breeder reactor (LMFBR), has been constructed in the INEEL's Advanced Test Reactor (ATR). The radial fission power distribution of the actinide fuel pin, which is an important parameter in fission gas release modelling, needs to be accurately predicted and the hardened neutron spectrum in the ATR and the LMFBR fast neutron spectrum is compared. The comparison analyses in this study are performed using MCWO, a well-developed tool that couples the Monte Carlo transport code MCNP with the isotope depletion and build-up code ORIGEN-2. MCWO analysis yields time-dependent and neutron-spectrum-dependent minor actinide and Pu concentrations and detailed radial fission power profile calculations for a typical fast reactor (LMFBR) neutron spectrum and the hardened neutron spectrum test region in the ATR. The MCWO-calculated results indicate that the cadmium basket used in the advanced fuel test assembly in the ATR can effectively depress the linear heat generation rate in the experimental fuels and harden the neutron spectrum in the test region. PMID- 16381684 TI - Calculation of eigenfunction fluxes in nuclear systems. AB - A new Monte Carlo method is being developed to calculate eigenfunction fluxes in critical or near-critical nuclear systems. The correct estimation of fluxes is essential for radiation protection and shielding near these systems, in addition to isotope production, isotope depletion, nuclear criticality and other applications. The proposed method applies to Monte Carlo criticality eigenvalue calculations in which the fission sites in one generation are used as fission sources in subsequent generations. The usual Monte Carlo power iteration method for such problems often calculates fluxes (eigenfunctions) that are inaccurate and very different in symmetric parts of a problem geometry. The proposed method calculates flux distributions by estimating an approximate fission matrix. The way the fission matrix is estimated and used differs from other recent works. Preliminary results are promising. PMID- 16381685 TI - Application of MCBEND to PBMR shielding analysis. AB - Shielding analysis of an early design of Pebble Bed Modular Reactor (PBMR) has been carried out by using the Monte Carlo code MCBEND. The issues of concern were damage to the core barrel and the reactor pressure vessel (RPV), activation of the core barrel, RPV, top plate and bottom plate, and also burn-up of boron in the control layer underneath the core. The analysis below the core was complicated due to the presence of the de-fuelling chute, which meant that multiplication had to be taken into account. The analysis of boron burn-up was particularly challenging and was tackled using a combination of MCBEND and the criticality code MONK in the depletion mode. The application of MCBEND to the shielding analysis of the PBMR is described, with particular attention being paid to the regions below the core. PMID- 16381686 TI - Evaluation of the neutron spectrum and dose assessment around the venus reactor. AB - An assessment of the neutron field near the VENUS reactor is made in order to evaluate the neutron dose to the operators, particularly in an area near the reactor shielding and in the control room. Therefore, a full MCNPX model of the shielding geometry was developed. The source term used in the simulation is derived from a criticality calculation done beforehand. Calculations are compared to routine neutron dose rate measurements and show good agreement. The MCNPX model developed easily allows core adaptations in order to evaluate the effect of future core configuration on the neutron dose to the operators. PMID- 16381687 TI - Benchmarking of Monte Carlo based shutdown dose rate calculations for applications to JET. AB - The calculation of dose rates after shutdown is an important issue for operating nuclear reactors. A validated computational tool is needed for reliable dose rate calculations. In fusion reactors neutrons induce high levels of radioactivity and presumably high doses. The complex geometries of the devices require the use of sophisticated geometry modelling and computational tools for transport calculations. Simple rule of thumb laws do not always apply well. Two computational procedures have been developed recently and applied to fusion machines. Comparisons between the two methods showed some inherent discrepancies when applied to calculation for the ITER while good agreement was found for a 14 MeV point source neutron benchmark experiment. Further benchmarks were considered necessary to investigate in more detail the reasons for the different results in different cases. In this frame the application to the Joint European Torus JET machine has been considered as a useful benchmark exercise. In a first calculational benchmark with a representative D-T irradiation history of JET the two methods differed by no more than 25%. In another, more realistic benchmark exercise, which is the subject of this paper, the real irradiation history of D-T and D-D campaigns conducted at JET in 1997-98 were used to calculate the shut down doses at different locations, irradiation and decay times. Experimental dose data recorded at JET for the same conditions offer the possibility to check the prediction capability of the calculations and thus show the applicability (and the constraints) of the procedures and data to the rather complex shutdown dose rate analysis of real fusion devices. Calculation results obtained by the two methods are reported below, comparison with experimental results give discrepancies ranging between 2 and 10. The reasons of that can be ascribed to the high uncertainty on the experimental data and the unsatisfactory JET model used in the calculation. A new dedicated JET benchmark experiment will be performed trying to solve these issues. PMID- 16381688 TI - Gamma dose from activation of internal shields in IRIS reactor. AB - The International Reactor Innovative and Secure is a modular pressurised water reactor with an integral design. This means that all the primary system components, such as the steam generators, pumps, pressuriser and control rod drive mechanisms, are located inside the reactor vessel, which requires a large diameter. For the sake of better reliability and safety, it is desirable to achieve the reduction of vessel embrittlement as well as the lowering of the dose beyond the vessel. The former can be easily accomplished by the presence of a wide downcomer, filled with water, which surrounds the core region, while the latter needs the presence of additional internal shields. An optimal shielding configuration is under investigation, for reducing the ex-vessel dose due to activated internals and for limiting the amount of the biological shielding. MCNP 4C calculations were performed to evaluate the neutron and the gamma dose during operation and the 60Co activation of various shields configurations. The gamma dose beyond the vessel from activation of its structural components was estimated in a shutdown condition, with the Monte Carlo code FLUKA 2002 and the MicroShield software. The results of the two codes are in agreement and show that the dose is sufficiently low, even without an additional shield. PMID- 16381689 TI - Measurement and calculation of the fast-neutron and photon spectra from the core boundary to the biological shielding in the WWER-1000 reactor model. AB - The fast-neutron and photon space-energy distributions have been measured in an axially (1.25 m active height) and azimuthally (60 degree symmetry sector) shortened model of the WWER-1000 reactor assembled in the LR-0 experimental reactor. The space-energy distributions have been calculated with the stochastic code MCNP and the deterministic three-dimensional code TORT. Selected results are presented and discussed in the paper. This work has been done in the frame of the EU 5th FW project REDOS REDOS, Reactor Dosimetry: Accurate determination and benchmarking of radiation field parameters, relevant for reactor pressure vessel monitoring. EURATOM Programme, Call 2000/C 294/04). All geometry and material composition data of the model as well as the available experimental data were carefully checked and revised. PMID- 16381690 TI - Radiation safety assessment of a system of small reactors for distributed energy. AB - A passively safe small reactor for a distributed energy system, PSRD, is an integral type of light-water reactor with a thermal output of 100 or 300 MW aimed to be used for supplying district heat, electricity to small grids, and so on. Candidate locations for the PSRD as a distributed energy source are on-ground, deep underground, and in a seaside pit in the vicinity of the energy consumption area. Assessments of the radiation safety of a PSRD were carried out for three cases corresponding to normal operation, shutdown and a hypothetical postulated accident for several siting candidates. Results of the radiation safety assessment indicate that the PSRD design has sufficient shielding performance and capability and that the exposure to the general public is very low in the case of a hypothetical accident. PMID- 16381691 TI - Pressure vessel calculations for VVER-440 reactors. AB - For the determination of the fast neutron load of the reactor pressure vessel a mixed calculational procedure was developed. The procedure was applied to the Unit II of Paks NPP, Hungary. The neutron source on the outer surfaces of the reactor was determined by a core design code, and the neutron transport calculations outside the core were performed by the Monte Carlo code MCNP. The reaction rate in the activation detectors at surveillance positions and at the cavity were calculated and compared with measurements. In most cases, fairly good agreement was found. PMID- 16381693 TI - Modern new nuclear fuel characteristics and radiation protection aspects. AB - The glut of fissile material from reprocessing plants and from the conclusion of the cold war has provided the opportunity to design new fuel types to beneficially dispose of such stocks by generating useful power. Thus, in addition to the normal reactor core complement of enriched uranium fuel assemblies, two other types are available on the world market. These are the ERU (enriched recycled uranium) and the MOX (mixed oxide) fuel assemblies. Framatome ANP produces ERU fuel assemblies by taking feed material from reprocessing facilities and blending this with highly enriched uranium from other sources. MOX fuel assemblies contain plutonium isotopes, thus exploiting the higher neutron yield of the plutonium fission process. This paper describes and evaluates the gamma, spontaneous and alpha reaction neutron source terms of these non-irradiated fuel assembly types by defining their nuclear characteristics. The dose rates which arise from these terms are provided along with an overview of radiation protection aspects for consideration in transporting and delivering such fuel assemblies to power generating utilities. PMID- 16381692 TI - Activation analysis of concrete and graphite in the experimental reactor RUS. AB - The decommissioning and dismantling of nuclear installations after their service life involves the necessary disassembling, handling and disposing of a large amount of radioactive equipment and structures. In particular, the concrete that has been used as a biological reactor shield and graphite that has been used as a moderator-reflector represent the majority of waste, requiring geological disposal. To reduce this undesirable volume to the minimum and to successfully plan the dismantling and disposal of radioactive materials to storage facilities, the activations of the structures should be accurately evaluated. In the framework of the decommissioning and the dismantling of the experimental reactor of the University of Strasbourg, detailed activation estimates have been conducted to characterise the graphite and the structural materials present in the reactor environment. For this purpose, the chemical compositions of fresh graphite samples and different types of concrete have been determined by activation analysis in the research reactors OSIRIS and ORPHEE of CEA Saclay (France). Then, the activations of graphite, concrete and other materials have been calculated in the whole reactor, as a function of the three main nuclear data libraries, i.e. ENDF, JEF and JENDL. In parallel, the activations of representative graphite and concrete samples have been measured experimentally. The comparison of theoretical predictions with experimental values validates the approach and the methodology used in the present study and tests the consistency and the reliability of the nuclear data used for activation analysis. We believe that a similar approach could also be used for the decommissioning of industrial nuclear reactors. PMID- 16381694 TI - Non-destructive method of characterisation of radioactive waste containers using gamma spectroscopy and Monte Carlo techniques. AB - During the decommissioning of the SATURNE accelerator at CEA Saclay (France), a number of concrete containers with radioactive materials of low or very low activity had to be characterised before their final storage. In this paper, a non destructive approach combining gamma ray spectroscopy and Monte Carlo simulations is used in order to characterise massive concrete blocks containing some radioactive waste. The limits and uncertainties of the proposed method are quantified for the source term activity estimates using 137Cs as a tracer element. A series of activity measurements with a few representative waste containers were performed before and after destruction. It has been found that neither was the distribution of radioactive materials homogeneous nor was its density unique, and this became the major source of systematic errors in this study. Nevertheless, we conclude that by combining gamma ray spectroscopy and full scale Monte Carlo simulations one can estimate the source term activity for some tracer elements such as 134Cs, 137Cs, 60Co, etc. The uncertainty of this estimation should not be bigger than a factor of 2-3. PMID- 16381695 TI - SOURCES: a code for calculating (alpha,n), spontaneous fission, and delayed neutron sources and spectra. AB - SOURCES is a computer code that determines neutron production rates and spectra from (alpha,n) reactions, spontaneous fission and delayed neutron emission owing to the decay of radionuclides in homogeneous media, interface problems and three region interface problems. The code is also capable of calculating the neutron production rates due to (alpha,n) reactions induced by a monoenergetic beam of alpha particles incident on a slab of target material. The (alpha,n) spectra are calculated using an assumed isotropic angular distribution in the centre-of-mass system with a library of 107 nuclide decay alpha-particle spectra, 24 sets of measured and/or evaluated (alpha,n) cross sections and product nuclide level branching fractions, and functional alpha particle stopping cross sections for Z < 106. Spontaneous fission sources and spectra are calculated with evaluated half life, spontaneous fission branching and Watt spectrum parameters for 44 actinides. The delayed neutron spectra are taken from an evaluated library of 105 precursors. The code outputs the magnitude and spectra of the resultant neutron sources. It also provides an analysis of the contributions to that source by each nuclide in the problem. PMID- 16381697 TI - Integrated nuclear data utilisation system for innovative reactors. AB - A five-year research and development project on an integrated nuclear data utilisation system was initiated in 2002, for developing innovative nuclear energy systems such as accelerator-driven systems. The integrated nuclear data utilisation system will be constructed as a modular code system, which consists of two sub-systems: the nuclear data search and plotting sub-system, and the nuclear data processing and utilisation sub-system. The system will be operated with a graphical user interface in order to enable easy utilisation through the Internet by both nuclear design engineers and nuclear data evaluators. This paper presents an overview of the integrated nuclear data utilisation system, describes the development of a prototype system to examine the operability of the user interface and discusses specifications of the two sub-systems. PMID- 16381696 TI - An additional performance of HTRS: the waste radiotoxicity minimisation. AB - The management of radioactive waste is a key issue for the present and future use of nuclear energy. In this frame, high temperature reactors (HTRs) have, among others, the capability to burn actinides. After a short introduction on HTRs, the performances of two MC-based burnup codes (Monte Carlo continuous energy burnup and MONTEBURNS) in assessing the ability of these reactors to burn actinides are compared. These codes are necessary for performing ultra-high burnup calculations on HTRs. The best one, in this specific case, results to be MONTEBURNS. It was analysed using HTRs loaded with the following: (1) 1st generation Pu, 600 equivalent full power days; (2) 2nd generation Pu, 645 equivalent full power days; and (iii) 33% 1st generation Pu and 67% Th, 705 equivalent full power days. Finally, it is possible to conclude that HTRs can reduce time when the waste is considered dangerous. Even if the amount of reduction does not solve the whole problem, it represents an important step in the management of radioactive waste. PMID- 16381698 TI - Development of a quantum molecular dynamic (QMD) model to describe fission and fragment production. AB - QMD model coupled with Generalized Evaporation model by S. Furihata (GEM2) is applied for a description of fission nuclei production in p+U interactions at 100 MeV, and for a description of p+Pb --> Bi+X reactions at 10-200 MeV. A good reproduction of the data has been reached. PMID- 16381699 TI - Covariance and sensitivity data generation at ORNL. AB - Covariance data are required to assess uncertainties in design parameters in several nuclear applications. The error estimation of calculated quantities relies on the nuclear data uncertainty information available in the basic nuclear data libraries, such as the US Evaluated Nuclear Data Library, ENDF/B. The uncertainty files in the ENDF/B library are obtained from the analysis of experimental data and are stored as variance and covariance data. In this paper we address the generation of covariance data in the resonance region done with the computer code SAMMY. SAMMY is used in the evaluation of the experimental data in the resolved and unresolved resonance energy regions. The data fitting of cross sections is based on the generalised least-squares formalism (Bayesian theory) together with the resonance formalism described by R-matrix theory. Two approaches are used in SAMMY for the generation of resonance parameter covariance data. In the evaluation process SAMMY generates a set of resonance parameters that fit the data, and, it provides the resonance parameter covariances. For resonance parameter evaluations where there are no resonance parameter covariance data available, the alternative is to use an approach called the 'retroactive' resonance parameter covariance generation. In this paper, we describe the application of the retroactive covariance generation approach for the gadolinium isotopes. PMID- 16381700 TI - Measurement of neutron inelastic scattering cross sections for 52Cr from threshold up to 18 MeV. AB - The powerful white neutron spectrum from Gelina is used for the investigation of gamma ray production cross sections in the reaction 52Cr(n,n'gamma)52Cr. The gamma rays from the inelastic process are detected with two large volume HPGe detectors placed at 110 degrees and 150 degrees with respect to the beam direction. The neutron flux has been measured with a 235U fission chamber. For the main transitions the inelastic cross section is given with an incident neutron energy resolution ranging from 1.12 keV at 1 MeV to 35.7 keV at 10 MeV, the overall statistical error being <5%. Based on the known 52Cr level scheme, the level and total inelastic cross sections have been obtained up to 3.77 MeV excitation energy. Progress on the implementation of a new acquisition system based on a fast digitiser is presented in this paper. PMID- 16381701 TI - Neodymium neutron cross section measurements. AB - Neutron capture and transmission measurements were performed by the time-of flight technique at the Rensselaer Polytechnic Institute LINAC using metallic neodymium samples. The capture measurements were made at the 25-m-long flight station with a 16-segment NaI(Tl) multiplicity detector, and the transmission measurements were performed at 15 and 25 m flight stations with a 6Li glass scintillation detector. After the data were collected and reduced, resonance parameters were determined by simultaneously fitting the transmission and capture data with the multilevel R-matrix Bayesian code SAMMY. The resonance parameters for all naturally occurring neodymium isotopes lie within the energy range of 1.0 500 eV. The resulting resonance parameters were used to calculate the capture resonance integral with this energy region and were compared to calculations obtained when using the resonance parameters from ENDF-B/VI. The RPI parameters gave a resonance integral value of 32 +/- 0.5 b that is approximately 7% lower than that obtained with the ENDF-B/VI parameters. The current measurements significantly reduce the statistical uncertainties on the resonance parameters when compared with previously published parameters. PMID- 16381702 TI - Neutron shielding calculations in a proton therapy facility based on Monte Carlo simulations and analytical models: criterion for selecting the method of choice. AB - Proton therapy facilities are shielded to limit the amount of secondary radiation to which patients, occupational workers and members of the general public are exposed. The most commonly applied shielding design methods for proton therapy facilities comprise semi-empirical and analytical methods to estimate the neutron dose equivalent. This study compares the results of these methods with a detailed simulation of a proton therapy facility by using the Monte Carlo technique. A comparison of neutron dose equivalent values predicted by the various methods reveals the superior accuracy of the Monte Carlo predictions in locations where the calculations converge. However, the reliability of the overall shielding design increases if simulation results, for which solutions have not converged, e.g. owing to too few particle histories, can be excluded, and deterministic models are being used at these locations. Criteria to accept or reject Monte Carlo calculations in such complex structures are not well understood. An optimum rejection criterion would allow all converging solutions of Monte Carlo simulation to be taken into account, and reject all solutions with uncertainties larger than the design safety margins. In this study, the optimum rejection criterion of 10% was found. The mean ratio was 26, 62% of all receptor locations showed a ratio between 0.9 and 10, and 92% were between 1 and 100. PMID- 16381703 TI - Neutron radiation area monitoring system for proton therapy facilities. AB - A neutron radiation area monitoring system has been developed for proton accelerator facilities dedicated to cancer therapy. The system comprises commercial measurement equipment, computer hardware and a suite of software applications that were developed specifically for use in a medical accelerator environment. The system is designed to record and display the neutron dose equivalent readings from 16 to 24 locations (depending on the size of the proton therapy centre) throughout the facility. Additional software applications provide for convenient data analysis, plotting, radiation protection reporting, and system maintenance and administration tasks. The system performs with a mean time between failures of >6 months. Required data storage capabilities and application execution times are met with inexpensive off-the-shelf computer hardware. PMID- 16381704 TI - Patient neutron dose equivalent exposures outside of the proton therapy treatment field. AB - A large fraction of dose to healthy tissue located outside of the treatment field during proton therapy is attributable to neutrons produced in the beam-delivery apparatus. In this work, the neutron dose equivalent (H) per therapeutic proton absorbed dose (D) was estimated for typical treatment conditions as a function of range modulation width, angle with respect to the incident proton beam, and the distance from the isocentre at the Harvard Cyclotron Laboratory's (Cambridge, MA) passively spread treatment field using Monte Carlo simulations. For a beam with 16 cm penetration (depth) and a 5 x 5 cm2 lateral field size at the patient location along the incident beam direction at 100 cm from the isocentre, the predicted H/D values are 0.35 and 0.60 mSv Gy(-1) from the simulations and measurements, respectively. At all locations, the predicted H/D values are within a factor of 2 and 3 of the measured result for no modulation and 8.2 cm of modulation, respectively. PMID- 16381705 TI - Virtual commissioning of a treatment planning system for proton therapy of ocular cancers. AB - The virtual commissioning of a treatment planning system (TPS) for ocular proton beam therapy was performed using Monte Carlo (MC) simulations and a model of a double-scattering ocular treatment nozzle. The simulations produced both the input data required by the TPS and the dose distributions to validate the analytical predictions from the TPS. An MC simulation of a typical ocular melanoma treatment was compared with the TPS predictions, revealing generally good agreement in the absorbed dose distribution. However, in the depth-dose profiles, differences >5% existed in the proximal region of all validation cases considered. Comparison of the radiation coverage at or above the 90% dose level, showed that MC calculated coverage was 82% and 68% of the coverage calculated by the TPS in two planes intersecting the tumour. PMID- 16381706 TI - Therapeutic step and shoot proton beam spot-scanning with a multi-leaf collimator: a Monte Carlo study. AB - In step and shoot spot-scanning, a small-diameter proton beam is magnetically swept and varied in energy in order to cover the tumour. Initial estimates of the beam size indicate that additional collimating hardware will be needed for lower energy proton beams in order to achieve a clinically acceptable lateral dose falloff at the edge of the proton beam. In this report, we present dosimetric data from Monte Carlo simulations with a model of a simple multileaf collimator which indicate that such a device may be used to improve the lateral dose falloff. The dosimetric quantities relevant to the clinical usefulness of the device are studied, including lateral penumbra, leaf transmission and scalloping effect. Multileaf collimation is compared with a differential spot-weighting technique of sharpening the lateral dose falloff. PMID- 16381707 TI - Target station shielding issues at the spallation neutron source. AB - Recent spallation neutron source shielding activities in support of the neutron beam shutters and the hot cell walls are presented. Existing neutron beam shutters can be replaced with concrete at low power or with concrete and steel at approximately 500 kW of beam power. Potential voids in the hot cell walls are analysed to determine the impact on dose rates as a function of void size. A change in the type of shielding work is noted as the project moved from the early design stages as a 'green field' site to the current stage as a construction project nearing completion, where issues to be addressed are approaching retrofit type analyses. PMID- 16381708 TI - Calculations for the availability of photoneutron using synchrotron radiation. AB - The availability of the neutrons due to photonuclear reactions has been discussed by using synchrotron radiation with the beryllium targets. The superconducting wiggler with the magnetic field of approximately 10 T, which is installed into an 8 GeV class storage ring, can emit intense and high-energy photons to produce neutrons. By using MCNPX, the simulations were performed for the conceptual design of the neutron beamline to estimate the available intensity and to investigate the shield conditions. The results were discussed in comparison with other research reactors. PMID- 16381709 TI - Shielding design of an underground experimental area at point 5 of the CERN Super Proton Synchrotron (SPS). AB - Increasing projected values of the circulating beam intensity in the Super Proton Synchrotron (SPS) and decreasing limits to radiation exposure, taken with the increasing non-acceptance of unjustified and unoptimised radiation exposures, have led to the need to re-assess the shielding between the ECX and ECA5 underground experimental areas of the SPS. Twenty years ago, these experimental areas at SPS-Point 5 housed the UA1 experiment, where Carlo Rubbia and his team verified the existence of W and Z bosons. The study reported here describes such a re-assessment based on simulations using the multi-purpose FLUKA radiation transport code. This study concludes that while the main shield which is made of concrete blocks and is 4.8 m thick satisfactorily meets the current design limits even at the highest intensities presently planned for the SPS, dose rates calculated for liaison areas on both sides of the main shield significantly exceed the design limits. Possible ways of improving the shielding situation are discussed. PMID- 16381710 TI - The TRADE experiment: shielding calculations for the building hosting the subcritical system. AB - The TRADE project (TRiga Accelerator Driven Experiment), to be performed at the existing TRIGA reactor at ENEA Casaccia, has been proposed as a validation of the accelerator-driven system (ADS) concept. TRADE will be the first experiment in which the three main components of an ADS--the accelerator, spallation target and sub-critical blanket--are coupled at a power level sufficient to encounter reactivity feedback effects. As such, TRADE represents the necessary intermediate step in the development of hybrid transmutation systems, its expected outcomes being considered crucial--in terms of proof of stability of operation, dynamic behaviour and licensing issues--for the subsequent realisation of an ADS Transmutation Demonstrator. An essential role in the feasibility study of the experiment is played by radioprotection calculations. Such a system exhibits new characteristics with respect to a traditional reactor, owing to the presence of the proton accelerator. As beam losses always occur under normal operating conditions of an accelerator, shielding studies need to be performed not only around the reactor but also along the beam line from the accelerator to the spallation target. This paper illustrates a preliminary evaluation, using Monte Carlo methods, of the additional shielding to be located around the reactor structures, the beam transport line and the existing reactor building to allow access into the reactor hall and to restrict the doses outside to their legal limits. PMID- 16381711 TI - Shielding variation effects for 250 MeV protons on tissue targets. AB - This paper provides results of computer simulation studies with the goal to analyse issues regarding radiation protection for personnel, patients and third persons involved in hadron therapy treatment. The treatment room and the patient are modelled by simple cylindrical geometries at incident proton energies of 250 MeV. Monte Carlo simulations of the energy and angular dependence of proton, neutron and photon radiation fields and resulting ambient dose equivalent distributions outside the shielding walls are performed. In order to investigate systematic uncertainties due to the shielding materials and inherent to the computer models, various concrete compositions, densities and water contents are modelled, and the influence of simulation parameters on the results obtained is determined. Generally, good agreement is found between results provided by MCNPX and FLUKA computer codes. Variations in neutron ambient dose attenuation from -50 to +/-30% are found due to varying concrete composition. Changes in the water content of the concrete in the order of 8% may cause variations up to 20%. PMID- 16381712 TI - Calculations of neutron and photon source terms and attenuation profiles for the generic design of the SPEAR3 storage ring shield. AB - The FLUKA Monte Carlo particle generation and transport code was used to calculate shielding requirements for the 3 GeV, 500 mA SPEAR3 storage ring at the Stanford Synchrotron Radiation Laboratory. The photon and neutron dose equivalent source term data were simulated for a 3 GeV electron beam interacting with two typical target/shielding geometries in the ring. The targets simulated are a rectangular block of 0.7 cm thick copper and a 5 cm thick iron block, both tilted at 1 degree relative to the beam direction. Attenuation profiles for neutrons and photons in concrete and lead as a function of angle at different shield thicknesses were calculated. The first, second and equilibrium attenuation lengths of photons and neutrons in the shield materials are derived from the attenuation profiles. The source term data and the attenuation lengths were then used to evaluate the shielding requirements for the ratchet walls of all front ends of the SPEAR3 storage ring. PMID- 16381713 TI - Shielding for the upgraded Duke Free Electron Laser Laboratory. AB - The Duke FEL Laboratory is undergoing a series of upgrades staggered over time that will greatly increase the capabilities of the machines and by the same token the importance of radiation safety issues. In this paper, we present the scope of the planned upgrades and provide several specific examples of shielding calculations. We also present our effort to correlate calculations with experimental measurements. PMID- 16381714 TI - Monte Carlo simulations for the shielding of the future high-intensity accelerator facility FAIR at GSI. AB - The Gesellschaft fur Schwerionenforschung (GSI) is planning a significant expansion of its accelerator facilities. Compared to the present GSI facility, a factor of 100 in primary beam intensities and up to a factor of 10,000 in secondary radioactive beam intensities are key technical goals of the proposal. The second branch of the so-called Facility for Antiproton and Ion Research (FAIR) is the production of antiprotons and their storage in rings and traps. The facility will provide beam energies a factor of approximately 15 higher than presently available at the GSI for all ions, from protons to uranium. The shielding design of the synchrotron SIS 100/300 is shown exemplarily by using Monte Carlo calculations with the FLUKA code. The experimental area serving the investigation of compressed baryonic matter is analysed in the same way. In addition, a dose comparison is made for an experimental area operated with medium energy heavy-ion beams. Here, Monte Carlo calculations are performed by using either heavy-ion primary particles or proton beams with intensities scaled by the mass number of the corresponding heavy-ion beam. PMID- 16381715 TI - The refined shielding design for the cyclotron room of the Buddhist Tzu Chi General Hospital. AB - Full-scale Monte Carlo simulations of the cyclotron room of the Buddhist Tzu Chi General Hospital were carried out to improve the inadequate maze design. The double differential neutron source from the 18O(p,n)18F reaction was adopted for the calculation. The weight window variance reduction technique, where the weight window was set by applying the adjoint flux, has been implemented in the MCNP run to facilitate the calculation of the dose rates outside the cyclotron room. Dose rates including neutron and gamma-ray components were calculated for some maze shielding modifications. PMID- 16381716 TI - Beam dumps design and local radiation protection at TERA synchrotron. AB - The realisation of the National Center of Hadrontherapy was funded by the Italian Government in 2002. The Centre will be built in the area of Pavia (Italy). The synchrotron designed in the framework of this programme will accelerate protons and carbon ions up to 250 MeV and 400 MeV u(-1), respectively. Some of the main aspects which were taken into account in the design of the acceleration system are the patient's safety and the beam control. From this point of view an important role is played by the beam dumps in the synchrotron ring and upstream of the extraction system. In particular, an horizontal and a vertical beam dump will be installed in the synchrotron ring: the former will be used for lowering the beam intensity and the latter for beam abortion. The dump at the extraction will absorb the particles during the mounting and the falling ramps of the synchrotron magnetic cycle, thus extracting only the flat top of the ion spill. Beam dumps can produce intense fields of secondary radiation (neutrons, charged light-hadrons and photons) and high rates of induced activity, since they can absorb the beam completely. Usually they have to be shielded to protect the electronics during machine operation and to attenuate the radiation dose below the limits imposed by the law when the personnel access to the synchrotron hall. The part of the shielding design of the beam dumps concerning with the acceleration of protons was made using Monte Carlo simulations with the FLUKA code. Both induced activity and secondary radiation were taken into account. The shields against secondary radiation produced by carbon ions were designed, referring only to secondary neutrons, taking double-differential distributions from the literature as sources for the FLUKA simulations. The induced activity from carbon ions interactions was estimated analytically, using the data generated by the EPAX 2 code. The dose-equivalent rates from the induced radionuclides were calculated at 1 m from the shielded dumps, taking into account the contribution of activated components of the synchrotron ring. PMID- 16381717 TI - Evaluation of silicon neutron resonance parameters in the thermal to 1800 keV energy range. AB - Because silicon is a major constituent of concrete and soil, neutron and gamma ray information on silicon is important for reactor shielding and criticality safety calculations. Therefore, much effort was put into the ENDF/B-VI evaluation for the three stable isotopes of silicon. The neutron capture cross section of natural silicon was recently measured at the Oak Ridge Electron Linear Accelerator (ORELA) in the energy range 1-700 keV. Using the ENDF/B-VI evaluation for initial values, a new evaluation of the resonance parameters was performed by adding the results of the ORELA capture measurements to the experimental database. The computer code SAMMY was used for the analysis of the experimental data; the new version of SAMMY allows accurate calculations of the self-shielding and multiple scattering effects in the capture measurements. The accuracy of the radiative capture widths of the resonances was improved by this analysis. Accurate values of the s-, p- and d-wave neutron strength functions were also obtained. Although the resonance capture component of the present evaluation is 2 3 times smaller than that in ENDF/B-VI, the total capture cross section is much larger, at least for energies >250 keV, because the direct capture component contributes values of the same order of magnitude as the resonance component. The direct component was not taken into account in the ENDF/B-VI evaluation and was calculated for the first time in the present evaluation. PMID- 16381718 TI - Generation and validation of a shielding library based on ENDF/B-VI.8. AB - For various shielding and reactor pressure vessel dosimetry applications, a pseudo-problem-independent neutron-photon coupled MATXS-format library, based on the last release of ENDF/B-VI, has been generated as a part of the update program for KASHIL-E6, which was based on ENDF/B-VI.5. It has a VITAMIN-B6 neutron and photon energy group structures, i.e. 199 groups for the neutron and 42 groups for the photon. The neutron and photon weighting functions and the Legendre orders of scattering are the same as in KASHIL-E6. The library has been validated through some benchmarks: the PCA-REPLICA and NESDIP-2 experiments for the light-water reactor (LWR) pressure vessel facility benchmark, the Winfrith Iron 88 experiment for the validation of the iron data and the Winfrith Graphite experiment for the validation of the graphite data. These calculations were performed by the TRANSX/DANTSYS code system. In addition, the substitutions of the JENDL-3.3 and JEFF-3.0 data for Fe, Cr, Cu and Ni, which are very important nuclides for shielding analyses, were investigated to estimate the effects on the benchmark calculation results. PMID- 16381719 TI - MENDL2 and IEAF2001 nuclide production yields databases verification in inter medium energy range. AB - This work presents results of computer simulation of two experiments which aim at measuring the threshold activation reaction rates in 12C, 19F, 27Al, 59Co, 63Cu, 65Cu, 64Zn, 93Nb, 115In, 169Tm, 181Ta, 197Au, and thin samples placed inside and outside a 0.8 GeV proton-irradiated 4-cm thick W target and a 92-cm-thick W-Na composite target of 15 cm diameter both. In total, more than 1000 values of activation reaction were determined in both the experiments. The measured reaction rates were compared with the rates simulated by the LAHET code with the use of several nuclear databases for the respective excitation functions, namely, MENDL2 together with MENDL2P for cross sections of protons and neutrons up to 100 MeV, and with the recently developed IEAF-2001 that provides neutron cross sections up to 150 MeV. A comparison of simulation-to-experiment agreement obtained with MENDL2 and IEAF-2001 is presented. The agreement between simulation and experiment has been found general satisfactory for both databases. However, further studies should be done to improve the simulation of production of secondary protons and high-energy neutrons, as well as the high-energy neutron elastic scattering. Our results allow to conclude about the reliability of the transport codes and databases used to simulate the accelerator driven systems (ADS), particularly with Na-cooled W targets. High-energy threshold excitation functions to be used in the activation-based unfolding of neutron spectra inside the ADS can also be inferred from these results. PMID- 16381720 TI - Impact of high-energy nuclear data on radioprotection in spallation sources. AB - The high-energy programme of the HINDAS European project has provided a large amount of experimental data and led to a better understanding of the spallation reaction mechanism and the development of more reliable spallation models. These data, or the new models, which have been implemented into high-energy transport codes, can be now used to predict with a larger confidence or, at least with a known uncertainty, some important quantities for the design of spallation sources. In this paper, examples concerning the residue production in a Pb-Bi target and the high-energy neutrons escaping the target are presented. In the first case, the activity and the amount of radioactive volatile elements that can be released, in case of a containment failure, are calculated and the level of confidence of the calculation is assessed. The second example shows that the models correctly predict the high-energy tail of the neutron spectrum, which is important for radioprotection in the facility. PMID- 16381721 TI - Air kerma rate constants for gamma emitters used most often in practice. AB - It is often required to estimate the dose rate at a distance from radionuclides that are sources of X rays and gamma rays. Such calculations may be required for planning radiation protection measures in the vicinity of radioactive sources or patients containing radionuclides, calibrations of radiation instruments or for estimating the absorbed dose rate to patients receiving brachytherapy. The factor relating activity and air kerma rate is called air kerma rate constant- gamma(delta). In this paper, the results of recalculation of this quantity for unfiltered point sources of radionuclides in practice used most often are given. The calculations included corrections for internal conversion of X rays and gamma rays and detailed accounting of the generation of the K and L series X rays from internal conversion and electron capture. Particular air kerma rate constants were calculated for each discrete line in the photon spectrum of radionuclide with a yield per decay event >0.01% and the energy >20 keV. Since the energy structure of the photon spectra and accessible discrete numerical values of the mass energy-transfer coefficient for air are not the same, the cubic spline interpolation was used to obtained the coefficient, where the photon spectrum data are available. In the calculation, the latest gamma ray spectral data for all radionuclides and latest data for the mass energy-transfer coefficient for air are used. Air kerma rate constants for the following 35 radionuclides are calculated: 11C, 13N, 15O, 18F, 24Na, 42K, 43K, 51Cr, 52Fe, 59Fe, 57Co, 58Co, 60Co, 67Ga, 68Ga, 75Se, 99Mo, 99mTc, 111In, 113mIn, 123I, 125I, 131I, 127Xe, 133Xe, 137Cs, 152Eu, 154Eu, 170Tm, 182Ta, 192Ir, 197Hg, 198Au, 201Tl and 241Am. PMID- 16381722 TI - Radiation protection and shielding design--strengthening the link. AB - The improvement in quality and flexibility of shielding methods and data has been progressive and beneficial in opening up new opportunities for optimising radiation protection in design. The paper describes how these opportunities can best be seized by taking a holistic view of radiation protection, with shielding design being an important component part. This view is best achieved by enhancing the role of 'shielding assessors' so that they truly become 'radiation protection designers'. The increase in speed and efficiency of shielding calculations has been enormous over the past decades. This has raised the issue of how the assessor's time now can be best utilised; pursuing ever greater precision and accuracy in shielding/dose assessments, or improving the contribution that shielding assessment makes to radiological protection and cost-effective design. It is argued in this paper that the latter option is of great importance and will give considerable benefits. Shielding design needs to form part of a larger radiation protection perspective based on a deep understanding/appreciation of the opportunities and constraints of operators and designers, enabling minimal design iterations, cost optimisation of alternative designs (with a 'lifetime' perspective) and improved realisation of design intent in operations. The future of shielding design development is argued to be not in improving the 'toolkit', but in enhanced understanding of the 'product' and the 'process' for achieving it. The holistic processes being developed in BNFL to realise these benefits are described in the paper and will be illustrated by case studies. PMID- 16381723 TI - Comparison of deterministic and Monte Carlo methods in shielding design. AB - In shielding calculation, deterministic methods have some advantages and also some disadvantages relative to other kind of codes, such as Monte Carlo. The main advantage is the short computer time needed to find solutions while the disadvantages are related to the often-used build-up factor that is extrapolated from high to low energies or with unknown geometrical conditions, which can lead to significant errors in shielding results. The aim of this work is to investigate how good are some deterministic methods to calculating low-energy shielding, using attenuation coefficients and build-up factor corrections. Commercial software MicroShield 5.05 has been used as the deterministic code while MCNP has been used as the Monte Carlo code. Point and cylindrical sources with slab shield have been defined allowing comparison between the capability of both Monte Carlo and deterministic methods in a day-by-day shielding calculation using sensitivity analysis of significant parameters, such as energy and geometrical conditions. PMID- 16381724 TI - Neutron shielding material based on colemanite and epoxy resin. AB - In recent years, there has been a need for compact shielding design such as self shielding of a PET cyclotron or upgradation of radiation machinery in existing facilities. In these cases, high performance shielding materials are needed. Concrete or polyethylene have been used for a neutron shield. However, for compact shielding, they fall short in terms of performance or durability. Therefore, a new type of neutron shielding material based on epoxy resin and colemanite has been developed. Slab attenuation experiments up to 40 cm for the new shielding material were carried out using a 252Cf neutron source. Measurement was carried out using a REM-counter, and compared with calculation. The results show that the shielding performance is better than concrete and polyethylene mixed with 10 wt% boron oxide. From the result, we confirmed that the performance of the new material is suitable for practical use. PMID- 16381725 TI - Shielding design for the PANDA spectrometer at the Munich high-flux reactor FRM II. AB - The start-up of the Munich high-flux reactor FRM-II is in progress on. At the beam tube SR-2 the spectrometer PANDA has been installed. It is at three-axis neutron spectrometer looking onto a slightly under-moderated cold neutron source. For polarisation analysis, PANDA is equipped with a vertical cryomagnet producing fields up to 14.5 T for the sample. To get an appropriate shielding of the high intensity instrument, one has to take into account the large cross section of the primary beam, several restrictions using magnetic materials, limitations in loading the site and finally, has to keep the lateral extent of the shielding small to allow for high-scattering angles. The shielding has been designed on the basis of the results, which were achieved by the combined use of both the Monte Carlo code MCNP-4B2 and an analytical method based on one-dimensional dose transmission functions. PMID- 16381726 TI - Monte Carlo evaluations and the source normalisation problem. AB - It is well known that Transport Theory calculations give results which are immediately intelligible, as they take into account the source explicitly. Monte Carlo evaluations on the contrary are given per source particle, and they have to be normalized according to the actual source to be completely intelligible. Unfortunately in MC evaluations sometimes one can be led, to reach good results from a statistic point of view (that is low variance values), to manipulate somehow the source instead of sharply define it, in order either to fully exploit the features of his own code or to bypass and simplify actual physical difficulties, but on the other hand this procedure strongly increases the complexity of the source normalization process, which becomes (or can become) the true problem indeed. Two examples will be analyzed: a shielding problem of a transportation cask which will be investigated by means of MCNP, and a shielding problem of a proton beam which will be investigated by means of FLUKA. In both the problems a correct, sharply defined description of the source leads to hardly manageable situations, when "tricky" adjustments on the source instead lead smoothly to statistically good results while the source normalizations are anyway solved without too much complexity. PMID- 16381727 TI - Design of radiation shielding for the proton therapy facility at the National Cancer Center in Korea. AB - The design of radiation shielding was evaluated for a proton therapy facility being established at the National Cancer Center in Korea. The proton beam energy from a 230 MeV cyclotron is varied for therapy using a graphite target. This energy variation process produces high radiation and thus thick shielding walls surround the region. The evaluation was first carried out using analytical expressions at selected locations. Further detailed evaluations have been performed using the Monte Carlo method. Dose equivalent values were calculated to be compared with analytical results. The analytical method generally yielded more conservative values. With consideration of adequate occupancy factors annual dose equivalent rates are kept <1 mSv y(-1) in all areas. Construction of the building is expected to be completed near the end of 2004 and the installation of therapy equipment will begin a few months later. PMID- 16381728 TI - Monte Carlo modelling of a simple accident dosemeter. AB - A simple dosemeter made of a sulphur tablet, bare and cadmium-covered indium foils and a cadmium-covered copper foil has been modelled using MCNP5. Studies of the model without phantom or other confounding factors have shown that the cross sections and fluence-to-dose factors generated by the Monte Carlo method agree with those generated by analytic expressions for the high energy component. In this study, a comparison of the effect of location on phantoms and an extension to low and intermediate energies is done. The activities expected from exposure to four critical assemblies on phantom is calculated and compared with observations. PMID- 16381729 TI - Radiation protection system at the RIKEN RI beam factory. AB - The RIKEN RI (radioactive isotope) Beam Factory is scheduled to commence operations in 2006, and its maximum energy will be 400 MeV u(-1) for ions lighter than Ar and 350 MeV u(-1) for uranium. The beam intensity will be 1 pmicroA (6 x 10(12) particles s(-1)) for any element at the goal. For the hands-on-maintenance and the rational shield thickness of the building, the beam loss must be controlled with several kinds of monitors. Three types of radiation monitors will be installed. The first one consists of a neutron dose equivalent monitor and an ionisation chamber, which are commercially available area monitors. The second one is a conventional hand-held dose equivalent monitor wherein the logarithmic signal is read by a programmable logic controller based on the radiation safety interlock system (HIS). The third one is a simple plastic scintillator called a beam loss monitor. All the monitors have threshold levels for alarm and beam stop, and HIS reads all these signals. PMID- 16381730 TI - Development of lung and soft tissue substitutes for photons. AB - The use of solid tissue substitutes is a well-accepted and common practice in dosimetric studies and in the production of counting standards for radiological protection. However, only a few solid tissue substitutes simulating a particular body tissue with respect to a set of physical characteristics are commercially available. Hence, we have developed polyurethane-based tissue substitutes simulating soft tissue, muscle, muscle-adipose mixture tissue (90% muscle + 10% adipose), brain, cartilage, larynx, thyroid, trachea, liver, kidney, skin and lungs. Tissue substitutes for photons were formulated using the basic data method together with an equation for calculating the optimum relative mass of corrective additives. The tissue substitutes were formulated to be phantom materials in the photon energy range of at least 8 keV-10 MeV. In particular, they were designed to match the body tissues with linear attenuation coefficients for low photon energy (13.6, 17.2 and 20.2 keV from 239Pu) and to have the same mass densities as the tissues. The tissue substitutes developed in the present study were examined for the photon transmissions using 16.6 keV KX rays from 93Nb(m). The experimental transmission curves of the tissue substitutes were found to be consistent with those derived from data on the body tissues in ICRP Publication 23. It was found that the developed tissue substitutes are suitable to the corresponding body tissues defined by ICRP. PMID- 16381731 TI - Effective dose for patients undergoing coronary and femoral intravascular radiotherapy involving an HDR 192Ir source. AB - Effective dose equivalent (EDE), and effective dose (ED) for coronary and femoral Intravascular brachytherapy (IVBT) procedures involving a 370 GBq (10 Ci) HDR 192Ir gamma source are tabulated. MIRD stylised models and the MCNP Monte Carlo code were used for the calculations. For coronary irradiation, the normalised EDE is 0.18 mSv (GBq min)(-1) and the ED is 0.056 mSv (GBq min)(-1). For femoral IVBT, the normalised EDE is 0.01629 mSv (GBq min)(-1) and the ED is 0.01195 mSv (GBq min)(-1). Although the medical benefits to a patient undergoing IVBT are often significant and justified, patient doses are high compared with dose limits for radiation protection purposes. As IVBT is becoming a routine procedure, data in this paper could be useful to manage the procedures efficiently. PMID- 16381732 TI - ALARA pre-job studies using the VISIPLAN 3D ALARA planning tool. AB - The optimisation of the radiation protection for the workers in nuclear industry is an important part of the safety culture. The application of the ALARA concept (to keep exposures as low as reasonably achievable) is not always straightforward as it is influenced by the site geometry, source distribution and work organisation. A good ALARA pre-job study must therefore be performed and should contain predicted doses for the different suggested work scenarios and provide a quantitative basis to select between various alternative work scenarios for a specific operation. In order to handle this information, SCK-CEN developed the VISIPLAN 3D ALARA planning tool. The tool makes it possible to evaluate the dose due to external gamma exposure based on the simulation of work scenarios taking into account worker positions and subsequent geometry and source distribution changes in a three-dimensional environment. PMID- 16381733 TI - Bremsstrahlung doses from natural uranium ingots. AB - In the past, some privately owned commercial facilities in the United States were involved in producing or processing radioactive materials used in the production of atomic weapons. Seven different geometrical objects, representative of the configurations of natural uranium metal potentially encountered by workers at these facilities, are modelled to determine gamma ray and bremsstrahlung dose rates. The dose rates are calculated using the MCNP5 code and also by using the MICROSHIELD point-kernel code. Both gamma ray and bremsstrahlung dose rates are calculated and combined to obtain a total dose rate. The two methods were found to be in good agreement despite differences in modelling assumptions and method differences. Computed total dose rates on the surface of these objects ranged from approximately 51-84 microSv h(-1) and 17-95 microSv h(-1) using the MCNP5 and the MICROSHIELD modeling, respectively. The partitioning of the computed dose rates between gamma rays and bremsstrahlung were the same order of magnitude for each object. PMID- 16381734 TI - Radiation protection potential of MOX-fuel doped with 231Pa and Cs radioisotopes. AB - The paper addresses the problem of MOX-fuel self-protection during full cycle of MOX-fuel management. Under conditions of the closed LWR cycle the proliferation resistance levels were evaluated for fresh and spent MOX-fuel with 231Pa and Cs feed. As it follows from the paper results, combination of these two admixtures being doped into MOX-fuel is able to enhance the inherent radiation barrier and to weaken shortcomings of both proliferation deterrents. PMID- 16381735 TI - Modelling of a simple bunker problem with Monte Carlo codes TRIPOLI 4.3 and MCNPX 2.4 to test the efficiency of the biasing methods. PMID- 16381736 TI - The radon inverse dose rate effect and high-LET galactic hazards. AB - The lung dose rate per unit 222Rn concentration in enclosed spaces is shown to experience transitions at high radon concentrations. This has implications on the radon inverse dose rate effect. At an air change rate (ACH) of 0.194 h(-1) and relative humidity (RH) of 52.3% in a 0.283 m3 test chamber, the total human lung dose for an adult male in a residential setting (breathing rate 0.78 m3 h(-1)) would undergo a reduction of 2.5 using the ICRP 66 human respiratory tract model and the BEIR VI methodology. Using the same methodology of both Cross (Pacific Northwest Laboratory rat exposures) and Lubin et al. (miners dose rates), adjustments are necessary for effects of RH and ACHs. These adjustments, however, do not affect the reduction behaviour. It is thus shown that the enhanced deposition effect (EDE) must influence the magnitude of the purported inverse dose rate effect (IDRE). In the analysis of animal data, Cross rat exposures in a 2.0 m3 chamber, a reduction in lung dose is estimated to be over a factor of 3 the transition between the 50 and 500 WLM week(-1) dose rate range. For an estimation of the EDE, using a hypothetical 30 m3 enclosure for underground miners, we obtain a factor of approximately 4 in human lung dose reduction. Although the extensive analyses required make these results qualitative, the EDE behaviour is sufficiently conclusive that these estimates show that the radon IDRE for lung cancer must be an EDE dosimetric issue as well as a radiological lung cell dose response issue. The consequence of analysis of other animal data would achieve the same conclusion. PMID- 16381737 TI - A new human eye model for ophthalmic brachytherapy dosimetry. AB - The present work proposes a new mathematical eye model for ophthalmic brachytherapy dosimetry. This new model includes detailed description of internal structures that were not treated in previous works, allowing dose determination in different regions of the eye for a more adequate clinical analysis. Dose calculations were determined with the MCNP-4C Monte Carlo particle transport code running n parallel environment using PVM. The Amersham CKA4 ophthalmic applicator has been chosen and the depth dose distribution has been determined and compared to those provide by the manufacturer. The results have shown excellent agreement. Besides, absorbed dose values due to both 125I seeds and 60Co plaques were obtained for each one of the different structures which compose the eye model and can give relevant information in eventual clinical analyses. PMID- 16381738 TI - Monte Carlo modelling of a voxel head phantom for in vivo measurement of bone seeker nuclides. AB - Whole-body counters (WBCs) are used for the assessment of the internal contamination of actinides in the human body. WBCs require adequate calibration procedures that rely on the use of suitable calibration phantoms. A previous study carried out at the ENEA-Radiation Protection Institute was aimed at designing a head calibration phantom in which a heterogeneous distribution of 241Am point sources could satisfactorily approximate an assumed homogeneous contamination throughout the head bones. Suitable correction factors for the WBC detection efficiencies were evaluated with Monte Carlo. The present paper summarises the main aspects and implications of an advanced modelling technique based on a VOXEL approach. The methodology could be extended to other bone-seeker radionuclides. PMID- 16381739 TI - French comparison exercise with the rotating neutron spectrometer, 'ROSPEC'. AB - The French laboratories in charge of 'neutron' dosimetry using the spectrometer 'ROSPEC', formed a working group in 2001. The participants began to study the behaviour of the instrument with a comparison exercise in broad energy neutron fields recommended by the International Organisation for Standardisation (ISO) and available at the LMDN in Cadarache. The complete version of the ROSPEC is made up of six spherical proportional counters fixed to a rotating platform. These counters cover different energy ranges which overlap each other to provide a link between the detectors, within the energy range from thermal neutrons to 4.5 MeV. The irradiation configurations chosen were ISO standard sources (252Cf, (252Cf+D2O)(/Cd), 241Am-Be) and the SIGMA facility. The results show that the 'thermal and epithermal' neutron fluence was widely overestimated by the spectrometer in all configurations. PMID- 16381740 TI - Analysis of neutron and photon response of a TLD-ALBEDO personal dosemeter on an ISO slab phantom using TRIPOLI-4.3 Monte Carlo code. AB - TRIPOLI-4.3 Monte Carlo transport code has been used to evaluate the QUADOS (Quality Assurance of Computational Tools for Dosimetry) problem P4, neutron and photon response of an albedo-type thermoluminescence personal dosemeter (TLD) located on an ISO slab phantom. Two enriched 6LiF and two 7LiF TLD chips were used and they were protected, in front or behind, with a boron-loaded dosemeter holder. Neutron response of the four chips was determined by counting 6Li(n,t)4He events using ENDF/B-VI.4 library and photon response by estimating absorbed dose (MeV g(-1)). Ten neutron energies from thermal to 20 MeV and six photon energies from 33 keV to 1.25 MeV were used to study the energy dependence. The fraction of the neutron and photon response owing to phantom backscatter has also been investigated. Detailed TRIPOLI-4.3 solutions are presented and compared with MCNP 4C calculations. PMID- 16381741 TI - Energy response of LiF and Mg2SiO4 TLDs to 10-150 keV monoenergetic photons. AB - Energy response of LiF:Mg,Ti, LiF:Mg,Cu,P and Mg2SiO4:Tb thermoluminescence dosemeters (TLDs) was measured in the range 10-150 keV for monoenergetic photons at SPring-8 of an 8-GeV synchrotron radiation facility. The photon beam was monitored by a parallel-plate free-air ionisation chamber calibrated with an uncertainty of 3%. Owing to the small dimension of the beam, a rotating holder was designed in order to irradiate TLDs uniformly. The measured responses of LiF to energy were approximately in agreement with the calculated dose absorption dependence in the soft tissue. However, two types of LiF TLDs presented the different luminescent responses to the photon energy. The response of LiF:Mg,Ti had a smooth curve, and that of LiF:Mg,Cu,P presented a local maximum at 30 keV and a local minimum at 100 keV. The Mg2SiO4:Tb response was nearly bone equivalent. Linearity of dose responses was also confirmed up to 2 Gy on each TL material. PMID- 16381742 TI - Evaluation of patient dose for mammography in Pernambuco, Brazil. AB - The aim of this study is to evaluate the incident air kerma (INAK) and the mean glandular dose (MGD) during mammography in Recife, North-east Brazil. The study was performed in three institutions: a private clinic (A), a public hospital (B) and a private hospital (C). The incident air kerma to mammography procedures were estimated using a standard breast phantom and the patient MGD, evaluated in the units B and C, was performed using the calibration of output method. This method involves the recording of the patient exposure parameters (kVp, mAs, filter) and the measurement of these parameters of the incident air kerma using a calibrated ionisation chamber. The results obtained showed that INAK for the standard breast phantom, in the institutions A, B and C, were 18.2, 9.4 and 8.5 mGy, respectively. The results of patient MGD values ranged from 0.23 to 7.46 mGy per film, and the MGD values per woman were between 8.23 and 40.6 mGy for the C institution and between 1.50 and 18.78 mGy for the B institution. PMID- 16381743 TI - On the roles of the dopants in LiF: Mg,Cu,Na,Si thermoluminescent material. AB - In this paper, some results of the study on the roles of the dopants in the LiF:Mg,Cu,Na,Si thermoluminescent (TL) material that was developed at the Korea Atomic Energy Research Institute for radiation protection are presented. Although there have been many studies to investigate the roles of the dopants in LiF:Mg,Cu,P TL material in the TL process, there are some discrepancies in the understanding of the roles of Cu and P between various researchers. In case of LiF:Mg,Cu,Na,Si TL material, there are a few studies on the roles of the dopants. Three kinds of samples in each of which one dopant is excluded, and the optimised sample, were prepared for this study. The measurements and analysis of the three dimensional TL spectra, based on the temperature, wavelength and intensity, and the glow curves for those samples are used in this study. The results show that Mg plays a role in the trapping of the charge carriers and Cu plays a role in the luminescence recombination process; however, the effect of Na and Si on the glow curve structure and the TL emission spectra is much less than that of Mg and Cu. It is considered that Na and Si each plays a role in the improvement of the luminescence efficiency. PMID- 16381744 TI - A computer program for the deconvolution of thermoluminescence glow curves. AB - A quick and efficient computer program was developed in order to resolve the peaks from the thermoluminescence (TL) glow curve. The program was designed to be easily used on any MS Windows-based computer with a graphical user interface. In this program, a new method based on the general one-trap TL equation was adopted to analyse the TL glow curve with the traditional first-order, second-order and general-order kinetics model. The method described here, general approximation, generates TL glow peaks and interpolates the relevant TL parameters from the glow data. The program was tested with simulated and experimental TL glow data and the results were found to be adequate and reliable. PMID- 16381745 TI - A time-dependent dose model for glovebox processing of fissile material. AB - A new modeling system for high-intensity neutral particle radiation fields is presented. The code PANDEMONIUM calculates external effective dose rates from neutrons and photons produced at specific locations within an industrial-size plutonium processing facility. The new version of PANDEMONIUM introduces time dependent neutronics for source multiplication coupled with transient source and detector positions. The code is designed to provide quick and acceptably accurate total effective dose estimates for scenarios and facilities for which conventional methods prove to be too impractical or costly to model. The energy range of the code has also been extended to include the effects of prompt-fission photons. PMID- 16381746 TI - Micromegas neutron beam monitor neutronics. AB - The Micromegas is a type of ionising radiation detector that consists of a gas chamber sandwiched between two parallel plate electrodes, with the gas chamber divided by a Frisch grid into drift and amplification gaps. Investigators have applied it to a number of different applications, such as charged particle, X-ray and neutron detection. A Micromegas device has been tested as a neutron beam monitor at CERN and is expected to be used for that purpose at the Spallation Neutron Source (SNS) under construction in Oak Ridge, TN. For the Micromegas to function effectively as neutron beam monitor, it should cause minimal disruption to the neutron beam in question. Specifically, it should scatter as few neutrons as possible and avoid neutron absorption when it does not contribute to generating useful information concerning the neutron beam. Here, we present the results of Monte Carlo calculations of the effect of different types of wall materials and detector gases on neutron beams and suggest methods for minimising disruption to the beam. PMID- 16381747 TI - Secondary photon fields produced in accelerator-based sources for neutron generation. AB - Neutrons can be produced with low-energy ion accelerators for many applications, such as the characterisation of neutron detectors, the irradiation of biological samples and the study of the radiation damage in electronic devices. Moreover, accelerator-based neutron sources are under development for boron neutron capture therapy (BNCT). Thin targets are used for generating monoenergetic neutrons, while thick targets are usually employed for producing more intense neutron fields. The associated photon field produced by the target nuclei may have a strong influence on the application under study. For instance, these photons can play a fundamental role in the design of an accelerator-based neutron source for BNCT. This work focuses on the measurement of the photon field associated with neutrons that are produced by 4.0-6.8 MeV protons striking both a thin 7LiF target (for generating monoenergetic neutrons) and a thick beryllium target. In both cases, very intense photon fields are generated with energy distribution extending up to several MeV. PMID- 16381748 TI - Evaluation of N50 neutron slab monitor detection efficiency with TRIPOLI-4.3 Monte Carlo code. AB - Monte Carlo code TRIPOLI-4.3 has been used to evaluate the N50 neutron slab monitor detection efficiency benchmark organised by the non-destructive assay (NDA) working group of European Safeguards Research and Development Association (ESARDA). The lattice geometry of TRIPOLI-4.3 was applied to simplify the modelling of the N50 monitor, which consists of four 3He tubes embedded in a high density polyethylene (HDPE) moderator. Using additional HDPE slabs and/or Cd sheets, 10 geometry configurations with different states of moderation were considered in this study. Two cross section libraries from ENDF/B-VI.4 and JEF2.2 and three representations of the 252Cf fission spectrum were investigated and the calculation experiment (C/E) ratios were discussed. The perturbation option was used in order to study the sensitivity to the HDPE density. The S(alpha,beta) thermal scattering in HDPE was also tested to quantify its impact on the N50 detection efficiency calculations. PMID- 16381749 TI - Analysis of shielding materials in a Compton spectrometer applied to x-ray tube quality control using Monte Carlo simulation. AB - A realistic characterisation of the primary beam is very important for the quality control of X-ray tubes. The most accurate technique to assess the actual photon spectrum is X-ray spectrometry. Some difficulties arising in the spectrum determination can be avoided using a Compton spectrometer. Simulation models are useful tools to know the effect of some operational parameters, such as collimation of primary beam, relative position of focus and detector, and the influence of shielding materials. A simulation model has been developed using the MCNP code, based on the Monte Carlo method, in order to reproduce a commercial Compton spectrometer. In this work, the model developed is applied to analyse the influence on measurements of shielding materials present in the spectrometer. PMID- 16381750 TI - Monte Carlo simulation of neutron backscattering from concrete walls in the dense plasma focus laboratory of Bologna University. AB - Between 2001 and 2003 a 3.2 kJ dense plasma focus (DPF) device has been built at the Montecuccolino Laboratory of the Department of Energy, Nuclear and Environmental Control Engineering (DIENCA) of the University of Bologna. A DPF is a pulsed device in which deuterium nuclear fusion reactions can be obtained through the pinching effects of electromagnetic fields upon a dense plasma. The empirical scale law that governs the total D-D neutron yield from a single pulse of a DPF predicts for this machine a figure of approximately 10(7) fast neutrons per shot. The aim of the present work is to evaluate the role of backscattering of neutrons from the concrete walls surrounding the Montecuccolino DPF in total neutron yield measurements. The evaluation is performed by MCNP-5 simulations that are aimed at estimating the neutron spectra at a few points of interest in the laboratory, where neutron detectors will be placed during the experimental campaigns. Spectral information from the simulations is essential because the response of detectors is influenced by neutron energy. Comparisons are made with the simple r(-2) law, which holds for a DPF in infinite vacuum. The results from the simulations will ultimately be used both in the design and optimisation of the neutron detectors and in their final calibration and placement inside the laboratory. PMID- 16381751 TI - Towards an optimum design of a P-MOS radiation detector for use in high-energy medical photon beams and neutron facilities: analysis of activation materials. AB - The behaviour of packaged and unpackaged ESAPMOS4 RadFET radiation detectors (NMRC Cork, Ireland) was investigated when used in the mixed photon and neutron environment of a medical linear accelerator operating above the nucleon separation energy and in a 14 MeV neutron field provided by a D-T generator. Within the uncertainty of the experimental set-up (4% at 95% confidence level) the unpackaged device was found to have essentially zero activation dose-burden whereas the packaged device exhibits a considerable degree of post irradiation absorbed dose due to deactivation radiation. PMID- 16381752 TI - The performance of commercial photodiodes for dosimetry in mammography. AB - This paper reports the results of an investigation carried out to determine the suitability of using a commercially available photodiode for dosimetry in mammography thereby providing a low cost dosemeter with a direct reading of the radiation dose. A mammographic X-ray generator (Siemens Mammomat 1000) with tube potential in the 23-30 kV range and a constant potential X-ray generator (Pantak) with a W/Mo anode/filter combination were used in this study. The results showed that the photodiode response is highly linear within mammographic dose ranges and that the energy dependence was <3% at tube potentials of 25-30 kV. Good agreement was observed between the incident air kerma measured with both the photodiode and the ionization chamber. These results show the viability of using the photodiode as a dosemeter system in mammography. PMID- 16381753 TI - Towards the enhancement of the photon/neutron discrimination of C6D6 detectors in the range from 1 to 10 MeV using liquid scintillator materials doped with high-Z elements. AB - The study on the behaviour of deuterated benzene detectors in the energy range from 1 to 10 MeV has been carried out. It is shown that the photon-to-neutron discrimination, attained with standard detectors, can be improved by doping the C6D6 liquid scintillator with high-Z elements. The motivation for doping the organic scintillator stems from the fact that they should increase the gamma detection efficiencies with no significant changes in the abilities to detect neutrons. The results obtained using the MCNP and EGS4 codes show that increasing the fraction of high-Z elements: (a) the energy deposited in the medium by photons increases, (b) the energy deposited by neutrons decreases and (c) the rate of detection of photon/neutron is enhanced. Owing to their low sensitivity to neutrons and the enhanced gamma detection efficiency, these detectors could be ideally used to carry out studies in mixed neutron-photon fields in the energy range considered and to assess the gamma backgrounds in noisy neutron environments. PMID- 16381754 TI - Neutron spectrometry with large volume, heavy-loaded superheated droplet detectors: a simple spin-off. AB - SIMPLE is a superheated droplet detector (SDD) experiment designed to search for the evidence of spin-dependent weakly interacting neutralino dark matter (WIMPs). SDDs, a type of emulsion detector, consist of a uniform suspension of superheated liquid droplets in a compliant material such as a polymeric or aqueous gel. We report on the first neutron spectrometry experiments with SIMPLE SDDs, a spin-off of the neutron detector calibrations performed at the Portuguese Research Reactor. SIMPLE SDDs differ from most SDDs available commercially as they have a 10 times higher loading factor, containing 10(3) times more freon than their commercial counterparts and a 100 times larger volume. We have analysed the response of SIMPLE SDDs to two quasi-monochromatic neutron beams of energies 54 and 144 keV obtained with passive filters. Results show that the characteristic peaks in the fluence distribution of both filters could be determined and their energy position obtained using a simple thermodynamic relation. PMID- 16381755 TI - The launch of MCBEND 10. AB - MCBEND 10 is the latest release of the general radiation transport Monte Carlo code from the ANSWERS Software Service of Serco Assurance. MCBEND is developed within a Nuclear Code Development (NCD) partnership between Serco Assurance and BNFL. The ANSWERS vision is 'to provide easy-to-use software that meets the current and emerging needs of the user community'. In the case of MCBEND, this vision focuses on the key areas of accuracy, understanding of uncertainties, efficiency and user-friendliness. MCBEND 10 is a major launch of the code with many new and enhanced features. New developments in MCBEND 10 include automatic splitting mesh generation, point energy adjoint for neutrons, calculation of uncertainty in the results due to material cross section uncertainties and a unified source facility. Enhanced features include improved temperature treatment, extended scoring of sensitivity to geometry perturbations, geometry improvements, extensions to formulae and improved user guide image. The user friendliness of the MCBEND code has been further enhanced by recent developments to the visualisation tools, VISAGE and VISTA-RAY. Developments have been made to the three-dimensional visualisation tool, VISTA-RAY, to simplify the detailed checking of a model, with the option to use a mouse-pointer to select regions of interest for further detail and to visually highlight incorrectly defined areas. A further development to VISTA-RAY is the inclusion of the capability to overlay a representation of a user-designated set of results from a MCBEND analysis on the model. Improvements have also been made to the graphical user interface LaunchPad for submitting and controlling calculation submission, with a common user-image across all the systems. Recent enhancements to LaunchPad include a job scheduler to simplify processing multiple tasks. A selection of the new developments in MCBEND 10, VISTA-RAY and LaunchPad will be described in this paper. PMID- 16381756 TI - Gelina neutron target optimisation. AB - A study is being performed on the properties of the Geel Electron Linear Accelerator (GELINA), a powerful white neutron source, designed for the high energy resolution time-of-flight measurements. The main aim of this study is to reduce the time spread of neutrons of the given energy without compromising the neutron yield. Both time spread and neutron intensity influence the experimental accuracy of high-resolution neutron cross section measurements, which are particularly important in the resonance region. The quantities of interest have been simulated with coupled electron-photon-neutron steady state and transient MCNP4C3 calculations. Following benchmarking of the code to the properties of the existing target, neutron yield, energy spectra, resolution functions, and neutron and heat spatial distributions have been determined for various alternative geometries and materials. At a fixed accelerator power, actinides deliver the highest neutron yield and a small target provides the best time resolution. The resulting high-power density requires a joint optimisation of the thermal hydraulics and neutronics properties. PMID- 16381757 TI - Experimental and Monte Carlo evaluation of the neutron flux of an assembly with two AmBe sources. AB - This work aimed to determine the irradiator thermal (under-cadmium) and fast (over-cadmium) neutron fluxes, of the Nuclear Experimental Laboratory of the Nuclear Engineering Center (CNEN-IPEN, Sao Paulo, Brazil), and the possibility of this irradiator use for Neutron Activation Analysis (NAA), by the absolute method. To establish the facility specifications, the neutron flux values along the irradiator axis were determined experimentally and calculated by Monte Carlo method. The irradiator presents the advantage of supplying a stable neutron flux for a long period, eliminating the need to use standard material (comparative method), so that the process becomes agile, practical and economical. PMID- 16381758 TI - Shielding for a cyclotron used for medical isotope production in China. AB - Monte Carlo and discrete ordinate calculations have been performed to determine the doses at several locations in a positron emission tomography (PET) facility in China, where the radiation source is a cyclotron that is used for the production of the isotopes necessary for PET scans. The energy-dependent neutron source term is obtained by calculations using the ALICE code, and is interpolated for input to Monte Carlo and discrete ordinate calculations. The building that houses the cyclotron has a labyrinth of walls to minimise dose to operators and to other occupants of the building. Unbiased Monte Carlo calculations did not converge after more than one week of CPU time, whereas direction biasing alone resulted in convergence in several days. A study of several biasing techniques indicated that about a factor of 3 in computational efficiency is obtained using evaluated biasing methods. The use of adjoint fluxes for biasing Monte Carlo calculations can improve computational efficiencies by one or two orders of magnitude for some problems. PMID- 16381759 TI - Dose rate calculations for a reconnaissance vehicle. AB - A Chemical Nuclear Reconnaissance System (CNRS) has been developed by the British Ministry of Defence to make chemical and radiation measurements on contaminated terrain using appropriate sensors and recording equipment installed in a land rover. A research programme is under way to develop and validate a predictive capability to calculate the build-up of contamination on the vehicle, radiation detector performance and dose rates to the occupants of the vehicle. This paper describes the geometric model of the vehicle and the methodology used for calculations of detector response. Calculated dose rates obtained using the MCBEND Monte Carlo radiation transport computer code in adjoint mode are presented. These address the transient response of the detectors as the vehicle passes through a contaminated area. Calculated dose rates were found to agree with the measured data to be within the experimental uncertainties, thus giving confidence in the shielding model of the vehicle and its application to other scenarios. PMID- 16381760 TI - Benchmarking of MCNP for calculating dose rates at an interim storage facility for nuclear waste. AB - During the operation of research facilities at Research Centre Julich, Germany, nuclear waste is stored in drums and other vessels in an interim storage building on-site, which has a concrete shielding at the side walls. Owing to the lack of a well-defined source, measured gamma spectra were unfolded to determine the photon flux on the surface of the containers. The dose rate simulation, including the effects of skyshine, using the Monte Carlo transport code MCNP is compared with the measured dosimetric data at some locations in the vicinity of the interim storage building. The MCNP data for direct radiation confirm the data calculated using a point-kernel method. However, a comparison of the modelled dose rates for direct radiation and skyshine with the measured data demonstrate the need for a more precise definition of the source. Both the measured and the modelled dose rates verified the fact that the legal limits (<1 mSv a(-1)) are met in the area outside the perimeter fence of the storage building to which members of the public have access. Using container surface data (gamma spectra) to define the source may be a useful tool for practical calculations and additionally for benchmarking of computer codes if the discussed critical aspects with respect to the source can be addressed adequately. PMID- 16381761 TI - Estimation of flux distributions with Monte Carlo functional expansion tallies. AB - Monte Carlo methods provide a powerful technique for estimating the average radiation flux in a volume (or across a surface) in cases where analytical solutions may not be possible. Unfortunately, Monte Carlo simulations typically provide only integral results and do not offer any further details about the distribution of the flux with respect to space, angle, time or energy. In the functional expansion tally (FET) a Monte Carlo simulation is used to estimate the functional expansion coefficients for flux distributions with respect to an orthogonal set of basis functions. The expansion coefficients are then used in post-processing to reconstruct a series approximation to the true distribution. Discrete event FET estimators are derived and their application in estimating radiation flux or current distributions is demonstrated. Sources of uncertainty in the FET are quantified and estimators for the statistical and truncation errors are derived. Numerical results are presented to support the theoretical development. PMID- 16381763 TI - On the high-resolution gamma-ray spectrometric measurement of 40K in natural and synthetic materials. AB - Many regulatory agencies require that all building materials and industrial waste be tested for their naturally occurring radioactive material (NORM) concentrations before they can be used or thrown away. Usually the NORM concentrations of 40K, 232Th and 238U are measured by gamma-ray spectrometry using high-purity germanium or NaI(Tl) detectors. 40K is measured through its 1460.8 keV gamma line, which is mixed with 1459.2 keV line of 228Ac from the chain of 232Th. This fact ignored till now, requires a correction in the computation of 40K concentration. Although in many cases the error is <1%, there are cases where there have been higher errors. It should be emphasised that even if the correction in 40K concentration is large, the correction of the external dose index is negligible owing to the weighing factor being higher for 232Th than for 40K (by at least a factor of 10). PMID- 16381762 TI - Radiation impact caused by activation of air from the future GSI accelerator facility fair. AB - The Gesellschaft fur Schwerionenforschung in Darmstadt is planning a new accelerator Facility for Antiproton and Ion Research (FAIR). Two future experimental areas are regarded to be the most decisive points concerning the activation of air. One is the area for the production of antiprotons. A second crucial experimental area is the so-called Super Fragment Separator. The production of radioactive isotopes in air is calculated using the residual nuclei option of the Monte Carlo program FLUKA. The results are compared with the data for the activation of air given by Sullivan and in IAEA report 283. The resulting effective dose is calculated using a program package from the German Federal Office for Radiation Protection, the Bundesamt fur Stranlenschutz. The results demonstrate that a direct emission of the total radioactivity produced into the air will probably conflict with the limits of the German Radiation Protection Ordinance. Special measures have to be planned in order to reduce the amount of radioactivity released into the air. PMID- 16381764 TI - Radiotoxicity and decay heat power of spent nuclear fuel of VVER type reactors at long-term storage. AB - Radiotoxicity and decay heat power of the spent nuclear fuel of VVER-1000 type reactors are calculated during storage time up to 300,000 y. Decay heat power of radioactive waste (radwaste) determines parameters of the heat removal system for the safe storage of spent nuclear fuel. Radiotoxicity determines the radiological hazard of radwaste after its leakage and penetration into the environment. PMID- 16381765 TI - Usefulness and limits of biological dosimetry based on cytogenetic methods. AB - Damage from occupational or accidental exposure to ionising radiation is often assessed by monitoring chromosome aberrations in peripheral blood lymphocytes, and these procedures have, in several cases, assisted physicians in the management of irradiated persons. Thereby, circulating lymphocytes, which are in the G0 stage of the cell cycle are stimulated with a mitogenic agent, usually phytohaemagglutinin, to replicate in vitro their DNA and enter cell division, and are then observed for abnormalities. Comparison with dose-response relationships obtained in vitro allows an estimate of exposure based on scoring: Unstable aberrations by the conventional, well-established analysis of metaphases for chromosome abnormalities or for micronuclei; So-called stable aberrations by the classical G-banding (Giemsa-Stain-banding) technique or by the more recently developed fluorescent in situ hybridisation (FISH) method using fluorescent labelled probes for centromeres and chromosomes. Three factors need to be considered in applying such biological dosimetry: (1) Radiation doses in the body are often inhomogeneous. A comparison of the distribution of the observed aberrations among cells with that expected from a normal poisson distribution can allow conclusions to be made with regard to the inhomogeneity of exposure by means of the so-called contaminated poisson distribution method; however, its application requires a sufficiently large number of aberrations, i.e. an exposure to a rather large dose at a high dose rate. (2) Exposure can occur at a low dose rate (e.g. from spread or lost radioactive sources) rendering a comparison with in vitro exposure hazardous. Dose-effect relationships of most aberrations that were scored, such as translocations, follow a square law. Repair intervening during exposure reduces the quadratic component with decreasing dose rate as exposure is spread over a longer period of time. No valid solution for this problem has yet been developed, although, in theory, both deterministic damage and aberrations might be repaired to a similar degree; a comparison of aberrations following a linear dose relationship might also help when the doses have been sufficiently large. (3) Investigations might have been possible only a certain time after the exposure. The relatively rapid disappearance of lymphocytes carrying unstable aberrations limits their use in retrospective dosimetry, years after exposure. Scoring stable aberrations, thought to persist in the circulating lymphocytes, might appear more appropriate in such situations. However, the examination of a representative number of cells by G-banding is extremely laborious, and the FISH method is not only expensive but has not yet been fully validated in different laboratories. In conclusion, biological dosimetry has serious limitations exactly for situations where the need for information is most urgent. It renders its most useful results when an individual has been exposed to a rather homogeneous high-level radiation over a short time interval, i.e. accidents at high-intensity radiation devices. On the other hand, it yielded less satisfactory information even when the most recent techniques were used for situations, where a low level, low dose rate exposure has occurred at some time in the past, for example for persons living in areas contaminated from the Chernobyl accident. Such negative experiences should be kept in mind in order to avoid futile and expensive investigations in the case of populations exposed from radioactivity and, notably, also from potentially clastogenic chemical agents. PMID- 16381766 TI - Use of cytogenetic indicators in radiobiology. AB - The study of ionising radiation has systematically relied on cytogenetic indicators to evaluate the biological effects and has led to theoretical approaches to explain observations associated with radiation exposure. In many of the early studies on radiobiology, the induction of chromosomal aberrations was the method of choice to evaluate dose-response relationships. But progressively, this and other cytogenetic biomarkers were used to obtain mechanistic insight on the biological effects induced by radiation. This paper attempts to give a view on the use of cytogenetic indicators in the study of various radiation-related phenomena, including radiation dosimetry, mechanisms involved in the various cellular responses to radiation, such as bystander effects, chromosomal instability and adaptive response, as well as DNA repair pathways. One future direction may involve the use of cytogenetic indicators to evaluate various molecular determinants in individuals' susceptibility to radiation, using other techniques such as fluorescence in situ hybridisation (FISH) and linking them to specific gene functions and single nucleotide polymorphisms. PMID- 16381767 TI - Assessment by cytogenetic analysis of the radioprotection properties of propolis extract. AB - Propolis obtained from honeybee hives has been used in folk medicine as an anti inflammatory, anti-carcinogenic or immunomodulatory agent. In animal studies, the radioprotector effect of propolis has been attributed to its free-radical scavenging properties. The present study was carried out to show the protective properties of propolis extract against DNA damage induced by gamma irradiation. The evaluation of the radioprotective effect of propolis has been carried out by the analysis of chromosome aberration induction after several doses of gamma rays. The results of an analysis in the presence of ethanol extract of propolis (EEP) were compared with the dose-effect calibration curve for gamma-rays by analysis of chromosome aberrations without propolis, a decrease in the radiation induced chromosome aberrations has been observed to be higher than 50% for all the doses. PMID- 16381768 TI - Radiation protection aspects of a 4 MW target. AB - The CERN Superconducting Proton Linac (SPL) is expected to provide a 2.2 GeV, 4 MW proton beam to feed facilities such as, for example, a neutrino factory or a neutrino superbeam. Material activation in such facilities is an important aspect that has to be taken into account at an early stage in designing it. In particular, the choice of the target has consequences on the induced radioactivity and dose rates in the target station and its surroundings. In the present work, the radiological aspects of a stationary target made up of tantalum pellets are compared with those of a free-surface jet of mercury. An estimation of the hadronic inelastic interactions and the production of residual nuclei in the target, the two concentric magnetic horns, the decay tunnel, the surrounding rock and a downstream dump were performed for both targets using the Monte Carlo code FLUKA. The aim was to assess the dose-equivalent rate that is to be expected during maintenance work and to evaluate the amount of residual radioactivity, which will have to be disposed of after the facility has ceased operation. The problem of after-heat in the tantalum target and the consequences of raising the proton beam energy from 2.2 to 4 GeV were also investigated. PMID- 16381769 TI - Remanent dose rates around the collimators of the LHC beam cleaning insertions. AB - The LHC will require an extremely powerful and unprecedented collimation system. As approximately 30% of the LHC beam is lost in the cleaning insertions, these will become some of the most radioactive locations around the entire LHC ring. Thus, remanent dose rates to be expected during later repair or maintenance interventions must be considered in the design phase itself. As a consequence, the beam cleaning insertions form a unique test bed for a recently developed approach to calculate remanent dose rates. A set of simulations, different in complexity, is used in order to evaluate methods for the estimation of remanent dose rates. The scope, as well as the restrictions, of the omega-factor method are shown and compared with the explicit simulation approach. The latter is then used to calculate remanent dose rates in the beam cleaning insertions. Furthermore, a detailed example for maintenance dose planning is given. PMID- 16381770 TI - Induced radioactivity in the ESRF storage ring. AB - The new French radiation protection legislation requires the definition of the zoning of accelerator facilities with respect to radioactive waste. This activation inside the ESRF 6 GeV storage ring is essentially due to photonuclear reactions. This paper describes the first results of Monte Carlo calculations that were started to prepare this zoning. The electron beam losses inside the storage ring, required to calculate saturation activities, are described. Results for the activation of the air inside the ring tunnel and of the cooling water are presented. The activation of accelerator components is illustrated with the results of the activation of the stainless steel vessels in a standard cell and in a cell with higher losses due to the presence of a vertical scraper. The amount of activation is compared with clearance levels given in the European directive 96/29/Euratom. PMID- 16381771 TI - Characterisation of radioactive waste products associated with plant decommissioning. AB - The inventory of radioactivity that must be considered in the decommissioning of a typical 1000 MWe Spanish pressurised water reactor (PWR) was investigated as part of a generic plant decommissioning study. Analyses based on DORT models (in both R-Z and R-theta geometries) were used with representative plant operating history and core power distribution data in defining the expected neutron environment in regions near the reactor core. The activation analyses were performed by multiplying the DORT scalar fluxes by energy-dependent reaction cross sections (based on ENDF/B-VI data) to generate reaction rates on a per atom basis. The results from the ORIGEN2 computer code were also used for determining the activities associated with certain nuclides where multi-group cross section data were not available. In addition to the bulk material activation of equipment and structures near the reactor, the activated corrosion-product (or 'crud') deposits on system and equipment surfaces were considered. The projected activities associated with these sources were primarily based on plant data and experience from operating PWR plants. PMID- 16381772 TI - Estimates of dose rates during the dismantling of JET. PMID- 16381773 TI - Radiation issues in a radioactive ion decay ring. AB - In a beta-beam facility, a pure beam of electron neutrinos, or their antiparticles, are produced by the decay of fully stripped radioactive ions (6He and 18Ne) circulating in a storage ring. Since the beam is not extracted from the ring, all the particles will eventually be lost somewhere in the machine and thus activate the accelerator components and the surrounding concrete and rock. In particular, as nuclei change their charge in beta-decay, a large part of the particles will be lost in the arcs of the decay ring and mainly irradiate the magnets. The density of inelastic interactions of hadrons in the magnets, concrete and rock and the track-length distribution of secondary hadrons were calculated by means of the FLUKA Monte Carlo code. These values were used to estimate the induced radioactivity in the facility, the dose rates expected in the decay ring and the consequences for the environment. PMID- 16381774 TI - Elastic image mapping for 4-D dose estimation in thoracic radiotherapy. AB - PURPOSE: Demonstrate the path integration of a four-dimensional (4-D) dose distribution onto the 3-D anatomy. MATERIALS AND METHODS: A computer-generated 4 D thoracic phantom with a lung tumour was constructed. Eight respiratory phases were generated. A radiotherapy treatment plan was applied to all the phases resulting in a 4-D dose distribution. An elastic image registration algorithm was used to find the vector displacement between all the image elements and the end expiration phase. The path-integrated tissue dose distribution and each component dose distribution were compared with the planned dose distribution. RESULTS: Numerical path integration was performed to calculate the tissue dose distribution. Loss of tumour coverage was the predominant effect observed with tumour motion in this study. The loss was asymmetric and dependent on the tumour trajectory. CONCLUSION: The elastic image registration allowed an accurate path integration through a 4-D data set to produce an accurate 3-D tissue dose estimate. PMID- 16381775 TI - Radiation transport calculations for 50 MV photon therapy beam using the Monte Carlo code GEANT4. AB - A new thin transmission target technique for fast dose delivery using narrow scanned photon beams has been developed. High-energy, 50-100 MeV, electron beams of low emittance incident on thin low-Z targets produce narrow and intense high energy bremsstrahlung beams. However, electrons transmitted through the target are bent from the therapeutic beam by a purging magnet and have to be effectively absorbed in a dedicated electron collector. The electron-photon transport through a treatment head has been studied using the Monte Carlo simulation toolkit Geant4. The Geant4 electromagnetic physics processes have been compared with experimental data of radial dose profiles. The differences between calculated and measured radial dose distributions are approximately 2-10%. Preliminary investigations of the collector design have been carried out in order to minimise secondary electron and photon contamination of the therapeutic beam. The toolkit presented here is promising for further development of narrow photon beam therapy. PMID- 16381776 TI - Measurements of secondary neutron dose from 15 MV and 18 MV IMRT. AB - Secondary neutron dose-equivalents were determined for conventional and intensity modulated radiation therapy (IMRT) prostate treatments for 15 and 18 MV X-ray beams. Conventional and IMRT treatment plans were generated to deliver 45 Gy to the prostate, seminal vessicles and external and internal iliac lymph nodes. Neutron spectra were determined by unfolding measurements from a TLD-based Bonner sphere system. Treatments using 18 MV IMRT and conventional plans result in neutron ambient dose-equivalents of 687 and 112 mSv, respectively. Delivery of the 15 MV IMRT and conventional plans results in neutron ambient dose-equivalents of 327 and 52 mSv, respectively. The data illustrate that using lower photon energies for IMRT reduces the secondary neutron dose, while still achieving comparable treatment volume coverage and sparing critical normal tissue. PMID- 16381777 TI - A CT and MRI scan to MCNP input conversion program. AB - We describe a new program to read a sequence of tomographic scans and prepare the geometry and material sections of an MCNP input file. Image processing techniques include contrast controls and mapping of grey scales to colour. The user interface provides several tools with which the user can associate a range of image intensities to an MCNP material. Materials are loaded from a library. A separate material assignment can be made to a pixel intensity or range of intensities when that intensity dominates the image boundaries; this material is assigned to all pixels with that intensity contiguous with the boundary. Material fractions are computed in a user-specified voxel grid overlaying the scans. New materials are defined by mixing the library materials using the fractions. The geometry can be written as an MCNP lattice or as individual cells. A combination algorithm can be used to join neighbouring cells with the same material. PMID- 16381778 TI - ORANGE, a new, fast dose engine for radiotherapy treatment planning. AB - A new Monte Carlo dose engine is introduced under the name of ORANGE. It is a general purpose and flexible program, because it is based on MCNP, and it is fast, because a fast dose algorithm has been added to it. In this paper it is shown that the dose results for photons and electrons are accurate within 2%, and that ORANGE can calculate accurate three-dimensional dose distributions in a 30 x 30 x 30 cm3 phantom with 60(3) voxels within a quarter of an hour on present day PCs, even before employing variance reduction techniques. PMID- 16381779 TI - Current developments at IRSN on computational tools dedicated to assessing doses for both internal and external exposure. AB - The paper presents the OEDIPE (French acronym that stands for tool for personalised internal dose assessment) and SESAME (for simulation of external source accident with medical images) computational tools, dedicated to internal and external dose assessment, respectively, and currently being developed at the Institute for Radiological Protection and Nuclear Safety. The originality of OEDIPE and SESAME, by using voxel phantoms in association with Monte Carlo codes, lies in their ability to construct personalised voxel phantoms from medical images and automatically generate the Monte Carlo input file and visualise the expected results. OEDIPE simulates in vivo measurements to improve their calibration, and calculates the dose distribution taking both internal contamination and internal radiotherapy cases into account. SESAME enables radiological overexposure doses to be reconstructed, as also victim, source and accident environment modelling. The paper presents the principles on which these tools function and an overview of specificities and results linked to their fields of application. PMID- 16381780 TI - Comparison of effective doses from various monoenergetic particles based on the stylised and the VIP-Man tomographic models. AB - This study compares the effective doses from a MIRD-type stylised model with those derived from the scaled-down version of the tomographic VIP-Man model for photon, electron, neutron and proton beams. The effective dose results from these two models show that they differ from each other within approximately 10% for common high-energy photon beams, within approximately 16% for neutrons, and within approximately 4% for high-energy proton beams. However, for low-energy protons and common electron beams, the effective doses can be different in >100%. It is concluded that the use of a single tomographic models will not improve the operational radiation protection dosimetry involving external beam exposures. PMID- 16381781 TI - Effective quality factors for neutrons based on the revised ICRP/ICRU recommendations. AB - The quality factor (Q) is intended to relate the biological effectiveness of a radiation to the absorbed dose delivered in tissue. Quality factors are defined as a function of the unrestricted linear energy transfer (L) relationship in water and are used with operational quantities. Radiation weighting factors (wR) are used in protection quantities to take into account total radiation detriment. While the International Commission on Radiological Protection (ICRP) defines the Q(L) relationship, the International Commission on Radiation Units and Measurements (ICRU) recommends the charged particle stopping power and range data. If either of these data recommendations change, the quality factors must be recomputed. The latest guidance from both organisations applicable to neutron quality factors are the ICRP Publication 60 (Q(L) relationship) and the ICRU Report 49 (stopping power and range data). In the present study, absorbed dose conversion coefficients (pGy cm2) were calculated for two operational quantities defined by the ICRU--the ambient absorbed dose and the personal absorbed dose. Dose-equivalent (pSv cm2) conversion coefficients were also computed using mean quality factors based on ICRP 60 and ICRU 49 recommendations. Effective quality factors were then calculated from the ratio of the dose-equivalent to the absorbed dose conversion coefficients for both the personal dose-equivalent and ambient dose-equivalent and compared to values reported in the literature. PMID- 16381782 TI - Analysis of QUADOS problem on TLD-ALBEDO personal dosemeter responses using discrete ordinates and Monte Carlo methods. AB - In the scope of QUADOS, a Concerted Action of the European Commission, eight calculational problems were prepared in order to evaluate the use of computational codes for dosimetry in radiation protection and medical physics, and to disseminate "good practice" throughout the radiation dosimetry community. This paper focuses on the analysis of the P4 problem on the 'TLD-albedo dosemeter: neutron and/or photon response of a four-element TL-dosemeter mounted on a standard ISO slab phantom'. Altogether 17 solutions were received from the participants, 14 of those transported neutrons and 15 photons. Most participants (16 out of 17) used Monte Carlo methods. These calculations are time-consuming, requiring several days of CPU time to perform the whole set of calculations and achieve good statistical precision. The possibility of using deterministic discrete ordinates codes as an alternative to Monte Carlo was therefore investigated and is presented here. In particular the capacity of the adjoint mode calculations is shown. PMID- 16381783 TI - Feasibility study of an active extremity dosimetry prototype. AB - In nuclear medicine departments, where radioactive sources are manipulated, the personnel can receive large radiation doses to the skin of their hands. For performing detailed characterisations and dose optimisations of these workplaces, active extremity dosemeters can be used as complementary tools to passive hand monitoring. Active extremity dosimetry is still a subject of research. In this context, IRSN has started a research and development programme. As a first step, a hospital workplace study has been performed using thermoluminescence dosemeters and has shown, in agreement with previous works, that the pads of the fingers, points that are very difficult to instrument, receive the largest doses. Numerical studies have now started, with the aim of calculating the dose equivalent gradients through the hands, in order to optimise the locations of the detectors. PMID- 16381784 TI - Shielding design for the front end of the CERN SPL. AB - CERN is designing a 2.2-GeV Superconducting Proton Linac (SPL) with a beam power of 4 MW, to be used for the production of a neutrino superbeam. The SPL front end will initially accelerate 2 x 10(14) negative hydrogen ions per second up to an energy of 120 MeV. The FLUKA Monte Carlo code was employed for shielding design. The proposed shielding is a combined iron-concrete structure, which also takes into consideration the required RF wave-guide ducts and access labyrinths to the machine. Two beam-loss scenarios were investigated: (1) constant beam loss of 1 Wm(-1) over the whole accelerator length and (2) full beam loss occurring at various locations. A comparison with results based on simplified approaches is also presented. PMID- 16381785 TI - MCNPX vs. DORT for SNS shielding design studies. AB - Radiation transport occurs through the 18 m long access way adjacent to the Spallation Neutron Source accelerator tunnel and the 2.2 m thick massive shielding door which closes the access way. A variety of typical materials for accelerator shielding, such as concrete and steel, were used for construction of the door to study radiation penetration. A comparison was carried out using both Monte Carlo (code MCNPX) and discrete ordinates (code DORT) methods. The beam losses during the accelerator operation are the sources for the radiation calculations. Analyses show that the results from the two methods are in good agreement. PMID- 16381786 TI - Radiation safety design for the J-PARC Project. AB - The High-Intensity Proton Accelerator Project, named J-PARC, is in progress, with the aim of enabling studies on the latest basic science and the advancement of nuclear technology. In the project, a high-energy proton accelerator complex with the world's highest instantaneous intensity is under construction. In order to establish a reasonable shielding design, both simplified and detailed design methods were used in the shielding design of J-PARC. This paper reviews the present status of the radiation safety design study for J-PARC. PMID- 16381787 TI - Preliminary shielding assessment for the 100 MeV proton linac (KOMAC). AB - The Proton Engineering Frontier Project is building the Korea Multipurpose Accelerator Complex facilities from 2002 to 2012, which consists of a high current 100 MeV proton linear accelerator and various beam-lines. This paper provides a preliminary estimate of the shielding required for the 20 mA proton linac and the beam-dump. For an accurate information on secondary neutron production from the guiding magnet and primary heat sink of the beam dump, proton induced 63Cu and 65Cu cross section data were evaluated and applied to shielding calculations. The required thickness of the concrete was assessed by a simple line-of-sight model for the lateral shielding of the beam-line and the full shielding of the beam dump. Monte Carlo simulations were also performed using the MCNPX code to obtain the source term and attenuation coefficients for the three dimensional lateral shielding model of the beam-line. PMID- 16381788 TI - Three-dimensional shielding calculations for the IFMIF neutron source using a coupled Monte Carlo/Deterministic computational scheme. AB - Shielding calculations for the International Fusion Materials Irradiation Facility (IFMIF) are complicated due to the geometrical complexity of the target system and the large-scale bulk shields around the source target. Three dimensional shielding calculations were performed by using a newly developed Monte Carlo/Deterministic computational scheme. The neutron-photon fluxes and dose rate distributions in the back wall of the Test Cell and the access/maintenance room are presented and compared with previous shielding calculations. The results demonstrate that this coupled scheme is an useful computational tool for three-dimensional shielding analyses of complex and large nuclear facilities. PMID- 16381789 TI - Study of the neutron beam line shield design for JSNS. AB - The JSNS, a spallation neutron source of J-PARC (JAERI-KEK Joint Project of the High Intensity Proton Accelerator) has 23 neutron beam lines. In the present study, a database was formulated for an optimum shielding design using the MCNP-X code. The calculations involved two steps. In the first step, the neutron distributions were created in the typical neutron beam line with a model that included the spallation neutron source target. The neutron currents evaluated flowed from the duct into the duct wall which was the boundary source for the bulk shield surrounding the beam line. In the second step, bulk-shield calculations were performed for the various shielding materials (iron, concrete, heavy concrete and so on) used and their composites up to thicknesses of 3 m. The results were compared with each other. Composite material shields of iron and such hydrogeneous materials as polyethylene or concrete were more effective. A typical design was prepared for a beam line within 25 m distance from a moderator, as a sample. PMID- 16381790 TI - QUADOS intercomparison: a summary of photon and charged particle problems. AB - QUADOS, a Concerted Action of the European Commission, has promoted an intercomparison aimed at evaluating the use of computational codes for dosimetry in radiation protection and medical physics. This intercomparison was open to all users of radiation transport codes. Eight problems were selected for their relevance to the radiation dosimetry community, five of which involved photon and proton transport. This paper focuses on the analysis of the photon and charged particle problems. The neutron problems were presented in a paper at the NEUDOS9 conference. PMID- 16381791 TI - Photon track evolution. AB - Given the time scale of biological, biochemical, biophysical and physical effects in a radiation exposure of living tissue, the first physical stage can be considered to be independent of time. All the physical interactions caused by the incident photons happen at the same starting time. From this point of view it would seem that the evolution of photon tracks is not a relevant topic for analysis; however, if the photon track is considered as a sequence of several interactions, there are several steps until the total degradation of the energy of the primary photon. We can characterise the photon track structure by the probability p(E,j), that is, the probability that a photon with energy E suffers j secondary interactions. The aim of this work is to analyse the photon track structure by considering j as a step of the photon track evolution towards the total degradation of the photon energy. Low energy photons (<150 keV) are considered, with water phantoms and half-extended geometry. The photon track evolution concept is presented and compared with the energy deposition along the track and also with the spatial distribution of the several steps in the photon track. PMID- 16381792 TI - Neutron-fluence-to-dose conversion coefficients in an anthropomorphic phantom. AB - A set of fluence-to-effective-dose conversion coefficients has been calculated for neutrons with energies <20 MeV using a high-resolution anthropomorphic phantom (Zubal model) and the MCNPX code. The calculation used 13 monodirectional monoenergetic neutron beams in the energy range 10(-9) to 20 MeV, under three different source irradiation configurations: anterior-posterior, posterior anterior and left lateral. Dose calculations were performed for 18 selected organs of the body, for which the International Commission on Radiological Protection and the International Commission on Radiological Units and Measurements have set tissue weighting factors for the determination of the effective dose. Another set of neutron-fluence-to-effective-dose conversion coefficients was also calculated with the proposed modification wR from ICRP Publication 92. From comparison between the dose results calculated and the data reported for the MIRD and VIPMAN models, it can be concluded that, although some discrepancies exist between the Zubal model and the two other models, there is good agreement in the left lateral irradiation geometry. PMID- 16381793 TI - Development of automatic systems for the ionising radiation metrology at the ENEA IRP secondary standard laboratory. AB - The ENEA IRP Secondary Standard Dosimetry Laboratory of Bologna (Italy) has been operated for more than 30 y as a secondary standard laboratory for X rays, gamma, beta and neutron radiation metrology. For photon radiation, the instruments for radiation protection and clinical dosimetry are calibrated in agreement with the ISO standards, using remotely controlled irradiation units: two 60Co, one 137Cs and three X-ray units (160, 320 and 420 kV, respectively). The calibration procedures, approved by the National Primary Laboratory, have been recently implemented in a software package, which controls the irradiation units and acquires the reference measurements. The system, developed at the laboratory under the LabVIEW programming language, guarantees a constant surveillance of all devices and operations, avoids procedural mistakes and automates the record keeping and reporting activities. This results in an increase in the reliability of the calibration service. PMID- 16381794 TI - An approach to 3D dose mapping using Gafchromic film. AB - The tissue equivalent composition of the Gafchromic films makes them particularly suitable for the mapping of 2D and 3D treatment fields. This paper presents the results obtained using MD-55 film for the verification of real radiotherapy treatments through proton beam irradiation of suitable phantoms at the Clatterbridge Centre for Oncology (CCO) in Bebington (UK). After exposure, the variation in optical density of the films was measured using a CCD100 Microdensitometer (source at 665 nm). Holes of calibrated diameter, made during the assembly phase of the phantom, are identified by the MIRA software, used for data analysis, and allow the rendering of the films. The surface dose distributions were obtained from the variation in optical density of each of the films making up the phantom. Their elaboration to duplicate their position within the phantom, performed by 3D-doctor software, allows the volumetric reconstruction of the dose distribution. PMID- 16381795 TI - Measurement and analysis of induced activities in concrete irradiated using high energy neutrons at KENS Neutron Spallation Source Facility. AB - Precise estimation of induced activities in concrete shields for high-energy accelerator facilities is one of the most important issues that need to be solved, not only for the reduction of exposure for workers, but also for the reduction of radioactive wastes. Irradiation experiments have been performed by using the 500 MeV Neutron Spallation Source Facility in KEK. The large concrete assembly was placed in the direction of 0 degrees to the beamline. Two kinds of samples were placed at several positions in the assembly. The irradiation period was about 1 week and induced activities in the samples were measured until approximately 1.5 y after irradiation. From the comparison between the experiment and the available Monte Carlo calculation code system, good agreement was obtained for 24Na, 47Sc, 47Ca and 54Mn within a factor 2; however, large discrepancies were observed for some other nuclides. PMID- 16381796 TI - Spatial distributions of residuals produced inside a spallation target. AB - The spallation target model of an accelerator driven system (ADS), consisting of six 5 cm thick and 16 cm in diameter Pb segments, was constructed. Three sets of 17 Bi samples (1/2 inch in diameter and 1 mm thick) were placed in 3 Pb disc shaped holders inside the target at 5, 10 and 15 cm from its front. After irradiation with 660 MeV proton beam gamma-spectra of radioisotopes produced in Bi were collected several times for each sample with the use of HPGe detectors in order to identify the radioisotopes and to determine their absolute activities. Their spatial distributions were then compared with respective values obtained in the calculations made with the use of FLUKA and/or MCNPX code. A fair agreement with the experiment has been observed. PMID- 16381797 TI - Depolarization-induced slow Ca2+ transients stimulate transcription of IL-6 gene in skeletal muscle cells. AB - Contracting skeletal muscle produces and releases interleukin-6 (IL-6) in high amounts. Nevertheless, the mechanisms underlying IL-6 expression are not understood. Because inositol-1,4,5-trisphosphate (IP(3))-mediated slow Ca(2+) signals evoked by depolarization of skeletal myotubes appears to play a role in the regulation of gene expression, we examined its involvement on IL-6 transcription. With the use of semiquantitative RT-PCR, we have shown that K(+) depolarization of myotubes induces a transient increase in IL-6 mRNA level, which peaks at 3-4 h and is independent of extracellular Ca(2+). Inhibitors of IP(3) dependent Ca(2+) signals, like 2-aminoethoxydiphenyl borate (2-APB) and U-73122, decreased activation of IL-6 gene expression as did Ca(2+) signals inhibitor BAPTA-AM, whereas ryanodine, a fast Ca(2+) transient inhibitor, had no effect on IL-6 induction. Depolarization of myotubes transiently transfected with a reporter gene construct, containing 651 bp of IL-6 promoter, induced a twofold increase in promoter activity, which was abolished by either 2-APB or U-73122 and remained unaffected after ryanodine treatment. Site-directed mutagenesis of parental construct allowed us to identify activator protein-1 and NF-kappaB sequences as regulatory elements involved in IL-6 upregulation. Our results provide evidence for involvement of IP(3)-mediated Ca(2+) signals on IL-6 transcription in skeletal muscle cells. PMID- 16381798 TI - Spatial and temporal mapping of pacemaker activity in interstitial cells of Cajal in mouse ileum in situ. AB - Spontaneous electrical pacemaker activity occurs in tunica muscularis of the gastrointestinal tract and drives phasic contractions. Interstitial cells of Cajal (ICC) are the pacemaker cells that generate and propagate electrical slow waves. We used Ca(2+) imaging to visualize spontaneous rhythmicity in ICC in the myenteric region (ICC-MY) of the murine small intestine. ICC-MY, verified by colabeling with Kit antibody, displayed regular Ca(2+) transients that occurred after electrical slow waves. ICC-MY formed networks, and Ca(2+) transient wave fronts propagated through the ICC-MY networks at approximately 2 mm/s and activated attached longitudinal muscle fibers. Nicardipine blocked Ca(2+) transients in LM but had no visible effect on the transients in ICC-MY. beta Glycyrrhetinic acid reduced the coherence of propagation, causing single cells to pace independently. Thus, virtually all ICC-MYs are spontaneously active, but normal activity is organized into propagating wave fronts. Inhibitors of dihydropyridine-resistant Ca(2+) entry (Ni(2+) and mibefradil) and elevated external K(+) reduced the coherence and velocity of propagation, eventually blocking all activity. The mitochondrial uncouplers, FCCP, and antimycin and the inositol 1,4,5-trisphosphate receptor-inhibitory drug, 2-aminoethoxydiphenyl borate, abolished rhythmic Ca(2+) transients in ICC-MY. These data show that global Ca(2+) transients in ICC-MYs are a reporter of electrical slow waves in gastrointestinal muscles. Imaging of ICC networks provides a unique multicellular view of pacemaker activity. The activity of ICC-MY is driven by intracellular Ca(2+) handling mechanisms and entrained by voltage-dependent Ca(2+) entry and coupling of cells via gap junctions. PMID- 16381799 TI - Glucose-induced mixed [Ca2+]c oscillations in mouse beta-cells are controlled by the membrane potential and the SERCA3 Ca2+-ATPase of the endoplasmic reticulum. AB - Stimulatory concentrations of glucose induce two patterns of cytosolic Ca2+ concentration ([Ca2+]c) oscillations in mouse islets: simple or mixed. In the mixed pattern, rapid oscillations are superimposed on slow ones. In the present study, we examined the role of the membrane potential in the mixed pattern and the impact of this pattern on insulin release. Simultaneous measurement of [Ca2+]c and insulin release from single islets revealed that mixed [Ca2+]c oscillations triggered synchronous oscillations of insulin secretion. Simultaneous recordings of membrane potential in a single beta-cell within an islet and of [Ca2+]c in the whole islet demonstrated that the mixed pattern resulted from compound bursting (i.e., clusters of membrane potential oscillations separated by prolonged silent intervals) that was synchronized in most beta-cells of the islet. Each slow [Ca2+]c increase during mixed oscillations was due to a progressive summation of rapid oscillations. Digital image analysis confirmed the good synchrony between subregions of an islet. By contrast, islets from sarco(endo)plasmic reticulum Ca2+-ATPase isoform 3 (SERCA3) knockout mice did not display typical mixed [Ca2+]c oscillations in response to glucose. This results from a lack of progressive summation of rapid oscillations and from altered spontaneous electrical activity, i.e., lack of compound bursting, and membrane potential oscillations characterized by lower-frequency but larger-depolarization phases than observed in SERCA3+/+ beta-cells. We conclude that glucose-induced mixed [Ca2+]c oscillations result from compound bursting in all beta-cells of the islet. Disruption of SERCA3 abolishes mixed [Ca2+]c oscillations and augments beta-cell depolarization. This latter observation indicates that the endoplasmic reticulum participates in the control of the beta-cell membrane potential during glucose stimulation. PMID- 16381800 TI - Sterol regulatory element-binding protein 1 is negatively modulated by PKA phosphorylation. AB - Sterol regulatory element-binding protein (SREBP)-1a and -1c are key transcription factors that regulate lipid biosynthesis in cells. We identified that Ser338 located at the NH2 terminus of SREBP-1a is a PKA phosphorylation site in vitro and in HepG2 cells. PKA phosphorylation of this site attenuated DNA occupancy, as revealed by chromatin immunoprecipitation assay, and the ensuing transactivation. In contrast, replacing Ser with Ala [SREBP-1a(N)-S338A] increased transactivation. Although it forms heterodimers with the wild-type SREBP-1a(N) or S338A but not a homodimer with itself, SREBP-1a(N)-S338D (replacing Ser with Asp) decreased DNA binding. Ser314 of SREBP-1c, the counterpart of SREBP-1a Ser338, was also phosphorylated by PKA. Accordingly, the adenovirus-mediated expression of SREBP-1c(N)-S314D in HepG2 cells retarded lipogenesis. Our results indicate that the cAMP-PKA pathway, by phosphorylating SREBP-1, may modulate lipid metabolism in liver cell lines. PMID- 16381801 TI - Compensatory contribution of Cav2.3 channels to acetylcholine release at the neuromuscular junction of tottering mice. AB - Tottering (Tg) mice carry the mutation P601L in their Cacna1a encoded Cav2.1 channels. Transmitter release at the wild-type neuromuscular junction (NMJ) is almost exclusively mediated by Cav2.1 channels, and we used this model synapse to study synaptic consequences of the Tg mutation. With electrophysiology, and using subtype-specific Cav2 channel-blocking toxins, we assessed a possible compensatory contribution of non-Cav2.1 channels to evoked acetylcholine (ACh) release at Tg NMJs. Release was reduced by approximately 75% by the Cav2.1 channel blocker omega-agatoxin-IVA, which was less than the approximately 95% reduction observed in wild-type. Release at Tg NMJs, but not at wild-type synapses, was reduced by approximately 15% by SNX-482, a Cav2.3 channel blocker. No Cav2.2 channel involvement was found. Probably, there is a small reduction in functional presynaptic Cav2.1 channels at Tg NMJs, which is compensated for by Cav2.3 channels. The remaining Cav2.1 channels are likely to convey enlarged Ca2+ flux, because evoked ACh release at Tg NMJs, at low extracellular Ca2+ concentration, was approximately sixfold higher than at wild-type NMJs. This is the first report of compensatory expression of non-Cav2.1 channels at NMJs of mice with a single amino acid change in Cav2.1. PMID- 16381802 TI - Altered inhibition in lateral amygdala networks in a rat model of temporal lobe epilepsy. AB - Clinical and experimental evidence indicates that the amygdala is involved in limbic seizures observed in patients with temporal lobe epilepsy. Here, we used simultaneous field and intracellular recordings from horizontal brain slices obtained from pilocarpine-treated rats and age-matched nonepileptic controls (NECs) to shed light on the electrophysiological changes that occur within the lateral nucleus (LA) of the amygdala. No significant differences in LA neuronal intrinsic properties were observed between pilocarpine-treated and NEC tissue. However, spontaneous field activity could be recorded in the LA of 21% of pilocarpine-treated slices but never from NECs. At the intracellular level, this network activity was characterized by robust neuronal firing and was abolished by glutamatergic antagonists. In addition, we could identify in all pilocarpine treated LA neurons: 1) large amplitude depolarizing postsynaptic potentials (PSPs) and 2) a lower incidence of spontaneous hyperpolarizing PSPs as compared with NECs. Single-shock stimulation of LA networks in the presence of glutamatergic antagonists revealed a biphasic inhibitory PSP (IPSP) in both NECs and pilocarpine-treated tissue. The reversal potential of the early GABA(A) receptor-mediated component, but not of the late GABA(B) receptor-mediated component, was significantly more depolarized in pilocarpine-treated slices. Furthermore, the peak conductance of both fast and late IPSP components had significantly lower values in pilocarpine-treated LA cells. Finally, paired-pulse stimulation protocols in the presence of glutamatergic antagonists revealed a less pronounced depression of the second IPSP in pilocarpine-treated slices compared with NECs. Altogether, these findings suggest that alterations in both pre- and postsynaptic inhibitory mechanisms contribute to synaptic hyperexcitability of LA networks in epileptic rats. PMID- 16381803 TI - Relief of synaptic depression produces long-term enhancement in thalamocortical networks. AB - Thalamocortical synapses may be able to undergo activity-dependent long-term changes in efficacy, such as long-term potentiation. Indeed, studies conducted in vivo have found that theta-burst stimulation (TBS) of the thalamus induces a long term enhancement (LTE) of field potential responses evoked in the neocortex of adult rodents. Because the thalamus and neocortex form a complex interconnected network that is highly active in vivo, it is possible that a change in thalamic excitability would be reflected in the neocortex. To test this possibility, we recorded from barrel neocortex and applied TBS to the thalamic radiation while the somatosensory thalamus was inactivated with muscimol. Thalamocortical LTE was absent when the thalamus was inactivated, suggesting that changes in thalamic excitability are involved. Single-unit recordings from thalamocortical cells revealed that TBS causes a significant reduction in the spontaneous firing rate of thalamocortical cells. Reducing the spontaneous firing of thalamocortical cells directly enhances the efficacy of the thalamocortical pathway because it relieves the tonic depression of the thalamocortical connection caused by thalamocortical activity. Because these changes in thalamic excitability are triggered by corticothalamic activity, this may be a useful top-down mechanism to regulate afferent sensory input to the neocortex during behavior as a function of experience. PMID- 16381804 TI - Circuit properties generating gamma oscillations in a network model of the olfactory bulb. AB - The study of the neural basis of olfaction is important both for understanding the sense of smell and for understanding the mechanisms of neural computation. In the olfactory bulb (OB), the spatial patterning of both sensory inputs and synaptic interactions is crucial for processing odor information, although this patterning alone is not sufficient. Recent studies have suggested that representations of odor may already be distributed and dynamic in the first olfactory relay. The growing evidence demonstrating a functional role for the temporal structure of bulbar neuronal activity supports this assumption. However, the detailed mechanisms underlying this temporal structure have never been thoroughly studied. Our study focused on gamma (40-100 Hz) network oscillations in the mammalian OB, which is a form of temporal patterning in bulbar activity elicited by olfactory stimuli. We used computational modeling combined with electrophysiological recordings to investigate the basic synaptic organization necessary and sufficient to generate sustained gamma rhythms. We found that features of gamma oscillations obtained in vitro were identical to those of a model based on lateral inhibition as the coupling modality (i.e., low irregular firing rate and high oscillation stability). In contrast, they differed substantially from those of a model based on lateral excitatory coupling (i.e., high regular firing rate and instable oscillations). Therefore we could precisely tune the oscillation frequency by changing the kinetics of inhibitory events supporting the lateral inhibition. Moreover, gradually decreasing GABAergic synaptic transmission decreased the degree of relay neuron synchronization in response to sensory inputs, both theoretically and experimentally. Thus we have shown that lateral inhibition provides a mechanism by which the dynamic processing of odor information might be finely tuned within the OB circuit. PMID- 16381805 TI - Altered cortical glutamate receptor function in the R6/2 model of Huntington's disease. AB - Alterations in pyramidal neurons from the sensorimotor cortex may be responsible for some of the cognitive and motor symptoms of Huntington's disease (HD). The present experiments used R6/2 transgenic mice that express exon 1 of the human HD gene with an expanded number of CAG repeats. We characterized alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) currents and their modulation by cyclothiazide (CTZ) as well as N-methyl-D-aspartate (NMDA) currents and their Mg2+ sensitivity in acutely dissociated cortical pyramidal neurons in R6/2 transgenic and wild-type (WT) mice at 21 days (before overt symptoms), 40 days (when symptoms begin), and 80 days (fully symptomatic). AMPA currents, alone or in the presence of CTZ, were smaller in 21- and 40-day-old R6/2 groups compared with WT mice. In R6/2 mice, more neurons displayed desensitizing AMPA currents in the presence of CTZ, indicating increased expression of "flop" splice variants, whereas the majority of WT cells expressed the "flip" variants of AMPA receptor subunits. NMDA peak currents also were smaller in R6/2 pyramidal neurons at 21 days. At 40 days, NMDA currents were similar in WT and R6/2 mice but Mg2+ sensitivity was greater in R6/2 mice, resulting in smaller NMDA currents in the presence of Mg2+. Differences in AMPA and NMDA currents between WT and R6/2 cells were no longer detected at 80 days. Our findings indicate that currents induced by glutamate receptor agonists are decreased in isolated cortical pyramidal neurons from R6/2 mice and that this decrease occurs early. Altered glutamate receptor function could contribute to changes in cortical output and may underlie some of the cognitive and motor impairments in this animal model of HD. PMID- 16381806 TI - Effects of end-goal on hand shaping. AB - The aim of the present study was to determine whether hand shaping was affected by planning of an action subsequent to object contact. Ten subjects (5 females and 5 males, ages 19-33) were requested to reach toward and grasp a convex object between the thumb and the four fingers of the right hand and to perform one of the following actions: 1) lift up the object; 2) insert the object into a niche of a similar shape and size as the object, or 3) insert the object into a rectangular niche much larger than the object. Flexion/extension at the metacarpal-phalangeal and proximal interphalangeal joints of all digits were measured using resistive sensors embedded in a glove. Although all experimental conditions required grasping the same object, we found different covariation patterns among finger joint angles across conditions. Gradual preshaping of the hand occurred only when planning object lift or when the end-goal required object placement into the tight niche. In contrast, for the larger niche, gradual preshaping was not evident for the ring and the little finger. Further, reaching movements were faster for movements ending with the larger niche than for the other movement conditions. The present results suggest that hand shaping takes into account end-goal in addition to object geometry. We discuss these findings in the context of forward internal models that allow the prediction of the sensorimotor consequences of motor commands in advance to their execution. PMID- 16381807 TI - Do cortical neurons process luminance or contrast to encode surface properties? AB - On the one hand, contrast signals provide information about surface properties, such as reflectance, and patchy illumination conditions, such as shadows. On the other hand, processing of luminance signals may provide information about global light levels, such as the difference between sunny and cloudy days. We devised models of contrast and luminance processing, using principles of logarithmic signal coding and half-wave rectification. We fit each model to individual response profiles obtained from 67 surface-responsive macaque V1 neurons in a center-surround paradigm similar to those used in human psychophysical studies. The most general forms of the luminance and contrast models explained, on average, 73 and 87% of the response variance over the sample population, respectively. We used a statistical technique, known as Akaike's information criterion, to quantify goodness of fit relative to number of model parameters, giving the relative probability of each model being correct. Luminance models, having fewer parameters than contrast models, performed substantially better in the vast majority of neurons, whereas contrast models performed similarly well in only a small minority of neurons. These results suggest that the processing of local and mean scene luminance predominates over contrast integration in surface responsive neurons of the primary visual cortex. The sluggish dynamics of luminance-related cortical activity may provide a neural basis for the recent psychophysical demonstration that luminance information dominates brightness perception at low temporal frequencies. PMID- 16381808 TI - Rostral ganglia are required for induction but not expression of crayfish escape reflex habituation: role of higher centers in reprogramming low-level circuits. AB - It is widely assumed that learning results from alterations in the strength of synapses within the neural pathways that mediate a learned behavioral response and that these alterations are directly caused by training-induced activity of neurons connected by the changing synapses. Initial evidence for this view came from studies of habituation of defensive reflexes in several invertebrate species. However, more recent studies of habituation of the escape reflex in one of these species, the crayfish, have shown that habituation is substantially caused by tonic inhibitory input from cephalic ganglia; this descending inhibition suppresses the activity of neurons within the escape circuit, which reside in caudal ganglia. Such control by descending inhibition indicates that animals with encephalized nervous systems do not entirely abdicate to low-level circuitry the important decision of whether to habituate to stimuli that might warn of danger. Higher centers in fact play a major role in controlling the habituation of this potentially life-saving protective response. Another way for higher centers to control lower ones would be to induce alteration of the lower center's intrinsic properties. Here, we show that, whereas descending input from higher ganglia is needed to induce habituation, once established, habituation persists even after rostral ganglia are disconnected. This provides evidence that lower-level neural circuits can be reprogrammed through transient interaction with higher ganglia to decrease their intrinsic tendency to produce escape. PMID- 16381809 TI - Inflammatory mediators enhance the excitability of chronically compressed dorsal root ganglion neurons. AB - A laterally herniated disk, spinal stenosis, and various degenerative or traumatic diseases of the spine can sometimes lead to a chronic compression and inflammation of the dorsal root ganglion and chronic abnormal sensations including pain. After a chronic compression of the dorsal root ganglion (CCD) in rats, the somata in the dorsal root ganglion (DRG) become hyperexcitable, and some exhibit ectopic, spontaneous activity (SA). Inflammatory mediators have a potential role in modulating the excitability of DRG neurons and therefore may contribute to the neuronal hyperexcitability after CCD. In this study, an inflammatory soup (IS) consisting of bradykinin, serotonin, prostaglandin E2, and histamine (each 10(-6) M) was applied topically to the DRG. The responses of DRG neurons were electrophysiologically recorded extracellularly from teased dorsal root fibers or intracellularly from the somata in the intact DRG or from dissociated neurons within 30 h of culture. In all three preparations, IS remarkably increased the discharge rates of SA CCD neurons and evoked discharges in more silent-CCD than control neurons. IS slightly depolarized the resting membrane potential and decreased the current and voltage thresholds of action potential in both intact and dissociated neurons, although the magnitude of depolarization or decrease in action potential threshold was not significantly different between CCD and control. IS-evoked responses were found in a proportion of neurons in each size category including those with and without nociceptive properties. Inflammatory mediators, by increasing the excitability of DRG somata, may contribute to CCD-induced neuronal hyperexcitability and to hyperalgesia and tactile allodynia. PMID- 16381810 TI - Glycine-gated chloride channels depress synaptic transmission in rat hippocampus. AB - An inhibitory role for strychnine-sensitive glycine-gated chloride channels (GlyRs) in mature hippocampus is beginning to be appreciated. We have reported previously that CA1 pyramidal cells and GABAergic interneurons recorded in 3- to 4-wk-old rat hippocampal slices express functional GlyRs, dispelling previous misconceptions that GlyR expression ceases in early development. However, the effect of GlyR activation on cell excitability and synaptic circuits in hippocampus has not been fully explored. Using whole cell current-clamp recordings, we show that activation of strychnine-sensitive GlyRs through exogenous glycine application causes a significant decrease in input resistance and prevents somatically generated action potentials in both CA1 pyramidal cells and interneurons. Furthermore, GlyR activation depresses the synaptic network by reducing suprathreshold excitatory postsynaptic potentials (EPSPs) to subthreshold events in both cell types. Blockade of postsynaptic GlyRs with the chloride channel blocker 4, 4'-diisothiocyanatostilbene-2-2'-disulfonic acid (DIDS) or altering the chloride ion driving force in recorded cells attenuates the synaptic depression, strongly indicating that a postsynaptic mechanism is responsible. Increasing the local glycine concentration by blocking reuptake causes a strychnine-sensitive synaptic depression in interneuron recordings, suggesting that alterations in extracellular glycine will impact excitability in hippocampal circuits. Finally, using immunohistochemical methods, we show that glycine and the glycine transporter GlyT2 are co-localized selectively in GABAergic interneurons, indicating that interneurons contain both inhibitory neurotransmitters. Thus we report a novel mechanism whereby activation of postsynaptic GlyRs can function to depress activity in the synaptic network in hippocampus. Moreover, the co-localization of glycine and GABA in hippocampal interneurons, similar to spinal cord, brain stem, and cerebellum, suggests that this property is likely to be a general characteristic of inhibitory interneurons throughout the CNS. PMID- 16381811 TI - Cdc42 and actin control polarized expression of TI-VAMP vesicles to neuronal growth cones and their fusion with the plasma membrane. AB - Tetanus neurotoxin-insensitive vesicle-associated membrane protein (TI-VAMP) mediated fusion of intracellular vesicles with the plasma membrane is crucial for neurite outgrowth, a pathway not requiring synaptobrevin-dependent exocytosis. Yet, it is not known how the TI-VAMP membrane trafficking pathway is regulated or how it is coordinated with cytoskeletal dynamics within the growth cone that guide neurite outgrowth. Here, we demonstrate that TI-VAMP, but not synaptobrevin 2, concentrates in the peripheral, F-actin-rich region of the growth cones of hippocampal neurons in primary culture. Its accumulation correlates with and depends upon the presence of F-actin. Moreover, acute stimulation of actin remodeling by homophilic activation of the adhesion molecule L1 induces a site directed, actin-dependent recruitment of the TI-VAMP compartment. Expression of a dominant-positive mutant of Cdc42, a key regulator of cell polarity, stimulates formation of F-actin- and TI-VAMP-rich filopodia outside the growth cone. Furthermore, we report that Cdc42 activates exocytosis of pHLuorin tagged TI-VAMP in an actin-dependent manner. Collectively, our data suggest that Cdc42 and regulated assembly of the F-actin network control the accumulation and exocytosis of TI-VAMP-containing membrane vesicles in growth cones to coordinate membrane trafficking and actin remodeling during neurite outgrowth. PMID- 16381812 TI - Sphingoid base is required for translation initiation during heat stress in Saccharomyces cerevisiae. AB - Sphingolipids are required for many cellular functions including response to heat shock. We analyzed the yeast lcb1-100 mutant, which is conditionally impaired in the first step of sphingolipid biosynthesis and shows a strong decrease in heat shock protein synthesis and viability. Transcription and nuclear export of heat shock protein mRNAs is not affected. However, lcb1-100 cells exhibited a strong decrease in protein synthesis caused by a defect in translation initiation under heat stress conditions. The essential lipid is sphingoid base, not ceramide or sphingoid base phosphates. Deletion of the eIF4E-binding protein Eap1p in lcb-100 cells restored translation of heat shock proteins and increased viability. The translation defect during heat stress in lcb1-100 was due at least partially to a reduced function of the sphingoid base-activated PKH1/2 protein kinases. In addition, depletion of the translation initiation factor eIF4G was observed in lcb1-100 cells and ubiquitin overexpression allowed partial recovery of translation after heat stress. Taken together, we have shown a requirement for sphingoid bases during the recovery from heat shock and suggest that this reflects a direct lipid-dependent signal to the cap-dependent translation initiation apparatus. PMID- 16381813 TI - Establishment of polarity during organization of the acentrosomal plant cortical microtubule array. AB - The plant cortical microtubule array is a unique acentrosomal array that is essential for plant morphogenesis. To understand how this array is organized, we exploited the microtubule (+)-end tracking activity of two Arabidopsis EB1 proteins in combination with FRAP (fluorescence recovery after photobleaching) experiments of GFP-tubulin to examine the relationship between cortical microtubule array organization and polarity. Significantly, our observations show that the majority of cortical microtubules in ordered arrays, within a particular cell, face the same direction in both Arabidopsis plants and cultured tobacco cells. We determined that this polar microtubule coalignment is at least partially due to a selective stabilization of microtubules, and not due to a change in microtubule polymerization rates. Finally, we show that polar microtubule coalignment occurs in conjunction with parallel grouping of cortical microtubules and that cortical array polarity is progressively enhanced during array organization. These observations reveal a novel aspect of plant cortical microtubule array organization and suggest that selective stabilization of dynamic cortical microtubules plays a predominant role in the self-organization of cortical arrays. PMID- 16381814 TI - Phosphorylation of the chromosomal passenger protein Bir1 is required for localization of Ndc10 to the spindle during anaphase and full spindle elongation. AB - The Saccharomyces cerevisiae inhibitor of apoptosis (IAP) repeat protein Bir1 localizes as a chromosomal passenger. A deletion analysis of Bir1 identified two regions important for function. The C-terminal region is essential for growth, binds Sli15, and is necessary and sufficient for the localization of Bir1 as a chromosomal passenger. The middle region is not essential but is required to localize the inner kinetochore protein Ndc10 to the spindle during anaphase and to the midzone at telophase. In contrast, precise deletion of the highly conserved IAP repeats conferred no phenotype and did not alter the cell cycle delay caused by loss of cohesin. Bir1 is phosphorylated in a cell cycle-dependent manner. Mutation of all nine CDK consensus sites in the middle region of Bir1 significantly decreased the level of phosphorylation and blocked localization of Ndc10 to the spindle at anaphase. Moreover, immunoprecipitation of Ndc10 with Bir1 was dependent on phosphorylation. The loss of Ndc10 from the anaphase spindle prevented elongation of the spindle beyond 7 microm. We conclude that phosphorylation of the middle region of Bir1 is required to bring Ndc10 to the spindle at anaphase, which is required for full spindle elongation. PMID- 16381815 TI - Rck2 is required for reprogramming of ribosomes during oxidative stress. AB - Rck2 is a mitogen-activated protein kinase-activated protein kinase in yeast implicated in translational regulation. rck2Delta mutants are mildly sensitive to oxidative stress, a condition that causes dissociation of actively translating ribosomes (polysomes). In rck2Delta cells, polysomes are lost to an even higher degree than in the wild-type upon stress. Cells overexpressing the catalytically inactive rck2-kd allele are highly sensitive to oxidative stress. In such cells, dissociation of polysomes upon stress was instead greatly delayed. The protein synthesis rate decreased to a similar degree as in wild-type cells, however, indicating that in rck2-kd cells, the polysome complexes were inactive. Array analyses of total and polysome-associated mRNAs revealed major deregulation of the translational machinery in rck2 mutant cells. This involves transcripts for cytosolic ribosomal proteins and for processing and assembly of ribosomes. In rck2Delta cells, weakly transcribed mRNAs associate more avidly with polysomes than in wild-type cells, whereas the opposite holds true for rck2-kd cells. This is consistent with perturbed regulation of translation elongation, which is predicted to alter the ratio between mRNAs with and without strong entry sites at ribosomes. We infer that imbalances in the translational apparatus are a major reason for the inability of these cells to respond to stress. PMID- 16381816 TI - The association of Shiga-like toxin with detergent-resistant membranes is modulated by glucosylceramide and is an essential requirement in the endoplasmic reticulum for a cytotoxic effect. AB - Receptor-mediated internalization to the endoplasmic reticulum (ER) and subsequent retro-translocation to the cytosol are essential sequential processes required for the productive intoxication of susceptible mammalian cells by Shiga like toxin-1 (SLTx). Recently, it has been proposed that the observed association of certain ER-directed toxins and viruses with detergent-resistant membranes (DRM) may provide a general mechanism for their retrograde transport to endoplasmic reticulum (ER). Here, we show that DRM recruitment of SLTx bound to its globotriosylceramide (Gb(3)) receptor is mediated by the availability of other glycosphingolipids. Reduction in glucosylceramide (GlcCer) levels led to complete protection against SLTx and a reduced cell surface association of bound toxin with DRM. This reduction still allowed efficient binding and transport of the toxin to the ER. However, toxin sequestration within DRM of the ER was abolished under reduced GlcCer conditions, suggesting that an association of toxin with lipid microdomains or rafts in the ER (where these are defined by detergent insolubility) is essential for a later step leading to or involving retro-translocation of SLTx across the ER membrane. In support of this, we show that a number of ER residents, proteins intimately involved in the process of ER dislocation of misfolded proteins, are present in DRM. PMID- 16381817 TI - The E7 proteins of low- and high-risk human papillomaviruses share the ability to target the pRB family member p130 for degradation. AB - High-risk human papillomaviruses (HPVs) (e.g., HPV-16) cause anogenital and head and neck cancers, and low-risk HPVs (e.g., HPV-6) cause benign hyperproliferative disease. The E7 protein of HPV-16 binds all retinoblastoma tumor suppressor protein (pRB) family members with higher affinity than HPV-6E7. The HPV-16 E7 protein has been reported to target pRB family members for degradation and to immortalize cells. Here we tested the hypothesis that the low-risk E7 protein has an intrinsic ability to decrease expression of pRB family members. First, we introduced a high-affinity pRB-binding site into HPV-6 E7 (6E7G22D) and showed that, in human foreskin keratinocytes, HPV-6 E7G22D decreased the level of pRB protein but not pRB mRNA. Second, we analyzed the ability of wild-type HPV-6 E7 to destabilize the other pRB family members, p107 and p130. HPV-6 E7, like HPV-16 E7, decreased the level of p130 protein. This decrease was inhibited by MG132, a proteasome inhibitor. Binding of HPV-6 E7 to p130 was necessary but not sufficient to decrease the level of p130. Furthermore, the destabilization of p130 correlated with a decrease in the expression of involucrin, a differentiation marker. We suggest that the shared activity of HPV-16 E7 and HPV 6 E7 to destabilize p130 and decrease or delay differentiation may be related to the role of E7 in the HPV life cycle. The added ability of HPV-16 E7 to regulate pRB and p107 may be related to oncogenic activity. PMID- 16381818 TI - Transcriptional response of steady-state yeast cultures to transient perturbations in carbon source. AB - To understand the dynamics of transcriptional response to changing environments, well defined, easily controlled, and short-term perturbation experiments were undertaken. We subjected steady-state cultures of Saccharomyces cerevisiae in chemostats growing on limiting galactose to two different size pulses of glucose, well known to be a preferred carbon source. Although these pulses were not large enough to change growth rates or cell size, approximately 25% of the genes changed their expression at least 2-fold. Using DNA microarrays to estimate mRNA abundance, we found a number of distinguishable patterns of transcriptional response among the many genes whose expression changed. Many of these genes were already known to be regulated by particular transcription factors; we estimated five potentially relevant transcription factor activities from the observed changes in gene expression (i.e., Mig1p, Gal4p, Cat8p, Rgt1p, Adr1p, and Rcs1p). With these estimates, for two regulatory circuits involving interaction among multiple regulators we could generate dynamical models that quantitatively account for the observed transcriptional responses to the transient perturbations. PMID- 16381819 TI - Recombination, rearrangement, reshuffling, and divergence in a centromeric region of rice. AB - Centromeres have many unusual biological properties, including kinetochore attachment and severe repression of local meiotic recombination. These properties are partly an outcome, partly a cause, of unusual DNA structure in the centromeric region. Although several plant and animal genomes have been sequenced, most centromere sequences have not been completed or analyzed in depth. To shed light on the unique organization, variability, and evolution of centromeric DNA, detailed analysis of a 1.97-Mb sequence that includes centromere 8 (CEN8) of japonica rice was undertaken. Thirty-three long-terminal repeat (LTR) retrotransposon families (including 11 previously unknown) were identified in the CEN8 region, totaling 245 elements and fragments that account for 67% of the region. The ratio of solo LTRs to intact elements in the CEN8 region is approximately 0.9:1, compared with approximately 2.2:1 in noncentromeric regions of rice. However, the ratio of solo LTRs to intact elements in the core of the CEN8 region ( approximately 2.5:1) is higher than in any other region investigated in rice, suggesting a hotspot for unequal recombination. Comparison of the CEN8 region of japonica and its orthologous segments from indica rice indicated that approximately 15% of the intact retrotransposons and solo LTRs were inserted into CEN8 after the divergence of japonica and indica from a common ancestor, compared with approximately 50% for previously studied euchromatic regions. Frequent DNA rearrangements were observed in the CEN8 region, including a 212-kb subregion that was found to be composed of three rearranged tandem repeats. Phylogenetic analysis also revealed recent segmental duplication and extensive rearrangement and reshuffling of the CentO satellite repeats. PMID- 16381820 TI - The up-regulation of ferritin expression using a small-molecule ligand to the native mRNA. AB - The binding of small molecules to distinctive three-dimensional structures in mRNA provides a new dimension in RNA control, previously limited to the targeting of secondary structures with antisense and RNA interference; such targeting can modulate mRNA function and rates of protein biosynthesis. Small molecules that selectively bind the iron-responsive element (IRE), a specific three-dimensional structure in the noncoding region of the ferritin mRNA model that is recognized by the iron-regulatory protein repressor, were identified by using chemical footprinting. The assay used involved an oxoruthenium(IV) complex that oxidizes guanine bases in RNA sequences. Small molecules that blocked oxidation of guanines in the internal loop region were expected to selectively increase the rate of ferritin synthesis, because the internal loop region of the ferritin IRE is distinctive from those of other IREs. The natural product yohimbine was found (based on gel mobility shifts) to block cleavage of the internal loop RNA site by >50% and seemed to inhibit protein binding. In the presence of yohimbine, the rate of biosynthesis of ferritin in a cell-free expression system (rabbit reticulocyte lysate) increased by 40%. Assignment of the IRE-yohimbine interaction as the origin of this effect was supported by a similar increase in synthesis of luciferase protein in a chimera of the IRE and luciferase gene. The identification of a small, drug-like molecule that recognizes a naturally occurring three-dimensional mRNA structure and regulates protein biosynthesis rates raises the possibility that small molecules can regulate protein biosynthesis by selectively binding to mRNA. PMID- 16381821 TI - Gene expression in human thyrocytes and autonomous adenomas reveals suppression of negative feedbacks in tumorigenesis. AB - The cAMP signaling pathway regulates growth of many cell types, including somatotrophs, thyrocytes, melanocytes, ovarian follicular granulosa cells, adrenocortical cells, and keratinocytes. Mutations of partners from the cAMP signaling cascade are involved in tumor formation. Thyroid-stimulating hormone (TSH) receptor and Gsalpha activating mutations have been detected in thyroid autonomous adenomas, Gsalpha mutations in growth hormone-secreting pituitary adenomas, and PKAR1A mutations in Carney complex, a multiple neoplasia syndrome. To gain more insight into the role of cAMP signaling in tumor formation, human primary cultures of thyrocytes were treated for different times (1.5, 3, 16, 24, and 48 h) with TSH to characterize modulations in gene expression using cDNA microarrays. This kinetic study showed a clear difference in expression, early (1.5 and 3 h) and late (16-48 h) after the onset of TSH stimulation. This result suggests a progressive sequential process leading to a change of cell program. The gene expression profile of the long-term stimulated cultures resembled the autonomous adenomas, but not papillary carcinomas. The molecular phenotype of the adenomas thus confirms the role of long-term stimulation of the TSH-cAMP cascade in the pathology. TSH induced a striking up-regulation of different negative feedback modulators of the cAMP cascade, presumably insuring the one-shot effect of the stimulus. Some were down- or nonregulated in adenomas, suggesting a loss of negative feedback control in the tumors. These results suggest that in tumorigenesis, activation of proliferation pathways may be complemented by suppression of multiple corresponding negative feedbacks, i.e., specific tumor suppressors. PMID- 16381823 TI - EMBL Nucleotide Sequence Database: developments in 2005. AB - The EMBL Nucleotide Sequence Database (www.ebi.ac.uk/embl) at the EMBL European Bioinformatics Institute, UK, offers a comprehensive set of publicly available nucleotide sequence and annotation, freely accessible to all. Maintained in collaboration with partners DDBJ and GenBank, coverage includes whole genome sequencing project data, directly submitted sequence, sequence recorded in support of patent applications and much more. The database continues to offer submission tools, data retrieval facilities and user support. In 2005, the volume of data offered has continued to grow exponentially. In addition to the newly presented data, the database encompasses a range of new data types generated by novel technologies, offers enhanced presentation and searchability of the data and has greater integration with other data resources offered at the EBI and elsewhere. In stride with these developing data types, the database has continued to develop submission and retrieval tools to maximise the information content of submitted data and to offer the simplest possible submission routes for data producers. New developments, the submission process, data retrieval and access to support are presented in this paper, along with links to sources of further information. PMID- 16381824 TI - TiProD: the Tissue-specific Promoter Database. AB - TiProD is a database of human promoter sequences for which some functional features are known. It allows a user to query individual promoters and the expression pattern they mediate, gene expression signatures of individual tissues, and to retrieve sets of promoters according to their tissue-specific activity or according to individual Gene Ontology terms the corresponding genes are assigned to. We have defined a measure for tissue-specificity that allows the user to discriminate between ubiquitously and specifically expressed genes. The database is accessible at http://tiprod.cbi.pku.edu.cn:8080/index.html. PMID- 16381825 TI - TRANSFAC and its module TRANSCompel: transcriptional gene regulation in eukaryotes. AB - The TRANSFAC database on transcription factors, their binding sites, nucleotide distribution matrices and regulated genes as well as the complementing database TRANSCompel on composite elements have been further enhanced on various levels. A new web interface with different search options and integrated versions of Match and Patch provides increased functionality for TRANSFAC. The list of databases which are linked to the common GENE table of TRANSFAC and TRANSCompel has been extended by: Ensembl, UniGene, EntrezGene, HumanPSD and TRANSPRO. Standard gene names from HGNC, MGI and RGD, are included for human, mouse and rat genes, respectively. With the help of InterProScan, Pfam, SMART and PROSITE domains are assigned automatically to the protein sequences of the transcription factors. TRANSCompel contains now, in addition to the COMPEL table, a separate table for detailed information on the experimental EVIDENCE on which the composite elements are based. Finally, for TRANSFAC, in respect of data growth, in particular the gain of Drosophila transcription factor binding sites (by courtesy of the Drosophila DNase I footprint database) and of Arabidopsis factors (by courtesy of DATF, Database of Arabidopsis Transcription Factors) has to be stressed. The here described public releases, TRANSFAC 7.0 and TRANSCompel 7.0, are accessible under http://www.gene-regulation.com/pub/databases.html. PMID- 16381826 TI - ARED 3.0: the large and diverse AU-rich transcriptome. AB - A comprehensive search that utilized a large set of mRNA data from human genome databases and additionally, expressed sequence tag (EST) database characterized this latest update of AU-rich elements (AREs) containing mRNA database (ARED). A large number of ARE-mRNA, as much as 4000, were recovered and include many of ARE alternative forms. This number represents as much as 5-8% of the human genes depending on the entire number of genes. The new ARED does not contain only larger and diverse number of ARE-mRNAs but additional functionality and enhanced search capabilities are given in the database website http://rc.kfshrc.edu.sa/ared/. These include class and cluster of AREs, source mRNAs, EST evidence, buildup information, retrieval of lists of genes, and integration with current and new NCBI data, such as Entrez ID and Unigene. Gene Ontology analysis shows there are significant differences in functional diversity of ARED when compared with the overall genome. Many of ARE-genes mediate regulatory processes, reactions to outside stimuli, RNA metabolism, and developmental processes particularly those of early and transient responses. The wide interest in mRNA turnover and importance of AREs in health and disease signify the compilation of ARE-genes. PMID- 16381827 TI - Argonaute--a database for gene regulation by mammalian microRNAs. AB - MicroRNAs (miRNAs) constitute a recently discovered class of small non-coding RNAs that regulate expression of target genes either by decreasing the stability of the target mRNA or by translational inhibition. They are involved in diverse processes, including cellular differentiation, proliferation and apoptosis. Recent evidence also suggests their importance for cancerogenesis. By far the most important model systems in cancer research are mammalian organisms. Thus, we decided to compile comprehensive information on mammalian miRNAs, their origin and regulated target genes in an exhaustive, curated database called Argonaute (http://www.ma.uni-heidelberg.de/apps/zmf/argonaute/interface). Argonaute collects latest information from both literature and other databases. In contrast to current databases on miRNAs like miRBase::Sequences, NONCODE or RNAdb, Argonaute hosts additional information on the origin of an miRNA, i.e. in which host gene it is encoded, its expression in different tissues and its known or proposed function, its potential target genes including Gene Ontology annotation, as well as miRNA families and proteins known to be involved in miRNA processing. Additionally, target genes are linked to an information retrieval system that provides comprehensive information from sequence databases and a simultaneous search of MEDLINE with all synonyms of a given gene. The web interface allows the user to get information for a single or multiple miRNAs, either selected or uploaded through a text file. Argonaute currently has information on 839 miRNAs from human, mouse and rat. PMID- 16381828 TI - GRSDB: a database of quadruplex forming G-rich sequences in alternatively processed mammalian pre-mRNA sequences. AB - Guanine-rich nucleic acids are known to form highly stable G-quadruplex structures, also known as G-quartets. Recently, there has been a tremendous amount of interest in studying G-quadruplexes owing to the realization of their biological importance. G-rich sequences (GRSs) capable of forming G-quadruplexes are found in the vicinity of polyadenylation regions and are involved in regulating 3' end processing of mammalian pre-mRNAs. G-rich motifs are also known to play an important role in alternative, tissue-specific splicing by interacting with hnRNP H protein subfamily. Whether quadruplex structure directly plays a role in regulating RNA processing events requires further investigation. To date there has not been a comprehensive effort to study G-quadruplexes near RNA processing sites. We have applied a computational approach to map putative Quadruplex forming GRSs within the transcribed regions of a large number of alternatively processed human and mouse gene sequences that were obtained as fully annotated entries from GenBank and RefSeq. We have used the computed data to build the GRSDB database that provides a unique avenue for studying G quadruplexes in the context of RNA processing sites. GRSDB website offers visual comparison of G-quadruplex distribution patterns among all the alternative RNA products of a gene with the help of dynamic graphics. At present, GRSDB contains data from 1310 human and mouse genes, of which 1188 are alternatively processed. It has a total of 379,223 predicted G-quadruplexes, of which 54,252 are near RNA processing sites. GRSDB is a good resource for researchers interested in investigating the functional relevance of G-quadruplexes, especially in the context of alternative RNA processing. It can be accessed at http://bioinformatics.ramapo.edu/grsdb/. PMID- 16381829 TI - IRESite: the database of experimentally verified IRES structures (www.iresite.org). AB - IRESite is an exhaustive, manually annotated non-redundant relational database focused on the IRES elements (Internal Ribosome Entry Site) and containing information not available in the primary public databases. IRES elements were originally found in eukaryotic viruses hijacking initiation of translation of their host. Later on, they were also discovered in 5'-untranslated regions of some eukaryotic mRNA molecules. Currently, IRESite presents up to 92 biologically relevant aspects of every experiment, e.g. the nature of an IRES element, its functionality/defectivity, origin, size, sequence, structure, its relative position with respect to surrounding protein coding regions, positive/negative controls used in the experiment, the reporter genes used to monitor IRES activity, the measured reporter protein yields/activities, and references to original publications as well as cross-references to other databases, and also comments from submitters and our curators. Furthermore, the site presents the known similarities to rRNA sequences as well as RNA-protein interactions. Special care is given to the annotation of promoter-like regions. The annotated data in IRESite are bound to mostly complete, full-length mRNA, and whenever possible, accompanied by original plasmid vector sequences. New data can be submitted through the publicly available web-based interface at http://www.iresite.org and are curated by a team of lab-experienced biologists. PMID- 16381830 TI - MeRNA: a database of metal ion binding sites in RNA structures. AB - Metal ions are essential for the folding of RNA into stable tertiary structures and for the catalytic activity of some RNA enzymes. To aid in the study of the roles of metal ions in RNA structural biology, we have created MeRNA (Metals in RNA), a comprehensive compilation of all metal binding sites identified in RNA 3D structures available from the PDB and Nucleic Acid Database. Currently, our database contains information relating to binding of 9764 metal ions corresponding to 23 distinct elements, in 256 RNA structures. The metal ion locations were confirmed and ligands characterized using original literature references. MeRNA includes eight manually identified metal-ion binding motifs, which are described in the literature. MeRNA is searchable by PDB identifier, metal ion, method of structure determination, resolution and R-values for X-ray structure and distance from metal to any RNA atom or to water. New structures with their respective binding motifs will be added to the database as they become available. The MeRNA database will further our understanding of the roles of metal ions in RNA folding and catalysis and have applications in structural and functional analysis, RNA design and engineering. The MeRNA database is accessible at http://merna.lbl.gov. PMID- 16381831 TI - miRNAMap: genomic maps of microRNA genes and their target genes in mammalian genomes. AB - Recent work has demonstrated that microRNAs (miRNAs) are involved in critical biological processes by suppressing the translation of coding genes. This work develops an integrated database, miRNAMap, to store the known miRNA genes, the putative miRNA genes, the known miRNA targets and the putative miRNA targets. The known miRNA genes in four mammalian genomes such as human, mouse, rat and dog are obtained from miRBase, and experimentally validated miRNA targets are identified in a survey of the literature. Putative miRNA precursors were identified by RNAz, which is a non-coding RNA prediction tool based on comparative sequence analysis. The mature miRNA of the putative miRNA genes is accurately determined using a machine learning approach, mmiRNA. Then, miRanda was applied to predict the miRNA targets within the conserved regions in 3'-UTR of the genes in the four mammalian genomes. The miRNAMap also provides the expression profiles of the known miRNAs, cross-species comparisons, gene annotations and cross-links to other biological databases. Both textual and graphical web interface are provided to facilitate the retrieval of data from the miRNAMap. The database is freely available at http://mirnamap.mbc.nctu.edu.tw/. PMID- 16381832 TI - miRBase: microRNA sequences, targets and gene nomenclature. AB - The miRBase database aims to provide integrated interfaces to comprehensive microRNA sequence data, annotation and predicted gene targets. miRBase takes over functionality from the microRNA Registry and fulfils three main roles: the miRBase Registry acts as an independent arbiter of microRNA gene nomenclature, assigning names prior to publication of novel miRNA sequences. miRBase Sequences is the primary online repository for miRNA sequence data and annotation. miRBase Targets is a comprehensive new database of predicted miRNA target genes. miRBase is available at http://microrna.sanger.ac.uk/. PMID- 16381833 TI - MODOMICS: a database of RNA modification pathways. AB - MODOMICS is the first comprehensive database resource for systems biology of RNA modification. It integrates information about the chemical structure of modified nucleosides, their localization in RNA sequences, pathways of their biosynthesis and enzymes that carry out the respective reactions. MODOMICS also provides literature information, and links to other databases, including the available protein sequence and structure data. The current list of modifications and pathways is comprehensive, while the dataset of enzymes is limited to Escherichia coli and Saccharomyces cerevisiae and sequence alignments are presented only for tRNAs from these organisms. RNAs and enzymes from other organisms will be included in the near future. MODOMICS can be queried by the type of nucleoside (e.g. A, G, C, U, I, m1A, nm5s2U, etc.), type of RNA, position of a particular nucleoside, type of reaction (e.g. methylation, thiolation, deamination, etc.) and name or sequence of an enzyme of interest. Options for data presentation include graphs of pathways involving the query nucleoside, multiple sequence alignments of RNA sequences and tabular forms with enzyme and literature data. The contents of MODOMICS can be accessed through the World Wide Web at http://genesilico.pl/modomics/. PMID- 16381834 TI - NPInter: the noncoding RNAs and protein related biomacromolecules interaction database. AB - The noncoding RNAs and protein related biomacromolecules interaction database (NPInter; http://bioinfo.ibp.ac.cn/NPInter or http://www.bioinfo.org.cn/NPInter) is a database that documents experimentally determined functional interactions between noncoding RNAs (ncRNAs) and protein related biomacromolecules (PRMs) (proteins, mRNAs or genomic DNAs). NPInter intends to provide the scientific community with a comprehensive and integrated tool for efficient browsing and extraction of information on interactions between ncRNAs and PRMs. Beyond cataloguing details of these interactions, the NPInter will be useful for understanding ncRNA function, as it adds a very important functional element, ncRNAs, to the biomolecule interaction network and sets up a bridge between the coding and the noncoding kingdoms. PMID- 16381835 TI - RNAi Codex: a portal/database for short-hairpin RNA (shRNA) gene-silencing constructs. AB - Use of RNA interference (RNAi) in forward genetic screens is proliferating. Currently, short-interfering RNAs (siRNAs) and short-hairpin RNAs (shRNAs) are being used to silence genes to tease out functional information. It is becoming easier to harness RNAi to silence specific genes, owing to the development of libraries of readymade shRNA and siRNA gene-silencing constructs by using a variety of sources. RNAi Codex, which consists of a database of shRNA related information and an associated website, has been developed as a portal for publicly available shRNA resources and is accessible at http://codex.cshl.org. RNAi Codex currently holds data from the Hannon-Elledge shRNA library and allows the use of biologist-friendly gene names to access information on shRNA constructs that can silence the gene of interest. It is designed to hold user contributed annotations and publications for each construct, as and when such data become available. We will describe features of RNAi Codex and explain the use of the tool. PMID- 16381836 TI - snoRNA-LBME-db, a comprehensive database of human H/ACA and C/D box snoRNAs. AB - The snoRNA-LBME-db is a dedicated database containing human C/D box and H/ACA box small nucleolar RNAs (snoRNAs), and small Cajal body-specific RNAs (scaRNAs). C/D box and H/ACA box snoRNAs are part of ribonucleoparticles that guide 2'-O-ribose methylation and pseudouridilation, respectively, of selected residues of 28S, 18S or 5.8S rRNAs or of the spliceosomal U6 RNA. Similarly, scaRNAs guide modifications of the spliceosomal RNAs transcribed by RNA polymerase II (U1, U2, U4, U5 and U12) and are often composed of both C/D box and H/ACA box domains. However, some snoRNAs do not function as modification guide RNAs, but rather as RNA chaperones during the maturation of pre-rRNA. The database was built by a compilation of the literature, and comprises human sno/scaRNAs that were experimentally verified, as well as the human orthologs of snoRNAs that were cloned in other vertebrate species, and some snoRNAs that are predicted by bioinformatics search in loci submitted to genomic imprinting, but have not all been experimentally verified. For each entry, the database identifies the modified nucleotide(s) in the target RNA(s), indicates the corresponding predicted base pairing, gives a few pertinent references and provides a link to the position of the sno/scaRNA on the UCSC Genome Browser. The 'Find guide RNA' function allows one to find the sno/scaRNAs predicted to guide the modification of a particular nucleotide in the rRNA and spliceosomal RNA sequences. The 'Browse' function allows one to download the sequences of selected sno/scaRNAs in the FASTA format. The database is available online at http://www snorna.biotoul.fr/. It can also be accessed from the human UCSC Genome Browser via the sno/miRNA track. PMID- 16381837 TI - GenBank. AB - GenBank (R) is a comprehensive database that contains publicly available DNA sequences for more than 205 000 named organisms, obtained primarily through submissions from individual laboratories and batch submissions from large-scale sequencing projects. Most submissions are made using the Web-based BankIt or standalone Sequin programs and accession numbers are assigned by GenBank staff upon receipt. Daily data exchange with the EMBL Data Library in Europe and the DNA Data Bank of Japan ensures worldwide coverage. GenBank is accessible through NCBI's retrieval system, Entrez, which integrates data from the major DNA and protein sequence databases along with taxonomy, genome, mapping, protein structure and domain information, and the biomedical journal literature via PubMed. BLAST provides sequence similarity searches of GenBank and other sequence databases. Complete bimonthly releases and daily updates of the GenBank database are available by FTP. To access GenBank and its related retrieval and analysis services, go to the NCBI Homepage at www.ncbi.nlm.nih.gov. PMID- 16381838 TI - The tmRDB and SRPDB resources. AB - Maintained at the University of Texas Health Science Center at Tyler, Texas, the tmRNA database (tmRDB) is accessible at the URL http://psyche.uthct.edu/dbs/tmRDB/tmRDB.html with mirror sites located at Auburn University, Auburn, Alabama (http://www.ag.auburn.edu/mirror/tmRDB/) and the Royal Veterinary and Agricultural University, Denmark (http://tmrdb.kvl.dk/). The signal recognition particle database (SRPDB) at http://psyche.uthct.edu/dbs/SRPDB/SRPDB.html is mirrored at http://srpdb.kvl.dk/ and the University of Goteborg (http://bio.lundberg.gu.se/dbs/SRPDB/SRPDB.html). The databases assist in investigations of the tmRNP (a ribonucleoprotein complex which liberates stalled bacterial ribosomes) and the SRP (a particle which recognizes signal sequences and directs secretory proteins to cell membranes). The curated tmRNA and SRP RNA alignments consider base pairs supported by comparative sequence analysis. Also shown are alignments of the tmRNA-associated proteins SmpB, ribosomal protein S1, alanyl-tRNA synthetase and Elongation Factor Tu, as well as the SRP proteins SRP9, SRP14, SRP19, SRP21, SRP54 (Ffh), SRP68, SRP72, cpSRP43, Flhf, SRP receptor (alpha) and SRP receptor (beta). All alignments can be easily examined using a new exploratory browser. The databases provide links to high-resolution structures and serve as depositories for structures obtained by molecular modeling. PMID- 16381839 TI - MIPS: analysis and annotation of proteins from whole genomes in 2005. AB - The Munich Information Center for Protein Sequences (MIPS at the GSF), Neuherberg, Germany, provides resources related to genome information. Manually curated databases for several reference organisms are maintained. Several of these databases are described elsewhere in this and other recent NAR database issues. In a complementary effort, a comprehensive set of >400 genomes automatically annotated with the PEDANT system are maintained. The main goal of our current work on creating and maintaining genome databases is to extend gene centered information to information on interactions within a generic comprehensive framework. We have concentrated our efforts along three lines (i) the development of suitable comprehensive data structures and database technology, communication and query tools to include a wide range of different types of information enabling the representation of complex information such as functional modules or networks Genome Research Environment System, (ii) the development of databases covering computable information such as the basic evolutionary relations among all genes, namely SIMAP, the sequence similarity matrix and the CABiNet network analysis framework and (iii) the compilation and manual annotation of information related to interactions such as protein-protein interactions or other types of relations (e.g. MPCDB, MPPI, CYGD). All databases described and the detailed descriptions of our projects can be accessed through the MIPS WWW server (http://mips.gsf.de). PMID- 16381840 TI - Database resources of the National Center for Biotechnology Information. AB - In addition to maintaining the GenBank nucleic acid sequence database, the National Center for Biotechnology Information (NCBI) provides analysis and retrieval resources for the data in GenBank and other biological data made available through NCBI's Web site. NCBI resources include Entrez, the Entrez Programming Utilities, MyNCBI, PubMed, PubMed Central, Entrez Gene, the NCBI Taxonomy Browser, BLAST, BLAST Link (BLink), Electronic PCR, OrfFinder, Spidey, Splign, RefSeq, UniGene, HomoloGene, ProtEST, dbMHC, dbSNP, Cancer Chromosomes, Entrez Genomes and related tools, the Map Viewer, Model Maker, Evidence Viewer, Clusters of Orthologous Groups, Retroviral Genotyping Tools, HIV-1, Human Protein Interaction Database, SAGEmap, Gene Expression Omnibus, Entrez Probe, GENSAT, Online Mendelian Inheritance in Man, Online Mendelian Inheritance in Animals, the Molecular Modeling Database, the Conserved Domain Database, the Conserved Domain Architecture Retrieval Tool and the PubChem suite of small molecule databases. Augmenting many of the Web applications are custom implementations of the BLAST program optimized to search specialized datasets. All of the resources can be accessed through the NCBI home page at: http://www.ncbi.nlm.nih.gov. PMID- 16381841 TI - TCDB: the Transporter Classification Database for membrane transport protein analyses and information. AB - The Transporter Classification Database (TCDB) is a web accessible, curated, relational database containing sequence, classification, structural, functional and evolutionary information about transport systems from a variety of living organisms. TCDB is a curated repository for factual information compiled from >10,000 references, encompassing approximately 3000 representative transporters and putative transporters, classified into >400 families. The transporter classification (TC) system is an International Union of Biochemistry and Molecular Biology approved system of nomenclature for transport protein classification. TCDB is freely accessible at http://www.tcdb.org. The web interface provides several different methods for accessing the data, including step-by-step access to hierarchical classification, direct search by sequence or TC number and full-text searching. The functional ontology that underlies the database structure facilitates powerful query searches that yield valuable data in a quick and easy way. The TCDB website also offers several tools specifically designed for analyzing the unique characteristics of transport proteins. TCDB not only provides curated information and a tool for classifying newly identified membrane proteins, but also serves as a genome transporter-annotation tool. PMID- 16381842 TI - The Universal Protein Resource (UniProt): an expanding universe of protein information. AB - The Universal Protein Resource (UniProt) provides a central resource on protein sequences and functional annotation with three database components, each addressing a key need in protein bioinformatics. The UniProt Knowledgebase (UniProtKB), comprising the manually annotated UniProtKB/Swiss-Prot section and the automatically annotated UniProtKB/TrEMBL section, is the preeminent storehouse of protein annotation. The extensive cross-references, functional and feature annotations and literature-based evidence attribution enable scientists to analyse proteins and query across databases. The UniProt Reference Clusters (UniRef) speed similarity searches via sequence space compression by merging sequences that are 100% (UniRef100), 90% (UniRef90) or 50% (UniRef50) identical. Finally, the UniProt Archive (UniParc) stores all publicly available protein sequences, containing the history of sequence data with links to the source databases. UniProt databases continue to grow in size and in availability of information. Recent and upcoming changes to database contents, formats, controlled vocabularies and services are described. New download availability includes all major releases of UniProtKB, sequence collections by taxonomic division and complete proteomes. A bibliography mapping service has been added, and an ID mapping service will be available soon. UniProt databases can be accessed online at http://www.uniprot.org or downloaded at ftp://ftp.uniprot.org/pub/databases/. PMID- 16381843 TI - CyBase: a database of cyclic protein sequence and structure. AB - CyBase is a curated database and information source for backbone-cyclized proteins. The database incorporates naturally occurring cyclic proteins as well as synthetic derivatives, grafted analogues and acyclic permutants. The database provides a centralized repository of information on all aspects of cyclic protein biology and addresses issues pertaining to the management and searching of topologically circular sequences. The database is freely available at http://research.imb.uq.edu.au/cybase. PMID- 16381844 TI - PINT: Protein-protein Interactions Thermodynamic Database. AB - The first release of Protein-protein Interactions Thermodynamic Database (PINT) contains >1500 data of several thermodynamic parameters along with sequence and structural information, experimental conditions and literature information. Each entry contains numerical data for the free energy change, dissociation constant, association constant, enthalpy change, heat capacity change and so on of the interacting proteins upon binding, which are important for understanding the mechanism of protein-protein interactions. PINT also includes the name and source of the proteins involved in binding, their Protein Information Resource, SWISS PROT and Protein Data Bank (PDB) codes, secondary structure and solvent accessibility of residues at mutant positions, measuring methods, experimental conditions, such as buffers, ions and additives, and literature information. A WWW interface facilitates users to search data based on various conditions, feasibility to select the terms for output and different sorting options. Further, PINT is cross-linked with other related databases, PIR, SWISS-PROT, PDB and NCBI PUBMED literature database. The database is freely available at http://www.bioinfodatabase.com/pint/index.html. PMID- 16381845 TI - PPD v1.0--an integrated, web-accessible database of experimentally determined protein pKa values. AB - The Protein pK(a) Database (PPD) v1.0 provides a compendium of protein residue specific ionization equilibria (pK(a) values), as collated from the primary literature, in the form of a web-accessible postgreSQL relational database. Ionizable residues play key roles in the molecular mechanisms that underlie many biological phenomena, including protein folding and enzyme catalysis. The PPD serves as a general protein pK(a) archive and as a source of data that allows for the development and improvement of pK(a) prediction systems. The database is accessed through an HTML interface, which offers two fast, efficient search methods: an amino acid-based query and a Basic Local Alignment Search Tool search. Entries also give details of experimental techniques and links to other key databases, such as National Center for Biotechnology Information and the Protein Data Bank, providing the user with considerable background information. The database can be found at the following URL: http://www.jenner.ac.uk/PPD. PMID- 16381846 TI - ProTherm and ProNIT: thermodynamic databases for proteins and protein-nucleic acid interactions. AB - ProTherm and ProNIT are two thermodynamic databases that contain experimentally determined thermodynamic parameters of protein stability and protein-nucleic acid interactions, respectively. The current versions of both the databases have considerably increased the total number of entries and enhanced search interface with added new fields, improved search, display and sorting options. As on September 2005, ProTherm release 5.0 contains 17,113 entries from 771 proteins, retrieved from 1497 scientific articles (approximately 20% increase in data from the previous version). ProNIT release 2.0 contains 4900 entries from 273 research articles, representing 158 proteins. Both databases can be queried using WWW interfaces. Both quick search and advanced search are provided on this web page to facilitate easy retrieval and display of the data from these databases. ProTherm is freely available online at http://gibk26.bse.kyutech.ac.jp/jouhou/Protherm/protherm.html and ProNIT at http://gibk26.bse.kyutech.ac.jp/jouhou/pronit/pronit.html. PMID- 16381847 TI - The REFOLD database: a tool for the optimization of protein expression and refolding. AB - A large proportion of proteins expressed in Escherichia coli form inclusion bodies and thus require renaturation to attain a functional conformation for analysis. In this process, identifying and optimizing the refolding conditions and methodology is often rate limiting. In order to address this problem, we have developed REFOLD, a web-accessible relational database containing the published methods employed in the refolding of recombinant proteins. Currently, REFOLD contains >300 entries, which are heavily annotated such that the database can be searched via multiple parameters. We anticipate that REFOLD will continue to grow and eventually become a powerful tool for the optimization of protein renaturation. REFOLD is freely available at http://refold.med.monash.edu.au. PMID- 16381848 TI - ChimerDB--a knowledgebase for fusion sequences. AB - Chromosome translocation and gene fusion are frequent events in the human genome and are often the cause of many types of tumor. ChimerDB is the database of fusion sequences encompassing bioinformatics analysis of mRNA and expressed sequence tag (EST) sequences in the GenBank, manual collection of literature data and integration with other known database such as OMIM. Our bioinformatics analysis identifies the fusion transcripts that have non-overlapping alignments at multiple genomic loci. Fusion events at exon-exon borders are selected to filter out the cloning artifacts in cDNA library preparation. The result is classified into two groups--genuine chromosome translocation and fusion between neighboring genes owing to intergenic splicing. We also integrated manually collected literature and OMIM data for chromosome translocation as an aid to assess the validity of each fusion event. The database is available at http://genome.ewha.ac.kr/ChimerDB/ for human, mouse and rat genomes. PMID- 16381849 TI - LOCATE: a mouse protein subcellular localization database. AB - We present here LOCATE, a curated, web-accessible database that houses data describing the membrane organization and subcellular localization of proteins from the FANTOM3 Isoform Protein Sequence set. Membrane organization is predicted by the high-throughput, computational pipeline MemO. The subcellular locations of selected proteins from this set were determined by a high-throughput, immunofluorescence-based assay and by manually reviewing >1700 peer-reviewed publications. LOCATE represents the first effort to catalogue the experimentally verified subcellular location and membrane organization of mammalian proteins using a high-throughput approach and provides localization data for approximately 40% of the mouse proteome. It is available at http://locate.imb.uq.edu.au. PMID- 16381850 TI - NOPdb: Nucleolar Proteome Database. AB - The Nucleolar Proteome Database (NOPdb) archives data on >700 proteins that were identified by multiple mass spectrometry (MS) analyses from highly purified preparations of human nucleoli, the most prominent nuclear organelle. Each protein entry is annotated with information about its corresponding gene, its domain structures and relevant protein homologues across species, as well as documenting its MS identification history including all the peptides sequenced by tandem MS/MS. Moreover, data showing the quantitative changes in the relative levels of approximately 500 nucleolar proteins are compared at different timepoints upon transcriptional inhibition. Correlating changes in protein abundance at multiple timepoints, highlighted by visualization means in the NOPdb, provides clues regarding the potential interactions and relationships between nucleolar proteins and thereby suggests putative functions for factors within the 30% of the proteome which comprises novel/uncharacterized proteins. The NOPdb (http://www.lamondlab.com/NOPdb) is searchable by either gene names, nucleotide or protein sequences, Gene Ontology terms or motifs, or by limiting the range for isoelectric points and/or molecular weights and links to other databases (e.g. LocusLink, OMIM and PubMed). PMID- 16381851 TI - Nuclear Receptor Signaling Atlas (www.nursa.org): hyperlinking the nuclear receptor signaling community. AB - The nuclear receptor signaling (NRS) field has generated a substantial body of information on nuclear receptors, their ligands and coregulators, with the ultimate goal of constructing coherent models of the biological and clinical significance of these molecules. As a component of the Nuclear Receptor Signaling Atlas (NURSA)--the development of a functional atlas of nuclear receptor biology- the NURSA Bioinformatics Resource is developing a strategy to organize and integrate legacy and future information on these molecules in a single web-based resource (www.nursa.org). This entails parallel efforts of (i) developing an appropriate software framework for handling datasets from NURSA laboratories and (ii) designing strategies for the curation and presentation of public data relevant to NRS. To illustrate our approach, we have described here in detail the development of a web-based interface for the NURSA quantitative PCR nuclear receptor expression dataset, incorporating bioinformatics analysis which provides novel perspectives on functional relationships between these molecules. We anticipate that the free and open access of the community to a platform for data mining and hypothesis generation strategies will be a significant contribution to the progress of research in this field. PMID- 16381852 TI - The PROSITE database. AB - The PROSITE database consists of a large collection of biologically meaningful signatures that are described as patterns or profiles. Each signature is linked to a documentation that provides useful biological information on the protein family, domain or functional site identified by the signature. The PROSITE database is now complemented by a series of rules that can give more precise information about specific residues. During the last 2 years, the documentation and the ScanProsite web pages were redesigned to add more functionalities. The latest version of PROSITE (release 19.11 of September 27, 2005) contains 1329 patterns and 552 profile entries. Over the past 2 years more than 200 domains have been added, and now 52% of UniProtKB/Swiss-Prot entries (release 48.1 of September 27, 2005) have a cross-reference to a PROSITE entry. The database is accessible at http://www.expasy.org/prosite/. PMID- 16381853 TI - SitesBase: a database for structure-based protein-ligand binding site comparisons. AB - There are many components which govern the function of a protein within a cell. Here, we focus on the molecular recognition of small molecules and the prediction of common recognition by similarity between protein-ligand binding sites. SitesBase is an easily accessible database which is simple to use and holds information about structural similarities between known ligand binding sites found in the Protein Data Bank. These similarities are presented to the wider community enabling full analysis of molecular recognition and potentially protein structure-function relationships. SitesBase is accessible at http://www.bioinformatics.leeds.ac.uk/sb. PMID- 16381854 TI - BIOZON: a hub of heterogeneous biological data. AB - Biological entities are strongly related and mutually dependent on each other. Therefore, there is a growing need to corroborate and integrate data from different resources and aspects of biological systems in order to analyze them effectively. Biozon is a unified biological database that integrates heterogeneous data types such as proteins, structures, domain families, protein protein interactions and cellular pathways, and establishes the relationships between them. All data are integrated on to a single graph schema centered around the non-redundant set of biological objects that are shared by each source. This integration results in a highly connected graph structure that provides a more complete picture of the known context of a given object that cannot be determined from any one source. Currently, Biozon integrates roughly 2 million protein sequences, 42 million DNA or RNA sequences, 32,000 protein structures, 150,000 interactions and more from sources such as GenBank, UniProt, Protein Data Bank (PDB) and BIND. Biozon augments source data with locally derived data such as 5 billion pairwise protein alignments and 8 million structural alignments. The user may form complex cross-type queries on the graph structure, add similarity relations to form fuzzy queries and rank the results based on analysis of the edge structure similar to Google PageRank, online at Biozon.org. PMID- 16381855 TI - MulPSSM: a database of multiple position-specific scoring matrices of protein domain families. AB - Representation of multiple sequence alignments of protein families in terms of position-specific scoring matrices (PSSMs) is commonly used in the detection of remote homologues. A PSSM is generated with respect to one of the sequences involved in the multiple sequence alignment as a reference. We have shown recently that the use of multiple PSSMs corresponding to an alignment, with several sequences in the family used as reference, improves the sensitivity of the remote homology detection dramatically. MulPSSM contains PSSMs for a large number of sequence and structural families of protein domains with multiple PSSMs for every family. The approach involves use of a clustering algorithm to identify most distinct sequences corresponding to a family. With each one of the distinct sequences as reference, multiple PSSMs have been generated. The current release of MulPSSM contains approximately 33,000 and approximately 38,000 PSSMs corresponding to 7868 sequence and 2625 structural families. A RPS_BLAST interface allows sequence search against PSSMs of sequence or structural families or both. An analysis interface allows display and convenient navigation of alignments and domain hits. MulPSSM can be accessed at http://crick.mbu.iisc.ernet.in/~mulpssm. PMID- 16381856 TI - Pfam: clans, web tools and services. AB - Pfam is a database of protein families that currently contains 7973 entries (release 18.0). A recent development in Pfam has enabled the grouping of related families into clans. Pfam clans are described in detail, together with the new associated web pages. Improvements to the range of Pfam web tools and the first set of Pfam web services that allow programmatic access to the database and associated tools are also presented. Pfam is available on the web in the UK (http://www.sanger.ac.uk/Software/Pfam/), the USA (http://pfam.wustl.edu/), France (http://pfam.jouy.inra.fr/) and Sweden (http://pfam.cgb.ki.se/). PMID- 16381857 TI - HUMHOT: a database of human meiotic recombination hot spots. AB - Meiotic recombination occurs preferentially at certain regions in the genome referred to as hot spots. The number of hot spots known in humans has increased manifold in recent years. The identification of these hot spots in humans is of great interest to population and medical geneticists since they influence the structure of Linkage Disequilibrium and Haplotype blocks in human populations, whose patterns have applications in mapping disease genes. HUMHOT is a web-based database of Human Meiotic Recombination Hot Spots. The database comprises DNA sequences corresponding to the hot spot regions from the literature that have been mapped to a high resolution (<4 kb) in humans. It also provides flanking sequence information for the hot spot region along with references describing the hot spot. The database can be queried based on hot spot identity, chromosome position or by homology to user-defined sequences. It is also updated with new hot spot sequences as they are discovered and provides hyperlinks to commonly used tools for estimating recombination rates, performing genetic analysis and new advances in our understanding of meiotic hot spots. Public access to the HUMHOT database is available at http://www.jncasr.ac.in/humhot. PMID- 16381858 TI - SIMAP: the similarity matrix of proteins. AB - Similarity Matrix of Proteins (SIMAP) (http://mips.gsf.de/simap) provides a database based on a pre-computed similarity matrix covering the similarity space formed by >4 million amino acid sequences from public databases and completely sequenced genomes. The database is capable of handling very large datasets and is updated incrementally. For sequence similarity searches and pairwise alignments, we implemented a grid-enabled software system, which is based on FASTA heuristics and the Smith-Waterman algorithm. Our ProtInfo system allows querying by protein sequences covered by the SIMAP dataset as well as by fragments of these sequences, highly similar sequences and title words. Each sequence in the database is supplemented with pre-calculated features generated by detailed sequence analyses. By providing WWW interfaces as well as web-services, we offer the SIMAP resource as an efficient and comprehensive tool for sequence similarity searches. PMID- 16381859 TI - SMART 5: domains in the context of genomes and networks. AB - The Simple Modular Architecture Research Tool (SMART) is an online resource (http://smart.embl.de/) used for protein domain identification and the analysis of protein domain architectures. Many new features were implemented to make SMART more accessible to scientists from different fields. The new 'Genomic' mode in SMART makes it easy to analyze domain architectures in completely sequenced genomes. Domain annotation has been updated with a detailed taxonomic breakdown and a prediction of the catalytic activity for 50 SMART domains is now available, based on the presence of essential amino acids. Furthermore, intrinsically disordered protein regions can be identified and displayed. The network context is now displayed in the results page for more than 350 000 proteins, enabling easy analyses of domain interactions. PMID- 16381860 TI - The EROP-Moscow oligopeptide database. AB - Natural oligopeptides may regulate nearly all vital processes. To date, the chemical structures of nearly 6000 oligopeptides have been identified from >1000 organisms representing all the biological kingdoms. We have compiled the known physical, chemical and biological properties of these oligopeptides--whether synthesized on ribosomes or by non-ribosomal enzymes--and have constructed an internet-accessible database, EROP-Moscow (Endogenous Regulatory OligoPeptides), which resides at http://erop.inbi.ras.ru. This database enables users to perform rapid searches via many key features of the oligopeptides, and to carry out statistical analysis of all the available information. The database lists only those oligopeptides whose chemical structures have been completely determined (directly or by translation from nucleotide sequences). It provides extensive links with the Swiss-Prot-TrEMBL peptide-protein database, as well as with the PubMed biomedical bibliographic database. EROP-Moscow also contains data on many oligopeptides that are absent from other convenient databases, and is designed for extended use in classifying new natural oligopeptides and for production of novel peptide pharmaceuticals. PMID- 16381861 TI - LGICdb: a manually curated sequence database after the genomes. AB - Ligand-gated ion channels form transmembrane ionic pores controlled by the binding of chemicals. The LGICdb aims to be a non-redundant, manually curated resource offering access to the large number of subunits composing extracellularly activated ligand-gated ion channels, such as nicotinic, ATP, GABA and glutamate ionotropic receptors. Composed of more than 500 human curated entries, the XML native database has been relocated in 2004 to the EBI. Its facilities have been enhanced with a new search system, customized multiple sequence alignments and manipulation of protein structures (http://www.ebi.ac.uk/compneur-srv/LGICdb/). Despite the vast improvement of general sequence resources, the LGICdb still provide sequences unavailable elsewhere. PMID- 16381862 TI - MEROPS: the peptidase database. AB - Peptidases (proteolytic enzymes) and their natural, protein inhibitors are of great relevance to biology, medicine and biotechnology. The MEROPS database (http://merops.sanger.ac.uk) aims to fulfil the need for an integrated source of information about these proteins. The organizational principle of the database is a hierarchical classification in which homologous sets of proteins of interest are grouped into families and the homologous families are grouped in clans. The most important addition to the database has been newly written, concise text annotations for each peptidase family. Other forms of information recently added include highlighting of active site residues (or the replacements that render some homologues inactive) in the sequence displays and BlastP search results, dynamically generated alignments and trees at the peptidase or inhibitor level, and a curated list of human and mouse homologues that have been experimentally characterized as active. A new way to display information at taxonomic levels higher than species has been devised. In the Literature pages, references have been flagged to draw attention to particularly 'hot' topics. PMID- 16381863 TI - DMAPS: a database of multiple alignments for protein structures. AB - The database of multiple alignments for protein structures (DMAPS) provides instant access to pre-computed multiple structure alignments for all protein structure families in the Protein Data Bank (PDB). Protein structure families have been obtained from four distinct classification methods including SCOP, CATH, ENZYME and CE, and multiple structure alignments have been built for all families containing at least three members, using CE-MC software. Currently, multiple structure alignments are available for 3050 SCOP-, 3087 CATH-, 664 ENZYME- and 1707 CE-based families. A web-based query system has been developed to retrieve multiple alignments for these families using the PDB chain ID of any member of a family. Multiple alignments can be viewed or downloaded in six different formats, including JOY/html, TEXT, FASTA, PDB (superimposed coordinates), JOY/postscript and JOY/rtf. DMAPS is accessible online at http://bioinformatics.albany.edu/~dmaps. PMID- 16381864 TI - Flexible Structural Neighborhood--a database of protein structural similarities and alignments. AB - Protein structures are flexible, changing their shapes not only upon substrate binding, but also during evolution as a collective effect of mutations, deletions and insertions. A new generation of protein structure comparison algorithms allows for such flexibility; they go beyond identifying the largest common part between two proteins and find hinge regions and patterns of flexibility in protein families. Here we present a Flexible Structural Neighborhood (FSN), a database of structural neighbors of proteins deposited in PDB as seen by a flexible protein structure alignment program FATCAT, developed previously in our group. The database, searchable by a protein PDB code, provides lists of proteins with statistically significant structural similarity and on lower menu levels provides detailed alignments, interactive superposition of structures and positions of hinges that were identified in the comparison. While superficially similar to other structural protein alignment resources, FSN provides a unique resource to study not only protein structural similarity, but also how protein structures change. FSN is available from a server http://fatcat.burnham.org/fatcat/struct_neighbor and by direct links from the PDB database. PMID- 16381865 TI - Gene3D: modelling protein structure, function and evolution. AB - The Gene3D release 4 database and web portal (http://cathwww.biochem.ucl.ac.uk:8080/Gene3D) provide a combined structural, functional and evolutionary view of the protein world. It is focussed on providing structural annotation for protein sequences without structural representatives--including the complete proteome sets of over 240 different species. The protein sequences have also been clustered into whole-chain families so as to aid functional prediction. The structural annotation is generated using HMM models based on the CATH domain families; CATH is a repository for manually deduced protein domains. Amongst the changes from the last publication are: the addition of over 100 genomes and the UniProt sequence database, domain data from Pfam, metabolic pathway and functional data from COGs, KEGG and GO, and protein protein interaction data from MINT and BIND. The website has been rebuilt to allow more sophisticated querying and the data returned is presented in a clearer format with greater functionality. Furthermore, all data can be downloaded in a simple XML format, allowing users to carry out complex investigations at their own computers. PMID- 16381866 TI - iMOTdb--a comprehensive collection of spatially interacting motifs in proteins. AB - Realization of conserved residues that represent a protein family is crucial for clearer understanding of biological function as well as for the better recognition of additional members in sequence databases. Functionally important residues are recognized well due to their high degree of conservation in closely related sequences and are annotated in functional motif databases. Structural motifs are central to the integrity of the fold and require careful analysis for their identification. We report the availability of a database of spatially interacting motifs in single protein structures as well as those among distantly related protein structures that belong to a superfamily. Spatial interactions amongst conserved motifs are automatically measured using sequence similarity scores and distance calculations. Interactions between pairs of conserved motifs are described in the form of pseudoenergies. iMOTdb database provides information for 854,488 motifs corresponding to 60,849 protein structural domains and 22,648 protein structural entries. PMID- 16381867 TI - E-MSD: improving data deposition and structure quality. AB - The Macromolecular Structure Database (MSD) (http://www.ebi.ac.uk/msd/) [H. Boutselakis, D. Dimitropoulos, J. Fillon, A. Golovin, K. Henrick, A. Hussain, J. Ionides, M. John, P. A. Keller, E. Krissinel et al. (2003) E-MSD: the European Bioinformatics Institute Macromolecular Structure Database. Nucleic Acids Res., 31, 458-462.] group is one of the three partners in the worldwide Protein DataBank (wwPDB), the consortium entrusted with the collation, maintenance and distribution of the global repository of macromolecular structure data [H. Berman, K. Henrick and H. Nakamura (2003) Announcing the worldwide Protein Data Bank. Nature Struct. Biol., 10, 980.]. Since its inception, the MSD group has worked with partners around the world to improve the quality of PDB data, through a clean up programme that addresses inconsistencies and inaccuracies in the legacy archive. The improvements in data quality in the legacy archive have been achieved largely through the creation of a unified data archive, in the form of a relational database that stores all of the data in the wwPDB. The three partners are working towards improving the tools and methods for the deposition of new data by the community at large. The implementation of the MSD database, together with the parallel development of improved tools and methodologies for data harvesting, validation and archival, has lead to significant improvements in the quality of data that enters the archive. Through this and related projects in the NMR and EM realms the MSD continues to improve the quality of publicly available structural data. PMID- 16381868 TI - The imprinted gene and parent-of-origin effect database now includes parental origin of de novo mutations. AB - The imprinted gene and parent-of-origin effect database (www.otago.ac.nz/IGC) consists of two sections. One section catalogues the current literature on imprinted genes in humans and animals. The second, and new, section catalogues current reports of parental origin of de novo mutations in humans alone. The addition of a catalogue of de novo mutations that show a parent-of-origin effect expands the scope of the database and provides a useful tool for examining parental origin trends for different types of spontaneous mutations. This new section includes >1700 mutations, found in 59 different disorders. The 85 imprinted genes are described in 152 entries from several mammalian species. In addition, >300 other entries describe a range of reported parent-of-origin effects in animals. PMID- 16381869 TI - MODBASE: a database of annotated comparative protein structure models and associated resources. AB - MODBASE (http://salilab.org/modbase) is a database of annotated comparative protein structure models for all available protein sequences that can be matched to at least one known protein structure. The models are calculated by MODPIPE, an automated modeling pipeline that relies on MODELLER for fold assignment, sequence structure alignment, model building and model assessment (http:/salilab.org/modeller). MODBASE is updated regularly to reflect the growth in protein sequence and structure databases, and improvements in the software for calculating the models. MODBASE currently contains 3 094 524 reliable models for domains in 1 094 750 out of 1 817 889 unique protein sequences in the UniProt database (July 5, 2005); only models based on statistically significant alignments and models assessed to have the correct fold despite insignificant alignments are included. MODBASE also allows users to generate comparative models for proteins of interest with the automated modeling server MODWEB (http://salilab.org/modweb). Our other resources integrated with MODBASE include comprehensive databases of multiple protein structure alignments (DBAli, http://salilab.org/dbali), structurally defined ligand binding sites and structurally defined binary domain interfaces (PIBASE, http://salilab.org/pibase) as well as predictions of ligand binding sites, interactions between yeast proteins, and functional consequences of human nsSNPs (LS-SNP, http://salilab.org/LS-SNP). PMID- 16381870 TI - The Database of Macromolecular Motions: new features added at the decade mark. AB - The database of molecular motions, MolMovDB (http://molmovdb.org), has been in existence for the past decade. It classifies macromolecular motions and provides tools to interpolate between two conformations (the Morph Server) and predict possible motions in a single structure. In 2005, we expanded the services offered on MolMovDB. In particular, we further developed the Morph Server to produce improved interpolations between two submitted structures. We added support for multiple chains to the original adiabatic mapping interpolation, allowing the analysis of subunit motions. We also added the option of using FRODA interpolation, which allows for more complex pathways, potentially overcoming steric barriers. We added an interface to a hinge prediction service, which acts on single structures and predicts likely residue points for flexibility. We developed tools to relate such points of flexibility in a structure to particular key residue positions, i.e. active sites or highly conserved positions. Lastly, we began relating our motion classification scheme to function using descriptions from the Gene Ontology Consortium. PMID- 16381871 TI - The Molecular Biology Database Collection: 2006 update. AB - The NAR Molecular Biology Database Collection is a public online resource that contains links to all databases described in this issue of Nucleic Acids Research. In addition, this collection lists databases that have been featured in previous issues of NAR, as well as selected other databases that are freely available to the public and may be useful to the molecular biologist. The 2006 update includes 858 databases, 139 more than the previous one. The databases come with brief summaries, many of which have been updated recently. Each database is assigned a stable accession number that does not change if the database moves to a new location and its URL, authors' names or the contact person address are updated. The complete database list and summaries are available online at the Nucleic Acids Research website http://nar.oxfordjournals.org/. PMID- 16381872 TI - The RCSB PDB information portal for structural genomics. AB - The RCSB Protein Data Bank (PDB) offers online tools, summary reports and target information related to the worldwide structural genomics initiatives from its portal at http://sg.pdb.org. There are currently three components to this site: Structural Genomics Initiatives contains information and links on each structural genomics site, including progress reports, target lists, target status, targets in the PDB and level of sequence redundancy; Targets provides combined target information, protocols and other data associated with protein structure determination; and Structures offers an assessment of the progress of structural genomics based on the functional coverage of the human genome by PDB structures, structural genomics targets and homology models. Functional coverage can be examined according to enzyme classification, gene ontology (biological process, cell component and molecular function) and disease. PMID- 16381873 TI - The PMDB Protein Model Database. AB - The Protein Model Database (PMDB) is a public resource aimed at storing manually built 3D models of proteins. The database is designed to provide access to models published in the scientific literature, together with validating experimental data. It is a relational database and it currently contains >74,000 models for approximately 240 proteins. The system is accessible at http://www.caspur.it/PMDB and allows predictors to submit models along with related supporting evidence and users to download them through a simple and intuitive interface. Users can navigate in the database and retrieve models referring to the same target protein or to different regions of the same protein. Each model is assigned a unique identifier that allows interested users to directly access the data. PMID- 16381874 TI - SCOPPI: a structural classification of protein-protein interfaces. AB - SCOPPI, the structural classification of protein-protein interfaces, is a comprehensive database that classifies and annotates domain interactions derived from all known protein structures. SCOPPI applies SCOP domain definitions and a distance criterion to determine inter-domain interfaces. Using a novel method based on multiple sequence and structural alignments of SCOP families, SCOPPI presents a comprehensive geometrical classification of domain interfaces. Various interface characteristics such as number, type and position of interacting amino acids, conservation, interface size, and permanent or transient nature of the interaction are further provided. Proteins in SCOPPI are annotated with Gene Ontology terms, and the ontology can be used to quickly browse SCOPPI. Screenshots are available for every interface and its participating domains. Here, we describe contents and features of the web-based user interface as well as the underlying methods used to generate SCOPPI's data. In addition, we present a number of examples where SCOPPI becomes a useful tool to analyze viral mimicry of human interface binding sites, gene fusion events, conservation of interface residues and diversity of interface localizations. SCOPPI is available at http://www.scoppi.org. PMID- 16381875 TI - The SWISS-MODEL Repository: new features and functionalities. AB - The SWISS-MODEL Repository is a database of annotated 3D protein structure models generated by the SWISS-MODEL homology-modelling pipeline. As of September 2005, the repository contained 675,000 models for 604,000 different protein sequences of the UniProt database. Regular updates ensure that the content of the repository reflects the current state of sequence and structure databases, integrating new or modified target sequences, and making use of new template structures. Each Repository entry consists of one or more 3D models accompanied by detailed information about the target protein and the model building process: functional annotation, a detailed template selection log, target-template alignment, summary of the model building and model quality assessment. The SWISS MODEL Repository is freely accessible at http://swissmodel.expasy.org/repository/. PMID- 16381876 TI - The HUGO Gene Nomenclature Database, 2006 updates. AB - The HUGO Gene Nomenclature Committee (HGNC) aims to give every human gene a unique and ideally meaningful name and symbol. The HGNC database, previously known as Genew, contains over 22,000 public records with approved human gene nomenclature and associated information. The database has undergone major improvements throughout the last year, is publicly available for online searching at http://www.gene.ucl.ac.uk/cgi-bin/nomenclature/searchgenes.pl and has a new custom downloads interface at http://www.gene.ucl.ac.uk/cgi bin/nomenclature/gdlw.pl. PMID- 16381877 TI - ISfinder: the reference centre for bacterial insertion sequences. AB - ISfinder (www-is.biotoul.fr) is a dedicated database for bacterial insertion sequences (ISs). It has superseded the Stanford reference center. One of its functions is to assign IS names and to provide a focal point for a coherent nomenclature. It is also the repository for ISs. Each new IS is indexed together with information such as its DNA sequence and open reading frames or potential coding sequences, the sequence of the ends of the element and target sites, its origin and distribution together with a bibliography where available. Another objective is to continuously monitor ISs to provide updated comprehensive groupings or families and to provide some insight into their phylogenies. The site also contains extensive background information on ISs and transposons in general. Online tools are gradually being added. At present an online Blast facility against the entire bank is available. But additional features will include alignment capability, PsiBLAST and HMM profiles. ISfinder also includes a section on bacterial genomes and is involved in annotating the IS content of these genomes. Finally, this database is currently recommended by several microbiology journals for registration of new IS elements before their publication. PMID- 16381878 TI - The Gene Ontology (GO) project in 2006. AB - The Gene Ontology (GO) project (http://www.geneontology.org) develops and uses a set of structured, controlled vocabularies for community use in annotating genes, gene products and sequences (also see http://song.sourceforge.net/). The GO Consortium continues to improve to the vocabulary content, reflecting the impact of several novel mechanisms of incorporating community input. A growing number of model organism databases and genome annotation groups contribute annotation sets using GO terms to GO's public repository. Updates to the AmiGO browser have improved access to contributed genome annotations. As the GO project continues to grow, the use of the GO vocabularies is becoming more varied as well as more widespread. The GO project provides an ontological annotation system that enables biologists to infer knowledge from large amounts of data. PMID- 16381879 TI - PANDIT: an evolution-centric database of protein and associated nucleotide domains with inferred trees. AB - PANDIT is a database of homologous sequence alignments accompanied by estimates of their corresponding phylogenetic trees. It provides a valuable resource to those studying phylogenetic methodology and the evolution of coding-DNA and protein sequences. Currently in version 17.0, PANDIT comprises 7738 families of homologous protein domains; for each family, DNA and corresponding amino acid sequence multiple alignments are available together with high quality phylogenetic tree estimates. Recent improvements include expanded methods for phylogenetic tree inference, assessment of alignment quality and a redesigned web interface, available at the URL http://www.ebi.ac.uk/goldman-srv/pandit. PMID- 16381880 TI - The Genomes On Line Database (GOLD) v.2: a monitor of genome projects worldwide. AB - The Genomes On Line Database (GOLD) is a web resource for comprehensive access to information regarding complete and ongoing genome sequencing projects worldwide. The database currently incorporates information on over 1500 sequencing projects, of which 294 have been completed and the data deposited in the public databases. GOLD v.2 has been expanded to provide information related to organism properties such as phenotype, ecotype and disease. Furthermore, project relevance and availability information is now included. GOLD is available at http://www.genomesonline.org. It is also mirrored at the Institute of Molecular Biology and Biotechnology, Crete, Greece at http://gold.imbb.forth.gr/ PMID- 16381881 TI - xBASE, a collection of online databases for bacterial comparative genomics. AB - The schema of the previously described Escherischia coli database coliBASE has been applied to a number of other bacterial taxa, under the collective name xBASE. The new databases include CampyDB for Campylobacter, Helicobacter and Wolinella; PseudoDB for pseudomonads; ClostriDB for clostridia; RhizoDB for Rhizobium and Sinorhizobium; and MycoDB, for Mycobacterium, Streptomyces and related organisms. The databases provide user friendly access to annotation and genome comparisons through a web-based graphical interface. Newly developed features include whole genome displays, 'painting' of genes according to properties such as GC content, a pattern search system to identify conserved motifs and batch BLAST searching of every protein encoded by a region. Examples of how the databases have been, and continue to be, used to generate hypotheses for subsequent laboratory investigation are presented. xBASE is available online at http://xbase.bham.ac.uk. PMID- 16381882 TI - ICDS database: interrupted CoDing sequences in prokaryotic genomes. AB - Unrecognized frameshifts, in-frame stop codons and sequencing errors lead to Interrupted CoDing Sequence (ICDS) that can seriously affect all subsequent steps of functional characterization, from in silico analysis to high-throughput proteomic projects. Here, we describe the Interrupted CoDing Sequence database containing ICDS detected by a similarity-based approach in 80 complete prokaryotic genomes. ICDS can be retrieved by species browsing or similarity searches via a web interface (http://www-bio3d-igbmc.u-strasbg.fr/ICDS/). The definition of each interrupted gene is provided as well as the ICDS genomic localization with the surrounding sequence. Furthermore, to facilitate the experimental characterization of ICDS, we propose optimized primers for re sequencing purposes. The database will be regularly updated with additional data from ongoing sequenced genomes. Our strategy has been validated by three independent tests: (i) ICDS prediction on a benchmark of artificially created frameshifts, (ii) comparison of predicted ICDS and results obtained from the comparison of the two genomic sequences of Bacillus licheniformis strain ATCC 14580 and (iii) re-sequencing of 25 predicted ICDS of the recently sequenced genome of Mycobacterium smegmatis. This allows us to estimate the specificity and sensitivity (95 and 82%, respectively) of our program and the efficiency of primer determination. PMID- 16381883 TI - The integrated microbial genomes (IMG) system. AB - The integrated microbial genomes (IMG) system is a new data management and analysis platform for microbial genomes provided by the Joint Genome Institute (JGI). IMG contains both draft and complete JGI genomes integrated with other publicly available microbial genomes of all three domains of life. IMG provides tools and viewers for analyzing genomes, genes and functions, individually or in a comparative context. IMG allows users to focus their analysis on subsets of genes and genomes of interest and to save the results of their analysis. IMG is available at http://img.jgi.doe.gov. PMID- 16381884 TI - INVHOGEN: a database of homologous invertebrate genes. AB - Classification of proteins into families of homologous sequences constitutes the basis of functional analysis or of evolutionary studies. Here we present INVertebrate HOmologous GENes (INVHOGEN), a database combining the available invertebrate protein genes from UniProt (consisting of Swiss-Prot and TrEMBL) into gene families. For each family INVHOGEN provides a multiple protein alignment, a maximum likelihood based phylogenetic tree and taxonomic information about the sequences. It is possible to download the corresponding GenBank flatfiles, the alignment and the tree in Newick format. Sequences and related information have been structured in an ACNUC database under a client/server architecture. Thus, complex selections can be performed. An external graphical tool (FamFetch) allows access to the data to evaluate homology relationships between genes and distinguish orthologous from paralogous sequences. Thus, INVHOGEN complements the well-known HOVERGEN database. The databank is available at http://www.bi.uni-duesseldorf.de/~invhogen/invhogen.html. PMID- 16381885 TI - From genomics to chemical genomics: new developments in KEGG. AB - The increasing amount of genomic and molecular information is the basis for understanding higher-order biological systems, such as the cell and the organism, and their interactions with the environment, as well as for medical, industrial and other practical applications. The KEGG resource (http://www.genome.jp/kegg/) provides a reference knowledge base for linking genomes to biological systems, categorized as building blocks in the genomic space (KEGG GENES) and the chemical space (KEGG LIGAND), and wiring diagrams of interaction networks and reaction networks (KEGG PATHWAY). A fourth component, KEGG BRITE, has been formally added to the KEGG suite of databases. This reflects our attempt to computerize functional interpretations as part of the pathway reconstruction process based on the hierarchically structured knowledge about the genomic, chemical and network spaces. In accordance with the new chemical genomics initiatives, the scope of KEGG LIGAND has been significantly expanded to cover both endogenous and exogenous molecules. Specifically, RPAIR contains curated chemical structure transformation patterns extracted from known enzymatic reactions, which would enable analysis of genome-environment interactions, such as the prediction of new reactions and new enzyme genes that would degrade new environmental compounds. Additionally, drug information is now stored separately and linked to new KEGG DRUG structure maps. PMID- 16381886 TI - ODB: a database of operons accumulating known operons across multiple genomes. AB - Operon structures play an important role in co-regulation in prokaryotes. Although over 200 complete genome sequences are now available, databases providing genome-wide operon information have been limited to certain specific genomes. Thus, we have developed an ODB (Operon DataBase), which provides a data retrieval system of known operons among the many complete genomes. Additionally, putative operons that are conserved in terms of known operons are also provided. The current version of our database contains about 2000 known operon information in more than 50 genomes and about 13 000 putative operons in more than 200 genomes. This system integrates four types of associations: genome context, gene co-expression obtained from microarray data, functional links in biological pathways and the conservation of gene order across the genomes. These associations are indicators of the genes that organize an operon, and the combination of these indicators allows us to predict more reliable operons. Furthermore, our system validates these predictions using known operon information obtained from the literature. This database integrates known literature-based information and genomic data. In addition, it provides an operon prediction tool, which make the system useful for both bioinformatics researchers and experimental biologists. Our database is accessible at http://odb.kuicr.kyoto u.ac.jp/. PMID- 16381887 TI - OrthoMCL-DB: querying a comprehensive multi-species collection of ortholog groups. AB - The OrthoMCL database (http://orthomcl.cbil.upenn.edu) houses ortholog group predictions for 55 species, including 16 bacterial and 4 archaeal genomes representing phylogenetically diverse lineages, and most currently available complete eukaryotic genomes: 24 unikonts (12 animals, 9 fungi, microsporidium, Dictyostelium, Entamoeba), 4 plants/algae and 7 apicomplexan parasites. OrthoMCL software was used to cluster proteins based on sequence similarity, using an all against-all BLAST search of each species' proteome, followed by normalization of inter-species differences, and Markov clustering. A total of 511,797 proteins (81.6% of the total dataset) were clustered into 70,388 ortholog groups. The ortholog database may be queried based on protein or group accession numbers, keyword descriptions or BLAST similarity. Ortholog groups exhibiting specific phyletic patterns may also be identified, using either a graphical interface or a text-based Phyletic Pattern Expression grammar. Information for ortholog groups includes the phyletic profile, the list of member proteins and a multiple sequence alignment, a statistical summary and graphical view of similarities, and a graphical representation of domain architecture. OrthoMCL software, the entire FASTA dataset employed and clustering results are available for download. OrthoMCL-DB provides a centralized warehouse for orthology prediction among multiple species, and will be updated and expanded as additional genome sequence data become available. PMID- 16381888 TI - PUMA2--grid-based high-throughput analysis of genomes and metabolic pathways. AB - The PUMA2 system (available at http://compbio.mcs.anl.gov/puma2) is an interactive, integrated bioinformatics environment for high-throughput genetic sequence analysis and metabolic reconstructions from sequence data. PUMA2 provides a framework for comparative and evolutionary analysis of genomic data and metabolic networks in the context of taxonomic and phenotypic information. Grid infrastructure is used to perform computationally intensive tasks. PUMA2 currently contains precomputed analysis of 213 prokaryotic, 22 eukaryotic, 650 mitochondrial and 1493 viral genomes and automated metabolic reconstructions for >200 organisms. Genomic data is annotated with information integrated from >20 sequence, structural and metabolic databases and ontologies. PUMA2 supports both automated and interactive expert-driven annotation of genomes, using a variety of publicly available bioinformatics tools. It also contains a suite of unique PUMA2 tools for automated assignment of gene function, evolutionary analysis of protein families and comparative analysis of metabolic pathways. PUMA2 allows users to submit batch sequence data for automated functional analysis and construction of metabolic models. The results of these analyses are made available to the users in the PUMA2 environment for further interactive sequence analysis and annotation. PMID- 16381889 TI - Transterm--extended search facilities and improved integration with other databases. AB - Transterm has now been publicly available for >10 years. Major changes have been made since its last description in this database issue in 2002. The current database provides data for key regions of mRNA sequences, a curated database of mRNA motifs and tools to allow users to investigate their own motifs or mRNA sequences. The key mRNA regions database is derived computationally from Genbank. It contains 3' and 5' flanking regions, the initiation and termination signal context and coding sequence for annotated CDS features from Genbank and RefSeq. The database is non-redundant, enabling summary files and statistics to be prepared for each species. Advances include providing extended search facilities, the database may now be searched by BLAST in addition to regular expressions (patterns) allowing users to search for motifs such as known miRNA sequences, and the inclusion of RefSeq data. The database contains >40 motifs or structural patterns important for translational control. In this release, patterns from UTRsite and Rfam are also incorporated with cross-referencing. Users may search their sequence data with Transterm or user-defined patterns. The system is accessible at http://uther.otago.ac.nz/Transterm.html. PMID- 16381890 TI - SIDDBASE: a database containing the stress-induced DNA duplex destabilization (SIDD) profiles of complete microbial genomes. AB - Prokaryotic genomic DNA is generally negatively supercoiled in vivo. Many regulatory processes, including the initiation of transcription, are known to depend on the superhelical state of the DNA substrate. The stresses induced within DNA by negative superhelicity can destabilize the DNA duplex at specific sites. Various experiments have either shown or suggested that stress-induced DNA duplex destabilization (SIDD) is involved in specific regulatory mechanisms governing a variety of biological processes. We have developed methods to evaluate the SIDD properties of DNA sequences, including complete chromosomes. This analysis predicts the locations where the duplex becomes destabilized under superhelical stress. Previous studies have shown that the SIDD-susceptible sites predicted in this way occur at rates much higher than expected at random in transcriptional regulatory regions, and much lower than expected in coding regions. Analysis of the SIDD profiles of 42 bacterial genomes chosen for their diversity confirms this pattern. Predictions of SIDD sites have been used to identify potential genomic regulatory regions, and suggest both possible regulatory mechanisms involving stress-induced destabilization and experimental tests of these mechanisms. Here we describe the SIDDBASE database which enables users to retrieve and visualize the results of SIDD analyses of completely sequenced prokaryotic and archaeal genomes, together with their annotations. SIDDBASE is available at www.gc.ucdavis.edu/benham/siddbase. PMID- 16381891 TI - VMD: a community annotation database for oomycetes and microbial genomes. AB - The VBI Microbial Database (VMD) is a database system designed to host a range of microbial genome sequences. At present, the database contains genome sequence and annotation data of two plant pathogens Phytophthora sojae and Phytophthora ramorum. With the completion of the draft genome sequences of these pathogens in collaboration with the DOE Joint Genome Institute (JGI), we have created this resource to make the sequences publicly available. The genome sequences (95 MB for P.sojae and 65 MB for P.ramorum) were annotated with approximately 19,000 and approximately 16,000 gene models, respectively. We used two different statistical methods to validate these gene models, Fickett's and a log-likelihood method. Functional annotation of the gene models is based on results from BlastX and InterProScan screens. From the InterProScan results, we could assign putative functions to 17,694 genes in P.sojae and 14,700 genes in P.ramorum. We created an easy-to-use genome browser to view the genome sequence data, which opens to detailed annotation pages for each gene model. A community annotation interface is available for registered community members to add or edit annotations. There are approximately 1600 gene models for P.sojae and approximately 700 models for P.ramorum that have already been manually curated. A toolkit is provided as an additional resource for users to perform a variety of sequence analysis jobs. The database is publicly available at http://phytophthora.vbi.vt.edu/. PMID- 16381892 TI - DPVweb: a comprehensive database of plant and fungal virus genes and genomes. AB - DPVweb (http://www.dpvweb.net/) provides a central source of information about viruses, viroids and satellites of plants, fungi and protozoa. Comprehensive taxonomic information, including brief descriptions of each family and genus, and classified lists of virus sequences are provided. The database also holds detailed, curated, information for all sequences of viruses, viroids and satellites of plants, fungi and protozoa that are complete or that contain at least one complete gene (currently, n approximately 9000). For comparative purposes, it also contains a single representative sequence of all other fully sequenced virus species with an RNA or single-stranded DNA genome. The start and end positions of each feature (gene, non-translated region and the like) have been recorded and checked for accuracy. As far as possible, nomenclature for genes and proteins are standardized within genera and families. Sequences of features (either as DNA or amino acid sequences) can be directly downloaded from the website in FASTA format. The sequence information can also be accessed via client software for PC computers (freely downloadable from the website) that enable users to make an easy selection of sequences and features of a chosen virus for further analyses. PMID- 16381893 TI - VIPERdb: a relational database for structural virology. AB - VIPERdb (http://viperdb.scripps.edu) is a database for icosahedral virus capsid structures. Our aim is to provide a comprehensive resource specific to the needs of the structural virology community, with an emphasis on the description and comparison of derived data from structural and energetic analyses of capsids. A relational database implementation based on a schema for macromolecular structure makes the data highly accessible to the user, allowing detailed queries at the atomic level. Together with curation practices that maintain data uniformity, this will facilitate structural bioinformatics studies of virus capsids. User friendly search, visualization and educational tools on the website allow both structural and derived data to be examined easily and extensively. Links to relevant literature, sequence and taxonomy databases are provided for each entry. PMID- 16381894 TI - Megx.net--database resources for marine ecological genomics. AB - Marine microbial genomics and metagenomics is an emerging field in environmental research. Since the completion of the first marine bacterial genome in 2003, the number of fully sequenced marine bacteria has grown rapidly. Concurrently, marine metagenomics studies are performed on a regular basis, and the resulting number of sequences is growing exponentially. To address environmentally relevant questions like organismal adaptations to oceanic provinces and regional differences in the microbial cycling of nutrients, it is necessary to couple sequence data with geographical information and supplement them with contextual information like physical, chemical and biological data. Therefore, new specialized databases are needed to organize and standardize data storage as well as centralize data access and interpretation. We introduce Megx.net, a set of databases and tools that handle genomic and metagenomic sequences in their environmental contexts. Megx.net includes (i) a geographic information system to systematically store and analyse marine genomic and metagenomic data in conjunction with contextual information; (ii) an environmental genome browser with fast search functionalities; (iii) a database with precomputed analyses for selected complete genomes; and (iv) a database and tool to classify metagenomic fragments based on oligonucleotide signatures. These integrative databases and webserver will help researchers to generate a better understanding of the functioning of marine ecosystems. All resources are freely accessible at http://www.megx.net. PMID- 16381895 TI - RegulonDB (version 5.0): Escherichia coli K-12 transcriptional regulatory network, operon organization, and growth conditions. AB - RegulonDB is the internationally recognized reference database of Escherichia coli K-12 offering curated knowledge of the regulatory network and operon organization. It is currently the largest electronically-encoded database of the regulatory network of any free-living organism. We present here the recently launched RegulonDB version 5.0 radically different in content, interface design and capabilities. Continuous curation of original scientific literature provides the evidence behind every single object and feature. This knowledge is complemented with comprehensive computational predictions across the complete genome. Literature-based and predicted data are clearly distinguished in the database. Starting with this version, RegulonDB public releases are synchronized with those of EcoCyc since our curation supports both databases. The complex biology of regulation is simplified in a navigation scheme based on three major streams: genes, operons and regulons. Regulatory knowledge is directly available in every navigation step. Displays combine graphic and textual information and are organized allowing different levels of detail and biological context. This knowledge is the backbone of an integrated system for the graphic display of the network, graphic and tabular microarray comparisons with curated and predicted objects, as well as predictions across bacterial genomes, and predicted networks of functionally related gene products. Access RegulonDB at http://regulondb.ccg.unam.mx. PMID- 16381896 TI - ShiBASE: an integrated database for comparative genomics of Shigella. AB - Among the major enteric bacterial pathogens, Shigella is found to display extreme genome diversity and dynamics, which imposes a challenge in comparative genomic studies. To facilitate further studies in this area, we have constructed an integrated online database, ShiBASE (http://www.mgc.ac.cn/ShiBASE/),which contains Shigella genomic sequences of four species and additional comparative genomic hybridization (CGH) data of 43 serotypes. ShiBASE offers online comparative analysis on DNA sequences, gene orders, metabolic pathways and virulence factors. In addition, ShiBASE has a newly developed online comparative visualization service, Shi-align, which enables the alignment of any query sequence with the reference genome sequences. PMID- 16381897 TI - LEGER: knowledge database and visualization tool for comparative genomics of pathogenic and non-pathogenic Listeria species. AB - Listeria species are ubiquitous in the environment and often contaminate foods because they grow under conditions used for food preservation. Listeria monocytogenes, the human and animal pathogen, causes Listeriosis, an infection with a high mortality rate in risk groups such as immune-compromised individuals. Furthermore, L.monocytogenes is a model organism for the study of intracellular bacterial pathogens. The publication of its genome sequence and that of the non pathogenic species Listeria innocua initiated numerous comparative studies and efforts to sequence all species comprising the genus. The Proteome database LEGER (http://leger2.gbf.de/cgi-bin/expLeger.pl) was developed to support functional genome analyses by combining information obtained by applying bioinformatics methods and from public databases to improve the original annotations. LEGER offers three unique key features: (i) it is the first comprehensive information system focusing on the functional assignment of genes and proteins; (ii) integrated visualization tools, KEGG pathway and Genome Viewer, alleviate the functional exploration of complex data; and (iii) LEGER presents results of systematic post-genome studies, thus facilitating analyses combining computational and experimental results. Moreover, LEGER provides an unpublished membrane proteome analysis of L.innocua and in total visualizes experimentally validated information about the subcellular localizations of 789 different listerial proteins. PMID- 16381898 TI - The UCSC Archaeal Genome Browser. AB - As more archaeal genomes are sequenced, effective research and analysis tools are needed to integrate the diverse information available for any given locus. The feature-rich UCSC Genome Browser, created originally to annotate the human genome, can be applied to any sequenced organism. We have created a UCSC Archaeal Genome Browser, available at http://archaea.ucsc.edu/, currently with 26 archaeal genomes. It displays G/C content, gene and operon annotation from multiple sources, sequence motifs (promoters and Shine-Dalgarno), microarray data, multi genome alignments and protein conservation across phylogenetic and habitat categories. We encourage submission of new experimental and bioinformatic analysis from contributors. The purpose of this tool is to aid biological discovery and facilitate greater collaboration within the archaeal research community. PMID- 16381899 TI - ASAP: a resource for annotating, curating, comparing, and disseminating genomic data. AB - ASAP is a comprehensive web-based system for community genome annotation and analysis. ASAP is being used for a large-scale effort to augment and curate annotations for genomes of enterobacterial pathogens and for additional genome sequences. New tools, such as the genome alignment program Mauve, have been incorporated into ASAP in order to improve display and analysis of related genomes. Recent improvements to the database and challenges for future development of the system are discussed. ASAP is available on the web at https://asap.ahabs.wisc.edu/asap/logon.php. PMID- 16381900 TI - Human protein reference database--2006 update. AB - Human Protein Reference Database (HPRD) (http://www.hprd.org) was developed to serve as a comprehensive collection of protein features, post-translational modifications (PTMs) and protein-protein interactions. Since the original report, this database has increased to >20 000 proteins entries and has become the largest database for literature-derived protein-protein interactions (>30 000) and PTMs (>8000) for human proteins. We have also introduced several new features in HPRD including: (i) protein isoforms, (ii) enhanced search options, (iii) linking of pathway annotations and (iv) integration of a novel browser, GenProt Viewer (http://www.genprot.org), developed by us that allows integration of genomic and proteomic information. With the continued support and active participation by the biomedical community, we expect HPRD to become a unique source of curated information for the human proteome and spur biomedical discoveries based on integration of genomic, transcriptomic and proteomic data. PMID- 16381901 TI - The LIFEdb database in 2006. AB - LIFEdb (http://www.LIFEdb.de) integrates data from large-scale functional genomics assays and manual cDNA annotation with bioinformatics gene expression and protein analysis. New features of LIFEdb include (i) an updated user interface with enhanced query capabilities, (ii) a configurable output table and the option to download search results in XML, (iii) the integration of data from cell-based screening assays addressing the influence of protein-overexpression on cell proliferation and (iv) the display of the relative expression ('Electronic Northern') of the genes under investigation using curated gene expression ontology information. LIFEdb enables researchers to systematically select and characterize genes and proteins of interest, and presents data and information via its user-friendly web-based interface. PMID- 16381902 TI - CryptoDB: a Cryptosporidium bioinformatics resource update. AB - The database, CryptoDB (http://CryptoDB.org), is a community bioinformatics resource for the AIDS-related apicomplexan-parasite, Cryptosporidium. CryptoDB integrates whole genome sequence and annotation with expressed sequence tag and genome survey sequence data and provides supplemental bioinformatics analyses and data-mining tools. A simple, yet comprehensive web interface is available for mining and visualizing the data. CryptoDB is allied with the databases PlasmoDB and ToxoDB via ApiDB, an NIH/NIAID-fundedBioinformatics Resource Center. Recent updates to CryptoDB include the deposition of annotated genome sequences for Cryptosporidium parvum and Cryptosporidium hominis, migration to a relational database (GUS), a new query and visualization interface and the introduction of Web services. PMID- 16381903 TI - dictyBase, the model organism database for Dictyostelium discoideum. AB - dictyBase (http://dictybase.org) is the model organism database (MOD) for the social amoeba Dictyostelium discoideum. The unique biology and phylogenetic position of Dictyostelium offer a great opportunity to gain knowledge of processes not characterized in other organisms. The recent completion of the 34 MB genome sequence, together with the sizable scientific literature using Dictyostelium as a research organism, provided the necessary tools to create a well-annotated genome. dictyBase has leveraged software developed by the Saccharomyces Genome Database and the Generic Model Organism Database project. This has reduced the time required to develop a full-featured MOD and greatly facilitated our ability to focus on annotation and providing new functionality. We hope that manual curation of the Dictyostelium genome will facilitate the annotation of other genomes. PMID- 16381904 TI - TcruziDB: an integrated, post-genomics community resource for Trypanosoma cruzi. AB - TcruziDB (http://TcruziDB.org) is an integrated post-genomics database for the parasitic organism, Trypanosoma cruzi, the causative agent of Chagas' disease. TcruziDB was established in 2003 as a flat-file database with tools for mining the unannotated sequence reads and preliminary contig assemblies emerging from the Tri-Tryp genome consortium (TIGR/SBRI/Karolinska). Today, TcruziDB houses the recently published assembled genomic contigs and annotation provided by the genome consortium in a relational database supported by the Genomics Unified Schema (GUS) architecture. The combination of an annotated genome and a relational architecture has facilitated the integration of genomic data with expression data (proteomic and EST) and permitted the construction of automated analysis pipelines. TcruziDB has accepted, and will continue to accept the deposition of genomic and functional genomic datasets contributed by the research community. PMID- 16381905 TI - Genolevures complete genomes provide data and tools for comparative genomics of hemiascomycetous yeasts. AB - The Genolevures online database (http://cbi.labri.fr/Genolevures/) provides tools and data relative to 4 complete and 10 partial genome sequences determined and manually annotated by the Genolevures Consortium, to facilitate comparative genomic studies of hemiascomycetous yeasts. With their relatively small and compact genomes, yeasts offer a unique opportunity for exploring eukaryotic genome evolution. The new version of the Genolevures database provides truly complete (subtelomere to subtelomere) chromosome sequences, 25 000 protein-coding and tRNA genes, and in silico analyses for each gene element. A new feature of the database is a novel collection of conserved multi-species protein families and their mapping to metabolic pathways, coupled with an advanced search feature. Data are presented with a focus on relations between genes and genomes: conservation of genes and gene families, speciation, chromosomal reorganization and synteny. The Genolevures site includes an area for specific studies by members of its international community. PMID- 16381906 TI - MPact: the MIPS protein interaction resource on yeast. AB - In recent years, the Munich Information Center for Protein Sequences (MIPS) yeast protein-protein interaction (PPI) dataset has been used in numerous analyses of protein networks and has been called a gold standard because of its quality and comprehensiveness [H. Yu, N. M. Luscombe, H. X. Lu, X. Zhu, Y. Xia, J. D. Han, N. Bertin, S. Chung, M. Vidal and M. Gerstein (2004) Genome Res., 14, 1107-1118]. MPact and the yeast protein localization catalog provide information related to the proximity of proteins in yeast. Beside the integration of high-throughput data, information about experimental evidence for PPIs in the literature was compiled by experts adding up to 4300 distinct PPIs connecting 1500 proteins in yeast. As the interaction data is a complementary part of CYGD, interactive mapping of data on other integrated data types such as the functional classification catalog [A. Ruepp, A. Zollner, D. Maier, K. Albermann, J. Hani, M. Mokrejs, I. Tetko, U. Guldener, G. Mannhaupt, M. Munsterkotter and H. W. Mewes (2004) Nucleic Acids Res., 32, 5539-5545] is possible. A survey of signaling proteins and comparison with pathway data from KEGG demonstrates that based on these manually annotated data only an extensive overview of the complexity of this functional network can be obtained in yeast. The implementation of a web based PPI-analysis tool allows analysis and visualization of protein interaction networks and facilitates integration of our curated data with high-throughput datasets. The complete dataset as well as user-defined sub-networks can be retrieved easily in the standardized PSI-MI format. The resource can be accessed through http://mips.gsf.de/genre/proj/mpact. PMID- 16381907 TI - Genome Snapshot: a new resource at the Saccharomyces Genome Database (SGD) presenting an overview of the Saccharomyces cerevisiae genome. AB - Sequencing and annotation of the entire Saccharomyces cerevisiae genome has made it possible to gain a genome-wide perspective on yeast genes and gene products. To make this information available on an ongoing basis, the Saccharomyces Genome Database (SGD) (http://www.yeastgenome.org/) has created the Genome Snapshot (http://db.yeastgenome.org/cgi-bin/genomeSnapShot.pl). The Genome Snapshot summarizes the current state of knowledge about the genes and chromosomal features of S.cerevisiae. The information is organized into two categories: (i) number of each type of chromosomal feature annotated in the genome and (ii) number and distribution of genes annotated to Gene Ontology terms. Detailed lists are accessible through SGD's Advanced Search tool (http://db.yeastgenome.org/cgi bin/search/featureSearch), and all the data presented on this page are available from the SGD ftp site (ftp://ftp.yeastgenome.org/yeast/). PMID- 16381909 TI - Visualizing syntenic relationships among the hemiascomycetes with the Yeast Gene Order Browser. AB - The Yeast Gene Order Browser (YGOB) is an online tool designed to facilitate the comparative genomic visualization and appraisal of synteny within and between the genomes of seven hemiascomycete yeast species. Three of these genomes are polyploid, and hence contain intra-genomic syntenic regions, the correct assembly of which is a particular success of YGOB. Designed to accurately assemble, display and score gene order relationships, YGOB is both an interactive tool for browsing genomic data, and a software engine now being used for evolutionary analyses on a whole-genome scale. Underlying the online interface is the YGOB database, which consists of homology assignments across the species, extensively curated based on sequence similarity and novelly, an appraisal of genomic context (synteny) in multiple genomes. Currently the YGOB database incorporates genome data from Saccharomyces cerevisiae, Candida glabrata, Saccharomyces castellii, Ashbya gossypii, Kluyveromyces lactis, Kluyveromyces waltii and Saccharomyces kluyveri, but the system is scaleable to accommodate additional genomes. This paper discusses the usage and utility of version 1.0 of YGOB, which is publicly available at http://wolfe.gen.tcd.ie/ygob. PMID- 16381908 TI - The YEASTRACT database: a tool for the analysis of transcription regulatory associations in Saccharomyces cerevisiae. AB - We present the YEAst Search for Transcriptional Regulators And Consensus Tracking (YEASTRACT; www.yeastract.com) database, a tool for the analysis of transcription regulatory associations in Saccharomyces cerevisiae. This database is a repository of 12 346 regulatory associations between transcription factors and target genes, based on experimental evidence which was spread throughout 861 bibliographic references. It also includes 257 specific DNA-binding sites for more than a hundred characterized transcription factors. Further information about each yeast gene included in the database was obtained from Saccharomyces Genome Database (SGD), Regulatory Sequences Analysis Tools and Gene Ontology (GO) Consortium. Computational tools are also provided to facilitate the exploitation of the gathered data when solving a number of biological questions as exemplified in the Tutorial also available on the system. YEASTRACT allows the identification of documented or potential transcription regulators of a given gene and of documented or potential regulons for each transcription factor. It also renders possible the comparison between DNA motifs, such as those found to be over represented in the promoter regions of co-regulated genes, and the transcription factor-binding sites described in the literature. The system also provides an useful mechanism for grouping a list of genes (for instance a set of genes with similar expression profiles as revealed by microarray analysis) based on their regulatory associations with known transcription factors. PMID- 16381910 TI - FGDB: a comprehensive fungal genome resource on the plant pathogen Fusarium graminearum. AB - The MIPS Fusarium graminearum Genome Database (FGDB) is a comprehensive genome database on one of the most devastating fungal plant pathogens of wheat and barley. FGDB provides information on two gene sets independently derived by automated annotation of the F.graminearum genome sequence. A complete manually revised gene set will be completed within the near future. The initial results of systematic manual correction of gene calls are already part of the current gene set. The database can be accessed to retrieve information from bioinformatics analyses and functional classifications of the proteins. The data are also organized in the well established MIPS catalogs and novel query techniques are available to search the data. The comprehensive set of gene calls was also used for the design of an Affymetrix GeneChip. The resource is accessible on http://mips.gsf.de/genre/proj/fusarium/. PMID- 16381911 TI - PHI-base: a new database for pathogen host interactions. AB - To utilize effectively the growing number of verified genes that mediate an organism's ability to cause disease and/or to trigger host responses, we have developed PHI-base. This is a web-accessible database that currently catalogs 405 experimentally verified pathogenicity, virulence and effector genes from 54 fungal and Oomycete pathogens, of which 176 are from animal pathogens, 227 from plant pathogens and 3 from pathogens with a fungal host. PHI-base is the first on line resource devoted to the identification and presentation of information on fungal and Oomycete pathogenicity genes and their host interactions. As such, PHI base is a valuable resource for the discovery of candidate targets in medically and agronomically important fungal and Oomycete pathogens for intervention with synthetic chemistries and natural products. Each entry in PHI-base is curated by domain experts and supported by strong experimental evidence (gene/transcript disruption experiments) as well as literature references in which the experiments are described. Each gene in PHI-base is presented with its nucleotide and deduced amino acid sequence as well as a detailed description of the predicted protein's function during the host infection process. To facilitate data interoperability, we have annotated genes using controlled vocabularies (Gene Ontology terms, Enzyme Commission Numbers and so on), and provide links to other external data sources (e.g. NCBI taxonomy and EMBL). We welcome new data for inclusion in PHI base, which is freely accessed at www4.rothamsted.bbsrc.ac.uk/phibase/. PMID- 16381912 TI - ASD: a bioinformatics resource on alternative splicing. AB - Alternative splicing is an important regulatory mechanism of mammalian gene expression. The alternative splicing database (ASD) consortium is systematically collecting and annotating data on alternative splicing. We present the continuation and upgrade of the ASD [T. A. Thanaraj, S. Stamm, F. Clark, J. J. Riethoven, V. Le Texier, J. Muilu (2004) Nucleic Acids Res. 32, D64-D69] that consists of computationally and manually generated data. Its largest parts are AltSplice, a value-added database of computationally delineated alternative splicing events. Its data include alternatively spliced introns/exons, events, isoform splicing patterns and isoform peptide sequences. AltSplice data are generated by examining gene-transcript alignments. The data are annotated for various biological features including splicing signals, expression states, (SNP) mediated splicing and cross-species conservation. AEdb forms the manually curated component of ASD. It is a literature-based data set containing sequence and properties of alternatively spliced exons, functional enumeration of observed splicing events, characterization of observed splicing regulatory elements, and a collection of experimentally clarified minigene constructs. ASD includes a workbench, which is an analysis tool that enables users to carry out splicing related analysis such as characterization of introns for various splicing signals, identification of splicing regulatory elements on a given RNA sequence, prediction of putative exons and prediction of putative translation start codons. The different ASD modules are integrated and can be accessed through user friendly interfaces and visualization tools. ASD data has been integrated with Ensembl genome annotation project as a Distributed Annotation System (DAS) resource and can be viewed on Ensembl genome browser. The ASD resource is presented at (http://www.ebi.ac.uk/asd). PMID- 16381913 TI - Phytophthora functional genomics database (PFGD): functional genomics of phytophthora-plant interactions. AB - The Phytophthora Functional Genomics Database (PFGD; http://www.pfgd.org), developed by the National Center for Genome Resources in collaboration with The Ohio State University-Ohio Agricultural Research and Development Center (OSU OARDC), is a publicly accessible information resource for Phytophthora-plant interaction research. PFGD contains transcript, genomic, gene expression and functional assay data for Phytophthora infestans, which causes late blight of potato, and Phytophthora sojae, which affects soybeans. Automated analyses are performed on all sequence data, including consensus sequences derived from clustered and assembled expressed sequence tags. The PFGD search filter interface allows intuitive navigation of transcript and genomic data organized by library and derived queries using modifiers, annotation keywords or sequence names. BLAST services are provided for libraries built from the transcript and genomic sequences. Transcript data visualization tools include Quality Screening, Multiple Sequence Alignment and Features and Annotations viewers. A genomic browser that supports comparative analysis via novel dynamic functional annotation comparisons is also provided. PFGD is integrated with the Solanaceae Genomics Database (SolGD; http://www.solgd.org) to help provide insight into the mechanisms of infection and resistance, specifically as they relate to the genus Phytophthora pathogens and their plant hosts. PMID- 16381914 TI - SGCEdb: a flexible database and web interface integrating experimental results and analysis for structural genomics focusing on Caenorhabditis elegans. AB - The SGCEdb (http://sgce.cbse.uab.edu) database/interface serves the primary purpose of reporting progress of the Structural Genomics of Caenorhabditis elegans project at the University of Alabama at Birmingham. It stores and analyzes results of experiments ranging from solubility screening arrays to individual protein purification and structure solution. External databases and algorithms are referenced and evaluated for target selection in the human, C.elegans and Pneumocystis carinii genomes. The flexible and reusable design permits tracking of standard and custom experiment types in a scientist-defined sequence. The database coordinates efforts between collaborators and is adaptable to a wide range of biological applications. PMID- 16381915 TI - WormBase: better software, richer content. AB - WormBase (http://wormbase.org), the public database for genomics and biology of Caenorhabditis elegans, has been restructured for stronger performance and expanded for richer biological content. Performance was improved by accelerating the loading of central data pages such as the omnibus Gene page, by rationalizing internal data structures and software for greater portability, and by making the Genome Browser highly customizable in how it views and exports genomic subsequences. Arbitrarily complex, user-specified queries are now possible through Textpresso (for all available literature) and through WormMart (for most genomic data). Biological content was enriched by reconciling all available cDNA and expressed sequence tag data with gene predictions, clarifying single nucleotide polymorphism and RNAi sites, and summarizing known functions for most genes studied in this organism. PMID- 16381916 TI - FLIGHT: database and tools for the integration and cross-correlation of large scale RNAi phenotypic datasets. AB - FLIGHT (www.flight.licr.org) is a new database designed to help researchers browse and cross-correlate data from large-scale RNAi studies. To date, the majority of these functional genomic screens have been carried out using Drosophila cell lines. These RNAi screens follow 100 years of classical Drosophila genetics, but have already revealed their potential by ascribing an impressive number of functions to known and novel genes. This has in turn given rise to a pressing need for tools to simplify the analysis of the large amount of phenotypic information generated. FLIGHT aims to do this by providing users with a gene-centric view of screen results and by making it possible to cluster phenotypic data to identify genes with related functions. Additionally, FLIGHT provides microarray expression data for many of the Drosophila cell lines commonly used in RNAi screens. This, together with information about cell lines, protocols and dsRNA primer sequences, is intended to help researchers design their own cell-based screens. Finally, although the current focus of FLIGHT is Drosophila, the database has been designed to facilitate the comparison of functional data across species and to help researchers working with other systems navigate their way through the fly genome. PMID- 16381917 TI - FlyBase: anatomical data, images and queries. AB - FlyBase (http://flybase.org/) is a database of genetic and genomic data on the model organism Drosophila melanogaster and the entire insect family Drosophilidae. The FlyBase Consortium curates, annotates, integrates and maintains a wide variety of data within this domain. Access to the data is provided through graphical and textual user interfaces tailored to particular types of data. FlyBase data types include maps at the cytological, genetic and sequence levels, genes and alleles including their products, functions, expression patterns, mutant phenotypes and genetic interactions as well as aberrant chromosomes, annotated genomes, genetic stock collections, transposons, transgene constructs and insertions, anatomy and images, bibliographic data, and community contact information. PMID- 16381918 TI - FlyRNAi: the Drosophila RNAi screening center database. AB - RNA interference (RNAi) has become a powerful tool for genetic screening in Drosophila. At the Drosophila RNAi Screening Center (DRSC), we are using a library of over 21,000 double-stranded RNAs targeting known and predicted genes in Drosophila. This library is available for the use of visiting scientists wishing to perform full-genome RNAi screens. The data generated from these screens are collected in the DRSC database (http://flyRNAi.org/cgi bin/RNAi_screens.pl) in a flexible format for the convenience of the scientist and for archiving data. The long-term goal of this database is to provide annotations for as many of the uncharacterized genes in Drosophila as possible. Data from published screens are available to the public through a highly configurable interface that allows detailed examination of the data and provides access to a number of other databases and bioinformatics tools. PMID- 16381919 TI - StellaBase: the Nematostella vectensis Genomics Database. AB - StellaBase, the Nematostella vectensis Genomics Database, is a web-based resource that will facilitate desktop and bench-top studies of the starlet sea anemone. Nematostella is an emerging model organism that has already proven useful for addressing fundamental questions in developmental evolution and evolutionary genomics. StellaBase allows users to query the assembled Nematostella genome, a confirmed gene library, and a predicted genome using both keyword and homology based search functions. Data provided by these searches will elucidate gene family evolution in early animals. Unique research tools, including a Nematostella genetic stock library, a primer library, a literature repository and a gene expression library will provide support to the burgeoning Nematostella research community. The development of StellaBase accompanies significant upgrades to CnidBase, the Cnidarian Evolutionary Genomics Database. With the completion of the first sequenced cnidarian genome, genome comparison tools have been added to CnidBase. In addition, StellaBase provides a framework for the integration of additional species-specific databases into CnidBase. StellaBase is available at http://www.stellabase.org. PMID- 16381920 TI - Tetrahymena Genome Database (TGD): a new genomic resource for Tetrahymena thermophila research. AB - We have developed a web-based resource (available at www.ciliate.org) for researchers studying the model ciliate organism Tetrahymena thermophila. Employing the underlying database structure and programming of the Saccharomyces Genome Database, the Tetrahymena Genome Database (TGD) integrates the wealth of knowledge generated by the Tetrahymena research community about genome structure, genes and gene products with the newly sequenced macronuclear genome determined by The Institute for Genomic Research (TIGR). TGD provides information curated from the literature about each published gene, including a standardized gene name, a link to the genomic locus in our graphical genome browser, gene product annotations utilizing the Gene Ontology, links to published literature about the gene and more. TGD also displays automatic annotations generated for the gene models predicted by TIGR. A variety of tools are available at TGD for searching the Tetrahymena genome, its literature and information about members of the research community. PMID- 16381921 TI - Pathguide: a pathway resource list. AB - Pathguide: the Pathway Resource List (http://pathguide.org) is a meta-database that provides an overview of more than 190 web-accessible biological pathway and network databases. These include databases on metabolic pathways, signaling pathways, transcription factor targets, gene regulatory networks, genetic interactions, protein-compound interactions, and protein-protein interactions. The listed databases are maintained by diverse groups in different locations and the information in them is derived either from the scientific literature or from systematic experiments. Pathguide is useful as a starting point for biological pathway analysis and for content aggregation in integrated biological information systems. PMID- 16381922 TI - LMPD: LIPID MAPS proteome database. AB - The LIPID MAPS Proteome Database (LMPD) is an object-relational database of lipid associated protein sequences and annotations. The initial release contains 2959 records, representing human and mouse proteins involved in lipid metabolism. UniProt IDs were obtained based on keyword search of KEGG and GO databases, and this LMPD protein list was then enhanced with annotations from UniProt, EntrezGene, ENZYME, GO, KEGG and other public resources. We also assigned associations with general lipid categories, based on GO and KEGG annotations. Users may search LMPD by database ID or keyword, and filter by species and/or lipid class associations; from the search results, one can then access a compilation of data relevant to each protein of interest, cross-linked to external databases. The LIPID MAPS Proteome Database (LMPD) is publicly available from the LIPID MAPS Consortium website (http://www.lipidmaps.org/). The direct URL is http://www.lipidmaps.org/data/proteome/index.cgi. PMID- 16381923 TI - MetaCyc: a multiorganism database of metabolic pathways and enzymes. AB - MetaCyc is a database of metabolic pathways and enzymes located at http://MetaCyc.org/. Its goal is to serve as a metabolic encyclopedia, containing a collection of non-redundant pathways central to small molecule metabolism, which have been reported in the experimental literature. Most of the pathways in MetaCyc occur in microorganisms and plants, although animal pathways are also represented. MetaCyc contains metabolic pathways, enzymatic reactions, enzymes, chemical compounds, genes and review-level comments. Enzyme information includes substrate specificity, kinetic properties, activators, inhibitors, cofactor requirements and links to sequence and structure databases. Data are curated from the primary literature by curators with expertise in biochemistry and molecular biology. MetaCyc serves as a readily accessible comprehensive resource on microbial and plant pathways for genome analysis, basic research, education, metabolic engineering and systems biology. Querying, visualization and curation of the database is supported by SRI's Pathway Tools software. The PathoLogic component of Pathway Tools is used in conjunction with MetaCyc to predict the metabolic network of an organism from its annotated genome. SRI and the European Bioinformatics Institute employed this tool to create pathway/genome databases (PGDBs) for 165 organisms, available at the BioCyc.org website. These PGDBs also include predicted operons and pathway hole fillers. PMID- 16381924 TI - The University of Minnesota Biocatalysis/Biodegradation Database: the first decade. AB - As the University of Minnesota Biocatalysis/Biodegradation Database (UM-BBD, http://umbbd.ahc.umn.edu/) starts its second decade, it includes information on over 900 compounds, over 600 enzymes, nearly 1000 reactions and about 350 microorganism entries. Its Biochemical Periodic Tables have grown to include biological information for almost all stable, non-noble-gas elements (http://umbbd.ahc.umn.edu/periodic/). Its Pathway Prediction System (PPS) (http://umbbd.ahc.umn.edu/predict/) is now an internationally recognized, open system for predicting microbial catabolism of organic compounds. Graphical display of PPS rules, a stand-alone version of the PPS and guidance for PPS users are being developed. The next decade should see the PPS, and the UM-BBD on which it is based, find increasing use by national and international government agencies, commercial organizations and educational institutions. PMID- 16381925 TI - AffinDB: a freely accessible database of affinities for protein-ligand complexes from the PDB. AB - AffinDB is a database of affinity data for structurally resolved protein-ligand complexes from the Protein Data Bank (PDB). It is freely accessible at http://www.agklebe.de/affinity. Affinity data are collected from the scientific literature, both from primary sources describing the original experimental work of affinity determination and from secondary references which report affinity values determined by others. AffinDB currently contains over 730 affinity entries covering more than 450 different protein-ligand complexes. Besides the affinity value, PDB summary information and additional data are provided, including the experimental conditions of the affinity measurement (if available in the corresponding reference); 2D drawing, SMILES code and molecular weight of the ligand; links to other databases, and bibliographic information. AffinDB can be queried by PDB code or by any combination of affinity range, temperature and pH value of the measurement, ligand molecular weight, and publication data (author, journal and year). Search results can be saved as tabular reports in text files. The database is supposed to be a valuable resource for researchers interested in biomolecular recognition and the development of tools for correlating structural data with affinities, as needed, for example, in structure-based drug design. PMID- 16381926 TI - pSTIING: a 'systems' approach towards integrating signalling pathways, interaction and transcriptional regulatory networks in inflammation and cancer. AB - pSTIING (http://pstiing.licr.org) is a new publicly accessible web-based application and knowledgebase featuring 65 228 distinct molecular associations (comprising protein-protein, protein-lipid, protein-small molecule interactions and transcriptional regulatory associations), ligand-receptor-cell type information and signal transduction modules. It has a particular major focus on regulatory networks relevant to chronic inflammation, cell migration and cancer. The web application and interface provide graphical representations of networks allowing users to combine and extend transcriptional regulatory and signalling modules, infer molecular interactions across species and explore networks via protein domains/motifs, gene ontology annotations and human diseases. pSTIING also supports the direct cross-correlation of experimental results with interaction information in the knowledgebase via the CLADIST tool associated with pSTIING, which currently analyses and clusters gene expression, proteomic and phenotypic datasets. This allows the contextual projection of co-expression patterns onto prior network information, facilitating the identification of functional modules in physiologically relevant systems. PMID- 16381927 TI - BioGRID: a general repository for interaction datasets. AB - Access to unified datasets of protein and genetic interactions is critical for interrogation of gene/protein function and analysis of global network properties. BioGRID is a freely accessible database of physical and genetic interactions available at http://www.thebiogrid.org. BioGRID release version 2.0 includes >116 000 interactions from Saccharomyces cerevisiae, Caenorhabditis elegans, Drosophila melanogaster and Homo sapiens. Over 30 000 interactions have recently been added from 5778 sources through exhaustive curation of the Saccharomyces cerevisiae primary literature. An internally hyper-linked web interface allows for rapid search and retrieval of interaction data. Full or user-defined datasets are freely downloadable as tab-delimited text files and PSI-MI XML. Pre-computed graphical layouts of interactions are available in a variety of file formats. User-customized graphs with embedded protein, gene and interaction attributes can be constructed with a visualization system called Osprey that is dynamically linked to the BioGRID. PMID- 16381928 TI - EndoNet: an information resource about endocrine networks. AB - EndoNet is a new database that provides information about the components of endocrine networks and their relations. It focuses on the endocrine cell-to-cell communication and enables the analysis of intercellular regulatory pathways in humans. In the EndoNet data model, two classes of components span a bipartite directed graph. One class represents the hormones (in the broadest sense) secreted by defined donor cells. The other class consists of the acceptor or target cells expressing the corresponding hormone receptors. The identity and anatomical environment of cell types, tissues and organs is defined through references to the CYTOMER ontology. With the EndoNet user interface, it is possible to query the database for hormones, receptors or tissues and to combine several items from different search rounds in one complex result set, from which a network can be reconstructed and visualized. For each entity, a detailed characteristics page is available. Some well-established endocrine pathways are offered as showcases in the form of predefined result sets. These sets can be used as a starting point for a more complex query or for obtaining a quick overview. The EndoNet database is accessible at http://endonet.bioinf.med.uni goettingen.de/. PMID- 16381929 TI - TRANSPATH: an information resource for storing and visualizing signaling pathways and their pathological aberrations. AB - TRANSPATH is a database about signal transduction events. It provides information about signaling molecules, their reactions and the pathways these reactions constitute. The representation of signaling molecules is organized in a number of orthogonal hierarchies reflecting the classification of the molecules, their species-specific or generic features, and their post-translational modifications. Reactions are similarly hierarchically organized in a three-layer architecture, differentiating between reactions that are evidenced by individual publications, generalizations of these reactions to construct species-independent 'reference pathways' and the 'semantic projections' of these pathways. A number of search and browse options allow easy access to the database contents, which can be visualized with the tool PathwayBuildertrade mark. The module PathoSign adds data about pathologically relevant mutations in signaling components, including their genotypes and phenotypes. TRANSPATH and PathoSign can be used as encyclopaedia, in the educational process, for vizualization and modeling of signal transduction networks and for the analysis of gene expression data. TRANSPATH Public 6.0 is freely accessible for users from non-profit organizations under http://www.gene regulation.com/pub/databases.html. PMID- 16381930 TI - DBTGR: a database of tunicate promoters and their regulatory elements. AB - The high similarity of tunicates and vertebrates during their development coupled with the transparency of tunicate larvae, their well-studied cell lineages and the availability of simple and efficient transgenesis methods makes of this subphylum an ideal system for the investigation of vertebrate physiological and developmental processes. Recently, the sequencing of two different Ciona genomes has lead to the identification of numerous genes. In order to better understand the regulation of these genes, a database was created containing information on regulation of tunicate genes collected from literature. It includes for instance information regarding the minimal promoter length, the transcription factors involved and their binding sites, as well as the localization of the gene expression. Additionally, binding sites for characterized transcription factors were predicted based on published in vitro recognition sites. Comparison of the promoters of homologous genes in different species is also provided to allow identification of conserved cis elements. At the time of writing, information about 184 promoters, containing 73 identified binding sites and >2000 newly predicted binding sites is available. This database is accessible at http://dbtgr.hgc.jp. PMID- 16381931 TI - Ensembl 2006. AB - The Ensembl (http://www.ensembl.org/) project provides a comprehensive and integrated source of annotation of large genome sequences. Over the last year the number of genomes available from the Ensembl site has increased from 4 to 19, with the addition of the mammalian genomes of Rhesus macaque and Opossum, the chordate genome of Ciona intestinalis and the import and integration of the yeast genome. The year has also seen extensive improvements to both data analysis and presentation, with the introduction of a redesigned website, the addition of RNA gene and regulatory annotation and substantial improvements to the integration of human genome variation data. PMID- 16381932 TI - HOLLYWOOD: a comparative relational database of alternative splicing. AB - RNA splicing is an essential step in gene expression, and is often variable, giving rise to multiple alternatively spliced mRNA and protein isoforms from a single gene locus. The design of effective databases to support experimental and computational investigations of alternative splicing (AS) is a significant challenge. In an effort to integrate accurate exon and splice site annotation with current knowledge about splicing regulatory elements and predicted AS events, and to link information about the splicing of orthologous genes in different species, we have developed the Hollywood system. This database was built upon genomic annotation of splicing patterns of known genes derived from spliced alignment of complementary DNAs (cDNAs) and expressed sequence tags, and links features such as splice site sequence and strength, exonic splicing enhancers and silencers, conserved and non-conserved patterns of splicing, and cDNA library information for inferred alternative exons. Hollywood was implemented as a relational database and currently contains comprehensive information for human and mouse. It is accompanied by a web query tool that allows searches for sets of exons with specific splicing characteristics or splicing regulatory element composition, or gives a graphical or sequence-level summary of splicing patterns for a specific gene. A streamlined graphical representation of gene splicing patterns is provided, and these patterns can alternatively be layered onto existing information in the UCSC Genome Browser. The database is accessible at http://hollywood.mit.edu. PMID- 16381933 TI - The Mouse Genome Database (MGD): updates and enhancements. AB - The Mouse Genome Database (MGD) integrates genetic and genomic data for the mouse in order to facilitate the use of the mouse as a model system for understanding human biology and disease processes. A core component of the MGD effort is the acquisition and integration of genomic, genetic, functional and phenotypic information about mouse genes and gene products. MGD works within the broader bioinformatics community to define referential and semantic standards to facilitate data exchange between resources including the incorporation of information from the biomedical literature. MGD is also a platform for computational assessment of integrated biological data with the goal of identifying candidate genes associated with complex phenotypes. MGD is web accessible at http://www.informatics.jax.org. Recent improvements in MGD described here include the incorporation of an interactive genome browser, the enhancement of phenotype resources and the further development of functional annotation resources. PMID- 16381934 TI - The Mouse Functional Genome Database (MfunGD): functional annotation of proteins in the light of their cellular context. AB - MfunGD (http://mips.gsf.de/genre/proj/mfungd/) provides a resource for annotated mouse proteins and their occurrence in protein networks. Manual annotation concentrates on proteins which are found to interact physically with other proteins. Accordingly, manually curated information from a protein-protein interaction database (MPPI) and a database of mammalian protein complexes is interconnected with MfunGD. Protein function annotation is performed using the Functional Catalogue (FunCat) annotation scheme which is widely used for the analysis of protein networks. The dataset is also supplemented with information about the literature that was used in the annotation process as well as links to the SIMAP Fasta database, the Pedant protein analysis system and cross-references to external resources. Proteins that so far were not manually inspected are annotated automatically by a graphical probabilistic model and/or superparamagnetic clustering. The database is continuously expanding to include the rapidly growing amount of functional information about gene products from mouse. MfunGD is implemented in GenRE, a J2EE-based component-oriented multi-tier architecture following the separation of concern principle. PMID- 16381935 TI - TreeFam: a curated database of phylogenetic trees of animal gene families. AB - TreeFam is a database of phylogenetic trees of gene families found in animals. It aims to develop a curated resource that presents the accurate evolutionary history of all animal gene families, as well as reliable ortholog and paralog assignments. Curated families are being added progressively, based on seed alignments and trees in a similar fashion to Pfam. Release 1.1 of TreeFam contains curated trees for 690 families and automatically generated trees for another 11 646 families. These represent over 128 000 genes from nine fully sequenced animal genomes and over 45 000 other animal proteins from UniProt; approximately 40-85% of proteins encoded in the fully sequenced animal genomes are included in TreeFam. TreeFam is freely available at http://www.treefam.org and http://treefam.genomics.org.cn. PMID- 16381936 TI - The Zebrafish Information Network: the zebrafish model organism database. AB - The Zebrafish Information Network (ZFIN; http://zfin.org) is a web based community resource that implements the curation of zebrafish genetic, genomic and developmental data. ZFIN provides an integrated representation of mutants, genes, genetic markers, mapping panels, publications and community resources such as meeting announcements and contact information. Recent enhancements to ZFIN include (i) comprehensive curation of gene expression data from the literature and from directly submitted data, (ii) increased support and annotation of the genome sequence, (iii) expanded use of ontologies to support curation and query forms, (iv) curation of morpholino data from the literature, and (v) increased versatility of gene pages, with new data types, links and analysis tools. PMID- 16381937 TI - QTL MatchMaker: a multi-species quantitative trait loci (QTL) database and query system for annotation of genes and QTL. AB - Identifying genes that underlie quantitative trait loci (QTL) is a challenging task. Here, we present a new QTL software system, named QTL MatchMaker. The system is designed to integrate and mine QTL information across human, mouse and rat genomes and to annotate functional genomic data. It combines and organizes information from relevant public databases and publications and integrates QTL, physical, genetic and cytogenetic maps across human, mouse and rat. To make this application available to the research community we have developed a website for high-throughput mapping of expressed sequences to QTL and for selection of candidate genes in the physiological genomics context of complex traits. QTL MatchMaker is accessible at http://pmrc.med.mssm.edu:9090/QTL/jsp/qtlhome.jsp. PMID- 16381938 TI - The UCSC Genome Browser Database: update 2006. AB - The University of California Santa Cruz Genome Browser Database (GBD) contains sequence and annotation data for the genomes of about a dozen vertebrate species and several major model organisms. Genome annotations typically include assembly data, sequence composition, genes and gene predictions, mRNA and expressed sequence tag evidence, comparative genomics, regulation, expression and variation data. The database is optimized to support fast interactive performance with web tools that provide powerful visualization and querying capabilities for mining the data. The Genome Browser displays a wide variety of annotations at all scales from single nucleotide level up to a full chromosome. The Table Browser provides direct access to the database tables and sequence data, enabling complex queries on genome-wide datasets. The Proteome Browser graphically displays protein properties. The Gene Sorter allows filtering and comparison of genes by several metrics including expression data and several gene properties. BLAT and In Silico PCR search for sequences in entire genomes in seconds. These tools are highly integrated and provide many hyperlinks to other databases and websites. The GBD, browsing tools, downloadable data files and links to documentation and other information can be found at http://genome.ucsc.edu/. PMID- 16381939 TI - OMIA (Online Mendelian Inheritance in Animals): an enhanced platform and integration into the Entrez search interface at NCBI. AB - Online Mendelian Inheritance in Animals (OMIA) is a comprehensive, annotated catalogue of inherited disorders and other familial traits in animals other than humans and mice. Structured as a comparative biology resource, OMIA is a comprehensive resource of phenotypic information on heritable animal traits and genes in a strongly comparative context, relating traits to genes where possible. OMIA is modelled on and is complementary to Online Mendelian Inheritance in Man (OMIM). OMIA has been moved to a MySQL database at the Australian National Genomic Information Service (ANGIS) and can be accessed at http://omia.angis.org.au/. It has also been integrated into the Entrez search interface at the National Center for Biotechnology Information (NCBI; http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=omia). Curation of OMIA data by researchers working on particular species and disorders has also been enabled. PMID- 16381940 TI - DDBJ in preparation for overview of research activities behind data submissions. AB - In the past year, DDBJ (http://www.ddbj.nig.ac.jp) collected and released 1,956,826 entries or 1,741,313,111 bases. The released data include approximately 90,000 ESTs and cDNAs of Macaca fascicularis, and 280 million bases of mouse GSS. In addition to the data collection, we have indexed the submitted data to the International Nucleotide Sequence Database Collaboration (INSDC, http://www.insdc.org) to classify the entries into research projects behind data submissions. They are expected to be useful to the data submitters and users for enhancing the data submission, retrieval and systematic data analyses at INSDC. The results of indexing also allow one to grasp research projects in life sciences that promoted and produced the DNA sequences submitted to INSDC. PMID- 16381941 TI - CTGA: the database for genetic disorders in Arab populations. AB - The Arabs comprise a genetically heterogeneous group that resulted from the admixture of different populations throughout history. They share many common characteristics responsible for a considerable proportion of perinatal and neonatal mortalities. To this end, the Centre for Arab Genomic Studies (CAGS) launched a pilot project to construct the 'Catalogue of Transmission Genetics in Arabs' (CTGA) database for genetic disorders in Arabs. Information in CTGA is drawn from published research and mined hospital records. The database offers web based basic and advanced search approaches. In either case, the final search result is a detailed HTML record that includes text-, URL- and graphic-based fields. At present, CTGA hosts entries for 692 phenotypes and 235 related genes described in Arab individuals. Of these, 213 phenotypic descriptions and 22 related genes were observed in the Arab population of the United Arab Emirates (UAE). These results emphasize the role of CTGA as an essential tool to promote scientific research on genetic disorders in the region. The priority of CTGA is to provide timely information on the occurrence of genetic disorders in Arab individuals. It is anticipated that data from Arab countries other than the UAE will be exhaustively searched and incorporated in CTGA (http://www.cags.org.ae). PMID- 16381942 TI - HPtaa database-potential target genes for clinical diagnosis and immunotherapy of human carcinoma. AB - Tumor-associated antigens (TAAs) have been the most actively employed targets in the clinical diagnosis and treatment of human carcinoma, such as PSA in the diagnosis of prostate cancer and NY-ESO-1 in the immunotherapy of melanoma and other cancers. However, identification of TAAs has often been hampered by the complicated and laborsome laboratory procedures. In order to accelerate the process of tumor antigen discovery, and thereby improve diagnosis and treatment of human carcinoma, we have made an effort to establish a publicly available Human Potential Tumor Associated Antigen database (HPtaa) with potential TAAs identified by in silico computing (http://www.hptaa.org). Tumor specificity was chosen as the core of tumor antigen evaluation, together with other relevant clues. Various platforms of gene expression, including microarray, expressed sequence tag and SAGE data, were processed and integrated by several penalty algorithms. A total of 3518 potential TAAs have been included in the database, which is freely available to academic users. As far as we know, this database is the first one addressing human potential TAAs, and the first one integrating various kinds of expression platforms for one purpose. PMID- 16381943 TI - ITTACA: a new database for integrated tumor transcriptome array and clinical data analysis. AB - Transcriptome microarrays have become one of the tools of choice for investigating the genes involved in tumorigenesis and tumor progression, as well as finding new biomarkers and gene expression signatures for the diagnosis and prognosis of cancer. Here, we describe a new database for Integrated Tumor Transcriptome Array and Clinical data Analysis (ITTACA). ITTACA centralizes public datasets containing both gene expression and clinical data. ITTACA currently focuses on the types of cancer that are of particular interest to research teams at Institut Curie: breast carcinoma, bladder carcinoma and uveal melanoma. A web interface allows users to carry out different class comparison analyses, including the comparison of expression distribution profiles, tests for differential expression and patient survival analyses. ITTACA is complementary to other databases, such as GEO and SMD, because it offers a better integration of clinical data and different functionalities. It also offers more options for class comparison analyses when compared with similar projects such as Oncomine. For example, users can define their own patient groups according to clinical data or gene expression levels. This added flexibility and the user-friendly web interface makes ITTACA especially useful for comparing personal results with the results in the existing literature. ITTACA is accessible online at http://bioinfo.curie.fr/ittaca. PMID- 16381944 TI - SNP500Cancer: a public resource for sequence validation, assay development, and frequency analysis for genetic variation in candidate genes. AB - The SNP500Cancer database provides sequence and genotype assay information for candidate SNPs useful in mapping complex diseases, such as cancer. The database is an integral component of the NCI Cancer Genome Anatomy Project (http://cgap.nci.nih.gov). SNP500Cancer reports sequence analysis of anonymized control DNA samples (n = 102 Coriell samples representing four self-described ethnic groups: African/African-American, Caucasian, Hispanic and Pacific Rim). The website is searchable by gene, chromosome, gene ontology pathway, dbSNP ID and SNP500Cancer SNP ID. As of October 2005, the database contains >13 400 SNPs, 9124 of which have been sequenced in the SNP500Cancer population. For each analysed SNP, gene location and >200 bp of surrounding annotated sequence (including nearby SNPs) are provided, with frequency information in total and per subpopulation as well as calculation of Hardy-Weinberg equilibrium for each subpopulation. The website provides the conditions for validated sequencing and genotyping assays, as well as genotype results for the 102 samples, in both viewable and downloadable formats. A subset of sequence validated SNPs with minor allele frequency >5% are entered into a high-throughput pipeline for genotyping analysis to determine concordance for the same 102 samples. In addition, the results of genotype analysis for select validated SNP assays (defined as 100% concordance between sequence analysis and genotype results) are posted for an additional 280 samples drawn from the Human Diversity Panel (HDP). SNP500Cancer provides an invaluable resource for investigators to select SNPs for analysis, design genotyping assays using validated sequence data, choose selected assays already validated on one or more genotyping platforms, and select reference standards for genotyping assays. The SNP500Cancer database is freely accessible via the web page at http://snp500cancer.nci.nih.gov. PMID- 16381945 TI - dbPTM: an information repository of protein post-translational modification. AB - dbPTM is a database that compiles information on protein post-translational modifications (PTMs), such as the catalytic sites, solvent accessibility of amino acid residues, protein secondary and tertiary structures, protein domains and protein variations. The database includes all of the experimentally validated PTM sites from Swiss-Prot, PhosphoELM and O-GLYCBASE. Only a small fraction of Swiss Prot proteins are annotated with experimentally verified PTM. Although the Swiss Prot provides rich information about the PTM, other structural properties and functional information of proteins are also essential for elucidating protein mechanisms. The dbPTM systematically identifies three major types of protein PTM (phosphorylation, glycosylation and sulfation) sites against Swiss-Prot proteins by refining our previously developed prediction tool, KinasePhos (http://kinasephos.mbc.nctu.edu.tw/). Solvent accessibility and secondary structure of residues are also computationally predicted and are mapped to the PTM sites. The resource is now freely available at http://dbPTM.mbc.nctu.edu.tw/. PMID- 16381946 TI - BodyMap-Xs: anatomical breakdown of 17 million animal ESTs for cross-species comparison of gene expression. AB - BodyMap-Xs (http://bodymap.jp) is a database for cross-species gene expression comparison. It was created by the anatomical breakdown of 17 million animal expressed sequence tag (EST) records in DDBJ using a sorting program tailored for this purpose. In BodyMap-Xs, users are allowed to compare the expression patterns of orthologous and paralogous genes in a coherent manner. This will provide valuable insights for the evolutionary study of gene expression and identification of a responsive motif for a particular expression pattern. In addition, starting from a concise overview of the taxonomical and anatomical breakdown of all animal ESTs, users can navigate to obtain gene expression ranking of a particular tissue in a particular animal. This method may lead to the understanding of the similarities and differences between the homologous tissues across animal species. BodyMap-Xs will be automatically updated in synchronization with the major update in DDBJ, which occurs periodically. PMID- 16381947 TI - ABS: a database of Annotated regulatory Binding Sites from orthologous promoters. AB - Information about the genomic coordinates and the sequence of experimentally identified transcription factor binding sites is found scattered under a variety of diverse formats. The availability of standard collections of such high-quality data is important to design, evaluate and improve novel computational approaches to identify binding motifs on promoter sequences from related genes. ABS (http://genome.imim.es/datasets/abs2005/index.html) is a public database of known binding sites identified in promoters of orthologous vertebrate genes that have been manually curated from bibliography. We have annotated 650 experimental binding sites from 68 transcription factors and 100 orthologous target genes in human, mouse, rat or chicken genome sequences. Computational predictions and promoter alignment information are also provided for each entry. A simple and easy-to-use web interface facilitates data retrieval allowing different views of the information. In addition, the release 1.0 of ABS includes a customizable generator of artificial datasets based on the known sites contained in the collection and an evaluation tool to aid during the training and the assessment of motif-finding programs. PMID- 16381948 TI - CAGE Basic/Analysis Databases: the CAGE resource for comprehensive promoter analysis. AB - Cap-analysis gene expression (CAGE) Basic and Analysis Databases store an original resource produced by CAGE, which measures expression levels of transcription starting sites by sequencing large amounts of transcript 5' ends, termed CAGE tags. Millions of human and mouse high-quality CAGE tags derived from different conditions in >20 tissues consisting of >250 RNA samples are essential for identification of novel promoters and promoter characterization in the aspect of expression profile. CAGE Basic Database is a primary database of the CAGE resource, RNA samples, CAGE libraries, CAGE clone and tag sequences and so on. CAGE Analysis Database stores promoter related information, such as counts of related transcripts, CpG islands and conserved genome region. It also provides expression profiles at base pair and promoter levels. Both databases are based on the same framework, CAGE tag starting sites, tag clusters for defining promoters and transcriptional units (TUs). Their associations and TU attributes are available to find promoters of interest. These databases were provided for Functional Annotation Of Mouse 3 (FANTOM3), an international collaboration research project focusing on expanding the transcriptome and subsequent analyses. Now access is free for all users through the World Wide Web at http://fantom3.gsc.riken.jp/. PMID- 16381949 TI - EMAGE: a spatial database of gene expression patterns during mouse embryo development. AB - EMAGE (http://genex.hgu.mrc.ac.uk/Emage/database) is a freely available, curated database of gene expression patterns generated by in situ techniques in the developing mouse embryo. It is unique in that it contains standardized spatial representations of the sites of gene expression for each gene, denoted against a set of virtual reference embryo models. As such, the data can be interrogated in a novel and abstract manner by using space to define a query. Accompanying the spatial representations of gene expression patterns are text descriptions of the sites of expression, which also allows searching of the data by more conventional text-based methods. PMID- 16381950 TI - The International Gene Trap Consortium Website: a portal to all publicly available gene trap cell lines in mouse. AB - Gene trapping is a method of generating murine embryonic stem (ES) cell lines containing insertional mutations in known and novel genes. A number of international groups have used this approach to create sizeable public cell line repositories available to the scientific community for the generation of mutant mouse strains. The major gene trapping groups worldwide have recently joined together to centralize access to all publicly available gene trap lines by developing a user-oriented Website for the International Gene Trap Consortium (IGTC). This collaboration provides an impressive public informatics resource comprising approximately 45 000 well-characterized ES cell lines which currently represent approximately 40% of known mouse genes, all freely available for the creation of knockout mice on a non-collaborative basis. To standardize annotation and provide high confidence data for gene trap lines, a rigorous identification and annotation pipeline has been developed combining genomic localization and transcript alignment of gene trap sequence tags to identify trapped loci. This information is stored in a new bioinformatics database accessible through the IGTC Website interface. The IGTC Website (www.genetrap.org) allows users to browse and search the database for trapped genes, BLAST sequences against gene trap sequence tags, and view trapped genes within biological pathways. In addition, IGTC data have been integrated into major genome browsers and bioinformatics sites to provide users with outside portals for viewing this data. The development of the IGTC Website marks a major advance by providing the research community with the data and tools necessary to effectively use public gene trap resources for the large-scale characterization of mammalian gene function. PMID- 16381951 TI - PepSeeker: a database of proteome peptide identifications for investigating fragmentation patterns. AB - Proteome science relies on bioinformatics tools to characterize proteins via their proteolytic peptides which are identified via characteristic mass spectra generated after their ions undergo fragmentation in the gas phase within the mass spectrometer. The resulting secondary ion mass spectra are compared with protein sequence databases in order to identify the amino acid sequence. Although these search tools (e.g. SEQUEST, Mascot, X!Tandem, Phenyx) are frequently successful, much is still not understood about the amino acid sequence patterns which promote/protect particular fragmentation pathways, and hence lead to the presence/absence of particular ions from different ion series. In order to advance this area, we have developed a database, PepSeeker (http://nwsr.smith.man.ac.uk/pepseeker), which captures this peptide identification and ion information from proteome experiments. The database currently contains >185,000 peptides and associated database search information. Users may query this resource to retrieve peptide, protein and spectral information based on protein or peptide information, including the amino acid sequence itself represented by regular expressions coupled with ion series information. We believe this database will be useful to proteome researchers wishing to understand gas phase peptide ion chemistry in order to improve peptide identification strategies. Questions can be addressed to j.selley@manchester.ac.uk. PMID- 16381952 TI - The PeptideAtlas project. AB - The completion of the sequencing of the human genome and the concurrent, rapid development of high-throughput proteomic methods have resulted in an increasing need for automated approaches to archive proteomic data in a repository that enables the exchange of data among researchers and also accurate integration with genomic data. PeptideAtlas (http://www.peptideatlas.org/) addresses these needs by identifying peptides by tandem mass spectrometry (MS/MS), statistically validating those identifications and then mapping identified sequences to the genomes of eukaryotic organisms. A meaningful comparison of data across different experiments generated by different groups using different types of instruments is enabled by the implementation of a uniform analytic process. This uniform statistical validation ensures a consistent and high-quality set of peptide and protein identifications. The raw data from many diverse proteomic experiments are made available in the associated PeptideAtlas repository in several formats. Here we present a summary of our process and details about the Human, Drosophila and Yeast PeptideAtlas builds. PMID- 16381953 TI - PRIDE: a public repository of protein and peptide identifications for the proteomics community. AB - PRIDE, the 'PRoteomics IDEntifications database' (http://www.ebi.ac.uk/pride) is a database of protein and peptide identifications that have been described in the scientific literature. These identifications will typically be from specific species, tissues and sub-cellular locations, perhaps under specific disease conditions. Any post-translational modifications that have been identified on individual peptides can be described. These identifications may be annotated with supporting mass spectra. At the time of writing, PRIDE includes the full set of identifications as submitted by individual laboratories participating in the HUPO Plasma Proteome Project and a profile of the human platelet proteome submitted by the University of Ghent in Belgium. By late 2005 PRIDE is expected to contain the identifications and spectra generated by the HUPO Brain Proteome Project. Proteomics laboratories are encouraged to submit their identifications and spectra to PRIDE to support their manuscript submissions to proteomics journals. Data can be submitted in PRIDE XML format if identifications are included or mzData format if the submitter is depositing mass spectra without identifications. PRIDE is a web application, so submission, searching and data retrieval can all be performed using an internet browser. PRIDE can be searched by experiment accession number, protein accession number, literature reference and sample parameters including species, tissue, sub-cellular location and disease state. Data can be retrieved as machine-readable PRIDE or mzData XML (the latter for mass spectra without identifications), or as human-readable HTML. PMID- 16381954 TI - AOBase: a database for antisense oligonucleotides selection and design. AB - Antisense oligonucleotides (ODNs) technology is one of the important approaches for the sequence-specific knockdown of gene expression. ODNs have been used as research tools in the post-genome era, as well as new types of therapeutic agents. Since finding effective target sites within RNA is a hard work for antisense ODNs design, various experimental methods and computational approaches have been proposed. For better sharing of the experimented and published ODNs, valid and invalid ODNs reported in literatures are screened, collected and stored in AOBase. Till now, approximately 700 ODNs against 46 target mRNAs are contained in AOBase. Entries can be explored via TargetSearch and AOSearch web retrieval interfaces. AOBase can not only be useful in ODNs selection for gene function exploration, but also contribute to mining rules and developing algorithms for rational ODNs design. AOBase is freely accessible via http://www.bioit.org.cn/ao/aobase. PMID- 16381955 TI - DrugBank: a comprehensive resource for in silico drug discovery and exploration. AB - DrugBank is a unique bioinformatics/cheminformatics resource that combines detailed drug (i.e. chemical) data with comprehensive drug target (i.e. protein) information. The database contains >4100 drug entries including >800 FDA approved small molecule and biotech drugs as well as >3200 experimental drugs. Additionally, >14,000 protein or drug target sequences are linked to these drug entries. Each DrugCard entry contains >80 data fields with half of the information being devoted to drug/chemical data and the other half devoted to drug target or protein data. Many data fields are hyperlinked to other databases (KEGG, PubChem, ChEBI, PDB, Swiss-Prot and GenBank) and a variety of structure viewing applets. The database is fully searchable supporting extensive text, sequence, chemical structure and relational query searches. Potential applications of DrugBank include in silico drug target discovery, drug design, drug docking or screening, drug metabolism prediction, drug interaction prediction and general pharmaceutical education. DrugBank is available at http://redpoll.pharmacy.ualberta.ca/drugbank/. PMID- 16381956 TI - GLIDA: GPCR-ligand database for chemical genomic drug discovery. AB - G-protein coupled receptors (GPCRs) represent one of the most important families of drug targets in pharmaceutical development. GPCR-LIgand DAtabase (GLIDA) is a novel public GPCR-related chemical genomic database that is primarily focused on the correlation of information between GPCRs and their ligands. It provides correlation data between GPCRs and their ligands, along with chemical information on the ligands, as well as access information to the various web databases regarding GPCRs. These data are connected with each other in a relational database, allowing users in the field of GPCR-related drug discovery to easily retrieve such information from either biological or chemical starting points. GLIDA includes structure similarity search functions for the GPCRs and for their ligands. Thus, GLIDA can provide correlation maps linking the searched homologous GPCRs (or ligands) with their ligands (or GPCRs). By analyzing the correlation patterns between GPCRs and ligands, we can gain more detailed knowledge about their interactions and improve drug design efforts by focusing on inferred candidates for GPCR-specific drugs. GLIDA is publicly available at http://gdds.pharm.kyoto-u.ac.jp:8081/glida. We hope that it will prove very useful for chemical genomic research and GPCR-related drug discovery. PMID- 16381957 TI - SuperNatural: a searchable database of available natural compounds. AB - Although tremendous effort has been put into synthetic libraries, most drugs on the market are still natural compounds or derivatives thereof. There are encyclopaedias of natural compounds, but the availability of these compounds is often unclear and catalogues from numerous suppliers have to be checked. To overcome these problems we have compiled a database of approximately 50,000 natural compounds from different suppliers. To enable efficient identification of the desired compounds, we have implemented substructure searches with typical templates. Starting points for in silico screenings are about 2500 well-known and classified natural compounds from a compendium that we have added. Possible medical applications can be ascertained via automatic searches for similar drugs in a free conformational drug database containing WHO indications. Furthermore, we have computed about three million conformers, which are deployed to account for the flexibilities of the compounds when the 3D superposition algorithm that we have developed is used. The SuperNatural Database is publicly available at http://bioinformatics.charite.de/supernatural. Viewing requires the free Chime plugin from MDL (Chime) or Java2 Runtime Environment (MView), which is also necessary for using Marvin application for chemical drawing. PMID- 16381958 TI - cisRED: a database system for genome-scale computational discovery of regulatory elements. AB - We describe cisRED, a database for conserved regulatory elements that are identified and ranked by a genome-scale computational system (www.cisred.org). The database and high-throughput predictive pipeline are designed to address diverse target genomes in the context of rapidly evolving data resources and tools. Motifs are predicted in promoter regions using multiple discovery methods applied to sequence sets that include corresponding sequence regions from vertebrates. We estimate motif significance by applying discovery and post processing methods to randomized sequence sets that are adaptively derived from target sequence sets, retain motifs with p-values below a threshold and identify groups of similar motifs and co-occurring motif patterns. The database offers information on atomic motifs, motif groups and patterns. It is web-accessible, and can be queried directly, downloaded or installed locally. PMID- 16381959 TI - RTPrimerDB: the real-time PCR primer and probe database, major update 2006. AB - The RTPrimerDB (http://medgen.ugent.be/rtprimerdb) project provides a freely accessible data retrieval system and an in silico assay evaluation pipeline for real-time quantitative PCR assays. Over the last year the number of user submitted assays has grown to 3500. Data conveyance from Entrez Gene by establishing an assay-to-gene relationship enables the addition of new primer assays for one of the 1.5 million different genes from 2300 species stored in the system. Easy access to the primer and probe data is possible by using multiple search criteria. Assay reports contain gene information, assay details (such as oligonucleotide sequences, detection chemistry and reaction conditions), publication information, users' experimental evaluation feedback and submitter's contact details. Gene expression assays are extended with a scalable assay viewer that provides detailed information on the alignment of primer and probe sequences on the known transcript variants of a gene, along with Single Nucleotide Polymorphisms (SNP) positions and peptide domain information. Furthermore, an mfold module is implemented to predict the secondary structure of the amplicon sequence, as this has been reported to impact the efficiency of the PCR. RTPrimerDB is also extended with an in silico analysis pipeline to streamline the evaluation of custom designed primer and probe sequences prior to ordering and experimental evaluation. In a secured environment, the pipeline performs automated BLAST specificity searches, mfold secondary structure prediction, SNP or plain sequence error identification, and graphical visualization of the aligned primer and probe sequences on the target gene. PMID- 16381960 TI - BioModels Database: a free, centralized database of curated, published, quantitative kinetic models of biochemical and cellular systems. AB - BioModels Database (http://www.ebi.ac.uk/biomodels/), part of the international initiative BioModels.net, provides access to published, peer-reviewed, quantitative models of biochemical and cellular systems. Each model is carefully curated to verify that it corresponds to the reference publication and gives the proper numerical results. Curators also annotate the components of the models with terms from controlled vocabularies and links to other relevant data resources. This allows the users to search accurately for the models they need. The models can currently be retrieved in the SBML format, and import/export facilities are being developed to extend the spectrum of formats supported by the resource. PMID- 16381961 TI - ChloroplastDB: the Chloroplast Genome Database. AB - The Chloroplast Genome Database (ChloroplastDB) is an interactive, web-based database for fully sequenced plastid genomes, containing genomic, protein, DNA and RNA sequences, gene locations, RNA-editing sites, putative protein families and alignments (http://chloroplast.cbio.psu.edu/). With recent technical advances, the rate of generating new organelle genomes has increased dramatically. However, the established ontology for chloroplast genes and gene features has not been uniformly applied to all chloroplast genomes available in the sequence databases. For example, annotations for some published genome sequences have not evolved with gene naming conventions. ChloroplastDB provides unified annotations, gene name search, BLAST and download functions for chloroplast encoded genes and genomic sequences. A user can retrieve all orthologous sequences with one search regardless of gene names in GenBank. This feature alone greatly facilitates comparative research on sequence evolution including changes in gene content, codon usage, gene structure and post transcriptional modifications such as RNA editing. Orthologous protein sets are classified by TribeMCL and each set is assigned a standard gene name. Over the next few years, as the number of sequenced chloroplast genomes increases rapidly, the tools available in ChloroplastDB will allow researchers to easily identify and compile target data for comparative analysis of chloroplast genes and genomes. PMID- 16381962 TI - GOBASE--a database of organelle and bacterial genome information. AB - The organelle genome database GOBASE is now in its twelfth release, and includes 350,000 mitochondrial sequences and 118,000 chloroplast sequences, roughly a 3 fold expansion since previously documented. GOBASE also includes a fully reannotated genome sequence of Rickettsia prowazekii, one of the closest bacterial relatives of mitochondria, and will shortly expand to contain more data from bacteria from which organelles originated. All these sequences are now accessible through a single unified interface. Enhancements to the functionality of GOBASE include addition of pages for RNA structures and a page compiling data about the taxonomic distribution of organelle-encoded genes; incorporation of Gene Ontology terms; addition of features deduced from incomplete annotations to sequences in GenBank; marking of type examples in cases where single genes in single species are oversampled within GenBank; and addition of graphics illustrating gene structure and the position of neighbouring genes on a sequence. The database has been reimplemented in PostgreSQL to facilitate development and maintenance, and structural modifications have been made to speed up queries, particularly those related to taxonomy. The GOBASE database can be queried at http://gobase.bcm.umontreal.ca/ and inquiries should be directed to gobase@bch.umontreal.ca. PMID- 16381963 TI - HvrBase++: a phylogenetic database for primate species. AB - HvrBase++ is the improved and extended version of HvrBase. Extensions are made by adding more population-based sequence samples from all primates including humans. The current collection comprises 13,873 hypervariable region I (HVRI) sequences and 4940 hypervariable region II (HVRII) sequences. In addition, we included 1376 complete mitochondrial genomes, 205 sequences from X-chromosomal loci and 202 sequences from autosomal chromosomes 1, 8, 11 and 16. In order to reduce the introduction of erroneous data into HvrBase++, we have developed a procedure that monitors GenBank for new versions of the current data in HvrBase++ and automatically updates the collection if necessary. For the stored sequences, supplementary information such as geographic origin, population affiliation and language of the sequence donor can be retrieved. HvrBase++ is Oracle based and easily accessible by a web interface (http://www.hvrbase.org). As a new key feature, HvrBase++ provides an interactive graphical tool to easily access data from dynamically created geographical maps. PMID- 16381964 TI - MitoP2: the mitochondrial proteome database--now including mouse data. AB - The MitoP2 database (http://www.mitop.de) integrates information on mitochondrial proteins, their molecular functions and associated diseases. The central database features are manually annotated reference proteins localized or functionally associated with mitochondria supplied for yeast, human and mouse. MitoP2 enables (i) the identification of putative orthologous proteins between these species to study evolutionarily conserved functions and pathways; (ii) the integration of data from systematic genome-wide studies such as proteomics and deletion phenotype screening; (iii) the prediction of novel mitochondrial proteins using data integration and the assignment of evidence scores; and (iv) systematic searches that aim to find the genes that underlie common and rare mitochondrial diseases. The data and analysis files are referenced to data sources in PubMed and other online databases and can be easily downloaded. MitoP2 users can explore the relationship between mitochondrial dysfunctions and disease and utilize this information to conduct systems biology approaches on mitochondria. PMID- 16381965 TI - DRASTIC--INSIGHTS: querying information in a plant gene expression database. AB - DRASTIC--Database Resource for the Analysis of Signal Transduction In Cells (http://www.drastic.org.uk/) has been created as a first step towards a data based approach for constructing signal transduction pathways. DRASTIC is a relational database of plant expressed sequence tags and genes up- or down regulated in response to various pathogens, chemical exposure or other treatments such as drought, salt and low temperature. More than 17700 records have been obtained from 306 treatments affecting 73 plant species from 512 peer-reviewed publications with most emphasis being placed on data from Arabidopsis thaliana. DRASTIC has been developed by the Scottish Crop Research Institute and the University of Abertay Dundee and allows rapid identification of plant genes that are up- or down-regulated by multiple treatments and those that are regulated by a very limited (or perhaps a single) treatment. The INSIGHTS (INference of cell SIGnaling HypoTheseS) suite of web-based tools allows intelligent data mining and extraction of information from the DRASTIC database. Potential response pathways can be visualized and comparisons made between gene expression patterns in response to various treatments. The knowledge gained informs plant signalling pathways and systems biology investigations. PMID- 16381966 TI - Gramene: a bird's eye view of cereal genomes. AB - Rice, maize, sorghum, wheat, barley and the other major crop grasses from the family Poaceae (Gramineae) are mankind's most important source of calories and contribute tens of billions of dollars annually to the world economy (FAO 1999, http://www.fao.org; USDA 1997, http://www.usda.gov). Continued improvement of Poaceae crops is necessary in order to continue to feed an ever-growing world population. However, of the major crop grasses, only rice (Oryza sativa), with a compact genome of approximately 400 Mbp, has been sequenced and annotated. The Gramene database (http://www.gramene.org) takes advantage of the known genetic colinearity (synteny) between rice and the major crop plant genomes to provide maize, sorghum, millet, wheat, oat and barley researchers with the benefits of an annotated genome years before their own species are sequenced. Gramene is a one stop portal for finding curated literature, genetic and genomic datasets related to maps, markers, genes, genomes and quantitative trait loci. The addition of several new tools to Gramene has greatly facilitated the potential for comparative analysis among the grasses and contributes to our understanding of the anatomy, development, environmental responses and the factors influencing agronomic performance of cereal crops. Since the last publication on Gramene database by D. H. Ware, P. Jaiswal, J. Ni, I. V. Yap, X. Pan, K. Y. Clark, L. Teytelman, S. C. Schmidt, W. Zhao, K. Chang et al. [(2002), Plant Physiol., 130, 1606-1613], the database has undergone extensive changes that are described in this publication. PMID- 16381967 TI - Phytome: a platform for plant comparative genomics. AB - Phytome is an online comparative genomics resource that can be applied to functional plant genomics, molecular breeding and evolutionary studies. It contains predicted protein sequences, protein family assignments, multiple sequence alignments, phylogenies and functional annotations for proteins from a large, phylogenetically diverse set of plant taxa. Phytome serves as a glue between disparate plant gene databases both by identifying the evolutionary relationships among orthologous and paralogous protein sequences from different species and by enabling cross-references between different versions of the same gene curated independently by different database groups. The web interface enables sophisticated queries on lineage-specific patterns of gene/protein family proliferation and loss. This rich dataset is serving as a platform for the unification of sequence-anchored comparative maps across taxonomic families of plants. The Phytome web interface can be accessed at the following URL: http://www.phytome.org. Batch homology searches and bulk downloads are available upon free registration. PMID- 16381968 TI - Plant MPSS databases: signature-based transcriptional resources for analyses of mRNA and small RNA. AB - MPSS (massively parallel signature sequencing) is a sequencing-based technology that uses a unique method to quantify gene expression level, generating millions of short sequence tags per library. We have created a series of databases for four species (Arabidopsis, rice, grape and Magnaporthe grisea, the rice blast fungus). Our MPSS databases measure the expression level of most genes under defined conditions and provide information about potentially novel transcripts (antisense transcripts, alternative splice isoforms and regulatory intergenic transcripts). A modified version of MPSS has been used to perform deep profiling of small RNAs from Arabidopsis, and we have recently adapted our database to display these data. Interpretation of the small RNA MPSS data is facilitated by the inclusion of extensive repeat data in our genome viewer. All the data and the tools introduced in this article are available at http://mpss.udel.edu. PMID- 16381969 TI - OryGenesDB: a database for rice reverse genetics. AB - Insertional mutant databases containing Flanking Sequence Tags (FSTs) are becoming key resources for plant functional genomics. We have developed OryGenesDB (http://orygenesdb.cirad.fr/), a database dedicated to rice reverse genetics. Insertion mutants of rice genes are catalogued by Flanking Sequence Tag (FST) information that can be readily accessed by this database. Our database presently contains 44166 FSTs generated by most of the rice insertional mutagenesis projects. The OryGenesDB genome browser is based on the powerful Generic Genome Browser (GGB) developed in the framework of the Generic Model Organism Project (GMOD). The main interface of our web site displays search and analysis interfaces to look for insertions in any candidate gene of interest. Several starting points can be used to exhaustively retrieve the insertions positions and associated genomic information using blast, keywords or gene name search. The toolbox integrated in our database also includes an 'anchoring' option that allows immediate mapping and visualization of up to 50 nucleic acid sequences in the rice Genome Browser of OryGenesDB. As a first step toward plant comparative genomics, we have linked the rice and Arabidopsis whole genome using all the predicted pairs of orthologs by best BLAST mutual hit (BBMH) connectors. PMID- 16381970 TI - DBD: a transcription factor prediction database. AB - Regulation of gene expression influences almost all biological processes in an organism; sequence-specific DNA-binding transcription factors are critical to this control. For most genomes, the repertoire of transcription factors is only partially known. Hitherto transcription factor identification has been largely based on genome annotation pipelines that use pairwise sequence comparisons, which detect only those factors similar to known genes, or on functional classification schemes that amalgamate many types of proteins into the category of 'transcription factor'. Using a novel transcription factor identification method, the DBD transcription factor database fills this void, providing genome wide transcription factor predictions for organisms from across the tree of life. The prediction method behind DBD identifies sequence-specific DNA-binding transcription factors through homology using profile hidden Markov models (HMMs) of domains. Thus, it is limited to factors that are homologus to those HMMs. The collection of HMMs is taken from two existing databases (Pfam and SUPERFAMILY), and is limited to models that exclusively detect transcription factors that specifically recognize DNA sequences. It does not include basal transcription factors or chromatin-associated proteins, for instance. Based on comparison with experimentally verified annotation, the prediction procedure is between 95% and 99% accurate. Between one quarter and one-half of our genome-wide predicted transcription factors represent previously uncharacterized proteins. The DBD (www.transcriptionfactor.org) consists of predicted transcription factor repertoires for 150 completely sequenced genomes, their domain assignments and the hand curated list of DNA-binding domain HMMs. Users can browse, search or download the predictions by genome, domain family or sequence identifier, view families of transcription factors based on domain architecture and receive predictions for a protein sequence. PMID- 16381971 TI - The Rice Annotation Project Database (RAP-DB): hub for Oryza sativa ssp. japonica genome information. AB - With the completion of the rice genome sequencing, a standardized annotation is necessary so that the information from the genome sequence can be fully utilized in understanding the biology of rice and other cereal crops. An annotation jamboree was held in Japan with the aim of annotating and manually curating all the genes in the rice genome. Here we present the Rice Annotation Project Database (RAP-DB), which has been developed to provide access to the annotation data. The RAP-DB has two different types of annotation viewers, BLAST and BLAT search, and other useful features. By connecting the annotations to other rice genomics data, such as full-length cDNAs and Tos17 mutant lines, the RAP-DB serves as a hub for rice genomics. All of the resources can be accessed through http://rapdb.lab.nig.ac.jp/. PMID- 16381972 TI - RMD: a rice mutant database for functional analysis of the rice genome. AB - Rice Mutant Database (RMD, http://rmd.ncpgr.cn) is an archive for collecting, managing and searching information of the T-DNA insertion mutants generated by an enhancer trap system. We have generated approximately 129 000 rice mutant (enhancer trap) lines that are now being gathered in the database. Information collected in RMD includes mutant phenotypes, reporter-gene expression patterns, flanking sequences of T-DNA insertional sites, seed availability and others, and can be searched by respective ID, keyword, nucleotide sequence or protein sequence on the website. This database is both a mutant collection for identifying novel genes and regulatory elements and a pattern line collection for ectopic expression of target gene in specific tissue or at specific growth stage. PMID- 16381973 TI - mtDB: Human Mitochondrial Genome Database, a resource for population genetics and medical sciences. AB - The mitochondrial genome, contained in the subcellular mitochondrial network, encodes a small number of peptides pivotal for cellular energy production. Mitochondrial genes are highly polymorphic and cataloguing existing variation is of interest for medical scientists involved in the identification of mutations causing mitochondrial dysfunction, as well as for population genetics studies. Human Mitochondrial Genome Database (mtDB) (http://www.genpat.uu.se/mtDB) has provided a comprehensive database of complete human mitochondrial genomes since early 2000. At this time, owing to an increase in the number of published complete human mitochondrial genome sequences, it became necessary to provide a web-based database of human whole genome and complete coding region sequences. As of August 2005 this database contains 2104 sequences (1544 complete genome and 560 coding region) available to download or search for specific polymorphisms. Of special interest to medical researchers and population geneticists evaluating specific positions is a complete list of (currently 3311) mitochondrial polymorphisms among these sequences. Recent expansions in the capabilities of mtDB include a haplotype search function and the ability to identify and download sequences carrying particular variants. PMID- 16381974 TI - Panzea: a database and resource for molecular and functional diversity in the maize genome. AB - Serving as a community resource, Panzea (http://www.panzea.org) is the bioinformatics arm of the Molecular and Functional Diversity in the Maize Genome project. Maize, a classical model for genetic studies, is an important crop species and also the most diverse crop species known. On average, two randomly chosen maize lines have one single-nucleotide polymorphism every approximately 100 bp; this divergence is roughly equivalent to the differences between humans and chimpanzees. This exceptional genotypic diversity underlies the phenotypic diversity maize needs to be cultivated in a wide range of environments. The Molecular and Functional Diversity in the Maize Genome project aims to understand how selection has shaped molecular diversity in maize and then relate molecular diversity to functional phenotypic variation. The project will screen 4000 loci for the signature of selection and create a wide range of maize and maize teosinte mapping populations. These populations will be genotyped and phenotyped, permitting high-power and high-resolution dissection of the traits and relating the molecular diversity to functional variation. Panzea provides access to the genotype, phenotype and polymorphism data produced by the project through user friendly web-based database searches and data retrieval/visualization tools, as well as a wide variety of information and services related to maize diversity. PMID- 16381975 TI - The Soybean Genome Database (SoyGD): a browser for display of duplicated, polyploid, regions and sequence tagged sites on the integrated physical and genetic maps of Glycine max. AB - Genomes that have been highly conserved following increases in ploidy (by duplication or hybridization) like Glycine max (soybean) present challenges during genome analysis. At http://soybeangenome.siu.edu the Soybean Genome Database (SoyGD) genome browser has, since 2002, integrated and served the publicly available soybean physical map, bacterial artificial chromosome (BAC) fingerprint database and genetic map associated genomic data. The browser shows both build 3 and build 4 contiguous sets of clones (contigs) of the soybean physical map. Build 4 consisted of 2854 contigs that encompassed 1.05 Gb and 404 high-quality DNA markers that anchored 742 contigs. Many DNA markers anchored sets of 2-8 different contigs. Each contig in the set represented a homologous region of related sequences. GBrowse was adapted to show sets of homologous contigs at all potential anchor points, spread laterally and prevented from overlapping. About 8064 minimum tiling path (MTP2) clones provided 13,473 BAC end sequences (BES) to decorate the physical map. Analyses of BES placed 2111 gene models, 40 marker anchors and 1053 new microsatellite markers on the map. Estimated sequence tag probes from 201 low-copy gene families located 613 paralogs. The genome browser portal showed each data type as a separate track. Tetraploid, octoploid, diploid and homologous regions are shown clearly in relation to an integrated genetic and physical map. PMID- 16381976 TI - Tomato Expression Database (TED): a suite of data presentation and analysis tools. AB - The Tomato Expression Database (TED) includes three integrated components. The Tomato Microarray Data Warehouse serves as a central repository for raw gene expression data derived from the public tomato cDNA microarray. In addition to expression data, TED stores experimental design and array information in compliance with the MIAME guidelines and provides web interfaces for researchers to retrieve data for their own analysis and use. The Tomato Microarray Expression Database contains normalized and processed microarray data for ten time points with nine pair-wise comparisons during fruit development and ripening in a normal tomato variety and nearly isogenic single gene mutants impacting fruit development and ripening. Finally, the Tomato Digital Expression Database contains raw and normalized digital expression (EST abundance) data derived from analysis of the complete public tomato EST collection containing >150,000 ESTs derived from 27 different non-normalized EST libraries. This last component also includes tools for the comparison of tomato and Arabidopsis digital expression data. A set of query interfaces and analysis, and visualization tools have been developed and incorporated into TED, which aid users in identifying and deciphering biologically important information from our datasets. TED can be accessed at http://ted.bti.cornell.edu. PMID- 16381977 TI - The TIGR Maize Database. AB - Maize is a staple crop of the grass family and also an excellent model for plant genetics. Owing to the large size and repetitiveness of its genome, we previously investigated two approaches to accelerate gene discovery and genome analysis in maize: methylation filtration and high C(0)t selection. These techniques allow the construction of gene-enriched genomic libraries by minimizing repeat sequences due to either their methylation status or their copy number, yielding a 7-fold enrichment in genic sequences relative to a random genomic library. Approximately 900,000 gene-enriched reads from maize were generated and clustered into Assembled Zea mays (AZM) sequences. Here we report the current AZM release, which consists of approximately 298 Mb representing 243,807 sequence assemblies and singletons. In order to provide a repository of publicly available maize genomic sequences, we have created the TIGR Maize Database (http://maize.tigr.org). In this resource, we have assembled and annotated the AZMs and used available sequenced markers to anchor AZMs to maize chromosomes. We have constructed a maize repeat database and generated draft sequence assemblies of 287 maize bacterial artificial chromosome (BAC) clone sequences, which we annotated along with 172 additional publicly available BAC clones. All sequences, assemblies and annotations are available at the project website via web interfaces and FTP downloads. PMID- 16381978 TI - Epitome: database of structure-inferred antigenic epitopes. AB - Immunoglobulin molecules specifically recognize particular areas on the surface of proteins. These areas are commonly dubbed B-cell epitopes. The identification of epitopes in proteins is important both for the design of experiments and vaccines. Additionally, the interactions between epitopes and antibodies have often served as a model for protein-protein interactions. One of the main obstacles in creating a database of antigen-antibody interactions is the difficulty in distinguishing between antigenic and non-antigenic interactions. Antigenic interactions involve specific recognition sites on the antibody's surface, while non-antigenic interactions are between a protein and any other site on the antibody. To solve this problem, we performed a comparative analysis of all protein-antibody complexes for which structures have been experimentally determined. Additionally, we developed a semi-automated tool that identified the antigenic interactions within the known antigen-antibody complex structures. We compiled those interactions into Epitome, a database of structure-inferred antigenic residues in proteins. Epitome consists of all known antigen/antibody complex structures, a detailed description of the residues that are involved in the interactions, and their sequence/structure environments. Interactions can be visualized using an interface to Jmol. The database is available at http://www.rostlab.org/services/epitome/. PMID- 16381979 TI - IMGT/LIGM-DB, the IMGT comprehensive database of immunoglobulin and T cell receptor nucleotide sequences. AB - IMGT/LIGM-DB is the IMGT comprehensive database of immunoglobulin (IG) and T cell receptor (TR) nucleotide sequences from human and other vertebrate species. It was created in 1989 by LIGM, Montpellier, France and is the oldest and the largest database of IMGT. IMGT/LIGM-DB includes all germline (non-rearranged) and rearranged IG and TR genomic DNA (gDNA) and complementary DNA (cDNA) sequences published in generalist databases. IMGT/LIGM-DB allows searches from the Web interface according to biological and immunogenetic criteria through five distinct modules depending on the user interest. For a given entry, nine types of display are available including the IMGT flat file, the translation of the coding regions and the analysis by the IMGT/V-QUEST tool. IMGT/LIGM-DB distributes expertly annotated sequences. The annotations hugely enhance the quality and the accuracy of the distributed detailed information. They include the sequence identification, the gene and allele classification, the constitutive and specific motif description, the codon and amino acid numbering, and the sequence obtaining information, according to the main concepts of IMGT-ONTOLOGY. They represent the main source of IG and TR gene and allele knowledge stored in IMGT/GENE-DB and in the IMGT reference directory. IMGT/LIGM-DB is freely available at http://imgt.cines.fr. PMID- 16381980 TI - EPD in its twentieth year: towards complete promoter coverage of selected model organisms. AB - The Eukaryotic Promoter Database (EPD) is an annotated non-redundant collection of eukaryotic POL II promoters, experimentally defined by a transcription start site (TSS). Access to promoter sequences is provided by pointers to positions in the corresponding genomes. Promoter evidence comes from conventional TSS mapping experiments for individual genes, or, starting from release 73, from mass genome annotation projects. Subsets of promoter sequences with customized 5' and 3' extensions can be downloaded from the EPD website. The focus of current development efforts is to reach complete promoter coverage for important model organisms as soon as possible. To speed up this process, a new class of preliminary promoter entries has been introduced as of release 83, which requires less stringent admission criteria. As part of a continuous integration process, new web-based interfaces have been developed, which allow joint analysis of promoter sequences with other bioinformatics resources developed by our group, in particular programs offered by the Signal Search Analysis Server, and gene expression data stored in the CleanEx database. EPD can be accessed at http://www.epd.isb-sib.ch. PMID- 16381981 TI - DBTSS: DataBase of Human Transcription Start Sites, progress report 2006. AB - DBTSS was first constructed in 2002 based on precise, experimentally determined 5' end clones. Several major updates and additions have been made since the last report. First, the number of human clones has drastically increased, going from 190,964 to 1,359,000. Second, information about potential alternative promoters is presented because the number of 5' end clones is now sufficient to determine several promoters for one gene. Namely, we defined putative promoter groups by clustering transcription start sites (TSSs) separated by <500 bases. A total of 8308 human genes and 4276 mouse genes were found to have putative multiple promoters. Third, DBTSS provides detailed sequence comparisons of user-specified TSSs. Finally, we have added TSS information for zebrafish, malaria and schyzon (a red algae model organism). DBTSS is accessible at http://dbtss.hgc.jp. PMID- 16381982 TI - HTPSELEX--a database of high-throughput SELEX libraries for transcription factor binding sites. AB - HTPSELEX is a public database providing access to primary and derived data from high-throughput SELEX experiments aimed at characterizing the binding specificity of transcription factors. The resource is primarily intended to serve computational biologists interested in building models of transcription factor binding sites from large sets of binding sequences. The guiding principle is to make available all information that is relevant for this purpose. For each experiment, we try to provide accurate information about the protein material used, details of the wet lab protocol, an archive of sequencing trace files, assembled clone sequences (concatemers) and complete sets of in vitro selected protein-binding tags. In addition, we offer in-house derived binding sites models. HTPSELEX also offers reasonably large SELEX libraries obtained with conventional low-throughput protocols. The FTP site contains the trace archives and database flatfiles. The web server offers user-friendly interfaces for viewing individual entries and quality-controlled download of SELEX sequence libraries according to a user-defined sequencing quality threshold. HTPSELEX is available from ftp://ftp.isrec.isb-sib.ch/pub/databases/htpselex/ and http://www.isrec.isb-sib.ch/htpselex. PMID- 16381983 TI - A new generation of JASPAR, the open-access repository for transcription factor binding site profiles. AB - JASPAR is the most complete open-access collection of transcription factor binding site (TFBS) matrices. In this new release, JASPAR grows into a meta database of collections of TFBS models derived by diverse approaches. We present JASPAR CORE--an expanded version of the original, non-redundant collection of annotated, high-quality matrix-based transcription factor binding profiles, JASPAR FAM--a collection of familial TFBS models and JASPAR phyloFACTS--a set of matrices computationally derived from statistically overrepresented, evolutionarily conserved regulatory region motifs from mammalian genomes. JASPAR phyloFACTS serves as a non-redundant extension to JASPAR CORE, enhancing the overall breadth of JASPAR for promoter sequence analysis. The new release of JASPAR is available at http://jaspar.genereg.net. PMID- 16381984 TI - MPromDb: an integrated resource for annotation and visualization of mammalian gene promoters and ChIP-chip experimental data. AB - We have developed Mammalian Promoter Database (MPromDb), a novel database that integrates gene promoters with experimentally supported annotation of transcription start sites, cis-regulatory elements, CpG islands and chromatin immunoprecipitation microarray (ChIP-chip) experimental results with intuitively designed presentation. Release 1.0 of MPromDb currently contains 36,407 promoters and first exons (19,170 from human, 15,953 from mouse and 1284 from rat), 3739 transcription factor (TF)-binding sites (2027 from human, 1181 mouse and 531 rat) and 224 TFs with links to PubMed and GenBank references. Target promoters of TFs that have been identified by ChIP-chip assay are integrated into the database. MPromDb serves as a portal for genome-wide promoter analysis of data generated by ChIP-chip experimental studies. MPromDb can be accessed from http://bioinformatics.med.ohio-state.edu/MPromDb. PMID- 16381985 TI - Androgen therapy in women. AB - Androgens in women either derive from direct ovarian production or from peripheral conversion of the adrenal sex steroid precursor, dehydroepiandrosterone, towards active androgens. Therefore, loss of adrenal or ovarian function, caused by Addison's disease or consequent to bilateral oophorectomy, results in severe androgen deficiency, clinically often associated with a loss of libido and energy. Importantly, physiological menopause does not necessarily lead to androgen deficiency, as androgen synthesis in the ovaries may persist despite the decline in estrogen production. However, the definition of female androgen deficiency, as recently provided by the Princeton consensus statement, is not precise enough and may lead to over-diagnosis due to the high prevalence of its diagnostic criteria: androgen levels below or within the lower quartile of the normal range and concurrent sexual dysfunction. Importantly, physiological menopause is not necessarily associated with androgen deficiency and therefore does not routinely require androgen therapy. Current replacement options include transdermal testosterone administration or dehydroepiandrosterone treatment, both of which have been shown to result in significant improvements, in particular in libido and mood, while effects on body composition and muscular function are not well documented. It is important to keep in mind that the number of randomized controlled trials is still limited and that currently none of the available preparations is officially approved for use in women. Currently, androgen replacement should be reserved for women with severe androgen deficiency due to an established cause and matching clinical signs and symptoms. PMID- 16381986 TI - Graves' disease and Hashimoto's thyroiditis in monozygotic twins: case study as well as transcriptomic and immunohistological analysis of thyroid tissues. AB - OBJECTIVE: To report on the rare simultaneous occurrence of Graves' disease (GD) and Hashimoto's thyroiditis (HT) in monozygotic twins. DESIGN: We compared the pattern of thyroid tissue-derived cDNAs to gain insight into previous and ongoing immune destruction and reconstruction processes using microarrays. The results were confirmed by immunohistology and real-time PCR. RESULTS: Destruction of thyroid tissue in HT reduced levels of thyrocyte-related cDNAs and cDNAs encoding extracellular matrix components, but increased levels of proteases involved in extracellular matrix degradation compared with GD. Lymphocytic infiltrates forming ectopic follicles replaced the thyroid tissue almost completely in HT. Thus, lymphocyte-related cDNA levels were higher in HT than in GD. The same was true for many chemokines and their receptors, which not only enable migration towards the thyroid but also maintain the lymphocytic infiltrate. HT also showed increased levels of cDNAs encoding molecules related to apoptosis than did GD. Surprisingly, the Th1- and Th2-specific cytokine profiles suggested for HT and GD respectively could not be confirmed. cDNAs encoding factors and receptors involved in angiogenesis were increased in GD compared with HT. CONCLUSIONS: Comparison of gene expression reflects the cellular differences between the two types of autoimmune thyroid disease in twins with identical genetic and similar environmental background. PMID- 16381987 TI - Epidemiology of subtypes of hypothyroidism in Denmark. AB - OBJECTIVE: Studies of hypothyroidism are often based on referred patients, and limited information is available on the incidence rates of subtypes of hypothyroidism in the general population. We therefore studied incidences of subtypes of primary, overt hypothyroidism in a Danish population cohort and compared incidences in two subcohorts with different levels of iodine intake. DESIGN: A prospective population-based study, monitoring a well-defined cohort representative of the Danish population. METHODS: The Danish Investigation of Iodine Intake and Thyroid Diseases registry of hyper- and hypothyroidism was established as part of the monitoring of the iodine fortification of salt in Denmark. A computer-based system linked to laboratory databases identified all patients diagnosed with new, biochemically overt hypothyroidism in populations living in Aalborg (moderate iodine deficiency, n = 311,102) and Copenhagen (mild iodine deficiency, n = 227,632). We subsequently evaluated all identified patients to verify incident thyroid disease, and subclassified hypothyroidism into nosological types. RESULTS: During a 4-year period (2,027,208 person-years) 685 new cases of overt hypothyroidism were diagnosed in the cohort; the incidence rate was 32.8 per 100,000 person-years (standardised to the Danish population). Nosological types of hypothyroidism were: spontaneous (presumably autoimmune) 84.4%, post-partum 4.7%, amiodarone-associated 4.0%, subacute thyroiditis 1.8%, previous radiation or surgery 1.8%, congenital 1.6% and lithium-associated 1.6%. Crude incidence rates were 29.0 around Aalborg and 40.6 in an area of Copenhagen. The higher incidence rate of hypothyroidism in the area with higher iodine intake was caused solely by more cases of spontaneous (presumably autoimmune) hypothyroidism, whereas the incidence of non-spontaneous hypothyroidism (all types combined) was significantly lower in the area with higher iodine intake. CONCLUSION: In a population-based study we observed a higher incidence of hypothyroidism with higher iodine intake. This was due solely to the entity of spontaneous hypothyroidism. The occurrence of overt hypothyroidism was relatively low in Denmark. PMID- 16381988 TI - The relative importance of genetic and environmental effects for the early stages of thyroid autoimmunity: a study of healthy Danish twins. AB - OBJECTIVE: In euthyroid individuals, autoantibodies to thyroid peroxidase (TPOab) and thyroglobulin (Tgab) are regarded as early markers of thyroid autoimmunity. Family and twin studies suggest that development of thyroid autoantibodies in first-degree relatives of patients with autoimmune thyroid disease is under genetic influence. We aimed to estimate the relative importance of genetic and environmental effects for the presence of thyroid autoantibodies in euthyroid subjects. METHODS: A representative sample of healthy twin pairs was identified through the Danish Twin Registry; 1372 individuals, divided into 283 monozygotic (MZ), 285 dizygotic same sex (DZ), and 118 opposite sex twin pairs were investigated. Serum TPOab and serum Tgab were measured. Proband-wise concordance and intraclass correlations were calculated, and quantitative genetic modelling was performed. RESULTS: Probandwise concordance and intraclass correlations were consistently higher for MZ than for DZ twin pairs indicating genetic influence. Genetic components (with 95% confidence intervals) accounted for 73% (46-89%) of the liability of being thyroid antibody positive. Adjusting for covariates (age, TSH and others), the estimate for genetic influence on serum TPOab concentrations was 61% (49-70%) in males and 72% (64-79%) in females. For serum Tgab concentrations, the estimates were 39% (24-51%) and 75% (66-81%) respectively. CONCLUSIONS: Early markers of thyroid autoimmunity appear to be under strong genetic influence. The analyses suggest that it is the same set of genes that operate in males and females. However, complex mechanisms such as dominance and/or epistasis may be involved. PMID- 16381989 TI - Lack of association between thyroid autoantibodies and parity in a population study argues against microchimerism as a trigger of thyroid autoimmunity. AB - BACKGROUND: Thyroid autoimmunity is more common in females than in males. One possible explanation for this female preponderance may be the effect of oestrogens on the immune system. It has also been suggested that foetal microchimerism involving transfer of foetal cells into maternal tissue during pregnancy may play an important role. OBJECTIVE: We investigated the association between the presence of circulating thyroid autoantibodies and previous pregnancy, parity and the use of oral contraceptives (OCs) and hormone replacement therapy (HRT) in a population cohort. METHODS: We examined 3712 women randomly selected from the general population. Serum was analysed for thyroid peroxidase antibody (TPO-Ab) and thyroglobulin antibody (Tg-Ab) using assays based on an RIA technique (DYNO test). Data were analysed in logistic regression models to adjust for possible confounders. Women previously treated for thyroid disease or with pregnancy within 1 year prior to the study were excluded from the analyses. RESULTS: In both univariate and multivariate models and whether the presence of TPO-Ab and Tg-Ab was investigated alone or in combination, findings were negative with respect to an association between circulating thyroid antibodies and previous pregnancy, number of pregnancies, parity and previous abortion. There was no association between thyroid autoantibodies and use of OCs. Women aged 60-65 years receiving HRT now or previously had a lower prevalence of Tg-Ab (univariate, P = 0.01; multivariate, P = 0.02). No such association was observed between HRT and TPO-Ab. CONCLUSION: In this population study there was no association between previous pregnancy, parity and thyroid antibodies, which argues against the role of microchimerism as a trigger of thyroid autoimmunity. Exogenous oestrogens may reduce aspects of autoimmunity. PMID- 16381990 TI - Inflammation is a modulator of the insulin-like growth factor (IGF)/IGF-binding protein system inducing reduced bioactivity of IGFs in cystic fibrosis. AB - OBJECTIVE: In inflammatory bowel diseases, increased serum interleukin (IL)-6 levels are associated with high serum insulin-like growth factor-binding protein 2 (IGFBP-2) levels, and cytokines modify the insulin-like growth factor (IGF)/IGFBP system in models in vitro. In cystic fibrosis (CF) the IGF/IGFBP system has not been extensively studied, and relationships with proinflammatory cytokines have not been explored. The aim of this study was to investigate the IGF/IGFBP system and verify changes dependent on IL-1beta, IL-6, tumour necrosis factor alpha (TNFalpha), and insulin. METHODS: Eighteen subjects with CF (mean age 26.6 +/- 1.1 years) and 18 controls, comparable for age, sex, and body mass index, were enrolled. Serum IGF-I, IGF-II, IGFBP-2, IGFBP-3, IL-1beta, IL-6, TNFalpha, insulin and C-peptide were measured. Different molecular forms of IGFBP 2 and IGFBP-3 were investigated by Western immunoblotting. The patients were analysed as a whole and as two subgroups depending on established clinical criteria (Swachman-Kulczycki score). RESULTS: Patients had higher serum concentrations of IL-1beta, IL-6, TNFalpha and IGFBP-2 than controls. Serum concentrations of IGF-I and IGF-II were significantly lower and insulin and C peptide levels significantly increased in CF compared with healthy controls whereas IGFBP-3 serum concentrations were similar, with comparable IGF-I/IGFBP-3 and decreased IGF-I/IGFBP-2 and IGF-II/IGFBP-2 molar ratios. From correlation analysis we detected a significant positive correlation between IGFBP-2 and IL-6 and a negative correlation between IGFBP-2 and IGFBP-3. CONCLUSIONS: Our findings suggest that inflammation is an important modulator of the IGF/IGFBP system with an overall reduction in IGF bioactivity in CF. PMID- 16381991 TI - Effects of gastric bypass on the GH/IGF-I axis in severe obesity--and a comparison with GH deficiency. AB - OBJECTIVE: Overfeeding suppresses GH secretion and makes evaluation of a suspected GH deficiency (GHD) difficult. In normal weight subjects, gender is known to influence GH concentrations, which is most apparent in the ambulatory, morning-fasted state. In this study, we examined the GH/IGF-I axis in obese men and women and the effect of surgically induced weight loss. DESIGN: Sixty-three subjects (body mass index (BMI) 45 +/- 6 kg/m2; 54 women, 9 men) were studied prior to, and 6 and 12 months following Roux-en-Y gastric bypass (RYGBP) surgery. Fifty-four patients with classic GHD (BMI 27 +/- 6 kg/m2; 35 men, 19 women) were included for comparison. METHODS: Hormones were analysed in fasting morning serum samples. RESULTS: RYGBP resulted in a decreased BMI to 35 +/- kg/m2 at 6 months and 32 +/- 6 kg/m2 at 12 months. GH and IGF-I increased at 6 months in the women and at 12 months in both sexes by > or = 300 and 11% respectively. Prior to RYGBP, GH concentrations were low in the obese men and similar to those of GHD men (mean 0.09 mU/l). Obese women had tenfold higher values than obese men and sevenfold higher than GHD women. IGF-I levels were in the low reference range in the obese and below -2 S.D. for age in 13%. CONCLUSIONS: Surgically induced weight loss partially restores GH secretion. Despite a marked suppression of GH values, a gender influence is maintained in severe obesity. In obese women, single morning GH and IGF-I values seem sufficient to exclude a suspicion of classic GHD. PMID- 16381992 TI - Ghrelin, insulin sensitivity and postprandial glucose disposal in overweight and obese children. AB - OBJECTIVE: To explore the changes of ghrelin circulating levels induced by a mixed meal and their relationship with postprandial substrate oxidation rates in overweight and obese children with different levels of insulin sensitivity. METHODS: A group of ten boys (age 9-12 years) with different levels of overweight (standard deviation score of body mass index: 1.6-3.2) was recruited. Body composition was measured by dual-energy X-ray absorptiometry. Insulin sensitivity was assessed by a frequently sampled i.v. glucose tolerance test. Pre-prandial and postprandial (3 h) substrate oxidation was measured by indirect calorimetry. The energy content of the test meal (16% protein, 36% carbohydrate and 48% fat) was 40% of pre-prandial energy expenditure (kJ/day). RESULTS: Pre-prandial serum concentration of total ghrelin was 701.4+/-66.9 pg/ml (S.E.M.). The test meal induced a rapid decrease in ghrelin levels and maximal decrease was 27.3+/-2.7% below baseline. Meal intake induced a progressive increase of the carbohydrate oxidation rate for 45 min after food ingestion, followed by a slow decrease without returning to pre-prandial values. Postprandial cumulative carbohydrate oxidation was 16.9+/-0.8 g/3 h. Insulin sensitivity and postprandial maximal decrease of ghrelin concentration showed a significant correlation (r = 0.803, P < 0.01). Moreover, the postprandial carbohydrate oxidation rate correlated with the area under the curve for both insulin (r = 0.673, P < 0.03) and ghrelin (r = 0.661, P < 0.04). CONCLUSIONS: A relevant association between postprandial insulin-mediated glucose metabolism and ghrelin secretion in children with different levels of overweight was found. It is possible that the maintenance of an adequate level of insulin sensitivity and glucose oxidation may affect appetite regulation by favoring a more efficient postprandial ghrelin reduction. PMID- 16381993 TI - GH effect on enzyme activity of 11betaHSD in abdominal obesity is dependent on treatment duration. AB - OBJECTIVE: In the past years the interaction of GH and 11beta hydroxysteroid dehydrogenase (11betaHSD) in the pathogenesis of central obesity has been suggested. DESIGN: We studied the effects of 9 months of GH treatment on 11betaHSD activity and its relationship with body composition and insulin sensitivity in 30 men with abdominal obesity, aged 48-66 years, in a randomised, double-blind, placebo-controlled trial. METHODS: Urinary steroid profile was used to estimate 11betaHSD type 1 and 2 (11betaHSD1 and 11betaHSD2) activities. Abdominal s.c. and visceral adipose tissues were measured using computed tomography. Glucose disposal rate (GDR) obtained during a euglycaemic hyperinsulinaemic glucose clamp was used to assess insulin sensitivity. RESULTS: In the GH-treated group the 11betaHSD1 activity decreased transiently after 6 weeks (P < 0.01) whereas 11betaHSD2 increased after 9 months of treatment (P < 0.05). Between 6 weeks and 9 months, GDR increased and visceral fat mass decreased. Changes in 11betaHSD1 correlated with changes in visceral fat mass between baseline and 6 weeks. There were no significant correlations between 11betaHSD1 and 11betaHSD 2 and changes in GDR. DISCUSSION: The study demonstrates that short- and long-term GH treatment has different effects on 11betaHSD1 and 11betaHSD2 activity. Moreover, the data do not support that long-term metabolic effects of GH are mediated through its action on 11betaHSD. PMID- 16381994 TI - Plasma osteoprotegerin levels are associated with glycaemic status, systolic blood pressure, kidney function and cardiovascular morbidity in type 1 diabetic patients. AB - OBJECTIVE: The bone-related peptide osteoprotegerin (OPG) has recently been found in increased amounts in the vasculature in diabetes. It is produced by vascular smooth muscle and endothelial cells, and may be implicated in the development of vascular calcifications. OPG is present in the circulation, where increased amounts have been observed in patients with diabetes. In this study, we examined whether plasma OPG is associated with the glycaemic and vascular status of patients with type 1 diabetes. METHODS: Two gender-, age- and duration-comparable groups of type 1 diabetic patients either with (n = 199) or without (n = 192) signs of diabetic nephropathy were studied. Plasma OPG was determined by an ELISA. RESULTS: The plasma OPG concentration was significantly higher in patients with nephropathy than those without (3.11 (2.49-3.99) vs 2.57 (2.19-3.21) (median (interquartiles), ng/ml), P < 0.001). Plasma OPG correlated with haemoglobin A1c (HbA1c), systolic blood pressure and age in both groups and, in addition, with kidney function in the nephropathic group. These correlations remained significant in multivariate models. In addition, we found that plasma OPG concentrations were increased among patients with cardiovascular diseases (CVD), both in the normoalbuminuric and the nephropathic groups. The differences between nephropathic and normoalbuminuric, as well as subgroups with and without CVD, could largely be ascribed to changes in HbA1c, age, systolic blood pressure and creatinine. CONCLUSION: OPG is associated with glycaemic control and CVD in patients with type 1 diabetes, compatible with the hypothesis that OPG is associated with the development of diabetic vascular complications. PMID- 16381995 TI - Effects of retinoid therapy on insulin sensitivity, lipid profile and circulating adipocytokines. AB - OBJECTIVE: In vitro and in vivo models indicate that all-trans retinoic acids influence glucose and lipid metabolism. We aimed to evaluate the effects of chronic treatment with acitretin, an all-trans retinoic acid, on glucose metabolism, lipid profile and adiponectin and resistin levels. DESIGN: Ten normoglycemic, normolipemic patients affected with psoriasis vulgaris were studied before and after 1 and 3 months of oral treatment with 35 microg of acitretin. METHODS: Glucose metabolism, lipid profile, and adiponectin and resistin levels were evaluated in basal conditions and after acitretin treatment. Ten healthy subjects matched for age, body mass index (BMI) and insulin sensitivity were studied as controls. RESULTS: One-month acitretin treatment reduced psoriasis activity, insulin sensitivity, evaluated as QUICKI values (0.364 +/- 0.034 versus 0.329 +/- 0.051; P < 0.05) and HOMA-IR index (1.53 +/- 0.73 versus 2.59 +/- 1.41; P < 0.05), and high-density lipoprotein (HDL) cholesterol levels (45.2 +/- 11.7 versus 39.4 +/- 10.4 mg/dl; P = 0.01). The impairment in glucose and lipid homeostasis was transient and not associated to BMI variations. Adiponectin levels did not change during the treatment, while resistin levels, which were higher in untreated patients than in controls (9.4 +/ 4.4 versus 6.2 +/- 2.1 ng/ml; P = 0.05), fell within the normal range after 1 and 3 months of therapy. The normalization of resistin levels occurred without significant changes in circulating tumor necrosis factor alpha (TNFalpha) levels, which persisted elevated throughout the treatment. CONCLUSIONS: Treatment with a low dose of acitretin induced a mild, transient reduction of insulin sensitivity and HDL-cholesterol levels that was not related to modifications of adiponectin, resistin and TNFalpha levels. Although the role of resistin in humans remains elusive, the levels of this adipocytokine seem to be affected, at least in part, by retinoids. PMID- 16381996 TI - Adiponectin and the development of diabetes in patients with coronary artery disease and impaired fasting glucose. AB - OBJECTIVES: Adiponectin has insulin-sensitizing properties, and high adiponectin levels have been shown to be associated with reduced risk of developing diabetes. Patients with coronary artery disease (CAD) have relatively low adiponectin levels and high prevalence of glucose intolerance. The role of adiponectin in predicting the development of diabetes in this high-risk group has not been determined. The study aimed to determine whether baseline adiponectin levels predict the development of diabetes in a group of patients with CAD and impaired fasting glucose (IFG). METHODS: A total of 588 patients who participated in the Bezafibrate Infarction Prevention (BIP) study and who had at baseline fasting glucose of 100-125 mg/dl were included and followed for 6.2+/-1.3 years. Adiponectin was determined in frozen plasma samples taken at baseline. RESULTS: Of the patients with IFG at baseline, 256 (44%) developed diabetes during follow up. The patients who developed diabetes had at baseline higher body-mass index, fasting glucose, C-reactive protein, triglycerides, homeostatic assessment of insulin resistance (HOMA-IR) and diastolic blood pressure than patients who did not develop diabetes. Adiponectin levels at baseline were significantly lower in patients who developed diabetes than in patients who did not develop diabetes (P = 0.009, nonparametric Kruskall-Wallis test). An increase of 1 unit of natural logarithm of adiponectin level was associated with a hazard ratio of 0.77 (95% CI, 0.61-0.96) for diabetes development. CONCLUSION: Patients with CAD and IFG have a very high rate of conversion to type 2 diabetes. Even in this high-risk group, high adiponectin levels are associated with reduced risk of developing diabetes. PMID- 16381997 TI - Regulation of plasma fibroblast growth factor 23 by calcium in primary hyperparathyroidism. AB - OBJECTIVE: While the importance of fibroblast growth factor (FGF)-23 is established in phosphate-wasting disorders, little is known about the mechanisms regulating its circulating level. To investigate the role of parathyroid hormone (PTH) and calcium in FGF-23 metabolism, we examined plasma FGF-23 levels in patients with primary hyperparathyroidism (PHPT). PATIENTS AND METHODS: Fifty patients with PHPT and 52 controls were employed in this study. Plasma was obtained from 18 PHPT patients who underwent parathyroidectomy (PTX) on the first postoperative morning without vitamin D administration. Time-course samples were also obtained from 5 of 18 PTX patients without vitamin D analogs or calcium administration. The expression of Fgf23 on resected parathyroid glands was analyzed by reverse transcription (RT)-PCR and immunohistochemistry. RESULTS: FGF 23 was significantly elevated in PHPT patients compared with controls. FGF-23 levels were significantly correlated positively with serum corrected calcium and intact PTH levels, and negatively with creatinine clearance and inorganic phosphate, among which creatinine clearance and corrected calcium were independently associated factors. In 18 PTX patients, postoperative FGF-23 levels were significantly decreased compared with preoperative levels. Corrected-calcium levels were significantly decreased 1 h after PTX, and this was followed by a reduction in plasma FGF-23 levels in time-course study. In addition, postoperative FGF-23 levels in 18 PTX patients were significantly correlated with corrected calcium, consistent with a role of serum calcium as one of the major regulators of FGF-23. The absence of Fgf23 expression in parathyroid glands indicated that the parathyroid glands were not major sources of circulating FGF 23. CONCLUSIONS: Serum calcium may regulate circulating FGF-23 levels in PHPT. PMID- 16381998 TI - Hormone replacement therapy and mortality in 52- to 70-year-old women: the Kuopio Osteoporosis Risk Factor and Prevention Study. AB - OBJECTIVES: To analyze prospectively the association between hormone replacement therapy (HRT) and mortality in women before old age. DESIGN AND METHODS: A group of 11,667 women (91% of the age cohort of the area) aged 52-62 years from the population-based Kuopio Osteoporosis Risk Factor and Prevention Study were followed for 7 years in 1994-2001. Information about HRT use and health events was obtained from two repeated questionnaires in 1989 and 1994. Information about deaths and causes of death from the follow-up period was obtained from the Statistics Finland. Cox's proportional-hazards models were used to calculate risk of death related to the use of HRT. RESULTS: At the start of follow-up, 2203 women had used HRT > 5 years, 3945 women < or = 5 years and 5519 women had never used it. During the follow-up, 361 deaths occurred. Compared with non-users of HRT, the adjusted hazard ratio (HR) of death from any cause was 1.05 (95% confidence interval (CI) 0.80-1.36) in women who used HRT < or = 5 years and 1.06 (95% CI 0.78-1.46) in women who used HRT > 5 years. The adjusted HR for coronary heart disease (CHD) mortality in women who used HRT < or = 5 years was 0.79 (95% CI 0.36-1.73), and in women who used HRT > 5 years, 2.16 (95% CI 0.93-4.98). For breast cancer mortality the adjusted HR for < or = 5 years of HRT use was 0.96 (95% CI 0.32-2.82) and 2.62 (95% CI 0.98-7.00) for > 5 years of HRT use. CONCLUSIONS: History of HRT use does not affect overall or CHD mortality in women. More than 5 years of HRT use may increase the risk of breast cancer mortality. PMID- 16381999 TI - Restoration of the coupling process and normalization of bone mass following successful treatment of endogenous Cushing's syndrome: a prospective, long-term study. AB - OBJECTIVE: Endogenous Cushing's syndrome (CS) is associated with bone loss and an increased risk of fractures. However, the long-term outcome of treatment on bone health has not been adequately clarified. DESIGN: We followed 33 patients with active CS prospectively before and twice after treatment (mean follow-up 33 (n = 25) and 71 months (n = 18), respectively). The patients were compared to age-, sex- and body mass index (BMI)-matched controls, also followed longitudinally. METHODS: Bone mineral indices (bone mineral density (BMD), bone mineral content (BMC) and bone area) were evaluated in the lumbar spine (LS), femoral neck (FN), and total body (TB) by dual-energy X-ray absorptiometry (DXA). Biochemical markers of bone turnover were assessed by serum levels of osteocalcin and C terminal telopeptides of Type-1 collagen (CTX-1). RESULTS: Mann-Whitney rank sum tests showed that BMD of the LS, FN and TB was reduced by 14.8% (P < 0.001), 15.7% (P < 0.001), and 9.2% (P < 0.001) in CS vs. controls at baseline, with markedly reduced serum osteocalcin (P = 0.014) and increased CTX-1 (P = 0.012) levels, but no correlation between markers. At first follow-up, BMD was increased in LS (7.9%, P < 0.001) and FN (3.5%, P = 0.003) compared to baseline. The time dependent rise in BMD (LS (r = 0.59; P = 0.002) and FN (r = 0.52; P = 0.007); Spearman's rank correlation), in CS was paralleled by increased osteocalcin (275%, P < 0.001) and correlation between biochemical markers (r = 0.92, P < 0.001; Pearson's correlation). TB BMD did not increase significantly before the second follow-up, when BMD Z-scores were normalized in all three compartments. CONCLUSION: Our observations demonstrate restoration of coupled bone remodeling and normalization of bone mineral density in all measured skeletal compartments of treated CS patients after prolonged recovery, first significant in predominantly trabecular bone (i.e. lumbar spine). PMID- 16382000 TI - Three-month sustained-release triptorelin (11.25 mg) in the treatment of central precocious puberty. AB - OBJECTIVE: Depot GnRH agonists are commonly used in the treatment of central precocious puberty (CPP). The triptorelin 11.25 mg 3-month depot, currently used in adult indications, had not previously been evaluated in CPP. DESIGN: This was a multicenter, open-label, 12 month trial conducted in 64 CPP children (54 girls and 10 boys), treated quarterly. METHODS: Children with a clinical onset of pubertal development before the age of 8 years (girls) or 9 years (boys), pubertal response of LH to GnRH > or = 7 IU/l, advanced bone age > 1 year, enlarged uterus (> or = 36 mm) and testosterone level > or = 0.5 ng/ml (boys), were included. Suppression of gonadotropic activation, as determined from serum LH, FSH, estradiol or testosterone, and pubertal signs were assessed at Months 3, 6 and 12. RESULTS: GnRH-stimulated peak LH < or = 3 IU/l, the main efficacy criterion, was met in 53 out of 62 (85%), 60 out of 62 (97%) and 56 out of 59 (95%) of the children at Months 3, 6 and 12 respectively. Serum FSH and sex steroids were also significantly reduced, while pubertal development regressed in most patients. Mean residual triptorelin levels were stable from Month 3 through to Month 12. The triptorelin 3-month depot was well tolerated. Severe injection pain was experienced in only one instance. Five girls experienced mild-to moderate or severe (one girl) withdrawal bleeding. CONCLUSIONS: The triptorelin 3 month depot efficiently suppresses the pituitary-gonadal axis and pubertal development in children with CPP. This formulation allows a 3-fold reduction, over the once-a-month depot, in the number of i.m. injections required each year. PMID- 16382001 TI - Postnatal penile length and growth rate correlate to serum testosterone levels: a longitudinal study of 1962 normal boys. AB - OBJECTIVE: Infant boys show a brief activation of their hypothalamic-pituitary gonadal axis shortly after birth, the physiological significance of which is poorly understood. The objective of the study was to investigate the correlation between endogenous testosterone levels and penile size and growth. DESIGN: Prospective, longitudinal population-based study taking place at two large primary obstetric centres at the University Hospitals of Copenhagen, Denmark, and Turku, Finland. METHODS: Infant boys, 728 Danish and 1234 Finnish, underwent clinical examinations at 0, 3, 18 and 36 months in Denmark and at 0, 3 and 18 months in Finland with blood samples taken at 3 months (n = 630). Penile length and growth were registered and reproductive hormones (testosterone, sex hormone binding globulin, oestradiol) were analysed. RESULTS: Penile length increased from birth (3.49+/-0.4 cm) to 3 years of age (4.53+/-0.51 cm) with the highest growth velocity from birth to 3 months (1.0 mm/month). Penile length and growth were significantly, positively correlated to serum testosterone (r = 0.31 and 0.076, P = 0.006 and 0.001 respectively) and to free testosterone index (r = 0.385 and 0.094, P = 0.0001 and 0.0001 respectively). CONCLUSIONS: We found that endogenous testosterone was significantly associated with penile size and growth rate in infant boys. Thus, the postnatal surge in reproductive hormones appears to be important for genital growth. Our data may serve as an updated reference for normal penile length in Caucasian boys up to 3 years of age. PMID- 16382002 TI - Influence of methyltestosterone postmenopausal therapy on plasma lipids, inflammatory factors, glucose metabolism and visceral fat: a randomized study. AB - BACKGROUND: There has been a growing interest in treating postmenopausal women with androgens. However, hyperandrogenemia in females has been associated with increased risk of cardiovascular disease. OBJECTIVE: We aimed to assess the effects of androgen replacement on cardiovascular risk factors. DESIGN: Thirty seven postmenopausal women aged 42-62 years that had undergone hysterectomy were prospectively enrolled in a double-blind protocol to receive, for 12 months, percutaneous estradiol (E2) (1 mg/day) combined with either methyltestosterone (MT) (1.25 mg/day) or placebo. METHODS: Along with treatment, we evaluated serum E2, testosterone, sex hormone-binding globulin (SHBG), free androgen index, lipids, fibrinogen, and C-reactive protein; glucose tolerance; insulin resistance; blood pressure; body-mass index; and visceral and subcutaneous abdominal fat mass as assessed by computed tomography. RESULTS: A significant reduction in SHBG (P < 0.001) and increase in free testosterone index (P < 0.05; Repeated measures analysis of variance) were seen in the MT group. Total cholesterol, triglycerides, fibrinogen, and systolic and diastolic blood pressure were significantly lowered to a similar extent by both regimens, but high-density lipoprotein cholesterol decreased only in the androgen group. MT-treated women showed a modest rise in body weight and gained visceral fat mass relative to the other group (P < 0.05), but there were no significant detrimental effects on fasting insulin levels and insulin resistance. CONCLUSION: This study suggests that the combination of low-dose oral MT and percutaneous E2, for 1 year, does not result in expressive increase of cardiovascular risk factors. This regimen can be recommended for symptomatic postmenopausal women, although it seems prudent to perform baseline and follow-up lipid profile and assessment of body composition, especially in those at high risk of cardiovascular disease. PMID- 16382003 TI - Metabolic syndrome in polycystic ovary syndrome (PCOS): lower prevalence in southern Italy than in the USA and the influence of criteria for the diagnosis of PCOS. AB - OBJECTIVE: Metabolic syndrome (MBS) is a common disorder and is thought to be extremely prevalent in polycystic ovary syndrome (PCOS). In the USA the prevalence of MBS in PCOS has been reported to be as high as 43-46% using Adult Treatment Panel III (ATP-III) criteria. Because of differences in diet, lifestyle and genetic factors, we postulated that the prevalence of MBS might not be as high in Italian women. This study sought to determine the prevalence of MBS in Italian women using both the ATP-III and the World Health Organization (WHO) criteria and to determine whether the prevalence is influenced by the way in which PCOS is diagnosed. DESIGN: Assessment of the prevalence of MBS in 282 women with PCOS, aged 18-40 years, living in western Sicily. Eighty-five age- and weight-matched normal women served as controls. METHODS: Patients were divided into those with chronic anovulation and hyperandrogenism (classic PCOS; n = 225) and others with hyperandrogenism and polycystic ovaries but who were ovulatory (ovulatory PCOS; n = 57). A 75 g oral glucose tolerance test was carried out, as were lipid determinations; insulin resistance was assessed by the Quantitative Insulin-Sensitivity Check Index (QUICKI). We used ATP-III and WHO criteria to diagnose MBS. RESULTS: Using ATP-III criteria, the prevalence of MBS was 8.2% and using WHO criteria it was 16% in Italian women with PCOS. In controls, the prevalence was 2.4% using both methods. In classic PCOS patients, MBS was higher (8.9% by ATP-III, 17.3% by WHO) than in ovulatory PCOS (5% and 10.6% respectively). Body weight significantly modified prevalence rates. CONCLUSION: MBS is substantially higher in women with PCOS than in the general population, and the prevalence is higher in those women diagnosed by classic criteria. However, the prevalence of MBS in PCOS appears to be much lower in Italy than in the USA. PMID- 16382004 TI - Evaluation of pituitary function in the fatigued patient: a review of 59 cases. AB - OBJECTIVE: The aim of this study was to review the results of dynamic pituitary testing in patients presenting with fatigue. METHODS: We reviewed clinical histories and insulin tolerance test (ITT) results of 59 patients who presented with fatigue and other symptoms of glucocorticoid insufficiency over a 4-year period. All patients referred for ITT had an early-morning cortisol level of <400 nM and a low or normal ACTH level. RESULTS: Peak cortisol and GH responses following insulin-induced hypoglycaemia were normal in only seven patients (12%). Median age of the remaining 52 patients was 47 years (range, 17-67 years); all but five were female. Common presenting symptoms were neuroglycopaenia (n = 47), depression (n = 37), arthralgia and myalgia (n = 28), weight gain (n = 25), weight loss (n = 9), postural dizziness (n = 15) and headaches (n = 13). Other medical history included autoimmune disease (n = 20; particularly Hashimoto's thyroiditis, Graves' disease and coeliac disease), postpartum (n = 8) and gastrointestinal (n = 2) haemorrhage and hyperprolactinaemia (n = 13). 31 subjects had peak cortisol levels of <500 nM (suggestive of ACTH deficiency; 18 of whom had levels < 400 nM) and a further six had indeterminate results (500-550 nM). The remaining 15 subjects had normal cortisol responses (median 654 nM; range, 553-1062 nM) but had low GH levels following hypoglycaemic stimulation (5.9 mU/l; 3-11.6 mU/l). CONCLUSION: Our results suggest that patients presenting with fatigue and symptoms suggestive of hypocortisolism should be considered for screening for secondary adrenal insufficiency, particularly in the presence of autoimmune disease or a history of postpartum or gastrointestinal haemorrhage. Whether physiological glucocorticoid replacement improves symptoms in this patient group is yet to be established. PMID- 16382005 TI - RT-PCR analysis of corticotroph-associated genes expression in carcinoid tumours in the ectopic-ACTH syndrome. AB - OBJECTIVE: ACTH is frequently produced in non-pituitary tumours, leading to the ectopic-ACTH syndrome, but the molecular mechanisms of its expression remain obscure. This study was aimed at understanding the transcription mechanisms of the ACTH-precursor gene in carcinoid tumours of the lung or thymus. DESIGN: Transcripts coding for a series of corticotroph-associated transcription factor genes were detected, together with markers of the corticotroph phenotype. We studied a series of 41 carcinoid tumours including 15 with proven ectopic-ACTH syndrome. METHODS: Specific RT-PCR reactions were designed for each gene including alternatively spliced isoforms. RESULTS: The markers of the corticotroph phenotype were detected in all ACTH-positive tumours. Expression of the Tpit and Pitx1 genes were not restricted to ACTH-positive tumours but were also detected in many ACTH-negative carcinoids. Only a subset of ACTH-negative tumours expressed NAK-1/Nur77, and NeuroD1 expression was detected in approximately 50% of the tumours regardless of their secretory status. The glucocorticoid receptor alpha was detected in every tumour in contrast to its beta isoform detectable in a few tumours only. Chicken ovalbumin upstream promoter-transcription factor 1 (COUP-TF1) and peroxisome proliferator-activated receptor (PPAR) gamma2 were expressed in 50% of the tumours of each group whereas PPARgamma1 was expressed in almost every tumour. CONCLUSIONS: ACTH-positive carcinoids do not share a characteristic expression pattern of the corticotroph associated transcription factor genes, suggesting that the transcriptional mechanisms of the ACTH-precursor gene differ from those in normal pituitary corticotrophs. Expression of Tpit and Pitx1 genes in most carcinoids suggests that some aspects of the pituitary corticotroph phenotype may belong to general carcinoid differentiation. PMID- 16382006 TI - Osteochondral autograft transplantation for osteochondritis dissecans of the elbow in juvenile baseball players: minimum 2-year follow-up. AB - BACKGROUND: Osteochondral autografts have recently become popular to treat articular cartilage defects, and they are used for unstable osteochondritis dissecans lesions as a means of biological fixation. PURPOSE: To evaluate the clinical results of osteochondral autograft transfer for osteochondritis dissecans of the elbow. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Osteochondral autograft transfer was performed on 18 baseball players (mean age, 13.6 years) with osteochondritis dissecans of the elbow. These included 9 lesions that were grade 3 (separated but in situ) and 9 lesions that were grade 4 (displaced fragment with osteochondral defect) based on magnetic resonance imaging. All patients were evaluated with a scoring system, radiographs, and magnetic resonance imaging, with the mean follow-up at 3.5 years. RESULTS: In patients with grade 3 lesions, the subjective score was increased, but the objective score did not change. Six of 9 patients returned to their previous sports performance levels. One quit baseball because of academic reasons, 1 changed his position, and 1 changed to softball. In patients with grade 4 lesions, both subjective and objective scores were increased significantly. All but 1 patient returned to their previous sports performance levels. In the 3 grade 4 lesions with a wide osteochondral defect, the irregularity of the articular surface remained on magnetic resonance imaging. CONCLUSION: Osteochondral autograft transplantation is a useful treatment for reattachment of the lesion as well as osteochondral resurfacing of elbow osteochondritis dissecans. PMID- 16382007 TI - Anterior cruciate ligament injuries in female athletes: Part 2, a meta-analysis of neuromuscular interventions aimed at injury prevention. AB - Female athletes have a 4 to 6 times higher incidence of anterior cruciate ligament injury than do male athletes participating in the same landing and pivoting sports. This greater risk of anterior cruciate ligament injury, coupled with a geometric increase in participation (doubling each decade), has led to a significant rise in anterior cruciate ligament injuries in female athletes. The gender gap in anterior cruciate ligament injury, combined with evidence that the underpinnings of this serious health problem are neuromuscular in nature, leads to the development of neuromuscular interventions designed to prevent injury. A systematic review of the published literature yielded 6 published interventions targeted toward anterior cruciate ligament injury prevention in female athletes. Four of 6 significantly reduced knee injury incidence, and 3 of 6 significantly reduced anterior cruciate ligament injury incidence in female athletes. A meta analysis of these 6 studies demonstrates a significant effect of neuromuscular training programs on anterior cruciate ligament injury incidence in female athletes (test for overall effect, Z = 4.31, P < .0001). Examination of the similarities and differences between the training regimens gives insight into the development of more effective and efficient interventions. The purpose of this "Current Concepts" review is to highlight the relative effectiveness of these interventions in reducing anterior cruciate ligament injury rates and to evaluate the common training components between the training studies. In addition, the level of rigor of these interventions, the costs and the difficulty of implementation, the compliance with these interventions, and the performance benefits are discussed. This review summarizes conclusions based on evidence from the common components of the various interventions to discuss their potential to reduce anterior cruciate ligament injury risk and assess their potential for combined use in more effective and efficient intervention protocols. PMID- 16382008 TI - Kinematics of the posterior cruciate ligament/posterolateral corner-injured knee after reconstruction by single- and double-bundle intra-articular grafts. AB - BACKGROUND: Single- and double-bundle reconstructions have been proposed for the knee after combined posterior cruciate ligament/posterolateral corner injuries. HYPOTHESIS: The double-bundle posterior cruciate ligament reconstruction is superior to the single-bundle posterior cruciate ligament reconstruction with regard to restoration of normal knee kinematics to the posterior cruciate ligament/posterolateral corner-sectioned knee. STUDY DESIGN: Controlled laboratory study. METHODS: Kinematics of 8 fresh-frozen, cadaveric human knees were determined in the following conditions: intact, sectioned posterior cruciate ligament/posterolateral corner, single anterolateral bundle posterior cruciate reconstruction, and double-bundle posterior cruciate reconstruction. RESULTS: The sectioned knee demonstrated a posterior shift of the tibial neutral position and the abnormal posterior, varus, and external rotation laxities used clinically to define a combined posterior cruciate ligament/posterolateral corner injury. Both reconstructions restored the posterior laxity to levels that were not statistically different from those seen in the intact knee, but the double-bundle reconstruction more closely mimicked the posterior laxity profile of the intact knee, having statistically lower posterior laxities than did the single-bundle reconstruction at 30 degrees, 60 degrees, and 90 degrees of flexion (P < .05, analysis of variance, HSD test). The resting position of the tibia after double bundle reconstruction trended to be anteriorly subluxated relative to its position for the intact knee at flexion angles of 30 degrees and greater (P <.05, paired t test). Neither technique corrected the abnormal varus or external rotation laxities. CONCLUSION: With either single- or double-bundle reconstructions, additional posterolateral reconstruction is recommended to correct the external rotation laxity. CLINICAL RELEVANCE: Knowledge of the kinematics of the combined posterior cruciate ligament/posterolateral corner injured knee is important in the proper diagnosis of the injury and in the selection of the appropriate surgical reconstruction. PMID- 16382009 TI - Maturation leads to gender differences in landing force and vertical jump performance: a longitudinal study. AB - BACKGROUND: Female athletes have increased risk of anterior cruciate ligament rupture after the onset of puberty. HYPOTHESES: Male athletes would demonstrate a longitudinal increase in vertical jump height compared with female athletes. There would be longitudinal gender differences in ground-reaction forces and loading rates. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: Sixteen female and 17 male adolescent athletes were evaluated for 2 consecutive years. Subjects were included if they were classified as pubertal during the first year of testing and postpubertal during the second year. As subjects performed a drop vertical jump, ground-reaction force, and vertical jump height were measured. Data analysis consisted of a mixed design analysis of variance with post hoc analysis (paired t tests). RESULTS: The male athletes demonstrated increased vertical jump height with maturation (P < .001); female athletes did not. Boys significantly reduced their landing ground-reaction force (P = .005), whereas girls did not. Takeoff force decreased in girls (P = .003) but not in boys. Both boys and girls had decreased loading rates with maturation (P < .001); however, girls had higher loading rates than did boys at both stages of maturation (P = .037). CONCLUSION: Male athletes demonstrated a neuromuscular spurt as evidenced by increased vertical jump height and increased ability to attenuate landing force. The absence of similar adaptations in female athletes may be related to the increased risk of anterior cruciate ligament injury. PMID- 16382010 TI - Does subchondral bone affect the fate of osteochondral allografts during storage? AB - BACKGROUND: Osteochondral allografts currently are hypothermically stored for a minimum of 14 days to a maximum of 28 days before surgical implantation, making storage conditions increasingly important. Previous studies have suggested that graft deterioration during storage may result from degradative factors and residual marrow elements in the subchondral bone. HYPOTHESIS: Allografts stored with large bone-to-cartilage ratios will be compromised after prolonged storage compared with grafts with minimal bone. STUDY DESIGN: Controlled laboratory study. METHODS: Osteochondral plugs were harvested from 16 fresh human femoral condyles and randomly assigned to 1 of 3 groups based on bone-to-cartilage ratios: 1:1, 5:1, or 10:1. These ratios were considered on the basis that the 1:1 ratio is the minimum bone necessary to press-fit an allograft and 10:1 is the present ratio used by tissue banks for allograft storage. After 14 and 28 days of storage at 4 degrees C, the specimens were assessed for viability and viable cell density using confocal microscopy, proteoglycan synthesis by (35)SO4 incorporation, and glycosaminoglycan content. RESULTS: All grafts underwent a significant decline in viable cell density, proteoglycan synthesis, and chondrocyte viability (particularly in the superficial region) after 14 days of storage, but no differences were observed between the 1:1, 5:1, or 10:1 ratio groups at either day 14 or day 28. In addition, no significant difference was noted in the glycosaminoglycan content in any of the groups. CONCLUSION: Osteochondral allografts stored with a 10:1 bone-to-cartilage ratio, similar to tissue-banking ratios, performed no worse than allografts stored with minimal bone, suggesting that the bone-to-cartilage ratio plays little to no role in the degradation of allografts during prolonged storage. CLINICAL RELEVANCE: As the practice of osteochondral allograft resurfacing becomes more commonplace, it is important that surgeons understand the factors that affect graft quality. PMID- 16382011 TI - Gender differences in outcome after anterior cruciate ligament reconstruction with hamstring tendon autograft. AB - BACKGROUND: It is now well documented that women are more likely to suffer anterior cruciate ligament injuries than are men. A few studies have examined gender differences in the outcome of anterior cruciate ligament reconstruction with patellar tendon graft and hamstring tendon with EndoButton fixation, but no well-controlled studies have specifically compared men and women after anterior cruciate ligament reconstruction with hamstring tendon graft and interference screw fixation. HYPOTHESIS: There is no difference in outcome between men and women after anterior cruciate ligament reconstruction with hamstring tendon autograft and interference screw fixation. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: There were 100 men and 100 women who underwent isolated anterior cruciate ligament reconstruction by a single surgeon. Patients were assessed preoperatively and at 1, 2, and 7 years after surgery. Variables were compared between female and male patients. RESULTS: Laxity on physical evaluation was greater in women than in men on Lachman (P = .04), pivot-shift (P = .05), and mean manual maximum testing (P = .05) at 7 years. However, the magnitude of this difference was small. No patient had a greater than grade 1 Lachman or pivot shift test result, and the mean difference between men and women was 0.6 mm on instrumented testing. Anterior cruciate ligament graft rupture occurred in 11 men and 10 women. There was no difference between male and female patients for self reported knee function or symptoms of instability or radiologic examination. CONCLUSION: Anterior cruciate ligament reconstruction using hamstring tendon autograft affords excellent self-reported and objective results in both men and women after 7 years. Although significantly greater laxity on physical examination was present in female patients than in male patients, the magnitude of this difference was small and had no effect on activity level, graft failure, or subjective or functional assessment. PMID- 16382012 TI - Range of joint movement in female dancers and nondancers aged 8 to 16 years: anatomical and clinical implications. AB - BACKGROUND: Little data are available on changes that occur with age in joint range of motion in dancers and nondancers. HYPOTHESIS: In dancers, joint range of motion will increase with age, whereas it will decrease in nondancers, independent of the joint studied. STUDY DESIGN: Cross-sectional study; Level of evidence, 3. METHODS: The study population included 1320 female dancers, aged 8 to 16 years, who participated in different types of dancing classes (classical ballet, modern dance, jazz, etc) and 226 nondancers of similar age. Range of motion was measured for the hip, knee, ankle, foot, and spinal joints. RESULTS: The pattern of differences in range of motion with age varied in different joints and types of movement. (1) For combined ankle and foot plantar flexion (pointe), ankle plantar flexion, and hip external rotation, there was no change in range of motion in dancers, whereas range of motion diminished with age in the nondancers. (2) For ankle dorsiflexion, neither group showed any change with age, and range of motion was significantly greater in the nondancer group. (3) For knee flexion, hip flexion, and hip internal rotation, range of motion decreased with age in both groups. (4) For hip abduction, range of motion decreased with age in dancers and remained constant in the nondancers. (5) For hip extension, range of motion increased in both groups. (6) For lower back and hamstrings, range of motion increased among dancers with age and remained constant among nondancers. CONCLUSION: Dancers and teachers should realize that passive joint range of motion is unlikely to improve with age. Therefore, the major goal of a dancing program should focus on exercises that retain the natural flexibility of the dancers' joints rather than trying to improve them. PMID- 16382013 TI - Shock attenuation of various protective devices for prevention of fall-related injuries of the forearm/hand complex. AB - BACKGROUND: Attenuation of the peak impact force is essential in any protective devices for prevention of fall-related injuries. HYPOTHESIS: Common wrist guards have limited effectiveness because of the multifaceted nature of wrist injury mechanisms, and other modalities may provide enhanced shock-absorbing functions. STUDY DESIGN: Controlled laboratory study. METHODS: A free-fall device was constructed using a mechanical surrogate to simulate falling impact. At 4 different falling heights, 5 different hand conditions were tested: bare hand, a generic-brand wrist guard, a Sorbothane glove, an air cell, and an air bladder condition. The impact force from the ground and the transmitted impact force to the forearm/hand complex were simultaneously measured. RESULTS: The falling height and hand condition significantly modulated the impact responses. The padded conditions always had significantly smaller peak impact forces compared with the bare-hand condition. The wrist guard became ineffective in impact force attenuation beyond the falling height of 51 cm. On the other hand, the air bladder condition maintained less than 45% of the peak impact force of the bare hand condition and remained below the critical value, whereas other conditions were all ineffective. CONCLUSION: It was reconfirmed that common wrist guard design could provide limited impact force attenuation, whereas damped pneumatic springs would provide substantially enhanced shock-absorbing functions. CLINICAL RELEVANCE: A wrist guard incorporating volar padding with the pneumatic spring design principle might be more effective at preventing injuries than are currently available designs. PMID- 16382014 TI - Measuring T cell alloreactivity to predict kidney transplant outcomes: are we there yet? PMID- 16382015 TI - Endothelial dysfunction contributes to renal function-associated cardiovascular mortality in a population with mild renal insufficiency: the Hoorn study. AB - Mildly impaired renal function is associated with cardiovascular morbidity and mortality. There are indications that endothelial dysfunction and/or chronic inflammation, which play an important role in atherothrombosis, are present in early stages of renal insufficiency. This study investigated whether and to which extent endothelial dysfunction and inflammation were related to renal function and contributed to renal function-associated cardiovascular mortality in a population-based cohort (n = 613), aged 50 to 75 yr, that was followed with a median duration of 12.5 yr. During follow-up, 192 individuals died (67 of cardiovascular causes). At baseline, renal function was estimated with serum creatinine, the Cockcroft-Gault formula, and the Modification of Diet in Renal Disease equation of GFR (eGFR). Endothelial function was estimated by plasma von Willebrand factor, soluble vascular cell adhesion molecule-1, and the urinary albumin-creatinine ratio. Inflammatory activity was estimated by plasma C reactive protein and soluble intercellular adhesion molecule-1. Renal function was mildly impaired (mean eGFR 68 +/- 12 ml/min per 1.73 m(2)) and independently associated with von Willebrand factor (standardized beta -0.09; 95% confidence interval [CI] -0.18 to -0.002; P < 0.05), soluble vascular cell adhesion molecule 1 (standardized beta -0.14; 95% CI -0.22 to -0.05; P < 0.01), and albumin creatinine ratio (standardized beta -0.15; 95% CI -0.23 to -0.08; P < 0.001) but not with markers of inflammatory activity. Renal function was inversely associated with cardiovascular and all-cause mortality. The relative risk for cardiovascular mortality but not all-cause mortality associated with renal function decreased from 1.22 to 1.12 per 5 ml/min per 1.73 m(2) decrease of eGFR after adjustment for markers of endothelial dysfunction. In conclusion, endothelial dysfunction was related to renal function and contributed to the excess in cardiovascular mortality in this population-based cohort with mild renal insufficiency. PMID- 16382016 TI - Cellular effects of guanylin and uroguanylin. AB - Ingestion of a salty meal induces secretion of guanylin (GN) and uroguanylin (UGN) into the intestinal lumen, where they inhibit Na+ absorption and induce Cl , HCO3-, and water secretion. Simultaneously, these hormones stimulate renal electrolyte excretion by inducing natriuresis, kaliuresis, and diuresis. GN and UGN therefore participate in the prevention of hypernatremia and hypervolemia after salty meals. The signaling pathway of GN and UGN in the intestine is well known. They activate enterocytes via guanylate cyclase C (GC-C), which leads to cGMP-dependent inhibition of Na+/H+ exchange and activation of the cystic fibrosis transmembrane regulator. In GC-C-deficient mice, GN and UGN still produce renal natriuresis, kaliuresis, and diuresis, suggesting different signaling pathways in the kidney compared with the intestine. Signaling pathways for GN and UGN in the kidney differ along the various nephron segments. In proximal tubule cells, a cGMP- and GC-C-dependent signaling was demonstrated for both peptides. In addition, UGN activates a pertussis toxin-sensitive G-protein coupled receptor. A similar dual signaling pathway is also known for atrial natriuretic peptide. Recently, a cGMP-independent signaling pathway for GN and UGN was also shown in principal cells of the human and mouse cortical collecting duct. Because GN and UGN activate different signaling pathways in specific organs and even within the kidney, this review focuses on more recent findings on cellular effects and signaling mechanisms of these peptides and their pathophysiologic implications in the intestine and the kidney. PMID- 16382017 TI - Apolipoprotein A-IV predicts progression of chronic kidney disease: the mild to moderate kidney disease study. AB - It has not been established firmly whether dyslipidemia contributes independently to the progression of kidney disease. Lipid and lipoprotein parameters, including levels of total, HDL, and LDL cholesterol; triglycerides; lipoprotein(a); apolipoprotein A-IV; and the apolipoprotein E and A-IV polymorphisms, were assessed in 177 patients who had mostly mild to moderate renal insufficiency and were followed prospectively for up to 7 yr. Progression of kidney disease was defined as doubling of baseline serum creatinine and/or terminal renal failure necessitating renal replacement therapy. In univariate analysis, patients who reached a progression end point (n = 65) were significantly older and had higher serum creatinine and proteinuria as well as lower GFR and hemoglobin levels. In addition, baseline apolipoprotein A-IV and triglyceride concentrations were higher and HDL cholesterol levels were lower. Multivariate Cox regression analysis revealed that baseline GFR (hazard ratio 0.714; 95% confidence interval [CI] 0.627 to 0.814 for an increment of 10 ml/min per 1.73 m(2); P < 0.0001) and serum apolipoprotein A-IV concentrations (hazard ratio 1.062; 95% CI 1.018 to 1.108 for an increment of 1 mg/dl; P = 0.006) were significant predictors of disease progression. Patients with apolipoprotein A-IV levels above the median had a significantly faster progression (P < 0.0001), and their mean follow-up time to a progression end point was 53.7 mo (95% CI 47.6 to 59.8) as compared with 70.0 mo (95% CI 64.6 to 75.4) in patients with apolipoprotein A-IV levels below the median. For the apolipoprotein E polymorphism, only the genotype epsilon2/epsilon4 was associated with an increased risk for progression. In summary, this prospective study in patients with nondiabetic primary kidney disease demonstrated that apolipoprotein A-IV concentration is a novel independent predictor of progression. PMID- 16382018 TI - Cross-sectional association of kidney function with valvular and annular calcification: the Framingham heart study. AB - Valvular calcification is common in the setting of end-stage kidney disease and is associated with increased risks for cardiovascular disease events. It is unknown whether the prevalence of valvular calcification is increased in milder kidney disease after accounting for cardiovascular risk factors. Participants who attended the sixth examination of the Framingham Offspring Study (1995 to 1998) were eligible. Kidney function was estimated by GFR using the simplified Modification of Diet in Renal Disease Study equation. Mitral annular calcification (MAC), aortic sclerosis, and aortic annular calcification were assessed by two-dimensional echocardiography. Logistic regression was used to examine the odds of valvular calcification among participants with chronic kidney disease (CKD; GFR < 60 ml/min per 1.73 m(2)). A total of 3047 participants (52% women; mean age 59 +/- 10 yr) were available for analysis. CKD was present in 8.6% (n = 262) of the sample. Among participants with valve/annular calcification (n = 284; 9.3%), 20% had CKD, compared with 7% in patients without valvular calcification. After adjustment for age, gender, systolic and diastolic BP, hypertension treatment, total/HDL cholesterol, body mass index, diabetes, smoking status, and cardiovascular disease, participants with CKD had a 60% increased odds of MAC (odds ratio 1.6; 95% confidence interval 1.03 to 2.5). There was no significant association between CKD and either aortic sclerosis or aortic annular calcification (odds ratio 1.1 and 1.1, respectively). After age and gender adjustment, the combination of both CKD and MAC was associated with a three-fold increased risk for death compared with those with neither condition (P = 0.0004). In the community, CKD is associated with presence of MAC before the onset of ESRD. Further research is warranted to understand whether traditional and novel vascular risk factor burden, as well as metabolic derangements found in early kidney disease, can account for the CKD-MAC association. PMID- 16382019 TI - Compartment-specific expression and function of the chemokine IP-10/CXCL10 in a model of renal endothelial microvascular injury. AB - The recruitment of inflammatory cells into renal tissue, mainly T cells and monocytes, is a typical feature of various renal diseases such as glomerulonephritis, thrombotic angiopathies, allograft rejection, and vasculitis. T cells predominantly infiltrate the tubulointerstitium, whereas monocytes are present in the tubulointerstitial and glomerular compartment. Because chemokines play a pivotal role in leukocyte trafficking under inflammatory conditions, this study investigated whether a differential expression of chemokines contributes to the precise coordination of leukocyte subtype trafficking in a rat model of renal microvascular endothelial injury. Renal microvascular endothelial injury was induced in rats by selective renal artery perfusion with an anti-endothelial antibody. Induction of the disease led to severe glomerular and tubulointerstitial endothelial injury with subsequent upregulation of chemokines followed by inflammatory cell recruitment. Among the analyzed chemokine mRNA, IP 10/CXCL10 (119-fold), acting via CXCR3 on activated T cells, and MCP-1/CCL2 (65 fold), acting via CCR2 on monocytes, were by far the most strongly upregulated chemokines. In situ hybridization revealed that IP-10/CXCL10 mRNA was selectively expressed by endothelial cells in the tubulointerstitial area, co-localizing with infiltrating T cells. Despite extensive damage of glomerular vasculature, no IP 10/CXCL10 expression by glomerular endothelial cells was detected. MCP-1/CCL2 mRNA in contrast was detectable in the glomerulus and the tubulointerstitium. Treatment with a neutralizing anti-IP-10/CXCL10 antibody significantly reduced the number of infiltrating tubulointerstitial T cells without affecting monocyte migration and led to an improved renal function. Our study demonstrates a role of IP-10/CXCL10 on T cell recruitment in a rat model of renal endothelial microvascular injury. Furthermore, a differential chemokine expression profile by endothelial cells in different renal compartments was found. These findings are consistent with the hypothesis that functional heterogeneity of endothelial cells from different vascular sites exists and provide an insight into the molecular mechanisms that may mediate compartment-specific T cell and monocyte recruitment in inflammatory renal disease. PMID- 16382020 TI - Panel of reactive T cells as a measurement of primed cellular alloimmunity in kidney transplant candidates. AB - Pretransplantation panel reactive antibody (PRA) testing assesses posttransplantation risk for antibody-mediated graft injury. It was postulated analogously that screening for effector/memory alloreactive T cells by "panel of reactive T cells" (PRT) using IFN-gamma enzyme-linked immunosorbent spot assays would evaluate independently cellular alloimmunity in transplant candidates. Peripheral blood lymphocytes from 41 hemodialysis patients who were awaiting first renal transplants were tested against a panel of allogeneic stimulator cells. Positive assays were defined arbitrarily as >25 spots/300,000 peripheral blood lymphocytes, and positive PRT was defined as when the responder reacted to 40 or 75% (PRT-75) of the stimulators. Seventeen percent of patients were PRT 75+, whereas 32% were PRA+. Twelve percent of the cohort was PRT-75+/PRA-, and only 5% of the patients were PRA+/PRT-75+, indicating that T cell alloreactivity did not routinely imply B cell sensitization and vice versa. PRT-75+ patients were more likely to be younger (<55 yr) and black. In contrast, a positive PRA was significantly associated with female gender but not race or age. Pretransplantation screening of cellular alloimmunity by enzyme-linked immunosorbent spot-based PRT detects a subset of hemodialysis patients who differ from those that are PRA+. Preliminary correlations with posttransplantation outcome in seven recipients suggest that PRT screening has the potential to aid in risk assessment in renal transplant candidates. PMID- 16382021 TI - Serum beta-2 microglobulin levels predict mortality in dialysis patients: results of the HEMO study. AB - In the randomized Hemodialysis (HEMO) Study, chronic high-flux dialysis, as defined by higher beta-2 microglobulin (beta(2)M) clearance, compared with low flux dialysis did not significantly alter all-cause mortality in the entire cohort but was associated with lower mortality in long-term dialysis patients. This analysis examined the determinants of serum beta(2)M levels and the associations of serum beta(2)M levels or dialyzer beta(2)M clearance with mortality. In a multivariable regression model that examined 1704 patients, baseline residual kidney urea clearance and dialyzer beta(2)M clearance were strong predictors of predialysis serum beta(2)M levels at 1 mo of follow-up, with regression coefficients of -7.21 (+/-0.69 SE) mg/L per ml/min per 35 L urea volume (P < 0.0001) and -1.94 (+/-0.30) mg/L per ml/min (P < 0.0001),respectively. In addition, black race and baseline years on dialysis correlated positively whereas age, diabetes, serum albumin, and body mass index correlated negatively with serum beta(2)M levels (P < 0.05). In time-dependent Cox regression models, mean cumulative predialysis serum beta(2)M levels but not dialyzer beta(2)M clearance were associated with all-cause mortality (relative risk = 1.11 per 10-mg/L increase in beta(2)M level; 95% confidence interval 1.05 to 1.19; P = 0.001), after adjustment for residual kidney urea clearance and number of prestudy years on dialysis. This association is supportive of the potential value of beta(2)M as a marker to guide chronic hemodialysis therapy. PMID- 16382022 TI - IFN-inducible protein-10 plays a pivotal role in maintaining slit-diaphragm function by regulating podocyte cell-cycle balance. AB - IFN-inducible protein-10 (IP-10/CXCL10) is a potent chemoattractant for activated T lymphocytes and was reported recently to have several additional biologic activities. In this study, the pathophysiologic role of IP-10 in the glomerular visceral epithelial cell (podocyte) was investigated. In cultured podocytes subjected to recombinant IP-10 treatment, the expression of slit-diaphragm (SD) components nephrin and podocin clearly was heightened. Rats that had puromycin aminonucleoside nephropathy and anti-nephrin antibody-induced nephropathy and were subjected to anti-IP-10 function-blocking antibody (anti-IP-10 mAb) treatment displayed a decrease in the protein level of SD components, as well as exacerbated proteinuria. For exploration of the mechanisms of this process, the interaction between IP-10 and the cell-cycle regulatory proteins was investigated. Cultured podocytes subjected to recombinant IP-10 treatment displayed an increase in the protein level of p27(Kip1), whereas the levels of cyclins E and A decreased. The expression of IP-10 and SD components was heightened by the treatment of siRNA of cyclin A, whereas these expressions were lowered by the treatment of siRNA of p27(Kip1). Proteinuric rats subjected to anti-IP-10 mAb treatment displayed a heightened expression of cyclin A from the early phase of the disease, which indicates that the anti-IP-10 mAb treatment exacerbates podocyte injury by disturbing the cell-cycle balance. These results raise the possibility that IP-10 could become a novel therapeutic target in nephrotic syndrome and several diseases with altered cell-cycle balance. PMID- 16382023 TI - Reciprocal interaction of serotonin and neuronal activity in regulation of cAMP responsive element-dependent gene expression. AB - Neuronal activity triggers multiple signal transduction pathways and potently regulates gene expression in the brain. In the central nervous system, in addition to the synaptic input, neurons are subject to neuromodulatory influences that can activate the same signaling elements. However, the principles that govern the interaction of neuromodulators and neuronal activity in the regulation of gene expression are unclear. Here, we examine how serotonergic neuromodulation interacts with neuronal activity in the regulation of gene expression in hippocampal neurons. We show that cAMP-response element binding protein (CREB) phosphorylation and gene expression were stimulated by serotonin (5-HT) in the absence of neuronal activity. In contrast, in the presence of neuronal activity, 5-HT inhibited gene expression down to the baseline, although neuronal activity alone was sufficient to maximally activate gene expression. The ability of 5-HT to stimulate CREB phosphorylation in the absence of neuronal activity or inhibit CREB phosphorylation during activity was due to a tight balance between protein kinases and phosphatases that could be physiologically tilted by different serotonergic receptors or exogenously influenced by phosphatase and kinase inhibitors. Taken together, these results suggest a reciprocal inhibitory interaction between neuronal activity and 5-HT in the regulation of cAMP response element-dependent gene expression in hippocampal neurons. PMID- 16382024 TI - Inhibition of neprilysin by infusion of thiorphan into the hippocampus causes an accumulation of amyloid Beta and impairment of learning and memory. AB - An imbalance between anabolism and catabolism causes an accumulation of amyloid beta-peptide (Abeta), which is a proposed trigger of the onset of Alzheimer's disease. Neprilysin is a rate-limiting peptidase that participates in the catabolism of Abeta in the brain. We examined whether rats continuously infused with thiorphan, a specific neprilysin inhibitor, into the hippocampus develop cognitive impairments through accumulation of Abeta. Thiorphan infusion elevated hippocampal Abeta40 and Abeta42 levels in the insoluble but not the soluble fraction. Thiorphan-infused rats displayed cognitive impairments in the ability to discriminate in the object recognition test, associative learning in the conditioned fear learning test, and spatial memory in the water maze test, tasks that depend on the hippocampus. These cognitive abilities in the battery of behavioral tasks inversely correlated with insoluble Abeta contents in the hippocampus. The nicotine-stimulated release of acetylcholine in the hippocampus of thiorphan-infused rats was significantly lower than that in vehicle-infused rats. These results indicate that continuous infusion of thiorphan into the hippocampus causes cognitive dysfunction and reduces cholinergic activity by raising the level of Abeta in the hippocampus and suggest that a reduction of neprilysin activity contributes to the deposition of Abeta and development of Alzheimer's disease. PMID- 16382025 TI - The phase 2 enzyme inducers ethacrynic acid, DL-sulforaphane, and oltipraz inhibit lipopolysaccharide-induced high-mobility group box 1 secretion by RAW 264.7 cells. AB - The diuretic ethacrynic acid (EA) has been shown to inhibit signaling by the proinflammatory transcription factor nuclear factor-kappaB (NF-kappaB). Accordingly, we sought to determine whether this compound is capable of inhibiting the release of cytokines [interleukin (IL)-6 and IL-10] and NO from RAW 264.7 murine macrophage-like cells stimulated with lipopolysaccharide (LPS). Additionally, we sought to determine whether EA can inhibit secretion of high mobility group box 1 (HMGB1), a nuclear protein that is secreted by immunostimulated macrophages and functions in the extracellular milieu as a proinflammatory mediator. In a concentration-dependent manner, EA inhibited secretion of IL-6, IL-10, nitric oxide, and HMGB1. As expected, EA inhibited NF kappaB DNA binding in LPS-stimulated RAW 264.7 cells. Treating these cells with pyrrolidine dithiocarbamate, SN50 (amino acid sequence AAVALLPAVLLALLAPVQRKRQKLMP) or 5-(thien-3-yl)-3-aminothiophene-2-carboxamide (SC 514) also inhibited LPS-induced NF-kappaB DNA binding, but these compounds failed to inhibit LPS-induced HMGB1 secretion. These findings suggested that inhibition of HMGB1 secretion by EA might occur via a mechanism unrelated to the NF-kappaB signaling pathway. Because EA is an electrophilic compound that is known to be capable of inducing expression of so-called phase 2 proteins, we sought to determine whether two other phase 2 enzyme inducers, oltipraz and DL sulforaphane, also are capable of inhibiting HMGB1 release from immunostimulated macrophages. Incubating RAW 264.7 cells with either oltipraz or DL-sulforaphane inhibited LPS-induced HMGB1 secretion. Moreover, both EA and DL-sulforaphane inhibited relocalization of nuclear HMGB1 into the cytoplasm of LPS-stimulated RAW 264.7 cells. These data suggest that phase 2 inducers may exert anti inflammatory effects by inhibiting secretion of the cytokine-like nuclear protein HMGB1. PMID- 16382026 TI - Deficiency of SPAG16L causes male infertility associated with impaired sperm motility. AB - The axonemes of cilia and flagella contain a "9+2" structure of microtubules and associated proteins. Proteins associated with the central doublet pair have been identified in Chlamydomonas that result in motility defects when mutated. The murine orthologue of the Chlamydomonas PF20 gene, sperm-associated antigen 16 (Spag16), encodes two proteins of M(r) approximately 71 x 10(3) (SPAG16L) and M(r) approximately 35 x 10(3) (SPAG16S). In sperm, SPAG16L is found in the central apparatus of the axoneme. To determine the function of SPAG16L, gene targeting was used to generate mice lacking this protein but still expressing SPAG16S. Mutant animals were viable and showed no evidence of hydrocephalus, lateralization defects, sinusitis, bronchial infection, or cystic kidneys symptoms typically associated with ciliary defects. However, males were infertile with a lower than normal sperm count. The sperm had marked motility defects, even though ultrastructural abnormalities of the axoneme were not evident. In addition, the testes of some nullizygous animals showed a spermatogenetic defect, which consisted of degenerated germ cells in the seminiferous tubules. We conclude that SPAG16L is essential for sperm flagellar function. The sperm defect is consistent with the motility phenotype of the Pf20 mutants of Chlamydomonas, but morphologically different in that the mutant algal axoneme lacks the central apparatus. PMID- 16382027 TI - The first mitosis of the mouse embryo is prolonged by transitional metaphase arrest. AB - The first mitosis of the mouse embryo is almost twice as long as the second. The mechanism of the prolongation of the first mitosis remains unknown, and it is not clear whether prometaphase or metaphase or both are prolonged. Prometaphase is characterized by dynamic chromosome movements and spindle assembly checkpoint activity, which prevents anaphase until establishment of stable kinetochore microtubule connections. The end of prometaphase is correlated with checkpoint inactivation and disappearance of MAD2L1 (MAD2) and RSN (CLIP-170) proteins from kinetochores. Spindle assembly checkpoint operates during the early mouse mitoses, but it is not clear whether it influences their duration. Here, we determine the length of prometaphases and metaphases during the first two embryonic mitoses by time-lapse video recording of chromosomes and by immunolocalization of MAD2L1 and RSN proteins. We show that the duration of the two prometaphases does not differ and that MAD2L1 and RSN disappear from kinetochores very early during each mitosis. The first metaphase is significantly longer than the second one. Therefore, the prolongation of the first embryonic mitosis is due to a prolonged metaphase, and the spindle assembly checkpoint cannot be involved in this process. We show also that MAD2L1 staining disappears gradually from kinetochores of oocytes arrested at metaphase of the second meiotic division. This shows a striking similarity between the first embryonic mitosis and metaphase arrest in oocytes. We postulate that the first embryonic mitosis is prolonged by a transient metaphase arrest that is independent of the spindle assembly checkpoint and is similar to metaphase II arrest. The molecular mechanism of this transient arrest remains to be elucidated. PMID- 16382028 TI - Violence against American Indian Women and the Services-Training-Officers Prosecutors Violence Against Indian Women (STOP VAIW) program. AB - When Congress appropriated funds to develop ways to reduce violence against American Indian women, tribal elders faced a challenging task: find ways to cooperate with various tribal and nontribal criminal justice agencies and navigate the maze of law enforcement authority. An evaluative study was conducted of these programs and the different approaches used to help keep women safe by American Indian tribal governments. This study found that the tribes rose to the challenge; the money was making a difference. The grants to stop violence against Indian women have made a significant impact in the 134 native communities that received awards. PMID- 16382029 TI - Evaluation of a police and social services domestic violence program: empirical evidence needed to inform public health policies. AB - The Family Violence Response Team (FVRT) responded to police calls for domestic violence and provided services to victims. Police records were followed for (a) 327 FVRT clients with an index police visit in 1998 and (b) 498 nonconcurrent controls with an index visit in 1997. Except for marriage, no demographic characteristics were associated with batterer recidivism, as measured by police calls. The between-group odds ratio (OR) suggested that FVRT clients experienced a 1.7 (95% Confidence Interval [CI]: 1.2 to 2.5) times greater recidivism rate than controls. Although increased reporting cannot be ruled out, results raise concerns about the effects of domestic violence interventions. PMID- 16382030 TI - Protecting prosecution: exploring the powers of law in an intervention program for domestic violence. AB - This article critically analyzes how the criminal justice system centrally situates itself in an intervention program intended to protect victims of domestic abuse and stalking. Based on the first empirical, in-depth study in the Netherlands of an intervention program using electronic technology that is increasingly used in the United States, results indicate how the central role of the criminal justice actors can evoke a shift toward foregrounding prosecutorial interests. Drawing from a critical theoretical understanding of the powers of law and the legal system, the author argues that current tendencies toward criminalization in domestic violence interventions can have an unintended violent impact for victims who are either excluded from the program or are forced into a criminal justice regime that might not be in their primary interest. In this study, women seem to actively navigate the use of the criminal justice system to receive the protection they need. PMID- 16382031 TI - Interpersonal violence at the crossroads between adolescence and adulthood: learning about partner violence from young mothers. AB - This article examines partner violence at the crossroads between adolescence and adulthood by talking with adolescent mothers--young women who straddle that crossroad. Drawing on ethnographic interviews conducted with 30 adolescent mothers, the accounts of 13 young mothers were analyzed to shed light on the ways in which their experiences of partner violence intersected with their developmental stage. Thematic analysis produced four general categories of themes that included physical proximity of family members, chaos at home, growing up in the shadow of abuse, and making the relationship work. Implications for future research and practice are explored. PMID- 16382032 TI - CSF findings in 250 patients with serologically confirmed West Nile virus meningitis and encephalitis. AB - OBJECTIVE: To provide a large, comprehensive evaluation of the CSF findings in patients with serologically confirmed West Nile virus (WNV), CNS disease, and their correlation with outcome. METHODS: CSF samples from 334 WNV-infected hospitalized patients were analyzed. Information was available and extracted from the medical records of 250 of these patients, and CSF parameters correlated with clinical and epidemiologic features of disease (e.g., patient age, sex, outcome). RESULTS: Patients with meningitis had a mean of 226 cells/mm3, and those with encephalitis had a mean of 227 cells/mm3. Three percent of meningitis patients and 5% of encephalitis patients had fewer than 5 cells/mm3, and approximately 8% of both groups had more than 500 cells/mm3. Patients with meningitis had a mean of 41% neutrophils, and those with encephalitis had 45%. Forty-five percent of meningitis patients and 37% of encephalitis patients had at least 50% neutrophils in their initial CSF specimen. Neither the mean percent neutrophils nor their distribution differed significantly between groups. Forty-seven percent of encephalitis patients and 16% of meningitis patients had CSF protein of 100 mg/dL or greater (p < 0.01). Although specific CSF parameters, including nucleated cell count and protein concentration, correlated significantly with outcome, multivariate analysis suggested that their total predictive value was modest. Age was an additional predictor of outcome independent of CSF variables in all patients. CONCLUSIONS: Serologically confirmed West Nile virus meningitis and encephalitis produce similar degrees of CSF pleocytosis and are frequently associated with substantial CSF neutrophilia. Patients with encephalitis have higher CSF protein concentrations and are more likely to have adverse outcomes, including admission to long-term care facilities or even death after their acute illness. CSF findings were only a modest predictor of disease outcome, with patient age adding important independent prognostic information. PMID- 16382033 TI - Premorbid antiplatelet use and ischemic stroke outcomes. AB - OBJECTIVE: To evaluate the independent effect of premorbid antiplatelet use on incident ischemic stroke severity and outcome at discharge. METHODS: The authors studied consecutive patients presenting within 24 hours of ischemic stroke over a 1-year period. National Institutes of Health Stroke Scale (NIHSS) score at presentation was used as index of stroke severity and a modified Rankin scale of 0 to 1 at discharge as index of good functional outcome. Patients were categorized according to their premorbid antiplatelet use as antiplatelet inclusive (AI) and no antiplatelet (NA). Demographic data, risk factors, pertinent laboratory tests, other medications, and stroke mechanisms were controlled for across the two groups using multivariate logistic regression. RESULTS: A total of 260 individuals met study criteria: 92 patients were on antiplatelet agents prior to admission, 168 were on no antiplatelets. Pretreatment with antiplatelet was associated with lower presenting median NIHSS (4.5 vs 7, p = 0.005). Antiplatelet use was associated with less severe stroke at presentation in those having no history of stroke or TIA (4.8 vs 8.0, p = 0.03) but not in those with a prior history of stroke or TIA (4.9 vs 4.9, p = 0.987). The likelihood of a good outcome was increased in those on antiplatelets after adjusting for other variables (OR 2.105, p = 0.0073). CONCLUSIONS: Prestroke use of antiplatelet may be associated with reduced severity of incident ischemic strokes in those with no prior history of stroke or TIA, and with an increased likelihood of a good discharge outcome regardless of prior cerebrovascular event history. PMID- 16382034 TI - Seizure control and treatment in pregnancy: observations from the EURAP epilepsy pregnancy registry. AB - OBJECTIVE: To analyze seizure control and treatment in pregnant women with epilepsy. METHODS: Seizure control and treatment were recorded prospectively in 1,956 pregnancies of 1,882 women with epilepsy participating in EURAP, an international antiepileptic drugs (AEDs) and pregnancy registry. RESULTS: Of all cases, 58.3% were seizure-free throughout pregnancy. Occurrence of any seizures was associated with localization-related epilepsy (OR: 2.5; 1.7 to 3.9) and polytherapy (OR: 9.0; 5.6 to 14.8) and for tonic-clonic seizures, with oxcarbazepine monotherapy (OR: 5.4; 1.6 to 17.1). Using first trimester as reference, seizure control remained unchanged throughout pregnancy in 63.6%, 92.7% of whom were seizure-free during the entire pregnancy. For those with a change in seizure frequency, 17.3% had an increase and 15.9% a decrease. Seizures occurred during delivery in 60 pregnancies (3.5%), more commonly in women with seizures during pregnancy (OR: 4.8; 2.3 to 10.0). There were 36 cases of status epilepticus (12 convulsive), which resulted in stillbirth in one case but no cases of miscarriage or maternal mortality. AED treatment remained unchanged in 62.7% of the pregnancies. The number or dosage of AEDs were more often increased in pregnancies with seizures (OR: 3.6; 2.8 to 4.7) and with monotherapy with lamotrigine (OR: 3.8; 2.1 to 6.9) or oxcarbazepine (OR: 3.7; 1.1 to 12.9). CONCLUSIONS: The majority of patients with epilepsy maintain seizure control during pregnancy. The apparently higher risk of seizures among women treated with oxcarbazepine and the more frequent increases in drug load in the oxcarbazepine and lamotrigine cohorts prompts further studies on relationships with pharmacokinetic changes. Risks associated with status epilepticus appear to be lower than previously reported. PMID- 16382035 TI - Albendazole trial at 15 or 30 mg/kg/day for subarachnoid and intraventricular cysticercosis. AB - Thirty-six patients with subarachnoid and intraventricular cysticercosis were randomly assigned to receive albendazole at 15 or 30 mg/kg/day plus dexamethasone for 8 days. Results favored a higher dose, with larger cyst reduction on MRI at 90 and 180 days and higher albendazole sulfoxide levels in plasma. An albendazole course at 30 mg/kg/day combined with corticosteroids is safe and more effective than the usual dose. A single treatment was insufficient in intraventricular and giant cysts. PMID- 16382037 TI - Proteomics in tumor progression and metastasis. AB - With the ultimate goal of systematically identifying and characterizing proteins within an organism, the field of proteomics has generated much excitement in the past few years. Coupled with mass spectrometry, various quantitative and functional techniques are now available that allow for large-scale analyses of proteins implicated in cancer. New techniques are just now being applied to identifying the temporal changes in protein levels associated with tumor development. This review will focus on the use and promise of proteomic technologies as they apply to the study of tumor progression and metastasis. PMID- 16382038 TI - Cytoplasmic/nuclear shuttling and tumor progression. AB - Protein localization is a highly dynamic biological process. To ensure a proper cellular function, the spatial distribution of different proteins needs to be delicately regulated and coordinated. In cancer, this process is tightly correlated with the outcome of cell proliferation and the response to apoptotic stress. Here we summarize our recent studies regarding the role of subcellular trafficking on cancer cell growth through an AKT-independent regulation of the forkhead transcription factor and, on cancer metastasis, through the regulated localization and subsequent protein degradation of the Snail protein, a key transcription factor involved in epithelial-mesenchymal transition (EMT). In both cases, cellular signaling pathways mediated by distinct protein kinases modulate the appropriate protein localization and functioning. Finally, we address a novel translocation pathway for tyrosine kinase receptors between the cell membrane and the nucleus. We demonstrate that the ErbB family proteins are also expressed in the nucleus and can function as transcriptional regulators. Using a genomic approach, we have identified a number of the target genes regulated by this pathway that are closely related to cancer cell proliferation and metastasis. Our findings, together with the seminal discoveries by other groups, highlight the importance of protein localization in cancer biology and its potential to be a therapeutic target for cancer. PMID- 16382039 TI - Targeting intracellular signaling pathways as a novel strategy in melanoma therapeutics. AB - Melanoma has been one of the fastest rising malignancies in the last four decades with cases increasing from below 3 per 100,000 people to above 13. Despite worldwide efforts in prevention, diagnosis, and treatment, cases of melanoma continue to rise at an alarming rate of 2.5% annually in the United States. Although early primary melanomas are curable through surgery, treatment of advanced disease remains difficult and the strategies employed in the last 30 years have not significantly improved cure rates, which are less than 5%. The recent identification of activating mutations in BRAF in over 60% of cases of melanoma has caused much excitement in the melanoma community and may offer the first opportunity for a rational treatment program. Combination therapy using the RAF inhibitor, BAY 43-9006, and chemotherapy has led to impressive responses in some melanoma patients and provides a new paradigm for therapeutic intervention in this intractable disease. Besides activating mutations in BRAF, melanomas have constitutive activity in a number of other signaling pathways implicated in oncogenesis, including PI3 kinase/Akt, NFB, Src, and STAT3. With more and more selective small molecule inhibitors becoming available, there are good prospects for treating advanced melanoma using new combinations of signal transduction inhibitors and chemotherapy. In the current review, we discuss the role for these signaling pathways in melanoma and discuss the rationale for targeting signaling cascades using small molecule inhibitors. PMID- 16382040 TI - Studies into the anticancer effects of selenomethionine against human colon cancer. AB - Colorectal cancer is the third most frequent fatal malignant neoplasm in the United States and is expected to cause significant morbidity and mortality. The recent recall of cyclooxygenase-2 inhibitors from clinical trials highlights the need to develop other agents for cancer chemoprevention trials. Intervention strategies with selenium compounds represent a viable option to reduce colon cancer. Here we discuss epidemiologic studies and ongoing clinical trials with selenium. In addition, we discuss preclinical mechanistic studies that provide insights into the biochemical and molecular bases for the anticancer effects of selenomethionine. PMID- 16382041 TI - Mechanisms of prostate tumorigenesis: roles for transcription factors Nkx3.1 and Egr1. AB - Recent developments in the generation and analysis of transgenic mouse models have improved our understanding of the early stages of prostate tumorigenesis. Analysis of models based on the homeodomain protein Nkx3.1 and the zinc finger protein Egr1 suggests that these transcription factors play distinct roles in the initiation and progression of precursor prostatic intraepithelial neoplasia (PIN) lesions, respectively. Nkx3.1 is a candidate prostate tumor suppressor gene (TSG) that demonstrates haploinsufficiency. Disruption of one or both copies of the murine Nkx3.1 gene leads to the development of epithelial hyperplasia and PIN. This appears to be a consequence of delayed exit from the cell cycle by differentiating prostate luminal epithelial cells in Nkx3.1 mutant mice. Gene expression profiling has provided additional insight into the basis of haploinsufficiency in Nkx3.1 mutant mice. A reduction in Nkx3.1 dosage leads to dramatic alterations in the expression of a subset of genes by altering the probability of a target gene existing in the "on" or "off" state. The immediate early gene Egr1, on the other hand, is overexpressed in human and mouse prostate tumors and PIN lesions and regulates the expression of several genes implicated in prostate tumor progression, including platelet-derived growth factor and insulin-like growth factor II. Prostate cancer-prone mice lacking Egr1 exhibit a significant delay in tumor progression. Specifically, Egr1 deficiency impairs the transition from PIN to invasive carcinoma. Thus, Nkx3.1 and Egr1 regulate gene programs involved in distinct aspects of prostate tumorigenesis. PMID- 16382042 TI - Sulindac sulfone is most effective in modulating beta-catenin-mediated transcription in cells with mutant APC. AB - Sulindac sulfone (FGN-1, Aptosyn), a metabolite of the nonsteroidal anti inflammatory drug sulindac, lacks cyclooxygenase inhibitory activity. Although its ability to inhibit tumorigenesis in both carcinogen-treated animals and patients with familial adenomatous polyposis has been attributed to the induction of apoptosis, its complete mechanism of action remains unclear. The purpose of the present study was to determine the ability of sulindac metabolites to regulate cellular levels of beta-catenin and downstream targets of the adenomatous polyposis coli (APC)/beta-catenin pathway in vitro. Sulindac sulfone was consistently more potent than the sulfide metabolite in all analyses, significantly decreasing the expression of total cellular beta-catenin (50% of control), pro-caspase 3 (49%), cyclin D1 (51%), and PPARdelta (65%) in SW480 cells. No significant alteration in pro-caspase 3 or beta-catenin expression was found in HCA7, LS174, or Caco-2 cells treated with sulindac sulfone. A dose dependent reduction in TCF-mediated transcriptional activity was also observed in SW480 cells. These data demonstrate that sulindac sulfone can modulate the APC/beta-catenin pathway in vitro and that its efficacy is dependent upon the mutational status of APC and beta-catenin. PMID- 16382043 TI - Targeting ligand cleavage to inhibit the ErbB pathway in cancer. AB - ADAMs (a disintegrin and metalloproteases) are zinc-dependent transmembrane metalloproteases that shed the extracellular domains of membrane-bound growth factors, cytokines, and receptors. Recently, ADAMs have emerged in ErbB signaling pathways as sheddases or multiple ErbB ligands. As the ErbB pathway is a validated target for anticancer drugs, upstream activators of ErbB ligands, their sheddases, become new drug targets in the ErbB pathway. We have identified selective small molecule inhibitors of ADAM proteases that block shedding and activation of multiple ErbB ligands, and we are planning to test the compounds in the clinic. PMID- 16382044 TI - Preclinical modeling of combination treatments: fantasy or requirement? AB - Combination chemotherapy is considered the "norm" in cancer chemotherapy. While the development of classical combination regimens took into account factors such as single-agent activity, different toxicity profiles, and advantageous pharmacology of the component drugs, in the era of targeted therapies there are additional considerations such as new mechanisms of resistance due to modulation of pathway dependence. Since it is not feasible to test all of the possible combinations clinically, some method for preclinically identifying and prioritizing promising combinations is necessary. While in vivo animal models can be used for safety testing and some pharmacokinetics, even they can quickly become prohibitively resource intensive for the purpose of efficacy determinations. Therefore, factors such as biologic rationale, reliable in vitro results in more than one tumor type, achievable in vivo pharmacology, and selectivity of primary tumor cell activity become important in the evaluation of potential combination regimens. PMID- 16382045 TI - Mechanisms of trastuzumab resistance and their clinical implications. AB - Trastuzumab (Herceptin) is an excellent model of rationally designed targeted cancer treatment. However, less than 35% of patients with ErbB2-positive breast tumors respond to trastuzumab as a single agent, and 2-5% of trastuzumab-treated patients suffer from severe side effects, including cardiac dysfunction. Recent progress in understanding the mechanisms of trastuzumab antitumor function and cellular defects leading to trastuzumab resistance is summarized. Also explored is the potential of combination therapies for reversing trastuzumab resistance. PMID- 16382046 TI - Regulation of cancer cell survival by Par-4. AB - Prostate apoptosis response-4 (Par-4) is a unique pro-apoptotic protein that selectively induces apoptosis in cancer cells. Moreover, Par-4 sensitizes cells to the action of diverse apoptotic stimuli and causes tumor regression. This review discusses the prominent structural and functional features of Par-4 and the multiple levels of regulation of its apoptotic function, all of which can be utilized to develop targeted cancer therapy. PMID- 16382047 TI - Morphogenics as a tool for target discovery and drug development. AB - Mutations in DNA mismatch repair (MMR) genes lead to genetically hypermutable cells. Germline mutations in MMR genes in man have been linked to the genetic predisposition to hereditary nonpolyposis colon cancer and a number of other inherited and sporadic malignancies. The ability to modulate the MMR process (referred to as morphogenics) in model systems offers a powerful tool for generating functional diversity in cells and multicellular organisms via the perpetual genomewide accumulation of randomized point and slippage mutation(s). Morphogenics is a platform process that employs a dominant negative MMR gene to create genetic diversity within defined cellular systems and results in a wide range of phenotypes, thus enabling the development and improvement of pharmaceutical products and the discovery of new pharmaceutical targets. Libraries of morphogenics-derived siblings are generated through random mutagenesis from naturally occurring DNA polymerase-induced mutations that occur during DNA replication. Morphogenic cells are screened in high-throughput assays to identify subclones with desired phenotypes for pathway discovery and/or product development. Morphogenics has been successfully applied to a wide range of hosts, including mammalian cells, transgenic mice, plants, yeast, and bacteria. Manipulation of these systems via morphogenics has led to the discovery of novel disease-associated phenotypes in targeted model systems. Moreover, morphogenics has been successfully applied to antibody-producing cell lines to yield subclones producing antibodies with enhanced binding affinities for therapeutic use, as well as to derive subclones with enhanced titers that are suitable for scaleable manufacturing. The selective manipulation of the MMR process via morphogenics is a platform technology that offers many advantages for the discovery of druggable targets, as well as for the development of novel pharmaceutical products. PMID- 16382048 TI - A comprehensive strategy to combat colon cancer targeting the adenomatous polyposis coli tumor suppressor gene. AB - Somatic cells in the majority of colorectal polyps and cancers contain mutations/deletions in the adenomatous polyposis coli (APC) tumor suppressor gene. APC is involved in normal intestinal development and acts to influence a variety of cellular processes. Loss of APC function leads to intestinal neoplasia in both mice and humans. APC influences expression of specific genes, including the c-Myc oncogene, which functions as a transcriptional activator. Loss of APC function leads to alterations in c-Myc-regulated genes including ornithine decarboxylase (ODC), the first enzyme in polyamine synthesis. A single nucleotide polymorphism (SNP) in the ODC promoter affecting c-Myc-dependent expression has been associated with risk of colorectal and other cancers. Pharmaceuticals that target structural features of the c-Myc promoter, and suppress expression of c Myc and other genes regulated by similar promoter elements, are being developed as potential colorectal cancer chemotherapies. Difluoromethylornithine (DFMO), a selective inhibitor of ODC, is under clinical evaluation as a colorectal cancer chemopreventive agent. APC and APC-dependent genes, such as c-Myc and ODC, may be useful as genetic markers of risk and as targets for chemoprevention and therapy for colorectal cancer. PMID- 16382049 TI - External imaging of CCND1, MYC, and KRAS oncogene mRNAs with tumor-targeted radionuclide-PNA-peptide chimeras. AB - In 2005, breast cancer will kill approximately 40,410 women in the U.S., and pancreatic cancer will kill approximately 31,800 men and women in the U.S. Clinical examination and mammography, the currently accepted breast cancer screening methods, miss almost half of breast cancers in women younger than 40 years, approximately one-quarter of cancers in women aged 40-49 years, and one fifth of cancers in women over 50 years old. Pancreatic cancer progresses rapidly, with only 1% of patients surviving more than 5 years after diagnosis. However, if the disease is diagnosed when it is localized, the 5-year survival is approximately 20%. It would be beneficial to detect breast cancer and pancreatic cancer at the earliest possible stage, when multimodal therapy with surgery, radiotherapy, and chemotherapy have the greatest chance of prolonging survival. Human estrogen receptor-positive breast cancer cells typically display elevated levels of Myc protein due to overexpression of MYC mRNA, elevated cyclin D1 protein due to overexpression of CCND1 mRNA, and elevated insulin-like growth factor 1 receptor (IGF1R) due to overexpression of IGF1R mRNA. We hypothesized that scintigraphic detection of MYC or CCND1 peptide nucleic acid (PNA) probes with an IGF1 peptide loop on the C-terminus, and a Tc-99m-chelator peptide on the N-terminus, could measure levels of MYC or CCND1 mRNA noninvasively in human IGF1R-overexpressing MCF7 breast cancer xenografts in immunocompromised mice. Similarly, human pancreatic cancer cells typically display elevated levels of KRAS mRNA and elevated IGF1R. Hence, we also hypothesized that a KRAS Tc-99m chelator PNA-peptide probe could detect overexpression of KRAS mRNA in pancreatic cancer xenografts by scintigraphic imaging, or by positron emission tomography (PET) with a KRAS Cu-64-chelator PNA-peptide. Human MCF7 breast cancer xenografts in immunocompromised mice were imaged scintigraphically 4-24 h after tail-vein administration of MYC or CCND1 Tc-99m-chelator PNA-peptides, but not after administration of mismatch controls. Similarly, human Panc-1 pancreatic cancer cells xenografts were imaged scintigraphically 4 and 24 h after tail-vein administration of a KRAS Tc-99m-chelator PNA-peptide, and AsPC1 xenografts were imaged by PET 4 and 24 h after tail-vein adminstration of a KRAS Cu-64-chelator PNA-peptide. The radioprobes distributed normally to the kidneys, livers, tumors, and other tissues. External molecular imaging of oncogene mRNAs in solid tumors with radiolabel-PNA-peptide chimeras might in the future provide additional genetic characterization of pre-invasive and invasive breast cancers. PMID- 16382050 TI - Functional validation of genes implicated in lymphomagenesis: an in vivo selection assay using a Myc-induced B-cell tumor. AB - The involvement of the c-Myc transcription factor in neoplastic transformation is well documented. However, which of its numerous target genes are crucial for tumorigenesis remains a frequently contested issue. We have recently established a non-transgenic murine model for B-cell lymphoma based on neoplastic conversion of p53-null bone marrow cells by conditionally active Myc. Using this model, we have identified a number of genes whose expression levels are affected by Myc during B-lymphomagenesis. Here we discuss their possible roles in neoplastic processes and describe an experimental approach allowing in vivo validation of these roles. We demonstrate that lymphoma cells overexpressing one of the Myc targets, the interleukin-10 receptor gene, have a very strong selective advantage over low IL10R expressors. Furthermore, Mcl1, a presumptive IL10R effector, also confers selective advantages when overexpressed in Myc-transformed hematopoietic cells. Thus, both IL10R and Mcl1 might be amenable to therapeutic interventions, and new targets can be identified and validated using the selection approach. PMID- 16382051 TI - The proteasome inhibitor bortezomib (Velcade) sensitizes some human tumor cells to Apo2L/TRAIL-mediated apoptosis. AB - On testing a panel of different human cancer cell lines, we observed that the proteasome inhibitor bortezomib could dramatically sensitize some lines to the apoptotic effects of Apo2L/TRAIL. Certain renal, colon, or breast tumor cell lines were dramatically sensitized, whereas other tumor lines from the same tissue of origin remained resistant. This sensitization did not correlate with either the p53 status of the individual tumor cell lines or their intrinsic sensitivity to Apo2L/TRAIL. Colon cancer cell lines lacking p53 or Bax were sensitized by bortezomib, suggesting that neither p53 nor Bax levels were crucial for sensitization. Although the molecular basis of bortezomib sensitization of tumor cells to Apo2L/TRAIL remains to be determined, this combination can have an enhanced apoptotic effect over either agent alone for certain human cancer cells. PMID- 16382052 TI - The role of tuberin in cellular differentiation: are B-Raf and MAPK involved? AB - Tuberous sclerosis complex (TSC) is a tumor suppressor gene syndrome whose manifestations can include seizures, mental retardation, autism, and tumors in the brain, retina, kidney, heart, and skin. The products of the TSC1 and TSC2 genes, hamartin and tuberin, respectively, heterodimerize and inhibit the mammalian target of rapamycin (mTOR). This review focuses on the genetic and biochemical basis of the renal and pulmonary manifestations of TSC, angiomyolipomas, and lymphangiomyomatosis, respectively. Genetic analyses of sporadic angiomyolipomas revealed that all three components (smooth muscle, vessels, and fat) derive from a common progenitor cell, indicating the ability of cells lacking tuberin to differentiate into multiple lineages. Other genetic studies showed that the benign smooth muscle cells of pulmonary lymphangiomyomatosis have the ability to migrate to other organs. These findings suggest that tuberin and hamartin play a role in the regulation of cellular migration and differentiation. We have found that tuberin activates B-Raf kinase and p42/44 MAPK and that cells lacking tuberin have low levels of B-Raf activity. We hypothesize that aberrant B-Raf activity in angiomyolipomas leads to abnormal cellular differentiation and migration. PMID- 16382053 TI - A role for Hath1, a bHLH transcription factor, in colon adenocarcinoma. AB - A significant reduction or loss of goblet cells is often observed in clinical samples of colon adenocarcinomas, which is the predominant form of colon carcinoma. Mice lacking Math1, a bHLH transcription factor downstream of the Notch signaling pathway, demonstrates that Math1 is necessary for cell fate determination of the intestinal secretory cells, including goblet cells. Examination of Hath1, the human orthologue of Math1, expression in multiple colon tumor samples and colon cancer cell lines reveals a dramatic decrease in Hath1 expression in colon tumor samples and colon cancer cell lines. Hath1 expression in the HT29 colon cancer cell line can significantly inhibit its proliferation and anchorage-independent growth both in vitro and in vivo. At the molecular level, Hath1 may regulate the expression of MUC2, a mucin secreted by goblet cells, and Hath1 may also be a novel factor normally repressed as a consequence of activation of the Wnt signaling pathway, which has been clearly implicated in colon tumorigenesis. PMID- 16382054 TI - Alterations in DNA repair gene expression under hypoxia: elucidating the mechanisms of hypoxia-induced genetic instability. AB - Hypoxia is a common feature of solid tumors and is associated with genetic instability and tumor progression. It has been shown previously that alterations in the expression of DNA repair genes in response to hypoxic stress may account for a proportion of such genetic instability. Here, we demonstrate that the expression of RAD51, a critical mediator of homologous recombination (HR), is repressed by hypoxia in numerous cell lines derived from a wide range of tissues. Repression of this gene by hypoxia occurs in a cell cycle- and hypoxia-inducible factor (HIF)-independent manner, and decreased RAD51 expression was observed to persist during the post-hypoxic period. In addition, decreases in Rad51 expression were correlated with functional impairments in HR repair in hypoxic and post-hypoxic cells. Based on these data, we propose a novel mechanism of hypoxia-induced genetic instability via suppression of the HR pathway in cancer cells within the tumor microenvironment. PMID- 16382055 TI - Images in clinical medicine. Buckshot ingestion. PMID- 16382056 TI - A prescription for a modern Medicare program. PMID- 16382057 TI - Promise and perils for patients and physicians. PMID- 16382058 TI - Benefits and consequences for the poor and the disabled. PMID- 16382059 TI - A beneficial side effect of the Medicare drug benefit. PMID- 16382060 TI - Preventing stroke in sickle cell anemia. PMID- 16382061 TI - A comparison of letrozole and tamoxifen in postmenopausal women with early breast cancer. AB - BACKGROUND: The aromatase inhibitor letrozole is a more effective treatment for metastatic breast cancer and more effective in the neoadjuvant setting than tamoxifen. We compared letrozole with tamoxifen as adjuvant treatment for steroid hormone-receptor-positive breast cancer in postmenopausal women. METHODS: The Breast International Group (BIG) 1-98 study is a randomized, phase 3, double blind trial that compared five years of treatment with various adjuvant endocrine therapy regimens in postmenopausal women with hormone-receptor-positive breast cancer: letrozole, letrozole followed by tamoxifen, tamoxifen, and tamoxifen followed by letrozole. This analysis compares the two groups assigned to receive letrozole initially with the two groups assigned to receive tamoxifen initially; events and follow-up in the sequential-treatment groups were included up to the time that treatments were switched. RESULTS: A total of 8010 women with data that could be assessed were enrolled, 4003 in the letrozole group and 4007 in the tamoxifen group. After a median follow-up of 25.8 months, 351 events had occurred in the letrozole group and 428 events in the tamoxifen group, with five-year disease-free survival estimates of 84.0 percent and 81.4 percent, respectively. As compared with tamoxifen, letrozole significantly reduced the risk of an event ending a period of disease-free survival (hazard ratio, 0.81; 95 percent confidence interval, 0.70 to 0.93; P=0.003), especially the risk of distant recurrence (hazard ratio, 0.73; 95 percent confidence interval, 0.60 to 0.88; P=0.001). Thromboembolism, endometrial cancer, and vaginal bleeding were more common in the tamoxifen group. Women given letrozole had a higher incidence of skeletal and cardiac events and of hypercholesterolemia. CONCLUSIONS: In postmenopausal women with endocrine-responsive breast cancer, adjuvant treatment with letrozole, as compared with tamoxifen, reduced the risk of recurrent disease, especially at distant sites. (ClinicalTrials.gov number, NCT00004205.) PMID- 16382062 TI - Rescue angioplasty after failed thrombolytic therapy for acute myocardial infarction. AB - BACKGROUND: The appropriate treatment for patients in whom reperfusion fails to occur after thrombolytic therapy for acute myocardial infarction remains unclear. There are few data comparing emergency percutaneous coronary intervention (rescue PCI) with conservative care in such patients, and none comparing rescue PCI with repeated thrombolysis. METHODS: We conducted a multicenter trial in the United Kingdom involving 427 patients with ST-segment elevation myocardial infarction in whom reperfusion failed to occur (less than 50 percent ST-segment resolution) within 90 minutes after thrombolytic treatment. The patients were randomly assigned to repeated thrombolysis (142 patients), conservative treatment (141 patients), or rescue PCI (144 patients). The primary end point was a composite of death, reinfarction, stroke, or severe heart failure within six months. RESULTS: The rate of event-free survival among patients treated with rescue PCI was 84.6 percent, as compared with 70.1 percent among those receiving conservative therapy and 68.7 percent among those undergoing repeated thrombolysis (overall P=0.004). The adjusted hazard ratio for the occurrence of the primary end point for repeated thrombolysis versus conservative therapy was 1.09 (95 percent confidence interval, 0.71 to 1.67; P=0.69), as compared with adjusted hazard ratios of 0.43 (95 percent confidence interval, 0.26 to 0.72; P=0.001) for rescue PCI versus repeated thrombolysis and 0.47 (95 percent confidence interval, 0.28 to 0.79; P=0.004) for rescue PCI versus conservative therapy. There were no significant differences in mortality from all causes. Nonfatal bleeding, mostly at the sheath insertion site, was more common with rescue PCI. At six months, 86.2 percent of the rescue-PCI group were free from revascularization, as compared with 77.6 percent of the conservative-therapy group and 74.4 percent of the repeated thrombolysis group (overall P=0.05). CONCLUSIONS: Event-free survival after failed thrombolytic therapy was significantly higher with rescue PCI than with repeated thrombolysis or conservative treatment. Rescue PCI should be considered for patients in whom reperfusion fails to occur after thrombolytic therapy. PMID- 16382063 TI - Discontinuing prophylactic transfusions used to prevent stroke in sickle cell disease. AB - BACKGROUND: Prophylactic transfusion prevents strokes in children with sickle cell anemia who have abnormalities on transcranial Doppler ultrasonographic examination. However, it is not known how long transfusion should be continued in these children. METHODS: We studied children with sickle cell disease who had a high risk of stroke on the basis of a transcranial Doppler screening examination and who had received transfusions for 30 months or longer, during which time the Doppler readings became normal. The children were randomly assigned to continued transfusion or no continued transfusion. Children with severe stenotic lesions on cerebral magnetic resonance angiography were excluded. The composite primary end point was stroke or reversion to a result on Doppler examination indicative of a high risk of stroke. RESULTS: The study was stopped after 79 children of a planned enrollment of 100 underwent randomization. Among the 41 children in the transfusion-halted group, high-risk Doppler results developed in 14 and stroke in 2 others within a mean (+/-SD) of 4.5+/-2.6 months (range, 2.1 to 10.1) of the last transfusion. Neither of these events of the composite end point occurred in the 38 children who continued to receive transfusions. The average of the last two transcranial Doppler results before transfusion was started was the only predictor of the composite end point (P=0.05). CONCLUSIONS: Discontinuation of transfusion for the prevention of stroke in children with sickle cell disease results in a high rate of reversion to abnormal blood-flow velocities on Doppler studies and stroke. (ClinicalTrials.gov number, NCT00006182.) PMID- 16382064 TI - Trial Registration at ClinicalTrials.gov between May and October 2005. AB - BACKGROUND: Clinical trial registration allows interested parties to obtain information about ongoing and completed trials, but there are few data indicating the quality of the information provided during the registration process. We used information in the publicly available ClinicalTrials.gov database to describe patterns of trial registration before and after the implementation by journal editors of a new policy requiring registration as a prerequisite for publication. METHODS: We reviewed ClinicalTrials.gov records to determine patterns of completion of the "Intervention Name" and "Primary Outcome Measure" data fields for trials registered on May 20 and October 11, 2005, and for trials registered during the interval between these two dates, inclusively. RESULTS: During the interval studied, the number of registrations in ClinicalTrials.gov increased by 73 percent from 13,153 to 22,714. The percentage of interventional trials registered by industry with nonspecific Intervention Name entries (attributable to four drug companies) decreased from 10 percent to 2 percent; all other industry and nonindustry records contained specific entries in this field. Of the 2670 studies registered by industry between the two dates, 76 percent provided information in the Primary Outcome Measure field, although these entries varied markedly in their degree of specificity. In the remaining 24 percent of the records, this field was blank. CONCLUSIONS: During the summer of 2005, there were large increases in the number of clinical trial registrations. Overall, the data contained in records were more complete in October than they were in May, but there still is room for substantial improvement. PMID- 16382065 TI - Clinical practice. Acute pulmonary edema. PMID- 16382066 TI - Images in clinical medicine. Acanthosis nigricans. PMID- 16382067 TI - Case records of the Massachusetts General Hospital. Case 40-2005. An 18-year-old man with a one-month history of nontender left mandibular swelling. PMID- 16382068 TI - Aromatase inhibitors--a triumph of translational oncology. PMID- 16382069 TI - Trial registration report card. PMID- 16382070 TI - Registries and registration of clinical trials. PMID- 16382071 TI - Autoantibodies in prostate cancer. PMID- 16382072 TI - Prophylactic thyroidectomy in multiple endocrine neoplasia type 2A. PMID- 16382073 TI - Passive immunization against cytomegalovirus during pregnancy. PMID- 16382074 TI - Staphylococcal sepsis in children. PMID- 16382075 TI - Chaperones and disease. PMID- 16382076 TI - Asymptomatic hyperinsulinemic hypoglycemia after gastric banding. PMID- 16382077 TI - From psychological need satisfaction to intentional behavior: testing a motivational sequence in two behavioral contexts. AB - The present study tested a motivational sequence in which global-level psychological need satisfaction from self-determination theory influenced intentions and behavior directly and indirectly through contextual-level motivation and situational-level decision-making constructs from the theory of planned behavior. Two samples of university students (N = 511) completed measures of global-level psychological need satisfaction, contextual-level autonomous motivation, and situational-level attitudes, subjective norms, perceived behavioral control, intentions, and behavior in two behavioral contexts: exercise and dieting. A structural equation model supported the proposed sequence in both samples. The indirect effect was present for exercise behavior, whereas both direct and indirect effects were found for dieting behavior. Findings independently supported the component theories and provided a comprehensive integrated explanation of volitional behavior. PMID- 16382078 TI - Responses to discrimination: the role of emotion and expectations for emotional regulation. AB - The present study examined the role of emotion in women's perceptions of discrimination and their endorsement of behavioral responses to change the status quo. In an experimental simulation involving a situation of sex discrimination, women (N = 108) were primed to experience a particular emotion (sad, angry, control condition) and were subsequently instructed to either suppress or express (or neither) their emotional responses. Women primed to feel sad and told to suppress their emotions reported the least discrimination, whereas angered women who were permitted to express themselves reported the greatest discrimination. Furthermore, when encouraged to express their emotions, women primed to feel sad were more likely to endorse normative actions to rectify the situation, whereas women induced to feel angry were more likely to endorse collective actions to change the status quo. These findings have implications for the role of emotions and expectations regarding their expression on collective action taking. PMID- 16382079 TI - When group members admit to being conformist: the role of relative intragroup status in conformity self-reports. AB - Five studies examined the hypothesis that people will strategically portray the self as being more group influenced the more junior they feel within the group. Among social psychologists (Study 1), ratings of self-conformity by group members were greater when the status of the participant was low than when it was high. These effects were replicated in Studies 2, 3, and 4 in which relative intragroup status was manipulated. In Study 3, the authors found junior group members described themselves as more conformist than senior members when they were addressing an ingroup audience, but when they were addressing an outgroup audience the effect disappeared. Furthermore, junior members (but not senior members) rated themselves as more conformist when they were led to believe their responses were public than when responses were private (Study 5). The discussion focuses on the strategic processes underlying low-status group members' self reports of group influence and the functional role of conformity in groups. PMID- 16382080 TI - Belief in supernatural agents in the face of death. AB - Four studies examined whether awareness of mortality intensifies belief in supernatural agents among North Americans. In Studies 1 and 2, mortality salience led to more religiosity, stronger belief in God, and in divine intervention. In Studies 3 and 4, mortality salience increased supernatural agent beliefs even when supernatural agency was presented in a culturally alien context (divine Buddha in Study 3, Shamanic spirits in Study 4). The latter effects occurred primarily among the religiously affiliated, who were predominantly Christian. Implications for the role of supernatural agent beliefs in assuaging mortality concerns are discussed. PMID- 16382081 TI - Playing dice with criminal sentences: the influence of irrelevant anchors on experts' judicial decision making. AB - Judicial sentencing decisions should be guided by facts, not by chance. The present research however demonstrates that the sentencing decisions of experienced legal professionals are influenced by irrelevant sentencing demands even if they are blatantly determined at random. Participating legal experts anchored their sentencing decisions on a given sentencing demand and assimilated toward it even if this demand came from an irrelevant source (Study 1), they were informed that this demand was randomly determined (Study 2), or they randomly determined this demand themselves by throwing dice (Study 3). Expertise and experience did not reduce this effect. This sentencing bias appears to be produced by a selective increase in the accessibility of arguments that are consistent with the random sentencing demand: The accessibility of incriminating arguments was higher if participants were confronted with a high rather than a low anchor (Study 4). Practical and theoretical implications of this research are discussed. PMID- 16382082 TI - Working models of attachment and attribution processes in intimate relationships. AB - Two studies examined the link between working models of attachment and social construal processes in romantic relationships. In Study 1, individuals high in attachment-related anxiety responded to hypothetical partner transgressions by endorsing relationship-threatening attributions, experiencing emotional distress, and endorsing behavioral intentions that were likely to result in conflict. These effects emerged after controlling for pessimistic explanatory style, depressed mood, and self-esteem. In addition, the association between anxiety and emotional distress was mediated by attributions and attachment-related needs. In Study 2, anxious individuals endorsed relationship-threatening attributions for their partner's transgressions but less so for their partner's positive behaviors, and these effects occurred primarily among those in unhappy relationships. In contrast, avoidant individuals endorsed pessimistic attributions for their partner's positive behavior but less so for their partner's transgressions, and these effects occurred regardless of their level of relationship satisfaction. PMID- 16382083 TI - Passion and psychological adjustment: a test of the person-environment fit hypothesis. AB - Passion represents a strong inclination toward an activity that is important, liked, and in which significant time is invested. Although a harmonious passion is well integrated in one's identity and is emitted willingly, obsessive passion is not well integrated and is emitted out of internal pressure. This study tested for the presence of a Passion x Environment fit interaction with respect to psychological adjustment. Elite hockey players (N = 233) who tried out for a team in a highly competitive league participated in this short-term longitudinal study. As hypothesized, being selected by the highly competitive leagues led to higher psychological adjustment than not being selected by such leagues. Two months later, an interaction revealed that among athletes who were playing in highly competitive leagues, obsessively passionate athletes reported higher psychological adjustment than did harmonious athletes. Conversely, among athletes playing in less competitive leagues, harmonious athletes reported higher psychological adjustment than did obsessive athletes. PMID- 16382084 TI - Recategorization and subgroup identification: predicting and preventing threats from common ingroups. AB - Much work has supported the idea that recategorization of ingroups and outgroups into a superordinate category can have beneficial effects for intergroup relations. Recently, however, increases in bias following recategorization have been observed in some contexts. It is argued that such unwanted consequences of recategorization will only be apparent for perceivers who are highly committed to their ingroup subgroups. In Experiments 1 to 3, the authors observed, on both explicit and implicit measures, that an increase in bias following recategorization occurred only for high subgroup identifiers. In Experiment 4, it was found that maintaining the salience of subgroups within a recategorized superordinate group averted this increase in bias for high identifiers and led overall to the lowest levels of bias. These findings are discussed in the context of recent work on the Common Ingroup Identity Model. PMID- 16382085 TI - The carrot and the stick: affective commitment and acceptance anxiety as motives for discretionary group efforts by respected and disrespected group members. AB - Previous research has demonstrated that intragroup respect can strengthen people's commitment to the group and encourage them to exert themselves on behalf of it. In the present research, the authors argue that similar behavior can ensue from self-focused concerns when group members are disrespected. Experiment 1 (N = 174) confirms that high respect as well as low respect motivates people to increase their actual discretionary efforts on behalf of the group. These findings were replicated and extended in Experiment 2 (N = 138), where it was established that enhanced efforts only emerge when people consider the way they are evaluated by others as diagnostic for their position in the group. In addition, it is demonstrated that whereas the efforts of respected people were primarily motivated by affective commitment to the group (group-focused concerns), the behavior of disrespected people was driven by anxiety about their acceptance into the group (self-focused concerns). PMID- 16382086 TI - Push-you-pull-you: the boundaried self in close relationships. AB - Based on Amoebic Self Theory, the authors propose that the salience of different threats to the self affects the extent to which an intimate relationship partner is pushed away (excluded) or pulled closer (included). When social threat is salient among persons in relationships, it is hypothesized that partners will attempt to defuse the resulting sense of interpersonal vulnerability: offending partners may be pushed away, whereas offenders themselves may draw closer. When spatial-symbolic threat is salient and the relationship's capacity to function as an identity marker is jeopardized, it is hypothesized that the partner- regardless of his or her role--will be pulled closer to maintain the perception that the relationship is secure. Self-report responses to hypothetical scenarios and perceptions of behaviors during a role-play were generally consistent with these hypotheses, suggesting that both an intimate partner and the relationship with that partner can be incorporated into the self. PMID- 16382087 TI - The relations between family environment and violence exposure among youth: findings from the national survey of adolescents. AB - A national household probability sample of 4,023 adolescents (ages 12 to 17) completed telephone interviews assessing demographics, adverse family environment, and violence exposure. Logistic regressions examined relations among family environment and each violence exposure type, controlling for demographics and other violence exposures. Relationships between family environment and violence exposure varied, depending on type of violence reported, most notably between intrafamilial versus extrafamilial violence. After controlling for family environment, exposure to one violence type significantly increased the likelihood of other violence exposures. Family substance use and not always living with a natural parent were significantly associated with all three types of violence exposure. Findings indicate that clinical assessments should include a thorough evaluation of family environment and violence exposure and also highlight the need for treatment to focus on the adolescent and broader family unit. Future research is needed to further examine these complex interrelationships and their associations with adolescent outcomes. PMID- 16382088 TI - Ethnicity in child maltreatment research: a replication of Behl et al.'s content analysis. AB - This study examines the use of ethnicity in 489 empirical research articles published in three major child maltreatment specialty journals from 1999 to 2002. Of the American samples, 12.5% focus on ethnicity, 76.2% report the ethnic composition of participants, and 33.8% use ethnicity of participants in analyses. Ethnicity has a significant effect in 52.3% of articles in which it was used in analyses, suggesting its importance as a variable in a wide range of studies. African Americans and Native Americans are underrepresented in research samples. These findings indicate more attention to ethnicity in American research than Behl, Crouch, May, Valente, and Conyngham's 2001 study might suggest but also highlight the need for continued expansion in focusing on, reporting, and using ethnicity in research. PMID- 16382089 TI - The role of men in chronic supervisory neglect. AB - This study investigates whether parents' child care demand and resources, their capacity to provide adequate supervision, and their understanding of the supervision problem predict chronic supervisory neglect. A case-comparison design was used to compare families who had one isolated incident of supervisory neglect, who were involved with child protective services (CPS) because of a persistent supervision problem (2 years or less), and who were involved with CPS because of a chronic supervision problem (more than 2 years). When the mother's partner was not the father of her children or had a drug, alcohol, or mental health challenge, and when no one understood that there was a supervision problem or took responsibility for it, the problem was more likely to persist or become chronic. Therefore, when predicting whether a family will continue to provide inadequate supervision, it is important to also assess the mother's partner. PMID- 16382090 TI - Weekly problems scales: instruments for sexually abused youth and their nonoffending parents in treatment. AB - This study's purpose was to determine if efficient measures could be created to assess multiple problematic behaviors identified in youth who were sexually abused and in treatment. Because of the lack of easily administered brief instruments that assess multiple domains of interest in this population, complementary parent and child assessment measures were developed. The Weekly Problems Scale-Child Version (WPSC) and the Weekly Problems Scale-Parent Version (WPS-P) were created to monitor the weekly progress of the child and family in treatment and focus specifically on common areas of difficulties in this population. Exploratory factor analysis was conducted to assist in identifying the number of underlying dimensions in the scales. Results indicate that the WPS C and WPS-P demonstrate adequate internal consistency, temporal stability, and construct validity. The WPS-C and WPSP display significant promise as research and clinical assessment tools for use with youth who are sexually abused and their nonoffending parents in treatment. PMID- 16382091 TI - Ecological influences on the sequelae of child maltreatment: a review of the literature. AB - Numerous studies indicate that child maltreatment increases the risk for the development of internalizing and externalizing behavior problems. Great variations in outcome, however, have been noted among victims of maltreatment. From an ecological perspective, this review examines how the effects of maltreatment may be influenced by the contexts in which children develop, including their families, peer groups, schools, and communities. The literature reviewed suggests that contextual factors not only influence the incidence of maltreatment but also may moderate its developmental effects, thereby accounting for some of the heterogeneity in the outcomes associated with abuse and neglect. Closer examination of the influence contextual factors exert on the psychosocial sequelae of maltreatment will better inform the interventions, treatments, and public policies directed toward the maltreated population. Methodological considerations for conducting research in this area are also discussed. PMID- 16382092 TI - Relations of parental report and observation of parenting to maltreatment history. AB - Parenting assessments (the Parent-Child Conflict Tactics Scale, CTSPC; and a mother-child observation) were examined for their ability to identify mothers with a history of physically abusing or neglecting their child. Participants were mothers of 139 children (age 3 to 6 years; 58 with a history of maltreatment). Mothers with a history of maltreatment reported higher scores on the Neglect, Nonviolent Discipline, and Psychological Aggression subscales of the CTSPC. These group differences, however, were limited to mothers who acknowledged a history of maltreatment, as mothers who concealed their maltreatment history rated themselves similar to controls. Observation of parental behaviors during a brief, nonstressful task did not discriminate mothers who maltreated from mothers who did not maltreat. The findings suggest that parental report using the CTSPC may be useful in assessing parenting behaviors among mothers with a history of maltreatment, although socially desirable responding is a significant problem. PMID- 16382093 TI - Report of the APSAC task force on attachment therapy, reactive attachment disorder, and attachment problems. AB - Although the term attachment disorder is ambiguous, attachment therapies are increasingly used with children who are maltreated, particularly those in foster care or adoptive homes. Some children described as having attachment disorders show extreme disturbances. The needs of these children and their caretakers are real. How to meet their needs is less clear. A number of attachment-based treatment and parenting approaches purport to help children described as attachment disordered. Attachment therapy is a young and diverse field, and the benefits and risks of many treatments remain scientifically undetermined. Controversies have arisen about potentially harmful attachment therapy techniques used by a subset of attachment therapists. In this report, the Task Force reviews the controversy and makes recommendations for assessment, treatment, and practices. The report reflects American Professional Society on the Abuse of Children's (APSAC) position and also was endorsed by the American Psychological Association's Division 37 and the Division 37 Section on Child Maltreatment. PMID- 16382094 TI - The early intervention foster care program: a glass half full. PMID- 16382095 TI - "The effects of early prevention programs for families with young children at risk for physical child abuse and neglect: a meta-analysis" by Liesl Geeraert and coworkers. PMID- 16382097 TI - Overview of molecular relationships in the voltage-gated ion channel superfamily. PMID- 16382098 TI - International Union of Pharmacology. XLVII. Nomenclature and structure-function relationships of voltage-gated sodium channels. AB - The family of voltage-gated sodium channels initiates action potentials in all types of excitable cells. Nine members of the voltage-gated sodium channel family have been characterized in mammals, and a 10th member has been recognized as a related protein. These distinct sodium channels have similar structural and functional properties, but they initiate action potentials in different cell types and have distinct regulatory and pharmacological properties. This article presents the molecular relationships and physiological roles of these sodium channel proteins and provides comprehensive information on their molecular, genetic, physiological, and pharmacological properties. PMID- 16382099 TI - International Union of Pharmacology. XLVIII. Nomenclature and structure-function relationships of voltage-gated calcium channels. AB - The family of voltage-gated calcium channels serves as the key transducers of cell surface membrane potential changes into local intracellular calcium transients that initiate many different physiological events. There are 10 members of the voltage-gated calcium channel family that have been characterized in mammals, and they serve distinct roles in cellular signal transduction. This article presents the molecular relationships and physiological functions of these calcium channel proteins and provides comprehensive information on their molecular, genetic, physiological, and pharmacological properties. PMID- 16382100 TI - International Union of Pharmacology. XLIX. Nomenclature and structure-function relationships of transient receptor potential channels. PMID- 16382101 TI - International Union of Pharmacology. L. Nomenclature and structure-function relationships of CatSper and two-pore channels. PMID- 16382102 TI - International Union of Pharmacology. LI. Nomenclature and structure-function relationships of cyclic nucleotide-regulated channels. PMID- 16382103 TI - International Union of Pharmacology. LII. Nomenclature and molecular relationships of calcium-activated potassium channels. PMID- 16382104 TI - International Union of Pharmacology. LIII. Nomenclature and molecular relationships of voltage-gated potassium channels. PMID- 16382105 TI - International Union of Pharmacology. LIV. Nomenclature and molecular relationships of inwardly rectifying potassium channels. PMID- 16382106 TI - International Union of Pharmacology. LV. Nomenclature and molecular relationships of two-P potassium channels. PMID- 16382107 TI - International Union of Pharmacology. LVI. Ghrelin receptor nomenclature, distribution, and function. AB - Ghrelin is a 28-amino acid peptide originally isolated from rat stomach and is cleaved from a 117-amino acid precursor. The sequence of the mature peptide from rats and mice differs by two amino acids from that of human ghrelin. Alternative splicing of the ghrelin gene transcript can result in the translation of a second biologically active peptide, des-Gln14-ghrelin. Both peptides have a unique post translational modification, octanoylation of Ser3, which is essential for the binding to receptors in hypothalamus and pituitary and stimulating the release of growth hormone from the pituitary. The growth hormone secretagogue receptor (GHS R1a, Swiss-Prot code Q92847, LocusLink ID 2693), a rhodopsin-like seven transmembrane spanning G protein-coupled receptors belonging to Family A, was cloned in 1996 from the pituitary and hypothalamus and shown to be the target of growth hormone secretagogues (GHS), a class of synthetic peptide and nonpeptide compounds causing growth hormone release from the anterior pituitary. In 1999, ghrelin was identified as the endogenous cognate ligand for this receptor. The purpose of this review is to propose an official International Union of Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC IUPHAR) nomenclature designating GHS-R1a as the ghrelin receptor to follow the convention of naming receptors after the endogenous agonist, abbreviated where necessary to GRLN. PMID- 16382108 TI - The evolution of iron chelators for the treatment of iron overload disease and cancer. AB - The evolution of iron chelators from a range of primordial siderophores and aromatic heterocyclic ligands has lead to the formation of a new generation of potent and efficient iron chelators. For example, various siderophore analogs and synthetic ligands, including ICL670A [4-[3,5-bis-(hydroxyphenyl)-1,2,4-triazol-1 yl]-benzoic acid], 4'-hydroxydesazadesferrithiocin, and Triapine, have been developed from predecessors and illustrate potent iron-mobilizing or antineoplastic activities. This review focuses on the evolution of iron chelators from initial lead compounds through to the development of novel chelating agents, many of which show great potential to be clinically applied in the treatment of iron overload disease and cancer. PMID- 16382109 TI - Carbon monoxide: endogenous production, physiological functions, and pharmacological applications. AB - Over the last decade, studies have unraveled many aspects of endogenous production and physiological functions of carbon monoxide (CO). The majority of endogenous CO is produced in a reaction catalyzed by the enzyme heme oxygenase (HO). Inducible HO (HO-1) and constitutive HO (HO-2) are mostly recognized for their roles in the oxidation of heme and production of CO and biliverdin, whereas the biological function of the third HO isoform, HO-3, is still unclear. The tissue type-specific distribution of these HO isoforms is largely linked to the specific biological actions of CO on different systems. CO functions as a signaling molecule in the neuronal system, involving the regulation of neurotransmitters and neuropeptide release, learning and memory, and odor response adaptation and many other neuronal activities. The vasorelaxant property and cardiac protection effect of CO have been documented. A plethora of studies have also shown the importance of the roles of CO in the immune, respiratory, reproductive, gastrointestinal, kidney, and liver systems. Our understanding of the cellular and molecular mechanisms that regulate the production and mediate the physiological actions of CO has greatly advanced. Many diseases, including neurodegenerations, hypertension, heart failure, and inflammation, have been linked to the abnormality in CO metabolism and function. Enhancement of endogenous CO production and direct delivery of exogenous CO have found their applications in many health research fields and clinical settings. Future studies will further clarify the gasotransmitter role of CO, provide insight into the pathogenic mechanisms of many CO abnormality-related diseases, and pave the way for innovative preventive and therapeutic strategies based on the physiologic effects of CO. PMID- 16382110 TI - Aneurysmal bone cyst: a neoplasm driven by upregulation of the USP6 oncogene. PMID- 16382111 TI - Flare in neuropathy following rituximab therapy for Waldenstrom's macroglobulinemia. PMID- 16382112 TI - Comparison of CA-125 and standard definitions of progression of ovarian cancer in the intergroup trial of cisplatin and paclitaxel versus cisplatin and cyclophosphamide. AB - PURPOSE: A definition for progression of ovarian cancer has been proposed based on either a confirmed doubling of CA-125 levels from the upper limit of normal or from the nadir level if levels are persistently elevated. Retrospectively, we determined whether the use of this CA-125 definition in a randomized trial would have shown the same magnitude of difference between the treatment arms as was shown when the standard progression definition was used. PATIENTS AND METHODS: A retrospective analysis was performed on 680 patients in the Taxol Intergroup Trial with advanced epithelial ovarian carcinoma, of whom 628 were assessable according to CA-125. The date of progression according to clinical or radiologic criteria was compared with the date of progression according to CA-125. RESULTS: Of the 628 patients assessable for both definitions, 556 clinical or radiologic progressions were determined compared with 389 according to the CA-125 definition. There was a highly significant difference in the hazard of progression between the paclitaxel and cisplatin arm (TP) compared with the cyclophosphamide and cisplatin arm (CP) when either standard or CA-125 criteria were used to define progression (standard, P = .002; CA-125, P = .011). The hazard ratio of TP/CP over time was similar when comparing the different methods of defining progression. CONCLUSION: The results of this analysis show that the magnitude of the therapeutic benefit was similar whether CA-125 or standard criteria were used to define progression. PMID- 16382113 TI - Early identification of resistance to first-line single-agent methotrexate in patients with persistent trophoblastic disease. AB - PURPOSE: A generally accepted definition for resistance to first-line single agent chemotherapy for persistent trophoblastic disease (PTD) is lacking. In the present study, a normogram for serum human chorionic gonadotropin (hCG) from patients with normalization of serum hCG after first-line single-agent chemotherapy for PTD was constructed to identify patients resistant to this chemotherapy. PATIENTS AND METHODS: Between 1987 and 2004, data from 2,132 patients were registered at the Dutch Central Registry for Hydatidiform Moles. A normal serum hCG regression corridor was constructed for 79 patients with low risk PTD who were cured by single-agent methotrexate (MTX) chemotherapy (control group). Another group of 29 patients with low-risk PTD needed additional alternative therapies (dactinomycin and multiagent chemotherapy) for failure of serum hCG to normalize with single-agent chemotherapy (study group). RESULTS: Serum hCG measurement preceding the fourth and sixth single-agent chemotherapy course proved to have excellent diagnostic accuracy for identifying resistance to single-agent chemotherapy, with an area under the curve (AUC) for receiver operating characteristic curve analysis of 0.949 and 0.975, respectively. At 97.5% specificity, serum hCG measurements after 7 weeks showed 50% sensitivity. CONCLUSION: In the largest study to date, we describe the regression of serum hCG levels in patients with low-risk PTD successfully treated with MTX. At high specificity, hCG levels in the first few courses of MTX can identify half the number of patients who are extremely likely to need alternative chemotherapy to cure their disease and for whom further treatment with single-agent chemotherapy will be ineffective. PMID- 16382114 TI - First-line single agent treatment with gefitinib in patients with advanced non small-cell lung cancer: a phase II study. AB - PURPOSE: We conducted a phase II study of single agent treatment with gefitinib in chemotherapy-naive patients with advanced non-small-cell lung cancer (NSCLC) to assess its efficacy and toxicity. PATIENTS AND METHODS: Patients received 250 mg doses of gefitinib daily. Administration of gefitinib was terminated if partial response (PR) was not achieved within 8 weeks or if tumor reduction was not observed within 4 weeks. In these cases, platinum-based doublet chemotherapy was given as a salvage treatment. We evaluated mutation status of the epidermal growth factor receptor (EGFR) gene in cases with available tumor samples. RESULTS: Forty-two patients were enrolled between March and November 2003, with 40 of these patients being eligible. The response rate was 30% (95% CI, 17% to 47%). The most common toxicity included grade 1 or 2 acne-like rash (50%) and grade 1 diarrhea (18%). Grade 2 or 3 hepatic toxicity was observed in 8% of patients. Four patients developed grade 5 interstitial lung disease (ILD). Thirty patients received second-line chemotherapy. Median survival time was 13.9 months (95% CI, 9.1 to 18.7 months), and the 1-year survival rate was 55%. Tumor samples were available in 13 patients, including four cases of PR, six cases of stable disease, and three cases of progressive disease. EGFR mutations (deletions in exon 19 or point mutations [L858R or E746V]) were detected in four tumor tissues. All four patients with EGFR mutation achieved PR with gefitinib treatment. CONCLUSION: Single agent treatment with gefitinib is active in chemotherapy-naive patients with advanced NSCLC, but produces unacceptably frequent ILD in the Japanese population. PMID- 16382115 TI - Neuroendocrine neoplasms of the lung: a prognostic spectrum. AB - PURPOSE: Neuroendocrine (NE) tumors of the lung include typical carcinoid (TC), atypical carcinoid (AC), large-cell NE carcinoma (LCNEC), and small-cell lung carcinoma (SCLC). Their clinicopathologic profiles and relative grade of malignancy have not been defined. PATIENTS AND METHODS: From 10 Japanese institutes, 383 surgically resected pulmonary NE tumors were collected. The histologic diagnosis was determined by the consensus of a pathology panel consisting of six expert pathologists as TC, AC, LCNEC, or SCLC on the basis of the WHO classification, and its relationship to clinicopathologic profiles was analyzed. RESULTS: Of the 383 tumors, 18 were excluded because of an improper specimen. The pathology panel reviewed the remaining 366 tumors, and a diagnosis of NE tumor was made in 318 patients (87.4%); 55 patients had TC, nine had AC, 141 had LCNEC, and 113 had SCLC. The 5-year survival rates of patients with all stages were as follows: 96.2% for TC, 77.8% for AC, 40.3% for LCNEC, and 35.7% for SCLC. There was significant prognostic difference between TC and AC as well as between AC and LCNEC+SCLC. However, there was no difference between LCNEC and SCLC, and their survival curves were superimposed. The multivariate analysis indicated that histologic type, completeness of resection, symptoms, nodal involvement, and age were significantly prognostic. CONCLUSION: The grade of malignancy of NE tumors was upgraded in the following order: TC, AC, LCNEC, and SCLC. No prognostic difference was noted between LCNEC and SCLC. The high-grade NE histology uniformly indicated poor prognosis regardless of its histologic type. PMID- 16382116 TI - Patterns of care in early-stage breast cancer survivors in the first year after cessation of active treatment. AB - PURPOSE: Patterns of health care use have not been well described for breast cancer survivors. The purpose of this study was to describe the health service use in a survivor cohort. PATIENTS AND METHODS: Women with stage I or II breast cancer were recruited (n = 558) after primary treatment for a multicenter, randomized trial of psychoeducational interventions for facilitating transition to survivorship; 418 women completed the study. Participants completed calendar diaries detailing health care use for 1 year after treatment. Services were coded using Current Procedural Terminology-Fourth Edition codes; costs were estimated using year 2000 Medicare reimbursements. RESULTS: Health care use diary data were available for 391 women (70% of the sample). On average, these survivors reported 30 episodes of health service use in the year after treatment. Total annual costs of care averaged more than 1,800 dollars per survivor; medical office visits were the major component of costs. Type of cancer treatment, depression, and physical function and comorbid illness were independent predictors of the costs of services. There were geographic variations in initial local treatment patterns and in post-treatment costs. Notably, all women should have received surveillance mammography in the time period, but only 61.9% did so; the odds of mammogram receipt were higher for women who had a lumpectomy (v mastectomy) and women who were white (v nonwhite). CONCLUSION: Use of health services is frequent and intensive in the first year after treatment for breast cancer. Despite frequent contact with the health care system, there is room for improvement in providing guideline-suggested surveillance mammography for survivors. PMID- 16382117 TI - Factors related to underuse of surveillance mammography among breast cancer survivors. AB - PURPOSE: Many older breast cancer survivors do not undergo annual mammography despite guideline recommendations. We identified factors associated with underuse of surveillance mammography and examined whether variation was explained by differences in follow-up care. PATIENTS AND METHODS: We used Surveillance, Epidemiology, and End Results-Medicare data to identify a population-based sample of 44,511 women fee-for-service Medicare enrollees aged > or = 65 years who were diagnosed with stage I or II breast cancer in 1992 to 1999 who underwent primary surgical therapy. We assessed factors associated with mammography during months 7 to 18, 19 to 30, and 31 to 42 after breast cancer diagnosis using repeated measures logistic regression; and we examined whether follow-up care with providers of various specialties explained variation in mammography use. RESULTS: Only three quarters of women (77.6%) underwent mammography during months 7 to 18 after diagnosis, and only 56.7% had mammography yearly over 3 years. In multivariable analyses, women who were older, black, unmarried, and living in certain regions were less likely than other women to undergo surveillance mammography (all P < .05). Patients with more visits and patients who continued to see a medical oncologist, radiation oncologist, or surgeon were most likely to have mammograms (P < .001); however, adjusting for visits with providers did not explain the lower mammography rates based on age, race, marital status, and geographic region. CONCLUSION: Many elderly breast cancer survivors do not undergo annual surveillance mammography, particularly women who are older, black, and unmarried, and this underuse was not explained by access to follow-up care. New strategies are needed to increase use of surveillance mammography and decrease variations based on nonclinical factors that are likely unrelated to appropriateness of medical care. PMID- 16382118 TI - Psychological distress in men with breast cancer. AB - PURPOSE: This article reports the first United Kingdom study to examine the prevalence of psychological distress in men with breast cancer and the factors associated with increased distress. PATIENTS AND METHODS: One hundred and sixty one men with breast cancer completed a cross-sectional questionnaire that included measures of anxiety and depressive symptoms, cancer-specific distress, body image, coping, information and support needs, and clinical and demographic variables. RESULTS: Clinical levels of anxiety and depressive symptoms were reported by 6% and 1% of men, respectively, while 23% reported high levels of cancer-specific distress. Anxiety was most strongly associated with avoidance coping and fear and uncertainty about the future (42% of the variance in anxiety scores, P < .001). Depressive symptoms were associated with altered body image (35% of the variance, P < .001). Body image, avoidance coping, referral to the study by a clinician, fear and uncertainty, and wanting to receive more gender specific information together explained 51% of the variance in cancer-related distress (P < .001). Clinical and demographic factors did not account for a significant proportion of the variance in any of the distress measures. CONCLUSION: Although the prevalence of clinical anxiety and depressive symptoms were low in this sample, almost a quarter of men experienced traumatic stress symptoms specific to breast cancer. Potential risk factors for distress include the use of avoidant coping strategies, negative body image, feelings of fear and uncertainty in relation to breast cancer, and unmet information needs. Suggestions are made for improving the information and support available to men with breast cancer. PMID- 16382119 TI - Successful chemotherapeutic modality of doxorubicin plus dacarbazine for the treatment of desmoid tumors in association with familial adenomatous polyposis. AB - PURPOSE: Desmoid tumors are locally aggressive and can be fatal in familial adenomatous polyposis (FAP) patients if they are not suitable for surgery or radiation therapy. Here, we prospectively investigated the efficacy of a chemotherapeutic regimen involving doxorubicin (DOX) and dacarbazine (DTIC) for inoperable FAP-associated desmoid tumors. PATIENTS AND METHODS: From an initial group of 120 FAP patients, seven of the 11 individuals with symptomatic unresectable desmoid tumors that were unresponsive to conventional hormone therapy were enrolled onto this study. The general chemotherapy regimen comprised four or five cycles of DOX (20 mg/m2 daily) plus DTIC (150 mg/m2 daily) throughout 4 days of drip intravenous infusion (day 1 through 4) every 28 days, followed by the cyclooxygenase-2 inhibitor meloxicam (10 mg/m2). The primary end point was relapse-free survival. The secondary end points included toxicity, clinical improvement, and tumor regression according to computed tomography. RESULTS: Significant tumor regression was observed clinically and radiologically in all seven patients. Three patients showed a complete response. The average progression-free survival period was 74.0 months (range, 32.5 to 107.5 months). Three patients showed grade 3 adverse events with no treatment-related mortality. All seven patients survived and remained without tumor progression. An adenomatous polyposis coli germline-mutation analysis revealed no mutations in the specified regions. CONCLUSION: A chemotherapeutic regimen of DOX plus DTIC followed by meloxicam is an effective and safe treatment for FAP-associated desmoid tumors. This modality should be considered for use as first-line chemotherapy in symptomatic desmoid tumors that are unresponsive to conventional medical therapy, due to the absence of useful presymptomatic markers. PMID- 16382121 TI - Low value of [18F]-fluoro-2-deoxy-D-glucose positron emission tomography in primary staging of early-stage cervical cancer before radical hysterectomy. AB - PURPOSE: The role of positron emission tomography (PET) with [18F]-fluoro-2-deoxy D-glucose (FDG) in early-stage cervical cancer is unclear. We aimed to investigate the clinical benefit of FDG-PET in primary staging before radical hysterectomy and pelvic lymphadenectomy (RH-PLND). PATIENTS AND METHODS: Patients with untreated stage IA2 to IIA adenocarcinoma (AD) or adenosquamous carcinoma (ASC) or nonbulky (< or = 4 cm) squamous cell carcinoma cervical cancer with magnetic resonance imaging (MRI) -defined negative nodal metastasis were enrolled onto a prospective study with a two-stage design. All patients had a preoperative dual-phase FDG-PET, technetium-99m-sulfur colloid lymphoscintigraphy, and intraoperative sentinel lymph node (LN) detection at RH-PLND. The gold standard of LN metastasis is histologic. A sample size of 120 patients was calculated to fit study aims (diagnostic efficacy of PET and sentinel LN sampling). An interim analysis was performed when 60 patients were accrued, which led to the current report. RESULTS: There were 36 SCCs, 20 ADs, and four ASCs. Of the 60 patients, 10 (16.7%) had pelvic LN metastases, and one (1.7%) had para-aortic LN (PALN) metastasis histologically. FDG-PET detected the single PALN metastasis (one of one patient) but detected only one (10%) of the 10 pelvic LN metastases. The PET false-negative pelvic LN micrometastases measured a median of 4.0 x 3.0 mm (range, 0.5 x 0.5 to 7 x 6 mm). The second stage of this trial will be continued without PET. CONCLUSION: This study shows that dual-phase FDG-PET has little value in primary, nonbulky, stage IA2 to IIA and MRI-defined, LN-negative cervical cancer. PMID- 16382120 TI - Salvage radioimmunotherapy with murine iodine-131-labeled antitenascin monoclonal antibody 81C6 for patients with recurrent primary and metastatic malignant brain tumors: phase II study results. AB - PURPOSE: To assess the efficacy and toxicity of intraresection cavity iodine-131 labeled murine antitenascin monoclonal antibody 81C6 (131I-m81C6) among recurrent malignant brain tumor patients. PATIENTS AND METHODS: In this phase II trial, 100 mCi of 131I-m81C6 was injected directly into the surgically created resection cavity (SCRC) of 43 patients with recurrent malignant glioma (glioblastoma multiforme [GBM], n = 33; anaplastic astrocytoma [AA], n = 6; anaplastic oligodendroglioma [AO], n = 2; gliosarcoma [GS], n = 1; and metastatic adenocarcinoma, n = 1) followed by chemotherapy. RESULTS: With a median follow-up of 172 weeks, 63% and 59% of patients with GBM/GS and AA/AO tumors were alive at 1 year. Median overall survival for patients with GBM/GS and AA/AO tumors was 64 and 99 weeks, respectively. Ten patients (23%) developed acute hematologic toxicity. Five patients (12%) developed acute reversible neurotoxicity. One patient (2%) developed irreversible neurotoxicity. No patients required reoperation for radionecrosis. CONCLUSION: In this single-institution phase II study, administration of 100 mCi of 131I-m81C6 to recurrent malignant glioma patients followed by chemotherapy is associated with a median survival that is greater than that of historical controls treated with surgery plus iodine-125 brachytherapy. Furthermore, toxicity was acceptable. Administration of a fixed millicurie dose resulted in a wide range of absorbed radiation doses to the SCRC. We are now conducting a phase II trial, approved by the US Food and Drug Administration, using patient-specific 131I-m81C6 dosing, to deliver 44 Gy to the SCRC followed by standardized chemotherapy. A phase III multicenter trial with patient-specific dosing is planned. PMID- 16382122 TI - Preliminary results on safety and activity of a randomized, double-blind, 2 x 2 trial of low-dose tamoxifen and fenretinide for breast cancer prevention in premenopausal women. AB - PURPOSE: To determine whether low-dose tamoxifen and fenretinide have a synergistic effect on surrogate biomarkers, including circulating insulin-like growth factor I (IGF-I) and mammographic density, in premenopausal women at risk for breast cancer and to study drug safety. PATIENTS AND METHODS: Premenopausal women (n = 235) were randomly assigned in a double-blind four-arm trial to receive tamoxifen 5 mg/d, fenretinide 200 mg/d, both agents, or placebo for 2 years. The present analysis refers to preliminary data on safety, IGF-I, and breast cancer events. RESULTS: Patients were included if they had an excised ductal carcinoma-in-situ (57%), lobular carcinoma-in-situ (13%), minimal invasive breast cancer (7%), or a 5-year Gail risk > or = 1.3% (23%). After a median follow-up of 40 months, there was a reduction of 13%, 2%, 20%, and 1% in IGF-I levels for patients on tamoxifen, fenretinide, tamoxifen plus fenretinide, and placebo, respectively. Recruitment was stopped based on the lack of an interaction on IGF-I levels, which was a primary end point for the study. Thirty six patients have dropped out of the study, 17 because of adverse events and 19 for various other reasons. One stage I endometrial cancer occurred in a patient on fenretinide, and one optic nerve ischemia and one deep venous thrombosis occurred on tamoxifen. There was no difference in menopausal symptoms, endometrial thickness, polyps, or ovarian cysts among treatment arms. To date, 24 breast cancers have been observed, without differences among arms. CONCLUSION: The combination of low-dose tamoxifen and fenretinide is safe but not synergistic in lowering IGF-I levels in premenopausal women. The clinical implications require further follow-up. PMID- 16382123 TI - Unconventional anticancer agents: a systematic review of clinical trials. AB - PURPOSE: A substantial number of cancer patients turn to treatments other than those recommended by mainstream oncologists in an effort to sustain tumor remission or halt the spread of cancer. These unconventional approaches include botanicals, high-dose nutritional supplementation, off-label pharmaceuticals, and animal products. The objective of this study was to review systematically the methodologies applied in clinical trials of unconventional treatments specifically for cancer. METHODS: MEDLINE 1966 to 2005 was searched using approximately 200 different medical subject heading terms (eg, alternative medicine) and free text words (eg, laetrile). We sought prospective clinical trials of unconventional treatments in cancer patients, excluding studies with only symptom control or nonclinical (eg, immune) end points. Trial data were extracted by two reviewers using a standardized protocol. RESULTS: We identified 14,735 articles, of which 214, describing 198 different clinical trials, were included. Twenty trials were phase I, three were phase I and II, 70 were phase II, and 105 were phase III. Approximately half of the trials investigated fungal products, 20% investigated other botanicals, 10% investigated vitamins and supplements, and 10% investigated off-label pharmaceuticals. Only eight of the phase I trials were dose-finding trials, and a mere 20% of phase II trials reported a statistical design. Of the 27 different agents tested in phase III, only one agent had a prior dose-finding trial, and only for three agents was the definitive study initiated after the publication of phase II data. CONCLUSION: Unconventional cancer treatments have not been subject to appropriate early-phase trial development. Future research on unconventional therapies should involve dose-finding and phase II studies to determine the suitability of definitive trials. PMID- 16382124 TI - Comparable long-term survival after unrelated and HLA-matched sibling donor hematopoietic stem cell transplantations for acute leukemia in children younger than 18 months. AB - PURPOSE: To describe outcomes after unrelated donor stem cell transplantation (HCT) in children (< 18 months at diagnosis) with acute leukemia and compare these with outcomes after human leukocyte antigen (HLA)-matched sibling donor HCT. PATIENTS AND METHODS: We compared the results of unrelated donor HCT with bone marrow (n = 85) or cord blood grafts (n = 81) and HLA-matched sibling donor HCT with bone marrow grafts (n = 101) for acute myeloid or acute lymphoblastic leukemia using Cox proportional hazards models. Unrelated donor HCT recipients were younger, more likely to have MLL gene rearrangement, to have advanced leukemia, and to receive irradiation before HCT. RESULTS: Treatment-related mortality rates were 6%, 15%, and 31% after matched sibling, unrelated donor bone marrow, and cord blood HCT, respectively. Risks of relapse, overall and leukemia free survival were significantly associated with disease status at transplantation. Though leukemia recurrence was lowest after unrelated donor HCT in first clinical remission (CR), overall survival, and leukemia-free survival rates were similar after matched sibling and unrelated donor HCT, after adjustment for disease status. Relapse, overall and leukemia-free survival did not differ by graft type (bone marrow v cord blood) or type of leukemia. Three year probabilities of leukemia-free survival were 49% and 54% after HLA-matched sibling and unrelated donor transplantation in first CR, respectively. Corresponding rates for those with advanced leukemia were 20% and 30%. CONCLUSION: Unrelated donor HCT should be considered for infants with acute leukemia in first CR using the same eligibility criteria as are currently used for those with HLA matched sibling donors. PMID- 16382125 TI - Intensive therapy with growth factor support for patients with Ewing tumor metastatic at diagnosis: Pediatric Oncology Group/Children's Cancer Group Phase II Study 9457--a report from the Children's Oncology Group. AB - PURPOSE: Prognosis is poor for Ewing sarcoma patients with metastasis at diagnosis. We intensified a five-drug therapy (ifosfamide, etoposide alternated with vincristine, doxorubicin, and cyclophosphamide) using filgrastim but not stem-cell support. We studied topotecan alone and combined with cyclophosphamide in therapeutic windows before the five-drug therapy. A randomly assigned proportion of patients received amifostine as a cytoprotective agent. PATIENTS AND METHODS: Eligible patients were < or = 30 years old and had histologically proven Ewing sarcoma or primitive neuroectodermal tumor (PNET) and metastasis at diagnosis. Chemotherapeutic cycles began every 21 days, after recovery from toxicities. RESULTS: One hundred ten of the 117 patients enrolled were eligible. Thirty-six patients received initial topotecan. Three had partial responses (PRs), and 17 had progressive disease (PD). Thirty-seven patients were administered topotecan and cyclophosphamide; 21 of these patients achieved PR, and one patient had PD. In a randomly assigned group of 69 patients, amifostine did not provide myeloprotection, which was measured by absolute neutrophil count, platelet count, or cycle intervals. The best responses to the overall therapy included 45 complete responses, 41 PRs, stable disease in 14 patients, and PD in five patients. For all patients, the 2-year event-free survival (EFS) rate was 24% (+/- 4%), and the overall survival rate was 46% (+/- 5%). For the 39 patients with isolated pulmonary metastases, the 2-year EFS rate was 31% (+/- 7%) compared with 20% (+/- 5%) for patients with more widespread disease. CONCLUSION: Topotecan had limited activity in patients with Ewing sarcoma or PNET metastatic at diagnosis. The topotecan-cyclophosphamide combination was active. Amifostine was not myeloprotective. Overall results showed no improvement compared with previous studies. PMID- 16382126 TI - Decision making in pediatric oncology: who should take the lead? The decisional priority in pediatric oncology model. AB - Decision making in pediatric oncology can look different to the ethicist and the clinician. Popular ethical theories argue that clinicians should not make decisions for patients, but rather provide information so that patients can make their own decisions. However, this theory does not always reflect clinical reality. We present a new model of decision making that reconciles this apparent discrepancy. We first distinguish decisional priority from decisional authority. The person (parent, child, or clinician) who first identifies a preferred choice exercises decisional priority. In contrast, decisional authority is a nondelegable parental right and duty, in which a mature child may join. This distinction enables us to analyze decisional priority without diminishing parental authority. This model analyzes decisions according to two continuous underlying characteristics. One dominant characteristic is the likelihood of cure. Because cure, when possible, is the ultimate goal, the clinician is in a better position to assume decisional priority when a child probably can be cured. The second characteristic is whether there is more than one reasonable treatment option. The interaction of these two complex continual results in distinctive types of decisional situations. This model explains why clinicians sometimes justifiably assume decisional priority when there is one best medical choice. It also suggests that clinicians should particularly encourage parents (and children, when appropriate) to assume decisional priority when there are two or more clinically reasonable choices. In this circumstance, the family, with its deeper understanding of the child's nature and preferences, is better positioned to take the lead. PMID- 16382127 TI - Radiotherapy as primary treatment for stage IE and IIE nasal natural killer/T cell lymphoma. AB - PURPOSE The optimal therapy remains unclear for nasal natural killer (NK)/T-cell lymphoma. The purpose of this study is to analyze the outcome of radiotherapy as the primary treatment for localized stage IE and IIE diseases. PATIENTS AND METHODS: One hundred five patient cases were reviewed. There were 83 stage IE and 22 stage IIE patients. All except three patients received radiotherapy (RT) alone or RT combined with chemotherapy (CT; combined-modality therapy [CMT]). Overall, 31 patients were treated with RT alone, 34 with RT followed by CT, 37 with CT followed by RT, and three with CT alone. RESULTS: Five-year overall survival (OS) and progression-free survival (PFS) for all patients were 71% and 59%, respectively. The 5-year OS and PFS were 78% and 63% for stage IE, and 46% and 40% for stage IIE, respectively. Complete response (CR) was achieved in 91 patients (87%) after RT and/or CT. Initial RT resulted in a superior CR as compared with initial CT, with 54 (83%) of 65 patients achieving CR with initial RT, versus only eight (20%) of 40 after initial CT. For 102 patients who received RT with or without CT, the outcome of primary treatment with RT alone was compared with that of CMT. Five-year OS and PFS was 66% and 61% for RT alone, and 76% and 61%% for CMT, respectively (OS, P = .6433; PFS, P = .8391). CONCLUSION: RT as primary therapy resulted in good outcome in early-stage disease, and the addition of CT to RT was not accompanied by an improvement in survival. PMID- 16382128 TI - The best guess approach to phase I trial design. PMID- 16382129 TI - Prognostic factors in human herpesvirus 8-related lymphoproliferative disorders associated with HIV infection. PMID- 16382130 TI - International Breast Cancer Study Group Trial of sentinel node biopsy. PMID- 16382131 TI - Prevention of pertussis among adolescents: recommendations for use of tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis (Tdap) vaccine. AB - The purpose of this statement is to provide the rationale and recommendations for adolescent use of tetanus toxoid, reduced diphtheria toxoid, and acellular pertussis (Tdap) vaccines. Despite universal immunization of children with multiple doses of pediatric diphtheria and tetanus toxoids and acellular pertussis (DTaP) vaccine, pertussis remains endemic with a steady increase in the number of reported cases. Two peaks in the incidence of pertussis occur in pediatric patients: infants younger than 6 months of age who are inadequately protected by the current immunization schedule and adolescents 11 through 18 years of age whose vaccine-induced immunity has waned. Significant medical and public health resources are being consumed in postexposure management of adolescent cases, contacts, and outbreaks with little beneficial effect on individuals or the epidemiology of disease. Two Tdap products were licensed in 2005 for use in people 10 through 18 years of age (Boostrix) and 11 through 64 years of age (Adacel). The American Academy of Pediatrics recommends the following: 1. Adolescents 11 to 18 years of age should receive a single dose of Tdap instead of tetanus and diphtheria toxoids (Td) vaccine for booster immunization. The preferred age for Tdap immunization is 11 to 12 years. 2. Adolescents 11 to 18 years of age who have received Td but not Tdap are encouraged to receive a single dose of Tdap. An interval of at least 5 years between Td and Tdap is suggested to reduce the risk of local and systemic reactions; however, intervals of less than 5 years can be used, particularly in settings of increased risk of acquiring pertussis, having complicated disease, or transmitting infection to vulnerable contacts. Data support acceptable safety with an interval as short as approximately 2 years. 3. Tdap and tetravalent meningococcal conjugate vaccine (MCV4 [Menactra]) should be administered during the same visit if both vaccines are indicated. If this is not feasible, MCV4 and Tdap can be administered using either sequence. When not administered simultaneously, the American Academy of Pediatrics suggests a minimum interval of 1 month between vaccines. The rationale for this strategy is to provide direct protection of immunized adolescents. With implementation of vaccine recommendations, indirect benefitalso is likely to extend to unimmunized peers and other age groups. The strategy of universal Tdap immunization at 11 to 12 years of age is cost-effective. PMID- 16382132 TI - Activation of the epidermal growth factor (EGF) receptor induces formation of EGF receptor- and Grb2-containing clathrin-coated pits. AB - In HeLa cells depleted of adaptor protein 2 complex (AP2) by small interfering RNA (siRNA) to the mu2 or alpha subunit or by transient overexpression of an AP2 sequestering mutant of Eps15, endocytosis of the transferrin receptor (TfR) was strongly inhibited. However, epidermal growth factor (EGF)-induced endocytosis of the EGF receptor (EGFR) was inhibited only in cells where the alpha subunit had been knocked down. By immunoelectron microscopy, we found that in AP2-depleted cells, the number of clathrin-coated pits was strongly reduced. When such cells were incubated with EGF, new coated pits were formed. These contained EGF, EGFR, clathrin, and Grb2 but not the TfR. The induced coated pits contained the alpha subunit, but labeling density was reduced compared to control cells. Induction of clathrin-coated pits required EGFR kinase activity. Overexpression of Grb2 with inactivating point mutations in N- or C-terminal SH3 domains or in both SH3 domains inhibited EGF-induced formation of coated pits efficiently, even though Grb2 SH3 mutations did not block activation of mitogen-activated protein kinase (MAPK) or phosphatidylinositol 3-kinase (PI3K). Our data demonstrate that EGFR induced signaling and Grb2 are essential for formation of clathrin-coated pits accommodating the EGFR, while activation of MAPK and PI3K is not required. PMID- 16382133 TI - The transcriptional histone acetyltransferase cofactor TRRAP associates with the MRN repair complex and plays a role in DNA double-strand break repair. AB - Transactivation-transformation domain-associated protein (TRRAP) is a component of several multiprotein histone acetyltransferase (HAT) complexes implicated in transcriptional regulation. TRRAP was shown to be required for the mitotic checkpoint and normal cell cycle progression. MRE11, RAD50, and NBS1 (product of the Nijmegan breakage syndrome gene) form the MRN complex that is involved in the detection, signaling, and repair of DNA double-strand breaks (DSBs). By using double immunopurification, mass spectrometry, and gel filtration, we describe the stable association of TRRAP with the MRN complex. The TRRAP-MRN complex is not associated with any detectable HAT activity, while the isolated other TRRAP complexes, containing either GCN5 or TIP60, are. TRRAP-depleted extracts show a reduced nonhomologous DNA end-joining activity in vitro. Importantly, small interfering RNA knockdown of TRRAP in HeLa cells or TRRAP knockout in mouse embryonic stem cells inhibit the DSB end-joining efficiency and the precise nonhomologous end-joining process, further suggesting a functional involvement of TRRAP in the DSB repair processes. Thus, TRRAP may function as a molecular link between DSB signaling, repair, and chromatin remodeling. PMID- 16382134 TI - RACK1 recruits STAT3 specifically to insulin and insulin-like growth factor 1 receptors for activation, which is important for regulating anchorage-independent growth. AB - Current understanding of the activation of STATs is through binding between the SH2 domain of STATs and phosphotyrosine of tyrosine kinases. Here we demonstrate a novel role of RACK1 as an adaptor for insulin and insulin-like growth factor 1 receptor (IGF-1R)-mediated STAT3 activation specifically. Intracellular association of RACK1 via its N-terminal WD domains 1 to 4 (WD1-4) with insulin receptor (IR)/IGF-1R is augmented upon respective ligand stimulation, whereas association with STAT3 is constitutive. Purified RACK1 or RACK1 WD1-4 associates directly with purified IR, IGF-1R, and STAT3 in vitro. Insulin induces multiprotein complex formation of RACK1, IR, and STAT3. Overexpression or downregulation of RACK1 greatly enhances or decreases, respectively, IR/IGF-1R mediated activation of STAT3 and its target gene expression. Site-specific mutants of IR and IGF-1R impaired in RACK1 binding are ineffective in mediating recruitment and activation of STAT3 as well as in insulin- or IGF-1-induced protection of cells from anoikis. RACK1-mediated STAT3 activation is important for insulin and IGF-1-induced anchorage-independent growth in certain ovarian cancer cells. We conclude that RACK1 mediates recruitment of STAT3 to IR and IGF 1R specifically for activation, suggesting a general paradigm for the need of an adaptor in mediating activation of STATs by receptor protein tyrosine kinases. PMID- 16382135 TI - Fanconi anemia proteins are required to prevent accumulation of replication associated DNA double-strand breaks. AB - Fanconi anemia (FA) is a multigene cancer susceptibility disorder characterized by cellular hypersensitivity to DNA interstrand cross-linking agents such as mitomycin C (MMC). FA proteins are suspected to function at the interface between cell cycle checkpoints, DNA repair, and DNA replication. Using replicating extracts from Xenopus eggs, we developed cell-free assays for FA proteins (xFA). Recruitment of the xFA core complex and xFANCD2 to chromatin is strictly dependent on replication initiation, even in the presence of MMC indicating specific recruitment to DNA lesions encountered by the replication machinery. The increase in xFA chromatin binding following treatment with MMC is part of a caffeine-sensitive S-phase checkpoint that is controlled by xATR. Recruitment of xFANCD2, but not xFANCA, is dependent on the xATR-xATR-interacting protein (xATRIP) complex. Immunodepletion of either xFANCA or xFANCD2 from egg extracts results in accumulation of chromosomal DNA breaks during replicative synthesis. Our results suggest coordinated chromatin recruitment of xFA proteins in response to replication-associated DNA lesions and indicate that xFA proteins function to prevent the accumulation of DNA breaks that arise during unperturbed replication. PMID- 16382136 TI - Distinct paths to stop codon reassignment by the variant-code organisms Tetrahymena and Euplotes. AB - The reassignment of stop codons is common among many ciliate species. For example, Tetrahymena species recognize only UGA as a stop codon, while Euplotes species recognize only UAA and UAG as stop codons. Recent studies have shown that domain 1 of the translation termination factor eRF1 mediates stop codon recognition. While it is commonly assumed that changes in domain 1 of ciliate eRF1s are responsible for altered stop codon recognition, this has never been demonstrated in vivo. To carry out such an analysis, we made hybrid proteins that contained eRF1 domain 1 from either Tetrahymena thermophila or Euplotes octocarinatus fused to eRF1 domains 2 and 3 from Saccharomyces cerevisiae. We found that the Tetrahymena hybrid eRF1 efficiently terminated at all three stop codons when expressed in yeast cells, indicating that domain 1 is not the sole determinant of stop codon recognition in Tetrahymena species. In contrast, the Euplotes hybrid facilitated efficient translation termination at UAA and UAG codons but not at the UGA codon. Together, these results indicate that while domain 1 facilitates stop codon recognition, other factors can influence this process. Our findings also indicate that these two ciliate species used distinct approaches to diverge from the universal genetic code. PMID- 16382137 TI - BS69, a specific adaptor in the latent membrane protein 1-mediated c-Jun N terminal kinase pathway. AB - We previously demonstrated that the Epstein-Barr virus-encoded latent membrane protein 1 (LMP1) potently activates the cellular c-Jun N-terminal kinase (JNK) pathway by sequentially engaging an unknown adaptor, TRAF6, TAB1/TAK1, and JNKKs. We now show that BS69, a MYND domain-containing cellular protein, is the missing adaptor that bridges LMP1 and TRAF6, as the MYND domain and a separate region of BS69 bind to the carboxyl termini of LMP1 and TRAF6, respectively. While LMP1 promotes the interaction between BS69 and TRAF6, the complex formation between LMP1 and TRAF6 is BS69 dependent. A fraction of LMP1 and BS69 is constitutively colocalized in the membrane lipid rafts. Importantly, knockdown of BS69 by small interfering RNAs specifically inhibits JNK activation by LMP1 but not tumor necrosis factor alpha. Although overexpression of either BS69 or a mutant LMP1 without the cytoplasmic carboxyl tail is not sufficient to activate JNK, interestingly, when BS69 is covalently linked to the mutant LMP1, the chimeric protein restores the ability to activate JNK. This indicates that the recruitment and aggregation of BS69 is a prerequisite for JNK activation by LMP1. PMID- 16382138 TI - IkappaB kinase alpha-mediated derepression of SMRT potentiates acetylation of RelA/p65 by p300. AB - Over the last several years, significant progress has been made in identifying chromatin-regulated events that govern NF-kappaB transcription. Using either laminin attachment or tumor necrosis factor alpha as a physiological stimulus of NF-kappaB activation, we demonstrate that IkappaB kinase alpha (IKKalpha) is recruited to chromatin in distinct phases. In the initial phase, IKKalpha is responsible for derepressing the silencing mediator for retinoic acid and thyroid hormone receptor (SMRT)-histone deacetylase 3 (HDAC3) corepressor complex from the p50 homodimer. However, in the latter phase, chromatin-bound IKKalpha coordinates the simultaneous phosphorylation of RelA/p65(S536) and SMRT(S2410) as detected by chromatin immunoprecipitation (ChIP) assays. Although phosphorylated SMRT remains bound to the active p50-RelA/p65 heterodimer of NF-kappaB, derepression of SMRT is evidenced by the loss of chromatin-associated HDAC3 activity. ChIP and re-ChIP analysis demonstrates that phosphorylation of RelA/p65(S536) and SMRT(S2410) occurs prior to acetylation of RelA/p65 at K310. Moreover, IKKalpha-induced phosphorylation of RelA/p65(S536) displaces corepressor activity, allowing p300-mediated acetylation of RelA/p65. Introduction of nonphosphorylatable mutants of RelA/p65 and SMRT proteins or the inhibition of IKK activity results in active repression of NF-kappaB promoters by tethering the SMRT-HDAC3 complex. Similar to phosphorylation within the Rel homology domain of RelA/p65, which governs an exchange of HDAC1 for CBP/p300 acetyltransferases, we demonstrate that phosphorylation within the transactivation domain of RelA/p65(S536) displaces SMRT-HDAC3 repressor activity, allowing p300 to acetylate RelA/p65. PMID- 16382139 TI - Ube1L and protein ISGylation are not essential for alpha/beta interferon signaling. AB - The expression of ubiquitin-like modifier ISG15 and its conjugation to target proteins are highly induced by interferon (IFN) stimulation and during viral and bacterial infections. However, the biological significance of this modification has not been clearly understood. To investigate the function of protein modification by ISG15, we generated a mouse model deficient in UBE1L, an ISG15 activating enzyme. Ube1L-/- mice did not produce ISG15 conjugates but expressed free ISG15 normally. ISGylation has been implicated in the reproduction and innate immunity. However, Ube1L-/- mice were fertile and exhibited normal antiviral responses against vesicular stomatitis virus and lymphocytic choriomeningitis virus infection. Our results indicate that UBE1L and protein ISGylation are not critical for IFN-alpha/beta signaling via JAK/STAT activation. Moreover, using Ube1L/Ubp43 double-deficient mice, we showed that lack of UBP43, but not the increase of protein ISGylation, is related to the increased IFN signaling in Ubp43-deficient mice. PMID- 16382140 TI - Altered RNA editing in mice lacking ADAR2 autoregulation. AB - ADAR2 is a double-stranded-RNA-specific adenosine deaminase involved in the editing of mammalian RNAs by the site-selective conversion of adenosine to inosine. Previous studies from our laboratory have demonstrated that ADAR2 can modify its own pre-mRNA to create a proximal 3' splice site containing a noncanonical adenosine-inosine dinucleotide. Alternative splicing to this proximal acceptor adds 47 nucleotides to the mature ADAR2 transcript, thereby resulting in the loss of functional ADAR2 protein expression due to premature translation termination in an alternate reading frame. To examine whether the editing of ADAR2 transcripts represents a negative autoregulatory strategy to modulate ADAR2 protein expression, we have generated genetically modified mice in which the ability of ADAR2 to edit its own pre-mRNA has been selectively ablated by deletion of a critical sequence (editing site complementary sequence [ECS]) required for adenosine-to-inosine conversion. Here we demonstrate that ADAR2 autoediting and subsequent alternative splicing are abolished in homozygous deltaECS mice and that ADAR2 protein expression is increased in numerous tissues compared to wild-type animals. The observed increases in ADAR2 protein expression correlate with the extent of ADAR2 autoediting observed with wild-type tissues and correspond to increases in the editing of ADAR2 substrates, indicating that ADAR2 autoediting is a key regulator of ADAR2 protein expression and activity in vivo. PMID- 16382141 TI - H2A.Z functions to regulate progression through the cell cycle. AB - Histone H2A variants are highly conserved proteins found ubiquitously in nature and thought to perform specialized functions in the cell. Studies in yeast on the histone H2A variant H2A.Z have shown a role for this protein in transcription as well as chromosome segregation. Our studies have focused on understanding the role of H2A.Z during cell cycle progression. We found that htz1delta cells were delayed in DNA replication and progression through the cell cycle. Furthermore, cells lacking H2A.Z required the S-phase checkpoint pathway for survival. We also found that H2A.Z localized to the promoters of cyclin genes, and cells lacking H2A.Z were delayed in the induction of these cyclin genes. Several different models are proposed to explain these observations. PMID- 16382142 TI - Hzf, a p53-responsive gene, regulates maintenance of the G2 phase checkpoint induced by DNA damage. AB - The hematopoietic zinc finger protein, Hzf, is induced in response to genotoxic and oncogenic stress. The Hzf protein is encoded by a p53-responsive gene, and its overexpression, either in cells retaining or lacking functional 53, halts their proliferation. Enforced expression of Hzf led to the appearance of tetraploid cells with supernumerary centrosomes and, ultimately, to cell death. Eliminating Hzf mRNA expression by use of short hairpin (sh) RNAs had no overt effect on unstressed cells but inhibited the maintenance of G2 phase arrest following ionizing radiation (IR), thereby sensitizing cells to DNA damage. Canonical p53-responsive gene products such as p21Cip1 and Mdm2 were induced by IR in cells treated with Hzf shRNA. However, the reduction in the level of Hzf protein was accompanied by increased polyubiquitination and turnover of p21Cip1, an inhibitor of cyclin-dependent kinases whose expression contributes to maintaining the duration of the G2 checkpoint in cells that have sustained DNA damage. Thus, two p53-inducible gene products, Hzf and p21Cip1, act concomitantly to enforce the G(2) checkpoint. PMID- 16382143 TI - The splicing factor Prp43p, a DEAH box ATPase, functions in ribosome biogenesis. AB - Biogenesis of the small and large ribosomal subunits requires modification, processing, and folding of pre-rRNA to yield mature rRNA. Here, we report that efficient biogenesis of both small- and large-subunit rRNAs requires the DEAH box ATPase Prp43p, a pre-mRNA splicing factor. By steady-state analysis, a cold sensitive prp43 mutant accumulates 35S pre-rRNA and depletes 20S, 27S, and 7S pre rRNAs, precursors to the small- and large-subunit rRNAs. By pulse-chase analysis, the prp43 mutant is defective in the formation of 20S and 27S pre-rRNAs and in the accumulation of 18S and 25S mature rRNAs. Wild-type Prp43p immunoprecipitates pre-rRNAs and mature rRNAs, indicating a direct role in ribosome biogenesis. The Prp43p-Q423N mutant immunoprecipitates 27SA2 pre-rRNA threefold more efficiently than the wild type, suggesting a critical role for Prp43p at the earliest stages of large-subunit biogenesis. Consistent with an early role for Prp43p in ribosome biogenesis, Prp43p immunoprecipitates the majority of snoRNAs; further, compared to the wild type, the prp43 mutant generally immunoprecipitates the snoRNAs more efficiently. In the prp43 mutant, the snoRNA snR64 fails to methylate residue C2337 in 27S pre-rRNA, suggesting a role in snoRNA function. We propose that Prp43p promotes recycling of snoRNAs and biogenesis factors during pre-rRNA processing, similar to its recycling role in pre-mRNA splicing. The dual function for Prp43p in the cell raises the possibility that ribosome biogenesis and pre mRNA splicing may be coordinately regulated. PMID- 16382144 TI - Prp43p is a DEAH-box spliceosome disassembly factor essential for ribosome biogenesis. AB - The known function of the DEXH/D-box protein Prp43p is the removal of the U2, U5, and U6 snRNPs from the postsplicing lariat-intron ribonucleoprotein complex. We demonstrate that affinity-purified Prp43p-associated material includes the expected spliceosomal components; however, we also identify several preribosomal complexes that are specifically purified with Prp43p. Conditional prp43 mutant alleles confer a 35S pre-rRNA processing defect, with subsequent depletion of 27S and 20S precursors. Upon a shift to a nonpermissive temperature, both large and small-ribosomal-subunit proteins accumulate in the nucleolus of prp43 mutants. Pulse-chase analysis demonstrates delayed kinetics of 35S, 27S, and 20S pre-rRNA processing with turnover of these intermediates. Microarray analysis of pre-mRNA splicing defects in prp43 mutants shows a very mild effect, similar to that of nonessential pre-mRNA splicing factors. Prp43p is the first DEXH/D-box protein shown to function in both RNA polymerase I and polymerase II transcript metabolism. Its essential function is in its newly characterized role in ribosome biogenesis of both ribosomal subunits, positioning Prp43p to regulate both pre mRNA splicing and ribosome biogenesis. PMID- 16382146 TI - Elimination of epiplakin by gene targeting results in acceleration of keratinocyte migration in mice. AB - Epiplakin (EPPK) was originally identified as a human epidermal autoantigen. To identify the function of epiplakin, we generated epiplakin "knockout" mice. These mice developed normally, with apparently normal epidermis and hair. Electron microscopy after immunostaining revealed the presence of EPPK adjacent to keratin filaments in wild-type mice, suggesting that epiplakin might associate with keratin. The appearance and localization of keratin bundles in intact epidermal keratinocytes of EPPK-/- mice were similar to those in wild-type mice. Wounds on the backs of EPPK-/- mice closed more rapidly than those on the backs of wild type and heterozygous mice. The outgrowth of keratinocytes from skin explants from knockout mice was enhanced compared to outgrowth from explants from wild type mice, even in the presence of mitomycin C, suggesting that the difference in keratinocyte outgrowth might be due to a difference in the speed of migration of keratinocytes. At wound edges in wild-type mice, EPPK was expressed in proliferating keratinocytes in conjunction with keratin 6. In EPPK-/- mice, no similar proliferating keratinocytes were observed, but migrating keratinocytes weakly expressed keratin 6. EPPK was coexpressed with keratin 6 in some keratinocytes in explant cultures from wild mice. We propose that EPPK might be linked functionally with keratin 6. PMID- 16382145 TI - Regulation of sexual dimorphism: mutational and chemogenetic analysis of the doublesex DM domain. AB - Doublesex (dsx) is a transcription factor in Drosophila that regulates somatic sexual differentiation. Male- and female-specific splicing isoforms of DSX share a novel DNA-binding domain, designated the DM motif. Broadly conserved among metazoan sex-determining factors, the DM domain contains a nonclassical zinc module and binds in the DNA minor groove. Here, we characterize the DM motif by site-directed and random mutagenesis using a yeast one-hybrid (Y1H) system and extend this analysis by chemogenetic complementation in vitro. The Y1H system is based on a sex-specific Drosophila enhancer element and validated through studies of intersexual dsx mutations. We demonstrate that the eight motif-specific histidines and cysteines engaged in zinc coordination are each critical and cannot be interchanged; folding also requires conserved aliphatic side chains in the hydrophobic core. Mutations that impair DNA binding tend to occur at conserved positions, whereas neutral substitutions occur at nonconserved sites. Evidence for a specific salt bridge between a conserved lysine and the DNA backbone is obtained through the synthesis of nonstandard protein and DNA analogs. Together, these results provide molecular links between the structure of the DM domain and its function in the regulation of sexual dimorphism. PMID- 16382147 TI - Epiplakin is dispensable for skin barrier function and for integrity of keratin network cytoarchitecture in simple and stratified epithelia. AB - Epiplakin, a giant epithelial protein of >700 kDa, belongs to the plakin family of cytolinker proteins. It represents an atypical family member, however, as it consists entirely of plakin repeat domains but lacks any of the other domains commonly shared by plakins. Hence, its putative function as a cytolinker protein remains to be shown. To investigate epiplakin's biological role, we generated epiplakin-deficient mice by gene targeting in embryonic stem cells. Epiplakin deficient mice were viable and fertile, without developing any discernible phenotype. Ultrastructurally, their epidermis revealed no differences compared to wild-type littermates, and cornified envelopes isolated from skin showed no alterations in shape or stability. Furthermore, neither embryonal formation nor later function of the epithelial barrier was affected. In primary cultures of epiplakin-deficient keratinocytes, the organization of actin filaments, microtubules, and keratin networks was found to be normal. Similarly, no alterations in keratin network organization were observed in simple epithelia of small intestine and liver or in primary hepatocytes. We conclude that, despite epiplakin's abundant and highly specific expression in stratified and simple epithelia, its absence in mice does not lead to severe skin dysfunctions, nor has it detectable consequences for keratin filament organization and cytoarchitecture of cells. PMID- 16382148 TI - Tissue transglutaminase serves as an inhibitor of apoptosis by cross-linking caspase 3 in thapsigargin-treated cells. AB - Thapsigargin (THG) is an inhibitor of the endoplasmic reticulum Ca2+-ATPase that induces caspase 3 activation and apoptosis in HCT116 cells through a Bax dependent pathway. In Bax-deficient HCT116 cells, however, THG specifically generates two additional species of caspase 3, termed p40 and p64, with molecular masses of approximately 40 and 64 kDa, respectively, through unknown mechanisms. Here, we report that the Ca2+-dependent protein cross-linking enzyme tissue transglutaminase (tTGase) is involved in THG-induced p40 and p64 formation by catalyzing caspase 3 cross-linking reactions, thereby inactivating caspase 3 and apoptosis in Bax-deficient cells. Overexpression of tTGase increases p40 and p64 in THG-treated cells, and purified tTGase catalyzes procaspase 3 cross-linking in vitro. Inhibition of tTGase activity by either the tTGase inhibitor monodansylcadaverine or short-hairpin RNA reduces the cross-linked species p40 and p64 and restores caspase 3 activation in response to THG treatment. Moreover, prolonged exposure to THG results in a decrease in protein levels of XIAP and cIAP-1, which is subsequently followed by an increase in tTGase protein expression and activity. Expression of cytosolic Smac sensitizes Bax-deficient cells to THG-induced apoptosis; however, this effect is diminished by coexpression of tTGase. Taken together, these results suggest a novel role for tTGase as a new type of caspase 3 inhibitor in THG-mediated apoptosis. PMID- 16382149 TI - Heat shock-independent induction of multidrug resistance by heat shock factor 1. AB - The screening of two different retroviral cDNA expression libraries to select genes that confer constitutive doxorubicin resistance has in both cases resulted in the isolation of the heat shock factor 1 (HSF1) transcription factor. We show that HSF1 induces a multidrug resistance phenotype that occurs in the absence of heat shock or cellular stress and is mediated at least in part through the constitutive activation of the multidrug resistance gene 1 (MDR-1). This drug resistance phenotype does not correlate with an increased expression of heat shock-responsive genes (heat shock protein genes, or HSPs). In addition, HSF1 mutants lacking HSP gene activation are also capable of conferring multidrug resistance, and only hypophosphorylated HSF1 complexes accumulate in transduced cells. Our results indicate that HSF1 can activate MDR-1 expression in a stress independent manner that differs from the canonical heat shock-activated mechanism involved in HSP induction. We further provide evidence that the induction of MDR 1 expression occurs at a posttranscriptional level, revealing a novel undocumented role for hypophosphorylated HSF1 in posttranscriptional gene regulation. PMID- 16382150 TI - Down-regulation of nucleosomal binding protein HMGN1 expression during embryogenesis modulates Sox9 expression in chondrocytes. AB - We find that during embryogenesis the expression of HMGN1, a nuclear protein that binds to nucleosomes and reduces the compaction of the chromatin fiber, is progressively down-regulated throughout the entire embryo, except in committed but continuously renewing cell types, such as the basal layer of the epithelium. In the developing limb bud, the expression of HMGN1 is complementary to Sox9, a master regulator of the chondrocyte lineage. In limb bud micromass cultures, which faithfully mimic in vivo chondrogenic differentiation, loss of HMGN1 accelerates differentiation. Expression of wild-type HMGN1, but not of a mutant HMGN1 that does not bind to chromatin, in Hmgn1-/- micromass cultures inhibits Sox9 expression and retards differentiation. Chromatin immunoprecipitation analysis reveals that HMGN1 binds to Sox9 chromatin in cells that are poised to express Sox9. Loss of HMGN1 elevates the amount of HMGN2 bound to Sox9, suggesting functional redundancy among these proteins. These findings suggest a role for HMGN1 in chromatin remodeling during embryogenesis and in the activation of Sox9 during chondrogenesis. PMID- 16382151 TI - Mutant Lrp1 knock-in mice generated by recombinase-mediated cassette exchange reveal differential importance of the NPXY motifs in the intracellular domain of LRP1 for normal fetal development. AB - Lrp1 knock-in mice carrying either a wild-type allele or three different mutated alleles encoding the multifunctional endocytic receptor LRP1 were generated by recombinase-mediated cassette exchange (RMCE). Reinsertion by RMCE of a wild-type allele led to a normal pattern and level of gene expression and a completely normal phenotype, indicating that the RMCE procedure itself is neutral with respect to the function of the gene locus. In contrast, reinsertion of mutated LRP1 alleles carrying either inactivating mutations in the proximal NPXY motif (NPTY-->AATA) of the cytoplasmic domain or in the furin cleavage site (RHRR- >AHAA) caused distinctive liver phenotypes: respectively, either a late fetal destruction of the organ causing perinatal death or a selective enlargement of von-Kupffer cell lysosomes reminiscent of a mild lysosomal storage without an apparent negative effect on animal survival. Notably, mutation of the distal NPXY motif overlapping with an YXXL motif (NPVYATL-->AAVAATL) did not cause any obvious pathological effect. The mutations showed no effect on the LRP1 expression level; however, as expected, the proteolytic maturation of LRP1 into its two subunits was significantly impaired, although not completely abolished, in the furin cleavage mutant. These data demonstrate that RMCE is a reliable and efficient approach to generate multiple mutant knock-in alleles for in vivo functional analysis of individual domains or motifs of large multidomain proteins. Its application in Lrp1 reveals dramatically variant phenotypes, of which further characterization will definitively contribute to our understanding of the biology of this multifunctional receptor. PMID- 16382152 TI - Modulation of prion formation, aggregation, and toxicity by the actin cytoskeleton in yeast. AB - Self-perpetuating protein aggregates transmit prion diseases in mammals and heritable traits in yeast. De novo prion formation can be induced by transient overproduction of the corresponding prion-forming protein or its prion domain. Here, we demonstrate that the yeast prion protein Sup35 interacts with various proteins of the actin cortical cytoskeleton that are involved in endocytosis. Sup35-derived aggregates, generated in the process of prion induction, are associated with the components of the endocytic/vacuolar pathway. Mutational alterations of the cortical actin cytoskeleton decrease aggregation of overproduced Sup35 and de novo prion induction and increase prion-related toxicity in yeast. Deletion of the gene coding for the actin assembly protein Sla2 is lethal in cells containing the prion isoforms of both Sup35 and Rnq1 proteins simultaneously. Our data are consistent with a model in which cytoskeletal structures provide a scaffold for generation of large aggregates, resembling mammalian aggresomes. These aggregates promote prion formation. Moreover, it appears that the actin cytoskeleton also plays a certain role in counteracting the toxicity of the overproduced potentially aggregating proteins. PMID- 16382153 TI - Regulation of polymerase II transcription by 7SK snRNA: two distinct RNA elements direct P-TEFb and HEXIM1 binding. AB - The positive transcription elongation factor b (P-TEFb), a complex of Cdk9 and cyclin T1/T2, stimulates transcription by phosphorylating RNA polymerase II. The 7SK small nuclear RNA, in cooperation with HEXIM1 protein, functions as a general polymerase II transcription regulator by sequestering P-TEFb into a large kinase inactive 7SK/HEXIM1/P-TEFb complex. Here, determination and characterization of the functionally essential elements of human 7SK snRNA directing HEXIM1 and P TEFb binding led to a new model for the assembly of the 7SK/HEXIM1/P-TEFb regulatory complex. We demonstrate that two structurally and functionally distinct protein binding elements located in the 5'- and 3'-terminal hairpins of 7SK support the in vivo recruitment of HEXIM1 and P-TEFb. Consistently, a minimal regulatory RNA composed of the 5' and 3' hairpins of 7SK can modulate polymerase II transcription in HeLa cells. HEXIM1 binds independently and specifically to the G24-C48/G60-C87 distal segment of the 5' hairpin of 7SK. Binding of HEXIM1 is a prerequisite for association of P-TEFb with the G302-C324 apical region of the 3' hairpin of 7SK that is highly reminiscent of the human immunodeficiency virus transactivation-responsive RNA. PMID- 16382154 TI - Rap1A-deficient T and B cells show impaired integrin-mediated cell adhesion. AB - Studies in tissue culture cells have demonstrated a role for the Ras-like GTPase Rap1 in the regulation of integrin-mediated cell-matrix and cadherin-mediated cell-cell contacts. To analyze the function of Rap1 in vivo, we have disrupted the Rap1A gene by homologous recombination. Mice homozygous for the deletion allele are viable and fertile. However, primary hematopoietic cells isolated from spleen or thymus have a diminished adhesive capacity on ICAM and fibronectin substrates. In addition, polarization of T cells from Rap1-/- cells after CD3 stimulation was impaired compared to that of wild-type cells. Despite this, these defects did not result in hematopoietic or cell homing abnormalities. Although it is possible that the relatively mild phenotype is a consequence of functional complementation by the Rap1B gene, our genetic studies confirm a role for Rap1A in the regulation of integrins. PMID- 16382155 TI - Deletion of Smad2 in mouse liver reveals novel functions in hepatocyte growth and differentiation. AB - Smad family proteins Smad2 and Smad3 are activated by transforming growth factor beta (TGF-beta)/activin/nodal receptors and mediate transcriptional regulation. Although differential functional roles of Smad2 and Smad3 are apparent in mammalian development, the relative functional roles of Smad2 and Smad3 in postnatal systems remain unclear. We used Cre/loxP-mediated gene targeting for hepatocyte-specific deletion of Smad2 (S2HeKO) in adult mice and generated hepatocyte-selective Smad2/Smad3 double knockouts by intercrossing AlbCre/Smad2(f/f) (S2HeKO) and Smad3-deficient Smad3ex8/ex8 (S3KO) mice. All strains were viable and had normal adult liver. However, necrogenic CCL4-induced hepatocyte proliferation was significantly increased in S2HeKO compared to Ctrl and S3KO livers, and transplanted S2HeKO hepatocytes repopulated recipient liver at dramatically increased rates compared to Ctrl hepatocytes in vivo. Using primary hepatocytes, we found that TGF-beta-induced G1 arrest, apoptosis, and epithelial-to-mesenchymal transition in Ctrl and S2HeKO but not in S3KO hepatocytes. Interestingly, S2HeKO cells spontaneously acquired mesenchymal features characteristic of epithelial-to-mesenchymal transition (EMT). Collectively, these results demonstrate that Smad2 suppresses hepatocyte growth and dedifferentiation independent of TGF-beta signaling. Smad2 is not required for TGF-beta-stimulated apoptosis, EMT, and growth inhibition in hepatocytes. PMID- 16382156 TI - Normal immune development and glucocorticoid-induced thymocyte apoptosis in mice deficient for the T-cell death-associated gene 8 receptor. AB - T-cell death-associated gene 8 (TDAG8) is a G-protein-coupled receptor transcriptionally upregulated by glucocorticoids (GCs) and implicated by overexpression studies in psychosine-mediated inhibition of cytokinesis and in GC induced apoptosis. To examine the physiological function of TDAG8, we generated knockout (KO) mice by homologous recombination. An enhanced green fluorescent protein reporter was knocked into the disrupted tdag8 locus to allow the analysis of TDAG8 expression in living cells. Interestingly, we found that during thymocyte development, TDAG8 expression resembled the dynamic regulation described for known modulators of GC-induced apoptosis, including Bcl-2, Notch1, and GC receptor. TDAG8 was expressed in double-negative cells, was downregulated at the double-positive transition, and was upregulated in single-positive thymocytes. However, despite this striking expression pattern, maturation and selection of thymocytes, as well as major immune functions, were not affected in TDAG8 KO mice. In contrast to previous overexpression results, TDAG8 was dispensable for psychosine-induced formation of multinucleated cells. Furthermore, TDAG8 KO thymocytes showed normal apoptosis following in vivo and in vitro GC treatment. These results, while establishing a useful reporter strain to study T-lymphocyte maturation, argue against a critical role for TDAG8 in immune development, psychosine-mediated inhibition of cytokinesis, and GC-induced cell death. PMID- 16382157 TI - Rec2 interplay with both Brh2 and Rad51 balances recombinational repair in Ustilago maydis. AB - Rec2 is the single Rad51 paralog in Ustilago maydis. Here, we find that Rec2 is required for radiation-induced Rad51 nuclear focus formation but that Rec2 foci form independently of Rad51 and Brh2. Brh2 foci also form in the absence of Rad51 and Rec2. By coprecipitation from cleared extracts prepared from Escherichia coli cells expressing the proteins, we found that Rec2 interacts physically not only with Rad51 and itself but also with Brh2. Transgenic expression of Brh2 in rec2 mutants can effectively restore radiation resistance, but the frequencies of spontaneous Rad51 focus formation and allelic recombination are elevated. The Dss1-independent Brh2-RPA70 fusion protein is also active in restoring radiation sensitivity of rec2 but is hyperactive to an extreme degree in allelic recombination and in suppressing the meiotic block of rec2. However, the high frequency of chromosome missegregation in meiotic products is an indicator of a corrupted process. The results demonstrate that the importance of Rec2 function is not only in stimulating recombination activity but also in ensuring that recombination is properly controlled. PMID- 16382158 TI - Characterization of SPAK and OSR1, regulatory kinases of the Na-K-2Cl cotransporter. AB - Our recent studies demonstrate that SPAK (Ste20p-related Proline Alanine-rich Kinase), in combination with WNK4 [With No lysine (K) kinase], phosphorylates and stimulates the Na-K-2Cl cotransporter (NKCC1), whereas catalytically inactive SPAK (K104R) fails to activate the cotransporter. The catalytic domain of SPAK contains an activation loop between the well-conserved DFG and APE motifs. We speculated that four threonine residues (T231, T236, T243, and T247) in the activation loop might be sites of phosphorylation and kinase activation; therefore, we mutated each residue into an alanine. In this report, we demonstrate that coexpression of SPAK (T243A) or SPAK (T247A) with WNK4 not only prevented, but robustly inhibited, cotransporter activity in NKCC1-injected Xenopus laevis oocytes. These activation loop mutations produced an effect similar to that of the SPAK (K104R) mutant. In vitro phosphorylation experiments demonstrate that both intramolecular autophosphorylation of SPAK and phosphorylation of NKCC1 are significantly stronger in the presence of Mn2+ rather than Mg2+. We also show that SPAK activity is markedly inhibited by staurosporine and K252a, partially inhibited by N-ethylmaleimide and diamide, and unaffected by arsenite. OSR1, a kinase closely related to SPAK, exhibited similar kinase properties and similar functional activation of NKCC1 when coexpressed with WNK4. PMID- 16382159 TI - Inhibitor of apoptosis protein cIAP2 is essential for lipopolysaccharide-induced macrophage survival. AB - The cellular inhibitor of apoptosis 2 (cIAP2/HIAP1) is a potent inhibitor of apoptotic death. In contrast to the other members of the IAP family, cIAP2 is transcriptionally inducible by nuclear factor-kappaB in response to multiple triggers. We demonstrate here that cIAP2-/- mice exhibit profound resistance to lipopolysaccharide (LPS)-induced sepsis, specifically because of an attenuated inflammatory response. We show that LPS potently upregulates cIAP2 in macrophages and that cIAP2-/- macrophages are highly susceptible to apoptosis in a LPS induced proinflammatory environment. Hence, cIAP2 is critical in the maintenance of a normal innate immune inflammatory response. PMID- 16382160 TI - The G-protein-coupled receptor GPR103 regulates bone formation. AB - GPR103 is a G-protein-coupled receptor with reported expression in brain, heart, kidney, adrenal gland, retina, and testis. It encodes a 455-amino-acid protein homologous to neuropeptide FF2, neuropeptide Y2, and galanin GalR1 receptors. Its natural ligand was recently identified as 26RFa, a novel human RF-amide-related peptide with orexigenic activity. To identify the function of GPR103, we generated GPR103-deficient mice. Homozygous mutant mice were viable and fertile. Their body weight was undistinguishable from that of their wild-type littermates. Histological analysis revealed that GPR103-/- mice exhibited a thinned osteochondral growth plate, a thickening of trabecular branches, and a reduction in osteoclast number, suggestive of an early arrest of osteochondral bone formation. Microcomputed tomography confirmed the reduction in trabecular bone and connective tissue densities in GPR103 knockout animals. Whole-body radiography followed by morphometric analysis revealed a kyphosis in mutant animals. Reverse transcription-PCR analysis showed that GPR103 was expressed in human skull, mouse spine, and several osteoblast cell lines. Dexamethasone, a known inhibitor of osteoblast growth and inducer of osteoblast differentiation, inhibited GPR103 expression in human osteoblast primary cultures. Altogether, these results suggest that osteopenia in GPR103-/- mice may be mediated directly by the loss of GPR103 expression in bone. PMID- 16382161 TI - Oligo-astheno-teratozoospermia in mice lacking RA175/TSLC1/SynCAM/IGSF4A, a cell adhesion molecule in the immunoglobulin superfamily. AB - RA175/TSLC1/SynCAM/IGSF4A (RA175), a member of the immunoglobulin superfamily with Ca2+-independent homophilic trans-cell adhesion activity, participates in synaptic and epithelial cell junctions. To clarify the biological function of RA175, we disrupted the mouse Igsf4a (Ra175/Tslc1/SynCam/Igsf4a Ra175) gene. Male mice lacking both alleles of Ra175 (Ra175-/-) were infertile and showed oligo astheno-teratozoospermia; almost no mature motile spermatozoa were found in the epididymis. Heterozygous males and females and homozygous null females were fertile and had no overt developmental defects. RA175 was mainly expressed on the cell junction of spermatocytes, elongating and elongated spermatids (steps 9 to 15) in wild-type testes; the RA175 expression was restricted to the distal site (tail side) but not to the proximal site (head side) in elongated spermatids. In Ra175-/- testes, elongated and mature spermatids (steps 13 to 16) were almost undetectable; round spermatids were morphologically normal, but elongating spermatids (steps 9 to 12) failed to mature further and to translocate to the adluminal surface. The remaining elongating spermatids at improper positions were finally phagocytosed by Sertoli cells. Furthermore, undifferentiated and abnormal spermatids exfoliated into the tubular lumen from adluminal surfaces. Thus, RA175 based cell junction is necessary for retaining elongating spermatids in the invagination of Sertoli cells for their maturation and translocation to the adluminal surface for timely release. PMID- 16382162 TI - RalB mobilizes the exocyst to drive cell migration. AB - The Ras family GTPases RalA and RalB have been defined as central components of the regulatory machinery supporting tumor initiation and progression. Although it is known that Ral proteins mediate oncogenic Ras signaling and physically and functionally interact with vesicle trafficking machinery, their mechanistic contribution to oncogenic transformation is unknown. Here, we have directly evaluated the relative contribution of Ral proteins and Ral effector pathways to cell motility and directional migration. Through loss-of-function analysis, we find that RalA is not limiting for cell migration in normal mammalian epithelial cells. In contrast, RalB and the Sec6/8 complex or exocyst, an immediate downstream Ral effector complex, are required for vectorial cell motility. RalB expression is required for promoting both exocyst assembly and localization to the leading edge of moving cells. We propose that RalB regulation of exocyst function is required for the coordinated delivery of secretory vesicles to the sites of dynamic plasma membrane expansion that specify directional movement. PMID- 16382163 TI - Involvement of cAMP-response element binding protein-1 in arachidonic acid induced vascular smooth muscle cell motility. AB - In addition to their role in many vital cellular functions, arachidonic acid (AA) and its eicosanoid metabolites are involved in the pathogenesis of several diseases, including atherosclerosis and cancer. To understand the potential mechanisms by which these lipid molecules could influence the disease processes, particularly cardiovascular diseases, we studied AA's effects on vascular smooth muscle cell (VSMC) motility and the role of cAMP-response element binding protein 1 (CREB-1) in this process. AA exerted differential effects on VSMC motility; at lower doses, it stimulated motility, whereas at higher doses, it was inhibitory. AA-induced VSMC motility requires its conversion via the lipoxygenase (LOX) and cyclooxygenase (COX) pathways. AA stimulated the phosphorylation of extracellular signal-regulated kinases (ERKs), Jun N-terminal kinases (JNKs), and p38 mitogen activated protein kinase (p38MAPK) in a time-dependent manner, and blockade of these serine/threonine kinases significantly attenuated AA-induced VSMC motility. In addition, AA stimulated CREB-1 phosphorylation and activity in a manner that was also dependent on its metabolic conversion via the LOX and COX pathways and the activation of ERKs and p38MAPK but not JNKs. Furthermore, suppression of CREB 1 activation inhibited AA-induced VSMC motility. 15(S)-Hydroxyeicosatetraenoic acid and prostaglandin F2alpha, the 15-LOX and COX metabolites of AA, respectively, that are produced by VSMC at lower doses, were also found to stimulate motility in these cells. Together, these results suggest that AA induces VSMC motility by complex mechanisms involving its metabolism via the LOX and COX pathways as well as the ERK- and p38MAPK-dependent and JNK-independent activation of CREB-1. PMID- 16382164 TI - A unique feeding method by a teleost fish, the fourhorn poacher Hypsagonus quadricornis (Agonidae). PMID- 16382165 TI - Hydrogen peroxide induces apoptotic-like cell death in coelomocytes of Themiste petricola (Sipuncula). AB - Apoptosis is an active form of cell death that plays a critical role in physiological and pathological conditions of multicellular organisms. These conditions include development, organogenesis, and elimination of infected, mutated, or damaged cells. Sipunculan cells may respond to changes in environmental exposure to oxidative stress by induction of apoptotic cell death. In coelomocytes of the sipunculan worm Themiste petricola, we evaluated morphological and biochemical changes that were induced by hydrogen peroxide (H2O2) and that could be compatible with an apoptotic-like phenotype. At an exposure of 100 mM H2O2, coelomocytes exhibited several morphological hallmarks of apoptosis such as chromatin condensation, nuclear segmentation, cell volume decrease, membrane blebbing, and formation of apoptotic bodies. Biochemical evidences of apoptotic-like cell death included exposure of phosphatidylserine (PS) in the outer leaflet of the plasma membrane and oligonucleosomal DNA fragmentation. In addition, exposure of coelomocytes to H2O2 induced a rapid massive loss of mitochondrial membrane potential and of the acidic pH of lysosomes. Overall, our results showed that, in sipunculan coelomocytes, H2O2 can induce changes compatible with an apoptotic-like phenotype. The finding of an oxidative-stress-induced apoptotic-like phenotype in a sipunculan worm may indicate that this kind of cell death process participates in regulation of cell number during physiological and pathological situations, including immune responses. PMID- 16382166 TI - Transdifferentiation in holothurian gut regeneration. AB - It has recently been shown that the whole spectrum of cell types constituting a multicellular organism can be generated from stem cells. Our study provides an example of an alternative mechanism of tissue repair. Injection of distilled water into the coelomic cavity of the holothurian Eupentacta fraudatrix results in the loss of the whole digestive tract, except the cloaca. The new gut reforms from two separate rudiments. One rudiment appears at the anterior end of the body and extends posteriorly. The second rudiment grows anteriorly from the cloaca. In the anterior rudiment, the luminal epithelium (normally derived from endoderm) develops de novo through direct transdifferentiation of the coelomic epithelial cells (mesodermal in origin). In the posterior rudiment, the luminal epithelium originates from the lining epithelium of the cloaca. After 27 days, the two rudiments come into contact and fuse to form a continuous digestive tube lined with a fully differentiated luminal epithelium. Thus in this species, the luminal epithelia of the anterior and posterior gut rudiments develop from two different cell sources-i.e., from the mesodermally derived mesothelium and the endodermally derived epithelium of the cloacal lining, respectively. Our data suggest that differentiated cells of echinoderms are capable of transdifferentiation into other cell types. PMID- 16382167 TI - Larval release rhythm of the mole crab Emerita talpoida (Say). AB - Ovigerous mole crabs Emerita talpoida (Say) were monitored in the laboratory to determine if the time of larval release is synchronous and under endogenous control. To determine the time of larval release, ovigerous females were placed under a 14:10 light/dark cycle simulating the ambient photoperiod. Hatching was rhythmic, occurring as a quick burst lasting about 5-15 min shortly after the onset of darkness. An individual mole crab will release batches of larvae for up to three successive nights, suggesting that the rhythm is under endogenous control. Mole crabs monitored under constant low-level red light displayed the same release pattern with hatching occurring near the time of expected sunset, indicating the presence of a circadian rhythm in larval release. To investigate whether the female or the embryos control hatching, a portion of the egg mass (50 100 embryos) was separated from the female. The time of hatching of the detached embryos subjected to either a still or shaken treatment was compared with the hatching time of embryos still attached to the female. Detached eggs in both treatments hatched within 1.5-2 h of the time of the female-attached eggs, which suggests that embryos control the timing of hatching. PMID- 16382168 TI - Spermiogenesis in the Hagfish Eptatretus burgeri (Agnatha). AB - The fine structure of spermatid differentiation in a primitive vertebrate, the hagfish, whose spermatozoa bear acrosomes, was investigated. In early round spermatids, the acrosomal vesicles were spherical and located in a shallow nuclear indentation, flanked by the plasma and the nuclear membranes. The vesicle underwent a transition through lens-shaped and cap-shaped stages until it attained the shape of a bell in mature spermatozoa. Electron-dense acrosomal material that appeared as deposits in three portions of the vesicle finally joined in the center region at a late stage. Condensation of chromatin occurred in the anterior region of the nucleus. During transformation of the spermatids, many regularly spaced microtubules appeared beneath the plasma membrane except in the anteriormost region of the cell. The microtubules in a single alignment lay parallel to one another and encased the nucleus diagonally. During an early stage, the centrioles changed their orientation from perpendicular to longitudinal and rotated to become parallel to the long axis of the nucleus. Thus, the flagellum lay nearly straight along the cell axis. A cytoplasmic canal appeared transiently during the early stage. A droplet of cytoplasm was eliminated after descending along the flagella. The features of spermiogenesis in hagfish, which lies between invertebrates and vertebrates, are compared with those of other animals. PMID- 16382169 TI - Spermatozoa and sperm aggregates in the vestimentiferan Lamellibrachia luymesi compared with those of Riftia pachyptila (Polychaeta: Siboglinidae: Vestimentifera). AB - The spermatozoa and the sperm bundles of the vestimentiferans Riftia pachyptila and Lamellibrachia luymesi (Annelida: Siboglinidae) were studied using several microscopical techniques (transmission and scanning electron microscopy, and confocal microscopy) and compared with some other annelid sperm. The spermatozoa and sperm bundles of both species show a similar structure, but they differ in the dimensions of the components of individual cells and in the number of spermatozoa forming each sperm bundle. The spermatozoa of R. pachyptila and L. luymesi are filiform cells composed, in sequence, by an acrosome in the form of a thread-like helical vesicle, an elongated coiled nucleus surrounded by two helical mitochondria, and a long flagellum. In the spermatozoa of both species, the apical portion of the nucleus is completely devoid of chromatin and is delimited by a thickened nuclear envelope with a fibrillar appearance. Both species have sperm bundles that resemble buds, having a calyx-like portion formed by the helical heads, and a stalk-like portion formed by the tightly packed flagella. A parsimony analysis based on spermatozoal characters showed monophyly of the Siboglinidae and the Vestimentifera. We propose a new set of autapomorphies characterizing vestimentiferan spermatozoa. Our analysis suggests that spermatozoal characters are useful to the understanding of the phylogeny of the group. PMID- 16382170 TI - Stable Symbiodinium composition in the sea fan Gorgonia ventalina during temperature and disease stress. AB - Like most Caribbean octocorals, Gorgonia ventalina, the common sea fan, harbors endosymbiotic dinoflagellates belonging to the genus Symbiodinium. When stressed, the host can lose these algal symbionts, a phenomenon termed "bleaching." Many cnidarians host multiple types of algal symbionts within the genus Symbiodinium, and certain types of algae may be more tolerant of stress than others. We examined the effects of temperature, temperature-induced bleaching, and infection by Aspergillus sydowii, a fungal pathogen, on Symbiodinium types harbored by the sea fan Gorgonia ventalina in the Florida Keys. Symbiont type, identified on the basis of variation in small subunit nuclear ribosomal genes or large subunit chloroplast ribosomal genes, did not vary with temperature treatment or infection status. Although allelic variation based on one microsatellite locus was found among samples and reef site, it did not consistently correlate with temperature, treatment, or disease status, suggesting that the symbiont-host relationship is stable. An aberrant PCR product was found in samples collected at one site and could be used to differentiate Symbiodinium populations among sites in the Florida Keys. PMID- 16382172 TI - Cardiovascular toxicities upon manganese exposure. AB - Manganese (Mn)-induced Parkinsonism has been well documented; however, little attention has been devoted to Mn-induced cardiovascular dysfunction. This review summarizes literature data from both animal and human studies on Mn's effect on cardiovascular function. Clinical and epidemiological evidence suggests that the incidence of abnormal electrocardiogram (ECG) is significantly higher in Mn exposed workers than that in the control subjects. The main types of abnormal ECG include sinus tachycardia, sinus bradycardia, sinus arrhythmia, sinister megacardia, and ST-T changes. The accelerated heart-beat and shortened P-R interval appear to be more prominent in female exposed workers than in their male counterparts. Mn-exposed workers display a mean diastolic blood pressure that is significantly lower than that of the control subjects, especially in the young and female exposed workers. Animal studies indicate that Mn is capable of quickly accumulating in heart tissue, resulting in acute or subacute cardiovascular disorders, such as acute cardiodepression and hypotension. These toxic outcomes appear to be associated with Mn-induced mitochondrial damage and interaction with the calcium channel in the cardiovascular system. PMID- 16382173 TI - Relative toxicity of cardiotonic agents: some induce more cardiac and skeletal myocyte apoptosis and necrosis in vivo than others. AB - We sought to determine the relative myotoxicity of a sample of cardiotonic (catecholaminergic and PDE Inhibitory) agents currently available for clinical use. Male Wistar rats (292 +/- 24 g) were administered single subcutaneous injections of 20 mmol kg(-1) of each agent. Myocyte apoptosis (caspase-3 and annexin-V) and necrosis (anti-myosin antibody) were detected immunohistochemically on cryosections of the heart and soleus muscle. All of the cardiotonic agents except dopamine produced significant amounts of cardiomyocyte death compared with the vehicle controls, with necrosis (range 2-8%, p < 0.01) approximately one order of magnitude greater in extent than apoptosis (range 0.06 0.5%, p < 0.05). The incidence of necrosis induced by norepinephrine (8%) was approximately twice that of epinephrine and isoproterenol (4 %) and four times that of dobutamine and milrinone (2%). All agents were also toxic to the soleus muscle (range 0.1-8%), but isoproterenol (8%, p < 0.05) and epinephrine (4%, p < 0.05) were the most significant. No cell death was detected in control animals treated with only the vehicle. A majority of cardiotonic agents currently in clinical use are toxic to cardiac and skeletal myocytes. These observations suggest that judicious clinical use of such agents requires careful weighing of potential benefits against the harm via accelerated cumulative loss of myocytes. PMID- 16382174 TI - Reduction of 13-deoxydoxorubicin and daunorubicinol anthraquinones by human carbonyl reductase. AB - Carbonyl reductase (CR) catalyzes the nicotinamide adenine dinucleotide phosphate (NADPH)-dependent reduction of several carbonyls. Anthracyclines used to treat cancer are reduced by CR at the C13 carbonyl and the resulting metabolites are implicated in the cardiotoxicity associated with anthracycline therapy. CR also is believed to have a role in detoxifying quinones, raising the question whether CR catalyzes reduction of anthracycline quinones. Steady-state kinetic studies were done with several anthraquinone-containing compounds, including 13 deoxydoxorubicin and daunorubicinol, which lack the C13 carbonyl, thus unmasking the anthraquinone for study. k(cat) and k(cat)/K(m) values for 13 deoxydoxorubicin and daunorubicinol were nearly identical, indicating that that the efficiency of quinone reduction was unaffected by the differences at the C13 position. k(cat) and k(cat)/K(m) values were much smaller for the analogs than for the parent compounds, suggesting that the C13 carbonyl is preferred as a substrate over the quinone. CR also reduced structurally related quinone molecules with less favorable catalytic efficiency. Modeling studies with doxorubicin and carbonyl reductase revealed that methionine 234 sterically hinder the rings adjacent to the quinone, thus accounting for the lower catalytic efficiency. Reduction of the anthraquinones may further define the role of CR in anthracycline metabolism and may influence anthracycline cytotoxic and cardiotoxic mechanisms. PMID- 16382175 TI - Cocaine increases intracellular calcium and reactive oxygen species, depolarizes mitochondria, and activates genes associated with heart failure and remodeling. AB - To determine the cardiovascular molecular events associated with acute exposure to cocaine, the present study utilized in vivo analysis of left-ventricular heart function in adult rabbits, fluorescence confocal microscopy of fluo-2, rhod-2, (5 (and-6) carboxy 2',7' dichlorodihydrofluores-cein diacetate (carboxy-H2DCFDA), and JC-1 in H9C2 cells and gene expression microarray technology for analysis of gene activation in both rabbit ventricular tissue and H9C2 cells. In the rabbit, acute cocaine exposure (2 mg/kg) caused left-ventricular dysfunction and 0.1-10 mM cocaine increased cytosolic and mitochondrial calcium activity and mitochondrial membrane depolarization in H9C2 cells. A 3-min pretreatment of H9C2 cells by 10 microM verapamil, nifedipine, or nadolol inhibited calcium increases, but only 1 mM N-acetylcysteine (NAC) or 1 mM glutathione blocked mitochondrial membrane depolarization. Cocaine induced activation of genes in the rabbit heart and H9C2 cells including angiotensinogen, ADRB1, and c-reactive protein (CRP). In H9C2 cells, NAC pretreatment blocked cocaine-mediated increases in CRP, FAS, FAS ligand, and cytokine receptor-like factor1 (CRLF1) expression. Collectively, these data suggest that acute cocaine administration initiates cellular and genetic changes that, if chronically manifested, could cause cardiac deficits similar to those seen in heart failure and ischemia, such as ventricular dysfunction, cardiac arrhythmias, and cardiac remodeling. PMID- 16382176 TI - Role of vagal activity on bradicardic and hypotensive effects of caffeic acid phenethyl ester (CAPE). AB - Caffeic acid phenethyl ester (CAPE) is a phenolic active component of propolis of honeybee hives and reduces heart rate and blood pressure in rats. The objective of this study was to investigate the role of vagal activity and atropine blockage on the bradycardic and hypotensive effects of CAPE in rats. The rats were divided into five groups (n = 8). Saline and vehicle (10% ethanol) of CAPE were given to the first and second groups, respectively. Group 3 was treated with 5 mg/kg CAPE. Group 4 bivagotomized and treated with 5 mg/kg CAPE. Group 5 treated with atropine (5 microg/microL/min) continuously and treated with CAPE. The electrophysiological monitoring was done for each experiment under urethane anesthetize. As a result, CAPE caused intense and transient bradycardia and hypotension. Vagotomy completely abolished bradycardia occurred via CAPE injection; however atropine attenuated bradycardic effects of CAPE. On the other hand, hypotensive effect of CAPE was affected from neither bilateral vagotomy nor atropine treatment. It was thought that CAPE may exert its effects on heart rate via a central parasympathetic control mechanism, but not on central parasympathetic blood pressure control system. PMID- 16382178 TI - A pull-down assay for 5' AMP-activated protein kinase activity using the GST fused protein. AB - An assay using a specific peptide (SAMS peptide) as a substrate is widely used for determination of AMP-activated protein kinases (AMPK) activity. However, it is not an efficient assay for crude AMPK preparations. In this study, we modified the assay by using the SAMS peptide fused to glutathione-S-transferase (GST-SAMS) instead of the SAMS peptide on its own. Radioactivity incorporated into GST-SAMS can be recovered easily by precipitation with glutathione-agarose. The kinetic parameters of partially purified AMPK for the GST-SAMS were as follows. The Vmax was 0.26 +/- 0.012 nmol/min/mg of total proteins and Km for GST-SAMS was 110 +/- 12 microM. The parameters for ATP were 0.40 +/- 0.016 nmol/min/mg of total proteins (Vmax) and 202 +/- 21 microM (Km). The activity of AMPK in this system was stimulated about threefold by the AMPK activators, AMP or 5-amino-4 imidazolecarboxamide ribotide (ZMP), and inhibited by the AMPK inhibitors, adenine 9-beta-D-arabinofuranoside (ara-A) and iodotubercidin. These values correlate well with those for the SAMS peptide reported previously. Thus, we successfully established a convenient and rapid method to measure AMPK applicable, even for crude enzyme preparations. PMID- 16382177 TI - Bacterial and plant enterotoxin B subunit-autoantigen fusion proteins suppress diabetes insulitis. AB - Several bacterial and plant enterotoxin B subunit-islet autoantigen fusion proteins were compared for their ability to serve as islet autoantigen carriers and adjuvants for reduction of pancreatic islet inflammation associated with type 1 diabetes. The cholera toxin B subunit (CTB), the heat-labile toxin B subunit from enterotoxigenic Escherichia coli (LTB), the Shigella toxin B subunit (STB), and the plant toxin ricin B subunit (RTB) were genetically linked to the islet autoantigens proinsulin (INS) and glutamic acid decarboxylase (GAD). The adjuvant autoantigen gene fusions were transferred to a bacterial expression vector and the corresponding fusion proteins synthesized in E. coli. The purified adjuvant autoantigen proteins were fed to 5-wk-old nonobese diabetic (NOD) mice once a week for 4 wk. Histological examination of pancreatic islets isolated from inoculated mice showed significant levels of insulitis reduction in comparison with uninoculated mice. The ratio of serum anti-INS and anti-GAD IgG2c to IgG1 antibody isotype titers increased in all ligand-autoantigen inoculated animal groups, suggesting an increase in effector Th2 lymphocytes in B subunit-mediated insulitis suppression. The results of these experiments indicate that bacterial and plant enterotoxin B subunit ligand-autoantigens enhance insulitis reduction in NOD mice. This research prompts further exploration of a multiadjuvant/autoantigen co-delivery strategy that may facilitate type 1 diabetes prevention and suppression in animals and humans. PMID- 16382179 TI - Quantitative competitive-PCR assay to measure human parvovirus B19-DNA load in serum samples. AB - The B19 virus can persist in immunocompromised patients for several months and sometimes even years because of impaired immune response. Viremia in persistent and recurrent infection may range from very low to high titers and may be associated with chronic clinical manifestations, such as chronic anemia. Several recently developed techniques that quantify B19-DNA have improved laboratory diagnosis of the infection and can help guide the choice of treatment in persistent infections (i.e., intravenous immunoglobulin (IVIG) treatment vs immunosuppression reduction). Here we describe the development of a reliable internally controlled quantitative competitive (QC)- polymerase chain reaction (PCR) assay that measures B19-DNA load in serum samples by densitometric analysis of the amplification products for monitoring B19 infection in high-risk patients. A retrospective quantification of B19-DNA in the serum samples from 48 anemic transplanted patients by the QC-PCR assay we developed in our laboratory confirmed the presence of B19-DNA in 11 of 48 samples and showed a viral DNA load between 103 and 108 B19-DNA copies/mL depending on the patients' serostatus (the highest viral load was found in IgM-positive/IgG-negative patients, that is, in patients with active B19 infection at onset). The assay also confirmed B19-DNA negative patients. Our QC-PCR assay may be easily used in monitoring B19 prototype DNA load to follow persistent infections and to better understand the relation between active B19 infection and occurrence of anemia and to assess the efficacy of IVIG therapy or immunosuppression reduction in clearing the virus in high-risk patients. PMID- 16382180 TI - Analysis of immune response in young and aged mice vaccinated with corn-derived antigen against Escherichia coli heat-labile enterotoxin. AB - Enterotoxigenic strains of Escherichia coli produce a heat-labile holotoxin (LT), which causes diarrhea. We engineered corn seeds to produce LT-B, the nontoxic subunit of LT, to serve as a plant-derived vaccine to traveler's diarrhea and as an adjuvant for co-administered proteins. We previously demonstrated that a strong mucosal and systemic antibody response is elicited in young mice with oral administration of corn-derived LT-B. The present study examined systemic and mucosal antibody responses to LT-B in young and aged mice, and recall responses to oral administration and injection of LT-B in aged mice. Specific IgA and IgG antibodies were detectable during an 11-mo period, although the concentration of antigen-specific antibodies declined gradually. Booster by feeding or injection dramatically increased the concentration of specific IgA from that seen in young mice. Specific IgG levels were boosted to concentrations similar to those in young mice. This effect may be age-dependent and related to prior immunization exposure. Analysis of the antibody response of naive aged mice against corn derived LT-B demonstrated an age-related suppression in specific IgG production, but not specific IgA. These results may provide important information for edible vaccine strategies for young and aged individuals. PMID- 16382181 TI - An improved recombineering approach by adding RecA to lambda Red recombination. AB - Recombineering is the use of homologous recombination in Escherichia coli for DNA engineering. Of several approaches, use of the lambda phage Red operon is emerging as the most reliable and flexible. The Red operon includes three components: Redalpha, a 5' to 3' exonuclease, Redbeta, an annealing protein, and Redgamma, an inhibitor of the major E. coli exonuclease and recombination complex, RecBCD. Most E. coli cloning hosts are recA deficient to eliminate recombination and therefore enhance the stability of cloned DNAs. However, loss of RecA also impairs general cellular integrity. Here we report that transient RecA co-expression enhances the total number of successful recombinations in bacterial artificial chromosomes (BACs), mostly because the E. coli host is more able to survive the stresses of DNA transformation procedures. We combined this practical improvement with the advantages of a temperature-sensitive version of the low copy pSC101 plasmid to develop a protocol that is convenient and more efficient than any recombineering procedure, for use of either double- or single stranded DNA, published to date. PMID- 16382182 TI - Site-specific DNA excision in transgenic rice with a cell-permeable cre recombinase. AB - The removal of selected marker genes from transgenic plants is necessary to address biosafety concerns and to carry out further experiments with transgenic organisms. In the present study, the 12-amino-acid membrane translocation sequence (MTS) from the Kaposi fibroblast growth factor (FGF)-4 was used as a carrier to deliver enzymatically active Cre proteins into living plant cells, and to produce a site-specific DNA excision in transgenic rice plants. The process, which made cells permeable to Cre recombinase-mediated DNA recombination, circumvented the need to express Cre under spatiotemporal control and was proved to be a simple and efficient system to achieve marker-free transgenic plants. The ultimate aim of the present study is to develop commercial rice cultivars free from selected marker genes to hasten public acceptance of transgenic crops. PMID- 16382183 TI - Megaprimer-based methodology for deletion of a large fragment within a repetitive polypyrimidine-rich DNA. AB - Site-directed mutagenesis is often a prerequisite for elucidation of the functional significance of cis- and trans-factors involved in gene regulation. The aim of this study was to delete the primary binding site for heterogeneous nuclear ribonucleoprotein I (hnRNPI) within the inducible nitric oxide synthase (iNOS) 3' untranslated region mRNA. The binding site consists of a 53-nucleotide CU-rich region within a long stretch of polypyrimidines. As a result of primer pair annealing, the repetitive sequence limited the use of several deletion methods based on polymerase chain reaction. Therefore, a megaprimer approach was chosen. The megaprimer was produced by a forward primer outside the polypyrimidine-rich region, and a mutagenic reverse primer annealing to flanking regions of the desired deletion, thereby looping out the target sequence. Subsequently, this megaprimer was used to create the final deletion recombinant. The deletion was verified by sequencing and by ultraviolet cross-linking mouse liver protein extracts with radiolabeled mutant and wild-type RNAs. In conclusion, the megaprimer method offers a solution for generating large internal deletions in repetitive sequences, which facilitates investigations on large repetitive DNA or RNA regions interacting with trans-factors. PMID- 16382184 TI - Secretion of pro- and mature Rhizopus arrhizus lipases by Pichia pastoris and properties of the proteins. AB - The lipases of Rhizopus spp. share a high 1,3-regiospecificity toward triacylglycerols, which makes them important enzymes in lipid modification. In the present study, the extracellularly active production of recombinant Rhizopus arrhizuslipase was carried out with genes encoding the mature region (mRAL) and the mRAL having the prosequence (ProRAL) in Pichia pastoris. Two transformed P. pastoris clones containing the multicopy of mRAL and ProRAL genes were separately selected for the production of recombinant enzymes. In a fed-batch cultivation, where methanol feeding was controlled by an on-line methanol analyzer, the supernatant contained 91 mg/L recombinant pro-form lipase (rProRAL) and 80 mg/L recombinant mature lipase (rRAL) after 92 h of cultivation. rProRAL and rRAL were purified by ultrafiltration, SP-Sepharose Fast Flow chromatography, and Butyl Sepharose Fast Flow chromatography. Molecular weights of rProRAL and rRAL are 32 kDa and 29 kDa, respectively. The amino-terminal analysis showed that the 32-kDa protein was mRAL attached with 28 amino acids of the carboxy-terminal part of the prosequence (rPro28RAL). The specific lipase activities of mRAL attached with 28 amino acids of the carboxy-terminal part of the prosequence (rPro28RAL) and rRAL were 1543 U/mg and 2437 U/mg. The rPro28RAL was more stable than rRAL at pH 4.0 7.0, whereas rRAL was more stable at pH 7.0-10.0. The rPro28RAL had the highest lipase activity toward tributyrin (C4), whereas rRAL had the highest lipase activity toward tricaprylin (C8). PMID- 16382185 TI - Isolation of C. elegans deletion mutants following ENU mutagenesis and thermostable restriction enzyme PCR screening. AB - The ability to generate null mutants is essential for studying gene function. Gene knockouts in Caenorhabditis elegans can be generated in a high throughput manner using chemical mutagenesis followed by polymerase chain reaction (PCR) assays to detect deletions in a gene of interest. However, current methods for identifying deletions are time and labor intensive and are unable to efficiently detect small deletions. In this study, we expanded the method pioneered by Wei et al., which used the thermostable restriction enzyme PspGI and tested the usefulness of other thermostable restriction enzymes including BstUI, Tsp45I, ApeKI, and TfiI. We designed primers to flank one or multiple thermostable restriction enzymes sites in the genes of interest. The use of multiple enzymes and the optimization of PCR primer design enabled us to isolate deletion in 66.7% of the genes screened. The size of the deletions varied from 330 bp to 1 kb. This method should make it possible for small academic laboratories to rapidly isolate deletions in their genes of interest. PMID- 16382187 TI - The serotonergic system and its role in cocaine addiction. AB - Cocaine is an alkaloid with psychostimulant action and high addictive potential. It possesses high affinity for the transporters of dopamine, serotonin (5-HT) and noradrenaline, and blocks reuptake of the above-mentioned monoamines. The present review summarized the contribution of 5-HT neurotransmitter system to rewarding and aversive properties of cocaine, to cocaine withdrawal and its long-term abuse. The present state of knowledge of 5-HT neurotransmission justifies the opinion that pharmacological manipulation in the 5-HT system may efficiently counteract the effects of cocaine withdrawal and prevent reinstatement of its abuse. PMID- 16382188 TI - Short review on dopamine agonists: insight into clinical and research studies relevant to Parkinson's disease. AB - Parkinson's disease (PD) is a chronic and progressive neurological disorder characterized by selective degeneration of dopaminergic neurons (DAergic) in the substantia nigra pars compacta (SNpc) and subsequent decrease in dopamine (DA) levels in the striatum. Although levodopa replacement therapy is initially effective in symptomatic treatment of parkinsonian patients, its effectiveness often declines and various levodopa-related side effects appear after long-term treatment. The disabling side effects of levodopa therapy include motor fluctuations such as the wearing-off or on-off phenomena, dyskinesias and psychiatric symptoms. Nowadays, DA receptor agonists are often regarded as first choice in de novo and young parkinsonian patients to delay the onset of levodopa therapy. In advanced stages of the disease, they are also used as adjunct therapy together with levodopa to retard the development of motor complications. DA receptor agonists mimic the endogenous neurotransmitter, dopamine, and act by direct stimulation of presynaptic (autoreceptors) and postsynaptic DA receptors. Next to their clinical role in treating parkinsonian patients, laboratory studies reported antioxidative and neuron-rescuing effects of DA receptor agonists either in vivo or in vitro. This may involve reduced DA turnover following autoreceptor stimulation and direct free radical scavenging activity. In this review, we focus on and summarize the recently reported effects of the most commonly used DA agonists either in clinical or in research studies relevant to PD treatment. PMID- 16382189 TI - Zinc and depression. An update. AB - Unsatisfactory clinical efficacy and a variety of adverse effects of current antidepressant drugs have incited search for better therapy. Zinc, an antagonist of the glutamate/N-methyl-D-aspartate (NMDA) receptor, exhibits antidepressant like activity in rodent tests/models of depression. Similarly to antidepressants, zinc induces brain derived neurotrophic factor (BDNF) gene expression and increases level of synaptic pool of zinc in the hippocampus. Clinical observations demonstrated serum hypozincemia in depression, which was normalized by effective antidepressant treatment. Moreover, our preliminary clinical study demonstrated the benefit of zinc supplementation in antidepressant therapy. All the data indicate the important role of zinc homeostasis in psychopathology and therapy of depression and potential clinical antidepressant activity of this ion. PMID- 16382190 TI - Profile of anticonvulsant activity and neuroprotective effects of novel and potential antiepileptic drugs--an update. AB - Although neuroprotection is effective only against certain aspects of a complex cascade of pathological events during the development and course of epilepsy, it might be a promising option in the treatment of this disease. Some new data on the pathophysiology of epilepsy raised some hopes that the epileptogenesis process can be prevented. A question arises whether it is possible to make the epilepsy develop in a milder, easier to treat and non-progressive way without cognitive decline and drug-resistance. Moreover, once the epilepsy has already been triggered, there is as yet no conclusive evidence that the harmful effects of seizures on the brain can be reduced. So a great deal of further evaluation of antiepileptic drugs (AEDs) is required. Many similarities exist between cerebral ischemia and epilepsy regarding brain-damaging and autoprotective mechanisms that are activated following the injurious insult. Therefore, drugs that are effective in minimizing seizure-induced brain damage may also be useful in minimizing ischemic injury. Most AEDs have been tested in animal models of focal or global ischemia and some were already tested in humans for a possible neuroprotective effect. The existing data are rather scanty and insufficient but it appears that only drugs that have multiple mechanisms of action have some potential in conferring a degree of neuroprotection that could be clinically applicable to stroke patients. In this review, we focus on evidence of neuroprotective properties of novel and potential AEDs, based on animal experimental models of neurodegeneration. In conclusion, some of the newer AEDs show promise as possible neuroprotectants in epilepsy and acute ischemia but more studies are needed before clinical trials in humans could be undertaken. PMID- 16382191 TI - Anxiolytic action of group II and III metabotropic glutamate receptors agonists involves neuropeptide Y in the amygdala. AB - Several lines of evidence indicate that activation of group II and III metabotropic glutamate (mGlu) receptors produces anxiolytic-like effects in rodents. On the other hand neuropeptide Y (NPY) induces an anxiolytic effect in rats after intraventricular or intraamygdalar administration. Therefore, in the present study we investigated whether the anxiolytic action of (2S,3S,4S) (carboxycyclopropyl)glycine (L-CCG-I), an mGlu2/3 receptor agonist, and (1S,3R,4S)-1-aminocyclopentane-1,3,4-tricarboxylic acid (ACPT-1), an mGluR4/6/7/8 receptor agonist, was mediated by a mechanism involving NPY receptor. In behavioral studies, the anxiolytic activity of L-CCG-I (10 microg/0.5 microl/site) and ACPT-1 (1.5 microg/0.5 microl/site) was examined using plus-maze tests. The Y1 receptor antagonist BIBO 3304 was given at a dose of 128 ng/0.5 microl/site. All the compounds tested were injected bilaterally into the amygdala, BIBO 40 min and mGluR agonists 30 min before the test. It was found that the anxiolytic effects of mGluR agonists were abolished by BIBO 3304 {((R)-N [[4-(aminocarbonylaminomethyl) phenyl] methyl]-N2-(diphenylacetyl)-argininamide trifluoroacetate)3304} administration. Immunohistochemical studies showed a moderate density of mGlu2/3 receptor immunoreactivity (IR) in the amygdala. The effect of L-CCG-I and ACPT-1 on NPY expression in the amygdala was studied using immunohistochemistry (IH), while NPYmRNA expression was studied using in situ hybrydization. We showed a diminution in NPY-IR after L-CCG-I administration and decrease in NPYmRNA expression after both L-CCG-I and ACPT-1 treatment, to about 77% (IH) or 32-41% (mRNA) of the control level 18 h after injection of these mGluR agonists. Our results indicate that the anxiolytic action of both compounds is conveyed by NPY neurons with the involvement of Y1 receptors in the amygdala, and that NPY neurons seem to be regulated by the glutamatergic system. PMID- 16382192 TI - Role of opioidergic mechanisms and GABA uptake inhibition in the heroin-induced discriminative stimulus effects in rats. AB - The present study was designed to investigate the involvement of opioidergic component as well as to study GABAergic mechanisms in the expression of heroin discrimination. Male Wistar rats were trained to discriminate heroin (0.5 mg/kg, i.p.) from saline (i.p.) in a two-choice water reinforced fixed ratio (FR) 20 drug discrimination paradigm. In substitution tests, heroin (0.0625-0.5 mg/kg) and morphine (0.5-2 mg/kg, i.p.) evoked a dose-dependent generalization for the drug lever-responding, while muscimol (1 mg/kg, i.p., a GABA(A) receptor agonist) produced a weak partial substitution (ca. 48% heroin-lever responding). Neither tiagabine (2.5 mg/kg, i.p.; a GABA reuptake inhibitor), vigabatrin (75-150 mg/kg, i.p.; an irreversible inhibitor of GABA transaminase), nor baclofen (0.5 mg/kg, i.p.; a GABA(B) receptor agonist) substituted for heroin. In combination studies, the stimulus effects produced by heroin (0.5 mg/kg) or morphine (2 mg/kg) were dose-dependently blocked by opioid receptor antagonists naltrexone (0.1-1 mg/kg, i.p.), and naloxone (0.5-1 mg/kg, i.p.). The peripherally-acting naloxone methiodide at a dose of 1 mg/kg, i.p. did not alter, while at a dose of 10 mg/kg that penetrates across the blood-brain barrier, it reduced the stimulus effects of heroin or morphine. Pretreatment with tiagabine (2.5-5 mg/kg) produced a rightward shift of a heroin dose-response curve, while vigabatrin (75-300 mg/kg), baclofen (0.5-2.5 mg/kg) or muscimol (0.5-2 mg/kg) given prior to heroin (0.0625 0.5 mg/kg) failed to alter heroin discrimination. Our findings confirm previous studies on the significance of mu-opioidergic mechanisms in the expression of heroin discrimination and add the observation that selective inhibition of GABA reuptake, but not inhibition of GABA transaminase or direct stimulation of GABA(A) and GABA(B) receptors, can decrease the overall effects of heroin. PMID- 16382193 TI - Naloxone precipitates nicotine abstinence syndrome and attenuates nicotine induced antinociception in mice. AB - The present study focused on the evaluation of a role of opioid system in nicotine-induced antinociception and physical dependence in mice. The results indicate that nicotine (3 mg/kg) produced a significant antinociception in the hot plate test. Additionally, the opioid receptor antagonist naloxone (0.5 and 1 mg/kg), dose-dependently attenuated this effect. Our second experimental protocol consisted in intermittent administration of nicotine (2.5 mg/kg, s.c.) four times daily for 7 days. In order to precipitate nicotine abstinence, mice were given one injection of mecamylamine (3 mg/kg) or naloxone (1 mg/kg) one hour after the last nicotine injection on the test day (day 8) in the morning. Interestingly, our findings revealed that both drugs precipitated somatic withdrawal signs in mice, with a slight difference in their influences on the intensity of several signs. These data support the hypothesis that similar opioid-cholinergic interactions are involved in nicotine-induced antinociception and nicotine withdrawal syndrome. PMID- 16382194 TI - Association studies of 5-HT2A and 5-HT2C serotonin receptor gene polymorphisms with prophylactic lithium response in bipolar patients. AB - Lithium is one of the most commonly used drugs in the prophylaxis and treatment of bipolar disorder. The mechanisms of mood stabilization by lithium incorporate its effect on serotonergic neurotransmission. This paper investigates a relationship between response to lithium prophylaxis and polymorphisms in two genes: T102C of 5-HT2A receptor and G68C (Cys23Ser) of 5-HT2C serotonin receptor gene. Genotypes were estimated in 92 bipolar patients (39 males and 53 females) who have been taking lithium for at least 5 years. The patients were classified as excellent responders, partial responders and non-responders to lithium. The obtained results suggest that these polymorphisms may not be related to the degree of prophylactic lithium response. PMID- 16382195 TI - Effects of some new antidepressant drugs on the glucocorticoid receptor-mediated gene transcription in fibroblast cells. AB - Antidepressant drugs are thought to counteract effects of hypercortisolemia, frequently associated with depression, by lowering cortisol level and by modifying the function of glucocorticoid receptors (GR). Indeed, classical antidepressants inhibit corticosteroid-induced gene transcription in cell cultures. The aim of the present study was to investigate effects of new generation antidepressant drugs on GR function in mouse fibroblast cells (L929), stably transfected with mouse mammary tumor virus-chloramphenicol acetyltransferase (MMTV-CAT) plasmid (LMCAT cells). It has been found that reboxetine (at 10 and 30 microM), venlafaxine, citalopram and mirtazapine (at 30 microM), but not milnacipran, in statistically significant manner inhibited corticosterone-induced gene transcription. However, the effects of new generation antidepressant drugs were weaker than those evoked by imipramine, which was active already at 3 microM concentration. Further studies on the mechanism of antidepressant action on GR function revealed that protein kinase C, but not mitogen-activated protein kinases (MAPK), glycogen synthase kinase (GSK-3) and protein kinase B (PKB, Akt) play a role in this phenomenon. PMID- 16382197 TI - How significant is the difference between drug doses influencing the threshold for electroconvulsions? AB - In this study, three computational methods (log-probit, one-way analysis of variance and linear regression analysis) were applied to statistically determine and compare a minimal dose of the antiepileptic drug--levetiracetam (LEV), which significantly increased the threshold for maximal electroconvulsions in mice. Results analyzed with log-probit method revealed that such a dose for LEV was 50 mg/kg. In contrast, one-way ANOVA indicated that the minimal dose of LEV, fulfilling the criterion of the significant increment in the threshold for electroconvulsions, was 100 mg/kg. Moreover, linear regression analysis allowed for the determination of threshold increasing doses by 20% (TID20) and 50% (TID50) for LEV, which amounted to 44 mg/kg and 150 mg/kg. This comparative study demonstrated distinct differences between the electroconvulsive threshold influencing doses of LEV calculated with these three methods, and only statistical evaluation of data using one-way ANOVA provided the most adequate determination of statistical significance. The application of log-probit method and linear regression analysis in the search for significant drug-dose effects should be substantially restricted to specific experimental conditions. PMID- 16382196 TI - Effect of short- and long-term treatment with antidepressant drugs on the activity of rat CYP2A in the liver. AB - The aim of the present study was to investigate the influence of tricyclic antidepressants (TADs: imipramine, amitriptyline, clomipramine, desipramine), selective serotonin reuptake inhibitors (SSRIs: fluoxetine, sertraline) and novel antidepressant drugs (mirtazapine, nefazodone) on the activity of CYP2A measured as a rate of testosterone 7alpha-hydroxylation. The reaction was studied in control liver microsomes in the presence of the antidepressants, as well as in microsomes of rats treated intraperitoneally (i.p.) for one day or two weeks with pharmacological doses of the drugs (imipramine, amitriptyline, clomipramine, nefazodone 10 mg/kg i.p.; desipramine, fluoxetine, sertraline 5 mg/kg i.p.; mirtazapine 3 mg/kg i.p.), in the absence of the antidepressants in vitro. Most of the investigated drugs directly inhibited the CYP2A activity when added in vitro to control liver microsomes. Their inhibitory effects were strong (clomipramine, fluoxetine and desipramine: Ki = 15, 20 and 25 microM, respectively), moderate (sertraline and imipramine: Ki = 50 and 75 microM, respectively) or weak (amitriptyline, nefazodone and mirtazapine: Ki = 107, 127 and 250 microM, respectively). A one-day (i.e. 24-h) exposure to the investigated antidepressant drugs did not produce any significant changes in the rate of 7alpha-hydroxylation of testosterone in the rat liver microsomes, while chronic treatment with clomipramine or sertraline significantly increased the activity of CYP2A, which suggests enzyme induction. In summary, two different mechanisms of the antidepressant-CYP2A interaction have been found in rat liver: 1) the direct inhibition of CYP2A by most of the investigated TADs and SSRIs; 2) the in vivo weak induction of CYP2A by clomipramine and sertraline. This observation may be important to the interpretation of the results of pharmacological tests carried out on rats. It seems of primary importance to determine whether the influence of antidepressants on CYP2A6 in humans is analogous as on CYP2A1/2 in rats. PMID- 16382198 TI - 2-Chloro-N6-cyclopentyladenosine enhances the anticonvulsant action of carbamazepine in the mouse maximal electroshock-induced seizure model. AB - This study examines the anticonvulsant profile of interactions between 2-chloro N6-cyclopentyladenosine (CCPA, a selective adenosine A1 receptor agonist) and four conventional antiepileptic drugs (AEDs: carbamazepine--CBZ, phenobarbital, phenytoin and valproate) in the mouse maximal electroshock seizure (MES) model. Acute adverse effects produced by AEDs in combination with CCPA were determined in the chimney test (motor performance) and passive avoidance task (long-term memory). Results indicate that CCPA administered alone at 0.25 and 0.5 mg/kg significantly elevated the electroconvulsive threshold in mice. Additionally, the agent at a sub-threshold dose of 0.125 mg/kg potentiated the anticonvulsant activity of CBZ by reducing its ED50 in the MES test from 11.2 to 7.7 mg/kg (p < 0.01). In contrast, 8-cyclopentyl-1,3-dimethylxanthine (DPCPX, a selective adenosine A1 receptor antagonist at 5 mg/kg) abolished the enhanced anticonvulsant effects offered by the combination of CBZ with CCPA (0.125 mg/kg). Moreover, CCPA (0.125 mg/kg) co-administered with other tested AEDs had no significant impact on their antiseizure properties in the MES test in mice. Neither CCPA (0.125 mg/kg) administered singly, nor in combinations with conventional AEDs (at their ED50s) affected motor performance in the chimney test and long-term memory in the passive avoidance task. No pharmacokinetic alterations in brain CBZ concentrations were observed after administration of CCPA at 0.125 mg/kg. It may be concluded that CCPA, acting selectively on adenosine A1 receptors, enhances pharmacodynamically the antiseizure effect of CBZ in the MES test. PMID- 16382199 TI - Non-competitive metabotropic glutamate subtype 5 receptor antagonist (SIB-1893) decreases body temperature in rats. AB - This study examined the effect of (E)-2-methyl-6-(2-phenylethynyl)-pyridine (SIB 1893), a selective non-competitive metabotropic glutamate subtype 5 receptor (mGluR5) antagonist, on body temperature in freely moving Wistar rats. Temperature was monitored using programmed microchips, implanted subcutaneously in rats, at several times: 0, 5, 10, 20, 30, 45, 60, 90, 120 and 180 min after intraperitoneal administration of SIB-1893 at increasing doses of 10, 20 and 30 mg/kg. The results analyzed with two-way ANOVA with repeated measures on time revealed that SIB-1893 at 30 mg/kg considerably lowered the body temperature in animals at 90, 120 and 180 min after its systemic injection. In contrast, the drug at 10 and 20 mg/kg remained without effect on the body temperature in rats. Based on our preclinical study, one can conclude that SIB-1893 produces hypothermia in freely moving rats in a dose-dependent manner. PMID- 16382200 TI - Analgesic and anti-inflammatory activity of stereoisomers of carane derivatives in rodent test. AB - Our previously conducted pharmacological investigations led us to discovery of the strong local anesthetic activity of the compound KP-23RS. The following studies revealed that its R- and S-diasteroisomers had different activity in the local anesthetic, anti-aggregating, anti-arrhythmic and spasmolytic tests. Also the influence of KP-23RS and its diastereoisomers on the cyclic adenosine monophosphate (AMP) generating system was described. In the present study, anti inflammatory and analgesic effects of these compounds were investigated in hind paw edema test, Randall's analgesia test and hot-plate test. Also the spasmolytic activity and the influence on the stomach mucous membrane were examined. All of these compounds had an anti-inflammatory and analgesic activity in hot-plate test and in Randall's test. Moreover, compound KP-23R showed spasmolytic activity. None of the investigated compounds induced damage of the mucous membrane of the rat stomach. PMID- 16382201 TI - Effects of lactoferrin on the immune response modified by the immobilization stress. AB - Effects of orally administered lactoferrin (LF) on the cellular and humoral immune responses in mice subjected to immobilization stress (IS) were investigated. Here, we demonstrate that long-term IS induced significant suppression of cellular and humoral immune responses in CBAmice. The suppression was attenuated by LF given to mice in drinking water as determined by the number of antibody-forming cells (AFC) in the spleen and the magnitude of delayed type of hypersensitivity (DTH). On the other hand, LF lowered the elevated DTH response in mice exposed to short-term IS (5 h only) on the day of elicitation of the DTH reaction. We also showed that LF up-regulated spontaneous transforming growth factor beta (TGF-beta) production in the cultures of mesenteric lymph node cells derived from short-term stressed mice. This is the first report on the regulatory effect of LF on the immune response modified by the psychic stress and is consistent with other reports on antinociceptive and analgesic actions of LF in experimental animals. PMID- 16382202 TI - Microarray analysis of altered gene expression in diallyl trisulfide-treated HepG2 cells. AB - Diallyl trisulfide (DT) is a natural compound derived from garlic. Despite its reported lipid-lowering effects, the mechanisms of its actions are not clear yet. To further understand the molecular mechanisms of actions of this compound, microarray technology was used in the present study to investigate the lipid lowering effects of DT on HepG2 cells. To optimize the concentration of DT treatment on HepG2 cells, a series of concentration of DT were incubated with HepG2 cells for 24 h. The data indicted that the concentrations of DT in the range from 20 to 50 microM were effective in lowering cellular total triglyceride and cholesterol, with no significant cytotoxicity. Using oligonucleotide microarrays and RT-PCR technology, the genes that were differentially regulated by DT were identified. The results showed that peroxisome proliferator activated receptor alpha (PPAR-alpha) and hepatocyte nuclear factor 4alpha (HNF-4alpha) mRNA were up-regulated, and CYP7A1 mRNA was down-regulated following DT treatment, suggesting that the lipid-lowering effects of DT may be at least in part mediated through the regulation of PPAR-alpha dependent pathways. PMID- 16382203 TI - Isolated rat inferior mesenteric artery response to adenosine: possible participation of Na+/K+-ATPase and potassium channels. AB - Adenosine (10(-7)-3 x 10(-4) M) produced concentration-dependent and endothelium independent relaxation of isolated rat inferior mesenteric artery. Application of indomethacin (10(-5) M) or N(G)-nitro-L-arginine (10(-5) M) did did not alter adenosine-elicited relaxation. Conversely, in the presence of high concentration of K+ (100 mM), ouabain (10(-4)) or combination of tetraethylammonium (5 x 10(-4) M) and glibenclamide (10(-6) M), adenosine-evoked relaxant effect was significantly reduced. In K+-free solution, 1-3 mM potassium induced relaxation, which was partially reversed by ouabain (10(-4) M). 1,3-Dipropyl-8 cyclopentylxanthine (10(-9) M), an A1-receptor antagonist, did not affect adenosine-evoked relaxation. Oppositely, 8-(3-chlorostyryl)-caffeine (3 x 10(-7) 10(-6) M), a selective A2A receptor antagonist, significantly inhibited adenosine induced relaxation in a concentration-dependent manner (pA2 = 6.74). These results indicate that in the isolated rat inferior mesenteric artery, adenosine produces endothelium-independent relaxation, which is partly induced by activation of smooth muscle adenosine A2A receptors, and further mediated by the activation of smooth muscle Na+/K+-ATPase and opening of mixed population of K+ channels. PMID- 16382204 TI - Effect of two non-steroidal anti-inflammatory drugs, aspirin and nimesulide on the D-glucose transport and disaccharide hydrolases in the intestinal brush border membrane. AB - The present study was designed to evaluate the effect of two commonly prescribed non-steroidal anti-inflammatory drugs (NSAIDs) with varying cyclooxygenase-2 (Cox 2) selectivities on enzyme activities and transport properties of rat intestinal brush border membrane (BBM). Female Wistar rats were divided into three different groups, viz: Group I (Control), Group II (aspirin-treated, 50 mg/kg) and Group III (nimesulide-treated, 10 mg/kg). At the end of 28 days of treatment, membranes were isolated from different intestinal segments of all the groups and changes in BBM-associated enzymes such as sucrase, lactase, maltase, alkaline phosphatase and the transport properties of D-glucose and its kinetics were studied. The results indicated a significant decrease in the activities of enzymes and transport of glucose in both the treatment groups as compared to the controls. Changes in Michaelis-Menten parameters, viz:--Km and Vmax and the thermodynamic parameters T(c) and E(a) were also seen which are indicative of adverse effects of these two NSAIDs in the intestinal membrane such as the membrane integrity. PMID- 16382205 TI - Combined treatment with imipramine and metyrapone induces hippocampal and cortical brain-derived neurotrophic factor gene expression in rats. AB - The problem of drug-resistant depression indicates a strong need for alternative antidepressant therapies. Recently it was shown that joint administration of imipramine (IMI) and metyrapone (MET), an inhibitor of glucocorticoid synthesis, produced a more potent "antidepressant" effect in the forced swimming test than did treatment with either drug alone. Our studies also showed that co administration of IMI and MET to drug-resistant, unipolar depressed patients effected a clinical improvement. In addition, recent studies indicated a role of the brain-derived neurotrophic factor (BDNF) in the mechanism of action of antidepressant drugs (ADs). Since the most potent effect of ADs on BDNF gene expression was found after prolonged treatment, in the present research we investigated the influence of repeated treatment with IMI (5 or 10 mg/kg) and MET (50 mg/kg), given separately or jointly (twice daily for 14 day), on the BDNF mRNA level (the Northern blot) in the hippocampus and cerebral cortex. The experiment was carried out on male Wistar rats. The tissue for biochemical assays was collected 24 h after the last dose of IMI and MET. The obtained results showed that in the hippocampus IMI (10 mg/kg) and cerebral cortex IMI (5 mg/kg) and MET (50 mg/kg) significantly elevated the BDNF mRNA level. Joint administration of IMI (10 mg/kg) and MET (50 mg/kg) induced a more potent increase BDNF gene expression in both the examined brain regions (compared to the treatment with either drug alone). Moreover, the obtained results suggested that BDNF may be involved in the mechanism of the synergistic antidepressant effect of IMI and MET in drug-resistant depressed patients. PMID- 16382206 TI - Purvalanol A, inhibitor of cyclin-dependent kinases attenuates proliferation of cells in the dentate gyrus of the adult rat hippocampus. AB - Several recent findings indicate that intensity of neurogenesis in adult brain might be regulated by enzymes involved in the cell cycle. Therefore, in the present study, we investigated whether the inhibition of cyclin-dependent kinases (CDKs) evoked by intraventricular administration of purvalanol A may influence the proliferation of cells in the dentate gyrus (DG) of the rat hippocampus. Purvalanol A, a selective inhibitor of CDKs, was injected into the brain lateral ventricle at concentrations of 4 nmol/3 microl or 40 nmol/3 microl. The number of proliferating cells was determined by analysis of nuclear incorporation of BrdU (100 mg/kg, i.p.). BrdU was given at two times points (0.5 h and 2.5 h) after vehicle or purvalanol A injection. It was found that purvalanol A given 0.5 h before BrdU injection did not cause any significant changes in the number of BrdU positive nuclei in the DG. However, a higher dose of purvalanol A i.e. 40 nmol/3 microl given 2.5 h before BrdU administration significantly decreased the number of BrdU-positive nuclei in the DG of the hippocampus (by approximately 35%). In contrast, a lower dose of the CDKs inhibitor (4 nmol/3 microl) given at the same time point did not affect the number of BrdU-positive cells in the DG. It appears that purvalanol A inhibits the hippocampal proliferation in concentration- and time-dependent manner. Moreover, the present data indicate that CDKs are involved in generation of new cells in the adult rat hippocampus. PMID- 16382208 TI - Lack of the antianxiety-like effect of (S)-3,4-DCPG, an mGlu8 receptor agonist, after central administration in rats. AB - Substances acting as agonists of group III mGlu receptors were shown to induce an antianxiety-like effect after intrahippocampal administration to rats. The purpose of the present study was to establish whether the selective mGlu8 receptor agonist (S)-3,4-dicarboxyphenylglycine ((S)-3,4-DCPG) induced an anxiolytic-like effect after injection into the basolateral amygdala nuclei or the CA1 region of the hippocampus in the conflict drinking Vogel test in rats. The obtained results indicate that (S)-3,4-DCPG (10, 50 and 100 nmol/rat) produces no anticonflict effect in rats. We conclude that selective stimulation of mGlu8 receptors (a subtype of group III mGluRs) does not evoke anxiolytic-like activity, and that the mGlu8 receptors are of no significance for anxiolytic-like effects of group III mGluR agonists. PMID- 16382207 TI - Lack of effect of some dopamine and non-dopamine receptor ligands on amphetamine induced changes in the rat brain neuropeptide Y system. AB - None of the receptor ligands studied: the dopamine D2 antagonist eticlopride, D2/D3 antagonist haloperidol, alpha1-adrenergic antagonist prazosin, N-methyl-D aspartate antagonist MK-801, 5-HT2/D2 antagonist mianserin, D2/D3 agonist quinpirole and alpha2-adrenergic agonist clonidine co-administrated with repeated amphetamine (AMPH) injections blocked the AMPH-induced decrease in the striatal and nucleus accumbens neuropeptide Y (NPY) levels. Only the D1/D5 receptor antagonist SCH 23390 insignificantly attenuated this effect what suggests that these dopamine receptor subtypes may partially mediate AMPH effect on NPY. Moreover, the results of multiple administrations of the receptor ligands to control rats indicate that the NPY systems in both structures under study are modulated in the same manner by dopaminergic activity and differentially by adrenergic, N-methyl-D-aspartate and serotonergic activity. PMID- 16382209 TI - Application of magnetic resonance diffusion anisotropy imaging for the assessment neuroprotecting effects of MPEP, a selective mGluR5 antagonist, on the rat spinal cord injury in vivo. AB - Magnetic resonance diffusion anisotropy imaging (DAI) of the rat spinal cord after contusion using weight-drop method was used to study the neuroprotecting effect of 2-methyl-6-(phenylethynyl)-pyridine (MPEP), an mGluR5 receptor antagonist. Eighteen rats were used, divided into 3 groups of 6 animals: a reference group without any operation, a control group with injury and a test group with injury and MPEP. DAI was performed at 4.7 T at 1 h, 24 h, 48 h and 7 day after the injury. Locomotor function was evaluated using Basso, Beattie and Bresnahan (BBB) open field locomotor activity test each day starting one day after the injury. DAI results confirm positive effect of MPEP on the limitation of secondary excitotoxic injury in the spinal cord. PMID- 16382210 TI - Direct effects of neuroleptics on the activity of CYP2A in the liver of rats. AB - The aim of the present study was to investigate the influence of classic and atypical neuroleptics on the activity of rat CYP2A measured as a rate of testosterone 7alpha-hydroxylation. The reaction was studied in control liver microsomes in the presence of neuroleptics, as well as in microsomes of rats treated intraperitoneally (i.p.) for one day or two weeks (twice a day) with pharmacological doses (mg/kg) of the drugs (promazine, levomepromazine, thioridazine, perazine 10, chlorpromazine, haloperidol 0.3, risperidone 0.1, sertindole 0.05), in the absence of the neuroleptics in vitro. Most of the neuroleptics added in vitro to control liver microsomes decreased the activity of the rat CYP2A. Chlorpromazine (Ki = 11 microM) was the most potent inhibitor of the rat CYP2A among the studied drugs, whose effect was more pronounced than that of the other tested phenothiazines (Ki = 41-83 microM), haloperidol (Ki = 190 microM) or sertindole (Ki = 78 microM). Risperidone was not active in this respect. The investigated neuroleptics when given to rats in vivo for one day or two weeks--did not produce any indirect inhibitory effect on CYP2A via other mechanisms. The obtained results show direct inhibitory effects of phenothiazine neuroleptics on the activity of CYP2A in rat liver, which may be of physiological importance for the metabolism of testosterone, considering simultaneous inhibition of CYP2C11 and CYP3A by those drugs. PMID- 16382211 TI - Interactions between neuroleptics and CYP2C6 in rat liver--in vitro and ex vivo study. AB - The aim of the present study was to investigate the influence of classic and atypical neuroleptics on the activity of rat CYP2C6 measured as a rate of warfarin 7-hydroxylation. The reaction was studied in control liver microsomes in the presence of neuroleptics, as well as in microsomes of rats treated intraperitoneally for one day or two weeks (twice a day) with pharmacological doses (mg/kg) of the drugs (promazine, levomepromazine, thioridazine, perazine 10, chlorpromazine, haloperidol 0.3, risperidone 0.1, sertindole 0.05), in the absence of the neuroleptics in vitro. Some of the neuroleptics added in vitro to control liver microsomes decreased the activity of CYP2C6. Sertindole and levomepromazine (Ki = 25 and 31 microM, respectively) were the most potent inhibitors of the rat CYP2C6 among the drugs studied. Their effects were more pronounced than those of the other phenothiazines tested: thioridazine and chlorpromazine (Ki = 88 and 91 microM, respectively), promazine and perazine (Ki = 322 and 341 microM, respectively), risperidone (Ki = 414 microM) or haloperidol (Ki = 606 microM). The investigated neuroleptics--when given to rats in vivo for one day or two weeks--did not produce any indirect effect on CYP2C6 via other mechanisms, except for levomepromazine, which increased the activity of the enzyme after 24-h exposure. Therefore, the direct inhibitory effect of levomepromazine on CYP2C6 may be attenuated by an indirect mechanism at the beginning of the neuroleptic therapy. In summary, the obtained results show direct inhibitory effects of some phenothiazine neuroleptics and sertindole on the activity of CYP2C6 in vitro in rat liver microsomes. Considering relatively high pharmacological doses and therapeutic concentrations of phenothiazines, it seems that the inhibitory effect of levomepromazine (and other phenothiazines with Ki values below 100 microM) found in vitro may be of physiological and pharmacological importance in vivo. PMID- 16382212 TI - Plasma concentrations of adhesion molecules and chemokines in patients with essential hypertension. AB - Arterial hypertension vascular injury results in serious complications, such as left-ventricular hypertrophy and myocardial failure, ischemic heart disease and cerebral stroke. Currently, it is well known that inflammatory factors play a significant role in the mechanisms that trigger and enhance the remodeling of the vascular wall. A number of data suggest an important role of adhesion molecules and chemokines in this processes. The aim of this study was measuring the plasma levels of soluble Intercellular Adhesion Molecule 1 (sICAM-1) and Monocyte Chemoattractant Peptide1 (MCP-1) in patients with essential hypertension vs. healthy volunteers by ELISA method (R&D kits). sICAM-1 and MCP-1 levels were significantly higher in hypertensive patients compared to controls (sICAM-1: 279.2 +/- 8.8 ng/ml vs. 224.4 +/- 1.8 ng/ml; p < 0.001; MCP-1: 142.2 +/- 7 pg/ml vs. 95.4 +/- 36 pg/ml; p < 0.0001. Our results indicate that arterial hypertension alone (without inflammation, lipid and carbohydrate disorders) may increase the expression of these cytokines and contribute to the progression of endothelial injury. PMID- 16382213 TI - Multi-drug transporter MDR1 gene polymorphism and prognosis in adult acute lymphoblastic leukemia. AB - P-glycoprotein (P-gp), a membrane transporter encoded by MDR1 gene, influences pharmacokinetics of anti-cancer drugs and contributes to multi-drug resistance phenotype in adult acute lymphoblastic leukemia (ALL). In this study, we explored prognostic and functional role of single nucleotide polymorphism C3435T in MDR1 gene in 44 adult Caucasian patients with ALL. We found that the outcome of chemotherapy as well as MDR1 gene expression, P-gp expression and P-gp activity in isolated ALL blast cells were comparable among the patients carrying different MDR1 genotypes. Our results suggest that C3435T polymorphism in MDR1 gene is not a major prognosticator in adult ALL. PMID- 16382214 TI - Study on organic nitrates, part VIII. Pharmacological activity and nitric oxide generation capacity of nitrate derivatives of piperazine. AB - Organic nitrates, derivatives of piperazine, incubated with L-cysteine hydrochloride in phosphate buffer at appropriately high pH and temperature underwent decomposition, releasing nitric oxide (NO). NO generated in this reaction was quantitated by polarographic method using a selective ISO NO sensor. Spontaneously hypertensive rats were used in the tests of hypotensive activity of both investigated organic nitrates and their hydroxyl analogues. The experiments with both groups of compounds were performed under identical conditions. The results of the study demonstrated that the ability of individual compounds to release NO influenced in a different manner their pharmacological activity. Only the compound which released the largest quantity of NO during in vitro tests exhibited in vivo the most prolonged hypotensive effect and significant decrease in all arterial blood pressure parameters. PMID- 16382215 TI - Effect of 4-hydroxyandrost-4-ene-3,17-dione (formestane) on the bile secretion and metabolism of 4-(14)C-cholesterol to bile acids. AB - 4-Hydroxyandrost-4-ene-3,17-dione (formestane) is a selective aromatase inhibitor. It is indicated for postmenopausal patients with advanced breast cancer. The aim of the present study was to investigate the effect of 4 hydroxyandrost-4-ene-3,17-dione on the bile secretion and metabolism of 4-(14)C cholesterol to bile acid. The experiments were carried out in the ovariectomized and sham-operated female Wistar rats. Formestane (20 mg/kg, i.m., daily) was administered to animals for 2 weeks. Twenty four hours after the last drug administration, rats were anesthetized with ethyl urethane. 4-(14)C-cholesterol (740 kBq/kg, s.a. 2.28 GBq/mmol) was infused for 1 min by catheter inserted into the jugular vein. Bile samples were assayed for total 14C radioactivity 14C-bile acids were determined in bile (after thin-layer chromatographic separation) by the use of isotopic technique with liquid scintillator. Previous studies showed that systemic adverse effects occurred in about 12% of patients following intramuscular drug administration. Many of them such as hot flushes, vaginal spotting and emotional lability were related to the mechanism of action of formestane i.e. estrogen suppression. Lethargy, rash, nausea, dizziness, indigestion, ataxia, cramps and facial swelling have also been reported. The results of the present study have shown that formestane administered to the female ovariectomized rats decreased the bile secretion and diminished conversion of 4-(14)C-cholesterol to trihydroxy bile acids. The decreased synthesis of trihydroxy bile acids and increased concentrations of cholesterol and litocholic acid in bile may be associated with increased risk of gallstone formation. PMID- 16382216 TI - Guidelines for preventing the transmission of Mycobacterium tuberculosis in health-care settings, 2005. AB - In 1994, CDC published the Guidelines for Preventing the Transmission of Mycobacterium tuberculosis in HealthCare Facilities, 1994. The guidelines were issued in response to 1) a resurgence of tuberculosis (TB) disease that occurred in the United States in the mid-1980s and early 1990s, 2) the documentation of several high-profile health-care--associated (previously termed "nosocomial") outbreaks related to an increase in the prevalence of TB disease and human immunodeficiency virus (HIV) coinfection, 3) lapses in infection control practices, 4) delays in the diagnosis and treatment of persons with infectious TB disease, and 5) the appearance and transmission of multidrug-resistant (MDR) TB strains. The 1994 guidelines, which followed statements issued in 1982 and 1990, presented recommendations for TB infection control based on a risk assessment process that classified health-care facilities according to categories of TB risk, with a corresponding series of administrative, environmental, and respiratory protection control measures. The TB infection control measures recommended by CDC in 1994 were implemented widely in health-care facilities in the United States. The result has been a decrease in the number of TB outbreaks in health-care settings reported to CDC and a reduction in health-care-associated transmission of Mycobacterium tuberculosis to patients and health-care workers (HCWs). Concurrent with this success, mobilization of the nation's TB control programs succeeded in reversing the upsurge in reported cases of TB disease, and case rates have declined in the subsequent 10 years. Findings indicate that although the 2004 TB rate was the lowest recorded in the United States since national reporting began in 1953, the declines in rates for 2003 (2.3%) and 2004 (3.2%) were the smallest since 1993. In addition, TB infection rates greater than the U.S. average continue to be reported in certain racial/ethnic populations. The threat of MDR TB is decreasing, and the transmission of M. tuberculosis in health-care settings continues to decrease because of implementation of infection control measures and reductions in community rates of TB. Given the changes in epidemiology and a request by the Advisory Council for the Elimination of Tuberculosis (ACET) for review and update of the 1994 TB infection control document, CDC has reassessed the TB infection control guidelines for health-care settings. This report updates TB control recommendations reflecting shifts in the epidemiology of TB, advances in scientific understanding, and changes in health care practice that have occurred in the United States during the preceding decade. In the context of diminished risk for health-care-associated transmission of M. tuberculosis, this document places emphasis on actions to maintain momentum and expertise needed to avert another TB resurgence and to eliminate the lingering threat to HCWs, which is mainly from patients or others with unsuspected and undiagnosed infectious TB disease. CDC prepared the current guidelines in consultation with experts in TB, infection control, environmental control, respiratory protection, and occupational health. The new guidelines have been expanded to address a broader concept; health-care--associated settings go beyond the previously defined facilities. The term "health-care setting" includes many types, such as inpatient settings, outpatient settings, TB clinics, settings in correctional facilities in which health care is delivered, settings in which home-based health-care and emergency medical services are provided, and laboratories handling clinical specimens that might contain M. tuberculosis. The term "setting" has been chosen over the term "facility," used in the previous guidelines, to broaden the potential places for which these guidelines apply. PMID- 16382225 TI - Should you count your mice before they're weaned? Pool the pups. PMID- 16382226 TI - Should you count your mice before they're weaned? PMID- 16382228 TI - Should you count your mice before they're weaned? Silence not equal compliance. PMID- 16382229 TI - Should you count your mice before they're weaned? Start counting. PMID- 16382230 TI - Cachexia and sialorrhea in a female rat. PMID- 16382232 TI - Employee motivation in laboratory animal science: creating the conditions for a happy and productive staff. AB - High rates of employee turnover are the source of a considerable loss of time and resources, but managers are not always aware of the reasons that motivate employees to stay in their positions. The author compares prominent theories of employee motivation and then puts them to the test by surveying 82 cagewashers, animal caretakers, animal technicians, and supervisors working in a laboratory animal facility to determine the job characteristics that motivate them. PMID- 16382233 TI - The Study Support Associate: an alternative to the traditional animal care position. AB - The authors describe the development of a position in animal care that combines husbandry, veterinary care, breeding colony management, and in vivo research support. The integration of these tasks gives participating animal care staff more diverse responsibilities and leads to more effective communication and collaboration between researchers and husbandry personnel. PMID- 16382234 TI - Motivation of animal care technicians through recognition. AB - Keeping employees motivated is a challenge faced by managers in the field of laboratory animal science and in the business world at large. Using Maslow's 'Hierarchy of Needs' theory as a guide, the authors describe how managers can create a recognition program to keep employees feeling happy and rewarded. They discuss programs used at Bristol-Myers Squibb and share lessons learned from various programs. PMID- 16382235 TI - Close-up on lab animal microsurgery. PMID- 16382236 TI - Restoring function in exhausted CD8 T cells during chronic viral infection. AB - Functional impairment of antigen-specific T cells is a defining characteristic of many chronic infections, but the underlying mechanisms of T-cell dysfunction are not well understood. To address this question, we analysed genes expressed in functionally impaired virus-specific CD8 T cells present in mice chronically infected with lymphocytic choriomeningitis virus (LCMV), and compared these with the gene profile of functional memory CD8 T cells. Here we report that PD-1 (programmed death 1; also known as Pdcd1) was selectively upregulated by the exhausted T cells, and that in vivo administration of antibodies that blocked the interaction of this inhibitory receptor with its ligand, PD-L1 (also known as B7 H1), enhanced T-cell responses. Notably, we found that even in persistently infected mice that were lacking CD4 T-cell help, blockade of the PD-1/PD-L1 inhibitory pathway had a beneficial effect on the 'helpless' CD8 T cells, restoring their ability to undergo proliferation, secrete cytokines, kill infected cells and decrease viral load. Blockade of the CTLA-4 (cytotoxic T lymphocyte-associated protein 4) inhibitory pathway had no effect on either T cell function or viral control. These studies identify a specific mechanism of T cell exhaustion and define a potentially effective immunological strategy for the treatment of chronic viral infections. PMID- 16382237 TI - A var gene promoter controls allelic exclusion of virulence genes in Plasmodium falciparum malaria. AB - Mono-allelic expression of gene families is used by many organisms to mediate phenotypic variation of surface proteins. In the apicomplexan parasite Plasmodium falciparum, responsible for the severe form of malaria in humans, this is exemplified by antigenic variation of the highly polymorphic P. falciparum erythrocyte membrane protein 1 (PfEMP1). PfEMP1, encoded by the 60-member var gene family, represents a major virulence factor due to its central role in immune evasion and intravascular parasite sequestration. Mutually exclusive expression of PfEMP1 is controlled by epigenetic mechanisms involving chromatin modification and perinuclear var locus repositioning. Here we show that a var promoter mediates the nucleation and spreading of stably inherited silenced chromatin. Transcriptional activation of this promoter occurs at the nuclear periphery in association with chromosome-end clusters. Additionally, the var promoter sequence is sufficient to infiltrate a transgene into the allelic exclusion programme of var gene expression, as transcriptional activation of this transgene results in silencing of endogenous var gene transcription. These results show that a var promoter is sufficient for epigenetic silencing and mono allelic transcription of this virulence gene family, and are fundamental for our understanding of antigenic variation in P. falciparum. Furthermore, the PfEMP1 knockdown parasites obtained in this study will be important tools to increase our understanding of P. falciparum-mediated virulence and immune evasion. PMID- 16382238 TI - A lever-arm rotation drives motility of the minus-end-directed kinesin Ncd. AB - Kinesins are microtubule-based motor proteins that power intracellular transport. Most kinesin motors, exemplified by Kinesin-1, move towards the microtubule plus end, and the structural changes that govern this directional preference have been described. By contrast, the nature and timing of the structural changes underlying the minus-end-directed motility of Kinesin-14 motors (such as Drosophila Ncd) are less well understood. Using cryo-electron microscopy, here we demonstrate that a coiled-coil mechanical element of microtubule-bound Ncd rotates approximately 70 degrees towards the minus end upon ATP binding. Extending or shortening this coiled coil increases or decreases velocity, respectively, without affecting ATPase activity. An unusual Ncd mutant that lacks directional preference shows unstable nucleotide-dependent conformations of its coiled coil, underscoring the role of this mechanical element in motility. These results show that the force-producing conformational change in Ncd occurs on ATP binding, as in other kinesins, but involves the swing of a lever-arm mechanical element similar to that described for myosins. PMID- 16382239 TI - Global tests of biodiversity concordance and the importance of endemism. AB - Understanding patterns of biodiversity distribution is essential to conservation strategies, but severe data constraints make surrogate measures necessary. For this reason, many studies have tested the performance of terrestrial vertebrates as surrogates for overall species diversity, but these tests have typically been limited to a single taxon or region. Here we show that global patterns of richness are highly correlated among amphibians, reptiles, birds and mammals, as are endemism patterns. Furthermore, we demonstrate that although the correlation between global richness and endemism is low, aggregate regions selected for high levels of endemism capture significantly more species than expected by chance. Although areas high in endemism have long been targeted for the protection of narrow-ranging species, our findings provide evidence that endemism is also a useful surrogate for the conservation of all terrestrial vertebrates. PMID- 16382240 TI - Dissociable neural mechanisms supporting visual short-term memory for objects. AB - Using visual information to guide behaviour requires storage in a temporary buffer, known as visual short-term memory (VSTM), that sustains attended information across saccades and other visual interruptions. There is growing debate on whether VSTM capacity is limited to a fixed number of objects or whether it is variable. Here we report four experiments using functional magnetic resonance imaging that resolve this controversy by dissociating the representation capacities of the parietal and occipital cortices. Whereas representations in the inferior intra-parietal sulcus (IPS) are fixed to about four objects at different spatial locations regardless of object complexity, those in the superior IPS and the lateral occipital complex are variable, tracking the number of objects held in VSTM, and representing fewer than four objects as their complexity increases. These neural response patterns were observed during both VSTM encoding and maintenance. Thus, multiple systems act together to support VSTM: whereas the inferior IPS maintains spatial attention over a fixed number of objects at different spatial locations, the superior IPS and the lateral occipital complex encode and maintain a variable subset of the attended objects, depending on their complexity. VSTM capacity is therefore determined both by a fixed number of objects and by object complexity. PMID- 16382241 TI - Stem cell engraftment at the endosteal niche is specified by the calcium-sensing receptor. AB - During mammalian ontogeny, haematopoietic stem cells (HSCs) translocate from the fetal liver to the bone marrow, where haematopoiesis occurs throughout adulthood. Unique features of bone that contribute to a microenvironmental niche for stem cells might include the known high concentration of calcium ions at the HSC enriched endosteal surface. Cells respond to extracellular ionic calcium concentrations through the seven-transmembrane-spanning calcium-sensing receptor (CaR), which we identified as being expressed on HSCs. Here we show that, through the CaR, the simple ionic mineral content of the niche may dictate the preferential localization of adult mammalian haematopoiesis in bone. Antenatal mice deficient in CaR had primitive haematopoietic cells in the circulation and spleen, whereas few were found in bone marrow. CaR-/- HSCs from fetal liver were normal in number, in proliferative and differentiative function, and in migration and homing to the bone marrow. Yet they were highly defective in localizing anatomically to the endosteal niche, behaviour that correlated with defective adhesion to the extracellular matrix protein, collagen I. CaR has a function in retaining HSCs in close physical proximity to the endosteal surface and the regulatory niche components associated with it. PMID- 16382242 TI - Infectious disease eradication: poliomyelitis as a lesson in why "close" is not good enough. PMID- 16382243 TI - Locally advanced and metastatic breast cancer in a tertiary hospital. AB - INTRODUCTION: The breast cancer incidence among Singapore women has risen through the years and is now the highest in Asia. Despite efforts to promote a greater awareness of breast cancer among the public, a significant number of patients still present with locally advanced or metastatic breast cancer. Our study aims to evaluate the clinical and pathological characteristics between patients presenting with locally advanced (LABC) and metastatic breast cancer (MBC) and those presenting with early breast cancer (EBC), to identify factors that predict for advanced disease. MATERIALS AND METHODS: We reviewed 622 patients who were newly diagnosed with invasive breast cancer in our department over a 4-year period from January 2000 to December 2003. Patient and tumour characteristics including age, parity, family history, tumour size and histology, grade and hormonal receptor status were analysed. Comparisons were made between those with EBC and those with LABC and MBC, as well as between Malay women and women of other ethnic groups. RESULTS: One hundred and thirty-four patients (21.5%) presented with either LABC or MBC. Adjusted analysis found that these patients were older and more likely to be nulliparous than those with EBC. Older patients tend to have larger tumours, but otherwise, age and parity did not correlate with tumour histology, grade or hormonal status. It was noted that Malay women, who were more likely to present with LABC or MBC, were more likely to have oestrogen receptor- and progesterone receptor-negative tumours. CONCLUSIONS: Older women and those who were nulliparous were found more likely to present with LABC and MBC. However, age and parity did not appear to be related to tumour histology, grade and hormonal status. Given that tumour size and stage have the greatest impact on overall survival, efforts to raise public awareness of the benefits of early detection and treatment should be continued, and possibly directed towards these groups of women who appear to be at an increased risk of presenting late. PMID- 16382244 TI - Clinical features associated with nasal Staphylococcus aureus colonisation in Chinese children with moderate-to-severe atopic dermatitis. AB - INTRODUCTION: Staphylococcus aureus colonisation/infection is common in children with atopic dermatitis (AD). MATERIALS AND METHODS: We evaluated the nasal and body swabs of Chinese children with moderate-to-severe AD as assessed using SCORing-Atopic-Dermatitis (SCORAD) score. Swabs were taken from the right nostril, 5 flexural sites (anterior neck, antecubital fossae and popliteal fossae) and the skin area most severely affected (with oozing/crusting) for bacteriologic culture. RESULTS: Fifty-five children (30 males and 25 females) were evaluated. Moderate-to-heavy growth of S. aureus was present in 12 (22%) of the nasal swabs, and in 1 or more flexural swabs of 32 (58%) of these children. Only 7 (35%) out of the 20 patients who had swabs taken from the worst skin area had moderate-to-heavy growth of S. aureus. Significant nasal S. aureus colonisation was associated with higher total (P=0.029) and objective SCORAD scores (P=0.040), more extensive disease (P=0.025), the presence of oozing or crusting (P=0.023) and higher eosinophil counts (P=0.038). All specimens of methicillin-sensitive S. aureus were sensitive to cloxacillin and 71% to erythromycin. Methicillin-resistant S. aureus (MRSA), sensitive to vancomycin, was only isolated in 1 patient. CONCLUSIONS: In this study, S. aureus is a principal pathogen. Cloxacillin and first-generation cephalosporins have a favourable sensitivity profile even in children with moderate and severe atopic dermatitis. The anterior nares are an important harbour for S. aureus and significant nasal S. aureus colonisation was clinically associated with more extensive lesions and the presence of oozing or crusting. PMID- 16382245 TI - Evaluation of surgical and anaesthesia response times for crash caesarean sections--an audit of a Singapore hospital. AB - INTRODUCTION: The Royal College of Obstetricians and Gynaecologists published the "Organisational Standards for Maternity Services" in 1995, in which they proposed that there be a maximum decision-to-delivery time of 30 minutes for urgent caesarean sections (CS). In 1997, our institution established a protocol for extremely urgent ("crash") CS to expedite delivery time and to conform to this standard. MATERIALS AND METHODS: The objective of this prospective audit was to determine the surgical and anaesthesia response times in our institution after the protocol had been implemented. The audit was conducted in KK Women's and Children's Hospital from February 2003 to January 2004, over a 12-month period. Upon activation of a "crash" CS, the attending anaesthetist was required to record the decision-to-anaesthesia time, decision-to-delivery time and the perinatal outcome. RESULTS: Ninety-eight cases of "crash" CS were identified from a total of 3629 elective and non-elective CS, with 80 cases having complete data. The mean decision-to-delivery interval was 7.7 min+/-3.0 (SD) with 100% of deliveries made within 17 minutes. The mean decision-to-anaesthesia time was 3.5 min+/-2.0 (SD) with all the patients anaesthetised within 10 minutes. The majority (88.8%) of the patients had general anaesthesia for "crash" CS while the rest had successful epidural block extension. There was no significant difference in the decision-to-delivery interval or mean cord blood pH with respect to the type of anaesthesia given. CONCLUSIONS: We achieved 100% deliveries within the proposed 30-minute decision-to-delivery time interval by implementing a protocol for "crash" CS. Both general anaesthesia and extension of existing epidural block are acceptable modes of anaesthesia and do not delay delivery of the fetus. PMID- 16382246 TI - Carbon monoxide poisoning: a five year review at Tan Tock Seng Hospital, Singapore. AB - INTRODUCTION: Carbon monoxide poisoning (COP) is one of the leading causes of death from poisoning worldwide. There is no published study of COP in Singapore so far. MATERIALS AND METHODS: All patients admitted with the diagnosis of COP to Tan Tock Seng Hospital (TTSH) over 5 years from 1999 to 2003 were retrospectively reviewed. The diagnosis was based on a history of potential exposure to carbon monoxide (CO) and elevated levels of carboxyhaemoglobin (COHb). The causes, demographic data, clinical presentations, management and complications were analysed. RESULTS: There were 12 patients with COP. Their average age was 38.9 (+/-11.8) years, with a male-to-female ratio of 3:1. Accidental COP (58.3%) was more common than intentional COP (41.7%). The most common cause of accidental COP was smoke inhalation from a faulty vehicle. Gas stove was the most preferred source for intentional poisoning. Presenting features were headache (83.3%), confusion (83.3%), coma (12.7%) and agitation (8.3%). The mean COHb level on admission was 35.9% (+/-13.6). All were treated with 100% oxygen. All the patients achieved normal levels of COHb within 24 hours of admission. Two (16.7%) required intubation for airway protection as they were comatose on arrival, of which 1 presented with very high level of COHb (48.1%) and was the only patient to be treated with hyperbaric oxygen. Acute complications were globus pallidus infarction (16.6%), acute respiratory distress syndrome (8.3%) and myocardial ischaemia (8.3%). Most of the patients (91.7%) were discharged well from the hospital. One patient developed parkinsonism after a follow-up of 2 years. There were no deaths. CONCLUSION: COP is relatively uncommon in Singapore. It has a low rate of short- and long-term complications. PMID- 16382247 TI - Plasma filtration. AB - Therapeutic plasma exchange (TPE) or plasmapheresis involves the separation of plasma from whole blood. In so doing, plasma-borne humoral disease mediators are removed from the body. This can attenuate the course and severity of the underlying disease. Diseases that can be treated with TPE are classified into the following categories: (1) endocrinological, (2) neurological, (3) renal/rheumatological, and (4) haematological. TPE is adjuvant in most of these settings. Disease-specific pharmacological treatment remains the cornerstone of treatment in many of these conditions. Plasma separation can be achieved with either (1) centrifugation (CF) or (2) membrane plasma filtration (PF). The latter is the focus of this review. It can be performed using either a continuous renal replacement therapy (CRRT) or haemodialysis (HD) machine. Standard plasma filtration has also been modified to incorporate sorbent technology which obviates the need for plasma volume replacement fluids. Larger clinical issues such as timing of initiation and intensity of therapy are examined. PMID- 16382248 TI - Tsunami in South Asia: what is the risk of post-disaster infectious disease outbreaks? AB - The World Health Organization has warned that in the aftermath of the recent tsunami, infectious disease outbreaks will add to the heavy toll of the disaster itself, possibly even doubling the number of casualties. However, many experts believe the risks of infectious disease outbreaks following natural disasters have been overemphasised and have led to unnecessary and potentially harmful public health activities. This paper discusses the risk and prevention strategies of potential infectious diseases in the aftermath of the tsunami based on a literature review of previous similar disasters and current evidence. Infectious disease outbreaks, if any, will most likely be the consequence of post-tsunami camp situations involving large displaced populations rather than the tidal wave itself. Lessons have been learned from previous large-scale humanitarian crises about the provision of aid and the mitigation of epidemics. This paper examines the risk and preventive strategies of vector- and food/water-borne diseases, measles, acute respiratory infections and meningitis. Alert thresholds at which to trigger outbreak investigations, and standardised guidelines with regard to their control are outlined, based on the Sphere Project. PMID- 16382249 TI - Inflammatory tumour of the retroperitoneum--a case report. AB - INTRODUCTION: Neoplastic growths of myofibroblasts occurring on a background of plasma cell and lymphocytic proliferation have been designated as inflammatory myofibroblastic tumours (IMTs). These unusual tumours were first described in pulmonary location in adults. Though extrapulmonary masses have been reported in children; retroperitoneal growths are exceedingly rare. We report a case of retroperitoneal IMT that presented with constitutional symptoms without any palpable abdominal mass. CLINICAL PICTURE: A previously well 12-year-old boy presented with fever, right-sided flank pain and weight loss of 1-month duration. There were no foci of infection. The erythrocyte sedimentation rate (ESR) was raised; the white cell count was normal. An abdominal computed tomography (CT) scan revealed a right suprarenal tumour measuring 3.5 cm without any calcification. The urinary catecholamines and vanilmandelic acid were normal. TREATMENT: A laparotomy with complete excision of the tumour was performed. Final histology revealed an inflammatory myofibroblastic tumour without any correlates of aggressive behaviour. OUTCOME: Postoperatively, the constitutional symptoms of fever, weight loss and raised ESR normalised. Follow-up CT was normal and further treatment was not necessary. CONCLUSION: Although rare, IMTs should be considered in any abdominal solid tumour with associated constitutional and laboratory features of an inflammatory response. Complete surgical excision is effective treatment for biologically benign tumours. PMID- 16382250 TI - Periampullary diverticulum: a case of bleeding from a periampullary diverticulum. AB - INTRODUCTION: Duodenal diverticula are uncommon and usually asymptomatic. Complications like bleeding, perforation and biliary fistulae are rare. The management of a bleeding duodenal diverticulum can be challenging. With the improvement of endoscopic techniques, many of the cases reported in the literature were managed with endoscopic methods. We present a case report of bleeding duodenal diverticulum. CLINICAL FEATURES: The patient was treated successfully with endoscopic haemostasis during her first epidsode when she initially presented with bleeding duodenal diverticulum, but recurred after 2 months. TREATMENT: Despite initial endoscopic haemostasis during her second episode, she rebled after 2 days, necessitating surgical management. OUTCOME: After suture ligation of the ulcer, the patient recovered and there was no more recurrence. CONCLUSION: Periampullary diverticulum is a rare source of gastrointestinal bleeding, which can be challenging to diagnose and treat. A multidisciplinary approach encompassing radiology, endoscopy and surgery is most effective. PMID- 16382251 TI - Dural sinus thrombosis after minor head injury in a child. AB - INTRODUCTION: Dural sinus thrombosis following minor head injury is rare. We report such a case in a child after mild head injury. CLINICAL PICTURE: A 4-year old child presented with giddiness and vomiting after a fall. Clinical examination was unremarkable. Magnetic resonance venogram revealed thrombosis of the right sigmoid and transverse sinuses. TREATMENT: The patient was managed conservatively. OUTCOME: Repeat scans 10 weeks after injury showed recanalisation of the thrombosis. CONCLUSION: Dural sinus thrombosis should be excluded in children presenting with persistent giddiness and vomiting after minor head injury. PMID- 16382252 TI - Beyond Ortner's syndrome--unusual pulmonary complications of the giant left atrium. AB - INTRODUCTION: The giant left atrium (GLA) is a complication of severe mitral valve disease and causes morbidity by compressing adjacent intra-thoracic structures. CLINICAL PICTURE: We report 2 cases of unusual pulmonary complications of the GLA. Case 1 developed recurrent collapse of the left lung due to left main bronchus compression. Case 2 was diagnosed with right middle lobe compression and collapse. TREATMENT AND OUTCOME: Case 1 was successfully treated by mitral valve replacement and left atrial reduction surgery. Case 2 was treated conservatively. CONCLUSION: Pulmonary atelectasis may occur in patients with GLA due to bronchopulmonary compression. Surgical management with valve replacement and atrial reduction may be necessary to relieve airway compression. PMID- 16382253 TI - Fetus-in-fetu in the pelvis: report of a case and literature review. AB - INTRODUCTION: Fetus-in-fetu is an extremely rare condition in which a malformed fetus is found in the body of its twin. To our knowledge, fewer than 100 cases have been reported. Wide variations of presentation have been described, although its embryo-pathogenesis and differentiation from a teratoma have not been well established. CLINICAL PICTURE: We describe a male neonate with a fetoid-like mass in his pelvis associated with bilateral undescended testes. The mass was detected on prenatal ultrasound scans. The diagnosis of fetus-in-fetu was considered prenatally and confirmed on a computed tomography scan after birth. OUTCOME: The mass was successfully excised. Histological examination, accompanied by a review of the literature, confirmed that the mass had features consistent with a fetus in-fetu. CONCLUSIONS: Although an extremely rare clinical entity, fetus-in-fetu can be diagnosed prior to surgery with current imaging modalities. When it arises in the retroperitoneum of a male infant, it can hinder the descent of the testes. Complete excision is curative. PMID- 16382254 TI - New treatment for atopic dermatitis--facts, comparisons and uncertainties. PMID- 16382256 TI - Oesophageal carcinoma: patient's refusal for surgery may be the right choice. PMID- 16382257 TI - Bivalirudin vs heparin in percutaneous coronary intervention: a pooled analysis. AB - OBJECTIVE: This study evaluates outcomes with bivalirudin vs heparin in various patient subgroups and the overall population during percutaneous coronary interventions (PCI). BACKGROUND: Recent data suggest that bivalirudin, a reversible direct thrombin inhibitor, provides ischemic protection superior to heparin and comparable to heparin plus glycoprotein (GP) IIb/IIIa inhibitors but with significantly fewer bleeding complications. Whether this advantage persists in different subgroups has not been fully defined. To our knowledge, this is the largest pooled analysis of bivalirudin to date. METHODS: Four randomized controlled trials were identified that compared bivalirudin to heparin (with or without GP IIb/IIIa inhibitors) in PCI. The incidence of death, myocardial infarction (MI), revascularization, and major bleeding at 48 hours was compared between these two agents overall and in patients with and without diabetes mellitus, hypertension, renal insufficiency, and advanced age. RESULTS: The trials consisted of 11,638 patients (bivalirudin, 5,861; heparin, 5,777). There were no significant differences in patient characteristics between the two groups. At 48 hours, the incidence of death, MI, revascularization, and major bleeding was significantly reduced in the bivalirudin group (7.8% vs 1.08%, P < .001); individual ischemic end points were significantly reduced for death (0.01% vs 0.02%, P = .049) and revascularization (2.0% vs 2.7%, P = .02), with similar reductions for major bleeding (2.7% vs 5.8%, P < .001). Subgroup analysis was generally consistent with the overall findings. CONCLUSION: This analysis further supports the superiority of bivalirudin compared with heparin. Bivalirudin provides excellent ischemic protection with a significant reduction of bleeding complications, even in high-risk subgroups. PMID- 16382258 TI - Metabolic modulators for chronic cardiac ischemia. AB - Many patients with ischemic heart disease continue to experience anginal symptoms despite revascularization and treatment with antianginal medications. The effectiveness of current anti-ischemic medications is limited by their hemodynamic side effects, such as hypotension and bradycardia, which result in compromised organ perfusion. In this article, we review five novel agents (ranolazine, trimetazidine, L-carnitine, ribose, and dichloroacetate) under investigation for treatment of ischemic heart disease that work by enhancing the efficiency of the myocardium, rather than decreasing its work. This new paradigm promises to eliminate these side effects. PMID- 16382259 TI - Intramyocardial injection of allogenic bone marrow-derived mesenchymal stem cells without immunosuppression preserves cardiac function in a porcine model of myocardial infarction. AB - BACKGROUND: We investigated the efficacy of directly injected allogenic bone marrow-derived mesenchymal stem cells in improving left ventricular function in a porcine model of myocardial infarction. METHODS: Left ventricular infarction was created in 16 adult Yorkshire pigs by coil embolization and thrombotic occlusion distal to the second diagonal artery. One month after myocardial infarction was induced, the animals were randomized to either direct injection of allogenic mesenchymal stem cells or sham treatment (culture medium). Allogenic bromodeoxyuridine-labeled mesenchymal stem cells (2 +/- 0.1 x 10(8)) were directly injected into the infarct and peri-infarct areas during an open chest procedure. No immunosuppressive therapy was used. The left ventricular function was measured using serial biplane left ventricular angiography at baseline, 30, 60, and 90 days before sacrifice. Mesenchymal stem cells were localized using bromodeoxyuridine, and differentiation of mesenchymal stem cells was assessed by confocal microscopic colocalization of bromodeoxyuridine with immunofluorescent antibodies specific for cardiomyocytes (troponin I and MF-20) and endothelial cells (von Willebrand factor). RESULTS: Mesenchymal stem cells labeled with bromodeoxyuridine engrafted the peri-infarct zone and colocalized with both cardiomyocyte-specific and endothelial cell-specific immunofluorescence. No intramyocardial bromodeoxyuridine was observed in sham-treated animals. At the time of the mesenchymal stem cell injection 30 days after myocardial infarction, the left ventricular ejection fraction (LVEF) was 58% +/- 3% in mesenchymal stem cell-treated pigs and 56% +/- 2% in sham-treated pigs (P = NS). LVEF deteriorated progressively thereafter in untreated pigs (8.5% and 10.5% decline at 60 days and 90 days after myocardial infarction, respectively), but was preserved in mesenchymal stem cell-treated pigs (2.1% increase and -2.0% decline at 60 and 90 days post-MI respectively) (P < .05). CONCLUSIONS: Direct intramyocardial injection of mesenchymal stem cells results in successful intramyocardial engraftment and differentiation into cardiomyocytes and endothelial cells and preserves left ventricular function after myocardial infarction in pigs. PMID- 16382260 TI - Sarcoplasmic reticulum adenosine triphosphatase overexpression in the L-type Ca2+ channel mouse results in cardiomyopathy and Ca2+ -induced arrhythmogenesis. AB - BACKGROUND: Overexpression of the L-type voltage-dependent calcium channel alpha(1C)-subunit (L-VDCC OE) in transgenic mice results in adaptive hypertrophy followed by a maladaptive phase associated with a decrease in sarcoplasmic reticulum adenosine triphosphatase (SERCA)2a expression at 8 to 10 months of age. Overexpressing SERCA to manipulate calcium (Ca(2+)) cycling and prevent pathologic phenotypes in some models of heart failure has been proven to be a promising genetic strategy. OBJECTIVE: In this study we investigated whether genetic manipulation that increases Ca(2+) uptake into the sarcoplasmic reticulum by overexpressing SERCA1a (skeletal muscle specific) into the L-VDCC OE background could restore or further deteriorate Ca(2+) cycling, contractile dysfunction, and electrical remodeling in the heart failure phenotype. RESULTS: We found that the survival rate of L-VDCC OE/SERCA1a OE double transgenic mice decreased by 50%. L-VDCC OE/SERCA1a OE mice displayed an accelerated phenotype of severe dilation of both ventricles associated with deteriorated left ventricular function. Voltage clamp experiments revealed enhanced increased inward Ca(2+) current density and decreased the transient outward potassium current. Action potential duration in double transgenic ventricular myocytes was prolonged, and isoproterenol induced early after depolarization. These mice demonstrated a high incidence of spontaneous left ventricular arrhythmia. Expression of the proarrhythmic signaling protein Ca(2+)/calmodulin-dependent kinase II (CaMKII) was increased while connexin43 expression was decreased, defining an important putative mechanism in the electrophysiologic disturbances and mortality. CONCLUSIONS: Despite previous reports of improved cardiac function in heart failure models after SERCA intervention, our results advocate the need to elucidate the involvement of augmented Ca(2+) cycling in arrhythmogenesis. PMID- 16382261 TI - A vigilant, hypoxia-regulated heme oxygenase-1 gene vector in the heart limits cardiac injury after ischemia-reperfusion in vivo. AB - OBJECTIVES: The effect of a cardiac specific, hypoxia-regulated, human heme oxygenase-1 (hHO-1) vector to provide cardioprotection from ischemia-reperfusion injury was assessed. BACKGROUND: When myocardial ischemia and reperfusion is asymptomatic, the damaging effects are cumulative and patients miss timely treatment. A gene therapy approach that expresses therapeutic genes only when ischemia is experienced is a desirable strategy. We have developed a cardiac specific, hypoxia-regulated gene therapy "vigilant vector'' system that amplifies cardioprotective gene expression. METHODS: Vigilant hHO-1 plasmids, LacZ plasmids, or saline (n = 40 per group) were injected into mouse heart 2 days in advance of ischemia-reperfusion injury. Animals were exposed to 60 minutes of ischemia followed by 24 hours of reperfusion. For that term (24 hours) effects, the protein levels of HO-1, inflammatory responses, apoptosis, and infarct size were determined. For long-term (3 week) effects, the left ventricular remodeling and recovery of cardiac function were assessed. RESULTS: Ischemia-reperfusion resulted in a timely overexpression of HO-1 protein. Infarct size at 24 hours after ischemia-reperfusion was significantly reduced in the HO-1-treated animals compared with the LacZ-treated group or saline-treated group (P < .001). The reduction of infarct size was accompanied by a decrease in lipid peroxidant activity, inflammatory cell infiltration, and proapoptotic protein level in ischemia-reperfusion-injured myocardium. The long-term study demonstrated that timely, hypoxia-induced HO-1 overexpression is beneficial in conserving cardiac function and attenuating left ventricle remodelling. CONCLUSIONS: The vigilant HO 1 vector provides a protective therapy in the heart for reducing cellular damage during ischemia-reperfusion injury and preserving heart function. PMID- 16382262 TI - Chlorthalidone improves endothelial-mediated vascular responses in hypertension complicated by nondiabetic metabolic syndrome. AB - BACKGROUND: The study was conducted to evaluate the vascular effects of chlorthalidone, a distal tubule-acting natriuretic agent, in hypertensive patients with nondiabetic metabolic syndrome, an insulin-resistant condition characterized by endothelial dysfunction and high risk for diabetes mellitus development. METHODS: Thirteen untreated hypertensive patients with Adult Treatment Panel-III-defined nondiabetic metabolic syndrome were assigned to 3 month treatment with chlorthalidone. The end-points were baseline and post treatment evaluation of (1) forearm blood flow (strain-gauge plethysmography) responses to graded intra-arterial acetylcholine infusion to test endothelial mediated vasomotor function, with sodium nitroprusside as a control for endothelium-independent vasodilatation; (2) minimum forearm vascular resistance, the ratio of mean blood pressure and maximal blood flow in response to 13-minute arterial occlusion, as a hemodynamic correlate of arteriolar structure; and (3) transcapillary albumin escape rate (the 1-hour decay rate of (125)I-albumin, 6-8 microC ev) as a measure of systemic capillary permeability. Additional measurements included baseline and posttreatment lipids, fasting, and postload glucose and insulin as well as the homeostasis model assessment, an index of insulin sensitivity. RESULTS: Chlorthalidone reduced blood pressure, augmented acetylcholine-mediated vasodilatation, decreased minimum forearm resistance, and slowed the transcapillary albumin escape rate. Metabolic parameters did not change significantly except for an increase in low-density lipoprotein cholesterol levels. CONCLUSIONS: Chlorthalidone improved endothelial function, reversed abnormal arteriolar structure, and slowed albumin permeation in hypertensive patients with nondiabetic metabolic syndrome. PMID- 16382263 TI - Heparin therapy leads to platelet activation and prolongation of PFA-100 closure time. AB - BACKGROUND: Heparin is used in the treatment of venous and arterial thromboembolic events, including unstable angina. Once unfractionated heparin is discontinued during the acute phase of unstable angina, it has been demonstrated that the disease process may be reactivated within hours. It is hypothesized that this reactivation may be a result of direct platelet activation by heparin that can linger even after the drug itself has been stopped. Prior studies have shown that heparin can either increase or decrease platelet activation. More recent studies have also shown conflicting effects of unfractionated heparin on PFA-100 testing. METHODS: We studied the in-vitro effects of unfractionated heparin on platelet function and PFA-100 testing. Unfractionated heparin was incubated with whole blood taken from 18 healthy volunteers. Platelet activation and aggregation was assessed with and without the presence of heparin. RESULTS: Platelet aggregation and activation were increased in the presence of heparin. Unfractionated heparin also significantly prolonged collagen/adenosine diphosphate closure time but did not affect collagen/epinephrine closure time. CONCLUSIONS: Unfractionated heparin leads to direct platelet activation and increases platelet aggregation in vitro. Unfractionated heparin causes prolongation of the collagen/adenosine diphosphate closure time in PFA-100 testing, possibly as a result of direct binding to von Willebrand factor in solution and interference with von Willebrand factor-glycoprotein Ib binding. PMID- 16382264 TI - Do all patients with coronary artery disease benefit from angiotensin converting enzyme inhibitors? AB - Cumulative evidence supports the use of angiotensin-converting enzyme (ACE) inhibitors for stable coronary artery disease in patients with and without heart failure. The dose and unique properties of ACE inhibitors, trial data, differences in trial design and demographics, may all contribute to variable responses in clinical outcomes. Pending direct comparator clinical trials between a tissue ACE inhibitor vs a plasma ACE inhibitor, evidence indicates that both ramipril and perindopril can be recommended for secondary risk prevention. PMID- 16382265 TI - A changing pattern of infection after major vascular reconstructions. AB - Wound and graft infection can occur in more than 40% of patients undergoing vascular reconstructions for peripheral arterial disease (PAD). A recent increase in the frequency and severity of infections, as well as a change in the microorganisms recovered, led us to undertake a retrospective case-controlled study of wound/graft infections at this institution. The medical records of all patients undergoing vascular reconstruction for PAD during the previous 36 months were reviewed. Patient demographics, graft location and conduit, infection location, causative microorganisms, and factors potentially associated with development of infection were recorded. Infections were classified according to a modification of the CDC criteria into superficial incisional, deep incisional, or involving the graft (body only, anastomosis without disruption, or anastomosis with disruption). Univariate and multivariate regression analyses were used to identify factors associated with the development of infection. Four hundred ten (84 aortic, 41 extraanatomic, and 285 infrainguinal) revascularization procedures were performed in 217 men and 193 women with a mean age of 62 years (range 43 88). The infection rate for the entire group was 11.0% (45/410). Eighty percent (36/45) occurred after infrainguinal reconstructions and 64% (29/45) of the infections involved the groin incision. Direct involvement of the graft occurred in 67% (30/45), and 27% (12/45) presented with anastomotic disruption. Of the infrainguinal infections, in situ and prosthetic reconstructions were associated with a significantly higher rate of infection than reversed vein grafts tunneled anatomically (p <0.001, chi-square analysis). Patients with nonautogenous grafts (24 expanded polytetrafluoroethylene and 2 bovine) presented with more advanced infections involving the graft (20/26 procedures) and were more likely to present with anastomotic disruption (11/26). Staphylococcus aureus was isolated in the majority of infections (64%) and in all cases involving graft disruption. Multivariate regression analysis identified the following factors associated with development of infection: previous hospitalization (p = 0.03), a younger age (p = 0.047), and the presence of a groin incision (p = 0.04). Twenty-five percent of graft infections resulted in major amputation, and 11% of patients with graft infection died as a result. The incidence, morbidity, and mortality of infections in vascular reconstructions for PAD are increasing dramatically, particularly in infrainguinal reconstructions involving groin incisions. Perioperative antibiotic selection should be modified to include coverage for all Staphylococcal subspecies and hospitalization before surgical procedures should be avoided. PMID- 16382266 TI - An overview of matrix metalloproteinases in the pathogenesis and treatment of abdominal aortic aneurysms. AB - Recent basic and clinical research has established a link between the pathogenesis of abdominal aortic aneurysms (AAA) and matrix metalloproteinases (MMP). The discovery of the influence of MMPs on in vitro and in vivo aneurysm development has yielded promising information that may eventually decode the pathogenetic factors affecting the initiation and growth rate of AAAs. In this review, an analysis of MMPs involved in AAA disease is presented, including the data from recent research studies and planned clinical drug trails designed to retard the AAA growth by inhibiting MMP activity. PMID- 16382267 TI - The effect of secondary operations on mortality following abdominal aortic aneurysm repair in the United States: 1988-2001. AB - Certain complications following open repair of abdominal aortic aneurysms (AAAs) require additional operations or invasive procedures. The purpose of this study was to determine the effect of secondary interventions on mortality rate following open repair of intact and ruptured AAAs in the United States. Clinical data on 98,193 patients treated from 1988 to 2001 with an International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) primary procedure code 38.44 (resection of the abdominal aorta with replacement) were analyzed. Demographic factors, types of secondary interventions, and in hospital mortality rates were assessed by univariate and multivariate logistic regression analysis (SPSS Version 11.0, Chicago, IL). The database utilized in this study was The Nationwide Inpatient Sample (NIS). The mortality rate was 4.5% in the intact AAA group and 45.5% in the ruptured AAA group. The rate of secondary operations and procedures was much higher in the ruptured AAA group, especially related to renal failure (5.52% vs 1.49%, p <0.001); respiratory failure (3.67% vs 0.71%, p <0.001); postoperative bleeding (2.41% vs 0.81%, p <0.001); or colonic ischemia (2.38% vs 0.36%, p <0.001). Increased mortality following open repair of intact AAAs accompanied: peripheral artery angioplasty/stenting (OR, 1.25; 95% CI, 1.04-1.51; p = 0.018); coronary artery angioplasty/stenting (OR, 1.68; 95% CI, 1.05-2.70; p = 0.031); inferior vena cava (IVC) filter placement (OR, 2.02; 95% CI, 01.31-3.1; p = 0.001); vascular reconstruction or thromboembolectomy (OR, 2.05; 95% CI, 1.9-2.22; p <0.001); lower extremity amputation (OR, 4.09; 95% CI, 2.78-6.0; p <0.001); coronary artery bypass (OR, 6.71; 95% CI, 3.74-12.03; p <0.001); operations for postoperative bleeding (OR, 6.92; 95% CI, 5.71-8.4; p <0.001); initiation of hemodialysis (OR, 10.52; 95% CI, 9.22-12.01; p <0.001); tracheostomy (OR, 11.9; 95% CI, 9.86-14.37; p <0.001); and colectomy (OR, 16.22; 95% CI, 12.55-20.95; p <0.001). Increased risk of mortality following open repair of ruptured AAAs accompanied the following: operations for postoperative bleeding (OR, 1.5; 95% CI, 1.22-1.85; p <0.001); colectomy (OR, 1.63; 95% CI, 1.32-2.01; p <0.001); and initiation of hemodialysis (OR, 2.66; 95% CI, 2.30-3.08; p <0.001). The only independent variable in this group associated with decreased risk of in-hospital mortality was IVC filter placement (OR, 0.41; 95% CI, 0.27-0.64; p <0.001). This study confirms the perception that additional operations or invasive procedures following open repair of AAA entail significantly worse in-hospital mortality rates, especially when related to colonic ischemia, respiratory failure, and renal failure. PMID- 16382268 TI - Deciphering mesenteric venous thrombosis: imaging and treatment. AB - The principal cause of a high mortality rate in mesenteric vein thrombosis (MVT) is a delay in diagnosis. Recent data indicate that the mortality rate is decreasing owing to earlier diagnosis and anticoagulation. The authors examined the treatment profile of MVT to see how the increased use of imaging and early anticoagulation has impacted this process. They retrospectively analyzed the treatment paradigm with acute MVT at one institution over a 10-year period. Twenty-three patients were identified. Data were analyzed using chi-squares and Student's t tests. Twenty-three patients (11 men and 12 women with an average age of 51.74 +/-14.8 years) were identified with acute MVT between the years of 1993 and 2003. Five patients had splenic vein thrombosis, 17 had superior mesenteric vein thrombosis, 1 had inferior mesenteric vein thrombosis, and 12 had portal vein thrombosis. Nine patients had combination mesenteric vein segment thrombosis. Thrombolytics were utilized in a total of 6 patients. Four of the 6 patients in whom lytics were utilized had combined mesenteric vein thrombosis; however, these 4 patients did not require surgical intervention. There was no significant difference in length of hospital stay between patients taking lytics versus patients treated with traditional anticoagulation with heparin (p = 0.291). A hypercoagulable state was identified in 66.7% of the patients. Four patients required surgical intervention. The overall mortality rate was 8.7% (2 of 23). The use of thrombolytics was associated with a significant mortality (p = 0.04). The use of antibiotics made no difference in mortality (p = 0.235), nor did antibiotic use influence length of hospitalization (p = 0.192). MVT is relatively rare, and often the delay in diagnosis increases the mortality rate. In the majority of cases prompt anticoagulation will preserve bowel viability and decrease mortality and morbidity rates. The majority of patients do not need surgery. There is a marked increase in mortality rate when these patients progress to surgical intervention. An increased awareness and early diagnosis has led to decreased morbidity and mortality rates. PMID- 16382269 TI - Comparison of CT and catheter arteriography for evaluation of peripheral arterial disease. AB - Computed tomographic arteriography (CTA) has emerged as a promising technique for less invasive imaging of the lower extremity arteries. The aim of this study was to determine the concordance between CTA and catheter arteriography (CA) in patients with peripheral arterial disease (PAD). Twenty-five patients underwent both CTA and CA, and each set of images was interpreted independently by 3 readers. The infrarenal arteries were divided into 16 segments, and each segment was scored as: 1 = stenosis <50%; 2 = 50-99% stenosis; 3 = occlusion. Modal scores from 3 readers were used to compare results for each segment, with CA assumed to represent true arterial anatomy. Agreement between CTA and CA readings was defined as: concordance (modal scores were identical); moderate discrepancy (MD) (modal scores differed by 1); or severe discrepancy (SD) (modal scores differed by 2). In total, 718 segments were assessed by both CTA and CA. For all segments, the sensitivity and specificity of CTA for <50% stenosis was 86% and 90%; for 50-99% stenosis, sensitivity and specificity were 79% and 89%; and for occlusion, 85% and 98%. Above-knee (AK) CTA scores had slightly better concordance of 86.1% than below-knee (BK) readings (82.3%) (p = 0.104). Severe discrepancies between AK CTA and CA scores were observed in 1.8% of segments compared to 5.4% of BK segments (p = 0.038). Poor CTA image quality was the cause in 20% of AK segments and 28% of BK segments. Poor CA image quality was the cause in 8% of AK and 7% of BK discrepancies. Registration disagreement (stenosis observed in a level in 1 study attributed to a different level in the other) accounted for 18% of AK and 17% of BK discrepancies. In 54% of AK and 48% of BK discrepancies, neither image quality nor registration errors were identified, indicating that inherent differences in the depiction of stenosis by CA and CTA were responsible. When discrepancies caused by registration error were excluded, SD observed in BK segments (4.0%) remained significantly higher than in AK segments (1.25%) (p = 0.029), and poor CTA quality image was the most common cause (76%) of severe BK discrepancies. In AK discrepancies without an identifiable technical cause, CTA uniformly showed more stenosis, suggesting greater CTA diagnostic precision in larger vessels. In general, agreement between CTA and CA was moderately good. Compared to CA, CTA may be better at depicting stenosis in large, proximal vessels owing to the superior accuracy of cross sectional images in the measurement of stenosis. There appeared to be poorer CT resolution and higher frequency of severe discrepancies between CTA and CA in BK arteries. PMID- 16382270 TI - Percutaneous isolated limb perfusion with thrombolytics for severe limb ischemia. AB - Patients with severe tibioperoneal disease are poor candidates for a distal bypass. Absence of a distal target, lack of conduit, or multiple medical problems can make these patients a prohibitive risk for revascularization. Acute on chronic ischemia in this group poses a greater challenge. Thrombolytic therapy for acute ischemia can be prolonged and carries a significant risk of bleeding if continued beyond 24 hours. However, if the ischemic limbs can be isolated from the systemic circulation, a higher dose of the lytic agent can be given with lower risk. These are the initial results of a series of 10 patients who underwent percutaneous isolated limb perfusion with a high dose of thrombolytics for severe ischemia. Ten patients (lower extremity 8 and upper extremity 2) presented with severe limb-threatening ischemia. Mean ankle/brachial index (ABI) was 0.15 for the lower extremity, and there were no recordable digital pressures in patients with upper extremity ischemia. No distal target was visible on the initial arteriogram. These patients were then taken to the operating room, and under anesthesia, catheters were placed in an antegrade fashion via femoral approach in the popliteal artery and vein percutaneously. For upper extremity, the catheters were placed in the brachial artery and vein. A proximal tourniquet was then applied. This isolated the limb from the systemic circulation. Heparinized saline was infused through the arterial catheter while the venous catheter was left open. A closed loop or an isolated limb perfusion was confirmed when effluent became clear coming out of the venous port. A high dose of thrombolytic agent (urokinase 500,000 to 1,000,000 U) was infused into the isolated limb via the arterial catheter and drained out of the venous catheter. After 45 minutes, arterial flow was reestablished. In 4 patients, Reopro((R)) was used in addition to thrombolytics. Postprocedure angiograms showed minimal changes, but patients exhibited marked clinical improvement. The ABI changed from 0.15 to 0.5 in the lower extremity and near-normal digital pressures in upper extremity ischemia. Limb salvage and symptomatic relief at 6 months was 90%. All patients except one were kept on anticoagulation postoperatively. No bleeding complications were observed from the procedure. Percutaneous isolated limb perfusion brought symptomatic relief to patients presenting with acute on chronic limb ischemia. This can be an alternate option for patients facing amputation with no revascularization options. PMID- 16382271 TI - Protected carotid artery stenting and angioplasty via transfemoral versus transcervical approaches. AB - This prospective cohort study was taken to determine whether transcervical carotid artery stenting (CAS) with internal carotid artery (ICA) flow reversal is associated with a lower incidence of embolization and femoral access complication when compared with protected, transfemoral CAS in selected, high-risk patients. From 2002 to October 2004, the authors performed 55 carotid stentings and angioplasties. Among the 24 cases via transfemoral approach, 1 developed transient ischemic attack (TIA), 1 stroke, 1 asystolic cardiac arrest, 2 groin hematoma, 2 technical failure, and 1 restenosis. Among the 31 cases via transcervical approach, 2 patients developed TIAs, 4 bradycardia, 2 cervical hematoma, and 3 technical failures leading to open conversion and carotid endarterectomy. Transcervical CAS with ICA flow reversal eliminates the risk of aortic arch emboli, provides cerebral protection during predeployment manipulation across the carotid lesion, negates preprocedure mapping of the aortic arch configuration, and surpasses difficult aortic arch or transfemoral access. PMID- 16382272 TI - Iliofemoral venous pressure correlates with intraabdominal pressure in morbidly obese patients. AB - Clinically, it has been observed that severely and morbidly obese individuals more often have venous leg symptoms related to venous stasis than normal-weight persons have. Obesity is associated with an increased intraabdominal pressure (IAP). The increased IAP in severely and morbidly obese patients would reasonably cause an elevated iliofemoral venous pressure (IFVP), which transmits via incompetent femoral veins, causing venous stasis in the lower limbs. The aim of this study was to determine whether the elevated IAP assessed by the urinary bladder pressure (UBP) corresponded with an increased directly measured IFVP. Fifteen women with morbid obesity were investigated with simultaneous UBP and direct iliofemoral vein pressures. Four normal-weight controls were investigated in the same manner. The obese patients had significantly higher UBP than the controls had, 19.1 and 8.5 cm H2O, respectively. They also had elevated IFVP compared with the controls, 19.7 and 7.5 cm H2O, respectively, and these IFVPs correlated well with the UBPs. The assumption that increased IAP in morbidly obese patients causes increased IFVP was consequently determined. To our knowledge, this has not previously been demonstrated in human individuals. How these elevated pressures contribute to the development of lower limb venous insufficiency is subject to further studies. PMID- 16382273 TI - Risk factors for poor collateral development in claudication. AB - The objective of this retrospective, cross-sectional study was to determine risk factors for poor collateral development in patients with claudication. The authors listed all patients with calf claudication who had undergone angiography in this hospital between 1999 and 2001 and extracted those with superficial femoral artery (SFA) occlusion, a popliteal artery without major lesions, and at least 1 patent calf artery. Forty-five patients met the criteria, and concomitant disease and claudication characteristics, ankle/brachial index (ABI) and number of outflow vessels were recorded. Three blinded observers calculated the number of collaterals on the angiograms, and the collateral count was related to the other factors by use of regression analysis. The mean patient age was 69 years (SD 11), and 62% were women. Their walking distance was 90 m (77) and ABI 0.47 (0.15). Thirty-three percent had diabetes and 50% had duration of symptoms longer than 5 years. The mean number of collaterals bypassing the occlusion was 15.1 (SD 4.8). Univariate regression analysis indicated an association (p <0.08) between few collateral vessels and diabetes, short duration of symptoms, current smoking habits, and old age. In the multivariate analysis only diabetes and short duration of symptoms were related to having few collaterals. In patients with claudication and SFA occlusion, few collaterals from the deep femoral artery appear to be associated with having diabetes and a short duration of symptoms. PMID- 16382274 TI - Embolization of renal artery aneurysm and arteriovenous fistula-a case report. AB - A renal artery aneurysm with an associated arteriovenous fistula in a native kidney has been reported infrequently in the literature. Management depends on size, location, and the patient's physiological condition. We describe a case in which endovascular therapy was used to successfully exclude both aneurysm and fistula. This report describes a 13-centimeter renal artery aneurysm with arteriovenous fistula originating from an isolated branch of the renal artery. Coil-embolization resulted in thrombosis of the aneurysm and fistula while preserving parenchymal perfusion. Coil embolization is an alternative to surgery for coexistent renal artery aneurysm and arteriovenous fistula arising from a branch of adequate length for placement of embolic coils. Successful treatment is not limited by aneurysm size or presence of arteriovenous connection. PMID- 16382275 TI - Salvage of critical limb ischemia with the "trellis reserve'' of subintimal superficial femoral-popliteal artery occlusion: a new modality in managing critical limb ischemia--a case report. AB - Subintimal angioplasty is a safe, effective, but nondurable procedure in treating long superficial femoral artery occlusions in patients with severe lower limb ischemia. The authors report a case of acute thrombosis that presented 16 weeks after subintimal angioplasty. The ;;Trellis'' percutaneous thrombolytic infusion system permitted a controlled site-specific infusion of recombinant tissue-type plasminogen activator (rtPA). The unique design of the ;;Trellis'' allowed complete aspiration of thrombus and avoiding regional and systemic thrombolytic side effects. The ;;Trellis'' system is effective in percutaneous management of thrombotic lesions; however, intimal dissection may need to be addressed. PMID- 16382276 TI - A typology of organizational and contractual arrangements for purchasing and delivery of behavioral health care. AB - The evolution of behavioral health care financing and delivery has led to a wide variety of arrangements connecting consumers to behavioral health services. In this paper, we present a typology based on three distinguishing features of behavioral health arrangements along which there is a high degree of variability and this variability has been shown to affect the cost and quality of behavioral health care: (1) the extent to which sponsor oversight over care is outsourced by way of contracts rather than performed directly; (2) whether financing for behavioral health is partitioned from health care financing overall; and (3) the amount of financial risk shared by the sponsor with third parties. PMID- 16382277 TI - A study on the effect of wet granulation on microcrystalline cellulose particle structure and performance. AB - PURPOSE: The aim of this study was to investigate the mechanism of the effect of wet granulation process on the compaction properties of microcrystalline cellulose (MCC). METHODS: MCC alone and with hydroxypropyl cellulose (HPC) as a binder were wet granulated by a high-shear process using different granulation parameters (over- and undergranulated). Overgranulated batches were also ball milled after drying and compared to the unmilled material. MCC starting material and granulation were characterized for particle size distribution, surface area, porosity, and isothermal moisture uptake. Compaction behavior of the MCC and granulations was also studied using a compaction simulator. RESULTS: In all cases, the wet granulation process decreased MCC primary particle porosity. Wet granulation also reduced compactibility of MCC to different degrees. Overgranulated batch with HPC showed the lowest compactibility and was less compactible than the batch without HPC granulated using the same parameters. Ball milled material showed an increase in porosity and was significantly more compactible than the unmilled granulation from the same batch. CONCLUSIONS: The decrease in MCC compactibility after granulation is associated with the decrease in MCC primary particle porosity and in some cases with the formation of large dense granules as well. Under certain conditions, milling seems to counteract the effect of wet granulation on MCC compactibility. PMID- 16382278 TI - Salt effects on an ion-molecule reaction--hydroxide-catalyzed hydrolysis of benzocaine. AB - PURPOSE: This work investigates the effect of various salts on the rate of a reaction involving a neutral species (benzocaine alkaline hydrolysis). METHODS: Benzocaine hydrolysis kinetics in NaOH solutions in the presence of different salts were studied at 25 degrees C. Benzocaine solubility in salt solutions was also determined. Solubility data were used to estimate salt effects on benzocaine activity coefficients, and pH was used to estimate salt effects on hydroxide activity coefficients. RESULTS: Salts either increased or decreased benzocaine solubility. For example, solubility increased with 1.0 M tetraethylammonium chloride (TEAC) approximately 3-fold, whereas solubility decreased approximately 35% with 0.33 M Na2SO4. Salt effects on hydrolysis rates were more complex and depended on the relative magnitudes of the salt effects on the activity coefficients of benzocaine, hydroxide ion, and the transition state. As a result, some salts increased the hydrolysis rate constant, whereas others decreased it. For example, the pseudo-first-order rate constant decreased approximately 45% (to 0.0584 h(-1)) with 1 M TEAC, whereas it increased approximately 8% (to 0.116 h( 1)) with 0.33 M Na2SO4. CONCLUSIONS: Different salt effects on degradation kinetics can be demonstrated for a neutral compound reacting with an ion. These salt effects depend on varying effects on activity coefficients of reacting and intermediate species. PMID- 16382279 TI - Expression and transport functionality of FcRn within rat alveolar epithelium: a study in primary cell culture and in the isolated perfused lung. AB - PURPOSE: The neonatal constant region fragment receptor (FcRn) binds and transports IgG. FcRn expression in the upper tracheobronchial airways of the lung is recognized. In this study, we sought to characterize the functional expression of FcRn within alveolar regions of lung tissue. METHODS: FcRn immunohistochemistry was performed on intact rat lung. FcRn expression [Western blot, reverse transcription-polymerase chain reaction (RT-PCR), and immunofluorescence microscopy] and IgG transport functionality were assessed in an in vitro rat alveolar epithelial primary cell culture model. An isolated perfused rat lung model was used to examine IgG transport across pulmonary epithelium from airspace to perfusate. RESULTS: FcRn is expressed in intact alveolar epithelium, substantiated by expression and functionality in an in vitro alveolar epithelial model within which IgG transport was temperature sensitive, concentration dependent, and inhibited by excess unlabeled IgG and, to a disproportionate level, by anti-FcRn antibody. Saturable IgG transport across pulmonary epithelium was evident in an isolated perfused rat lung, inhibitable by competing IgG, and displayed a relatively low maximal net IgG absorptive rate of approximately 80 ng/h. CONCLUSION: Pulmonary epithelium expresses functional FcRn providing an absorption pathway potentially important for highly potent Fcgamma fusion proteins but unlikely to be of quantitative significance for the systemic delivery of inhaled therapeutic monoclonal IgGs. PMID- 16382280 TI - Novel mucosal insulin delivery systems based on fusogenic liposomes. AB - PURPOSE: Fusogenic liposomes (FLs) are unique delivery vehicles capable of introducing their contents directly into the cytoplasm with the aid of envelope glycoproteins of Sendai virus (SeV). The objective of this study was to evaluate the potential of FL to improve the mucosal absorption of insulin from rat intestinal membranes. METHOD: The FLs containing insulin were prepared by fusing insulin-loaded liposomes with inactivated SeV particles and were administered directly into the ileal, the colonic, and the rectal loops (10 IU/kg). RESULTS: The FL successfully enhanced the insulin absorption and induced a significant hypoglycemic effect following the colonic and the rectal administration without detectable mucosal damage. This enhancing effect of insulin absorption was further improved by increasing the amount of insulin loaded in the FL and by coencapsulating insulin-degrading enzyme inhibitor. In contrast, the insulin absorption was not increased by the ileal administration of FL because the mucous/glycocalyx layers overlaid on the ileal epithelium impede the fusion of FL to the intestinal mucosa. CONCLUSION: Our results indicated that FL is a useful carrier for improving the absorption of poorly absorbable drugs, such as insulin, via the intestinal tract. PMID- 16382281 TI - Varying the unsaturation in N4,N9-dioctadecanoyl spermines: nonviral lipopolyamine vectors for more efficient plasmid DNA formulation. AB - PURPOSE: The aim of the study is to analyze the effect of varying the degree of unsaturation in synthesized N4,N9-dioctadecanoyl spermines on DNA condensation and then to compare their transfection efficiency in cell culture. METHODS: The N4,N9-di-C18 lipopolyamines-saturated (stearoyl), C9-cis- (oleoyl), and C9,12-di cis- (linoleoyl)-were synthesized from the naturally occurring polyamine spermine. The ability of these novel compounds to condense DNA and form nanoparticles was studied using ethidium bromide fluorescence quenching and nanoparticle characterization techniques. Transfection efficiency was studied in several primary skin cells (FEK4, FCP4, FCP5, FCP7, and FCP8) and in an immortalized cancer cell line (HtTA) and was compared with the commercially available nonliposomal transfection formulation Transfectam (dioctadecylamidoglycyl spermine), which also contains two saturated C18 lipid chains. RESULTS: N4,N9-Dilinoleoyl spermine (C18, di-cis-9,12) is efficient at circular plasmid DNA (pEGFP) condensation and gives the most effective transfection in a series of primary skin cells and cancer cell lines at low charge ratios of 5.5 (+/-ammonium/phosphate). CONCLUSIONS: The dienoic fatty acyl spermine conjugate N4,N9-dilinoleoyl spermine efficiently condenses DNA and achieves the highest transfection levels among the studied lipopolyamines in cultured cells. PMID- 16382282 TI - Bacteria isolated from the different developmental stages and larval organs of the obligate parasitic fly, Wohlfahrtia magnifica (Diptera: Sarcophagidae). AB - Wohlfahrtia magnifica (Diptera: Sarcophagidae) is the major myiasis-causing fly species in the whole of Eurasia for most important domestic animals. The aim of the present work was to obtain data on the culturable bacteria isolated under aerobic conditions from this fly: bacteria were isolated from all developmental stages (larvae, pupa, and imago) of Wohlfahrtia magnifica, and the third-stage larval organs were also sampled. To determine the possible antagonistic effects between the dominant bacterial groups, an antibiosis assay was carried out. Plating and isolation of bacteria was performed by classical microbiological methods. Characterization of the isolated strains was carried out via a polyphasic approach; classical phenotypic tests, chemotaxonomical examinations, and 16S rDNA sequence analyses were also applied. In the case of maggot macerate samples, members of the family Enterobacteriaceae were characteristic. Members of a new genus (Schineria) belonging to the gamma subdivision of proteobacteria were also isolated. According to our data, the shifts in the Schineria and Proteus populations within the larvae are strongly influenced by their interactions with each other and among the members of the family Enterobacteriaceae. The pupa and imago samples contained several other Gram-negative bacteria (Stenotrophomonas, Brevundimonas, etc.). Among Gram-positive bacteria, in all maggot macerate samples, members of the genus Bacillus and the Arthrobacter-Micrococcus group of actinobacteria were dominant (neither of them was a producer or sensitive to the compounds of other microorganisms), and bacteria related to the genus Corynebacterium were also found. From the larvae Aureobacterium liquefaciens and Enterococcus faecalis were isolated, and from the pupae Dietzia maris and Enterococcus faecalis. In the samples of third-stage larval organs, the dominant groups were the same as in the third-stage larval macerate sample; however, several additional genera/species were observed (Rhodococcus fascians, Streptomyces sp., Rathayibacter sp., Bacillus thuringiensis/cereus). PMID- 16382283 TI - Assessment of nitrification potential in ground water using short term, single well injection experiments. AB - Nitrification was measured within a sand and gravel aquifer on Cape Cod, MA, using a series of single-well injection tests. The aquifer contained a wastewater derived contaminant plume, the core of which was anoxic and contained ammonium. The study was conducted near the downgradient end of the ammonium zone, which was characterized by inversely trending vertical gradients of oxygen (270 to 0 microM) and ammonium (19 to 625 microM) and appeared to be a potentially active zone for nitrification. The tests were conducted by injecting a tracer solution (ambient ground water + added constituents) into selected locations within the gradients using multilevel samplers. After injection, the tracers moved by natural ground water flow and were sampled with time from the injection port. Rates of nitrification were determined from changes in nitrate and nitrite concentration relative to bromide. Initial tests were conducted with (15)N enriched ammonium; subsequent tests examined the effect of adding ammonium, nitrite, or oxygen above background concentrations and of adding difluoromethane, a nitrification inhibitor. In situ net nitrate production exceeded net nitrite production by 3- to 6- fold and production rates of both decreased in the presence of difluoromethane. Nitrification rates were 0.02-0.28 mumol (L aquifer)(-1) h(-1) with in situ oxygen concentrations and up to 0.81 mumol (L aquifer)(-1) h(-1) with non-limiting substrate concentrations. Geochemical considerations indicate that the rates derived from single-well injection tests yielded overestimates of in situ rates, possibly because the injections promoted small-scale mixing within a transport-limited reaction zone. Nonetheless, these tests were useful for characterizing ground water nitrification in situ and for comparing potential rates of activity when the tracer cloud included non-limiting ammonium and oxygen concentrations. PMID- 16382284 TI - The effects of cyanobacterial exudates on bacterial growth and biodegradation of organic contaminants. AB - The pulp and paper industry largely depends on the biodegradation activities of heterotrophic bacteria to remove organic contaminants in wastewater prior to discharge. Our recent discovery of extensive cyanobacterial communities in pulp and paper waste treatment systems led us to investigate the potential impacts of cyanobacterial exudates on growth and biodegradation efficiency of three bacterial heterotrophs. Each of the three assessed bacteria represented different taxa commonly found in pulp and paper waste treatment systems: a fluorescent Pseudomonad, an Ancylobacter aquaticus strain, and a Ralstonia eutropha strain. They were capable of utilizing phenol, dichloroacetate (DCA), or 2,4 dichlorophenoxyacetic acid (2,4-D), respectively. Exudates from all 12 cyanobacterial strains studied supported the growth of each bacterial strain to varying degrees. Maximum biomass of two bacterial strains positively correlated with the total organic carbon content of exudate treatments. The combined availability of exudate and a known growth substrate (i.e., phenol, DCA, or 2,4 D) generally had a synergistic affect on the growth of the Ancylobacter strain, whereas mixed effects were seen on the other two strains. Exudates from four representative cyanobacterial strains were assessed for their impacts on phenol and DCA biodegradation by the Pseudomonas and Ancylobacter strains, respectively. Exudates from three of the four cyanobacterial taxa repressed phenol biodegradation, but enhanced DCA biodegradation. These dissimilar impacts of cyanobacterial exudates on bacterial degradation of contaminants suggest a species-specific association, as well as a significant role for cyanobacteria during the biological treatment of wastewaters. PMID- 16382285 TI - A phase of increased ST elevation during coronary occlusion following ischemic preconditioning. AB - ATP-sensitive potassium channels are opened during the course of ischemic preconditioning (IP). As experimental data suggest that opening of sarcolemmal ATP-sensitive potassium channels underlie ST elevation during myocardial ischemia, one would expect to observe increased ST elevation during ischemia following IP. However, clinical studies have reported IP to attenuate ST elevation during repeated brief coronary occlusions. The objective of this study was to characterize the temporal course of ST elevation during coronary occlusion following IP. Twenty-eight closed-chest pigs were subject to catheter-based left anterior descending coronary artery occlusion/ reperfusion for 45/120 minutes. Thirteen animals were preconditioned by two occlusion/reperfusion cycles of 10/30 minutes. Fifteen pigs served as controls. The electrocardiographic ST vector magnitude was continuously monitored. IP reduced the infarct size normalized for area at risk (IP 9.6 +/- 15.8%; control 71.2 +/- 14.7%; p < 0.001). IP increased the time between coronary artery occlusion and appearance of significant rise in ST vector magnitude from 51 +/- 17 to 94 +/- 33 seconds (p < 0.01). IP reduced the rise in ST vector magnitude after 120 seconds of occlusion from 202 +/- 85 microV to 68 +/- 28 microV (p < 0.001) and increased the rise in ST vector magnitude after 600 seconds from 265 +/- 106 microV to 427 +/- 232 microV (p < 0.001). CONCLUSION: Ischemic preconditioning reduced and delayed early ST elevation during subsequent coronary artery occlusion, but increased late ST elevation. Thus, ischemic preconditioning causes a dynamic and critically time dependent biphasic pattern of ST elevation during repeated coronary occlusions. PMID- 16382286 TI - Influence of short-term versus prolonged cardiopulmonary receptor stimulation on renal and preganglionic adrenal sympathetic nerve activity in rats. AB - Renal and preganglionic adrenal sympathetic nerve activities (RSNA, ASNA) are regulated differentially. Various cardiopulmonary receptor (CPR) stimulation procedures were performed to distinguish short-term and prolonged as well as mechanical and chemical stimulatory effects on RSNA and ASNA. In anesthetized male Sprague-Dawley rats blood pressure, heart rate, left ventricular end diastolic pressure (LVEDP), RSNA and ASNA were recorded. CPRs were stimulated as follows: Short-term mechanical: LVEDP changes (+/-4, +/-6, +/-8 mmHg) via aortic and caval vein occlusion; Short-term chemical: phenylbiguanide (PBG-bolus, 0.1, 1, 10 microg IV); Prolonged mechanical (15 min): volume expansion (0.9% NaCl, 5% body weight) and hemorrhage, to modulate LVEDP; Prolonged chemical: PBG infusion (32 microg/min IV, for 15 min); Stimulations were done with 1) all afferents intact, 2) bilateral cervical vagotomy (VX), 3) VX + SAD (sino-aortic denervation; short-term protocols and hemorrhage).1) Short-term mechanical stimuli decreased RSNA (-52 +/- 12%) and ASNA (-37 +/- 13%). 2) PBG-bolus decreased RSNA (-54 +/- 12%) but increased ASNA (+40 +/- 13%). 3) Volume expansion decreased RSNA (-55 +/- 7%), ASNA was unaffected. 4) PBG infusion persistently decreased RSNA (-60 +/- 6%) but just shortly increased ASNA (+120 +/ 15%); VX abolished all responses. 5) Hypotensive hemorrhage decreased RSNA (-39 +/- 9%) but increased ASNA (+42 +/- 9%). VX abolished RSNA response; ASNA response only disappeared with VX + SAD.Short-term mechanical CPR stimulation uniformly decreased sympathetic activities, whereas chemical stimulation had opposing effects on renal and adrenal sympathetic responses. All prolonged stimuli decreased RSNA, whereas ASNA was virtually unaffected: Sympathetic out.ow is differentially controlled not only with regard to target organs or afferent receptors but also stimulus time pattern. PMID- 16382287 TI - ACE-inhibitor treatment attenuates atrial structural remodeling in patients with lone chronic atrial fibrillation. AB - OBJECTIVE: Chronic atrial fibrillation (AF) is characterized by a remodeling process which involves the development of fibrosis. Since angiotensin II has been suspected to be involved in this process, the aim of our study was to investigate a possible influence of an ACE-I therapy in patients with chronic AF regarding the occurrence of left atrial structural remodeling. METHODS: Atrial tissue samples were obtained from patients with lone chronic AF or sinus rhythm (SR). Collagen I, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) protein expression were measured by quantitative Western Blotting techniques and calculated as mean +/- SEM. Histological tissue samples were used for calculating microvessel density (microvessel/mm(2) +/- SEM). RESULTS: In AF, the collagen amount was higher (1.78 +/- 0.21; p = 0.01) vs. SR (0.37 +/- 0.07) accompanied by declining microcapillary density (AF: 145 +/- 13 vs. SR: 202 +/- 9; p = 0.01). Additionally, a negative correlation (p = 0.01) between collagen content and microcapillary density was observed. To investigate the influence of an ACE-I therapy on this remodeling process, patient groups were divided into AF and SR both with or without ACE-I. Interestingly, there was a significantly lower expression of collagen I in AF with ACE-I (1.04 +/- 0.26) vs. AF without ACE-I treatment (2.07 +/- 0.24, p = 0.02). The microcapillaries were not diminished in AF with ACE-I (180 +/- 15) vs. SR with ACE-I (196 +/- 9), but there was a significant rarification in AF without ACE-I (123 +/- 18; p = 0.03). The expression of VEGF and bFGF did not reveal any significant differences. CONCLUSION: In patients undergoing ACE-I treatment: atrial structural remodeling was attenuated and the loss of atrial microcapillaries was prevented. PMID- 16382288 TI - The therapeutic potential of progenitor cells in ischemic heart disease--Past, present and future. AB - Within the recent past, it has become increasingly evident that adult bone marrow and peripheral blood do not only contain diverse hematopoietic precursors but also stem- and progenitor cells with some properties resembling those of embryonic cells. These adult progenitor cells can contribute to the regeneration of injured tissue as has been revealed by in vitro experiments and subsequent animal studies. Over the last four years, several clinical studies employing such a hypothesis in the context of myocardial repair after acute infarction or during chronic ischemic failure have been published. These studies have used autologous bone marrow cells as well as peripheral blood-derived progenitor cells, which were delivered via intracoronary or intramyocardial routes near the infarct area. The initial results demonstrated the safety and possible benefit of this strategy, which appear to be relatively simple, inexpensive and free of side effects. However, the present clinical studies were small in size so that the overall therapeutic efficacy remains open to debate and evaluation. Furthermore, a major part of the molecular mechanisms underlying progenitor cell-mediated repair has been proposed but not yet elaborated. Hence, larger controlled randomized and double-blinded trials in addition to experimental investigations on the primary mechanisms of repair are keenly anticipated. PMID- 16382289 TI - Effect of catheter-based transendocardial delivery of stromal cell-derived factor 1alpha on left ventricular function and perfusion in a porcine model of myocardial infarction. AB - BACKGROUND: Myocardial regeneration after myocardial infarction can occur via stem cell recruitment. Stromal cell-derived factor 1alpha (SDF-1alpha) has been shown to be critical for stem cell homing to injured tissue. METHODS: Myocardial infarction was induced in pigs via microembolization of the distal left anterior descending artery. Two weeks after myocardial infarction animals underwent catheter-based transendocardial injection of SDF-1alpha into the periinfarct myocardium (18 injections, 5 ig per injection) (n = 12) or sham-intervention (n = 8). Tc99m sestamibi single-photon emission computed tomography (SPECT) and electromechanical mapping (EMM) of the left ventricle were performed two and seven weeks after myocardial infarction. RESULTS: Infarct size by tetrazolium staining was similar in both groups (8.9 +/-1.2% of left ventricle vs. 8.9 +/- 2.6%). Vessel density in the periinfarct area was significantly higher in SDF 1alpha treated animals than in controls (349 +/- 17/mm2 vs. 276 +/- 21/mm2, p < 0.05). Myocardial perfusion (SPECT) did not change in either group. Ejection fraction and stroke volume (EMM) decreased in SDF-1alpha animals and increased in controls (difference between groups p = 0.05 for ejection fraction and p < 0.05 for stroke volume). Linear local shortening (EMM) did not change in controls (11.4 +/- 1.3% to 11.5 +/- 0.5%) but decreased significantly in SDF-1alpha treated animals (12.1 +/- 0.9% to 8.4 +/- 0.9%, p < 0.05, p < 0.05 for difference between groups). SDF-1 delivery was associated with a substantial loss of collagen in the periinfarct area (32+/-5% vs. 61+/-6% in control animals, p < 0.005). CONCLUSION: A strategy to augment stem cell homing by catheter-based transendocardial delivery of SDF-1alpha in experimental myocardial infarction increases periinfarct vessel density, fails to improve myocardial perfusion, is associated with loss of collagen in the periinfarct area and impairs left ventricular function. PMID- 16382290 TI - Commentary on "Direct comparison of magnetic resonance imaging and conductance microcatheter in the evaluation of left ventricular function in mice" by Jacoby et al. in Basic Res Cardiol 101:87-95 (2006). PMID- 16382292 TI - Allopurinol or oxypurinol in heart failure therapy - a promising new development or end of story? AB - The plasma level of the uric acid is frequently elevated in heart failure, due to increased production and/or to reduced renal excretion of this antioxidant metabolite. The transformation of hypoxanthine to xanthine and the conversion of the latter into uric acid, which occur in purine catabolism, are catalysed by xanthine oxidoreductase. The constitutive xanthine dehydrogenase form of this enzyme generally uses NAD(+) as an electron acceptor, whereas the post translational xanthine oxidase form uses molecular oxygen and yields four units of reactive oxygen species per unit of transformed substrate. Allopurinol and oxypurinol inhibit xanthine oxidoreductase and thus diminish the generation of reactive species and decrease plasma uric acid. In a recent study in patients with NHYA class II-III heart failure, add-on treatment with allopurinol 300 mg/day for 3 months lowered plasma uric acid but failed to improve laboratory exercise performance or the distance walked in 6 minutes. In another recent trial, which was carried out in patients with NHYA class III-IV heart failure, add-on treatment with oxypurinol 600 mg/day for 24 weeks decreased plasma uric acid concentration but did not change a composite of patient outcome and state. These results indicate that the reduction in plasma uric acid caused by allopurinol or oxypurinol does not benefit patients with heart failure. Moreover, the hypothesis that the diminution in the renal excretion of the antioxidant uric acid caused by diuretics may be salutary in cardiac failure is strengthened by the study results considered. PMID- 16382293 TI - Cardioprotective-mimetics reduce myocardial infarct size in animals resistant to ischemic preconditioning. AB - BACKGROUND: Ischemic preconditioning (IPC) elicits two distinct windows of cardioprotection, an early phase that lasts for 1-2 h and a delayed phase that lasts for 24-72 h. However, there is conflicting data as to how long the heart is resistant to IPC-induced cardioprotection after the initial protection wanes, leading to the demonstration of IPC-resistance. This resistance to IPC appears to be dependent on the timing of the next IPC stimulus, the species of animals used and the model studied. Furthermore, the mechanisms responsible IPC-resistance are unknown. It is also important to demonstrate therapeutic interventions that will produce cardioprotection during this period of IPC-resistance. METHODS AND RESULTS: To examine potential mechanisms responsible for acute IPC-induced resistance, the NHE-1 inhibitor EMD 85131 (2-methyl-5-methylsulfonyl-1-(1 pyrrollyl)-benzoylguanidine), which exerts its effects via mechanisms distinct from IPC, and the K(ATP) channel opener bimakalim, which bypasses the signaling mechanisms of IPC to directly open K(ATP) channels, were examined in a canine model of IPC-resistance. One 10 min. IPC stimulus followed by 10 min. of reperfusion produced a significant reduction in IS/AAR compared to Control (7.1 +/- 2.6% versus 26.0 +/- 6.2%; P < 0.05). However, IPC did not significantly protect the myocardium if a 2 h reperfusion period occurred between the initial IPC stimulus and the subsequent prolonged (60 min) ischemic challenge (IS/AAR: 22.5 +/- 4.8%: P > 0.05). Furthermore, hearts treated with IPC followed by 2 h of reperfusion were resistant to an additional IPC stimulus administered just prior to the subsequent 60 min. occlusion period (IS/AAR: 22.9 +/- 3.2%: P > 0.05). In contrast, administration of the NHE-1 inhibitor EMD 85131 (IS/AAR: 7.4 +/- 2.5%: P < 0.05) or the K(ATP) channel opener bimakalim (IS/AAR: 11.8 +/- 2.4%: P < 0.05) both afforded significant cardioprotection when administered at 2 h of reperfusion in previously preconditioned canine hearts resistant to IPC. CONCLUSIONS: IPC resistance occurs in this canine model of ischemia-reperfusion injury. However, in spite of IPC resistance, hearts can still be pharmacologically protected by direct application of the K(ATP) channel opener bimakalim or the NHE inhibitor EMD 85131. PMID- 16382294 TI - Vasopeptidase inhibition peri- and post-MI in Zucker insulin resistant rats: effect on MI size, arrhythmias, remodeling, function and fetal gene expression. AB - Mortality peri-myocardial infarction (MI) is increased with insulin resistance. As the vasopeptidase inhibitor (VPI) omapatrilat improves insulin sensitivity, it may be beneficial peri-MI in Zucker Insulin Resistant rats (ZIR). ZIR rats (n = 228) received omapatrilat 10 mg/kg/day, 7 days pre-MI, to 38 days post-MI, or control. Twenty-four protocol (n = 72): a subgroup of rats received the kinin receptor antagonist icatibant. Ambulatory ECG recordings, and MI size were evaluated. Thirty-eight-day protocol (n = 156): left ventricular (LV) remodeling, cardiac hemodynamics, morphology, infarct size, and RT-PCR for GLUT-4 and fetal genes were measured. Omapatrilat improved post-MI survival 24 h (62% vs 38%, P = 0.0007) which was maintained 38 days. There was a kinin-induced reduction of ventricular arrhythmias and there appeared to be a kinin-independent reduction in MI size (23.5 +/- 2.4% vs 17.0 +/- 2.2%, P = 0.053) for 24-h post-MI. Omapatrilat reduced but did not prevent LV dilatation, dysfunction, and fetal gene expression 38 days post-MI. Omapatrilat did not prevent reduced cardiac GLUT-4 expression. In ZIR rats, mortality post-MI is reduced by omapatrilat, due and a kinin dependent reduction in ventricular arrhythmias and possibly a kinin-independent reduction in MI size. Ventricular dilatation, dysfunction, and fetal gene expression are variably attenuated but not prevented. PMID- 16382295 TI - Chronic erythropoietin treatment limits infarct-size in the myocardium in vitro. AB - It has been well established that erythropoietin (EPO) can limit myocardial ischemia/reperfusion injury in a variety of acute settings. However, despite EPO being used chronically to treat anemia the infarct limiting effects of long term treatment (chronic) have never been fully investigated. In this study we examined the effects of a 3 week treatment of EPO (5,000 IU/Kg) in male Sprague Dawley rats in limiting myocardial infarction after 35 min ischemia and 2 h reperfusion in an in vitro isolated heart perfusion model. Treating the animals 'once a week' failed to limit infarct size significantly compared to a saline control (54.1% +/ 3.5 v 52.3% +/- 4.4), whereas a '3 times a week' regime succeeded in significantly reducing infarct size (36.2% +/- 3.2 v 52.3% +/- 4.4, p < 0.05). To demonstrate that the effect was not due to improved oxygen supply caused by a raised hematocrit level, we also administered EPO 24 h prior to ischemia/reperfusion. This treatment again reduced infarct size compared to a saline control (39.9% +/- 4.4 v 58.4% +/- 5.0, p < 0.05). To examine the mechanism of protection we used the PI3K inhibitor wortmannin and the nitric oxide synthase inhibitor L-NAME to try to abrogate EPO mediated protection. Where wortmannin failed to block the effects of EPO (31.7% +/- 6.0 v 36.2% +/- 3.2), L NAME did abrogate protection (51.6% +/- 5.6 v 36.2% +/- 3.2, p < 0.05). We demonstrate that chronic EPO treatment limits infarct size and that it does so in a nitric oxide dependent manner. PMID- 16382296 TI - The sulfonylurea glipizide does not inhibit ischemic preconditioning in anesthetized rabbits. AB - The K(ATP) channel blocker glibenclamide inhibits cardioprotection afforded by ischemic preconditioning (IPC), raising concern about sulfonylurea use by patients with cardiovascular disease. We examined the effects of the widely prescribed sulfonylurea glipizide (Glucotrol XL(R) ) on IPC in anesthetized rabbits. Initially, in parallel studies in pentobarbital-anesthetized rabbits, we identified doses of glipizide (GLIP, 0.17 mg/kg + 0.12 mg/kg/h, IV) and glibenclamide (GLIB, 0.05 mg/kg + 0.03 mg/kg/h, IV) that produced steady-state, clinically relevant plasma levels of both drugs; these doses also significantly increased plasma insulin by 51 +/- 17% (GLIP) and by 57 +/- 17% (GLIB, both p < 0.05 vs. their respective baseline levels). Subsequent parallel studies in ketamine-xylazine-anesthetized rabbits examined the effects of these doses of GLIP and GLIB on IPC. Myocardial injury (30 min coronary occlusion/120 min reperfusion), either with or without IPC (5 min occlusion/10 min reperfusion) was induced midway during a 2 h infusion of vehicle (VEH), GLIP or GLIB (n = 10-11 each). Infarct area (IA) normalized to area-at-risk (%IA/AAR) was 62 +/- 3% in the VEH group, and was significantly reduced to 39 +/- 5% by IPC (p < 0.05 vs. VEH). Neither GLIP nor GLIB treatment had any effect on %IA/AAR in the absence of IPC (p > 0.05). IPC-induced cardioprotection was preserved in the GLIP + IPC treatment group (45 +/- 4%) when compared to VEH alone (p < 0.05), but was attenuated in the presence of GLIB (GLIB+IPC: 53 +/- 4% IA/AAR, p > 0.05 vs. VEH). Thus, at a clinically relevant plasma concentration, glipizide did not limit the cardioprotective effects of IPC, and is unlikely to increase the severity of cardiac ischemic injury. PMID- 16382297 TI - Carvedilol exerts more potent antiadrenergic effect than metoprolol in heart failure. AB - BACKGROUND: It is still uncertain whether or not there is a difference between metoprolol and carvedilol for the treatment of congestive heart failure. We attempted to determine the difference between the two beta-blockers in terms of their antiadrenergic effect during exercise in patients with heart failure and their efficacy based on the baseline plasma brain natriuretic peptide concentration. METHODS: Fifty-three patients with mild to moderate heart failure with a radionuclide left ventricular ejection fraction <40% received open label metoprolol or carvedilol in a randomized fashion. The increase in the heart rate normalized to the increase in the plasma norepinephrine concentration during exercise, was calculated as an index of adrenergic responsiveness during exercise. RESULTS: The increase in heart rate normalized by the increase in plasma norepinephrine concentration, decreased after the initiation of beta blockers in the carvedilol group, but not in the metoprolol group. The change in cardiac function was more favorable for carvedilol than metoprolol in patients who exhibited a higher baseline brain natriuretic peptide concentration. CONCLUSIONS: Carvedilol exerts a more potent antiadrenergic effect than metoprolol during stress in patients with mild to moderate heart failure. Carvedilol appears to be more efficacious than metoprolol in patients who exhibit higher baseline brain natriuretic peptide concentrations. These differences should be kept in mind when selecting appropriate pharmacologic agents in the treatment of heart failure. PMID- 16382298 TI - Dipyridamole, an underestimated vascular protective drug. AB - Dipyridamole, developed almost half a century ago, acts by inhibiting nucleoside transport, which increases adenosine levels leading to inhibition of platelet aggregation and vasodilatation mainly in the coronary tree. It is a vaso protective drug with proven efficacy in the prevention of strokes. Adenosine receptor 2 inhibitory purines, ubiquitously available in food and drink, inhibit the vasomotor effects of dipyridamole but not its action on platelet aggregation. This and the slow build-up of blood levels of dipyridamole after oral application may explain why incidents of drug-induced angina ("coronary steal") were never reported in the prevention trials. The prevention of arterial thrombosis and the positive remodeling of the arterial system (arteriogenesis) by elevated blood flows suggest that dipyridamole may be able to halt the progression of organ manifestations of atherosclerosis. Clinical trials for the secondary prevention of vascular occlusions in other vascular beds should be encouraged. PMID- 16382299 TI - Economic impact of hyperhomocysteinemia in Switzerland. AB - Since more than 20 years elevated homocysteine plasma levels have been associated with an elevated cardiovascular risk. It can be assumed that approx. 5-7% of the Swiss population suffers from hyperhomocysteinemia. These people have an odds ratio of 1.7 (95% confidence interval: 1.5-1.9) to develop a myocardial infarction and an odds ratio of 2.5 (95% confidence interval 2-3) of developing a stroke. These significant cardiovascular endpoints have monetary implications and lead to a loss in life years. The cost consequences and total life years lost were determined with an incidence-based epidemiological model utilizing a Swiss third party payer perspective. We could demonstrate that hyperhomocysteinemia related sequelae (myocardial infarction and stroke) amount to 41.1-110.2 million CHF. In addition it can be estimated that 6'941-18'478 life years may be lost. Comparing these data with the total costs for cardiovascular disease in Switzerland of CHF 987 million, we estimate the share of the economic burden of hyperhomocysteinemia at approximately 10%. Preventive measures could thus yield a positive impact on total health care expenditure in the Swiss healthcare system and warrants further research. PMID- 16382300 TI - Intracranial hemorrhages associated with intravenous platelet glycoprotein IIB/IIIA receptor inhibitors in the United States. AB - OBJECTIVES: To determine the rates of intracranial hemorrhages associated with GP IIb/IIIa inhibitors in routine practice. BACKGROUND: Rates of intracranial hemorrhages (ICH) among patients treated with platelet glycoprotein (GP) IIb/IIIa inhibitors for coronary interventions and acute coronary syndromes have been studied within clinical trials but not in routine practice. METHODS: We evaluated the rates of ICH in routine practice in United States (US) using national estimates of rates, in-hospital outcomes, and mortality obtained from National Hospital Discharge Survey. RESULTS: There were 367 294 patients aged 18 years or greater who were treated with platelet GP IIb/IIIa inhibitors between 2000 and 2002 in United States. ICH was observed in 479 (0.13%) of the 367,294 patients with a 100% associated mortality. ICHs related to GP IIb/IIIa inhibitors comprised 0.12% of the total number of ICHs (n = 411 621) observed in United States between 2000 and 2002. CONCLUSIONS: ICH related to platelet GPIIb/IIIa inhibitors is uncommon but associated with high mortality. PMID- 16382301 TI - An improved method of 90% pancreatectomy using a low dose of streptozotocin at the pancreaticoduodenal artery results in a rapid diabetic stage in dogs. AB - An improved method of inducing diabetes in dogs was developed. This method included 90% pancreatectomy, 2 mg/kg streptozotocin (STZ) perfused into pancreaticoduodenal artery, and the fixation suture of the duodenum to the costo abdominal wall. Vasopressin injection administered to the animals before surgery reduced bleeding. All dogs used in this procedure survived and became diabetic. One month after the procedure the pancreatic islets were reduced in volume and the number compared with pancreas tissue obtained during the surgery. Acinar tissue remained with a normal histology, and exocrine function maintained the physiological parameters, except for a soft faecal consistency. We conclude that this procedure is effective in inducing experimental diabetes in dogs. PMID- 16382302 TI - Metabolic control and educational status in children with type 1 diabetes: effects of a summer camp and intensive insulin treatment. AB - Our aim was to evaluate prospectively, in our diabetic patients, the impacts of a summer camp and intensive insulin treatment (IIT) on both metabolic control and disease-related educational level. Twenty-five patients participated in a 7-day long summer camp. Before the camp, all patients were on therapy with short-acting human insulin (SAI) and intermediate-acting insulin (IAI) twice daily. On arrival, their insulin therapy regimen was changed by IIT including either SAI or rapid-acting insulin analogue (RAI) three times before meals supplemented by IAI at bedtime. Following the camp, all participants were given IIT with RAI plus IAI. Frequency of hypoglycaemia, insulin dose, body mass index (BMI) and glycohaemoglobin (HbA1c) levels were assessed at pre-camp and post-camp controls. To evaluate the effectiveness of camp-assisted education, all participants were regularly tested. We observed significant elevations in total daily dose of insulin and BMI at months 3 and 6 when compared with the pre-camp values but, by month 12, they were not significantly different from precamp values. The mean HbA(1c) level decreased significantly at months 6 and 12. Severe hypoglycaemic episodes and ketoacidosis were not detected during the camp and the following year. Significant improvements in knowledge about diabetes and self-management were determined at the end of the camp, after 6 and 12 months. Camp-assisted IIT with RAI improved metabolic control of diabetic children. Additionally, camp assisted education has a positive effect on disease-related educational level and self-management. PMID- 16382303 TI - Insulin resistance in the Zucker diabetic fatty rat: a metabolic characterisation of obese and lean phenotypes. AB - The Zucker diabetic fatty (ZDF) rat is a commonly used animal model of type 2 diabetes yet complete descriptions of insulin resistance in this model are limited. We present a full characterisation of in vivo insulin resistance in obese (fa/fa) animals compared to lean (+/?) littermates. Anaesthetised, ten-week old, obese ZDF rats and their lean littermates underwent a hyperinsulinaemic euglycaemic glucose clamp. Compared with lean littermates, obese ZDF rats required an 89% lower glucose infusion rate to maintain euglycaemia and showed a 35% decrease in peripheral glucose disposal. Insulin-stimulated glucose uptake (R(g')) in obese animals was also significantly less in all skeletal muscles studied. R(g') in cardiac and white adipose tissue was not different between the two groups. Total glycogen content in skeletal and cardiac muscle was significantly less in obese animals, while total glycogen content in the liver was significantly greater than in lean littermates. Glycogen synthesis was also decreased in skeletal muscle of obese animals. Compared with lean animals, total triglyceride content was significantly greater in skeletal muscle, heart and liver of obese ZDF rats. Obese animals also showed significantly increased glucose incorporation into lipid in all of these tissues, indicating an increase in lipogenesis. Collectively, these results provide an integrated characterisation of in vivo insulin resistance in obese ZDF rats and a direct comparison with lean littermates. PMID- 16382304 TI - Cardiac function and ischaemic tolerance during acute loss of metabolic control in the diabetic BB Wor rat. AB - The effect of loss of metabolic control, by with-holding insulin treatment, on reperfusion recovery of cardiac function following ischaemia was studied in the spontaneously diabetic "BB" Wor rat. The study involved a group of insulin treated diabetic BB rats (insulin-treated) and diabetic BB rats in which insulin treatment was withheld 24 h prior to study (insulin-withdrawn). Hearts were isolated and perfused at a constant left atrial filling pressure of 15 cm H(2)O and aortic afterload resistances of 100 and 140 cm H(2)O. Hearts were then subjected to 20 min of ischaemia followed by 30 min of reperfusion. Withdrawing insulin treatment from the BB Wor rat resulted in a dramatic increase in the levels of plasma glucose and free fatty acids. Hearts from these rats perfused under aerobic conditions demonstrated reductions in heart rate, positive and negative dP/dt, cardiac output and left ventricular minute work, whereas diastolic pressure was elevated. Following ischaemia, recovery of cardiac function in the insulin-treated BB Wor rat returned to preischaemic levels, whereas hearts from insulin-withdrawn rats displayed impaired recovery. Throughout reperfusion, heart rate, positive dP/dt, cardiac output and left ventricular minute work remained significantly lower in hearts from insulin withdrawn rats compared to treated rats. Our results indicate that acute loss of metabolic control increases the sensitivity of the heart to ischaemia, even in the acutely diabetic BB Wor rat. PMID- 16382305 TI - Daily profile of plasma %CoQ10 level, a biomarker of oxidative stress, in patients with diabetes manifesting postprandial hyperglycaemia. AB - The purpose of this study was to investigate the effect of regular meals on the daily profile of blood oxidative stress markers in type 2 diabetic patients with postprandial hyperglycaemia. %CoQ10, calculated as the ratio of ubiquinone-10 (oxidised form of coenzyme Q10) to ubiquinol-10 (reduced form), was used as a sensitive marker of oxidative stress. Blood samples were collected from patients before and 2 h after breakfast, lunch and supper, and at 10 p.m. Patients were selected for the study if their blood glucose levels were <7 mmol/l before breakfast and > or =11.1 mmol/l on at least one occasion after breakfast. %CoQ10 levels after breakfast and throughout the day were significantly higher than those before breakfast (p=0.006-0.04). In contrast to the wave-like changes in plasma glucose levels, %CoQ10 levels increased after breakfast and remained at high levels throughout the day. These results indicated that diabetic patients with postprandial hyperglycaemia were exposed to meal-induced periods of oxidative stress during the day. Postprandial hyperglycaemia therefore has the potential to increase the risk of atherosclerotic cardiovascular disease through induction of oxidative stress. PMID- 16382306 TI - Use of insulin glargine in patients with hyperglycaemia receiving artificial nutrition. AB - The purpose of this study was to verify whether it is possible to use insulin glargine (Lantus) subcutaneously in patients receiving artificial nutrition (AN) and if the analogue is capable of obtaining and maintaining good glycaemic control without inducing hypoglycaemia. The sample considers 25 patients receiving AN, diagnosed diabetics and non-diabetics, who had previously been prescribed traditional insulin therapy. All were to be given subcutaneous insulin glargine at a dosage equal to the average of insulin/day administered in the preceding days spent receiving AN. Twenty-five consecutive patients, not stratified in any way, were judged eligible in the last six months of 2004 and first two months of 2005. Four out of these 25 could not be evaluated, either because they did not begin or complete the treatment with Lantus (3/4) or because the proper number of glycaemic tests were not carried out (1/4); 21/25 patients, 84% of the sample with a mean age of 68.7 years (range 46-91 years), finished the study and could be evaluated. The mean glycaemic values after treatment with glargine were already better on the second day, and on the seventh day the difference was statistically significant. No hypoglycaemia requiring medical intervention occurred. This study confirms the possibility of using insulin glargine in patients receiving AN with hyperglycaemia regardless of the type of nutrition and whether or not the patient is diabetic. PMID- 16382307 TI - Sources of plasma glucose and liver glycogen in fasted ob/ob mice. AB - Alterations in intrahepatic carbohydrate fluxes in ob/ob mice and the effects of acute leptin administration were studied in vivo by use of a dual-isotope tracer infusion. Metabolic sources of plasma glucose (gluconeogenesis (GNG) and glycogenolysis) and hepatic glycogen (GNG, direct synthesis and pre-existing) were determined in 20-h-fasted mice infused with [2-13C1]glycerol and [U13C6]glucose for 3 h. Total glucose output (TGO) and the rate of appearance (Ra) of plasma glycerol were measured by isotope dilution. GNG, the direct pathway of hepatic glycogen synthesis and hepatic triose-phosphate flux were determined by mass isotopomer distribution analysis (MIDA). Serum glucose, insulin, leptin and liver glycogen concentrations were also measured. After a 24 h fast, ob/ob mice had 2-fold higher TGO, 2.5-fold elevated liver glycogen content and markedly higher glycogenolytic flux to glucose, absolute GNG and direct glycogen synthesis rates (10-fold increased) compared to the control group. Ob/ob mice also had elevated triose-phosphate flux compared to controls (40 vs. 22 mg/kg lean body mass/min). A model of intrahepatic flux distributions in control and ob/ob mice is presented. In summary, elevated fasting plasma glucose concentrations are due to increased TGO in ob/ob mice, which is maintained by both increased GNG and increased glycogenolysis. Furthermore, the ob/ob mice have major alterations in fasting hepatic carbohydrate fluxes into triose-phosphate pools and glycogen. We support the model that actions of leptin on hepatic glucose metabolism require insulin or other factors. PMID- 16382308 TI - Pannus resolution after occipitocervical fusion in a non-rheumatoid atlanto-axial instability. AB - Periodontoid pseudotumor or pannus is considered to be an inflammatory mass most frequently associated with rheumatoid arthritis. Transoral resection of the pannus has been the treatment of choice for patients with associated myelopathy, followed in many instances by posterior stabilization. However, some authors have reported resolution of pannus associated with rheumatoid arthritis and other forms of chronic atlanto-axial instability only after posterior stabilization. We report a case of a 69-year-old man who presented with a rapidly progressing myelopathy due to a retro-odontoid mass produced by chronic atlanto-axial instability associated with an occipital assimilation of C1 and tight posterior fossa. An urgent posterior fossa craniectomy followed by occipitocervical fixation was performed. After surgery, the patient's clinical condition improved and 1 year after surgery was asymptomatic, walked without any help and had normal strength. Control MR showed complete resolution of the retro-odontoid pannus. PMID- 16382309 TI - The influence of vertebral instability on peridural circulation and concomitant peridural fibrosis formation. AB - An animal model of vertebral instability was used to analyze the effect of chronic lumbar instability on the peridural vasculature and fibrosis formation. Fifty mature male domestic rabbits were divided into five equal groups. The vertebral instability was performed by excision of supra and interspinous ligaments between L2-L3 and L3-L4, excision of transverse and spinous processes and making bilateral laminectomies and facetectomies in groups I, II, III and IV. In group V only para vertebral muscle dissection was performed without vertebral instability. The simulation of the long term effects of overuse model on unstable spines (chronic instability) were performed with the use of Electrical Neuromuscular Stimulator to simulate cyclic flexion-extension movement in groups I, II. The rabbits in group I and III were sacrified for the histological evaluation at postoperative fifth day. The rabbits in groups I II, IV and V were sacrified at postoperative 21st day. There was no peridural venous endothelial injury or stasis but there was an increased amount of polymorph nuclear leukocytes in both group I (unstable-overuse) and group III (unstable-no overuse) after sacrification at postoperative fifth day. Peridural fibrosis and also vascular changes with different grades were seen in group II, VI and V after sacrification at postoperative 21st day. The grade of the venous changes and the mean amount of peridural scar formation were prominently higher in group II (unstable-overuse) than in group IV (unstable-no overuse) and V (control group). There was no difference between group IV and V for peridural scar formation and vascular changes. In conclusion, the instability of the lumbar spine with overuse could be a cause of peridural venous circulatory impairment, resulting in fibrosis formation. PMID- 16382310 TI - Factors predicting outcome after whiplash injury in subjects pursuing litigation. AB - Records of 277 patients presenting for medicolegal reporting following isolated whiplash injury were studied retrospectively. A range of pre-accident, accident and response variables were recorded. Multivariate analysis was used to determine the main factors that predict physical and psychological outcome after whiplash injury. The factors that showed significant association with poor outcome on both physical and psychological outcome scales were pre-injury back pain, high frequency of General Practitioner attendance, evidence of pre-injury depression or anxiety symptoms, front position in the vehicle and pain radiating away from the neck after injury. The strongest associations were with factors that are present before impact. In this selected cohort of patients, there is a physical and a psychological vulnerability that may explain the widely varied response to low violence indirect neck injury. PMID- 16382311 TI - Gregarious behaviour of evasive prey. AB - Gregarious behavior of potential prey was explained by Hamilton (1971) on the basis of risk-sharing: The probability of being picked up by a predator is small when one makes part of a large aggregate of prey. This argument holds only if the predator chooses its victims at random. It is not the case for herds of evasive prey in the open, where prey's gregarious behavior, favorable for the fast group members, makes it easier for the predator to home in on the slowest ones. We show conditions under which gregarious behavior of the relatively fast prey individuals leaves slowest prey with no other choice but to join the group. Failing to do so would signal their vulnerability, making them a preferred target for the predator. Analysis of an n + 1 player game of a predator and n unequal prey individuals clarifies conditions for fully gregarious, partially gregarious, or solitary behavior of the prey. PMID- 16382312 TI - A stochastic model and a functional central limit theorem for information processing in large systems of neurons. AB - The paper deals with information transmission in large systems of neurons. We model the membrane potential in a single neuron belonging to a cell tissue by a non time-homogeneous Cox-Ingersoll-Ross type diffusion; in terms of its time varying expectation, this stochastic process can convey deterministic signals. We model the spike train emitted by this neuron as a Poisson point process compensated by the occupation time of the membrane potential process beyond the excitation threshold. In a large system of neurons 1 < or = i < or = N processing independently the same deterministic signal, we prove a functional central limit theorem for the pooled spike train collected from the N neurons. This pooled spike train allows to recover the deterministic signal, up to some shape transformation which is explicit. PMID- 16382313 TI - Geometric properties of a class of piecewise affine biological network models. AB - The purpose of this report is to investigate some dynamical properties common to several biological systems. A model is chosen, which consists of a system of piecewise affine differential equations. Such a model has been previously studied in the context of gene regulation and neural networks, as well as biochemical kinetics. Unlike most of these studies, nonuniform decay rates and several thresholds per variable are assumed, thus considering a more realistic model. This model is investigated with the aid of a geometric formalism. We first provide an analysis of a continuous-space, discrete-time dynamical system equivalent to the initial one, by the way of a transition map. This is similar to former studies. Especially, the analysis of periodic trajectories is carried out in the case of multiple thresholds, thus extending previous results, which all concerned the restricted case of binary systems. The piecewise affine structure of such models is then used to provide a partition of the phase space, in terms of explicit cells. Allowed transitions between these cells define a language on a finite alphabet. Some words are proved to be forbidden in this language, thus improving the knowledge on such systems in terms of symbolic dynamics. More precisely, we show that taking these forbidden words into account leads to a dynamical system with strictly lower topological entropy. This holds for a class of systems, characterized by the presence of a splitting box, with additional conditions. We conclude after an illustrative three-dimensional example. PMID- 16382314 TI - [Retinoblastoma]. AB - Retinoblastomas are the most frequent intraocular tumors in childhood. Untreated, the tumor is almost always fatal. Using a multidisciplinary approach combining the efforts of ophthalmologists, radiation oncologists, pediatric oncologists and geneticists a survival rate of more than 95% can be achieved. Molecular genetic research on the origin of retinoblastomas has substantially helped in our understanding of the origin of malignant tumors in general, as well as to the key role of the Rb-1 gene as a tumor suppressor. PMID- 16382315 TI - Transplantation of infant en bloc kidneys into paediatric recipients. AB - En bloc renal transplantation (EBT) from infant donors is an option for children with end-stage renal failure. Owing to potential complications, EBT is not performed in all paediatric nephrology centres. We evaluated the perioperative and long-term course of five children undergoing EBT. Primary diagnosis was atypical (diarrhoea-negative) haemolytic uraemic syndrome (n=2), interstitial nephropathy (two siblings) and branchio-oto-renal syndrome (n=1). Recipient and donor ages ranged between 5.9 and 11.1 years and 0.3 and 2.5 years, respectively. Follow-up time after EBT was 2.1-13.2 years. Perioperative complications included (1) a renal artery thrombosis, with immediate intraoperative reconstruction and primary non-functioning of the graft, with recovery after 10 days, and (2) a vesico-ureteric obstruction, successfully managed with temporary insertion of a JJ-catheter. All grafts had good long-term function. Absolute glomerular filtration rate (GFR; millilitres/minute) increased in all patients, whereas relative GFR (millilitres/minute per 1.73 m(2) body surface area) remained stable during the follow-up period in all but one. Kidney size increased significantly, with maximal growth during the first year after EBT; magnetic resonance imaging (MRI) showed normal structure and vasculature. EBT is a safe and effective option for young children with end-stage renal failure. Absolute GFR and graft size increase and adapt to the children's growing body mass. PMID- 16382316 TI - Clinical and molecular findings in a family with the carbonic anhydrase II deficiency syndrome. AB - Carbonic anhydrase II (CA2) deficiency syndrome is an autosomal recessive disorder leading to osteopetrosis, renal tubular acidosis, and cerebral calcifications. Affected members of an Arab family with the CA2 deficiency syndrome carried the "Egyptian mutation" in CA2, i.e., c.191 del A, H64fsX90. One affected member, homozygote for the mutation, developed primary pulmonary hypertension. Primary pulmonary hypertension was never described before in patients with this unique syndrome. The likelihood of both occurring randomly in a single individual is very low. We therefore speculate that there might be a possibility of an etiologic link between these entities. PMID- 16382317 TI - Changes in physical and psychosocial functioning among adolescents with chronic kidney disease. AB - Little research has been published assessing changes in the functional health status of children and adolescents with chronic kidney disease (CKD). We know little about which clinical parameters influence functional status or health related quality of life in these young people. In a prospective study using data from semi-annual visits over a 4-year period from 78 adolescents with CKD aged 11 years to 18 years, we detail the impact of several clinical measures (i.e., kidney function, albumin, hematocrit, height) on short-term changes in health related quality of life. The 50-item Child Health Questionnaire Parent Form, a validated health-related quality of life measure in children, was used to obtain physical and psychosocial functioning summary scores at each visit. After adjustment for the variables mentioned above, the physical summary score on the Child Health Questionnaire (CHQ) declined as glomerular filtration rate declined. Increasing height was associated with a positive change in physical and psychosocial summary scores. We conclude that decline in kidney function is associated with a subsequent decline in health-related quality of life, particularly in terms of physical activity. PMID- 16382318 TI - Compensatory renal growth due to neonatal ureteral obstruction: implications for clinical studies. AB - In response to unilateral ureteral obstruction (UUO), the contralateral kidney undergoes compensatory renal growth, which is enhanced in early development. We investigated the renal growth response to UUO in the neonatal rat. Within 2 days of birth, animals were subjected to sham-operation, complete UUO, or variable partial UUO, and kidneys were harvested 3-60 days later. Contralateral kidney weight increased after only 7 days of complete UUO. Increase in contralateral kidney weight was not significant for partial UUO until 45 days, but kidney/body weight ratio increased after only 14 days of 0.3 mm partial UUO. The rate of contralateral renal growth increased with age and with increasing severity of UUO. In rats subjected to 45 days UUO, glomerular area was proportional to kidney/body weight ratio (r =0.61, p <0.01). We conclude that the rate of compensatory renal growth is dependent on the severity and duration of obstruction, and takes place at the single nephron level. The results suggest that biologic variability limits the early detection of compensatory renal growth, which is compounded by limitations in measuring renal size by clinical imaging. Factoring kidney length (or volume) by intervertebral length (or body surface area) should improve the precision of tracking renal growth. PMID- 16382319 TI - Prophylactic calcium and vitamin D treatments in steroid-treated children with nephrotic syndrome. AB - Steroid treatment has several side effects, including the deterioration of the bone and mineral metabolism in children with nephrotic syndrome. This randomized prospective study was conducted to determine the effects and prophylactic role of calcium plus vitamin D treatment on bone and mineral metabolism in children receiving prednisolone treatment. 40 children (27 boys and 13 girls) with NS (18 new onset and 22 relapsing) were included in the study. Their mean age was 4.6+/ 1.8 years. All patients received prednisolone treatment (2 mg/kg/day for 4 weeks followed by alternate days at the same dose for 4 weeks). The patients were randomized into treatment (vitamin D 400 IU plus calcium 1 g daily) and non treatment groups. Bone mineral density, serum Ca, P, alkaline phosphatase and urinary Ca and P excretions were analyzed at the beginning and 2 months after the treatment. The XR36 Norland device was used for bone mineral density analysis. Bone mineral density was significantly decreased in both the treatment (0.54+/ 0.15 to 0.51+/-0.1 g/cm(2), P =0.001) and non-treatment (0.52+/-0.18 to 0.45+/ 0.16 g/cm(2), P <0.001) group. But the percentage of bone mineral density decrease was found to be significantly lower in the treatment group than in the non-treatment group (4.6+/-2.1% vs. 13.0+/-4.0%, respectively; P <0.001). Serum calcium and urinary calcium excretion increased in the treatment group (8.0+/-1.0 to 10.0+/-0.5 mg/dl and 1.1+/-0.5 to 3.2+/-1.0 mg/kg/day) and non-treatment group (8.1+/-0.8 to 10.0+/-0.6 mg/dl and 1.4+/-0.9 to 3.8+/-3.3 mg/kg/day) after prednisolone treatment (P <0.001). Steroid treatment decreases bone mineral density in children with nephrotic syndrome. Vitamin D plus calcium therapy at the current doses reduces but does not completely prevent bone loss, with no additional adverse effects. PMID- 16382320 TI - Laparoscopic adrenalectomy of an adrenal adenoma with myelolipoma relieves severe hypertension in a 16-year-old patient. AB - Adrenal adenoma with myelolipoma is extremely rare in pediatrics. Although the tumor is usually asymptomatic, sometimes it may result in serious manifestations. A 16-year-old patient was diagnosed with severe hypertension associated with a right adrenal mass. The laboratory work-up was inconclusive of the nature of the tumor. Plasma and urinary hormonal studies were not diagnostic. Magnetic resonance imaging (MRI) of the brain and meta-iodobenzylguanidine (MIBG) scanning were normal. MRI of the abdomen showed a heterogeneous adrenal mass 4.2x3.3 cm. Laparoscopic resection of the mass was done, and the pathology revealed an adrenal adenoma with myelolipoma. After tumor resection the hypertension resolved, and within 1 month the patient was off medications. At 2-year follow-up the patient's blood pressure remained normal at 120/73 mmHg. This is a case in which an adrenal adenoma with myelolipoma, a benign and usually asymptomatic tumor, presented as severe hypertension resolving with surgical resection of the tumor. PMID- 16382321 TI - Severe renovascular hypertension in an infant with congenital solitary pelvic kidney. AB - Renal artery stenosis (RAS) is one of the most common causes of severe arterial hypertension in infants. Its management is very difficult, especially when present in a single kidney. We report a case of severe hypertension caused by RAS of congenital single pelvic kidney in a 4-month-old boy. The patient presented with cardiorespiratory insufficiency that was first treated as acute fulminate myocarditis. Medical treatment of arterial hypertension was disappointing, as it had to be balanced between congestive cardiac failure and acute renal failure. Percutaneous transluminal angioplasty (PTA) done by coronary balloon dilatation catheters through the left axillary access was successful. Following dilatation of the renal artery, blood pressure decreased and its good control was possible by only one drug. With improved medical blood pressure control and normal growth development, the reassessment of clinical therapy options adjusted to a larger vessel size would be possible. Renovascular hypertension due to RAS in infants with a solitary kidney is difficult to control by medical treatment alone. PTA should be considered as a viable option in infants with refractory hypertension due to renal artery stenosis in a solitary kidney, since it has the potential of improving hypertension while preserving renal function. PMID- 16382322 TI - Hypoelectrolytemia due to inadequate diet. AB - Rice soup is a traditional Korean food administered to control diarrhea. Useful for short-term control of diarrhea, chronic consumption of this soup can cause problems. We treated an infant who visited our hospital for symptoms of lethargy due to hypoelectrolytemia, which developed from the continuous consumption of rice soup (the upper part of rice ground in water) mixed with a spoonful of milk over a 2-month period to control frequent diarrhea. The case is reported here, together with a brief review of the literature. PMID- 16382323 TI - Discordant evolution of nephrotic syndrome in mono- and dizygotic twins. AB - Twins represent a powerful resource for revealing multifactorial mechanisms in human diseases. Few reports are available on nephrotic syndrome in twins, and most furnish only a partial description of genetic identity based on human leukocyte antigens (HLA) analysis. We describe two pairs of mono and dizygotic twins with nephrotic syndrome who presented discordant outcomes in terms of length and required therapies. In one case, evolution to focal glomerulosclerosis was also documented. The basic molecular work-up included analysis of concordance based on 10 polymorphic markers (D3S1358, vVA, FGA, amelogenin, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820) and exclusion of the major slit-diaphragm gene mutation (NPHS2, CD2AP, WT1) causing nephrotic syndrome. To our knowledge, this is the first description of long-term outcome in mono- and dizygotic twins with proven genetic concordance. Discordant outcomes indicate a major influence of environmental and/or epigenetic multifactorial mechanisms on persistence and evolution of the disease to focal-segmental glomerulosclerosis. PMID- 16382324 TI - Urinary aquaporin-2 in children with acute pyelonephritis. AB - Children with acute pyelonephritis develop polyuria and have reduced maximum urinary concentration capacity. We studied whether these abnormalities are associated with altered urinary excretion of the water channel aquaporin-2 (AQP2) in the renal collecting duct. AQP2 is the main target for antidiuretic action of arginine vasopressin (AVP), and the urinary excretion of this protein is believed to be an index of AVP signaling activity in the kidney. Children with acute pyelonephritis, aged 5-14 years, were examined for urinary flow rate, creatinine clearance, unchallenged urine osmolality, and urinary ion excretion. Urinary excretion of AQP2 was measured by dot immunoblotting technique. Studies were performed in the acute phase of pyelonephritis, in the same children after treatment, and in control patients. At the onset of pyelonephritis, urinary flow rate and solute excretion were increased, but the urinary osmolality was unchanged. The urinary level and urinary excretion of AQP2 was increased in acute pyelonephritis and decreased after treatment. Excretion of aquaporin-3 was unchanged, suggesting that the increase in AQP2 urinary excretion was not due to a shedding of collecting duct cells. The results suggest that a mechanism proximal to the collecting duct may be responsible for the polyuria observed in children with acute pyelonephritis. Increased urinary AQP2 levels suggest that a compensatory activation of apical plasma membrane targeting of AQP2 may occur in pyelonephritis. PMID- 16382325 TI - In vitro analysis of the effect of hyperbilirubinemia on rabbit ureter and bladder. AB - Spontaneous resolution of intrauterine pelvic dilatations after birth is an expected outcome. In nonobstructive pelvic dilatations, changes in ureteral and bladder physiology may also play a part. We aimed to demonstrate the effect of increased concentrations of bilirubin on ureteral and bladder muscles in vitro. Normal and pathologic concentrations of bilirubin (3.5x10(-7)-10(-5)M and 10(-4) 4x10(-4)M, respectively) caused no change in the basal ureter tension (343.9+/ 29.4 mg). Normal concentrations of bilirubin caused no difference in basal bladder tension (430.2+/-70.2 mg), but pathologic concentrations caused a decrease of 303.8+/-52.9 mg. Normal and pathologic amounts of bilirubin were cumulatively applied to rabbit ureteral and bladder tissues both after reaching basal tension and when contracted with KCl (80 mM and 120 mM KCl for ureter and bladder, respectively). The cumulative addition of normal bilirubin concentrations to the ureteral tissues precontracted with KCl produced 86.4+/ 7.2% relaxation, while the addition of pathologic bilirubin concentrations produced a relaxation of 133.9+/-17.4%, which was significantly higher (p=0.04). Similarly, the addition of normal concentrations of bilirubin to the bladder tissues precontracted with KCl produced a maximal relaxation of 35.3+/-2.2%, while pathologic concentrations produced a maximal relaxation of 53.5+/-3.5%, which was significantly higher (0.001). Consequently, high concentrations of bilirubin caused a mild relaxation in basal ureteral and bladder tensions, while pathologically increased concentrations led to significant relaxation in both types of precontracted tissues. We suggest that high bilirubin levels may partly but not directly contribute to the spontaneous recovery of hydronephrosis because of the relaxation effect on bladder while probably causing susceptibility to urinary tract infections because of relaxation of both ureteral and bladder tissues. PMID- 16382326 TI - "Bartter-like" phenotype in Kearns-Sayre syndrome. AB - Kearns-Sayre syndrome (KSS) is a mitochondrial disease caused by large deletions in mitochondrial DNA (mtDNA). In most patients the disease is characterized by mtDNA heteroplasmy, where a mixture of wild-type and mutated mtDNA co-exist within cells in variable proportion, modulating the severity of the phenotype in different tissues. We report on the case of a 14-year-old child with classical symptoms of KSS and a renal phenotype characterized by hypokalaemic alkalosis, hypomagnesaemia, hyperreninaemia, hyperaldosteronism and nephrocalcinosis, resembling Bartter syndrome. Analysis of mtDNA demonstrated an 8,661 bp deletion involving eight mitochondrial genes. Uneven degrees of mtDNA heteroplasmy were demonstrated in several tissues, ranging from 24% to 60% of deleted/total mtDNA. Variable degrees of expression of mitochondrial enzymes were also found in biopsy specimens of renal and skeletal muscle by histocytochemistry. In particular, preserved cytochrome c oxidase was observed in tubular structures within medullary rays. It is proposed that a "Bartter-like" phenotype can arise in some patients with KSS as a result of heteroplasmy. In these cases aldosterone responsive tubular structures have been spared during renal embryogenesis, allowing for the development of hypokalaemic alkalosis in response to salt and water losses from the more damaged tubular segments. PMID- 16382327 TI - Continuous peritoneal dialysis in children: a single-centre experience in a developing country. AB - Continuous peritoneal dialysis (CPD) is the most commonly used modality of dialysis in children. Continuous ambulatory peritoneal dialysis (CAPD) has been an established form of therapy in adult patients with end-stage renal failure in India for more than a decade. There is a paucity of published experience of CPD in children from developing countries. We retrospectively studied children with end-stage renal failure (ESRD) that had been on CAPD over the past 10 years. Thirty patients with ESRD, mean age 13+/-8 years (range 5-21 years), male 18, were started on CAPD from 1994 to October 2004. The mean break-in period was 12+/ 3 days. Of these 30 patients, 15 had a total of 21 episodes of peritonitis. The peritonitis rate was 0.58 episodes per patient year. E. coli was the commonest organism causing peritonitis. On outcome analysis, 7/30 (23.3%) patients received a renal transplant, while 11/30 (36.6%) continued on CAPD, awaiting a kidney transplant. Of the rest, eight (26.6%) patients died, two (6.7%) suffered technique failure and were changed to haemodialysis, and two (6.7%) were lost to follow-up after 2 months. The mean cumulative survival time of patient on CPD was 42 months. We conclude that CPD is a viable option for dialysis in ESRD children in a developing country and is a successful bridge between ESRD and renal transplantation. PMID- 16382328 TI - Ultrasound appearance of tendon tears. Part 2: lower extremity and myotendinous tears. AB - Traumatic tears of the musculotendinous complex at the lower limb are common in clinical practice but can be difficult to detect and to evaluate because of swelling and pain that can limit proper physical examination. They can affect sedentary subjects or active sports participants involved in amateur or professional activities. In the first group tendons are more commonly affected, while myotendinous tears are common in sports players. The aims of this review article are to review the aetiology and pathomechanism of the most common ruptures affecting the tendons and the main myotendinous junctions of the lower extremity and to describe their ultrasound findings as well as to correlate ultrasound appearance with that of the other imaging modalities. PMID- 16382329 TI - Can spectro-temporal complexity explain the autistic pattern of performance on auditory tasks? AB - To test the hypothesis that level of neural complexity explain the relative level of performance and brain activity in autistic individuals, available behavioural, ERP and imaging findings related to the perception of increasingly complex auditory material under various processing tasks in autism were reviewed. Tasks involving simple material (pure tones) and/or low-level operations (detection, labelling, chord disembedding, detection of pitch changes) show a superior level of performance and shorter ERP latencies. In contrast, tasks involving spectrally and temporally-dynamic material and/or complex operations (evaluation, attention) are poorly performed by autistics, or generate inferior ERP activity or brain activation. Neural complexity required to perform auditory tasks may therefore explain pattern of performance and activation of autistic individuals during auditory tasks. PMID- 16382330 TI - Early postnatal care among healthy newborns in 19 States: pregnancy risk assessment monitoring system, 2000. AB - OBJECTIVE: To examine early postnatal care among healthy newborns during 2000 in 19 states. METHODS: Using data from the Pregnancy Risk Assessment Monitoring System, a multistate population-based postpartum survey of women, we calculated prevalences of early discharge (ED; stays of < or =2 days after vaginal delivery and < or =4 days after Cesarean delivery) and early follow-up (within 1 week) after ED. We used logistic regression to estimate adjusted odds ratios (aOR) and 95% confidence intervals (CI) describing how ED and lack of early follow-up were associated with state legislation and maternal characteristics. RESULTS: While most healthy term newborns (83.5-93.4%) were discharged early, and most early discharged newborns (51.5-88.5%) received recommended early follow-up, substantial proportions of early-discharged newborns did not. Compared with newborns in states where legislation covered both length of hospital stay (LOS) and follow-up, newborns in states without such legislation were more likely to have ED (aOR: 1.25; CI: 1.01-1.56). Lack of early follow-up was more likely among newborns in states with neither LOS nor follow-up legislation (aOR: 2.70, CI: 2.32-3.14), and only LOS legislation (aOR: 1.38, CI: 1.22-1.56) compared with those in states with legislation for both. ED was more likely among newborns born to multiparous women and those delivered by Cesarean section and less likely among those born to black and Hispanic mothers and mothers with less education. CONCLUSIONS: Lack of early follow-up among ED newborns remains a problem, particularly in states without relevant legislation. These findings indicate the need for continued monitoring and for programmatic and policy strategies to improve receipt of recommended care. PMID- 16382331 TI - A positive orientation toward early motherhood is associated with unintended pregnancy among New Orleans youth. AB - OBJECTIVE: Characterizing young women's willingness to enter motherhood is critical to understanding the high rates of unintended pregnancy among women under 20 years. Our objectives were to discuss a measure called Positive Orientation towards Early Motherhood (POEM), and investigate its association with self-reported unintended pregnancy experience. METHODS: We used data from 332 African-American women 13-19 years old recruited at public family planning and prenatal clinics in New Orleans. Using a series of ANOVAs and multinomial logistic regression, we assessed differences in POEM between four different outcome groups: women who were never pregnant and those who had only intended pregnancies, only unintended pregnancies and both unintended and intended pregnancies. RESULTS: The data suggested that young women perceive pregnancy as an opportunity to assert responsibility, become closer with their families and achieve greater intimacy with their boyfriends. Multiple regression analysis indicated that this positive orientation toward early motherhood independently raised the likelihood that young women experienced unintended pregnancies. In particular, the perception that a pregnancy makes a young woman feel more responsible was associated with an increased likelihood that a young woman had only unintended pregnancies compared to no pregnancies at all. Interestingly, this perception did not differentiate young women who had only intended pregnancies from those who were never pregnant. CONCLUSION: When interpreting reports of unintended pregnancy, more attention should be given to young women's orientation toward early motherhood. Doing so will inform policies that address both personal and structural factors that contribute to persistently high rates of unintended pregnancy among adolescents. PMID- 16382332 TI - Unmet need and problems accessing specialty medical and related services among children with special health care needs. AB - OBJECTIVES: To extend what is known about parent reports of their child's need for specialty medical and related services, unmet need, and specific types of access problems among children with special health care needs (CSHCN). METHODS: Using data from a 1998-1999 20-state survey of families of CSHCN, we examined differences in parent report of need for services by child characteristics, investigated parent report of unmet need and access problems by service area and number of services needed, and estimated the likelihood of four access problems and unmet need by child, family, and health insurance characteristics. RESULTS: Overall, the sample children had numerous service needs, although the prevalence of need varied by service type and child characteristics. Reports of unmet need were greater for older children and for children with multiple service needs, unstable health care needs or a behavioral health condition, parents who were in poor health or had more than a high school education, and families whose insurance coverage was inconsistent or lacked a secondary plan. Reports of access problems were greatest for mental health and home health services. The two most prevalent access problems were finding a skilled provider and getting enough visits. CONCLUSIONS: The results underscore the importance of finding new ways to link children with behavioral health problems to mental health services, implementing coordinated care and the other core dimensions of the medical home concept, increasing the number of specialty pediatricians and home health providers, and expanding coverage for a wider range of mental health services. PMID- 16382333 TI - The relationship of depressive symptoms to parenting competence and social support in inner-city mothers of young children. AB - OBJECTIVE: Despite the high prevalence of maternal depression and its negative consequences for children, many pediatricians fail to identify this problem. Our goal was to determine whether simple questions about parenting competence and the adequacy of maternal social support might be useful to providers in determining which inner-city mothers are likely to be depressed. METHODS: We surveyed a convenience sample of 279 English-speaking mothers of children 6 months to 3 years old prior to a routine visit at an urban, hospital-based general pediatrics clinic. The mothers self-completed the Psychiatric Symptom Index (PSI) and the Parenting Stress Index Sense of Competence subscale, and rated the adequacy of their social support, and provided health and sociodemographic data by face-to face interview. RESULTS: 41% of mothers had "high" PSI symptom levels and 22% had scores above a criterion that suggests major depressive disorder. In addition, 15% experienced high parenting stress (low competence) and 42% reported little or no social support. High distress was unrelated to a variety of sociodemographic risk factors, but significantly associated with a poor sense of parenting competence (Adj. OR = 3.3, 95% CI = 1.5, 7.0) and inadequate perceived social support (Adj. OR = 2.3, 95% CI = 1.2, 4.4), as well as with having health-related activity limitations (Adj. OR = 3.2, 95% CI = 1.1, 9.0). CONCLUSIONS: Negative ratings of parenting competence, low perceived social support, and presence of health-related activity restrictions can be useful markers of likely depression among inner-city mothers of young children. These factors are often assessed during routine pediatric visits and may be helpful to pediatricians in identifying mothers needing further evaluation or treatment by mental health specialists. PMID- 16382334 TI - Time-resolved fluorescence studies on bovine serum albumin denaturation process. AB - The denaturation of Bovine Serum Albumin (BSA) by a chaotropic agent, guanidinium hydrochloride (GuH+Cl-) was studied by fluorescence lifetime analysis. The BSA was labelled with 1-anilino-8-naphthalene sulfonate (ANS) at two different molar ratios (1:1) and (1:10). The non-exponential fluorescence kinetics of the BSA-ANS complex at different stages of denaturation is analysed using three different models: a discrete tri-exponential sum, stretched exponential, and Gaussian lifetime distribution. In all cases, the fluorescence decay times decreased with protein denaturation. The results from the models show that there are at least two different binding sites located in the BSA protein with different water accessibility. PMID- 16382336 TI - Endogenous neuro-protectants in ammonia toxicity in the central nervous system: facts and hypotheses. AB - The paper overviews experimental evidence suggestive of the engagement of three endogenous metabolites: taurine, kynurenic acid, and glutathione (GSH) in the protection of central nervous system (CNS) cells against ammonia toxicity. Intrastriatal administration of taurine via microdialysis probe attenuates ammonia-induced accumulation of extracellular cyclic guanosine monophosphate (cGMP) resulting from over-activation of the N-methyl-D: -aspartate/nitric oxide (NMDA/NO) pathway, and this effect involves agonistic effect of taurine on the GABA-A and glycine receptors. Taurine also counteracts generation of free radicals, increased release of dopamine, and its metabolism to dihydroxyphenylacetic acid (DOPAC). Taurine reduces ammonia-induced increase of cell volume (edema) in cerebrocortical slices by a mechanism involving GABA-A receptors. Massive release of radiolabeled or endogenous taurine from CNS tissues by ammonia in vivo and in vitro is thought to promote its neuroprotective action, by making the amino acid available for interaction with cell membranes and/or by driving excess water out of the CNS cells (astrocytes) that underwent ammonia induced swelling. Ammonia in vivo and in vitro affects in variable ways the synthesis of kynurenic acid (KYNA). Since KYNA is an endogenous NMDA receptor antagonist with a high affinity towards its glycine site, changes in its content may counter over-activation or depression of glutaminergic transmission observed at the different stages of hyperammonemia. GSH is a major antioxidant in the CNS whose synthesis is partly compartmented between neurons and astrocytes: astrocytic GSH is a source of precursors for the synthesis of neuronal GSH. Ammonia in vitro stimulates GSH synthesis in cultured astrocytes, which may compensate for increased GSH consumption (decreased GSH/GSSG ratio) in neurons. PMID- 16382338 TI - Limited capacity for ammonia removal by brain in chronic liver failure: potential role of nitric oxide. AB - Chronic liver failure leads to hyperammonemia and consequently increased brain ammonia concentrations, resulting in hepatic encephalopathy. When the liver fails to regulate ammonia concentrations, the brain, devoid of a urea cycle, relies solely on the amidation of glutamate to glutamine through glutamine synthetase, to efficiently clear ammonia. Surprisingly, under hyperammonemic conditions, the brain is not capable of increasing its capacity to remove ammonia, which even decreases in some regions of the brain. This non-induction of glutamine synthetase in astrocytes could result from possible limiting substrates or cofactors for the enzyme, or an indirect effect of ammonia on glutamine synthetase expression. In addition, there is evidence that nitration of the enzyme resulting from exposure to nitric oxide could also be implicated. The present review summarizes these possible factors involved in limiting the increase in capacity of glutamine synthetase in brain, in chronic liver failure. PMID- 16382337 TI - Molecular mechanisms of the alterations in NMDA receptor-dependent long-term potentiation in hyperammonemia. AB - Long-term potentiation (LTP) is a long-lasting enhancement of synaptic transmission efficacy and is considered the base for some forms of learning and memory. Hyperammonemia impairs LTP in hippocampus. Proper LTP induction in hippocampal slices requires activation of the soluble guanylate cyclase (sGC) protein kinase G (PKG)-cyclic guanosine monophosphate (cGMP)-degrading phosphodiesterase pathway. Hyperammonemia impairs LTP by impairing the tetanus induced activation of this pathway. The tetanus induces a rapid cGMP rise, reaching a maximum at 10 s, both in the absence or in the presence of ammonia. The increase in cGMP is followed, in control slices, by a sustained decrease in cGMP because of PKG-mediated activation of cGMP-degrading phosphodiesterase, which is required for maintenance of LTP. Hyperammonemia prevents completely tetanus-induced decrease in cGMP by impairing PKG-mediated activation of cGMP degrading phosphodiesterase. Addition of 8 Br-cGMP to slices treated with ammonia restores both phosphodiesterase activation and maintenance of LTP. Impairment of LTP in hyperammonemia may be involved in the impairment of the cognitive function in patients with hepatic encephalopathy. PMID- 16382339 TI - Protein tyrosine nitration in hyperammonemia and hepatic encephalopathy. AB - Hepatic encephalopathy is seen as a clinical manifestation of a chronic low grade cerebral edema, which is thought to trigger disturbances of astrocyte function, glioneuronal communication, and finally HE symptoms. In cultured astrocytes, hypoosmotic swelling triggers a rapid oxidative stress response, which involves the action of NADPH oxidase isoenzymes, followed by tyrosine nitration of distinct astrocytic proteins. Oxidative stress and protein tyrosine nitration (PTN) are also observed in response to ammonia, inflammatory cytokines, such as TNF-alpha or interferons, and benzodiazepines with affinity to the peripheral benzodiazepine receptor (PBR). NMDA receptor activation was identified as upstream event in protein tyrosine nitration (PTN). Cerebral PTN is also found in vivo after administration of ammonia, benzodiazepines or lipopolysaccharide and in portocaval shunted rats. PTN predominantly affects astrocytes surrounding cerebral vessels with potential impact on blood-brain-barrier permeability. Among the tyrosine-nitrated proteins, glutamine synthetase, GAPDH, extracellular signal regulated kinase and the PBR were identified. PTN of glutamine synthetase is associated with inactivation of the enzyme. Thus, factors known to trigger hepatic encephalopathy induce oxidative/nitrosative stress on astrocytes with protein modifications through PTN. The pathobiochemical relevance of astrocytic PTN for the development of HE symptoms remains to be established. PMID- 16382340 TI - Wilson disease. AB - Wilson disease (WD) is an autosomal recessive inherited disorder of copper metabolism, resulting in pathological accumulation of copper in many organs and tissues. The hallmarks of the disease are the presence of liver disease, neurologic symptoms, and Kayser-Fleischer corneal rings. The leading neurologic symptoms in WD are dysathria, dyspraxia, ataxia, and Parkinsonian-like extrapyramidal signs. Changes in the basal ganglia in brain magnetic resonance imaging (MRI) are characteristic features of the disease. In presence of liver cirrhosis, some features may resemble hepatic encephalopathy. Symptoms and MRI abnormalities may be fully reversible on treatment with zinc or copper chelators. Improvement can be monitored by serial recording of brain-stem-evoked responses. The basic defect is an impaired trafficking of copper in hepatocytes. ATP7B is the gene product of the WD gene located on chromosome 13 and resides in hepatocytes in the trans-Golgi network, transporting copper into the secretory pathway for incorporation into apoceruloplasmin and excretion into the bile. While about 40% of patients preset with neurologic symptoms, little is known about the role of copper and ATP7B in the central nervous system. In some brain areas, like in the pineal gland, ATP7B is expressed and functionally active. Increasing evidence supports an important role for metals in neurobiology. Two proteins related to neurodegeneration are copper-binding proteins (1) the amyloid precursor protein (APP), a protein related to Alzheimer's disease, and (2) the Prion protein, related to Creutzfeldt-Jakob disease. A major source of free radical production in the brain derives from copper. To prevent metal-mediated oxidative stress, cells have evolved complex metal transport systems. APP is a major regulator of neuronal copper homeostasis and has a copper-binding domain (CuBD). The surface location of this site, structural homology of CuBD to copper chaperones, and the role of APP in neuronal copper homeostasis are consistent with the CuBD acting as a neuronal metallotransporter. There are several copper containing enzymes in the brain, like dopamine beta hydroxylase or Cu/Zn superoxide dismutase (SOD1). Their function may be altered because of copper overload. WD appears to be associated with a dopaminergic deficit. Mutations in the SOD1gene cause familial amyotrophic lateral sclerosis. Survival of transgenic mice with a mutant SOD1 which fails to incorporate Cu((2+)) in its active site was improved by copper depletion. Wilson disease (WD) is an autosomal recessive inherited disorder in which copper pathologically accumulates primarily within the liver and subsequently in the neurologic system and many other organs and tissues. Presence of liver disease, neurologic symptoms, and Kayser-Fleischer corneal rings are the hallmarks of the disease. PMID- 16382342 TI - Role of phosphate-activated glutaminase in the pathogenesis of hepatic encephalopathy. AB - Disturbed body nitrogen homeostasis due to impaired hepatic urea synthesis leads to an alteration in inter-organ ammonia trafficking, resulting in hyperammonemia. Glutamine (Gln) synthase is the alternative pathway for ammonia detoxification. Gln taken up by several organs is split by the intramitochondrial phosphate activated enzyme glutaminase (PAG) into glutamate (Glu) and ammonia. In cirrhotic patients with portosystemic intrahepatic shunt, the main source of systemic hyperammonemia is the small intestine, and ammonia derives mainly from Gln deamidation. Recently, PAG has been found increased in cirrhotics showing minimal hepatic encephalopathy and, therefore, could be implicated in the production of systemic hyperammonemia in these patients. Intestinal PAG activity correlates with psychometric test and magnetic resonance spectroscopy findings. Moreover, nitric oxide and tumor necrosis factor seem to be the major factors regulating intestinal ammonia production in cirrhotics. In the brain, PAG localized into the astrocytes is responsible for ammonia and free-radical production. The blockade of PAG, using 6-oxo-5-norleucine, avoids the toxic effects of Gln accumulation in the brain. These data support an important role for intestinal and brain glutaminase in the pathogenesis of hepatic encephalopathy and could be a new target for future therapies. PMID- 16382343 TI - Which are the right cells to be used in a bioartificial liver? AB - Anno 2004 freshly isolated or cryopreserved porcine or human hepatocytes have been most frequently used as bio-component in clinically applied bioartificial livers (BALs). Phase 1 studies of all bio-component modalities showed safety, feasibility, and improvement of biochemical, neurological, and hemodynamic parameters. However, both the pilot-controlled clinical trial with C3A cells and the randomized larger clinical trial with cryopreserved porcine hepatocytes did not show significant improvement of survival by intention-to-treat analysis. Because of the xenotransplantation-related disadvantages of porcine cells and the shortage of primary human hepatocytes, other sources of bio-components have to be explored. The future lies in the development of one or more human hepatocyte cell lines, which will have minimal immunogenicity, no risk of xeno-zoonosis, and the requested functionality and availability. Primary sources for the development of such human cell lines are liver-tumor-derived cell lines, immortalized fetal or adult hepatocytes, and stem cells of hepatic, hematopoietic, or embryonic origin. At present the most promising results for BAL application have been obtained by immortalization of human fetal liver cells by reconstitution of telomerase activity. However, in all cell types tested so far, the in vitro differentiation cannot be stimulated to such an extent that their functionality reaches that of primary human hepatocytes. More insight in differentiation-promoting factors and the influence of matrix and co-culture conditions is needed. PMID- 16382341 TI - Mechanisms of ammonia-induced astrocyte swelling. AB - Astrocyte swelling represents the major factor responsible for the brain edema associated with fulminant hepatic failure (FHF). The edema may be of such magnitude as to increase intracranial pressure leading to brain herniation and death. Of the various agents implicated in the generation of astrocyte swelling, ammonia has had the greatest amount of experimental support. This article reviews mechanisms of ammonia neurotoxicity that contribute to astrocyte swelling. These include oxidative stress and the mitochondrial permeability transition (MPT). The involvement of glutamine in the production of cell swelling will be highlighted. Evidence will be provided that glutamine induces oxidative stress as well as the MPT, and that these events are critical in the development of astrocyte swelling in hyperammonemia. PMID- 16382344 TI - Neural synchronization in hepatic encephalopathy. AB - Hepatic encephalopathy (HE) is clinically characterized by a large variety of symptoms including motor symptoms, cognitive deficits, as well as changes in the level of alertness up to hepatic coma. A number of pathological processes affecting glial and neuronal function have been identified, including hyper ammonia, changes within the excitatory and inhibitory transmitter systems, as well as osmolytic changes with consecutive cell swelling. One explanation how these pathological processes result in neurological deficits in HE is the concept of pathologically synchronized oscillations within and between relevant brain regions. A number of studies suggest that the cognitive deficits and the reduced level of alertness in patients with HE can be attributed to a significantly slowed and pathologically synchronized spontaneous oscillatory brain activity, depending on the grade of HE. Moreover, HE motor symptoms, like postural tremor called"mini asterixis," have recently been shown to be associated with abnormal thalamo-cortical and cortico-muscular synchronization. Indirect evidence exists from studies of processing and recognition of flicker stimuli that in HE slowing of oscillations also occurs in the visual system. Taken together, pathological synchronization of neuronal activity may turn out to be a promising pathophysiological concept for linking neuronal dysfunction to the diversity of clinical deficits in HE. PMID- 16382345 TI - Altered modulation of motor activity by group I metabotropic glutamate receptors in the nucleus accumbens in hyperammonemic rats. AB - One of the neurological complications in hepatic encephalopathy is the impairment of motor coordination and function. Clinical signs of basal ganglia, cortico spinal and cerebellar dysfunction have been commonly detected in these patients. We are studying the molecular bases of the alterations in motor coordination and function in hepatic encephalopathy. Hyperammonemia is considered the main factor responsible for the neurological alterations in patients with hepatic encephalopathy. Activation of metabotropic glutamate receptors (mGluRs) in the nucleus accumbens (NAcc) induces locomotion in rats. Asa first step in our studies on the alterations in motor co-ordination and function in hyperammonemia and hepatic encephalopathy we studied whether the control of motor function by mGluRs in the NAcc is altered in hyperammonemic rats. The locomotor activity induced by injection into the nucleus accumbens (NAcc) of DHPG, an agonist of group I mGluRs was significantly increased in hyperammonemic rats. Injection of DHPG increased extracellular dopamine but not glutamate in the NAcc of control rats. In hyperammonemic rats DHPG-induced increase in dopamine was significantly reduced, and extracellular glutamate increased 6-fold. The content of mGluR 1 but not mGluR 5, is increased in the NAcc of hyperammonemic rats. Blockade of mGluR 1 completely prevented motor and neurochemical effects induced by DHPG. These results show that modulation of both motor function and extracellular concentration of neurotransmitters by mGluRs in the NAcc is altered in hyperammonemia. This may contribute to the alterations in motor function in hepatic encephalopathy. PMID- 16382346 TI - Attention, memory, and cognitive function in hepatic encephalopathy. AB - Deficits in attention and arousal play a major role in the clinical presentation of hepatic encephalopathy. Attention deficits are also the main components of minimal hepatic encephalopathy. The present paper summarizes some findings about attentional and memory dysfunction in hepatic encephalopathy, with reference to basic knowledge about normal attention and memory function and their cerebral representation. PMID- 16382347 TI - Neurophysiological investigation of hepatic encephalopathy. AB - The neurophysiological tools used to study hepatic encephalopathy (HE) are the electroencephalogram (EEG) and the evoked potentials (EPs), both exogenous and endogenous. These tools are used (1) to diagnose HE in patients with severe liver disease and mental alteration, (2) to grade overt HE and monitor the effect of treatment for HE, (3) to diagnose minimal HE (4) to predict the occurrence of episodes of overt HE or liver-related death. The rationale for the use of each of these tools together with their theoretical and practical role is reviewed. PMID- 16382348 TI - Impairment of response inhibition precedes motor alteration in the early stage of liver cirrhosis: a behavioral and electrophysiological study. AB - Abnormality in movement initiation may partially explain psychomotor delay of cirrhotic patients, even in the absence of overt hepatic encephalopathy (HE). Therefore, the aim of this study was to determine the mechanisms of psychomotor delay observed in patients with cirrhosis in the absence of overt HE. Fourteen patients with nonalcoholic cirrhosis and 12 healthy matched control subjects underwent the lateralized readiness potential (LRP) measurement elicited by a visuospatial compatibility task (Simon task). Stimulus-triggered LRPonset reflects the time in which response is selected, while response-triggered LRP onset reflects motor execution. Cirrhotic patients showed delayed reaction times (RTs) compared to controls, particularly those with trial-making test A (TMT-A) or electroencephalogram (EEG) alterations. Stimulus-triggered LRP onset was found to be delayed in cirrhotic patients compared to controls, with a significant Group-versus-Condition interaction, showing a reduced cognitive ability to cope with interfering codes, even in patients without minimal HE (MHE). Response triggered LRP was found to be delayed only in the patients with TMT-A or EEG alterations. In conclusion, cirrhotic patients without overt HE display a psychomotor slowing, depending first lyon response inhibition and only later accompanied by impaired motor execution. PMID- 16382349 TI - Chronic systemic endotoxin exposure: an animal model in experimental hepatic encephalopathy. AB - Plasma levels of gut-derived endotoxins (lipopolysaccharides, LPS) are often elevated in cirrhotics and are thought to contribute to hepatic encephalopathy. Circulating LPS activates macrophages to produce tumor necrosis factor alpha (TNF alpha) and other potentially cytotoxic proinflammatory mediators. A pathogenic role for endotoxins is supported by studies showing that treatment with Lacto bacillusor antibiotics, both of which reduce LPS-producing intestinal Gram negative bacteria, alleviates experimental liver damage. To mimic the "leaky gut" syndrome with endotoxin translocation into the circulation in cirrhotics, a new animal model was developed. Rats were chronically exposed to ethanol and for the four last weeks also infused with endotoxin into the jugular vein from subcutaneously implanted osmotic minipumps. Animals receiving endotoxin had elevated hepatic expression of both pro- and anti-inflammatory cytokines, but compared to ethanol treatment alone hepatic steatosis and inflammatory changes were only marginally increased. This demonstrates marked endotoxin tolerance, probably as a consequence of a counteracting anti-inflammatory cytokine response. The role of gut-derived endotoxin in hepatic encephalopathy has recently received considerable attention. To further delineate the role and actions of endotoxin and its extrahepatic effects, studies applying both acute challenge and chronic infusion seem warranted. The chronic endotoxin model, mimicking the "leaky gut," may best be combined with more robust ways to impair liver function, such as carbon tetrachloride treatment, bile duct ligation, or galactosamine administration. PMID- 16382350 TI - Animal models in the study of episodic hepatic encephalopathy in cirrhosis. AB - The availability of an animal model is crucial in studying the pathophysiological mechanisms of disease and to test possible therapies. Now, there are several models for the study of liver diseases, but there still remains a lack of a satisfactory animal model of chronic liver disease with hepatic encephalopathy (HE) and abnormalities in nitrogen metabolism, as seen in humans. In rats, two models of chronic HE are widely used: rats after portacaval anastomosis (PCA) and rats with chronic hyperammonemia. The first one mimics the situation induced in cirrhosis by collateral circulation, and has the problem of the absence of hepatocellular injury. The model of hyperammonemia is useful to study the effect of ammonia as a brain toxic substance, but also lacks liver failure. Bile-duct ligation has been used to induce cirrhosis and could also be a model of HE, probably with the addition of a precipitant factor. An ideal model of HE in chronic liver disease must have liver cirrhosis and a precipitant factor of HE; it must also show neuropathological characteristic findings of HE, neurochemical alterations in the main pathways impaired in these complications of cirrhosis, and low-grade brain edema. PMID- 16382351 TI - Acute liver failure: a critical appraisal of available animal models. AB - The availability of adequate experimental models of acute liver failure (ALF) is of prime importance to provide a better understanding of this condition and allow the development and testing of new therapeutic approaches for patients with ALF. However, the numerous etiologies and complications of ALF contribute to the complexity of this condition and render the development of an ideal experimental model of ALF more difficult than expected. Instead, a number of different models that may be used for the study of specific aspects of ALF have been developed. The most common approaches used to induce ALFin experimental animals are surgical procedures, toxic liver injury,or a combination of both. Despite the high prevalence of viral hepatitis worldwide, very few satisfactory viral models of ALF are available. Established and newly developed models of ALF are reviewed. PMID- 16382352 TI - Role of endogenous benzodiazepine ligands and their GABA-A--associated receptors in hepatic encephalopathy. AB - Benzodiazepine receptor ligands are suggested to play a role in the pathogenesis of hepatic encephalopathy (HE). Accumulation of these ligands in brain was suggested to explain in part the notion of"increased GABAergic tone," the rational for which arose initially from reports of a beneficial effect of the selective benzodiazepine antagonist flumazenil in HE patients. It was suggested on the basis of the effect of flumazenil in human HE that liver failure may result in alterations of the density and/or affinity of the benzodiazepine associated GABA-A receptor site. Subsequent controlled-clinical trials showed that fumazenil had a transient beneficial effect in only a subpopulation of HE patients. In contrast to the antagonists, partial inverse agonists of the benzodiazepine receptor have unequivocal beneficial effects on behavioral and electro-physiological performance in all experimental models of HE studied so far. Benzodiazepine-associated GABA-A receptors have consistently been demonstrated to be unaltered in both human and experimental HE. Contrary to initial reports, the so-called "endogenous benzodiazepines" do not appear to be significantly related to the pathogenesis of HE. On the other hand, nonbenzodiazepine GABA-A receptor complex modulators, such as neuro-steroids, recently identified in brain in human and experimental HE, may provide a new mechanistic basis for this disorder and lead to novel treatments for human HE. PMID- 16382353 TI - Racial bias in mock juror decision-making: a meta-analytic review of defendant treatment. AB - Common wisdom seems to suggest that racial bias, defined as disparate treatment of minority defendants, exists in jury decision-making, with Black defendants being treated more harshly by jurors than White defendants. The empirical research, however, is inconsistent--some studies show racial bias while others do not. Two previous meta-analyses have found conflicting results regarding the existence of racial bias in juror decision-making (Mazzella & Feingold, 1994, Journal of Applied Social Psychology, 24, 1315-1344; Sweeney & Haney, 1992, Behavioral Sciences and the Law, 10, 179-195). This research takes a meta analytic approach to further investigate the inconsistencies within the empirical literature on racial bias in juror decision-making by defining racial bias as disparate treatment of racial out-groups (rather than focusing upon the minority group alone). Our results suggest that a small, yet significant, effect of racial bias in decision-making is present across studies, but that the effect becomes more pronounced when certain moderators are considered. The state of the research will be discussed in light of these findings. PMID- 16382354 TI - Factors that facilitate and undermine children's beliefs about truth telling. AB - This study examined the extent to which children believe that truth telling is compromised by negative outcome expectancies. It also investigated the efficacy of two types of appeals, externally and internally directed, for encouraging truth telling. Seventy-two children from three age groups (5, 7, and 10 years of age) participated in a vignette study designed to examine these issues. Results showed that children believed that truth telling about an adult's transgression would be more likely if negative outcomes were not expected than if they were expected. Further, children believed that either externally or internally focused encouragement would facilitate truth telling when negative outcomes were expected for truth telling. Beliefs about the propensity for truth telling were associated more with positive evaluations of truth telling than with negative evaluations of lying. These results have important implications for court cases in which children testify about an adult who has sworn them to secrecy and they are afraid to speak the truth. PMID- 16382355 TI - The role of victim-related factors in victim restitution: a multi-method analysis of restitution in Pennsylvania. AB - Mandatory statutes do not always produce change, but a 1995 Pennsylvania statutory change making restitution mandatory dramatically increased the proportion of cases in which restitution was imposed. There are three possible reasons for this generally successful implementation: (a) judges agreed with the victim-centered goals of the statute, (b) there were mechanisms in place to implement the goals of assisting victims, and (c) there was a context supportive of victims that made it easier to follow the law. Two studies investigated these possible explanations. First, a statewide survey of trial court judges suggested that they agreed with the statute's goals of compensating victims. Second, hierarchical logistic models of 55,119 statewide restitution-eligible decisions indicated that a victim-related contextual factor, the nature and location of the victim/witness assistance office, was significantly related to the imposition of restitution, although a more general contextual factor relating to funding for victim programs had only small effects. PMID- 16382356 TI - Behavioral cues to deception vs. topic incriminating potential in criminal confessions. AB - Coding statements of criminal suspects facilitated tests of four hypotheses about differences between behavioral cues to deception and the incriminating potential (IP) of the topic. Information from criminal investigations corroborated the veracity of 337 brief utterances from 28 videotaped confessions. A four-point rating of topic IP measured the degree of potential threat per utterance. Cues discriminating true vs. false comprised word/phrase repeats, speech disfluency spikes, nonverbal overdone, and protracted headshaking. Non-lexical sounds discriminated true vs. false in the reverse direction. Cues that distinguished IP only comprised speech speed, gesticulation amount, nonverbal animation level, soft weak vocal and "I (or we) just" qualifier. Adding "I don't know" to an answer discriminated both IP and true vs. false. The results supported hypothesis about differentiating deception cues from incriminating potential cues in high stakes interviews, and suggested that extensive research on distinctions between stress-related cues and cues to deception would improve deception detection. PMID- 16382357 TI - The last word in court--a hidden disadvantage for the defense. AB - In the legal systems of most western countries, defense attorneys present their sentencing recommendation after the prosecution has presented its sentencing demands. This procedural sequence for criminal cases is intended to balance the impact of both parties on the judge's final decision. Especially the positioning of the defense's plea at the end of the trial follows the fundamental legal principle "in dubio pro reo." Research on judgmental anchoring, however, suggests that the standard procedural sequence may in fact work against this principle. Consistent with this implication, the present studies demonstrate that the defense's sentencing recommendation is anchored on, and consequently assimilated toward, the preceding recommendation by the prosecution. This influence prevents the defense attorney from effectively counterbalancing the prosecutor's demand. Instead, the biased defense attorney's recommendation partially mediates the impact of the prosecutor's demand on the judge's decision. These findings suggest that the standard procedural sequence in court may place the defense at a distinct disadvantage. PMID- 16382358 TI - Competence to waive interrogation rights and adjudicative competence in adolescent defendants: cognitive development, attorney contact, and psychological symptoms. AB - Although there is growing evidence of developmental differences in competency to waive interrogation rights and adjudicative competence, the correlates of adolescents' legal capacities remain unclear. This study examined the relationship of legal capacities to cognitive development, legal learning opportunities, and psychological symptoms. Participants were 152 male and female defendants aged 11-17, who completed Grisso's Instruments for Assessing Understanding and Appreciation of Miranda Rights, the Fitness Interview Test (Revised Edition), the Woodcock-Johnson III Cognitive Assessment Battery, and the Brief Psychiatric Rating Scale for Children. Legal capacities relevant to interrogation and adjudication increased with age. These developmental differences were partially mediated or explained by cognitive development. Of the specific cognitive abilities examined (general intellectual ability, verbal ability, reasoning, long-term retrieval, attention, and executive functioning), verbal ability was a particularly strong predictor of performance on competency measures. Also, defendants obtained lower scores on competency measures if they showed evidence of attention deficits or hyperactivity, had spent limited time with their attorneys, and/or were from low socioeconomic backgrounds. PMID- 16382359 TI - Interviewing former 1- and 2-year olds about medical emergencies 5 years later. AB - Five years earlier, 1- and 2-year-old children who had been injured seriously enough to require hospital ER treatment had been recruited from the ER. For this study, as many of these children as could be found participated. The majority of former 1-year olds recalled nothing about these highly stressful events, whereas most former 2-year olds recalled a great deal. For those former 1-year olds who did recall the target events, quality of recall was problematic. In particular, they made considerable source confusions or intrusions into their accounts of details from other related events, producing an account that amalgamated various events into one recollection. Forensic implications are discussed. PMID- 16382362 TI - Genetic evaluation of in situ conserved and reintroduced populations of wild rice (Oryza rufipogon: Poaceae) in China. AB - We evaluated the genetic consequences and efficiency of conservation practices in Oryza rufipogon using microsatellite DNA markers. Spatial autocorrelation analysis from 12 microsatellite loci revealed that microsatellite alleles were exclusively distributed in patches within the population, indicating that large populations were unlikely to be homogeneous. An in situ conserved stand of O. rufipogon, which has been protected by a concrete wall from a large population, captured only 67.9% of the total genetic variation of the previous large population. The concrete wall was built to protect the wild rice, but it acted more as a physical barrier to gene exchanges between the two sides. An assignment test revealed only 11.1% putative seed exchanges across the wall. A reintroduced population was found to be genetically very diverse. About 76.3% of the total genetic variation detected in other populations was captured in this reintroduced population, and 24.8% of the total genetic variation in this population was not found in other populations. These results display two important findings for conservation of O. rufipogon. First, conserving one part of a large population of O. rufipogon will not preserve an adequate sample of the genetic variability, since populations are not homogeneous, and genotype distribution varies among localities. Second, a reintroduced population is not genetically depauperate, but it is too early to assess its long-term survival. PMID- 16382363 TI - Biochemical genetic relationships among Tunisian hares (Lepus sp.), South African cape hares (L. capensis), and European brown hares (L. europaeus). AB - Tunisian hares (n = 45), currently assigned to Lepus capensis, were assayed for allelic variation at 40 allozyme loci, and allele frequencies at 32 loci were directly compared with earlier data of South African cape hares (L. capensis, n = 9) and European brown hares (L. europaeus, n = 244) to reveal genetic relationships among them. European mountain hares (L. timidus, n = 200) were used for outgroup comparison. In the Tunisian hares 27.5% of the loci were polymorphic with 2-4 alleles. Among all alleles at polymorphic loci, 15.1% occurred exclusively in Tunisian hares, 5.7% exclusively in cape hares, and 7.5% exclusively in brown hares at low frequencies. Not a single locus showed alternately fixed alleles between the samples of the L. capensis/L. europaeus complex. Levels of absolute and relative genetic differentiation among the samples of the L. capensis/L. europaeus complex were low, relative to pairwise comparisons involving mountain hares. Diverse cluster analyses and multidimensional scaling of various pairwise genetic distance matrices concordantly grouped Tunisian hares with brown hares, and South African cape hares clustered only slightly farther apart, whereas mountain hares were distinctly separate. These results suggest regionally distinct phylogenetic units within an overall cohesive gene pool in the L. capensis/L. europaeus complex, supporting Petter's view that all North African hares belong to L. capensis except for one local population of savanna hares, and that cape hares and brown hares are conspecific. PMID- 16382364 TI - A new variant of the mouse Akp1 locus. AB - A new electrophoretic migration type of alkaline phosphatase 1 (Akp1) was found on the cellulose acetate electrophoresis for kidney and liver homogenates of KWHM mouse, a newly established inbred strain derived from the Korean wild mouse (Mus musculus molossinus). This new type of alkaline phosphatase 1 was distinguished from previously reported AKP1A and AKP1B types in the mouse, and tentatively named AKP1C. In genetic analysis by mating experiments between KWHM and C57BL/6J (AKP1A) or BALB/cA (AKP1B), the phenotypic segregation ratios of AKP1A : AKP1AC : AKP1C or AKP1B : AKP1BC : AKP1C were 1 : 2 : 1 in both groups of F2 generations. It was therefore concluded that AKP1C type is controlled by Akp1c allele which is codominant with Akp1a and Akp1b alleles. PMID- 16382365 TI - Phylogenetic analysis of Bambusa (Poaceae: Bambusoideae) based on internal transcribed spacer sequences of nuclear ribosomal DNA. AB - Phylogenetic analyses of Bambusa species were performed using internal transcribed spacer sequences of nuclear ribosomal DNA. The 21 species sampled included members of Bambusa (sensu stricto), Dendrocalamopsis, Dendrocalamus, Guadua, Leleba, and Lingnania. Arundinaria gigantea was used as an outgroup. Using the maximum parsimony method with PAUP*, gaps were treated as missing states or new states. Parsimonious analysis revealed that Dendrocalamus latiflorus was closely related to the members of Dendrocalamopsis. Dendrocalamus membranaceus was a sister species to Dendrocalamus strictus. Three Dendrocalamus species were closely related to and nested in a polyphyletic Bambusa. Bambusa subaequalis was a sister species to B. multiplex, B. emeiensis to B. chungii, B. contracta to B. hainanensis, and B. flexuosa was a sister species to B. sinospinosa, B. tuldoides, B. surrecta, B. intermedia, and B. valida group, which raised doubts about the monophyly of the subgenera Bambusa (sensu stricto), Dendrocalamopsis, Leleba, and Lingnania under the genus Bambusa. PMID- 16382366 TI - A comparative study of genetic diversity of peripheral and central populations of chukar partridge from Northwestern China. AB - Although it has long been presumed that peripheral populations tend to exhibit low genetic diversity because of isolation and genetic drift, results of empirical investigation remain ambiguous. Some chukar partridge (Alectoris chukar) populations have expanded their ranges, resulting in several peripheral populations, due to recent deforestation by human beings in the Longdong Loess Plateau of northwestern China. On the basis of mitochondrial DNA control-region data, we compare the genetic diversity of two peripheral populations, Honghui and Wangxia, and six central populations. The Wangxia population possessed high levels of genetic diversity. The Honghui population, however, exhibited low genetic variation. The degree of isolation was the primary factor affecting the genetic diversity of the two peripheral populations. A peripheral population that was not isolated exhibited higher genetic diversity than did an isolated peripheral population. PMID- 16382367 TI - A new SNP in the 3'-UTR of the hsp 70-1 gene in Bos taurus and Bos indicus. PMID- 16382368 TI - Analysis of genetic polymorphism of deoxyribonuclease I in Ovambo and Turk populations using a genotyping method. AB - Deoxyribonuclease I (DNase I) polymorphism has been used as a valuable marker in genetic and clinical investigations. Six codominant alleles are known for DNase I, DNASE1*1, *2, *3, *4, and the recently discovered alleles *5 and *6. To detect these two new alleles, we added a new DNase I genotyping method based on both an allele-specific amplification and mismatched polymerase chain reaction (PCR). These methods were used to examine the distribution of DNase I genotypes in unrelated individuals from bloodstains of Ovambo and Turkish populations. The DNASE1*1 allele was found to be most dominant in the Ovambos. In contrast, Turks showed the highest allele frequency for DNASE1*2. This study is the first to demonstrate that there is a certain genetic heterogeneity in the worldwide distribution of DNase I polymorphism using the genotyping method of human DNase I polymorphism with PCR. PMID- 16382369 TI - Phospholamban gene mutations are not associated with hypertrophic cardiomyopathy in a Northern Greek population. AB - Hypertrophic cardiomyopathy (HCM) is a genetically transmitted cardiac disease characterized by unexplained myocardial hypertrophy and diverse clinical spectrum. Currently, more than 250 HCM-related mutations in 10 genes encoding contractile sarcomeric proteins have been identified. Phospholamban (PLN) is a modest modulator of intracellular Ca2+ homeostasis and may be a candidate gene responsible for cardiomyopathy. In this study 53 consecutive patients with HCM, coming from Northern Greece, were screened for mutations of PLN gene. The patients were evaluated by clinical history, physical examination, electrocardiogram and echocardiography. All PCR products were analyzed for mutation by both restriction analysis and sequencing. The systematic mutation screening did not reveal any mutation in exons 1 and 2 or in the promoter region of phospholamban gene. Additionally, no polymorphisms were detected in all patients. Therefore, PLN gene mutations were not found to be associated with HCM in a Northern Greece population. PMID- 16382371 TI - Breast reduction with inferior pedicle fascial suspension. AB - The ultimate aim of breast reduction surgery is to reduce breast tissue with long term maintenance of good breast shape. A technique using interwoven fascial flap suspension of the inferior pedicle through the pectoralis muscle and fascia is described. For this study, 25 patients were followed for 1 year after surgery. The vertical length of the breast between the lower midline of the areola and the new inframammary fold was measured 10 days after surgery and compared with a follow-up measurement 1 year later. These measurements were expressed as a ratio and compared with visual impressions. A ratio of more than 1.3 or a 2-cm difference in length or more was considered representative of breast "bottoming out." This was the case with two patients, both of whom had put on substantial weight after the procedure. The author contends that a consistently reliable technique that maintains good breast shape with more extensive scarring is preferable to a technique that occasionally "gets it right" with less scarring. Good scar management and variations in technique such as the fascial flap suspension described in this report have been of considerable benefit to the author's patients undergoing breast reduction surgery. PMID- 16382372 TI - Visible helical rim deformity resulting from excessive resection of postauricular skin during otoplasty. AB - An unusual deformity resulting from overresection of postauricular skin during otoplasty is presented. To the author's knowledge, this type of complication after otoplasty has not been described previously. PMID- 16382373 TI - Botulinum toxin treatment for oropharyngeal dysphagia due to tetanus. PMID- 16382375 TI - Metaanalyses - highest level of empirical evidence? PMID- 16382374 TI - Dietary and cancer-related behaviors of vitamin/mineral dietary supplement users in a large cohort of French women. AB - BACKGROUND: Several epidemiological studies suggested an association between vitamin/mineral dietary supplement use and cancer risk. However, characteristics of supplement users may themselves be related to cancer risk, and therefore could confound such etiological studies. Very little is known about the characteristics of French supplement users. AIM OF THE STUDY: To identify cancer-related behaviors and dietary characteristics of vitamin/mineral supplement users in the E3N cohort of French women. METHODS: Data on supplement use and cancer-related and socio-demographic characteristics were collected by self-administered questionnaires completed by 83,058 women, 67,229 of whom also completed a food frequency questionnaire. Supplement users were compared to non-users by unconditional logistic regression. RESULTS: Vitamin/mineral supplement users were significantly older and leaner (odds ratio [OR] for BMI >or= 30 vs. <18.5 kg/m2 = 0.35, 95% confidence interval [CI] 0.31-0.39), were less often current smokers, had a higher level of education and had more leisure physical activity. They used more phytooestrogen supplements (OR=3.95, 95 % CI 3.69-4.23), had more often a family history of breast cancer and had more often undergone cancer-screening. Users tended to have a healthier diet: less alcohol, more vegetables, fruit, dairy products, fish and soups. They had higher dietary intakes for most micro nutrients, fiber and omega3 fatty acids, lower fat intake and either similar or lower prevalence of inadequate dietary intake for all relevant nutrients except magnesium. CONCLUSIONS: To avoid major confounding, the lifestyle characteristics of supplement users should be considered in studies investigating the association between supplement use and cancer risk. PMID- 16382376 TI - Are the new antipsychotics no better than the classical neuroleptics? The problematic answer from the CATIE study. PMID- 16382377 TI - Proteomic analysis of the CSF in unmedicated patients with major depressive disorder reveals alterations in suicide attempters. AB - This is the first report on proteomic analysis of the cerebrospinal fluid (CSF) in unmedicated suicide attempters and non-attempters with major depressive disorder. Two-dimensional (2D) gel electrophoresis revealed that suicide attempters differed from non-attempters in one protein with an approximate molecular weight of 33 kD and an isoelectric point of 5.2. Proteomic analysis of the CSF is a promising non hypothesis-driven screening method for the detection of new candidate genes in neurobiological suicide research. PMID- 16382379 TI - Plasma levels of NT-pro-BNP in patients with atrial fibrillation before and after electrical cardioversion. AB - OBJECTIVE: Plasma levels of brain natriuretic peptide (BNP) have been examined in studies on patients with persistent atrial fibrillation, both before and after electrical cardioversion. Studied patients often showed a comorbidity with congestive heart failure, which complicates interpretation of measured BNP values as a natriuretic peptide. The aim of this study was to examine plasma levels of N terminal fragment pro-brain natriuretic peptide (NT-pro-BNP), which is the more stable but inactive cleavage product of pro-BNP in patients with atrial fibrillation, but normal left ventricular ejection fraction, before and after electrical cardioversion. PATIENTS AND METHODS: NT-pro-BNP plasma levels of 34 consecutive patients were measured before, shortly after and 11 days after electrical cardioversion. All patients showed a normal ejection fraction after echocardiographic or laevocardiographic criteria. RESULTS: At baseline, all patients showed elevated NT-pro-BNP compared to a healthy control group (1086 vs. 66.9 pg/ml, p<0.001). After a mean follow-up time of 11 days in patients with persistent restored sinusrhythm, NT-pro-BNP decreased from 1071 pg/ml at baseline to 300 pg/ml (p<0.001). In contrast, patients with recurrence of atrial fibrillation showed increased levels from 1570.5 pg/ml at baseline to 1991 pg/ml (p=0.13; n.s.). Recurrence of atrial fibrillation was independent from height of NT-pro-BNP levels at baseline (p=0.23). CONCLUSIONS: Atrial fibrillation in patients with a normal left ventricular ejection fraction is associated with elevated NT-pro-BNP plasma levels, which decrease when a persistent sinus-rhythm can be restored by electrical cardioversion. On the other hand, NT-pro-BNP seems to increase (n.s.) when recurrence of atrial fibrillation occurs. Finally, NT-pro BNP is no valid predictor for long-term success of sinus-rhythm restoration by electrical cardioversion. PMID- 16382380 TI - Experience with INR self-management: patient selection and complication rates. AB - BACKGROUND: INR self-management can reduce the risk of complications for patients with indication of long-term oral anticoagulation therapy. However, little is known about early indicators of complication risk. METHODS: In a prospective study on 330 consecutive patients all participants were informed about oral anticoagulation by a structured teaching program. The two groups were divided as to whether they received usual medical care provided by a family physician (n=220) or self-management (n=110) on a portable coagulation monitor (CoaguChek System). After a mean follow up of 13.3+/-4.4 months, the participants of the study were interviewed by a structured questionnaire to obtain information about hemorrhagic and thromboembolic complications as well as survival. RESULTS: In comparison to patients under usual care, patients with INR selfmanagement were significantly younger (58 vs 64 years) and had fewer comorbidities (diabetes and hypertension) as well as a higher ejection fraction (53.6 vs 51.1%). Indication for anticoagulation, age and heart rhythm explained 58% of the differences between the management groups. There was no significant difference in the overall complication rates between the two groups (usual care vs selfmanagement): major bleeding 0.5 vs 0.9%, cerebral embolism (TIA or stroke) 1.9 vs 0.9%, hospital admission 2.3 vs 1.8%. A high BMI (OR=1.5; 95% CI 1.06-1.25; p=0.001) or a high therapeutic INR range (OR=2.42; 95% CI 1.16-5.1; p=0.019) is associated with a higher complication rate. CONCLUSIONS: Complication rates for patients with long term oral anticoagulant therapy did not differ significantly between usual care and self-management. Rather, the patient's body weight and the requirement of high anticoagulation intensity drive the complication risk under both management systems. PMID- 16382381 TI - Slow pathway ablation in children with documented reentrant supraventricular tachycardia not inducible during invasive electrophysiologic study. AB - Radiofrequency catheter ablation (RFA) has become the procedure of choice for permanent therapy of atrioventricular nodal reentrant tachycardia (AVNRT). This report presents our experience with atrio-ventricular node (AVN) modification in patients with documented narrow complex reentrant SVT, but no evidence for an accessory pathway, and no inducible tachyarrhythmia during invasive electrophysiology (EP) study. METHODS: The study population consists of nine children, age range 6-13 years (median 9) with previously documented SVT who had no tachyarrhythmia inducible during EP study (at baseline and following isoprenaline infusion). Eight of the 9 EP studies were performed under general anesthesia, and one under conscious sedation. An accessory pathway was excluded in all patients by appropriate atrial and ventricular extrastimulus pacing techniques. Eight of the nine patients had dual AV nodal physiology, and one had single AV nodal echo beats. The slow AV nodal pathway was empirically ablated, by applying RF lesions in the right inferoseptal AV groove, achieving catheter tip temperature of 50 degrees C. The appearance of an accelerated junctional rhythm during RF application was deemed to denote a successful application site. AV conduction during RF application was confirmed by incremental atrial pacing. The catheter position, and its relation to the compact AV node was constantly monitored using the LocaLisa navigation system. The end-point was absence of dual AVN physiology, and/or AV nodal echo beats. RESULTS: Successful slow pathway ablation was achieved in all patients. One patient appeared to have two separate slow pathways with different locations and two AH-jumps, which were both successfully ablated. None of the patients had evidence of temporary or permanent AV block at follow-up (median duration 9 months, range 4 to 36 months); none has had recurrence of symptoms or documented tachyarrhythmia. CONCLUSIONS: In children with structurally normal hearts, a previously documented SVT, absence of an accessory pathway and noninducibility of SVT during EP study, empirical slow pathway ablation appears to be justified. PMID- 16382382 TI - Coronary fistula of right coronary artery to vena cava superior and ectasia of pulmonary artery. AB - We report on a patient with coronary heart disease, a coronary fistula of right coronary artery to vena cava superior and pulmonary hypertension. Combined with coronary artery revascularization, the coronary fistula was closed successfully. PMID- 16382383 TI - Successful treatment of oleander intoxication (cardiac glycosides) with digoxin specific Fab antibody fragments in a 7-year-old child: case report and review of literature. AB - A 7-year-old girl presented six hours after ingestion of a yellow oleander seed (Thevetia peruviana) with severe emesis, change in colour vision and complete heart block. Initial treatment with phenytoin and isoprenalin infusion led to intermittent supraventricular and ventricular tachycardia. The patient was then treated with two intravenous doses of 190 mg of digoxin-specific Fab antibody fragments (Digibind). Subsequently the patient's rhythm reverted to sinus rhythm and the symptoms resolved within 2 hours. CONCLUSION: administration of digoxin specific Fab antibody fragments in an otherwise healthy child after oleander intoxication is safe and without adverse reactions. PMID- 16382384 TI - Cardiac angiosarcoma: case report and review of the literature. AB - Angiosarcoma of the heart, the most common primary malignant cardiac tumor in adults is known to carry a dismal prognosis. The diagnosis is often delayed because of the nonspecific clinical presentation. Symptoms are determined by the size and location of the tumor. Echocardiography has become the primary diagnostic technique because of its high degree of accuracy, noninvasiveness, and cost effectiveness. Complete surgical resection is required for improved survival. Conventional postoperative chemotherapy does not appear to modify the clinical course. We report a case of cardiac angiosarcoma with a large mural mass infiltrating the right atrial and ventricular walls and critically review the pertinent literature. PMID- 16382385 TI - Pregnancy-associated spontaneous coronary artery dissection: impact of medical treatment. Case report and systematic review. AB - We report on a 22- year-old woman with postpartum dissection of the left anterior descending artery and the intermediate branch. The patient was treated with acetylsalicylic acid (ASA), clopidogrel, and betablocker only. Coronary angiography performed 20 months later revealed complete resolution of the dissection sites. The patient's cardiovascular risk factors included mild smoking and high total cholesterol and low-density-lipoprotein-cholesterol levels, which showed a marked fall after pregnancy without pharmacological cholesterol modifying therapy raising the question whether pregnancy-related hypercholesterolemia contributed to the pathogenesis of pregnancy-associated spontaneous coronary artery dissection (P-SCAD). In a systematic review of the literature, 16 women [median age 34 (31-36.5) years] with P-SCAD and angiographic follow-up were identified. The majority (69%) of P-SCAD cases occurred postpartum [median time after delivery: 13 (7-21) days]. In 10/16 (63%) patients medical treatment including betablocker and antiplatelet therapy was given leading to complete resolution of the dissection in 5 of them (31% of all patients) at follow-up, whereas in the other 5 patients the dissections were persisting or even progressive. Of the medically treated patients, 80% were free of symptoms suggestive for ischemia at follow-up. In 5/16 patients percutaneous coronary intervention (PCI) was performed as first-line therapy. Three patients underwent coronary artery bypass grafting, which was performed primarily in one patient, and secondarily in two patients with persisting dissections and ongoing ischemic symptoms after previous medical treatment or PCI without stenting, respectively. In conclusion, medical treatment including ASA, clopidogrel and betablocker therapy results in an excellent clinical and angiographic result in approximately one third of patients with P-SCAD. PMID- 16382386 TI - Stent-supported angioplasty versus endarterectomy for carotid artery stenosis: evidence from current randomized trials. AB - BACKGROUND: Carotid artery stenting (CAS) for carotid artery stenoses is evolving as an alternative to carotid endarterectomy (CEA). However, the value of CAS is still a matter of debate. Therefore, we performed a metaanalysis of the randomized controlled clinical trials (RCT) on this issue. METHODS: RCTs were identified through searching MEDLINE, textbooks and by personal communication. RESULTS: Six finished RCTs on this issue could be identified, including 1263 patients, 628 randomized to CAS and 635 to CEA. The 30-day death or stroke rate was 8.0% (50/628) in patients treated with CAS compared to 6.1% (39/635) in CEA patients (OR=1.36, 95% CI: 0.88-2.11; p=0.17; p for heterogeneity=0.009). The rate of cranial nerve palsy was 7.1% in the CEA compared to 0% in the CAS group (p<0.0001). The rate of myocardial infarctions was reduced from 3.1 to 1% (OR=0.32, 95% CI: 0.12- 0.81; p=0.02; p for heterogeneity=0.49). The death or stroke rate during follow-up was 12.1% in patients treated with CAS compared to 12.2% in CEA patients (OR=0.99, 95% CI: 0.70-1.42; p=0.98; p for heterogeneity=0.02). CONCLUSION: The available RCT data on CAS vs. CEA suggest that both methods seem to be equally effective concerning short- and medium-term results, while CAS is associated with lower minor complications than CEA. However, because of the significant heterogeneity between the study outcomes, the results of the large RCTs underway should be awaited before it can be advised to use CAS in a broader perspective. PMID- 16382387 TI - [Position paper to the taking of quality controls for resting, exercise, and long term-ECG]. PMID- 16382395 TI - [Differential diagnosis and therapy of pediatric disorders of the hip]. PMID- 16382397 TI - [Rational diagnostic and therapy of renal colic in the year 2005--what's new?]. AB - The acute flank pain is the most frequent urological emergency. Patients with renal colic are usually treated in emergency care units or by their family doctors and require immediate diagnosis and treatment. Up to 10 % of the population is estimated to suffer from kidney colic at least once in their lifetime. Besides, renal colic can occur during pregnancy and childhood, which require special attention when deciding therapy. Differential diagnosis of acute flank pain contains a series of diseases which belong not only in the urological field but need adequate directly therapy. Particularly, these principles should give useful advice, wherever patients are treated without urological department. PMID- 16382398 TI - [Soft tissue enhancement with injectable fillers for correction of age related folds and wrinkles]. AB - Injectable fillers for facial soft tissue enhancement have been developed and used for decades for the correction of age related folds and wrinkles. Many of the disadvantages of xenogenic and prior exogenous materials have been overcome with the advent of autologous and synthetic alternative materials. Autologous and synthetic injectable fillers herald a new era in the treatment of the aging face. Therefore this article will give an in-depth look at the implant choice, surgical approach, and possible complications and will provide a review of current injectable fillers for age related facial soft tissue augmentation. PMID- 16382399 TI - [Mechanisms influencing cellular physiology as a concept of treatment for wound healing disturbances]. AB - AIM: Recent knowledge about repair mechanisms in different types of tissue is the basic of actual therapeutic efforts. Center of several experimental and clinical approaches is the influencing of angiogenesis with an also distinct meaning concerning wound healing. Therefore, application of growth factors, gene transfer, and employment of genetically manipulated cells often aim at angiogenesis. Nevertheless, manipulation of angiogenesis also leads to secondary problems such as hyperpermeability followed by impairment of local wound milieu. Our study was done to identify mechanisms to protect from disturbances of endothelial barrier function. METHOD: In a first experimental investigation on cultured endothelial cells, the influence of plasma-transglutaminase (Factor XIII) to endothelial barrier function was studied. In a second step, the influence of Factor XIII on wound healing properties was investigated in patients with a chronic venous ulceration. RESULTS: Activated Factor XIII (FXIIIA*) led to a dose-dependent reduction of endothelial cell permeability of 30 % compared to control with a maximum effect using 1 to 5 U/mL. Clinical investigation revealed a nearly complete reduction of wound secretion. CONCLUSION: Experimental studies revealed that activated Factor XIII stabilizes endothelial barrier under basic conditions as well as under conditions of induced hyperpermeability. Clinical study revealed that Factor XIII also distinctly reduces wound secretion. Therefore, plasma-transglutaminase may offer a new therapeutic option to treat the local or generalized leakage-syndrome. PMID- 16382400 TI - [Resection of lung metastases--risk or chance?]. AB - INTRODUCTION: The resection of pulmonary metastases can extend life of patients, even offering the possibility of cure. A risk analysis of the surgical procedure under the current practice of the thoracic surgery has been performed in this study. MATERIALS AND METHODS: Using a prolective method, data from 199 patients with 300 lung resections for pulmonary metastases done in our university department (1/1994-01/2003) were evaluated. Risk factors and therapy results were investigated in an uni- and multivariate analysis (Fishers Exact Test, Chi-Square and logistic regression). RESULTS: Overall lethality was 1 % and the total morbidity rate was 19 %. The 232 atypical resections were associated with 14 % morbidity rate, while the anatomical and combined ones were associated with 38 % (p = 0.0001). 19 patients revealed a FEV (1) < 1.6 l linked with a morbidity of 42 %. In patients without FEV (1) restriction morbidity was 18 % (p = 0.0172). Both, risk factors and the anatomical resection showed in the multivariate analysis a statistically significant increased morbidity. Additional risk factors (ASA > 2, age over 65, sex or cardial risk factors) did not influence postoperative morbidity. DISCUSSION: According to our data, no patient with good lung function should be withhold from atypical resection of pulmonary metastases offering the possibility of cure with low morbidity. However the indication should be critically evaluated if anatomical resection is required. PMID- 16382401 TI - [Resection of combined or sequential lung and liver metastases of colorectal cancer: indication for everyone?]. AB - Successful sequential resection of isolated hepatic and pulmonary metastases of colorectal cancer (crc) has been reported, however long-term results of large series are lacking. Therefore, we retrospectively analysed data of patients in whom sequential hepatic and pulmonary resection for metastases was performed. PATIENTS AND METHOD: From the records of our hospital we identified 25 patients (19.5 % of all patients operated for hepatic or 33 % for lung metastases due to crc) with colorectal cancer who had pulmonary and hepatic resection for metastatic disease between 1991 and 2002. 11 of these had primary colonic cancer and 14 rectal cancer. None of the patients died perioperatively. Long-term results were correlated with the staging of the primary tumour, the number of metastases, disease free interval between primary tumour operation and occurrence of metastatic disease. RESULTS: Five-year survival rate was 33.5 % following the resection of the first metastasis. Three year survival after resection of the second metastasis was 39 %. The disease free interval was 20 months (mean). Long term results were clearly influenced by the disease free interval: < 1 year (n = 6) median 50 months after resection of the crc; > 1 year median 90 months (n = 19). Further on R0 resection was important for long-term survival: Median survival was 32.5 (+/- 4.1) months following resection of the second metastasis but only 9.9 months after R > 0 resection. CONCLUSION: These results confirm that sequential resection of hepatic and pulmonary metastases can be performed with curative intention provided a systemic spread of the disease is excluded. The surgeon's opinion of resectability should be obtained in patients with such metastases before the patient is scheduled for palliative conservative treatment. PMID- 16382402 TI - [Rectal prolapse in adults--causes, diagnostic, treatment]. AB - Despite progress in modern surgery, the choice of the surgical procedure of rectal prolapse is regarded with controversy. Selection criteria between the abdominal or perineal approach or between rectopexy and resection rectopexy are not yet proven. This article gives a review of the literature about rectal prolapse and an analysis of the outcome of posterior rectopexy and resection rectopexy--partly conventionally and partly laparoscopically--in 25 patients with rectal prolapse III degrees and IV degrees. All except for one patient were examined during a mean follow-up of 5.5 (3.1) years for the rectopexy group and 2.1 (0.7) years for the resection rectopexy group. Recurrence occurred in one patient in each group respectively. There was no significant difference concerning the continence function (p = 0.32) and constipation (p = 0.36) between both groups. No mesh-related complications such as infection, fistula or rectum stenosis were observed. According to the review of the literature and our data, we believe that the choice of the operative procedure for rectal prolapse should be based on individual criteria. Fit patients should be offered laparoscopic procedures such as resection rectopexy and rectopexy without colonic resection. PMID- 16382403 TI - [Total extraperitoneal endoscopic hernioplasty (TEP) in the treatment of incarcerated and irreponible inguinal and femoral hernias]. AB - To estimate the value of TEP in the treatment of incarcerated and irreponible inguinal and femoral hernias more exactly we prospectively collected and evaluated the data of our clinic for the period of Oct. 1999 until Dec. 2003. In this period we performed in total 1 671 hernia repairs including 79 patients suffering from an incarcerated (n = 33) or irreponible (n = 46) inguinal or femoral hernia. Using only the TEP-technique we treated mainly the irreponible hernias (46 patients). In the combination of LAP (laparoscopy) and TEP (27 patients) the laparoscopy provided the possibility to classify as well the incarcerated tissue as the result of the reposition. With this combined technique we treated the majority of the incarcerated hernias. To confirm the recovery of the incarcerated tissue laparoscopy can be of high value at the end of the combined LAP + TEP (2 patients). Thus TEP was performed in 92 % of the cases. In 2 cases we performed a conventional hernioplasty and one operation was finished conventionally after switching from endoscopic to conventional procedure. In 2 patients we performed a laparoscopically supported resection of the incarcerated tissue without patch implantation. 1 patient acquired TAPP. The use of different operative techniques and their combinations demonstrates as well the possibility as the necessity of a differentiated and case adapted proceeding in the treatment of incarcerated hernias. Lethality with 1.2 % and early postoperative morbidity with only 5.0 % were low. The hospitalisation period was 4.7 d on average. Our results are comparable to results of literature and show that TEP-technique and combined TEP + LAP-technique are possible and reasonable for the treatment of incarcerated and irreponible hernias. PMID- 16382404 TI - [How should patients with completely resected gastrointestinal stromal tumours (GIST) be followed up?]. AB - Gastrointestinal stromal tumours are topical because of their uncertain biological behaviour and the potential of treatment with imatinib. In the following study we have examined which pattern of follow-up is both appropriate for detecting recurrences and cost-effective. PATIENTS AND METHODS: Between July 1997 and February 2004 we treated 43 patients diagnosed with a GIST. Patients with high risk (HR), intermediate risk (IR), or overtly malignant (OM) tumours were followed-up regularly. In 2004 we screened all patients independent of their risk of malignant disease with an ultrasound scan and endoscopy followed by endosonography. Further diagnostic procedures were carried out if necessary. RESULTS: Overall, we diagnosed recurrences in five out of 33 patients at risk (two in patients with OM, one in a patient with HR, and 2 in patients with IR according to the NIH criteria). The time period between resection of the primary tumour and recurrence ranged from 4.5 to 33 months. One of the patients with a recurrence was seen before the imatinib era, the other four were treated with imatinib mesylate. CONCLUSION: In our experience, regular follow-up should be restricted to patients with OM, HR, and IR GIST. We suggest that patients are initially seen in six months intervals for two years and annually for another three years thereafter. PMID- 16382405 TI - [The arteries of the humerus and their relevance in fracture treatment]. AB - BACKGROUND: The purpose of the current work was to study the arterial blood supply of the humerus to evaluate impairments of the blood supply after humeral fractures and different methods of fracture treatment. METHODS: Arterial blood supply of the humerus was studied by corrosion injection technique in 6 cadaver preparations and is compared with the existing literature. RESULTS: Humeral head arteries originate from the circumflex humeral arteries. The major vessel enters the greater tubercle. Another group of vessels accesses the head from medially. The nutrient artery for the shaft enters into the medial third of the shaft. At the distal humerus there are multiple anastomoses with penetrating branches into both condyles and proximal to the radial, coronoid, and olecranon fossae. CONCLUSIONS: Fragments of a proximal humeral fracture usually have an independent blood supply. The medial vessels to the head gain high importance in fractures of the anatomical neck. The nutrient artery lies regularly medial to the humeral mid shaft and is at risk at open reduction of this region. The abundant vessels at the entire distal humerus ensure sufficient vascularization of the fragments after fracture. PMID- 16382406 TI - [Results of operative therapy of idiopathic clubfoot using a dorsomedial approach with soft tissue preparation "a la carte"]. AB - INTRODUCTION: Several concepts have been published for therapy of idiopathic clubfoot. Actually the Cincinnati approach is the "gold-standard" in operative treatment of idiopathic clubfoot. Using this approach delayed wound healing and overcorrection are wellknown complications. During the last 30 years a dorsomedial approach is used in operative treatment of idiopathic clubfoot in our clinic. Using this approach a dorsal, medial and/or lateral release can be performed. Postoperatively a plaster cast is used. The following years a rigid therapy with orthosis and support is requested. PATIENTS AND METHOD: Between June 1986 and December 2000 130 clubfoot operations with soft tissue release were performed. 119/130 (91 %) patients could be clinically and radiologically followed-up after a mean of 4.5 years (min: 2, max: 17) including 65.5 % male and 34.5 % female patients. Average age at time of operation was 7.6 months. The findings were classified according to Dimeglio. To evaluate the clinical results the score according to Laaveg and Ponseti was used. X-rays with load (a. p. and lateral view) were evaluated preoperatively and at follow-up for a. p. and lateral talocalcaneal angle, talometatarsal-I-angle, calcaneometatarsal-V-angle and the angle of the first ray. Additionally complications and recurrences were documented. RESULTS: 21.8 % of the feet were classified IV degrees , 39.5 % III degrees and 38.7 % II degrees according to Dimeglio. Recurrent clubfoot was found in 7.6 % of all cases. All angles showed a significant improve. At latest follow up 95.6 +/- 9.2 points according to the score of Laaveg and Ponseti were achieved. Flat top talus was found in 37 cases. CONCLUSION: The dorsomedial approach allows an excellent correction of clubfeet with a low rate of complications and recurrences in comparison to other studies. PMID- 16382407 TI - [Residual particle free surfaces after shot peening in modular hip arthroplasty are feasible]. AB - INTRODUCTION: Shot peening with steel balls is used for introduction of compressive stress in the production of morse taper junctions of modular hip endoprostheses. After this procedure, significant contamination with residual particles can be detected, which may lead to increased corrosion and third body wear. Additionally, the European norm EN 12010 requires surfaces free of residual particles. The aim of this study is to evaluate the effects of several cleaning procedures with regard to complying with EN 12010. MATERIAL AND METHOD: The surfaces of 12 Ti6Al7Nb rods with a diameter of 15 mm was shot peened with steel balls (GS VERA, Fa. Wurth, Bad Friedrichshall, Germany). A field emission scanning electron microscopy (SEM, LEO 1525) was used for the detection of residual particles on the surface of the rods with a backscattered electron detector. After SEM examination, one sample each was cleaned using shot peening with sugar or dry ice and 10 rods using a pH dependent cleaning procedure. After these cleaning procedures, the samples were again analysed with the SEM for residual particle contamination. RESULTS: The surface of all rods showed a considerable contamination with steel particles after shot-peening with steel balls. After the cleaning procedure with dry ice or sugar steel contamination was lower, however, only the pH dependent cleaning procedure was able to achieve surfaces free of residual particles. DISCUSSION: For smooth shot-peened surfaces in modular hip arthroplasty Euro Norm EN 12010 can be fulfilled using shot peening with steel balls followed by a pH dependent cleaning procedure. PMID- 16382408 TI - [The surgeon Burghard Breitner. poet or healer?]. AB - Burghard Breitner, born on June 10, 1884 in Mattsee (Salzburg state) went to school in Salzburg. From autumn 1901 he studied medicine in Graz and Vienna, where he graduated on 1 (st) of June 1908. In the time of his studies he worked on literature and poetry. But finally he decided to become a surgeon. In World War I Breitner was a prisoner of war (POW) of Russia. During this period he used his medical expertise to help wounded soldiers and the local people. For this honourable gesture he was popularly known as "The Angel of Siberia". From 1932 he was Professor of Surgery in Innsbruck. Mainly he worked on the therapy of goiter. Breitner restructured the department of surgery because of scientific development. In 1951 he was a presidential candidate in Austria in an election, which he did not win. In autumn 1952 he became the Rector magnificus of the University of Innsbruck. Breitner died on 28 (th) of March 1956 in Innsbruck. PMID- 16382409 TI - [Treatment of multiple renal artery aneurysms--a case report to demonstrate treatment options]. AB - Renal artery aneurysms require selective treatment, depending on size, localisation, and morphology as well as on singular or multiple locations. Spontaneous course without sequelae was reported in smaller series, however, there is an indication for treatment of aneurysms without calcification, women in childbearing age and in cases of unilateral kidney. Besides endovascular procedures there are different surgical techniques of in vivo or ex vivo repair. Treatment of aneurysms of side branches is more complicated and requires ex vivo treatment in selected cases. Ex vivo preparation of the kidney offers nearly identical surgical complications as the renal transplantation, limiting its indication. The clinical course and treatment options of a 53 year old female demonstrates that even nephrectomy might be the treatment of first choice in case of normal contralateral kidney. PMID- 16382410 TI - [The non-tropical pyomyositis--a rare disease in the moderate climate zone]. AB - Pyomyositis is in the moderate climate zone a rare and acute bacterial infection of the skeletal muscles without an obvious origin of infection. Initial symptoms contain muscle pain and localized swelling and tension. As different causes are taken into account first, the diagnosis of a pyomyositis is often delayed. We report on three cases of pyomyositis and discuss diagnosis and therapy against the background of the available and recent literature. PMID- 16382411 TI - [Legal aspects of deadly acute appendicitis]. AB - Appendicitis is diagnosed by synoptic evaluation of typical symptoms, laboratory tests and sonography. The only therapy is a prompt operation. The main reason for appendicitis mortality is the condition not detected or not detected in time. The case of a 50 year old male who died from a non-detected perityphlitic appendicitis with abscesses in the liver is presented. Only if appendicitis can be reliably ruled out an operation is not indicated. The not performed surgical intervention is often interpreted as a treatment error by the courts. In doubt, an operation should therefore be performed. PMID- 16382412 TI - Crohn's disease and a gastrointestinal stromal tumor in an 81-year-old man--a rare coincidence. AB - We describe an 81-year old man presenting with ileus and two rare, pathologically distinct entities: gastrointestinal stromal tumor (GIST) and Crohn's disease. Within Meckel's diverticulum a polypous tumor, 3 cm in diameter, with preserved lumen was found. In the area of the terminal ileum, coecum and colon ascendens inflamed bowel loops were fixed together with fibrous adhesions. Partial resection of the ileum with end-to-end anastomosis and right hemicolectomy with resection of the terminal ileum and end-to-end ileotransverse anastomosis were performed. Pathohistological and immunohistochemical examination revealed the polypous tumor as GIST. Changes in terminal ileum, coecum and colon ascendens were referred as Crohn's disease. Although adenocarcinoma is well known in chronic, long-standing inflammatory bowel disease, other primary intestinal tumors are rare in those patients. Furthermore, Crohn's disease can be a part of differential diagnosis of ileus, also in such an old man. PMID- 16382413 TI - [Prevalence of GBV-C/HGV (HGV) in HIV-infected patients and potential influence of co-infection on the course of the disease.]. AB - AIM OF THE STUDY: Assessment of prevalence of GBV-C/HGV infection in HIV-infected patients and evaluating of a possible influence of GBV-C/HGV on the course of HIV infection by assessment of immunological and virological markers of progression of HIV infection. MATERIALS AND METHODS: We have investigated serum samples of 273 HIV- infected patients from AIDS Center of the University Hospital Na Bulovce Prague during 2002-2003. Our target was to assess the presence of markers of GBV C/HGV infection by semiquantitative HGV RNA evaluation and anti-E2 antibodies by ELISA testing. 271 of serum samples were tested for HGV RNA and 269 samples were tested for anti-E2 antibodies. HIV viral load and CD4 count were tested concurrently. We used Spearmans test to rule out the dependency of CD4 count and HIV viral load on HGV infection. RESULTS: 89 (33,3 %) of patients were positive in HGV RNA test and 101 (38,5 %) of patients were positive in anti E2-ELISA. No statistically significant effect of GBV-C/HGV infection was observed on CD4 count and HIV viral load in our cohort of patients. CONCLUSIONS: The effect of GBV C/HGV infection on predictive laboratory markers of HIV infection was not confirmed in our study. Further investigations regarding this subject seem to be necessary. PMID- 16382414 TI - [Problems of cytomegalovirus diagnostics by real-time polymerase chain reaction.]. AB - AIM OF THE STUDY: The main goal was to explain the discrepancies between two PCR methods used for detection of cytomegalovirus (CMV) in peripheral blood samples of patients of Department of Internal Medicine-Hematooncology, University Hospital, Brno. MATERIALS AND METHODS: In past we used exon 4 of major immediate early (MIE) gene as the target for quantitative detection of the CMV in clinical samples, but sometimes this method failed to detect the viral load in samples that were positively tested using less sensitive qualitative method targeting another region (exon 2-4) of the same gene. From January 2004 to January 2005 we totally tested samples from 363 patients. 64 patients were at least once CMV positive using quantitative method, but 20 patients were repeatedly false negative.To find the cause of this discrepancy we performed partial sequence analysis of this region (nt positions 2719-2919, GenBank M21295) and glycoprotein B (gB) genotyping. We sequenced samples from 35 patients-15 giving true positive (both in qualitative and quantitative method) and 20 giving false negative (negative in quantitative but positive in qualitative method) results in several consecutive blood samples. RESULTS: The 15 true positive samples were 100% homological, whereas all 20 false negative samples showed high degree of variation from the laboratory strain AD169. These changes are not random and indicate that the two groups of patients were infected by different CMV genotypes. Moreover, sequence alignment showed similarity to laboratory strains Toledo and Towne. No preferential concordance was observed between clinical context, MIE exon 4 sequence and gB groups. CONCLUSIONS: Because of high sequence variability exon 4 of MIE gene can not be used for routine diagnostics. Genetic varibility among the pathogenic strains may seriously affect its proper diagnostics. PMID- 16382415 TI - [Analysis of occurence of Staphylococcus aureus strains and their resistance to antibiotics in-patients hospitalized at the National Institute of Tuberculosis and Respiratory Diseases in the years 2001-2004.]. AB - PURPOSE OF THE STUDY: The authors analysed the incidence of S. aureus strains and their resistance to antibiotics in in-patients in order to obtain information for the management of an antibiotic policy, as well as for an emprical treatment of staphylococcal infections. MATERIAL AND METHODS: The authors determined the sensitivity of S. aureus strains to antibiotics. Data on patients, biological material, bacterial strains and their sensitivity to antibiotics were processed by the programme WHONET 5.3. RESULTS: During the period of the study no substantial changes were seen in the proportion of S. aureus resistance to most investigated antibiotics. The chosen denominator-the number of complete hospital stays-helped to establish the list of wards at risk on the basis of patient colonization and the onset of infections. The authors investigated the incidence of S. aureus strains in the different wards. CONCLUSIONS: When processing an analysis of the incidence of bacterial resistance to antibiotics the use of an appropriate denominator represents the solution of the problem-because of the small set of investigated strains the proportion of resistance cannot be computed. PMID- 16382416 TI - [Viral hepatitis D imported to the Czech Republic by the citizen from the Moldavian Republic.]. AB - Viral hepatitis D (HDV) is a serious acute inflammatory liver damage, which frequently is transmitting to the chronic phase, cirrhosis and hepatocellular carcinoma. This disease can be found in the Czech Republic only rarely. The authors present the case of viral hepatitis D imported to the Czech Republic from Moldavia in 28-year old man. According to the examinations of the markers of HBV and HDV, the most probably it is a HDV superinfection in patient after acute HBV with HBsAg carriers. PMID- 16382417 TI - [Malaria-an unusual cause of fever in the infant.]. AB - Malaria is the fourth most frequent cause of death in African children. Connected with perinatal diseases as well as gastrointestinal and respiratory infections malaria has been still a serious health problem of that region. Occurrence of tropical malaria in infants reported in European countries is relatively rare. Not only from that reason, the assesment of diagnosis in children under one year of age seems to be obviously more difficult. The authors report the malaria in five-month-old infant from Cameroon who became ill during his stay in the Czech Republic. Non-specific symptoms, high level of parasitemia and impairment of blood coagulation were the main features of the emergent infection. On conclusion, the lack of suitable forms of childrens antimalarial drugs both for profylaxis and treatment is mentioned. PMID- 16382418 TI - Using pseudo-data to correct for publication bias in meta-analysis. AB - In many ways, adjustment for publication bias in meta-analysis parallels adjustment for ascertainment bias in genetic studies. We investigate a previously published simulation-based method for dealing with complex ascertainment bias and show that it can be modified for use in meta-analysis when publication bias is suspected. The method involves simulating sets of pseudo-data under the assumed model using guesses for the unknown parameters. The pseudo-data are subjected to the same selection criteria as are believed to have operated on the original data. A conditional likelihood is then used to estimate the adjusted values of the unknown parameters. This method is used to re-analyse a published meta analysis of the effect of the MTHFR gene on homocysteine levels. Simulation studies show that the pseudo-data method is unbiased; they give an indication of the number of pseudo-data values required and suggest that a two-stage adjustment produces less variable estimates. This method can be thought of as an example of the selection model approach to publication bias correction. As the selection mechanism must be assumed, it is important to investigate the sensitivity of any conclusions to this assumption. PMID- 16382420 TI - Estimating probit models with self-selected treatments. AB - Outcomes research often requires estimating the impact of a binary treatment on a binary outcome in a non-randomized setting, such as the effect of taking a drug on mortality. The data often come from self-selected samples, leading to a spurious correlation between the treatment and outcome when standard binary dependent variable techniques, like logit or probit, are used. Intuition suggests that a two-step procedure (analogous to two-stage least squares) might be sufficient to deal with this problem if variables are available that are correlated with the treatment choice but not the outcome. This paper demonstrates the limitations of such a two-step procedure. We show that such estimators will not generally be consistent. We conduct a Monte Carlo exercise to compare the performance of the two-step probit estimator, the two-stage least squares linear probability model estimator, and the multivariate probit. The results from this exercise argue in favour of using the multivariate probit rather than the two step or linear probability model estimators, especially when there is more than one treatment, when the average probability of the dependent variable is close to 0 or 1, or when the data generating process is not normal. We demonstrate how these different methods perform in an empirical example examining the effect of private and public insurance coverage on the mortality of HIV+ patients. PMID- 16382421 TI - On Bayesian analysis of misclassified data from a matched case-control study with a validation sub-study by Gordon J. Prescott and Paul H. Garthwaite. PMID- 16382422 TI - The Child Behavior Checklist as an indicator of posttraumatic stress disorder and dissociation in normative, psychiatric, and sexually abused children. AB - Expert ratings and confirmatory factor analyses were used to derive a posttraumatic stress disorder (PTSD), dissociation, and a combined PTSD/dissociation scale from the Child Behavior Checklist (CBCL). Validity was established by examining the relationship of these scales to features of sexual abuse thought to relate to severity and chronicity, as well as to self-report scales of PTSD and dissociation. In addition, this study examined differences between normative, psychiatric, and sexually abused children on the new scales. Both the sexual abuse and psychiatric sample differed significantly from the normative sample on all scales, but not from each other. Despite correlations of the dissociation and PTSD/dissociation combined scale with features of trauma and child self-report of PTSD and dissociation, the absence of differences between the clinical groups on the derived scales suggests that the scales measure generic, as opposed to trauma-related, distress. PMID- 16382423 TI - A randomized controlled trial of cognitive-behavior therapy for Cambodian refugees with treatment-resistant PTSD and panic attacks: a cross-over design. AB - We examined the therapeutic efficacy of a culturally adapted cognitive-behavior therapy for Cambodian refugees with treatment-resistant posttraumatic stress disorder (PTSD) and comordid panic attacks. We used a cross-over design, with 20 patients in the initial treatment (IT) condition and 20 in delayed treatment (DT). Repeated measures MANOVA, Group & times; Time ANOVAs, and planned contrasts indicated significantly greater improvement in the IT condition, with large effect sizes (Cohen's d) for all outcome measures: Anxiety Sensitivity Index (d = 3.78), Clinician-Administered PTSD Scale (d = 2.17), and Symptom Checklist 90-R subscales (d = 2.77). Likewise, the severity of (culturally related) neck-focused and orthostasis-cued panic attacks, including flashbacks associated with these subtypes, improved across treatment. PMID- 16382424 TI - Confirmatory factor analysis of posttraumatic stress symptoms in sexually harassed women. AB - Posttraumatic stress disorder (PTSD) factor analytic research to date has not provided a clear consensus on the structure of posttraumatic stress symptoms. Seven hypothesized factor structures were evaluated using confirmatory factor analysis of the Posttraumatic Stress Disorder Checklist, a paper-and-pencil measure of posttraumatic stress symptom severity, in a sample of 1,218 women who experienced a broad range of workplace sexual harassment. The model specifying correlated re-experiencing, effortful avoidance, emotional numbing, and hyperarousal factors provided the best fit to the data. Virtually no support was obtained for the Diagnostic and Statistical Manual of Mental Disorders, fourth edition (DSM-IV; American Psychiatric Association, 1994) three-factor model of re experiencing, avoidance, and hyperarousal factors. Different patterns of correlations with external variables were found for the avoidance and emotional numbing factors, providing further validation of the supported model. PMID- 16382425 TI - Use of sexual intercourse to reduce negative affect as a prospective mediator of sexual revictimization. AB - Increased risk of adult sexual assault (ASA) among women who experienced child sexual abuse (CSA) is well established. The strategies these women use to reduce negative affect secondary to CSA, such as sexual contact, may mediate the link between CSA and later ASA. Two waves of data from a racially diverse sample (i.e., 46% Black, 46.1% White, 7.9% other) of community women (N = 776) were analyzed using structural equation modeling. A history of CSA was associated with a twofold increase in the likelihood of experiencing ASA between the two measurement occasions (approximately 6 years). Psychological distress (i.e., depression, anxiety) and use of sex to reduce negative affect partially mediated the relation between CSA and prospective ASA. Implications for the treatment of CSA and prevention of sexual assault are discussed. PMID- 16382426 TI - Personality constellations in patients with a history of childhood sexual abuse. AB - Although childhood sexual abuse (CSA) appears to have an impact on personality, it does not affect all survivors the same way. The goal of this study was to identify common personality patterns in women with a history of CSA. A national sample of randomly selected psychologists and psychiatrists described 74 adult female patients with a history of CSA and a comparison group of 74 without CSA using the Shedler-Westen Assessment Procedure-200 (SWAP-200), a Q-sort procedure for assessing personality pathology. Q-factor analysis identified four personality constellations among abuse survivors: Internalizing Dysregulated, High Functioning Internalizing, Externalizing Dysregulated, and Dependent. The four groups differed on diagnostic, adaptive functioning, and developmental history variables, providing initial support for the validity of this classification. The data have potential methodological and treatment implications. PMID- 16382428 TI - Prolonged Exposure versus Eye Movement Desensitization and Reprocessing (EMDR) for PTSD rape victims. AB - This controlled study evaluated the relative efficacy of Prolonged Exposure (PE) and Eye Movement Desensitization and Reprocessing (EMDR) compared to a no treatment wait-list control (WAIT) in the treatment of PTSD in adult female rape victims (n = 74). Improvement in PTSD as assessed by blind independent assessors, depression, dissociation, and state anxiety was significantly greater in both the PE and EMDR group than the WAIT group (n = 20 completers per group). PE and EMDR did not differ significantly for change from baseline to either posttreatment or 6-month follow-up measurement for any quantitative scale. PMID- 16382429 TI - Exposure utilization and completion of cognitive behavioral therapy for PTSD in a "real world" clinical practice. AB - This study assessed rates of imaginal exposure therapy (ET) utilization and completion of cognitive behavioral therapy (CBT) for posttraumatic stress disorder (PTSD) in a clinical setting and examined variables associated with CBT completion. Using a clinical definition, the completion rate of CBT was markedly lower than rates reported in randomized trials. CBT completion was inversely related to severity of overall pretreatment measures of PTSD, avoidance, hyperarousal, depression, impaired social functioning, and borderline personality disorder. Regression yielded avoidance and depression as unique predictors of completion. Most dropouts occurred before starting imaginal ET, although initiating ET was associated with greater likelihood of completion. Results highlight methodological differences between research and practice notions of treatment completion and the need for further study of variables influencing CBT completion in practice settings. PMID- 16382430 TI - Factor structure of posttraumatic stress among Western New York undergraduates following the September 11th terrorist attack on the World Trade Center. AB - The structure of posttraumatic stress is of both theoretical and clinical interest. In the present study, seven models of posttraumatic stress were compared using confirmatory factor analysis. A sample of 528 Western New York undergraduate students was assessed 1 and 3 months after the September 11th, 2001 terrorist attacks. At the Month 1 assessment, the current three-factor Diagnostic and Statistical Manual of Mental Disorders (DSM-IV; American Psychiatric Association, 1994) model, which consists of Intrusions, Avoidance/Numbing, and Hyperarousal, did not provide a good fit to the data; however, a four-factor model consisting of factors labeled Intrusions, Avoidance, Dysphoria, and Hyperarousal did fit the data well and provided better fit than the three-factor model and other competing models. Importantly, Dysphoria spans symptoms from the traditional DSM Numbing and Hyperarousal clusters. The four-factor model continued to fit the data well at Month 3. These findings parallel the results of earlier studies which suggest that a four-factor model better reflects the nature of posttraumatic stress than do simpler models, including the DSM. The present work is consistent with a dimensional model of stress responses and calls for further longitudinal work in this area. PMID- 16382431 TI - Positive and negative changes following occupational death exposure. AB - Professionals who work in situations that expose them to death have long been of interest to traumatic stress research. However, the positive changes that these professionals may also experience have not been the subject of empirical scrutiny. This study examined occupational death exposure, death attitudes, subjective appraisals, intrusions, avoidance, social support, and positive and negative affect, and their associations with positive and negative psychological changes in funeral directors. Multivariate hierarchical regression analyses revealed that positive changes were significantly and independently predicted by an approach acceptance death attitude and social support; negative changes were significantly and independently predicted by fear of death, intrusions and avoidance, and occupational death exposure. The discussion focuses on how these findings extend the literature dealing with occupational death exposure, together with a consideration of limitations of the study that inform directions for future research. PMID- 16382432 TI - Who seeks treatment after a traumatic event and who does not? A review of findings on mental health service utilization. AB - This review aimed to identify factors associated with seeking treatment from mental health services after a traumatic event. Databases of literature were searched in a systematic manner and 24 relevant articles were found. Although many of the findings are inconsistent, the most important factors associated with treatment seeking appear to be a higher level of psychopathology, the type and level of the traumatic event, and sociodemographic characteristics, in particular female gender. Even though the evidence is insufficient to guide service development, suggestions for future research are made. The methodological quality of research should be improved to establish whether the inconsistency of findings reflects methodological artefacts or true differences between different samples and contexts. PMID- 16382433 TI - An Internet-based self-change program for traumatic event related fear, distress, and maladaptive coping. AB - This study compared the efficacy of an Internet-based, 8-week self-help program for traumatic event-related consequences (SHTC) (n = 13) to a wait-list (WL) condition (n = 14). The SHTC consisted of cognitive-behavioral modules that progressed from the least anxiety-provoking component (i.e., information) to the most anxiety-provoking (i.e., exposure). Participants were those who had experienced a traumatic event and had been experiencing subclinical levels of symptoms associated with the event. Participants mastered the material in each module before proceeding to the next module. Pre- and post-treatment assessments revealed that SHTC participants decreased avoidance behavior, frequency of intrusive symptoms, state anxiety, and depressive symptoms, and increased coping skills and coping self-efficacy significantly more than WL participants. SHTC participants demonstrated more clinically significant improvement than WL individuals. PMID- 16382434 TI - Testing the latent structure of posttraumatic stress disorder: a taxometric study of combat veterans. AB - Since the diagnosis of posttraumatic stress disorder (PTSD) first appeared in the psychiatric nomenclature in 1980, considerable debate has revolved around the nature of the condition. Specifically, is PTSD best conceptualized as one end of a continuum of human response to traumatic stress or does it represent a discontinuous latent category? Two taxometric procedures were used to investigate this issue in a random community sample of 692 Australian combat veterans, using structured interview and self-report instruments to assess PTSD symptomatology. Findings favored a dimensional model of PTSD, consistent with previous taxometric work on treatment-seeking samples (A. Ruscio, Ruscio, & Keane, 2002). Implications are drawn for the conceptualization, etiology, and assessment of PTSD. PMID- 16382435 TI - Multisample cross-validation of a model of childhood posttraumatic stress disorder symptomatology. AB - This study is the latest advancement of our research aimed at best characterizing children's posttraumatic stress reactions. In a previous study, we compared existing nosologic and empirical models of PTSD dimensionality and determined the superior model was a hierarchical one with three symptom clusters (Intrusion/Active Avoidance, Numbing/Passive Avoidance, and Arousal; Anthony, Lonigan, & Hecht, 1999). In this study, we cross-validate this model in two populations. Participants were 396 fifth graders who were exposed to either Hurricane Andrew or Hurricane Hugo. Multisample confirmatory factor analysis demonstrated the model's factorial invariance across populations who experienced traumatic events that differed in severity. These results show the model's robustness to characterize children's posttraumatic stress reactions. Implications for diagnosis, classification criteria, and an empirically supported theory of PTSD are discussed. PMID- 16382436 TI - Posttraumatic stress disorder symptoms among low-income, African American women with a history of intimate partner violence and suicidal behaviors: self-esteem, social support, and religious coping. AB - There is a dearth of research on risk/protective factors for posttraumatic stress disorder (PTSD) among low-income African American women with a history of intimate partner violence (IPV), presenting for suicidal behavior or routine medical care in a large, urban hospital. We examined self-esteem, social support, and religious coping as mediators between experiences of child maltreatment (CM) and IPV and symptoms of PTSD in a sample (N = 134) of low-income African American women. Instruments used included the Index of Spouse Abuse, the Childhood Trauma Questionnaire, the Taylor Self-Esteem Inventory, the Multidimensional Profile of Social Support, the Brief Religious Coping Activities Scale, and the Davidson Trauma Scale. Both CM and IPV related positively to PTSD symptoms. Risk and resilience individual difference factors accounted for 18% of the variance in PTSD symptoms over and above IPV and CM, with self-esteem and negative religious coping making unique contributions. Both variables mediated the abuse-PTSD symptom link. In addition, we tested an alternate model in which PTSD symptoms mediated the relationship between abuse and both self-esteem and negative religious coping. PMID- 16382437 TI - Assessing violence exposure and trauma symptoms in young children: a critical review of measures. AB - The acknowledgment of the existence of age-specific posttraumatic stress symptoms in infants, toddlers, and preschoolers points to the urgent need for standardized assessment tools for violence exposure and trauma symptoms in young children. The authors review the assessment measures currently available for the evaluation of potentially traumatic events (PTE) and posttraumatic stress disorder (PTSD) symptoms in children less than 6 years old. Each measure is described and its strengths and weaknesses discussed in a developmental context, while also considering the specific difficulties inherent to the assessment of young children. Recommendations for further test development are given. PMID- 16382438 TI - Psychometric characteristics of Spanish and English versions of the Civilian Mississippi scale. AB - Following the 1994 Northridge earthquake, the psychometric properties of the Civilian Mississippi scale (CMS) were examined in three ethnic language subgroups (non-Latino-English, Latino-English, and Latino-Spanish). The psychometric patterns corroborated the existence of two subscales: 25 items with regular wording and 10 items with reversed wording. Psychometric properties were examined in relation to demographics, earthquake exposure, trauma, and psychological distress as measured by the Brief Symptom Inventory. The results suggest that items with reversed wording generated significantly higher mean posttraumatic stress disorder (PTSD) scores and lower reliability. Latinos and Spanish-speaking respondents had particular difficulty understanding items with reversed wording, which correlates with significantly lower or no correlation between the CMS-PTSD and psychological distress. PMID- 16382439 TI - Posttraumatic stress symptoms, coping, and physical health status among university students seeking health care. AB - This study examined posttraumatic stress disorder (PTSD) symptoms, coping, and physical health status in students reporting a trauma history (N = 138) using structural equation modeling. Participants completed questionnaires assessing PTSD symptoms, coping specific to health-related and trauma-related stressors and physical health. After accounting for coping with health-specific problems, trauma-specific avoidance coping was uniquely associated with poorer health status. Posttraumatic stress disorder symptoms were associated with poorer physical health status, controlling for age, health behaviors, and other psychopathology. In addition, the effect of PTSD symptoms on poorer health status was mediated by health- and trauma-specific avoidance coping. Results suggest that university health centers should screen for PTSD and consider psychoeducational programs and coping skills interventions for survivors of trauma. PMID- 16382440 TI - Problem gambling in Australian PTSD treatment-seeking veterans. AB - This study explored gambling among Australian veterans entering posttraumatic stress treatment programs (n = 153). Twenty-eight percent reached the South Oaks Gambling Screen (SOGS) criteria for probable problem gambling, as did 17% on the DSM-IV gambling scale (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition; American Psychiatric Association, 1994). Almost all problem gamblers reported gambling to escape problems in other areas of their lives. The strongest independent predictor of problem gambling was gambling weekly or more often on electronic gaming machines. There was no significant relationship between problem gambling, posttraumatic stress disorder (PTSD), anxiety, depression, or alcohol use. The study identified an entrenched gambling culture among PTSD treatment-seeking veterans, finding these veterans indulge in many different forms of gambling and that these forms are mediated by situational factors that provide both casual and formal gambling opportunities. PMID- 16382441 TI - Chronic PTSD patients' functioning before and after the September 11 attacks. AB - This study examined how treatment-seeking veterans with preexisting posttraumatic stress disorder (PTSD) (n = 178) were affected by vicarious exposure to the September 11 terrorist attacks. Participants were surveyed 0 to 5 months prior to 9/11 and resurveyed 6 months after the attacks. Half the patients reported that thoughts and feelings about 9/11 impaired their functioning some (37%) or most or all of the time (13%). However, there was little evidence that vicarious exposure to 9/11 altered the course of these patients' functioning. Mean symptom, substance use, and role functioning outcomes were unchanged from pre-9/11 levels. Time spent following media coverage of 9/11 events was weakly associated (r = .17 to .18, p < .05) with only two of eight functioning outcomes. PMID- 16382442 TI - Na+-K+-ATPase is not involved in the warming-up phenomenon in generalized myotonia. AB - The initial temporary weakness that occurs in autosomal-recessive generalized myotonia diminishes with repetitive contractions. Physiological understanding of this phenomenon is incomplete. The underlying hypothesis of our study was that the "warming-up" phenomenon relates to the exercise-related activation of Na(+) K(+)-ATPase. Three patients performed isometric exercise of the brachioradialis muscle on two separate days. Randomly, on one of these days the contraction was preceded by a 30-min infusion of the Na(+)-K(+)-ATPase inhibitor ouabain into the brachial artery of the exercising arm (0.4 mug.min(-1).dl(-1)). Force was measured simultaneously with electrical muscle activity using high-density surface electromyography (HD-sEMG). A transient rapid decline in force occurred after initiation of exercise, accompanied by electrophysiological changes indicating sarcolemmal conduction block. Ouabain infusion did not affect the recovery from transient paresis or the accompanying electromyographic changes, indicating that the warming-up phenomenon in generalized myotonia is not mediated by Na(+)-K(+)-ATPase. PMID- 16382443 TI - Single-fiber electromyography in limb and facial muscles in muscle-specific kinase antibody and acetylcholine receptor antibody myasthenia gravis. AB - We examined the findings from single-fiber electromyography in extensor digitorum communis (EDC) and orbicularis oculi (OOc) in 13 myasthenia gravis (MG) patients with muscle-specific kinase antibodies (MuSK-MG) and 12 MG patients with acetylcholine receptor antibodies (AChR-MG) with similar clinical scores. More than 70% of AChR-MG patients had abnormal jitter in both EDC and OOc, but the majority of MuSK-MG patients had normal jitter in EDC despite abnormal jitter in OOc. These findings demonstrate clear differences between the neurophysiology of MuSK-MG and AChR-MG. PMID- 16382444 TI - Comparison of three algorithms for multi-motor unit detection and waveform marking. AB - Quantitative EMG (QEMG) techniques include automated motor unit action potential (MUAP) detection and marking of clinically useful waveform metrics. Different computer algorithms are available on modern EMG machines to perform these operations rapidly. However, the efficiency and accuracy of available algorithms are rarely directly compared. We have assessed three commercially available algorithms using both synthesized and biologic interference patterns and found differences among algorithms, some of which are clinically significant. Our results point out the importance of assessing for duplicate MUAPs (same waveform detected as two separate waveforms) and accuracy of markings used to determine MUAP metrics. PMID- 16382446 TI - H2AX is not affected in mantle cell lymphoma with 11q23 deletion. PMID- 16382445 TI - Regional activation of chromosomal arm 7q with and without gene amplification in taxane-selected human ovarian cancer cell lines. AB - Taxanes are important drugs in the treatment of ovarian and other cancers, but their efficacy is limited by intrinsic and acquired drug resistance. Expression of the multidrug transporter P-glycoprotein, encoded by the MDR1 (ABCB1) gene, is one of the causes of clinical drug resistance to taxanes. To study the mechanisms of MDR1 activation related to taxanes, we established 11 multidrug-resistant variants from six ovarian cancer cell lines by continuous exposure to either paclitaxel or docetaxel. We profiled gene expression and gene copy number alterations in these cell lines using cDNA microarrays and identified a cluster of genes coactivated with MDR1 in 7q21.11-13. Regional activation was evident in nine resistant variants displaying a coexpression pattern of up to 22 genes over an 8-Mb area, including SRI, MGC4175, CLDN12, CROT, and CDK6. In six of these variants, regional activation was driven by gene copy number alterations, with low-level gains or high-level amplifications spanning the involved region. However, three variants displayed regional increases in gene expression even without concomitant gene copy number changes. These results suggest that regional gene activation may be a fundamental mechanism for acquired drug resistance, with or without changes in gene dosage. In addition to numerical and structural chromosomal changes driven by genome instability in cancer cells, other mechanisms might be involved in MDR1 regional activation, such as chromatin remodeling and DNA or histone modifications of the 7q21 region. PMID- 16382447 TI - Shwachman syndrome as mutator phenotype responsible for myeloid dysplasia/neoplasia through karyotype instability and chromosomes 7 and 20 anomalies. AB - An investigation of 14 patients with Shwachman syndrome (SS), using standard and molecular cytogenetic methods and molecular genetic techniques, showed that (1) the i(7)(q10) is not, or not always, an isochromosome but may arise from a more complex mechanism, retaining part of the short arm; (2) the i(7)(q10) has no preferential parental origin; (3) clonal chromosome changes, such as chromosome 7 anomalies and del(20)(q11), may be present in the bone marrow (BM) for a long time without progressing to myelodysplastic syndrome (MDS)/acute myeloid leukemia (AML); (4) the del(20)(q11) involves the minimal region of deletion typical of MDS/AML; (5) the rate of chromosome breaks is not significantly higher than in controls, from which it is concluded that SS should not be considered a breakage syndrome; (6) a specific kind of karyotype instability is present in SS, with chromosome changes possibly found in single cells or small clones, often affecting chromosomes 7 and 20, in the BM. Hence, we have confirmed our previous hypothesis that the SS mutation itself implies a mutator effect that is responsible for MDS/AML through these specific chromosome anomalies. This conclusion supports the practice of including cytogenetic monitoring in the follow-up of SS patients. PMID- 16382448 TI - Identification of a new cancer/testis gene family, CT47, among expressed multicopy genes on the human X chromosome. AB - Cancer/testis (CT) genes are normally expressed in germ cells only, yet are reactivated and expressed in some tumors. Of the approximately 40 CT genes or gene families identified to date, 20 are on the X chromosome and are present as multigene families, many with highly conserved members. This indicates that novel CT gene families may be identified by detecting duplicated expressed genes on chromosome X. By searching for transcript clusters that map to multiple locations on the chromosome, followed by in silico analysis of their gene expression profiles, we identified five novel gene families with testis-specific expression and >98% sequence identity among family members. The expression of these genes in normal tissues and various tumor cell lines and specimens was evaluated by qualitative and quantitative RT-PCR, and a novel CT gene family with at least 13 copies was identified on Xq24, designated as CT47. mRNA expression of CT47 was found mainly in the testes, with weak expression in the placenta. Brain tissue was the only positive somatic tissue tested, with an estimated CT47 transcript level 0.09% of that found in testis. Among the tumor specimens tested, CT47 expression was found in approximately 15% of lung cancer and esophageal cancer specimens, but not in colorectal cancer or breast cancer. The putative CT47 protein consists of 288 amino acid residues, with a C-terminus rich in alanine and glutamic acid. The only species other than human in which a gene homologous to CT47 has been detected is the chimpanzee, with the predicted protein showing approximately 80% identity in its carboxy terminal region. PMID- 16382449 TI - Multicolor fluorescence in situ hybridization (SKY) in mycosis fungoides and Sezary syndrome: search for recurrent chromosome abnormalities. AB - Cutaneous T-cell lymphoma (CTCL) is a clonally derived lymphoproliferative disorder that preferentially involves the skin. The two major clinical expressions of CTCL, mycosis fungoides (MF) and Sezary syndrome (SS), have poorly understood pathogenesis. Chromosome abnormalities, mostly complex karyotypes, are seen in about 50% of patients with MF/SS, and there have only been a few instances of recurrent rearrangements. We analyzed 19 blood samples from patients with MF/SS with cytogenetics and multicolor FISH (SKY) to better describe the complex karyotypes and search for recurrent abnormalities or breakpoints. Comparison of phytohemagglutinin (PHA)-stimulated cultures versus a combination of interleukin 2 plus interleukin 7 showed similar efficiency in detecting abnormal clones; however, the PHA cultures yielded more analyzable metaphases. Nine of 19 patients (47%) had an abnormal karyotype. The most frequent abnormalities, in 7 of 9 cases, involved chromosome 10; followed by chromosome 6, in 6 of 9 cases; chromosomes 3, 7, 9, 17, and 19, in 5 of 9 cases; chromosomes 1 and 12, in 4 of 9 cases; and chromosomes 8, 11, and 13, in 3 of 9 cases. Most abnormalities were structural. Recurrent rearrangements included deleted chromosomes 6 and 13, in three cases each, and recurrent breakpoints at 1p32-36, 6q22-25, 17p11.2-13, 10q23-26, and 19p13.3, occurring in three or more cases. One patient had a pseudodicentric translocation between the short arms of chromosomes 8 and 17, confirmed by dual-color FISH and interpreted as psu dic(17;8)(p11.2;p11.2). Two patients with SS reported in the literature seem to have a similar translocation. If confirmed, a psu dic(17;8) could be the first recurring translocation detected in at least three patients with MF/SS. PMID- 16382450 TI - Balloon dilatation vs. balloon dilatation plus bile duct endoprostheses for treatment of anastomotic biliary strictures after liver transplantation. AB - Biliary strictures after liver transplantation are a therapeutic challenge for endoscopy. Anastomotic strictures occur in 10% of patients after liver transplantation, leading untreated to mortality and ultimately to graft failure. Despite of successful reports, to date, there is no defined endoscopic therapy regimen for these cases. Therefore the aim of this study was to determine the most suitable concept for endoscopic treatment of post-liver transplant anastomotic strictures (PTAS). A total of 72 patients post-liver transplantation, who received endoscopic retrograde cholangiography (ERC) as a consequence of suspected biliary complications were retrospectively screened for the presence of PTAS. In all patients graft rejection or bile duct ischemia were excluded prior to ERC by liver biopsy or Doppler ultrasound respectively. We compared either balloon dilatation (BD) alone or dilatation plus placement of an increasing number of bile duct endoprostheses (BD + endoprostheses) in a retrospective analysis. A total of 25 of 75 patients showed PTAS. Overall, endoscopic therapy was successful in 22 of 25 patients (88%). BD was initially successful in 89% but showed recurrence in 62%. BD + endoprostheses was initially successful in 87%, and recurrence was observed only in 31%. All recurrences were successfully retreated by BD + endoprostheses. During 22 of 109 (20%) treatment sessions stone extraction was necessary. Complication rate was low with bacterial cholangitis in 8 of 109 (7.3%) sessions, mild pancreatitis in 10 of 109 (9%) sessions and minor bleeding in 2 of 25 (8%) sphincterotomies. Median follow-up after conclusion of endoscopic therapy is 6 months (range 1-43). In conclusion, our data confirm that endoscopic therapy of PTAS is highly effective and safe. As primarily successful BD shows a high rate of recurrence, we recommend a combination of BD followed by an increasing number and diameter of endoprostheses. Therapy sessions are effective at short intervals of every 2-3 months. PMID- 16382451 TI - Viral and fungal infections after liver transplantation--part II. AB - Viral and fungal infections in liver transplant recipients are important to recognize and treat early because of their association with substantial morbidity and mortality. Some viruses, such as cytomegalovirus and human herpesvirus 6, have immunomodulatory properties and can facilitate other infections, including fungal infections. Cytomegalovirus has long been recognized as an important virus in transplantation, but in the past decade other viruses have also received attention in the medical literature because of their association with particular clinical syndromes. Although human herpesvirus 6 has been associated with fever, rash, and encephalitis, a direct cause-and-effect relationship is still lacking. Human herpesvirus 8 has been found to be the cause of Kaposi sarcoma. Molecular techniques (e.g., pp65 antigenemia and polymerase chain reaction) that have been introduced for routine diagnosis of viruses have facilitated the diagnosis of asymptomatic viral infections and the institution of preemptive therapy. Nonetheless, the diagnosis of invasive fungal infections in liver transplant recipients is often delayed and thus associated with high mortality. Despite the use of new antifungal agents in clinical practice and the reduced incidence of fungal infections because of antifungal prophylaxis regimens, mortality has not decreased. Future patient outcomes may improve with early identification of patients who have risk factors for invasive fungal infections and with the development of new molecular diagnostic techniques for early detection. PMID- 16382452 TI - Mortality risk, behavior, and pediatric liver allocation. PMID- 16382453 TI - Bigger may not be better when it comes to hepatocytes. PMID- 16382454 TI - The challenge of progressive hepatitis C following liver transplantation. PMID- 16382455 TI - Measurement of health-related quality of life in patients with chronic liver disease. PMID- 16382456 TI - Validity of the Spanish version of the Chronic Liver Disease Questionnaire (CLDQ) as a standard outcome for quality of life assessment. AB - The Chronic Liver Disease Questionnaire (CLDQ) measures the impact on quality of life of chronic liver diseases, regardless of underlying etiology. The aim of this study was to develop a Spanish version of the CLDQ, and to assess its acceptability, reliability, validity, and sensitivity to change. The forward and back-translation method by bilingual translators, with expert panel and pilot testing on patients, was used for the adaptation. The final version was self administered, together with the Short Form-36 Health Survey (SF-36), on 149 consecutive patients with chronic liver disease. Child-Turcotte-Pugh scores were evaluated by a physician. To assess reproducibility and responsiveness the CLDQ was readministered to a subsample of stable patients and to those who had received a liver transplant. Validity was evaluated via exploratory factor analysis, the CLDQ pattern across severity groups, and correlation coefficients with "itching" and SF-36 scores. Cronbach's alpha and Intraclass Correlation Coefficient for CLDQ global score were 0.93 and 0.90, respectively, demonstrating good reliability. Validity was supported by correlations of the CLDQ with SF-36 and "itching," and CLDQ severity gradient (global score means were 5.5, 5.2, 5.0, and 4.5 in patients with no cirrhosis, cirrhosis Child-Turcotte-Pugh A, B, and C, respectively; P = 0.012). Responsiveness was shown by a high CLDQ improvement in patients who had received liver transplant (mean change = -1.4; P < 0.001). In conclusion, the Spanish CLDQ is reliable, valid, responsive, and equivalent to the original. These findings support its use as a standard outcome for patients with chronic liver diseases within the whole severity range, from "no cirrhosis" to transplant recipients, both in Spanish and international studies. PMID- 16382457 TI - Plasma total homocysteine and cardiovascular risk in patients submitted to liver transplantation. AB - Patients submitted to orthotopic liver transplantation (OLT) show an increased rate of cardiovascular events. OLT subjects have high homocysteine (Hcy) levels, but no data are available on the association of Hcy with cardiovascular events. In a cross-sectional analysis, 230 subjects were studied at least 6 months after OLT (159 on cyclosporine, 71 on tacrolimus). Routine laboratory data and total Hcy were recorded, as well as the history of diabetes, hypertension, dyslipidemia, and overweight. Cardiovascular events occurring in a follow-up of 2 36 months were registered. OLT subjects had higher-than-normal Hcy (median 16.7 micromol/L, range 6.1-171.8) without difference between the 2 immunosuppressive agents. The prevalence of Hcy >15 micromol/L was also similar, and significantly correlated with creatinine levels. A total of 28 arterial events occurred in 25 patients during follow-up (11 in coronary arteries, 10 in peripheral arteries, and 7 in splanchnic arteries). Deep vein thromboses occurred in 2 patients, and splanchnic vein thromboses in 4 patients. Cardiovascular events were frequently associated to high Hcy and hypertension. Cox regression analysis showed that high Hcy was significantly associated with arterial events. The risk of any arterial event, coronary artery or peripheral artery event increased by nearly 10% for any increase in Hcy of 5 micromol/L. In conclusion, high Hcy may be involved in the pathogenesis of cardiovascular events in OLT patients. The usefulness of Hcy lowering therapy remains to be verified. PMID- 16382458 TI - Efficacy and safety of low-dose valganciclovir in the prevention of cytomegalovirus disease in adult liver transplant recipients. AB - The efficacy and safety of valganciclovir (VGCV) for cytomegalovirus (CMV) prophylaxis in liver transplant recipients has not been established. We retrospectively compared the efficacy and safety of low-dose oral VGCV (450 mg once daily for 90 days) and standard oral ganciclovir (1 g three times a day for 90 days, GCV) in preventing CMV disease in 109 adult liver transplant recipients who survived at least 1 month between January 2001 and April 2003 (49 GCV and 60 VGCV). The incidence of CMV disease at 1 year post-transplant was similar among patients treated with VGCV and GCV (3% and 4%, respectively). Three of the four CMV disease cases occurred in high-risk recipients with CMV serotype of donor+/recipient- (D+/R-) and all cases presented after completion of CMV prophylaxis, ranging 114-152 days post-transplant. Severe neutropenia was rare, and thrombocytopenia and anemia occurred at similar frequencies with both prophylaxis regimens. In conclusion, a 90-day regimen of low-dose oral VGCV has a similar efficacy and safety profile to high-dose oral GCV in adult liver transplant recipients. D+/R- liver transplant recipients remain at risk of developing CMV disease after completion of antiviral prophylaxis. Additional prospective studies with close monitoring for CMV viremia and drug resistance are needed to further establish the optimal dose and duration of VGCV in liver transplant recipients. PMID- 16382459 TI - Coccidioidomycosis in liver transplantation. AB - Coccidioidomycosis is an endemic fungal infection in the southwestern United States. It causes morbidity and mortality among solid organ transplant recipients who reside in or visit the endemic area or who receive organs from donors infected with the fungus. This paper reviews current literature addressing these infections in liver transplantation programs, including risk factors, clinical manifestations in persons with cirrhosis or who have had a liver transplantation, prophylaxis, treatment, and outcomes. PMID- 16382460 TI - National and regional analysis of exceptions to the Pediatric End-Stage Liver Disease scoring system (2003-2004). AB - Since February 2002, the Pediatric End Stage Liver Disease (PELD) scoring system has been utilized as a means of prioritizing children for liver transplantation. The United Network for Organ Sharing database was queried to assess utilization of PELD in 2003 and 2004; 682 liver transplants were performed in pediatric recipients where the PELD score was potentially the primary determinant of liver allocation. In the majority of circumstances (53%) the actual calculated PELD score was not utilized to determine liver allocation. An exception to the PELD score was utilized in 24% of cases. An additional 29% of the children were listed as urgent (status 1) without having acute liver failure. There was considerable regional variability in the inability to utilize actual PELD scores for liver allocation to children. PELD utilization was higher in regions of the country where pediatric donor organs were more available, presumably because children have some priority for organs from pediatric donors. There were 87 deaths in children awaiting liver transplantation. The mean PELD score in children without acute liver failure or metabolic liver disease (n = 33) near the time of death was 24.2, which has a purported 3-month risk of mortality of less than 10%. In our opinion the assigned 3-month risk of mortality associated with PELD scores is understated. Three-month mortality risk is used to inter-convert the adult and pediatric scoring systems. Therefore exceptions to the scoring system are required when children compete with adults for donor organs. In conclusion, urgent reassessment of the PELD scoring system is needed to avoid morbidity and mortality in children. PMID- 16382461 TI - Effect of low central venous pressure and phlebotomy on blood product transfusion requirements during liver transplantations. AB - Correction of coagulation defects with plasma transfusion did not decrease the need for intraoperative red blood cells (RBC) transfusions during liver transplantations. On the contrary, it led to a hypervolemic state that resulted in an increase of shed blood. As well, plasma transfusion has been associated with a decreased one-year survival rate. The aim of the present prospective survey was to evaluate whether anesthesiologists could reduce intraoperative RBC transfusions during liver transplantations by changing their anesthesia practice, more specifically by maintaining a low central venous pressure (CVP), through restriction of volume replacement, elimination of all plasma transfusion and by using intraoperative phlebotomy during the transplantation. One hundred consecutive liver transplantations were prospectively studied during a two-year period and were compared to a retrospective series (1998-2002). A low CVP was maintained in all patients prior the anhepatic phase. Coagulation disorders were not corrected preoperatively, intraoperatively, or post-operatively unless uncontrollable bleeding. Phlebotomy and Cell Saver (CS) were used following pre established criteria. Independent variables were analyzed in a univariate and multivariate fashion. The mean number of intraoperative RBC units transfused was 0.4 +/- 0.8. No plasma, platelets, albumin, or cryoprecipitate were transfused. Seventy-nine percent of the patients received no blood products during their liver transplantation. The average final hemoglobin value was 85.9 +/- 17.8 g/L. In 57 patients (58.2%), intraoperative phlebotomy and CS were used either together or separately. The one-year year survival rate was 89.1%. Logistic regression showed that avoidance of plasma transfusion, starting hemoglobin value and phlebotomy were significantly linked to liver transplantation without RBC transfusion. In conclusion, the avoidance of plasma transfusion and maintenance of a low CVP prior to the anhepatic phase were associated with a decrease in RBC transfusions during liver transplantations. Previous reports indicating that it is neither useful nor necessary to correct coagulation defects with plasma transfusion prior to liver transplantation are further corroborated by this prospective survey. We believe that this work also supports the practice of lowering CVP with phlebotomy in order to reduce blood loss, during liver dissection, without any deleterious effect. PMID- 16382462 TI - Hepatic artery reconstruction with double-needle microsuture in living-donor liver transplantation. AB - In living-donor liver transplantation (LDLT), reconstruction of the hepatic artery is challenging because the recipient artery is located deep in the abdominal cavity and the operating field is limited. Also, the hepatic artery of the graft is short and the recipient artery is occasionally damaged. To overcome these difficulties, we developed a double-needle microsuture technique for artery reconstruction. A total of 161 adult patients received 163 LDLTs using this new technique. The first suture was placed at the most difficult point in the artery to be visualized through the microscope. Each stitch was placed from the inner side of the arterial wall to the outer side. The posterior stitch was tied pulling toward the back. The subsequent sutures were advanced anteriorly on either side adjacent to the previous suture. Hepatic artery thrombosis occurred in 4 patients (2.5%), only 2 (1.2%) of which were associated with arterial reconstruction. Intimal dissection developed in the recipient artery in 2 patients (1.2%). Three (50%) of these 6 complications occurred more than 10 days after LDLT. In conclusion, this suturing technique allows for safe intimal adaptation even when the arterial tunica intima is separated from the tunica media, because all stitches are carried from inside of the vessel to the outside, contributing to more satisfactory results. PMID- 16382463 TI - Protective antibody levels and dose requirements for IV 5% Nabi Hepatitis B immune globulin combined with lamivudine in liver transplantation for hepatitis B induced end stage liver disease. AB - Lamivudine combined with Hepatitis B immune globulin (HBIg) prevents post liver transplant (LT) HBV recurrence. The study was designed to assess the impact of lamivudine on hepatitis B antibody (anti-HBs) and dosage requirements of intravenous 5% HBIg (Nabi-HB) in the first 36 weeks post LT. Adults undergoing LT for chronic HBV received lamivudine prior to or at LT, and IV HBIg 20,000 IU on day of LT, 10,000 on days 1-7, weeks 4 and 8, and 5,000 every 4 weeks thereafter. Replicative status based on serum HBV DNA (> 5 pg/mL = replicator (R) or < or = 5 pg/mL = nonreplicator (N) was determined at initiation of lamivudine (R or N) and within 2 weeks of LT (r or n), resulting in 3 groups: Nn, Rn, and Rr. Between December 1999 and May 2001, 30 patients (10 Nn, 13 Rn, 6 Rr, and 1 unknown), mean age of 52 years underwent LT. HBsAg neutralization was achieved with anti-HBs > 300 IU/L during week 1 and > 200 IU/L during weeks 2-12. All but one patient were HBsAg-negative on last follow-up. Pre-LT suppression of HBV replication resulted in similar dose requirements and pK in the Rn and Nn groups within 1 week after LT. Comparatively, the Rr group had greater HBIg requirements during weeks 1-12 due to greater anti-HBs clearance and shortened t(1/2) during the entire 36-week follow-up. In conclusion, this study provides a rationale for the use of lower HBIg doses in HBV patients with suppressed replication undergoing LT. PMID- 16382464 TI - Cyclosporine suppresses hepatitis C virus in vitro and increases the chance of a sustained virological response after liver transplantation. AB - Cyclosporine is an immunosuppressive agent widely used in the management of liver transplant recipients. Cyclosporine has been shown to have antiviral activities against HIV, herpes simplex, and vaccinia viruses. The aim of this study was to determine the effect of Cyclosporine in viral clearance in the liver transplant recipients during therapy with combination of interferon and ribavirin, and to determine the anti-viral potential of Cyclosporine in vitro. Immunosuppression consisted of either Cyclosporine or Tacrolimus-based therapy. Both groups received therapy with interferon and ribavirin for 48 weeks when evidence of progressive histologic disease was determined. We found that subjects on Cyclosporine-based immunosuppression (n = 56) had a higher sustained virological response of 46% compared to 27% in the patients on Tacrolimus-based therapy (n = 59, P = 0.03). In vitro studies were performed to evaluate the antiviral effect of Cyclosporine in the replicon system. These studies showed that Cyclosporine inhibits hepatitis C viral replication in a dose-dependent manner. Combination of Cyclosporine with interferon showed additive effect, and its function is independent of interferon signaling pathways. In conclusion, Cyclosporine may offer an advantage to Tacrolimus in those patients undergoing interferon-based therapy and should be studied in a prospective randomized trial. PMID- 16382465 TI - Recurrent hepatitis C posttransplant: early preservation injury may predict poor outcome. AB - Organ cold/warm ischemia is thought to be a risk factor for increased severity of recurrence of hepatitis C (HCV) post liver transplantation. We had noted some HCV patients with preservation injury (PI) to have particularly poor outcomes. Our goal was to determine if PI on biopsy in HCV patients is associated with earlier, more rapidly progressive recurrence or graft and patient survival. Sixty-nine patients from the University of Nebraska transplant database were included: 23 HCV patients with PI (group = 1), 23 non-HCV patients with PI (group = 2), and 23 HCV patients without PI (group = 3). Patient groups were matched for gender, age, immunosuppression, and time of transplantation for analysis. No difference in time to recurrence was noted between HCV groups (256 vs. 316 days posttransplant). More patients in group 1 had progression to stage 3 or 4 fibrosis, compared to group 3 (43 vs. 9%, P = 0.02). One-year survival for groups 1, 2, and 3 was 78, 82, and 100% respectively, whereas 3-yr survival was 59, 82, and 88% (group 1 vs. group 2 or 3 respectively, P = 0.0055). There was no difference in survival between groups 2 and 3. Patients in group 1 that received antiviral treatment had improved survival, compared to those who did not (P = 0.012). Risk factors for poor survival on univariate analysis included severity of PI (Relative Risk = 2.78, P < 0.001) and donor age of >55 (P = 0.014). Multivariate analysis shows HCV is the most important factor. In conclusion, HCV transplant patients with evidence of early PI on biopsy have poorer survival outcomes than non-HCV transplant patients with PI or HCV transplant patients without PI. Consideration for antiviral therapy early in the posttransplant course may be warranted in this subset of patients. PMID- 16382466 TI - Fatal graft-versus-host disease after living donor liver transplantation: differential impact of donor-dominant one-way HLA matching. AB - Graft-versus-host disease (GVHD) is an uncommon but potentially devastating complication following liver transplantation. Recently, it was shown that use of a human leukocyte antigen (HLA)-homozygous donor leading to one-way HLA matching significantly increases the risk of GVHD after living donor liver transplantation (LDLT). However, the precise impact of HLA matching between donor and recipient on the risk of GVHD is not yet clear. We surveyed instances of fatal GVHD following LDLT in Japan and reviewed all 8 cases in detail, especially with respect to HLA matching. Serological typing showed that 7 of those cases had donor-dominant one-way HLA matching in the 3 loci of HLA-A, -B, and -DR, while one had donor-dominant one-way HLA matching in the 2 loci of HLA-A and -DR and identical alleles in the B locus. However, DNA typing revealed that the latter case had 1-way HLA matching in the 3 loci. Further, we analyzed HLA typing of 906 donor-recipient pairs who underwent LDLT. There were 5 cases with donor-dominant one-way matching in 2 loci and 2 with donor-dominant one-way matching in 1 locus. All of those cases except 1, who died from an unrelated cause, are alive without an obvious presentation of GVHD. In conclusion, our results suggest that the total number of loci with donor-dominant one-way HLA matching is important for determining the risk of fatal GVHD following LDLT, and that DNA typing of HLA alleles is indispensable in some cases to identify the true risk of donor dominant 1-way HLA matching. PMID- 16382467 TI - Hepatic artery thrombosis following orthotopic liver transplantation: a 10-year experience from a single centre in the United Kingdom. AB - Hepatic artery thrombosis (HAT) occurs in 3-9% of all liver transplants and acute graft loss is a possible sequelae. We present our experience in the management of HAT over a 10-year period. Prospectively collected data from April 1994 to April 2004 were analyzed. There were 1,257 liver transplants, 669 males, median age 51 (16-73) years. There were 61 (4.9%) cases of HAT. Early HAT occurred in 21 (1.8%). Thirty six had graft dysfunction, 11 required a regraft, and 14 died. Positive CMV serology in the donor, cold ischemia time, duration of operation, transfusions of more than 6 units of blood, and 15 units of plasma, an aortic conduit for arterial reconstruction, Roux-en-Y biliary reconstructions, regrafts and relaparotomy were associated with HAT. At multivariate analysis, type of biliary anastomosis was the only significant factor associated with HAT. Split or reduced liver graft were not risk factors for HAT. Number of hepatic arteries requiring multiple arterial anastomosis was not a risk for HAT. HAT resulted in a reduction in overall survival post liver transplantation. The incidence of HAT was 4.9%; with 1.8% early HAT and HAT impacted on survival. Surgical technique was not an aetiological factor for HAT. In conclusion, while a Roux-en-Y biliary reconstruction was an independent risk factor for HAT, cold ischemia and operative times, the use of blood and plasma and the use of aortic conduits in arterial reconstruction were associated with HAT. Regrafts and reoperation were also identified risk factors. PMID- 16382468 TI - Liver laceration associated with severe seizures after living donor liver transplantation. AB - Hemorrhagic complications commonly occur early after liver transplantation (LT), sometimes requiring emergent relaparotomy. However, active bleeding from the liver graft itself is a rare but life-threatening complication after living donor liver transplantation (LDLT). We report an unusual case of liver laceration with massive bleeding, associated with severe epileptic seizures as a result of tacrolimus-induced leukoencephalopathy, after LDLT. The patient was successfully rescued by conventional surgical management without a second transplantation. In conclusion, to our knowledge this is the first reported case of graft rupture due to immunosuppression-associated leukoencephalopathy after LT. PMID- 16382469 TI - Outflow vein reconstruction of extended right lobe graft using quilt venoplasty technique. PMID- 16382470 TI - The MELD score as a predictor of nontransplant surgical mortality in patients with cirrhosis. PMID- 16382471 TI - Histologic characteristics of late cellular rejection, significance of centrilobular injury, and long-term outcome in pediatric liver transplant recipients. AB - Cellular rejection is a common event in orthotopic liver transplantation, leading to significant morbidity and mortality. Late acute cellular rejection, which occurs at least 3 months post-transplant, affects 8-32% of pediatric liver transplant recipients. The histopathology and clinical outcome of patients affected by late cellular rejection are incompletely understood. The aims of this study were 1) to characterize late cellular rejection in the pediatric liver transplant population and describe the histopathology of late cellular rejection, with specific attention to centrilobular injury, including necrosis, inflammation, and endothelialitis and 2) to characterize the long-term outcome of pediatric liver transplant recipients with centrilobular injury, including necrosis, inflammation, and endothelialitis. All liver biopsies performed from August 1997 to August 2002 on pediatric patients at least 6 months post transplant were reviewed, scored for rejection by Banff criteria and examined for centrilobular inflammatory changes. Histology was then correlated with clinical outcomes. Fifteen percent of biopsies had late cellular rejection by Banff criteria. Centrilobular inflammation, necrosis and central vein endothelialitis were common in these biopsies. In patients with similar centrilobular changes, but not meeting Banff criteria for rejection, 29% were treated for rejection and responded well clinically. The long-term outcome of patients with isolated centrilobular injury is similar to that of patients with centrilobular changes associated with portal based rejection. In conclusion, the presence of centrilobular inflammation, necrosis, or central vein endothelialitis should prompt consideration of late cellular rejection. PMID- 16382472 TI - Regarding diarrhea in liver transplant recipients: etiology and management. PMID- 16382473 TI - Proposal of a modified Child-Turcotte-Pugh scoring system and comparison with the model for end-stage liver disease for outcome prediction in patients with cirrhosis. AB - The model for end-stage liver disease (MELD) has a better predictive accuracy for survival than the Child-Turcotte-Pugh (CTP) system and has been the primary reference for organ allocation in liver transplantation. The CTP system, with a score range of 5-15, has a ceiling effect that may compromise its predictive power. In this study, we proposed a refined CTP scoring method and investigated its predictive ability. An additional point was given to patients with serum albumin < 2.3 g/dL, bilirubin > 8 mg/dL or prothrombin time prolongation > 11 seconds. The modified CTP system, containing class D, was compared to the MELD and original CTP system in 436 patients. There was a significant correlation between the MELD and modified CTP score (rho = 0.59, P< 0.001). Using mortality as the endpoint, the area under receiver operating characteristic curve for modified CTP system was 0.895 compared with 0.872 for MELD (P = 0.450) and 0.809 for original CTP system (P < 0.001) at 3 months; the area was 0.890, 0.837 and 0.756, respectively (P = 0.051 and < 0.001, respectively) at 6 months. The risk ratio per unit increase for the modified CTP score was 2.7 and 3.08 at 3 and 6 months respectively (P < 0.001). In conclusion, the modified CTP system can be proposed as an alternative prognostic model for cirrhotic patients. By extending the score range according to the influence of the laboratory-derived variables, the modified CTP system has a better performance than the original system and is as efficient as the MELD for outcome prediction. PMID- 16382474 TI - Navigator-echo-based MR provides high-resolution images and precise volumetry of swine livers without breath holding or injection of contrast media. AB - The accurate calculation of hepatic volume by computed tomography (CT) or magnetic resonance (MR) is complicated by the need for breath holding and the injection of contrast media. These are often contraindicated in patients with liver failure, and we examined the ability of unenhanced 3-dimensional (3-D) navigator-echo-based MR (NE-MR) to accurately image livers and measure volumes without breath holding compared to unenhanced (plain) or gadolinium-diethylene triamine pentaacetic acid enhanced MR (Gd-MR) in miniature swine (n = 8). Without breath holding, diaphragm movement monitoring with NE-MR reduced motion artifacts in hepatic images compared with the other modalities. Without the injection of contrast media, the signal-to-noise ratios of the images obtained using NE-MR were significantly higher than those from plain MR; Gd-MR was superior to NE-MR, however (79.5 +/- 7.5 vs. 63.2 +/- 6.0 or 97.8 +/- 8.1, respectively; P < 0.01 for each). Overall, NE-MR produced improved high-resolution liver images. Consequently, liver volumes calculated based on NE-MR images were more highly correlated with actual liver weights compared to plain or Gd-MR in the whole livers (n = 8; r = 0.937 vs. 0.835 or 0.904, respectively). Also, NE-MR demonstrated significantly strong correlation between actual weights and volumetry-calculated volumes in regenerative livers 7 days after massive hepatectomy (n = 10, r = 0.989, P < 0.01). In conclusion, our results indicate that without breath holding or the injection of contrast media, 3-D NE-MR can provide both high-resolution liver images and precise hepatic volumes in patients with liver failure due to liver surgery (massive hepatectomy and living donor liver transplantation) or fulminant hepatic failure. PMID- 16382475 TI - Preincubation of rat and human hepatocytes with cytoprotectants prior to cryopreservation can improve viability and function upon thawing. AB - Cryopreservation of human hepatocytes is important for the treatment of liver disease by hepatocyte transplantation and also for the use of hepatocytes as an in vitro model of the liver. One factor in the success of cryopreservation is the quality of cells before freezing. Preincubation of hepatocytes with cytoprotective compounds to allow recovery from the isolation process prior to cryopreservation, such as those that will boost cellular adenosine triphosphate (ATP) content or antioxidants, may improve the viability and function of cells upon thawing. Rat hepatocytes were used to investigate the effects of preincubation with 10 compounds: precursors (glucose, fructose, glutathione, and S-adenosyl-L-methionine), antioxidants (ascorbic acid and alpha-lipoic acid), and compounds with multiple effects (N-acetylcysteine, pentoxifylline, prostaglandin E1, and tauroursodeoxycholic acid). Human hepatocytes were then used to investigate 5 of the original 10 compounds (glucose, fructose, alpha-lipoic acid, S-adenosyl-L-methionine, and pentoxifylline). Glucose preincubation (100 - 300 mM) improved the viability and attachment efficiency of rat hepatocytes and improved the viability and reduced lactate dehydrogenase (LDH) leakage of human hepatocytes. Fructose preincubation (100 - 300 mM) improved the viability and attachment efficiency of rat hepatocytes and improved the attachment efficiency of human hepatocytes. alpha-lipoic acid preincubation (0.5 - 5 mM) improved the viability and attachment efficiency of both rat and human hepatocytes. At a concentration of 2.5 mM alpha-lipoic acid also improved the albumin production of human hepatocytes. In conclusion, preincubation of hepatocytes prior to cryopreservation can improve the viability and function of thawed cells and may provide a method of obtaining better-quality cryopreserved hepatocytes for transplantation. PMID- 16382477 TI - Liver repopulation and long-term function of rat small hepatocyte transplantation as an alternative cell source for hepatocyte transplantation. AB - Hepatocyte transplantation (HT) is an attractive therapeutic modality for liver disease as an alternative for liver organ transplantation. Primary fresh hepatocytes (FHs) are the exclusive cell source that has been used for clinical HT. However, the use of FHs is limited due to a shortage of donor cells. Small hepatocytes (SHs) are hepatic progenitor cells and can be isolated not only from rodents but also from humans. SHs can proliferate in vitro and express liver functions, although conventional hepatocytes lose them within a short period after culture. SH functions in vivo have never been studied. We therefore investigated HT using SHs to evaluate cell engraftment and function compared to HT using FHs. The donor cell number in the SH group was smaller than that in the FH group at HT. The cell engraftment in the SH group was smaller in the liver and larger in the spleen than in the FH group. The cell engraftment in the liver increased after HT; however, that in the spleen decreased after HT in both groups. HT using SHs supported the serum albumin level in the NAR experiment as well as that using FH, and albumin mRNA was detectable in the recipients' tissues at 12 weeks after HT. In conclusion, HT using SHs showed hepatic repopulation similar to that using FHs. This suggests that both SHs and FHs can repopulate the liver as if they were hepatic stem cells. In addition, HT using SHs supported liver functions such as albumin correction at the same level as that using FHs. These observations strongly support the idea that SHs could be an alternative to primary FHs as a novel cell source for future HT. PMID- 16382478 TI - Test strips for heavy-metal ions fabricated from nanosized dye compounds. PMID- 16382479 TI - Isostructurality, polymorphism and mechanical properties of some hexahalogenated benzenes: the nature of halogen...halogen interactions. AB - The nature of intermolecular interactions between halogen atoms, X...X (X = Cl, Br, I), continues to be of topical interest because these interactions may be used as design elements in crystal engineering. Hexahalogenated benzenes (C6Cl(6 n)Br(n), C6Cl(6-n)I(n), C6Br(6-n)I(n)) crystallise in two main packing modes, which take the monoclinic space group P2(1)/n and the triclinic space group P1. The former, which is isostructural to C6Cl6, is more common. For molecules that lack inversion symmetry, adoption of this monoclinic structure would necessarily lead to crystallographic disorder. In C6Cl6, the planar molecules form Cl...Cl contacts and also pi...pi stacking interactions. When crystals of C6Cl6 are compressed mechanically along their needle length, that is, [010], a bending deformation takes place, because of the stronger interactions in the stacking direction. Further compression propagates consecutively in a snakelike motion through the crystal, similar to what has been suggested for the motion of dislocations. The bending of C6Cl6 crystals is related to the weakness of the Cl...Cl interactions compared with the stronger pi...pi stacking interactions. The triclinic packing is less common and is restricted to molecules that have a symmetrical (1,3,5- and 2,4,6-) halogen substitution pattern. This packing type is characterised by specific, polarisation-induced X...X interactions that result in threefold-symmetrical X3 synthons, especially when X = I; this leads to a layered pseudohexagonal structure in which successive planar layers are inversion related and stacked so that bumps in one layer fit into the hollows of the next in a space-filling manner. The triclinic crystals shear on application of a mechanical stress only along the plane of deformation. This shearing arises from the sliding of layers against one another. Nonspecificity of the weak interlayer interactions here is demonstrated by the structure of twinned crystals of these compounds. One of the compounds studied (1,3,5-tribromo-2,4,6-triiodobenzene) is dimorphic, adopting both the monoclinic and triclinic structures, and the reasons for polymorphism are suggested. To summarise, both chemical and geometrical models need to be considered for X...X interactions in hexahalogenated benzenes. The X...X interactions in the monoclinic group are nonspecific, whereas in the triclinic group some X...X interactions are anisotropic, chemically specific and crystal-structure directing. PMID- 16382480 TI - An efficient enzymatic synthesis of benzocispentacin and its new six- and seven membered homologues. AB - A very efficient enzymatic method was developed for the synthesis of new enantiomeric benzocispentacin and its six- and seven-membered homologues through the Lipolase (lipase B from Candida antarctica) catalyzed enantioselective (E > 200) ring opening of 3,4-benzo-6-azabicyclo[3.2.0]heptan-7-one, 4,5-benzo-7 azabicyclo[4.2.0]octan-8-one, and 5,6-benzo-8-azabicyclo[5.2.0]nonan-9-one with H2O in iPr2O at 60 degrees C. The (1R,2R)-beta-amino acids (ee > or = 96%, yields > or = 40%) and (1S,6S)-, (1S,7S)-, and (1S,8S)-beta-lactams (ee > 99%, yields > or = 44%) produced could be easily separated. The ring opening of racemic and enantiomeric beta-lactams with 18% HCl afforded the corresponding beta-amino acid hydrochlorides. PMID- 16382481 TI - Effective detection of peptides containing cysteine sulfonic acid using matrix assisted laser desorption/ionization and laser desorption/ionization on porous silicon mass spectrometry. AB - Cysteine sulfonic acid-containing peptides, being typical acidic peptides, exhibit low response in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. In this study, matrix conditions and the effect of diammonium hydrogencitrate (DAHC) as additive were investigated for ionization of cysteine sulfonic acid-containing peptides in MALDI. A matrix-free ionization method, desorption/ionization on porous silicon (DIOS), was also utilized to evaluate the effect of DAHC. When equimolar three-component mixtures of peptides carrying free cysteine, cysteine sulfonic acid, and carbamidomethyl cysteine were measured by MALDI using a common matrix, alpha-cyano-4-hydroxycinnamic acid (CHCA), no signal corresponding to cysteine sulfonic acid-containing peptide could be observed in the mass spectrum. However, by addition of DAHC to CHCA, the peaks of cysteine sulfonic acid-containing peptides were successfully observed, as well as when using 2,4,6-trihydroxyacetophenone (THAP) and 2,6-dihydroxyacetophenone with DAHC. In the DIOS mass spectra of these analytes, the use of DAHC also enhanced the peak intensity of the cysteine sulfonic acid-containing peptides. On the basis of studies with these model peptides, tryptic digests of oxidized peroxiredoxin 6 were examined as a complex peptide mixture by MALDI and DIOS. In MALDI, the peaks of cysteine sulfonic acid-containing peptides were observed when using THAP/DAHC as the matrix, but this was not so with CHCA. In DIOS, the signal from cysteine sulfonic acid-containing peptides was suppressed; however, the use of DAHC significantly enhanced the signal intensity with an increase in the number of observed peptides and increased signal-to-noise ratio in the DIOS spectra. The results show that DAHC in the matrix or on the DIOS chip decreases discrimination and suppression effects in addition to suppressing alkali-adduct ions, which leads to a beneficial effect on protonation of peptides containing cysteine sulfonic acid. PMID- 16382482 TI - Evaluation of the reduction of imidazophenazine dye derivatives under fast-atom bombardment mass-spectrometric conditions. AB - Satellite [M + 2](+*) and [M + 3](+) peaks accompanying the common peak of the protonated molecule [M + H](+) that are known to indicate the occurrence of a reduction process were observed in the fast atom bombardment (FAB) mass spectra of imidazophenazine dye derivatives in glycerol matrix. The distribution of the abundances in the [M + nH](+) peak group varied noticeably for different derivatives. This indicated different levels of the reduction depending on the different structure variations of the studied molecules. In the search for correlations between the mass spectral pattern and the structural features of the dyes, ab initio HF/6-31++G** quantum chemical calculations were performed. They revealed that the abundances of the [M + 2](+*) and [M + 3](+) ions show growth proportional to the decrease of the energy of the lowest unoccupied molecular orbital, i.e. proportional to the increase of the electron affinity of the dye molecule. A method for rapid screening of reductive properties of sets of dye derivatives on the basis of the FAB mass spectral data is discussed. PMID- 16382483 TI - A validated positive chemical ionization GC/MS method for the identification and quantification of amphetamine, opiates, cocaine, and metabolites in human postmortem brain. AB - A sensitive and specific method for the simultaneous detection and quantification of amphetamine, opiates, and cocaine and metabolites in human postmortem brain was developed and validated. Analytes of interest included amphetamine, morphine, codeine, 6-acetylmorphine, cocaine, benzoylecgonine, ecgonine methyl ester, ecgonine ethyl ester, cocaethylene, and anhydroecgonine methyl ester. The method employed ultrasonic homogenization of brain tissue in pH 4.0 sodium acetate buffer and solid phase extraction. Extracts were derivatized with N-methyl-N (tert-butyldimethylsilyl) trifluoroacetamide and N,O-bis(trimethylsilyl) trifluoroacetamide. Separation and quantification were accomplished on a bench top positive chemical ionization capillary gas chromatograph/mass spectrometer with selected ion monitoring. Eight deuterated analogs were used as internal standards. Limits of quantification were 50 ng/g of brain. Calibration curves were linear to 1000 ng/g for anhydroecgonine methyl ester and 6-acetylmorphine, and to 2000 ng/g for all other analytes. Accuracy across the linear range of the assay ranged from 90.2 to 112.2%, and precision, as percent relative standard deviation, was less than 16.6%. Quantification of drug concentrations in brain is a useful research tool in neurobiology and in forensic and postmortem toxicology, identifying the type, relative magnitude, and recency of abused drug exposure. This method will be employed to quantify drug concentrations in human postmortem brain in support of basic and clinical research on the physiologic, biochemical, and behavioral effects of drugs in humans. PMID- 16382484 TI - Characterization of must and wine of six varieties of grapes by direct infusion electrospray ionization mass spectrometry. AB - Samples of must derived from six different varieties of grapes taken during the fermentation process, as well as the respective wine samples directly after the end of the malolactic fermentation, were analyzed by direct infusion negative ion mode electrospray ionization mass spectrometry (ESI-MS). Diagnostic ions for must were different from those of wine samples, although small variations for each of the grape varieties were also detected. The addition of unfermented must or sugar to wine could also be clearly detected. The spectra were acquired in a few minutes per sample, indicating that ESI-MS can be used for high-throughput analysis of samples and should prove useful for quality control during and after the fermentation process. PMID- 16382485 TI - Non-linearity influences on the axial secular-motion spectrum of ions confined in a Fourier transform ion trap mass spectrometer. Experimental studies. AB - A Fourier transform operating mode is applied to an ion trap. The trap is truncated at 2r(0) and presents unwanted defects that induce confinement electric field non-linearities. Ion axial secular-motion spectrum is examined by experiments near the resonance line beta(z) = 0.5. Ion-loss processes and ion axial-motion peak splitting are observed. In the non-linear ion trap, the ion motion frequency depends on its initial conditions in position and velocity. This brings an enlargement of the motion-frequency peak and limits the resolution. With a 2r(0) truncated ion trap, the Fourier transform ion trap mass spectrometer (FTIT-MS) leads experimentally to a mass resolution of about 4000 at 130 u. PMID- 16382486 TI - Rapid sequencing and disulfide mapping of peptides containing disulfide bonds by using 1,5-diaminonaphthalene as a reductive matrix. AB - MS/MS is indispensable for the amino acid sequencing of peptides. However, its use is limited for peptides containing disulfide bonds. We have applied the reducing properties of 1,5-diaminonaphthalene (1,5-DAN) as a MALDI matrix to amino acid sequencing and disulfide bond mapping of human urotensin II possessing one disulfide bond, and human guanylin possessing two disulfide bonds. 1,5-DAN was used in the same manner as the usual MALDI matrices without any pre-treatment of the peptide, and MS/MS was performed using a matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometer (MALDI QIT TOFMS). The results demonstrated that MS/MS of the molecular ions reduced by 1,5-DAN provided a series of significant b-/y-product ions. All 11 amino acid residues of urotensin II were identified using 1,5-DAN, while only 5 out of 11 residues were identified using 2,5-dihydroxybenzoic acid (DHB); similarly 11 out of 15 amino acid residues of guanylin were identified using 1,5-DAN, while only three were identified using DHB. In addition, comparison of the theoretical and measured values of the mass differences between corresponding MS/MS product ions using 1,5-DAN and DHB narrowed down the possible disulfide bond arrangement candidates. Consequently, 1,5-DAN as a reductive matrix facilitates rapid amino acid sequencing and disulfide mapping for peptides containing disulfide bonds. PMID- 16382487 TI - Novel acetylation-aided migrating rearrangement of uridine-diphosphate-N acetylglucosamine in electrospray ionization multistage tandem mass spectrometry. AB - Uridine 5'-diphospho-N-acetylglucosamine (UDP-GlcNAc) is the final product of hexosamine biosynthetic pathway (HSP) and the donor substrate for the modification of nucleocytoplasmic proteins at serine and threonine residues with N-acetylglucosamine (GlcNAc) catalyzed by O-GlcNAc transferase (OGT). Many analogs of UDP-GlcNAc were designed to interfere with the process of protein O glycosylation by blocking OGT. A novel rearrangement reaction was observed in which phosphate-N-acetylglucosamine moiety migrated to 3' terminus of ribose in ESI-MS(n) of UDP-GlcNAc. Results from tandem mass spectrometry, control experiments and calculation showed that the phosphate-N-acetylglucosamine migration might undergo a pentacoordinate phosphoric intermediate. Furthermore, the acetylation of glucosamine in UDP-GlcNAc was essential in the migration process. PMID- 16382488 TI - On the application of the von Mises distribution and angular regression methods to investigate the seasonality of disease onset. AB - This paper describes an approach to summarize the data arising from studies investigating the pattern of disease onset within a calendar year. Such data have been traditionally summarized into monthly counts summated over the complete years studied and patterns often examined by use of Pearson's chi(2) tests with 11 degrees of freedom. This test and others commonly used in practice are reviewed. As an alternative, we suggest that by first representing the date of onset for an individual as a point on a unit circle that the von Mises distribution with a single peak may provide a useful description of such data. Further an extension to angular regression including covariates, analogous to that used routinely in other areas of clinical research, potentially allows a more systematic and detailed investigation of possible seasonal patterns in patient subgroups. The methodology is applied to examples from the date of onset of primary angle-closure glaucoma and date of diagnosis of acute lymphoblastic leukaemia and examines in both situations how the peak onset varies with covariates. Difficulties associated with convergence to the maximum likelihood estimates of the associated parameters are described. Finally, we emphasize the need for individualized (rather than grouped) patient data to be available for study, a clear specification of the particular 'onset' time studied, and suggest that further case studies are required to evaluate the approach. PMID- 16382489 TI - More serious letters about obesity. PMID- 16382490 TI - More serious letters about obesity. PMID- 16382491 TI - More serious letters about obesity. PMID- 16382493 TI - More serious letters about obesity. PMID- 16382492 TI - More serious letters about obesity. PMID- 16382494 TI - More serious letters about obesity. PMID- 16382495 TI - Readers' responses to the webcast video editorials entitled "the American healthcare 'system' in 2005"--parts 1 and 2. PMID- 16382496 TI - Readers' responses to the webcast video editorial entitled "caution--lab test ordering trouble ahead". PMID- 16382497 TI - Readers' responses to the webcast video editorial entitled "caution--lab test ordering trouble ahead". PMID- 16382498 TI - Readers' responses to the webcast video editorial entitled "caution--lab test ordering trouble ahead". PMID- 16382499 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382500 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382501 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382502 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382503 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382504 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382505 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382506 TI - Readers' responses to "help wanted: more doctors for the United States". PMID- 16382507 TI - Diet, nutrition, and the life-course approach to cancer prevention. AB - Cancer results from the interaction of genetic susceptibility and environmental exposures. The diagnosis of cancer is age related; there is a marked increase in cancer incidence after the reproductive years. Nutrient and toxicant exposures are important contributors to the risk of some cancers. Nutrition, as a determinant of growth and body composition, also influences cancer risk, directly due to carcinogens in foods or indirectly by the hormonal and metabolic response to growth and obesity. There is strong evidence that obesity and rapid growth enhance the risk of cancer. The prevention of cancer should start before conception; mothers should start pregnancy with a healthy weight and avoid excessive or low weight gain during pregnancy. Key micronutrients are important for normal embryonic development and fetal growth. Infant growth should be assessed based on optimal health across all stages of the life course, rather than following the present approach of "bigger is better. " This model may increase cancer risk in later life, because bigger is closely linked to fatter. Recent studies of energy expenditure in children indicate that excess energy intakes may have been recommended over the past decades, contributing to the surge in global obesity. Food preferences and habits regarding physical activity and play become set relatively early in life; parents and teachers provide key guidance leading to the adoption of a healthy or an unhealthy lifestyle. Thus, cancer prevention efforts should begin with childhood and continue through all stages of the life course. PMID- 16382508 TI - [Reader's letter concerning the article by H. Strunk et al. (Fortschr Rontgenstr 2005, 177:900-904). Erroneous diagnosis of liver metastasis as benign tumor in the contrast media aided "Low-MI-Real-Time" sonography with SonoVue]. PMID- 16382509 TI - Modafinil for fatigue in MS: a randomized placebo-controlled double-blind study. PMID- 16382510 TI - The rebirth of adult IO. PMID- 16382511 TI - Rescue access made easy. PMID- 16382512 TI - Clinical evaluation of a novel intraosseous device for adults: prospective, 250 patient, multi-center trial. PMID- 16382514 TI - Readers' responses to the webcast video editorial entitled "evidence-based medicine or faith-based medicine?". PMID- 16382513 TI - Readers' responses to the webcast video editorial entitled "evidence-based medicine or faith-based medicine?". PMID- 16382515 TI - Powered intraosseous access in the prehospital setting: MCHD EMS puts the EZ-IO to the test. PMID- 16382516 TI - Adult IO in the combat zone: the past, present and future use of intraosseous infusion by the U.S. military. PMID- 16382517 TI - Profiles in leadership [interview by Teresa McCallion and Keri Losavio]. PMID- 16382518 TI - Intraosseous infusion: bibliography. PMID- 16382519 TI - EMS in Italy. PMID- 16382520 TI - Readers' responses to the webcast video editorial entitled "Willie Sutton and screening for alcohol in the emergency department and trauma center". PMID- 16382521 TI - Readers' responses to the webcast video editorial entitled "Willie Sutton and screening for alcohol in the emergency department and trauma center". PMID- 16382522 TI - Readers' responses to the webcast video editorial entitled "Willie Sutton and screening for alcohol in the emergency department and trauma center". PMID- 16382523 TI - The body in sync: understanding perfusion, fluid balance and all things cardiac. PMID- 16382524 TI - Readers' responses to the webcast video editorial entitled "the official ethics of the American medical profession". PMID- 16382525 TI - Readers' responses to the webcast video editorial entitled "the official ethics of the American medical profession". PMID- 16382526 TI - Readers' responses to the webcast video editorial entitled "the official ethics of the American medical profession". PMID- 16382527 TI - Readers' positive responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382528 TI - Readers' positive responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382529 TI - Readers' positive responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382530 TI - Readers' positive responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382531 TI - Readers' positive responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382532 TI - Readers' negative or neutral responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382533 TI - Readers' negative or neutral responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382534 TI - Readers' negative or neutral responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382535 TI - Readers' negative or neutral responses to the webcast video editorial entitled "how to prevent the obese from becoming 'obeser'--stop eating". PMID- 16382536 TI - Readers' responses to the article entitled "anticoagulation therapy for venous thromboembolism". PMID- 16382537 TI - Readers' responses to the article entitled "anticoagulation therapy for venous thromboembolism". PMID- 16382538 TI - A single-blind, open-label trial of sodium oxybate for myoclonus and essential tremor. AB - The authors performed an open-label, rater-blinded, add-on study of sodium oxybate in 20 patients with ethanol-responsive myoclonus or essential tremor. Blinded ratings of videotaped examinations showed improvements in myoclonus at rest, stimulus-sensitive myoclonus, action myoclonus, functional performance, and postural and kinetic tremor. Tolerability was acceptable, and more than half of the patients chose to continue treatment after the trial. Double-blind placebo controlled studies in these disorders are warranted. PMID- 16382539 TI - Legislation would regulate puppy mills. PMID- 16382540 TI - AVMA Veterinary Medical Assistance Teams. PMID- 16382541 TI - Comparing prescription drug plans. New choices made available by Medicare Part D deserve serious study. PMID- 16382542 TI - Trust has a novel take on scholarships. New England fund exists to help local veterinary students. PMID- 16382543 TI - An American veterinarian in Israel. PMID- 16382544 TI - Questions statistical analysis in equine study. PMID- 16382545 TI - Recommends tattoos for spayed females. PMID- 16382546 TI - Autoantibodies in prostate cancer. PMID- 16382547 TI - Autoantibodies in prostate cancer. PMID- 16382548 TI - Autoantibodies in prostate cancer. PMID- 16382549 TI - Passive immunization against cytomegalovirus during pregnancy. PMID- 16382550 TI - Passive immunization against cytomegalovirus during pregnancy. PMID- 16382551 TI - Chaperones and disease. PMID- 16382552 TI - Chaperones and disease. PMID- 16382554 TI - Management of the lens capsule in pediatric cataract surgery. PMID- 16382553 TI - Pulmonary mucormycosis (zygomycosis) in a lung transplant recipient: recovery after posaconazole therapy. PMID- 16382555 TI - What's your diagnosis? Sporadic nanophthalmos (posterior microphthalmos), bilateral ametropic amblyopia, and esotropia. PMID- 16382556 TI - Newborn primary congenital glaucoma. PMID- 16382557 TI - Newborn primary congenital glaucoma: 2005 update. AB - BACKGROUND: Newborn primary congenital glaucoma is an unusual and important subtype of primary congenital glaucoma. Affected patients typically manifest specific signs that facilitate its recognition at birth and become important in the assessment and determination of appropriate treatment for this severe expression of primary congenital glaucoma. PATIENTS AND METHODS: The medical records of 35 patients with newborn primary congenital glaucoma were reviewed to determine its clinical manifestations and response to therapy. The published literature related to newborn primary congenital glaucoma was reviewed and referenced. RESULTS: Newborn primary congenital glaucoma is recognized at birth because of the associated corneal opacification. Ocular examination reveals anterior segment abnormalities of the cornea, iris, and filtration angle as well as related elevated intraocular pressure. Genetic analysis of a subset of patients with newborn primary congenital glaucoma confirmed its relationship with the less severe infantile form of primary congenital glaucoma, which is characterized by favorable results after goniosurgery. In contrast, goniosurgery was found to have unfavorable results and be less successful compared with trabeculectomy and glaucoma drainage tube shunts as initial therapy for newborn primary congenital glaucoma. CONCLUSIONS: Newborn primary congenital glaucoma is an important subtype of primary congenital glaucoma. It can be differentiated from the more frequent and familiar infantile type by history and careful anterior segment examinations to enable and support appropriate choices for its successful surgical treatment. PMID- 16382558 TI - Acute onset concomitant esotropia: sensorial evaluation, prism adaptation test, and surgery planning. AB - BACKGROUND AND OBJECTIVE: Acute acquired concomitant esotropia is a rare form of strabismus that usually presents during infancy or early childhood with diplopia and minimal refractive error. The aim of this study was to evaluate the response to the prism adaptation test in a group of normosensorial patients with acute acquired concomitant esotropia and to determine its predictive value for surgical outcome. PATIENTS AND METHODS: Six patients (median age, 11 .5 years) with acute acquired concomitant esotropia underwent the prism adaptation test by applications of Fresnel press-on and prismatic lenses. The sensorial condition of each patient had been evaluated with theTNO stereopsis test, the Irvine test, Bagolini striated glasses, and the Worth four-dot test during the prism adaptation test, when maximum compensation angle was achieved, and after surgery. In all patients, surgery was performed based on the angle of the squint after the prism adaptation test. Patients were observed 1 day, 3 months, 6 months, and 1 year after surgery. RESULTS: An increase in the angle of the squint occurred in all patients during prism adaptation. The maximum value was obtained in the same amount of time whether with prisms of increasing power or by direct application of the final value of the prisms. The final prismatic value (target angle) was always the same. CONCLUSIONS: The accuracy of the patients' selection confirmed the existence of highly reproducible compensational movements to the prismatic correction in not only patients with anomalous retinal correspondence but also normosensorial patients. Prism adaptation test results were useful in these patients because of their high predictive value for the surgical treatment. PMID- 16382559 TI - Ocular injuries in children after major trauma. AB - OBJECTIVE: To determine the variety of ocular injuries sustained by children during major trauma. SUBJECTS AND METHODS: The pediatric trauma registry of The Children's Hospital in Denver, Colorado, was surveyed for children with ocular injuries and an Injury Severity Score higher than 15. The injuries were tabulated and correlated with mechanisms of injury to determine identifiable injury patterns. RESULTS: In a pediatric trauma center, ocular injuries were nearly twice as frequent among children with major trauma (Injury Severity Score > 15). Two-thirds of the children with an Injury Severity Score higher than 15 had been involved in a mishap with a motorized or a nonmotorized vehicle. Ocular injuries in children with an Injury Severity Score higher than 15 were characterized by fracture of the surrounding bony structures and contusions. Children with an Injury Severity Score higher than 15 had 50% to 80% fewer open wounds of the ocular adnexa and eyeball than did children with an Injury Severity Score of 15 or lower. The one optic nerve injury and two-thirds of the other ocular cranial nerve injuries occurred in children with an Injury Severity Score higher than 15. CONCLUSIONS: Serious ocular injury should be suspected in children involved in a motor vehicle accident who have an Injury Severity Score higher than 15 and fractures of the surrounding bony structures, contusions, or both. Absence of an open wound of the ocular adnexa or eyeball does not eliminate the possibility of serious ocular injury. PMID- 16382561 TI - Randot stereoacuity testing in young children. AB - OBJECTIVE: The Randot test is a commonly used stereoacuity test with child friendly targets; however, little has been published about its use for young children. We sought to determine the completion rate of the Randot test among young children with normal vision and to calculate normative values. SUBJECTS AND METHODS: The Randot stereoacuity test was administered to 264 children whose ages ranged from 3 to 7 years. Because this was an investigation to determine the percentage of children who could complete the Randot test, children who failed a vision screening modified from the Modified Clinical Technique were excluded to minimize confounding factors (ie, a lack of measurable stereopsis due to vision problems rather than inability to understand the test). Completion rates and cumulative percentages were calculated by age. RESULTS: A completion rate of 98.5% was found. Median Randot stereoacuities were 100 seconds of arc for the 3 year-old children, 70 seconds of arc for the 4-year-old children, 50 seconds of arc for the 5-year-old children, 40 seconds of arc for the 6-year-old children, and 45 seconds of arc for the 7-year-old children. Kruskal-Wallis testing revealed a significant change in stereoacuity with age (chi-square, 35.55; df = 4; P < .001), and pairwise comparisons (alpha = 0.05) revealed that the 3- and 4 year-old children were significantly different from the 5-, 6-, and 7-year-old children. CONCLUSIONS: Randot stereoacuity can be assessed for most preschool children and all young school-aged children. In the absence of significant refractive error, strabismus, or reduced or unequal acuity, most 4-year-old children should have a Randot stereoacuity of 70 seconds of arc or better and most young school-aged children should have a Randot stereoacuity of 50 seconds of arc. PMID- 16382560 TI - Evaluation of intranasal midazolam in refraction and fundus examination of young children with strabismus. AB - PURPOSE: To determine the clinical sedative effect and dosage of intranasal midazolam in refraction and fundus examination of children with strabismus. PATIENTS AND METHODS: Refraction and fundus examination with (n = 28) and without (n = 24) sedation were performed in 52 children with strabismus whose ages ranged from 7 to 26 months. We delivered a 5-mg/mL solution of midazolam via a syringe for 60 seconds to provide a dose of 0.2 mg/kg. We repeated the dose to a maximum of 0.3 mg/kg if there was no clinical sedative response after 10 to 15 minutes. The ease of examination and sedation for each patient was scored by a blinded observer. The groups were evaluated for ease of examination and the time needed to complete it. RESULTS: We obtained clinically adequate sedation at a mean (+/- standard deviation) of 15 minutes (+/- 2.69 minutes). Sedation was achieved with a mean dose of 2.64 mg/kg (+/- 0.66 mg/kg). Children receiving midazolam had significantly calmer examination scores. The time needed to complete the examination was statistically significantly shorter for these children than for children not receiving sedation (P < .05). CONCLUSIONS: Intranasal midazolam is a beneficial drug and method of delivery for the sedation of anxious children with strabismus undergoing refraction and fundus examination. Sedation prior to examination is effective in reducing the anxiety and time associated with ophthalmologic examination of children with strabismus. PMID- 16382562 TI - Laser treatment of Coats' disease. AB - A 9-year-old boy presented with an exudative retinal detachment, vascular telangiectasias, subretinal lipid, and retinal macrocyst formation. He underwent three sessions of large-spot diode laser for the treatment of Coats' disease. Serial examinations and fundus photography documented an excellent involutional response with an improvement in visual acuity. PMID- 16382563 TI - Orbital hydatid cyst manifested as expanding exophthalmos following blunt orbital trauma. AB - A 14-year-old boy suffered right blunt orbital trauma. X-ray revealed a compressed fracture of the right ethmoidal sinuses. Three months after the trauma, progressive, painless right proptosis developed. Ocular examination revealed a severe proptosis and pronounced down displacement of the right globe without signs of orbital inflammation. A well-demarcated, extraconal right orbital cystic mass was seen on computerized tomography scan. The cyst contents were aspirated and found to contain echinococcal scolices and "hydatid sand." Hypertonic saline was injected and the cyst was excised. Progressive proptosis after blunt orbital trauma in patients from endemic areas should be suspected of being an orbital hydatid cyst. PMID- 16382564 TI - A case of hypopyon uveitis nonresponsive to steroid therapy and a review of anterior segment masquerade syndromes in childhood. AB - We report a case of hypopyon uveitis referred as inflammatory disease nonresponsive to corticosteroids. A detailed history revealed that the patient had been treated previously for acute lymphoblastic leukemia (ALL). Anterior chamber tap was consistent with recurrence of ALL. A review of pseudouveitis in the pediatric population is also presented. PMID- 16382565 TI - Lamellar macular hole as the presenting feature in a child with Coats' disease. AB - We report a case of Coats' disease and lamellar macular hole in a 10-year-old boy who presented with blurring of vision in his left eye. This is the first reported case of Coats' disease presenting with a lamellar macular hole in a child. PMID- 16382566 TI - Traumatic iridodialysis. PMID- 16382567 TI - Pharmacotherapeutic options in pain management. PMID- 16382568 TI - Managing persistent neuropathic pain in the elderly. PMID- 16382569 TI - Recent advances in the biochemistry and clinical relevance of the isoprostane pathway. AB - Isoprostanes (IsoPs), lipid peroxidation products formed via the free radical mediated oxidation of arachidonic acid, have become the "gold standard" biomarker of oxidative stress in vivo over the past 15 yr. Significant advances have been made in understanding this important pathway of lipid peroxidation. Recent studies from our laboratory are discussed that have provided insights into the mechanism of formation and regioisomeric distribution of these compounds and that have identified novel products of the IsoP pathway such as cyclized dioxolane IsoPs, IsoP-derived racemic prostaglandins, and reactive cyclopentenone IsoP, the latter of which possess potent biological actions. Furthermore, new independent studies have demonstrated that IsoPs are the most reliable available marker of lipid peroxidation in vivo, and recent work examining IsoP formation has provided valuable information about the pathogenesis of numerous human diseases. Thus, the complexity of the IsoP pathway has expanded, providing novel insights into mechanisms of lipid peroxidation in vivo and allowing investigators to explore the role of oxidative stress in human disease. PMID- 16382570 TI - Paradoxical effect of n-3-containing vegetable oils on long-chain n-3 fatty acids in rat heart. AB - Flaxseed, echium, and canola oils contain alpha-linolenic acid (18:3n-3, ALA) in a range of concentrations. To examine their effect on elevating cardiac levels of long-chain n-3 FA, diets based on these n-3-containing vegetable oils were fed to rats for 4 wk. Sunflower oil, which contains little ALA, was a comparator. Despite canola oil having the lowest ALA content of the three n-3-containing vegetable oils, it was the most potent for elevating DHA (22:6n-3) levels in rat hearts and plasma. However, the relative potencies of the dietary oils for elevation of EPA (20:5n-3) in heart and plasma followed the same rank order as their ALA content, i.e., flaxseed > echium > canola > sunflower oil. This paradox may be explained by lower ALA intake leading to decreased competition for Delta6 desaturase activity between ALA and the 24:5n-3 FA precursor to DHA formation. PMID- 16382571 TI - Microtubular integrity differentially modifies the saturated and unsaturated fatty acid metabolism in cultured Hep G2 human hepatoma cells. AB - The influence of cytoskeleton integrity on the metabolism of saturated and unsaturated FA was studied in surface cultures and cell suspensions of human Hep G2 hepatoma cells. We found that colchicine (COL), nocodazol, and vinblastin produced a significant inhibition in the incorporation of labeled saturated FA, whereas incorporation of the unsaturated FA remained unaltered. These microtubule disrupting drugs also diminished Delta9-, Delta5-, and Delta6-desaturase capacities. The effects produced by COL were dose (0-50 microM) and time (0-300 min) dependent, and were antagonized by stabilizing agents (phalloidin and DMSO). Dihydrocytochalasin B (20 microM) was tested as a microfilament-disrupting drug and produced no changes in either the incorporation of [14C] FA or the desaturase conversion of the substrates. We hypothesized that the interactions between cytoskeleton and membrane proteins such as FA desaturases may explain the functional organization, facilitating both substrate channeling and regulation of unsaturated FA biosynthesis. PMID- 16382572 TI - Modulation of respiratory syncytial virus-induced prostaglandin E2 production by n-3 long-chain polyunsaturated fatty acids in human respiratory epithelium. AB - Infection with respiratory syncytial virus (RSV) results in substantial infant morbidity and has been associated with the subsequent development of childhood asthma. Inflammatory mediators produced by both the epithelium and tissue leukocytes during RSV infection stimulate the release of chemotactic factors by the respiratory epithelium and the subsequent influx of inflammatory cells, predominantly neutrophils. We investigated the production of inflammatory mediators [prostaglandin E2 (PGE2), interleukin (IL)-1beta, tumor necrosis factor alpha] and chemokines [IL-8, RANTES (regulation on activation, normal T cell expressed and secreted)] by alveolar epithelial cells in response to RSV infection. Infection of a human alveolar epithelial transformed cell line (A549 cells) with live RSV substantially increased production of PGE2, IL-8, and RANTES. By altering cell membrane FA through incorporation of the long-chain PUFA (LCPUFA) arachidonic acid, EPA, and DHA, we were subsequently able to significantly modulate PGE2 production by the infected epithelium. Because of the dynamic nature of the effects of PGE2 on lung function, regulation of this prostaglandin during RSV infection by n-3 LCPUFA has the potential to significantly alter the disease process. PMID- 16382574 TI - A molecular dynamics study of an archaeal tetraether lipid membrane: comparison with a dipalmitoylphosphatidylcholine lipid bilayer. AB - Molecular dynamics simulations of an archaeal membrane made up of bipolar tetraether lipids and a dipalmitoylphosphatidylcholine (DPPC) lipid membrane were performed and compared for the first time. The simulated archaeal membrane consists of a pure monolayer of asymmetrical lipids, analogous to the main polar lipid [MPL; Swain, M., Brisson, J.-R., Sprott, G.D., Cooper, F.P., and Patel, G.B., (1997) Identification of beta-L-Gulose as the Sugar Moiety of the Main Polar Lipid of Thermoplasma acidophilum, Biochim. Biophys. Acta 1345, 56-64] found in T. acidophilum, an extremophile archaeal organism. This simulated membrane lipid contains two cyclopentane rings located on one of the two aliphatic chains of the lipid. The archaeal membrane is simulated at 62degreesC, slightly above the optimal growth temperature of T. acidophilum. We compared the organization of this tetraether lipid monolayer with a DPPC bilayer simulated at 50degreesC, both of them being modeled in a partially hydrated state. Our results assess the singularity of the tetraether lipid organization, in particular the influence of the spanning structure on the molecular ordering within the archaeal membrane. PMID- 16382573 TI - Use of a 13C tracer to investigate lutein as a ligand for plasma transthyretin in humans. AB - The selective accumulation of lutein in the macula of the human retina is likely to be mediated by specific transport and/or binding proteins. Our objective was to determine whether transthyretin (TTR) is a plasma transport protein for lutein. We used a biosynthetic 13C-lutein tracer and GC-combustion interfaced isotope ratio MS to gain the requisite sensitivity to detect the minute amounts of lutein expected as a physiological ligand for TTR. Subjects (n = 4) each ingested 1 mg of 13C-lutein daily for 3 d and donated blood 24 h after the final dose. For three subjects, the plasma TTR-retinol-binding protein (RBP) complex was partially purified by anion-exchange (diethylaminoethyl, DEAE) chromatography and then dissociated by hydrophobic-interaction chromatography to yield the TTR component. For subject 4, the initial DEAE purification step was omitted and total plasma TTR (RBP-bound and free) was isolated by hydrophobic-interaction chromatography. In each case, the crude TTR fractions were then purified to homogeneity by RBP-Sepharose affinity chromatography. Pure TTR was extracted with chloroform, and unlabeled lutein was added to the extract as a carrier. The mean 13C/12C ratio (expressed in delta notation, delta13C) of the lutein fraction isolated from the plasma TTR extracts of the four subjects was -30.53 +/- 3.29 per thousand. The delta13C value of the unlabeled lutein carrier was -30.97 +/- 0.27per thousand. Thus, no 13C enrichment was detected in association with TTR. We conclude that lutein is not associated with TTR in human plasma after being ingested in physiological amounts. PMID- 16382575 TI - 1H and 13C NMR characterization and stereochemical assignments of bile acids in aqueous media. AB - The unconjugated bile acids cholic acid, deoxycholic acid, and chenodeoxycholic acid; their glycine and taurine conjugates glycocholic acid, glycodeoxycholic acid, glycochenodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, and taurochenodeoxycholic acid; and a taurine conjugated ursodeoxycholic acid, tauroursodeoxycholic acid, were characterized through 1H and 13C NMR in aqueous media under the physiological pH region (7.4 +/- 0.1). Assignments of 1H and 13C signals of all the bile acids were made using a combination of several one- and two-dimensional, homonuclear (1H-1H) and heteronuclear (1H-13C) correlations as well as spectral editing NMR methods. Stereochemical assignment of the five membered ring of the bile acids is reported here for the first time. The complete characterization of various bile acids in aqueous media presented here may have implications in the study of the pathophysiology of biliary diseases through human biliary fluids using NMR spectroscopy. PMID- 16382576 TI - Storage lipid accumulation and acyltransferase action in developing flaxseed. AB - Investigations of storage lipid synthesis in developing flaxseed (Linum usitatissimum) provide useful information for designing strategies to enhance the oil content and nutritional value of this crop. Lipid content and changes in the FA composition during seed development were examined in two cultivars of flax (AC Emerson and Vimy). The oil content on a dry weight basis increased steadily until about 20 d after flowering (DAF). The proportion of alpha-linolenic acid (alpha 18:3, 18:3cisDelta9,12,15) in TAG increased during seed development in both cultivars while the proportions of linoleic acid (18:2cisDelta9,12) and saturated FA decreased. The developmental and substrate specificity characteristics of microsomal DAG acyltransferase (DGAT, EC 2.3.1.20) and lysophosphatidic acid acyltransferase (LPAAT, EC 2.3.1.51) were examined using cultivar AC Emerson. The maximal acyltransferase specific activities occurred in the range of 8-14 DAF, during rapid lipid accumulation on a per seed basis. Acyl-CoA of EPA (20:5cisDelta5,8,11,14,17) or DHA (22:6 cis4,7,10,13,16,19) were included in the specificity studies. DGAT displayed enhanced specificity for alpha-18:3-CoA, whereas the preferred substrate of [PAAT was 18:2-CoA. Both enzymes could use EPA or DHA-CoA to varying extents. Developing flax embryos were able to take up and incorporate these nutritional FA into TAG and other intermediates in the TAG formation pathway. This study suggests that if the appropriate acyl-CoA-dependent desaturation/elongation pathways are introduced and efficiently expressed in flax, this may lead to the conversion of alpha-18:3-CoA into EPA-CoA, thereby providing an activated substrate for TAG formation. PMID- 16382577 TI - Cyanolipid-rich seed oils from Allophylus natalensis and A. dregeanus. AB - As a continuation of our study on plants of the Sapindaceae, the chemical composition of the oil extracted from seeds of Allophylus natalensis (Sonder) De Winter and of A. dregeanus (Sonder) De Winter has been investigated. The oil from both species contained approximately equal amounts of TAG and type I cyanolipids (CL), 1-cyano-2-hydroxymethylprop-2-en-1-ol-diesters, with minor amounts of type III CL, 1-cyano-2-hydroxymethylprop-1-en-3-ol-diesters. Structural investigation of the oil components was accomplished by chemical, chromatographic (TLC, CC, GC, and GC-MS), and spectroscopic (IR, NMR) means. GC and GC-MS analysis showed that C20 FA were dominant in the CL components of the oil from the two species (44-80% vs. 21-26% in TAG), with cis-11-eicosenoic acid (36-46%) and cis 13-eicosenoic acid (paullinic acid, 23-37%) as the major esterified fatty acyl chains in A. natalensis and A. dregeanus, respectively. cis-Vaccenic acid was particularly abundant (11-31%) in the CL from A. dregeanus, whereas eicosanoic acid (10-22%) was also a major component of CL in both species. PMID- 16382578 TI - Lipid class distribution of fatty acids including conjugated linoleic acids in healthy and cancerous parts of human kidneys. AB - In this study the FA compositions of healthy and cancerous human renal tissues from the same patients are compared with special reference to the CLA and PUFA content. CLA was preferentially incorporated into neutral lipid compared with phospholipid classes. Its distribution profile was similar to that of monounsaturated FA, but unlike that found with 18:2n-6. Different incorporation patterns were found for individual CLA isomers. Comparing renal cell carcinoma (RCC) and healthy kidney, the total CLA content was significantly lower in the cholesterylester fraction and significantly higher in the PE and PS fractions from RCC. The most significant differences between healthy and cancerous renal tissue were in the content of t10,c12-CLA. Furthermore, the lipid class distributions of n-6 PUFA were determined, and several significant differences between RCC and healthy renal tissue were found. This is of interest, as it has been proposed that the anticarcinogenic properties of dietary CLA are associated with their interference in the metabolism of 20:4n-6. The involvement of CLA in preventing renal cancer cannot be definitively demonstrated from the design of this study, nor was it intended, but the complete determination of the FA composition of adjacent healthy and cancerous tissues may provide an insight if lipids are involved in this disease. PMID- 16382579 TI - Synthesis of a novel series of 2-methylsulfanyl fatty acids and their toxicity on the human K-562 and U-937 leukemia cell lines. AB - The hitherto unknown 2-methylsulfanyldecanoic acid and 2-methylsulfanyldodecanoic acid were synthesized from methyl decanoate and methyl dodecanoate, respectively, through the reaction of lithium diisopropylamide and dimethyldisulfide in THF followed by saponification with potassium hydroxide in ethanol. Both alpha methylsulfanylated FA were cytotoxic to the human chronic myelogenous leukemia K 562 and the human histiocytic lymphoma U-937 cell lines with EC50 values in the 200-300 microM range, which makes them more cytotoxic to these cell lines than decanoic and/or dodecanoic acid. The cytotoxicity of the studied FA toward K-562 followed the order 2-SCH3-12:0 > 2-SCH3-10:0 > 10:0 > 12:0 > 2-OCH3-12:0, whereas toward U-937 the cytotoxicity was 2-SCH3-10:0 > 2-SCH3-12:0 > 12:0 > 10:0 > 2 OCH3-12:0. These results indicate that the alpha-methylsulfanyl substitution increases the cytotoxicity of the C10 and C12 FA toward the studied leukemia cell lines. PMID- 16382580 TI - Kinetics of 14C distribution after tracer dose of 14C-lutein in an adult woman. AB - Lutein is an oxygenated carotenoid (xanthophyll) found in dark green leafy vegetables. High intakes of lutein may lower the risk of age-related macular degeneration. Current understanding of human lutein metabolism as it might occur in vivo is incomplete. Therefore, we conducted a feasibility study where we dosed a normal adult woman with 14C-lutein (125 nmol, 36 nCi 14C), dissolved in olive oil (0.5 g/kg body weight) and mixed in a banana shake. Blood, urine, and feces collected before the dose was administered served to establish baseline values. Thereafter, blood was collected for 63 d following the dose, while feces and urine were collected for 2 wk post-dose. The 14C contents in plasma, urine, and feces were measured by accelerator MS. The 14C first appeared in plasma 1 h after dosing and reached its highest level, approximately2.08% of dose/L plasma, at 14 h post-dose. The plasma pattern of 14C did not include a chylomicrons/VLDL (intestinal) peak like that when the same subject received 14C-beta-carotene (a previous test), suggesting that lutein was handled differently from beta-carotene by plasma lipoproteins. Lutein had an elimination half-life (t1/2) of approximately10 d. Forty-five percent of the dose of 14C was eliminated in feces and 10% in urine in the first 2 d after dosing. Quantifying human lutein metabolism is a fertile area for future research. PMID- 16382581 TI - New ceramides from Rantherium suaveolens. AB - A mixture of five new ceramides was isolated from the aerial parts of Rantherium suaveolens and characterized by spectroscopic and chemical methods. Their structures were elucidated by spectroscopic and chemical methods as (2S,3S, 4R,2'R, 14E)-2-(2'-hydroxydocosanoylamino) - 14 - octadecene -1,3,4-triol (1), (2S,3S,4R,2'R, 14E)-2-(2'-hydroxytricosanoylamino)-14-octadecene-1,3,4-triol (2), (2S,3S,4R,2'R, 14E)-2-(2'-hydroxytetracosanoylamino)- 14 - octadecene - 1,3,4 - triol (3), (2S, 3S,4R,2'R, 14E) - 2 - (2'- hydroxypentacosanoylamino) - 14 - octadecene - 1,3,4-triol (4), and (2S,3S,4R,2'R,14E)-2-(2' hydroxyhexacosanoylamino)-14-octadecene-1,3,4-triol (5). PMID- 16382582 TI - Katrina teaches hard lessons. PMID- 16382583 TI - Ripples in the pond: building a career of influence. PMID- 16382584 TI - Choosing a management firm for STN. PMID- 16382585 TI - A plan to improve end-of-life care for trauma victims and their families. AB - End-of-life care for trauma patients is unique in that clinicians rarely have the advantage of knowing victims prior to the event. In this setting, the unfamiliarity with patients' wishes and values, the critical nature of the injury, the overwhelming feelings of guilt that families often experience, the suddenness and acuity of the crisis, and the need to make life-and-death decisions, can result in end-of-life care scenarios that are challenging to manage and often difficult to experience. This article describes a plan to develop, implement, and test a best practice model of end-of-life care for trauma victims and their families. PMID- 16382586 TI - Making GCS as easy as 1, 2, 3, 4, 5, 6. AB - The Glasgow Coma Scale (GCS) is an easy to use tool for assessing neurological function and brain injury in comatose patients particularly in acute stages of traumatic injury or illness. Due to the simplicity of the scale, however, proper training is often overlooked limiting its usefulness. This manuscript describes the basic components of the GCS and the proper scoring method to elicit accurate evaluations. PMID- 16382587 TI - Update on the National Patient Safety Goals - changes for 2006. AB - Compliance with the NPSGs is mandatory for an organization that seeks JCAHO accreditation. Beyond compliance with goals simply for survey purposes, hospitals that provide care to trauma patients should attempt to make the care of the patient as safe as possible by anticipating problems and complications, and avoiding them if possible. PMID- 16382588 TI - Effectiveness of composite cure associated with different light-curing regimes. AB - This study investigated the use of various light-curing regimens with standardized light energy density on the effectiveness of cure of a visible light activated resin composite (Z100, 3M-ESPE). A light-cure unit (Variable Intensity Polymerizer (VIP), BISCO Inc) which permitted individual control over time and intensity, was used. The five light-curing modes investigated include Pulse Delay (PD), Pulse Cure (PC), Soft-start (SS), Turbo (T) and Control (C). Effectiveness of cure was established by measuring the top and bottom Knoop hardness of 2-mm thick composite specimens using a digital microhardness tester (n=5, load=500g; dwell time=15 seconds) immediately and at one-day post-polymerization. Data obtained was analyzed using one-way ANOVA/Scheffe's post hoc test and Independent Samples t-tests (p<0.05). Top KHN observed immediately after polymerization with C was significantly lower than PD. At one day post-polymerization, the top KHN obtained with C was significantly lower than PD, SS and T. No significant difference in bottom KHN was observed among the different curing modes immediately after curing. At one day post-polymerization, the bottom KHN obtained with C was significantly lower than SS and T. Regardless of curing regimens, top and bottom values at one day were significantly higher than those observed immediately after light polymerization. No significant difference in mean hardness ratio was observed among the different curing regimens immediately and one day later. Effectiveness of the cure at the bottom surfaces of composites may be increased by soft-start and turbo polymerization regimens. PMID- 16382589 TI - Salivary contamination and bond strength of glass-ionomers to dentin. AB - This study evaluated the effect of salivary contamination on the shear bond strength of two highly viscous glass ionomer cements (Fuji IX GP Fast and Ketac Molar Maxicap) to conditioned dentin and assessed the effect of cleaning the contaminated field prior to bonding. The buccal surfaces of 90 human molars and premolars were ground to expose dentin and the teeth were then set in resin. The specimens were divided into two groups for each material, then further subdivided into three groups of 15 teeth each: Group 1--uncontaminated (control), Group 2- dentin contaminated with saliva, Group 3--dentin contaminated, washed and air dried. The specimens were made by bonding the test material to dentin using a 4 mm diameter gelatin capsule. All specimens were protected with varnish and placed in distilled water at 37 degrees C for seven days prior to measuring bond strength in shear. Fractured surfaces were examined visually and by using SEM to assess mode of failure. There were no significant differences in mean shear bond strength among the three test groups for either material (ANOVA). However, shear bond strength of Fuji IX to dentin was significantly greater than Ketac-Molar (p=0.019) for all groups. Weibull analysis showed that contaminated (Group 2) specimens had a greater probability of failure at low stresses. Modes of failure were mostly cohesive for Fuji IX and adhesive/cohesive for Ketac-Molar. In conclusion, salivary contamination did not affect the mean shear bond strength of Fuji IX GP Fast and Ketac-Molar Maxicap to conditioned dentin; however, it increased the probability of failure at low stresses. PMID- 16382590 TI - Effect of LED curing modes on cusp deflection and hardness of composite restorations. AB - This in vitro study measured cusp deflection associated with MOD resin composite restorations in maxillary premolars with different curing light modes. Soft-start polymerization may reduce cusp deflection by reducing polymerization shrinkage stress. Forty maxillary premolars were mounted in stone and slot MOD cavities were prepared. The teeth were randomized into four groups: Group A--cavities were etched, bonded and restored with two increments of Z-100 composite. Each increment was cured with an LED curing light (fast curing mode). Group B--similar to Group A except that the LED curing light with pulse curing mode was used. Group C--similar to Group A except that the LED curing light with stepped curing mode was used. Group D--a visible curing light was used for curing the composite. The distance between the indexed cusp tips was measured before the restorations were completed and five minutes after, 24 hours after and two weeks after completion of the restorations. The mean contraction of the cusps in microm at five minutes, 24 hours and two weeks, respectively, for each group was A: 25.4, 16.2 and 8.2, B: 6.4, 3.4 and 2.2, C: 11.6, 7.0 and 4.4, D: 33.0, 21.6 and 15.8. Group D resulted in the highest deflection, Group A was intermediate and Groups B and C were the lowest. Ten samples of the composite for each group with 2-mm thickness were prepared for the Vickers hardness test. No difference among the samples was found. PMID- 16382591 TI - Fluoride release/recharge from restorative materials--effect of fluoride gels and time. AB - This study examined the differences in fluoride release and recharge among four restorative materials following treatment with APF or neutral fluoride gel for one or four minutes. Specimens were immersed in 2 mL of deionized water, while fluoride release was measured at 24-hour intervals for 15 days using an ion selective electrode and analyzer. The materials were then treated with the fluoride gels. The fluoride release was measured for 15 days. ANOVA (p<0.05) showed higher fluoride release for Ketac-Fil before fluoride application and for Vitremer and Fuji II LC after application of APF gel. APF gel yielded higher fluoride release when compared to neutral gel, regardless of the material. Fluoride recharge and release was greater after the four-minute APF gel application, with no difference between the times of application for the neutral gel (p>0.05), except for Ketac-Fil. The pattern of release before and after application of the gels was similar and was higher at day 16 compared to day one for the APF gel for resin materials, with higher release at day 15 compared to the initial for Fuji II LC and Vitremer. It was concluded that RM-GICs were the most effective materials with regards to fluoride release after application of APF gel for four minutes. PMID- 16382592 TI - Volumetric polymerization shrinkage of resin composites under simulated intraoral temperature and humidity conditions. AB - This study measured the volumetric shrinkage of resin composites polymerized under temperature and humidity conditions simulating the oral cavity and compared them to those occurring under ambient room conditions. Small, semispherical specimens of a microhybrid (Z100), microfill (Filtek A110) and flowable microhybrid (4 Seasons Flow) resin composite were manually formed and light activated for 40 seconds using a halogen light-curing unit (Spectrum Curing Light). The volumetric polymerization shrinkage of 10 specimens of each brand of resin composite was measured using a drop shape analysis unit (Drop Shape Analysis System, model DSA10 Mk2) under each of two temperature/relative humidity conditions: room conditions (22 +/- 2 degrees C and 60 +/- 5%) and those simulating intraoral conditions (35 degrees C and 92 +/- 5%). Mean volumetric shrinkage values were calculated for each resin composite and the data were analyzed using two-way analysis of variance and t-test (a=0.05) to determine if significant differences existed between the amount of volumetric polymerization shrinkage that occurred under ambient room conditions and that which occurred under simulated intraoral conditions. Mean volumetric shrinkage values measured for the resin composites were: 2.26 +/- 0.04% (ambient) and 2.61 +/- 0.04% (intraoral) for Z100; 1.96 +/- 0.04% (ambient) and 2.28 +/- 0.04% (intraoral) for Filtek A110 and 4.53 +/- 0.06% (ambient) and 5.34 +/- 0.05% (intraoral) for 4 Seasons Flow. For each resin composite, statistical analysis indicated that the amount of volumetric shrinkage measured under simulated intraoral conditions was significantly greater than what was measured under ambient room conditions (p<0.0001). PMID- 16382593 TI - Effects of an experimentally developed adhesive resin system and CO2 laser irradiation on direct pulp capping. AB - This study examined the wound healing process of rat pulp directly capped with various experimentally developed adhesive resin systems and treated with CO2 laser irradiation. The experimental adhesive resins used in this study were made from Clearfil Mega Bond (MB). The adhesive resin groups were capped with a combination of the following primers and bonding agents: commercially available MB primer (MBP), experimental MB primer containing 2wt% N-methacryloyl 5 aminosalicylic acid (5-NMSA: MP3) and 5wt% 12-methacryloyloxydodecylpyridinium bromide (MDPB: ABP); and commercially available MB bonding agent (MBB), experimental MB bonding agent containing 5wt% and 10wt% hydroxyl-calcium phosphate (hydroxyapatite: MB1, MB2) and 5wt% dicalcium phosphate dihydrate (brushite: MB3) as a reparative dentin-promoter. The combination of the three primers and four bonding agents yielded the 12 adhesive resin groups used in this study. The CO2 laser group was irradiated with a laser and directly capped with MB. The CO2 laser used was an Opelaser 03S II SP, and irradiation conditions were as follows: a power output of 0.5 W, superpulse mode 1, repeat pulse mode (a cycle of 10 msec irradiation and 10 msec interval), defocused beam (approximately distance 20 mm from pulp exposure surface) and an irradiation time of three seconds, with air cooling. The control group was capped with Dycal (DY) and MB. After the direct pulp capping procedures were undertaken, all cavities were restored with Clearfil AP-X resin composite. The rats were sacrificed on the 14th post-operative day. The specimens were alternately stained with Mayer's H & E, Hucker-Conn bacterial stain and the ABC method on TGF-beta1. These stained sections were observed using light microscopy and the following parameters were evaluated: pulp tissue disorganization, inflammatory cell infiltration, reparative dentin formation and bacterial penetration. The results of this study include the following: all experimentally developed bonding agent groups showed reparative dentin formation; whereas, the MBB-capped groups showed very little reparative dentin formation. The descending order regarding the amount of reparative dentin formation was MB2 > MB3 > MB1 >>> MBB, which tended to be dependent on the concentration of the blended reparative dentin-promoter. In terms of the quality of the formed dentin, it was observed that MB1-capped teeth tended to form tubular dentin; whereas, MB2- and MB3-capped teeth formed irregular and osteodentin types of dentin. Among the primers used, the descending order regarding the amount of reparative dentin and tubular type dentin formation was MP3 > MBP > ABP. The descending order of migration of macrophages and leukocytes was ABP > MBP > MP3. The CO2 laser group showed a very irregular fibrous dentin matrix in the vicinity of the denatured and carbonized tissue but definite reparative dentin formation was not observed. The control group showed reparative dentin, which was very thick, compared with the other groups. In all the groups, pulp tissue showed almost normal morphology. Positive staining of TGF beta1 was only observed slightly in some specimens of all groups. There was no difference in the staining of each group. Based on the results of this study, it was concluded that the combination of MP3 (containing 2wt% 5-NMSA) and MB1 (containing 5wt% hydroxyapatite) was effective in initiating an early repair process after direct pulp capping. CO2 laser irradiation is effective for field control, but a longer observation time will be required to determine findings concerning dentin bridge formation. PMID- 16382594 TI - Radiographic versus clinical extension of Class II carious lesions using an F speed film. AB - This study investigated the difference in the apparent radiographic and true clinical extension of Class II carious lesions. Sixty-two lesions in both maxillary and mandibular premolars and molars were radiographed using Insight bitewing film. Class II lesions were scored independently by two masked examiners using an 8-point lesion severity scale. During the restoration process the lesions were dissected in a stepwise fashion from the occlusal aspect. Intraoperative photographs (2x) of the lesions were made, utilizing a novel measurement device in the field as a point of reference. Subsequently, the lesions were all given clinical scores using the same 8-point scale. Statistical analysis showed a significant difference between the true clinical extension of the lesions compared to the radiographic score. "Aggressive" and "Conservative" radiographic diagnoses underestimated the true clinical extent by 0.66 mm and 0.91 mm, respectively. No statistical difference was found between premolars and molars or maxillary and mandibular arches. The results of this study help to define the parameters for making restorative treatment decisions involving Class II carious lesions. PMID- 16382595 TI - Occlusal loading evaluation in the cervical integrity of Class II cavities filled with composite. AB - There are many doubts about the clinical behavior of condensable composite restorations in Class II cavities, particularly when they are submitted to axial mechanical loads. This study evaluated cervical microleakage in Class II direct fillings in composite, whether or not they were submitted to an occlusal load cycling. Twenty-three human molars with standardized cavities (proximal vertical "slot") were treated with enamel and cement endings. After completion of the filling process with condensable composite (Surefil), they were separated into two groups: control (without occlusal loading) and test, where 4,000 one-second cycles of 150 N occlusal loading were applied. Twenty teeth were submitted to a microleakage test and then evaluated according to dye penetration. Significant statistical differences (Wilcoxon test, p=0.005<0.05) of leakage degree in enamel and cement were found in the control group. Significant statistical differences at <0.05 were also found in the test group, with p=0.045. After paired comparison of the control and test groups, a significant statistical difference was found at the enamel level (Mann-Whitney test, p=0.03). However, no significant statistical differences were found at the cement level (p=0.28). Therefore, it could be concluded that there was greater microleakage in cement compared to enamel, and occlusal loading has a decisive influence, as it increases the rate of microleakage. PMID- 16382596 TI - Microtensile dentin bond strength of self-etching resins: effect of a hydrophobic layer. AB - In this study, the microtensile bond strength of resin composites to dentin was determined when hydrophilic self-etching resins were used with and without an additional layer of a more hydrophobic adhesive. Included were three single-step self-etching adhesives, Adper Prompt L-Pop (3M ESPE), iBond GI (Heraeus Kulzer, Inc) and Xeno III (Caulk/Dentsply), and as a negative control, UniFil Bond (GC America), a self-etching primer with a separate adhesive. Each product was evaluated using a hybrid resin composite from its respective manufacturer, and each was used as directed and then used with an added layer of a more hydrophobic resin from its respective manufacturer. Testing was performed after 72 hours using a "non-trimming" microtensile test at a crosshead speed of 0.6 mm/minute. When the products were used according to manufacturers' directions, iBond had a significantly higher bond strength to dentin than the other three products (p<0.001), which were not significantly different from each other. For the three self-etching adhesive systems, the addition of a layer of a more hydrophobic resin produced significantly higher bond strengths to dentin (p<0.001), while no significant effect was found for the self-etching primer (p=0.40). A significant interaction was found between the variables product and adhesive treatment. The TEM evaluation of Prompt L-Pop and iBond demonstrated reduced nanoleakage with the additional resin layer. PMID- 16382597 TI - Assessing the tooth-restoration interface wear resistance of two cementation techniques: effect of a surface sealant. AB - This study compared (1) the tooth-restoration interface width of conventional and "resin coating" cementation techniques, (2) the toothbrushing wear resistance of the two interfaces and (3) this study evaluated the influence of a restoration surface sealing on toothbrush wear resistance on both cementation technique interfaces. Mid-coronal buccal surfaces of 40 bovine teeth were ground to obtain a flat enamel surface. For each specimen, a 3 mm x 4 mm x 3 mm dimension rectangular cavity was prepared. The teeth were divided into four groups. Two groups (RC) received a "resin coating" (ED Primer + Tetric Flow) prior to cementation. The remaining two groups (NC) served as non-coated groups. All teeth were restored with composite inlays (Z250) fabricated by the indirect method and were cemented with dual cure resin cement (Panavia F). After finishing the margins, one group from each of the cementation techniques (RC+S and NC+S) had the tooth-restoration interface protected with a restoration surface sealant (Biscover). The specimens were subjected to 100,000 brushing abrasion cycles. The tooth-restoration width was obtained using a Hommel Tester T 1000-basic profilometer and Turbo Datawin NT 1.34 Software (microm). The interface wear (vertical loss/microm and area/microm2) was calculated with Image Tool 3.0 Software. Data were analyzed with Student t-test, one-way analysis of variance and Tukey test (alpha=0.05). Mean interface width for the NC group was 67 microm and 72 microm for the RC group. The student t-test showed no significant differences between groups (p=0.53). ANOVA showed significant differences (p<0.01) in vertical loss among groups (NC: 49.30 microm; NC+S: 7.90 microm; RC: 27.15 microm; RC+S: 4.74 microm). Also, ANOVA showed significant differences (p<0.01) in worn areas among groups (NC: 2,008 microm2; NC+S: 128 microm2; RC: 1,580 microm2 and RC+S: 88 microm2). No differences were found in tooth restoration interface width and worn area between conventional and "resin coating" techniques. "Resin coating" interface presented reduced vertical loss. Restoration surface sealing provided reduced wear in tooth-restoration interface for both techniques. PMID- 16382598 TI - Immediate dentin sealing of onlay preparations: thickness of pre-cured Dentin Bonding Agent and effect of surface cleaning. AB - This study evaluated the thickness of Dentin Bonding Agent (DBA) used for "immediate dentin sealing" of onlay preparations prior to final impression making for indirect restorations. In addition, the amount of DBA that is removed when the adhesive surface is cleaned with polishing or air abrasion prior to final cementation was evaluated. For this purpose, a standardized onlay preparation was prepared in 12 extracted molars, and either OptiBond FL (Kerr) or Syntac Classic (Vivadent) was applied to half of the teeth and cured in the absence of oxygen (air blocking). Each tooth was bisected in a bucco-lingual direction into two sections, and the thickness of the DBA was measured under SEM on gold sputtered epoxy resin replicas at 11 positions. The DBA layer of each half tooth was treated with either air abrasion or polishing. The thickness of the DBAs was then re-measured on the replicas at the same positions. The results were statistically analyzed with non-parametric statistics (Mann-Whitney U test and Kruskal-Wallis test) at a confidence level of 95% (p=0.05). The film thickness of the DBA was not uniform across the adhesive interface (121.13 +/- 107.64 microm), and a great range of values was recorded (0 to 500 microm). Statistically significant differences (p<0.05) were noted, which were both material (OptiBond FL or Syntac Classic) and position (1 to 11) dependent. Syntac Classic presented a higher thickness of DBA (142.34 +/- 125.10 microm) than OptiBond FL (87.99 +/- 73.76 microm). The higher film thickness of both DBAs was at the deepest part of the isthmus (the most concave part of the preparation), while the lowest was at the line angles of the dentinal crest (the most convex part of the preparation). OptiBond FL presented a more uniform thickness around the dentinal crest of preparation; Syntac Classic pooled at the lower parts of the preparation. The amount of DBA that was removed with air abrasion or polishing was not uniform (11.94 +/- 16.46 microm), and a great range of values was recorded (0 to 145 microm). No statistically significant differences (p<0.05) were found either between different DBAs (OptiBond FL or Syntac Classic) or between different treatments (air abrasion or polishing). As far as the effect of different treatments at different positions, polishing removed more DBA from the top of the dentinal crest, but the difference was not statistically significant. Air abrasion removed less DBA from the corners of the dentinal crest (Positions 4 and 6) than the outer buccal part of the preparation (Positions 1 and 2). Neither air abrasion nor polishing removed the entire layer thickness of the DBA in the majority of the cases. PMID- 16382599 TI - Curing efficacy of a new generation high-power LED lamp. AB - This study investigated the curing efficacy of a new generation high-power LED lamp (Elipar Freelight 2 [N] 3M-ESPE). The effectiveness of composite cure with this new lamp was compared to conventional LED/halogen (Elipar Freelight [F], 3M ESPE; Max [M], Dentsply-Caulk) and high-power halogen (Elipar Trilight [T], 3M ESPE; Astralis 10 [A], Ivoclar Vivadent) lamps. Standard continuous (NS, FS, TS; MS), turbo (AT) and exponential (NE, FE, TE) curing modes of the various lights were examined. Curing efficacy of the various lights and modes were determined by measuring the top and bottom surface hardness of 2-mm thick composite specimens (Z100, 3M-ESPE) using a digital microhardness tester (n=5; load=500 g; dwell time=15 seconds) one hour after light polymerization. The hardness ratio was computed by dividing HK (Knoops Hardness) of the bottom surface by HK of the top surface. The data was analyzed using one-way ANOVA/Scheffe's test and Independent Samples t-test at significance level 0.05. Results of the statistical analysis were as follows: HK top--E, FE, NE > NS and NE > AT, TS, FS; HK bottom--TE, NE > NS; Hardness ratio--NS > FE and FS, TS > NE. No significant difference in HK bottom and hardness ratio was observed between the two modes of Freelight 2 and Max. Freelight 2 cured composites as effectively as conventional LED/halogen and high-power halogen lamps, even with a 50% reduction in cure time. The exponential modes of Freelight 2, Freelight and Trilight appear to be more effective than their respective standard modes. PMID- 16382600 TI - Curing of pit & fissure sealants using Light Emitting Diode curing units. AB - Light Emitting Diode (LED) curing units are attractive to clinicians, because most are cordless and should create less heat within tooth structure. However, questions about polymerization efficacy have surrounded this technology. This research evaluated the adequacy of the depth of cure of pit & fissure sealants provided by LED curing units. Optilux (OP) and Elipar Highlight (HL) high intensity halogen and Astralis 5 (A5) conventional halogen lights were used for comparison. The Light Emitting Diode (LED) curing units were Allegro (AL), LE Demetron I (DM), FreeLight (FL), UltraLume 2(UL), UltraLume 5 (UL5) and VersaLux (VX). Sealants used in the study were UltraSeal XT plus Clear (Uclr), Opaque (Uopq) and Teethmate F-1 Natural (Kclr) and Opaque (Kopq). Specimens were fabricated in a brass mold (2 mm thick x 6 mm diameter) and placed between two glass slides (n=5). Each specimen was cured from the top surface only. One hour after curing, four Knoop Hardness readings were made for each top and bottom surface at least 1 mm from the edge. The bottom to top (B/T) KHN ratio was calculated. Groups were fabricated with 20 and 40-second exposure times. In addition, a group using a 1 mm-thick mold was fabricated using an exposure time of 20 seconds. Differences between lights for each material at each testing condition were determined using one-way ANOVA and Student-Newman-Keuls Post-hoc test (alpha=0.05). There was no statistical difference between light curing units for Uclr cured in a 1-mm thickness for 20 seconds or cured in a 2 mm-thickness for 40 seconds. All other materials and conditions showed differences between light curing units. Both opaque materials showed significant variations in B/T KHN ratios dependent upon the light-curing unit. PMID- 16382601 TI - American Academy of Gold Foil Operators Clinician of the Year Award. Dr Barry O Evans. PMID- 16382602 TI - American Academy of Gold Foil Operators Distinguished Member Award. Dr Michael A Cochran. PMID- 16382603 TI - Visualization task performance with 2D, 3D, and combination displays. AB - We describe a series of experiments that compare 2D displays, 3D displays, and combined 2D/3D displays (orientation icon, ExoVis, and clip planes) for relative position estimation, orientation, and volume of interest tasks. Our results indicate that 3D displays can be very effective for approximate navigation and relative positioning when appropriate cues, such as shadows, are present. However, 3D displays are not effective for precise navigation and positioning except possibly in specific circumstances, for instance, when good viewing angles or measurement tools are available. For precise tasks in other situations, orientation icon and ExoVis displays were better than strict 2D or 3D displays (displays consisting exclusively of 2D or 3D views). The combined displays had as good or better performance, inspired higher confidence, and allowed natural, integrated navigation. Clip plane displays were not effective for 3D orientation because users could not easily view more than one 2D slice at a time and had to frequently change the visibility of individual slices. Major factors contributing to display preference and usability were task characteristics, orientation cues, occlusion, and spatial proximity of views that were used together. PMID- 16382604 TI - Time-varying contour topology. AB - The contour tree has been used to compute the topology of isosurfaces, generate a minimal seed set for accelerated isosurface extraction, and provide a user interface to segment individual contour components in a scalar field. In this paper, we extend the benefits of the contour tree to time-varying data visualization. We define temporal correspondence of contour components and describe an algorithm to compute the correspondence information in time-dependent contour trees. A graph representing the topology changes of time-varying isosurfaces is constructed in real-time for any selected isovalue using the precomputed correspondence information. Quantitative properties, such as surface area and volume of contour components, are computed and labeled on the graph. This topology change graph helps users to detect significant topological and geometric changes in time-varying isosurfaces. The graph is also used as an interactive user interface to segment, track, and visualize the evolution of any selected contour components over time. PMID- 16382605 TI - Distance preserving flattening of surface sections. AB - Curved cross-sections extracted from medical volume images are useful for analyzing nonplanar anatomic structures such as the aorta arch or the pelvis. For visualization and for performing distance measurements, extracted surface sections need to be adequately flattened. We present two different distance preserving surface flattening methods which preserve distances according to a user-specified center of interest and according to user-specified orientations. The first method flattens surface sections by preserving distances along surface curves located within planes having a user specified constant orientation. The second method flattens surfaces along curves located within radial planes crossing the center of interest. We study and compare the properties of the two flattening methods by analyzing their distortion maps. Thanks to a multiresolution approach, we provide surface flattening at interactive rates, allowing users to displace their focus point while visualizing the resulting flattened surface. These distance preserving flattening methods provide new means of inspecting curved cross-sections extracted from medical images. PMID- 16382606 TI - Realistic haptic rendering of interacting deformable objects in virtual environments. AB - A new computer haptics algorithm to be used in general interactive manipulations of deformable virtual objects is presented. In multimodal interactive simulations, haptic feedback computation often comes from contact forces. Subsequently, the fidelity of haptic rendering depends significantly on contact space modeling. Contact and friction laws between deformable models are often simplified in up to date methods. They do not allow a "realistic" rendering of the subtleties of contact space physical phenomena (such as slip and stick effects due to friction or mechanical coupling between contacts). In this paper, we use Signorini's contact law and Coulomb's friction law as a computer haptics basis. Real-time performance is made possible thanks to a linearization of the behavior in the contact space, formulated as the so-called Delassus operator, and iteratively solved by a Gauss-Seidel type algorithm. Dynamic deformation uses corotational global formulation to obtain the Delassus operator in which the mass and stiffness ratio are dissociated from the simulation time step. This last point is crucial to keep stable haptic feedback. This global approach has been packaged, implemented, and tested. Stable and realistic 6D haptic feedback is demonstrated through a clipping task experiment. PMID- 16382607 TI - Geometry-driven photorealistic facial expression synthesis. AB - Expression mapping (also called performance driven animation) has been a popular method for generating facial animations. A shortcoming of this method is that it does not generate expression details such as the wrinkles due to skin deformations. In this paper, we provide a solution to this problem. We have developed a geometry-driven facial expression synthesis system. Given feature point positions (the geometry) of a facial expression, our system automatically synthesizes a corresponding expression image that includes photorealistic and natural looking expression details. Due to the difficulty of point tracking, the number of feature points required by the synthesis system is, in general, more than what is directly available from a performance sequence. We have developed a technique to infer the missing feature point motions from the tracked subset by using an example-based approach. Another application of our system is expression editing where the user drags feature points while the system interactively generates facial expressions with skin deformation details. PMID- 16382608 TI - Stylized and abstract painterly rendering system using a multiscale segmented sphere hierarchy. AB - This paper presents a novel system framework for interactive, three-dimensional, stylized, abstract painterly rendering. In this framework, the input models are first represented using 3D point sets and then this point-based representation is used to build a multiresolution bounding sphere hierarchy. From the leaf to root nodes, spheres of various sizes are rendered into multiple-size strokes on the canvas. The proposed sphere hierarchy is developed using multiscale region segmentation. This segmentation task assembles spheres with similar attribute regularities into a meaningful region hierarchy. These attributes include colors, positions, and curvatures. This hierarchy is very useful in the following respects: 1) it ensures the screen-space stroke density, 2) controls different input model abstractions, 3) maintains region structures such as the edges/boundaries at different scales, and 4) renders models interactively. By choosing suitable abstractions, brush stroke, and lighting parameters, we can interactively generate various painterly styles. We also propose a novel scheme that reduces the popping effect in animation sequences. Many different stylized images can be generated using the proposed framework. PMID- 16382609 TI - A new combinatorial approach to surface reconstruction with sharp features. AB - This paper presents a new combinatorial approach to surface reconstruction with sharp features. Different from other postprocessing methods, the proposed method provides a systematic way to identify and reconstruct sharp features from unorganized sample points in one integrated reconstruction process. In addition, unlike other approximation methods, the reconstructed triangulated surface is guaranteed to pass through the original sample points. In this paper, the sample points in the sharp regions are defined as characteristic vertices (c-vertices), and their associated poles (c-poles) are used as a "sculptor" to extract triangles from a Delaunay structure for the sharp features. But, for smooth surface regions, an efficient region-growing scheme is used for triangle extraction and connection. Since only the c-poles associated with the sharp regions are used to participate in the Delaunay computation with the sample points, the proposed algorithm is adaptive in the sense that, given a sampled object with less sharp features, the triangulation becomes more efficient. To validate the proposed algorithm, some detailed illustrations are given. Experimental results show that it is robust and highly efficient. PMID- 16382610 TI - Mesh parameterization by minimizing the synthesized distortion metric with the coefficient-optimizing algorithm. AB - The parameterization of a 3D mesh into a planar domain requires a distortion metric and a minimizing process. Most previous work has sought to minimize the average area distortion, the average angle distortion, or a combination of these. Typical distortion metrics can reflect the overall performance of parameterizations but discount high local deformations. This affects the performance of postprocessing operations such as uniform remeshing and texture mapping. This paper introduces a new metric that synthesizes the average distortions and the variances of both the area deformations and the angle deformations over an entire mesh. Experiments show that, when compared with previous work, the use of synthesized distortion metric performs satisfactorily in terms of both the average area deformation and the average angle deformation; furthermore, the area and angle deformations are distributed more uniformly. This paper also develops a new iterative process for minimizing the synthesized distortion, the coefficient-optimizing algorithm. At each iteration, rather than updating the positions immediately after the local optimization, the coefficient optimizing algorithm first update the coefficients for the linear convex combination and then globally updates the positions by solving the Laplace system. The high performance of the coefficient-optimizing algorithm has been demonstrated in many experiments. PMID- 16382611 TI - Thin structure segmentation and visualization in three-dimensional biomedical images: a shape-based approach. AB - This paper presents a shape-based approach in extracting thin structures, such as lines and sheets, from three-dimensional (3D) biomedical images. Of particular interest is the capability to recover cellular structures, such as microtubule spindle fibers and plasma membranes, from laser scanning confocal microscopic (LSCM) data. Hessian-based shape methods are reviewed. A synthesized linear structure is used to evaluate the sensitivity of the multiscale filtering approach in extracting closely positioned fibers. We find that the multiscale approach tends to fuse lines together, which makes it unsuitable for visualizing mouse egg spindle fibers. Single-scale Gaussian filters, balanced between sensitivity and noise resistance, are adopted instead. In addition, through an ellipsoidal Gaussian model, the eigenvalues of the Hessian matrix are quantitatively associated with the standard deviations of the Gaussian model. Existing shape filters are simplified and applied to LSCM data. A significant improvement in extracting closely positioned thin lines is demonstrated by the resultant images. Further, the direct association of shape models and eigenvalues makes the processed images more understandable qualitatively and quantitatively. PMID- 16382612 TI - Predictive haptic guidance: intelligent user assistance for the control of dynamic tasks. AB - Intelligent systems are increasingly able to offer real-time information relevant to a user's manual control of an interactive system, such as dynamic system control space constraints for animation control and driving. However, it is difficult to present this information in a usable manner and other approaches which have employed haptic cues for manual control in "slow" systems often lead to instabilities in highly dynamic tasks. We present a predictive haptic guidance method based on a look-ahead algorithm, along with a user evaluation which compares it with other approaches (no guidance and a standard potential-field method) in a 1-DoF steered path-following scenario. Look-ahead guidance outperformed the other methods in both quantitative performance and subjective preference across a range of path complexity and visibility and a force analysis demonstrated that it applied smaller and fewer forces to users. These results (which appear to derive from the predictive guidance's supporting users in taking earlier and more subtle corrective action) suggest the potential of predictive methods in aiding manual control of dynamic interactive tasks where intelligent support is available. PMID- 16382613 TI - Ray-tracing polymorphic multidomain spectral/hp elements for isosurface rendering. AB - The purpose of this paper is to present a ray-tracing isosurface rendering algorithm for spectral/hp (high-order finite) element methods in which the visualization error is both quantified and minimized. Determination of the ray isosurface intersection is accomplished by classic polynomial root-finding applied to a polynomial approximation obtained by projecting the finite element solution over element-partitioned segments along the ray. Combining the smoothness properties of spectral/hp elements with classic orthogonal polynomial approximation theory, we devise an adaptive scheme which allows the polynomial approximation along a ray-segment to be arbitrarily close to the true solution. The resulting images converge toward a pixel-exact image at a rate far faster than sampling the spectral/hp element solution and applying classic low-order visualization techniques such as marching cubes. PMID- 16382614 TI - Frequency-temperature compensation of piezoelectric resonators by electric DC bias field. AB - Electromechanical resonators have been widely used in signal processing and frequency control applications. It has been found that the resonant frequency of most resonator devices is highly temperature dependent, as temperature variation leads to materials properties change as well as resonator dimension change, which result in the undesirable shift of the resonance frequency. In this paper, we present a new frequency tuning method in which direct current (DC) bias field is used to control the resonance frequency of the piezoelectric resonator that is subjected to ambient temperature variations. It has been found that, depending on the polarity, the application of a DC bias field can reduce or increase the resonance frequency of the resonator. The experimental results demonstrate that the DC bias field tuning can achieve fairly good temperature compensation within a certain temperature range, and that the mechanical Q factor of the resonator is quite stable under different DC bias fields. PMID- 16382615 TI - Electromechanical properties of high coupling single crystals under large electric drive and uniaxial compression. AB - This work investigates the 33-mode electromechanical response of relaxor ferroelectric lead magnesium niobate-lead titanate (PMN-PT) single crystals when driven with large fields approximately 0.4 MV/m under a combined direct current (DC) field and mechanical bias similar to those used in the design of sound projectors. It demonstrates that the remarkable small signal length extensional coupling (k33 > 0.90) and other electromechanical properties of morphotropic PMN PT single crystals prevail under large drive. The observed k33 roll-off at 42 MPa compressive stress is analyzed in terms of the recent structural data and the high-order Devonshire theory of possible ferroelectric-ferroelectric transition trajectories. PMID- 16382616 TI - Designs of a microwave TE011 mode cavity for a space borne H-maser. AB - Method of Lines and Finite Element Analysis investigations have been performed to optimize parameters in a TE011 mode cavity resonator suitable for a spaceborne hydrogen maser. We report on designs that were explored to find a global maximum in the important design parameters for the microwave cavity used in a hydrogen maser. The criteria sought in this exercise were both the minimization of the total volume of the cavity and the maximization of the product of the z-component of the magnetic energy filling factor and the cavity TE011 mode Q-factor (Q.eta). Different configurations were studied. They were a sapphire tube in a copper cylinder, a sapphire tube in a copper cylinder with Bragg reflectors, and spherical copper cavities both empty and sapphire-lined on the inside cavity surface. At 320 K, the simulations resulted in an optimum product Q.eta = 4.9 x 10(4), with an inner cavity radius of 80 mm and unity aspect ratio. This represents a 54% improvement over an earlier design. The expected increase in the product Q . eta) with the inclusion of Bragg reflectors to the sapphire tube was not achieved. Moreover, the z-component of the magnetic energy filling factor was greatly reduced due to an increase in the radial magnetic field. The sapphire lined spherical cavity showed no better performance than an equivalent-sized empty copper spherical cavity. For the empty cavity the simulations resulted in the product Q.eta = 4.4 x 10(4). The empty spherical cavity resonator is not suitable for the spaceborne hydrogen maser as the total volume in this case is 33% larger than that of the optimized sapphire tube resonator. PMID- 16382617 TI - Temperature dependence of ultrasonic backscattered energy in motion-compensated images. AB - Noninvasive temperature imaging would enhance the ability to uniformly heat tumors at therapeutic levels. Ultrasound is an attractive modality for this purpose. Previously, we predicted monotonic changes in backscattered energy (CBE) of ultrasound with temperature for certain subwavelength scatterers. We also measured CBE values similar to our predictions in bovine liver, turkey breast, and pork muscle in one dimension (1-D). Those measurements were corrected manually for changes in the axial position of echo signals with temperature. To investigate the effect of temperature on CBE in 2-D, we imaged 1-cm thick samples of bovine liver, turkey breast, and pork muscle during heating in a water bath. Images were formed by a phased-array imager with a 7 MHz linear probe. Using radio frequency (RF) signals permitted the use of cross correlation as a similarity measure for automatic tracking of feature displacement as a function of temperature. Feature displacement across the specimen was nonuniform with typical total displacements of 0.5 mm in both axial and lateral directions. Apparent movement in eight image regions in each specimen was tracked from 37 to 50 degrees C in 0.5 degrees C steps. Envelopes of motion-compensated image regions were found then smoothed with a 3 x 3 running average filter before forming the backscattered energy at each pixel. Our measure of CBE compared means of both the positive and negative changes in the backscattered energy (BE) images. CBE was monotonic and differed by about 4 dB at 50 degrees C from its value at 37 degrees C. Relatively noise-free CBE curves from tissue volumes of less than 1 cm3 supports the use of CBE for temperature estimation. PMID- 16382618 TI - Comparative evaluation of despeckle filtering in ultrasound imaging of the carotid artery. AB - It is well-known that speckle is a multiplicative noise that degrades the visual evaluation in ultrasound imaging. The recent advancements in ultrasound instrumentation and portable ultrasound devices necessitate the need of more robust despeckling techniques for enhanced ultrasound medical imaging for both routine clinical practice and teleconsultation. The objective of this work was to carry out a comparative evaluation of despeckle filtering based on texture analysis, image quality evaluation metrics, and visual evaluation by medical experts in the assessment of 440 (220 asymptomatic and 220 symptomatic) ultrasound images of the carotid artery bifurcation. In this paper a total of 10 despeckle filters were evaluated based on local statistics, median filtering, pixel homogeneity, geometric filtering, homomorphic filtering, anisotropic diffusion, nonlinear coherence diffusion, and wavelet filtering. The results of this study suggest that the first order statistics filter lsmv, gave the best performance, followed by the geometric filter gf4d, and the homogeneous mask area filter lsminsc. These filters improved the class separation between the asymptomatic and the symptomatic classes based on the statistics of the extracted texture features, gave only a marginal improvement in the classification success rate, and improved the visual assessment carried out by the two experts. More specifically, filters lsmv or gf4d can be used for despeckling asymptomatic images in which the expert is interested mainly in the plaque composition and texture analysis; and filters lsmv, gf4d, or lsminsc can be used for the despeckling of symptomatic images in which the expert is interested in identifying the degree of stenosis and the plaque borders. The proper selection of a despeckle filter is very important in the enhancement of ultrasonic imaging of the carotid artery. Further work is needed to evaluate at a larger scale and in clinical practice the performance of the proposed despeckle filters in the automated segmentation, texture analysis, and classification of carotid ultrasound imaging. PMID- 16382619 TI - Usefulness of ultrasonic strain measurement-based shear modulus reconstruction for diagnosis and thermal treatment. AB - We previously reported an ultrasonic strain measurement-based one-dimensional (1 D) shear modulus reconstruction technique using a regularization method for differential diagnosis of malignancies on human superficial tissues (e.g., breast tissues). Here, ultrasonic strain measurement-based 2-D and 3-D shear modulus reconstruction techniques are described, and the 1-D technique is reviewed and subsequently applied to various human in vivo tissues, including deeply situated tissues (e.g., liver). Because soft tissues are deformed in 3-D space by externally situated arbitrary mechanical sources, the accuracy of the low dimensional (i.e., 1-D or 2-D) reconstructions is lower to that of 3-D reconstruction due to occurrence of erroneous reconstruction artifacts (i.e., the reconstructed modulus is different than reality). These artifacts are confirmed on simulated inhomogeneous cubic phantoms containing a spherical homogenous inclusion using numerically calculated deformation data. The superiority of quasi real-time imaging of the shear modulus is then demonstrated by comparing it with conventional B-mode imaging and strain imaging from the standpoints of monitoring the effectiveness of minimally invasive thermal therapy as well as differential diagnosis. Because the 2-D and 3-D techniques require special ultrasonic (US) equipment, the 1-D technique using conventional US imaging equipment is used, even though erroneous artifacts will occur. Specifically, the 1-D technique is applied as a diagnostic tool for differentiating malignancies in human in vivo liver and breast tissue, and a monitoring technique for determining the effectiveness of interstitial electromagnetic wave (micro and rf) thermal therapy on human in vivo liver and calf in vitro liver. Even when using the 1-D technique, reconstructed shear moduli were confirmed to be a suitable measure for monitoring thermal treatment as well as differential diagnosis. These results are encouraging in that they will promote use of 2-D and 3-D reconstruction techniques. PMID- 16382620 TI - In vivo acceleration of ultrasonic tissue heating by microbubble agent. AB - The ultrasonic power absorbed by a microbubble in its continuous wave response is estimated through numerically solving a version of the Rayleigh-Plesset equation. At an ultrasonic frequency of 3 MHz, a resonant microbubble, approximately 1.1 microm in radius, showed an absorption cross section of about 0.005 mm2 in its low power response. This estimation predicts that the tissue ultrasonic absorption will be doubled when such microbubbles are delivered to the tissue at a concentration of about eight bubbles/mm3 in tissue. An exteriorized murine kidney was exposed to focused ultrasound at 3.2 MHz in degassed saline, and the tissue temperature change was measured. With an intravenous bolus administration of a microbubble agent, the ultrasonically induced temperature elevation was multiplied by up to five times. The enhancement in temperature elevation gradually decreased as the microbubble agent was eliminated from the body. The experimental results agreed with the prediction in the order of magnitude. This effect may have a potential use to enhance the throughput as well as the selectivity of focused ultrasound treatment. PMID- 16382621 TI - A finite-element method model of soft tissue response to impulsive acoustic radiation force. AB - Several groups are studying acoustic radiation force and its ability to image the mechanical properties of tissue. Acoustic radiation force impulse (ARFI) imaging is one modality using standard diagnostic ultrasound scanners to generate localized, impulsive, acoustic radiation forces in tissue. The dynamic response of tissue is measured via conventional ultrasonic speckle-tracking methods and provides information about the mechanical properties of tissue. A finite-element method (FEM) model has been developed that simulates the dynamic response of tissues, with and without spherical inclusions, to an impulsive acoustic radiation force excitation from a linear array transducer. These FEM models were validated with calibrated phantoms. Shear wave speed, and therefore elasticity, dictates tissue relaxation following ARFI excitation, but Poisson's ratio and density do not significantly alter tissue relaxation rates. Increased acoustic attenuation in tissue increases the relative amount of tissue displacement in the near field compared with the focal depth, but relaxation rates are not altered. Applications of this model include improving image quality, and distilling material and structural information from tissue's dynamic response to ARFI excitation. Future work on these models includes incorporation of viscous material properties and modeling the ultrasonic tracking of displaced scatterers. PMID- 16382622 TI - Finite-element analysis of temperature rise and lesion formation from catheter ultrasound ablation transducers. AB - A model using finite-element analysis (FEA) has been developed to calculate the temperature rise in tissue from intracardiac ultrasound ablation catheters and to predict if this temperature rise is adequate for producing a lesion in the tissue. In the model, acoustic fields are simulated with Field II, and heat transfer is modeled with an FEA program. To validate the model, we compare its results to experimental results from an integrated, real-time three-dimensional (3-D) ultrasound imaging and ultrasound ablation catheter. The ultrasound ablation transducer is a ring transmitting at 10 MHz capable of producing an acoustic intensity of 16 W/cm2. It was used to ablate four lesions in tissue, and temperature rise as a function of time was monitored by embedded thermocouples. The average absolute difference between final temperatures predicted by FEA and those measured is 1.95 +/- 0.72 degrees C. Additionally, model and experimental lesion size are in good agreement. The model then is used to design a new ultrasound catheter with a 7.5 MHz linear phased array for ablation. Eight designs are modeled, and acoustic intensity, temperature rise, and ablation ability are compared. PMID- 16382623 TI - Estimation of the squeeze film effect in a surface acoustic wave motor. AB - This paper describes the air film characteristic in the interface between the slider and stator substrate of a surface acoustic wave (SAW) motor, and the difference of the squeeze film effect by the form of the slider. Because the vibration amplitude of the SAW motor is several 10 nm order, analysis of pressure was performed by using the molecular gas-film lubrication (MGL) equation. In the analysis, the MGL equation was split into advection phase and nonadvection phase, and calculation of advection and nonadvection phases was performed by using the cubic interpolated propagation (CIP) method and the finite difference method, respectively. From analysis results it was found that the time to steady pressure of air film depends on the radius of projection arranged on the contact surface of the slider. Also found was that the steady pressure in the interface does not depend on the radius and height of the projection, but it depends on the minimum spacing between the slider and the stator substrate. PMID- 16382624 TI - An ultrasonic linear motor using ridge-mode traveling waves. AB - A new type of ultrasonic linear motor is presented using traveling waves excited along a ridge atop a substrate. The ridge cross section was designed to permit only the fundamental mode to be excited during operation of the motor, with a Langevin transducer used as the source of vibration in this study. The ridge waveguide was first made of lossy media to avoid reflecting vibration energy back toward the vibration source, forming a traveling wave. A 5-mm-wide, 15-mm-tall rectangular acrylic ridge was used to move a slider placed upon it toward the vibration source, in opposition to the direction of the traveling wave transmitted along the waveguide ridge. Using a low-loss 3 x 6-mm aluminum rectangular ridge combined with a damper clamped onto the far end of the waveguide, similar results were obtained. To obtain bidirectional operation, the damper was replaced with a second Langevin transducer, giving a pair of transducers located perpendicularly to the ends of the ridge and driven with an appropriate phase difference. The moving direction of the slider was reversed by shifting this phase difference by about 180 degrees. With this simple configuration, it may soon be possible to fabricate a linear micromotor system on a silicon substrate or other semiconductor wafer adjacent to other electronic and optoelectronic devices. PMID- 16382625 TI - Ultrasonic wave transmission in periodically undulated plates. AB - This study focuses on quantifying the change in phase speed of waves transmitting through periodically undulated plates under pass band interaction. A perturbation technique is used to analyze the transmission of horizontally polarized guided waves in elastic plates with sinusoidal periodicity at their outerfaces. Phase speed of transmitting modes is presented as a function of various parameters, including outerface wavenumber, undulation amplitude, degree of undulations symmetry about the periodically undulated plate midplane, plate average thickness, and frequency of propagation. PMID- 16382626 TI - Bending mode effect on sensitivity of plate surface acoustic wave pressure sensors. AB - The bending mode effect on sensitivity of pressure sensors operating with surface acoustic waves (SAW) propagating over the surface of a plate is analyzed using Tiersten's perturbation integral for frequency shifts in a piezoelectric resonator due to the presence of initial fields. For a plate subjected to nonpure bending, the SAW speed, whose shift in response to pressure determines the sensitivity, can be defined locally through the local variations of the effective material constants. Three common bending modes of pure bending, bending under a concentrate load, and bending under a uniformly load are analyzed and compared. PMID- 16382627 TI - Errors and uncertainties in the measurement of ultrasonic wave attenuation and phase velocity. AB - This paper presents an analysis of the error generation mechanisms that affect the accuracy of measurements of ultrasonic wave attenuation coefficient and phase velocity as functions of frequency. In the first stage of the analysis we show that electronic system noise, expressed in the frequency domain, maps into errors in the attenuation and the phase velocity spectra in a highly nonlinear way; the condition for minimum error is when the total measured attenuation is around 1 Neper. The maximum measurable total attenuation has a practical limit of around 6 Nepers and the minimum measurable value is around 0.1 Neper. In the second part of the paper we consider electronic noise as the primary source of measurement error; errors in attenuation result from additive noise whereas errors in phase velocity result from both additive noise and system timing jitter. Quantization noise can be neglected if the amplitude of the additive noise is comparable with the quantization step, and coherent averaging is employed. Experimental results are presented which confirm the relationship between electronic noise and measurement errors. The analytical technique is applicable to the design of ultrasonic spectrometers, formal assessment of the accuracy of ultrasonic measurements, and the optimization of signal processing procedures to achieve a specified accuracy. PMID- 16382628 TI - Detection of structural damage from the local temporal coherence of diffuse ultrasonic signals. AB - Permanently mounted ultrasonic transducers have the potential to interrogate large areas of a structure, and thus be effective global sensors for structural health monitoring. Recorded signals, although very sensitive to damage, are long, complex, and difficult to interpret compared to pulse echo and through transmission signals customary for nondestructive testing. These diffuse signals also are quite sensitive to environmental effects such as temperature and surface condition changes. Waveform comparison methods such as time domain differencing and spectral analysis, although effective for detecting changes, are generally unsuccessful in discriminating damage from environmental effects. This paper considers the local temporal coherence as another means of comparing two waveforms in order to provide a quantitative measure of the change in shape of a signal compared to a reference as a function of time from transmit. Experimental results show that the local temporal coherence is effective in discriminating structural damage from both temperature changes and modest changes in surface conditions; results are compared to those obtained from time domain and spectrogram differencing. The advantages of this methodology are the simplicity of the transducers, the applicability to a wide range of structures, and the straightforward signal processing. PMID- 16382629 TI - A one-dimensional numerical model of acoustic wave propagation in a multilayered structure of a resistance spot weld. AB - A one-dimensional model of acoustic wave propagation in a multilayered structure of a spot weld is developed. The inhomogeneity of the material properties due to the thermal inhomogeneity is included in the equation of motion. The model enables us to deal with arbitrary spatial distributions of Lame constants and density. The model allows analysis of travel time, multiple reflections, and interference in a given geometry. Use of this model could provide information to help predict behavior of the waves in the transmission (reflection) mode at different plate thicknesses and welding settings. PMID- 16382630 TI - Apparent texture symmetry deviations in aluminum sheet. AB - An ultrasonic texture measurement system for sheet metal is being developed using rotating electromagnetic acoustic transducers (EMATs). We report on further investigations of deviation from theoretically predicted symmetries in the elastic constants (as measured ultrasonically using the aforementioned system) of cold-rolled aluminum sheet reported in earlier publications. A study of the effects of annealing and deliberate deformation (both elastic and plastic) are used to develop an explanation of the nature and likely origin of this asymmetry. These deviations from symmetry cannot be detected by ultrasonic Lamb wave measurement in three directions alone. Texture asymmetry is relevant to the sheet metal manufacturer as it affects formability and may indicate processing problems. Results indicate that the asymmetry in the measured ultrasonic Lamb wave velocities on either side of the rolling direction is due to a stress effect rather than crystallographic PMID- 16382631 TI - Approximate estimates of limiting errors of passive wireless SAW sensing with DPM. AB - This paper discusses approximate statistical estimates of limiting errors associated with single differential phase measurement of a time delay (phase difference) between two reflectors of the passive surface acoustic wave (SAW) sensor. The remote wireless measurement is provided at the ideal coherent receiver using the maximum likelihood function approach. Approximate estimates of the mean error, mean square error, estimate variance, and Cramer-Rao bound are derived along with the error probability to exceed a threshold in a wide range of signal-to-noise ratio (SNR) values. The von Mises/Tikhonov distribution is used as an approximation for the phase difference and differential phase diversity. Simulation of the random phase difference and limiting errors also is applied. PMID- 16382632 TI - Growth of A1N piezoelectric film on diamond for high-frequency surface acoustic wave devices. AB - Diamond films are very desirable for application to SAW devices because of their high acoustic wave velocity, which allows the extending of the frequency limit of operation at a given interdigital transducer line-width resolution. Use of high quality AIN as the piezoelectric layer in conjunction with diamond is also desirable because of its high SAW velocity--the highest among all piezoelectric materials--together with its excellent electrical, mechanical, and chemical properties. The problems arising in the growth of A1N films on diamond have prevented, until now, the use of this combination of materials. In this paper we present recent results on the growth of highly oriented, low-stressed A1N films on diamond. SAW propagation on A1N/diamond has been theoretically investigated together with electromechanical coupling for both the Rayleigh and the Sezawa modes. The theoretical calculations show that high SAW velocities are achievable with good coupling efficiencies. Under proper conditions very large piezoelectric couplings are predicted--k2 = 2.2 and 4% for the Rayleigh and the Sezawa wave, respectively--comparable to those observed in strongly piezoelectric single crystals such as LiNbO3, but with SAW velocities approximately two-fold higher. Experiments performed on A1N/diamond/Si SAW test devices have shown good agreement between experimental results and theoretical predictions and demonstrate the feasibility of SAW devices based on this technology. PMID- 16382633 TI - Q values of longitudinal leaky Saws propagating on rotated Y-cut LN substrates along the perpendicular to the X axis. AB - Longitudinal leaky SAWs (LLSAWs), which can be excited in a thick aluminum (A1) grating on a rotated Y-cut lithium-niobate substrate, were analyzed in a simulation that combined both FEM and BEM. The Q-value dependence of an LLSAW on the rotated angle and the Al-grating thickness was clear. The simulation results were confirmed experimentally. These SAWs have potential applications in high frequency SAW devices because their velocities range from 6100 to 6500 m/s and their optimum Q values are theoretically larger than 4000. PMID- 16382634 TI - Effect of crystalline quality of diamond film to the propagation loss of surface acoustic wave devices. AB - Diamond films with various crystal qualities were grown by chemical vapor deposition on silicon wafers. Their crystallinity was characterized by Raman scattering and electron backscattering diffraction. By fabricating a device structure for surface acoustic wave (SAW) using these diamond films, the propagation loss was measured at 1.8 GHz and compared with the crystallinity. It was found that the propagation loss was lowered in relatively degraded films having small crystallites, a narrow distribution in the diamond crystallite size, and preferential grain orientation. This experiment clarifies diamond film characteristics required for high-frequency applications in SAW filters. PMID- 16382635 TI - Piezoelectric ceramics with high dielectric constants for ultrasonic medical transducers. AB - Complex system ceramics Pb(Sc(1/2)Nb(1/2))O3-Pb(Mg(1/3)Nb(2/3))O3 Pb(Ni(1/2)Nb(1/2))O3-(Pb0.965,Sr0.035) (Zr,Ti)O3 (PSN-PMN-PNN-PSZT abbreviated PSMNZT) have been synthesized by the conventional technique, and dielectric and piezoelectric properties of the ceramics have been investigated for ultrasonic medical transducers. High capacitances of the transducers are desired in order to match the electrical impedance between the transducers and the coaxial cable in array probes. Although piezoelectric ceramics that have high dielectric constants (epsilon33t/epsilon0 > 5000, k'33 < 70%) are produced in many foundries, the dielectric constants are insufficient. However, we have reported that low molecular mass B-site ions in the lead-perovskite structures are important in realizing better dielectric and piezoelectric properties. We focused on the complex system ceramics PSMNZT that consists of light B-site elements. The maximum dielectric constant, epsilon33T/epsilon0 = 7, 200, was confirmed in the ceramics, where k'33 = 69%, d33 = 940 pC/N, and T(c) = 135 degrees C were obtained. Moreover, pulse-echo characteristics were simulated using the Mason model. The PSMNZT ceramic probe showed echo amplitude about 5.5 dB higher than that of the conventional PZT ceramic probe (PZT-5H type). In this paper, the electrical properties of the PSMNZT ceramics and the simulation results for pulse echo characteristics of the phased-array probes are introduced. PMID- 16382636 TI - Electrical and mechanical fully coupled theory and experimental verification of Rosen-type piezoelectric transformers. AB - A piezoelectric transformer is a power transfer device that converts its input and output voltage as well as current by effectively using electrical and mechanical coupling effects of piezoelectric materials. Equivalent-circuit models, which are traditionally used to analyze piezoelectric transformers, merge each mechanical resonance effect into a series of ordinary differential equations. Because of using ordinary differential equations, equivalent circuit models are insufficient to reflect the mechanical behavior of piezoelectric plates. Electromechanically, fully coupled governing equations of Rosen-type piezoelectric transformers, which are partial differential equations in nature, can be derived to address the deficiencies of the equivalent circuit models. It can be shown that the modal actuator concept can be adopted to optimize the electromechanical coupling effect of the driving section once the added spatial domain design parameters are taken into account, which are three-dimensional spatial dependencies of electromechanical properties. The maximum power transfer condition for a Rosen-type piezoelectric transformer is detailed. Experimental results, which lead us to a series of new design rules, also are presented to prove the validity and effectiveness of the theoretical predictions. PMID- 16382637 TI - Design and analysis of a PZT-based micromachined acoustic sensor with increased sensitivity. AB - The ever-growing applications of lead zirconate titanate (PZT) thin films to sensing devices have given birth to a variety of microsensors. This paper presents the design and theoretical analysis of a PZT-based micro acoustic sensor that uses interdigital electrodes (IDE) and in-plane polarization (IPP) instead of commonly used parallel plate-electrodes (PPE) and through-thickness polarization (TTP). The sensitivity of IDE-based sensors is increased due to the small capacitance of the interdigital capacitor and the large and adjustable electrode spacing. In addition, the sensitivity takes advantage of a large piezoelectric coefficient d33 rather than d31, which is used in PPE-based sensors, resulting in a further improvement in the sensitivity. Laminated beam theory is used to analyze the laminated piezoelectric sensors, and the capacitance of the IDE is deduced by using conformal mapping and partial capacitance techniques. Analytical formulations for predicting the sensitivity of both PPE- and IDE-based microsensors are presented, and factors that influence sensitivity are discussed in detail. Results show that the IDE and IPP can improve the sensitivity significantly. PMID- 16382638 TI - Numerical analysis of lamb wave generation in piezoelectric composite IDT. AB - An equivalent, single-layer model for Lamb wave generation by interdigital transducer (IDT) on composite host structures is developed. The additional complexities generally encountered while launching the surface acoustic wave (SAW) on composite structure, such as the coupling between the Lamb wave modes, the complicated nature of the electromechanical actuation etc. are considered. The model of infinite IDT is extended to deal with the finite IDT with edge discontinuities. The effect of electromechanical actuation on the wavelength shifts with respect to the passive case is investigated. The problem of electrically driven instability within the IDT is analyzed. Numerical results are reported by considering a model of the IDT integrated with the host structure, which shows that there are significant deviations from the conventional design estimates while launching a targeted mode. The proposed approach may enable one to obtain new designs in material systems and geometry that avoid mode-mixing, or to introduce it by choice. PMID- 16382639 TI - Doppler ultrasound spectral enhancement using the Gabor transform-based spectral subtraction. AB - Most of the important clinical indices of blood flow are estimated from the spectrograms of Doppler ultrasound (US) signals. Any noise may degrade the readability of the spectrogram and the precision of the clinical indiCes, so the spectral enhancement plays an important role in Doppler US signal processing. A new Doppler US spectral enhancement method is proposed in this paper and implemented in three main steps: the Gabor transform is used to compute the Gabor coefficients of a Doppler US signal, the spectral subtraction is performed on the magnitude of the Gabor coefficients, and the Gabor expansion with the spectral subtracted Gabor coefficients is used to reconstruct the denoised Doppler US signal. The different analysis and synthesis windows are examined in the Gabor transform and expansion. The signal-to-noise ratio (SNR) improvement together with the overall enhancement of spectrograms are examined on the simulated Doppler US signals from a femoral artery. The results show the denoising method based on the orthogonal-like Gabor expansion achieves the best denoising performance. The experiments on some clinical Doppler US signals from umbilical arteries confirm the superior denoising performance of the new method. PMID- 16382640 TI - Equality of the energy and group velocities of bulk acoustic waves in piezoelectric media. AB - The equality of the energy and group velocities of bulk acoustic waves in a lossless piezoelectric medium is demonstrated, with the energy velocity defined from the generalized energy density and the generalized Poynting vector. PMID- 16382641 TI - Performance of a piezoelectric harvester in thickness-stretch mode of a plate. AB - We studied mechanical-to-electrical power conversion of a piezoelectric plate driven mechanically into thickness-stretch vibrations. We have derived an analytical solution from the three-dimensional equations of linear piezoelectricity that shows the role of each of the physical parameters in determining the performance of such a piezoelectric device, usually measured by the output power density, the power efficiency, or both. Numerical results are included for illustrating the dependence of the device performance upon these physical parameters. PMID- 16382642 TI - The detection properties of ammonia SAW gas sensors based on L-glutamic acid hydrochloride. AB - This study has investigated an improved surface acoustic wave (SAW) ammonia gas sensor based on L-glutamic acid hydrochloride. It presents an excellent reversibility, sensitivity, and repeatability to ammonia. The frequency shift versus ammonia concentration above 40 degrees C was a monotonic function, and the limit of detection of the sensor at 50 degrees C was 80 ppb. PMID- 16382643 TI - Liquid heating effects by SAW streaming on the piezoelectric substrate. AB - When a liquid is placed on a surface acoustic wave (SAW) propagation surface, a longitudinal wave is radiated into the liquid, and the liquid begins to vibrate stream, jet, and atomize. This phenomenon is known as SAW streaming. In this paper, we describe experimental results concerning the temperature of a thin liquid layer during SAW generation. The results reveal that the temperature of the liquid is a function of the SAW amplitude, which is determined by the applied voltage. This means that the liquid temperature can be controlled by the applied voltage. We conclude that a novel microliquid heating system can be realized using the SAW device. PMID- 16382644 TI - Poll finds physicians very wary of pay-for-performance programs. AB - A new poll sheds light on how physician leaders across the country view the rapidly expanding pay-for-performance movement. Some take a dim view; others see it in a positive light. PMID- 16382645 TI - Pay for performance--for whom the bell tolls. AB - Pay -for-performance programs require physician oversight and adherence to high ethical standards if they are truly going to succeed. PMID- 16382646 TI - Physician buy-in is essential for pay for performance. AB - Blue Cross and Blue Shield of Florida is counting on voluntary participation as it rolls out its innovative pay-for-performance program throughout the state. PMID- 16382647 TI - The dark side of P4P. AB - Examine the pitfalls of pay for performance and learn why some physicians and others are deeply concerned about the idea of incentive pay and fear it will have a negative impact on patient care. PMID- 16382648 TI - Keeping score. Scorecards, profiles and report cards rapidly expanding to track physician performance. AB - Not all doctors like them, some even ignore them, but nevertheless the use of physician scorecards is growing and the scores may soon be widely available to the general public. PMID- 16382649 TI - Survey reveals emphasis on tying pay to quality. Physician executive pay increase holds steady near 7 percent. AB - Double-digit increases were seen in three physician executive positions and there are strong indications that the patient safety movement is impacting the way physician executives are paid. PMID- 16382650 TI - Canadians report results of first physician executive compensation survey. PMID- 16382651 TI - Building a culture of service excellence. PMID- 16382652 TI - Implementing an ISO 9001 quality management system in a multispecialty clinic. PMID- 16382653 TI - I drank the Kool-Aid--and learned 24 key management lessons. PMID- 16382654 TI - Dual agency and fiduciary responsibilities in modern medicine. AB - In today's complex medical world, physicians often face difficult decisions about whom they serve first--patients, corporations, insurance companies, the government, etc. PMID- 16382655 TI - Is pay for performance ethical? PMID- 16382656 TI - Resistance to patient safety initiatives. PMID- 16382657 TI - Health care boards can face criminal and civil liabilities. PMID- 16382658 TI - Measuring return on your most valuable asset. PMID- 16382659 TI - Plasticity of rodent and human hair follicle dermal cells: implications for cell therapy and tissue engineering. AB - The dermal components of the hair follicle exhibit a number of stem cell properties, including regenerative potential, roles in wound healing and the ability to produce a functional dermis. Here we examine the stem cell phenomenon of plasticity, focusing on recent observations of in vitro plasticity of dermal papilla and sheath cells, including previously unpublished data of neuronal-like differentiation. We then briefly address the implications of the stem cell potential of hair follicle dermal cells for the field of tissue engineering. PMID- 16382660 TI - Prevalence of male and female pattern hair loss in Maryborough. AB - Maryborough, in central Victoria has an approximate population of 8000 and census data is well matched for Australia overall. Australia has compulsory voting and registration on the electoral roll. To determine the age-related prevalence of balding among men and women in Maryborough we conducted a postal survey of 5000 men and women aged 20 or older, and 427 were invited to attend for examination. Additional data was collected on dandruff, presence of gray hair. Supplementary questionnaires were sent to 340 children aged 5-9, attending a coeducational primary school. 1456 adults (34.1%) responded to the questionnaire. 396 attended for examination. The prevalence of androgenetic alopecia (AGA) increased with advancing age. 98.6% of men had bitemporal recession and severity was significantly associated with vertex and mid-frontal hair loss (p <0.01) but not age (p = 0.06). In all, 64.4% of women had bitemporal hair loss, and similar to men there was a significant association with mid-frontal hair loss (p =0.042) but not age (p =0.467). One hundred and forty children with completed questionnaires were examined. All 72 females and 68 males were assessed as stage 1 on the mid line part and with no bitemporal recession (frequency stage 1 = 100%, 95% CI (confidence interval) 97.4%-100%). A significant but weak positive association existed between presence of gray hair and history of dandruff (p<0.01). The prevalence of mid-frontal hair loss increases with age and affects 57% of women and 73.5% of men aged 80 and over. PMID- 16382661 TI - Clinical diagnosis of common scalp disorders. AB - Scalp skin is unique on the body due to the density of hair follicles and high rate of sebum production. These features make it susceptible to superficial mycotic conditions (dandruff, seborrheic dermatitis, and tinea capitis), parasitic infestation (pediculosis capitis), and inflammatory conditions (psoriasis). Because these scalp conditions share similar clinical manifestations of scaling, inflammation, hair loss, and pruritus, differential diagnosis is critically important. Diagnostic techniques and effective treatment strategies for each of the above conditions will be discussed. PMID- 16382662 TI - The role of sebaceous gland activity and scalp microfloral metabolism in the etiology of seborrheic dermatitis and dandruff. AB - Most common scalp flaking disorders show a strong correlation with sebaceous gland (SG) activity. Early SG activity in the neonate results in microfloral colonization and cradle cap. After maternal hormonal control subsides, there is little SG activity until puberty, when the SG turns on under sex hormone control. When the SG activity increases, the present but low Malassezia population has a new food source and proliferates, resulting in the scalp itching and flaking common to greater than 50% of adults. Dry scalp flaking, dandruff, and seborrheic dermatitis are chronic scalp manifestations of similar etiology differing only in severity. The common etiology is a convergence of three factors: (1) SG secretions, (2) microfloral metabolism, and (3) individual susceptibility. Dandruff and seborrheic dermatitis (D/SD) are more than superficial stratum corneum disorders, including alteration of the epidermis with hyperproliferation, excess lipids, interdigitation of the corneal envelope, and parakeratosis. The pathogenic role of Malassezia in D/SD has recently been elucidated, and is focused on their lipid metabolism. Malassezia restricta and M. globosa require lipids. They degrade sebum, free fatty acids from triglycerides, consume specific saturated fatty acids, and leave behind the unsaturates. Penetration of the modified sebaceous secretions results in inflammation, irritation, and scalp flaking. PMID- 16382663 TI - Product pharmacology and medical actives in achieving therapeutic benefits. PMID- 16382664 TI - A comparison of hair quality and cosmetic acceptance following the use of two anti-dandruff shampoos. AB - Dandruff is a chronic condition of the scalp requiring vigilant treatment to ameliorate the symptoms of scaling and itching. Frequent shampooing is key to controlling these symptoms while contributing to the cosmetic appearance of the hair. The success of the treatment depends not only on the ability of the shampoo to control the dandruff but also on patient compliance engendered by the cosmetic attributes of the shampoo. This double-blind cross-over study enrolled 40 women with mild to moderate dandruff. Following a 1-wk washout with an unmedicated basic cleansing shampoo, all subjects used each of the two study shampoos for 1 wk. Subjects preferred the 1% pyrithione zinc conditioning shampoo over the 2% ketoconazole shampoo by 75% in terms of overall performance. The dermatologist investigator confirmed the subject preference by noting that hair-combing ease, hair smoothness, and hair frizz/flyaway were statistically significantly better in subjects who used the 1% pyrithione zinc conditioning shampoo for 1 wk. This study suggests that hair condition is an important consideration in the treatment of dandruff due to compliance. PMID- 16382665 TI - Hirsutism and the variable response of the pilosebaceous unit to androgen. AB - The pilosebaceous unit (PSU) response to androgen is variable. Certain population of PSU respond to androgen in a distinctive pattern that results in sexual hair development in some, sebaceous gland development in others. Furthermore, androgen excess is variably manifest in women as hirsutism, acne vulgaris, seborrhea, or pattern alopecia. Although sebaceous cells act as intracrine cells, activating pro-hormones to potent androgens that act within the sebocyte, hair follicle metabolism predominantly inactivates testosterone. Androgen action in the sexual hair follicle appears to be mediated by the dermal papilla and possibly, by inducing expression of a specific keratin, hHa7, in the hair medulla. The data do not clearly support a relationship between idiopathic hirsutism, the hirsutism that occurs in the absence of androgen excess, and variations in androgen mechanism of action. Androgens are prominent among the hormones that modulate the biological mechanism regulating the hair cycle. However, the basis for the variable pattern of PSU response to androgen is unclear, as is the basis for the variable development of hirsutism in response to androgen excess and the incomplete reversal of hirsutism by anti-androgen treatment. Improved treatment of hirsutism awaits improved understanding of the nature of the interaction between androgens and other determinants of hair follicle biology. PMID- 16382666 TI - Role of androgen in mesenchymal epithelial interactions in human hair follicle. AB - Human hair follicles, which are distributed in various and specific sites of the body, appear to have an inherited susceptibility for androgen-dependent growth. Beard, axillary, and frontal scalp dermal papilla cells (DPC) were recently shown to possess the characteristics of androgen target cells. These DPC show strong expression of androgen receptors, and the expression of type II 5alpha reductase is restricted to beard and frontal scalp DPC. These findings suggest that DPC mediate the signals of androgen to follicular epithelial cells in a paracrine fashion. We developed an in vitro co-culture system using DPC and keratinocytes (KC) to characterize the mode of androgen action in human hair follicles. Androgen significantly stimulated the proliferation of KC co-cultured with beard DPC, indicating that beard DPC produce androgen-dependent diffusible growth factors. Insulin-like growth factor-I was identified as one of the androgen dependent paracrine growth factors produced by beard DPC. We also identified the inhibitory role of androgen on the growth of KC co-cultured with DPC from androgenetic alopecia (AGA) when the DPC were transfected with an expression vector encoding the androgen receptor. This growth suppression of KC was mediated by transforming growth factor-beta1 (TGF-beta1) derived from DPC of AGA, suggesting that TGF-beta1 is a paracrine mediator for AGA. PMID- 16382667 TI - Beta-endorphin: the forgotten hair follicle melanotropin. AB - The proopiomelanocortin (POMC) gene and protein are expressed principally in the pituitary and brain (e.g., hypothalamus). The POMC gene, protein, and derived peptides, however, can also be detected in the skin. It appears that POMC can also be processed in the skin, a tissue that also expresses the prohormone convertases PC1 and PC2 and 7B2 protein. All POMC peptides may be produced in the skin epidermis, dermis, and adnexa by epithelial cells, melanocytes, and mesenchymal cells (e.g., immune cells, fibroblasts, and endothelial cells), and can also be released from cutaneous sensory nerve endings (cf. Slominski et al, 2004). PMID- 16382668 TI - Female pattern hair loss and its relationship to permanent/cicatricial alopecia: a new perspective. AB - Female pattern hair loss (FPHL) is a common hair disorder of the central scalp. The clinical change in hair density, related to a change in the hair cycle and miniaturization of the hair follicle, is generally considered to be potentially reversible. However, there is now evidence of a permanent hair loss that develops in a subset of women with FPHL. The presence of a perifollicular lymphohistiocytic infiltrate and fibrosis is seen without follicular drop-out in biopsies of women with FPHL and with a notable follicular drop-out in a cicatricial form of this condition (heretofore called cicatricial pattern hair loss) as well as in fibrosing alopecia in a pattern distribution, currently classified as a subset of lichen planopilaris. The potential relationship of these conditions as well as frontal fibrosing alopecia and central centrifugal cicatricial alopecia, two other conditions of permanent hair loss seen primary in women, is discussed. PMID- 16382669 TI - Desmoglein 4 mutations underlie localized autosomal recessive hypotrichosis in humans, mice, and rats. AB - A newly defined form of inherited hair loss, named localized autosomal recessive hypotrichosis (LAH, OMIM 607903), was recently described in the literature and shown to be linked to chromosome 18. A large, intragenic deletion in the desmoglein 4 gene (DSG4) as the underlying mutation in several unrelated families of Pakistani origin. LAH is an autosomal recessive form of hypotrichosis affecting the scalp, trunk, and extremities, and largely sparing the facial, pubic, and axillary hair. Typical hairs are fragile and break easily, leaving short sparse scalp hairs with a characteristic appearance. Using comparative genomics, we also demonstrated that human LAH is allelic with the lanceolate hair (lah) mouse, as well as the lanceolate hair (lah) rat phenotype. Together, these models provide new information about the role of desmosomal cadherins in disease, and serve as in vivo models for functional and mechanistic studies into the role of desmoglein 4 in the skin and hair follicle. PMID- 16382670 TI - Hair transplantation: current concepts and techniques. AB - Because of changes in technique, hair transplanting can now be offered as a reasonable option to more male and female patients who are not responsive to, or likely to benefit from, medical treatment. These changes have also resulted in exceptionally natural-looking results, even after a single session in an alopecic area, or in many individuals with prior and cosmetically unacceptable transplanting results. Current concepts and techniques are described in the article. Possible disadvantages of some of them are also discussed. In particular, the apparent advantages of "megasessions" of 3000 or more grafts per session, "dense packing" of more than 40 follicular units per cm2 and a new method of harvesting single follicular units directly from the donor area (Follicular Unit Extraction), may not be advantageous as they first seem. PMID- 16382671 TI - Human hair keratin-associated proteins. AB - Hair keratin-associated proteins (KAP) are a major component of the hair fiber, and play crucial roles in forming a strong hair shaft through a cross-linked network with keratin intermediate filaments (KIF), which are produced from hair keratins. Recently, the study of human KAP has advanced significantly. So far, five clusters of human KAP genes have been characterized, leading to the identification of more than 80 individual human KAP genes. In situ hybridization studies have demonstrated sequential and spatial expression patterns of these KAP members in differential portions of the hair fiber cortex and cuticle. Furthermore, several human KAP genes have size polymorphisms that are mainly because of variable numbers of cysteine-rich repeat segments, and the patterns of some of these size variants are distinct between different human populations. PMID- 16382672 TI - Integral lipid in human hair follicle. AB - On the hair surface, cells are covered with a thin lipid layer (LL) covalently bonded to hair proteins. This integral hair lipid is different from sebaceus lipid. We conducted this study to examine the lipid distribution in human hair follicle. Transmission electron microscopy was performed to observe the ultrastructure of the LL. Hair follicles were cut and observed longitudinally along the hair axis. For transmission electron microscopy, new fixative (Lee's fixative: composed of OsO4 and RuO4) was designed as the conventional fixatives such as OsO4 or RuO4 alone were not appropriate for staining for hair follicle lipid. In addition, we measured the chemical composition of integral hair lipid by high-performance thin-layer chromatography. From the above experimental procedure, it was discovered that the lipid in the hair follicle was mainly distributed in hair cuticle and keratinized inner root sheath. A multitude of lamellar granule is observed in the vicinity of the above LL. The chemical composition of integral hair lipid was different from those of epidermal or sebaceous lipids. It is assumed that the LL in the hair follicle is similar to the epidermal LL playing an important role as a skin barrier in the stratum corneum. We proposed here the new terminology "hair barrier" from this point of view. PMID- 16382673 TI - Epididymal cysteine-rich secretory protein 1 encoding gene is expressed in murine hair follicles and downregulated in mice overexpressing Hoxc13. AB - Members of the Hox gene family of transcriptional regulators are believed to play essential roles in hair follicle differentiation, although little is known about the molecular mechanisms mediating these putative control functions. Transgenic mice overexpressing Hoxc13 in hair follicles (GC13 mice) exhibit complex phenotypic alterations including hair shaft defects and alopecia, as well as severe epidermal abnormalities. To identify some of the genetic pathways affected by Hoxc13 overexpression in hair, we performed large-scale differential gene expression analysis on the skin of 5-d GC13 versus normal FVB mice using DNA chip assays. A surprising result of these experiments was the identification of the epididymal cysteine-rich secretory protein 1 (Crisp1) gene as one of the genes with the greatest expression differential, in this case with greater than 20-fold downregulation in skin from GC13 mice. Crisp1 encodes a secreted protein that has originally been found to be abundantly expressed in the epididymis, where it plays a role in sperm maturation. We have localized Crisp1 mRNA in 5-d postnatal murine scapular skin by in situ hybridization, showing its expression to be restricted to the medulla of the hair shaft. Furthermore, we provide evidence for specific interaction of Hoxc13 with at least one cognate binding site found in the Crisp1 promoter region, thus supporting the concept of a Hoxc13/Crisp1 regulatory relationship. In summary, these data establish the hair as a novel site for Crisp1 expression where its functional role remains to be determined. PMID- 16382674 TI - Substantial sex-dependent differences in the response of human scalp hair follicles to estrogen stimulation in vitro advocate gender-tailored management of female versus male pattern balding. AB - In this study, it was investigated how estrogens (17-beta-estradiol, E2) affect the estrogen receptor (ER) expression and gene regulation of male versus female human scalp hair follicles in vitro. Anagen VI follicles from frontotemporal scalp skin were microdissected and organ-cultured for up to 9 d in the presence of E2 (1-100 nm). Immunohistochemistry was performed for ERbeta-expression, known to be predominant in human scalp hair follicles, and for TGF-beta2-expression (as negative key hair growth modulator), and E2-responsive genes in organ-cultured human scalp hair follicles (48 h, 10 nM) were explored by cDNA microarray, using a commercial skin focus chip (Memorec, Cologne, Germany). The distribution pattern of ERbeta and TGF-beta2-immunoreactivity differed between male and female hair follicles after 48 h culture. Of 1300 genes tested, several genes were regulated sex-dependent differently. The study reveals substantial sex-dependent differences in the response of frontotemporal human scalp hair follicles to E2. Recognition and systematic dissection of the E2-dependent gene regulation will be crucial for the development of more effective, gender-tailored management strategies for female versus male pattern balding. PMID- 16382676 TI - Follicular targeting--a promising tool in selective dermatotherapy. AB - The penetration of topically applied compounds varies considerably in the different regions of the human body. The presence of hair follicles significantly contributes to this effect by an increase in surface area and a disruption of the epidermal barrier towards the lower parts of the hair follicle. The human hair follicle, hereby, serves not only as a reservoir, but also as a major entry point for topically applied compounds. Topical delivery of active compounds to specific targets within the skin may help reduce side-effects caused by unspecific reactions, and may help develop new strategies in the prevention and treatment of skin diseases. Various drug carrier and drug delivery systems are currently being investigated. The aim of these investigational efforts is to direct topically applied compounds to the different types of hair follicles and, ideally, to specific compartments and cell populations within the hair follicles. Follicular targeting offers opportunities for new developments, not only in hair therapy and in the treatment of hair follicle associated diseases but also in gene therapy and immunotherapy. PMID- 16382675 TI - Edar signaling in the control of hair follicle development. AB - Ectodysplasin receptor Edar and its ligand Eda A1, as well as their related receptor Xedar and ligand Eda A2, are recently discovered members of the tumor necrosis factor superfamily that signal predominantly through the nuclear factor kappaB and c-jun N-terminal kinases pathways. Mutations in genes that encode proteins involved in Edar signaling pathway cause hypohidrotic ectodermal displasias in humans and mice and characterized by severe defects in development of ectodermal appendages including hairs, teeth, and exocrine glands. Here, we summarize the current knowledge of molecular mechanisms underlying the involvement of Edar signaling pathway in controlling hair follicle (HF) development and cycling. Genetic and experimental studies suggest that Edar signaling is involved in the control of cell fate decision in embryonic epidermis, as well as in the regulation of cell differentiation programs in the HF. Loss or gain of Edar signaling affects the initiation of several HF types (guard and zig-zag HF), hair shaft formation, as well as sebaceous gland morphology. We also review data on the cross-talk between Edar and Wnt, transforming growth factor-beta/bone morphogenic protein/activin, and Shh signaling pathways in the control of HF development and cycling. PMID- 16382677 TI - Dermatopathology workshop summary, Berlin 2004. PMID- 16382678 TI - Analysis of hair follicles in mutant laboratory mice. AB - Hair follicles, or pilosebacous units, are complicated anatomic structures. Analysis of abnormalities of these structures can be difficult without adequate knowledge of normal changes associated with embryonic and postnatal development as well as regular cycling. A variety of systematic approaches to analysis of the pilosebacous unit are provided here. PMID- 16382679 TI - Epilation today: physiology of the hair follicle and clinical photo-epilation. AB - Despite the variations of length and type of hair (vellus or terminal), the growth of human hair in all body sites is cyclic. Phases of active hair growth, or anagen, are separated by periods of quiescence, or telogen. The duration of both phases varies greatly depending on the body site. Whether hairs are in anagen/telogen at the time of hair removal is important because only anagen hairs are particularly sensible to physical insults. Photo-epilation is a technique for long-term removal of unwanted hair by thermal destruction of the hair follicle and its reproductive system (stems cells). As melanin is the main chromophor existing in hair follicles the corresponding wavelength spectrum would range from ultraviolet up to infrared light. Furthermore longer wavelengths are preferred as the cromophor lies deep in the skin and the penetration of light is increasing with the wavelength. Thus, in the range of 600-1100 nm melanin absorption may be used for selective photothermolysis of hair follicles. Yet to be resolved questions for permanent destruction are the location of the key follicular target and the possible influence of the hair growth cycle on photothermolysis-induced hair removal. An overview on the individual physiology of the hair follicle is given to discuss the latest strategies for photo-epilation. PMID- 16382681 TI - Proceedings of the 4th International Meeting of Hair Research Societies. June 17 19, 2004. Berlin, Germany. PMID- 16382680 TI - Hair pigmentation: a research update. AB - Hair is a uniquely mammalian trait with important functions, most easily appreciated in furred mammals. Our skin and hair color contribute very significantly to our overall visual appearance by highlighting striking variations between human sub-groups. Although melanins, hemoglobins, and carotenoids define the color perceived at the skin surface, our hair color relies only on the presence or absence of different melanins. The hair shaft's physical aspects provide only minor color modification. Various selective evolutionary pressures have determined that within the context of our specific ethnic backgrounds a bewildering array of natural shades are seen; ranging from yellows, reds, and browns to black and that harbinger of lost youth, gray/white hair. Skin/hair follicle melanins are formed in cytoplasmic organelles called melano somes produced by neural crest-derived pigment cells called melano-cytes and are the product of a complex, phylogenetically ancient, biochemical pathway called melanogenesis. The following provides a review of research presented at the 4th Intercontinental Meeting of Hair Research Societies 2004 and so is not intended to represent a fully comprehensive overview of the subject-for that readers are directed to key references. PMID- 16382682 TI - Recent findings with computerized methods for scalp hair growth measurements. AB - Sensitive tools have been developed in order to monitor hair loss and treatment responses. Recently the Tricho-Scan was presented (by RH) as such a method which combines epiluminescence microscopy (ELM) with automatic digital image analysis. Herewith new TrichoScan data obtained from 10 women and 21 men with androgenetic hair loss after 6 mo of treatment with 5%-minoxidil are presented. Even in this small cohort of patients, we noticed a significant increase of hair density, cumulative hair thickness and terminal hair counts. Alternative methods were developed during a human alopecia investigation and research technology (HAIR Technology) programme at Skinterface. This involves contrast-enhancement, image acquisition, and processing by qualified technicians followed by computer assisted image analysis. The specific identification of exogen hair, further adds to this very refined non-invasive investigative method for hair follicle function investigation. Regional variations of hair growth dynamics do exist in the human scalp such as in female patients complaining of hair loss, scalp hair density and growth on top of the head differs significantly from the occipital site. Finally, from transversal studies and from detailed monitoring of subsequent hair cycles during longitudinal studies, data were obtained that support the fact that shortening of hair cycle, slowing down of growth rates and thinning of hair shafts are heralding hair miniaturisation. In the workshop the TrichoScan, the method of Canfield and Skinterface have been shown. PMID- 16382683 TI - Dermocosmetic aspects of hair and scalp. AB - Hair cosmetic agents are preparations intended for placing in contact with the hair and scalp, with the purpose of cleansing, promoting attractiveness, altering appearance, and/or protecting them in order to maintain them in good condition. Current shampoo formulations and styling products are tailored to the variations associated with age, gender, hair quality, hair care habit, and specific problems relating to the superficial condition of the scalp. The great amount of variables to be accounted for, some of them contradictory and hard to bring into accord, makes adequate product formulation a challenge and requires continuous research. Recent hair cosmetic developments include topical hair growth stimulants, photoprotectors, and anti-aging compounds. The advances in molecular technology have opened the avenue to the study of the molecular basis of hair growth and its deviations, as well as of the aging process. The discovery of potential pharmacological targets and the development of selective and effective delivery systems following topical application indicate further strategies for maintenance of healthy hair and scalp in the young and old. PMID- 16382684 TI - Effectiveness of finasteride on patients with male pattern baldness who have different androgen receptor gene polymorphism. AB - Finasteride is not necessarily effective on all of the male pattern baldness (MPB) patients. To know any factor which correlates with the effectiveness of finasteride, the polymorphism of androgen receptor (AR) gene was analyzed. Symptoms of the 488 MPB patients (18-62 y) before and after treatment with total dose of 10 mg or more of finasteride was typed by photographic method. The number of CAG and GGC repeats in AR gene of MPB patients was determined by DNA sequencing. When the number of the triplet repeats (CAG + GGC) was plotted against the degree of symptom improvement after treatment with this drug, a broad correlation between these variables was observed. The smaller the repeat number, the higher the improvement with finasteride. The group of patients with shorter repeat region in AR gene responded better to this drug than that with longer repeat region, although the former patients tended to reveal severe initial symptoms. Determination of such polymorphism is thought to be useful in the drug choice for MPB patients. PMID- 16382685 TI - Three etiologic facets of dandruff and seborrheic dermatitis: Malassezia fungi, sebaceous lipids, and individual sensitivity. AB - Application of new molecular and biochemical tools has greatly increased our understanding of the organisms, mechanisms, and treatments of dandruff and seborrheic dermatitis. Dandruff results from at least three etiologic factors: Malassezia fungi, sebaceous secretions, and individual sensitivity. While Malassezia (formerly P. ovale) has long been a suspected cause, implicated by its presence on skin and lipophylic nature, lack of correlation between Malassezia number and the presence and severity of dandruff has remained perplexing. We have previously identified the Malassezia species correlating to dandruff and seborrheic dermatitis. In this report, we show that dandruff is mediated by Malassezia metabolites, specifically irritating free fatty acids released from sebaceous triglycerides. Investigation of the toxic Malassezia free fatty acid metabolites (represented by oleic acid) reveals the component of individual susceptibility. Malassezia metabolism results in increased levels of scalp free fatty acids. Of the three etiologic factors implicated in dandruff, Malassezia, sebaceous triglycerides, and individual susceptibility, Malassezia are the easiest to control. Pyrithione zinc kills Malassezia and all other fungi, and is highly effective against the Malassezia species actually found on scalp. Reduction in fungi reduces free fatty acids, thereby reducing scalp flaking and itch. PMID- 16382686 TI - Hairstyles in the arts of Greek and Roman antiquity. AB - Styling one's hair seems to be an innate desire of humans to emphasize their beauty and power. As reviewed here, hairstyles were influenced by preceding cultures, by religion, by those depicted for gods and emperors on sculptures and coins. In addition, they were determined by aspects of lifestyle such as sports, wealth, and the desire to display inner feelings. The historical changes in fashions can be exemplarily followed by a visitor to an art collection of Graeco Roman antiquity. The study of hairstyles permits an insight into very basic aspects of the self-conception of individuals and of the respective societies. PMID- 16382687 TI - Follicular penetration and targeting. AB - In the past, intercellular penetration was assumed to be the most important penetration pathway of topically applied substances. First hints that follicular penetration needs to be taken into consideration were confirmed by recent investigations, presented during the workshop "Follicular Penetration and Targeting" at the 4th Intercontinental Meeting of Hair Research Societies", in Berlin 2004. Hair follicles represent an efficient reservoir for the penetration of topically applied substances with subsequent targeting of distinct cell populations, e.g., nestin-expressing follicular bulge cells. The volume of this reservoir can be determined by differential stripping technology. The follicular penetration processes are significantly influenced by the state of the follicular infundibulum; recent experimental investigations could demonstrate that it is essential to distinguish between open and closed hair follicles. Topically applied substances can only penetrate into open hair follicle. Knowledge of follicular penetration is of high clinical relevance for functional targeting of distinct follicular regions. Human hair follicles show a hair-cycle-dependent variation of the dense neuronal and vascular network. Moreover, during hair follicle cycling with initiation of anagen, newly formed vessels occur. Thus, the potential of nestin-expressing hair follicle stem cells to form neurons and blood vessels was investigated. PMID- 16382688 TI - [The Hippocratic Oath: how many covenants, how many oaths?]. AB - This study focuses on the ancient text of the Hippocratic Oath. Ancient authors refer to various oaths in the context of medical practice, but none of these closely resemble the Hippocratic Oath. The text of the Hippocratic Oath contains several contractual clauses, which clearly reflect a strictly structured community within an institutionalized professional framework. Comparison with the epigraphic and papyrological tradition suggests placing the text in the Hellenistic-Roman period. In particular, investigation of the oldest remaining ancient textual witness, the POxy XXXI 2547 of the third century A.D., reveals Christian influences in the use of language. This indicates a stage of development of the Hippocratic Oath in which the latter existed within a Christian context. The article concludes that, as early as the third century A.D., Christian influences played an equally significant role in the transmission of the Oath as pagan influences. PMID- 16382689 TI - [Strategies of medical self-authorization in early modern medicine: the example of Volcher Coiter (1534-1576)]. AB - Based on the example of Volcher Coiter--a town physician at Nuremberg and one of the leading anatomists in early modern medicine--, this essay points out that the authoritative status of contemporary physicians mainly was predicated on an interplay of self-fashioning and outside perception. It provides ample evidence that Coiter made use of several characteristic rhetorical and discourse-related strategies of self-authorisation such as the participation in social networks, a highly convincing technique of self-fashioning by emphasizing particular erudition, the presentation of academic medicine as a science authorised by god and the concurrent devaluation of non-academic healers. Furthermore, graphic and visual strategies of self-authorisation could be ascertained: Coiter took care for a premium typography of his books. He also used his talent as a graphic artist in his books to visualise his medical concepts. Moreover, the so-called 'Nuremberg Portrait' of Coiter served to illustrate his outstanding authority. PMID- 16382690 TI - A Danish provincial physician and his patients; the patient records from the practice of Christopher Detlev Hahn in Aarhus around 1800. AB - The Danish physician Christopher Detlev Hahn (1744-1822) graduated in medicine from Halle and set up a medical practice in Aarhus in 1766. More than 2000 of his patient records, bound in twenty-seven volumes, are kept in the archives of the Medical Museum in Copenhagen. The records, which are written in Latin and resemble hospital records, contain entries from each visit to the patients. A sample of these records, especially those from 1806, have been analysed regarding diagnoses, alleged causes of disease, examination of patients and prescription of different treatments. It was noticeable to what extent Hahn, who belonged to the Hippocratic tradition, individualised both diagnosis and treatment. The introduction of variolation and later vaccination is discussed. This collection of records provides a unique insight into medical practice at that time outside the hospitals. PMID- 16382691 TI - [Mild radium therapy: radiation medicine-, life-, body- and personal hygiene remedies in the first half of the 20th century]. AB - During the first half of the 20th century numerous drugs, foodstuffs and cosmetics were brought on the market, whose supposed effects were explained with their weak radioactivity. Their subtle radiation was believed to stimulate the vital forces of the body, thus leading to recovery from illness, or to an improvement in beauty and to rejuvenation. Among others, bath and drinking waters enriched with radioactive materials were advertised for this purpose. The then known radioactive medicines included preparations of healing earth, the so-called Salus-Oil, the TRUW preparations, and "Radithor", which was popular in the United States. There were also radioactive foodstuffs (butter, chocolate, rusk) and cosmetics. This mild radiotherapy may be characterised as a form of bio dynamistic healing. PMID- 16382692 TI - [On the origins of the Druckrey-Kupfmuller Papers (1948-1949): dose-effect relations in carcinogenic substances]. AB - Theoretical knowledge about the relationship between drug dosage and tissue response in pharmacology and toxicology in general, and concerning the action of carcinogenic substances in particular, largely dates back to two papers by Hermann Druckrey and Karl Kupfmuller, published in 1948 and 1949, respectively. The definition of cumulative poisons and the first proof of the irreversibility of carcinogenic effects had an extraordinary impact on cancer research. The story of how these papers came to be written is remarkable for several reasons. First, an interdisciplinary approach resulted in a landmark interpretation of data regarding the quantitative analysis of cancer development. Second, an intensive collaboration developed between Druckrey, a cancer researcher, and Kupfmuller, a mathematically adept electrical and communications engineer. Third, during their temporary "full-time co-operation", both researchers were in an allied internment camp. Thus, their fundamental scientific achievement ultimately resulted from the fulfilment of three conditions of innovation, i.e. compatible visions of the co operating scientists, interdisciplinary links, and chance. PMID- 16382693 TI - Immunodeficiency: nobody is immune. PMID- 16382694 TI - Autoimmune lymphoproliferative syndrome. PMID- 16382695 TI - Intravenous immunogloblins: myth and reality. PMID- 16382696 TI - Wegener's granulomatosis: a view from the granulomatous side of the disease. AB - Although the airway granulomata in Wegener's granulomatosis were stressed initially by Friedrich Wegener himself, in the last few decades systemic lesions mainly caused by acute vasculitis have received the most attention. However, recently, the implication of granulomatous manifestations in WG has raised much interest. The present data suggest that an aberrant Th1-type response might play a role in the initiation of WG, clinico-pathologically characterized by granulomatous inflammation rather than vasculitis. Disease progression to generalized WG with the predominance of vasculitic manifestations is associated with a "switch" or further complexity of the collective T cell response with the appearance of another subset of Th2-type cells and a less prominent Th1-type cytokine production in the granulomatous lesions of the upper respiratory tract. However, the clinical significance of the granulomatous inflammation is not yet completely understood. Further research will also have to focus on the role of the granulomata during relapsing disease. We review present knowledge of granulomatous inflammation in WG. Morphologic aspects, the scale of cytokine alterations as well as the variety of clinical manifestations are discussed. PMID- 16382697 TI - Chronic urticaria: an evolving story. PMID- 16382698 TI - Low levels of heat shock proteins-60 and -65 autoantibodies in Sjogren's syndrome. AB - BACKGROUND: Heat shock proteins are highly conserved immunodominant antigens found in various species. Humoral immune responses to mycobacterial HSP65 and human HSP60 have been established in a number of human autoimmune diseases. OBJECTIVE: To assess the prevalence of antibodies to HSP60 kDa and HSP65 kDa in patients with Sjogren's syndrome as compared to normal subjects. METHODS: Thirty seven patients with SS were compared with normal controls. The antibodies against human HSP60 were measured by the Anti-Human (IgG/IgM) HSP60 ELISA kit. IgGs and IgMs to mycobacterial HSP65 were determined using an enzyme-linked immunosorbent assay with mycobacterial recombinant HSP65 antigens. RESULTS: The levels of both anti-HSP60 and -HSP65 were lower in patients compared with controls. IgG autoantibodies to HSP60 were significantly different between groups: 162 +/- 55.1 ng/ml in controls versus 112.3 +/- 30.6 ng/ml in SS patients (P < 0.001). The levels among controls of anti-HSP65 IgM isotype were also significantly higher than among the SS patients: 111.6 +/- 33.4 U/ml versus 96.1 +/- 8.9 U/ml (P= 0.01). CONCLUSIONS: The results of the present study show that the levels of different isotypes of anti- HSP60 and HSP65 antibodies were lower in patients with SS than in normal subjects. Additional studies in larger patient populations are required to evaluate the prevalence of these autoantibodies in SS patients. PMID- 16382699 TI - Bronchial hyperactivity, sputum analysis and skin prick test to inhalant allergens in patients with symptomatic food hypersensitivity. AB - BACKGROUND: Dyspnea may be a presenting symptom of type I food hypersensitivity; bronchial hyperactivity, without known asthma, can coexist in patients with food allergy. OBJECTIVE: To evaluate airway involvement in young adult patients with food allergy and no asthma and compare the findings to those of patients with food allergy and asthma, with food allergy and allergic rhinitis, with asthma and no food allergy, and of apparently healthy controls. METHODS: The evaluation involved prick skin test to food (65 allergens) and inhalants (24 allergens), spirometry, methacholine inhalation challenge, and induced sputum for cell analysis. The five groups consisted of 18 patients with food allergy alone, 11 with food allergy and asthma, 13 with food allergy and allergic rhinitis, 10 with asthma alone, and 10 controls. RESULTS: Patients with food allergy alone were mainly (86%) skin sensitive to pollens. Those with either asthma or allergic rhinitis were mainly (95%) sensitive to mites. BHR was detected in 40% of the patients with food allergy alone, 55% of the patients with allergic rhinitis, and 100% of the patients with asthma. Cell counts in the sputum of patients with asthma and in those with food allergy and asthma showed higher eosinophil counts compared to those with food allergy and allergic rhinitis. Patients with food allergy and no asthma, regardless of BHR status, had mainly neutrophils in the sputum. CONCLUSIONS: Patients with food allergy are highly likely to have concomitant asymptomatic BHR. Mite sensitivity in patients with food allergy predicts respiratory allergy (either asthma or allergic rhinitis). High eosinophil levels in the sputum of food allergy patients predict respiratory involvement. PMID- 16382700 TI - Hospital admission trends for pediatric asthma: results of a 10 year survey in Israel. AB - BACKGROUND: In the last decade there has been an increase in asthma morbidity. Hospital admission rates for childhood asthma are influenced by the prevalence of asthma and the quality of asthma care. OBJECTIVES: To assess trends in hospital admission and readmission rates for childhood asthma in the Jezreel Valley in Israel in the last decade, and to evaluate the possible effect of changes in asthma treatment on hospitalization for acute asthma during this period. METHODS: All records of pediatric patients from the central hospital in the Jezreel Valley in northeastern Israel over a 10 year period from 1990 through 1999 who were diagnosed as having asthma were thoroughly reviewed and analyzed for admissions, re-admissions, and treatment before and during admissions RESULTS: There were 1584 admissions, 1208 were first-time admissions and 376 were re-admissions. The number of first-time admissions increased significantly over time (P < 0.0001), with a significant decrease in re-admissions (P < 0.005); this finding was more significant in children under the age of 8 years (P < 0.005). The length of hospital stay decreased significantly from 3.3 days to 2.7 days (P < 0.002). Significant changes in the use of medications included an increase in inhalant glucocorticoids and a decrease in the use of sodium cromoglycate and theophylline. Controller medication use was concomitant with a significant decrease in the re-admission rates. CONCLUSIONS: The increase in the admission rate and the decrease in the rate of re-admissions and the length of hospital stay probably reflect the increase in the prevalence of asthma and changes in its treatment, respectively. It is essential that asthma be recognized as a significant cause of morbidity and that controller medications be administered to decrease its severity, morbidity, and resultant hospital admissions. PMID- 16382701 TI - Cross-reactive hypersensitivity to COX inhibitors in a child with mild allergic rhinitis. PMID- 16382702 TI - Effect of montelukast on basophil releasability in patients with asthma. AB - BACKGROUND: The anti-inflammatory effect of montelukast, a leukotriene receptor antagonist, in patients with bronchial asthma is not entirely clear. Basophils can release a variety of mediators, including histamine and leukotriens, which most likely play an active role in the late allergic response. OBJECTIVES: To study the effect of montelukast (10 mg/day) on histamine and cysteinyl leukotriene release from basophils taken from 12 mild atopic asthmatic patients who took the drug for 4 weeks. METHODS: Basophils were withdrawn at baseline, and after 48 hours, 1 week, and 4 weeks of therapy. Histamine was measured by a radioenzymatic method and leukotrienes by immunologic assay. Histamine and cysLT release was measured spontaneously and following stimulation with interleukin-3 and anti-immunoglobulin E. Spirometry and symptom score were measured before and during treatment. RESULTS: During the treatment with montelukast there were no significant changes in spontaneous, IL-3 and anti-IgE-induced histamine release. cysLT release decreased significantly only after 4 weeks of treatment (from 2899 +/- 550 pg/ml at baseline to 2225 +/- 430 pg/ml at 4 weeks, P= 0.02). CONCLUSIONS: Montelukast does not seem to affect the release of histamine from basophils but mildly inhibits the cysLT release seen after 4 weeks of treatment. PMID- 16382703 TI - Camel milk for food allergies in children. AB - BACKGROUND: Food allergies in children are often very serious and can lead to anaphylactic reactions. Observations that camel milk ameliorates allergic reactions were noted over the years. The effect of camel milk is probably related to its special composition. OBJECTIVES: To investigate the effect of camel milk in several children with severe food (mainly milk) allergies. METHODS: We studied eight children with food allergies who did not benefit from conventional treatment. Their parents, or their physicians, decided to try camel milk as a last resort. The parents were advised by the authors - who have considerable experience with the use of camel milk - regarding how much and when the children should drink the milk. The parents reported daily on the progress of their children. RESULTS: All eight children in this study reacted well to the milk and recovered fully from their allergies. CONCLUSIONS: These encouraging results should be validated by large-scale clinical trials. PMID- 16382704 TI - Serum immunoglobulin E levels in Israeli-Ethiopian children: environment and genetics. AB - BACKGROUND: Since 1984, several waves of Ethiopian immigrants have settled in Israel. On arrival they were found to be highly infected with intestinal parasites and to have increased serum immunoglobulin E and eosinophilia. OBJECTIVES: To study serum IgE levels in Ethiopian children growing up in the environment of Israel. METHODS: We assessed four groups of children of Ethiopian origin: a) adolescents examined on their arrival to Israel (group 1, n=11); b) adolescents born in Ethiopia and living in Israel for more than 7 years (group 2, n=10); c) children of Ethiopian origin born in Israel, without a history of allergy or asthma (group 3, n=15); and d) asthmatic children of Ethiopian origin born in Israel (group 4, n=8). A thorough clinical interview and examination as well as laboratory work up (including serum IgE levels, stool parasites and absolute eosinophil count) were performed. RESULTS: Group 1 (11 newly arrived Ethiopian adolescents) had a mean eosinophil count of 688 cells/ml (0-1739) and a mean serum IgE of 1043 IU/ml (253-2932), P< 0.0009 as compared to group 2. Helminthic parasites were observed in 8/11 individuals; after 1 year of follow-up and anti-parasitic treatment, serum IgE levels did not change significantly. Group 2 (10 Ethiopian born adolescents living in Israel for on average 10 years, 7-15 years) had a normal leukocyte count, MEC 192 cells/ml (range 54-289), serum IgE 142 IU/ml (range 14-399 IU/ml) and no parasites in stool. Group 3 (15 Ethiopian children born in Israel) had a normal leukocyte count, MEC 128 cells/ml (0-324), serum IgE 55 IU/ml (7-189 IU/ml), similar to age-matched Israeli controls. In group 4 (8 Israeli born children of Ethiopian descent diagnosed with asthma), serum IgE showed significant elevation compared to Israeli age-matched asthmatic children (P< 0.005). CONCLUSIONS: High levels of IgE found in Ethiopian children on arrival to Israel declined to Israeli control levels after several years of living in the new environment. Ethiopian children born in Israel had normal levels of IgE, suggesting that environment is the main factor affecting IgE levels in this population. Israeli born Ethiopian children with asthma had significantly increased serum IgE levels compared to asthmatics of Israeli origin. These findings suggest that both environmental and genetic factors determine the level of serum IgE in these children. PMID- 16382705 TI - Psychiatric admissions and hospitalization in Israel: an epidemiologic study of where we stand today and where we are going. AB - BACKGROUND: Analysis of the trends in psychiatric admissions and discharges is necessary to correctly plan and distribute resources, especially given the current international climate of "deinstitutionalization." Israel, too, is implementing "reform" in the national psychiatric system - to transfer psychiatric treatment from a hospital to a community setting. OBJECTIVES: To analyze admission and discharge patterns, explore trends in psychiatric hospital length of stay, and compare these characteristics between first-episode and chronic patients, between children, youth and adults, and between hospitals. METHODS: All admissions and discharges from inpatient psychiatric wards between the years 2000 and 2004 were analyzed and characterized according to age, length of hospitalization, legal status, and nature of admitting institution (state hospital, health fund, general hospital). RESULTS: Mean length of stay in adults decreased during the 5 year study period, from 37.6 days in 2000 to 36.4 days in 2004. In years with higher admissions, hospital stay was shorter (P < 0.05). Length of stay in psychiatric wards in general hospitals was shorter than in state hospitals (P < 0.001). In contrast to adults and children, length of stay among adolescents showed a gradual increase (P < 0.05). Involuntary hospitalization comprised 25.3% of all admissions, and 16.8% of discharged patients were re-admitted within 30 days. A dramatic decrease (24.3%) in the number of chronic hospitalizations was noted. CONCLUSIONS: Various factors may account for these developments. Protracted hospitalizations may be reduced through changes in various aspects of treatment planning and psychiatric care continuum. The decrease in number of admissions, length of stay and number of chronically admitted patients remains in line with international practices. Particular attention needs to be devoted to planning and funding so that availability of community services matches reduction in psychiatric hospitalization. PMID- 16382706 TI - Thrombophilia: a risk factor for cerebral palsy? AB - BACKGROUND: The cause of cerebral palsy remains unknown in most cases. Factor V Leiden mutation, a common cause of hereditary thrombophilia, has been associated with CP. OBJECTIVES: To analyze the prevalence of factor V Leiden (G1691A), prothrombin (G20210A), and methylenetetrahydrofolate reductase (C677T) mutations in children with CP. METHODS: Sixty-one Jewish and Arab children with CP were studied for the presence of the three gene mutations associated with thrombophilia. RESULTS: We found that 41% of the children with CP and 33% of the controls carry one or more of the studied mutations (P = 0.348). The prevalence of the factor V mutation was 27.9% in CP and 16.4% in controls (P= 0.127). The frequency of the other two genetic factors was even less significant. The FVL mutation was found in 35% of the Arab CP patients (15/42) and in 22% of the controls from the same population (9/40) (P= 0.067). CONCLUSIONS: Each of the genetic factors studied was shown to be related to CP. Despite the high frequency of FVL among the studied patients, we were unable to prove a significant correlation between FVL and CP, mainly because this factor is frequent in the Arab control group. In this population a trend toward significance can be seen (P= 0.067). Larger studies are needed to validate the significance of these results. PMID- 16382707 TI - Intravenous chlorpromazine for the emergency treatment of uncontrolled symptomatic hypertension in the pre-hospital setting: data from 500 consecutive cases. AB - BACKGROUND: Chlorpromazine is a dopamine-receptor antagonist antipsychotic agent. Because of its strong alpha-blocking and sedative actions, it has also been used as emergency therapy for extreme arterial hypertension. Published reports to date have included very small numbers of patients (i.e., 5-30). OBJECTIVES: To analyze data on almost 500 patients who received intravenous chlorpromazine for the emergency treatment of uncontrolled symptomatic hypertension in the pre-hospital setting. METHODS: We reviewed data from 496 consecutive patients who received intravenous chlorpromazine as emergency therapy for uncontrolled symptomatic hypertension. Chlorpromazine was injected intravenously. The dose was 1 mg every 2-5 minutes until the systolic pressure was < or =140 mmHg and the diastolic pressure < or =100 mmHg with alleviation of symptoms. RESULTS: The mean dose of chlorpromazine administered was 4.5 +/- 5 mg (range 1-50 mg). Only 33 patients (7%) required >10 mg. Chlorpromazine reduced systolic BP from 222.82 +/- 26.31 to 164.93 +/- 22.66 mmHg (P< 0.001) and diastolic BP from 113.5 +/-16.63 to 85.83 +/ 11.61 mmHg (P< 0.001). The sinus rate decreased from 97.9 +/- 23.5 to 92.2 +/- 19.7 beats per minute (P< 0.001). These results were achieved within the first 37 +/- 11 minutes. CONCLUSIONS: Intravenous chlorpromazine is safe and effective when used as emergency treatment for uncontrolled symptomatic hypertension. PMID- 16382708 TI - Intensive cholesterol lowering: no longer a myth. PMID- 16382709 TI - The Israeli reform of psychiatric hospitalizations. PMID- 16382710 TI - New treatment approaches in multiple myeloma. PMID- 16382711 TI - Hemorrhagic complications of low molecular weight heparins (LMWHs). PMID- 16382712 TI - Using fibrin glue in endonasal surgery. PMID- 16382713 TI - Complementary and alternative Medicine (CAM) diagnostics. PMID- 16382714 TI - [The road travelled]. PMID- 16382715 TI - [Screening of the prostate cancer. "lnstantaneous" of a daily practice among 300 general practitioners of the canton of Vaud]. AB - This investigation brings information on the practice of 300 general practitioners of the canton of Vaud as for the use of the prostate-specific antigen (PSA) and the digital rectal examination (DRE) in the prostate cancer screening. The high rate of answer shows their interest for this problem which they deal with in adequacy with the international recommendations published by the societies of urology. PMID- 16382716 TI - [Biochemical failure after curative treatment for localized prostate cancer]. AB - Biochemical failure after curative treatment for localized prostate cancer is frequent. The diagnosis of biochemical failure is clear when PSA levels rise after radical prostatectomy, but may be more difficult after external beam radiation therapy. The main difficulty once biochemical failure is diagnosed is to distinguish between local and distant failure, given the low sensitivity of standard work-up exams. Metabolic imaging techniques currently under evaluation may in the future help us to localize the site of failures. There are several therapeutic options depending on the initial curative treatment, each with morbidity risks that should be considered in multidisciplinary decision-making. PMID- 16382717 TI - [The therapeutic dilemma of the early invasive bladder cancer]. AB - Transitional tumours of the bladder are classically divided in superficial (lesions of the mucosa and the submucosa) and invasive (infiltrating the detrusor) cancers. However, the recurrence and progression rate of submucosal urothelial cancers is highly variable. Therefore, management of such neoplasias is very challenging, some patients requiring a cystectomy, whereas others can be managed less invasively with endoscopic resection eventually associated with intravesical chemotherapy. We review herein the prognostic factors which help us to orientate our patients. We also emphasize the importance of the subclassification of the micro-invasive stage, and its practicability, which is reliable and simple, as opposed to the general belief. PMID- 16382718 TI - [Laser treatment of BPH: evolution and perspectives in the French part of Switzerland]. PMID- 16382719 TI - [Prostatitis]. AB - Prostatitis, and particulary chronic pelvic pain syndrome, is a very challenging field in urology. Large randomized placebo treatment trials are necessary for a better evaluation of the different therapeutic options. This article is a review of the evolution in prostatitis diagnostic and actual treatments. PMID- 16382720 TI - [Benign prostatic hyperplasia and sexual dysfunction]. AB - Lower urinary tract symptoms of benign prostatic hyperplasia, erectile dysfunction and ejaculatory disorders are often associated. The severity of sexual dysfunction is positively associated with increasing severity of urinary tract symptoms. Current treatment of BHP produce some effects, positive as well as negative, on the sexual function. It is important to remember this, when you choose a treatment for a patient who report regular sexual activity. PMID- 16382721 TI - [Treatment of symptomatic prostatic hypertrophy with the KTP-80 laser]. AB - Macroscopic per and post-operative hematuria contributes significantly to hospitalisation time and to the morbidity of conventional surgery of symptomatic prostatic hypertrophy. It requires in fact transient anti-coagulation interruption, so as eventually a window with platelet anti-agregants. The newly available 80-Watts Potassium-Titanyl-Phosphate laser markedly reduces post operative hematuria, and allows operating the increasing population of patients under platelet anti-agregants, or event those who need therapeutic anticoagulation. This series reports on our initial experience with a technique which decreases remarkably post-operative catheterization and hospitalisation time, and whose obstruction relief is efficient, but needs to stand the test of time. PMID- 16382723 TI - [A boy or a girl: who decides?]. AB - In theory, humans produce an equal proportion of boys and girls. However, neither at conception nor at birth are both sexes equally represented. Environmental and individual changes are thought to play a role in the observed sex ratiodeviations. One of the key-mediator may be the individual (genetic) and environmental hormonal variations and their influence on gametogenesis, embryo implantation and development. PMID- 16382722 TI - [Hodgkin's disease]. PMID- 16382724 TI - [Medical opinion on right of opinion]. PMID- 16382725 TI - [The truth of Professor Hwang Woo-suk]. PMID- 16382726 TI - [The just indignation of Gheorghe Flutur, Romanian Agriculture Minister]. PMID- 16382727 TI - [French, Swiss and Norwegian researchers put forth new evidence of a new immune mechanism]. PMID- 16382728 TI - [Alzheimer's disease: a Genevian team on the free radical track]. PMID- 16382729 TI - [Economy and cheap junk]. PMID- 16382730 TI - Integrating integrative medicine--a how-to guide.. AB - Adding complementary and alternative medicine (CAM) to your hospital's treatment options doesn't have to be complicated or expensive--and the consumer appeal may be well worth it. PMID- 16382731 TI - Trustee workbook 4. Board self-evaluation turns 20: lessons learned and future trends. PMID- 16382732 TI - Strategic financial planning. What every trustee needs to know about facility replacement. AB - Find out what every trustee needs to know about facility replacement-when to rebuild, why and, most importantly, how to mesh new construction with a balanced financial and strategic plan. PMID- 16382733 TI - Good governance includes clinician participation. PMID- 16382734 TI - Managing CEO transition. PMID- 16382735 TI - Innovative hospital designs make a difference. AB - There is no better time for bringing health care delivery systems up to the cutting edge than when building a new hospital. Innovative designs can accommodate both patient and hospital staff needs while improving patient safety. PMID- 16382736 TI - Medicine and technology. PMID- 16382737 TI - Bulging tympanic membrane. PMID- 16382738 TI - Endoscopic view of 'mulberry hypertrophy' of the inferior turbinates. PMID- 16382739 TI - The paresis podule. PMID- 16382740 TI - Ethmoid fibrous dysplasia with anterior skull base and intraorbital extension. PMID- 16382741 TI - Vestibular findings in a 62-year-old woman with dizziness and a type I Chiari malformation. PMID- 16382742 TI - Eosinophilic esophagitis. PMID- 16382743 TI - Synergistic reconstruction of a cervicofacial defect using a free-tissue transfer and a prosthesis. PMID- 16382744 TI - Parathyroiditis. PMID- 16382745 TI - Office or operating room? PMID- 16382746 TI - A study of prognostic factors in sudden hearing loss. AB - We investigated the prognostic significance of the presence or absence of vertigo and tinnitus, the timing of the initiation of treatment, the type and severity of hearing loss, and age in 72 patients who had experienced sudden hearing loss. We found that the factors associated with a positive prognosis were the absence of vertigo, the presence of tinnitus, initiation of treatment within 7 days, a greater degree of hearing loss in the low frequencies, and a hearing loss of less than 45 dB. Age had no effect on prognosis. PMID- 16382747 TI - Nonsurgical home treatment of middle ear effusion and associated hearing loss in children. Part II: Validation study. AB - In this prospective follow-up investigation, we examined the efficacy of a modified Politzer device in the home treatment of persistent middle ear effusion (MEE) and associated hearing loss in children who had previously participated in a similar clinical trial. Our study group was made up of 38 patients who had been either (1) untreated control participants in the previous study whose hearing in one or both ears had not returned to normal within 11 weeks of their initial audiologic pretest ("former control group"; n = 30), or (2) active-treatment participants in the previous study whose hearing sensitivity in at least one ear had not improved to within normal limits after treatment and who elected to undergo a continuation of treatment ("extended-treatment group"; n = 8). Treatment efficacy was determined by comparing differences in pre- and posttreatment air-conduction thresholds and otoscopic findings. Following treatment, the former control group experienced significant improvements in hearing sensitivity at all frequencies; at the posttreatment test, hearing sensitivity was within normal limits in 43 of 60 ears (71.7%), and normal or moderate tympanic membrane mobility was observed in 30 of 34 otoscopically examined ears (88.2%). In the extended-treatment group, hearing sensitivity returned to within normal limits in 9 of 10 impaired ears (90.0%). These findings further substantiate the efficacy of our modified Politzer device in improving middle ear function and hearing sensitivity in children with MEE, and they support the reliability of the findings reported in our previous study. These results also indicate that many patients in whom initial treatment is not successful may benefit from extended treatment. PMID- 16382748 TI - Nasal glioma (neuroglial heterotopia) mimicking an astrocytoma: case report. AB - Nasal glioma is a rare benign tumor that usually occurs during infancy. We report a case of nasal glioma in a 6-month-old boy in which the histomorphologic features resembled those of an anaplastic astrocytoma. PMID- 16382749 TI - Surgical emphysema following tonsillectomy. AB - Complications of tonsillectomy have been well documented. However subcutaneous emphysema of the neck following tonsillectomy has rarely been described. We report a case of this complication in a young man who forcefully performed Valsalva's maneuver following a tonsillectomy. PMID- 16382750 TI - Adenoid cystic carcinoma of the salivary glands: a 20-year review with long-term follow-up. AB - The behavior of adenoid cystic carcinoma (ACC) of the salivary glands has been shown to be unpredictable in terms of local and distant spread and mortality. We retrospectively studied 35 operations in 34 patients who had had a pathologic diagnosis of ACC of the salivary glands and who had been treated over a 20-year period and followed for a minimum of 10 years. We analyzed the effect that different factors had on outcomes. The site of origin appeared to be an important factor in survival rates; survival among patients with tumors that had originated in the parotid gland was fairly good, while survival among those with tumors that originated in the minor salivary glands was significantly worse. TNM staging was another significant factor in survival. Other poor prognostic indicators were local spread, nodal positivity, distant metastasis, and local and regional recurrence. Radiation and chemotherapy did not appear to be beneficial for patients with advanced disease. We recommend radical surgery with complete resection for all patients with ACC of the salivary glands and a careful assessment of the neck in patients with minor salivary gland tumors. PMID- 16382751 TI - The cytokine activity of HMGB1--extracellular escape of the nuclear protein. AB - High mobility group box 1 (HMGB1), a mobile chromatin protein, passively leaks from necrotic cells and signals neighboring cells that tissue damage has occurred. Resting, non-activated inflammatory cells such as monocytes or macrophages contain HMGB1 in the nuclear compartment. When activated by lipopolysaccharide or inflammatory cytokines, they actively translocate the nuclear HMGB1 into the cytoplasm; HMGB1 is then exocytosed. At least one receptor for extracellular HMGB1 has been identified. HMGB1 acts as a mediator of systematic inflammation; it causes different cells to divide, migrate or elicit an immune response. Here, we give an abridged review of the cytokine activity of HMGB1, including its secretion mechanism, the putative signal transduction pathways, and its role in several inflammatory diseases. Finally, we cite a few examples in which therapeutic administration of HMGB1 antagonists rescued mice from lethal sepsis, arthritis and liver damage. The new findings of HMGB1 as a cytokine provide a better understanding of inflammatory diseases, establishing a clinically relevant therapeutic target that is significantly more efficient than other known cytokines. PMID- 16382752 TI - Human metapneumovirus and community-acquired pneumonia in children. AB - BACKGROUND: Human metapneumovirus (hMPV) was first recognized in the Netherlands in 2001 from nasopharyngeal aspirate samples and was associated with respiratory tract illness in the pediatric population. This was the first report of metapneumovirus infections in community-acquired pneumonia in Taiwan. METHODS: A total of 116 nasopharyngeal aspirate samples from patients with community acquired pneumonia was examined by reverse transcriptase polymerase chain reaction (RT-PCR). Other respiratory tract pathogens were also examined. The clinical characteristics and laboratory data were analyzed. RESULTS: Out of the 116 patients, potential causative agents were detected in 95 (81.9%) patients. A total of six human metapneumovirus RT-PCR positive samples was identified. All of these had evidence of coinfection with bacteria (3 Streptococcus pneumoniae, 2 Mycoplasma pneumoniae, 1 Chlamydia pneumoniae). Coinfection with other respiratory viruses was also observed in two cases (1 influenza A, 1 parainfluenza type 3). The age distribution was seven to 11 years except for one patient who was two years of age (Case 1). The most common clinical findings were fever (6/6, 100%), cough (6/6,100%), rhinorrhea (5/6, 83.3%), rales (5/6, 83.3%) and wheezing (1/6, 16.7%). Chest radiographs revealed four with lobar patches and two with interstitial infiltrations. The mean duration of hospital stay was 5.5 +/- 2.8 days. All patients made a complete recovery. CONCLUSIONS: hMPV was identified in 5.2% of patients with community-acquired pneumonia. Our data showed a high rate of coinfection with hMPV in community-acquired pneumonia. Human metapneumovirus infection, like other respiratory viruses, may predispose to secondary bacterial pneumonia. PMID- 16382753 TI - Comparison of directional coronary atherectomy-based intervention and stenting alone in ostial lesions of the left anterior descending artery. AB - BACKGROUND: Balloon angioplasty yields suboptimal results and increases the rate of restenosis in the ostium of the left anterior descending coronary artery (LAD). Several non-balloon devices have been used to improve the outcome of dilatation of such lesions. METHODS: This investigation retrospectively compared the results of a directional atherectomy (DCA)-based intervention and stenting alone on the ostial lesions of the LAD, and the effect on the left circumflex artery (LCX) ostium. Sixty-five patients with a successful angioplasty of LAD ostial lesions were studied. Patients were divided into 2 groups: group I (DCA, n = 34) and group II (stenting, n = 31). RESULTS: After intervention and at the 3 month follow-up, group I exhibited a larger minimal lumen diameter (MLD), smaller diameter stenosis, and a lower loss index than group II. At 6 months, group I showed a lower cumulative restenosis rate of 32% as compared to 48% for group II (p = 0.04). In group I, the MLD of the LCX ostium had not changed after DCA or at follow-up. In contrast, the MLD of the LCX was significantly reduced with stenting alone and during follow-up (p < 0.01). The strategy of optimal debulking plus stenting exhibited a restenosis rate of 9%, in contrast to 33% in the optimal debulking alone strategy (p = 0.05) after 3 months. CONCLUSIONS: During the intervention for LAD ostial lesions, the DCA-based strategy seemed to be superior compared to stenting alone both in terms of acute and late target lesion revascularization and in terms of the risk of plaque shifting. The best results however were obtained when optimal DCA was combined with stenting. PMID- 16382754 TI - Levels and value of soluble P-selectin following acute myocardial infarction: evaluating the link between soluble P-selectin levels and recruitment of circulating white blood cells and the marker for the rapid diagnosis of chest pain. AB - BACKGROUND: Platelet activation that results from coronary plaque rupture is important in the pathogenesis of acute myocardial infarction (AMI). Soluble p selectin (sP-selectin) is crucial in modulating leukocyte adhesion to both platelets and endothelial cells during inflammatory response and thrombus formation. We hypothesized that sP-selectin, an index of both platelet activation and acute inflammation, rapidly increases and modulates the recruitment of circulating white blood cells (WBC) in patients following AMI. METHODS: We conducted a prospective cohort study of 142 consecutive patients with ST-segment elevated AMI of onset < 12 h who were undergoing primary percutaneous coronary intervention. Blood samples for plasma levels of sP-selectin were obtained in the catheterization laboratory before coronary angiography was performed. The plasma levels of sP-selectin were also measured in 30 risk control subjects and 20 healthy control subjects. RESULTS: The plasma level of sP-selectin and the circulating WBC count were significantly higher in patients with AMI than in either the risk control or healthy subjects (all of p values < 0.0001). Additionally, repeated measures of ANOVA demonstrated that there were no significant differences in plasma levels of sP-selectin (p > 0.10) in three intervals from the start of chest pain to blood sample collection (< 180 min, > or = 180 < 360, and > or = 360 < 720) following AMI. Correlation analysis demonstrated that the increase in the plasma level of sP-selectin was significantly related to the circulating WBC count (r = 0.248, p = 0.003). CONCLUSIONS: sP-selectin was markedly elevated in an early phase of AMI. sP selectin may be involved in modulating the recruitment of circulating WBC during AMI. These findings raise the need for a prospective investigation of sP-selectin as a potential reliable clinical tool for rapidly diagnosing AMI. PMID- 16382755 TI - The effect of supplementation of docosahexaenoic acid and arachidonic acid on visual acuity and neurodevelopment in larger preterm infants. AB - BACKGROUND: Preterm infants may be born with deficits of both docosahexaenoic acid (DHA) and arachidonic acid (AA), but studies on supplementation of DHA and AA for preterm infants are limited. METHODS: Preterm infants with a gestational age between 30 and 37 weeks who met all the inclusion criteria were enrolled in this double blind, randomized, comparative study. Infants over 2000 g body weight, over 32 weeks of gestation and in full feeding status would enter into the active intervention period of 6 months. Sixteen infants received Neoangelac Plus with AA and DHA supplementation. Eleven infants received Neoangelac without AA and DHA supplementation. The babies had scheduled physical examinations and their cognitive development, visual acuity, and vital signs to be checked. Adverse events were also recorded. RESULTS: The mean Mental Development Index (MDI) scores for the supplementation and non-supplementation groups were 96.1 +/- 8.6 and 91.7 +/- 10.4 respectively at 6 months and 98.7 +/- 8.0 and 90.5 +/- 6.9 respectively at 1 year. The mean Physical Development Index (PDI) scores of these two groups were 102.2 +/- 10.5 and 95.4 +/- 13.2 respectively at 6 months and 98.0 +/- 5.8 and 86.7 +/- 11.1 respectively at 1 year. By repeated measures ANOVA, significant differences existed between groups for MDI and PDI (p = 0.020 and 0.008). However, there were no differences in visual acuity, physical examination variables or vital signs between these two groups. No obvious adverse effects were observed during the study period. CONCLUSION: These results showed possible benefits in the neurodevelopment of larger preterm infants given formula supplemented with DHA and AA. PMID- 16382756 TI - Leprosy in the Department of Dermatology, Chang Gung Memorial Hospital at Kaohsiung from 1988 to 2004: a clinical and histopathologic study of 13 cases. AB - BACKGROUND: Leprosy has long been in Taiwan, but it has never been eradicated. Incidental cases are easily overlooked nowadays because most younger dermatologists are unfamiliar with this disease. METHODS: We review and analyze 13 cases diagnosed as leprosy at the Department of Dermatology, Chang Gung Memorial Hospital at Kaohsiung from 1988 to 2004, all of which were histopathologically proven. RESULTS: The ages of the 13 recruited patients ranged from 31 to 73 (mean, 58.6) years, without a gender preference (male: female, 7:6). Two male patients were under 40 years old; one was a foreign worker from Thailand and the other was a local person in Penghu working as the chief officer on a fishing boat. The most-common clinical subtype was lepromatous leprosy (5/13), followed by borderline lepromatous leprosy, borderline tuberculoid leprosy, and tuberculoid leprosy (each 2/13), and then borderline leprosy and indeterminate leprosy (each 1/13). The initial clinical impression before the histopathological diagnosis included granuloma annulare, generalized eczema, lymphoma, syphilis, papular urticaria, cutaneous tuberculous infection, Sweet's syndrome, erythema annulare centrifugum, and hematoma. Most of these patients only received irregular treatment after the diagnosis was made and were soon lost to follow-up. CONCLUSIONS: With increasing numbers of foreign workers and immigrants living in Taiwan in recent years, leprosy seems to have become a challenging diagnosis for younger dermatologists. Dermatologists should keep this ancient disease in mind and not overlook it. Because of the necessity of prolonged medication, complete treatment and long-term follow-up of leprosy cases will remain a major problem in public health. PMID- 16382757 TI - Bronchoscopic electrocautery for palliation of post-anastomotic tracheal stricture in a patient with complete tracheal transection following blunt chest trauma. AB - There are many strategies available to treat palliation of airway obstruction due to benign or malignant conditions. The initial choice depends on the urgency of the situation, the extent of the disease process as assessed bronchoscopically, as well as the individual experience and preference of the physician. We present a rare case of complete tracheal transection following a traffic accident. Respiratory distress, which resulted from post-anastomotic tracheal stricture, developed progressively about 2 months after surgical repair. Symptomatic relief and improved ventilatory function were achieved in this patient once patency of the trachea was restored successfully using bronchoscopic electrocautery. The technique is a straightforward, safe, and quick method to palliate airway obstruction. PMID- 16382758 TI - The rehabilitation outcome of spinal meningioma induced proprioception deficit. AB - Chronic non-traumatic myelopathy developed in a woman who presented symptoms of gait ataxia, right leg motor weakness, dysesthesia and urinary difficulty. Clinical evaluation revealed right leg weakness and global anesthesia as well as temperature, vibratory and proprioception sensation loss below the T-9 level and deep tendon hyperreflexia over the lower extremities. Magnetic resonance imaging showed an extra-intramedullary meningioma at the T-9 level. Following microscopic subtotal excision of the tumor, the patient underwent satisfactory rehabilitation programs with outstanding outcomes presented using the neurological scoring system, functional balance grade, postural analysis of Baropodometry screen and single leg standing time. PMID- 16382759 TI - Diagnosis of nodal Langerhans cell histiocytosis by fine needle aspiration cytology. AB - Langerhans cell histiocytosis (LCH) is a rare disease and disease confined to the lymph nodes is even more uncommon. Fine needle aspiration (FNA) cytology of LCH of the lymph nodes has rarely been described. A case study of LCH of the lymph nodes in a 23-year-old man is presented. FNA smears showed high cellularity composed of many isolated Langerhans cells (LCs) with nuclear grooves admixed with numerous eosinophils, lymphocytes, giant cells, macrophages, and neutrophils. Further immunohistochemical study of the excised lymph node sections revealed that the histiocytes were positively stained with CD1a. The presence of LCs with nuclear grooves and eosinophils suggests the possibility of LCH. FNA cytology is a valuable method for diagnosis. PMID- 16382760 TI - Coronary revascularization in multivessel disease. Which is better, stents or surgery? AB - Coronary artery disease (CAD) remains the major cardiovascular health issue in contemporary clinical practice. Treatment options for multivessel CAD include medical therapy, percutaneous coronary intervention (PCI), and coronary artery bypass grafting (CABG). Here, the authors review the most recent data that compare intracoronary stenting to CABG in multivessel disease. They address therapeutic issues surrounding proper selection of the optimal revascularization strategy and give special consideration to high-risk populations, such as patients with diabetes. PMID- 16382761 TI - Commentary: a new era in epilepsy care. Guidelines address first steps in diagnosis and treatment. PMID- 16382762 TI - Management of epilepsy in adults. Diagnosis guidelines. AB - In this first of two articles on new epilepsy guidelines for primary care physicians, the authors present detailed algorithms for the diagnosis and classification of seizure disorders in adults. They discuss the differentiation between generalized and partial seizures and stress that accurate identification is especially important because the type of seizure determines the appropriate treatment. The second article (page 29) looks at the treatment portion of the new guidelines. PMID- 16382763 TI - Management of epilepsy in adults. Treatment guidelines. AB - In this second of two articles on new epilepsy guidelines for primary care physicians, the authors discuss which treatments are the most effective in adults with this disease. They describe the antiepileptic drugs that are currently approved and elucidate the differences between established and newer options. Detailed algorithms cover pharmacotherapy, follow-up, and ongoing maintenance. PMID- 16382764 TI - Using beta-blockers to cut perioperative risk in CAD. Cardioprotective strategies for noncardiac surgery. AB - Postoperative adverse myocardial ischemic events, such as infarction, unstable angina, and cardiac death, are common in patients with coronary artery disease (CAD). These events can be prevented in many patients with strategies such as the perioperative use of beta-blockers. In this article, the authors present four cases and discuss perioperative evaluation of patients having a noncardiac operation to determine risk of an adverse cardiac event after the procedure. Evidenced-based approaches to reducing the chances of an undesirable outcome in high-risk patients, such as preoperative testing and procedures and the use of beta-blockers, are presented. PMID- 16382765 TI - Koebner's phenomenon. Skin trauma may trigger psoriatic flares. PMID- 16382766 TI - Patient notes: Bipolar disorder. PMID- 16382767 TI - Environmental health: another advocacy issue for nurses. PMID- 16382768 TI - Joint statement on pain management. PMID- 16382770 TI - The sheer essence of nursing. PMID- 16382771 TI - Interesting HIPAA developments. PMID- 16382772 TI - Hurricane Katrina: stories from the field. PMID- 16382773 TI - Faculty receive million dollar NIH grants to continue their research. PMID- 16382774 TI - Profile of a specialty group--Florida Council of Perioperative Registered Nurses. PMID- 16382775 TI - Update on the Florida Center for Nursing. Statewide strategic plan for nursing workforce in Florida progress report as of October 2005. PMID- 16382776 TI - Illustration of dynamic changes in Alzheimer pathology: from mild cognitive impairment to terminal stage. AB - It is quite important to examine the neuropathology of large numbers of brains from non-demented as well as demented subjects to elucidate the pathogenesis of dementia. Such a study, "power neuropathology" reveals the temporal profile of the disease, and results in the development of the treatment and the prevention of dementia. PMID- 16382777 TI - Hereditary cerebral hemorrhage with amyloidosis-Dutch type. AB - The amyloid beta-protein (Abeta) E22Q mutation of the rare disorder hereditary cerebral hemorrhage with amyloidosis-Dutch type (HCHWA-D) causes severe cerebral amyloid angiopathy (CAA) with hemorrhagic strokes of mid-life onset and dementia. The mutation does not affect total Abeta production but may alter the Abeta1 42:Abeta1-40 ratio, and affect the proteolytic degradation of Abeta and its transport across the blood-brain barrier. Abeta E22Q aggregates faster into more stable amyloid-like fibrils than wild-type Abeta. Non-fibrillar Abeta(x-42) deposits precede the appearance of fibrils and the deposition of Abeta(x-40) in the vascular basement membrane. CAA severity tends to increase with age but may vary greatly among patients of comparable ages. Lumenal narrowing of affected blood vessels, leukoencephalopathy, CAA-associated vasculopathies, and perivascular astrocytosis, microgliosis, and neuritic degeneration complicate the development of HCHWA-D CAA. Parenchymal Abeta deposition is also enhanced in the HCHWA-D brain with non-fibrillar membrane-bound Abeta(x-42) deposits evolving into relatively fibrillar diffuse plaques variously associated with reactive astrocytes, activated microglia, and degenerating neurites. Plaque density tends to decrease with age. Neurofibrillary degeneration is absent or limited. HCHWA-D dementia is associated with CAA severity independently of Braak stage, age, and plaque density. Particularly, microaneurysms may contribute to the development of (small) hemorrhages/infarcts and the latter to cognitive decline in affected subjects. However, the relative importance of cerebral hemorrhages/infarcts, white matter damage and/or other CAA- or Abeta-related factors for cognitive deterioration in HCHWA-D remains to be determined. PMID- 16382778 TI - Spatial patterns of the pathological changes in the temporal lobe of patients with neuronal intermediate filament inclusion disease. AB - Neuronal intermediate filament inclusion disease (NIFID) is a new neurodegenerative disease characterized histologically by the presence of neuronal cytoplasmic inclusions (NI) immunopositive for intermediate filament proteins, neuronal loss, swollen achromatic neurons (SN), and gliosis. We studied the spatial patterns of these pathological changes parallel to the pia mater in gyri of the temporal lobe in four cases of NIFID. Both the NI and SN occurred in clusters that were regularly distributed parallel to the pia mater, the cluster sizes of the SN being significantly greater than those of the NI. In a significant proportion of areas studied, there was a spatial correlation between the clusters of NI and those of the SN and with the density of the surviving neurons. In addition, the clusters of surviving neurons were negatively correlated (out of phase) with the clusters of glial cell nuclei. The pattern of clustering of these histological features suggests that there is degeneration of the cortico-cortical projections in NIFID leading to the formation of NI and SN within the same vertical columns of cells. The glial cell reaction may be a response to the loss of neurons rather than to the appearance of the NI or SN. PMID- 16382779 TI - Widespread and abundant alpha-synuclein pathology in a neurologically unimpaired subject. AB - The intracytoplasmic aggregation of alpha-synuclein (alphaS) protein is a common denominator for a group of neurodegenerative disorders currently known as synucleinopathies. It is generally assumed that the incorporation of alphaS protein into compact inclusions compromises the function and viability of its host cell via mechanical disruption. Herein, we report a widespread and abundant alphaS pathology in an elderly subject, whose medical history gave no indication of any neurodegenerative disease. We compared neuronal and glial components in this neurologically unimpaired subject with a patient with a clinical syndrome of dementia with Lewy bodies (DLB) by using a range of antigenic determinants and an in situ end-labeling technique. We detected no differences in vascular pathologies, in gliosis, or in apoptosis that would have explained the incompatible clinical end-points. With respect to the Alzheimer's disease-related changes, the only differences noted were the beta-amyloid aggregates in the putamen found in the DLB patient alone. Our findings suggest that there must be some currently unidentified factors rather than alphaS-positive inclusions that are responsible for the neuronal dysfunction. The alphaS-positive inclusions may well represent detoxified reserves that cells can tolerate for years, and thus prevention of their development could actually accelerate the diseases process. PMID- 16382780 TI - Pocket proteins p107 and p130 exhibit increased expression in macrophages during SIV encephalitis. AB - Progression of HIV encephalitis (HIVE) is associated with neuronal damage and loss because of infiltration of infected and/or activated macrophages into the CNS. We have previously observed increased inactivation of the retinoblastoma susceptibility gene product (pRb) by phosphorylation in neurons and glia of HIVE and the simian model of HIVE (SIVE). To determine if other pRb family members are altered in response to increased macrophage-secreted factors, we investigated expression of pRb family members p107 and p130 in SIVE. Both p130 and p107 exhibited increased staining in macrophages, but not neurons, astrocytes or T cells in SIVE. Increased p130 and p107 immunostaining was not limited to virally infected or PCNA-expressing macrophages. Most p107-positive staining was observed in perivascular macrophages, suggesting p107 may indicate macrophages at a specific stage of differentiation soon after migration. In contrast, cytoplasmic p130 was found in the majority of macrophages present in SIVE cases and may indicate activation as it was not seen in microglia in control CNS. These findings suggest that p107 and p130 are differentially expressed in CNS macrophage populations which may have multiple derivations and/or roles in lentiviral encephalitis. PMID- 16382781 TI - "Forme fruste" of amyotrophic lateral sclerosis with dementia: a report of five autopsy cases without dementia and with ubiquitinated intraneuronal inclusions. AB - We clinicopathologically investigated five autopsy cases of ALS without dementia and with ubiquitinated intraneuronal inclusions. The age at onset of symptoms ranged from 52 to 81 years and the duration of the disease was from 10 months to 3 years, 3 months. All five patients initially developed lower motor neuron signs, including bulbar signs, and upper motor neuron signs were found in the middle to late clinical stages, but dementia was not observed in all five cases. Thus, the clinical diagnoses of all five patients were ALS without dementia. Neuropathological examination of all five cases revealed not only obvious degeneration of upper motor neurons with neuronal loss of Betz cells, but also lower motor neuron involvement associated with Bunina bodies. In addition, ubiquitin-immunoreactive intraneuronal inclusions in the hippocampal dentate granular cells and degeneration of the substantia nigra were observed in all five cases. Furthermore, neuronal loss with astrocytosis in the dorsomedial portion of the anterior first temporal gyrus was observed in all three cases in which this structure was examined. Neuronal loss with astrocytosis in the subiculum was found in four cases. Neuronal loss of the parahippocampal gyrus was observed in three of the five autopsy cases, and amygdala involvement was encountered in three of four cases in which this structure was investigated. Based on these clinicopathological findings and a review of the published literature, we concluded that our five cases were ALS without dementia, but with pathological hallmarks compatible with ALS with dementia. We also concluded that there is a "forme fruste" of ALS with dementia showing no overt dementia clinically. PMID- 16382782 TI - Pineal germinoma associated with multiple congenital melanocytic nevi: a unique presentation. AB - Intracranial germ cell tumors are rarely seen and typically localize in the pineal or suprasellar region. The largest category of germ cell tumors is the germinoma. There have been reported associations of malignant tumors and chromosomal abnormalities in germ cell tumors. In this study, we present a 22 year-old man with multiple congenital melanocytic nevi in association with pineal tumor. Congenital melanocytic lesions greater than 2 cm were counted to be 54 in number, and those smaller than 1 cm in diameter were found to be 25 in number. The pathological diagnosis of the pineal tumors was germinoma, and the lesions located in the occipital region and trunk were compound congenital nevi. To our knowledge, a relationship between multiple congenital melanocytic nevi and germ cell tumors has not been reported before. The connection between them remains to be clarified. PMID- 16382783 TI - Epithelioid malignant peripheral nerve sheath tumor involving maxillary sinus. AB - We present a case of epithelioid malignant peripheral nerve sheath tumor (MPNST) located in the maxillary sinus region in a young man. The clinical history was short, but at admission the neoplastic infiltration was so extensive that only diagnostic biopsy was performed. The patient received palliative treatment and died 6 months later. Histologically, the neoplasm had a predominant epithelioid component. Neoplastic tissue was vimentin, S-100, Cam 5.2 and neuron-specific enolase positive. P53 protein reaction was found in 6% of the cells and the proliferation index assessed with Ki-67 was 52%. An appropriate immunohistochemical panel was essential for the final diagnosis of this epithelioid malignant tumor, with the location rather unusual for MPNST. PMID- 16382784 TI - Autopsy case of hereditary spastic paraplegia with thin corpus callosum showing severe gliosis in the cerebral white matter. AB - We report an autopsy case of a 51-year-old man clinically diagnosed with a complicated type of hereditary spastic paraplegia. His sister showed similar manifestations. Gait disturbance was manifested at 14 years of age. Subsequently, slowly progressive spastic tetraplegia developed with mental deterioration, neuropathy and amyotrophy. Marked cerebral atrophy with thin corpus callosum was shown by cranial MRI. Autopsy revealed a severely atrophic brain with extreme thinning of the whole corpus callosum. Microscopically, neurodegeneration was found in the corticospinal tract, thalamus, cerebral white matter and substantia nigra, as well as in the anterior horn and posterior column of the spinal cord. The remaining neurons contained large amounts of lipofuscin and eosinophilic granules. Unique to this patient was the severe gliosis in the cerebral white matter and substantia nigra, suggesting that sufficient development had been established when the degenerative process occurred. The predominant feature of the present case is the neurodegeneration process rather than hypoplasia. PMID- 16382785 TI - Pathology of pure hippocampal sclerosis in a patient with dementia and Hodgkin's disease: the Ophelia syndrome. AB - An archive autopsy case of a 50-year-old man who died of Hodgkin's lymphoma had a 4-year, 4-month history of dementia. After radiochemotherapy, the lymphoma subsided except for involvement of the spleen, but the dementia remained. Neuropathological examination revealed that the pathology was confined to the hippocampus, both hippocampi showing sclerosis without inflammation. Neurons of sector cornu ammonis (CA) 1 were completely lost whereas moderate neuron loss was also observed in sectors CA3 and 4, and the dentate gyrus. Neurons of sector CA2 were relatively well preserved and the subiculum was intact. There was no evidence of global hypoxia, or of neurodegenerative disorders with pathological changes affecting the hippocampus. Although there was a long preneoplastic history, and no inflammatory changes were found at autopsy, the present case of hippocampal sclerosis could be included in the category of paraneoplastic limbic encephalitis associated with Hodgkin's lymphoma or the Ophelia syndrome. PMID- 16382786 TI - Disseminated intraparenchymal microgranulomas in the brainstem in central nervous system sarcoidosis. AB - We report a 70-year-old woman with sarcoidosis and multiple cranial nerve palsy. The patient suffered from dysarthria, dysphagia and weakness of the upper and lower extremities and died of sepsis. No abnormalities were noted in brain MRI. At autopsy, numerous epithelioid granulomas with Langhans giant cells were present in the bilateral lungs, including the hilar lymph nodes. The brain had a normal external appearance. Histologically, there were brainstem parenchymal lesions consisting of many microgranulomas, lymphocytic infiltration, activated microglias and astrocytosis. Perivascular lympocytic cuffing was also seen. Neither granulomas nor lymphocytic infiltration were seen in the leptomeninges. The present case was considered to be a peculiar type of neurosarcoidosis, that is, "sarcoid brainstem encephalitis". PMID- 16382787 TI - Development of a rat model of amyotrophic lateral sclerosis expressing a human SOD1 transgene. AB - Mutations in copper-zinc superoxide dismutase gene (SOD1) have been linked to some familial cases of ALS. We report here that rats that express a human SOD1 transgene with two different ALS-associated mutations (G93A and H46R) develop striking motor neuron degeneration and paralysis. By comparing the two transgenic rats with different SOD1 mutations, we demonstrate that the time course in these rats was similar to human SOD1-mediated familial ALS. As in the human disease and transgenic ALS mice, pathological analysis shows selective loss of motor neurons in the spinal cords of these transgenic rats. In addition, typical neuronal Lewy body-like hyaline inclusions as well as astrocytic hyaline inclusions identical to those in human familial ALS are observed in the spinal cords. The larger size of this rat model as compared with the ALS mice will facilitate studies involving manipulations of spinal fluid (implantation of intrathecal catheters for chronic therapeutic studies; CSF sampling) and spinal cord (e.g., direct administration of viral- and cell-mediated therapies). PMID- 16382788 TI - Peripheral nerve avulsion injuries as experimental models for adult motoneuron degeneration. AB - We have used adult rat peripheral nerve avulsion models to evaluate the effects of neuroprotective molecules on motoneuron degeneration. The right facial nerves of adult Fischer 344 male rats were avulsed and adenoviral vectors encoding glial cell line-derived neurotrophic factor (GDNF), brain-derived neurotrophic factor (BDNF), transforming growth factor-beta2 (TGFbeta2), and growth inhibitory factor (GIF) were injected into the facial canal. The treatment with the vectors significantly prevented the loss of lesioned facial motoneurons, improved choline acetyltransferase (ChAT) immunoreactivity and suppressed the induction of nitric oxide synthase activity in these neurons. In separate experiments, animals were orally administered a solution of a neuroprotective compound T-588 after avulsion. Both free oral administration and oral tube administration of T-588 improved the survival of injured motoneurons and ameliorated their ChAT immunoreactivity. These results indicate that the gene transfer of GDNF, BDNF, TGFbeta2, and GIF and oral administration of T-588 may prevent the degeneration of motoneurons in adult humans with motoneuron injury and motor neuron diseases. PMID- 16382789 TI - Is motoneuronal cell death in amyotrophic lateral sclerosis apoptosis? AB - To clarify the controversy concerning whether the cell death of motor neurons in ALS is apoptosis, we investigated the expression of Apaf-1 and caspase-9 mRNA in spinal cord tissue obained at autopsy from patients with ALS and controls using RT-PCR; the presence of in situ nuclear DNA fragmentation in motor neurons by the TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method; and immunocytochemical localization of Apaf-1 and caspase-3, which are known as promotors of apoptotic processes. Although Apaf-1 and caspase-9 mRNAs levels were increased in ALS, Apaf 1 immunoreactivity (IR) showed no significant difference between ALS and the control, and caspase-3 IR was not observed in ALS motoneurons, casting doubt on the notion that motor neurons in ALS undergo death by the classic apoptotic pathway. Although TUNEL-positive motor neurons were frequently observed in the anterior horn in ALS, these neurons always showed an atrophic cell body with a shrunken and pyknotic nucleus, indicating that they were at the terminal stage of degeneration. No apoptotic bodies were seen. These findings suggest that the mechanism of motor neuronal cell death in ALS might not be apoptosis, but some other as yet unidentified mechanism. PMID- 16382790 TI - Golgi apparatus of the motor neurons in patients with amyotrophic lateral sclerosis and in mice models of amyotrophic lateral sclerosis. AB - We examined the Golgi apparatus (GA) of motor neurons of patients with ALS and in mice models of ALS by immunohistological method using antiserum against MG160 and against components of the trans-Golgi network (TGN46). The GA of half of the remaining spinal cord motor neurons of patients with sporadic ALS showed fragmentation, where the GA were dispersed or fragmented into numerous small, isolated elements. The GA of Betz cells in sporadic ALS were fragmented similar to that of anterior horn cells, and the GA of spinal cord motor neurons of those with familial ALS and of those with ALS with basophilic inclusions were fragmented or diminished. The GA in the majority of the motor neurons contained Bunina bodies, basophilic inclusions and superoxide dismutase 1 (SOD1)-positive aggregates were fragmented. The motor neurons in transgenic mice expressing G93A mutation of the SOD1 gene showed the fragmentation of the GA months before the onset of paralysis. These findings suggest that the fragmentation of GA may be related to the neuronal degeneration in patients with ALS. PMID- 16382791 TI - A 29-year-old woman who presented with personality change and a tumor in the frontal skull base. PMID- 16382792 TI - Magnitude and pattern of neurological pathology in fatal dengue hemorrhagic fever: a summary of Thai cases. PMID- 16382793 TI - Research and teaching: developing strategies for a symbiotic relationship. PMID- 16382794 TI - From idea to graduation: the evolution of the first PhD program in a HBCU. AB - Hampton University has the distinction of being the first Historically Black College or University (HBCU) to fully implement a PhD in Nursing program. This was history making for our campus as the charter was changed to accommodate doctoral education. The PhD in Physics was the first doctoral program on campus, and Nursing was one of three doctoral programs that followed. Hampton University enrolled its first class of four students in the PhD in Nursing program in the fall of 1999. The major goal of the Hampton University PhD program in Nursing is the preparation of nurse scholars and researchers who can advance scientific knowledge and influence the development of effective health care policies and practices. This journey describes steps Hampton University has taken in addressing the demand for doctorally prepared nurses, especially minority nurses. Training more minority nurses impacts all communities, especially medically undeserved communities and minority populations. PMID- 16382795 TI - Coping among African-American daughters caring for aging parents. AB - BACKGROUND: A higher proportion of African-American caregivers reported having suffered physical and mental problems because of caregiving (U. S. Department of Health & Human Services, 2005). PURPOSE: The purpose of this study was to examine the coping strategies of African-American daughters who have functioned as caregivers. The Neuman Systems model was utilized as the framework for this study. METHOD: An exploratory design utilizing qualitative and quantitative methodologies was conducted in two phases. Phase I (N = 44) consisted of a series of focus groups sessions and Phase II (N = 106) participants completed the Basic Interview Schedule Survey. DISCUSSION: Findings revealed than other groups to report dementia and stroke in their care recipients that daughters who attended support groups had increased family involvement, were religious and coped better with caregiving. CONCLUSION: This study concluded that religion gave most participants a strong tolerance for the caregiving situation and served to mediate the caregiving strain. PMID- 16382796 TI - The influence of father absence on the self-esteem and self-reported sexual activity of rural southern adolescents. AB - The absence of the father in the resident home is an increasing trend in the United States, and the belief that the absence of fathers is lined with a variety of social pathologies is gaining national momentum. This study explored the relationship of father absence on self-esteem and self-reported sexual activity among rural southern adolescents. A sample of 1,409 adolescents (558 males and 851 females) aged 11 to 18 years was surveyed, and the Miller Self-Esteem Questionnaire (MSQ) was used to measure self-esteem. Analysis revealed that increased sexual activity occurred by adolescents in homes where no father present in the residence. Although a statistical significance was noted in the relationship between father absence and lower self-esteem, the magnitude of the difference was not large. Further, no relationship between self-esteem and sexual activity was noted. Seemingly, the absence of the father has a potentially detrimental effect on adolescents' lifestyle choices. Consideration of the notion that the phenomena of fathering rather than the mere presence of a father may contribute to differences in adolescent's lifestyle choices should be studied. PMID- 16382797 TI - Evaluation of Blackboard as a platform for distance education delivery. AB - The Hampton University School of Nursing has implemented a Pediatric Nurse Practitioner Distance Education Program that utilizes the Blackboard Learning System, which was designed to transform the Internet into a powerful environment for the educational experience. The Interne, however, may be a new learning environment for many nurses seeking a graduate degree. The Blackboard platform allows for synchronous, real-time activities such as chat rooms, and asynchronous, delayed, activities such as discussion boards and digital drop boxes that can be used for student-to-instructor and instructor-to-student document transfer. This article focuses on the skills and challenges of navigating a course on the Internet using Blackboard. Recommendations for improving the teaching-learning process using a Web-based environment are included. PMID- 16382798 TI - The magazine as mentor. PMID- 16382799 TI - Lights out? PMID- 16382800 TI - Phase compensation in single echo acquisition imaging. Phase effects of voxel sized coils in planar and cylindrical arrays. PMID- 16382801 TI - Integrated potentiostat for neurotransmitter sensing. A high sensitivity, wide range VLSI design and chip. PMID- 16382802 TI - Biocompatible parylene neurocages. Developing a robust method for live neural network studies. PMID- 16382803 TI - A biofidelic birthing simulator. Models for research and training in complicated human births. PMID- 16382804 TI - A new, easily miniaturized steerable endoscope. Squid tentacles provide inspiration for the Endo-Periscope. PMID- 16382805 TI - Recording brain activity wirelessly. Inductive powering in miniature implantable neural recording devices. PMID- 16382806 TI - A robotic vehicle for disabled children. Providing assisted mobility with the PALMA project. PMID- 16382807 TI - COMAR activities 2004-2005. PMID- 16382808 TI - Vagus nerve stimulation for the treatment of depression. PMID- 16382809 TI - The adventures of Sarah and Archie. PMID- 16382810 TI - How big are educational and racial fertility differentials in the U.S.? AB - Using pooled data from the 1980, 1985, 1990 and 1995 CPS and 1988 and 1995 NSFG surveys, we show that shifts in fertility timing have occurred disproportionately for the more educated and for whites (compared to the less educated and to African Americans). Such timing shifts imply that the underlying period quantum of fertility is considerably higher for college-educated women and for whites than suggested by the standard total fertility rate. Applying the Bongaarts Feeney model (1998), we decompose observed racial and educational differences in age-order-specific fertility rates and TFR into tempo and quantum components. We find that a modest part of educational differences and a substantial part of racial difference in period fertility can be attributed to differential changes in tempo. Analysis by race and education shows a clear interaction: higher fertility among African Americans is confined to the less educated. PMID- 16382812 TI - Ethnicity and fertility in Nigeria. AB - Using a sample of Hausa-Fulani, Yoruba, Ibo, and all other women from the 1990 Nigerian Demographic and Health Survey, this study examines ethnic fertility differentials in Nigeria within the context of the social characteristics and cultural hypotheses. Among all women, we find the net fertility of Hausa-Fulani women to be lower than that of Other women; with no statistically significant difference in the net fertility of Ibo, Yourba, and Other women. But, among currently married women, we find the net fertility of Hausa-Fulani and Yoruba women to be lower than that of Other women, while the net fertility of Ibo women is higher than that of Other women. Overall, the findings of this study are more consistent with the cultural hypothesis, because statistically significant fertility differentials by ethnicity remain, even after controlling for selected socioeconomic and demographic variables. PMID- 16382811 TI - Did births decline in the United States after the enactment of no-fault divorce law? AB - Previous research has demonstrated that U.S. no-fault divorce laws implemented between 1953 and 1987 resulted in more divorces in some states than would have occurred otherwise. In other states, divorce patterns appeared to follow prevailing trends even after implementation of no-fault divorce legislation. A more distal question is whether implementation of no-fault divorce laws had an effect on birth rates. We analyzed state-level birth data from all 50 states to assess the birth response to the enactment of no-fault divorce law in each state. Results suggested that birth rates decreased significantly two to four years following the enactment of no-fault divorce law for the group of 34 states whose divorce rates responded to no-fault divorce legislation. As predicted, among the 16 states whose divorce rates did not respond to no-fault divorce legislation, the enactment of no-fault divorce law had a small and nonsignificant positive influence on birth rates. Generally, the group of 34 states had lower post no fault birth rates than the group of 16 states. PMID- 16382813 TI - The influence of age at menarche on the fertility of Chinese women. AB - This paper examines the effect of age at menarche on children ever born (CEB). We use data from the 1997 Sample Survey of Population and Reproductive Health conducted by the China Population Information and Research Center and the State Family Planning Commission. Poisson regression models are estimated for 10,919 ever married Chinese Han women. The influence of a woman's age at menarche on her CEB is examined while controlling for the social effects of rural/urban residency, education, her number of fecund years, whether her first birth occurred before or after the initiation of China's one child policy, and her age at first marriage. The results support our hypothesized positive association between age at menarche and CEB. That is, the later a woman's age at menarche, the greater her number of children ever born. Holding the other five independent variables constant, we show that for every additional month in age at menarche, a Chinese Han woman's mean number of children ever born increases by 0.5 percent. Some of the implications of these results are explored. PMID- 16382814 TI - A divergent pattern of the sex difference in life expectancy: Sweden and Japan, early 1970s-late 1990s. AB - For most of the 20th century the sex gap in life expectancy in the industrialized countries has widened in favor of women. By the early 1980s a reversal in the long-term pattern of this differential had occurred in some countries, where it reached a maximum and thereafter followed a declining trend. Of particular interest to the present investigation is the anomalous experience of Japan, where unlike other high-income countries the female advantage in life expectancy has been expanding. We contrast the case of Japan with that of Sweden, where, like many other high-income nations, the sex differential in longevity has been narrowing in recent years. We observe that in Sweden, until the early 1980s, the sex gap in life expectancy (female-male) exceeded that of Japan; but this situation reversed in subsequent periods, when the Swedish differential narrowed and that of Japan widened. A decomposition analysis indicates that these divergent patterns since 1980 have resulted mainly from larger than expected reductions in male mortality in Sweden due to heart disease and from accidents and violence, lung cancer and "other" cancers. In Japan, death rates for men and women from heart disease--which is a leading cause of death--have tended to decline more or less at the same pace since the early 1980s; and with regard to lung cancer, and "other" neoplasms, male death rates in Japan have been rising while those of women have either declined or risen more slowly. Moreover, during the 1990s, male and female suicide rates rose in Japan, but the rates for men went up faster. Altogether, the net effect of these divergent mortality trends for men and women in Japan underlie much of the observed widening of its sex differential in longevity in recent years. PMID- 16382815 TI - Factors affecting nutritional status in female adults in Dhaka slums, Bangladesh. AB - This study looks at women from the slums in Mohammadpur, Dhaka, Bangladesh, where 54 percent of women's BMI was less than 18.5. Fifty percent of the Dhaka slum population lived below the poverty line. Logistic regression showed that women with income above 1,500 taka per capita were 1.78 times more likely to have a higher BMI (odds ratio 1.7863; CI = 0.671-3.639). Women with their own savings were 1.89 times more likely to have higher BMI (odds ratio 1.879; CI = 0.01163 1.6431). Women were 4.5 times more likely to have a higher BMI when food expenditure per capita above 559 taka per month (odds ratio 4.55; CI = 1.0302 8.0799). Women were 1.82 times more likely to have higher BMI when there was a break even situation in financial status (odds ratio 1.8212; CI = -015709 3.6285). Female headed households were 3.3 times more likely to have a higher BMI compared to women living in male headed households (odds ratio 3.2966; CI = 0.33711-6.25620). Women who work 15-23 days per month were 2.3 times more likely to have a higher BMI (odds ratio 2.33; CI = 0.1133-4.5600). Women who are the budget manager are 1.12 times more likely to have a higher BMI (odds ratio 1.125; CI = 0.29296-2.0966). Where as a husband who beats his wife is 1.83 more likely to have a poorer BMI (odds ratio 1.8312; CI = -3.72596-0.17508). Women who have no marriage documents and women who take days off due to illness less than 11 days per month were more likely to have a poorer BMI (odds ratio 0.5567; CI = 0.049339-2.8379; odds ratio 0.7569; CI = 0.183167-2.0002). Women's nutritional status and well being can influence their ability to provide for themselves and their families and the demonstration of a relationship between measures of women's autonomy and control in the household and women's nutritional status is an important indication of the importance of these sociological constructs. Women's participation in work outside the home may be a factor increasing their autonomy. The identification of relationships between women's autonomy and control and their physical well being should provide further leverage for policy change that will enable women to escape some traditional roles and to contribute as more equal partners with men in the future of Bangladeshi society. PMID- 16382816 TI - The effects of kin propinquity on infant mortality. AB - This study tests the grandmother hypothesis and analyzes the effect of kin propinquity on infant mortality in a 19th century American frontier communal, polygynous population. The study shows that the presence of maternal grandmothers, aunts, uncles, and paternal aunts were significantly associated with increased infant survivorship while grandfathers, paternal grandmothers, and paternal uncles showed little effect. This study has implications for understanding the evolution of a long postreproductive life span, postmarital residential strategies, and behavioral strategies that enhance inclusive fitness. PMID- 16382817 TI - A proposed feedback loop of sexually transmitted diseases and sexual behavior: the Red Queen's Dilemma. AB - (Micro)organisms, such as bacteria, which cause sexually transmitted diseases (STDs) in humans are presented with an interesting ecological challenge. These microorganisms need humans to have sexual contact with each other in order for the microorganisms to spread to other hosts as well as to have subsequent generations of descendants. However, diseases tend to lower the sex drive and to render the host less sexually attractive. It is argued that, over time, selective advantages sculpted organisms which cause STDs to be minimally symptomatic and to indirectly increase the number of sexual partners of the host. Neisseria gonorrhoeae which cause the STD gonorrhea are used as a prototype for these putative sexual dynamics. As a counter to the (micro)organisms' biological adaptations, human cultural innovations emerged and became integrated into the various traditions of social structures. PMID- 16382818 TI - Fracture pattern interpretation in the skull: differentiating blunt force from ballistics trauma using concentric fractures. AB - There have been several anthropological studies on trauma analysis in recent literature, but few studies have focused on the differences between the three mechanisms of trauma (sharp force trauma, blunt force trauma and ballistics trauma). The hypothesis of this study is that blunt force and ballistics fracture patterns in the skull can be differentiated using concentric fractures. Two hundred and eleven injuries from skulls exhibiting concentric fractures were examined to determine if the mechanism of trauma could be determined by beveling direction. Fractures occurring in buttressed and non-buttressed regions were examined separately. Contingency tables and Pearson's Chi-Square were used to evaluate the relationship between the two variables (the mechanism of trauma and the direction of beveling), while Pearson's r correlation was used to determine the strength of the relationship. Contingency tables and Chi-square tests among the entire sample, the buttressed areas, and the non-buttressed areas led to the null hypothesis (no relationship) to be rejected. Pearson's r correlation indicated that the relationship between the variables studied is greater than chance allocation. PMID- 16382819 TI - Semi-automated ultrasound facial soft tissue depth registration: method and validation. AB - A mobile and fast, semi-automatic ultrasound (US) system was developed for facial soft tissue depth registration. The system consists of an A-Scan ultrasound device connected to a portable PC with interfacing and controlling software. For 52 cephalometric landmarks, the system was tested for repeatability and accuracy by evaluating intra-observer agreement and comparing ultrasound and CT-scan results on 12 subjects planned for craniofacial surgery, respectively. A paired t test evaluating repeatability of the ultrasound measurements showed 5.7% (n = 3) of the landmarks being significantly different (p < 0.01). US and CT-scan results showed significant differences (p < 0.01) using a Wilcoxon signed rank test analysis for 11.5% (n = 6) of the landmarks. This is attributed to a difference in the volunteer's head position between lying (CT) and sitting (US). Based on these tests, we conclude that the proposed registration system and measurement protocol allows relatively fast (52 landmarks/20 min), non-invasive, repeatable and accurate acquisition of facial soft tissue depth measurements. PMID- 16382820 TI - The distal humerus--a blind test of Rogers' sexing technique using a documented skeletal collection. AB - Continuous monitoring of existing methods of skeletal diagnosis allows improving the reliability of personal identification in forensic and archaeological contexts. This study reports on a blind test re-evaluating the sexing technique proposed by Rogers (8) involving the distal humerus. A total of 351 humeri (184 male, 167 female specimens) from the documented skeletal assemblage of St. Bride's, London, was analyzed for the following traits: trochlear constriction, trochlear symmetry, olecranon fossa shape, and angle of the medial epicondyle. Individual traits showed substantial sex-discriminatory capacity, with "olecranon fossa shape" being most consistently accurate (84.6%) in predicting sex. The combination of all four traits provided an overall accuracy of 79.1%, including those individuals assessed as "probable" male and female. This renders the technique useful for forensic applications. The distal humerus can be recommended for sex assessment in addition to more established markers, especially since this part of the skeleton is frequently well preserved. PMID- 16382821 TI - Discriminating sex in South African blacks using patella dimensions. AB - For many years, sex determination has been carried out on skeletal remains to identify individuals in forensic cases and to assess populations in archaeological cases. Since it has been shown that not all bones are found in a forensic case, discriminant function equations should be derived for all bones of the body to assist in sex determination. Numerous studies have shown the usefulness of bones of the lower extremity (e.g. femur, tibia) in sex determination using discriminant function analysis, but the use of patella measurements has not been extensively investigated for this purpose. It is therefore the aim of this study to derive discriminant function equations for sex determination from measurements of the patella of South African blacks as represented in the Raymond A. Dart Collection of Human Skeletons. A total sample of 120 (60 male, 60 female) patellae were measured using six measurements. The Statistical Product and Service Solutions (SPSS) program was used to derive the equations. Stepwise and direct analyses were performed with the highest rate of classification of 85% thereby making the patella useful for sex determination. Thus, the proposed equations derived from this study should be used with caution and only on the South African black population group. PMID- 16382822 TI - An application of D- and L-aspartic acid mixtures as standard specimens for the chronological age estimation. AB - Instead of using the control teeth, we tried to prepare standard specimens to derive the equations for calculation of the age (analytical curves) with respect to each kind of tooth of various ages. To prepare standard specimens, we determined the racemization ratio of the teeth of known age that had actually been used for the appraisal of chronological age (total control teeth). Then we mixed commercially available L-Asp and D-Asp in the same D/L ratio as the measured one in the total control teeth. As a result, we were able to obtain the racemization rate equations from the age-specific standard specimens of central and lateral incisors. These equations were closely similar to those derived from actual teeth of known age. Since the racemization rate equations obtained from the standard specimens were satisfactorily reproducible, we assumed that these equations could be used in place of those obtained from the control teeth. Actually, in the age estimation of unidentified corpses from teeth, the use of standard specimens enabled us to estimate the age almost as precisely as estimated using the control teeth. Thus, the present study has demonstrated that in the estimation of chronological age the control teeth can be substituted by the standard specimens. This shows the possibility of using the standard specimens also in other laboratories where the racemization ratio can be measured with sufficient reproducibility. PMID- 16382823 TI - A test of the effectiveness of the revised maxillary suture obliteration method in estimating adult age at death. AB - The effectiveness of the original maxillary suture method for estimating age at death, introduced in 1987 by Mann and colleagues, has been tested, but their revised maxillary suture method (1991) has not been subject to similar scrutiny. The purpose of this study is to test the accuracy of the revised maxillary suture method in estimating age at death on a genetically diverse skeletal sample of 155 maxillae (96 males, 59 females, aged 26 to 100 years) of known age at death from the Department of Anatomy and Cell Biology, University of Cape Town, South Africa. Results from a prior study of the original maxillary suture method will be compared. With age category correctly estimated for 83% of this sample, the results of this study indicate that the revised method is more effective in estimating age at death than the original method. The revised method appears to perform best for older individuals and tends to underestimate age for individuals of all age groups. The results suggest that the revised method is useful as a method for age estimation when it is used conjunction with other estimators. PMID- 16382825 TI - Isotopic fractionation of carbon and nitrogen during the illicit processing of cocaine and heroin in South America. AB - The forensic application of stable isotope analysis to cocaine and heroin for geolocation of exhibits must take into account the possible enrichment and/or depletion of 13C and 15N during the illicit manufacturing process. Continuous flow elemental analysis-isotope ratio mass spectrometry was utilized to measure changes in the stable isotope ratios of carbon and nitrogen for both cocaine (N = 92) and heroin/morphine (N = 81) exhibits derived from illicit manufacturing processes utilized by South American clandestine chemists. In controlled settings in South America, there was no siginficiant carbon isotope fractionation during the conversion of cocaine base to cocaine HCI using current illict methodologies. In contrast, nitrogen isotope fractionation for this conversion was 1 per thousand. There was a kinetic carbon isotope ratio fractionation during the acetylation of Colombian morphine to heroin and as a result heroin exhibits will almost always have more negative delta13C values than the original morphine. There was an isotopic fractionation against 15N during the acetylation of morphine base to heroin base, but this effect was not expressed since all of the heroin base was precipitated during the manufacturing process. However, the clandestine process of converting a single batch of heroin base usually involved two consecutive crops of heroin HCl and the latter crop was isotopically depleted as expected from a Rayleigh distillation process. When heroin was deacetylated to morphine, the morphine produced resulted in delta13C values that were indistinguishable from the original morphine. The kinetic carbon isotope fractionation factor for the South American process of morphine acetylation was 1.8 per thousand, allowing calculation of the delta13C values of the acetic anhydride from deacetylated heroin delta13C values. PMID- 16382824 TI - Estimation of sex by 3D CT measurements of the foramen magnum. AB - The purpose of this prospective study is to investigate the value and accuracy of the measurements of the foramen magnum (FM) by using three-dimensional computed tomography (3DCT). Cases were randomly selected among 100 patients (48 males, 52 females) who had temporal CT in the Radiology Department. Seven measurements of the foramen magnum on 3D images, modified from the nine lines previously defined by Giles and Elliot were made. Using Fisher's linear discriminant functions test, the length and width of right condyle and width of FM diameters were found to be statistically different in each sex (p < 0.001) with 81% accuracy. To our best knowledge, this is the first report studying 3DCT measurements of FM, resulting with a sex determination accuracy rate of 81%. CT/3DCT can be reliably used in further investigations to provide basis for anthropometric and forensic issues. PMID- 16382826 TI - Informativeness of the CODIS STR loci for admixture analysis. AB - Population admixture (or ancestry) is used as an approach to gene discovery in complex diseases, particularly when the disease prevalence varies widely across geographic populations. Admixture analysis could be useful for forensics because an indication of a perpetrator's ancestry would narrow the pool of suspects for a particular crime. The purpose of this study was to use Fisher's information to identify informative sets of markers for admixture analysis. Using published founding population allele frequencies we test three marker sets for efficacy for estimating admixture: the FBI CODIS Core STR loci, the HGDP-CEPH Human Genome Diversity Cell Line Panel and the set of 39 ancestry informative SNPS from the Shriver lab at Pennsylvania State University. We conclude that the FBI CODIS Core STR set is valid for admixture analysis, but not the most precise. We recommend using a combination of the most informative markers from the HGDP-CEPH and Shriver loci sets. PMID- 16382827 TI - Development and evaluation of a standard method for the quantitative determination of elements in float glass samples by LA-ICP-MS. AB - Forensic analysis of glass samples was performed in different laboratories within the NITE-CRIME (Natural Isotopes and Trace Elements in Criminalistics and Environmental Forensics) European Network, using a variety of Laser Ablation Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS) systems. The main objective of the interlaboratory tests was to cross-validate the different combinations of laser ablation systems with different ICP-MS instruments. A first study using widely available samples, such as the NIST SRM 610 and NIST SRM 612 reference glasses, led to deviations in the determined concentrations for trace elements amongst the laboratories up to 60%. Extensive discussion among the laboratories and the production of new glass reference standards (FGS 1 and FGS 2) established an improved analytical protocol, which was tested on a well characterized float glass sample (FG 10-1 from the BKA Wiesbaden collection). Subsequently, interlaboratory tests produced improved results for nearly all elements with a deviation of < 10%, demonstrating that LA-ICP-MS can deliver absolute quantitative measurements on major, minor and trace elements in float glass samples for forensic and other purposes. PMID- 16382828 TI - Proton nuclear magnetic resonance spectroscopy (NMR) methods for determining the purity of reference drug standards and illicit forensic drug seizures. AB - A rapid, sensitive, accurate, precise, reproducible, and versatile method for determining the purity of reference drug standards and the routine analysis of illicit drugs and adulterants using proton (1H) Nuclear Magnetic Resonance (NMR) Spectroscopy is presented. The methodology uses a weighed sample dissolved in a deuterated solvent or solvent mixture containing a high purity internal standard. The NMR experiment employs 8 scans using a 45 second delay and 90 degrees pulse. In the determination of purity of reference standards, the number of quantitative determinations available is equal to the number of peak groups that are baseline resolved. The relative standard deviation (RSD) of these signals is usually < 1% for pure standards, and the results agree well with other purity determining methods. This method can also aid in the determination of correct molecular weight for standards containing an unknown number of waters of hydration or an unknown number of acids per drug in salts. Because the molar response for the hydrogen nucleus is 1 for all compounds, and since no separation media are used, only one linearity study is required to test a probe. In the presented study, the linearity of the NMR probe was determined using methamphetamine HCl dissolved in deuterium oxide (D2O) with maleic acid as the internal standard (5 mg) for a range of concentrations from 0.033 to 69.18 mg/ml with a resulting correlation coefficient of >0.9999 for all 6 methamphetamine peak groups. The spectra of complex illicit heroin, methamphetamine, MDMA, and cocaine samples are presented, as well as an extensive list of compounds, their solubilities and the solvent(s) and internal standard used. PMID- 16382829 TI - Empirical analysis of the STR profiles resulting from conceptual mixtures. AB - Samples containing DNA from two or more individuals can be difficult to interpret. Even ascertaining the number of contributors can be challenging and associated uncertainties can have dramatic effects on the interpretation of testing results. Using an FBI genotypes dataset, containing complete genotype information from the 13 Combined DNA Index System (CODIS) loci for 959 individuals, all possible mixtures of three individuals were exhaustively and empirically computed. Allele sharing between pairs of individuals in the original dataset, a randomized dataset and datasets of generated cousins and siblings was evaluated as were the number of loci that were necessary to reliably deduce the number of contributors present in simulated mixtures of four or less contributors. The relatively small number of alleles detectable at most CODIS loci and the fact that some alleles are likely to be shared between individuals within a population can make the maximum number of different alleles observed at any tested loci an unreliable indicator of the maximum number of contributors to a mixed DNA sample. This analysis does not use other data available from the electropherograms (such as peak height or peak area) to estimate the number of contributors to each mixture. As a result, the study represents a worst case analysis of mixture characterization. Within this dataset, approximately 3% of three-person mixtures would be mischaracterized as two-person mixtures and more than 70% of four-person mixtures would be mischaracterized as two- or three person mixtures using only the maximum number of alleles observed at any tested locus. PMID- 16382830 TI - Genipin, a novel fingerprint reagent with colorimetric and fluorogenic activity, part II: optimization, scope and limitations. AB - Genipin, a hydrolytic product of geniposide extracted from gardenia fruit, was thoroughly studied as a potential fingerprint reagent, and optimal conditions for fingerprint development have been determined. Latent fingerprints on paper items that have been treated with a non-ink running formulation containing 0.17% of the reagent, showed up as both colored and fluorescent images. On brown wrapping paper and on papers with highly luminescent backgrounds, genipin developed more visible and clearer prints than did classical reagents such as ninhydrin or DFO. Another potential advantage of genipin is that it is totally harmless and an environmentally friendly reagent. PMID- 16382831 TI - Stable isotope ratios as a tool in microbial forensics--part 3. Effect of culturing on agar-containing growth media. AB - Stable isotope ratios of hydrogen and oxygen in microbes have been shown to be functions of the corresponding isotope ratios of the water with which the culture medium was prepared, and thus to contain a potential geographic signal. Water can evaporate from agar (solid) media during culturing, changing its isotope ratios. Here we describe the effect of drying on the isotope ratios of water extracted from agar media and the H and O stable isotope ratios ratios of Bacillus subtilis spores cultured on agar. The delta2H vs delta18O relationship of water in Petri dish agar was surprisingly constant during evaporation regardless of the ambient relative humidity, making it possible to calculate the approximate isotope ratios of the original water, even in significantly evaporated agar. The H stable isotope ratios of spores cultured on agar remained relatively unchanged as the agar dried, but the O ratio became significantly enriched. PMID- 16382832 TI - The Microbial Rosetta Stone database: A common structure for microbial biosecurity threat agents. AB - Infectious microorganisms are important to multiple communities engaged in biodefense and biosecurity, including the agencies responsible for health, defense, law enforcement, agriculture, and drug and food safety. Many agencies have created lists of high priority infectious microorganisms to prioritize research efforts or to formally control the possession and distribution of specific organisms or toxins. However, the biological classification of infectious microorganisms is often complex and ambiguous, leading to uncertainty and confusion for scientists involved in biosecurity work. To address this problem, we created a database, known as the Microbial Rosetta Stone, which resolves many of these ambiguities and includes links to additional information on the microbes, such as gene sequence data and scientific literature. Here we discuss the efforts to coordinate organism names from pathogen lists from various governmental agencies according to biological relatedness and show the overlap of high-priority organisms from multiple agencies. To our knowledge, this is the first comprehensive coordination of pathogens, synonyms, and correct taxonomic names. The organized tables and visual aids are freely available at http://www.microbialrosettastone.com. This website provides a single location where access to information on a broad range of disease-causing organisms and toxins is available to members of the biosecurity community. PMID- 16382833 TI - Removing interfering contaminations from gelatin lifters. AB - Gelatin lifter is widely used for recovering shoeprints from crime scenes. Dusty shoeprints removed from paper with loose fibers, cloth or plasterboard, might be concealed by the detached fibers. A novel technique to clean the gelatin lifter from the interfering contaminations, using adhesive lifters, was developed. The adhesive lifter is applied directly on the surface of the gelatin lifter, and is removed instantly. The adhesive lifter removes the upper layer of the attached material on the gelatin lifter. After removing the concealing material from the hidden imprint, the quality of the visualized imprint is improved. The cleaning process can be applied as many times as needed and the optimum number is different for each substrate. The small dust particles comprising the shoeprint remained attached to the sticky side of the gelatin lifter even though the adhesion force applied by the adhesive lifter is great. Repeating the procedure too many times could harm the quality of the print severely, yet the advantages of this method are greater than the risks of relocation of image quality reduction. PMID- 16382834 TI - Fungal DNA challenge in human STR typing of bone samples. AB - The present study focuses on possible cross-reaction of fungal DNA with human STR primers that may affect subsequent forensic DNA analysis of forensic samples. Specificity of human STR markers namely HUMAMEL, HUMCSF1PO, D8S306, HUMTH01, HUMvWA, HUMFES/FPS, HUMF13A01, HUMDHFRP2, HUMFGA and HUMTPOX was tested using DNA of 24 different filamentous fungal isolates obtained from exhumed bone samples. The specificity of these ten STR markers for human DNA was demonstrated. Presence of non-human DNA in five bone samples analyzed did not alter scoring of detected alleles. Notably, amplification was inhibited in the presence of a high proportion of fungal DNA compared to human DNA (1000 ng: 1 ng) in DNA mixture experiments. The results of the present study underscore the importance of carefully analyzing the presence of non-human biological contaminants that may affect DNA typing of environmentally challenged forensic samples to avoid spurious data interpretation. PMID- 16382835 TI - Alkaloid content of the seeds from Erythroxylum Coca var. Coca. AB - Alkaloid extracts from the seeds of Erythroxylum Coca var. Coca grown in the Chapare Valley of Bolivia were subjected to gas and liquid chromatographic-mass spectrometric analyses. Several alkaloids from these seeds were detected and characterized, including methylecgonidine, tropine, 3alpha-acetoxytropane, ecgonine methyl ester, cuscohygrine, N-norbenzoyltropine, benzoyltropine, hexanoylecgonine methyl ester, cocaine, cis-cinnamoylcocaine, and trans cinnamoylcocaine. Methylecgonidine was determined to be the primary constituent and not an analytical artifact. Additionally, two significant new uncharacterized alkaloids were established as present. Recent evidence suggests that some cocaine processors are adding this seed extraction material to cocaine extracted from coca leaf and may impact cocaine impurity signature profiles. PMID- 16382836 TI - An ideal material for the preparation of known toolmark test impressions. AB - Traditionally, toolmark test exemplars are produced by applying a tool's working surface to a piece of soft metal such as lead. Soft, pliable metals are primarily used for this purpose because they will replicate the microscopic grooves present on a tool's working surface without damaging the tool. In this paper the authors present an alternative material for the preparation of test toolmarks. Jewelry modeling or carving waxes are utilized in this study. These waxes are designed for the jewelry modeling industry to create very fine, highly detailed wax models of jewelry pieces that will be cast in various metals utilizing the lost wax casting method. Jeweler's waxes have been found to be ideal for preparing test toolmarks from exemplar tools. The test tool's working surface is applied to a piece of the appropriate wax in a manner consistent with the tool's design. The replicas obtained are exact, highly detailed, 1:1, negative impressions of the exemplar tools working surface, have a long shelf-life, and are suitable for use in toolmark examination and comparison cases. PMID- 16382837 TI - Forensic value of pattern and particle transfers from deployed automotive airbag contact. AB - Many automobile manufacturers began installing airbags on the driver's side in the late 1980's. Passenger side airbags followed in the early 1990's. Most airbags use a solid-propellant type of material that produces a hot gas to inflate the airbags. The gas in the driver side airbag leaves the inflator at a temperature as hot as 600 degrees C. The hot gas escapes through the vent holes after deployment, but it can also leak through the stitching seams in the front and singe a pattern on the occupant's clothing characteristic of the seam pattern. The singe patterns from the driver and passenger side airbags will be different. Cornstarch, which is used as a lubricant in some driver side airbags, can transfer to the driver's shirt. Hairs, fibers and make-up can transfer from the driver or passenger to the surface of the deployed airbags. Two cases are presented, illustrating singe patterns and particle transfers, and how they helped determine who a driver or passenger were. PMID- 16382838 TI - Beware of the possibility of fingerprinting techniques transferring DNA. AB - Fingerprinting brushes have the potential to collect and transfer DNA during powdering. Squirrel-hair fingerprint brushes exposed to specific sets of saliva stains and brushes used in routine casework were tested for their ability to collect and transfer DNA containing material using standard DNA extraction procedures and AmpFlSTR Profiler Plus amplification and typing procedures. The tests found that the risk of transferring DNA during powdering and having a detrimental impact on the analysis increases if the examiner powders over either biological stains (such as blood or saliva) or very fresh prints and uses more sensitive PCR amplification and typing procedures. We advocate caution when powdering prints from which DNA may also be collected and provide options for consideration to limit the risk of transferred DNA contamination while fingerprinting. PMID- 16382839 TI - Drug smuggling using clothing impregnated with cocaine. AB - A case study is presented where a woman travelling from South America to the Republic of Ireland was detained at Dublin Airport and articles of clothing she had in her luggage were found to be impregnated with cocaine. The study shows that the amount of powder recovered from the garments was approximately 14% of the total weight of the garments. The cocaine was in the form of cocaine hydrochloride and the purity was approximately 80%. An examination of the garments under filtered light highlighted the areas exposed to cocaine and indicated that the method of impregnation was by pouring liquid containing cocaine onto the clothing. PMID- 16382840 TI - Skin penetration assessment of less lethal kinetic energy munitions. AB - The development of less-lethal technologies has provided law enforcement personnel with an alternative to lethal force. Although the less lethal projectile was produced to engender non-penetrating wounds, case studies show that there have been a number of reported penetrating injuries ranging from minor to significant in morbidity. The objective of this study was to determine the energy per unit area required to penetrate various regions of the body. Eight unembalmed postmortem human specimens were procured for this testing. Each specimen sustained a maximum of 25 impacts consisting of shots to the anterior and posterior thorax, abdomen, and legs. A 12-gauge, fin-stabilized, rubber rocket round was used as the impactor for all of the conducted tests. The energy density required for 50% risk of penetration varied from 23.99 J/cm2 for the location on the anterior rib (p = 0.000) to 52.74 J/cm2 for the location on the posterior rib (p = 0.001). PMID- 16382841 TI - Searching for the IRA "disappeared": ground-penetrating radar investigation of a churchyard burial site, Northern Ireland. AB - A search for the body of a victim of terrorist abduction and murder was made in a graveyard on the periphery of a major conurbation in Northern Ireland. The area is politically sensitive and the case of high profile. This required non invasive, completely non-destructive and rapid assessment of the scene. A MALA RAMAC ground-penetrating radar system was used to achieve these objectives. Unprocessed and processed 400 MHz data show the presence of a collapse feature above and around a known 1970s burial with no similar collapse above the suspect location. In the saturated, clay-rich sediments of the site, 200 MHz data offered no advantage over 400 MHz data. Unprocessed 100 MHz data shows a series of multiples in the known burial with no similar features in the suspect location. Processed 100 MHz lines defined the shape of the collapse around the known burial to 2 m depth, together with the geometry of the platform (1 m depth) the gravedigger used in the 1970s to construct the site. In addition, processed 100 MHz data showed both the dielectric contrast in and internal reflection geometry of the soil imported above the known grave. Thus the sequence, geometry, difference in infill and infill direction of the grave was reconstructed 30 years after burial. The suspect site showed no evidence of shallow or deep inhumation. Subsequently, the missing person's body was found some distance from this site, vindicating the results and interpretation from ground-penetrating radar. The acquisition, processing, collapse feature and sequence stratigraphic interpretation of the known burial and empty (suspect) burial site may be useful proxies for other, similar investigations. GPR was used to evaluate this site within 3 h of the survey commencing, using unprocessed data. An additional day of processing established that the suspect body did not reside here, which was counter to police and community intelligence. PMID- 16382842 TI - Seven hundred seventy eight bite marks: analysis by anatomic location, victim and biter demographics, type of crime, and legal disposition. AB - A study of the etiology, anatomic location, victim demographics and legal disposition of bite mark cases was made with the purpose of updating and augmenting previous research in the field. The information may be of interest to a myriad of professional disciplines including Forensic Odontologists, Medical Examiners, Detectives, Profilers, Emergency Room Personnel, Coroners, Psychologists, and Family Service Counselors, as bite marks provide both physical and biological data. While bite marks were found on all anatomic regions of the body some sites are significantly more likely to receive bites, and the frequency that an area is bitten may vary with the type of crime. Sex and age of the victim may also impact the resulting location and frequency of bites. A survey form for bite mark cases was created and mailed to all Diplomates of the American Board of Forensic Odontology. The survey form was also included in the American Society of Forensic Odontology newsletter. The survey requested that the recipient fill out a separate form for each case for which the recipient was the primary investigator of a patterned injury. The data from the resulting surveys were entered into a Microsoft Excel spreadsheet. The responses detailed two hundred thirty two (259) bite mark cases that included seven hundred (778) individual bite marks. Harvey (1976) and Sweet and Pretty (2000) published studies finding the highest percentage of bites to the breasts. In 1983 Vale and Noguchi published the paper indicating that the most frequently bitten area was the upper extremities. The survey forms were sent to approximately 1100 forensic dentist in 26 countries. The forensic experience level of the dentists varied from neophyte to very experienced. The data were analyzed and the results reported and organized in the following categories; Victim Distribution by Gender, Victim Distribution by Age, Child Abuse Distribution by Age and Gender, Sexual Assault Distribution by age and Gender, Homicide Distribution by Age and Gender, Bite Mark Distribution by Gender and Location, Number of Bite Marks per Victim, Bite mark Distribution Comparison to Previous Research, Child Abuse Suspect Age Distribution by Age and Sex, Homicide Suspect Age Distribution by Age and Sex, Sexual Crimes Suspect Age Distribution by Age and Sex, and Bite Mark Incidence by Anatomical Area and Type of Crime. Fifty-two forensic odontologists from seven countries responded. Nineteen responders were Diplomates of the American Board of Forensic Odontology. The number of cases reported by each responder ranged from one to thirty-three and the average number of cases reported was 4.5. In this broad based study, females were bitten more often than males. The average male victim was younger than the average female victim. Males that were victims tended to be either very young or very old. The youngest victim was a two-month-old boy and the oldest victim a 95-year-old woman. Perpetrators were male more often than female and there was an average of 1.4 suspects per case. The results show that most bites occurred on the arm, followed by the breast. If broken down by gender, males were bitten on the arm more than females, and females were bitten on the breast more often than males. The data show patterns in location and number of bites that seem related to both the type of crime and the age of the victim. PMID- 16382843 TI - Adolescent death: a 15-year retrospective review. AB - Adolescents comprise an eclectic mix of people vitally important to society yet long-term comprehensive studies on the circumstances of their deaths are lacking in the pediatric forensic literature. The authors reviewed all forensic cases referred to the Medical University of South Carolina Forensic Pathology section over the fifteen years between January 1989 and December 2003. In accordance with the World Health Organization (WHO) definition of adolescents, only decedents 10 19 years of age were included. The authors examined the cause and manner of death, age, gender, and race of the victims in all cases. The toxicology results, perpetrator, death scenario and location, and victim traits were analyzed when available. For all adolescents, the most common manner of death was accident followed by homicide, suicide, natural, and undetermined. Within the adolescent population two distinct groups, 10-14 years old and 15-19 years old, were identified. Though both groups were similar in that they experienced a high number of accidental deaths, decedents of older age group suffered a higher percentage of violent deaths while decedents of the younger group were more likely to die of natural causes. Many of the accidental deaths in this review were preventable, including deaths due to motor vehicle collisions and drowning. In deaths due to homicide, the perpetrator was often known to the victim, whether as an acquaintance or family member. Toxicology testing was often positive in decedents of the older age group, while only rarely positive in decedents of the younger age group. With a solid understanding of the circumstances, it may be possible to predict, and hopefully prevent, future cases of adolescent death. The authors present their findings in this 15-year retrospective study to better aid forensic pathologists, death investigators, law enforcement, and epidemiologists. PMID- 16382844 TI - Postmortem monocular indirect ophthalmoscopy. AB - Postmortem monocular indirect ophthalmoscopy permits examination of the posterior fundus and peripheral retina even if there is less than perfect anterior segment media such as postmortem corneal clouding. Light directed through the decedent's pupil from a bright focal light source illuminates the fundus and reflected light from the retina is then projected out of the eye. An aspheric condensing lens positioned in front of the eye focuses the retinal image at the focal plane of the lens. The real inverted, laterally reversed image comprises a wide field of view permitting evaluation of the decedent's fundus for retinal hemorrhages and other lesions. PMID- 16382845 TI - Diagnosis of Alzheimer's disease in an exhumed decomposed brain after twenty months of burial in a deep grave. AB - After 20 months of interment in a deep grave, the decomposed body of the 81-year old testator of a will was exhumed to sustain the burden of proof that he lacked testamentary capacity when the will was rewritten two days prior to his death. The brain was mushy and pulverized with complete disappearance of the brainstem, cerebellum and subcortical ganglia. Small foci of relatively intact dorsal frontal neocortex were identified. Sections from these foci were stained with hematoxylin and eosin, bielchowsky silver stain and immunostains for beta amyloid peptide (betaA4), tau and alpha-synuclein. Despite severe autolysis and decomposition, the bielchowsky stain and the betaA4 immunostains showed preserved frequent neuritic amyloid plaques with very few residual neurofibrillary tangles. Cerebral Amyloid Angiopathy was present. At the present time this case represents the first documented and reported case of direct tissue diagnosis of Alzheimer's Disease pathology in a decomposed brain following long term burial in a deep grave. PMID- 16382846 TI - The stalking of public figures: management and intervention. AB - Public figures are at relatively high risk of unusual contact initiated by fixated individuals. Prior research on managing the threat presented by public figure stalkers concludes that although direct threats rarely precede attacks, there is usually evidence of pre-planning. Furthermore, some public figure attackers do attempt to communicate with their future intended victim prior to attack. Thus, early warning signs from unusual contact behavior can be a powerful tool in threat assessment and risk management. The current paper offers a systematic concept for managing public figure stalking and constitutes five stages: (i) screening, (ii) first analysis, (iii) passive research, (iv) active research and finally (v) considered management strategy. It is concluded that assessment and management of risk are dynamic procedures, requiring ongoing monitoring and flexibility. Furthermore, although different stalkers will engage in ostensibly similar behavior, their motives and underlying psychopathologies may vary considerably. As such, all interventions require individual construction. PMID- 16382847 TI - Child murder committed by severely mentally III mothers: an examination of mothers found not guilty by reason of insanity. 2005 Honorable Mention/Richard Rosner Award for the best paper by a fellow in forensic psychiatry or forensic psychology. AB - Forensic hospital records of 39 severely mentally ill mothers adjudicated Not Guilty by Reason of Insanity for filicide (child murder by parents) were analyzed to describe characteristics preceding this tragedy and to suggest prevention strategies. Almost three-quarters of the mothers (72%) had previous mental health treatment. Over two thirds (69%) of the mothers were experiencing auditory hallucinations, most frequently command hallucinations, and half (49%) were depressed at the time of the offense. Over one third (38%) of the filicides occurred during pregnancy or the postpartum period, and many had a history of postpartum psychosis. Almost three-quarters (72%) of the mothers had experienced considerable developmental stressors, such as death of their own mother or incest. Maternal motives for filicide were predominantly "altruistic" (meaning murder out of love) or "acutely psychotic" (occurring in the throes of psychosis, without rational motive). Psychiatrists should perform careful risk assessments for filicide in mothers with mental illnesses. PMID- 16382848 TI - Some thoughts on the neurobiology of stalking. AB - The authors examine the crime of stalking, including the cognitive traits, emotional reactions, attachment pathology, violence patterns and sex differences of samples of stalking offenders. They focus on two common types of stalkers: 1) those who sustain pursuit of a former sexual intimate who has rejected them; and 2) those who pursue a stranger or acquaintance who has failed to return the stalker's romantic overtures. The authors discuss data from neuroimaging (fMRI) studies of romantic love which suggest that these forms of stalking may be associated with heightened activity of subcortical dopaminergic pathways of the "Reward System" of the brain, perhaps in combination with low activity of central serotonin. The authors propose that this set of neural correlates may contribute to the stalker's focused attention, increased energy, following behaviors, obsessive thinking about and impulsivity directed toward the victim. To further explore the neural systems associated with stalking behavior, they also discuss several biopsychological phenomena associated with romantic rejection, including the "protest response," "frustration attraction," "abandonment rage" and "mate guarding." They illustrate the parallels between stalking and addiction. They conclude that stalking may be associated with a specific set of biological components and they offer suggestions for further research into this pathological emotional/motivational state. PMID- 16382849 TI - Identification of anhydroecgonine ethyl ester in the urine of a drug overdose victim. AB - Toxicological evaluation of postmortem urine collected from a 41-year-old deceased white male detected anhydroecgonine ethyl ester (ethylecgonidine, AEEE), a transesterification product of smoked cocaine co-abused with ethanol. A solid phase extraction (SPE) method was used to extract cocaine, AEEE, and related metabolites from urine. SPE on a 1 mL urine sample from the decedent followed by GC-MS detected AEEE. Other metabolites identified by GC-MS included cocaine, cocaethylene, and anhydroecgonine methyl ester (AEME). To determine whether some or all of the AEEE was artifactually produced in the heated GC injector port, an alternative LC-MS method was developed. LC/MS following SPE found at least 50 ng/mL of AEEE in the extract. The mass fragmentation (MS/MS and MS3) of AEEE detected in the urine was compared to spectra of authentic, synthesized compound. AEEE is a potential additional forensic marker for the co-abuse of smoked cocaine and ethanol. PMID- 16382850 TI - Death of an infant involving benzocaine. AB - This report describes the death of a four-month-old Hispanic male which may be related to benzocaine toxicity. A toxicological evaluation revealed benzocaine at a concentration of 3.48 mg/L, and postmortem methemoglobin of 36% (normal 0.4 1.5). Methemoglobinemia is a complication of benzocaine toxicity. In light of the toxicology findings, the coroner investigated the source of the benzocaine and discovered that the child was treated with Zenith Goldline Allergen Ear Drops containing 0.25% w/v benzocaine and 5.4% w/v antipyrine. There was an admission by a caregiver that on the day prior to the child's death, he had been treated with three times the prescribed dose. Blood benzocaine concentrations in nine other unrelated cases were determined and concentrations ranged from <0.05-5.3 mg/L (mean 1.48 mg/L). Seven of the nine cases were positive for drugs of abuse, and one additional case was described as a known drug user. Methemoglobin in these benzocaine positive cases ranged from 6-69%; however, methemoglobin concentrations in postmortem cases are frequently elevated and should be interpreted with caution. The unknown significance of the benzocaine, and the circumstances of the case raise questions about the ultimate attribution of this death to SIDS. PMID- 16382851 TI - A hemizygous repeat polymorphism of one STR in four population groups of South India. PMID- 16382852 TI - Allele frequencies for two STR loci GATA31 E08 and DXS6809 in Chinese population. PMID- 16382853 TI - Genetic characteristics of three new Y-STRs: DYS631, DYS634 and DYS635 in a Chinese population. PMID- 16382854 TI - Autosomal STR frequencies in Afghanistan population. PMID- 16382855 TI - Unlinked tetrameric microsatellites on the X chromosome: frequency data in males from Cantabria (northern Spain). PMID- 16382856 TI - Genetic diversity of 15 STRs in Choles from northeast of Chiapas (Mexico). PMID- 16382857 TI - Y-STRs haplotypes of Han population in Xi'an of China. PMID- 16382858 TI - Allele frequencies for the 15 short tandem repeat loci in Slovenian population. PMID- 16382859 TI - Genetic polymorphisms of 15 STR loci in Chinese Hui population. PMID- 16382860 TI - Population data of 15 STR in Chinese Han population from north of Guangdong. PMID- 16382861 TI - Population data for the D5S818, D 13S317, D7S820 and D16S539 STR loci in a Romanian population sample. PMID- 16382862 TI - Allele frequency distribution of five X-chromosomal STR loci in an antioquian population sample (Colombia). PMID- 16382863 TI - Allele frequencies and statistic parameters for penta D and penta E loci in Chinese Han population. PMID- 16382864 TI - Haplotype distribution of the multi-copy Y-STR loci DYS385, DYS459 and DYS464 in Chinese Han population. PMID- 16382865 TI - Allele frequencies of 18 STR loci in Chinese population. PMID- 16382866 TI - Haplotype frequencies of four Y-chromosome STR loci in Chinese population. PMID- 16382867 TI - Commentary on: Budowle B, Shea B, Niezgoda S, Chakraborty R. CODIS STR loci data from 41 sample populations. J Forensic Sci 2001;46:453-489. PMID- 16382868 TI - Identification and analysis of microRNAs. PMID- 16382869 TI - Dormancy and the cell cycle. PMID- 16382870 TI - The OPT family functions in long-distance peptide and metal transport in plants. AB - The OPT family was first described six years ago, and much progress has been made in understanding the role these transporters play in their respective organisms. Plants are the only organisms in which both YS- and PT-type transporters have been characterized, and all of these OPTs appear to be plasma membrane-bound proteins, suggesting that they import substrates from the apoplasm or the environment. YS1 is the only OPT known to translocate substrates from the rhizosphere, whereas all the other OPTs seem to function in long-distance transport of peptides or metals. The sum of all the studies covered in this review suggest the model for OPT function in plants depicted in Figure 4. Peptides, metal-NA, and metal-MAs complexes (Strategy II plants only) are loaded into the xylem stream in the root for long-distance transport. OPTs unload the xylem by importing substrates into sink tissues such as leaves and by transloading the phloem. Peptides and metal-NA complexes exit the leaf symplasmically or by importation into the phloem from the apoplasm by OPTs. The filial tissues (endosperm and embryo) are apoplasmically separated from the maternal tissues, and OPTs may also function in loading the developing seed. Similarly, seedlings are symplasmically disconnected from the endosperm and OPTs may help move nutrients to the growing plant. Much progress has been made in the last two years toward understanding OPTs in plants, although several fundamental questions remain unanswered. Namely, what is the level of redundancy? Is there any substrate overlap between YS and PT OPTs? How crucial are their respective roles? Are there additional functions beyond peptide and metal transport? Given the recent pace of discovery, we may not have to wait long to find out the answers. PMID- 16382871 TI - Phospholipid-derived signaling in plant response to temperature and water stresses. PMID- 16382872 TI - Anionic nutrient transport in plants: the molecular basis of the sulfate transporter gene family. PMID- 16382873 TI - Purification of protein complexes by immunoaffinity chromatography: application to transcription machinery. PMID- 16382874 TI - Biogenesis of iron-sulfur cluster proteins in plastids. AB - Iron-sulfur (Fe-S) clusters are co-factors of proteins that perform a number of biological roles, including electron transfer, redox and non-redox catalysis, regulation of gene expression, and as sensors within all living organisms, prokaryotes and eukaryotes. These clusters are thought to be among the oldest structures found in biological cells. In chloroplasts, Fe-S clusters play a key role in photosynthetic electron transport as well as nitrogen and sulfur assimilation. The capacity of the Fe atom in Fe-S clusters to take up an electron reversibly provides the required electron carrier capacity in these pathways. Iron and sulfur limitation both affect plant primary production and growth. It has long been known that iron deficiency leads to defects in photosynthesis and bleaching in young leaves, phenomena that are closely linked to a defect in chloroplastic photosystem-I (PSI) accumulation, a major Fe-S containing protein complex in plants. Although the functional importance of Fe-S cluster proteins is evident and isolated chloroplasts have been shown to be able to synthesize their own Fe-S clusters, much is yet to be learned about the biosynthesis of Fe-S proteins in plastids. The recent discovery of a NifS-like protein in plastids has hinted to the existence of an assembly machinery related to bacterial Fe-S assembly systems. This chapter aims to summarize what we presently know about the assembly of Fe-S clusters in plants with an emphasis on green plastids. PMID- 16382875 TI - Iron transport and metabolism in plants. PMID- 16382876 TI - Salt stress signaling and mechanisms of plant salt tolerance. PMID- 16382877 TI - Strategies for high-throughput gene cloning and expression. AB - High-throughput approaches for gene cloning and expression require the development of new, nonstandard tools for use by molecular biologists and biochemists. We have developed and implemented a series of methods that enable the production of expression constructs in 96-well plate format. A screening process is described that facilitates the identification of bacterial clones expressing soluble protein. Application of the solubility screen then provides a plate map that identifies the location of wells containing clones producing soluble proteins. A series of semi-automated methods can then be applied for validation of solubility and production of freezer stocks for the protein production group. This process provides an 80% success rate for the identification of clones producing soluble protein and results in a significant decrease in the level of effort required for the labor-intensive components of validation and preparation of freezer stocks. This process is customized for large-scale structural genomics programs that rely on the production of large amounts of soluble proteins for crystallization trials. PMID- 16382878 TI - Molecular roles of chaperones in assisted folding and assembly of proteins. PMID- 16382879 TI - Engineering plants for increased nutrition and antioxidant content through the manipulation of the vitamin E pathway. PMID- 16382880 TI - Cancer stem cells--normal stem cells "Jedi" that went over to the "dark side". AB - Evidence has accumulated that cancer develops from a population of quiescent tissue committed/pluripotent stem cells (TCSC/PSC) or cells developmentally closely related to them that are distributed in various organs. To support this notion, stem cells (SC) are long lived cells and thus may become the subject of accumulating mutations that are crucial for initiation/progression of cancer. More important, they may maintain these mutations and pass them to the daughter stem cells. Therefore, mutations that occur in normal SC, accumulate during the life of an organism at the clonal level in the stem cell compartment committed to a given tissue/organ. As a consequence, this may lead to the malignant transformation of SC and tumor initiation. Furthermore, many biological features of normal and cancer SC such as the physiological trafficking of normal and metastasis of cancer stem cells involve similar molecular mechanisms, and we discuss these similarities here. Therefore, looking both at the origin and behavioral aspects we can envision cancer SC being normal SC "Jedi" that went over to the "dark side". PMID- 16382881 TI - Leukemic stem cells show the way. AB - The blood-related cancer leukemia was the first disease where human cancer stem cells (CSCs), or leukemic stem cells (LSCs), were isolated. The hematopoietic system is one of the best tissues for investigating cancer stem cells, since the developmental hierarchy of normal blood formation is well defined. Leukemia can now be viewed as aberrant hematopoietic processes initiated by rare leukemic stem cells (LSC) that have maintained or reacquired the capacity for indefinite proliferation through accumulated mutations and/or epigenetic changes. Yet, despite their critical importance, much remains to be learned about the developmental origin of LSC and the mechanisms responsible for their emergence in the course of the disease. This report will review our current knowledge on leukemic stem cell development and finally demonstrate how these discoveries provide a paradigm for identification of Cancer Stem Cell (CSC) from solid tumors. PMID- 16382882 TI - The marrow cell continuum: stochastic determinism. AB - Traditional models of hematopoiesis have been hierarchical in nature. Over the past 10 years, we have developed data indicating that hematopoiesis is regulated in a continuum with deterministic and stochastic components. We have shown that the most primitive stem cells, as represented by lineage negative rhodamine(low) Hoechst(low) murine marrow cells are continuously or intermittently cycling as determined by in vivo BrdU labeling. When marrow stem cells are induced to transit cell cycle by in vitro exposure to cytokines, either IL-3, IL-6, IL-11, and steel factor or thrombopoietin, FLT3 ligand, and steel factor, they progress through cycle in a highly synchronized fashion. We have determined that when the stem cells progress through a cytokine stimulated cell cycle the homing, engraftment, adhesion protein, global gene expression, and hematopoietic differentiation phenotypes all change in a reversible fashion. This has led to the continuum model, in which, with cycle transit, chromatin is continually changing altering open transcription areas and providing a continually changing landscape of transcriptional opportunity. More recently, we have extended the changing differentiation profiles to differentiation into lung cells and found that non-hematopoietic differentiation also shows cycle related reversibly modulation. These observations all together support a continuum model of stem cell regulation in which the phenotype of the marrow stem cells is continually and reversibly changing over time. PMID- 16382883 TI - Critical early events in hematopoietic cell seeding and engraftment. AB - Durable hematopoietic stem cell engraftment requires efficient homing to and seeding in the recipient bone marrow. Dissection of cellular and molecular mechanisms by retrospective analysis of functional engraftment studies imposes severe limitations on the understanding of the early stages of this process. We have established an experimental approach for in vivo functional imaging of labeled cells at the level of recipient bone marrow in real time. The adhesive interaction of hematopoietic cells with the bone marrow stroma evolves as the most important early event. Adhesion to the marrow, rather than the vascular endothelium, determines the efficiency of both homing and seeding, and is absolutely essential to maintain cell viability in the marrow. Seeding and engraftment may be improved either by bypassing homing or by localized transplant of a large number of cells in a relatively small marrow space. There is functional redundancy in the molecular pathways that mediate the cell-stroma interaction, such that blockage of a single pathway has only minor effect on homing and seeding. We hypothesize that successfully seeding-engrafting cells undergo extensive phenotypic changes as a consequence of interaction with the stroma, without engaging in rapid proliferation. Surprisingly, Fas-ligand appears to promote hematopoietic cell engraftment by immunomodulatory and trophic effects. PMID- 16382884 TI - Proliferative senescence in hematopoietic stem cells during ex-vivo expansion. AB - The good outcome of hematopoietic stem cell (HSC) transplantation is hampered by low doses of CD34+ cell infusion. Transplanted HSCs undergo a replicative stress that causes accelerated senescence due to rapid telomere shortening. The expansion of human cord blood HSCs is instrumental in obtaining a large number of "good quality" cells, in terms of telomere length and telomerase activity compared to adult HSCs. PMID- 16382885 TI - Basic fibroblast growth factor and its receptors in human embryonic stem cells. AB - Human embryonic stem cells (hESCs) are pluripotent stem cells with long-lasting capacity to self-renew and differentiate into various cell types of endodermal, ectodermal or mesodermal origin. Unlike mouse ESCs (mESCs), which can be maintained in an undifferentiated state simply by adding leukemia inhibitory factor (LIF) into the culture medium, hESCs are notorious for the sustained willingness to differentiate and not yet clearly defined signaling pathways that are crucial for their "stemness". Presently, our knowledge involves only limited number of growth factor signaling pathways that appear to be biologically relevant for stem cell functions in vitro. These include BMP, TGFbeta, Wnt, and FGF signaling pathway. The purpose of this review is to summarize recent data on the expression of FGFs and their receptors in hESCs, and critically evaluate the potential effects of FGF signals for their undifferentiated growth and/or differentiation in context with our current understanding of FGF/FGFR biology. PMID- 16382886 TI - Nonhematopoietic stem cells of fetal origin--how much of today's enthusiasm will pass the time test? AB - Stem cells originating at fetal age are for many reasons superior as a material for the regenerative medicine purposes, when compared to their adult counterparts. While hematopoietic cells, isolated from fetal liver or cord blood, have been well known for a long time and have passed practical tests as clinical transplantation material, the non-hematopoietic cells are newly recognized, and the knowledge of their phenotype and differentiation potential is rather insufficient. We, and the others, have identified a subpopulation of cord blood cells phenotypically different from hematopoietic cells (CD34-, CD45-, CD29+, CD44+, CD51+, CD105+, SH-2, SH-3), in vitro plastic adherent, and capable of multilineage differentiation. The other candidates for multipotential stem cells are cells extracted from umbilical cord or placental tissue. The preliminary observations suggest, that these cells, phenotypically similar to the nonhematopoietic cord blood cells, are capable of extensive replication in vitro and of multilineage differentiation into a variety of tissues including cardiac muscle, bone and cartilage, adipocytes, and nerve cells. The other possible medical applications include "rejuvenation" of selected tissues and systems in senile patients, and therapeutical cloning - for both purposes, cells at the fetal stage of genetic regulation may be more useful than cells collected from adult donors. There is still, however, a high level of uncertainty concerning future medical applications of fetal stem cells. Their numbers and characteristics may differ from the preliminary observations, and their behavior in vivo may not fulfill the expectations originating from the in vitro studies. Finally, the autologous applications of stem cells collected at the stage of birth may need the involvement of technical and financial resources for the storage of frozen cell samples throughout the period of life of their potential user. Such procedure seems possible from technical point of view, but may be inadequately substantiated by the eventual advantages. PMID- 16382887 TI - Messenger RNA electroporation: an efficient tool in immunotherapy and stem cell research. AB - Over the last decades medicine has developed tremendously, but still many diseases are incurable. The last years, cellular (gene) therapy has become a hot topic in biomedical research for the potential treatment of cancer, AIDS and diseases involving cell loss or degeneration. Here, we will focus on two major areas within cellular therapy, cellular immunotherapy and stem cell therapy, that could benefit from the introduction of neo-expressed genes through mRNA electroporation for basic research as well as for clinical applications. For cellular immunotherapy, we will provide a state-of-the-art on loading antigen presenting cells with antigens in the mRNA format for manipulation of T cell immunity. In the area of stem cell research, we will highlight current gene transfer methods into adult and embryonic stem cells and discuss the use of mRNA electroporation for controlling guided differentiation of stem cells into specialized cell lineages. PMID- 16382888 TI - Differentiation potential of the fetal rat liver-derived cells. AB - Mesenchymal stem cells derived from bone marrow or several fetal tissues can be expanded and differentiated into other cell lines. The fetal liver is the source of early hematopoietic cells and also, as a fetal tissue, may be considered as a source of pluripotent stem cells. The differentiation potential of fetal rat liver cells have been examined. Freshly isolated liver cells from 14-d fetuses were cultured in Dulbecco medium supplemented with 10% FCS. The plastic-adherent cells were then passaged up to 10 times. Freshly isolated cells and cells from every passage were cultured in hematopoiesis-promoting environment that consists of methylcelulose supplemented with FCS, rat IL-3, human IL-6 and Epo. Parallely these cells were incubated in co-culture with rat muscle satellite cells (Dulbecco medium with 10% FCS and 10% HS) to examine their myogenic potential. Culture in methylcelulose resulted in a high number of GM and Mix colonies in case of freshly isolated liver cells and the number of colonies decreased according to the number of passages. In case of cells from 4th passage, there ware no hematopoietic colonies in culture. In contrast--freshly isolated cells were not able to fuse with rat satellite cells and form the myotubes. This ability appeared in plastic-adherent cells just from the second passage and increases to 5th passage. The cells from every next passage up to 10th when co cultured with satellite cells participated in myotube formation at the same high level. This result may suggest that in the 14-d rat liver there exist at least two subpopulations of cells: the non-adherent hematopoietic cell population, and the population of plastic-adherent cells capable of differentiating into myotubes. Since the attempts to redifferentiate hematopoietic subpopulation into myopoiesis, or myopoietic subpopulation into hematopoiesis failed, it may be concluded that at least under our experimental conditions the fetal liver cells do not reveal the "plasticity" features. PMID- 16382889 TI - Which place for stem cell therapy in the treatment of acute radiation syndrome? AB - Radiation-induced (RI) tissue injuries can be caused by radiation therapy, nuclear accidents or radiological terrorism. Notwithstanding the complexity of RI pathophysiology, there are some effective approaches to treatment of both acute and chronic radiation damages. Cytokine therapy is the main strategy capable of preventing or reducing the acute radiation syndrome (ARS), and hematopoietic growth factors (GF) are particularly effective in mitigating bone marrow (BM) aplasia and stimulating hematopoietic recovery. However, first, as a consequence of RI stem and progenitor cell death, use of cytokines should be restricted to a range of intermediate radiation doses (3 to 7 Gy total body irradiation). Second, ARS is a global illness that requires treatment of damages to other tissues (epithelial, endothelial, glial, etc.), which could be achieved using pleiotropic or tissue-specific cytokines. Stem cell therapy (SCT) is a promising approach developed in the laboratory that could expand the ability to treat severe radiation injuries. Allogeneic hematopoietic stem cell transplantation (BM, mobilized peripheral blood and cord blood) transplantation has been used in radiation casualties with variable success due to limiting toxicity related to the degree of graft histocompatibility and combined injuries. Ex vivo expansion should be used to augment cord blood graft size and/or promote very immature stem cells. Autologous SCT might also be applied to radiation casualties from residual hematopoietic stem and progenitor cells (HSPC). Stem cell plasticity of different tissues such as liver or skeletal muscle, may also be used as a source of hematopoietic stem cells. Finally, other types of stem cells such as mesenchymal, endothelial stem cells or other tissue committed stem cells (TCSC), could be used for treating damages to nonhematopoietic organs. PMID- 16382890 TI - Mobilization of bone marrow-derived progenitor cells in acute coronary syndromes. AB - Two hypotheses explain the role of adult progenitor cells in myocardial regeneration. Stem cell plasticity which involves mobilization of stem cells from the bone marrow and other niches, homing to the area of tissue injury and transdifferentiation into functional cardiomyocytes. Alternative hypothesis is based on the observations that bone marrow harbors a heterogenous population of cells positive for CXCR4 - receptor for chemokine SDF-1. This population of non hematopoietic cells expresses genes specific for early muscle, myocardial and endothelial progenitor cells (EPC). These tissue-committed stem cells circulate in the peripheral blood at low numbers and can be mobilized by hematopoietic cytokines in the setting of myocardial ischemia. Endothelial precursors capable of transforming into mature, functional endothelial cells are present in the pool of peripheral mononuclear cells in circulation. Their number significantly increases in acute myocardial infarction (AMI) with subsequent decrease after 1 month, as well as in patients with unstable angina in comparison to stable coronary heart disease (CHD). There are numerous physiological and pathological stimuli which influence the number of circulating EPC such as regular physical activity, medications (statins, PPAR-gamma agonists, estrogens), as well as numerous inflammatory and hematopoietic cytokines. Mobilization of stem cells in AMI involves not only the endothelial progenitors but also hematopoietic, non hematopoietic stem cells and most probably the mesenchymal cells. In healthy subjects and patients with stable CHD, small number of circulating CD34+, CXCR4+, CD117+, c-met+ and CD34/CD117+ stem cells can be detected. In patients with AMI, a significant increase in CD34+/CXCR4+, CD117+, c-met+ and CD34/CD117+ stem cell number the in peripheral blood was demonstrated with parallel increase in mRNA expression for early cardiac, muscle and endothelial markers in peripheral blood mononuclear cells. The maximum number of stem cells was found early in ST-segment elevation myocardial infarction (<12 hours) with subsequent decrease through the 7-day follow-up and with concomitant changes in the levels of cytokines involved in the inflammatory response and stem cell recruitment. Moreover, peak expression of cardiac muscle and endothelial markers occurred at the same time as the most significant increase in CD34/CXCR4+ stem cell number. The SDF-1/CXCR-4 axis seems particularly important in stem/muscle progenitor cell homing, chemotaxis, engraftment and retention in ischaemic myocardium. The significance of autologous stem cells mobilization in terms of cardiac salvage and regeneration needs to be proved in humans but it seems to be a reparative mechanism triggered early in the course of acute coronary syndromes. PMID- 16382891 TI - Clinical trials using autologous bone marrow and peripheral blood-derived progenitor cells in patients with acute myocardial infarction. AB - This paper discusses the current data concerning the results of major clinical trials using bone marrow-derived and peripheral blood-derived stem/progenitor cells in treatment of patients with acute myocardial infarction (AMI) and depressed left ventricular ejection fraction. In all major trials (TOPCARE-AMI, BOOST), the primary outcome measure was increase in left ventricular systolic function (LVEF) and left ventricle remodeling. The most consistent finding is the significant increase in LVEF. Some trials suggest also reduction of left ventricular remodeling. Although the absolute LVEF increase is small (6-9%), it may substantially contribute to the improvement of global LV contractility. None of the studies in AMI patients treated with intracoronary infusion of progenitor cells revealed excess risk of arrythmia, restenosis or other adverse effects attributable to the therapy. The exact mechanism of improved myocardial contractile function remains unknown, however, there are several possible explanations: therapeutic angiogenesis improving the blood supply to the infarct border zone, paracrine modulation of myocardial fibrosis and remodeling (e.g. inhibition of myocyte apoptosis) and transdifferentiation of stem/progenitor cells into functional cardiomyocytes. No study showed the superiority of the particular subpopulation of autologous progenitor cells in terms of left ventricular function improvement in AMI. In fact, most of the clinical trials used the whole population of mononuclear bone marrow-derived progenitor cells, peripheral blood derived progenitor cells (endothelial progenitors). PMID- 16382892 TI - Survivin antiapoptotic gene expression as a prognostic factor in non-small cell lung cancer: in situ hybridization study. AB - Survivin is an inhibitor of apoptosis that plays a significant role in cell cycle regulation and is important for survival prognosis in many neoplasms. Survivin expression was assessed by in situ hybridization (ISH) in 60 consecutive patients (54 males and 4 females) with NSCLC treated between 1993 and 1997. The examined patients had IIB and IIIA stage according to TNM system. In all cases the chemotherapy with cisplatin and etoposide (2 cycles) was administered prior the surgery; in patients responding to the therapy one more cycle was applied. Survivin gene overexpression was observed in 35 patients (58.3%). There was no correlation between survivin mRNA level and histological type of tumor, stage of cell differentiation, stage of disease according to TNM classification, performance status according to WHO and number of chemotherapy regimens administered (p > 0.05). However, the correlation between survivin gene expression and response to the chemotherapy was statistically significant (p = 0.04). Statistical analysis showed that median survival in patients with survivin gene overexpression was shorter (14.0 months) as compared to patients with no expression (60.0 months; p = 0.00002). In survival assessment by means of Kaplan Meier test, 14.3% of five-year survival was achieved in the former group versus 60% in the latter (p = 0.00003). Univariate analysis (log-rank test) showed that significant independent prognostic factors in NSCLC included: stage of the disease according to TNM classification (p = 0.006), response to chemotherapy (p = 0.005) and pattern of survivin gene expression (p = 0.00003). Multivariate analysis utilizing Cox's model showed that for survival assessment the stage according to TNM, response to the chemotherapy and survivin expression estimated by means of ISH are of statistical significance (p=0.00001). The calculated predictive values showed that ISH technique was quite accurate in assessment of five-year survival. Our data show that survivin expression may be used as a prognostic factor and a target for therapy. PMID- 16382893 TI - Ultrastructural features of supraspinal muscles in rabbits after long-term transcutaneous lateral electrical surface stimulation (LESS). AB - Lateral electrical surface stimulation is one of methods used in the therapy of the progressive form of idiopathic scoliosis (IS) in children and youth. However, there are data suggesting that this method may lead to serious adverse side effects, when used for a too long period of time per day. To clarify this issue, the present study was aimed at disclosing possible changes in the ultrastructural appearance of rabbit supraspinal muscles undergoing long-term stimulation (9 h per day, 3 months), an animal model successfully used to mimic the situation in humans. In comparison to the control animals, muscles of "overstimulated" rabbits exhibited clear signs of microscopical lesions, including depletion and disintegration of myofilaments, proliferation, dilatation and, sometimes, swelling of sarcoplasmic reticulum and/or mitochondria, as well as signs of destruction of the Z line. The above-mentioned abnormalities, especially the signs of degenerative processes associated with the Z line and the observed microlesions strongly suggest that the failure of the long-term LESS therapy of the IS may be attributable to these ultrastructural lesions. PMID- 16382894 TI - Localization of pectins and Ca2+ ions in unpollinated and pollinated wet (Petunia hybrida Hort.) and dry (Haemanthus albiflos L.) stigma. AB - The subcellular localization of Ca2+ ions as well as esterified and deesterified pectins in unpollinated and pollinated wet (Petunia hybrida) and dry (Haemanthus albiflos) stigma was analyzed. Stigmas with different surfaces were found to differ in Ca2+ and pectin localization. In a wet Petunia hybrida stigma, Ca2+ ions were present in the exudate occurring in the intercellular spaces of secretory tissue before pollination. The exudate of an unpollinated stigma was the site of the localization of large amounts of deesterified pectins. Stigma penetration by pollen tubes induced the lysis of this category of pectins. The epidermal cells walls of the dry Haemanthus albiflos stigma before pollination lacked free and loosely bound Ca2+ ions. Pollination induced an accumulation of these ions in the apoplast of the stigma epidermal cells. In cells walls of an unpollinated stigma, mainly esterified pectins were present. Their deesterification took place after pollination at the site of pollen grain adhesion and then at the site of pollen tube growth. These results have shown that wet and dry stigmas differ in pectin metabolism and in the mechanism of forming a calcium environment at the site of pollen grain germination. PMID- 16382895 TI - Solvent-free mass spectrometry for hydrophobic peptide sequence analysis and protein conformation studies. PMID- 16382896 TI - SNAP-tag mediated live cell labeling as an alternative to GFP in anaerobic organisms. PMID- 16382897 TI - Brighter reporter genes from multimerized fluorescent proteins. PMID- 16382898 TI - Tissue microarray construction from bone marrow biopsies. PMID- 16382899 TI - Adenoviral transgene expression enhanced by cotreatment with etoposide in cultured cells. PMID- 16382900 TI - Alginate encapsulation is a highly reproducible method for tumor cell implantation in dorsal skinfold chambers. PMID- 16382901 TI - Combining restriction digestion and touchdown PCR permits detection of trace isoforms of histamine H3 receptor. AB - The conserved sequences of the mouse histamine H3 receptor at the potential alternative splice junctions suggest that the splice isoforms found in guinea pig, rat, human, and hamster may also be present in the mouse. However, the trace amount isoforms are hard to be detected by the regular PCR approach. In this paper, we report a method in which the unspliced long isoform is cut by restriction endonuclease so that the short isoforms can be amplified to detectable levels to confirm the existence of the splice isoforms of H3 receptor mRNA in the mouse. This method is applicable to the detection of trace amounts of splice isoforms that coexist with the long, more abundant isoforms. PMID- 16382902 TI - Bulk and micropatterned conjugation of extracellular matrix proteins to characterized polyacrylamide substrates for cell mechanotransduction assays. AB - Increasing numbers of cell mechanotransduction studies are currently utilizing elastic substrates fabricated from polyacrylamide in the form of thin gels. Their versatility depends on the ability to ensure the appropriate gel stiffness and control the uniformity and geometry of extracellular matrix protein coating of the gel. Beginning with a brief quantitative emphasis on the elastic properties of polyacrylamide gels, we present an inexpensive and highly reproducible method for uniform coating with a wide variety of extracellular matrix proteins. We used a reducing agent, hydrazine hydrate, to modify nonreactive amide groups in polyacrylamide to highly reactive hydrazide groups that can form covalent bonds with aldehyde or ketone groups in oxidized proteins. This simple conjugation method overcomes the limitations of previously used photoactivatable cross linkers: nonuniform coating due to nonuniformity of irradiation and technically challenging procedures for micropatterning. As demonstrated in our study of cell polarity during constrained migration, this conjugation method is especially effective in gel micropatterning by manual microcontact printing of protein patterns as small as 5 microm and enables numerous studies of constrained cell attachment and migration that were previously unfeasible due to high cost or difficulty in controlling the protein coating. PMID- 16382903 TI - Determination of allele frequency in pooled DNA: comparison of three PCR-based methods. AB - Determination of allele frequency in pooled DNA samples is a powerful and efficient tool for large-scale association studies. In this study, we tested and compared three PCR-based methods for accuracy, reproducibility, cost, and convenience. The methods compared were: (i) real-time PCR with allele-specific primers, (ii) real-time PCR with allele-specific TaqMan probes, and (iii) quantitative sequencing. Allele frequencies of three single nucleotide polymorphisms in three different genes were estimated from pooled DNA. The pools were made of genomic DNA samples from 96 cases with basal cell carcinoma of the skin and 96 healthy controls with known genotypes. In this study, the allele frequency estimation made by real-time PCR with allele-specific primers had the smallest median deviation (MD) from the real allele frequency with 1.12% (absolute percentage points) and was also the cheapest method. However; this method required the most time for optimization and showed the highest variation between replicates (SD = 6.47%). Quantitative sequencing, the simplest method, was found to have intermediate accuracies (MD = 1.44%, SD = 4.2%). Real-time PCR with TaqMan probes, a convenient but very expensive method, had an MD of 1.47% and the lowest variation between replicates (SD = 3.18%). PMID- 16382904 TI - Software for quantification of labeled bacteria from digital microscope images by automated image analysis. AB - Automated image analysis software, CellC, was developed and validated for quantification of bacterial cells from digital microscope images. CellC enables automated enumeration of bacterial cells, comparison of total count and specific count images [e.g., 4',6-diamino-2-phenylindole (DAPI) and fluorescence in situ hybridization (FISH) images], and provides quantitative estimates of cell morphology. The software includes an intuitive graphical user interface that enables easy usage as well as sequential analysis of multiple images without user intervention. Validation of enumeration reveals correlation to be better than 0.98 when total bacterial counts by CellC are compared with manual enumeration, with all validated image types. The software is freely available and modifiable: the executable files and MATLAB source codes can be obtained at www. cs. tut.fi/sgn/csb/cellc. PMID- 16382905 TI - Laser capture microdissection of bacterial cells targeted by fluorescence in situ hybridization. AB - Direct cultivation-independent sequence retrieval of unidentified bacteria from histological tissue sections has been limited by the difficulty of selectively isolating specific bacteria from a complex environment. Here, a new DNA isolation approach is presented for prokaryotic cells. By this method, a potentially pathogenic strain of the genus Brachyspira from formalin-fixed human colonic biopsies were visualized by fluorescence in situ hybridization (FISH) with a 16S rRNA-targeting oligonucleotide probe, followed by laser capture microdissection (LCM) of the targeted cells. Direct 16S rRNA gene PCR was performed from the dissected microcolonies, and the subsequent DNA sequence analysis identified the dissected bacterial cells as belonging to the Brachyspira aalborgi cluster 1. The advantage of this technique is the ability to combine the histological recognition of the specific bacteria within the tissue with molecular analysis of 16S rRNA gene or other genes of interest. This method is widely applicable for the identification of noncultivable bacteria and their gene pool from formalin fixed paraffin-embedded tissue samples. PMID- 16382906 TI - Handling three regulatory elements in one transgene: combined use of cre-lox, FLP FRT, and I-Scel recombination systems. AB - Studies of regulatory systems in transgenic Drosophila are often compromised by possible genomic position effects on gene expression. As a result, it is desirable to be able to manipulate multiple regulatory elements in a single transgene construct. We developed an I-SceI endonuclease-based method to efficiently delete preassigned sequences from transgenes with the use of direct repeat sequences of just 126 nucleotides. This system can be used in combination with the existing cre-lox and FLP-FRT recombinational mechanisms in order to modify up to three regulatory regions in a given transgene. We validated the utility of our combination approach by demonstrating new properties of the Fab-7 insulator. PMID- 16382907 TI - Hydroxyapatite chromatography of phage-display virions. AB - Hydroxyapatite column chromatography can be used to purify filamentous bacteriophage--the phage most commonly used for phage display. Virions that have been partially purified from culture supernatant by two cycles of precipitation in 2% polyethylene glycol are adsorbed onto the matrix at a density of at least 7.6 x 10(13) virions (about 3 mg) per milliliter of packed bed volume in phosphate-buffered saline (PBS; 0.15 M NaCl, 5 mM NaH2PO4, pH-adjusted to 7.0 with NaOH). The matrix is washed successively with wash buffer I(150 mM NaCl, 125 mM phosphate, pH 7.0), wash buffer II (2.55 M NaCl, 125 mM phosphate, pH 7.0), and wash buffer I; after which virions are desorbed in desorption buffer (150 mM NaCl, 200 mM phosphate, pH 7.0), and the matrix is stripped with stripping buffer (150 mM NaCl, 1 Mphosphate, pH 7.0). About half of the applied virions are recovered in desorption buffer. Western blot analysis shows that they have undetectable levels of host-derived protein contaminants that are present in the input virions and in virions purified by CsCl equilibrium density gradient centrifugation--the method most commonly used to prepare virions in high purity. Hydroxyapatite chromatography is thus an attractive alternative method for purifying filamentous virions, particularly when the scale is too large for ultracentrifugation to be practical. PMID- 16382908 TI - High-throughput SNP genotyping by single-tube PCR with Tm-shift primers. AB - Despite many recent advances in high-throughput single nucleotide polymorphism (SNP) genotyping technologies, there is still a great need for inexpensive and flexible methods with a reasonable throughput. Here we report substantial modifications and improvements to an existing homogenous allele-specific PCR based SNP genotyping method, making it an attractive new option for researchers engaging in candidate gene studies or following up on genome-wide scans. In this advanced version of the melting temperature (Tm)-shift SNP genotyping method, we attach two GC-rich tails of different lengths to allele-specific PCR primers, such that SNP alleles in genomic DNA samples can be discriminated by the Tms of the PCR products. We have validated 306 SNP assays using this method and achieved a success rate in assay development of greater than 83% under uniform PCR conditions. We have developed a standalone software application to automatically assign genotypes directly from melting curve data. To demonstrate the accuracy of this method, we typed 592 individuals for 6 SNPs and showed a high call rate (>98%) and high accuracy (>99.9%). With this method, 6-10,000 samples can be genotyped per day using a single 384-well real-time thermal cycler with 2-4 standard 384-well PCR instruments. PMID- 16382909 TI - High-throughput phagocytosis assay utilizing a pH-sensitive fluorescent dye. AB - We describe a development of a novel high-throughput phagocytosis assay based on a pH-sensitive cyanine dye, CypHer5E, which is maximally fluorescent in an acidic environment. This dye is ideally suited for the study of phagocytosis because of the acidic conditions generated in the intracellular phagocytic vesicles after particle uptake. Use of CypHer5E-labeled particles results in greatly reduced background from noninternalized particles and makes the assay more robust. Additionally, CypHer5E-labeled particles are resistant to fluorescence quenching observed in the aggressive and acidic environment of the phagosome with traditional dyes. The CypHer5E-based assay has been shown to work reliably in a variety of cell types, including primary human monocytes, primary human dendritic cells, primary human endothelial cells, human monocytic THP-1 cell line, and human/mouse hybrid macrophage cell line WBC264-9C. Inhibition of CypHer5E bead uptake by cytochalasin D was studied, and the 50% inhibition concentration (IC50) was determined. The assay was performed in 96- and 384-well formats, and it is appropriate for high-throughput cellular screening of processes and compounds affecting phagocytosis. The CypHer5E phagocytosis assay is superior to existing protocols because it allows easy distinction of true phagocytosis from particle adherence and can be used in microscopy-based measurement of phagocytosis. PMID- 16382910 TI - Environmental health writ large. PMID- 16382911 TI - "Grassroots" global warming study. PMID- 16382912 TI - Nanoparticle agglomeration restricts uptake into living cells. PMID- 16382914 TI - Congress weakens environmental laws with loopholes in hurricane-recovery legislation. PMID- 16382913 TI - Distinguishing between sources of sewage contamination. PMID- 16382915 TI - Sunita Narain. Interview by Catherine M Cooney. PMID- 16382916 TI - Cold-water laundry detergent is a hot idea. PMID- 16382917 TI - EPA proposes to relax TRI reporting rules. PMID- 16382918 TI - Chemical reactivity as a tool for estimating persistence. PMID- 16382919 TI - Molecular structure in soil humic substances: the new view. AB - A critical examination of published data obtained primarily from recent nuclear magnetic resonance spectroscopy, X-ray absorption near-edge structure spectroscopy, electrospray ionization-mass spectrometry, and pyrolysis studies reveals an evolving new view of the molecular structure of soil humic substances. According to the new view, humic substances are collections of diverse, relatively low molecular mass components forming dynamic associations stabilized by hydrophobic interactions and hydrogen bonds. These associations are capable of organizing into micellar structures in suitable aqueous environments. Humic components display contrasting molecular motional behavior and may be spatially segregated on a scale of nanometers. Within this new structural context, these components comprise any molecules intimately associated with a humic substance, such that they cannot be separated effectively by chemical or physical methods. Thus biomolecules strongly bound within humic fractions are by definition humic components, a conclusion that necessarily calls into question key biogeochemical pathways traditionally thought to be required for the formation of humic substances. Further research is needed to elucidate the intermolecular interactions that link humic components into supramolecular associations and to establish the pathways by which these associations emerge from the degradation of organic litter. PMID- 16382920 TI - Improved accounting of emissions from utility energy storage system operation. AB - Several proposed utility-scale energy storage systems in the U.S. will use the spare output capacity of existing electric power systems to create the equivalent of new load-following plants that can rapidly respond to fluctuations in electricity demand and increase the flexibility of baseload generators. New energy storage systems using additional generation from existing plants can directly compete with new traditional sources of load-following and peaking electricity, yet this application of energy storage is not required to meet many of the Clean Air Act standards required of new electricity generators (e.g., coal or gas-fired power plants). This study evaluates the total emissions that will likely result from the operation of a new energy storage facility when coupled with an average existing U.S. coal-fired power plant and estimates that the emission rates of SO2 and NOx will be considerably higher than the rate of a new plant meeting Clean Air Act standards, even accounting for the efficiency benefits of energy storage. This study suggests that improved emissions "accounting" might be necessary to provide accurate environmental comparisons between energy storage and more traditional sources of electricity generation. PMID- 16382921 TI - Learning from the U.S. National Assessment of Climate Change Impacts. AB - The U.S. National Assessment of the Potential Consequences of Climate Variability and Change was a federally coordinated nationwide effort that involved thousands of experts and stakeholders. To draw lessons from this effort, the 10 authors of this paper, half of whom were not involved in the Assessment, developed and administered an extensive survey, prepared a series of working papers, and conducted an invitational workshop in Washington, DC, on April 29, 2004. Considering all these sources, the authors conclude that the Assessment was largely successful in implementing its basic design of distributed stakeholder involvement and in achieving its basic objectives. Future assessments could be significantly improved if greater attention were devoted to developing a collective understanding of objectives, preparing guidance materials and providing training for assessment participants, developing a budgeting mechanism which would allow greater freedom in allocating resources across various assessment activities, and creating an environment in which assessments were part of an ongoing process. PMID- 16382922 TI - Effects of river flooding on PCDD/F and PCB levels in cows' milk, soil, and grass. AB - This paper presents the results of a study examining whether the flooding of pasture by rivers gives rise to higher PCDD/F and PCB concentrations in cows' milk. Over 180 milk, soil, and grass samples, taken from 38 farms across 3 different river systems (River Dee, Trent, and Doe Lea/Rother/Don) in the United Kingdom, were analyzed for PCDD/Fs and PCBs. The concentrations were compared between flood-prone farms, where the animals had access to pasture that is often flooded, and control farms where the land does not flood. The results indicated that concentrations of PCDD/Fs and PCBs in cows' milk were higher in samples taken from farms prone to flooding, but only from the river systems flowing through industrial and urban areas. Raised levels of PCDD/F and PCBs were also found in soil and grass from farms prone to flooding providing strong corroborative evidence that the higher concentrations in cows' milk from such areas is likely to be due to the ingestion of contaminated grass and soil. Overall, the results provide strong evidence that flooding of pastureland can indeed result in elevated concentrations of PCDD/Fs and PCBs in milk from the farms so affected. PMID- 16382923 TI - Microbial incorporation of 13C-labeled acetate at the field scale: detection of microbes responsible for reduction of U(VI). AB - A field-scale acetate amendment experiment was performed in a contaminated aquifer at Old Rifle, CO to stimulate in situ microbial reduction of U(VI) in groundwater. To evaluate the microorganisms responsible for microbial uranium reduction during the experiment, 13C-labeled acetate was introduced into well bores via bio-traps containing porous activated carbon beads (Bio-Sep). Incorporation of the 13C from labeled acetate into cellular DNA and phospholipid fatty acid (PLFA) biomarkers was analyzed in parallel with geochemical parameters. An enrichment of active sigma-proteobacteria was demonstrated in downgradient monitoring wells: Geobacter dominated in wells closer to the acetate injection gallery, while various sulfate reducers were prominent in different downgradient wells. These results were consistent with the geochemical evidence of Fe(III), U(VI), and SO(4)2- reduction. PLFA profiling of bio-traps suspended in the monitoring wells also showed the incorporation of 13C into bacterial cellular lipids. Community composition of downgradient monitoring wells based on quinone and PLFA profiling was in general agreement with the 13C-DNA result. The direct application of 13C label to biosystems, coupled with DNA and PLFA analysis, PMID- 16382924 TI - Gaseous and particulate emissions from prescribed burning in Georgia. AB - Prescribed burning is a significant source of fine particulate matter (PM2.5) in the southeastern United States. However, limited data exist on the emission characteristics from this source. Various organic and inorganic compounds both in the gas and particle phase were measured in the emissions of prescribed burnings conducted at two pine-dominated forest areas in Georgia. The measurements of volatile organic compounds (VOCs) and PM2.5 allowed the determination of emission factors for the flaming and smoldering stages of prescribed burnings. The VOC emission factors from smoldering were distinctly higher than those from flaming except for ethene, ethyne, and organic nitrate compounds. VOC emission factors show that emissions of certain aromatic compounds and terpenes such as alpha and beta-pinenes, which are important precursors for secondary organic aerosol (SOA), are much higher from active prescribed burnings than from fireplace wood and laboratory open burning studies. Levoglucosan is the major particulate organic compound (POC) emitted for all these studies, though its emission relative to total organic carbon (mg/g OC) differs significantly. Furthermore, cholesterol, an important fingerprint for meat cooking, was observed only in our in situ study indicating a significant release from the soil and soil organisms during open burning. Source apportionment of ambient primary fine particulate OC measured at two urban receptor locations 20-25 km downwind yields 74 +/- 11% during and immediately after the burns using our new in situ profile. In comparison with the previous source profile from laboratory simulations, however, this OC contribution is on average 27 +/- 5% lower. PMID- 16382925 TI - Chlorinated, brominated, and perfluorinated contaminants in livers of polar bears from Alaska. AB - The existence of two subpopulations of polar bears in Alaska, the Beaufort Sea and the Chukchi Sea populations, has been documented. In this study, differences in concentrations and profiles of organochlorine pesticides, polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and perfluorinated acids were examined in livers of polar bears from the two subpopulations in Alaska. Concentrations of most of the organohalogens analyzed were greater in the Beaufort Sea subpopulation than in the Chukchi Sea subpopulation, except for HCHs and perfluorononanoic acid (PFNA), which were high in samples from the Chukchi Sea subpopulation. Concentrations of chlordanes, PCBs, and perfluorooctanesulfonate (PFOS) were significantly different between the two subpopulations. Chlordane was the predominant contaminant in the Beaufort Sea population, and PFOS was the major contaminant in the Chukchi Sea population. Polar bears from the Beaufort Sea showed significantly higher proportions of more highly chlorinated PCBs than those from the Chukchi Sea. Concentrations of several perfluorinated acids were significantly correlated. Overall, the concentrations and profiles of organohalogens analyzed in the two subpopulations of polar bears suggest differences in the sources of exposures between the two regions of Alaska. PMID- 16382926 TI - Effect of sewage-sludge application on concentrations of higher-brominated diphenyl ethers in soils and earthworms. AB - Polybrominated diphenyl ethers (PBDEs), including octa-decaBDEs, were found in soil and earthworm samples collected in 2000 from three research stations (reference plots and sewage-sludge-amended plots) and two farms (reference and amended/flooded soils) in Sweden. Sewage-sludge amendment at the research stations increased concentrations of all BDE congeners 2- to 13-fold, with the highest increases for BDE-209. Concentrations 100-to 1000-fold higher were seen in contaminated soils at both farms. BDE-209 was the predominant congener in all soils. sigmaPBDE concentrations in worms ranged from 3.1 to 38 000 ng/g lipid weight and were correlated to soil concentrations, including the octa-decaBDEs. Biota-soil accumulation factors declined in the following order: TeBDE > PeBDE > HxBDE > OcBDE > NoBDE > DeBDE, and ranged from 0.3 to 2 for the octa-decaBDEs. Thus, higher-brominated PBDEs, including BDE-209, are bioavailable from soils and accumulate in earthworms, presenting an exposure pathway into the terrestrial food web. High levels found at one farm 20 years after the last use of PBDEs indicate high persistence of PBDEs in soils, including BDE-209. No evidence of photolytic debromination of BDE-209 in soils was seen. PMID- 16382927 TI - Influence of climate change, tidal mixing, and watershed urbanization on historical water quality in Newport Bay, a saltwater wetland and tidal embayment in southern California. AB - Historical coliform measurements (n = 67,269; 32 years) in Newport Bay, a regionally important saltwater wetland and tidal embayment in southern California, have been compiled and analyzed. Coliform concentrations in Newport Bay decrease along an inland-to-ocean gradient, consistent with the hypothesis that this tidal embayment attenuates fecal pollution from inland sources. Nearly 70% of the variability in the coliform record can be attributed to seasonal and interannual variability in local rainfall, implying that stormwater runoff from the surrounding watershed is a primary source of coliform in Newport Bay. The storm loading rate of coliform from the San Diego Creek watershed--the largest watershed draining into Newport Bay--appears to be unaffected by the dramatic shift away from agricultural land-use that occurred in the watershed over the study period. Further, the peak loading of coliform during storms is larger than can be reasonably attributed to sources of human sewage, suggesting that nonhuman fecal pollution and/or bacterial regrowth contribute to the coliform load. Summer time measurements of coliform exhibit interannual trends, but these trends are site specific, apparently due to within-Bay variability in land-use, inputs of dry-weather runoff, and tidal mixing rates. Overall, these results suggest that efforts to improve water quality in Newport Bay will likely have greater efficacy during dry weather summer periods. Water quality during winter storms, on the other hand, appears to be dominated by factors outside of local management control; namely, virtually unlimited nonhuman sources of coliform in the watershed and global climate patterns, such as the El Nino Southern Oscillation, that modulate rainfall and stormwater runoff in southern California. PMID- 16382928 TI - Identifying pollutant sources in tidally mixed systems: case study of fecal indicator bacteria from marinas in Newport Bay, southern California. AB - This study investigates the contribution of several marinas to fecal indicator bacteria impairment in Newport Bay, a regionally important tidal embayment in southern California. Three different fecal indicator bacteria groups were assayed, including total coliform, Escherichia coli, and enterococci bacteria, all measured using the IDEXX Colilert and Enterolert system. To document temporal variability in the fecal indicator bacteria signal, water column samples (n = 4132) were collected from two marinas over time scales ranging from hours to months. To document spatial variability of the fecal indicator bacteria signal, water column and sediment samples were collected from a number of sites (n = 11 to 36, depending on the study) in and around the two marinas, over spatial scales ranging from meters to kilometers. To identify the dominant temporal and spatial patterns in these data a statistical approach--Empirical Orthogonal Function analysis--was utilized. Finally, to clarify the transport pathways responsible for the observed temporal and spatial patterns, fecal indicator bacteria data were compared to simultaneous measurements of tidal flow, temperature, and salinity. The results of this field effort collectively implicate runoff--both dry weather runoff at sampling sites located near some storm drains and wet weather runoff at all sites--as a primary source of fecal indicator bacteria in the water column and subtidal sediments. The results and analysis presented here reinforce the growing body of evidence that management of fecal indicator bacteria impairment in the coastal waters of southern California will require developing long-term strategies for treating nonpoint sources of both dry weather and stormwater runoff. PMID- 16382929 TI - Chemical heterogeneity of organic soil colloids investigated by scanning transmission X-ray microscopy and C-1s NEXAFS microspectroscopy. AB - Colloid release and deposition in soils and sorption of inorganic and organic pollutants to soil colloids are strongly influenced by the composition and chemical heterogeneity of colloidal soil particles. To investigate the chemical heterogeneity of organic soil colloids at the particle scale, we used synchrotron scanning transmission X-ray microscopy (STXM) and C-1s near-edge X-ray absorption fine structure (NEXAFS) spectroscopy on 49 individual particles isolated from the surface horizons of three forest soils. Stacks of 130 images of each particle were collected at different X-ray energies between 280 and 310 eV. From these image arrays, NEXAFS spectra were obtained for each pixel and analyzed by principle component analysis and cluster analysis (PCA-CA) to characterize the intraparticle heterogeneity of the organic components. The results demonstrate that the organic matter associated with water-dispersible soil colloids is chemically heterogeneous at the single-particle scale. PCA-CA identified at least two distinct regions within single particles. However, the spectral variations between these regions were much smaller than the variations of averaged NEXAFS spectra representing different particles from the same soil horizon, implying that interparticle heterogeneity is much larger than intraparticle heterogeneity. Especially the contents of aromatic and carboxyl carbon exhibited a large variability. Overall, the NEXAFS spectra of water-dispersible soil colloids were similar to the NEXAFS spectrum of the humic acid fraction, but differed clearly from the fulvic acid and dissolved organic matter fractions extracted from the same soil horizon using conventional techniques. PMID- 16382930 TI - Perfluorinated compounds in the plasma of loggerhead and Kemp's ridley sea turtles from the southeastern coast of the United States. AB - Perfluorinated compounds (PFCs) have been measured in blood of humans and wildlife and are considered globally distributed contaminants. We examined 12 PFCs in the plasma of 73 loggerhead sea turtles (Caretta caretta) and 6 Kemp's ridley sea turtles (Lepidochelys kempii) captured from inshore waters of Core Sound, North Carolina (NC), and offshore waters of South Carolina, Georgia, and Florida (SC-FL). Perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA) were the dominant compounds, with respective mean concentrations of 11.0 ng/mL and 3.20 ng/mL for loggerhead turtles and 39.4 ng/mL and 3.57 ng/mL for Kemp's ridley turtles. Mean PFOS concentrations were 2- to 12-fold higher than typical mean sigmaPCB concentrations (approximately 5 ng/g wet mass) measured previously in sea turtle blood. More than 79% of the samples had detectable levels of perfluorocarboxylates (PFCAs) with 8-12 carbons, whereas only 17% or less of samples had detectable levels of PFCAs with 6 or 7 carbons. No samples had detectable levels of PFCAs with 4 or 5 carbons. In loggerhead turtles, sigmaPFC concentrations were not influenced by sex (p > 0.05), but were higher in turtles captured from inshore waters of NC than in turtles from offshore waters of SC-FL (p = 0.009). A backward stepwise multiple regression model showed that sigmaPFC concentrations were (1) significantly higher in Kemp's ridley turtles than loggerhead turtles (p < 0.0001), (2) higher in larger turtles (p = 0.018; carapace length used as a proxy for age), and (3) higher in turtles captured toward the north (p = 0.006). These findings suggest that bioaccumulation of PFCs in sea turtles is influenced by species, age, and habitat. PMID- 16382931 TI - Source diagnostics of polycyclic aromatic hydrocarbons based on species ratios: a multimedia approach. AB - Often, the sources of polycyclic aromatic hydrocarbons (PAHs) in environmental media can be identified by comparing the ratios of concentrations of selected pairs of PAH congeners in the source emissions to the ratios in the contaminated environmental media. However, these ratios can be altered significantly due to differences in the transport of the PAH compounds in a multimedia environment. To examine such changes, a fugacity model was applied to PAH ratios in a model environment. A linear relationship between the rate of emission and the bulk media concentration was identified for each PAH compound in an environmental medium at steady state and was quantified by a receptor-to-source ratio (RRS). It was demonstrated that the RRS values of the two congeners usually differ significantly. Consequentially, PAH ratios changed remarkably from the source emissions to various environmental media. A site-specific rectification factor (RF) was defined as the ratio of the two RRS values of the paired congeners for a specific PAH ratio in a given medium, which can be applied to account for the ratio changes in a multimedia environment. The PAH ratio changes were further verified with the surface soil data collected from Tianjin, China, and the observed changes of PAH ratios were compared favorably with the model predictions. The sensitivity analysis revealed that PAH ratios of the low molecular weight compounds were less stable. The most influential parameters controlling PAH ratios were those pertaining to dry precipitation, surface-to-air diffusion, degradation in air and water, and exchange between water and sediment. PMID- 16382932 TI - Passive and active air samplers as complementary methods for investigating persistent organic pollutants in the Great Lakes Basin. AB - Data obtained using passive air samplers (PAS) are compared to active high-volume air sampling data in order to assess the feasibility of the PAS as a method, complementary to active high-volume air sampling (AAS), for monitoring levels of organochlorine (OC) pesticides, polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) in the Laurentian Great Lakes. PAS were deployed at 15 of the Integrated Atmospheric Deposition Network (IADN) sites on a quarterly basis between July 2002 and June 2003, and PAS and AAS results are compared. Levels for the OC pesticides are typically highest in agricultural areas, with endosulfan I dominating air concentrations with values ranging between 40 and 1090 pg x m(-3), dieldrin values between 15 and 165 pg x m(-3), and gamma-HCH values between 13 and 100 pg x m(-3). alpha-HCH was seen to be relatively uniform across the Great Lakes Basin with values ranging between 15 and 73 pg x m(-3). Large urban centers, such as Chicago and Toronto, have the highest levels of PCBs and PBDEs that range between 400 and 1200 pg x m(-3) and 10 and 70 pg x m(-3), respectively. Comparison of the AAS and the PAS data collected during this study shows good agreement, within a factor of 2 or 3, suggesting that the two sample methods produce comparable results. It is suggested that PAS networks, while providing data that are different in nature from AAS, can provide a cost-effective and complementary approach for monitoring the spatial and temporal trends of persistent organic pollutants. PMID- 16382933 TI - Spectroscopic study of carbaryl sorption on smectite from aqueous suspension. AB - Sorption of carbaryl (1-naphthyl-N-methyl-carbamate) from aqueous suspension to smectite was studied using Fourier transform infrared (FTIR), high-performance liquid chromatography (HPLC) (for batch sorption), and quantum chemical methods. The amount of carbaryl sorbed was strongly dependent on the surface-charge density of the smectite with more sorption occurring on the two "low" surface charge density smectites (SHCa-1 and SWy-2) compared to that of the high surface charge SAz-1 smectite. In addition, the amount of carbaryl sorbed was strongly dependent on the nature of the exchangeable cation and followed the order of Ba approximately Cs approximately Ca > Mg approximately K > Na approximately Li for SWy-2. A similartrend was found for hectorite (SHCa-1) of Cs > Ba > Ca > K approximately Mg > Na approximately Li. Using the shift of the carbonyl stretching band as an indicator of the strength of interaction between carbaryl and the exchangeable cation, the observed order was Mg > Ca > Ba approximately K > Na > Cs. The position of the carbonyl stretching band shifted to lower wavenumbers with increasing ionic potential of the exchangeable cation. Density functional theory predicted a cation-induced lengthening of the C=O bond, resulting from the carbonyl group interacting directly with the exchangeable cation in support of the spectroscopic observations. Further evidence was provided by a concomitant shift in the opposite direction by several vibrational bands in the 1355-1375 cm(-1) region assigned to stretching bands of the carbamate N-Ccarbonyl and Oether-Ccarbonyl bonds. These data indicate that carbaryl sorption is due, in part, to site-specific interactions between the carbamate functional group and exchangeable cations, as evidenced by the FTIR data. However, these data suggest that hydrophobic interactions also contribute to the overall amount of carbaryl sorbed. For example, the FTIR data indicated thatthe weakest interaction occurred when Cs+ was the exchangeable cation. In contrast, the highest amount of carbaryl sorption was observed on Cs-exchanged smectite. Of all the cations studied, Cs has the lowest enthalpy of hydration. It is suggested that this low hydration energy provides the carbaryl with greater access to the hydrophobic regions of the siloxane surface. PMID- 16382934 TI - Quantification of particle number emission factors for motor vehicles from on road measurements. AB - The database on particle number emission factors has been very limited to date despite the increasing interest in the effects of human exposure to particles in the submicrometer range. There are also major questions on the comparability of emission factors derived through dynamometer versus on-road studies. Thus, the aims of this study were (1) to quantify vehicle number emission factors in the submicrometer (and also supermicrometer) range for stop-start and free-flowing traffic at about 100 km h(-1) driving conditions through extensive road measurements and (2) to compare the emission factors from the road measurements with those obtained previously from dynamometer studies conducted in Brisbane. For submicrometer particles the average emission factors for Tora Street were estimated at (1.89 +/- 3.40) x 10(13) particles km(-1) (mean +/- standard error; n = 386) for petrol and (7.17 +/- 2.80) x 10(14) particles km(-1) (diesel; n = 196) and for supermicrometer particles at 2.59 x 10(9) particles km(-1) and 1.53 x 10(12) particles km(-1), respectively. The average number emission factors for submicrometer particles estimated for Ipswich Road (stop-start traffic mode) were (2.18 +/- 0.57) x 10(13) particles km(-1) (petrol) and (2.04 +/- 0.24) x 10(14) particles km(-1) (diesel). One implication of the conclusion that emission factors of heavy duty diesel vehicles are over 1 order of magnitude higher than emission factors of petrol-fueled passenger cars is that future control and management strategies should in particular target heavy duty vehicles, as even a moderate decrease in emissions of these vehicles would have a significant impact on lowering atmospheric concentrations of particles. The finding that particle number emissions per vehicle-km are significantly larger for higher speed vehicle operation has an important implication on urban traffic planning and optimization of vehicle speed to lower their impact on airborne pollution. Additionally, statistical analysis showed that neither the measuring method (dynamometer or on road), nor data origin (Brisbane or elsewhere in the world), is associated with a statistically significant difference between the average values of emission factors for diesel, petrol, and vehicle fleet mix. However, statistical analyses of the effect of fuel showed that the mean values of emission factors for petrol and diesel are different at a 5% significance level. PMID- 16382935 TI - Fate and effects of enrofloxacin in aquatic systems under different light conditions. AB - The fate and effects of fluoroquinolone antibacterials (FQ) in the environment is of significance because of apparent increased FQ resistance in environmental and clinical organisms. Here we simultaneously assessed the fate and effects of enrofloxacin (enro), an FQ often used in agriculture, on the chemistry and in situ microbial communities in receiving waters. We added enro to 25 microg/L in nine outdoor mesocosms maintained under three light conditions (in triplicate): full sunlight typical of the upper epilimnion (100% full-light exposure, FLE), partial shading typical of the lower epilimnion (28% FLE), and near-complete shading typical of the hypolimnion (0.5% FLE). Enro disappearance and ciprofloxacin (cipro) formation were monitored over time using LC/MS, and water chemistry and ambient microbial communities (using denaturing gradient gel electrophoresis; DGGE) were characterized. Enro half-lives were 0.8, 3.7, and 72 days for the 100%, 28%, and 0.5% FLE treatments, respectively, creating three distinct FQ exposure scenarios. Although FQ exposures ranged from approximately 6 microg/L for 24 h to approximately 21 microg/L for 30 days, no statistically significant exposure effects were noted in water quality or microbial communities (as indicated by whole-community 16S rDNA DGGE analysis and specific amplification of the QRDR region of gyrase A). Small changes in water chemistry were noted over time; however, changes could not be specifically attributed to FQs. In general, enro addition had minimal effect on water column conditions at the levels and durations used here; however, further investigation is needed to assess effects in aquatic sediments. PMID- 16382936 TI - EXAFS analysis of arsenite adsorption onto two-line ferrihydrite, hematite, goethite, and lepidocrocite. AB - The modes of As(III) sorption onto two-line ferrihydrite (Fh), hematite (Hm), goethite (Gt), and lepidocrocite (Lp) have been investigated under anoxic condition using X-ray absorption spectroscopy (XAS). X-ray absorption near-edge structure spectroscopy (XANES) indicates that the absence of oxygen minimized As(III) oxidation due to Fenton reactions. Extended X-ray absorption fine structure spectroscopy (EXAFS) indicates thatAs(III)forms similar inner-sphere surface complexes on two-line ferrihydrite and hematite that differ from those formed on goethite and lepidocrocite. At high surface coverage, the dominant complex types on Fh and Hm are bidentate mononuclear edge-sharing (2E) and bidentate binuclear corner-sharing (2C), with As-Fe distances of 2.90 +/- 0.05 and 3.35 +/- 0.05 A, respectively. The same surface complexes are observed for ferrihydrite at low surface coverage. In contrast, As(III) forms dominantly bidentate binuclear corner-sharing (2C) sorption complexes on Gt and Lp [d(As-Fe) = 3.3-3.4 A], with a minor amount of monodentate mononuclear corner-sharing (1V) complexes [d(As-Fe) = 3.5-3.6 A]. Bidentate mononuclear edge-sharing (2E) complexes are virtually absent in Gt and Lp at the high surface coverages that were investigated in the present study. These results are compared with available literature data and discussed in terms of the reactivity of iron(III) (oxyhydr)oxide surface sites. PMID- 16382937 TI - Oxidation of gaseous elemental mercury to gaseous divalent mercury during 2003 polar sunrise at Ny-Alesund. AB - The springtime phenomenon, termed as the mercury depletion event (MDE), during which elemental gaseous mercury (Hg0) may be converted to a reactive form that accumulates in polar ecosystems, first noted in the Arctic, has now been observed at both poles and results in an important removal pathway for atmospheric mercury. An intensive international springtime mercury experiment was performed at Ny-Alesund, Spitsbergen, from 19 April to 13 May 2003 to study the atmospheric mercury chemistry in the Arctic environment and, in particular, the MDEs which occurred in the arctic boundary layer after polar sunrise. Automated ambient measurements of Hg0, divalent reactive gaseous mercury (RGM) and fine particulate mercury (<2.5 microm) (Hg(p)) were made at the Zeppelin Mountain Station (ZMS). During the experiment mercury concentrations in the lower atmosphere varied in synchrony with ozone levels throughout the Spring. Hg0 concentrations ranged from background levels (approximately 1.6 ng m(-3)) to undetectable values (<0.1 ng m( 3)) during the first and major MDE, while RGM data showed an opposite trend during the sampling period with concentrations increasing dramatically to a peak of 230 pg m(-3), synchronous with the depletion of Hg0. The results of a meteorological transport analysis indicate the MDEs observed at ZMS were primarily due to air masses being transported in from open water areas in the Arctic Ocean that were already depleted of Hg0 when they arrived and not due to in-situ oxidation mechanisms. PMID- 16382938 TI - Sorption of the antimicrobial ciprofloxacin to aluminum and iron hydrous oxides. AB - Solution chemistry (pH, ionic strength (I), and sorbate-to-sorbent ratio) effects on ciprofloxacin sorption to hydrous oxides of Al (HAO) and Fe (HFO) were investigated using macroscopic and spectroscopic analyses. Sorption to both HAO and HFO showed a strong pH-dependent behavior, following the fraction of zwitterionic species over the entire pH range studied. Increase in I from 0.01 to 0.5 M had an insignificant effect on the extent of ciprofloxacin sorption, and isotherms were well-described by the Langmuir model. HFO possessed a higher sorption capacity (0.066 mmol kg(-1)) than HAO (0.041 mmol kg(-1)). Ligand promoted dissolution of hydrous oxides, more pronounced for HAO, was observed in the presence of ciprofloxacin, but at a fairly high initial concentration (0.5 mM). Attenuated total reflectance Fourier transform infrared spectroscopy analysis indicated that different types of ciprofloxacin surface complexes are formed with HAO and HFO; while a monodentate mononuclear complex (with -COO-) appears likely between ciprofloxacin and HAO, keto O and one O from COO- seem to be involved in the formation of a six-membered ring with Fe on the HFO surface. The study results are expected to increase our understanding of the environmental reactivity of fluoroquinolones, an important class of antimicrobial compounds. PMID- 16382939 TI - Controls on arsenic speciation and solid-phase partitioning in the sediments of a two-basin lake. AB - Arsenic (As) regeneration from sediments of Spy Pond, a two-basin lake near Boston, MA, continues to result in seasonally elevated As levels despite the several decades that have elapsed since the pond's historical contamination by a pulse input of As. Solid-phase speciation and partitioning of As in the sediments appear to be primary determinants of both potential As regeneration rate and, conversely, the rate at which long-term burial of the pond's As burden will occur. Although iron (Fe) chemistry often controls As speciation in lakes, a higher rate of As regeneration in the south basin, accompanied by the absence of a correlation between regenerated As and Fe in this basin, suggest that additional factors operate to control As cycling in this lake. Solid-phase As speciation was remarkably similar between sediments of the two basins of this lake, about 40% of sedimentary As in upper sediments being in relatively labile (ionically bound and strongly adsorbed) fractions and a larger fraction of As being associated with recalcitrant minerals, likely sulfides. Extraction, X-ray fluorescence, and acid-volatile sulfide data collectively suggest that sedimentary As cycling in both basins is largely controlled by the formation of sulfide minerals. An accounting of the size of sulfur (S) pools supports this conclusion, showing that sufficient S exists in the north basin to control both Fe and As, while the south basin has lower S levels resulting in seasonal accumulation of Fe in the porewater. PMID- 16382940 TI - Photosensitizer method to determine rate constants for the reaction of carbonate radical with organic compounds. AB - Carbonate radical (CO3*-) is a powerful oxidant that is present in sunlit surface waters and in waters treated by advanced oxidation processes. The production of CO3*- in aqueous solution through oxidation of carbonate anion by excited triplet states of aromatic ketones was investigated in this study to provide new methods for the determination of rate constants and to explore a possible photoinduced pathway of CO3*- formation in the aquatic environment. Rate constants for triplet quenching by carbonate anion of up to 3.0 x 10(7) M(-1) s(-1) and CO3*- yields approaching unity, determined using laser flash photolysis, allowed us to conclude that such a formation mechanism might be significant in sulit natural waters. Kinetic methods based on either flash photolysis or steady-state irradiation and on the use of aromatic ketones as photosensitizers gave bimolecular rate constants in the range of 4 x 10(6) to 1 x 10(8) M(-1) s(-1) for the reaction of CO3*- with several s-triazine and phenylurea herbicides. For various anilines and phenoxide anions, rate constants determined by these methods agreed well with published values. Moreover, it could be shown for the first time by a direct method that dissolved natural organic matter (DOM) reduces the lifetime of CO3*- and a second-order rate constant of (280 +/- 90) (mg of C/L)( 1) s(-1) was obtained for Suwannee River fulvic acid. PMID- 16382941 TI - Effects of microbially mediated redox conditions on PAH-soil interactions. AB - The impacts of microbially mediated redox conditions on the bioavailability of persistent polycyclic aromatic hydrocarbons (PAHs) in soils and sediments have received little study, despite the fact that most water-saturated soils and sediments spend a significant portion of the time under reduced conditions. To address this need an uncontaminated surface soil was incubated under various redox conditions (aerobic, nitrate-reducing, sulfate-reducing, and methanogenic). Depending on redox conditions, different quantities of fulvic and humic acids were liberated as dissolved organic matter (DOM) from the soil during incubation. The DOM released under highly reduced conditions was more nonpolar, aromatic, and polydisperse, of higher molecular weight, and had a higher sorption capacity for pyrene compared to that obtained from relatively oxic incubations. The soil-phase organic matter incubated under reduced conditions also became relatively more aromatic, containing nonpolar organic molecules of lower oxygen contents and exhibiting higher capacity and more nonlinear and hysteric sorption/desorption behavior for pyrene. These observations support the hypothesis that reduced environments established by indigenous soil microbes alter soil organic matter in a matter similar to diagenetic processes. Such humification-like alteration occurred principally in relatively more labile fractions of soil organic matter. These findings are important for assessing the ultimate fate and exposure risk of hydrophobic organic contaminants in soils and sediments where living microorganisms play a significant role in formation and evolution of soil/sediment organic matter. PMID- 16382942 TI - Effect of precipitation on low frequency electrical properties of zerovalent iron columns. AB - We conducted column studies to investigate the application of a noninvasive electrical method to monitor precipitation in Fe0 columns using (a) Na2SO4 (0.01 M, dissolved oxygen (DO) = 8.8 ppm), and (b) Na2CO3 (0.01 M, DO = 2.3 ppm) solutions. An increase in complex conductivity terms (maximum 40% in sulfate column and 23% in carbonate column) occurred over 25 days. Scanning electron microscopy (SEM) identified mineral surface alteration, with greater changes in the high DO sulfate column relative to the low DO carbonate column. X-ray diffractometry (XRD) identified reduced amounts of hematite/maghemite in both columns, precipitation of goethite/akaganeite in the sulfate column, and precipitation of siderite in the carbonate column. Nitrogen adsorption measurements showed increases in specific surface area of iron minerals (27.5% for sulfate column and 8.2% for carbonate column). As variations in electrolytic conductivity and porosity were minimal, electrical changes are attributed to (1) higher complex interfacial conductivity due to increased surface area and mineralogical alteration and (2) increased electronic conduction due to enhanced electron transfer across the iron-fluid interface. Our results show that electrical measurements are a proxy indicator of Fe0 surface alteration. PMID- 16382943 TI - Phosphorus composition of sheep feces and changes in the field determined by 31P NMR spectroscopy and XRPD. AB - Information on the P species in sheep feces is lacking. Such information is required to understand P-cycling in grazed ecosystems. The P composition of feces from sheep grazing grass in Scotland was assessed on freeze-dried samples by 31P MAS (magic angle spinning) NMR (nuclear magnetic resonance) spectroscopy and XRPD (X-ray powder diffraction). The 31P MAS NMR spectrum showed resonances and sidebands consistent with dicalcium phosphate dihydrate (brushite) and ammonium magnesium phosphate hexahydrate (struvite). XRPD confirmed the result and allowed quantification of these minerals, which accounted for 63% of the P. To determine transformations in the field, sheep feces were collected and reapplied to sheep free pasture in synthetic patches during late summer. The dry weight decreased with time and the feces disappeared between 84 and 112 days following heavy rainfall. The concentration of P in the feces recovered at intervals up to 84 days changed little with time but the contribution from brushite and struvite decreased and within 1 week <50% remained indicating conversion into other forms. Solution-phase 31P NMR spectra of NaOH/EDTA extracts of the feces were dominated by the inorganic orthophosphate with minor amounts of organic P that were attributed to phosphate esters and polyphosphates. PMID- 16382944 TI - Acceleration and quenching of the photolysis of PCB in the presence of surfactant and humic materials. AB - The photodecay of a polychlorobiphenyl (PCB) congener, 2,3,4,5 tetrachlorobiphenyl (TeCB), in a surfactant-aided soil-washing process was investigated. The surfactant was found to be a useful hydrogen source in improving the TeCB photodecay via photoreduction process, while the light attenuation effect resulted from the nontarget compounds in the solution, and the impact of humic materials that co-extracted from the soil played important roles in the process of photolytic destruction of TeCB. A well-defined sediment, EPA 11, was used as the source of humic materials in examining the acceleration and quenching effects in the photolysis process. Experimental results indicated that the low dose of humic materials mainly acted as a supplementary hydrogen source to improve the photodegradation rate, while at a higher dose its amphoteric property of being a photochemical quencher become perceptible gradually, thus retarding the reaction. These effects were inspected and the dominant rate constants were quantified through the examination of possible sub-reactions in the assistant of proposed linear models in this study. The models under three different conditions at zero, low, and high humic levels associated with a light attenuation model were successfully developed to solve the dominant rate constants of the process. The good correlations between the experimental data and the models verify that the proposed reaction mechanisms of rate acceleration (resulting from the hydrogen source characteristics of the surfactant and humic material) and rate retardation (high levels of humic materials) coexist in the process. PMID- 16382945 TI - Iron(II)-catalyzed oxidation of arsenic(III) in a sediment column. AB - Arsenic contamination in aquatic systems is a worldwide concern. Understanding the redox cycling of arsenic in sediments is critical in evaluating the fate of arsenic in aquatic environments and in developing sediment quality guidelines. The direct oxidation of inorganic trivalent arsenic, As(III), by dissolved molecular oxygen has been studied and found to be quite slow. A chemical pathway for As(III) oxidation has been proposed recently in which a radical species, Fe(IV), produced during the oxidation of divalent iron, Fe(II), facilitates the oxidation of As(III). Rapid oxidation of As(III) was observed (on a time scale of hours) in batch systems at pH 7 and 7.5, but the extent of As(III) oxidation was limited. The Fe(II)-catalyzed oxidation of As(III) is examined in a sediment column using both computational and experimental studies. A reactive-transport model is constructed that incorporates the complex kinetics of radical species generation and Fe(II) and As(III) oxidation that have been developed previously. The model is applied to experimental column data. Results indicate that the proposed chemical pathway can explain As(III) oxidation in sediments and that transport in sediments plays a vital role in increasing the extent of As(III) oxidation and efficiency of the Fe(II) catalysis. PMID- 16382946 TI - Oxidative degradation of glyphosate and aminomethylphosphonate by manganese oxide. AB - Glyphosate (N-(phosphonomethyl)glycine), the most commonly used herbicide worldwide, degrades relatively rapidly in soils under most conditions, presumably by microbial processes. The most frequently detected degradation product in soil and water is AMPA (aminomethylphosphonic acid). We report the first evidence for an abiotic pathway of glyphosate and AMPA degradation under environmentally realistic conditions. Both glyphosate and AMPA degraded at 20 degrees C in dilute aqueous suspensions of birnessite, a manganese oxide common in soils, as evidenced by the accumulation of orthophosphate in solution over a period of several days. It is concluded that the abiotic degradation involved C-P bond cleavage at the Mn oxide surface, although evidence for C-N bond cleavage in the case of glyphosate and sarcosine, a likely degradation product of glyphosate, was found. The degradation of glyphosate was faster than that of AMPA, and higher temperature (50 degrees C) resulted in faster degradation of both glyphosate and AMPA. The addition of sulfate to the solution had no marked effect on the reaction rate, although Cu2+ addition inhibited degradation. As this metal ion complexes strongly with glyphosate, the inhibition can be attributed to the ability of Cu2+ to limit glyphosate coordination to reactive oxidation sites at the Mn oxide surface. Using a similar experimental design, we were unable to detect glyphosate degradation in an equimolar solution of MnCl2 (0.5 mM). However, we demonstrated that the oxidation of Mn2+ is enhanced both in solution and on an inert surface, in the presence of glyphosate (4:1 Mn-glyphosate molar ratio). This result suggests that the oxidative breakdown of glyphosate in the presence of Mn2+ may ultimately occur following the spontaneous oxygen-mediated oxidation of manganese. PMID- 16382947 TI - Products and mechanism of secondary organic aerosol formation from reactions of n alkanes with OH radicals in the presence of NOx. AB - Secondary organic aerosol (SOA) formation from reactions of n-alkanes with OH radicals in the presence of NOx was investigated in an environmental chamber using a thermal desorption particle beam mass spectrometer for particle analysis. SOA consisted of both first- and higher-generation products, all of which were nitrates. Major first-generation products were sigma-hydroxynitrates, while higher-generation products consisted of dinitrates, hydroxydinitrates, and substituted tetrahydrofurans containing nitrooxy, hydroxyl, and carbonyl groups. The substituted tetrahydrofurans are formed by a series of reactions in which sigma-hydroxycarbonyls isomerize to cyclic hemiacetals, which then dehydrate to form substituted dihydrofurans (unsaturated compounds) that quickly react with OH radicals to form lower volatility products. SOA yields ranged from approximately 0.5% for C8 to approximately 53% for C15, with a sharp increase from approximately 8% for C11 to approximately 50% for C13. This was probably due to an increase in the contribution of first-generation products, as well as other factors. For example, SOA formed from the C10 reaction contained no first generation products, while for the C15 reaction SOA was approximately 40% first generation and approximately 60% higher-generation products, respectively. First generation sigma-hydroxycarbonyls are especially important in SOA formation, since their subsequent reactions can rapidly form low volatility compounds. In the atmosphere, substituted dihydrofurans created from sigma-hydroxycarbonyls will primarily react with O3 or NO3 radicals, thereby opening reaction pathways not normally accessible to saturated compounds. PMID- 16382948 TI - Relationship between mercury accumulation in young-of-the-year yellow perch and water-level fluctuations. AB - A three-year (2001-2003) monitoring effort of 14 northeastern Minnesota lakes was conducted to document relationships between water-level fluctuations and mercury bioaccumulation in young-of-the-year (YOY) yellow perch (Perca flavescens) collected in the fall of each year at fixed locations. Six of those lakes are located within or adjacent to Voyageurs National Park and are influenced by dams on the outlets of Rainy and Namakan lakes. One site on Sand Point Lake coincides with a location that has nine years of previous monitoring suitable for addressing the same issue over a longer time frame. Mean mercury concentrations in YOY yellow perch at each sampling location varied significantly from year to year. For the 12-year monitoring site on Sand Point Lake, values ranged from 38 ng gww(-1) in 1998 to 200 ng gww(-1) in 2001. For the 14-lake study, annual mean concentrations ranged by nearly a factor of 2, on average, for each lake over the three years of record. One likely factor responsible for these wide variations is that annual water-level fluctuations are strongly correlated with mercury levels in YOY perch for both data sets. PMID- 16382949 TI - Use of the chiral pharmaceutical propranolol to identify sewage discharges into surface waters. AB - The discharge of relatively small volumes of untreated sewage is a source of wastewater-derived contaminants in surface waters that is often ignored because it is difficult to discriminate from wastewater effluent. To identify raw sewage discharges, we analyzed the two enantiomers of the popular chiral pharmaceutical, propranolol, after derivitization to convert the enantiomers to diastereomers. The enantiomeric fraction (the ratio of the concentration of one of its isomers to the total concentration) of propranolol in the influent of five wastewater treatment plants was 0.50 +/- 0.02, while after secondary treatment it was 0.42 or less. In a laboratory study designed to simulate an activated sludge municipal wastewater treatment system, the enantiomeric fraction of propranolol decreased from 0.5 to 0.43 as the compound underwent biotransformation. In a similar system designed to simulate an effluent-dominanted surface water, the enantiomeric fraction of propranolol remained constant as it underwent biotransformation. Analysis of samples from surface waters with known or suspected discharges of untreated sewage contained propranolol with an enantiomeric fraction of approximately 0.50 whereas surface waters with large discharges of wastewater effluent contained propranolol with enantiomeric fractions similar to those observed in wastewater effluent. Measurement of enantiomers of propranolol may be useful in detecting and documenting contaminants related to leaking sewers and combined sewer overflows. PMID- 16382950 TI - Automated speciated mercury measurements in Michigan. AB - Automated speciated mercury measurements were made at a rural (Dexter, MI) and an urban (Detroit, MI) site in Michigan during selected times from 1999 to 2002 to assess the concentrations of elemental (Hg0), reactive gaseous (RGM), and particulate mercury (Hgp) in these environments. Here we present the first-ever reported values for RGM in Michigan. Median RGM concentrations were 2.21-2.93 pg m(-3) at Dexter and were 3-11 times higher in Detroit at 6.41-22.0 pg m(-3). Maximum RGM concentrations of 38.7 pg m(-3) and 270 pg m(-3) were observed in Dexter and Detroit, respectively. Measured RGM/Hg0 ratios were in the range of 0.04-11.60% indicating that at times RGM comprises greater than the currently held view of 5% of total gaseous mercury in the air. Well-pronounced diurnal patterns of RGM were observed at the rural site, whereas the urban site exhibited patterns that were influenced by nighttime emissions and regional transport. An analysis of RGM/Hgp ratios at the urban site when combined with trajectory analysis suggests that the site receives mercury inputs from both local and regional sources. Episodes of elevated ozone concentrations which were accompanied by increases in RGM concentrations were observed to occur in the late afternoon and overnight. These may be evidence of advection of ozone and RGM over long distances to the site. PMID- 16382951 TI - Removal and inactivation of waterborne viruses using zerovalent iron. AB - A daunting challenge facing the water industry and regulators is how to simultaneously control microbial pathogens, residual disinfectant, and disinfection byproducts in drinking water, and to do so at an acceptable cost. Of the different pathogens, viruses are especially problematic due to their small size, high mobility, and resistance to chlorination and filtration. In the past decade, zerovalent iron has been used to treat a wide variety of organic and inorganic contaminants from groundwater. However, iron has not been tested against biological agents. This study examined the effectiveness of commercial zerovalent iron to remove two viruses, phiX174 and MS-2, from water. Removal of these viruses by iron granules in batch reactors was first-order, and the rate was likely controlled by external mass transfer. Most of the viruses removed from solution were either inactivated or irreversibly adsorbed to iron. In a flow through column containing zerovalent iron (with 20 min of iron contact time), the removal efficiency for both viruses was 4-log in an initial pulse test, and over 5-log in the second pulse test after passage of 320 pore volumes of artificial groundwater. We assume that the improved efficiency was due to continuous formation of new iron (oxyhydr)oxides which served as virus adsorption sites. To our knowledge, this is the first demonstration of biological agent removal from water by zerovalent iron. Results of this study suggest zerovalent iron may be potentially useful for disinfecting drinking water and wastewater, thereby reducing our dependence on chlorine and reducing the formation of disinfection byproducts. PMID- 16382952 TI - Sequential electrolytic oxidation and reduction of aqueous phase energetic compounds. AB - Contamination of soils and groundwater with energetic compounds has been documented at many former ammunition manufacturing plants and ranges. Recent research at Colorado State University (CSU) has demonstrated the potential utility of electrolytic degradation of organic compounds using an electrolytic permeable reactive barrier (e-barrier). In principle, an electrolytic approach to degrade aqueous energetic compounds such as hexahydro-1,3,5-trinitro-1,3,5 triazine (RDX) or 2,4,6-trinitrotoluene (TNT) can overcome limitations of management strategies that involve solely oxidation or reduction, through sequential oxidation-reduction or reduction-oxidation. The objective of this proof-of-concept research was to evaluate transformation of aqueous phase RDX and TNT in flow-through electrolytic reactors. Laboratory experiments were conducted using six identical column reactors containing porous media and expanded titanium mixed-metal-oxide electrodes. Three columns tested TNT transformation and three tested RDXtransformation. Electrode sequence was varied between columns and one column for each contaminant acted as a no-voltage control. Over 97% of TNT and 93% of RDX was transformed in the reactors under sequential oxidation-reduction. Significant accumulation of known degradation intermediates was not observed under sequential oxidation-reduction. Removal of approximately 90% of TNT and 40% of RDX was observed under sequential reduction-oxidation. Power requirements on the order of 3 W/m2 were measured during the experiment. This suggests that an in situ electrolytic approach may be cost-practical for managing groundwater contaminated with explosive compounds. PMID- 16382953 TI - Decomposition of 2,4,6-trinitrotoluene (TNT) by gamma irradiation. AB - The purpose of this study was to evaluate the potential of gamma irradiation to decompose 2,4,6-trinitrotoluene (TNT) in an aqueous solution; the concentration range of the TNT solution was 0.11-0.44 mmol/L. The decomposition rate of TNT by gamma irradiation was pseudo-first-order kinetic over the applied initial concentrations. The dose constant was strongly dependent on the initial concentration of TNT. Increasing the concentration of dissolved oxygen in the solution was more effective on the decomposition of TNT as well as its mineralization. The required irradiation dose to remove 90% of initial TNT (0.44 mmol/L) was 58, 41, 32, 28, and 25 kGy at the dissolved oxygen concentration of 0.025, 0.149, 0.3, 0.538, and 0.822 mmol/L, respectively. However, TOC still remained as 30% of the initial TOC (3.19 mmol/L) when 200 kGy irradiation dose was applied to the TNT solution (0.44 mmol/L) containing dissolved oxygen of 0.822 mmol/L. The removal of the TNT was more efficient at a pH below 3 and at a pH above 11 than at neutral pH (pH 5-9). The required irradiation dose to remove over 99% of the initial TNT (0.44 mmol/L) was 39, 76, and 10 kGy at pH 2, 7, and 13, respectively. The dose constant was increased 1.6-fold and over 15.6-fold at pH 2 and 13, respectively, compared to that at pH 7. When an irradiation dose of 200 kGy was applied, the removal efficiencies of the TOC (initial concentration 3.19 mmol/L) were 91, 46, and 53% at pH 2, 7, and 13, respectively. Ammonia and nitrate were detected as the main nitrogen byproducts of TNT, and glyoxalic acid and oxalic acid were detected as organic byproducts. PMID- 16382954 TI - Reductive biotransformation of tetrachloroethene to ethene during anaerobic degradation of toluene: experimental evidence and kinetics. AB - Reductive biotransformation of tetrachloroethene (PCE) to ethene occurred during anaerobic degradation of toluene in an enrichment culture. Ethene was detected as a dominant daughter product of PCE dechlorination with negligible accumulation of other partially chlorinated ethenes. PCE dechlorination was linked to toluene degradation, as evidenced by the findings that PCE dechlorination was limited in the absence of toluene but was restored with a spike of toluene again in the cultures. PCE was effectively dechlorinated in cultures amended with a wide range of concentrations of PCE and toluene. PCE dechlorination can be described by a Monod-like equation but followed a zero-order kinetic at high levels of PCE. In addition to toluene, benzoate and lactate were also able to be used as sole electron donors for reductive dechlorination of PCE in the cultures. In terms of dechlorination rates, lactate was the best electron donor followed by benzoate and then toluene. The kinetic characteristics of PCE dechlorination were retained in the cultures regardless of electron donors used, but the kinetic constant values were unique to each electron donor. The dechlorination rate was found to be closely correlated with the level of H2 produced during fermentation of the three organic compounds. Nitrate and sulfate were observed to be favorable electron acceptors in this culture, and their presence completely blocked electron flow to PCE. However, the presence of nitrate and sulfate did not destroy the capability of PCE dechlorination by the culture. PCE dechlorination was immediately reestablished after depletion of nitrate and sulfate in the culture. This anaerobic process provides an opportunity for concurrent remediation of chlorinated solvents and certain fuel hydrocarbons, and recognition of this process is also important in understanding the subsurface fate and transport of these contaminants under natural conditions. PMID- 16382955 TI - Chemical pathway and kinetics of phenol oxidation by Fenton's reagent. AB - Phenol oxidation by Fenton's reagent (H2O2 + Fe2+) in aqueous solution has been studied in depth for the purpose of learning more about the reactions involved and the extent of the oxidation process, under various operating conditions. An initial phenol concentration of 100 mg/L was used as representative of a phenolic industrial wastewater. Working temperatures of 25 and 50 degrees C were tested, and the initial pH was set at 3. The H2O2 and the Fe2+ doses were varied in the range of 500-5000 and 1-100 mg/L, respectively, corresponding to 1-10 times the stoichiometric ratio. A series of intermediates were identified, corresponding mainly to ring compounds and short-chain organic acids. Most significant among the former were catechol, hydroquinone, and p-benzoquinone; the main organic acids were maleic, acetic, oxalic, and formic, with substantially lower amounts of muconic, fumaric, and malonic acids. Under milder operating conditions (H2O2 and Fe2+ at lower concentrations), a great difference was found between the measured total organic carbon (TOC) and the amount of carbon in all analyzed species in the reaction medium. This difference decreased as the doses of H2O2 and Fe2+ increased, indicating that the unidentified compounds must correspond to oxidation intermediates between phenol and the organic acids. To establish a complete oxidation pathway, experiments were carried out using each of the identified intermediates as starting compounds. Dihydroxybenzenes were identified in the earlier oxidation stages. Muconic acid was detected in catechol but not in the hydroquinone and p-benzoquinone oxidation runs; the last two compounds were oxidized to maleic acid. Oxalic and acetic acid appeared to be fairly refractory to this oxidation treatment. A detailed knowledge of the time evolution of the oxidation intermediates is of environmental interest particularly in the case of hydroquinone and p-benzoquinone because their toxicities are several orders of magnitudes higher than that of phenol itself. The time evolution of the intermediates and TOC was fitted to a simple second-order kinetic equation, and the values of the kinetic constants were determined. This provides a simplified approach useful for design purposes. PMID- 16382956 TI - Reaction of nonaqueous phase TCE with permanganate. AB - Oxidative treatment of trichloroethylene (TCE) in the form of dense nonaqueous phase liquid (DNAPL) by potassium permanganate (KMnO4) was investigated in a series of batch tests. The study focused on understanding the fundamental mechanisms of oxidative removal of DNAPL TCE by permanganate oxidation. Dissolution experiment for DNAPL TCE has been performed as a control experiment in the absence of KMnO4. DNAPL TCE dissolved into the aqueous phase until it reached the saturation concentration of 1200 mg/L (9.16 x 10(-3) M) at 20 degrees C. The rate of dissolution of DNAPL TCE was proportional to the volume of the DNAPL. In the presence of KMnO4, the experimental results showed that the amount of TCE oxidized during the reaction was increased continuously as [MnO4-] decreased even though the rate decreased as [MnO4-] decreased. It was apparent that more DNAPL TCE was removed with a faster rate for higher initial permanganate concentration. At high permanganate concentration, the aqueous concentration of TCE was kept low and practically constant by the chemical reaction between aqueous TCE and MnO4-. However, as MnO4- was consumed in the system, the aqueous concentration started to increase until it reached solubility. From experimental observation, 1.56-1.78 mol of MnO4- was consumed per mole of TCE oxidized. Furthermore, 2.85-2.98 mol of Cl- was released to the solution per mole of TCE oxidized. Since the complete mineralization of TCE requires 2.0 mol of MnO4- and releases 3 mol of Cl- per mol of TCE oxidized, the observed stoichiometric factors indicated incomplete mineralization of TCE, but nearly complete dechlorination. Enhancement factor due to chemical reaction was quantified experimentally. The enhancement factor was shown to be a function of the molar ratio of MnO4- to TCE in the system, and hence varied during the reaction period. PMID- 16382957 TI - Mineralogy of air-pollution-control residues from a secondary lead smelter: environmental implications. AB - The mineralogy and solubility of air-pollution-control (APC) residues from a secondary lead (Pb) smelter have been studied on samples from the Pribram smelter, Czech Republic, recycling car batteries, with the emphasis on their potential environmental effect. The presence of dominant anglesite (PbSO4) and laurionite (Pb(OH)Cl) was observed in a sintered residue from after-burning chambers (800-1000 degrees C). In contrast, low-temperature Pb-bearing phases, such as KCl x 2PbCl2 and caracolite (Na3Pb2(SO4)3Cl), were detected in the major APC residue from bag-type fabric filters. Metallic elements, zinc (Zn), cadmium (Cd), and tin (Sn) were found homogeneously distributed within this residue. The formation of anglesite, cotunnite (PbCl2), (Zn,Cd)2SnO4, and (Sb,As)2O3 was observed during the sintering of this APC residue at 500 degrees C in a rotary furnace. The 168 h leaching test on filter residue, representing the fraction that may escape the flue gas treatment system, indicated rapid release of Pb and other contaminants. Caracolite and KCl x 2PbCl2 are significantly dissolved, and anglesite and cotunnite form the alteration products, as was confirmed by mineralogical analysis and PHREEQC-2 modeling. The observed Pb-bearing chlorides have significantly higher solubility than anglesite and, following emission from the smelter stack, can readily dissolve, transferring Pb into the environmental milieu (soils, water, inhabited areas). PMID- 16382958 TI - Push-pull tests to quantify in situ degradation rates at a phytoremediation site. AB - Nine push-pull tests (PPTs) were performed to determine in-situ aerobic respiration rates at a creosote-contaminated site and to assess the contribution of hybrid poplar trees to the remediation of polynuclear aromatic hydrocarbons (PAH) in groundwater. PPTs were conducted by injecting a solution containing dissolved oxygen and naphthalene (reactive tracers) with bromide (nonreactive tracer) into wells constructed in a shallow unconfined aquifer. The objective of this study was to determine seasonal variation and spatial differences (contaminated versus uncontaminated areas and treed versus untreed areas) in the rate of consumption of dissolved oxygen. First-order aerobic respiration rates varied from 0.0 (control well) to 1.25 hr(-1), which occurred at a planted area in early summer (June). Rates measured in winter at treed areas were greater by a factor of 3-5 when compared to winter rates determined at nontreed areas of the site. Rates at treed regions were found to increase by over 4 times in summer relative to winter at the same location. PMID- 16382959 TI - Improvement of the desulfurization and regeneration properties through the control of pore structures of the Zn-Ti-based H2S removal sorbents. AB - To improve the sulfur removing capacity of the conventional Zn-Ti-based H2S removal sorbents, a new Zn-Ti based sorbent (ZT-cp) was prepared by the coprecipitation method and tested in a packed bed reactor at middle temperature conditions (H2S absorption at 480 degrees C, regeneration at 580 degrees C). The new Zn-Ti-based sorbent showed excellent sulfur removing capacity without deactivation, even after 10 cycles of absorption and regeneration. The conventional Zn-Ti-based sorbents (ZT-700, ZT-1000), however, that were prepared by physical mixing, were continuously deactivated. In particular, the initial sulfur removing capacity of the ZT-cp sorbent showed a very high absorption value (0.22 g S/g sorbent), which corresponded to 91.6% of theoretical absorption amount. These results can be explained by the difference in physical properties such as pore volume, surface area, and particle size. It was also found that the sulfides formed from the ZT-cp and ZT-1000 sorbents with spinel structure were easily regenerated even at 580 degrees C. Those from the ZT-700 sorbent, with separated ZnO and TiO2 structures, needed a temperature higher than 610 degrees C for regeneration. PMID- 16382960 TI - Determination of kinetic law for toxic metals release during thermal treatment of model waste in a fluid-bed reactor. AB - Accumulation of toxic metals generated by thermal treatment of municipal solid waste presents a serious threat to the environment. A study was carried out to investigate the kinetic law of toxic metal release from municipal solid waste during their thermal treatment. Both direct and inverse models were developed in transient conditions. The direct mathematical model of the fluid-bed reactor is based on Kunii and Levenspiel's two-phase flow model for Geldart Group B particles. The inverse model intends to predict the metal's rate of vaporization from its concentration in the outlet gas. The derived models were found to predict reasonably well the experimental observations. A method to derive the kinetic law of toxic metals release during fluidized bed thermal treatment of model waste from the global model and the experimental measurements is derived and illustrated. A first-order law was fitted for the mineral matrix, and a second-order law (simplified) was fitted for the realistic model waste. The kinetic law obtained in this way could be integrated in a global model of combustion of municipal solid waste in order to simulate the effects of operating parameters on the metal's behavior. PMID- 16382961 TI - Effect of hydroxypropyl-beta-cyclodextrin on the degradation of pentachlorophenol by potassium monopersulfate catalyzed with iron(III)-porphyrin complex. AB - A novel biomimetic catalytic system containing a supramolecular complex between iron(III)-tetrakis(p-sulfonatophenyl)porphyrin [Fe(III)-TPPS] and hydroxypropyl beta-cyclodextrin (HP-beta-CD) was examined for the potassium monopersulfate catalyzed oxidation of pentachlorophenol (PCP). In the absence of HP-beta-CD, the percentage of PCP disappearance and the numbers of chlorine atoms released from PCP increased to 50% and 1.5 for a 1-day reaction period, respectively. However, in the presence of HP-beta-CD, the PCP completely disappeared and the number of chlorine atoms from PCP was increased to 3.1. o-Tetrachloroquinone, 2- and 4 hydroxyl-nonachlorodiphenyl ethers, and octachlorodibenzo-p-dioxin were detected among the oxidation products. In the absence of HP-beta-CD, the percentage of PCP conversion to oxidation products increased and then reached plateau. In the presence of HP-beta-CD, the amount of oxidation products produced initially increased for the first 10 min and thereafter decreased gradually. These results suggest that the addition of HP-beta-CD results in the further degradation of oxidation products. In addition, the mineralization of PCP to CO2 was investigated using 14C6-labeled PCP. After a 1-day reaction period, 24% of the 14C6-labeled PCP was converted to 14CO2 in the presence of HP-beta-CD, although significant 14CO2 generation was not observed in its absence. The effect of HP beta-CD on the facilitation of PCP degradation can be attributed to the fact that the self-oxidation of Fe(III)-TPPS is prevented by the formation of a stable supramolecular complex between HP-beta-CD and Fe(III)-TPPS. PMID- 16382962 TI - Modeling Cryptosporidium parvum oocyst inactivation and bromate formation in a full-scale ozone contactor. AB - The inactivation of Cryptosporidium parvum oocysts and the formation of bromate were assessed simultaneously by performing experiments with a full-scale ozone bubble-diffuser contactor used for drinking water disinfection. Fluorescence-dyed polystyrene microspheres were used as surrogates for C. parvum oocysts. Semi batch ozonation experiments were performed to determine the fluorescence intensity decay of individual microspheres, which was measured by flow cytometry. The results obtained with the microspheres were correlated to the inactivation kinetics of C. parvum oocysts by choosing an appropriate threshold fluorescence intensity below which microspheres were considered to be equivalent to nonviable oocysts. A mathematical model was then used to predict the inactivation efficiency and bromate formation. The contactor hydrodynamics were characterized by running tracer tests, and the kinetic parameters for ozone decomposition and bromate formation were obtained by performing batch experiments. Model predictions were in good agreement with full-scale experimental results. Additional model simulations revealed that ozone contactors should be designed with the lowest possible backmixing so that the target inactivation efficiency can be achieved with the lowest possible formation of bromate. PMID- 16382963 TI - Inhibition of biohydrogen production by undissociated acetic and butyric acids. AB - Glucose fermentation to hydrogen results in the production of acetic and butyric acids. The inhibitory effect of these acids on hydrogen yield was examined by either adding these acids into the feed of continuous flow reactors (external acids), or by increasing glucose concentrations to increase the concentrations of acids produced by the bacteria (self-produced). Acids added to the feed at a concentration of 25 mM decreased H2 yields by 13% (acetic) and 22% (butyric), and 60 mM (pH 5.0) of either acid decreased H2 production by >93% (undissociated acid concentrations). H2 yields were constant at 2.0 +/- 0.2 mol H2/mol glucose for an influent glucose concentration of 10-30 g/L. At 40 g glucose/L, H2 yields decreased to 1.6 +/- 0.1 mol H2/mol glucose, and a switch to solventogenesis occurred. A total undissociated acid concentration of 19 mM (self-produced acids) was found to be a threshold concentration for significantly decreasing H2 yields and initiating solventogenesis. Hydrogen yields were inhibited more by self produced acids (produced at high glucose feed concentrations) than by similar concentrations of externally added acids (lower glucose feed concentrations). These results show the reason hydrogen yields can be maximized by using lower glucose feed concentrations is that the concentrations of self-produced volatile acids (particularly butyric acid) are minimized. PMID- 16382964 TI - Avian toxicity reference values for perfluorooctane sulfonate. AB - Toxicity reference values (TRVs) and predicted no effect concentrations (PNECs) were derived for perfluorooctane sulfonate (PFOS) based on the characteristics of a top avian predator. On the basis of the protective assumptions used in this assessment, the benchmarks are protective of avian populations and were based on acute and chronic dietary exposures of northern bobwhite quail and mallard. Toxicological endpoints included mortality, growth, feed consumption, and histopathology. Reproductive endpoints included egg production, fertility, hatchability and survival, and growth of offspring. On the basis of the U. S. Environmental Protection Agency Great Lakes Initiative methodology, and a lowest observable adverse effect concentration (LOAEC) of 10 mg PFOS kg(-1) feed, an uncertainty factor of 36 was derived. The TRV based on PFOS dietary intake was 0.021 mg PFOS kg(-1) body weight day(-1), while for serum, liver, and egg, TRVs were 1.7 microg PFOS mL(-1), 0.6 microg PFOS g(-1) wet weight, and 1.7 microg PFOS mL(-1), respectively. On the basis of the European Commission methodology, a correction factor of 2 (for lowest observed effect level to no observable effect level) and an assessment factor of 30, for a total adjustment of 60, were used to derive PNECs. PNECs based on dietary, mean serum, liver, and egg PFOS concentrations were 0.013 mg PFOS kg(-1) body weight day(-1), 1.0 microg PFOS mL( 1), 0.35 microg PFOS g(-1) wet weight, and 1.0 microg PFOS mL(-1), respectively. PMID- 16382965 TI - Uptake of aqueous and dietary metals by mussel Perna viridis with different Cd exposure histories. AB - The influences of different Cd pre-exposure regimes (route, concentration, and duration of Cd exposure) on the bioavailability of Cd, Ag, Hg, and Zn to the green mussels Perna viridis were quantified in this study. Following pre-exposing the mussels to Cd, we measured the mussel's tissue Cd concentration and clearance rate, as well as the metal dietary assimilation efficiency (AE) and the influx rate from the dissolved phase of the four studied metals. Differences in the route (aqueous and dietary pathways) and the history of pre-exposure (combined Cd concentration and duration) did not significantly affect the subsequent Cd dietary and aqueous uptake. The Cd dietary AEs increased following both the dissolved and dietary Cd pre-exposure. There was a significant correlation between the Cd AE and the accumulated Cd body concentration in the mussels. Dietary assimilation of Hg and Zn also increased slightly (but not significantly) after Cd pre-exposure, but the AEs of Ag remained constant. Except for the significant decrease in the dissolved uptake of Hg, Cd pre-exposure did not apparently affect the uptake of the other three metals from the solution. Metal metal interactions are likely to be affected by the specificity of metallothionein induction. Our study demonstrated that the Cd body concentration as well as the environmental Cd concentration instead of the history of pre exposure was more important in affecting the Cd accumulation in the mussels. Such factors need to be considered in interpreting metal body concentrations in biomonitors. PMID- 16382966 TI - Oxide nanoparticle uptake in human lung fibroblasts: effects of particle size, agglomeration, and diffusion at low concentrations. AB - Quantitative studies on the uptake of nanoparticles into biological systems should consider simultaneous agglomeration, sedimentation, and diffusion at physiologically relevant concentrations to assess the corresponding risks of nanomaterials to human health. In this paper, the transport and uptake of industrially important cerium oxide nanoparticles, into human lung fibroblasts is measured in vitro after exposing thoroughly characterized particle suspensions to a fibroblast cell culture for particles of four separate size fractions and concentrations ranging from 100 ng g(-1) to 100 microg g(-1) of fluid (100 ppb to 100 ppm). The unexpected findings at such low but physiologically relevant concentrations reveal a strong dependence of the amount of incorporated ceria on particle size, while nanoparticle number density or total particle surface area are of minor importance. These findings can be explained on the basis of a purely physical model. The rapid formation of agglomerates in the liquid is strongly favored for small particles due to a high number density while larger ones stay mainly unagglomerated. Diffusion (size fraction 25-50 nm) or sedimentation (size fraction 250-500 nm) limits the transport of nanoparticles to the fibroblast cells. The biological uptake processes on the surface of the cell are faster than the physical transport to the cell at such low concentrations. Comparison of the colloid stability of a series of oxide nanoparticles reveals that untreated oxide suspensions rapidly agglomerate in biological fluids and allows the conclusion thatthe presented transport and uptake kinetics at low concentrations may be extended to other industrially relevant materials. PMID- 16382968 TI - [In vitro and in vivo biopharmaceutical evaluation of lorazepam commercial tablets]. AB - OBJECTIVE: Lorazepam (LZM) is a broadly used tranquillizer for the treatment of anxiety. Tavor is one of the most diffused registered drug products containing LZM. Lots of generic drug products containing LZM have been registered in Italy by several pharmaceutical groups and are present in the Italian market. Due to the wide medical prescription of products containing LZM, it seemed interesting, from a technological and biopharmaceutical point of view, to perform a comparative bioequivalence evaluation of a trade marked and generic tablet formulations containing LZM available in the Italian market. MATERIALS AND METHODS: The trial vas carried out on four preparations, including Tavor (1 mg) as reference product and three generics named A, B and C. In vitro technological parameters (dissolution, uniformity of content, uniformity of weight) and in vivo (pharmacokinetic on rabbit) studies were performed. RESULTS: All the examined brands passed technological tests according the European Pharmacopeia 5th ed. and USP25. Tavor ensures a faster dissolution behaviour than the tree generics in vitro. All the pharmacokinetic parameters were within the prescriptions of EMEA, even if the reference product showed the highest values of AUC0-infinity and of Cmax. CONCLUSIONS: Obtained results show that Tavor ensures a faster dissolution behaviour than the tree generics in vitro. Moreover, considering the pharmacokinetic data, it is possible to suppose that the generics B and C are not able to provide the same therapeutic effect as the reference product. PMID- 16382967 TI - Surgical management of substernal goitres. When is sternotomy inevitable? AB - PURPOSE: Aim of this retrospective study is to report personal experience in the surgical management of substernal goitres emphasizing the guidelines for preoperative planning of sternotomy in selected cases. PATIENTS AND METHODS: Medical records of all patients (n=355) submitted to thyroidectomy for struma in our Operative Unit, between 1993-2003, were analysed. A substernal goitre was defined as a goitre having a significant retrosternal extension (>50%) requiring mediastinal dissection. RESULTS: A total of 18 out of 355 patients undergoing thyroidectomy for struma in our Operative Unit had substernal goitres. The most common symptoms, at presentation, were the presence of neck mass and respiratory disorders. Standard cervical incision was adequate to achieve total thyroidectomy in 17 cases while, in one patient with computed tomography images showing the presence of a huge goitre extending below the aortic arch, a sternotomic approach was inevitable to ensure safe removal. No major morbidity or perioperative deaths occurred. One patient with scleroderma experienced bilateral paralysis of laryngeal nerves for two months, with full recovery thereafter. CONCLUSIONS: While removal of the majority of substernal goitres can be achieved by means of cervical incision, this approach is not always safe. In a selected number of cases with an iceberg shaped substernal goiter and with >70% of the volume lying below the thoracic outlet, a sternotomic approach is inevitable. Preoperative diagnostic work-up should, thus, include chest X-ray and computed tomography. Overall results in the present patient population, have been excellent since morbidity has been minimal and mortality absent, and all patients are symptom free. PMID- 16382969 TI - [Efficacy of adjunctive donepezil for cognitive impairments in patients with schizophrenia]. AB - Schizophrenia generally manifests cognitive disorders of subjects affected by this illness. Acetylcholine is the main neurotrasmettitor involved in the modulation of cognitive processes as attention, memory and executive functions. The aim of our study is to examine the effects of anticholinesterasic drugs in addiction to atypical antipsychotics on cognitive functions in subjects with schizophrenia. Participant to the study 14 subjects affected by schizophrenia. Subjects have been divided in two sub-groups on the grounds of pharmachological treatment used. A first group (N= 8) it's been treated only with risperidone (monotherapy group); the second one (N=7) it's been treated with donepezil in addition to risperidone (donepezil+ risperidone group). The group treated with donepezil + risperidone evidenced, after 3 and 6 months, statistically significant improvements in attention, in executive functions and in understanding first order Theory of Mind. Our findings are in agreement with those reported by MacEwan et al. (2001). Even if preliminary, our results prove the effectiveness of using anticholinesterasics drugs in addition to atypical antipsychotic treatment, especially in improving attentive functioning. PMID- 16382970 TI - [Primary hyperparathyroidism]. AB - Primary hyperparathyroidism (PHPT) is characterized by excessive PTH secretion in respect to calcium homeostasis needs, due to parathyroid adenoma (80% of cases), hyperplasia (15-20%), or carcinoma (1-2%). In familial forms of PHPT, several mutations have an established role: menin gene for MEN type 1, RET for MEN type 2a, calcium-sensing receptor gene for familial hypocalciuric hypercalcemia, parafibromin gene for PHPT-jaw tumour and carcinoma. Etiology of sporadic adenomas (80% of PHPT cases) is less defined, being most commonly found a mutation of menin gene or activation of PRAD1 oncogene. In recent years, the classical features of the disease became less common. Typically, bone involvement is now represented by a reduced bone mass at skeletal sites more rich in cortical tissue. Prominently trabecular skeletal sites are relatively spared, because of the anabolic effects of a slight PTH excess on trabecular tissue. PHPT patients may have increased fracture risk, though it is not clear why bone damage is more severe in a subgroup of patients. Clinical features of hypercalcemia may be fatigue, anorexia, thirst, and polyuria. Vague neurological and psychiatric symptoms, such as weakness, anxiety, depression, paresthesias, and muscular cramps may ameliorate after parathyroidectomy. Recent reports indicate increased cardiovascular mortality in PHPT patients. Diagnosis is based on the detection of hypercalcemia, together with inappropriately high serum PTH levels. Preoperative localization of the diseased glands is mandatory in persistent or recurrent PHPT, as like as when minimally invasive surgery is planned. High resolution ultrasonography and SPECT double-phase 99m Tc-sestamibi scintigraphy are the most commonly employed techniques. Intraoperatory PTH assay may confirm successful surgery when serum concentrations decrease more than 50%. Surgical therapy is indicated in patients with renal or skeletal complications, such as in those with previous parathyrotoxic crisis. Many surgeons in recent years adopted minimally invasive parathyroidectomy. Medical treatment is an option for patients unwilling or unfitted for surgery because of severe concomitant diseases. Employed therapy includes estrogens, SERMs, bisphosphonates and calcimimetics. PMID- 16382971 TI - [Nutritional problems in the surgical patients with head and neck tumours. Literature review and personal experience]. AB - During the last years the importance of the correct nutritional assessment as a part of the therapeutic process of the human pathologies has a greatly increased relevance. Still more in oncology, such relationship among nutritional assessment and good result of the therapeutic treatment have a fundamental importance. Among the neoplastic diseases those of the head and the neck result to be particularly aggressive, both for their nature and for the particular anatomical organization of such body areas, and the affected patients also need a precise evaluation of the nutritional assessment to obtain an optimal therapeutic result. On the basis of such premises and to their own clinical experience, the authors describe the main nutritional problems of the patients with tumours of the head and the neck and the possible therapeutic solutions. PMID- 16382972 TI - [Spa therapy in urolithiasis]. AB - In the present paper we remind epidemiology, etiopathogenesis and physiopathology of urolythiasis, emphasizing the role of lythogenic and antilythogenic urinary costituents. Mineral waters used in prevention and therapy of urolythiasis are described, namely oligomineral waters and, in uric lythiasis, bicarbonate mineral waters. We stress the activity of oligomineral waters, regarding their very low concentration of solute, presence of oligoelements, and antilythogenic components. At the end, we outline the role of spa therapy in the correction of metabolic disorders, which are etiologic and pathogenetic factors of urolythiasis. PMID- 16382973 TI - [Metabolic response to food and diet in paediatric obesity]. AB - Obesity has reached today epidemic proportions in industrialized countries with negative effects on children and adult's health, and excessive costs for society. Although genetics and ambient are surely implicated in determining the positive energy balance in the organism, to our known few has been written on the role of macronutrient (proteins, fats and carbohydrates) in diet, particularly in children, in the onset and develop of obesity. Children's eating patterns and habitudes have been hanged in the last years, so that there is an higher intake of PUFA (poliinsatured fat acids) and lower of MFA (monoinsatured fat acids), and an elevated density of carbohydrate in the diet. Considering the effects of macronutrients on body energetic metabolism , it is clear that in obese children there is an higher oxidation of exogenous carbohydrates and a lower oxidation of endogenous ones; this effect conducts in short term to an increment of tissue fat and to a consequent increment of body weight. High fat intake and fat oxidation influence the accumulation of fat mass rather over a long term in children. Fat oxidation is favourite also by the assumption of high glycemic index foods, that conducts to insulin-resistance. A low fat diet, rich in low glycemic index carbohydrates that can reduce hungry and avoid insulin resistance, acts together to a regular and aerobic constant physical activity, which helps fat mobilization from adipose tissue and from muscles, and can reduce insulin-resistance, so favouring weight loss. PMID- 16382974 TI - [Topical notes about work diseases rules]. AB - The Authors explain the text of the Decree April 27 2004 related to the new list of work diseases to be reported as by law enacted, instead of the previous Decree April 18 1973. The Decree follows up the latest UE Recommendation 2003/670 in order to bring up to date the European list of work diseases. The Authors examine several topical points of this decree, wishing for Italy a new up to date list for work diseases as from the last DPR April 13, 1994 no 336. PMID- 16382975 TI - [Terri's case and the ethical challenge for an always more technologically advanced medicine]. AB - BACKGROUND: Highly technological medicine puts the physician in the condition of verifying always more and always better the patients' vital parameters. It also opens up new issues on the ethical side because it makes more difficult placing the borderline between life and death. The relation between the guarantee of the necessary cures, aggressive treatment and a possible request for euthanasia often appears smooth in the experts perception and in the public opinion. Indeed it feels involved in strongly expressing its opinion in a field which can affect each one of us or else in the treatment relation with patients and relatives. THE CASE: Terri Schiavo's recent death, after fifteen year of vegetative coma, occurred as a consequence of the withdrawal of feeding-tubes, has spread from the USA to Italy and the rest of the world the dramatic difficulty of clarifying issue of this impact. On one side there are solutions of an emotional kind, based on pietas, these affect the relatives who caring for such patients. This first considerations reveals its contradiction in the fact that the parents wish to keep on yielding their daughter's life and thus clash with her husband who, on the other hand, wants to interrupt all kind of treatment. On the other hand there are serious issues of a clinical and ethical kind according to which normal treatment is guaranteed at all cost and nutrition has to be considered as ordinary treatment. Aggressive treatment can reach other not lawful dimensions because they are against human dignity and end up in worsening pain and suffering. CONCLUSIONS: Nowadays there is a pressing need for a wide information campaign for public opinion on these issues as well, as there is the possibility that all citizen can write a living will fully conscious of his responsibility. Furthermore it is even more urgent that medical class and all those who take care of these patients are able to harmonise their technical and scientific culture, keeping in mind a ethical and anthropological vision in order to fully understand the dignity of the dying during their last days, in an atmosphere of warm welcome and paying attention not only to the patients but to their close relatives as well. This because the weight of the treatment is on them, as is the psychological and emotional tension. The new kinds of burnout are so important that are being considered by present medicine as a new challenge. PMID- 16382976 TI - [Breast cancer: HER2 changes one's cards on the table]. AB - Recently, preliminary results of several randomized studies using trastuzumab in the adjuvant or neoadjuvant treatment of HER2-positive breast cancer have been reported. In the neoadjuvant setting, patients have been randomized to receive either chemotherapy alone (Group I), 4 cycles of paclitaxel followed by FEC (fluorouracil, epirubicin, cyclophosphamide) fo 4 cycles, or the same chemotherapy with concomitant weekly trastuzumab for 24 weeks (Group II). Pathologic complete responses were 25% in Group I and 66.7% in Group II, showing a significant superiority of treatment including trastuzumab. Among several ongoing studies of adjuvant therapy with trastuzumab, NSABP B-31 trial and NCCTG N9831 trial compared a standard treatment of sequential AC (doxorubicin, cyclophosphamide) followed by paclitaxel to the same chemotherapy regimen in combination with weekly trastuzumab for 1 year. In a third study (HERA trial), patients were randomized to 3 arms following adjuvant chemotherapy: observation, triweekly trastuzumab for 1 year or for 2 years. Joint analysis of B-31 and N9831 trials and interim analysis of patients randomized to receive 1 year trastuzumab in the HERA trial, show a significant improvement in disease-free survival with chemotherapy combined with trastuzumab. Treatment has been generally well tolerated with acceptable cardiotoxic effects (< 4%). However, the short follow up precludes any information about long-term side-effects. Overall, although the risk/benefit ratio is in favor of trastuzumab including regimens, the use of this monoclonal antibody in the neoadjuvant or adjuvant treatment of HER2-positive breast cancer, should be carefully discussed with the patient. PMID- 16382977 TI - [Epidemiology of malignant penile tumors]. PMID- 16382979 TI - [Anatomo-pathology of malignant penile tumors]. PMID- 16382978 TI - [Anatomic review of the penis]. PMID- 16382980 TI - [Anatomo-pathologic report and classifications of penile carcinomas]. PMID- 16382981 TI - [Recommendations: diagnosis of malignant penile tumors]. PMID- 16382982 TI - [Primary penile tumor: local treatment]. PMID- 16382983 TI - [Management of lymph node regions in malignant penile tumors]. PMID- 16382984 TI - [Chemotherapy of penile carcinomas]. PMID- 16382986 TI - [What is your diagnosis? Extradural brain meningioma with rhabdoid features]. PMID- 16382985 TI - [Pulmonary veins and atrial fibrillation: a pathological study of 100 hearts]. AB - Pathogenesis of atrial fibrillation (AF), the most common sustained heart arrhythmia, is not yet fully elucidated. Recent electrophysiological studies have shown that in most patients with AF the arrhythmia is triggered by ectopic beats originating from extensions of left atrial myocardium over the pulmonary veins (PVs), so called myocardial sleeves (MSPV). A total of 100 hearts (393 PVs) obtained at autopsy were prospectively studied - 50 from patients with chronic AF (average age 76.9 +/- 7.3 yrs.) and a control group of 50 with a sinus rhythm (aver. age 71.7 +/- 9.5 yrs.). This is a largest study published on this topic so far. It appeared that MSPV frequently harbour pathological lesions, particularly senile atrial amyloid, and scarring. These two pathological changes were evaluated semiquantitatively on a grade 0-3 basis in individual PVs, comparing the results in the AF vs. the control group. Amyloidosis of MSPV was found in 68 % of all hearts and in 55 % of all sleeves. The deposits were most marked in the right superior PV. Amyloidosis was more frequent and more severe in MSPV of patients with AF (58.5 %; average grade 0.89) than of those without AF (51.7 %; aver. grade 0.76); the differences, however, lack statistical significance. Scarring of MSPV was present in all 349 sleeves, more markedly in the left inferior, left superior, and right superior PVs. It was significantly more severe in patients with AF compared to those without the arrhythmia. By an injection metod, we have shown that MSPV are supplied by coronary arteries. However, the degree of scarring of the sleeves did not correlate with the degree of coronary atherosclerosis. We suggest that genesis of the scarring is not postnecrotic but degenerative, due to diffuse hypoxia of the sleeve myocardium. To conclude, amyloidosis and particularly scarring of MSPV appear generally in the elderly population as an arrhythmogenic substrate for AF. PMID- 16382987 TI - Desmin-positive and alpha-smooth muscle actin positive chondrocytes in human defective articular cartilage--preliminary report. AB - We studied the results of immunostaining for S-100 protein, alpha-smooth muscle actin, muscle specific actin and desmin in articular cartilage specimens obtained during an arthroscopy from eight patients with different degrees of osteoarthritis of the knee joint. In all cases, most of the cartilage cells were strongly positive for S-100 protein. Actin positive chondrocytes were present in four samples showing repair cartilage changes with occurrence of fibrocartilage tissue. Moreover, in one case, we observed typical desmin-positive chondrocytes in the layer of cartilage filling the defect of the articular cartilage surface. The expression of desmin can be regarded as a reaction to trauma or the indication of an inherent abnormality. The chondrocytes probably switched on smooth muscle features during the healing process, because desmin is to a great extent a typical muscle cell marker. This fact could probably support our previous supposition that cartilage cells expressing muscle markers could be designated as myochondroblasts and myochondrocytes analogously to the terminology of myofibroblasts. It is possible that during the healing of the cartilage defects, such a transformation of the immunophenotype of the cartilage cells is quite frequent, but it could also be only transient nature only. PMID- 16382988 TI - Autoimmune gastritis. A clinicopathologic study of 25 cases. AB - The histopathological diagnosis of autoimmune gastritis (AG) in its early stages can be a diagnostic challenge. Even some advanced cases with complete atrophy of the corpus mucosa may be difficult to recognize. To establish the diagnosis of autoimmune gastritis, several histological features should be assessed and combined with immunostains for enterochromaffin cell-like (ECL) cells and G cells. The main histological criteria include a mononuclear infiltrate within the lamina propria, foci of destruction of oxyntic glands, intestinal metaplasia (IM), pyloric metaplasia, and parietal cell pseudohypertrophy. These criteria were evaluated in our series of 25 patients with achlorhydria and/or megaloblastic anemia. Some of our patients presented with nonspecific gastrointestinal symptoms. The age ranged between 46 and 79 years; one male patient was only 31 years old. Histologically, the corpus mucosa displayed in all cases chronic inflammation with focal complete IM and advanced pyloric metaplasia. In 4 patients, oxyntic glands were destructed in some sites. There was a pancreatic metaplasia of acinar type in 2 patients and a minimal focal pseudohypertrophy of parietal cells in the 31-year-old man. A tubular adenoma with a low-grade dysplasia was found in one female patient. Immunohistochemically, chromogranin-A highlighted linear or nodular hyperplasia of ECL cells in 19 patients, and adenomatoid ECL hyperplasia in one case (80%). In the remaining cases hyperplasia of ECL cells could not be recognized from their normal count. In 13 cases (52%) a few ECL cells were seen also in IM. Regarding associated pathology, in one woman with nodular ECL cell hyperplasia, a gastric carcinoid was removed endoscopically. The reaction with gastrin antibody revealed in 11 cases (44%) a small number of G cells in IM in the corpus mucosa. In 18 patients, antral mucosa was examined as well. In 8 patients, the mucosa was normal; in 10 cases, a mild chronic inactive gastritis was diagnosed, and in 15 patients G-cell hyperplasia was found. In accordance with other studies, we show that the diagnosis of AG may be established microscopically in endoscopic specimens of the gastric body mucosa when histologic features and immunohistochemical detection of ECL and G cell hyperplasia are combined. PMID- 16382989 TI - [The role of intermedial filament nestin in malignant melanoma progression]. AB - Nestin is one of intermedial filaments exprimed in proliferating progenitor cells of the CNS and PNS (central and peripheral nervous system). Postnatal reexpression of the protein occures mainly in CNS tumors and correlates with a high grade of malignancy. The aim of our study is assessment of the nestin expression in benign and malignant skin melanocytic lesions with respect to presume a prognostic role of this protein. We examined 127 bioptic specimens, including 42 nodular melanomas (NM), 32 superficial spreading melanomas (SSM), 10 dysplastic nevi and 43 common intradermal or dermoepidermal nevi. We proved significant increase in nestin expression in melanoma groups, especially in nodular melanomas, where nestin was localized mainly in the peripheral, invasive areas of the tumor mass. CONCLUSION: Detection of nestin expression might be used as an additional melanocytic tumour marker. PMID- 16382990 TI - [Melanoma simulating malignant soft tissue tumour]. AB - The authors presented the case of an 82-year-old man with primary nodular melanoma of the skin on the back (Breslow 3 mm) which repeatedly metastasized five times to the cervical lymph nodes. Metastases were excised. The aim of this report was to demonstrate changes in the phenotype and immunophenotype during tumour progression. Originally round and oval melanoblasts had a characteristic immunophenotype. They were S100 protein, HMB45, Malan A, MITF positive. From the second biopsy the immunophenotype began to change, and the amount of positive cells declined. In the succeeding biopsies the morphology was also changed. There were spindle cells which formed mutually intermingled bundles in places with large multinucleate cells. The histological pattern assembled malignant mesenchymal tumour - type malignant fibrous histiocytoma. The immunophenotype was also changed. The second to fifth metastases in the cervical region were only S100 protein positive. The other above-mentioned melanoma markers were negative. Differential diagnosis includes neurotropic-desmoplastic malignant melanoma, Bednar tumour (pigmented dermatofibrosarcoma protuberans; storiform neurofibroma), malignant peripheral nerve sheath tumour (neurogenic sarcoma) and malignant fibrous histiocytoma. PMID- 16382991 TI - Gastrointestinal stromal tumor (GIST) with glandular component. A report of an unusual tumor resembling adenosarcoma. AB - A case of gastrointestinal stromal tumor (GIST) with an unusual glandular component is reported. The tumor was found in the gastric fundus of a 93-year-old woman. Histologically, the lesion showed a biphasic adenosarcoma-like structure. Typical low-grade spindle cell patterns of GIST were intermingled with numerous and partly cystic glands. The glandular epithelium had pyloric/foveolar-like appearance, with foci of intestinal metaplasia and low-grade dysplasia. The stromal component was immunoreactive for CD117 (c-kit) and CD34, and negative for myoid and neuroid markers. The ultrastructural examination found nondescript and undifferentiated spindle cells. The gastric mucosa and submucosa near the tumor contained a small area with features of gastritis cystica profunda, with glands similar to those present inside the tumor. Therefore, a collision of GIST and gastritis cystica profunda is suggested in the histogenesis of the lesion. PMID- 16382992 TI - Adenoid basal epithelioma of the uterine cervix in 21-year-old patient. Report of a case with histologic and immunohistochemical study. AB - A case of adenoid basal epithelioma (ABE) of the uterine cervix in 21-year-old woman is reported. The patient's age appears to be lowest among cases of ABE reported to date. The tumor showed typical histologic features of ABE and was associated with CIN3. In addition, rare tumor cells had surface cilia-appearing structures suggesting tubal differentiation. Immunohistochemically, the tumor was reactive for p63, CD10, ER, PR, p16 and bcl-2, and negative for CEA. Thus, the immunophenotype also suggests possible partial tuboendometrioid differentiation in the glandular component of the lesion. It further indicates, along with finding of associated CIN, a role of oncogenic HPV in pathogenesis of ABE. PMID- 16382993 TI - BK-virus nephropathy and simultaneous C4d positive staining in renal allografts. AB - The role of antibodies in rejection of transplanted kidneys was the subject of debate at the last two Banff meetings and in medical journals. Diffuse C4d positive staining of peritubular capillaries (PTCs) was recognized as a marker of antibody-mediated rejection and this morphological feature was included in the updated Banff schema. At the same time polyomavirus infection of the renal allografts has been reported more frequently and is emerging as an important cause of renal allograft dysfunction and graft loss. At the present time, BK virus nephropathy (BKN) represents the most common viral disease affecting renal allografts. BKN was identified in 6 patients in 12 biopsies and 2 graft nephrectomy specimens of 1115 biopsies between September 2000 and December 2003. Definite virus identification was done by immunohistochemistry. The reason for graft nephrectomies was graft failure due to BKN in a recipient after kidney pancreas transplantation with good function of his pancreas graft and the necessity of continuing immunosuppression. Detection of C4d deposits was performed by immunofluorescence or by immunohistochemistry. In graftectomy samples C4d detection was performed by immunohistochemistry and retrospectively in all cases of BKN. Focal C4d positive PTCs and BKN were found simultaneously in 9 of 12 needle biopsies and in both graft nephrectomy samples. Detection of C4d by immunohistochemistry disclosed focal C4d positive staining in kidney tissue but diffuse in the sites where BK-virus inclusions in tubular epithelial cells were found. The complement system is part of the host defense response and is crucial to our natural ability to ward off infection. In cases of BKN, virus likely gains access to the bloodstream through injured tubular walls and via PTCs. Vascular endothelium in the PTCs represents a potential target antigen for alloresponse, and simultaneously possibly represents an imprint of complement activation or complement production in the places with BK-virus infection. PMID- 16382995 TI - An overview of neuropeptide Y: pharmacology to molecular biology and receptor localization. PMID- 16382996 TI - Molecular ligand-receptor interaction of the NPY/PP peptide family. PMID- 16382997 TI - Modulator role of neuropeptide Y in human vascular sympathetic neuroeffector junctions. AB - Reverse transcription polymerase chain reaction (RT-PCR) studies identified the mRNA coding for the Y1 and Y2 receptors in human mammary artery/vein and saphenous vein biopsies. Y1 receptors are expressed in vascular smooth muscles and potentiate the contractile action of sympathetic co-transmitters, adenosine triphosphate (ATP) and noradrenaline (NA); BIBP 3226, a competitive Y1 receptor antagonist, blocked the neuropeptide Y (NPY)-induced modulation. The Y2 receptor is expressed in sympathetic nerves terminals and modulates the pool of sympathetic co-transmitters released at the neuroeffector junction. NPY plays a dual role as a modulator of sympathetic co-transmission; it facilitates vascular smooth muscle reactivity and modulates the presynaptic release of ATP and NA. Sympathetic reflexes regulate human vascular resistance, where NPY plays a modulator role of paramount importance following increased sympathetic discharges, such as stress and vascular disease. PMID- 16382998 TI - NPY and NPY receptors: presence, distribution and roles in the regulation of the endocardial endothelium and cardiac function. PMID- 16382999 TI - Neuropeptide Y and sympathetic control of vascular tone in hypertension. PMID- 16383000 TI - Neuropeptide Y and the cerebral circulation. PMID- 16383001 TI - Neuropeptide Y and the heart: implication for myocardial infarction and heart failure. PMID- 16383002 TI - Role of neuropeptide Y in the regulation of kidney function. AB - The presence in the mammalian kidney of NPY and at least one of its receptor subtypes has been proven by several independent methodologies. Also, numerous studies using physiological and pharmacological approaches indicated that this peptide has the capacity to alter renal function. In particular, these studies suggest that NPY may exert renal vasoconstrictor and tubular actions that are species dependent, and may also influence renin secretion by the kidney. The question whether NPY plays an important role in the physiological regulation of renal hemodynamics and electrolyte excretion, remains largely unanswered at present. No major impairments in renal function have been reported in genetically models deficient in NPY or its Y1 receptor. Thus, additional studies are required to elucidate the role of NPY in the physiological and pathophysiological regulation of renal function. PMID- 16383003 TI - NPY and neuron-adipocyte interactions in the regulation of metabolism. PMID- 16383004 TI - NPY and energy homeostasis: an opportunity for novel anti-obesity therapies. PMID- 16383005 TI - Subjugation of hypothalamic NPY and cohorts with central leptin gene therapy alleviates dyslipidemia, insulin resistance, and obesity for life-time. AB - An interactive network comprised of neuropeptide Y (NPY) and cohorts is obligatory in the hypothalamic integration of appetite and energy expenditure on a minute-to-minute basis. High or low abundance of NPY and cognate receptors dysregulates the homeostatic milieu engendering hyperphagia, decreased energy expenditure, obesity and attendant metabolic syndrome cluster of dyslipidemia, glucose intolerance, insulin resistance and hyperinsulinemia, risk factors for type II diabetes and cardiovascular diseases. Increasing the supply of the endogenous repressor hormone leptin locally in the hypothalamus with the aid of leptin gene therapy, blocked age-related and dietary obesities, and the sequential development of dyslipidemia, hyperglycemia, and insulin resistance. Thus, sustained repression of NPY signaling with increased leptin selectively in the hypothalamus can avert environmental obesity and the risks of metabolic diseases. PMID- 16383006 TI - NPY and bone. PMID- 16383007 TI - NPY in alcoholism and psychiatric disorders. PMID- 16383009 TI - NPY and extreme stress: lessons learned from posttraumatic stress disorder. PMID- 16383008 TI - Plasticity of neuropeptide Y in the dentate gyrus after seizures, and its relevance to seizure-induced neurogenesis. AB - In summary, NPY is clearly an important peptide in the adult rat dentate gyrus because it has the potential to influence synaptic transmission and neurogenesis. It may even have other functions, as yet undiscovered, mediated by glia or vasculature. The remarkable plasticity of NPY puts it in a position to allow dentate gyrus function to be modified in a changing environment. The importance of this plasticity in the context of epilepsy cannot be emphasized enough. It could help explain a range of observations about epilepsy that currently is poorly understood. For example, rapid increases in NPY could mediate postictal depression, the period of depression that can last for several hours after generalized seizures. It may mediate the "priming effect," which is a reduction in seizure threshold following an initial period of seizures. Finally, it could contribute to the resistance of dentate granule cells to degeneration after seizures. However, despite the focus in this review on seizure-induced changes, the changes described here also appear to occur after other types of manipulations, which considerably broadens the scope of NPY's role in the brain. PMID- 16383010 TI - NPY and chronic neurodegenerative disease. PMID- 16383011 TI - Human NPY gene variants in cardiovascular and metabolic diseases. PMID- 16383012 TI - Cancer morphology, carcinogenesis and genetic instability: a background. AB - Morphological abnormalities of both the nuclei and the cell bodies of tumour cells were described by Muller in the late 1830s. Abnormalities of mitoses and chromosomes in tumour cells were described in the late 1880s. Von Hansemann, in the 1890s, suggested that tumour cells develop from normal cells because of a tendency to mal-distribution and other changes of chromosomes occurring during mitosis. In the first decades of the 20th century, Mendelian genetics and "gene mapping" of chromosomes were established, and the dominant or recessive bases of the familial predispositions to certain tumour types were recognised. In the same period, the carcinogenic effects of ionising radiations, of certain chemicals and of particular viruses were described. A well-developed "somatic gene-mutational theory" of tumours was postulated by Bauer in 1928. In support of this, in the next three decades, many environmental agents were found to cause mitotic and chromosomal abnormalities in normal cells as well as mutations in germ-line cells of experimental animals. Nevertheless, mitotic, chromosomal, and other mutational theories were not popular explanations of tumour pathogenesis in the first half of the 20th century. Only in the 1960s did somatic mutational mechanisms come to dominate theories of tumour formation, especially as a result of the discoveries of the reactivity of carcinogens with DNA, and that the mutation responsible for xeroderma pigmentosum causes loss of function of a gene involved in the repair of DNA after damage by ultraviolet light (Cleaver in 1968). To explain the complexity of tumourous phenomena, "multi-hit" models gained popularity over "single-hit" models of somatic mutation, and "epigenetic" mechanisms of gene regulation began to be studied in tumour cells. More recently, the documentation of much larger-than-expected numbers of genomic events in tumour cells (by Stoler and co-workers, in 1999) has raised the issue of somatic genetic instability in tumour cells, a field which was pioneered in the 1970s mainly by Loeb. Here these discoveries are traced, beginning with "nuclear instability" though mitotic-and chromosomal theories, single somatic mutation theories, "multi-hit" somatic theories, "somatic, non-chromosomal, genetic instability" and epigenetic mechanisms in tumour cells as a background to the chapters which follow. PMID- 16383014 TI - Aneuploidy, stem cells and cancer. AB - Telomeres which protect the individual chromosomes from disintegration, end-to end fusion and maintain the genomic integrity during the somatic cell divisions play an important role in cellular aging. Aging and cancer development are linked with each other because cancer is considered a group of complex genetic diseases that develop in old cells and, in both, telomere attrition is involved. Numeric chromosome imbalance also known as aneuploidy is the hallmark of most solid tumors, whether spontaneous or induced by carcinogens. We provide evidence in support of the hypothesis that telomere attrition is the earliest genetic alteration responsible for the induction of aneuploidy. Dysfunctional telomeres are highly recombinogenic leading to the formation of dicentric chromosomes. During cell divisions, such complex chromosome alterations undergo breakage fusion bridge cycles and may lead to loss of heterozygosity (LOH) and gene amplification. Furthermore, we have provided evidence in support of the hypothesis that all types of cancer originate in the organ- or tissue-specific stem cells present in a particular organ. Cancer cells and stem cells share many characteristics, such as, self-renewal, migration, and differentiation. Metaphases with abnormal genetic constitution present in the lymphocytes of cancer patients and in some of their asymptomatic family members may have been derived from the organ-specific stem cells. In addition, evidence and discussion has been presented for the existence of cancer-specific stem cells. Successful treatment of cancer, therefore, should be directed towards these cancer stem cells. PMID- 16383013 TI - Abnormalities of chromatin in tumor cells. AB - Nuclear morphometric descriptors such as nuclear size, shape, DNA content and chromatin organization are used by pathologists as diagnostic markers for cancer. Tumorigenesis involves a series of poorly understood morphological changes that lead to the development of hyperplasia, dysplasia, in situ carcinoma, invasive carcinoma, and in many instances finally metastatic carcinoma. Nuclei from different stages of disease progression exhibit changes in shape and the reorganization of chromatin, which appears to correlate with malignancy. Multistep tumorigenesis is a process that results from alterations in the function of DNA. These alterations result from stable genetic changes, including those of tumor suppressor genes, oncogenes and DNA stability genes, and potentially reversible epigenetic changes, which are modifications in gene function without a change in the DNA sequence. DNA methylation and histone modifications are two epigenetic mechanisms that are altered in cancer cells. The impact of genetic (e.g., mutations in Rb and ras family) and epigenetic alterations with a focus on histone modifications on chromatin structure and function in cancer cells are reviewed here. PMID- 16383015 TI - The mode of action of organic carcinogens on cellular structures. AB - Most genotoxic organic carcinogens require metabolic activation to exert their detrimental effects. The present review summarizes the mechanisms of how organic carcinogens are bioactivated into DNA-reactive descendants. Beginning with the history of discovery of some important human organic carcinogens, the text guides through the development of the knowledge on their molecular mode of action that has grown over the past decades. Some of the most important molecular mechanisms in chemical carcinogenesis, the role of the enzymes involved in bioactivation, the target gene structures of some ultimate carcinogenic metabolites, and implications for human cancer risk assessment are discussed. PMID- 16383016 TI - Metal ions and carcinogenesis. AB - Metals are essential for the normal functioning of living organisms. Their uses in biological systems are varied, but are frequently associated with sites of critical protein function, such as zinc finger motifs and electron or oxygen carriers. These functions only require essential metals in minute amounts, hence they are termed trace metals. Other metals are, however, less beneficial, owing to their ability to promote a wide variety of deleterious health effects, including cancer. Metals such as arsenic, for example, can produce a variety of diseases ranging from keratosis of the palms and feet to cancers in multiple target organs. The nature and type of metal-induced pathologies appear to be dependent on the concentration, speciation, and length of exposure. Unfortunately, human contact with metals is an inescapable consequence of human life, with exposures occurring from both occupational and environmental sources. A uniform mechanism of action for all harmful metals is unlikely, if not implausible, given the diverse chemical properties of each metal. In this chapter we will review the mechanisms of carcinogenesis of arsenic, cadmium, chromium, and nickel, the four known carcinogenic metals that are best understood. The key areas of speciation, bioavailability, and mechanisms of action are discussed with particular reference to the role of metals in alteration of gene expression and maintenance of genomic integrity. PMID- 16383017 TI - Actions of ultraviolet light on cellular structures. AB - Solar radiation is the primary source of human exposure to ultraviolet (UV) radiation. Overexposure without suitable protection (i.e., sunscreen and clothing) has been implicated in mutagenesis and the onset of skin cancer. These effects are believed to be initiated by UV-mediated cellular damage, with proteins and DNA as primary targets due to a combination of their UV absorption characteristics and their abundance in cells. UV radiation can mediate damage via two different mechanisms: (a) direct absorption of the incident light by the cellular components, resulting in excited state formation and subsequent chemical reaction, and (b) photosensitization mechanisms, where the light is absorbed by endogenous (or exogenous) sensitizers that are excited to their triplet states. The excited photosensitizers can induce cellular damage by two mechanisms: (a) electron transfer and hydrogen abstraction processes to yield free radicals (Type I); or (b) energy transfer with O2 to yield the reactive excited state, singlet oxygen (Type II). Direct UV absorption by DNA leads to dimers of nucleic acid bases including cyclobutane pyrimidine species and pyrimidine (6-4) pyrimidone compounds, together with their Dewar isomers. These three classes of dimers are implicated in the mutagenicity of UV radiation, which is typified by a high level of CC-->TT and C-->T transversions. Single base modifications can also occur via sensitized reactions including Type 1 and Type II processes. The main DNA product generated by (1)O2 is 8-oxo-Gua; this is a common lesion in DNA and is formed by a range of other oxidants in addition to UV. The majority of UV-induced protein damage appears to be mediated by (1)O2, which reacts preferentially with Trp, His, Tyr, Met, Cys and cystine side chains. Direct photo-oxidation reactions (particularly with short-wavelength UV) and radicals can also be formed via triplet excited states of some of these side chains. The initial products of (1)O2-mediated reactions are endoperoxides with the aromatic residues, and zwitterions with the sulfur-containing residues. These intermediates undergo a variety of further reactions, which can result in radical formation and ring opening reactions; these result in significant yields of protein cross-links and aggregates, but little protein fragmentation. This review discusses the formation of these UV-induced modifications and their downstream consequences with particular reference to mutagenesis and alterations in protein structure and function. PMID- 16383018 TI - Actions of radiation on living cells in the "post-bystander" era. AB - Over the past 20 years there has been increasing evidence that cells and the progeny of cells surviving a dose of ionizing radiation can exhibit a wide range of effects inconsistent with the level of dose received. Recently, the cause of these delayed effects has been ascribed to so-called bystander effects, occurring in cells not directly hit by an ionizing track, but which are influenced by signals from irradiated cells. These effects are not necessarily deleterious, although most of the literature deals with adverse delayed effects. What is important to consider is what, if anything, these effects mean for what is still the central dogma of radiobiology and radiation protection, i.e., that DNA double strand breaks are the primary radiation-induced lesion that can be quantifiably related to received dose, and which determine the probability that a cancer will result from a radiation exposure. In this chapter we review the history of radiation biology which led to the DNA paradigm. We explore the issues and the evidence which are now challenging the view that dose deposition in DNA is all important. We conclude that in the low-dose region, the primary determinant of radiation exposure outcome is the genetic and epigenetic background of the individual and not the dose. This effectively dissociates dose from effect as a quantitative relationship, but it does not necessarily mean that the effect is unrelated to DNA damage somewhere in the system. PMID- 16383019 TI - Viral carcinogenesis and genomic instability. AB - Oncogenes encoded by human tumor viruses play integral roles in the viral conquest of the host cell by subverting crucial and relatively non-redundant regulatory circuits that regulate cellular proliferation, differentiation, apoptosis and life span. Human tumor virus oncoproteins can also disrupt pathways that are necessary for the maintenance of the integrity of host cellular genome. Some viral oncoproteins act as powerful mutator genes and their expression dramatically increases the incidence of host cell mutations with every round of cell division. Others subvert cellular safeguard mechanisms intended to eliminate cells that have acquired abnormalities that interfere with normal cell division. Viruses that encode such activities can contribute to initiation as well as progression of human cancers. PMID- 16383020 TI - Abnormalities of cell structures in tumors: apoptosis in tumors. AB - A conceptual shift has occurred in recent years from considering cancer as simply a disease of deregulated cell proliferation to a view that incorporates the aberrant control of apoptosis into the equation. Apoptosis is an organized, genetically programmed cell death process by which multicellular organisms specifically destroy, dismantle and dispose of cells. In cancer cells, this tightly controlled process is suppressed by genetic lesions, allowing cancer cells to survive beyond their normal life span even in hostile environments that are prone to hypoxia and lack many trophic factor supports. In the last two decades, cancer researchers have made great strides in our understanding of the underlying molecular mechanism of apoptosis in chemoresistance generation and tumorigenesis. This tremendous increase in our knowledge of apoptosis in tumors has greatly impacted our perspective on carcinogenesis. Key regulators of apoptosis such as members of the Inhibitors of Apoptosis family and Bcl-2 family have been shown to play a pivotal role in allowing most cancer cells to escape apoptosis. The identification of specific targets involved in the suppression of apoptosis in cancer cells has facilitated the design and development of therapeutic strategies based on rational molecular approaches that aim to modulate apoptotic pathways. Many promising apoptosis-dependent strategies have been translated into clinical trials in the continued assessment of regimens that can effectively eradicate cancers. PMID- 16383021 TI - Molecular regulation of tumor angiogenesis: mechanisms and therapeutic implications. AB - Angiogenesis, the process of new capillary formation from a pre-existing vessel plays an essential role in both embryonic and postnatal development, in the remodeling of various organ systems, and in several pathologies, particularly cancer. In the last 20 years of angiogenesis research, a variety of angiogenic regulators, both positive and negative, have been identified. The discovery of several anti-angiogenic factors has led to the development of novel cancer therapies based on targeting a tumor's vascular supply. A number of these new therapies are currently being tested in clinical trials in the U.S.A. and elsewhere. A major advance in the field of anti-angiogenic therapy occurred recently when the FDA approved Avastin (bevacizumab), the first solely anti angiogenesis therapy approved for treatment of human cancer. While it has long been appreciated that tumor growth and progression are dependent on angiogenesis, it is only recently that progress has been made in elucidating the molecular mechanisms that regulate the earliest stage in the angiogenic program, the angiogenic switch. This checkpoint is characterized by the transition of a dormant, avascular tumor into an active, vascular one. Anti-angiogenic therapies to date have essentially been designed to suppress the neovasculature in established tumors. However, identifying the mechanisms that cause a tumor to acquire an angiogenic phenotype may lead to the discovery of new therapeutic modalities and complementary diagnostics that could be used to block the angiogenic switch, thereby preventing subsequent tumor progression. In this chapter on the role of angiogenesis in cancer, we (1) provide an overview of the process of angiogenesis with special regard to the molecules and physiological conditions that regulate this process, (2) review recent studies describing the use of anti-angiogenic approaches in the treatment of a variety of human cancers, and (3) discuss the recent literature focused on the study of the molecules and molecular mechanisms that may be regulating the initiation of the angiogenic phenotype in tumors, and the clinical impact that this knowledge may have in the future. PMID- 16383022 TI - Genetic and environmental factors in hereditary predisposition to tumors: a conceptual overview. AB - Cancer is a heritable disorder of somatic cells. Carcinogenesis at the cellular level is like an opened Japanese fan, because initiated cells grow in several directions and tumors suggest the edge of the fan by having many gene abnormalities. We discuss here the primal force and gene networks (federal headship) in renal carcinogenesis. The Eker (Tsc2 mutant) rat model of hereditary renal carcinoma (RC) is an example of a Mendelian dominantly inherited predisposition to a specific cancer in an experimental animal. Recently, we discovered a new hereditary renal carcinoma in the rat in Japan, and the rat was named the "Nihon" rat. We suggest that its predisposing (Bhd) gene is a novel renal tumor suppressor gene. We present these unique models as part of the study of problems in carcinogenesis; e.g., multistep carcinogenesis, cancer prevention and the development of the therapeutic treatments that can be translated to human patients, as well as how environmental factors interact with cancer susceptibility gene(s). PMID- 16383023 TI - Effects of ionizing radiation on cellular structures, induced instability and carcinogenesis. AB - Ionizing radiation is perhaps the most extensively studied human carcinogen. There have been a number of epidemiological studies on human populations exposed to radiation for medical or occupational reasons, as a result of protracted environmental exposures due to radiation accidents, or after atomic bombings. As a result of these studies exposure to ionizing radiation has been unambiguously linked to cancer causation. While cancer induction is the primary concern and the most important somatic effect of exposure to ionizing radiation, potential health risks do not only involve neoplastic diseases but also somatic mutations that might contribute to birth defects and ocular maladies, and heritable mutations that might impact on disease risks in future generations. Consequantly it is important we understand the long-term health risks associated with exposure to ionizing radiation. PMID- 16383024 TI - Genetic instability in human tumors. AB - Genetic, or genomic, instability refers to a series of observed spontaneous genetic changes occurring at an accelerated rate in cell populations derived from the same ancestral precursor. This is far from a new finding, but is one that has increasingly gained more attention in the last decade due to its plausible role(s) in tumorigenesis. The majority of genetic alterations contributing to the malignant transformation are seen in growth regulatory genes, and in genes involved in cell cycle progression and arrest. Genomic instability may present itself through alterations in the length of short repeat stretches of coding and non-coding DNA, resulting in microsatellite instability. Tumors with such profiles are referred to as exhibiting a mutator phenotype, which is largely a consequence of inactivating mutations in DNA damage repair genes. Genomic instability may also, and most commonly, results from gross chromosomal changes, such as translocations or amplifications, which lead to chromosomal instability. Telomere length and telomerase activity, important in maintaining chromosomal structure and in regulating a normal cell's lifespan, have been shown to have a function in both suppressing and facilitating malignant transformation. In addition to such direct sequence and structural changes, gene silencing through the hypermethylation of promoter regions, or increased gene expression through the hypomethylation of such regions, together, form an alternative, epigenetic mechanism leading to instability. Emerging evidence also suggests that dietary and environmental agents can further modulate the contribution of genetic instability to tumorigenesis. Currently, there is still much debate over the distinct classes of genomic instability and their specific roles in the initiation of tumor formation, as well as in the progressive transition to a cancerous state. This review examines the various molecular mechanisms that result in this genomic instability and the potential contribution of the latter to human carcinogenesis. PMID- 16383025 TI - Molecular mechanisms of human carcinogenesis. AB - Intensive research efforts during the last several decades have increased our understanding of carcinogenesis, and have identified a genetic basis for the multi-step process of cancer development. Tumors grow through a process of clonal expansion driven by mutation. Several forms of molecular alteration have been described in human cancers, and these can be generally classified as chromosomal abnormalities and nucleotide sequence abnormalities. Most cancer cells display a phenotype characterized by genomic hypermutability, suggesting that genomic instability may precede the acquisition of transforming mutations in critical target genes. Reduced to its essence, cancer is a disease of abnormal gene expression, and these genetic abnormalities contribute to cancer pathogenesis through inactivation of negative mediators of cell proliferation (including tumor suppressor genes) and activation of positive mediators of cell proliferation (including proto-oncogenes). In several human tumor systems, specific genetic alterations have been shown to correlate with well-defined histopathological stages of tumor development and progression. Although the significance of mutations to the etiological mechanisms of tumor development has been debated, a causal role for such genetic lesions is now commonly accepted for most human cancers. Thus, genetic lesions represent an integral part of the processes of neoplastic transformation, tumorigenesis, and tumor progression, and as such represent potentially valuable markers for cancer detection and staging. PMID- 16383026 TI - Mechanisms of abnormal gene expression in tumor cells. AB - Epigenetic mechanisms are involved in critical nuclear processes such as transcriptional control, genome stability, replication and repair. Recent evidence suggests that changes in the epigenetic repertoire can drive tumorigenesis. This review examines the latest experimental evidence that questions the mechanisms underlying the consequence of epigenetic changes in gene regulation and cancer development. PMID- 16383027 TI - [Pondering over the fluid resuscitation during burn shock stage]. PMID- 16383028 TI - [Intensive research on the cellular and molecular mechanism of burn shock and ischemic and hypoxic injury]. PMID- 16383029 TI - [Study on the changes in the distribution and chemical states of the hepatic intra-and extra-cellular sodium ion in the rats with severe burns at early stage]. AB - OBJECTIVE: To investigate the changes in the distribution and chemical states of the hepatic intra- and extra-cellular sodium ion in the rats with severe burns, so as to provide guidance for fluid resuscitation at early postburn stage. METHODS: Nineteen adult male Sprague-Dawley (SD) rats were employed in the study and were randomly divided into control (n = 12) and burn (n = 7) groups. The changes in the longitudinal (T1) and transverse (T2) relaxation times of hepatic intra-cellular and extra-cellular sodium in the two groups were studied with 23Na NMR spectroscopy and a shift reagent. RESULTS: After infusion of the shift reagent,the extra-cellular sodium content in rat liver decreased by 17%, with obvious increase in fast T2 component (P < 0.01), indicating an increase in the fraction of Na+ binding sites in the extra-cellular space. The characteristics of relaxation of intra-cellular sodium remained unchanged despite a 57% increment in intra-cellular sodium content. CONCLUSION: The deficiency of sodium as a permeable molecule might be related to the postburn movement of hypertonic sodium from extra-cellular to intra-cellular space. The results indicated that it is reasonable to administer high concentration of sodium in fluid resuscitation during the first 24 postburn hours. PMID- 16383030 TI - [Protective effect of glycine on hypoxic rat myocardial cells]. AB - OBJECTIVE: To investigate the protective effect of glycine (Gly) on hypoxic rat myocardial cells and its mechanism. METHODS: Sdfetal rat myocardial cells were isolated and cultured in vitro. The released amounts of creatine kinase (CK) and lactate dehydrogenase (LDH) from the myocardial cells in the culture supernatant at 6 hour after hypoxia and after glycine treatment were determined with ultraviolet spectrophotometer. The expression of the alpha1 subunits of glycine receptor (GlyRalpha1) in the myocardial cells was detected by immunofluorescent histochemistry. The changes in the intracellular calcium content and the membrane potential of the myocardial cells were determined by laser confocal microscopy. RESULTS: The release of CK and LDH in the culture supernatant increased significantly at 6 h after hypoxia [(393.8 +/- 5.3), (1564 +/- 41) U/L] compared with those before hypoxia, while their levels were obviously decreased after glycine treatment [(56.3 +/- 2.7), (716 +/- 18) U/L, (P <0.01)] compared with those before glycine treatment. There was positive expression of GlyRalpha1 in myocardial cells before and after hypoxia. The average fluorescent intensity of intracellular calcium at 6 hours after hypoxia (139 +/- 29) was significantly higher than that before hypoxia (27 +/- 8, P < 0.01), while it was obviously lower (51 +/- 11) after glycine treatment compared with that at 6 hours after hypoxia,but it was evidently higher than that before hypoxia (P <0.01). The membrane potential 6 hours after hypoxia (62 +/- 9) was obviously lower than that before hypoxia (177 +/- 20, P < 0.01), but it was obviously higher after glycine treatment (123 +/- 16) than that at 6 hours after hypoxia (P < 0.01). CONCLUSION: Glycine might be beneficial in the protection of myocardial cells against hypoxia. The underlying mechanism may involve attenuation of membrane potential depolarization after hypoxia by conjugation of glycine with its receptor, depleting in turn voltage-dependent calcium channel on the cellular membrane, preventing calcium overload due to influx of calcium ions after hypoxia. PMID- 16383031 TI - [The influence of lipopolysaccharide on adipose metabolism in liver during shock stage of scalded rats]. AB - OBJECTIVE: To investigate the influence of lipopolysaccharide (LPS) on adipose metabolism in liver during shock stage of scalded rats. METHODS: Sixty adult Wistar rats were inflicted with 30% TBSA full thickness scald and were randomly divided into 3 groups: i. e. sham group (control, n = 20), simple scald group [(n = 20) and LPS group (n = 20, with intra-peritoneal injection of 3.0 mg/kg LPS at 2 postscald hour (PSH)]. The contents of LPS, tumor necrosis factor alpha (TNF alpha), free fatty acids (FFA) in plasma and adenosine triphosphate (ATP), triglyceride (TG), malonaldehyde (MDA) in liver in each group were determined at 24 and 48 PSH. The histological changes in hepatic tissue in each group were also observed. RESULTS: The plasma contents of FFA in LPS group at 24 and 48 PSH were 2.3 +/- 0.3 mmol/L and 2.5 +/- 0.4 mmol/L, respectively, which were obviously higher than those in control (0.4 +/- 0.3 mmol/L, 0.5 +/- 0.3 mmol/L) and scald (0.9 +/- 0.3, 1.2 +/- 0.5 mmol/L, P <0.01) groups. Meanwhile, there was obvious difference in the contents of TG and ATP in liver between LPS group (TG: 530 +/- 30 mmol/g, ATP: 1.7 +/- 0.5 micromol/g) and scald group (TG: 242 +/- 27 mmol/g, ATP: 6.0 +/- 2.4 micromol/g, P < 0.01). Pathological examination revealed that adipose denaturalization and injury to mitochondria in hepatocytes in scald group were significantly milder than those in LPS group. The morphology of hepatocyte in control group appeared normal. CONCLUSION: LPS challenge to burn subjects could induce impairment in utilizing fat derived energy, and it would aggravate adipose denaturalization in the liver. PMID- 16383032 TI - [The use of composite sodium lactate and sorbitol composition in fluid resuscitation for shock in patients with major burn]. AB - OBJECTIVE: To investigate the feasibility of the use of sodium lactate and sorbitol (CISS) in the fluid resuscitation for shock in patients with major burns. METHODS: Fifty - three adult patients with major burns (hospitalized within 6 hours after burns) were randomly divided into A (n = 24, with i.v. infusion of 50 g/L CISS, 2 000 ml per day) and B (n = 29, with i. v. infusion of 50 g/L glucose, 2 000 ml per day) groups. The amount of electrolytes and colloid as the main resuscitation fluids was calculated according to the formula in both groups. Meanwhile, additional electrolytes and insulin were supplemented to the patients in the B group. The result of combating shock, energy supply, and side effects in the two groups were observed. The changes in hepatic and renal function, and the changes in electrolytes were monitored. The amount of fluid supplementation and urinary volume were recorded. The level of blood glucose of each patient was determined at the admission time and 24, 48, and 72 hours after injury. RESULTS: No obvious difference was found in control of shock and energy supply between A and B group. There was no side effects or damage to hepatic and renal function related to infused fluids in A group. But the patients of the B group required supplementation of exra electrolytes and insulin during the fluid resuscitation period in order to maintain the normal levels of electrolytes and blood glucose, and this was not necessary in group A. The diuretic effect in group A was better than that in group B (average urinary volume in the first two 24 hours: group A: 1.9 +/- 0.6 and 3.3 +/- 0.8 L; group B:1.0 +/- 0.5 and 2.3 +/- 0.8 L). CONCLUSION: The use of CISS during shock stage of the patients with major burns could be beneficial to the replenishment of blood volume, control of shock, promotion of diuresis and subsidence of edema. It could also provide electrolytes and energy, without the influence on the level of blood glucose. PMID- 16383033 TI - [An experimental study on the influence of hypoxia induction factor-1alpha on the glycolysis of the rat myocardial cell under hypoxic condition]. AB - OBJECTIVE: To investigate the influence of hypoxia induction factor-1alpha (HIF 1alpha) on glycosis of rat myocardial cell under hypoxic condition. METHODS: The myocardial cells of the rats were routinely isolated and cultured. The cells were divided into single hypoxia (H) and HIF-1alpha inhibiting (I) groups. The cells in H group were cultured in glucose-free medium with mixed low-oxygen gas [1% O2, 94% N2 and 5% CO2 (v/v)]. While the cells in I group were cultured with low oxygen gas after the cell model of low expression of HIF-1alpha protein constructed by RNAi technique. The cells in both groups were all observed before hypoxia (routine culture) and at the time points of 1, 3, 6, 12 and 24 hours of hypoxia. The LA (lactate acid ) content in the supernatant of the culture and the activity of the key enzymes in glycolysis such as hexokinase (HK), phosphofructokinase (PFK) and lactate dehydrogenase (LDH) of both groups of cells were determined at all the time points. RESULTS: (1) After hypoxia, the HK and PFK activities of the rat myocardial cells in H and I groups were obviously increased at the beginning and decreased thereafter when compared with that before hypoxia. While the activities of HK and PFK in H group at 1, 3 and 6 hours after hypoxia were evidently higher than those in I group (P <0.05 or 0.01), and the peak activity of them in H and I groups was 159 +/- 13 U/g vs 133 +/- 55 U/g, and 298 +/- 44 U/g vs 188 +/- 55 U/g, respectively. (2) Compared with normal control (92 +/- 12 U/g), the LDH activity of the cells in H group after hypoxia increased significantly, reaching the peak at 6 hours after hypoxia (2 568 +/- 125 U/g, P < 0. 01), and it decreased thereafter, while that in I group peaked at 3 hours after hypoxia (2125 +/- 126 U/g, P <0.01). The LA content in the culture supernatant in H group increased significantly after hypoxia with the passage of time, while that in I group increased in smaller magnitude (P <0.01). CONCLUSION: High expression of HIF-1alpha in the rat myocardial cells after hypoxia could directly cause continuous enhancement of cell glycolysis, which was beneficial to the protection of myocardial cells under hypoxic condition. PMID- 16383034 TI - [In vitro isolation, cultivation and identification of sebocytes and eccrine sweat gland cells from human fetal skin]. AB - OBJECTIVE: To explore the preliminary methods of in vitro isolation, culture and identification of sebocytes and eccrine sweat gland cells from human fetal skin. METHODS: Human fetal skin was digested with dispase or type II collagenase, and then by micro - sieving to isolate human sebaceous gland and eccrine sweat gland cells. DMEM/F12 (1: 1) was used as the basic culture medium, supplemented with fetal bovine serum, recombinant human epidermal growth factor, L-glutamine, Hydrocortisone, choleratoxin, penicillin and streptomycin as the medium for sebocytes, or fetal bovine serum, recombinant human epidermal growth factor, triiodothyronine, hydrocortisone, insulin, transferrin, sodium selenite to the medium for eccrine sweat gland duct cells. Primary cultures and subcultures were incubated at 37 degrees C in humidified atmosphere of 5% CO2/95% oxygen. Cell morphology was observed by inverted phase contrast microscopy, and the cultured cells were identified with cell clone efficiency determination. The cultured sebocytes were identified with oil red staining and CK4.62, Epithelia Membrane Antigen (EMA) immunohistochemistry staining. The cultured eccrine sweat gland duct cells were identified with CK7, CK19 immunohistochemistry staining. RESULTS: The isolated sebocytes and eccrine sweat gland cells from human fetal skin could grow by adhering to the wall and proliferate in vitro. The cell clone efficiency of human fetal sebocytes was 2.7%, which was obviously lower than that of human fetal keratinocytes (8.0%, P < 0.01). There was no obvious difference in the cell clone efficiency between human fetal eccrine sweat gland cells (7.3%) and human fetal keratinocytes (7.7%, P > 0.05) . The results of oil red staining indicated that a small quantity of lipid droplets in sebocytes, and immunohistochemistry staining of CK4.62, EMA were positive in subculture sebocytes. The immunohistochemistry staining of CK7, CK19 was positive in subculture eccrine sweat gland duct cells. CONCLUSION: In vitro cultured human fetal sebocytes and eccrine sweat gland duct cells displayed the markers and biological characteristics of epithelial lineage, but human fetal sebocytes proliferated more PMID- 16383035 TI - [Influence of recombinant human growth hormone on the prognosis of patients with severe burns a prospective multi-center clinical trial]. AB - OBJECTIVE: To investigate the influence of recombinant human growth hormone (rhGH) on the mortality of the patients with severe burns. METHODS: In a prospective multi-center randomized clinical trial, 207 adult patients with severe burns were enrolled in the study, and they were randomly divided into treatment (T, with subcutaneous injection of rhGH) and placebo control (C, with subcutaneous injection of same amount of isotonic saline) groups. The mortality, incidence of hyperglycemia and sepsis in the two groups were observed. RESULTS: The mortality rate in T group was 0.89% as compared with 5.26% in the C group (P >0.05). Hyperglycemia (blood glucose level over 10 mmol/L) was present in 36.61% of patients in T group but 18.95% in C group (P <0.01). There was no difference in the incidence of sepsis between the two groups (P > 0.05). CONCLUSION: The application of rhGH in appropriate dosage in adult patients with severe burns could be safe, but blood glucose level should be monitored during the administration. PMID- 16383036 TI - [Influence of scald and lipopolysaccharide on gastrointestinal motility]. AB - OBJECTIVE: To investigate the pathogenesis of gastrointestinal motility dysfunction as a result of scald and lipopolysaccharide (LPS) challenge in guinea pigs. METHODS: Thirty guinea pigs were enrolled in the study and were randomly divided into 3 groups:i. e. control (n = 10, with intraperitoneal injection of isotonic saline), scald (n = 10, with 30% TBSA deep partial thickness burn) and LPS (n = 10, with intraperitoneal injection of LPS) groups. Thirty minutes after treatment, all animals were gavaged with carbolic ink. The propelled distance of the ink within the gastrointestinal tract was measured. The intestinal tissue was harvested and homogenized, and the contents of CGRP, Na+-K+-ATP enzyme, Mg2+-ATP enzyme, Ca2+-ATP enzyme, Ca2+-Mg2+-ATP enzyme were determined, and the delta phim of haustra coli smooth muscular cell mitochondria was assessed. RESULTS: The propelled distance of the ink in the gastrointestinal tract in scald (53 +/- 9 cm) and LPS (91 +/- 10 cm) groups was obviously shorter than that in control group (142 +/- 11 cm, P < 0.01). Furthermore, the distance in scald group was shorter than that in LPS group (P < 0.01). The CGRP content in scald and LPS groups [52.0 +/- 39.0 microg/L and 20.0 +/- 23.0 microg/L] was obviously higher than that in control group (0.8 +/-2.0 microg/L, P <0.05 or 0.01), especially in scald group ( P < 0.05). The Na+-K+-ATP enzyme, Mg2+-ATP enzyme, Ca2+-ATP enzyme, Ca2+-Mg2+-ATP enzyme and the delta phim in scald and LPS groups were remarkably lower than those in control group (P <0.005), but there was no difference between scald and LPS groups (P > 0.05). CONCLUSION: The gastrointestinal motility of guinea pigs could obviously be inhibited by scald and LPS, especially by scald. LPS might be the key factor to produce change in the membrane potential of mitochondria of intestinal smooth muscle after severe scald. PMID- 16383037 TI - [The use of diammonium glycyrrhizinate in the treatment of hepatic dysfunction in burn patients]. AB - OBJECTIVE: To observe the protective effect of diammonium glycyrrhizinate on hepatic function in burn patients. METHODS: Twenty burn patients with hepatic dysfunction were enrolled in the study and were randomly divided into 2 groups, i. e. treatment (T, n = 10, with conventional treatment and intravenous infusion of 150 mg diammonium glycyrrhizinate per day for 14 days), and control (C, n = 10, with conventional treatment) groups. The blood samples in both groups were collected before and 7 and 15 days after the treatment. The serum contents of ALT, AST, GGT, ALP and PA in the blood samples were determined and analyzed comparatively. RESULTS: There was obvious difference in the serum contents of ALT, AST, GGT, ALP and PA in the T group before treatment (168 +/- 46 U/L, 104 +/ 29 U/L, 162 +/- 37 U/L, 149 +/- 17 U/L, 310 +/- 35 mg/L, respectively) and 15 days after treatment (51 +/- 9 U/L, 31 +/- 3 U/L, 56 +/- 10 U/L, 103 +/- 9 U/L, 372 +/- 44 mg/L, respectively, P < 0.05). There was no difference in these indices in the C group before and after treatment (P > 0.05). CONCLUSION: Diammonium glycyrrhizinate seemed to be beneficial to the management of postburn hepatic dysfunction with obvious rapid depression of hepatic enzymes. PMID- 16383038 TI - [The relationship between scarless wound healing and the expression of CD68 and CD3 in the immunocytes in fetal skin]. AB - OBJECTIVE: To investigate the difference in the distribution, amount and morphology of immunocytes in fetal skin, normal adult skin and hypertrophic scar, and to probe into their roles in fetal scarless wound healing from the dermatological and immunological point of view. METHODS: Skin specimens obtained from 10 fetuses of induced labor (16 to 33 weeks gestation) due to incipient abortion, 7 adults, and 18 hypertrophic scars in different stages were collected for the detection of the expression and distribution of CD68 (the surface marker of macrophages) and CD3 (the surface marker of T-lymphocytes) with immunohistochemical assay. RESULTS: The cells with positive expression of CD68 (CD68+ macrophages) in fetal skin [(5 +/- 6)/per 400 x visual field] were significantly lower than those in normal adult skin [(23 +/- 4) per/400 x visual field, P <0. 01], and they were obviously lower in normal skin than those in hypertrophic scar [(38 +/- 16)/per 400 x visual field, P < 0.01]. Along with their increase in gestational age, the CD68+ macrophages increased gradually. The cells increased in amount sharply during 24 - 28 gestational weeks, and then the increase slowed down after the 28th gestational week. The lymphocytes with CD3+ expression were not found in all the fetal stages, but were found in small amounts in adult skin [(24 +/- 8)/per 400 x visual field] which were mainly located in the epithelial basal lamina. But there were much more CD3+ lymphocytes [(69 +/- 25)/per 400 x visual field] in the HS, assembling usually in sheet form, and were chiefly distributed in dermal papillary layer around the small vessels in the shape of oversleeve. The cells were much more than those in normal adult skin (P <0.01) in terms of number and pigmentation intensity. CONCLUSION: The low content of CD68+ macrophages in fetal skin might be related to certain extent to the scarless skin wound healing. At the same time, the scarless skin wound healing in fetus could be related to the lack of CD3+ lymphocytes in fetal skin. PMID- 16383039 TI - [The expression of platelet-derived growth factor-A and its receptor in acute radiation-induced skin ulcer in rats]. AB - OBJECTIVE: To investigate the dynamic expression of platelet-derived growth factor-A (PDGF-A) and its receptor alpha (PDGFR-alpha) in different acute radiation-induced skin ulcers, and to explore the underlying mechanism involved in retarded healing of the ulcer. METHODS: The model of acute radiation-induced skin ulcers in rats was replicated with 50 Gy 60Co gamma rays to the skin (radiation group, R, n = 55), rats with full - thickness skin excision wounds as control group (T, n = 55), and 5 normal rats to serve as normal control (NC) group. The expression of PDGF-A and PDGFR-alpha protein and PDGF-A mRNA was respectively assessed by means of histochemistry and in situ RT-PCR. RESULTS: No PDGF-A expression was identified in the rat skin in NC group. The expression of PDGF-A and PDGFR were reduced in R group during inflammatory responsive and granulation formation periods (14 - 28 days after radiation, the IA value of PDGF A varied from 14.0 +/- 1.2 to 20.3 +/- 1.2 compared with that in T group in which the IA value of PDGF-A at the same period (3 - 9 days after injury) varied from 20.0 +/- 1.6 to 28.3 +/- 1.0, and reduced gradually during scar formation period (55 days after radiation). CONCLUSION: The reduction of PDGF-A and PDGFR expression may be partially involved in the mechanism of retarded healing of acute radiation-induced skin ulcers. PMID- 16383040 TI - [Bio-inductive effects of inorganic elements on skin wound healing]. AB - OBJECTIVE: To explore the bio-inductive effects of inorganic elements (Dermlin) on the human epithelial proliferation and differentiation and their promoting effects on skin wound healing. METHODS: 1 ). Cellular test: Normal human skin epithelial cells were cultured with 20 g/L Dermlin supplemented culture medium (E group) and regular culture medium (C group), respectively. The cell proliferation rate and the expressions of type IV collagen and epidermal growth factor (EGF) in the supernatant were determined in 12 and 20 post culture days (PCD). (2). Animal test: Self-consubstantiality control was employed in the study. Sixty Sprague - Dawley rats were inflicted with two symmetric 10% TBSA of superficial or deep partial thickness scald on the back of each rat, and were divided into control[ C, with topical application of silver sulfadiazine (SD - Ag) cream to the wounds] and treatment (T, with 1 g/100 cm2 Dermlin topical application to the wounds) groups. The pathological changes in wound skin were observed and the wound healing rate was calculated on 3, 5, 7, 10, 14 and 18 post treatment day (PTD). (3). Randomized, double-blinded and consubstantiality control method was employed in the clinical trial. Ninety patients were enrolled in the clinical study, among them 30 cases with 60 donor site wounds, 30 with 60 superficial and 30 with 60 deep partial thickness burn wounds were included. Dermlin in dose of 1 g/100 cm2 was applied to the wounds in T group and SD - Ag cream in C group for up to 18 days. Furthermore, sixty patients with diabetic foot ulcers were included for 1 g/100 cm2 Dermlin treatment. The wound healing rate was observed. And the blood and urine test and the indices of hepatic and renal function were determined. RESULTS: 1). Cellular test: The cell proliferation rate and the expression of type IV collagen and EGF in the culture supernatant were obviously higher than those in control group at the same time points (P < 0.01). 2). Animal test: Hyperplastic granulation tissue occurred in the rat wound in the T group since 5 PTD, while that occurred in the C group since 7 PTD. The healing rate of superficial thickness wound in T group on 7, 10, 14 PTD, and that of deep partial thickness wound in T group on 5, 10, 18 PTD were obviously higher than that in the C group (P <0.05). 3). Clinical study indicated that the wound healing rate of the patients with superficial or deep partial thickness scald in the T group was evidently higher than that in the C group on 5 and 10 PTD (P <0.05), but the wound healing time of the superficial, deep partial thickness wound and donor site wound in the T group was significantly shorter than that in the C group (P < 0.05). Before treatment, the square of the ulcers on the foot of the patients with diabetic was (39 +/- 28) cm2, and it was reduced to (19 +/- 23) cm2 2 weeks later, with the therapeutic efficacy reaching 62.5% . For all patients, no obvious change was found in the blood test and hepatic and renal function indices. CONCLUSION: The inorganic element (Dermlin) is beneficial to wound healing and to the proliferation and differentiation of epithelial cells. PMID- 16383041 TI - [Purification of human endothelial overexpressed lipopolysaccharide-associated factor 1 protein]. AB - OBJECTIVE: To investigate the feasibility of obtaining of a highly pure protein of human endothelial overexpressed lipopolysaccharide-associated factor 1 (EOLA1) with metal chelation chromatography. METHODS: Inclusion bodies of the E. coli transformed with EOLA1 gene were extracted and washed with BugBuster Protein Extraction Reagent. The primary purified products were purified by His. Bind Resin Chromatography under denaturing condition and dialyzed for renaturation, and then were analyzed with SDS-PAGE, Western blotting and peptide mass fingerprinting (PMF). RESULTS: EOLA1 was mainly expressed in E. coli as insoluble inclusion bodies. The protein content in the primary extracted inclusion bodies accounted for over 75%, and it accounted for more than 90% after chromatography and renaturation. It was indicated by PMF that the targeted protein peptide overlaid many of designed protein peptide. CONCLUSION: The method of EOLA1 protein purification and renaturation was convenient and efficient, and by this method sufficient amount of highly pure EOLA1 protein could be obtained for the preparation of EOLA1 monoclonal antibody and for the study of its gene function. PMID- 16383042 TI - [Influence of different thawing temperature on the morphology and type I collagen metabolism of the human fibroblasts processed at - 10 degrees C in vitro]. AB - OBJECTIVE: To explore the influence of different thawing temperatures on the morphology and type I collagen metabolism of the human fibroblasts processed at - 10 degrees C in vitro. METHODS: In vitro cultured human fibroblasts were randomly divided into control, 20 degrees C thawing, and 37 degrees C thawing groups. After being frozen at -10 degrees C, the cells in the latter two groups were thawed at 20 degrees C and 37 degrees C, respectively. The cell proliferation was assessed with MTT method and was expressed by absorption under 570nm (A ), The morphological change of the cells was observed with inverted phase contrast microscope, the change in the intracellular content of collagen was determined with immunohistochemistry, and the extracellular content of collagen was assayed with ELISA. RESULTS: In 20 degrees C thawing group, the absorbance decreased at first and increased thereafter, and they were obviously lower than that before freezing (0.95 +/- 0.16, P < 0.05 or 0.01). Cell dehydration and shrinking, cytoplasm loss and increased ratio of cytoplasm to nucleus were found in the survived fibroblasts. The cells proliferated actively at 72 and 96 hours after injury, with increased mitotic index and disordered arrangement. Compared with that before freezing (96.4 +/- 2.9) , the extracellular collagen content increased at first, decreased thereafter, and increased again slowly later (P < 0.05), while the intracellular collagen content decreased at first and increased thereafter (P < 0.05). The collagen metabolism in 37 degrees C thawing group was no difference compared with that in control group. Some cells undergone a floating period before adhering to the culture dish walls. CONCLUSION: Cell dehydration after low temperature treatment could protect the cells from damage. Proper thawing temperature could be beneficial to the cell resuscitation. Comparing with slow thawing, rapid thawing could minimize the cell damage. PMID- 16383043 TI - [Influence of fluorescent protein expression on the proliferation of NIH3T3 cells in vitro]. AB - OBJECTIVE: To investigate the influence of fluorescent protein expression on the proliferation of murine NIH3T3 cells, so as to provide a theoretical basis for cell tracing technology. METHODS: NIH3T3 cells were cultured in vitro, and were randomly divided into control, pLEGFP-N1 (with transfection of pLEGFP-N1 retroviral vector), pEGFP-N1 (with transfection of pEGFP-N1 vector) and pDsRed2 C1 (with transfection of pDsRed2-C1 vector) groups. Then the cells were screened by G418 for 3 weeks. The changes in cell adhesive rate were observed and the population doubling times was determined by growth curve. RESULTS: There was obvious fluorescent protein expression in the transfected NIH3T3 cells after G418 selection, and the highest percentage of labeled NIH3T3 cells was found in pLEGFP N1 group. The population doubling time in pDsRed2-C1 (40.3+/-0.7 h) , PEGFP-N1 (39.6 +/- 0.6 h) and pLEGFP-N1 (36.5 +/- 0.7 h) groups was evidently longer than that in control (27.9 +/- 0.6 h, P < 0.01), with high adhesive rate in each group. CONCLUSION: The expression of fluorescent protein exhibited some inhibitory effect on the proliferation of NIH3T3 cells in vitro. Since the inhibitory effect by retroviral vector was weaker compared with eukaryotic vector, it should be the first choice for fluorescent protein labeling during cell transplantation. PMID- 16383044 TI - Treating asymptomatic patients. PMID- 16383045 TI - Footing the bill. PMID- 16383046 TI - Disparities and diversity. PMID- 16383047 TI - Managing the care of patients with bisphosphonate-associated osteonecrosis: an American Academy of Oral Medicine position paper. AB - BACKGROUND: This position paper addresses the prevention of bisphosphonate associated osteonecrosis (BON) and the management of care of patients with cancer and/or osteoporosis who are receiving bisphosphonates and who have BON or are at risk of developing it. METHODS: The authors reviewed the literature available on this newly described oral complication. Information of interest included bisphosphonates, the medications associated with this oral complication; the patient population at risk of developing BON and the diseases being treated with this class of medications; the clinical presentation of the oral lesions; guidelines for managing the care of patients who develop BON; the prevention of this complication based on current knowledge; and recommendations for routine dental treatment of patients receiving bisphosphonates. RESULTS: There is strong evidence that bisphosphonate therapy is the common link in patients with BON. The pathobiological mechanism leading to BON may have to do with the inhibition of bone remodeling and decreased intraosseous blood flow caused by bisphosphonates. People at risk include patients with multiple myeloma and patients with cancer metastatic to bone who are receiving intravenous bisphosphonates, as well as patients taking bisphosphonates for osteoporosis. The risk of developing complications appears to increase with time of use of the medication. There are no guidelines based on evidence, and the clinical management of the oral complication is based on expert opinion. CONCLUSION: Prevention of BON is the best approach to management of this complication. Existing protocols to manage the care of patients who will receive radiation therapy or chemotherapy may be used until specific guidelines for BON are developed. PMID- 16383048 TI - Bisphosphonate-associated osteonecrosis of the jaws: a review of current knowledge. AB - BACKGROUND: Increasing numbers of dental patients are taking bisphosphonate medications for a variety of indications. These drugs may be associated with poor healing, spontaneous intraoral ulceration and bone necrosis in the oral and maxillofacial region. CASE DESCRIPTION: The authors describe a case of osteonecroSis of the jaws in a patient receiving long-term bisphosphonate therapy for cancer. They offer recommendations for management and prevention of oral complications. CONCLUSION AND CLINICAL IMPLICATIONS: It is important that clinicians are aware of the association between bisphosphonate treatment and delayed wound healing and osteonecrosis of the jaws. They should consider referring patients in this population to specialists for even the most routine oral surgery. Clinicians should perform a thorough oral examination in patients before they begin any chemotherapy regimen. PMID- 16383049 TI - Osteonecrosis of the jaws in patients with a history of receiving bisphosphonate therapy: strategies for prevention and early recognition. AB - BACKGROUND: Bisphosphonates, inhibitors of osteoclasts, have been shown to alleviate many of the devastating consequences associated with metastatic bone disease. However, recent reports have shown that bisphosphonates may cause osteonecrosis of the jaws. Since the publication of these initial reports, the authors have treated several patients with osteonecrosis of the jaws who had a history of receiving bisphosphonate therapy. METHODS: The authors reviewed the medical records of patients who visited their clinic between September 2003 and December 2004 and who had osteonecrosis of the jaws and a history of having received bisphosphonate therapy but no irradiation to the head and neck. RESULTS: Eleven patients (four female and seven male) with a mean age of 69 years were included in this report. They had received bisphosphonate therapy for a mean duration of 34 months. Radiographic data showed loss of bone density at sites of osteonecrosis, and histologic examination demonstrated necrosis of bone without evidence of metastases. CONCLUSIONS AND CLINICAL IMPLICATIONS: Further research is required for better understanding of the role of bisphosphonates in the development of osteonecrosis of the jaws. Until more is known, the authors recommend that measures be taken to prevent osteonecrosis those at risk, including identifying patients with a history of having received bisphosphonate therapy before they undergo dental surgery. To help identify such patients, the authors propose the use of a screening questionnaire. When feasible, physicians should consult with their patients' general dentists or oral surgeons before patients begin bisphosphonate therapy. PMID- 16383050 TI - Digital imaging fiber-optic trans-illumination, F-speed radiographic film and depth of approximal lesions. AB - BACKGROUND: Detection of early carious lesions is a prerequisite to an optimal preventive and minimal surgical intervention strategy. The authors conducted this study to determine whether Digital Imaging Fiber-Optic Trans-Illumination (DIFOTI) (Electro-Optical Sciences, Irvington, N.Y.) could be useful in evaluating early approximal lesions and to compare radiographs produced with F speed film with both histologic lesion depth and cavitation. METHODS: The authors created artificial approximal lesions in vitro in extracted teeth over 14 weeks and imaged them using a "bitewing-like" view every two weeks with DIFOTI and F speed radiographic film. At the end of the 14 weeks, the authors examined the lesions for surface cavitation using visual and tactile methods. They then thin sectioned the lesions and subjected them to histologic analysis using polarized light microscopy (PLM). RESULTS: DIFOTI was not able to measure the depth of a lesion in any of the samples. It was, however, able to show surface changes associated with early demineralization as early as two weeks. The depth of a lesion measured using F-speed radiographic film was not statistically different from the depth of a lesion measured with PLM histologic analysis (P > .05). None of the lesions showed any signs of surface cavitation after 14 weeks of demineralization. CONCLUSIONS AND CLINICAL IMPLICATIONS: DIFOTI technology should not be used to decide between surgical or chemical treatment strategies based on lesion depth. We found that F-speed radiographic film was accurate in the approximating the depth of the lesion histologically. Ideally, the clinical decision whether to cut the tooth should be made based on cavitation rather than histologic lesion depth. PMID- 16383051 TI - Removal of oral biofilms by bubbles: the effect of bubble impingement angle and sonic waves. AB - BACKGROUND: Previous research showed that the collision of bubbles with biofilm could remove biofilm from a surface. However, the effectiveness of biofilm removal by bubbles and sonic waves had not been determined. METHODS: The authors mounted Streptococcus mutans biofilms in a chamber containing artificial saliva and exposed them to bubbles and sonic waves. They generated sonic waves via an oscillator at the frequencies and acoustic intensities of sonic toothbrushes. They also mounted biofilms at different angles to measure the effect of a bubble's impingement angle. RESULTS: The presence of sound had no significant effect on the amount of biofilm removed (F = 0.51). There was no statistically significant difference in the amount of biofilm removed with respect to the angle at which bubbles impinged on the biofilm (F = 0.65). The authors performed analysis of variance tests to determine whether the difference in the amount of biofilm removal was significant (F < 0.05) in the presence or absence of sound and at the different angles tested. CONCLUSIONS: The collision of a bubble at any angle between 5 and 45 degrees was equally effective in removing biofilm. The addition of sound to a bubble stream at the tested intensities had a negligible effect. CLINICAL IMPLICATIONS: Bubbles produced by dental instruments are effective in removing biofilm from the surface of the teeth, regardless of the angle at which the bubble impinges on the biofilm. PMID- 16383052 TI - Noncarious cervical lesions in adults: prevalence and occlusal aspects. AB - BACKGROUND: The factors that induce the formation of noncarious lesions are not understood fully, particularly those that are related to occlusal aspects. The authors conducted a study to evaluate the prevalence of noncarious cervical lesions in adults and their association with occlusal aspects. METHODS: The authors examined 70 people (35 men and 35 women) aged 25 to 45 years to determine the presence and type of noncarious cervical lesions, wear facets, tooth contacts in maximal intercuspal position, and lateral and protrusive movements. The assessment involved a questionnaire and clinical examination. RESULTS: Among the teeth the authors evaluated, 17.23 percent had cervical lesions, 80.28 percent of which had wear facets (P < .01). The authors found a significant difference between the prevalence of noncarious lesions and the presence of wear facets (P = .0484). CONCLUSIONS: The authors found that cervical lesions were related significantly to wear facets. These findings strengthen evidence for the role of occlusal forces as an etiologic factor for noncarious lesions. CLINICAL IMPLICATIONS: The presence of wear facets should be considered in the treatment of noncarious cervical lesions. PMID- 16383053 TI - The integration of clinical research into dental therapeutics: making treatment decisions. AB - BACKGROUND: Clinicians make informed therapeutic decisions in part by assimilating evidence that forms the basis for generally accepted clinical knowledge. Similar to professional training for the practice of dentistry, this requires knowledge of the quality of the evidence and its translation to clinical practice. DESCRIPTION: Evidence-based treatment decisions rely on the ability to understand the differences between clinical practice and clinical research, to critically evaluate the quality of evidence that supports a generally accepted clinical practice, understanding what contributes to the perception of clinical success, and how to select the most appropriate therapy for use in clinical practice. CLINICAL IMPLICATIONS: Therapeutic decision making is highly dependent on the quality of evidence that is considered when making a judgment and applying it to patient care. Criteria for evaluating clinical research and the findings evolve over time, necessitating periodic review of the foundations on which therapeutic decisions are based. PMID- 16383054 TI - How to kill a tooth. AB - Many dentists report increased activity in endodontic therapy. Although retention of teeth through most of life by many patients and the aging population are potential reasons for this increase, it is possible that some of the new dental procedures, instruments or materials that have become popular in recent years also are significant causative factors. Several potential preventable reasons for increased pulpal death and the resultant need for endodontic therapy are discussed in this article. PMID- 16383055 TI - The clinical performance of CAD/CAM-generated composite inlays. AB - BACKGROUND: The authors conducted a study to evaluate the longitudinal clinical performance of a resin-based composite (Paradigm, 3M ESPE, St. Paul, Minn.) for computer-aided design/computer-aided manufacturing (CAD/CAM)-generated adhesive inlays. METHODS: The researchers used a CAD/CAM unit (CEREC 2, Sirona Dental Systems, Bensheim Germany) to fabricate 40 porcelain (Vita Mark II, Vita Zahnfabrik, Bad Sackingen, Germany) and 40 resin-based composite (Paradigm, 3M ESPE) inlays. Both restorative materials were cemented with a total-etch technique using Single Bond dental adhesive (3M ESPE) and a dual-cured resin cement (RelyX ARC Adhesive Resin Cement, 3M ESPE). Two examiners evaluated the inlays using modified U.S. Public Health Service (PHS) criteria at six months, one year, two years and three years. RESULTS: No sensitivity was reported for either material at any recall period. There was no significant difference between the two materials relative to margin adaptation at three years. Margin adaptation initially was very good for both materials, with an increase in margin detection due to apparent wear of the resin-based composite luting agent. There was a significant difference in color match between the two materials at three years, with 91.4 percent of the resin-based composite inlays and 58.8 percent of the porcelain inlays rated Alfa. CONCLUSIONS AND CLINICAL IMPLICATIONS: The resin based composite inlays had a significantly better color match at three years than did the porcelain inlays. Resin-based composite CAD/CAM inlays performed as well as porcelain CAD/CAM inlays after three years of clinical service. PMID- 16383056 TI - The treatment effects of Invisalign orthodontic aligners: a systematic review. AB - BACKGROUND: The authors conducted a systematic review of the literature to determine the treatment effects of the Invisalign orthodontic system (Align Technology), Santa Clara, Calif.). TYPES OF STUDIES REVIEWED: The authors reviewed clinical trials that assessed Invisalign's treatment effects in nongrowing patients. They did not consider trials involving surgical or other simultaneous fixed or removable orthodontic treatment interventions. RESULTS: The authors searched electronic databases (PubMed, MEDLINE, MEDLINE In-Process & Other Non-Indexed Citations, Evidence Based Medicine Reviews, EMBASE Excerpta Medica, Thomsen's ISI Web of Science and LILACS) with the help of a senior health sciences librarian. They used "Invisalign" as the sole search term, and 22 documents appeared in the combined search. Thereafter, they used "clinical trials," "humans" and "Invisalign treatment effects" as abstract selection criteria. Only two published articles met these inclusion criteria, though after reading the actual articles, the authors determined that they did not adequately evaluate Invisalign treatment effects. Both articles identified methodological issues. CLINICAL IMPLICATIONS: The inadequately designed studies the authors found represented only a lower level of evidence (level II). Therefore, the authors found that no strong conclusions could be made regarding the treatment effects of Invisalign appliances. Future prospective randomized clinical trials are required to support, with sound scientific evidence, the claims about Invisalign's treatment effects. Clinicians will have to rely on their Invisalign clinical experience, the opinions of experts and the limited published evidence when using Invisalign appliances. PMID- 16383057 TI - Continuing education: balancing team development in four directions. AB - Continual growth over time will lead to the rewarding practice every dentist envisions. A balanced CE plan that includes programming and dedicated funding for the development of interpersonal, clinical and financial skills leads to greater practice success and professional fulfillment. A substantial CE investment for the dentist and staff members has been the choice of many of the most successful dental practices. PMID- 16383058 TI - Peer review under the Health Care Quality Improvement Act. PMID- 16383059 TI - Ethical moment: how can I handle a rude patient? PMID- 16383060 TI - For the dental patient. Fluoride: nature's cavity fighter. PMID- 16383061 TI - Impressions of people with gender-ambiguous male or female first names. AB - Undergraduates (12 men, 12 women) read a scenario in which they formed an impression of nine people who had left their first name on an answering machine. Participants rated the extent to which seven characteristics (Ethical, Caring, Popular, Cheerful, Successful, Masculine, Feminine) applied to people whose first names were gender-ambiguous (e.g., Chris), male (e.g., Ken) or female (e.g., Pam). People with gender-ambiguous names were rated less Ethical than those with female names, and people with gender-ambiguous names and male names were rated less Caring, less Cheerful, and less Feminine than those with female names. These results are consistent with the idea that there is a bias towards assuming that a person of unspecified sex is a male. PMID- 16383062 TI - Evaluating current deficit theories of poor reading: role of phonological processing, naming speed, balance automaticity, rapid verbal perception and working memory. AB - To clarify the nature of cognitive deficits experienced by poor readers, 9-10-yr. old poor readers were matched against 9 chronological age and 9 younger reading age-matched controls screened and selected from regular classrooms. Poor readers performed significantly more poorly than chronological age-matched peers on digit naming speed, spoonerisms, and nonsense word reading. Poor readers were also significantly poorer than reading age-matched controls on nonword reading but were significantly better than reading age-matched controls on postural stability. Analyses of effect sizes were consistent with these findings, showing strong effects for digit naming speed, spoonerisms, and nonword reading. However, effect size analysis also suggested that poor readers experienced moderate difficulties with balance automatisation but did not show verbal speech perception deficits relative to either control PMID- 16383063 TI - Estimating curvature of nondifferentiable functions and complex shape contours. AB - Salvadori and Luccio demonstrated that, for the purpose of psychophysical analysis, the curvature of a point on a continuous single-valued function can be quantified as the inverse ray of the circle osculating the function at the point of interest. While this method is mathematically sound, it does not allow for estimation of curvature in complex shapes containing discontinuities and irregularities such as sharp corners, closed contours, or line intersections- features that commonly occur in normal visual experience. In this paper, a simple modification of this algorithm is presented which overcomes these limitations by using discrete rather than continuous contour sampling, thereby allowing estimation of curvature for a wider variety of shape contours. PMID- 16383064 TI - An analysis of players' consistency among professional golfers: a longitudinal study. AB - This study followed the performance of individual professional golfers who played from 1997 to 2000 on the Professional Golfers Association Tour (PGA Tour, N=35), Ladies Professional Golfers Association Tour (LPGA Tour, N = 25), and Senior Professional Golfers Association Tour (SPGA Tour, N = 31) and investigated individual players' consistency in round-to-round performance within tournaments. Although the number of players showing consistency for most years exceeded chance expectations, evidence for round-to-round consistency from one year to the next for any player was small. Such evidence suggests that variability in player performance is more the rule than the exception for players on the three American professional tours. PMID- 16383065 TI - Relationship between magical ideation and noctcaelador. AB - This study explored the relationship between magical ideation and "noctcaelador" (strong interest in, and psychological attachment to, the night sky). 210 university students completed Eckblad and Chapman's 1983 Magical Ideation Scale and Kelly's 2004 Noctcaelador Inventory. Scores on the two scales were significantly positively related and accounted for 14% of the common variance. Based on this operational definition of magical ideation, a strong interest in the night-sky might be associated with uncommon beliefs and reports of unusual perceptual experience. Researchers must clarify and define these concepts to study possible relations. PMID- 16383066 TI - Changes in recovery-stress state and performance in elite rowers during preparation for major competitions. AB - This study investigated changes in the perceived recovery-stress state and performance in 6 highly trained Estonian rowers preparing for major competitions and representing Estonia in double sculls (age: M = 23.0, SD = 1.4 yr.; height: M = 198.0, SD = 0.0 cm; body mass: M = 97.1, SD = 6.4 kg), and quadruple sculls (age: M = 21.5, SD = 2.5 yr.; height: M = 189.5, SD = 5.4 cm; body mass: M = 90.9, SD = 4.8 kg). The subjects were training regularly and had been doing so for the last 10.5 yr. (double scullers, SD = 2.1 yr.) and 7.5 yr. (quadruple scullers, SD = 3.1 yr.), respectively. The Recovery-Stress Questionnaire for Athletes of Kellmann and Kallus was given two days before World Cup competitions. Standardized Stress and Standardized Recovery scores and RESTQ-Index were computed. In addition, resting cortisol in blood was measured at the same time. The performance of both boat classes constantly decreased from World Cup 1 to World Cup 3 as indicated by percent losses for winning boats. Resting cortisol and RESTQ-Index increased and decreased, respectively, at the same time. For double scullers, a decrease in RESTQ-Index mainly reflected the large decrease in the Standardized Recovery score. A decrease in RESTQ-Index was mainly based on a large increase in the Standardized Stress score for rowers performing in a quadruple scull. Present results indicate that the recovery-stress state affects performance of rowers in international competitions. RESTQ-Index could be used as the athletes' resource measure, while the Standardized Stress and Standardized Recovery scores reflect the athletes' extent of stress and recovery separately. PMID- 16383067 TI - The "birthday blues" in a sample of major league baseball players' suicides. AB - Among 74 major league baseball players known to have committed suicide, 17 occurred within 28 days of the players' birthdays as compared to an expected number of 11.4. PMID- 16383068 TI - Development of a portable psychometric testing device for use in the field: an alcohol investigation. AB - Cognitive and psychomotor performance have traditionally been assessed in the laboratory. There is a need for an objective portable assessment tool to assess cognitive and psychomotor performance. This study investigated the viability of a portable psychometric test battery, in a controlled laboratory environment, possibly leading to use in the field. A randomised, double-blind placebo controlled, three-way crossover design was employed. 16 subjects received 50 mg/100 ml and 80 mg/100 ml of alcohol and alcohol placebo. Performance was assessed with a tracking task, and an attention task presented on a small ruggedised handheld computer. The attention task showed no significant training effects; however, an element of the tracking task did. Statistical significance, effect size, and test-retest reliability analyses are presented indicating sensitivity of the portable psychometric test battery to the impairing effects of two separate doses of alcohol. Ability to undertake wide-scale impairment testing in the field with meaningful results in the absence of baseline data collection may have wide reaching implications, particularly in relation to the assessment of drivers impaired by drug use. PMID- 16383069 TI - Sex differences in Portuguese lonely hearts advertisements. AB - Advertisements from "Lonely Hearts" columns in the major daily Portuguese newspaper (Jornal de Noticias) were used to test hypotheses about the mate preferences of men and women. A total of 484 advertisements were coded for demographic descriptors and offers of and appeals for attractiveness, financial security, sincerity, expressiveness, and instrumentality, e.g., intelligence and ambition. Some results supported social exchange and evolutionary predictions: men sought younger women and offered security; women sought older men with status and resources. However, other results challenged such predictions: attractiveness and expressiveness did not differ by sex. PMID- 16383070 TI - Effect of stimulant medication on lower extremity response time of boys with attention deficit hyperactivity disorder. AB - Children with attention deficit hyperactivity disorder (ADHD) have been labeled as "inefficient movers"; however, little research has examined the effect of stimulant medication on lower extremity movements. 16 boys, 11 to 13 years old, with ADHD performed a lower-limb choice-response time task, both on and off medication. When nonmedicated, children had significantly slower reaction times to all three targets and significantly slower movement times for the contralateral and midline movements. For both conditions, children had significantly faster movement time when using the right leg than the left leg. These findings suggest that movement characteristics of children with ADHD are different under medicated and nonmedicated situations. PMID- 16383071 TI - The domestic behavior of northerners and southerners. AB - The northern and southern regions of a sample of 26 nations did not differ in birth, marriage, or divorce rates. PMID- 16383072 TI - Reliability and factorial validity of the standard progressive matrices among Kuwaiti children ages 8 to 15 years. AB - The Raven Standard Progressive Matrices was administered to a sample of 6,529 children in Kuwait ranging in age from 8 to 15 years. Test-retest reliability (N = 968) ranged between .69 and .85, while Cronbach coefficients alpha ranged from .88 to .93, showing from acceptable to good temporal stability and from good to high internal consistency. The loadings of the five sets of matrices on the only salient factor ranged from .73 to .89 indicating the good factorial validity of the scale. The rtest seems useful in the Kuwaiti context. PMID- 16383073 TI - Effects of modeling and topic stimulus on self-referent touching. AB - Self-touching is defined as self-directed touches, e.g., scratching or rubbing, that have no apparent conscious motivation. Earlier research on self-touching focused on internal processes associated with self-touching, e.g., mental conflict, while more recent research has considered external motivators, such as an auditory or visual presentation of a provocative stimulus, e.g., leeches. Here, 110 undergraduate psychology students (68 women, 42 men; M age = 18.7 yr., SD = 1.0) were shown a video about chiggers to replicate previous findings, while simultaneously studying the social influence of self-touching modeled by the presenter. A significant interaction was found as modeled self-touching significantly increased participants' self-touching when they were also attending to the video about chiggers. The results are discussed in terms of potential processes underlying this behavior and directions for research. PMID- 16383074 TI - Cross-cultural differences for three visual memory tasks in Brazilian children. AB - Norms for three visual memory tasks, including Corsi's block tapping test and the BEM 144 complex figures and visual recognition, were developed for neuropsychological assessment in Brazilian children. The tasks were measured in 127 children ages 7 to 10 years from rural and urban areas of the States of Sao Paulo and Minas Gerais. Analysis indicated age-related but not sex-related differences. A cross-cultural effect was observed in relation to copying and recall of Complex pictures. Different performances between rural and urban children were noted. PMID- 16383075 TI - National character and sexual behavior. AB - Citizens of 22 nations which were rated as more extraverted had more sexual partners and had their first sexual encounter at a younger age than those in nations who were rated as less extraverted. PMID- 16383076 TI - Gesture production and text structure. AB - In the present pilot study, we examined the relationship between gesture and verbal production. Subjects were asked to read a story and then to narrate what they remembered. We analyzed verbal production according to Kintsch's procedure and obtained a list of hierarchically organized propositions that represented the meaning of subject's narration. Propositions were subdivided into High- and Low level propositions. Further, we analyzed gesture production by identifying hand or arm movement that accompanied subject's narration. Each gesture was associated with the word simultaneously produced, and then with the proposition to which the word belonged. Analysis showed subjects were more likely to produce gestures in association with High- rather than with Low-level propositions. High-level propositions may elicit the production of gestures which, in turn, mark the importance of verbal meaning conveyed simultaneously. PMID- 16383077 TI - Sex differences on use of associative and dissociative cognitive strategies among male and female athletes. AB - This study assessed whether men and women differed in using associative and dissociative cognitive strategies during athletic performance. Athletes (31 men, M age = 23.2 yr., SD = 3.9 and 29 women, M age = 22.9 yr., SD = 4.3) who practiced endurance activities (running, swimming, and cycling) were considered high-level performers because they participated in national or international competition. The athletes were interviewed, and Schomer's 1986 method of measurement was used to evaluate and quantify two cognitive strategies. Most specifically, categories of association concerned the way the athlete paid close attention to bodily signals, and categories of dissociation described how the athlete shunned sensory inputs. Analysis of variance and the t test showed that women tend to be more dissociative than men and men more associative than women. The results suggest that pain perception in these sports may be a function of sex. PMID- 16383078 TI - DRM paradigm for people's name lures and effects of delaying conscious thought of the lure at encoding. AB - The experiment was conducted to test whether or not presentation order is an important factor to explain why Critical Lures of people's names are falsely recalled relatively rarely in the Deese-Roediger-McDermott paradigm. For half of the participants, the most highly associated items were presented at the start of the list (Standard condition). For the other half, presentation of the most strongly associated items was delayed until the middle of the list (Delayed condition). This manipulation had no significant effect on the probability of falsely recalled Critical Lures or on that of veridical recall of list items, but in the Delayed condition the mean number of intrusions other than the Critical Lures was higher than that in the Standard condition. PMID- 16383079 TI - Updated internal consistencies and intercorrelations for the inventory of learning processes. AB - Present data on the internal consistencies and interrelationships of the subscales of the Inventory of Learning Processes nearly 30 years after its development show these to be similar to the original values. Researchers can continue to use the original version of the inventory in studies investigating learning styles. PMID- 16383080 TI - Effects of sleep deprivation on auditory and visual memory tasks. AB - Probe recognition tasks have shown the effects of sleep deprivation following a full night of sleep loss. The current study investigated shorter durations of deprivation by testing 11 subjects for accuracy and response time every 2 hr. from 10 p.m. through 8 a.m. We replicated Elkin and Murray's auditory single probe recognition task using the number triplets and added two visual tasks with number and shape triplets. Series of six stimuli were each followed by a probe, which was presented after 2.5 sec. as a short delay or 20 sec. as a long delay. Accuracy performance showed a significant decrease for the long delay beginning after 4 a.m. for the two visual tasks. Response times were significantly slower for the visual shapes task using the short delay. Visual tasks, especially shapes, may be more prone to disruption by sleep deprivation, given the visual information load and the briefness of iconic memory. PMID- 16383081 TI - A FORTRAN program for computing the exact variance of weighted kappa. AB - An algorithm and associated FORTRAN program are provided for the exact variance of weighted kappa. Program VARKAP provides the weighted kappa test statistic, the exact variance of weighted kappa, a Z score, one-sided lower- and upper-tail N(0,1) probability values, and the two-tail N(0,1) probability value. PMID- 16383082 TI - Effect of color schemes and environmental sensitivity on job satisfaction and perceived performance. AB - Effects of interior office color and individual stimulus screening ability, i.e. instinctive perceptual filtering of irrelevant stimuli, on perceived performance and job satisfaction were examined on various outcome measures over a 4-day work week in a laboratory setting. Workers performed specific tasks and worked regular hours for 4 consecutive days in the same office. They were separated into three groups according to their ability to ignore irrelevant stimuli in the interior space. Those in the white and predominantly blue-green offices reported higher perceived job performance and satisfaction than those in the predominantly red office regardless of stimulus screening ability. Workers with high to moderate stimulus screening ability indicated greater perceived performance and job satisfaction than did workers with low stimulus screening ability. Results for color schemes are discussed in terms of preference and social expectations. PMID- 16383083 TI - Controlled exertion of force by developmentally delayed young men and women. AB - Present purposes were to examine the characteristics of controlled force exertion in 28 developmentally delayed young people (14 men, 14 women), and sex differences compared to 28 normal young students (14 men, 14 women). The subjects matched their submaximal grip strength to changing demand values displayed in a bar chart on the display of a personal computer. The total sum of the differences between the demand value and grip exertion value for 25 sec. was used as an evaluation parameter for the test. The controlled force exertion was significantly poorer for he developmentally delayed group than for controls, and there were large individual differences. The developmentally delayed men scored poorer than women in coordination. Like the controls, the means between trials did not decrease significantly. For these developmentally delayed subjects, performance did not improve after only a few trials. The controlled force exertion test is useful as a voluntary movement-function test for developmentally delayed subjects. PMID- 16383084 TI - Absolutism in the diary of a suicide. AB - The incidence of three words indicative of irrational thinking (perfect, always, and never) did not increase in the diary of a suicide over the course of the year preceding her suicide. PMID- 16383085 TI - Relationship of personalized jerseys and aggression in women's ice hockey. AB - The present study examined the relationship between aggression and players' names on uniforms in collegiate women's ice hockey. Aggression was defined as mean penalty minutes per game. Information, i.e., win/loss record, penalties, and names on uniforms, about the 2002-2003 season women's ice hockey team was obtained via e-mail from 53 of 72 (74% return rate) sports information directors (Division I = 23, Division II = 2, Division III = 28). Analysis indicated that teams with personalized jerseys had significantly more penalty minutes per game than teams without personalized jerseys. However, as the majority of the teams with personalized jerseys were Division I teams and the majority of the teams without personalized jerseys were Division III teams, it is unclear whether results were due to personalized jersey or competition level of play. PMID- 16383086 TI - Pilot study of telephone tutoring in reading skills. AB - This study tested the feasibility of tutoring children in reading via telephone sessions. 19 children received tutoring from any of 6 tutors for an average of 7.6 hr. per month. Initially, these children were an average of 10.4 yr. old, in average Grade 4.8, and averaged 2.9 grade levels behind their grade expectations on the Slosson Oral Reading Test. Before tutoring the children had progressed on the Slosson at an average of 0.44 grade levels per year (95% CI = 0.30 to 0.57). The children participated in telephone tutoring for an average of 1.5 yr. During tutoring the students progressed at an average of 2.0 grade levels per year (95% CI = 1.2 to 2.8). The rate of progress during tutoring was significantly greater than that before tutoring (p<.001). The relations between tutors and students appeared to be positive and pleasant. Telephone tutoring seems to be a practical and feasible service delivery method which should be tested further. PMID- 16383087 TI - One-sided nonparametric tests for ordinal data. AB - Baumgartner, Weiss, and Schindler (1998) introduced a novel non-parametric test for the two-sample comparison that is superior to commonly used tests such as the Wilcoxon rank-sum test. A modification of the novel test statistic can be used for one-sided comparisons based on ordinal data. Such comparisons frequently occur in psychological research, and the Wilcoxon test is often recommended for their analysis. Here, the two tests were compared in a simulation study. According to this study the tests have a similar type I error rate, but the modified Baumgartner-Weiss-Schindler test is more powerful than the Wilcoxon test. PMID- 16383088 TI - Automatic prompting to reduce persistent tongue protrusion in a woman with severe to profound mental retardation. AB - This study assessed whether an intervention approach relying on auditory prompting delivered automatically through a portable device was effective to reduce tongue protrusion in a woman with severe to profound mental retardation. The device involved (a) an optic sensor, i.e., a miniphotocell kept under the lower lip with medical tape, (b) a small signal transmission box, and (c) a Walkman for presenting the prompts. Initially, the automatic prompting condition was combined with occasional praise from a research assistant for having the tongue in the mouth. Analysis showed that the occurrence of tongue protrusion dropped from about 65% of the observation time during the initial baseline to less than 5% through the intervention. The study lasted 4.5 mo. PMID- 16383089 TI - Anxiety and burnout in female collegiate ice hockey and soccer athletes. AB - The purpose of this study was to assess whether athletes who perceived their scores on trait anxiety to be debilitative to performance also experienced higher burnout scores. 84 NCAA Division I female ice hockey and soccer athletes, ages 18 to 23 years, volunteered. Participants were given a trait version of the Competitive State Anxiety Inventory-2 with an added direction scale to collect general perceptions of anxiety. They also completed the Burnout Inventory for Athletes. A one-way multivariate analysis of variance for group anxiety yielded an overall significant effect, with the Facilitative group (n = 40) reporting a lower mean burnout score. Mean intensity of cognitive anxiety was significantly higher for the Debilitative group (n = 44), while mean self-confidence intensity was significantly higher for the Facilitative group. PMID- 16383090 TI - Effects of cooperative learning strategy on undergraduate kinesiology students' learning styles. AB - A growing body of research supports cooperative learning as an effective teaching strategy. A specific cooperative learning strategy, Team-based Learning, was applied to a convenience sample of four undergraduate sophomore-level motor behavior courses over four semesters from Fall 2002 to Spring 2004 to examine whether this strategy would affect students' learning styles. The data from the Grasha-Reichmann Student Learning Style Scales indicated that this teaching strategy was associated with a significant decrease in the negative Avoidant and Dependent learning styles and an improvement in the positive Participant learning style. PMID- 16383091 TI - Variable practice in learning the forehand drive in tennis. AB - One of the most important factors that has occupied researchers in motor learning is the method of practicing motor skills. This study examined the effect of variable practice in learning the forehand drive in tennis by children and adolescents. Variability of practice was achieved by the alternation of targets and the distance between the point of execution and the target. 40 male children ages 9 to 10 years and 40 male students ages 18 to 19 years were divided into eight equal groups according to age and training group (No Specific target, 1 target, 4 targets, and 5 targets). After 18 practice sessions over 40 days and similar teaching, the participants performed 60 transfer trials after a rest of 72 hours. The experimental design used was a 2 (age) x 4 (practice group) x 6 (block of trials) with repeated measures on the last factor for absolute and constant error scores. The analysis of variance showed that the children performed more poorly than the adolescents, and variable practice yielded better performance than constant and specialized practice. The results support the variability of practice hypothesis and indicate that the effect of variable and specialized practic was related to age of the groups of trainees. PMID- 16383092 TI - Awareness of the false memory manipulation and false recall for people's names as critical lures in the Deese-Roediger-McDermott paradigm. AB - In the present study, people's names were used as Critical Lures (central concept not on the lists) in the Deese-Roediger-McDermott (DRM) paradigm for studying false recall. Following Mukai, the transparency of false memory manipulation in the paradigm was manipulated to explore its influence on false recall. 80 volunteers (age range 17-30 years, M age = 20.9 yr., SD = 1.6) took part. Analyses showed that making the critical lure inconspicuous did not significantly increase false recall. However, the proportion of falsely recalled Critical Lures was significantly lower and the proportion of critical lures produced on a postrecall test (asking participants to report items that they had thought of but did not recall) was significantly higher when they spontaneously realized the nature of false memory manipulation. However, there was no effect on veridical recall of study items. These results suggest that participants strategically avoided reporting false recall for people's names as Critical Lures even without forewarning about false memory. PMID- 16383093 TI - Effects on reading performance of indention patterns in a Chinese sign. AB - This study investigated the effects of the use of indention in a public Chinese sign which can provide the reader with cues for the appropriate eye direction for reading a sign which facilitates reading. The between-subjects design had repeated observations on the indention design. The right or left column was indented in a Chinese sign of only two columns. 144 college students in Taiwan who served as subjects were divided into two groups by their reading comprehension and shown stimuli in a tachistoscope. Analysis showed that an indented design significantly reduced reading time but did not change accuracy rate. This result was consistent with the results of Duchnicky and Kolers' 1983 study and indicated that a reader can maintain accuracy while reducing reading time when reading a text designed with different kinds of indention layout. These results might aid designers of Chinese warning signs, especially for readers with lower reading scores. PMID- 16383094 TI - Note regarding the word 'behavior' in glossaries of introductory textbooks, dictionaries, and encyclopedias devoted to psychology. AB - Glossaries of introductory textbooks in psychology, biology, and animal behavior were surveyed to find whether they induded the word 'behavior'. In addition to texts, encyclopedias and dictionaries devoted to the study of behavior were also surveyed. Of the 138 tests sampled across all three fields, only 38 (27%) included the term 'behavior' in their glossaries. Of the 15 encyclopedias and dictionaries surveyed, only 5 defined 'behavior'. To assess whether the term 'behavior' has disappeared from textbook glossaries or whether it has usually been absent, we sampled 23 introductory psychology texts written from 1886 to 1958. Only two texts contained glossaries, and the word 'behavior' was defined in both. An informal survey was conducted of students enrolled in introductory classes in psychology, biology, and animal behavior to provide data on the consistency of definitions. Students were asked to "define the word 'behavior'." Analysis indicated the definition was dependent upon the course. We suggest that future introductory textbook authors and editors of psychology-based dictionaries and encyclopedias include 'behavior' in their glossaries. PMID- 16383095 TI - Rapid plastic changes of human primary motor cortex with repetitive motor practice and transcranial magnetic stimulation. AB - Excitability changes of human primary motor cortex are assumed to be associated with motor learning processes. To examine motor behavioral and neural mechanisms in these processes, the adaptive motor learning processes of the index finger abduction were investigated using motor evoked potential (MEP) elicited from the first dorsal interosseous and extensor carpi radialis muscles. Practice effects were examined on changes of MEP amplitudes elicited from these muscles during motor imagery. Given general consensus that the MEP amplitude change during motor imagery is a useful parameter reflecting changes in excitability of the human primary motor cortex, the present results, that MEP amplitudes of both muscles increased with repeated practice by the index finger abduction and that magnitudes of MEP amplitudes of both muscles (motor learning curves) were clearly different, suggested that participation of the muscles performing the index finger abduction gradually changed with practice. Short-term plastic changes of human primary motor cortex occur with repetitive practice and such adaptive change in human primary motor cortex is expressed in human voluntary movement that becomes more automatic. PMID- 16383096 TI - Evaluation of a planned behavior theory-based intervention programme to promote healthy eating. AB - The objective of the study was to test the effectiveness of an intervention program based on the theoretical framework of the Theory of Planned Behavior, with the addition of attitude strength and role identity. The aim was to alter adolescents' healthy eating attitudes and behaviour. In the sample were 335 high school students, who were divided into intervention and control groups. The intervention lasted 12 weeks and included posters and lectures promoting healthy eating. The measures included a questionnaire assessing the hypothesis and a food frequency questionnaire which measured eating habits. Analysis showed the intervention was effective in proving attitudes toward healthy eating and attitude strength, intention, perceived behavioral control, and healthy eating behaviour, but not effective in predicting subjective norms and role identity. Results provide evidence that intervention changed attitudes toward a behavior in a school setting. PMID- 16383097 TI - Comparing performance by parents and offspring on a rapid tapping task. AB - The purpose of this study was to identify the relationships between 42 parents' and their offsprings' performance on a rapid tapping task. It was expected that genetic predispositions would gradually limit children's performance on a tapping task as they matured chronologically. Four different age groups of children and their parents performed the tapping task on different sizes of boards. Repeated measures analysis of variance indicated that, while parents' mean performance was generally higher than that of their children (F = 32.89, p < .001), the difference was affected by the children's age, which hand was used, and the board size. Spearman rank correlation scores between parents' and children's overall performances gradually increased across age groups (p = -.29, -.10, .47, and .40, respectively, from younger to older age groups). This finding suggests that the potential of youngsters' future performance may be predicted with greater certainty by observing their parents' present performance. This may improve the predictive power of planned programs. PMID- 16383098 TI - Dream recall and sleep duration: state or trait factor. AB - Previous research regarding the relationship between habitual sleep duration and frequency of dream recall yielded conflicting results, whereas experimental manipulation of sleep duration showed a significant effect on dream recall. In this study analyzing both effects simultaneously, the findings from diaries kept by the 196 participants over 28 to 111 days indicated that intra-individual fluctuations in sleep duration were significantly related to dream recall whereas interindividual differences in dream recall were not associated with differences in sleep duration. Sleep inertia might be an explanation for this finding. PMID- 16383099 TI - Effects of participants' sex and targets' perceived need on supermarket helping behavior. AB - A field experiment was focused on whether participants' sex and targets' perceived need influenced helping behavior. Confederates approached 332 (166 women, 166 men) same-sex participants in a supermarket and asked for 25 cents to help purchase one of three randomly assigned food items: milk which was defined as a high-need item, frozen cookie dough which served as a low-need item, or alcohol, which was a low-need item with negative social connotations. The dependent variable was whether a participant provided help. Participants' sex was not associated with helping behavior as equal proportions of men and women provided assistance to the confederate; however, perceived need strongly influenced whether the confederate received help. Specifically, the high-need item produced more helping behavior than did either of the low-need items, and the socially acceptable low-need item of cookie dough produced more helping behavior than the socially unacceptable low-need item of alcohol. This may be interpreted as showing that what one buys and how deserving of help one appears to be influence whether one is helped by others. PMID- 16383100 TI - Encoding richness of self-generated elaboration and spacing effects on incidental memory. AB - The present study investigated encoding variability in self-generated elaboration on incidental memory as a function of the type of presentation which was either massed or spaced. The subjects generated different answers to a "why" question for the first and the second presentations of a target sentence in a self generated elaboration condition. In an experimenter-provided elaboration condition they then rated the appropriateness of the different answers provided by the experimenter for the first and second presentations. This procedure was followed by two free recall tests, one of which was immediate and the other delayed. A self-generated elaboration effect was observed in both the spaced and the massed presentations. These results indicated that the self-generated elaboration effect was facilitated, even in the massed presentation because the different answers to the first and the second presentations led to a richer encoding of each target. PMID- 16383101 TI - Preliminary report: psychological assessment of Greek women with diabetes during pregnancy. AB - 23 Greek pregnant women with type 1 diabetes had a higher mean score on the Maudsley Obsessive-Compulsive Inventory in the second and third trimesters of pregnancy than 13 women with gestational diabetes. Long-term changes in the lifestyle of the former may apparently lead to this higher mean. For those with gestational diabetes, higher scores of alexithymia were correlated with slightly worse glycemic control. PMID- 16383102 TI - Evaluation and classification of types of Chinese handwriting deficits in elementary schoolchildren. AB - The handwriting problems of elementary schoolchildren were investigated in this study. A questionnaire for evaluation of Chinese handwriting was developed and then administered to 165 children with handwriting deficits. Factor analysis indicated that the questionnaire has six major dimensions, including construction of characters, accuracy, developmental delay, pencil grip, gross movement, and emotional reaction. These six dimensions were utilized to search for subtypes of handwriting deficits. By further sampling, another 209 children were administered the handwriting evaluation questionnaire to classify their handwriting deficits. According to the severity of deficits on different dimensions of the questionnaire, K-means clustering was utilized to classify the children into four subtypes of mild, severe, motor-originated, and academic learning dysfunction subtypes. The classification by subtype might be helpful in planning interventions for children with handwriting deficits. PMID- 16383103 TI - Treating Parkinsonian speech: development of a feedback rating scale using a case study. AB - The speech of people with Parkinson's disease is often unintelligible because the speaker has limited volume and imprecisely articulated speech. The Lee Silverman Voice Treatment addresses volume by requiring a patient to exert extreme effort to adduct the vocal folds and increase volume. Little attention, however, is paid to articulation. Such patients often have perceptual difficulties which prevent them from monitoring their own volume and speech. A case study presents a method for improving volume and articulation of speech of a patient with Parkinson's disease by focusing on perceptual aspects of speech. PMID- 16383104 TI - Relations of perceived physical self-efficacy and motivational responses toward physical activity by urban high school students. AB - To be effective in promoting physical activity among urban, minority adolescents, the factors and psychological processes that motivate them to engage in and maintain a physically active lifestyle should be examined. The relation of physical self-efficacy and motivational responses toward physical activity in 46 urban minority adolescents was explored. As hypothesized, there were significant positive relationships among Percieved Physical Ability, Physical Self presentation Confidence, Effort, and Enjoyment (coefficients ranged from .29 to .80), suggesting that participants who had higher perceived physical ability were likely to report higher perceptions of self-presentation, more enjoyment of physical activity, and harder work in physical activity. These results indicate specific relationships among Effort, Enjoyment, Perceived Physical Ability, and Physical Self-presentation Confidence in this sample. Physical self-efficacy appears to be a stronger predictor of motivational responses in physical activity. Practical implications for physical educators include incorporating strategies known to develop self-efficacy, such as mastery experiences involving successive trials of increasing difficulty, self-observation, external feedback, peer modeling, and verbal persuasion. PMID- 16383105 TI - Incremental exercise in dynamic visual acuity. AB - As research is limited the purpose of this study was to investigate dynamic visual acuity while cycling at different exercise loads. Accuracy of dynamic visual acuity scores of 20 college-age participants was analyzed by a repeated measures analysis of variance which indicated improved dynamic visual acuity during cycling as the intensity of exercise increased. A retention test conducted two days after testing yielded evidence of trainability of dynamic visual acuity. In the discussion findings were compared to other visual functions associated with exercise. PMID- 16383106 TI - Use of rest breaks and accidents by professional drivers. AB - This study examined the effect of rest breaks on traffic accidents by 720 randomly selected professional drivers who completed a postal questionnaire. Drivers involved in an accident preferred to enjoy their breaks at rest places meant for professional drivers, whereas accident-free drivers chose more freely the timing of their breaks during driving. PMID- 16383107 TI - Comparison of spectrographic records of two syllables pronounced from scripts in hiragana and romaji by students with different familiarity with English. AB - The same syllables /mu/ and /ra/ written in Japanese hiragana and romaji given on a standard speeded naming task elicited phonetically or acoustically different responses in a syllabic hiragana condition and a romaji condition. The participants were two groups of Japanese college students (ns = 15 and 16) with different familiarity with English as a second language. The results suggested that the phonetic reality of syllables represented in these scripts can differ, depending on the interaction between the kind of script and speakers' orthographic familiarity. PMID- 16383108 TI - Burden of intractable epilepsy. PMID- 16383109 TI - Defining intractable epilepsy. AB - There is no single definition of medically refractory epilepsy, and different definitions may be appropriate depending on the form of epilepsy or on the purposes for which the definition is being used. The natural history of the specific form of epilepsy must be accurately characterized to facilitate a meaningful definition of refractory for that form of epilepsy and thereby allow identification as soon as possible in the course of the disorder. Long-term follow-up after appearance of intractable epilepsy (as determined by a variety of definitions) would provide helpful information necessary for refining meaningful definitions of intractability. PMID- 16383110 TI - Outcome measures in intractable epilepsy. PMID- 16383111 TI - Focal seizures: intractability and semiology. PMID- 16383112 TI - The syndrome of mesial temporal lobe epilepsy with hippocampal sclerosis: clinical features and differential diagnosis. PMID- 16383113 TI - Clinical aspects of temporal/limbic epilepsy and their relationships to intractability. PMID- 16383114 TI - Clinical electrophysiology factors indicative of intractability in patients with temporal lobe epilepsy. PMID- 16383115 TI - Epilepsy-based changes in hippocampal excitability: causes and effects. PMID- 16383116 TI - Selective degeneration and synaptic reorganization of hippocampal interneurons in a chronic model of temporal lobe epilepsy. AB - Synaptic properties and connectivity of GABAergic inhibitory interneurons are modified in CA1 hippocampus of the KA model of epilepsy (Fig. 1). These changes affect interneurons that target dendritic areas of principal cells and have important consequences for network activity and hyperexcitability. Some of these changes contribute to hyperexcitability, while others likely develop to compensate for the hyperexcitability of the network. However, some of these "compensatory" changes might also paradoxically contribute to hyperexcitability and epileptogenesis. PMID- 16383117 TI - Parahippocampal networks, intractability, and the chronic epilepsy of kindling. AB - Clearly, the root cause of intractability in epilepsy is currently unknown. Whereas the aforementioned findings may shed light on putative underpinnings, they are by no means an exhaustive list of possibilities. However, new and more effective animal models are continually being created or discovered that take into account genetic predisposition for seizure. At the moment, amygdala kindling appears to be the best choice of the intact animal models. In this vein, the genetically predisposed seizure-prone (Fast kindling) and seizure-resistant (Slow kindling) strains may help speak to many important remaining questions in human epilepsy. Hopefully, these models, to some degree, target correct human subpopulations that are prone or resistant to epilepsy and, when used appropriately, could expedite epilepsy research and future discoveries leading to pharmacoresistance and intractability. PMID- 16383118 TI - Novel mechanisms underlying drug resistance in temporal lobe epilepsy. PMID- 16383119 TI - Failure of surgery for temporal lobe epilepsy: a review of selected aspects. PMID- 16383120 TI - Neocortical epilepsy: aspects distinguishing intractable from benign neocortical epilepsies. PMID- 16383121 TI - Intractable epilepsies of the neocortex: basic mechanisms. PMID- 16383122 TI - Simultaneous electroencephalogram-functional magnetic resonance imaging in neocortical epilepsies. PMID- 16383123 TI - What intractability information is there in the type of generalized seizure? PMID- 16383124 TI - Neuronal substrates of spike-wave seizures and hypsarrhythmia in corticothalamic systems. PMID- 16383125 TI - Novel strategies for the development of animal models of refractory epilepsy. PMID- 16383126 TI - Participation of cortical glial cells to the genesis of spike-wave seizures. PMID- 16383127 TI - Status epilepticus and refractory status epilepticus: introductory and summary statements. PMID- 16383128 TI - Basic mechanisms in status epilepticus: role of calcium in neuronal injury and the induction of epileptogenesis. PMID- 16383129 TI - Epidemiology and clinical presentation of status epilepticus. PMID- 16383130 TI - Status epilepticus and brain damage: pathology and pathophysiology. PMID- 16383131 TI - Nonconvulsive seizures and electroencephalogram monitoring in the intensive care unit. PMID- 16383132 TI - Status epilepticus in pediatric practice: neonate to adolescent. AB - SE in the pediatric age group presents challenges in diagnosis and management. There is need for renewed consensus on the temporal definition of SE, both clinical and electrographic. SE in children exhibits an age-dependent vulnerability, with genetic predisposition and etiology as determinants of susceptibility. Nonepileptic phenomena may mimic SE. Clinical and electrographic SE in neonates are relatively rare, while serial (clinical and electrographic) and repetitive seizures are more common. Neurometabolic disease, chromosomal disorders, and abnormalities of cortical development are important etiological considerations. Abrupt discontinuation of or an aberrant response to AEDs can also precipitate SE. Metabolic perturbations and toxins can further aggravate the situation. Clinical and experimental data suggest that the longer a seizure lasts, the more difficult it becomes to control, and that seizures can have immediate and long-term adverse consequences on the immature and developing brain. Hence, treatment (usually with a benzodiazepine) should be started early in the clinical course. A trial of pyridoxine, biotin, or folinic acid should be considered in the appropriate clinical setting (e.g., neonates or young infants, in particular). Phenytoin/fosphenytoin and phenobarbital remain important treatment options. Pentobarbital and midazolam are preferred choices in the management of RSE. Once metabolic causes are excluded, children with RSE should be evaluated for surgical treatment early in the clinical course. Clinical guidelines based on best available evidence have to be periodically reviewed. The clinical consequences and management of electrographic SE, especially in the neonate, have to be addressed. Guidelines for continuous (video) EEG monitoring are needed to facilitate this task. AEDs that do not have an adverse effect on the developing brain have to be developed. Our review suggests a continuing need for prospective studies into all aspects of SE in the pediatric age group. PMID- 16383133 TI - Current approach to the diagnosis and treatment of refractory status epilepticus. PMID- 16383134 TI - Magnetic resonance imaging sequences for detection of lesions in refractory epilepsy. AB - Typical sequences in MR imaging of epilepsy been described, in addition to a brief overview of imaging abnormalities that may be seen. The interested reader is referred to a recent publication, Neuroimaging Clinics of North America on Epilepsy, which goes into considerable detail on imaging, including MRI and other modalities. PMID- 16383135 TI - Magnetic resonance imaging in intractable epilepsy: focus on structural image analysis. PMID- 16383136 TI - High-resolution magnetic resonance imaging (MRI), surface coil MRI, and magnetoencephalography. AB - The high-resolution, phased array, or surface coil techniques offer promise of improvements in diagnosis of cortical abnormalities. Further prospective studies in larger numbers of patients will be needed to determine the exact improvement, which might be expected from experienced image interpreters. Combining high resolution imaging with MEG would seem to be the best noninvasive way of accurately localizing seizure foci, and any potential anatomic derangements, currently available to us. PMID- 16383137 TI - Magnetic resonance spectroscopic techniques for localizing epileptogenic zones in refractory partial epilepsy. PMID- 16383138 TI - Animal models of epilepsy and progressive effects of seizures. PMID- 16383139 TI - Mechanisms that might underlie progression of the epilepsies and how to potentially alter them. PMID- 16383140 TI - Clinical evidence that epilepsy is not a progressive disorder with special emphasis on epilepsy syndromes that do not progress. PMID- 16383141 TI - Clinical evidence that epilepsy is a progressive disorder with special emphasis on epilepsy syndromes that do progress. PMID- 16383142 TI - Psychosocial comorbidity in epilepsy. PMID- 16383143 TI - Behavioral comorbidities: attention deficit hyperactivity disorder--human and animal studies. PMID- 16383144 TI - Medical comorbidity in intractable epilepsy. PMID- 16383145 TI - Intractable epilepsy, hormonal and reproductive problems: human and animal studies. AB - Both women and men with epilepsy run significant risks of reproductive dysfunction. These problems are seldom addressed, however, either by the medical profession or by lay organizations. Many physicians are not even aware of these health risks. Increased awareness of--and treatment of--these problems are important goals for the future. PMID- 16383146 TI - Neuropsychological profiles. PMID- 16383147 TI - Intractable epilepsy and patterns of psychiatric comorbidity. PMID- 16383148 TI - Psychological treatment in intractable epilepsy: approaches, evidence, and future directions. PMID- 16383149 TI - Monogenic epilepsies in humans: molecular mechanisms and relevance for the study of intractable epilepsy. PMID- 16383150 TI - Polygenic epilepsy. AB - Current perspective on the etiology of common forms of epilepsy reflects clinical and preclinical data that document a complex determinism involving multiple gene variations and environmental influences. This multifactorial architecture precludes effective use of conventional types of genetic analysis and requires application of alternative methods. Presently, such methods are based on candidate gene studies, and it is likely that these approaches will continue to be useful in the near term. Long-range strategies will combine traditional linkage methods with single nucleotide polymorphism gene maps and will seek to collect data on a genomic scale in the search for gene variation that influences both the development of epilepsy and the response to antiepileptic medication. Such studies will require large numbers of patients and controls and also valid, standardized clinical criteria for parsing patients into phenotypic categories. Animal models will continue to provide clues regarding potentially important candidate genes and can also be used in a reverse-translational manner to confirm and study the effects of gene variation that is identified in humans. In the end, genetic research approaches to common forms of human epilepsy will enhance understanding of underlying neurobiological mechanisms, and through this understanding will come new treatment options, new molecular markers to improve current diagnostic criteria, and new genetic tests for predicting drug efficacy in patients with epilepsy. PMID- 16383151 TI - On the road to tractability: the current biochemical understanding of progressive myoclonus epilepsies. PMID- 16383152 TI - Major targets and mechanisms of antiepileptic drugs and major reasons for failure. PMID- 16383153 TI - Designing future drugs for the treatment of intractable epilepsy. PMID- 16383154 TI - The definition and prediction of intractable epilepsy in children. PMID- 16383155 TI - Diseases and syndromes associated with intractable epilepsy. PMID- 16383156 TI - Management of intractable epilepsy in infancy and childhood. PMID- 16383157 TI - The spectrum of intractable epilepsy: a proposed scale. PMID- 16383158 TI - Differential diagnostic considerations in patients with intractable epilepsy. PMID- 16383159 TI - Investigations: how the data affect management. PMID- 16383160 TI - Medical management of intractable epilepsy. PMID- 16383161 TI - Assessment of patients with intractable epilepsy for surgery. PMID- 16383162 TI - Palliative surgery for intractable epilepsy. PMID- 16383163 TI - Evaluating surgery effectiveness. PMID- 16383164 TI - Deep brain stimulation and cortical stimulation in the treatment of epilepsy. PMID- 16383165 TI - Source monitoring in eyewitness memory: implicit associations, suggestions, and episodic traces. AB - Both the distinctiveness heuristic and discrepancy detection hypotheses were investigated by independently manipulating both schema consistency and incidental suggestion in an eyewitness memory paradigm. A sequence of slides was shown, followed by a postevent questionnaire that contained both schema-typical and schema-atypical information. Fifteen minutes later, a source-monitoring task was administered. In Experiment 1, the proportion of source misattribution errors was greater for schema-typical items than for schema-atypical items, and the proportion of errors on suggested items was greater than that on control items. Suggestion affected schema-typical and schema-atypical items equally, providing no support for the predictions of either hypothesis. In Experiment 2, the interval between the questionnaire and the source test was manipulated. The results of Experiment 1 were replicated under the short delay, whereas the proportion of errors increased under the long delay. An associative network model involving two types of episodic traces was used to account for the results. PMID- 16383166 TI - How eyewitnesses resist misinformation: social postwarnings and the monitoring of memory characteristics. AB - Previous findings have been equivocal as to whether the postevent misinformation effect on eyewitness memory is reduced by warnings presented after the misinformation (postwarnings). In the present research, social postwarnings, which characterize the postevent source as a low-credibility individual, diminished the misinformation effect in both cued recall and recognition tests. Discrediting the source as being either untrustworthy or incompetent was effective (Experiment 1). Also, postwarned participants rated reality characteristics of their memories more accurately than did participants receiving no or high-credibility information about the postevent source (Experiment 2). A social postwarning yielded the same results as an explicit source-monitoring appeal and led to longer response times for postevent items, relative to a no warning condition (Experiments 3 and 4). The findings suggest that the reduced misinformation effect was due to more thorough monitoring of memory characteristics by postwarned participants, rather than to a stricter response criterion or to enhanced event memory. PMID- 16383167 TI - Eyewitness decisions in simultaneous and sequential lineups: a dual-process signal detection theory analysis. AB - Many eyewitness researchers have argued for the application of a sequential alternative to the traditional simultaneous lineup, given its role in decreasing false identifications of innocent suspects (sequential superiority effect). However, Ebbesen and Flowe (2002) have recently noted that sequential lineups may merely bring about a shift in response criterion, having no effect on discrimination accuracy. We explored this claim, using a method that allows signal detection theory measures to be collected from eyewitnesses. In three experiments, lineup type was factorially combined with conditions expected to influence response criterion and/or discrimination accuracy. Results were consistent with signal detection theory predictions, including that of a conservative criterion shift with the sequential presentation of lineups. In a fourth experiment, we explored the phenomenological basis for the criterion shift, using the remember-know-guess procedure. In accord with previous research, the criterion shift in sequential lineups was associated with a reduction in familiarity-based responding. It is proposed that the relative similarity between lineup members may create a context in which fluency-based processing is facilitated to a greater extent when lineup members are presented simultaneously. PMID- 16383168 TI - Remembering the news: modeling retention data from a study with 14,000 participants. AB - A retention study is presented in which participants answered questions about news events, with a retention interval that varied within participants between 1 day and 2 years. The study involved more than 14,000 participants and around 500,000 data points. The data were analyzed separately for participants who answered questions in Dutch or in English, providing an opportunity for replication. We fitted models of varying complexity to the data in order to test several hypotheses concerning retention. Evidence for an asymptote in retention was found in only one data set, and participants with greater media exposure displayed a higher degree of learning but no difference in forgetting. Thus, forgetting was independent of initial learning. Older adults were found to have forgetting curves similar to those of younger adults. PMID- 16383169 TI - Mechanisms of autobiographical memory retrieval in younger and older adults. AB - In this study, retrieval of autobiographical memories was compared within or without the context of a lifetime period and between younger and older adults. The results of two experiments demonstrated that memories were more accessible for younger and older adults when they were retrieved within a time period context than without such context. However, when the retrieval context was more constrained and limited to cue word memories, the benefit of retrieval context disappeared. The results also suggested that as compared with younger adults, older adults were more likely to selectively retain memories with distinctive characteristics, such as being self-relevant and emotionally intense, particularly when remote memories were involved. Mechanisms of autobiographical memory retrieval in younger and older adults. PMID- 16383170 TI - When and why is ease of retrieval informative? AB - In two experiments, we examined when and why ease of retrieval of information from memory affects behavioral frequency and attitudinal judgments. Overall, the results suggest that when the subjective experience of ease of retrieval is consistent with the expected ease of retrieval, the content of the information retrieved is used to make judgments. However, when there is a discrepancy between experienced and expected ease of retrieval, the subjective experience of ease of retrieval is used to make judgments. Ease of retrieval is more informative when the discrepancy between experienced and expected ease of retrieval cannot be attributed to task contingencies; when it can, ease of retrieval ceases to be informative. PMID- 16383171 TI - A comparative analysis of serial and free recall. AB - Multitrial free and serial recall tasks differ both in recall instruction and in presentation order across trials. Waugh (1961) compared these paradigms with an intermediate condition: free recall with constant presentation order. She concluded that differences between free and serial recall were due only to recall instructions, and not to presentation order. The present study reevaluated the relation between free and serial recall, using Waugh's three conditions. By examining recall transitions and the organization of information retained across trials, we conclude that presentation order is an important factor, causing participants to exhibit the same temporal associations in serial recall and in free recall with constant presentation order. PMID- 16383172 TI - Is memory for stimulus magnitude Bayesian? AB - This study was designed to determine whether memory for stimulus values is a Bayesian weighting of the magnitude of a stimulus and the central tendency of an exemplar's category (Huttenlocher, Hedges, & Vevea, 2000). In five experiments, participants reproduced the remembered size of a geometric figure drawn from one of two categories whose means for size differed. Reproductions were biased toward the mean of the combined distribution rather than the mean of either category. Reproductions were also influenced by the size of the stimulus on the preceding trial. Neither of these results is entirely consistent with the view that recollections are partially constructed from a consideration of the long-run probabilities established by category membership. PMID- 16383173 TI - Does the order of head noun and modifier explain response times in conceptual combination? AB - We describe a speeded sensibility judgment experiment in which noun-noun combinations in the Indonesian language were used that parallels Gagne and Shoben's (1997) study of combinations in the English language. Like English, Indonesian is read from left to right and contains noun-noun combinations that are formed by juxtaposing the nouns. However, unlike in English, the order of the modifier and the head noun is reversed. This difference between English and Indonesian combinations allowed us to assess whether sensibility judgments of combinations are affected primarily by the left-right order of the nouns or by different functional roles of the nouns (i.e., modifier vs. head noun). As in Gagne and Shoben's study, the modifier's relation frequency contributed significantly to predicting sensibility judgment times in a regression analysis, but the head noun's relation frequency did not. We discuss the implications of this finding for models of conceptual combination. PMID- 16383174 TI - Prosody and lemma selection. AB - In three picture-naming experiments, we examined the effect of prosodic context on the synonyms people use to name pictures in Mandarin Chinese. This was done without time pressure. The results showed that when monosyllabic and bisyllabic synonyms (e.g., hen/chicken) were embedded in a context of pictures with either bisyllabic or trisyllabic names, participants gave bisyllabic responses to the synonyms more often than they did in a condition without such a context. The difference was very similar in magnitude in both the bisyllabic and trisyllabic contextual conditions. These results suggest that people are biased toward using synonyms that have numbers of syllables equal or similar to those of the prosodic context. If it is assumed that prosodic effects originate at a stage of processing beyond the lemma level, then this suggests either that multiple phonological forms of synonyms can be activated or that there is feedback from prosodic processing that influences lemma selection. PMID- 16383175 TI - Both sides get the point: hemispheric sensitivities to sentential constraint. AB - Behavioral studies have consistently reported striking differences in the impact of sentence-level information on the processing of words presented in the right (RVF) versus the left (LVF) visual field, with context effects apparent only for RVF items. The consistent lack of such effects in the LVF has been taken to mean that right hemisphere language comprehension is largely insensitive to message level meaning. We used the functional specificity afforded by event-related potential measures to assess this claim. Target words completing strongly and weakly constraining sentence contexts, in which constraint arose at the sentence level rather than from lexical associations, were presented laterally in the LVF or RVF. Increased constraint significantly reduced N400 amplitudes with presentation in both VFs, with no differences in the timing or amplitude of these effects. These results are inconsistent with the view that the VF asymmetries found in behavioral measures reflect differential hemispheric capacities at the level of semantic analysis and integration, although VF-based differences on earlier components (P2) suggest asymmetries in the impact of sentential context on perceptual aspects of word processing in the two hemispheres. PMID- 16383176 TI - The perception of handshapes in American sign language. AB - Despite the constantly varying stream of sensory information that surrounds us, we humans can discern the small building blocks of words that constitute language (phonetic forms) and perceive them categorically (categorical perception, CP). Decades of controversy have prevailed regarding what is at the heart of CP, with many arguing that it is due to domain-general perceptual processing and others that it is determined by the existence of domain-specific linguistic processing. What is most key: perceptual or linguistic patterns? Here, we study whether CP occurs with soundless handshapes that are nonetheless phonetic in American Sign Language (ASL), in signers and nonsigners. Using innovative methods and analyses of identification and, crucially, discrimination tasks, we found that both groups separated the soundless handshapes into two classes perceptually but that only the ASL signers exhibited linguistic CR These findings suggest that CP of linguistic stimuli is based on linguistic categorization, rather than on purely perceptual categorization. PMID- 16383177 TI - Age of acquisition effects in the semantic processing of pictures. AB - In two experiments, we investigated the role of age of acquisition (AoA) in the categorizing of objects in semantic tasks that do not require access to the object names. In both a found inside or outside the house (Experiment 1A) and a smaller or larger than a loaf of bread (Experiment 2A) classification task, objects with earlier-acquired names were categorized more quickly than those with later-acquired names. Experiments 1B and 2B also showed AoA effects on object naming times for the same pictures. We conclude that AoA operates within the identification process in a fashion not simply restricted to name retrieval. These effects may be better explained in terms of the connectionist model proposed by Ellis and Lambon Ralph (2000) or by accounts that locate AoA within the semantic system (e.g., Brysbaert, Van Wijnendaele, & De Deyne, 2000; van Loon Vervoorn, 1989). PMID- 16383178 TI - Symbolic distance affects two processing loci in the number comparison task. AB - A dual-task procedure was used to investigate the attentional requirements of number processing. The results show that (1) numeric information in Task 2 can be retrieved in parallel with capacity-demanding processing in Task 1 but (2) comparing two quantities requires central capacity, which is depleted by switching from one task to another. This finding resolves an apparent discrepancy in the literature, in which digit magnitude information has not been retrieved in parallel with a second task (Logan & Schulkind, 2000), despite repeated demonstrations that this information is retrieved autonomously, even when it is deleterious to performance (Henik & Tzelgov, 1982). A model is proposed to reconcile existing findings with the new ones revealed in the present investigation. PMID- 16383179 TI - Strategy use, the development of automaticity, and working memory involvement in complex multiplication. AB - Participants practiced a set of complex multiplication problems (e.g., 3 x 18) in a pre-/postpractice design. Before, during, and after practice, the participants gave self-reports of problem-solving strategies. At prepractice, the most common strategy was a mental version of the standard multidigit algorithm, and dual tasks revealed that working memory load was high and heavier for problems solved via nonretrieval strategies. After practice, retrieval was used almost exclusively, and participant variability, automaticity level of problems (proportion of trials on which retrieval was used over the entire experiment), and error rates were significant predictors of problem-solving latencies. Practice reduced working memory involvement to minimal levels, and there was no relationship between automaticity level and working memory load. The commonalities between the present findings and findings related to automaticity development in simple arithmetic are discussed. PMID- 16383180 TI - Putting babies "back to sleep": can we do better? AB - The American Academy of Pediatrics first recommended in 1992 that infants be placed on their backs for sleep to reduce the risk for sudden infant death syndrome (SIDS). Since that time, there has been a national drop in the incidence of SIDS of more than 40 percent. Unfortunately, many parents and other caregivers are still receiving inconsistent information and observing varying practices regarding infant sleep position. This article emphasizes the role of the health care professional in both teaching and modeling these potentially life-saving practices consistently and unambiguously. Available literature is reviewed regarding attitudes and beliefs about infant sleep positioning, and specific concerns are addressed in order to allow for better tailoring of educational programs. PMID- 16383181 TI - Intravenous immunoglobulin in the management of hemolytic disease of the newborn. AB - Jaundice caused by hemolysis continues to challenge practitioners caring for infants in the NICU. Bilirubin levels can rise quickly in the first days of life, and interventions must be prompt to prevent side effects related to hyperbilirubinemia. Conventional treatments such as hydration and phototherapy are common, but new studies suggest that use of intravenous immunoglobin (IVIG) as an additional treatment may prevent the need for exchange transfusion in some babies. This article presents a case study of an infant with blood-type incompatibility treated successfully with multiple doses of IVIG, discusses the pathophysiology and clinical presentation of hemolytic jaundice, and reviews current management strategies for this disease. PMID- 16383182 TI - A concept analysis of family-centered care in the NICU. AB - Family-centered care (FCC) has been implemented in many NICUs throughout the U.S. It is valuable in helping families whose infants require hospitalization cope with the stress, fear, and altered parenting roles that may accompany their child's condition and hospitalization. To employ such a significant philosophy of care, nurses must understand what the FCC concept signifies. A concept analysis can often aid understanding of abstract ideas such as FCC. This article utilizes a 1995 framework for concept analysis to clarify the meaning of FCC for the neonatal nurse. Incorporating FCC into daily professional practice can enable nurses to improve the emotional and physical well-being of each family they encounter. PMID- 16383183 TI - Elevated sound levels within a busy NICU. AB - PURPOSE: Elevated sound levels in the NICU may contribute to undesirable physiologic and behavioral effects in preterm infants. This study describes sound levels in a busy NICU in the southeastern U.S. and compares the findings with recommended NICU noise level standards. DESIGN: NICU sound levels were recorded continuously at nine different locations within the NICU. Hourly measurements of loudness equivalent (Leq) sound level, sound level exceeded 10 percent of the time (L10), and maximum sound level (Lmax) were determined. SAMPLE: Sound levels were sampled from nine different locations within the NICU. MAIN OUTCOME VARIABLE: Sound levels are described using the hourly, A-weighted Leq, L10, and Lmax. RESULTS: The overall average hourly Leq (M = 60.44 dB, range = 55-68 dB), L10 (M = 59.26 dB, range = 55-66 dB), and Lmax (M = 78.39, range = 69-93 dB) were often above the recommended sound levels (hourly Leq <50 dB, L10 <55 dB, and 1 second Lmax <70 dB). In addition, certain times of day, such as 6-7 AM and 10 AM 12 noon, were noisier than other times of day. PMID- 16383184 TI - Creating a winning team: lessons from football. AB - There are tasks best done on an individual basis when caring for a neonate, but the ultimate outcome for infants and their families results from a team effort. Incorporating ten strategies drawn from football can help the NICU manager create and foster effective teamwork. PMID- 16383185 TI - Findings from the National Institute of Nursing research related to neonatal care: 2005 update. AB - This annotated bibliography from the National Institute of Nursing Research (NINR) presents recent findings on a variety of studies related to prenatal conditions and the benefits of prenatal care, advances in our understanding about infant care and family issues in the newborn intensive care unit (NICU), and the importance of clinicians in supporting and communicating with the family. By sharing this bibliography, we hope to increase the awareness of these valuable research findings within the nursing community and support the continued development of evidence-based practice in antenatal, postpartum, and neonatal care. PMID- 16383186 TI - [Contemporary programme of pharmaceutical studies in Germany]. AB - In the year 2002, both the Czech and Slovak Faculties of Pharmacy celebrated the fiftieth anniversary of the existence of four-year and later five-year university courses of pharmaceutical studies, the contents of which were being continuously modified in order to comply with principal trends in the field of pharmacy. A new programme of pharmaceutical studies which has just started at German universities can serve us as a certain example and provide comparison, or inspiration. PMID- 16383187 TI - [Logistics in the pharmaceutical service]. AB - The conception of the field of pharmaceutical service defines pharmaceutical service as the basic part of pharmacy, the principal task of which is to provide pharmaceutical care as an inseparable part of providing health care. It represents a set of professional activities of the pharmacist oriented to securing human and veterinary pharmaceutical products and health care products and to optimising effective, safe and quality pharmacotherapy. Technically, pharmaceutical service is an applied discipline, as it makes use of knowledge gained in other pharmaceutical, medical, psychological, social, and economic sciences. Because of its interdisciplinary character it is necessary to extend the theory of pharmaceutical service in such a way so that it may reflect all aspects of its sphere of activity. One of the possibilities is to define the pharmacy premises as an independent functional unit which operates on the basis of valid legal standards in such a way that on the one hand it secures the provision of health (pharmaceutical) care, and on the other hand it maintains its cost-effectiveness. To keep the quality of care of the patient and the economic aspect in balance, it is necessary to define the term pharmaceutical logistics also under the conditions of Slovak (Czech) pharmacy as early satisfaction of the requirements of the patient (client) in the pharmacy premises, which means that the appropriate pharmaceutical product or health care product and the appropriate information must be at the right time in the required amount and required quality in the right place. PMID- 16383188 TI - [Interconversion of stereochemically unstable chiral drugs: utilization of chromatographic techniques for the study of enantiomerization, calculation of thermodynamic parameters. Part I: theoretical aspects]. AB - The paper explains the theoretical aspects of the process of enantiomerization and describes its characteristic features (generation of a plateau). In addition, some complications are presented that are produced by enantiomerization either from analytical or pharmacological points of view. It also defines the way of how to calculate energy barriers of enantiomerization according to the methods used for the separation of racemic mixtures (stopped-flow or dynamic chromatography). Mathematic models useful in deconvolution of chromatograms are also described. With the use of dynamic methods it is not possible to quantitatively evaluate the obtained chtromatograms and calculate thermodynamic parameters without computer assisted deconvolution. PMID- 16383189 TI - [Composite tincture from the tops of the purple coneflower (Echinacea purpurea) and the fruits of the dog rose (Rosa canina): technology of production and evaluation of quality]. AB - This paper aimed to elaborate the technology of the production of a composite tincture with an immunostimulatory and adaptogenic effect, to select suitable methods of quality evaluation, and perform stability tests. The tops of the purple coneflower (Echinaceae herba) and fruits of the dog rose (Cynosbati fructus) in a ratio of 1:1 were employed to prepare the tincture. The tincture itself was prepared in a ratio of 1:5. The experiment revealed that ethanol 50% was the most suitable extracting agent. The optimal conditions for the obtaining of a quality tincture are formed when the drug is disintegrated into the required degree by grinding, percolation is used for extraction, and the rate of flowing of the extract from the percolator is 0.2 ml/min/100g of the drug. The determination of the dry residue, relative density, ethanol concentration and content of cichoric acid (spectrophotometrically), ascorbic acid (oxidimetrically), and tannins (manganometrically) served to evaluate the quality of prepared extracts. Stability tests (temperature 25+/-2 degrees C, relative humidity 60+/-5%, period of 2 years) have shown that the prepared tincture maintains its quality parameters for the given period. PMID- 16383190 TI - [Isotachophoretic determination of desloratadine in tablets and syrup]. AB - Desloratadine was determined in two dosage forms (tablets and syrup) with the use of the method of capillary isotachophoresis. Several electrolyte systems of varying composition and varying pH were tested. Two electrolyte systems were selected for the validation of the method and determination of desloratadine in real samples. In both electrolyte systems, the precision, correctness, linearity, robustness, and selectivity of the ITP method were evaluated. The pre-treatment of the sample prior to analysis consisted in dissolving and diluting the pertinent desloratadine-containing dosage form with demineralized water to the required concentration. This pre-treated sample was fed directly to the apparatus. PMID- 16383191 TI - [Analytical profile of 1-(4-fluorophenyl)-4-[3-(3-propoxyphenylcarbamoyloxy)-2 hydroxypropyl]piperazinium chloride (6f)]. AB - The paper presents the results of an analytical evaluation and a study of some physicochemical properties of a potential antiarrhythmic agent marked as 6f. The melting point, spectral properties, solubility and chromatographic behaviour of the drug on the thin layer were investigated. For the content determination, spectrophotometry in the ultraviolet region of the spectrum at the wavelength of the second absorption maximum of the substance and high-performance liquid chromatography were used. PMID- 16383192 TI - [Pharmaceutical literature in the library of the Kuks Hospice I. A contribution to the history of the hospital pharmacy of the Brethern of Mercy called "The Pomegranate" at Kuks]. AB - During the research of the history of the Baroque pharmacy of the Merciful Brethern at the Kuks Hospice (Eastern Bohemia), established in 1743, and its professional standard, pharmaceutical literature was found at the preserved hospice library. The present author first focused on the existence of basic, obligatory literature: dispensatories, tariffs of medicaments, and juridical literature (pharmaceutical laws). There survived a small number of these books because many books were destroyed during various wars. The following dispensatories (pharmacopoeias) were found: Dispensarorium pharmaceuticum Austriaco-Viennense 1729/1738, Dispensatorium Regium et Electorale Borusso Brandenburgicum 1744, Dispensatorium pharmaceuticum Pragensae renovatum 1750, Pharmacopoea Austriaco-provincialis 1774, 1775, 1790, 1794 (including a critical commentary by Z. G. Huszty 1797), The Military Dispensatory 1795, Pharmacopoea Austriaca 1812, 1814 and 1855. There are a few books of the tariffs of medicaments (only from 1795, 1816 and 1866) and juridical literature. These dispensatories (and their occurrence) will be useful for an analysis of laboratory preparation and supplying this pharmacy. A few illustrations of pharmaceutical literature, found bookplates, and a print of a painting of Count F. A. Sporck are attached. PMID- 16383193 TI - [Focus on the various techniques of anaesthesia in ophthalmologic surgery]. AB - Ophthalmologic surgery is not a vital one. It mostly concerns patients in extreme ages: children and old people. The risk of anaesthesia mainly depends on the health conditions of the patient. General anaesthesia through use of intravenous drugs alters the balance of the endocrine and sympathetic systems. Tracheal airway intubation constitutes an added constraint and stress especially for the vascular system. Retrobulbar anaesthesia is not devoid of risks: ptosis, diplopic, orbit haematoma, lesion of the optic nerve, eye perforation, vascular occlusion, intra arterial injection, neurologic and cardiovascular toxic effects of local anaesthetics, are all ever present risk Topical anaesthesia with its various variants (single topic or associated with intracameral injection, subconjunctival, circumferential perilimbal, subtenon) represents an interesting alternative for it is simple, less toxic and harless. In our experience, this method is indicated in surgery of eye's anterior segment. We find it safe, efficacisious and economical. Regional anaesthesia is preferred to general anaesthesia especially in the surgery of dacryocystitis and ptosis. General anaesthesia in stell indicated in case of children. PMID- 16383194 TI - [Prevalence and causal factors of anemia in children in Tunisia]. AB - Anemia continue to be prevalent among children under five years despite the improvement of socioeconomic and sanitary indicators. The purpose of the present cross-sectorial study is to assess the etiologic factors responsible for anemia. Knowledge of the relative importance of the different causes should form a basis for intervention strategies to prevent and control anemia. The survey covered 955 children under the age of five years, native of two regions with the highest prevalent of anemia, the Southwest and the District of Tunis. The results showed that 29% of children suffered from anemia. About 70% of them were iron deficient. The fractions of the deficiency in vitamin B12 and in folates were insignificant. Only 3% of children had chronic inflammation associated with (and possibly responsible for) their anemia A little fraction of anemia (approximately 5%) was due to thalassemia or drepanocytosis. Picawasan important causal factor of iron deficiency anemia. The parasites identified instool could not cause anemia. PMID- 16383195 TI - [Indications, complications and cost of internal jugular catheters in hemodialysed patients: study of 533 cases]. AB - INTRODUCTION: Central venous catheters for hemodialysis are very useful as blood accesses when arteriovenous fistulas (AVF) are not available. The aim of this study is to analyse the clinical impact of internal jugular catheters (IJC) in hemodialysed patients and to assess their economic consequences. PATIENTS AND METHODS: It is a prospective study realised from July 1998 to March 2002 including 533 hemodialysed patients without functional AVF: 280 males and 253 females aged between 17 and 87 years (mean age: 54.1 +/- 15 years). Single lumen polyurethane Vygon were used. All catheters were placed using Seldinger procedure and the posterior route of Jernigan which is more comfortable for patient. RESULTS: Indications of IJC placement were new hemodialysed patients without AVF in 73.5% cases and no functional AVF in 26.5% cases. During the period of the study. IJC was placed in 533 patients. We failed to place the IJC at the first attempt in 42 patients but we succeed in placing it on the controlateral side. During IJC placement, we observed 50 (9.4%) cases of accidental puncture of carotid artery. The median duration use of IJC was 41 days (extreme: 1 to 413 days). IJC were removed mainly because of the use of AVF in 469 (88%) cases and the catheter infection in 41 (77%) cases. Bacteriological analysis were made only for 25 patients: staphylococci were identified in 16 cases and catheter cultures were negative in the 9 other cases. All patients were treated with antibiotics. Outcome was favourable for 39 of them and we had 2 deaths by septicemia. The IJC cost was 44,287 Tunisian Dinars (DT) coresponding to 31.633 Euro including 10125 DT (7,232 Euro) for infectious treatment. CONCLUSION: The use of IJC is frequent in our center with a high rate of infectious complications which increased the hemodialysis cost. The realisation of AVF, in patients with chronic renal failure before the beginning of hemodialysis, is the best way to limit the use of catheters. PMID- 16383196 TI - [Latex allergy in a hospital environment: results of a study in Sousse (Tunisia)]. AB - Immediate allergy to natural latex has been on the icrease for the last decade in exposed persons and particularly the health personnel. Its prevalence is variable from one study to another. To evaluate the prevalence of later allergy in our nursing staff, we carried out an inguiry in two hospitals in Sousse. It's a prospective study involving 300 subjects and based on an anonymous questionnaire and a cutaneous prik test for latex allergy and atopy. The population was composed of 188 women an 112 men ith an average age of 35 years. There were 97 nurses of either sex, 65 doctors, 58 workers, 30 with technicians, 27 anesthetists, 17 midwives and 6 pharmacists. Sixty subjects (5.3%) showed a positive prick test ot latex: 11 women and 5 mon of different professional groups. The prevalence of atopy in this latex positive population (81%) was greater than in the latex negative population (25%). An associate alimentary allergy was identified in 10 patients allergic to latex. A study of specific immunoglobulins to latex carried out in 12 cases was positive in three patients. The provocation test for gloves was positive in 38% of cases. PMID- 16383197 TI - [Kaposi' s sarcoma. A series of 65 cases]. AB - Kaposi sarcoma (SK) is characterized by proliferation of vascular and spindle shaped cells with variable clinical presentations. The association with AIDS modifies the profile of this disease. The pathophysiology seems to be based on the major role HHV-8. The goal of this work was to specify the profile of this disease in our region through a retrospective study of 65 cases of SK over a 23 year period (1981-2003). In our series, the classic form prevailed 163 cases (96.9%)]. One patient had a secondary form due to steroid therapy and another patient had a KS associated with AIDS. A male prevalence was noticed (M/F: 2.8). Cutaneous lesions were dominated by papulo-nodular lesions (90.9%) of the lower limbs (92.3% of the cases). Mucous lesions were present in 15.4% of the cases. Therapeutic abstention was decided in 38.4%. Surgical excision was indicated in 35.4% of cases, radiotherapy in 4.6% of the cases and chemotherapy in 23% of the cases. The classic form of KS is the most frequent in our area; the other forms remaining rare. PMID- 16383198 TI - [Role of visceral obesity in metabolic disorders associated with hyperandrogenia in hirsute women]. AB - The aim of our study was to evaluate the relationship between the importance of visceral fat measured by tomodensitometry and the metabolic disturbances in hirsute women. It is a prospective study where 52 women were explored for hirsutism. They underwent a clinical and biological examination, as well as an abdominal tomodensitometry at the 4th and 5th lumbar spine level to measure the area of visceral fat (VF) and sub-cutaneous fat (ScF). In the 25 patients with VF predominance (VF/ScF > or = 0.2), age, body mass index (BMI), fasting insulin, (12h) and the prevalence of dyslipidaemia were significantly higher compared with women with ScF predominance (VF/ScF < 0.2). There was a positive correlation between VF and age. BMI, waist to hip ratio, 12h and triglyceridemia. In conclusion, hirsute women with visceral fat predominance are at higher risk to develop the insulin resistance syndrome. So, preventive measures against cardio vascular risk factors, like overweight, must be a part of the treatment. PMID- 16383199 TI - [Homocysteinaemia and degenerative complications in non insulin dependent diabetes mellitus]. AB - Type 2 diabetes mellitus (non insulin-dependent diabetes mellitus: NIDDM) is known to be associated with degenerative complications. Although, the pathophysiology of such complications is well known, the role of homocysteine (Hcy) is still discussed. The aim of the present study was to evaluate the relationship between the homocysteine levels and the NIDDM related complications in a group of NIDDM patients. Our study population consisted of 41 NIDDM patients including 13 subjects (G1) without complications (group controls), 17 patients (G2) with microangiopathy and 11 patients (G3) with coronary deficiency. Plasmatic homocysteine, glycemia, glycated haemoglobin (HbA1C) and lipidic parameters were essessed in all patients. Our results showed that mean levels of plasmatic homocysteine were within the normal range (10.4 +/- 3.3 micromol/l, 9.9 +/- 5.5 micromol/l and 14.8 +/- 10.4 micromol/l in G1, G2 and G3 respectively). Nevertheless, moderate hyperhomocysteinaemia was found in 36% in the coronary group (G3), 17.3% in patients with microangiopathy (G2) and 7.7% in controls. These preliminary results showed that cardiovascular complications in NIDDM patients may be related to high levels of homocysteine. PMID- 16383200 TI - [Preliminary results of surgical treatment of developmental dislocation of the hip in older children]. AB - Reduction of developmental dislocation of the hip is difficult to achieve in children after walking age and particularly in older children. In fact, at this age the important retraction of the muscles around the hip associated with a marked acetabular dysplasia and elongation of the joint capsule explain the difficulty and instability of reduction and the frequency of complications. In this study we reviewed retrospectively the clinical and radiological results of 26 developmental dislocations of the hip treated by open reduction, pelvic osteotomy and femoral shortening in 21 children aged more than 5 years. Age at surgery ranged from 5 to 11 years (mean 7.5 years) with a follow-up of 1 to 8 years (mean 2 years 7 months). According to the clinical classification of MC Kay, 17 hips had a good result while 9 hips had a fair or poor result. According to the Severin classification system 18 hips had an excellent and good radiological result. Ten hips developed an avascular necrosis of the femoral head following the reduction. PMID- 16383201 TI - [Identification of a new genetic entity in the form of an autosomal dominant axonal Charcot-Marie-Tooth disease associated with periodic paralyses and pyramidal syndrome]. AB - We report the clinical and genetic linkage analysis of eight affected patients suffering from axonal form of Charcot-Marie-Tooth disease (CMT2) with periodic paralyses and pyramidal feature. The inheritance is autosomal dominant. It was characterized by onset between the first and the second decade, distal weakness and atrophy of lower limbs. Four patients showed deep sensory loss; a periodic paralyses were also present in four patients. Genetic linkage excluded all known loci of CMT2 and other loci of clinical distinctive feature. Exclusion of these loci demonstred the distinctive phenotype of this family and assess the identification of a new genetic form. PMID- 16383202 TI - [Liver regeneration after major hepatectomy. Evaluation of dogs hepatectomy]. AB - OBJECTIVES: The aim of our study is to evaluate anatomic regeneration and metabolic derangement of the liver after major resection in dogs. METHODS: This is an experimental study on 9 dogs; we divided the dogs in two groups: the first group (5 dogs) underwent at one go major hepatectomy (90% of the liver). The second group (4 dogs) underwent successively a resection of 75% of the liver and a second resection of 90% of the restored liver six months later. All dogs underwent a metabolic and morphologic studies of the liver and of their kidney function. RESULTS: In the first group; all dogs which underwent 90% hepatic resection died 48 hours after the surgical resection of hepatic insufficiency. The ultra microscopic study showed the role of portal hypertension in hepatic degeneration on the first group. In the second group, the dogs survived the first resection, and our study shows a regeneration of the liver after resection and sub normal hepatic function. CONCLUSION: The liver is able to regenerate after minimally resection but major resection must be done by successively resection to avoid hepatic dysfunction, but the time between resection must be evaluate later. PMID- 16383203 TI - [Gastritis cystica polyposa: report of 7 cases and literature review]. AB - Gastritis cystica polyposa (GCP) is a rare inflammatory disease of the gastric remnant that usually develops after partial gastrectomy. It is defined by the presence of polyps on anastomotic gastric mucosa and at histopathological examination, by the presence of mucosal and submucosal cysts with foveolar hyperplasia. The disease is characterized by the frequency of gastro-intestinal bleeding and possibility of carcinomatous association. We report seven cases of GCP collected over 5 years (1994-1999). Diagnosis was made 9 to 45 years after partial gastrectomy and Finsterer. The patients were 52 to 72 years old. Revealing symptoms were cardiac failure, cardiac ischemia, melena, severe anemia and epigastric pain. For one patient, GCP was discovered casually. Endoscopic examination showed in all patients; the presence of several polyps sized between 3 to 15 mm on the perianastomotic gastric remnant. Histology examination of the polyps showed microscopic features of GCP in all cases. In one patient, there was a mild glandular atrophy with extensive intestinal metaplasia and mild dysplasia. Helicobacter pylori was present only in this case. PMID- 16383204 TI - [Rhinoscleroma: a report of 2 cases]. AB - Rhinoscleroma is a rare chronic granulomatous infection caused by Klebsiella rhinoscleromatis. Because of its mundane clinical presentation as chronic rhinits, it is frequently unrecognized. The authors report the cases of two female patients with pseudotumoral rhiniscleromas located in the septum and in the rhinopharynx respectively CT scan permitted a precise evaluation of the extent of the lesion. Among the various para-clinical examinations required, bacteriological studies are very important and should be performed comcomitantly with biopsy. PMID- 16383205 TI - [Reexpansion pulmonary edema a vacuo: a case report and literature review]. AB - Reexpansion pulmonary edema is a rare and potentially lethal complication with a 15 to 20% mortality. We report one case of pulmonary edema after evacuation of spontaneous right pneumothorax The pathophysiology of reexpansion pulmonary edema remains obscure; mechanical and inflammatory processes (production of Inerleukin 8--and Leukotrienne B4) seem incriminated. Duration and seventy of lung collapse and the rate of reexpansion appear to be the main causes justifying preventive measures. Curative treatment is based on oxygenation and lower aspiration pressure. PMID- 16383206 TI - [Gastrointestinal bleeding caused by rupture of an aneurysm of the hepatic artery]. AB - Gastrointestinal bleeding secondary to a rupture of an aneurysm of the hepatic artery is rare. We report a case of an 81-year-old man, who was admitted with hematemesis and melena. Gastroduodenoscopy revealed an abundant bleeding from the posterior wall of the duodenal bulb. CT-scan shoved an aneurysm arising from the hepatic artery. Ultrasound, computed tomography and angiography are the methods of choice for gastrointestinal bleeding diagnosis if endoscopy is inconclusive. PMID- 16383207 TI - [Iliac chondroma: a case report]. AB - Chondroma is a benign bone tumour that usually occurs in the in carpal and phalangeal bones. The iliac involvement is very rare and should be differentiated from chondrosarcoma. The authors report a case of iliac chondroma revealed by an acetabular lytic area in a 13-year-old girl. The diagnosis of chondroma was confirmed after surgical biopsy. PMID- 16383208 TI - Basal ganglia: functional perspectives and behavioral domains. PMID- 16383209 TI - Behavioral changes in patients with focal lesions of the basal ganglia. PMID- 16383210 TI - Depression in Parkinson's disease. AB - In contrast to depression, motor symptoms in PD have been extensively researched. The result is a detailed understanding of the pathophysiology of abnormal movements (162). Commitment to, and funding for, a well thought-out depression research plan, such as that presented by the working group at the National Institutes of Health (6), will bring similar theoretical understanding and patient relief for the most disabling of all PD symptoms (3-5)--depression. PMID- 16383211 TI - Anxiety disorders in Parkinson's disease. AB - Anxiety disorders frequently occur in association with PD and may be important causes of morbidity. Actual prevalence rates are uncertain, but estimates suggest that up to 40% of patients with PD experience substantial anxiety. This percentage is greater than expected, particularly for an elderly population. In addition, the age at onset of anxiety in PD (and particularly panic disorder) is later than would be expected from current information regarding the natural course of anxiety disorders. Virtually all of the types of anxiety disorders have been described in PD, but panic disorder, GAD, and social phobia appear to be the ones most commonly encountered. Although most patients with motor fluctuations experience greater anxiety during the "off" phase, this is not a universal phenomenon. Anxiety frequently develops before the motor features do, suggesting that anxiety may not represent psychological and social difficulties in adapting to the illness but rather may be linked to specific neurobiologic processes that occur in PD. Most evidence points to disturbances in central noradrenergic systems, but other neurotransmitters (e.g., serotonin, dopamine) may be involved as well. Studies suggest that right hemispheric disturbances may be particularly important for the genesis of anxiety, especially panic and OCD. Whether antiparkinsonian medications themselves contribute to anxiety needs clarification. Anxiety and depression frequently coexist in PD. It remains to be determined whether anxiety in patients with PD reflects one of the following pathologies: (a) an underlying depressive mood disorder, (b) a particular subtype of depression (atypical depression, anxious or agitated depression), or (c) an independent psychiatric disturbance. The relationship between anxiety and dementia in PD is not clear, but current evidence suggests that cognitive dysfunction is not related to the presence of anxiety symptoms in this disorder. The optimal pharmacologic treatment for anxiety in patients with PD has not been established, nor has the effect of PD surgery on anxiety symptoms. PMID- 16383212 TI - Disorders of motivation, sexual conduct, and sleep in Parkinson's disease. AB - Apathy and fatigue, sexual disturbances, mania, sleep disturbances, personality changes, pathologic gambling, and addiction to antiparkinson agents occur in patients with PD and may pose considerable stress on the patients themselves and their caregivers. With the exception of apathy and fatigue, little is known regarding the prevalence of these symptoms in patients with PD. The pathophysiologic mechanisms are unknown, although disturbances of the mesolimbic and mesocortical dopaminergic pathways are probably involved. Antiparkinsonian drugs or surgery seem to be the main etiologic factors for sexual disturbance, hypomania, addiction, and pathologic gambling, whereas hypodopaminergic states may contribute to symptoms such as apathy, fatigue, RBD, and possibly personality changes. Although some placebo-controlled trials have been published recently, no established treatments are currently available for these symptoms (85) (see Table 5-2), and thus future clinical trials are needed. PMID- 16383213 TI - Parkinson's disease as a model for psychosocial issues in chronic neurodegenerative disease. AB - Throughout the course of PD, opportunities abound for psychosocial interventions that can improve a patient's quality of life. The psychosocial challenges accompanying PD vary by stage of illness, though some challenges persist throughout the course of illness. As illustrated in the tables in this chapter, many of these interventions involve empathic listening and inquiry, education, challenging distorted or overly helpless thoughts and self-conceptions, and facilitating social contact and functioning. Others directly address the needs of caregivers who are confronted with the devastating effects of PD on their loved ones. Although many of the recommendations in this chapter are nonspecific and intuitive, their potential effect should not be underestimated. Although we have been unable to address all of the psychosocial issues confronted by patients facing PD, we hope that this chapter serves as a useful guide for providers looking for additional opportunities for psychosocial interventions designed to increase patient quality of life. These interventions can be delegated to other members of the team, and the assistance of psychiatrists or social workers may be warranted, but PD clinicians themselves can have the most dramatic effect on their patients because of their allocated authority and their salutary alliance with the patient. PMID- 16383214 TI - Early cognitive changes and nondementing behavioral abnormalities in Parkinson's disease. AB - Early cognitive changes in patients with PD are often subtle and influenced by factors that interact with the disease process, including age of disease onset, medication, and the specific constellation of motor symptoms. These factors notwithstanding, ample evidence exists that specific cognitive changes occur early in the course of PD. This evidence does not imply that cognitive deficits are pervasive during the early stages. To the contrary, they are usually subtle and often difficult to detect without formal neuropsychological testing. Executive-function deficits are the most frequently reported cognitive problems and, given that executive skills are an integral part of many tasks, it follows that subtle difficulties may be seen on a wide range of cognitive measures, particularly in working memory and visuospatial dysfunction, two areas that rely heavily on executive skills. Whereas apraxia and language processing deficits occur infrequently, subtle changes in olfaction and contrast sensitivity have also been repeatedly observed. Finally, depressive symptoms are also common in the early stages of the disease. The significance of the early behavioral changes and their prognostic implications are largely unknown. Prospective studies are needed to understand the longitudinal course of early cognitive changes to determine whether they remain as circumscribed impairments or represent a precursor to a more widespread dementia. PMID- 16383215 TI - Dementia in Parkinson's disease. AB - Dementia as a manifestation of idiopathic PD is an important feature of the disease in terms of QOL, prognosis, and clinical management. The pathophysiology of the disorder is likely a multifactorial process that encompasses derangement of multiple neuronal populations of both subcortical and cortical origin. This process results in a cognitive profile that largely reflects a dysexecutive syndrome. Currently, treatment of PDD is symptomatic. Further research is needed to elucidate the relationship of PDD to DLB and AD in hopes of developing and using appropriate therapeutic measures designed to address the underlying disease process. PMID- 16383216 TI - Behavioral changes as side effects of medication treatment for Parkinson's disease. PMID- 16383217 TI - Psychiatric symptoms following surgery for Parkinson's disease with an emphasis on subthalamic stimulation. AB - Bilateral subthalamic stimulation is a very effective neurosurgical treatment for advanced Parkinson's disease. Despite the range and frequency of psychiatric symptoms occurring in the postoperative state, most of these symptoms are transient and manageable. In clinical practice, preoperative psychiatric vulnerability, as with that of preoperative cognitive status, takes on an important role. Psychiatric assessment and active preoperative and postoperative intervention can potentially modify psychiatric outcomes. These psychiatric and psychological issues will take on greater importance, particularly with the rapid expansion of the number of neurosurgical sites and the need for adequate assessment and optimal management of patients. The paucity of the literature underscores the need for well-designed studies on psychiatric issues investigating both pathophysiology and clinical outcomes. PMID- 16383218 TI - Neurodegenerative disorders with diffuse cortical Lewy bodies. PMID- 16383219 TI - Cognitive and behavioral changes in the Parkinson-plus syndromes. PMID- 16383220 TI - Behavioral changes in frontotemporal dementia with Parkinsonism. PMID- 16383221 TI - Behavioral symptoms associated with Huntington's disease. AB - In addition to the abnormal movements most commonly associated with HD, cognitive impairment and psychiatric symptoms occur in almost all patients. These behavioral symptoms contribute to morbidity and augment what is often an overwhelming caregiver burden. Behavioral interventions may be useful in managing some HD patients with behavioral symptoms, especially irritability and apathy. Patients should be evaluated for underlying medical illness that might contribute to psychiatric symptoms, especially if onset of behavioral change is rapid. According to clinical observation, HD patients with psychiatric symptoms respond to standard pharmacotherapy. Controlled studies are needed to determine the true efficacy of these agents for use in treating people with HD. PMID- 16383222 TI - Psychosocial effects of predictive testing for Huntington's disease. PMID- 16383223 TI - Public health significance of tic disorders in children and adolescents. AB - In conclusion, data from community surveys suggest that tic disorders, including TS, exist on a spectrum from transient to persistent, multiple motor, and vocal tics that interfere with activities of daily living. The presence of isolated and transitory tics is common and appears to be of minimal consequence. On the other hand, persistent tics, even mild tics, appear to be associated with ADHD, disruptive behavior, and learning problems (though not necessarily formal learning disability). The presence of ADHD with tics increases the likelihood of disruptive behavior and learning problems, but learning problems can be observed in community samples of children with tic disorders, even in the absence of ADHD. To date, few studies have clearly defined the nature of the learning problems in children with tic disorders. Nonetheless, the data do suggest that having chronic tics is associated with impairment independent of ADHD. Community samples and recent investigations in clinical samples confirm that the presence of ADHD predicts greater disability than that associated with tic disorders alone. PMID- 16383224 TI - Obsessive-compulsive disorder in Tourette's syndrome. AB - A substantial body of scientific evidence suggests that obsessive-compulsive behavior occurs in a large percentage of patients with TS. Reliable estimates suggest that nearly 50% of patients with TS have some degree of obsessive compulsive features. Most patients with TS have only mild OCB and thus would not meet the DSM-IV diagnostic criteria for OCD. Therefore, OCB is perhaps a more appropriate characterization of this behavioral phenomenon that occurs in TS. OCB in TS appears similar to the spectrum of the tic disorder in terms of its onset, severity, and course. As with tics, OCB is typically mild and not always substantially disabling. Although clinical features between TS + OCB and primary OCD overlap considerably, patients with TS + OCB appear to experience different types of obsessive thoughts and compulsive rituals. Compelling genetic evidence suggests that OCB may be an alternative expression of the TS phenotype, which may selectively affect female gene carriers. Identifying the TS gene in the future will substantially broaden our knowledge of this intriguing neurobehavioral disorder. Finally, neurobiologic evidence points to similar anatomic and chemical substrates in the pathogenesis of TS and OCD, suggesting that these two disorders share a common pathophysiology. The clinical evaluation of patients with TS and their families should always include an assessment for OCB. Self-rated inventories of OCD such as the LOI, LOI-CV, and MOCI are useful screening scales. A more structured interview using the Y-BOCS (CY-BOCS) is useful for determining the degree and severity of OCB in TS as well as the response to therapy. Clinicians should keep in mind that OCB may be the most disabling feature of TS and may require treatment. Pharmacologic agents, such as SSRIs, and traditional behavioral therapy are proven effective treatments for OCB, which can substantially reduce the full effect of TS on patients and their families. PMID- 16383225 TI - Behavioral abnormalities in Wilson's disease. PMID- 16383226 TI - Hereditary ataxia and behavior. AB - Recognizing cognitive deficits and psychiatric disorders in patients with autosomal dominant ataxias is relatively new. At this time, the percentage of patients with these disorders who experience changes in cognition or psychiatric symptoms is unknown. Cognitive impairment, when seen, is often found on tests of executive function, probably reflecting disruption of afferent and efferent pathways of the prefrontal cortex and subcortical structures, including the cerebellum. Widespread global dysfunction does occur in some cases, especially later in the disease course. Psychiatric symptoms including depression, aggression, irritability, and psychosis have all been reported. As these behavioral changes receive further study, one hopes that guidelines for treating these symptoms will emerge. Clinicians should be mindful of the psychosocial effects that genetic testing for the hereditary ataxias may have, especially in cases of predictive testing for those who are asymptomatic but at risk because of family history. Guidelines established for genetic testing in HD may be helpful when approaching these cases. PMID- 16383227 TI - Behavioral symptoms associated with essential tremor. AB - As the complexity and heterogeneity of ET emerge, greater interest has been taken in the nonmotor manifestations of this disease. The fact that the motor manifestations reflect widespread anatomic pathology (e.g., cerebellum, basal ganglia) suggests that the nonmotor aspects might be a manifestation of an illness that affects multiple areas of the nervous system. Therefore, nonmotor aspects of the disease might reflect the presence of disease pathology in a particular area of the brain. Alternatively, nonmotor aspects could be the result of the tremor itself and the effects that it has on disability, mood, and personality. Finally, some of the nonmotor aspects could reflect the possibility that individuals with these disorders have a shared predisposition for ET and that a common mechanism underlies both. Further work is needed to define the extent of the nonmotor manifestations, their presence or absence in the predisease state, and their progression over time. These studies will further our insights into the mechanisms and anatomic pathology of ET and help us to develop a broader understanding of issues that are important in treating the individuals with this disease. PMID- 16383228 TI - Behavioral and psychiatric manifestations in dystonia. PMID- 16383229 TI - Autoaggressive immune-mediated movement disorders. AB - Poststreptococcal disorders exhibit a remarkable comorbidity of neurologic and psychiatric features. A similar combination of symptoms is also described in other conditions, such as connective tissue or paraneoplastic disorders, albeit less frequently. A better understanding of the underlying mechanisms associated with autoaggressive immune-mediated attack on the basal ganglia is required. This understanding will ideally aid clinicians in diagnosing these conditions and lead to appropriate clinical trials, for example, of chemoprophylaxis strategies to prevent recurrent streptococcal infection and of the use of immunosuppressive treatments. PMID- 16383230 TI - Psychopathological and cognitive correlates of tardive dyskinesia in patients treated with neuroleptics. AB - Even though new cases of TD are on the decline in North America and other western countries, TD remains a public health concern for patients with chronic schizophrenia, PAD, and for nonpsychiatric patients treated with dopamine receptor antagonists. The new generation of atypical antipsychotic medications is believed to pose less risk for TD. However, identifying the cognitive and disease related correlates of TD should equip clinicians with the necessary tools to reduce the prevalence of this iatrogenic movement disorder. No effective treatments for TD are available. This lack of effective therapy is problematic, especially in the few patients in whom the disorder causes functional impairment and other complications, and in whom it may be irreversible. PMID- 16383231 TI - Treatment issues in psychogenic-neuropsychiatric movement disorders. AB - Patients with PNMDs pose a fascinating challenge to clinicians at the neurology psychiatry interface. We have outlined a diagnostic and therapeutic approach to these complex disorders. Patients with PNMDs typically manifest abnormal movements and postures that do not fit expected patterns of movement disorder phenomenology. The first goal of neurologic consultation is to make an accurate diagnosis, with a view to either ruling out or defining the extent of any organic substrate. The next task is an effective referral to a psychiatrist capable of diagnosing relevant psychopathology and collaborating in a treatment plan. A supportive explanation of the diagnosis, one that is sensitive to the patient's intellectual capacity, conception of the illness, and self-esteem, appears to be a crucial determinant that influences the patient's capacity to be engaged effectively in treatment. The treatment for each patient with PNMD is individualized and may include psychotherapy, hypnosis, pharmacotherapy, physical therapy, and other approaches. To date, no treatment approach has been shown to be superior, and very few long-term outcome data are available. Although many factors influence response to treatment, those of particular importance are the effectiveness of communication among the neurologist, the psychiatrist, and the patient; the nature, severity, and chronicity of relevant psychopathology and environmental stressors; and the quality of external support and intrinsic resources available to the patient. PMID- 16383232 TI - [Effect of tyrosine kinase inhibitor Imatinib mesylate on proliferation, differentiation and apoptosis of Kasumi-1 leukemia cell line]. AB - OBJECTIVE: To explore the effect of Imatinib mesylate on proliferation, differentiation and apoptosis of leukemic Kasumi-1 cells bearing c-kit mutation. METHODS: Kasumi-1 cells were treated with Imatinib at different concentrations in culture. Cell proliferation was assayed by MTT assay, expressions of c-kit antigen, surface myeloid antigen and cell cycle by flow cytometry, cell apoptosis by annexin V staining and agarose gel electrophoresis. Western blot was used to analyze the level of c-kit protein tyrosine phosphorylation. RESULTS: Imatinib treatment caused a time- and dose-dependent inhibition of the cell proliferation, with a 72 h IC50 of 4.45 micromol/L. Imatinib treatment induced a decrease in the mean fluorescence value of c-kit antigen, a progressive decline in S-phase cell fraction and an increase in G0/G1 cells. Treatment with 5.00 micromol/L of imatinib for 72 h induced an increase in expression of myeloid surface protein CD11, CD13 and CD15, and for 24 h induced an increase in early apoptosis cells [from 9.04% to 86.84% (P < 0.05)]. The apoptosis ladder was observed on agarose gel electrophoresis on 5-day treatment. Tyrosine phosphorylation level of c-kit protein was decreased by Imatinib treatment. CONCLUSION: Tyrosine kinase inhibitor Imatinib mesylate treatment could inhibit proliferation of Kasumi-1 cells which bear a c-kit mutation, induce differentiation, apoptosis and G0/G1 cells accumulation. PMID- 16383233 TI - [Function analysis of the family-specific CTL induced by peptides derived from IgHV gene framework region of B-cell malignance]. AB - OBJECTIVE: To identify immune epitopes existed in the framework region (FR) of the IgHV protein of B-cell malignance, and explore the use of these FR-derived peptides to induce the family specific immune response in vitro, in order to explore the possibility of a new IgHV gene family-specific immunotherapy for B cell malignance. METHODS: Bioinformatics was used for predicting T cell epitopes in IgHV protein. Peptides of interest were synthesized in vitro. T2 cell binding assay was performed to determine the binding ability of the peptides to HLA A*0201 molecules. Peptide/HLA tetramer staining was used to detect the number of peptide-specific cytotoxic T lymphocytes (CTLs). Cytotoxicity assay was used to determine killing activity. RESULTS: Twelve peptides that were common to seven IgHV gene subfamilies were identified, and 10 (83%) of them were located in the FR of IgHV protein. The synthesized peptides up-regulated HLA*0201 molecules fluorescence intensity on cell surfaces of T2. By using an antigen-specific T cell expansion system in vitro, the peripheral blood mononuclear cells (PBMNC) from a healthy HLA-A0201 donor were stimulated weekly by autologous PBMNC loaded with the peptide as antigen presenting cells (APC), and the peptide-specific CTLs were demonstrated to be generated successfully in the healthy donors. The frequency of CD8 and peptide/HLA tetramer double positive cells in the gated lymphocyte population was 0.38% before stimulation and increased to 49.38 % after four times stimulation. Cytotoxicity assay indicated that these CTLs were capable of killing the HLA-A*0201, IgHV1 (+) lymphoma cells. Furthermore, the generated CTL could not kill the target cell loaded with the IgHV3 peptide, indicating that the cytotoxicity is family-specific. CONCLUSION: Peptides derived from the IgHV protein FR can successfully induce the generation of peptide-specific CTLs in vitro. These CTLs are capable of killing the lymphoma cell belonged to the same subfamily in a peptide-specific and MHC-restricted way. These findings could potentially form the basis of broadening application of immunoglobulin-directed immunotherapy in B-cell malignancies. PMID- 16383234 TI - [Effects of inhibiting SDF-1 expression by RNA interference on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells]. AB - OBJECTIVE: To observe the effects of inhibiting stromal cell derived factor-1 (SDF-1) expression by RNA interference (RNAi) on adhesion and drug sensitivity of Jurkat cells co-cultured with bone marrow stromal cells. METHODS: SDF-1 specific short hairpin RNA (shRNA) expressing plasmid was transferred into cultured human acute leukemic bone marrow stromal cells, positive clones were isolated by screening G418 resistance (Group A) , SDF-1 protein level in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The adhesion rates to bone marrow stromal cells layer and the drug sensitivity to doxorubicin of co cultured Jurkat cells were detected by cell counting and MTT assay, respectively. The un-transfected bone marrow stromal cells of acute leukemia patient (Group B) or normal subject (Group C) were taken as control. RESULTS: The level of secreted SDF-1 protein (pg/10(5) cells/week) in the supernatants of Group A, B and C were 1920 +/- 205, 12,370 +/- 1355 and 6620 +/- 770, respectively. Of co-cultured Jurkat cells in Group A, B and C, the adhesion rates after 24 h co-culturing were (28.8 +/- 2.6)%, (57.4 +/- 3.8)% and (45.2 +/- 4.0)%, respectively, and the IC50 values of doxorubicin were 585, 6162 and 1758 nmol/L, respectively. CONCLUSION: Down-regulating SDF-1 expression of bone marrow stromal cells by RNAi reduces adhesion rates and enhances drug sensitivity to doxorubicin of their co-cultured Jurkat cells. PMID- 16383235 TI - [Induction of efficient T-cell immunity against autologous leukemia cells by dendritic cells pulsed with the leukemia cell total RNA]. AB - OBJECTIVE: To assess the feasibility and efficiency of eliciting leukemia specific T cell responses in acute myeloid leukemia patients in complete remission (AML-CR) in vitro by dendritic cells (DC) pulsed with the leukemia cells total RNA. METHODS: The immature DCs were generated from the adherent bone marrow mononuclear cell in vitro in the presence of combined cytokines (GM-CSF 100 ng/ml, IL-4 500 U/ml), and pulsed with total RNA isolated from autologous leukemic cells by cationic lipid 1,2-dioleoyloxy-3-trimethyl ammonium propane (DOTAP) at day 5 of culture. Then the cells were incubated for another 24 h in a medium containing 10 ng/ml of TNF-alpha for maturation of DC. After the total 7 days culture, the cells were harvested as the mRNA-DC and the expression of mature DC markers were determined by FACS. The proliferative capacity of T cell activated by mRNA-DC was determined by MTT assay. Meanwhile, the mRNA-DC was co cultured with T lymphocytes at a ratio of 1:3 for 7 days. The activated T lymphocytes were harvested, the secretion of IFN-gamma was determined by ELISPOT assay, and the cytotoxicity was analyzed in vitro by LDH release assay. RESULTS: After culture, the BMMNC from 14 AML-CR patients developed morphologic and phenotypic characteristics of mature DC. At a stimulator/reactor ratio of 1:16, auto-T lymphocytes primed with mRNA-DC exhibited significant proliferative activity compared with T lymphocyte primed with non-pulsed DC [(36.84 +/- 5.68)% vs (12.20 +/- 3.16)%, (P < 0.05)]. An expansion of mRNA reacted T cell secreting IFN-gamma could be observed on ELISPOT assay. At an effector/target ratio of 20:1, the auto-T lymphocytes primed with mRNA-DC exhibited significant killing activity to auto-AML cells (45.46 +/- 6.34 )% as compared with that stimulated by IL-2 alone (13.26 +/- 2.28)% or primed by non-pulsed DC (12.32 +/- 1.32)% (P < 0.05). CONCLUSION: Immunization with DC-leukemia cell RNA vaccines may be a simple, rapid and potent approach to elicitation of T cell-mediated anti-leukemia immunity. PMID- 16383236 TI - [Recipient lymphopenia state enhances the expansion and anti-leukemia effect of leukemia specific cytotoxic T lymphocytes]. AB - OBJECTIVE: To determine the role of recipient lymphopenia state in the expansion and function of leukemia specific cytotoxic T lymphocytes (CTLs). METHODS: C57BL/6 mice were induced to lymphopenia with 6 Gy total body irradiation. Spleen T cells or leukemia specific T cells from EGFP+ transgenic C57BL/6-EGFP mice were adoptively transferred by intravenous injection. The mice were challenged subcutaneously with 1 x 10(6) FBL3 leukemic cells at day 2 after irradiation. The peripheral WBC count, percentage of EGFP+ cells, subsets of T cells and tumor sizes were monitored. RESULT: Both of the spleen T cell and leukemia specific CTL proliferated efficiently with the percentage of EGFP+ cells of 28. 81% and 42.24%, respectively, after infused into lymphopenic recipients. However, spleen T cells had no anti-leukemia effect regardless of its expansion. In contrast, leukemia specific CTLs showed a more rapid and extensive expansion under the condition of lymphopenia and a enhanced anti-leukemia immunity. CONCLUSION: Transfusion of leukemia specific CTLs under lymphopenia state could be a feasible strategy to expand leukemia specific CTLs and generate favorable anti-leukemia effect in vivo. PMID- 16383237 TI - [Comparative study on the treatment effects with rhIL-11 and rhG-CSF in combination or alone on immune function]. AB - OBJECTIVE: To investigate the effect of in vivo administration of rhG-CSF and/or rhIL-11 on mice immune system function. METHODS: T cell subgroups, suppressor T cells (CD8+ CD28-, CD4+ CD25+, CD3+ CD4- CD8- T cells), expression of CD28 on T cells, and spleen T cells intracellular IL4/IFN-gamma secretion were determined by multicolor flow cytometry. MTT was used to determine the T cell proliferation capacity and mixed lymphocyte reactions. RESULTS: In vivo administration of cytokines decreased the percentage of lymphocytes (P < 0.05), rhIL-11 and rhG-CSF in combination significantly decreased the CD4+/CD8+ ratio and increased the percentage of CD8+ CD28- suppressor T cells compared to either cytokine alone (P < 0.01). There was no difference in the percentage of CD3+ CD4- CD8- and CD4+ CD25+ suppressor T cells between either of the cytokines. Furthermore, cytokines treatments significantly decreased the capacities of splenic T cells proliferation and the response to alloantigens compared with the PBS treatment (P < 0.05), the combination group being more significantly decreased (P < 0.01). And cytokines treatment significantly decreased the production of IFN-gamma and increased the production of IL-4 compared with the PBS treatment(P < 0.05). The ratio of IFN-gamma/IL-4 were significantly decreased after the combination compared with either of them alone. CONCLUSION: The combination of rhIL-11 and rhG-CSF is potentially synergistic in the induction of immune tolerance by their effects on the proliferation capacity and function of T lymphocytes. PMID- 16383238 TI - [Study on the burden of abnormal hematopoietic clone of the patients with myelodysplastic syndromes and its implications]. AB - OBJECTIVE: To investigate the abnormal hematopoietic clone burden of the patients with myelodysplastic syndromes (MDS) and its clinical implication. METHODS: The ratio of the metaphase with abnormal karyotypes to the total was regarded as the index of MDS clonal burden. Thirteen parameters were assayed and the correlations between these parameters and MDS clone burden were analysed. RESULTS: The clonal burden of MDS patients was (67.4 +/- 36.2)%. It correlated positively with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = 0.445, P < 0.05). The number of blasts, hemoglobin and erythrocytes in high clonal burden (>50%) and low clonal burden (< or = 50%) groups were significantly different (P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (274.18 +/-71.85) x 10(6)/L and (454.82 +/- 205.88) x 10(6)/L (P < 0.05) respectively. CD8+ T lymphocytes between MDS patients and normal controls had no difference. The serum level of IL-2 of MDS patients and normal control groups were (6.29 +/- 3.58) g/L and (3.11 +/- 1.40) microg/L (P < 0.05) respectively; but no difference in the serum level of TNF between MDS and control groups. The ratio of CD4+ to CD8+ in high clonal burden patients was 1.90 + 0.52, and in low clonal burden patients was 0.97 +/- 0.44 (P < 0.05). CONCLUSION: The clonal burden and deficient T cell immunity are the indicators for predicting MDS patients clinical progression. PMID- 16383239 TI - [Study on clinical and biological characteristics of childhood acute leukemia with MLL gene rearrangements]. AB - OBJECTIVE: To study the clinical and laboratory features of childhood acute leukemia (AL) with MLL gene rearrangements. METHODS: Sixteen of 298 cases of childhood AL with MLL rearrangements were studied by using MLL dual-color FISH, multiplex RT-PCR with 13 pairs of primers in combination with R banding karyotype analysis and cell immunophenotyping by flow cytometry. RESULTS: Sixteen cases of childhood AL with MLL rearrangements accounted for 5.4% of 298 AL patients, and 56.3% of infant ALs. Among 106 cases analyzed by multiplex RT-PCR, MLL gene rearrangements were found in 11 cases, including MLL/AF4 fusion gene in 2, MLL/AF6 fusion gene in 1, MLL/AF6 and MLL/ELL combined with MLL/ AFX or HOX11 in one case each, MLL/AF9 in 2, MLL/AF10 in 1, MLL/ELL in 2. MLL partial tandem duplication in 1 and activated HOX11 in 1. In 27 cases assayed by FISH, 9 cases (36.0%) were demonstrated MLL gene rearrangements. In 16 patients with MLL gene rearrangements, 14 (87.5%) exhibited clonal chromosome abnormalities involved chromosome 11 in 11 cases: being t(4;11) in 2, t(6;11), t(8;11), t(7;8;11), t(9;11) in each trisomy 11 in 2 and 11q--in 3 cases. Among these 16 patients, 11 were B-ALL, and 5 AML-M5, 3 of the latter were CD7+ and CD2+. Of these 16 patients, 8 received chemotherapy and 7 of them achieved complete remission, while the other 8 patients gave up treatment. CONCLUSION: Multiplex RT-PCR combined with FISH provided a more accurate and sensitive method for detection of MLL gene rearrangements. Finding out MLL gene rearrangement is of most importance in guiding therapy and predicting prognosis in childhood AL. PMID- 16383240 TI - [Studies of treatment strategy and prognosis on acute myeloid leukemia with chromosome 8 and 21 translocation]. AB - OBJECTIVE: To investigate the relationship between the biological features and the treatment efficacy and prognosis in acute myeloid leukemia subtype M2 (AML M2) patients with chromosome 8 and 21 translocation. METHODS: By using Cox regression model and Kaplan-Meier analyses, prognostic factors in 54 cases of de novo adult AML with t(8;21) in our institute from 1990 to 2003 were retrospectively analyzed. RESULT: The complete remission (CR) rates were 81.9% for all M2 patients, 82.4% for patients with normal karyotype, 88.5% for patients with t(8;21) [P > 0.05 for normal karyotype vs t(8;21)], 100.0% for 28 patients with t(8;21) alone and 75.0% for 24 patients with additional chromosome abnormalities (P < 0.01). The actuarial 3 year overall survival(OS) was 26% for M2 patients with normal karyotype, 25% for patients with t(8;21) [P > 0.05 for normal karyotype vs t(8;21)], in whole t(8;21) group, 46.4% for patients with t(8;21) alone and 0% for patients with additional chromosome abnormalities (P < 0.01). Multivariate analysis of prognostic factors showed that chromosome abnormalities besides t(8;21) was the only factor affecting CR, disease-free survival (DFS) and OS. DFS of allogeneic hematopoietic stem cell transplantation (HSCT) and intermediate-dose cytarabine/high dose cytarabine (IDAC) groups were better than the group received routine dose cytarabine as postremission therapy (P < 0.01). CONCLUSION: AML with t(8;21) is not a single defined AML subset, and patients with additional chromosome abnormalities have a worse prognosis. HSCT and IDAC could improve the outcome. HSCT is the best choice for patients with high risks, especially with additional chromosome abnormalities. PMID- 16383241 TI - [Clinical and experimental study of 7 cases of acute lymphoblastic leukemia with dic(7;9) (pll;pll)]. AB - OBJECTIVE: To investigate the laboratory and clinical features of 7 cases of acute lymphoblastic leukemia (ALL) with dic(7;9) (pll;pll). METHODS: Cytogenetic examination of bone marrow cells was performed by direct method or short-term culture method. R banding technique was used for karyotype analysis. bcr/abl fusion gene was detected by interphase FISH using dual-color bcr/abl probe in 6 cases. FISH using chromosome 7-specific alpha-satellite DNA probe and chromosome 9-specific alpha-satellite DNA probe and chromosome painting using whole chromosome 7 and 9 paints probes were performed respectively. RESULTS: Seven (0.88%) of 800 ALL patients were found to have dic(7;9) abnormality. Among them, dic(7;9) was the sole abnormality in 2 cases, t(9;22), other additional aberrations besides dic(7;9) in 4 cases and dic (7;9) with other abnormalities but no t(9;22) in one case. Hyperleukocytosis (> 100 x 10(9)/L) was found in 4 cases with dic(7;9) and t(9;22), and patients without t(9;22) had WBC < 100 x 10(9)/L. Enlargement of liver, spleen and/or lymph nodes were found in 6 cases. Immunophenotyping showed that 5/6 cases of dic (7;9) ALL were of B lineage. Dual color FISH detected bcr/abl rearrangement in 3/6 cases and confirmed that the centromere of the derivative chromosome was originated from both chromosomes 7 and 9. A reciprocal translocation between chromosomes 7 and 9 was proved by chromosome painting. CONCLUSION: dic(7;9) was a rare, but recurrent chromosome abnormality in ALL and had some clinical and laboratory features. PMID- 16383242 TI - [Preliminary study of VEGF and its receptor expression on childhood acute lymphoblastic leukemia and its relativity to clinical manifestations]. AB - OBJECTIVE: To investigate the relationship between differential expression of VEGF and its receptors and clinical characteristics of childhood acute lymphoblastic leukemia (ALL). METHODS: Expressions of VEGF and its receptors (Flt 1, KDR) were assayed by ELISA and RT-PCR in healthy donors(20 cases), ALL patients in remission (20 cases), with low risk (29 cases) and with high risk (10 cases). The clinical data of all the patients and volunteers enrolled in this study were collected and analyzed according to the expression of VEGF and its receptors. RESULTS: The expressions of VEGF were (574.37 +/- 208.45) ng/L, (387.93 +/- 175.86) ng/L, (135.80 +/- 111.28) ng/L and (91.16 +/- 41.34) ng/L in patients with high risk, standard risk, in remission and healthy donors, respectively. Expression levels of VEGF receptors were downwards with risk grades. The clinical manifestations were also in accord with the expression levels of VEGF and its receptors. CONCLUSION: ALL patients with highly expressed VEGF and its receptors are usually with higher tumor burden, and refractory treatment. Detection of VEGF and its receptors might be one of prognostic marker for ALL treatment. PMID- 16383243 TI - [Expression of PTEN mRNA in acute leukemia and its clinical significance]. AB - OBJECTIVE: To explore PTEN gene expression and its clinical significance in acute leukemia. METHODS: The expression levels of PTEN mRNA in 5 leukemia cell lines, 87 patients with acute leukemias (AL), including 59 acute myeloid leukemia (AML), 26 acute lymphoblastic leukemia (ALL), and 2 acute hybrid leukemia, 21 AL in complete remission (AL-CR), 31 chronic myelogenous leukemia (CML) and 14 normal controls were assayed by RT-PCR. RESULTS: PTEN mRNA was detected in K562 cell line, but not in Kasumi-1, HL-60, U937, Nalm-6 cell lines. The expression ratio of PTEN mRNA between CML (61.29%) and normal control (78.57%) had no statistical difference (P > 0.05). The expression ratios of PTEN mRNA in AL (18.39%) and AL CR (42.86%) were significantly lower than that in normal control (P < 0.01 and P < 0.05, respectively), AL also has a lower expression ratio than that of AL-CR (P < 0.05). The decreased level of PTEN mRNA had a positive correlation with poor prognostic factors (high white blood cell count of > or = 20 x 10(9)/L and chromosome abnormality). CONCLUSION: There is down-regulated expression of PTEN gene in AL. PTEN gene may play a role in leukemogenesis. PMID- 16383244 TI - Comparison of endoscopic third ventriculostomy alone and combined with choroid plexus cauterization in infants younger than 1 year of age: a prospective study in 550 African children. AB - OBJECT: The aim of this prospective study was to determine whether, and in which patients, the outcome for bilateral choroid plexus cauterization (CPC) in combination with endoscopic third ventriculostomy (ETV) was superior to ETV alone. METHODS: A total of 710 children underwent ventriculoscopy as candidates for ETV as the primary treatment for hydrocephalus. The ETV was accomplished in 550 children: 266 underwent a combined ETV-CPC procedure and 284 underwent ETV alone. The mean and median ages were 14 and 5 months, respectively, and 443 patients (81%) were younger than 1 year of age. The hydrocephalus was postinfectious (PIH) in 320 patients (58%), nonpostinfectious (NPIH) in 152 (28%), posthemorrhagic in five (1%), and associated with myelomeningocele in 73 (13%). The mean follow up was 19 months for ETV and 9.2 months for ETV-CPC. Overall, the success rate of ETV-CPC (66%) was superior to that of ETV alone (47%) among infants younger than 1 year of age (p < 0.0001). The ETV-CPC combined procedure was superior in patients with a myelomeningocele (76% compared with 35% success, p = 0.0045) and those with NPIH (70% compared with 38% success, p = 0.0025). Although the difference was not significant for PIH (62% compared with 52% success, p = 0.1607), a benefit was not ruled out (power = 0.3). For patients at least 1 year of age, there was no difference between the two procedures (80% success for each, p = 1.0000). The overall surgical mortality rate was 1.3%, and the infection rate was less than 1%. CONCLUSIONS: The ETV-CPC was more successful than ETV alone in infants younger than 1 year of age. In developing countries in which a dependence on shunts is dangerous, ETV-CPC may be the best option for treating hydrocephalus in infants, particularly for those with NPIH and myelomeningocele. PMID- 16383245 TI - Long-term results of gamma knife surgery for the treatment of craniopharyngioma in 98 consecutive cases. AB - OBJECT: The authors analyzed the long-term outcomes of gamma knife surgery (GKS) for residual or recurrent craniopharyngiomas after microsurgery and the effects of dose reduction. METHODS: A total of 107 patients with craniopharyngiomas were treated with GKS at Komaki City Hospital during the past 12 years, and 98 patients were followed up for 6 to 148 months (mean 65.5 months). The mean tumor diameter and volume were 18.8 mm and 3.5 ml, respectively. These tumors were treated with a maximal dose of 21.8 Gy and a tumor margin dose of 11.5 Gy by using a mean of 4.5 isocenters. Final overall response rates were as follows: complete response 19.4%, partial response 67.4%, tumor control 79.6%, and tumor progression 20.4%. Reducing the tumor margin dose resulted in decreased therapeutic response and increased tumor progression, although the rate of visual and pituitary function loss also decreased. Among the factors examined, age (for adults) and the nature of the tumor (cystic or mixed) were statistically significant favorable and unfavorable prognostic factors, respectively. The actuarial 5- and 10-year survival rates were 94.1 and 91%, respectively. The progression-free survival rates were 60.8 and 53.8%, respectively. Patient outcomes were reportedly excellent in 45 cases, good in 23, fair in four, and poor in three; 16 patients died. Deterioration both in vision and endocrinological functions were documented as side effects in six patients (6.1%). CONCLUSIONS: Stereotactic GKS is safe and effective, in the long term, as an adjuvant or boost therapy for residual or recurrent craniopharyngiomas after surgical removal and has minimal side effects. New treatment strategies must be devised to manage these tumors. PMID- 16383246 TI - Central nervous system meningiomas in the first two decades of life: a clinicopathological analysis of 87 patients. AB - OBJECT: The occurrence of meningiomas in children younger than 20 years of age is rare, accounting for less than 3% of all childhood tumors of the central nervous system. The authors of this study sought to add to the limited available information regarding clinicopathological factors that influence outcome, disease progression, and survival in children with meningiomas. METHODS: Eighty-seven cases of childhood meningiomas were identified and classified according to World Health Organization (WHO) 2000 criteria. In addition to the WHO classification, the following potential prognostic factors were analyzed: age, sex, extent of resection, history of radiotherapy, diagnosis of neurofibromatosis Type 2 (NF2) or other inherited syndromes, and the presence of a comorbidity. There was a sex predilection for male patients (35 females and 52 males). Patient age ranged from 5 months to 20 years (mean 14 years). The most common clinical presentations were seizures (33%), headaches (13%), ataxia (10%), and hemiparesis (10%). Nine patients had NF2 and two had Gorlin syndrome. Seven patients had undergone radiotherapy for a prior neoplasm. Tumor location was supratentorial in 64% of the patients, infratentorial in 16%, intraventricular in 12%, and spinal in 8%. Fifty-three patients (62%) underwent gross-total resection and 28 (33%) underwent subtotal resection. Histopathological analysis revealed 62 (71%) WHO Grade I, 21 (24%) Grade II, and four (5%) Grade III meningiomas. One patient received adjuvant chemotherapy and four received radiotherapy. CONCLUSIONS: Using survival data from this unique patient cohort, the authors found that recurrence-free survival time was significantly related to WHO grade (p = 0.002), but overall survival time was not significantly linked to any of the potential prognostic factors considered in this study (p = 0.06). PMID- 16383247 TI - Quantitative pupillometry: normative data in healthy pediatric volunteers. AB - OBJECT: The authors describe the prospective use of a new hand-held point-and shoot pupillometer (NeurOptics) to assess pupil function quantitatively. METHODS: Repetitive measurements were made in 90 pediatric participants ranging in age from 1 to 18 years, providing a total of 100 measurements under ambient light conditions. The participants consisted of 45 patients without known intracranial or ophthalmological pathological conditions as well as 45 volunteers in the outpatient setting. Quantitative pupil measurements were reliably replicated in the study participants. The mean resting pupil aperture was 4.11 mm and the minimal diameter after stimulation was 2.65 mm, resulting in a 36% change in pupil size. The mean constriction velocity was 2.34 mm/second, with a mean dilation velocity of 2.2 mm/second. CONCLUSION: Pupil symmetry was impressive in the entire cohort. PMID- 16383248 TI - Early programmable valve malfunctions in pediatric hydrocephalus. AB - OBJECT: The use of adjustable differential pressure valves has been recommended to improve ventriculoperitoneal (VP) shunt performance in selected patients; however, published data are scarce regarding their clinical reliability. Recently, the identification of a number of malfunctioning programmable valves during shunt revision surgery in children prompted a retrospective review of valve performance in this patient cohort. METHODS: The authors performed a retrospective chart analysis of 100 patients with programmable valve shunts and 89 patients with nonprogrammable valve shunts implanted at the St. Louis Children's Hospital between April 2002 and June 2004. They noted the cause of hydrocephalus, the type of shunt malfunction, and cerebrospinal fluid (CSF) protein levels. Regular clinical follow up ranged from 1 to 26 months, with a mean follow-up time of 9.75 months for patients with programmable valves and 10.4 months for patients with nonprogrammable valves. Patient ages ranged from 2 weeks to 18 years. One hundred patients had 117 programmable valves implanted, and 35 of these patients (35%) underwent shunt revision because of malfunction. The programmable valve itself malfunctioned in nine patients who had undergone shunt revision (11.1%/year of follow up). The nonprogrammable valve group had no valve malfunctions. The overall VP shunt revision rate in the nonprogrammable valve group was 20.2%. No significant differences were identified when CSF protein levels and specific malfunction types were compared within the programmable valve and nonprogrammable valve groups. CONCLUSIONS: In this study the authors demonstrated an annualized intrinsic programmable valve malfunction rate of 11.1%, whereas during the same period no intrinsic valve malfunctions were noted with nonprogrammable valve systems for similar causes of hydrocephalus. The CSF protein levels did not correlate with observed valve malfunction rates. Further evaluation in a prospective, randomized fashion will elucidate specific indications for programmable valve systems and better determine the reliability of these valves in the pediatric population. PMID- 16383249 TI - Age-related variations in peak cerebrospinal fluid velocities in the foramen magnum. AB - OBJECT: Measurements of cerebrospinal fluid (CSF) velocities in the foramen magnum are used to determine the functional significance of Chiari malformation Type I (CM-I). Significantly higher peak velocities are found in adult patients with CM-I than in adult control participants. In addition, it appears that higher CSF velocities are found in pediatric patients with CM-I than in adult patients. Variations in CSF velocities across age groups in healthy individuals, however, have not been systematically studied. METHODS: Phase-contrast magnetic resonance imaging was performed in 10 pediatric patients after induction of anesthesia to evaluate conditions thought not to affect CSF flow in the foramen magnum. The peak systolic and diastolic velocities were plotted with respect to age and compared with velocities obtained in a group of 10 healthy adult volunteers. Differences between the adult and pediatric groups were tested for significance by using the Student t-test. Peak velocities ranged from 1.9 to 19.9 cm/second in the pediatric group and from 1.2 to 4.5 cm/second in the adults. A trend line fitted to the data showed a decrease in velocity with age in the first two decades of life, and little change thereafter. Differences in the pediatric and adult groups were significant at a level of 0.05. CONCLUSIONS: Peak CSF velocities vary significantly with age. To determine the normalcy of a CSF flow measurement, it must be compared with age-appropriate normative data. PMID- 16383250 TI - Neurenteric cysts in children: 16 consecutive cases and review of the literature. AB - OBJECT: The authors of this retrospective review and analysis of the literature cover an institutional series of neurenteric cysts of the central nervous system in children treated in the magnetic resonance imaging era during a 14-year period. METHODS: Sixteen patients 20 days to 14 years of age are described. The most frequent signs and symptoms at presentation were acute spinal cord compression (11 patients), paresis of a cranial nerve (two patients), meningitis or infection (two patients), and intracranial hypertension (one patient). The locations of the cysts were as follows: in the spinal canal in 12 patients (75%), the clivus in two (12.5%), the cavernous sinus in one (6%), and the craniocervical junction in one (6%). The most common location was the ventral aspect of the spinal canal (seven patients). Associated spinal deformities were found in five patients. All patients underwent surgery, with a posterior approach used in all of the spinal cases. Total resection was achieved in 12 of the 16 cases and partial resection in four. Of the four patients who underwent subtotal resection, the cyst recurred in three, requiring further surgery. CONCLUSIONS: Neurenteric cysts are uncommon congenital anomalies that can present acutely in the pediatric population. Total removal is usually possible and is associated with a good prognosis. PMID- 16383251 TI - Immature teratomas of the central nervous system: is adjuvant therapy mandatory? AB - OBJECT: Immature teratomas of the central nervous system (CNS) are rare neoplasms. Although adjuvant therapy is generally recommended after resection, the exact role of each therapeutic modality is not yet established. The purpose of this study was to analyze the clinicopathological correlation and the role of resection to define the optimal treatment modalities for immature teratomas of the CNS. METHODS: Between 1987 and 2002, eight patients underwent radical surgery for a lesion diagnosed as a CNS immature teratoma at the authors' institution. The clinical courses of these patients and the pathological features of their tumors were retrospectively reviewed. Gross-total resection (GTR) was achieved in six patients at the initial operation. The mean follow-up period was 75 months. Two patients received postoperative adjuvant therapies and two patients did not, against medical advice. None of the four patients experienced recurrence after long-term follow up. Another four patients, all of whom underwent GTR of the tumor, did not receive adjuvant therapy as part of a prospective treatment scheme. One of them exhibited early recurrence and metastasis. The tumor had pathological features denoting a high-grade (Norris Grade III) lesion and neurocytomatous differentiation. CONCLUSIONS: Aggressive resection seems to be of utmost importance in the treatment of immature teratomas of the CNS. Adjuvant chemotherapy and radiotherapy can be deferred if GTR is achieved in low-grade, immature teratomas, but adjuvant therapies may be warranted for high-grade ones. PMID- 16383252 TI - Split cord malformations: a clinical study of 254 patients and a proposal for a new clinical-imaging classification. AB - OBJECT: Split cord malformations (SCMs) are rare anomalies of the spinal cord. A total of 254 cases of SCM were treated surgically at the authors' center during a period of 16 years. METHODS: Patients' demographic profiles, imaging studies, operative details, complications, and surgical outcomes were evaluated retrospectively. A new classification based on intraoperative findings is proposed. The mean age of the patients was 7.3 years (female/male 1.5:1). Type I SCM was seen in 156 patients (61.4%) and 98 patients (38.6%) had Type II SCM. Skin stigmata were present in 153 cases (60%); hypertrichosis, being the most common, was seen in 82 cases (32.3%). Asymmetrical lower-limb weakness and sphincter disturbances were present in 173 (68.1%) and 73 (33%) cases, respectively. Of the symptomatic cases, 39% (68 of 173) showed improvement in motor power, 57.9% (33 of 57) experienced sensory improvement, and 27.3% (20 of 73) regained continence. None of the 38 patients in the asymptomatic group had postoperative neurological deterioration. The neurological status was unchanged in 63% of the cases. A new subclassification of Type I SCM is proposed, based on intraoperative location of a bone spur causing the split, which may have a bearing on surgical dissection and outcome. Based on the authors' experience with 25 cases of Type I SCM, they have classified the disorder into four subtypes: Type Ia, bone spur located in the center with duplicated cord above and below the spur (12 cases); Type Ib, bone spur at the superior pole with no space above it (four cases); Type Ic, bone spur at the lower pole with large duplicated cord above (three cases); and Type Id, bone spur straddling the bifurcation with no space above or below the spur (six cases). The risk of injury to the hemicords is highest in the Id subtype (four of six patients in this group deteriorated neurologically in the present series, whereas none with subtypes Ia-c worsened). CONCLUSIONS: The authors present the largest series on SCMs so far reported in the world literature The risk of neurological deficits developing increases with age; hence, all patients with SCM should be surgically treated prophylactically even if they are asymptomatic. This new classification is easy to use and remember and takes into account the use of intraoperative findings that may have a bearing on surgical outcome. PMID- 16383253 TI - Usefulness of constructive interference in steady-state magnetic resonance imaging in the presurgical examination for lumbosacral lipoma. AB - OBJECT: The aim of this study was to evaluate three-dimensional Fourier transformation constructive interference in steady-state (CISS) imaging as a preoperative investigation of anatomical structures around the lumbosacral lipoma for surgical untethering of the spinal cord and debulking of the lipoma. METHODS: The CISS imaging was used to visualize neural structures around the lumbosacral lipoma in 13 cases and was compared with conventional magnetic resonance imaging. In all cases, CISS images demonstrated the fine structure of the spinal cord and nerve roots, with good contrast between subarachnoid spaces and lipomas. The CISS findings were closely correlated with microscopic observation and electrophysiological investigation during surgery. In three of eight patients whose CISS images demonstrated abundant nerve roots embedded in the lipoma, untethering of the conus medullaris was not satisfactorily achieved. These three cases had obliteration of the subarachnoid space by the lipomas at the bilateral sides of the cord. In two cases with the vertebral misalignments scoliosis and lordosis, curvilinear reconstruction of CISS images along the spinal cord clearly demonstrated a neuroanatomy around the complicated lipomas. CONCLUSIONS: Using CISS imaging is useful for establishing precise neuroanatomical information around lumbosacral lipomas and for predicting the level of surgical difficulty. PMID- 16383254 TI - Tissue expansion for the delayed closure of large myelomeningoceles. AB - OBJECT: Numerous techniques for the soft-tissue closure of large myelomeningoceles have been described. When patients present for delayed repair beyond the neonatal period, the indications for surgery are both different and less immediate. METHODS: Six patients between 6 weeks and 3.5 years of age with defects ranging from 27 to 49 cm2 (mean 37 cm2) were referred to the authors' institution and were successfully treated by direct skin closure after tissue expansion. CONCLUSIONS: In this paper the authors describe the technique and advantages of expansion compared with those of locoregional flaps in this subgroup of patients with myelomeningoceles. PMID- 16383255 TI - Effect of trichostatin A, a histone deacetylase inhibitor, on glioma proliferation in vitro by inducing cell cycle arrest and apoptosis. AB - OBJECT: Trichostatin A (TSA) is a histone deacetylase inhibitor that causes growth inhibition of malignant cells. The authors' goal was to evaluate its effect on cell growth and cell cycle regulation in a large panel of glioma cell lines, as well as in human astrocytes, fibroblasts, and endothelial cells. METHODS: Cell growth in response to TSA was evaluated using a tetrazolium colorimetric assay and a clonogenic assay. Cell cycle effects were examined using flow cytometry. A DNA fragmentation assay was used to evaluate induction of apoptosis. Histone acetylation status and the expression of p21WAF1, phosphorylated retinoblastoma protein (Rb), poly(adenosine diphosphate-ribose) polymerase (PARP), and caspase-3 were studied using Western blot analysis. In the glioma cell lines, there was significant inhibition of cell growth and detection of increased levels of acetylated histones after TSA treatment. The mechanisms underlying the growth inhibition include cell cycle arrest at the G2/M phase and apoptosis induction. The expression of p21WAF1 was activated, with a temporally related decrease in levels of phosphorylated Rb. Apoptosis was preceded by detection of cleaved PARP and activated caspase-3. The effects of TSA were less pronounced or absent in human astrocytes, fibroblasts, and endothelial cells. CONCLUSIONS: The TSA caused inhibition of glioma cell growth by both cell cycle arrest and apoptosis. Cell cycle arrest was associated with an increase in p21WAF1 expression and a decrease in phosphorylated Rb. Apoptosis was mediated at least partly through the activation of caspase-3. Because of the differential effects in glioma cells compared with nonneoplastic cells, TSA may provide a novel strategy for achieving tumor growth inhibition and cytotoxicity. Further investigation is warranted. PMID- 16383256 TI - Diffusion tensor imaging of intraaxial tumors at the cervicomedullary and pontomedullary junctions. Report of two cases. AB - Brainstem gliomas are a heterogeneous group of lesions that account for 15% of all pediatric tumors of the central nervous system. Diagnosis and treatment planning for these tumors is based on the observation of Epstein and Farmer that the growth of lesions with low malignant potential is limited by the anatomical structures of the brainstem. Surgery is offered only to those patients with a high probability of harboring a low-grade tumor, because the attendant risk for significant morbidity outweighs the therapeutic benefit of debulking the tumor in cases of high-grade tumors. The authors report two cases that highlight the potential of diffusion tensor (DT) imaging to identify local white matter tracts in the pons, medulla, and cervical cord and to improve the preoperative assessment of low-grade gliomas. Preoperative DT imaging in both cases demonstrated that the white matter tracts were displaced by the bulk of the low grade tumors but were structurally preserved. Intraoperative and neurological findings were consistent with the preoperative interpretation of the DT images. These cases demonstrate that DT imaging is a useful method for visualizing the relationship between tumor and normal brainstem white matter architecture, as well as for improving the surgical evaluation and management of pediatric brainstem tumors. PMID- 16383257 TI - A C1-2 locked facet in a child with atlantoaxial rotatory fixation. Case report. AB - Conservative treatment is reported in a child with atlantoaxial rotatory fixation. Three-dimensional (3D) computerized tomography (CT) and magnetic resonance (MR) imaging demonstrated the degree of dislocation and alar ligament damage. A rigid cervical collar and muscle relaxant agents without any traction allowed full recovery. Control 3D CT scanning and MR imaging findings are reported. PMID- 16383259 TI - Fair compensation without litigation: addressing patients' financial need in disclosure. PMID- 16383258 TI - Chiari malformation Type I and a dolichoodontoid process responsible for sudden cardiorespiratory arrest. Case report. AB - The authors present the case of an 11-year-old girl who fell from standing height and experienced sudden cardiorespiratory arrest. A Chiari malformation Type I and a dolichoodontoid process were identified, and a brainstem contusion was found that correlated with the patient's clinical signs and symptoms. The authors hypothesize that the dolichoodontoid in a compromised craniocervical space created a contusion in the apposing brainstem at the time of head impact. The patient was left with permanent vocal cord paresis and spastic quadriparesis. Awareness of this rare occurrence is important in managing these congenital abnormalities and in assessing the risk of deterioration. PMID- 16383260 TI - Technology helps manage workers' compensation risks in the health care setting. PMID- 16383261 TI - Disclosure: what works now and what can work even better. AB - This three-part series on disclosure of unanticipated outcomes in health care is intended to provide an overview of the current thinking about disclosure and steps the organization can take to develop an approach to disclosure that is comprehensive and supportive of the needs of patients, families and providers. What should be apparent is that disclosure is not simply a requirement--it is a philosophy and part of a comprehensive approach to patient/family communication. PMID- 16383262 TI - Medical malpractice/ battery. Various causes of action available in 'ghost surgery' case. Meyers v. Epstein, 282 F. Supp. 2d 151 (S. D. N. Y. 2003). PMID- 16383263 TI - Vicarious liability. States differ in their treatment of agency. Estate of Carmine v. Sisters of Charity of Nazareth Health System, Inc., S. W. 3d, No. 2002 CA-001812-MR (Ct. App. Ky. Nov. 14, 2003); Roessler v. Novak, 858 S. E. 2d 1158 (Ct. App. Fla. 2003). PMID- 16383264 TI - Medical battery. Cause of action for battery may accrue when patient's wishes are disregarded. Hensley v. Scokin, S. W. 3d, No. M2002-00922-COA-R3-CV (Ct. App.Tenn. September 12, 2003). PMID- 16383265 TI - Long-term care. Agreement to arbitrate disputes may be unenforceable if it contravenes remedial rights. Romano v. Manor Care, Inc., 861 So. 2d 59 (Ct. App. 4th Dist. 2003). PMID- 16383266 TI - Mental health. Providers may be liable for releasing dangerous patients for lack of insurance. Bragg v. Valdez, 111 Cal. App. 4th 421, 3 Cal. Rptr.3d 804 (Ct. App. Cal 2nd Dist. 2003). PMID- 16383267 TI - Medical malpractice. Punitive damages allowed in the absence of compensatory damages in Alabama. Mobile Infirmary Medical Center v. Hodgen, So 3d, No. 1011932 (Ala. October 31, 2003). PMID- 16383268 TI - Medical training--DMETA's aim and organisation. PMID- 16383269 TI - The Managed Military Health System for Force Generation-project. AB - In summary, the Managed Military Health System for Force Generation will: Allow better understanding of health issues and illness patterns in the Armed Forces. Ensure optimum health and medical fitness of service personnel throughout their military careers by: Promoting health. Protecting health. Providing timely, properly co-ordinated, healthcare from military and civilian providers. In conjunction with DMICP, provide a seamless system of patient care and administration involving all healthcare providers. Enable all military personnel to make good health decisions. Ensure a smooth transition to NHS care on completion of Service. Improve the morale of those in the Defence Medical Services through working in a high quality organisation. PMID- 16383270 TI - The Defence Medical Operational Capability Project. PMID- 16383271 TI - Medical Operations. PMID- 16383272 TI - Fleet programming. PMID- 16383273 TI - Telemedicine in the Royal Navy. PMID- 16383274 TI - Medical support to littoral manoeuvre. AB - My personal view, after some 16 years away from 3 Commando Brigade, is that there has been a vast improvement in the medical infrastructure, logistics and communications at Unit level. The defined treatment timelines provide medical staff with much needed leverage in securing scarce SH assets and the casualty evacuation plan from Role 1 to Role 2 is cogent and workable. The Role 2 capability is much more flexible and robust and can now be projected forward to where it is needed. And, of course, we are now supported by a superb Role 3 facility afloat. All in all medical support to Lit M is in good shape, even if we are all still "dripping" about our tents! PMID- 16383275 TI - Medical manpower planning now and future. PMID- 16383276 TI - Dentistry in the Fleet. PMID- 16383277 TI - Impressions from sea--Medical Officer perspective. PMID- 16383278 TI - Primary casualty receiving facility [Argus on Operational Sea Training]. PMID- 16383279 TI - Impressions from sea--LMA perspective. PMID- 16383280 TI - An overview of the Institute of Naval Medicine. AB - In the time allotted to this presentation I have only been able to provide a brief outline of the type of work undertaken by INM. However, I hope I have been able to demonstrate that INM is indeed living up to its Mission Statement and motto 'e mari ad salutum'. PMID- 16383281 TI - Critical care retrieval revisited. AB - In 2000, the Journal of the Royal Naval Medical Service published an account of critical care retrieval experience written by Adrian Mellor and based on his time spent with CareFlight in Sydney, Australia. Having recently completed a term with the same organisation, I wanted to reiterate the usefulness of such an attachment. The purpose of this article is therefore to compare my experiences with those of Surgeon Commander Mellor and to provide an update on the work involved in critical care retrieval within New South Wales. PMID- 16383282 TI - The Defence Medical Library Service and military medicine. AB - The Defence Medical Library Service (DMLS) supports the clinical practice and career development of military health professionals across the world. Clinical governance and the need for medical knowledge to be evidence-based means the DMLS has a central role to play in support of defence medicine. The DMLS is important for enabling health professionals to make sense of the evidence-based pyramid and the hierarchy of medical knowledge. The Royal Centre for Defence Medicine (RCDM) in Birmingham is recognised as an international centre of excellence. The information, knowledge and research requirements of the RCDM will provide opportunities for the DMLS to support and engage with the academic community. PMID- 16383284 TI - [Everyday is a learning day]. PMID- 16383285 TI - [Facing difficulties, inheriting good traditions and marching to future]. PMID- 16383286 TI - [Opportunity and challenge of diagnostic pathology in China]. PMID- 16383287 TI - [Targeted therapy of neoplasms and pharmacopathology]. PMID- 16383288 TI - [Promoting development of pathology department]. PMID- 16383290 TI - [Basic and experimental pathology in China]. PMID- 16383289 TI - [History of the Chinese Journal of Pathology]. PMID- 16383291 TI - [Acuality and progress of pathologic techniques in China]. PMID- 16383292 TI - [Development of infection pathology in China]. PMID- 16383293 TI - [Pathology and genetics of disease and tumours of the lung, pleura in China]. PMID- 16383294 TI - [Liver pathology in China: retrospect and prospect]. PMID- 16383295 TI - [Kidney pathology in China]. PMID- 16383296 TI - [Lymphotic and hematological diseases pathology in China]. PMID- 16383297 TI - [Breast pathology in China]. PMID- 16383298 TI - [Progress and prospect of pediatric pathology in China]. PMID- 16383299 TI - [Detection of cyclin D1 mRNA by reverse transcription-polymerase chain reaction in paraffin-embedded tissues and its diagnostic significance for mantle cell lymphoma]. AB - OBJECTIVE: To investigate the feasibility of detecting cyclin D1 mRNA in paraffin embedded tissues by reverse transcriptase polymerase chain reaction (RT-PCR) and competitive RT-PCR and its diagnostic and differential diagnostic significance for mantle cell lymphoma (MCL). METHODS: Paraffin-embedded samples of 36 cases of MCL, 71 cases of other small B-cell lymphomas and 20 cases of lymphoid reactive hyperplasia as control group were retrieved from archival materials. Cyclin D1 protein and its mRNA was detected by EnVision and RT-PCR and competitive RT-PCR in all samples. House-keeping gene PGK was choosen as internal control. RESULTS: (1) Cyclin D1 protein was expressed in 27 of the 38 MCL (71.1%). No cyclin D1 expression was found in the control group. (2) PGK was detected in 103 of the 116 cases (88.8%) and also detected in 34 of 36 MCL cases (94.7%). (3) cyclin D1 mRNA was detected in 34 nodal mantle cell lymphoma cases by RT-PCR in paraffin embedded tissues. The positive rate of cyclin D1 mRNA was 94.4% in mantle cell lymphomas after exclusion of the 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia, except 1 case of B-SLL. Sequencing analysis showed that sequences were identical to cyclin D1. (4) Cyclin D1 mRNA overexpression was detected in 27 cases of nodal mantle cell lymphoma by competitive RT-PCR in paraffin-embedded tissues. The positive rate of cyclin D1 mRNA overexpression was 75.0% in mantle cell lymphomas after exclusion of 2 cases which were negative for both cyclin D1 mRNA and PGK. cyclin D1 mRNA overexpression was not detected in other nodal small B-cell lymphomas or lymphoid reactive hyperplasia. CONCLUSION: RT-PCR and competitive RT-PCR detection of cyclin D1 mRNA overexpression could be used for the diagnosis and differential diagnosis of mantle cell lymphoma in paraffin-embedded blocks. PMID- 16383300 TI - [Expressions of Axin and beta-catenin in non-small cell lung cancer]. AB - OBJECTIVE: To investigate the protein expression of Axin and beta-catenin, the exon 3 mutation status of beta-catenin and their clinicopathological correlations in non-small cell lung cancer (NSCLC). METHODS: A total of 100 NSCLC samples and their corresponding normal lung tissues were obtained from the patients undergoing surgery in the First Affiliated Hospital of China Medical University between 2001 and 2003. Protein expressions of Axin and beta-catenin were detected by immunohistochemistry. DNA sequence alterations of exon 3 of beta-catenin were investigated by polymerase chain reaction (PCR) and direct sequencing. RESULTS: A reduced membranous expression rate of beta-catenin was observed in 80.0% of the cases (80/100) along with a nuclear expression rate of 26.0% (26/100). There was a significant difference in beta-catenin expression between well and poorly differentiated NSCLCs. Well to moderately differentiated NSCLCs showed a reduced expression rate of 70.0% (35/50), in contrast to 90.0% (45/ 50) in poorly differentiated tumors (P = 0.012). Reduced beta-catenin expression rate was 87.3% (48/55) in cases with lymph node metastasis, in contrast to 71.1% (32/45) in cases without lymph node metastasis (P = 0.044). The positive expression rate of Axin was 48.0% (48/100). Well to moderately differentiated NSCLCs demonstrated a 60.0% positive expression rate of Axin (30/50), much higher than poorly differentiated tumors [36.0% (18/50), P = 0.016]. The positive expression rate of Axin in beta-catenin nuclear expressed NSCLCs was 15.4% (4/26), much lower than cases without beta-catenin nuclear expression [59.5% (44/74), P < 0.001]. Axin nuclear expression was found in two cases in this study, suggesting that it may function as a nuclear-cytoplasmic shuttling protein. PCR and direct sequencing failed to reveal any exon 3 mutation of beta-catenin gene. CONCLUSIONS: The reduced membranous expression of beta-catenin is associated with poorly differentiated and lymph node positive NSCLCs. The expression of Axin is inversely correlated with the degree of tumor differentiation and nuclear expression of beta-catenin. The exon 3 mutations do not contribute to the abnormal protein expression of beta-catenin in NSCLCs. PMID- 16383301 TI - [Prognostic significance of p21(WAF1) expression in osteosarcoma]. AB - OBJECTIVE: To investigate the expression of p21(WAF1) gene in human osteosarcoma and their relationships with clinicopathological parameters and prognostic value. METHODS: The p21(WAF1) gene mRNA and p21 protein expression in 45 osteosarcoma and 10 fibrous dysplasia of bone specimens were analyzed by in situ hybridization and immunohistochemistry respectively. RESULTS: (1) The positive rate of p21 protein expression in osteosarcoma was 17.7% (8/45). (2) The expression rate in high proliferation (4/10) significantly higher than that in low proliferation (4/35) osteosarcoma (chi2 = 4.34, P < 0.05). (3) The positive rate of p21(WAF1) mRNA expression in osteosarcoma was 42.2% (19/45). The expression rate in high proliferation (6/10) significantly higher than that in low proliferation (13/35) osteosarcoma (chi2 = 20.6, P < 0.01). (4) The survival time after operation of the patients with p21(WAF1) mRNA expression were higher than that of the patients with p21(WAF1) mRNA negative expression (P < 0.05). CONCLUSIONS: (1) With the increase in degree of malignancy, the expression of p21(WAF1) mRNA and p21 protein in osteosarcoma tend decrease. (2) The expression of p21(WAF1) mRNA has a definite value in judging prognosis in osteosarcoma. PMID- 16383302 TI - [Comparative study on the ultrastructures of radial and internal mammary arteries used for coronary artery bypass grafting]. AB - OBJECTIVE: The radial artery differs from internal mammary artery in its vascular biology and long-term patency after coronary artery bypass grafting (CABG). This study was designed to investigate their ultrastructural differences that may have implications in arterial remodeling and graft failure. METHODS: Thirty-four radial artery and 11 internal mammary artery samples were obtained from patients underwent CABG, and subjected to routine electron microscopic examination. A semi quantitative method was used to evaluate secretary endothelial cells, endothelial denudation, synthetic smooth muscle cells (SMCs), matrix accumulation, lipid deposition and medial submicroscopic calcification. RESULTS: Compared with internal mammary arteries, the radial arteries had more secretory endothelial cells (47.1%, 16/34 vs 27.2%, 3/ 11) and synthetic type SMCs in a background (14.4% vs 0.9%) that had more intimal lipid deposition and matrix accumulation (14.7%, 5/34 vs 9.1%, 1/11). Matrix vesicles and calcifications were frequently present in the media of both types of arteries. The calcifications, however, could not be visualized by routine histological stains, and therefore, named as submicroscopic calcification in this study. Fewer endothelial denudations were observed in the radial arteries, but no differences in medial lipid deposition and submicroscopic calcification were observed between these two types of arteries. The ultrastructural features and the arrangement of medial SMCs in radial arteries were similar to those of internal mammary arteries. CONCLUSIONS: Radial arteries have a higher SMC proliferative potential and more actively secretory status of endothelial cells, which may enhance the remodeling process and correlate with a decreased long-term patency. Better preservation of endothelial cells in radial arteries could be attributed to the "no touch" technique utilized in surgical harvesting. The significance of submicroscopic medial calcification during graft remodeling requires further investigations. PMID- 16383303 TI - [Establishment of a HPV-negative cell line derived from xenografted human laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To establish a human laryngeal carcinoma cell line unassociated with human papillomavirus (HPV). METHODS: Viable tissue of a well-differentiated laryngeal squamous cell carcinoma was obtained and tested negative for the presence of HPV by polymerase chain reaction. Minced tissue fragments were then transplanted subcutaneously into nude mice. After two successive passages, the tumor tissue was seeded into culture flasks and incubated in a medium containing 10% fetal bovine serum, insulin and epidermal growth factor. Tumor cell phenotype and molecular features were determined by various methods. RESULTS: A stable cell line, designated as Lscc-02, was successfully established after 86 culture passages. The cells grew as a monolayer with epithelioid features. The cell doubling time was approximately 39.1 hours. The human origin of the tumor cells was confirmed by karyotype analysis. The squamous epithelial phenotype was demonstrated by the immunopositivity of anti-cytokeratin antibodies and ultrastructural presence of tonofilaments and desmosomes. The malignant nature of the cells was documented by their clonal formation in soft-agar and tumorigenicity in nude mice. Lscc-02 cells expressed carcinoembryonic antigen (CEA) and were negative for HPV DNA. CONCLUSION: This newly established Lscc-02 laryngeal squamous cell carcinoma cell line may be a useful model for investigating laryngeal carcinoma unrelated to HPV infection, and the role of HPV in the progression of human laryngeal carcinoma. PMID- 16383304 TI - [Progress in the research of pathology and genetics of desmoid tumor]. PMID- 16383305 TI - [The World Health Organization and the Vienna classification of gastrointestinal epithelial neoplasia]. PMID- 16383306 TI - [Enteropathy T-cell lymphoma]. PMID- 16383307 TI - [World Health Organization classification of tumours, pathology and genetics of tumours of the lung]. PMID- 16383308 TI - Real-time monitoring of morphological changes in living cells by electronic cell sensor arrays: an approach to study G protein-coupled receptors. AB - G protein-coupled receptors (GPCRs) constitute important targets for drug discovery against a wide range of ailments including cancer, inflammatory, and cardiovascular diseases. Efforts are underway to screen selective modulators of GPCRs and also to deorphanize GPCRs with unidentified natural ligands. Most GPCR based cellular screens depend on labeling or recombinant expression of receptor or reporter proteins, which may not capture the true physiology or pharmacology of the GPCRs. In this paper, we describe a noninvasive and label-free assay for GPCRs that can be used with both engineered and nonengineered cell lines. The assay is based on using cell-electrode impedance to measure minute changes in cellular morphology as a result of ligand-dependent GPCR activation. We have used this technology to assay the functional activation of GPCRs coupled to different signaling pathways and have compared it to standard assays. We have used pharmacological modulators of GPCR signaling pathways to demonstrate the specificity of impedance-based measurements. Our data indicate that cell electrode impedance measurements offer a convenient, sensitive, and quantitative method for assessing GPCR function. Moreover, the noninvasive nature of the readout offers the added advantage of performing multiple treatments in the same well to study events such as desensitization and receptor cross-talk. PMID- 16383309 TI - Hadamard transform ion mobility spectrometry. AB - Traditionally, the spectrum acquired using ion mobility spectrometry (IMS) is an average of multiple experimental cycles. Each cycle is initiated by passing a short burst of ions into a drift tube containing a homogeneous electric field. Prior to starting the subsequent cycle, all ions in the system must arrive at the detector or spectral overlap may occur. To maximize resolution, the ion pulse admitted to the drift tube is small in relation to the total scan time with the unfortunate consequence of an inherently low duty cycle (approximately 1%). Offering an improved SNR through a 50% duty cycle, the Hadamard transform (HT) applied to ion mobility spectrometry represents a fresh alternative to signal averaged data acquisition. Initial results from measurements of amphetamine and cytochrome c samples indicate a 2-10-fold increase in SNR for the HT-IMS technique with no reduction in resolution. PMID- 16383310 TI - Lipid-bonded conducting polymer layers for a model biomembrane: application to superoxide biosensors. AB - Model biomembranes composed of poly-DATT/DGS/POPA and poly-DATT/DGS/CL were separately prepared on gold electrodes. A monolayer of 1,2-dioleoyl-sn-glycero-3 succinate (DGS) was covalently bonded onto electrochemically grown poly-(3,4 diamiono-2,2:5,2-terthiophene) (DATT) layers (thickness of approximately 300 nm; particle size of approximately 50 to 70 nm). The numbers of unit molecules of the poly-DATT layer and of the DGS immobilized onto the poly-DATT layers were 1.53 x 10(-7) and 1.56 x 10(-9) mol cm(-2), respectively, using a quartz crystal microbalance technique. The lipid bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphate (POPA) and cardiolipin (CL) were formed onto the poly-DATT/DGS layer using the Langmuir-Blodgett technique. The surface characterizations of each step were investigated by SEM, AFM, and XPS analyses. Cytochrome c (cyt c) was immobilized onto these model biomembranes through the charge interaction between the positive charges of cyt c and the negative charges of phosphate groups in CL or POPA lipids. At the POPA- and CL-modified biomembranes, the formal potentials of the redox couple of the immobilized cyt c were 0.22 and 0.23 V (vs Ag/AgCl), respectively. The redox reaction of the immobilized cyt c at the POPA- and CL modified biomembranes was quasireversible, and the electron-transfer rate constants were 0.121 s(-1) and 0.133 s(-1), respectively. The applicability of these cyt c immobilized bioimitation membranes as the biosensors was tested for the determination of superoxide. PMID- 16383311 TI - Titration force microscopy on supported lipid bilayers. AB - The use of chemically modified atomic force microscopy (AFM) probes allows us to measure the surface charges of supported planar lipid bilayers with high sensitivity through the force spectroscopy operation mode. By controlling the chemistry of the tip, we can perform a classical analytical chemistry titration where the titration agent is a weak acid (attached to the AFM tip) with the particularity of being performed in surface rather than in solution and, especially, at the nanometric scale. Thus, the AFM tip acts as a real "nanosensor". The approaching curves of the force plots reveal that electrostatic interactions between the tip and the supported membrane play a key role. Besides, the plot of the adhesion force (measured from the retracting curve of the force plots) versus pH displays a nonsigmoidal shape with a peak in the adhesion force attributed to high-energy hydrogen bonds. One of these peaks corresponds to the pKa of the surface under study and the other to the pKa of the titrating probe attached to the tip. PMID- 16383312 TI - Development of an enzyme-linked immunosorbent assay for the determination of the linear alkylbenzene sulfonates and long-chain sulfophenyl carboxylates using antibodies generated by pseudoheterologous immunization. AB - ELISA methods have been developed for screening contamination of water resources by linear alkyl benzene sulfonates (LAS) or the most immediate degradation products, the long chain sulfophenyl carboxylates, SPCs. The assay uses antibodies raised through pseudoheterologous immunization strategies using an equimolar mixture of two immunogens (SFA-KLH and 13C(13)-SPC-KLH) prepared by coupling N-(4-alkylphenyl)sulfonyl-3-aminopropanoic acid (SFA) and p-(1-carboxy 13-tridecyl)phenylsulfonic acid (13C(13)-SPC) to keyhole limpet hemocyanin (KLH). The immunizing haptens have been designed to address recognition versus two different epitopes of the molecule. The SFA hapten maximizes recognition of the alkyl moiety while preserving the complexity of the different alkyl chains present in the LAS technical mixture. The 13C(13)-SPC hapten addresses recognition of the common and highly antigenic phenylsulfonic group. The antisera raised using this strategy have been shown to be superior to those obtained through homologous immunization procedures using a single substance. By using an indirect ELISA format, LAS and long-chain SPCs can be detected down to 1.8 and 0.2 microg L(-1), respectively. Coefficients of variation of 6 and 12% within and between assays, respectively, demonstrate immunoassay reproducibility. The assay can be used in media with a wide range of pH and ionic strength values. Preliminary experiments performed to assess matrix effects have demonstrated the potential applicability of the method as a screening tool to assess contamination by these types of surfactants in natural water samples. PMID- 16383313 TI - Rapid electrochemiluminoimmunoassay of human C-reactive protein at planar disposable oxide-coated silicon electrodes. AB - Electrochemiluminescence (ECL) of aromatic Tb(III) chelates at thin insulating film-coated electrodes provides a means for extremely sensitive detection of Tb(III) chelates and also of biologically interesting compounds if these chelates are used as labels in bioaffinity assays. The suitability of silicon electrodes coated with thermally grown silicon dioxide film as disposable working electrodes in sensitive time-resolved ECL measurements is demonstrated, and a rapid electrochemiluminoimmunoassay (ECLIA) of human C-reactive protein (hCRP) is described. Tb(III) chelate labels can be detected almost down to picomolar level, and the calibration curve of these labels covers more than 6 orders of magnitude of chelate concentration. The calibration curve of the present immunometric hCRP assay was found to be linear over a wide range, approximately 4 orders of magnitude of hCRP concentration, the detection limit of the protein being 0.3 ng mL(-1) (mean background + 2SD) on CV values of about 10-30%, depending on the immunoassay incubation time. In the ECLIA measurements, different incubation times were tested from 15 min (giving above-mentioned performance) to as short as only 2 min, which still gave successful results with approximately 20,000 times better detection limit levels than traditional commercial assay methods. During the ECLIA process, also the Si electrode surface morphology was also investigated by atomic force microscope monitoring. PMID- 16383315 TI - Characterization of gas-phase molecular interactions on differential mobility ion behavior utilizing an electrospray ionization-differential mobility-mass spectrometer system. AB - Differential mobility spectrometry (DMS) is a rapidly advancing technology for gas-phase ion separation. The interfacing of DMS with mass spectrometry (MS) offers potential advantages over the use of mass spectrometry alone. Such advantages include improvements to mass spectral signal/noise ratios, orthogonal/complementary ion separation to mass spectrometry, enhanced ion and complexation structural analysis, and potential for rapid analyte quantitation. The introduction of a new ESI-DMS-MS system and its utilization to aid in the understanding of DMS separation theory is described. A current contribution to DMS separation theory is one of an association/dissociation process between ions/molecules in the gas phase during the differential mobility separation. A model study was designed to investigate the molecular dynamics and chemical factors influencing the theorized association/dissociation process, and the mechanisms by which these gas-phase interactions affect an ion's DM behavior. Five piperidine analogues were selected as model analytes, and three alcohol drift gas dopants/modifiers were used to interrogate the analyte ions in the gas phase. Two proposed DMS separation mechanisms, introduced as Core and Facade, corresponding to strong and weak attractions between ions/molecules in the gas phase, are detailed. The proposed mechanisms provide explanation for the observed changes in analyte separation by the various drift gas modifiers. Molecular modeling of the proposed mechanisms provides supportive data and demonstrates the potential for predictive optimization of analyte separation based on drift gas modifier effects. PMID- 16383314 TI - Central limit theorem as an approximation for intensity-based scoring function. AB - In this paper, we present an intensity-based probability function to identify peptides from tandem mass spectra and amino acid sequence databases. The function is an approximation to the central limiting theorem, and it explicitly depends on the cumulative product ion intensities, number of product ions of a peptide, and expectation value of the cumulative intensity. We compare the results of database searches using the new scoring function and scoring functions from earlier algorithms, which implement hypergeometric probability, Poisson's model, and cross-correlation scores. For a standard protein mixture (tandem mass spectra generated from the mixture of five known proteins), we generate receiver operating curves with all scoring schemes. The receiver operating curves show that the shared peaks count-based probability methods (like Poisson and hypergeometric models) are the most specific for matching high-quality tandem mass spectra. The intensity-based (central limit model) and intensity-modeled (cross-correlation) methods are more sensitive when matching low-quality tandem mass spectra, where the number of shared peaks is insufficient to correctly identify a peptide. Cross-correlation methods show a small advantage over the intensity-based probability method. PMID- 16383316 TI - Direct writing of metal nanoparticle films inside sealed microfluidic channels. AB - Herein we demonstrate the ability to pattern Ag nanoparticle films of arbitrary geometry inside sealed PDMS/TiO2/glass microfluidic devices. The technique can be employed with aqueous solutions at room temperature under mild conditions. A 6 nm TiO2 film is first deposited onto a planar Pyrex or silica substrate, which is subsequently bonded to a PDMS mold. UV light is then exposed through the device to reduce Ag+ from an aqueous solution to create a monolayer-thick film of Ag nanoparticles. We demonstrate that this on-chip deposition method can be exploited in a parallel fashion to synthesize nanoparticles of varying size by independently controlling the solution conditions in each microchannel in which the film is formed. The film morphology was checked by atomic force microscopy, and the results showed that the size of the nanoparticles was sensitive to solution pH. Additionally, we illustrate the ability to biofunctionalize these films with ligands for protein capture. The results indicated that this could be done with good discrimination between addressed locations and background. The technique appears to be quite general, and films of Pd, Cu, and Au could also be patterned. PMID- 16383317 TI - Theory and use of the pseudophase model in gas-liquid chromatographic enantiomeric separations. AB - The theory and use of the "three-phase" model in enantioselective gas-liquid chromatography utilizing a methylated cyclodextrin/polysiloxane stationary phase is presented for the first time. Equations are derived that account for all three partition equilibria in the system, including partitioning between the gas mobile phase and both stationary-phase components and the analyte equilibrium between the polysiloxane and cyclodextrin pseudophase. The separation of the retention contributions from the achiral and chiral parts of the stationary phase can be easily accomplished. Also, it allows the direct examination of the two contributions to enantioselctivity, i.e., that which occurs completely in the liquid stationary phase versus the direct transfer of the chiral analyte in the gas phase to the dissolved chiral selector. Six compounds were studied to verify the model: 1-phenylethanol, alpha-ionone, 3-methyl-1-indanone, o (chloromethyl)phenyl sulfoxide, o-(bromomethyl)phenyl sulfoxide, and ethyl p tolylsulfonate. Generally, the cyclodextrin component of the stationary phase contributes to retention more than the bulk liquid polysiloxane. This may be an important requirement for effective GC chiral stationary phases. In addition, the roles of enthalpy and entropy toward enantiorecognition by this stationary phase were examined. While enantiomeric differences in both enthalpy and entropy provide chiral discrimination, the contribution of entropy appears to be more significant in this regard. The three-phase model may be applied to any gas liquid chromatography stationary phase involving a pseudophase. PMID- 16383318 TI - Coupled surface-enhanced Raman spectroscopy and electrical conductivity measurements of 1,4-phenylene diisocyanide in molecular electronic junctions. AB - Probing the structure of molecules in a metal-molecule-metal junction under an applied voltage is critical for understanding molecular electron transport properties. We present an approach that allows recording surface-enhanced Raman spectra simultaneously with electrical measurements of a monolayer of molecules in molecular electronic junctions. 1,4-Phenylene diisocyanide in two different types of junctions was used to illustrate the approach. The results show that the molecular integrity was intact in the molecular junctions and under the applied bias. The monolayer sensitivity of the approach provides a new powerful tool for characterizing molecular structure in a molecular electronic junction. PMID- 16383319 TI - Efficient and specific trypsin digestion of microgram to nanogram quantities of proteins in organic-aqueous solvent systems. AB - Mass spectrometry-based identification of the components of multiprotein complexes often involves solution-phase proteolytic digestion of the complex. The affinity purification of individual protein complexes often yields nanogram to low-microgram amounts of protein, which poses several challenges for enzymatic digestion and protein identification. We tested different solvent systems to optimize trypsin digestions of samples containing limited amounts of protein for subsequent analysis by LC-MS-MS. Data collected from digestion of 10-, 2-, 1-, and 0.2-microg portions of a protein standard mixture indicated that an organic aqueous solvent system containing 80% acetonitrile consistently provided the most complete digestion, producing more peptide identifications than the other solvent systems tested. For example, a 1-h digestion in 80% acetonitrile yielded over 52% more peptides than the overnight digestion of 1 microg of a protein mixture in purely aqueous buffer. This trend was also observed for peptides from digested ribosomal proteins isolated from Rhodopseudomonas palustris. In addition to improved digestion efficiency, the shorter digestion times possible with the organic solvent also improved trypsin specificity, resulting in smaller numbers of semitryptic peptides than an overnight digestion protocol using an aqueous solvent. The technique was also demonstrated for an affinity-isolated protein complex, GroEL. To our knowledge, this report is the first using mass spectrometry data to show a linkage between digestion solvent and trypsin specificity. PMID- 16383320 TI - Use of porous membranes modified with polyelectrolyte multilayers as substrates for protein arrays with low nonspecific adsorption. AB - Coating of substrates with polyelectrolyte multilayers terminated with poly(acrylic acid) (PAA) followed by activation of the free -COOH groups of PAA provides a surface that readily reacts with amine groups to allow covalent immobilization of antibodies. The use of this procedure to prepare arrays of antibodies in porous alumina supports facilitates construction of a flow-through system for analysis of fluorescently labeled antigens. Detection limits in the analysis of Cy5-labeled IgG are 0.02 ng/mL because of the high surface area of the alumina membrane, and the minimal diameter of the substrate pores results in binding limited by kinetics, not mass transport. Moreover, PAA-terminated films resist nonspecific protein adsorption, so blocking of antibody arrays with bovine serum albumin is not necessary. These microarrays are capable of effective analysis in 10% fetal bovine serum. PMID- 16383321 TI - Metal nanoparticle deposition for TOF-SIMS signal enhancement of polymers. AB - A novel technique for improved time-of-flight secondary ion mass spectra of polymer ions is presented. This technique is a simple preparatory method, which involves deposition of a submonolayer coverage of metal nanoparticles on the surface of a polymer sample enabling an overall increase in characteristic polymer ions. This procedure gives spectra with enhanced intensity, a larger number of characteristic polymer peaks, and peaks of higher mass. Both Au and Ag nanoparticles were employed to facilitate the ionization of the polymer characteristic secondary ions. Moreover, these experiments demonstrate that the nanoparticles allow localization of high-mass fragment ions during imaging experiments utilizing focused ion beams. In general, we show that the metal nanoparticle deposition method is effective for time-of-flight secondary ion mass spectrometry examination of polymers. PMID- 16383322 TI - Air-stable G protein-coupled receptor microarrays and ligand binding characteristics. AB - This paper described novel strategies to achieve air-stable G protein-coupled receptor (GPCR) microarrays and the uses of the microarrays for ligand profiling. Specifically, GPCR cell membrane fragments were suspended in a buffered solution containing bovine serum albumin (BSA) and disaccharide sucrose or trehalose and used for fabricating GPCR microarrays. During the array fabrication and postfabrication processes, BSA molecules were found to effectively form packed layer(s) surrounding the GPCR membranes immobilized onto the predetermined printing area, thereby stabilizing the membrane microspots. The use of disaccharides was shown to protect the integrity and functionality of GPCR microarrays from the typical deterioration of the membranes when fabricated and stored under dry conditions. To utilize the ability of fluorescence technology for multichannel detection as well as to maximize the capability of GPCR microarrays for multiplexed binding assays, several fluorescently labeled ligands were synthesized and optimized for multiplexing binding assays. A schematic microarray of five GPCRs had been used as a model for characterizing the association and dissociation rate constants of labeled ligands binding to their respective receptors in the microarrays. Interestingly, distinct receptor-ligand interactions exhibited different dependence on the type of pH reagent as well as the species and concentration of cations used in a binding assay buffered solution. The potential mechanisms and implications for the uses of air-stable GPCR microarrays were discussed. PMID- 16383323 TI - Chemical speciation of iron in seawater by cathodic stripping voltammetry with dihydroxynaphthalene. AB - The chemical speciation of iron in seawater is determined by cathodic stripping voltammetry using 2,3-dihydroxynaphthalene (DHN) as adsorptive and competing ligand. The optimized conditions include a DHN concentration of 0.5-1 microM, seawater at its original pH of 8, and equilibration overnight. The alpha coefficient for DHN (=[FeDHN]/[Fe']) was calibrated against EDTA giving values of 166 for 0.5 microM DHN and 366 at 1 microM DHN and a value of 8.51 +/- 0.07 for log K'(Fe'DHN). The dissociation of the natural iron species FeL was found to have a characteristic reaction time of 50 min, indicating that titrations should be equilibrated overnight rather than the shorter periods sometimes used onboard ship. The method was applied to samples from the Pacific giving ligand concentrations of 1.1 and 1.6 nM for deep and surface waters, respectively, with an average value for log K'(FeL) of 11.9 +/- 0.3 compared to a value of 11.5 for the siderophore deferoxamine. The results are similar to those obtained previously for similar samples, but the new method has much greater sensitivity for iron than previous methods, leading to lower limits of detection and shorter analysis time. PMID- 16383324 TI - Matrix-assisted laser desorption/ionization high-energy collision-induced dissociation of steroids: analysis of oxysterols in rat brain. AB - Neutral steroids have traditionally been analyzed by gas chromatography/mass spectrometry (GC/MS) after necessary derivatization reactions. However, GC/MS is unsuitable for the analysis of many conjugated steroids and those with unsuspected functional groups. Here we describe an alternative analytical method specifically designed for the analysis of oxosteroids and those with a 3beta hydroxy-delta5 or 5alpha-hydrogen-3beta-hydroxy structure. Steroids were derivatized with Girard P (GP) hydrazine to give GP hydrazones, which are charged species and readily analyzed by matrix-assisted laser desorption/ionization mass spectrometry. The resulting [M]+ ions were then subjected to high-energy collision-induced dissociation on a tandem time-of-flight instrument. The product ion spectra give structurally informative fragment ion patterns. The sensitivity of the analytical method is such that steroid structures can be determined from low-picogram (low-femtomole) amounts of sample. The utility of the method has been demonstrated by the analysis of oxysterols extracted from rat brain. PMID- 16383325 TI - Surface binding affinity measurements from order transitions of lipid membrane coated colloidal particles. AB - Lipid bilayers can be assembled onto the surface of colloidal silica particles to form a continuous and fluid supported membrane coating. In this configuration, the collective behavior of the colloidal dispersion is governed by interactions between particles and exhibits a sensitive dependency on chemical features of the membrane surface. Protein binding to membrane surface receptors can trigger macroscopic changes in the colloidal order, which provides a label-free readout of such binding events. Here, the relationship between order in the colloidal dispersion and the surface concentration of bound protein is characterized quantitatively in terms of the radial pair distribution function. Using parallel fluorescence measurements for comparison, we construct a scalar measure of the distribution function that exhibits linear proportionality with surface protein binding. This is used to determine binding affinity based only on observations of the colloidal distribution. PMID- 16383326 TI - Rapid chemical digestion of small acid-soluble spore proteins for analysis of Bacillus spores. AB - A method for the rapid identification of Bacillus spores is proposed, based on the selective release and chemical digestion of small, acid-soluble spore proteins (SASPs). Microwave-assisted acid hydrolysis of SASPs from B. anthracis str. Sterne and B. subtilis str. 168 was accomplished in a single step requiring only 90 s of heating. The peptide products of the chemical digestion were identified by postsource decay sequencing with a MALDI-TOF-MS equipped with a curved-field reflectron. The specificity of the observed SASP peptides was evaluated using a cross-species sequence search. The incomplete nature of the acid digestion under these conditions allowed detection of the digest products along with the proteins from which they originated, which increased species identification confidence. The feasibility of this approach for the rapid identification of Bacillus species was further demonstrated by analyzing a mixture of B. subtilis str. 168 and B. anthracis str. Sterne spores. PMID- 16383327 TI - Unified equation for access to rate constants of first-order reactions in dynamic and on-column reaction chromatography. AB - A unified equation to evaluate elution profiles of reversible as well as irreversible (pseudo-) first-order reactions in dynamic chromatography and on column reaction chromatography has been derived. Rate constants k1 and k(-1) and Gibbs activation energies are directly obtained from the chromatographic parameters (retention times tR(A) and tR(B) of the interconverting or reacting species A and B, the peak widths at half-height wA and wB, and the relative plateau height h(p)), the initial amounts A0 and B0 of the reacting species, and the equilibrium constant K(A/B). The calculation of rate constants requires only a few iterative steps without the need of performing a computationally extensive simulation of elution profiles. The unified equation was validated by comparison with a data set of 125,000 simulated elution profiles to confirm the quality of this equation by statistical means and to predict the minimal experimental requirements. Surprisingly, the recovery rate from a defined data set is on average 35% higher using the unified equation compared to the evaluation by iterative computer simulation. PMID- 16383328 TI - Characterization of various analytes using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 2-[(2E)-3-(4-tert butylphenyl)-2-methylprop-2-enylidene]malononitrile matrix. AB - 2-[(2E)-3-(4-tert-Butylphenyl)-2-methylprop-2-enylidene]malononitrile (DCTB) is a nonpolar, aprotic matrix and was used in the analysis of a variety of compounds by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The classes of compounds include coordination compounds, organometallics, conjugated organic compounds (including porphyrins and phthalocyanines), carbohydrates, calixarenes, and macrocycles. For some samples, comparisons are made with spectra acquired with the use of 1,8,9 trihydroxyanthracene (dithranol), 2,5-dihydroxybenzoic acid, and 2,4,6 trihydroxyacetophenone matrixes. Traditionally, the majority of these compounds would have been analyzed by fast-atom bombardment (FAB), liquid secondary ion mass spectrometry (LSIMS), or electrospray techniques, but this work shows that MALDI-TOFMS using DCTB has advantages over these techniques, particularly FAB and LSIMS. Certain limitations of DCTB are noted, for example, in the analysis of water-soluble compounds such as peptides, proteins, and oligonucleotides, and good working practices for the use of the matrix are also outlined. PMID- 16383329 TI - Quantitating the statistical distribution of deuterium incorporation to extend the utility of H/D exchange MS data. AB - Measuring the statistical distribution of deuterium incorporated into enzymatically derived peptide fragments provides a valuable dimension to hydrogen/deuterium exchange mass spectrometry data. In this paper, we will discuss our improvement to the linear least-squares method for determining this distribution, through the addition of "zeroes" to the end of the deuterated isotopic envelope, to partially compensate for data truncation due to finite instrumental signal-to-noise ratios. The value of the distribution is demonstrated in a simple experimental example, where the linearity between average deuteration and percent D2O used to label test peptides hides a more complex relationship between the site-labeling probability and the total number of sites. This method offers the opportunity to resolve cases where a single peptide experiences distinct, independent biochemical states with each bearing a unique average deuteration; this can occur when a protein is modified to substoichiometric levels. From the experimentally determined distribution of a heterogeneously deuterated peptide, it was possible to extract the average deuteration of each component of the mixture. PMID- 16383330 TI - In vivo near-infrared spectroscopy of rat skin tissue with varying blood glucose levels. AB - We have performed in vivo measurements of near-infrared rat skin absorption in the 4000-5000-cm(-1) spectral range (2.0-2.5-microm wavelength) during a glucose clamp experiment in order to identify the presence of glucose-specific spectral information. Spectra were collected during an initial 3-h period where the animal's blood glucose concentration was held at its normal value. The blood glucose level was then increased above 30 mM by venous infusion of glucose and held for 2 h, after which it was allowed to return to normal. Spectra were recorded continuously during the procedure and are analyzed to identify spectral changes associated with changes in glucose concentration. Because the change in absorbance due to an increase in glucose concentration is small compared to changes due to other variations (e.g., the thickness of the skin sample), a simple subtraction of absorbance spectra from the hyperglycemic and euglycemic phases is not instructive. Instead, a set of principal components is established from the euglycemic period where the glucose concentration is constant. We then examine the change in absorbance during the hyperglycemic period that is orthogonal to these principal components. We find that there are significant similarities between these orthogonal variations and the net analyte signal of glucose, which suggests that glucose spectral information is present. The analysis described here provides a procedure by which the analytical significance of a multivariate calibration can be evaluated. PMID- 16383331 TI - From micro to nano: analysis of surface-enhanced resonance Raman spectroscopy active sites via multiscale correlations. AB - Effective correlation of data from a number of analytical techniques over length scales spanning several orders of magnitude is required to more fully investigate the active sites on silver nanoparticles that are responsible for surface enhanced resonance Raman scattering (SERRS). In this paper, a method is presented that uses fluorescent beads as optical markers to allow direct correlation between a SERRS/fluorescence map and a transmission electron microscope (TEM) collage of the same area. Factors influencing the accuracy of the technique include the flatness of the substrate, the size of the fluorescent beads, and the strength of the signal from the fluorescent beads. When the effect of each of these factors on the technique is addressed, a simple and accurate correlation between the optical spectroscopy and the electron microscopy is achieved. A statistically significant number of particles can then be easily and reliably located and characterized at both optical limits, by SERRS, and with subnanometer resolution in the high-resolution TEM. Examples of HRTEM images and the locations of these particles within the SERRS map/TEM collage are presented. Our findings reveal that the relative SERRS activity of single particles is very low compared to dimers and larger aggregates of particles. The relative activity of dimers is estimated to be 12.4 times greater than single particles, and as the number of particles in the aggregate increase, the relative SERRS activity also increases. The relative SERRS activities of single particles/dimers/trimers/aggregates of 4 9 particles/aggregates of 10-20 are estimated to be 1/12.4/15.6/23.2/43. PMID- 16383332 TI - Measurement of enzyme activity in single cells by voltammetry using a microcell with a positionable dual electrode. AB - The electrochemical single-cell analysis for enzyme activity was developed using microcells on a microcell array coupled with a positionable dual microelectrode. The microcell array with the nanoliter-scale microcells was constructed using simple chemical etching without photolithographic techniques. The positionable dual microelectrodes consisted of the nanometer-to-micrometer-radius Au disk working electrode and a approximately 80-microm-radius Ag/AgCl reference electrode. Peroxidase was chosen as the model enzyme. Factors that concern electrochemical single-cell analysis in microcells such as solution evaporation, interference of soluble oxygen, electrode size, solution volume, and electrode fouling were investigated and discussed. The 20 or 100 nL of detection volume was found to be suitable for peroxidase determination in single neutrophils or single acute promyelocytic leukemia cells without interference from intracellular macromolecules and electrode fouling, when the dual electrode with a 10-microm radius Au disk working electrode was used. Cells were perforated with digitonin before transferring them into the microcells, to lyse cells easily. The perforated cells were transferred into the microcells by pushing a microscope slide on a drop of the cell suspension on the microcell array. After a single cell in the microcell was lysed using a freeze-thawing technique and allowed to dry, physiological buffer saline containing 2.0 x 10(-3) mol/L hydroquinone and 2.0 x 10(-3) mol/L H2O2 as the substrates of the enzyme-catalyzed reaction was added. The microcell array was positioned in a constant-humidity chamber to prevent evaporation. Then the dual electrode was inserted into the microcell by means of a scanning electrochemical microscope and the product benzoquinone of the enzyme-catalyzed reaction was voltammetrically detected. Peroxidase activity could be quantified using the steady-state current on the voltammogram after subtracting the blank and using the calibration curve. PMID- 16383333 TI - Simultaneous determination of pKa and lipophilicity by gradient RP HPLC. AB - High-performance methods of testing of drug candidates for properties of pharmacokinetics and pharmacodynamics importance, in particular lipophilicity and acidity, are necessary to overcome innovation stagnation in the pharmaceutical industry. Reversed-phase high-performance liquid chromatography (RP HPLC) might be a unique tool for the determination of both pKa and the apparent (pH dependent) partition coefficient, applicable in high-throughput analysis of multicomponent mixtures, e.g., samples originating from automated synthesis. In this work, the pH/organic modifier gradient RP HPLC is presented as a means of simultaneous determination of an analyte's acidity and lipophilicity. The approach consists of retention measurements in a series of methanol gradient runs differing in pH range and duration of the gradient. Two different models of the influence of pH on retention in organic modifier gradient RP HPLC are compared regarding the quality of the simultaneously determined lipophilicity and dissociation constants. Advantages of the proposed approach over currently employed procedures are that it can be applied to compound mixtures, it requires only minute amounts of substances, and pKa values can be determined in the range 3-10 units and lipophilicity in the range 0-7 units. Verification of the reliability of the parameters determined by the new method was demonstrated on a series of 93 acidic and basic drug analytes. PMID- 16383334 TI - Chromatographic purification for the determination of dissolved silicon isotopic compositions in natural waters by high-resolution multicollector inductively coupled plasma mass spectrometry. AB - A procedure is described for accurate Si isotope ratio measurements by multicollector inductively coupled plasma mass spectrometry (MC-ICPMS). Dissolved silicon was preconcentrated and separated from other elements present in natural surface waters using anion-exchange chromatography. The optimized procedure provides virtually complete elimination of major inorganic constituents while maintaining Si recovery in excess of 97%. High-resolution capabilities of MC ICPMS used in this study allow interference-free measurements of 28Si and 29Si isotopes using conventional solution nebulization sample introduction without aerosol desolvation. Owing to the magnitude of polyatomic ion contributions in the region of mass 30, mostly from 14N16O+, measurements of the 30Si isotope can be affected by tailing of the interference signals, making exact matching of analyte and nitric acid concentrations in all measurement solutions mandatory. Isotope abundance ratio measurements were performed using the bracketing standards approach and on-line correction for mass-bias variations using an internal standard (Mg). Uncertainties, expressed as 95% confidence intervals, for replication of the entire procedure are better than +/-0.18/1000 for delta29Si and +/-0.5/1000 for delta30Si. For the first time with MC-ICPMS, the quality of Si isotope abundance ratio measurements could be verified using a three-isotope plot. All samples studied were isotopically heavier than the IRMM-018 Si isotopic reference material. PMID- 16383335 TI - Recognition of proteins by crystallization patterns in an array of reporter solution microdroplets. AB - A new technique is described for specific recognition of protein analytes by observing protein-induced changes in the drying/crystallization patterns (DCP) of an array of microdroplets containing solutions of different reporter substances. Recognition is based on a difference in interaction of the protein analyte crystalline elements (planes, edges, defects, etc.) in the growing reporter crystals. Using a set of natural L-amino acids as reporters and denoting the amino acid solutions displaying substantial protein-induced changes in the DCP as "1" and those that show no or small changes as "0", a digital binary code was determined for several proteins at multiple concentrations. It was demonstrated that globular proteins can be reliably identified using this code as a "signature" when only 2-100 ng of protein was added to amino acid microdroplets. PMID- 16383336 TI - Fabrication and characterization of nanostructured Pd hydride pH microelectrodes. AB - Novel pH microsensors were made by electrodepositing mesoporous Pd films onto Pt microdisks, electrochemically loading the films with hydrogen to form the alpha+beta Pd hydride phase, and then switching to the potentiometric mode to monitor pH. To create a nanostructure, the films were deposited within a molecular template formed by the self-assembly of surfactant molecules, a technique known as true liquid crystal templating. The films retain the micrometer size of the Pt microdisk but offer electroactive areas up to 900 times larger. Optimum hydrogen loading conditions were determined, and the mesoporous Pd microdisks were found to have excellent potentiometric properties. From pH 2 to 12, their potential was Nernstian, highly reproducible, very stable (+/-1.2 mV over 2 h), and without hysteresis. Their response time was better than 1 s. However, the presence of oxygen reduced their lifetime significantly, thereby requiring frequent reloading. These microelectrodes do not require calibration before and after measurements, a procedure normally essential for potentiometric pH microsensors. To our knowledge, these are the first results where nanostructured materials made by the true liquid crystal templating method have been used in the potentiometric mode; moreover, these are the first results demonstrating the application of nanostructured microdisks in the potentiometric mode. PMID- 16383337 TI - Surfactant-enhanced desorption/ionization on silicon mass spectrometry. AB - Perfluorinated surfactants are demonstrated to dramatically enhance desorption/ionization on fluorinated silicon (DIOS) mass spectrometry. Perfluorooctanesulfonic acid improved the signal-to-noise ratio of tryptic digests and gave a 3-fold increase in the number of peptides identified. Similar results were also obtained using perfluoroundecanoic acid; yet among the seven different surfactants tested, controls such as nonfluorinated sodium dodecyl sulfate or fluorinated molecules with minimal surfactant activity did not enhance the signal. The same surfactants also enhanced the DIOS-MS signal of amino acids, carbohydrates, and other small organic compounds. The signal enhancement may be facilitated by the high surface activity of the perfluorinated surfactants on the fluorinated silicon surfaces allowing for a higher concentration of analyte to be absorbed. PMID- 16383338 TI - Transient signal analysis using complementary metal oxide semiconductor capacitive chemical microsensors. AB - This work explores the possibility to discriminate analytes based on their nonequilibrium signals in polymer-coated capacitive chemical microsensors. The analyte uptake in the chemically sensitive polymer layers of 3-7-microm thickness has been analyzed using a diffusion model and the dynamic sensor response data. The shapes of the response profiles have been calculated analytically. Despite the simplifications in the model, the observed transient signal profiles could be described accurately. Comparison of the measured diffusion coefficients (on the order of 10(-12) m2/s) with literature values measured at similar concentration levels showed good agreement. Concentration-independent diffusion coefficients for several analyte/polymer combinations (poly(etherurethane)/all analytes; poly(epichlorohydrin)/alcohols) as well as slightly concentration-dependent diffusion coefficients (poly(epichlorohydrin)/toluene or ethyl cellulose/toluene) have been found in the investigated concentration range of tens to hundreds of pascals gas-phase partial pressure. The diffusion times of water and the first aliphatic monohydric alcohols in the polymers are strongly correlated to their molecular size. The discrimination of these substances based on dynamic sensor data of a single sensor could be demonstrated. In particular, the analysis of mixtures of analytes with similar chemical behavior (water/ethanol or methanol/ethanol) by means of analyzing the response profile of single-exposure steps or by applying a series of decreasingly long alternating target gas exposure and carrier gas exposure steps has been performed. PMID- 16383339 TI - Quantification of peptides for the monitoring of protease-catalyzed reactions by matrix-assisted laser desorption/ionization mass spectrometry using ionic liquid matrixes. AB - Ionic liquid matrixes (ILM) have been shown to allow very homogeneous sample preparations, facilitating relative quantifications using internal standards in matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). In the present work, the ability to perform quantifications of peptides without using internal standards in these matrixes was investigated. Linear correlations between peptide amount and signal intensities could be observed when increased molar matrix-to-analyte ratios were applied. The dynamic range of linearity was approximately 1 order of magnitude. The method was applied successfully to monitor the time-dependent evolution of substrates and products in trypsin catalyzed digests of single peptides and peptide mixtures. Thus, ionic liquid matrixes allow quantitative MALDI-MS without the need for internal standards, making the method a suitable tool for the fast screening of new enzymes or the search for substrates or inhibitors. PMID- 16383340 TI - Substrates for direct imaging of chemically functionalized SiO2 surfaces by transmission electron microscopy. AB - A significant challenge in materials characterization is the determination of the structure of nanoparticle assemblies that have been deposited on solid substrates, such as SiO2. The best method for obtaining quantitative information about structure, size, and spacing on the nanometer-length scale is TEM; however, commercially available TEM grids offer a limited range of substrate materials. In addition, the compositions of these grids do not permit much chemical processing. Here we describe silicon-based grids with electron-transparent SiO2 windows suitable for use as substrates for high-resolution TEM that can be easily fabricated using standard silicon microfabrication techniques. These grids are physically and chemically robust and exhibit the same surface chemistry and chemical stability as an oxide grown on a silicon wafer. Thus, the grids make possible the concurrent investigation of chemical and structural information on the same sample. Convenient modification of the surfaces of the grids provides access to a wide range of new substrates for the direct imaging of chemically modified surfaces by TEM. We demonstrate the utility of these grids by aligning DNA on the chemically modified SiO2 surface in order to direct the assembly of linear arrays of nanoparticles. Using these grids, we are able to quantify the effects of assembly conditions on nanoparticle size, spacing, and dispersity in the arrays. PMID- 16383341 TI - Purifying barite for oxygen isotope measurement by dissolution and reprecipitation in a chelating solution. AB - In the laboratory, barite precipitated from a solution with a high nitrate/sulfate ratio can have a significant amount (up to 28% by weight) of nitrate occluded in barite crystals that cannot be simply washed away. The impurity poses a serious problem for an accurate measurement of the oxygen isotope compositions for atmospheric sulfate, since atmospheric nitrate bears extremely positive Delta17O and delta18O values. Currently available methods for removing the occluded nitrate are either ineffective or not tested for oxygen isotope exchange. Here, I report a DTPA (a chelating solution) dissolution and reprecipitation (DDARP) method that is simple and effective in removing nitrate and other contaminants in barite. A series of barite dissolution and reprecipitation experiments that utilize 17O-anomalous solutions or barite crystals is conducted to examine the effect on oxygen isotopes during various treatments. It is established that no oxygen isotope exchange occurs between sulfate and water during DDARP treatment at two experimental temperatures (21 and 70 degrees C). Occlusion of DTPA itself in barite is negligible. Upon acidification, barite reprecipitation from a DTPA solution is quantitative (approximately 100%). Partially extracted barite may have slightly lower delta18O or delta34S values than the originals but no effect on Delta17O values. It is also demonstrated that heavily nitrate-contaminated barite samples are free of nitrate occlusion after two dissolution-reprecipitation cycles. PMID- 16383342 TI - Parallel ion parking of protein mixtures. AB - The multiple charging phenomenon resulting from electrospray ionization of proteins, while useful for the ability to make several mass measurements on a single component, can lead to highly complex spectra when mixtures are analyzed, as each component can generate multiple ions of distinct mass-to-charge ratio. Ion/ion proton-transfer reactions can overcome this problem by reduction of all components to the +1 charge state, but this typically requires the ability to extend the mass range of the instrument well beyond that available in most commercial instruments. Furthermore, reduction of protein charge to +1 also results in a reduction in detector response. Here it is shown that application of a relatively high amplitude, low-frequency auxiliary ac signal to the end cap electrodes of a 3-D ion trap during an ion/ion reaction can slow the ion/ion reaction rates of ions over a broad m/z range, in a process termed HALF parallel ion parking. Adjustment of the frequency and amplitude of the applied voltage allows the mass range into which the initial ion signal is moved to be controlled, allowing for the simplification of multicomponent mixtures within a mass range that is more commonly available on commercial systems. In addition to decreasing spectral complexity, this is advantageous for mixtures with low abundance components, as there is less compromise with detector response than in reduction to the +1 charge state. Preliminary evidence also suggests that the ion collision cross section may play an important role in determining which charge states are most significantly inhibited from further ion/ion reactions under a given set of ion parking conditions. PMID- 16383343 TI - Protein recognition via surface molecularly imprinted polymer nanowires. AB - In this paper, we present a technique for the preparation of polymer nanowires with the protein molecule imprinted and binding sites at surface. These surface imprinting nanowires exhibit highly selective recognition for a variety of template proteins, including albumin, hemoglobin, and cytochrome c. This recognition may be through a multistep adsorption, with the specificity conferred by hydrogen bonding and shape selectivity. Due to the protein imprinted sites are located at, or close to, the surface; these imprinted nanowires have a good site accessibility toward the target protein molecules. Furthermore, the large surface area of the nanowires results in large protein molecule binding capacity of the imprinted nanowires. PMID- 16383344 TI - Uncoated, broad fluorescent, and size-homogeneous CdSe quantum dots for bioanalyses. AB - In the present study, we describe the synthesis of highly luminescent uncoated water-soluble CdSe quantum dots (QDs) possessing the following characteristics: approximately 2 nm in diameter, with very good size distribution (in 95% homodispersed) accompanied by a broad-band photoluminescent spectrum. The synthetic procedure is simple, is conducted at room temperature, in the absence of the most popular coordinating ligands (as TOPO or HDA), and is highly reproducible. The obtained CdSe core QDs possessed a comparatively long fluorescence half-life (approximately 30-90 ns, depending on the emission wavelength) detected by time-resolved spectroscopy. These QDs were further conjugated with antibodies and applied in several biochemical analyses. PMID- 16383345 TI - V-type nerve agent detection using a carbon nanotube-based amperometric enzyme electrode. AB - An enzyme electrode for the detection of V-type nerve agents, VX (O-ethyl-S-2 diisopropylaminoethyl methylphosphonothioate) and R-VX (O-isobutyl-S-2 diethylaminoethyl methylphosphonothioate), is proposed. The principle of the new biosensor is based on the enzyme-catalyzed hydrolysis of the nerve agents and amperometric detection of the thiol-containing hydrolysis products at carbon nanotube-modified screen-printed electrodes. Demeton-S was used as a nerve agent mimic. 2-(Diethylamino)ethanethiol (DEAET) and 2-(dimethylamino)ethanethiol (DMAET), the thiol-containing hydrolysis product and hydrolysis product mimic of R-VX and VX, respectively, were monitored by exploiting the electrocatalytic activity of carbon nanotubes (CNT). As low as 2 microM DMAET and 0.8 microM DEAET were detected selectively at a low applied potential of 0.5 V vs Ag/AgCl at a CNT modified mediator-free amperometric electrode. Further, the large surface area and the hydrophobicity of CNT was used to immobilize organophosphorus hydrolase mutant with improved catalytic activity for the hydrolysis of the P-S bond of phosphothiolester neurotoxins including VX and R-VX nerve gases to develop a novel, mediator-free, membrane-free biosensor for V-type nerve agents. The applicability of the biosensor was demonstrated for direct, rapid, and selective detection of V-type nerve agents' mimic demeton-S. The selectivity of the sensor against interferences and application to spiked lake water samples was demonstrated. PMID- 16383346 TI - Firefly bioluminescent assay of ATP in the presence of ATP extractant by using liposomes. AB - Liposomes containing phosphatidylcholine (PC) and cholesterol (Chol) were applied to the enhancer for firefly bioluminescence (BL) assay for ATP in the presence of cationic surfactants using as an extractant for the release of ATP from living cells. Benzalkonium chloride (BAC) was used as an ATP extractant. However, BAC seriously inhibited the activity of luciferase, thus resulting in the remarkable decrease in the sensitivity of the BL assay for ATP. On the other hand, we found that BAC was associated with liposomes to form cationic liposomes containing BAC. The association rate of BAC with liposomes was faster than that of BAC with luciferase. As a result, the inhibitory effect of BAC on luciferase was eliminated in the presence of liposomes. In addition, cationic liposomes thus formed enhanced BL emission. BL measurement conditions were optimized in terms of liposome charge type, liposome size, and total concentration of PC and Chol. ATP can be sensitively determined without dilution of analytical samples by using liposomes. The detection limit of ATP with and without liposomes was 100 amol and 25 fmol in aqueous ATP standard solutions containing 0.06% BAC, respectively. The method was applied to the determination of ATP in Escherichia coli extracts. The BL intensity was linear from 4 x 10(4) to 1 x 10(7) cells mL(-1) in the absence of liposomes. On the other hand, the BL intensity was linear from 4 x 10(3) to 4 x 10(6) cells mL(-1) in the presence of liposomes. The detection limit of ATP in E. coli extracts was improved by a factor of 10 via use of liposomes. PMID- 16383347 TI - A 96-well screen filter plate for high-throughput biological sample preparation and LC-MS/MS analysis. AB - A novel 96-well screen filter plate (patent pending) has been invented to eliminate a time-consuming and labor-intensive step in preparation of in vivo study samples--to remove blood or plasma clots. These clots plug the pipet tips during a manual or automated sample-transfer step causing inaccurate pipetting or total pipetting failure. Traditionally, these blood and plasma clots are removed by picking them out manually one by one from each sample tube before any sample transfer can be made. This has significantly slowed the sample preparation process and has become a bottleneck for automated high-throughput sample preparation using robotic liquid handlers. Our novel screen filter plate was developed to solve this problem. The 96-well screen filter plate consists of 96 stainless steel wire-mesh screen tubes connected to the 96 openings of a top plate so that the screen filter plate can be readily inserted into a 96-well sample storage plate. Upon insertion, the blood and plasma clots are excluded from entering the screen tube while clear sample solutions flow freely into it. In this way, sample transfer can be easily completed by either manual or automated pipetting methods. In this report, three structurally diverse compounds were selected to evaluate and validate the use of the screen filter plate. The plasma samples of these compounds were transferred and processed in the presence and absence of the screen filter plate and then analyzed by LC-MS/MS methods. Our results showed a good agreement between the samples prepared with and without the screen filter plate, demonstrating the utility and efficiency of this novel device for preparation of blood and plasma samples. The device is simple, easy to use, and reusable. It can be employed for sample preparation of other biological fluids that contain floating particulates or aggregates. PMID- 16383348 TI - Electrochemical system for the simultaneous monitoring of algal motility and phototaxis. AB - A compact electrochemical system equipped with dual electrodes was developed for the simultaneous monitoring of motility and phototaxis of flagellates. Photoinduced behavior of unicellular flagellate alga Chlamydomonas reinhardtii in the presence of diltiazem, azide, or ethanol was recorded as changes in the redox currents for a coexisiting redox marker. The system responded to the chemicals in significantly different ways; it allows qualitative and semiquantitative evaluation of influences of chemical species on the algal cells. Therefore, the present system is potentially applicable to not only aquatic risk assessment but also screening of newly found and synthesized compounds in terms of hazard probability and severity. Additionally, it unveiled some new and interesting behaviors of the algae. PMID- 16383349 TI - The science and politics of colorectal cancer screening. PMID- 16383350 TI - New approaches to vaccine adjuvants: inhibiting the inhibitor. PMID- 16383351 TI - Monte Carlo simulation of a model of water. AB - We simulate TIP3P water using a constrained Monte Carlo algorithm to generate electrostatic interactions eliminating the need to sum over long-ranged Coulomb interactions. We study discretization errors when interpolating charges using splines and Gaussians. We compare our implementation to molecular dynamics and Brownian dynamics codes. PMID- 16383352 TI - Self-diffusion of particles in gas-driven granular layers with periodic flow modulation. AB - We study self-diffusion of particles in gas-driven granular layers by high-speed fluorescent video microscopy. We show that periodic flow modulation results in an enhancement of the particle's diffusion. The diffusion enhancement, which in turn is an indication of more efficient fluidization of the granular layer, is associated with the onset of disordered subharmonic patterns. Our measurements provide a sensitive characterization method of the fluidization properties of particulate-gas systems. PMID- 16383353 TI - Electric-field-temperature phase diagram of an antiferroelectric liquid crystal. AB - We present a precise electric-field-temperature phase diagram of an antiferroelectric liquid crystal with a short pitch Sm-Calpha* phase. This was obtained by using a photoelastic modulator. A unique field-induced phase was found inside the Sm-Calpha* phase, which displayed low birefringence. Two tricritical points related to the phase were also observed. In addition, numerical calculations were made based on the discrete phenomenological model. The numerical results reproduced the experimental ones and it was clarified that the phase has a three-layer structure without spatial modulation. PMID- 16383354 TI - Interfacial slip in sheared polymer blends. AB - We have developed a dynamic self-consistent field theory, without any adjustable parameters, for unentangled polymer blends under shear. Our model accounts for the interaction between polymers, and enables one to compute the evolution of the local rheology, microstructure, and the conformations of the polymer chains under shear self-consistently. We use this model to study the interfacial dynamics in sheared polymer blends and make a quantitative comparison between this model and molecular dynamics simulations. We find good agreement between the two methods. PMID- 16383355 TI - Large oxidation dependence observed in terahertz dielectric response for cytochrome c. AB - Far infrared dielectric response is used to characterize the collective mode density of states for cytochrome c as a function of oxidation state and hydration using terahertz time domain spectroscopy. A strong absorbance and refractive index increase was observed with the oxidation. A simple phenomenological fitting using a continuous distribution of oscillators reproduces the frequency dependence of the complex dielectric response as well as demonstrates quantitative agreement with a uniform increase in either mode density or polarizability with oxidation in the 5-80 cm(-1) frequency range. Hydration dependence measurements find that a difference in the equilibrium water content for ferri and ferro cytochrome c is not sufficient to account for the large change in terahertz response. The large dielectric increase at terahertz frequencies with oxidation suggests either a significant global softening of the potential and/or a significant increase in polarizability with oxidation. PMID- 16383356 TI - Strong shock waves and nonequilibrium response in a one-dimensional gas: a Boltzmann equation approach. AB - We investigate the nonequilibrium behavior of a one-dimensional binary fluid on the basis of Boltzmann equation, using an infinitely strong shock wave as probe. Density, velocity, and temperature profiles are obtained as a function of the mixture mass ratio mu. We show that temperature overshoots near the shock layer, and that heavy particles are denser, slower, and cooler than light particles in the strong nonequilibrium region around the shock. The shock width omega(mu), which characterizes the size of this region, decreases as omega(mu) approximately mu(1/3) for mu-->0. In this limit, two very different length scales control the fluid structure, with heavy particles equilibrating much faster than light ones. Hydrodynamic fields relax exponentially toward equilibrium: phi(chi) approximately exp[-chi/lambda]. The scale separation is also apparent here, with two typical scales, lambda1 and lambda2, such that lambda1 approximately mu(1/2 as mu-->0, while lambda2, which is the slow scale controlling the fluid's asymptotic relaxation, increases to a constant value in this limit. These results are discussed in light of recent numerical studies on the nonequilibrium behavior of similar one-dimensional binary fluids. PMID- 16383357 TI - First passage time of N excluded-volume particles on a line. AB - Motivated by recent single-molecule studies of proteins sliding on a DNA molecule, we explore the targeting dynamics of particles ("proteins") sliding diffusively along a line ("DNA") in search of their target site (specific target sequence). At lower particle densities, one observes an expected reduction of the mean first passage time proportional to N(-2), with corrections at higher concentrations. We explicitly take adsorption and desorption effects, to and from the DNA, into account. For this general case, we also consider finite-size effects when the continuum approximation based on the number density of particles breaks down. Moreover, we address the first-passage-time problem of a tagged particle diffusing among other particles. PMID- 16383358 TI - Similarity of fluctuations in correlated systems: the case of seismicity. AB - We report a similarity of fluctuations in equilibrium critical phenomena and nonequilibrium systems, which is based on the concept of natural time. The worldwide seismicity as well as that of the San Andreas fault system and Japan are analyzed. An order parameter is chosen and its fluctuations relative to the standard deviation of the distribution are studied. We find that the scaled distributions fall on the same curve, which interestingly exhibits, over four orders of magnitude, features similar to those in several equilibrium critical phenomena (e.g., two-dimensional Ising model) as well as in nonequilibrium systems (e.g., three-dimensional turbulent flow). PMID- 16383359 TI - Acceleration of diffusion in randomly switching potential with supersymmetry. AB - We investigate the overdamped Brownian motion in a supersymmetric periodic potential switched by Markovian dichotomous noise between two configurations. The two configurations differ from each other by a shift of one-half period. The calculation of the effective diffusion coefficient is reduced to the mean first passage time problem. We derive general equations to calculate the effective diffusion coefficient of Brownian particles moving in arbitrary supersymmetric potential. For the sawtooth potential, we obtain the exact expression for the effective diffusion coefficient, which is valid for the arbitrary mean rate of potential switchings and arbitrary intensity of white Gaussian noise. We find the acceleration of diffusion in comparison with the free diffusion case and a finite net diffusion in the absence of thermal noise. Such a potential could be used to enhance the diffusion over its free value by an appropriate choice of parameters. PMID- 16383360 TI - Magnetic properties in monolayers of a model polydisperse ferrofluid. AB - The influence of polydispersity on the equilibrium properties of monolayers of three-dimensional dipolar spheres with short-range repulsive interactions is studied by means of Monte Carlo simulations and a high field approximation perturbation theory. The particle distribution in the simulations is realized in the semigrand ensemble by tuning appropriately the underlying particle distribution density. The magnetization curves are calculated as functions of density and temperature, and the obtained results are compared with the data determined in a monodisperse equivalent of the system. In-plane and out-of-plane initial magnetic susceptibilities are determined using external fields applied parallel or normal to the monolayer plane. Susceptibility data for the true two- and three-dimensional counterparts of the system are also calculated for comparison. Our findings for the magnetic properties can partly be explained by the structural characteristics obtained from the simulations. PMID- 16383361 TI - Heterogeneity and growing length scales in the dynamics of kinetically constrained lattice gases in two dimensions. AB - We study dynamical heterogeneity and growing dynamical length scales in two kinetically constrained models, namely, the one- and two-vacancy assisted triangular lattice gases. One of the models is a strong glassformer and the other is a fragile glassformer. Both exhibit heterogeneous dynamics with broadly distributed time scales as seen in the distribution of persistence times. We show that the Stokes-Einstein relation is violated, to a greater degree in the fragile glassformer, and show how this violation is related to dynamic heterogeneity. We extract dynamical length scales from structure factors of mobile particles and show, quantitatively, the growth of this length scale as density increases. We comment on how the scaling of lengths and times in these models relates to that in facilitated spin models of glasses. PMID- 16383362 TI - Diffusion of water in confined geometry: the case of a multilamellar bilayer. AB - The diffusion of water confined in a stack of GM3 ganglioside bilayers is studied by computer simulation. A theoretical analysis of the behavior of the mean square displacement parallel and perpendicular to the bilayer surface is also provided in terms of diffusion equations in a very long time interval, between 2 ps and 2 ns. Such an analysis has allowed us to identify two different time regimes, to clarify the nature of nonlinear time dependence of the mean square displacement, and to give an interpretation to the origin of the often used time dependent diffusion coefficient. Both the effects of spatial inhomogeneities and boundary conditions are demonstrated to be the key points for the interpretation of all these results within a consistent theoretical framework. PMID- 16383363 TI - Molecular thermodiffusion (thermophoresis) in liquid mixtures. AB - Thermodiffusion (thermophoresis) in liquid mixtures is theoretically examined using a hydrodynamic approach. Thermodiffusion is related to the local temperature-induced pressure gradient in the liquid layer surrounding the selected molecule and to the secondary macroscopic pressure gradient established in the system. The local pressure gradient is produced by excess pressure due to the asymmetry of interactions with surrounding molecules in a nonuniform temperature field. The secondary pressure gradient is considered an independent parameter related to the concentration gradient formed by volume forces, calculated from the generalized equations for mass transfer. Values of Soret coefficients for mixtures of toluene and -hexane are calculated using parameters in the literature. When the molecules are assumed to be similar in shape, the calculated Soret coefficients are lower than the empirical values found in the literature. However, by introducing an asymmetry parameter, which is calculated from independent measurements of component diffusion in the literature, very good agreement is obtained. PMID- 16383364 TI - Simplified thermodynamic model for equilibrium capillary pressure in a fractal porous medium. AB - Defining a relation for equilibrium pressure in a porous medium has been difficult to do in terms of readily measurable parameters. We present a simplified analysis of this problem using the first law of thermodynamics combined with a fractal description of a porous system. The results show that the variation in fluid interfacial area with fluid volume, and the respective interfacial surface tensions, are dominant factors determining equilibrium capillary pressure. Departures from equilibrium are seen to occur when fluid solid contact lines are in movement. By describing the pore space as fractal we are able to obtain an expression for the change in fluid interfacial area with respect to its volume, and the resulting model shows a strong fit to pressure data obtained from a capillary rise experiment conducted in a coarse-grained SiO2 sand. PMID- 16383365 TI - Power-law exponents for the shear viscosity of non-Newtonian simple fluids. AB - Nonequilibrium molecular-dynamics simulations are performed to compute the shear viscosities of a simple Lennard-Jones fluid across a wide range of densities and temperatures that span the liquid phase. It is found that the standard mode coupling value of beta = 0.05 for the exponent of the strain rate power-law dependence (eta = eta0-eta1gammabeta) is only applicable in a very narrow region of the thermodynamic state-space. More generally, the exponent is a remarkably simple linear function of temperature and density, analogous to the linear relationship that exists for the scaling exponents of the pressure and energy found previously by Ge et al. [Phys. Rev. E 67, 061201 (2003)], and ranges between approximately 0.2 and 1.6. It is also found that the parameters eta0 and eta1 are steep functions of increasing density for any particular temperature and can be represented by a stretched exponential of the density. PMID- 16383366 TI - Mathematical analysis of thermal diffusion shock waves. AB - Thermal diffusion, also known as the Ludwig-Soret effect, refers to the separation of mixtures in a temperature gradient. For a binary mixture the time dependence of the change in concentration of each species is governed by a nonlinear partial differential equation in space and time. Here, an exact solution of the Ludwig-Soret equation without mass diffusion for a sinusoidal temperature field is given. The solution shows that counterpropagating shock waves are produced which slow and eventually come to a halt. Expressions are found for the shock time for two limiting values of the starting density fraction. The effects of diffusion on the development of the concentration profile in time and space are found by numerical integration of the nonlinear differential equation. PMID- 16383367 TI - Collapses of two-dimensional granular columns. AB - The first detailed quantitative observations of the two-dimensional collapse of a granular column along a horizontal channel are presented for a variety of materials. Together with the complementary study for the axisymmetric situation, we conclude that for granular collapses the generally accepted approaches, that are highly dependent on frictional parameters, do not describe the main flow phenomena. The motion divides in two main flow regimes at a approximately 1.8, where the aspect ratio a = hi/di and hi and di are the initial height and width of the column. We describe the details of collapse by emphasizing the sequential occurrence of a main spreading followed by a final avalanching phase. For the low a regime, a < 1.8, we derive descriptions of the final geometry by direct physical arguments. For the large a regime, a > 1.8, we determine that nearly all details of the collapse, including the position of the flow front as a function of time, the emplacement time, the self-similar final profiles, and especially their maximum vertical and horizontal extension, are established during the spreading phase and can be expressed in terms of the initial geometrical parameters but are independent of basal and internal friction parameters. PMID- 16383368 TI - Characterization of the material response in granular ratcheting. AB - The existence of a very special ratcheting regime has recently been reported in a granular packing subjected to cyclic loading. In this state, the system accumulates a small permanent deformation after each cycle. The value of this permanent strain accumulation becomes independent of the number of cycles after a short transient regime. We show in this paper that a characterization of the material response in this peculiar state is possible in terms of three simple macroscopic variables. The definition of these variables is such that they can be easily measured both in the experiments and in the simulations. A thorough investigation of the micro- and macromechanical factors affecting these variables has been carried out by means of molecular-dynamics simulations of a polydisperse disk packing, as a simple model system for granular material. Biaxial test boundary conditions with periodically varying load were implemented. The effect on the plastic response of the confining pressure, the deviatoric stress, and the number of cycles has been investigated. The stiffness of the contacts and friction has been shown to play an important role in the overall response of the system. Especially illustrative is the influence of the peculiar hysteretical behavior in the stress-strain space on the accumulation of permanent strain and the energy dissipation. PMID- 16383369 TI - Local heat flux and energy loss in a two-dimensional vibrated granular gas. AB - We performed event-driven simulations of a two-dimensional granular gas between two vibrating walls and directly measured the local heat flux and local energy dissipation in the stationary state. Describing the local heat flux as a function of the coordinate in the direction perpendicular to the driving walls, we test a generalization of Fourier's law, q(x)=-kappa inverted delta T(x)+mu inverted delta rho(x), by relating the local heat flux to the local gradients of the temperature and density. This ansatz accounts for the fact that heat flux can also be generated by density gradients, not only by temperature gradients. Assuming the transport coefficients kappa and mu to be independent of x, we check the validity of this assumption and test the generalized Fourier law in the simulations. Both kappa and mu are determined for different system parameters, in particular, for a wide range of coefficients of restitution. We also compare our numerical results to existing hydrodynamic theories. Agreement is found for kappa for very small inelasticities only, i.e., when the gradients are small. Beyond this region, kappa and mu exhibit a striking nonmonotonic behavior. This may hint that hydrodynamics to Navier-Stokes order cannot be applied to moderately inelastic vibrated systems. PMID- 16383370 TI - Impulse absorption by tapered horizontal alignments of elastic spheres. AB - We present an analytical and numerical study of the problem of mechanical impulse propagation through a horizontal alignment of progressively shrinking (tapered) elastic spheres that are placed between two rigid end walls. The studies are confined to cases where initial loading between the spheres is zero (i.e., in the "sonic vacuum" region). The spheres are assumed to interact via the Hertz potential. Force and energy as a function of time for selected grains that comprise the solitary wave are provided and shed light on the system's behavior. Propagation of energy is analytically studied in the hard-sphere approximation and phase diagrams plotting normalized kinetic energy of the smallest grain at the tapered end are developed for various chain lengths and tapering factors. These details are then compared to kinetic energy phase diagrams obtained via extensive dynamical simulations. Our figures indicate that the ratios of the kinetic energies of the smallest to largest grains possess a Gaussian dependence on tapering and an exponential decay when the number of grains increases. The conclusions are independent of system size, thus being applicable to tapered alignments of micron-sized spheres as well as those that are macroscopic and more easily realizable in the laboratory. Results demonstrate the capabililty of these chains to thermalize propagating impulses and thereby act as potential shock absorbing devices. PMID- 16383371 TI - Dynamics of shallow impact cratering. AB - We present data for the time dependence of wooden spheres penetrating into a loose noncohesive packing of glass beads. The stopping time is a factor of 3 longer than the time d/v0 needed to travel the total penetration distance d at the impact speed v0. The acceleration decreases monotonically throughout the impact. These kinematics are modeled by a position- and velocity-dependent stopping force that is constrained to reproduce prior observations for the scaling of the penetration depth with the total drop distance. PMID- 16383372 TI - Influence of image resolution and evaluation algorithm on estimates of the lacunarity of porous media. AB - In recent years, experience has demonstrated that the classical fractal dimensions are not sufficient to describe uniquely the interstitial geometry of porous media. At least one additional index or dimension is necessary. Lacunarity, a measure of the degree to which a data set is translationally invariant, is a possible candidate. Unfortunately, several approaches exist to evaluate it on the basis of binary images of the object under study, and it is unclear to what extent the lacunarity estimates that these methods produce are dependent on the resolution of the images used. In the present work, the gliding box algorithm of Allain and Cloitre [Phys. Rev. A 44, 3552 (1991)] and two variants of the sandbox algorithm of Chappard et al. [J. Pathol. 195, 515 (2001)], along with three additional algorithms, are used to evaluate the lacunarity of images of a textbook fractal, the Sierpinski carpet, of scanning electron micrographs of a thin section of a European soil, and of light transmission photographs of a Togolese soil. The results suggest that lacunarity estimates, as well as the ranking of the three tested systems according to their lacunarity, are affected strongly by the algorithm used, by the resolution of the images to which these algorithms are applied, and, at least for three of the algorithms (producing scale-dependent lacunarity estimates), by the scale at which the images are observed. Depending on the conditions under which the estimation of the lacunarity is carried out, lacunarity values range from 1.02 to 2.14 for the three systems tested, and all three of the systems used can be viewed alternatively as the most or the least "lacunar." Some of this indeterminacy and dependence on image resolution is alleviated in the averaged lacunarity estimates yielded by Chappard et al.'s algorithm. Further research will be needed to determine if these lacunarity estimates allow an improved, unique characterization of porous media. PMID- 16383373 TI - Characterization of force chains in granular material. AB - It has been observed that the majority of particles in a granular material carries less than the average load and that the number of particles carrying larger than the average load decreases exponentially with increasing contact force. The particles carrying above average load appear to form a strong network of forces while the majority of particles belong to a weak network. The strong network of forces appear to have a spatial characteristic whereby the stronger forces are carried though chainlike particle groups referred to as force chains. There is a strong case for a connection between force chains of the discrete medium and the trajectory of the most compressive principal stress in its continuous idealization. While such properties seem obvious from descriptive analysis of physical and numerical experiments in granular media, progress in quantification of the force chain statistics requires an objective description of what constitutes a force chain. A procedure to quantify the occurrence of force chains is built on a proposed definition having two parts: first, the chain is a quasilinear arrangement of three or more particles, and second, along the chain, stress concentration within each grain is characterized by the vector delineating the most compressive principal stress. The procedure is incorporated into an algorithm that can be applied to large particle assemblies to compile force chain statistics. The procedure is demonstrated on a discrete element simulation of a rigid punch into a half space. It was found that only approximately half of the particles within the group of so-called strong network particles are part of force chains. Throughout deformation, the average length of force chains varied slightly but the number of force chains decreased as the punch advanced. The force chain lengths follow an exponential distribution. The procedure provides a tool for objective analysis of force chains, although future work is required to incorporate branching of force chains into the analysis. PMID- 16383374 TI - Friction versus texture at the approach of a granular avalanche. AB - We perform an analysis of the granular texture of a granular bed close to stability limit. Our analysis is based on a unique criterion of friction mobilization in a simulated two-dimensional packing. In this way, we recover the bimodal character of granular texture and the coexistence of weak and strong phases in the sense of distinct contacts populations. Moreover, we show the existence of a well-defined subset of contacts within the weak contact network. These contacts are characterized by their important friction and form a highly coherent population in terms of fabric. They play an antagonistic role with respect to force chains. Thus, we are able to discriminate between incoherent contacts and coherent contacts in the weak phase and to specify the role that the latter plays in the destabilization process. PMID- 16383375 TI - Microstructure evolution of hydrated cement pastes. AB - We propose an original method based on both proton nuclear magnetic relaxation dispersion and high-resolution NMR spectra to investigate the microstructure of synthesized Ca3SiO5-hydrated cement paste. This method allows a clear assessment of the local proton chemical sites as well as the determination of dynamical information of moving proton species in pores. We show also how the microstructure evolves during and after completion of hydration in a range of length scales between 2 and 500 nm. In particular, we show how the pore size distribution of the cement paste reaches progressively a power-law characteristic of a surface-fractal distribution with a dimension Df = 2.6, which takes into account the hierarchical order in the material. Last, we study how this pore size distribution is modified during setting by varying either the water-to-cement ratio or addition of ultrafine particles. This shows that our method could be relevant to relate the mechanical properties to the microstructure of the material. This proposed NMR method is general enough for the characterization of microstructure of any porous media with reactive surface involving water confinement. PMID- 16383376 TI - Quenched charge disorder and Coulomb interactions. AB - We develop a general formalism to investigate the effect of quenched fixed charge disorder on effective electrostatic interactions between charged surfaces in a one-component (counterion-only) Coulomb fluid. Analytical results are explicitly derived for two asymptotic and complementary cases: (i) mean-field or Poisson Boltzmann limit (including Gaussian-fluctuations correction), which is valid for small electrostatic coupling, and (ii) strong-coupling limit, where electrostatic correlations mediated by counterions become significantly large as, for instance, realized in systems with high-valency counterions. In the particular case of two apposed and ideally polarizable planar surfaces with equal mean surface charge, we find that the effect of the disorder is nil on the mean-field level and thus the plates repel. In the strong-coupling limit, however, the effect of charge disorder turns out to be additive in the free energy and leads to an enhanced long-range attraction between the two surfaces. We show that the equilibrium interplate distance between the surfaces decreases for elevated disorder strength (i.e., for increasing mean-square deviation around the mean surface charge), and eventually tends to zero, suggesting a disorder-driven collapse transition. PMID- 16383377 TI - Polarized small-angle light scattering from gels estimated in terms of a statistical approach. AB - To analyze polarized light scattering patterns from gels, an approach is proposed to calculate the scattered intensity. In the proposed model system, difference between polar angles of the principal axes of the and elements, which were defined with respect to the axis along the distance between two elements, was given as a correlation of the distance between the two elements. Furthermore, the azimuthal angle, which makes a projection of the principal axis onto a plane perpendicular to the principal axis of the element, was also given as a correlation of the distance between the two elements. The theoretical calculation was carried out for the scattered intensity under Hv and Vv polarization conditions. The general equations proposed for Hv and Vv scattering were based on a statistical approach for polarized light scattering system. The calculated pattern under the Hv polarization condition showed an X-type pattern and was in good agreement with the pattern observed from polymer gels prepared by quenching their solutions to the desired temperatures. PMID- 16383378 TI - Invasion percolation between two sites. AB - We investigate the process of invasion percolation between two sites (injection and extraction sites) separated by a distance r in two-dimensional lattices of size L. Our results for the nontrapping invasion percolation model indicate that the statistics of the mass of invaded clusters is significantly dependent on the local occupation probability (pressure) Pe at the extraction site. For Pe = 0, we show that the mass distribution of invaded clusters P(M) follows a power-law P(M) approximately M(-alpha) for intermediate values of the mass M, with an exponent alpha = 1.39+/-0.03. When the local pressure is set to Pe = Pc, where Pc corresponds to the site percolation threshold of the lattice topology, the distribution P(M) still displays a scaling region, but with an exponent alpha = 1.02+/-0.03. This last behavior is consistent with previous results for the cluster statistics in standard percolation. In spite of these differences, the results of our simulations indicate that the fractal dimension of the invaded cluster does not depend significantly on the local pressure Pe and it is consistent with the fractal dimension values reported for standard invasion percolation. Finally, we perform extensive numerical simulations to determine the effect of the lattice borders on the statistics of the invaded clusters and also to characterize the self-organized critical behavior of the invasion percolation process. PMID- 16383379 TI - Criticality in charge-asymmetric hard-sphere ionic fluids. AB - Phase separation and criticality are analyzed in z:1 charge-asymmetric ionic fluids of equisized hard spheres by generalizing the Debye-Huckel approach combined with ionic association, cluster solvation by charged ions, and hard-core interactions, following lines developed by Fisher and Levin for the 1:1 case (i.e., the restricted primitive model). Explicit analytical calculations for 2:1 and 3:1 systems account for ionic association into dimers, trimers, and tetramers and subsequent multipolar cluster solvation. The reduced critical temperatures, Tc* (normalized by z), decrease with charge asymmetry, while the critical densities increase rapidly with . The results compare favorably with simulations and represent a distinct improvement over all current theories such as the mean spherical approximation, symmetric Poisson-Boltzmann theory, etc. For z not equal to 1, the interphase Galvani (or absolute electrostatic) potential difference, Deltaphi(T), between coexisting liquid and vapor phases is calculated and found to vanish as absolute value (T-Tc) beta when T-->Tc-with, since our approximations are classical, beta = (1/2). Above Tc, the compressibility maxima and so-called k-inflection loci (which aid the fast and accurate determination of the critical parameters) are found to exhibit a strong z dependence. PMID- 16383380 TI - Melting and evaporation in classical two-dimensional clusters confined by a Coulomb potential. AB - The thermal properties of a two-dimensional classical cluster of negatively charged particles bound by a punctual positive charge are presented. The melting phenomenon is analyzed and the features which characterize such a solid-liquid transition are highlighted. We found that the presence of metastable states strongly modifies the melting scenario, and that the melting temperature of the system is determined by the height of the saddle point energy separating the ground state and the metastable state. Due to the particular type of confinement potential considered in this paper, we also found that, at sufficiently large temperature, the cluster can become thermally ionized. PMID- 16383381 TI - Entropic attraction and fluid-glass transition in micellar solutions of associative diblock copolymers. AB - We report quantitative determination of the strength of attraction between spherical micelles of associative diblock copolymers. We offer detailed characterization of the dilute micellar solutions by neutron and dynamic light scattering techniques and by viscometry, and we interpret the data with the aid of the adhesive hard-sphere model. This model permits estimate of the stickiness parameter for varying fraction of stickers on the micelles. At this range of attraction the solutions exhibit, in the crowded regime, rheological and dynamics behavior representative of both repulsive caging and attractive bonding glassy dynamics. PMID- 16383382 TI - Stabilization of nonlinear velocity profiles in athermal systems undergoing planar shear flow. AB - We perform molecular dynamics simulations of model frictionless granular systems undergoing boundary-driven planar shear flow in two spatial dimensions with the goal of developing a more complete understanding of how dense particulate systems respond to applied shear. In particular, we are interested in determining when these systems will possess linear velocity profiles and when they will develop highly localized velocity profiles in response to shear. In previous work on similar systems we showed that nonlinear velocity profiles form when the speed of the shearing boundary exceeds the speed of shear waves in the material. However, we find that nonlinear velocity profiles in these systems are unstable at very long times. The degree of nonlinearity slowly decreases in time; the velocity profiles become linear when the granular temperature and density profiles are uniform across the system at long times. We measure the time t(l) required for the velocity profiles to become linear and find that t(l) increases as a power law with the speed of the shearing boundary and increases rapidly as the packing fraction approaches random close packing. We also performed simulations in which differences in the granular temperature across the system were maintained by vertically vibrating one of the boundaries during shear flow. We find that nonlinear velocity profiles form and are stable at long times if the difference in the granular temperature across the system exceeds a threshold value that is comparable to the glass transition temperature in an equilibrium system at the same average density. Finally, the sheared and vibrated systems form stable shear bands, or highly localized velocity profiles, when the applied shear stress is lowered below the yield stress of the static part of the system. PMID- 16383383 TI - Pressure dependence of the glass temperature in supercooled liquids. AB - The description of the pressure evolution of the glass temperature Tg(p) based on experimental data for diethyl phtalate is discussed. First, parameterizations of Tg(P) experimental data applied are briefly given. Then a novel relation based on the modified Simon-Glatzel equation is proposed. Its applications may result in the appearance of the asymptotic temperature (theta) and the asymptotic pressure (pi) previously postulated [E. Donth,, Springer Series in Material Sci. II (Springer, Berlin, 1998), Vol. 48, pp. 6, 375]. The asymptotic pressure is hidden in the negative pressure domain. Such asymptotic behavior was absent for parameterizations of Tg(p) data in glassy liquids applied up to now. PMID- 16383384 TI - Melting of quasi-two-dimensional crystalline Pb supported on liquid Ga. AB - Experimental studies have shown that the Pb monolayer that segregates in the PbGa alloy liquid-vapor interface forms a two-dimensional hexagonal crystal that melts at 341 K, and it has been speculated that the disordered phase formed is hexatic. This paper reports the results of simulation studies of the in-plane structure of the outermost stratum of the liquid-vapor interface of a dilute Pb in Ga alloy. These simulations are based on four major improvements to a previous study. First, the simulation studies involve considerably more atoms and considerably longer equilibration runs than considered in the previous work of Chekmarev, Oxtoby, and Rice. Second, a more accurate nonlocal pseudopotential representation of the interactions in the system is used. Third, the amplitude of the out-of plane motion of the Pb atoms is constructed to have the observed value. Fourth, an approximation to the role of the liquid Ga substrate is provided by adding a layer of Ga atoms to the layer of Pb atoms. The results of our simulation studies show that the Ga layer adjacent to the Pb layer has a profound influence on that layer's properties. In particular, it is shown that in the two-layer PbGa system the Pb layer forms, at low temperature, a stable two-dimensional crystal on top of liquid Ga. This two-dimensional crystal melts at a temperature close to that found experimentally. It is found that the crystalline Pb layer is transformed to the liquid state via two intermediate hexatic phases that differ in the magnitude of the bond orientation order. Each of the phase transitions along this melting pathway is first order. The temperature range over which each hexatic phase is stable is small. The profound influence of out-of-plane motion is demonstrated by a comparison of the results of simulations of a quasi-two-dimensional (Q2D) and of a strictly two-dimensional monolayer of Pb. The melting transition in the Q2D one-layer system is first order, directly to the liquid, with no intervention of a hexatic phase. The melting transition in the strictly 2D system involves two stages: a first-order transition to an intermediate hexatic phase followed by melting of the hexatic to a liquid phase. The latter transition is continuous over a small temperature range. An examination of the role of defects in the melting process reveals a picture rather different from that postulated in the Kosterlitz-Thouless-Halperin-Nelson-Young theory of 2D melting. PMID- 16383385 TI - Heterogeneities in the glassy state. AB - We study heterogeneities in a binary Lennard-Jones system below the glass transition using molecular dynamics simulations. We identify mobile and immobile particles and measure their distribution of vibrational amplitudes. For temperatures near the glass transition the distribution of vibrational amplitudes obeys scaling and compares reasonably well with a mean-field theory for the amorphous solid state. To investigate correlations among the immobile and mobile particles we identify clusters and analyze their size and shape. For a fixed number of immobile particles we observe that the immobile particles cluster more strongly together as the temperature is increased which allows the particles to block each other more effectively and to therefore stay immobile. For the mobile particles, on the other hand, the clustering is most pronounced at low temperatures, indicating that mobility at low temperatures can only be sustained in cooperative motion. PMID- 16383386 TI - Interactions between a rotating polarized sphere and a stationary one in an electric field. AB - Precise measurement of the attracting force between two polarized spheres inside an electric field indicates that the rotation of one sphere along the axis perpendicular to the electric field reduces the attracting force between them. The important difference between the experimental results and the existing theory indicated that this reduction is due to several factors. In addition to the reduction of polarization due to the free surface charges, the rotation may also weaken the local field near the rotating sphere, making the main contribution to the reduction of the attracting force. Moreover, the experiment also suggests that the polarization due to the molecular polarizability cannot be ignored. PMID- 16383387 TI - Simple tools for complex phenomena: viscoelastic phase separation captured by disconnectable springs. AB - Viscoelastic phase separation is characterized by the formation of a transient gel upon phase separation. A transient gel state is widely observed in complex fluids including polymer solutions, colloidal suspensions, and protein solutions, but its physical description is quite difficult due to its intrinsically nonequilibrium nature. We have modeled this transient gel state using a type of Brownian dynamics simulation in which coarse-grained particles interacting with a Lennard-Jones potential are connected by elastic springs, which can be disconnected with the probability controlled by the ratio of the stored elastic energy to the thermal energy. The simulations well reproduce pattern evolution in a transient gel of a polymer solution. Our simulations indicate that domain morphology is controlled by two key physical factors: (i) the ratio between the nucleation and growth rates of domains of the less viscoelastic phase and (ii) the fragility of the transient gel. PMID- 16383388 TI - Active microrheology of networks composed of semiflexible polymers: computer simulation of magnetic tweezers. AB - We have simulated the motion of a bead subjected to a constant force while embedded in a network of semiflexible polymers which can represent actin filaments. We find that the bead displacement obeys the power law x approximately t(alpha). After the initial stage characterized by the exponent alpha1 approximately 0.75, we find a different regime with alpha2 approximately 0.5. The response in this regime is linear in force and scales with the polymer concentration as c(-1.4). We find that the polymers pile up ahead of the moving bead, while behind it the polymer density is reduced. We show that the force resisting the bead motion is due to steric repulsion exerted by the polymers on the front hemisphere of the bead. PMID- 16383389 TI - Active microrheology of networks composed of semiflexible polymers: theory and comparison with simulations. AB - Based on the results of our computer simulation [Ter-Oganessian et al., Phys. Rev. E 72, 041510 (2005)], we have developed a theoretical description of the motion of a bead, embedded in a network of semiflexible polymers (filaments) and responding to an applied force. The theory reveals the existence of an osmotic restoring force, generated by the piling up of filaments in front of the moving bead and first deduced through computer simulations. The theory predicts that the bead displacement scales like x approximately t(alpha) with time, where alpha = (1/2) in an intermediate-time regime and alpha = 1 in a long-time regime. It also predicts that the compliance varies with a concentration like c(4/3) in agreement with experiment. PMID- 16383390 TI - Critical adsorption at silicon surfaces in binary liquid mixtures. AB - In critical binary liquid mixtures the preferential adsorption that occurs at liquid-vapor or liquid-solid surfaces is expected to be described by a universal surface scaling function. In this paper, we show that aniline strongly adsorbs at an oxide-coated Si wafer surface from a critical mixture of aniline + cyclohexane where this solid-liquid adsorption can be described by the same universal function found at liquid-vapor surfaces. For a tetrabromoethane + n-dodecane critical mixture the n-dodecane adsorption on an alkylsilane coated Si wafer cannot be described by previously determined adsorption functions. We speculate that this discrepancy is caused by chemical heterogeneities at the alkylsilane surface due to differing surface distributions of -CH3 and -CH2- groups within the silane layer. PMID- 16383391 TI - Capillary condensation in cylindrical nanopores. AB - Using grand canonical Monte Carlo simulations, we have explored the phenomenon of capillary condensation (CC) of Ar at the triple temperature inside infinitely long, cylindrical pores. Pores of radius R = 1 nm, 1.7 nm, and 2.5 nm have been investigated using a gas-surface interaction potential parametrized by the well depth D of the gas on a planar surface made of the same material as that comprising the porous host. For strongly attractive situations--i.e., large D- one or more (depending on R) Ar layers adsorb successively before liquid fills the pore. For very small values of D, in contrast, negligible adsorption occurs at any pressure P below saturated vapor pressure P0; above saturation, there eventually occurs a threshold value of P at which the coverage jumps from empty to full, nearly discontinuously. Hysteresis is found to occur in the simulation data whenever abrupt CC occurs--i.e., for R > or = 1.7 nm--and for small D when R = 1 nm. Then, the pore-emptying branch of the adsorption isotherm exhibits larger coverage than the pore-filling branch, as is known from many experiments and simulation studies. The relation between CC and wetting on planar surfaces is discussed in terms of a threshold value of D, which is about one-half of the value found for the wetting threshold on a planar surface. This finding is consistent with a simple thermodynamic model of the wetting transition developed previously. PMID- 16383392 TI - Chemosensitive running droplet. AB - Chemical control of the spontaneous motion of a reactive oil droplet moving on a glass substrate under an aqueous phase is reported. Experimental results show that the self-motion of an oil droplet is confined on an acid-treated glass surface. The transient behavior of oil-droplet motion is also observed with a high-speed video camera. A mathematical model that incorporates the effect of the glass surface charge is built based on the experimental observation of oil droplet motion. A numerical simulation of this mathematical model reproduced the essential features concerning confinement of oil droplet motion within a certain chemical territory and also its transient behavior. Our results may shed light on physical aspects of reactive spreading and a chemotaxis in living things. PMID- 16383393 TI - Simulation of colloidal crystallization on finite structured templates. AB - We present a numerical study of colloidal crystal growth on finite templates. Specifically, we consider planar, crystalline templates with the structure of the 100, 110, and 100 faces of a fcc crystal. We explore how the size of the induced crystallites depends on template area, lattice spacing and degree of supersaturation. We find that thermal fluctuations of the templating particles around their average positions have a strong effect on the size of the crystallites that grow epitaxially. If the fluctuations exceed the Lindemann criterion, the templates cease to function as a crystallization seed. We find that our numerical results are well described by a suitably modified version of classical nucleation theory. PMID- 16383394 TI - Tetrahedral calcite crystals facilitate self-assembly at the air-water interface. AB - Calcite crystals often nucleate and grow in solutions of calcium carbonate, and these crystallites can become trapped at the air water interface, where they form unusual structures. The most common is a fractal structure, which can extend over a large fraction of the interface, and whose origin is understood in terms of the aggregation of the particles. Much more rarely, a different and entirely unexpected structure is observed: the particles remain well separated on the interface, forming an ordered phase reminiscent of a two-dimensional colloidal crystal. The structure of the crystallites that form this ordered phase is always observed to be tetrahedral, in contrast to the much more common rhombohedral structure of the crystallites that form the fractal phase. We show that the interparticle interaction potential that leads to this ordered phase is a balance between a long-range attractive interaction and a long-range repulsive interaction. The attraction results from gravity-induced capillary forces, while the repulsion results from a dipole-dipole interaction due to the charged surface of the tetrahedral crystals. The interaction potential is estimated from the thermal motion of the particles, and fits to the theoretically expected values suggest that the effective surface charge on the tetrahedral crystals is sigma approximately 0.01 charges/nm2. PMID- 16383395 TI - Neutron reflectivity studies of critical adsorption: behavior of the surface scaling function. AB - Neutron reflectometry has been employed to examine the nature of the critical adsorption surface scaling function for a near-critical mixture of hexane d14+perfluorohexane adsorbing to a solid substrate from the liquid one-phase region. The analysis method of Dietrich and Schack has been applied to examine the nature of the power-law part of the critical adsorption surface scaling function, which has been found to behave as m(z) approximately P0z(-mu) as the critical point is approached. Values of mu = 0.514+/-0.018 and P0 = 0.90+/-0.04 have been obtained. These values are consistent with theoretical expectations (mu(th) = 0.516+/-0.004; P0(th) = 0.94+/-0.05), the value determined from Monte Carlo simulations (P(MC)0 = 0.866, and other experimental determinations (P(ex)0 = 0.955=/-0.08). PMID- 16383396 TI - Determination of piezoelectric and flexoelectric polarization in ferroelectric liquid crystals. AB - A method of determining the local polarization in chiral smectic liquid crystals is proposed. The method consists of simultaneous measurement of the electric permittivity and depth of electro-optic modulation in a helical liquid crystal using weak ac electric field. The spontaneous polarization of a single smectic layer determined in this way contains both the flexoelectric and piezoelectric component. On the other hand, the polarization measured using strong electric fields in a switching experiment contains the piezoelectric component only. The comparison of polarization obtained using strong and weak fields makes it possible to determine the value of flexoelectric polarization. This kind of measurement has been performed for two ferroelectric liquid crystals having quite small absolute values of the spontaneous polarization (few nC/cm2). It turned out that in both cases, the flexoelectric polarization is of the same order of magnitude as the piezoelectric one. PMID- 16383397 TI - Weakly first-order character of the nematic-isotropic phase transition in liquid crystals. AB - The classification of phase transitions in first-order and second-order (or continuous) ones is widely used. The nematic-to-isotropic (NI) transition in liquid crystals is a weakly first-order transition, with only small discontinuities in enthalpy and specific volume at the transition which are not always easy to measure. On the other hand, fluctuation effects near the transition, typical for a continuous transition, are present because of the only weakly first-order character. In a recent paper [Phys. Rev. E 69, 022701 (2004)], it was concluded from the static dielectric permittivity in the isotropic phase near the NI transition that less polar mesogens (with little or no pretransitional effects) are characteristic for a first-order NI phase transition, whereas in the case of strongly polar ones (with large pretransitional effects) the NI transition is close to second order. In this paper, we address the question whether it is, indeed, possible to use these fluctuation effects in the isotropic phase to quantify the "strength" of a weakly first-order transition, i.e., how far it is from second order. Therefore, we measured the temperature dependence of the enthalpy near the NI transition of seven liquid crystals with adiabatic scanning calorimetry and compared the measured values of the latent heat with pretransitional effects in the dielectric constant and the specific heat capacity. The compounds used in the comparison are MBBA, 5CB, 8CB, 5NCS, 5CN, 8CHBT, and D7AB. From our analysis we find, contrary to the assertion in the above reference, no correlation between the strength of the NI transition of a given compound and the pretransitional effects observed, neither dielectrically, nor thermally. PMID- 16383398 TI - Maier-Saupe model of liquid crystals: isotropic-nematic phase transitions and second-order statistics studied by Shiino's perturbation theory and strongly nonlinear Smoluchowski equations. AB - We study the first- and second-order statistical properties of a dynamical Maier Saupe model for liquid crystals that is given in terms of a nonlinear Smoluchowski equation. Using Shiino's perturbation theory, we analyze the first order statistics and give a rigorous proof of the emergence of a phase transition from a uniform distribution to a nonuniform distribution, reflecting phase transitions from isotropic to nematic phases, as observed in nematic liquid crystals. Using the concept of strongly nonlinear Fokker-Planck equations, the second-order statistics of the dynamical Maier-Saupe model is studied and an analytical expression for the short-time autocorrelation function of the orientation of the crystal molecules is derived. PMID- 16383399 TI - Self-assembly of biaxial ordering and molecular tilt angle of chiral smectic liquid crystals in homeotropically aligned cells investigated using infrared spectroscopy. AB - Temperature dependences of the infrared absorbance have been measured for the four chiral liquid crystal samples in the homeotropic cell configuration. It is shown that the values of the orientational order parameter obtained using this method exhibit a remarkable similarity to the x-ray diffraction results of the smectic layer spacing and lead to accurate values of the molecular tilt angle. This has important consequences for the existing interpretation of the x-ray data. The proposed method, in many cases, may be considered as a valuable alternative to the x-ray diffraction, giving additional, important information about the orientations and the ordering of the molecular fragments. It is found that if the experimentally obtained order parameter is low, then the molecular biaxiality is exceptionally large. The average phenyl ring plane is found to lie close to the molecular tilt plane. PMID- 16383400 TI - Discrete flexoelectric polarizations and biaxial subphases with periodicities other than three and four layers in chiral smectic liquid crystals frustrated between ferroelectricity and antiferroelectricity. AB - The subphase for the temperature range that lies in between Sm-C(A)* and the three-layer Sm-C(A)* (1/3)subphase has been confirmed to exist using the measurements of electric-field-induced birefringence, optical rotation, and the characteristic reflection bands in the antiferroelectric liquid-crystalline compound, 1-trifluoromethylundecyl-4-(4'-dodecyloxybiphenyl-4-yl-carbonyloxy)-3 fluorobenzoate (12BIMF10). The measurements of electric-field-induced birefringence and optical rotatory power are made on -thick homeotropic cells, and the characteristic reflection bands are observed in free-standing films of thicknesses ranging from 30 to 50 microm. Several binary mixtures have been prepared by mixing (S)-12BIMF10 with (S)-4-(1-methylheptyloxycarbonyl)-phenyl-4' octylbiphenyl-4-carboxylate (MHPBC), and the effect of racemization of the compound on the character of the biaxial subphase (other than three and four layers) is also discussed. The results are interpreted in terms of the Emelyanenko-Osipov model [Phys. Rev. E 68, 051703 (2003)]; the effective long range couplings between the director orientations in separated smectic layers emerge after the minimization of free energy with respect to the total (ordinary spontaneous and discrete flexoelectric) polarizations, lifting the degeneracy and producing the nonplanar structures of the subphases. PMID- 16383401 TI - Bifurcation analysis and computer simulation of biaxial liquid crystals. AB - We extend the analysis of a mean-field model for biaxial liquid crystals recently proposed by Sonnet et al. [Phys. Rev. E 67, 061701 (2003)]. In particular, we perform a bifurcation analysis of the equilibrium equations and derive the complete phase diagram. We show that two order parameters suffice to label all equilibrium phases, though they exhibit different bifurcation patterns. A Monte Carlo simulation study is performed as well, confirming qualitatively the predictions of this analysis. PMID- 16383402 TI - Light-scattering study on the shear-orientation coupling of liquids near isotropic-to-nematic phase transition. AB - Microscopic interaction between the translation and rotation of rodlike molecules yields the shear-orientation coupling in the macroscopic view. The depolarized scattering component arising from orientational fluctuations gives a constant in the strength of the coupling, which is related to the three viscosity parameters introduced by de Gennes. A high-resolution system of light scattering with an improved optical beating method was utilized for measurements in the 4-cyano-4' alkylbiphenyl nCB series: propyl (3CB), butyl (4CB), pentyl (5CB), hexyl (6CB), heptyl (7CB), and octyl (8CB). All the coupling constants obtained in these samples showed clear dependence on temperature, and their behaviors were interpreted in terms of a critical phenomenon near T*, the virtual isotropic nematic phase transition point. The values of T* were also given by the light scattering experiment, and the critical exponent of shear-orientation coupling was determined to be 0.31. Furthermore, the characteristic value of the coupling constant was correlated with the aspect ratio of these rodlike molecules, and two clear facts were found: a stronger coupling in longer molecules; and an odd-even effect with respect to the number of C-C bonds in the alkyl chain. PMID- 16383403 TI - Reexamination of the Helfrich-Hurault effect in smectic-A liquid crystals. AB - The Helfrich-Hurault effect is a phase transition that occurs in samples of cholesteric or smectic liquid crystals subject to external electric or magnetic fields. In this paper we analyze the Helfrich-Hurault effect of smectic-A liquid crystals in an electrostatic field taking into account the complete electromechanical coupling. A comparison is made with the results already obtained for the partially coupled case where one takes into account only the effect of the field on the crystal configuration and considering that field unaffected. PMID- 16383404 TI - Structure of mesogens possessing B7 textures: the case of the bent-core mesogen 8 OPIMB-NO2. AB - A structural investigation of the compound 8-OPIMB-NO2 is carried out. This material is a mesogen formed by bent-core molecules where the so-called B7 phase was originally proposed. X-ray data are successfully indexed in terms of a oblique two-dimensional lattice. Using additional information from optical second harmonic generation several possible structural models are proposed. The models are based on molecular packing considerations. All these structures are clearly different from those of other materials previously classified as B7. The necessity of a revised nomenclature for classifying the structures of these compounds is pointed out. PMID- 16383405 TI - Aggregation behavior and chromonic liquid crystal properties of an anionic monoazo dye. AB - X-ray scattering and various optical techniques are utilized to study the aggregation process and chromonic liquid crystal phase of the anionic monoazo dye Sunset Yellow FCF. The x-ray results demonstrate that aggregation involves pi-pi stacking of the molecules into columns, with the columns undergoing a phase transition to an orientationally ordered chromonic liquid crystal phase at high dye concentration. Optical absorption measurements on dilute solutions reveal that the aggregation takes place at all concentrations, with the average aggregation number increasing with concentration. A simple theory based on the law of mass action and an isodesmic aggregation process is in excellent agreement with the experimental data and yields a value for the "bond" energy between molecules in an aggregate. Measurements of the birefringence and order parameter are also performed as a function of temperature in the chromonic liquid crystal phase. The agreement between these results and a more complicated theory of aggregation is quite reasonable. Overall, these results both confirm that the aggregation process for some dyes is isodesmic and provide a second example of a well-characterized chromonic system. PMID- 16383406 TI - Optical characterization of the nematic lyotropic chromonic liquid crystals: light absorption, birefringence, and scalar order parameter. AB - We report on the optical properties of the nematic (N) phase formed by lyotropic chromonic liquid crystals (LCLCs) in well aligned planar samples. LCLCs belong to a broad class of materials formed by one-dimensional molecular self-assembly and are similar to other systems such as "living polymers" and "wormlike micelles." We study three water soluble LCLC forming materials: disodium chromoglycate, a derivative of indanthrone called Blue 27, and a derivative of perylene called Violet 20. The individual molecules have a planklike shape and assemble into rodlike aggregates that form the phase once the concentration exceeds about 0.1 M. The uniform surface alignment of the N phase is achieved by buffed polyimide layers. According to the light absorption anisotropy data, the molecular planes are on average perpendicular to the aggregate axes and thus to the nematic director. We determined the birefringence of these materials in the N and biphasic N-isotropic (I) regions and found it to be negative and significantly lower in the absolute value as compared to the birefringence of typical thermotropic low-molecular-weight nematic materials. In the absorbing materials Blue 27 and Violet 20, the wavelength dependence of birefringence is nonmonotonic because of the effect of anomalous dispersion near the absorption bands. We describe positive and negative tactoids formed as the nuclei of the new phase in the biphasic N-I region (which is wide in all three materials studied). Finally, we determined the scalar order parameter of the phase of Blue 27 and found it to be relatively high, in the range 0.72-0.79, which puts the finding into the domain of general validity of the Onsager model. However, the observed temperature dependence of the scalar order parameter points to the importance of factors not accounted for in the athermal Onsager model, such as interaggregate interactions and the temperature dependence of the aggregate length. PMID- 16383407 TI - Nonstandard electroconvection in a bent-core nematic liquid crystal. AB - We characterize three nonstandard electrohydrodynamic instabilities in nematic liquid crystals composed of bent-core molecules. In addition to their shape, another important attribute of this material is that the anisotropy in the electrical conductivity changes sign as the frequency of the applied electric field changes. These instabilities do not appear to fit within the standard model for electroconvection. The first instability creates a pattern with stripes parallel to the initial director orientation, with a wavelength about equal to the separation of the cell plates. The next is the previously reported prewavy instability. The third instability is optically and dynamically identical to the prewavy instability, but is distinguished by different threshold behavior. PMID- 16383408 TI - Experimental determination of the full Landau potential of bent-core doped ferroelectric liquid crystals. AB - The full Landau potential of a ferroelectric liquid crystal, doped with two different bent-core molecules at varying concentration, was determined experimentally. Using a multicurve fitting procedure, temperature and electric field dependent tilt angle and polarization measurements were analyzed according to the generalized Landau model of ferroelectric liquid crystals. From this analysis the three Landau coefficients as well as the polarization-tilt coupling parameters were obtained as a function of dopant concentration. It is shown that the two most varied parameters are those of the first Landau coefficient and the (chiral) linear polarization-tilt coupling constant. The results quantitatively verify the chiral induction capability of nonchiral bent-core shaped dopants and an observed increase in the electroclinic effect with increasing dopant concentration. PMID- 16383409 TI - Dynamic self-consistent field theory for unentangled homopolymer fluids. AB - We present a lattice formulation of a dynamic self-consistent field (DSCF) theory that is capable of resolving interfacial structure, dynamics, and rheology in inhomogeneous, compressible melts and blends of unentangled homopolymer chains. The joint probability distribution of all the Kuhn segments in the fluid, interacting with adjacent segments and walls, is approximated by a product of one body probabilities for free segments interacting solely with an external potential field that is determined self-consistently. The effect of flow on ideal chain conformations is modeled with finitely extensible, nonlinearly elastic dumbbells in the Peterlin approximation, and related to stepping probabilities in a random walk. Free segment and stepping probabilities generate statistical weights for chain conformations in a self-consistent field, and determine local volume fractions of chain segments. Flux balance across unit lattice cells yields mean field transport equations for the evolution of free segment probabilities and of momentum densities on the Kuhn length scale. Diffusive and viscous contributions to the fluxes arise from segmental hops modeled as a Markov process, with transition rates reflecting changes in segmental interaction, kinetic energy, and entropic contributions to the free energy under flow. We apply the DSCF equations to study both transient and steady-state interfacial structure, flow, and rheology in a sheared planar channel containing either a one component melt or a phase-separated, two-component blend. PMID- 16383410 TI - End-to-end distance vector distribution with fixed end orientations for the wormlike chain model. AB - We find exact expressions for the end-to-end distance vector distribution function with fixed end orientations for the wormlike chain model. This function in Fourier-Laplace space adopts the form of infinite continued fractions, which emerges upon exploiting the hierarchical structure of the moment-based expansion. Our results are used to calculate the root-mean-square end displacement in a given direction for a chain with both end orientations fixed. We find that the crossover from rigid to flexible chains is marked by the root-mean-square end displacement slowly losing its angular dependence as the coupling between chain conformation and end orientation wanes. However, the coupling remains strong even for relatively flexible chains, suggesting that the end orientation strongly influences chain conformation for chains that are several persistence lengths long. We then show the behavior of the distribution function by a density plot of the probability as a function of the end-to-end distance vector for a wormlike chain in two dimensions with one end pointed in a fixed direction and the other end free (in its orientation). As we progress from high to low rigidity, the distribution function shifts from being peaked at a location near the full contour length of the chain in the forward direction, corresponding to a straight configuration, to being peaked near zero end separation, as in the Gaussian limit. The function exhibits double peaks in the crossover between these limiting behaviors. PMID- 16383411 TI - Nonlinear elasticity of single collapsed polyelectrolytes. AB - Nonlinear elastic responses of short and stiff polyelectrolytes are investigated by dynamic simulations on a single-molecule level. When a polyelectrolyte condensate undergoes a mechanical unfolding, two types of force-extension curves- i.e., a force plateau and a stick-release pattern--are observed depending on the strength of the electrostatic interaction. We provide a physical interpretation of such force-extension behavior in terms of intramolecular structures of the condensates. We also describe charge distributions of counterions condensed onto a polyelectrolyte, which clarify formation of one-dimensional strongly correlated liquid at large Coulomb coupling regime. These findings may provide significant insights into the relationship between a molecular elasticity and a molecular mechanism of like-charge attractions observed in a wide range of charged biopolymer systems. PMID- 16383412 TI - Scattering functions of knotted ring polymers. AB - We discuss the scattering function of a Gaussian random polygon with N nodes under a given topological constraint through simulation. We evaluate the form factor PK(q) of a Gaussian polygon of N = 200 having a fixed knot K for some different knots such as the trivial, trefoil, and figure-eight knots. Here the Gaussian polygons with different knots K have distinct values of the mean-square radius of gyration, R2(G,K). We obtain the Kratky plots of the form factors- i.e., the plots of (qR(G,K))2PK(q) versus qR(G,K)--for the different topological constraints and discuss nontrivial large-q behavior as well as small-q behavior for the scattering functions. We also find that the distinct values of R2(G,K) play an important role in the large-q and small-q properties of the Kratky plots. PMID- 16383413 TI - Hydrodynamic theories for mixtures of polymers and rodlike liquid crystalline polymers. AB - We develop a hydrodynamic theory for flows of incompressible blends of flexible polymers and rodlike nematic polymers (RNPs) or rodlike nematic liquid crystal polymers (RNLCPs) extending the thermodynamical theory of Muratov and E [J. Chem. Phys. 116, 4723 (2002)] for phase separation kinetics of the blend. We model the flexible polymer molecules in the polymer matrix as Rouse chains and assume the translational diffusion of the molecules is predominantly through the volume fraction of the flexible polymer and the molecules of rodlike nematic liquid crystal polymers. We then (i) derive the translational flux for the rodlike nematic liquid crystal polymers to ensure the incompressibility constraint; (ii) derive the elastic stress tensor, accounting for the contribution from both the rodlike nematic polymer and the flexible polymer matrix, as well as the extra elastic body force due to the nonlocal intermolecular potential for long range molecular interaction; (iii) show that the theory obeys positive entropy production and thereby satisfies the second law of thermodynamics. By applying the gradient expansion technique on the number density function of RNLCPs, we present an approximate, weakly nonlocal theory in differential form in which the intermolecular potential is given by gradients of the number density function of the RNLCP and the volume fraction of the flexible polymer. In the approximate theory, the elastic stress is augmented by an extra stress tensor due to the spatial convection of the macroscopic material point and long range interaction, whose divergence yields the analogous extra elastic body force with respect to the nonlocal intermolecular potential. Finally, we compare the model in steady simple shear with the Doi theory for bulk monodomains of rodlike nematic polymers. PMID- 16383415 TI - Arbitrary nonlinearities in convective population dynamics with small diffusion. AB - Convective counterparts of variants of the nonlinear Fisher equation which describes reaction diffusion systems in population dynamics are studied with the help of an analytic prescription and shown to lead to interesting consequences for the evolution of population densities. The initial-value problem is solved explicitly for some cases, and for others it is shown how to find traveling-wave solutions analytically. The effect of adding diffusion to the convective equations is first studied through exact analysis through a piecewise linear representation of the nonlinearity. Using an appropriate small parameter suggested by that analysis, a perturbative treatment is developed to treat the case in which the convective evolution is augmented by a small amount of diffusion. PMID- 16383414 TI - Fluctuations and instability of a biological membrane induced by interaction with macromolecules. AB - This paper studies the dynamics and fluctuations of a phospholipidic membrane in the presence of a diffusion field of foreign molecules, such as polymers, proteins, etc., which have the ability to adsorb on, and to desorb from, the membrane. We develop a model that includes, besides hydrodynamics, molecular diffusion in the surrounding fluid and lateral diffusion (i.e., diffusion along the membrane) as well as the kinetics of attachment and detachment to and from the membrane. This model is exploited here for the case of a free membrane which is globally at equilibrium while the nonequilibrium part will be presented in the future. We show that if the coupling between the membrane and the molecules is strong enough, the flat membrane can suffer a morphological instability. The numerical calculation in the nonlinear regime reveals budding, and an initial stage of spontaneous vesicle emission. When the condition of stability is satisfied we show how kinetic fluctuations lead to a rich variety of dynamical behaviors expressing the dominant dissipation mechanisms. We show that in the limit of well separated length scales related to the physical mechanisms that enter into play, the width of the membrane fluctuations exhibits various dynamical scalings with universal scaling exponents. It is shown that the usual behavior with time w approximately t(1/3) is altered in various time and length scales of interest. For example, we find that , w approximately t(1/4), w approximately t(1/2), and w approximately [t ln(t)](1/2), depending on which physical mechanism limits the membrane fluctuations on the time scale under consideration. The experimental study of the fluctuation spectrum can be viewed as a precious tool in order to have access to the underlying microscopic physical mechanisms. PMID- 16383416 TI - Role of network dynamics in shaping spike timing reliability. AB - We study the reliability of cortical neuron responses to periodically modulated synaptic stimuli. Simple map-based models of two different types of cortical neurons are constructed to replicate the intrinsic resonances of reliability found in experimental data and to explore the effects of those resonance properties on collective behavior in a cortical network model containing excitatory and inhibitory cells. We show that network interactions can enhance the frequency range of reliable responses and that the latter can be controlled by the strength of synaptic connections. The underlying dynamical mechanisms of reliability enhancement are discussed. PMID- 16383417 TI - Model for complex heart rate dynamics in health and diseases. AB - A physiologically motivated, dynamical model of cardiovascular autonomic regulation is shown to be capable of generating long-range correlated and multifractal heart rate. Virtual disease simulations are carried out systematically to account for the disease-induced relative dysfunction of the parasympathetic and the sympathetic branches of the autonomic control. Statistical agreement of the simulation results with those of real life data is reached, suggesting the possible use of the model as a state-of-the-art basis for further understanding of the physiological correlates of complex heart rate dynamics. PMID- 16383418 TI - Apparent persistence length renormalization of bent DNA. AB - We derive the single molecule equation of state (force-extension relation) for DNA molecules bearing sliding loops and deflection defects. Analytical results are obtained in the large force limit by employing an analogy with instantons in quantum mechanical tunneling problems. The results reveal a remarkable feature of sliding loops--an apparent strong reduction of the persistence length. We generalize these results to several other experimentally interesting situations ranging from rigid DNA-protein loops to the problem of anchoring deflections in atomic force microscopy stretching of semiflexible polymers. Expressions relating the force-extension measurements to the underlying loop or boundary deflection geometry are provided and applied to the case of the gal repressor dimer protein. The theoretical predictions are complemented and quantitatively confirmed by molecular dynamics simulations. PMID- 16383419 TI - Instability of defensive alliances in the predator-prey model on complex networks. AB - A model of six-species food web is studied in the viewpoint of spatial interaction structures. Each species has two predators and two preys, and it was previously known that the defensive alliances of three cyclically predating species self-organize in two dimensions. The alliance-breaking transition occurs as either the mutation rate is increased or interaction topology is randomized in the scheme of the Watts-Strogatz model. In the former case of temporal disorder, via the finite-size scaling analysis, the transition is clearly shown to belong to the two-dimensional Ising universality class. In contrast, the geometric or spatial randomness for the latter case yields a discontinuous phase transition. The mean-field limit of the model is analytically solved and then compared with numerical results. The dynamic universality and the temporally periodic behaviors are also discussed. PMID- 16383420 TI - Tumor growth instability and the onset of invasion. AB - Motivated by experimental observations, we develop a mathematical model of chemotactically directed tumor growth. We present an analytical study of the model as well as a numerical one. The mathematical analysis shows that: (i) tumor cell proliferation by itself cannot generate the invasive branching behavior observed experimentally, (ii) heterotype chemotaxis provides an instability mechanism that leads to the onset of tumor invasion, and (iii) homotype chemotaxis does not provide such an instability mechanism but enhances the mean speed of the tumor surface. The numerical results not only support the assumptions needed to perform the mathematical analysis but they also provide evidence of (i), (ii), and (iii). Finally, both the analytical study and the numerical work agree with the experimental phenomena. PMID- 16383421 TI - Three-bead rotating chain model shows universality in the stretching of proteins. AB - We introduce a model of proteins in which all of the key atoms in the protein backbone are accounted for, thus extending the freely rotating chain model. We use average bond lengths and average angles from the Protein Data Bank as input parameters, leaving the number of residues as a single variable. The model is used to study the stretching of proteins in the entropic regime. The results of our Monte Carlo simulations are found to agree well with experimental data, suggesting that the force extension plot is universal and does not depend on the side chains or primary structure of the proteins. PMID- 16383422 TI - Solving the advection-diffusion equations in biological contexts using the cellular Potts model. AB - The cellular Potts model (CPM) is a robust, cell-level methodology for simulation of biological tissues and morphogenesis. Both tissue physiology and morphogenesis depend on diffusion of chemical morphogens in the extra-cellular fluid or matrix (ECM). Standard diffusion solvers applied to the cellular potts model use finite difference methods on the underlying CPM lattice. However, these methods produce a diffusing field tied to the underlying lattice, which is inaccurate in many biological situations in which cell or ECM movement causes advection rapid compared to diffusion. Finite difference schemes suffer numerical instabilities solving the resulting advection-diffusion equations. To circumvent these problems we simulate advection diffusion within the framework of the CPM using off-lattice finite-difference methods. We define a set of generalized fluid particles which detach advection and diffusion from the lattice. Diffusion occurs between neighboring fluid particles by local averaging rules which approximate the Laplacian. Directed spin flips in the CPM handle the advective movement of the fluid particles. A constraint on relative velocities in the fluid explicitly accounts for fluid viscosity. We use the CPM to solve various diffusion examples including multiple instantaneous sources, continuous sources, moving sources, and different boundary geometries and conditions to validate our approximation against analytical and established numerical solutions. We also verify the CPM results for Poiseuille flow and Taylor-Aris dispersion. PMID- 16383423 TI - Velocity half-sphere model for multiple light scattering in turbid media. AB - We extend the traditional diffusion theory by distinguishing between the energy radiance in the forward and backward directions at each point in space. This approach leads to a new effective source for the diffusion equation that is nonzero for an anisotropic light source. It differs significantly from the diffusion theory for short source-detector spacings. We derive an analytical solution for the two lowest-order velocity moments of the radiance. PMID- 16383424 TI - Automatic network coupling analysis for dynamical systems based on detailed kinetic models. AB - We introduce a numerical complexity reduction method for the automatic identification and analysis of dynamic network decompositions in (bio)chemical kinetics based on error-controlled computation of a minimal model dimension represented by the number of (locally) active dynamical modes. Our algorithm exploits a generalized sensitivity analysis along state trajectories and subsequent singular value decomposition of sensitivity matrices for the identification of these dominant dynamical modes. It allows for a dynamic coupling analysis of (bio)chemical species in kinetic models that can be exploited for the piecewise computation of a minimal model on small time intervals and offers valuable functional insight into highly nonlinear reaction mechanisms and network dynamics. We present results for the identification of network decompositions in a simple oscillatory chemical reaction, time scale separation based model reduction in a Michaelis-Menten enzyme system and network decomposition of a detailed model for the oscillatory peroxidase-oxidase enzyme system. PMID- 16383425 TI - Model study of prionlike folding behavior in aggregated proteins. AB - We investigate the folding behavior of protein sequences by numerically studying all sequences with a maximally compact lattice model through exhaustive enumeration. We get the prionlike behavior of protein folding. Individual proteins remaining stable in the isolated native state may change their conformations when they aggregate. We observe the folding properties as the interfacial interaction strength changes and find that the strength must be strong enough before the propagation of the most stable structures happens. PMID- 16383426 TI - Retrieval-time properties of the Little-Hopfield model and their physiological relevance. AB - We perform an extensive numerical investigation on the retrieval dynamics of the synchronous Hopfield model, also known as Little-Hopfield model, up to sizes of 2(18) neurons. Our results correct and extend much of the early simulations on the model. We find that the average convergence time has a power law behavior for a wide range of system sizes, whose exponent depends both on the network loading and the initial overlap with the memory to be retrieved. Surprisingly, we also find that the variance of the convergence time grows as fast as its average, making it a non-self-averaging quantity. Based on the simulation data we differentiate between two definitions for memory retrieval time, one that is mathematically strict, tau(c), the number of updates needed to reach the attractor whose properties we just described, and a second definition correspondent to the time tau(eta) when the network stabilizes within a tolerance threshold eta such that the difference of two consecutive overlaps with a stored memory is smaller that eta. We show that the scaling relationships between tau(c) and tau(eta) and the typical network parameters as the memory load alpha or the size of the network N vary greatly, being tau(eta) relatively insensitive to system sizes and loading. We propose tau(eta) as the physiological realistic measure for the typical attractor network response. PMID- 16383427 TI - Microrheological detection of protein unfolding. AB - We apply passive probes to protein solutions and evaluate the viscous response to folding and unfolding, allowing us to accurately quantify both the thermodynamics of protein folding and the structural dimensions of the protein molecules with subnanometer resolution. Hard-sphere approximations predict a measurable change in relative viscosity as the hydrodynamic volume fraction of protein molecules increases during unfolding. Microrheology measures these changes to unambiguously evaluate the ensemble average characteristics of the unfolded state in a denaturant, urea, while minimizing the shear-induced unfolding and alignment associated with conventional rheometry. PMID- 16383428 TI - Hierarchical structure analysis describing abnormal base composition of genomes. AB - Abnormal base compositional patterns of genomic DNA sequences are studied in the framework of a hierarchical structure (HS) model originally proposed for the study of fully developed turbulence [She and Leveque, Phys. Rev. Lett. 72, 336 (1994)]. The HS similarity law is verified over scales between 10(3)bp and 10(5)bp, and the HS parameter beta is proposed to describe the degree of heterogeneity in the base composition patterns. More than one hundred bacteria, archaea, virus, yeast, and human genome sequences have been analyzed and the results show that the HS analysis efficiently captures abnormal base composition patterns, and the parameter beta is a characteristic measure of the genome. Detailed examination of the values of beta reveals an intriguing link to the evolutionary events of genetic material transfer. Finally, a sequence complexity (S) measure is proposed to characterize gradual increase of organizational complexity of the genome during the evolution. The present study raises several interesting issues in the evolutionary history of genomes. PMID- 16383429 TI - Morphogen gradient formation in a complex environment: an anomalous diffusion model. AB - Current models of morphogen-induced patterning assume that morphogens undergo normal, or Fickian, diffusion, although the validity of this assumption has never been examined. Here we argue that the interaction of morphogens with the complex extracellular surrounding may lead to anomalous diffusion. We present a phenomenological model that captures this interaction, and derive the properties of the morphogen profile under conditions of anomalous (non-Fickian) diffusion. In this context we consider the continuous time random walk formalism and extend its application to account for degradation of morphogen particles. We show that within the anomalous diffusion model, morphogen profiles are fundamentally distinct from the corresponding Fickian profiles. Differences were found in several key aspects, including the role of degradation in determining the profile, the rate by which it spreads in time and its long-term behavior. We analyze the effect of an abrupt change in the extracellular environment on the concentration profiles. Furthermore, we discuss the robustness of the morphogen distribution to fluctuations in morphogen production rate, and describe a feedback mechanism that can buffer such fluctuations. Our study also provides rigorous criteria to distinguish experimentally between Fickian and anomalous modes of morphogen transport. PMID- 16383430 TI - Segmentation algorithm for DNA sequences. AB - A new measure, to quantify the difference between two probability distributions, called the quadratic divergence, has been proposed. Based on the quadratic divergence, a new segmentation algorithm to partition a given genome or DNA sequence into compositionally distinct domains is put forward. The new algorithm has been applied to segment the 24 human chromosome sequences, and the boundaries of isochores for each chromosome were obtained. Compared with the results obtained by using the entropic segmentation algorithm based on the Jensen-Shannon divergence, both algorithms resulted in all identical coordinates of segmentation points. An explanation of the equivalence of the two segmentation algorithms is presented. The new algorithm has a number of advantages. Particularly, it is much simpler and faster than the entropy-based method. Therefore, the new algorithm is more suitable for analyzing long genome sequences, such as human and other newly sequenced eukaryotic genome sequences. PMID- 16383431 TI - Elasticity of semiflexible polymers in two dimensions. AB - We study theoretically the entropic elasticity of a semiflexible polymer, such as DNA, confined to two dimensions. Using the worm-like-chain model we obtain an exact analytical expression for the partition function of the polymer pulled at one end with a constant force. The force-extension relation for the polymer is computed in the long chain limit in terms of Mathieu characteristic functions. We also present applications to the interaction between a semiflexible polymer and a nematic field, and derive the nematic order parameter and average extension of the polymer in a strong field. PMID- 16383432 TI - Statistical physics models for nacre fracture simulation. AB - Natural biological materials such as nacre (or mother-of-pearl), exhibit phenomenal fracture strength and toughness properties despite the brittle nature of their constituents. For example, nacre's work of fracture is three orders of magnitude greater than that of a single crystal of its constituent mineral. This study investigates the fracture properties of nacre using a simple discrete lattice model based on continuous damage random thresholds fuse network. The discrete lattice topology of the proposed model is based on nacre's unique brick and mortar microarchitecture, and the mechanical behavior of each of the bonds in the discrete lattice model is governed by the characteristic modular damage evolution of the organic matrix that includes the mineral bridges between the aragonite platelets. The analysis indicates that the excellent fracture properties of nacre are a result of their unique microarchitecture, repeated unfolding of protein molecules (modular damage evolution) in the organic polymer, and the presence of fiber bundle of mineral bridges between the aragonite platelets. The numerical results obtained using this simple discrete lattice model are in excellent agreement with the previously obtained experimental results, such as nacre's stiffness, tensile strength, and work of fracture. PMID- 16383433 TI - Pattern formation on the surface of cationic-anionic cylindrical aggregates. AB - Charged pattern formation on the surfaces of self-assembled cylindrical micelles formed from oppositely charged heterogeneous molecules such as cationic and anionic peptide amphiphiles is investigated. The net incompatibility chi among different components results in the formation of segregated domains, whose growth is inhibited by electrostatics. The transition to striped phases proceeds through an intermediate structure governed by fluctuations, followed by states with various lamellar orientations, which depend on cylinder radius Rc and chi. We analyze the specific heat, susceptibility S(q*), domain size Lambda = 2pi/q*, and morphology as a function of Rc and chi. PMID- 16383434 TI - Phase-field approach to three-dimensional vesicle dynamics. AB - We extend our recent phase-field [T. Biben and C. Misbah, Phys. Rev. E 67, 031908 (2003)] approach to 3D vesicle dynamics. Unlike the boundary-integral formulations, based on the use of the Oseen tensor in the small Reynolds number limit, this method offers several important flexibilities. First, there is no need to track the membrane position; rather this is automatically encoded in dynamics of the phase field to which we assign a finite width representing the membrane extent. Secondly, this method allows naturally for any topology change, like vesicle budding, for example. Thirdly, any non-Newtonian constitutive law, that is generically nonlinear, can be naturally accounted for, a fact which is precluded by the boundary integral formulation. The phase-field approach raises, however, a complication due to the local membrane incompressibility, which, unlike usual interfacial problems, imposes a nontrivial constraint on the membrane. This problem is solved by introducing dynamics of a tension field. The first purpose of this paper is to show how to write adequately the advected-field model for 3D vesicles. We shall then perform a singular expansion of the phase field equation to show that they reduce, in the limit of a vanishing membrane extent, to the sharp boundary equations. Then, we present some results obtained by the phase-field model. We consider two examples; (i) kinetics towards equilibrium shapes and (ii) tanktreading and tumbling. We find a very good agreement between the two methods. We also discuss briefly how effects, such as the membrane shear elasticity and stretching elasticity, and the relative sliding of monolayers, can be accounted for in the phase-field approach. PMID- 16383435 TI - Tradeoff between short-term and long-term adaptation in a changing environment. AB - We investigate the competition dynamics of two microbial or viral strains that live in an environment that switches periodically between two states. One of the strains is adapted to the long-term environment, but pays a short-term cost, while the other is adapted to the short-term environment and pays a cost in the long term. We explore the tradeoff between these alternative strategies in extensive numerical simulations and present a simple analytic model that can predict the outcome of these competitions as a function of the mutation rate and the time scale of the environmental changes. Our model is relevant for arboviruses, which alternate between different host species on a regular basis. PMID- 16383436 TI - Density functional theory approach for coarse-grained lipid bilayers. AB - Lipid bilayers are important inhomogeneous fluid systems that mediate the environment of cells and the interaction of cells with their environment. A variety of approaches have been taken to model the lipid molecules in bilayers, from all atom molecular dynamics to rigid body liquid crystals. In this paper we discuss the application of a density functional theory approach that treats the lipid molecules at the coarse-grained level of a freely jointed chain. This approach allows for compressibility effects, and can therefore be used to study not only the long range structure in lipid bilayers, but also the nanoscale structure induced in the bilayer when the lipids crystallize or when an inclusion (e.g., an embedded protein) is present. This paper presents a detailed analysis of fluid bilayers and lamellae predicted by the theory. In particular we locate solutions with zero surface tension. We calculate the phase diagram for all possible phases with planar symmetry, including uniform macrophases. Surprisingly, we find a first-order phase transition from the lamellar phase to an isolated bilayer phase on lowering the temperature. This transition appears to be driven by solvent packing effects. A further lowering of the temperature leads to a set of highly ordered bilayers. PMID- 16383437 TI - Comparison of density functional theory and simulation of fluid bilayers. AB - We compare results of classical density functional theory (DFT) to molecular dynamics (MD) simulations of coarse-grained models of lipids in solvent. We find that the DFT captures the liquid structure of coarse-grained lipids both near surfaces and in bilayers adequately. In contrast we find that the MD simulations do not predict ordering in bilayers as is observed in low temperature DFT calculations. The mechanical properties of the fluid DFT bilayers are qualitatively similar to those of the MD bilayers; in particular the shapes of the lateral stress profiles are similar. Values of the area compressibility modulus are in reasonable agreement with previous work on coarse-grained lipids. PMID- 16383438 TI - Simplified dynamic model for the motility of irregular echinoids. AB - Inspired by the locomotion mechanism of sea urchins, we study the locomotion of an irregular echinoid by means of a simplified dynamical model. We prove that if two conjectures are assumed, the geometrical arrangement of the five ambulacral petals of irregular echinoids should form a eutactic star in order to optimize motility. We firstly propose an adequate "measure" of eutacticity that allows us to to verify the statistical tendency to such a property for a representative collection of fossil specimens. Next, regarding dynamics, the biological advantage of eutactic stars is addressed as a minimal path problem. Finally, we study the stability of some eutactic stars under small perturbations. PMID- 16383439 TI - Electromechanical effects on tether formation from lipid membranes: a theoretical analysis. AB - The material properties of biomembranes can be measured by forming a tether, a thin bilayer tube that extends from the membrane surface. Recent experiments have demonstrated that the force required to maintain a tether is sensitive to the transmembrane potential. As a first approach towards understanding this phenomenon, a thermodynamic analysis of the influence of electrical fields on tether formation from an aspirated lipid vesicle is developed. The analysis considers contributions from Maxwell stresses as well as two forms of electromechanical coupling: coupling between the electric field and curvature strain (flexoelectric coupling) and between the electric field and areal strain (piezoelectric coupling). Predictions of equilibrium tether conformations are obtained numerically. For expected values of the dimensionless coupling parameters, flexoelectric coupling alters the force required to form a tether of a given length, while piezoelectric coupling and Maxwell forces do not greatly change the force versus tether length behavior. The results of this analysis indicate that tether experiments have the potential to characterize electromechanical coupling in both synthetic and cellular membranes. PMID- 16383440 TI - Nonequilibrium statistical mechanical models for cytoskeletal assembly: towards understanding tensegrity in cells. AB - The cytoskeleton is not an equilibrium structure. To develop theoretical tools to investigate such nonequilibrium assemblies, we study a statistical physical model of motorized spherical particles. Though simple, it captures some of the key nonequilibrium features of the cytoskeletal networks. Variational solutions of the many-body master equation for a set of motorized particles accounts for their thermally induced Brownian motion as well as for the motorized kicking of the structural elements. These approximations yield stability limits for crystalline phases and for frozen amorphous structures. The methods allow one to compute the effects of nonequilibrium behavior and adhesion (effective cross-linking) on the mechanical stability of localized phases as a function of density, adhesion strength, and temperature. We find that nonequilibrium noise does not necessarily destabilize mechanically organized structures. The nonequilibrium forces strongly modulate the phase behavior and have comparable effect as the adhesion due to cross-linking. Modeling transitions such as these allows the mechanical properties of cytoskeleton to rapidly and adaptively change. The present model provides a statistical mechanical underpinning for a tensegrity picture of the cytoskeleton. PMID- 16383441 TI - Scale-free movement patterns arising from olfactory-driven foraging. AB - The scale-free movement patterns of some foragers has aroused considerable excitement because they are known to optimize the success of random searches when target sites are randomly and sparsely distributed. Here it is shown that scale free movement patterns can arise from a naive response of foragers to scent concentrations within turbulent plume structures. PMID- 16383442 TI - Viscoelastic Fitzhugh-Nagumo models. AB - An extended Fitzhugh-Nagumo model including linear viscoelasticity is derived in general and studied in detail in the one-dimensional case. The equations of the theory are numerically integrated in two situations: (i) a free insulated fiber activated by an initial Gaussian distribution of action potential, and (ii) a clamped fiber stimulated by two counter phased currents, located at both ends of the space domain. The former case accounts for a description of the physiological experiments on biological samples in which a fiber contracts because of the spread of action potential, and then relaxes. The latter case, instead, is introduced to extend recent models discussing a strongly electrically stimulated fiber so that nodal structures associated on quasistanding waves are produced. Results are qualitatively in agreement with physiological behavior of cardiac fibers. Modifications induced on the action potential of a standard Fitzhugh Nagumo model appear to be very small even when strong external electric stimulations are activated. On the other hand, elastic backreaction is evident in the model. PMID- 16383443 TI - Tube formation and spontaneous budding in a fluid charged membrane. AB - We determine the equations governing the equilibrium shape of an axially symmetric charged fluid membrane under the action of an external field. We consider that the charges are free to diffuse along the membrane, and we neglect the effect of screening counterions. By numerically integrating these equations for a membrane spread across a hole by a constant tension, we show that there exists a threshold electric field above which an infinitely long tube can be pulled. The threshold field for pulling a tube decreases as the surface charge density of the membrane increases, reaching zero at a finite critical value. Above this critical density of charge, the membrane spontaneously buds, in the absence of externally applied fields, because of the electrostatic repulsion of its charges. PMID- 16383444 TI - Structure and interaction in protein solutions as studied by small-angle neutron scattering. AB - Small-angle neutron scattering (SANS) measurements have been performed to compare the effect of the salts KF, KCl, and KBr on crystallization in aqueous solution of lysozyme protein. It is found that the propensity of the salt to crystallize protein follows the Hoffmeister series (KF or = 1/N the scaling parameter deviates from it significantly. For WS small-world networks, in the special region p(r) element of [0.05,0.2], typical scale invariance is found. For growing random networks, in the range of theta element of [0.33,049], we have delta = 0.6 +.- 0.1. And the value of delta oscillates around delta = 0.6 abruptly. In the range of delta element of [0.54,1], we have basically element of > 0.7. Scale invariance is one of the common features of the three kinds of networks, which can be employed as a global measurement of complex networks in a unified way. PMID- 16383481 TI - Wavelet analysis and scaling properties of time series. AB - We propose a wavelet based method for the characterization of the scaling behavior of nonstationary time series. It makes use of the built-in ability of the wavelets for capturing the trends in a data set, in variable window sizes. Discrete wavelets from the Daubechies family are used to illustrate the efficacy of this procedure. After studying binomial multifractal time series with the present and earlier approaches of detrending for comparison, we analyze the time series of averaged spin density in the 2D Ising model at the critical temperature, along with several experimental data sets possessing multifractal behavior. PMID- 16383482 TI - Escape or switching at short times. AB - In the standard Arrhenius picture [S. Arrhenius, Z. Phys. Chem., Stoechiom. Verwandtschaftsl. 4, 226 (1889); L. Neel, Ann. Geophys. (C.N.R.S.) 5, 99 (1949)] of thermal switching or escape from a metastable to a stable state, the escape probability per unit time P(s)(t) decreases monotonically with time t as P(s)(t) approximately e(-t/tau(D)), where the decay time tau(D) = tau0e(U/k(B)T), with U the energy barrier, k(B)T the thermal energy, and tau0 the time between escape attempts. Here, we extend the Arrhenius picture to shorter times by deriving general conditions under which P(s)(t) is peaked rather than monotonic, and showing that in the simplest scenario the peak time tau(P) diverges with tau(D) as ln(tau(D)). PMID- 16383483 TI - Structure and spectrum of anisotropically confined two-dimensional clusters with logarithmic interaction. AB - We studied the structural and spectral properties of a classical system consisting of a finite number of particles, moving in two dimensions, and interacting through a repulsive logarithmic potential and held together by an anisotropic harmonic potential. Increasing the anisotropy of the confinement potential can drive the system from a two-dimensional (2D) to a one-dimensional (1D) configuration. This change occurs through a sequence of structural transitions of first and second order which are reflected in the normal mode frequencies. Our results of the ground state configurations are compared with recent experiments and we obtained a satisfactory agreement. The transition from the 1D line structure to the 2D structure occurs through a zigzag transition which is of second order. We found analytical expressions for the eigenfrequencies before the zigzag transition, which allowed us to obtain an analytical expression for the anisotropy parameter at which the zigzag transition occurs as a function of the number of particles in the system. PMID- 16383484 TI - 1/d expansion for k-core percolation. AB - The physics of k-core percolation pertains to those systems whose constituents require a minimum number of k connections to each other in order to participate in any clustering phenomenon. Examples of such a phenomenon range from orientational ordering in solid ortho-para H2 mixtures to the onset of rigidity in bar-joint networks to dynamical arrest in glass-forming liquids. Unlike ordinary (k = 1) and biconnected (k = 2) percolation, the mean field k > or = 3 core percolation transition is both continuous and discontinuous, i.e., there is a jump in the order parameter accompanied with a diverging length scale. To determine whether or not this hybrid transition survives in finite dimensions, we present a 1/d expansion for k-core percolation on the d-dimensional hypercubic lattice. We show that to order 1/d3 the singularity in the order parameter and in the susceptibility occur at the same value of the occupation probability. This result suggests that the unusual hybrid nature of the mean field k-core transition survives in high dimensions. PMID- 16383485 TI - Scaling in critical random Boolean networks. AB - We derive mostly analytically the scaling behavior of the number of nonfrozen and relevant nodes in critical Kauffman networks (with two inputs per node) in the thermodynamic limit. By defining and analyzing a stochastic process that determines the frozen core we can prove that the mean number of nonfrozen nodes scales with the network size N as N(2/3), with only N(1/3) nonfrozen nodes having two nonfrozen inputs. We also show the probability distributions for the numbers of these nodes. Using a different stochastic process, we determine the scaling behavior of the number of relevant nodes. Their mean number increases for large N as N(1/3), and only a finite number of relevant nodes have two relevant inputs. It follows that all relevant components apart from a finite number are simple loops and that the mean number and length of attractors increases faster than any power law with network size. PMID- 16383486 TI - Fluctuation growth and saturation in nonlinear oscillators on the threshold of bifurcation of spontaneous symmetry breaking. AB - We study prebifurcation fluctuation amplification in nonlinear oscillators subject to bifurcations of spontaneous symmetry breaking which are manifest in the doubling of stable equilibrium states. Our theoretical estimates of both the linear growth and the nonlinear saturation of the fluctuations are in good agreement with our results from numerical simulations. We show that in the saturation mode, the fluctuation variance is proportional to the standard deviation of the external noise, whereas in the linear mode, the fluctuation variance is proportional to the noise variance. It is demonstrated that the phenomenon of prebifurcation noise amplification is more pronounced in the case of a slow transition through the bifurcation point. The amplification of fluctuations in this case makes it easier to form a symmetric probability of the final equilibrium states. In contrast, for a fast transition through the bifurcation point, the effect of amplification is much less pronounced. Under backward and forward passages through the bifurcation point, a loop of noise dependent hysteresis emerges here. We find that for a fast transition of the nonlinear oscillator through the bifurcation point, the probability symmetry of the final equilibrium states is destroyed. PMID- 16383487 TI - Simple beam model for the shear failure of interfaces. AB - We propose a model for the shear failure of a glued interface between two solid blocks. We model the interface as an array of elastic beams which experience stretching and bending under shear load and break if the two deformation modes exceed randomly distributed breaking thresholds. The two breaking modes can be independent or combined in the form of a von Mises-type breaking criterion. Assuming global load sharing following the beam breaking, we obtain analytically the macroscopic constitutive behavior of the system and describe the microscopic process of the progressive failure of the interface. We work out an efficient simulation technique which allows for the study of large systems. The limiting case of very localized interaction of surface elements is explored by computer simulations. PMID- 16383488 TI - Statistical analysis of 22 public transport networks in Poland. AB - Public transport systems in 22 Polish cities have been analyzed. The sizes of these networks range from N = 152 to 2881. Depending on the assumed definition of network topology, the degree distribution can follow a power law or can be described by an exponential function. Distributions of path lengths in all considered networks are given by asymmetric, unimodal functions. Clustering, assortativity, and betweenness are studied. All considered networks exhibit small world behavior and are hierarchically organized. A transition between dissortative small networks N approximately < or = 500 and assortative large networks N approximately > or = 500 is observed. PMID- 16383489 TI - Diffusion in scale-free networks with annealed disorder. AB - The scale-free (SF) networks that have been studied so far contained quenched disorder generated by random dilution which does not vary with the time. In practice, if a SF network is to represent, for example, the worldwide web, then the links between its various nodes may temporarily be lost and reestablished again later on. This gives rise to SF networks with annealed disorder. Even if the disorder is quenched, it may be more realistic to generate it by a dynamical process that is happening in the network. In this paper, we study diffusion in SF networks with annealed disorder generated by various scenarios, as well as in SF networks with quenched disorder which, however, is generated by the diffusion process itself. Several quantities of the diffusion process are computed, including the mean number of distinct sites visited, the mean number of returns to the origin, and the mean number of connected nodes that are accessible to the random walkers at any given time. The results, including (1) greatly reduced growth with the time of the mean number of distinct sites visited, (2) blocking of the random walkers, (3) the existence of a phase diagram that separates the region in which diffusion is possible from one in which diffusion is impossible, and (4) a transition in the structure of the networks at which the mean number of distinct sites visited vanishes, indicate completely different behavior for the computed quantities than those in SF networks with quenched disorder generated by simple random dilution. PMID- 16383490 TI - Partially asymmetric zero-range process with quenched disorder. AB - We consider the one-dimensional partially asymmetric zero-range process where the hopping rates as well as the easy direction of hopping are random variables. For this type of disorder there is a condensation phenomenon in the thermodynamic limit: the particles typically occupy one single site and the fraction of particles outside the condensate is vanishing. We use extreme value statistics and an asymptotically exact strong disorder renormalization group method to explore the properties of the steady state. In a finite system of L sites the current vanishes as J approximately L(-z), where the dynamical exponent, z, is exactly calculated. For 0 < z < 1 the transport is realized by N(a) approximately L(1-z) active particles, which move with a constant velocity, whereas for z > 1 the transport is due to the anomalous diffusion of a single Brownian particle. Inactive particles are localized at a second special site and their number in rare realizations is macroscopic. The average density profile of inactive particles has a width of xi approximately delta(-2) in terms of the asymmetry parameter delta. In addition to this, we have investigated the approach to the steady state of the system through a coarsening process and found that the size of the condensate grows as n(L) approximately t(1/(1+z)) for large times. For the unbiased model z is formally infinite and the coarsening is logarithmically slow. PMID- 16383491 TI - Analytical investigation of oscillations in intersecting flows of pedestrian and vehicle traffic. AB - In two intersecting many-particle streams, one can often find the emergence of oscillatory patterns. Here, we investigate the interaction of pedestrians with vehicles, when they try to cross a road. A numerical study of this coupled pedestrian-vehicle delay problem has been presented in a previous paper. Here, we focus on the analytical treatment of the problem, which requires us to use a simplified car-following model. Our analytical results for the transition to oscillatory pedestrian and traffic flows and the average waiting times are well supported by numerical evaluations and give a detailed picture of the collective dynamics emerging when pedestrians try to cross a road. The mathematical expressions allow one to identify the dependence on model parameters such as the vehicle or pedestrian arrival rate, and the safety factor of pedestrian gap acceptance. We also calculate a formula for the vehicle time gap distribution, which corresponds to the departure time contribution of a M/D/1 queue characterized by Poissonian distributed Markovian arrivals, 1 service channel, and deterministic departures. PMID- 16383492 TI - Phase separation driven by surface diffusion: a Monte Carlo study. AB - We propose a kinetic Ising model to study phase separation driven by surface diffusion. This model is referred to as Model S, and consists of the usual Kawasaki spin-exchange kinetics (Model B) in conjunction with a kinetic constraint. We use multispin coding techniques to develop fast algorithms for Monte Carlo simulations of Models B and S. We use these algorithms to study the late stages of pattern dynamics in these systems, and compare properties of the evolution morphologies, e.g., growth laws, domain distribution functions, and spatial and temporal correlation functions. PMID- 16383493 TI - Critical phase in nonconserving zero-range processes and rewiring networks. AB - Zero-range processes, in which particles hop between sites on a lattice, are closely related to rewiring networks, in which rewiring of links between nodes takes place. Both systems exhibit a condensation transition for appropriate choices of the dynamical rules. The transition results in a macroscopically occupied site for zero-range processes and a macroscopically connected node for networks. Criticality, characterized by a scale-free distribution, is obtained only at the transition point. This is in contrast with the widespread scale-free complex networks. Here we propose a generalization of these models whereby criticality is obtained throughout an entire phase, and the scale-free distribution does not depend on any fine-tuned parameter. PMID- 16383494 TI - Systematic analysis of group identification in stock markets. AB - We propose improved methods to identify stock groups using the correlation matrix of stock price changes. By filtering out the market-wide effect and the random noise, we construct the correlation matrix of stock groups in which nontrivial high correlations between stocks are found. Using the filtered correlation matrix, we successfully identify the multiple stock groups without any extra knowledge of the stocks by the optimization of the matrix representation and the percolation approach to the correlation-based network of stocks. These methods drastically reduce the ambiguities while finding stock groups using the eigenvectors of the correlation matrix. PMID- 16383495 TI - Host-parasite models on graphs. AB - The behavior of two interacting populations "hosts" and "parasites" is investigated on Cayley trees and scale-free networks. In the former case analytical and numerical arguments elucidate a phase diagram for the susceptible infected-susceptible model, whose most interesting feature is the absence of a tricritical point as a function of the two independent spreading parameters. For scale-free graphs, the parasite population can be described effectively by its dynamics in a host background. This is shown both by considering the appropriate dynamical equations and by numerical simulations on Barabasi-Albert networks with the major implication that in the thermodynamic limit the critical parasite spreading parameter vanishes. Some implications and generalizations are discussed. PMID- 16383496 TI - Statistical-mechanical iterative algorithms on complex networks. AB - The Ising models have been applied for various problems on information sciences, social sciences, and so on. In many cases, solving these problems corresponds to minimizing the Bethe free energy. To minimize the Bethe free energy, a statistical-mechanical iterative algorithm is often used. We study the statistical-mechanical iterative algorithm on complex networks. To investigate effects of heterogeneous structures on the iterative algorithm, we introduce an iterative algorithm based on information of heterogeneity of complex networks, in which higher-degree nodes are likely to be updated more frequently than lower degree ones. Numerical experiments clarified that the usage of the information of heterogeneity affects the algorithm in Barabasi and Albert networks, but does not influence that in Erdos and Renyi networks. It is revealed that information of the whole system propagates rapidly through such high-degree nodes in the case of Barabasi-Albert's scale-free networks. PMID- 16383497 TI - Exact renormalization group approach to a nonlinear diffusion equation. AB - The exact renormalization group is applied to a nonlinear diffusion equation with a discontinuous diffusion coefficient. The generating functional of the solution for the initial-value problem of nonlinear diffusion equations is first introduced, and next a regularization scheme is presented. It is shown that the renormalization of an action functional in the generating functional leads to an anomalous diffusion exponent in full order of the perturbation series with respect to a nonlinearity. PMID- 16383498 TI - Creation-annihilation processes in the ensemble of constant particle number. AB - We study, in the ensemble of constant particle number, processes in which a cluster of particles is annihilated and particles are created catalytically in active sites. In this ensemble, particles belonging to a cluster of l particles jump to l distinct active sites. As examples of our prescription, we analyze numerically three nonequilibrium systems that annihilate cluster of particles that are identified as conserved versions of the pair annihilation contact model, triplet annihilation contact model, and pair contact process. We show also how to set up the constant particle number ensemble from the constant rate ensemble. PMID- 16383499 TI - Rank distributions of words in correlated symbolic systems and the Zipf law. AB - The binary many-step Markov chain with the step-like memory function is considered as a model for the analysis of rank distributions of words in correlated stochastic symbolic systems. We prove that this distribution obeys the power law with the exponent of the order of unity in the case of rather strong persistent correlations. The Zipf law is shown to be valid for the rank distribution of words with lengths about and shorter than the correlation length in the Markov sequence. A self-similarity in the rank distribution with respect to the decimation procedure is observed. PMID- 16383500 TI - Self-organized Boolean game on networks. AB - A model of a Boolean game with only one free parameter p that denotes the strength of local interaction is proposed wherein each agent acts according to the information obtained from his neighbors in the network, and those in the minority are rewarded. The simulation results indicate that the dynamic of the system is sensitive to network topology, whereby the network of larger degree variance, i.e., the system of greater information heterogeneity, leads to less system profit. The system can self-organize to a stable state and perform better than the random choice game, although only the local information is available to the agents. In addition, in heterogeneity networks, the agents with more information gain more than those with less information for a wide extent of interaction strength p. PMID- 16383501 TI - Mutual selection model for weighted networks. AB - For most networks, the connection between two nodes is the result of their mutual affinity and attachment. In this paper, we propose a mutual selection model to characterize the weighted networks. By introducing a general mechanism of mutual selection, the model can produce power-law distributions of degree, weight, and strength, as confirmed in many real networks. Moreover, we also obtained the nontrivial clustering coefficient C, degree assortativity coefficient r, and degree-strength correlation depending on a single parameter m. These results are supported by present empirical evidence. Studying the degree-dependent average clustering coefficient C(k) and the degree-dependent average nearest neighbors' degree k(nn)(k) also provide us with a better description of the hierarchies and organizational architecture of weighted networks. PMID- 16383502 TI - Analytic calculation of energy transfer and heat flux in a one-dimensional system. AB - In the context of the problem of heat conduction in one-dimensional systems, we present an analytical calculation of the instantaneous energy transfer across a tagged particle in a one-dimensional gas of equal-mass, hard-point particles. From this, we obtain a formula for the steady-state energy flux, and identify and separate the mechanical work and heat conduction contributions to it. The nature of the Fourier law for the model, and the nonlinear dependence of the rate of mechanical work on the stationary drift velocity of the tagged particle, are analyzed and elucidated. PMID- 16383503 TI - Synchronous and asynchronous recursive random scale-free nets. AB - We investigate the differences between scale-free recursive nets constructed by a synchronous, deterministic updating rule (e.g., Apollonian nets), versus an asynchronous, random sequential updating rule (e.g., random Apollonian nets). We show that the dramatic discrepancies observed recently for the degree exponent in these two cases result from a biased choice of the units to be updated sequentially in the asynchronous version. PMID- 16383504 TI - Optimization of random searches on regular lattices. AB - We investigate random searches on isotropic and topologically regular square and triangular lattices with periodic boundary conditions and study the efficiency of search strategies based on a power-law distribution P() approximately (-mu) of step lengths . We consider both destructive searches, in which a target can be visited only once, and nondestructive searches, when a target site is always available for future visits. We discuss (i) the dependence of the search efficiency on the choice of the lattice topology, (ii) the relevance of the periodic boundary conditions, (iii) the behavior of the optimal power-law exponent mu(opt) as a function of target site density, (iv) the differences between destructive and nondestructive environments, and finally (v) how the results for the discrete searches differ from the continuous cases previously studied. PMID- 16383505 TI - Survival probabilities of history-dependent random walks. AB - We analyze the dynamics of random walks with long-term memory (binary chains with long-range correlations) in the presence of an absorbing boundary. An analytically solvable model is presented, in which a dynamical phase transition occurs when the correlation strength parameter mu reaches a critical value mu(c). For strong positive correlations, mu > mu(c), the survival probability is asymptotically finite, whereas for mu < mu(c) it decays as a power law in time (chain length). PMID- 16383506 TI - Sensing the distance to a source of periodic oscillations in a nonlinear chemical medium with the output information coded in frequency of excitation pulses. AB - A spatially distributed excitable chemical medium can collect and process information coded in the propagating pulses of excitation. We consider the problem of distance sensing with the use of a nonlinear chemical medium. We demonstrate that a sensor that can feel the distance separating it from a source of periodic excitations can be constructed by a proper geometrical arrangement of excitable and nonexcitable regions. The sensor returns information about the distance in the frequency of outgoing pulses. The sensor functionality is tested by simulations based on the Rovinsky-Zhabotinsky model. The results are confirmed in experiments performed for a ruthenium-catalyzed Belousov-Zhabotinsky reaction. PMID- 16383507 TI - Asymptotical forms of canonical mappings near separatrix of Hamiltonian systems. AB - Asymptotical behavior of canonical mappings near the separatrix of Hamiltonian systems subjected to time-periodic perturbations is studied. Based on general forms of these mappings [S. S. Abdullaev, Phys. Rev. E 70, 046202 (2004)] it is shown that the Melnikov-type integrals determining their generating functions can be presented as a sum of regular, R(reg)(h), and oscillatory, R(osc)(h), parts. General asymptotical formulas for R(osc)(h) are derived. The oscillatory parts have zeros at primary resonant values of energy. Conditions are found at which the oscillatory parts, R(osc)(h), can be neglected in the generating functions thus allowing us to obtain simplified mappings depending only the regular parts R(reg)(h). Since the latter are smooth functions of energy h this allows us also to justify the widely used conventional separatrix mapping determined by R(reg)(h) at the separatrix h = 0. A theory is illustrated for a specific example of a Hamiltonian system, a particle dynamics in periodically perturbed double well potential. PMID- 16383508 TI - Control of unstable steady states by time-delayed feedback methods. AB - We show that time-delayed feedback methods, which have successfully been used to control unstable periodic orbits, provide a tool to stabilize unstable steady states. We present an analytical investigation of the feedback scheme using the Lambert function and discuss effects of both a low-pass filter included in the control loop and nonzero latency times associated with the generation and injection of the feedback signal. PMID- 16383509 TI - Front propagation and mode-locking in an advection-reaction-diffusion system. AB - Experiments are presented on chemical front propagation in an oscillating chain of vortices in which the mixing of passive impurities is chaotic. The excitable ruthenium-catalyzed Belousov-Zhabotinsky reaction is used in these studies. Velocities of the propagating fronts are measured as a function of the frequency and amplitude of external forcing. Mode locking is observed where the front propagates an integer number of vortices in an integer number of drive periods. Arnol'd tongues are mapped out for two of the locking regimes. These two tongues are shown to form a region of overlap where the velocity of the propagating front switches erratically between two locked values. The experimental results agree with numerical predictions of mode locking in a simplified model of the flow. PMID- 16383510 TI - General approach to the quantum kicked particle in a magnetic field: quantum antiresonance transition. AB - The quantum kicked particle in a magnetic field is studied in a weak-chaos regime under realistic conditions, i.e., for general values of the conserved coordinate x(c) of the cyclotron orbit center. The system exhibits spectral structures ["Hofstadter butterflies" (HBs)] and quantum diffusion depending sensitively on x(c). Most significant changes take place when x(c) approaches the value at which quantum antiresonance (exactly periodic recurrences) can occur: the HB essentially "doubles" and the quantum-diffusion coefficient D(x(c)) is strongly reduced. An explanation of these phenomena, including an approximate formula for D(x(c)) in a class of wave packets, is given on the basis of an effective Hamiltonian which is derived as a power expansion in a small parameter. The global quantum diffusion of a two-dimensional wave packet for all x(c) is briefly considered. PMID- 16383511 TI - Removal of a pinned spiral by generating target waves with a localized stimulus. AB - Pinning of spiral waves by defects in cardiac muscle may cause permanent tachycardia. We numerically study the removal of a pinned spiral by a localized stimulus at the boundary of a two-dimensional excitable medium. It is shown that target waves may be generated by an external local force, and then the target waves will interact with the pinned spiral. When the external force is appropriately chosen, the generated target waves may suppress the pinned spiral, and the system is finally dominated by the target waves. PMID- 16383512 TI - Periodic-orbit theory of universality in quantum chaos. AB - We argue semiclassically, on the basis of Gutzwiller's periodic-orbit theory, that full classical chaos is paralleled by quantum energy spectra with universal spectral statistics, in agreement with random-matrix theory. For dynamics from all three Wigner-Dyson symmetry classes, we calculate the small-time spectral form factor K(tau) as power series in the time tau. Each term tau(n) of that series is provided by specific families of pairs of periodic orbits. The contributing pairs are classified in terms of close self-encounters in phase space. The frequency of occurrence of self-encounters is calculated by invoking ergodicity. Combinatorial rules for building pairs involve nontrivial properties of permutations. We show our series to be equivalent to perturbative implementations of the nonlinear sigma models for the Wigner-Dyson ensembles of random matrices and for disordered systems; our families of orbit pairs have a one-to-one relationship with Feynman diagrams known from the sigma model. PMID- 16383513 TI - Semiclassical analysis of real atomic spectra beyond Gutzwiller's approximation. AB - Real atomic systems, like the hydrogen atom in a magnetic field or the helium atom, whose classical dynamics are chaotic, generally present both discrete and continuous symmetries. In this paper, we explain how these properties must be taken into account in order to obtain the proper (i.e., symmetry projected) h expansion of semiclassical expressions like the Gutzwiller trace formula. In the case of the hydrogen atom in a magnetic field, we shed light on the excellent agreement between present theory and exact quantum results. PMID- 16383514 TI - Regularization of dynamics in an ensemble of nondiffusively coupled chaotic elements. AB - We investigate the dynamics in an ensemble of chaotic elements with nondiffusive coupling. First, we analyze the case of global coupling. The type of coupling we consider leads to the suppression of oscillations in the whole ensemble at a high coupling strength. A distinct feature of this transition from high-dimensional chaos at a low coupling strength to the stationary state is that there is no partially ordered phase characterized by a large number of coexisting synchronized clusters. A two-cluster mode emerges abruptly, replacing the asynchronous mode. We focus on the influence of connectivity on the dynamics in the two-cluster modes and their domains of existence. We introduce a parameter that characterizes the connectivity: the range of coupling. Our computational and analytical results indicate that the most significant changes in the dynamics occur in the case of local coupling, when extra connections are added. By contrast, if the range of coupling is high, even substantial changes in this range have a small influence on the dynamics. PMID- 16383515 TI - Static, oscillating modulus, and moving pulses in the one-dimensional quintic complex Ginzburg-Landau equation: an analytical approach. AB - By means of a matching approach we study analytically the appearance of static and oscillating-modulus pulses in the one-dimensional quintic complex Ginzburg Landau equation without nonlinear gradient terms. When considering nonlinear gradient terms the method enables us to calculate the velocities of the stable and unstable moving pulses. We focus on this equation since it represents a prototype envelope equation associated with the onset of an oscillatory instability near a weakly inverted bifurcation. The results obtained using the analytic approximation scheme are in good agreement with direct numerical simulations. The method is also useful in studying other localized structures like holes. PMID- 16383516 TI - Thermodynamic limit of the first-order phase transition in the Kuramoto model. AB - In the Kuramoto model, a uniform distribution of the natural frequencies leads to a first-order (i.e., discontinuous) phase transition from incoherence to synchronization, at the critical coupling parameter K(c). We obtain the asymptotic dependence of the order parameter above criticality: r-r(c)alpha(K - K(c))(2/3). For a finite population, we demonstrate that the population size N may be included into a self-consistency equation relating r and K in the synchronized state. We analyze the convergence to the thermodynamic limit of two alternative schemes to set the natural frequencies. Other frequency distributions different from the uniform one are also considered. PMID- 16383517 TI - Complete and flexible replacement of chaotic uncertainty with transmitted information. AB - Natural chaos can be described as an information source emitting symbolic sequences with positive entropy. We use two algorithmic techniques from data compression in a nonstandard way along with a control scheme to replace the natural uncertainty in chaotic systems with an arbitrary digital message. Unlike previous targeting-based control, the controlled, deterministic, transmission appears statistically identical to natural chaos, with a message modulated on it at the intrinsic Kolmogorov-Sinai information generation rate of the chaotic oscillator. Thus, chaotic communication by targeting need not consume any additional channel capacity beyond that required by the message itself and the message-bearing signal may appear identical to the uncontrolled oscillator. We also demonstrate control and data transmission at the channel capacity of the oscillator, the maximum possible data rate compatible with the grammar. PMID- 16383518 TI - Formation of multifractal population patterns from reproductive growth and local resettlement. AB - We consider the general character of the spatial distribution of a population that grows through reproduction and subsequent local resettlement of new population members. We present several simple one- and two-dimensional point placement models to illustrate possible generic behavior of these distributions. We show, numerically and analytically, that these models all lead to multifractal spatial distributions of population. Additionally, we make qualitative links between our models and the example of the Earth at Night image, showing the Earth's nighttime man-made light as seen from space. The Earth at Night data suffer from saturation of the sensing photodetectors at high brightness ("clipping"), and we account for how this influences the determined dimension spectrum of the light intensity distribution. PMID- 16383519 TI - Front explosion in a periodically forced surface reaction. AB - Resonantly forced oscillatory reaction-diffusion systems can exhibit fronts with complicated interfacial structure separating phase-locked homogeneous states. For values of the forcing amplitude below a critical value the front "explodes" and the width of the interfacial zone grows without bound. Such front explosion phenomena are investigated for a realistic model of catalytic CO oxidation on a Pt(110) surface in the 2:1 and 3:1 resonantly forced regimes. In the 2:1 regime, the fronts are stationary and the front explosion leads to a defect-mediated turbulent state. In the 3:1 resonantly forced system, the fronts propagate. The front velocity tends to zero as the front explosion point is reached and the final asymptotic state is a 2:1 resonantly locked labyrinthine pattern. The front dynamics described here should be observable in experiment since the model has been shown to capture essential features of the CO oxidation reaction. PMID- 16383520 TI - Observation of spiral pattern and spiral defect chaos in dielectric barrier discharge in argon/air at atmospheric pressure. AB - A rich variety of spiral patterns such as single-armed spiral, dipole spirals, target pattern, multiarmed spiral, and spiral defect chaos state have been observed in ac-driven atmospheric pressure gas discharge. The confined and free boundary conditions are defined by means of whether there is a sidewall in the discharge domain or not, respectively. In the free boundary condition, the spiral pattern arises when the stripe pattern undergoes core instability or notching instability. In the confined boundary condition, the spiral pattern is formed by sidewall forcing. The spiral drifts upward in the free boundary condition and meanders in the confined boundary condition. The topological charge of the spiral pattern can be changed when the spiral interacts with the dislocations. The spiral wavelength (average distance between two consecutive rolls) is a function of gas composition and decreases rapidly with increase of air concentration in discharge gas. PMID- 16383521 TI - Dynamics of traveling waves under spatiotemporal forcing. AB - We study dynamics of traveling waves under spatiotemporal forcing in nonequilibrium systems. Based on the model equations where phase separation and chemical reactions take place simultaneously so that traveling waves are formed in a self-organized manner, we apply a space-time dependent external force. Entrainment and modulation of traveling waves are investigated numerically in one dimension. We develop a theoretical analysis to understand the dynamics obtained. PMID- 16383522 TI - Detecting nonlinearity in structural systems using the transfer entropy. AB - The transfer entropy was recently proposed as a means of exploring coupling in dynamical systems. Transfer entropy is an information theoretic that quantifies the degree to which one dynamical process affects the transition probabilities (dynamics) of another. Here we demonstrate how this metric may be utilized to detect the presence of nonlinearity in a system. Using the method of surrogate data, the transfer entropy computed at various lag times are compared to values computed from linearized surrogates. The transfer entropy is shown to be a more sensitive indicator of nonlinearity than is the mutual information for both simulated and experimental data. This technique is particularly applicable to the field of structural health monitoring, where damage is often equated with the presence of a nonlinearity in an otherwise linear system. PMID- 16383523 TI - Approach to predictability via anticipated synchronization. AB - Predictability of chaotic systems is limited, in addition to the precision of the knowledge of the initial conditions, by the error of the models used to extract the nonlinear dynamics from the time series. In this paper, we analyze the predictions obtained from the anticipated synchronization scheme using a chain of slave neural network approximate replicas of the master system. We compare the maximum prediction horizons obtained with those attainable using standard prediction techniques. PMID- 16383524 TI - Irreversibility with quantum trajectories. AB - Irreversibility is an important issue for many quantum processes. Loschmidt echoes, originally introduced as a way to gauge sensitivity to perturbations in quantum mechanics, have turned out to be a useful tool for its investigation. Following the philosophy supporting this idea, and using quantum trajectories as defined in the causal interpretation of quantum mechanics due to Bohm, we introduce in this paper a more informative alternative measure for irreversibility. The method is applied to the Bunimovich stadium billiard, a paradigmatic example of chaotic system, that constitutes an excellent model for mesoscopic devices. PMID- 16383525 TI - Visualization of coupling in time series by order recurrence plots. AB - We introduce a method to visualize dependencies between two time series by applying the concept of cross recurrence plots to the local ordinal structure. We derive a measure of the coupling strength which is robust against observational noise, nonlinear distortion of the amplitude, and low-frequency trends. Connections to the instantaneous phase and the determination of phase coupling of two coupled Rossler systems in the standard and funnel regimes are shown. An application to electroencephalogram data demonstrates that the method is robust with respect to artifacts. PMID- 16383526 TI - Model for line defects in complex-oscillatory spiral waves. AB - Spiral waves in period-doubled and other complex-oscillatory media possess line defects across which the phase of the oscillation changes by multiples of 2pi. For such systems, the concept of a splay state, introduced for coupled oscillator systems, is generalized to an Archimedean spiral splay field. In this splay field a spiral wave in a two dimensional space is considered to be a special splay state where spatial points having identical phase space orbits take phases determined by the Archimedean spiral on which they lie. Using the Archimedean spiral splay field, an equation that determines the shape of the line defect is derived. PMID- 16383527 TI - Shell-to-shell energy transfer in magnetohydrodynamics. I. Steady state turbulence. AB - We investigate the transfer of energy from large scales to small scales in fully developed forced three-dimensional magnetohydrodynamics (MHD) turbulence by analyzing the results of direct numerical simulations in the absence of an externally imposed uniform magnetic field. Our results show that the transfer of kinetic energy from large scales to kinetic energy at smaller scales and the transfer of magnetic energy from large scales to magnetic energy at smaller scales are local, as is also found in the case of neutral fluids and in a way that is compatible with the Kolmogorov theory of turbulence. However, the transfer of energy from the velocity field to the magnetic field is a highly nonlocal process in Fourier space. Energy from the velocity field at large scales can be transferred directly into small-scale magnetic fields without the participation of intermediate scales. Some implications of our results to MHD turbulence modeling are also discussed. PMID- 16383528 TI - Shell-to-shell energy transfer in magnetohydrodynamics. II. Kinematic dynamo. AB - We study the transfer of energy between different scales for forced three dimensional magnetohydrodynamics turbulent flows in the kinematic dynamo regime. Two different forces are examined: a nonhelical Taylor-Green flow with magnetic Prandtl number P(M) = 0.4 and a helical ABC flow with P(M) = 1. This analysis allows us to examine which scales of the velocity flow are responsible for dynamo action and identify which scales of the magnetic field receive energy directly from the velocity field and which scales receive magnetic energy through the cascade of the magnetic field from large to small scales. Our results show that the turbulent velocity fluctuations in the inertial range are responsible for the magnetic field amplification at small scales (small-scale dynamo) while the large scale field is amplified mostly due to the large-scale flow. A direct cascade of the magnetic field energy from large to small scales is also presented and is a complementary mechanism for the increase of the magnetic field at small scales. The input of energy from the inertial range velocity field into the small magnetic scales dominates over the energy cascade up to the wave number where the magnetic energy spectrum peaks. At even smaller scales, most of the magnetic energy input is from the cascading process. PMID- 16383529 TI - Deposit growth in the wetting of an angular region with uniform evaporation. AB - Solvent loss due to evaporation in a drying drop can drive capillary flows and solute migration. The flow is controlled by the evaporation profile and the geometry of the drop. We predict the flow and solute migration near a sharp corner of the perimeter under the conditions of uniform evaporation. This extends the study of Popov and Witten [Phys. Rev. E 68, 036306 (2003)], which considered a singular evaporation profile, characteristic of a dry surrounding surface. We find the rate of the deposit growth along contact lines in early and intermediate time regimes. Compared to the dry-surface evaporation profile of Popov and Witten [Phys. Rev. E 68, 036306 (2003)], uniform evaporation yields more singular deposition in the early time regime, and nearly uniform deposition profile is obtained for a wide range of opening angles in the intermediate time regime. Uniform evaporation also shows a more pronounced contrast between acute opening angles and obtuse opening angles. PMID- 16383530 TI - Cross-property correlations and permeability estimation in sandstone. AB - We computationally investigate cross-property correlations linking fluid permeability to conductive properties in sedimentary rock for a number of pore size parameters based on three-dimensional digitized rock images. In particular, we focus on correlations based on the pore volume-to-surface-area ratio (V(p)/S), a critical channel diameter (c) associated with mercury porosimetry measurements, length scales associated with the nuclear magnetic resonance relaxation time T2, as well as the mean survival time (tau). Differences between the length scales are discussed. All these correlations yield good agreement with our simulations, but permeability estimates based on the critical diameter (c) are found to be most reliable. PMID- 16383531 TI - Numerical and experimental study of dissociation in an air-water single-bubble sonoluminescence system. AB - We performed a comprehensive numerical and experimental analysis of dissociation effects in an air bubble in water acoustically levitated in a spherical resonator. Our numerical approach is based on suitable models for the different effects considered. We compared model predictions with experimental results obtained in our laboratory in the whole phase parameter space, for acoustic pressures from the bubble dissolution limit up to bubble extinction. The effects were taken into account simultaneously to consider the transition from nonsonoluminescence to sonoluminescence bubbles. The model includes (1) inside the bubble, transient and spatially nonuniform heat transfer using a collocation points method, dissociation of O2 and N2, and mass diffusion of vapor in the noncondensable gases; (2) at the bubble interface, nonequilibrium evaporation and condensation of water and a temperature jump due to the accommodation coefficient; (3) in the liquid, transient and spatially nonuniform heat transfer using a collocation points method, and mass diffusion of the gas in the liquid. The model is completed with a Rayleigh-Plesset equation with liquid compressible terms and vapor mass transfer. We computed the boundary for the shape instability based on the temporal evolution of the computed radius. The model is valid for an arbitrary number of dissociable gases dissolved in the liquid. We also obtained absolute measurements for R(t) using two photodetectors and Mie scattering calculations. The robust technique used allows the estimation of experimental results of absolute R0 and P(a). The technique is based on identifying the bubble dissolution limit coincident with the parametric instability in (P(a),R0) parameter space. We take advantage of the fact that this point can be determined experimentally with high precision and replicability. We computed the equilibrium concentration of the different gaseous species and water vapor during collapse as a function of P(a) and R0. The model obtains from first principles the result that in sonoluminescence the bubble is practically 100% argon for air dissolved in water. Therefore, the dissociation reactions in air bubbles must be taken into account for quantitative computations of maximum temperatures. The agreement found between the numerical and experimental data is very good in the whole parameter space explored. We do not fit any parameter in the model. We believe that we capture all the relevant physics with the model. PMID- 16383532 TI - Instability of a planar expansion wave. AB - An expansion wave is produced when an incident shock wave interacts with a surface separating a fluid from a vacuum. Such an interaction starts the feedout process that transfers perturbations from the rippled inner (rear) to the outer (front) surface of a target in inertial confinement fusion. Being essentially a standing sonic wave superimposed on a centered expansion wave, a rippled expansion wave in an ideal gas, like a rippled shock wave, typically produces decaying oscillations of all fluid variables. Its behavior, however, is different at large and small values of the adiabatic exponent gamma. At gamma > 3, the mass modulation amplitude delta(m) in a rippled expansion wave exhibits a power-law growth with time alpha(t)beta, where beta = (gamma - 3)/(gamma - 1). This is the only example of a hydrodynamic instability whose law of growth, dependent on the equation of state, is expressed in a closed analytical form. The growth is shown to be driven by a physical mechanism similar to that of a classical Richtmyer Meshkov instability. In the opposite extreme gamma - 1 << 1, delta(m) exhibits oscillatory growth, approximately linear with time, until it reaches its peak value approximately (gamma - 1)(-1/2), and then starts to decrease. The mechanism driving the growth is the same as that of Vishniac's instability of a blast wave in a gas with low . Exact analytical expressions for the growth rates are derived for both cases and favorably compared to hydrodynamic simulation results. PMID- 16383533 TI - Theory and modeling of accelerating flames in tubes. AB - The analytical theory of premixed laminar flames accelerating in tubes is developed, which is an important part of the fundamental problem of flame transition to detonation. According to the theory, flames with realistically large density drop at the front accelerate exponentially from a closed end of a tube with nonslip at the walls. The acceleration is unlimited in time; it may go on until flame triggers detonation. The analytical formulas for the acceleration rate, for the flame shape and the velocity profile in the flow pushed by the flame are obtained. The theory is validated by extensive numerical simulations. The numerical simulations are performed for the complete set of hydrodynamic combustion equations including thermal conduction, viscosity, diffusion, and chemical kinetics. The theoretical predictions are in a good agreement with the numerical results. It is also shown how the developed theory can be used to understand acceleration of turbulent flames. PMID- 16383534 TI - Dynamical systems model of entrainment due to coherent structures. AB - The lifetime of a turbulent shear flow is inversely related to the amount of surrounding fluid it entrains. A dynamical systems model of such a flow, dominated by coherent structures, was recently proposed [R. Govindarajan, Phys. Rev. Lett. 88, 134503 (2002)], where an axisymmetric heated jet was modelled by a pair of leap-frogging vortex rings. In the present paper we study the equivalent two-dimensional flow, namely, two pairs of corotating point vortices, of opposite sign. We prove that the system is nonintegrable for any finite separation between the pairs, and show that entrainment is a natural consequence of chaotic advection. The effect of core diffusion due to viscosity is studied, and it is shown that the entrainment rate is reduced with increasing viscosity, by a reduction in leap-frogging frequency. PMID- 16383535 TI - Sonochemical effects on single-bubble sonoluminescence. AB - A refined hydrochemical model for single-bubble sonoluminescence (SBSL) is presented. The processes of water vapor evaporation and condensation, mass diffusion, and chemical reactions are taken into account. Numerical simulations of Xe-, Ar- and He-filled bubbles are carried out. The results show that the trapped water vapor in conjunction with its endothermic chemical reactions significantly reduces the temperature within the bubble so that the degrees of ionization are generally very low. The chemical radicals generated from water vapor are shown to play an increasingly important role in the light emission from Xe to He bubbles. Light spectra and pulses are then computed from an optically thin model. It is found that the resulting spectrum intensities are too small and the pulse widths are too short to fit to recent experimental results within stable SBSL range. Addition of a finite-size blackbody core to the optically thin model improves the fitting. Suggestions on how to reconcile the conflict are given. PMID- 16383536 TI - Buoyancy-drag mix model obtained by multifluid interpenetration equations. AB - In this paper, a buoyancy drag equation for describing the motion of the edges of a mixing zone driven by Rayleigh-Taylor or Richtmyer-Meshkov instabilities is derived from the multifluid interpenetration mix model equations of Scannapieco and Cheng [Phys. Letters A 299, 49 (2002)]. This derivation provides a physics foundation for a large class of phenomenological buoyancy-drag mix models and also establishes a physical connection between the microscopic collision frequency and the macroscopic fluid drag coefficient. The predicted values for model parameter a(ss') in the multifluid interpenetration mix model, from the Rocket-Rig experiments, is in the range of 0.043-0.125 depending upon the Atwood number. The results are also in good agreement with inertial confinement fusion capsule implosions. PMID- 16383537 TI - Charge transport scaling in turbulent electroconvection. AB - We describe a local-power-law scaling theory for the mean dimensionless electric current Nu in turbulent electroconvection. The experimental system consists of a weakly conducting, submicron-thick liquid-crystal film supported in the annulus between concentric circular electrodes. It is driven into electroconvection by an applied voltage between its inner and outer edges. At sufficiently large voltage differences, the flow is unsteady and electric charge is turbulently transported between the electrodes. Our theoretical development, which closely parallels the Grossmann-Lohse model for turbulent thermal convection, predicts the local-power law Nu approximately F(gamma)R(gamma)P(delta). R and P are dimensionless numbers that are similar to the Rayleigh and Prandtl numbers of thermal convection, respectively. The dimensionless function F(gamma), which is specified by the model, describes the dependence of Nu on the aspect ratio gamma. We find that measurements of Nu are consistent with the theoretical model. PMID- 16383538 TI - Lattice Boltzmann method for non-Newtonian (power-law) fluids. AB - We study an ad hoc extension of the lattice Boltzmann method that allows the simulation of non-Newtonian fluids described by generalized Newtonian models. We extensively test the accuracy of the method for the case of shear-thinning and shear-thickening truncated power-law fluids in the parallel plate geometry, and show that the relative error compared to analytical solutions decays approximately linear with the lattice resolution. Finally, we also tested the method in the reentrant-flow geometry, in which the shear rate is no longer a scalar and the presence of two singular points requires high accuracy in order to obtain satisfactory resolution in the local stress near these points. In this geometry, we also found excellent agreement with the solutions obtained by standard finite-element methods, and the agreement improves with higher lattice resolution. PMID- 16383539 TI - Direct acceleration of solid-density plasma bunch by ultraintense laser. AB - The interaction of a petawatt laser with a small solid-density plasma bunch is studied by particle-in-cell simulation. It is shown that when irradiated by a laser of intensity >10(21) W/cm2, a dense plasma bunch of micrometer size can be efficiently accelerated. The kinetic energy of the ions in the high-density region of the plasma bunch can exceed ten MeV at a density in the 10(23)-cm(-3) level. Having a flux density orders of magnitude higher than that of the traditional charged-particle pulses, the laser-accelerated plasma bunch can have a wide range of applications. In particular, such a dense energetic plasma bunch impinging on the compressed fuel in inertial fusion can significantly enhance the nuclear-reaction cross section and is thus a promising alternative for fast ignition. PMID- 16383540 TI - Equation of state for partially ionized carbon at high temperatures. AB - Equation of state for partially ionized carbon at temperatures T approximately > or = 10(5) K is calculated in a wide range of densities, using the method of free energy minimization in the framework of the chemical picture of plasmas. The free energy model includes the internal partition functions of bound species. The latter are calculated by a self-consistent treatment of each ionization stage in the plasma environment taking into account pressure ionization. The long-range Coulomb interactions between ions and screening of the ions by free electrons are included using our previously published analytical model. PMID- 16383541 TI - Dust transport in a magnetized radio-frequency discharge under microgravity conditions. AB - Dust is found in plasmas used in industrial applications, such as microelectronics and solar cell manufacturing, in fusion plasmas, where it is usually the result of plasma-wall interactions, and in plasmas in space, such as planetary atmospheres, cometary tails, planetary rings, interstellar molecular clouds, and star and planet formation regions. In plasma applications, magnetic fields are occasionally used, mainly to confine the plasma. In space, however, magnetic fields are very often present and they may strongly influence the behavior of dusty plasma, for instance in the formation of stars and planets. We extended a fully self-consistent two-dimensional fluid model for radio-frequency discharges by adding a homogeneous axial magnetic field and the effect it has on the transport of plasma species in a low-temperature dusty discharge. We show that the magnetic field has an important effect on the (ambipolar) diffusion of ions and electrons in the bulk of the discharge. This causes an important change in the force balance of the dust particles and in the time scales of the formation of a dust-free void. Finally, we compare the parameters of the modeled discharge with the parameters of a planet formation region around a young stellar object (YSO). We conclude that a magnetic field in both low-temperature rf discharges under micro-gravity conditions and dusty plasmas around YSO's has an important effect on the transport of dust and must be important for the formation of planets and stars. PMID- 16383542 TI - Standard line broadening impact theory for hydrogen including penetrating collisions. AB - In recent years there has been significant interest in the emission spectra from high-density plasmas, as manifested by a number of experiments. At these high densities short range (small impact parameter) interactions become important and these cannot be adequately handled by the standard theory, whose predictions depend on some cutoffs, necessary to preserve unitarity, the long range approximation, and to ensure the validity of a semiclassical picture. Very recently, as a result of a debate concerning the broadening of isolated ion lines, the importance of penetration of bound electron wave functions by plasma electrons has been realized. By softening the interaction, penetration makes perturbative treatments more valid. The penetration effect has now been included analytically into the standard theory. It turns out that the integrations may be done in closed form in terms of the modified Bessel functions K0 and K1. This work develops the new theory and applies it to experimental measurements. PMID- 16383543 TI - Calculations of Stark-broadened line shapes in phase-conjugate degenerate four wave mixing laser spectroscopy. AB - The Stark-broadened line shapes for phase-conjugate degenerate four-wave mixing (PCDFWM) laser spectroscopy are studied. The line profiles are calculated for high-density (N(e) > 10(21) m(-3) plasma conditions and for different contributions of the Doppler broadening. Calculations are performed in the limit of low laser intensities using the perturbation approach. The theoretical model takes into account the Stark effect together with the ion dynamics, the electron collisions, and the Doppler effect. The resultant PCDFWM spectral profiles are significantly less broadened and less asymmetric than the emission profiles but shifted by similar magnitudes. Moreover, the results of our calculations show the sub-Doppler character of the PCDFWM profiles and also their strong dependence on the geometric configuration of the pump and the probe laser beams. PMID- 16383544 TI - Adiabatic formation of quasibound states of antihydrogen. AB - The classical trajectory of an initially unbound positron within the electric field of an antiproton and a uniform magnetic field is simulated in three dimensions. Several simulations are run incorporating experimental parameters used for antihydrogen production, which has been achieved by two different groups [M. Amoretti, Nature (London) 419, 456 (2002); G. Gabrielse, Phys. Rev. Lett. 89, 213401 (2002)]. The simulations indicate that temporary bound states of antihydrogen can form at positive energies, where the energy of the system is defined to be zero when the positron and antiproton are at rest with infinite separation. Such quasibound states, which form only when the magnetic field is present, are typically smaller than in a dimension perpendicular to the magnetic field. An analytical model is developed for a formation cross section, and it is found that quasibound states may form more frequently than stable Rydberg states. PMID- 16383545 TI - Scaling law of plasma turbulence with nonconservative fluxes. AB - It is shown that in the presence of anisotropic kinetic dissipation existence of the scale invariant power law spectrum of plasma turbulence is possible. The obtained scale invariant spectrum is not associated with the constant flux of any physical quantity. Application of the model to the high frequency part of the solar wind turbulence is discussed. PMID- 16383547 TI - Formulation of nonlinear chromaticity in circular accelerators by canonical perturbation method. AB - The formulation of nonlinear chromaticity in circular accelerators based on the canonical perturbation method is presented. Since the canonical perturbation method directly relates the tune shift to the perturbation Hamiltonian, it greatly simplifies the calculation of the nonlinear chromaticity. The obtained integral representation for nonlinear chromaticity can be systematically extended to higher orders. PMID- 16383546 TI - Calculation of electromagnetic field components for a fundamental Gaussian beam. AB - Using a perturbative method, we carefully calculate seventh- and ninth-order expressions for electromagnetic field components for a fundamental Gaussian beam (i.e., a focused TEM00 mode laser beam) propagating in the medium analogous to vacuum. The interaction of a single electron and a focused laser pulse in vacuum has a promising application in the design of a powerful electron accelerator which could compete with the traditional ones. For this, ninth-order accuracy in the expansion describing the focused laser beam is required. When the vector potential corresponds to polarization along the direction of propagation, the number of electromagnetic field components can be reduced from 5 or 6 to 3. Furthermore, we find two rules obeyed by all orders of the vector potential, which greatly simplify the calculation procedure of the vector potential order by order, and make it possible to investigate the behavior of high orders. PMID- 16383548 TI - Phase conjugation with a single bubble. AB - It is recalled how the nonlinear interaction between a gas bubble and an external extra pressure can yield phase conjugation. Using the Glauber representation, we show that the effect of the latter is formally analogous to that of a pi pulse in nuclear magnetic resonance, so that the acoustic equivalent of spin echoes may be expected in a bubble cloud. An experimental attempt to observe phase conjugation is reported in the single-bubble case. PMID- 16383549 TI - Ponderomotive ratchet in a uniform magnetic field. AB - We show how a ratchet effect, generally used in systems with periodic potentials, can also be practiced on charged particles by an ac field alone, in a background magnetic field near the cyclotron resonance. The effect relies entirely on the spatial inhomogeneity of the high-frequency drive, which produces a deterministic asymmetric ponderomotive barrier for undamped particles. Such a barrier can reflect particles incident from one side while transmitting those incident from the opposite side, hence acting somewhat like a Maxwell demon. The necessary fields are perhaps most easily realized in a plasma, though the effect is more general. PMID- 16383550 TI - Giant photonic Hall effect in magnetophotonic crystals. AB - We have considered a simple, square, two-dimensional (2D) PC built of a magneto optic matrix with square holes. It is shown that using such a magnetophotonic crystal it is possible to deflect a light beam at very large angles by applying a nonzero external magnetic field. The effect is called the giant photonic Hall effect (GPHE) or the magnetic superprism effect. The GPHE is based on magneto optical properties, as is the photonic Hall effect [B. A. van Tiggelen and G. L. J. A. Rikken, in, edited by V. M. Shalaev (Springer-Verlag, Berlin, 2002), p. 275]; however GPHE is not caused by asymmetrical light scattering but rather by the influence of an external magnetic field on the photonic band structure. PMID- 16383551 TI - Resonantly absorbing one-dimensional photonic crystals. AB - A compact theoretical description of the effects of dissipation on the propagation of light waves through a multilayer periodic mirror built from resonant absorbing atoms is presented. Depending on the lattice periodicity, ultranarrow photonic gaps, weak polaritonic gaps, as well as rather atypical gap structures may be observed. Because of the atom's absorption line shape Bloch gap modes may acquire quite a cumbersome structure which is thoroughly studied here or which may even disappear when dissipation becomes sufficiently strong. The same approach well applies also to resonantly absorbing photonic crystals based on excitonic resonances. PMID- 16383552 TI - Two-dimensional treatment of the level shift and decay rate in photonic crystals. AB - We present a comprehensive treatment of the level shift and decay rate of a model line source in a two-dimensional photonic crystal (2D PC) composed of circular cylinders. The quantities in this strictly two-dimensional system are determined by the two-dimensional local density of states (2D LDOS), which we compute using Rayleigh-multipole methods. We extend the critical point analysis that is traditionally applied to the 2D DOS (or decay rate) to the level shift. With this, we unify the crucial quantity for experiment--the 2D LDOS in a finite PC- with the band structure and the 2D DOS, 2D LDOS, and level shift in infinite PC's. Consistent with critical point analysis, large variations in the level shift are associated with large variations in the 2D DOS (and 2D LDOS), corroborating a giant anomalous Lamb shift. The boundary of a finite 2D PC can produce resonances that cause the 2D LDOS in a finite 2D PC to differ markedly from the 2D LDOS in an infinite 2D PC. PMID- 16383553 TI - Influence of dispersion on the resonant interaction between three incoherent waves. AB - We study the influence of group-velocity dispersion (or diffraction) on the coherence properties of the parametric three-wave interaction driven from an incoherent pump wave. We show that, under certain conditions, the incoherent pump may efficiently amplify a signal wave with a high degree of coherence, in contrast with the usual kinetic description of the incoherent three-wave interaction. The group-velocity dispersion is shown to be responsible for a spectral filtering process, in which the coherence of the generated signal increases, as the coherence of the pump wave decreases. As a result, the coherence acquired by the signal in the presence of an incoherent pump, is higher than that acquired in the presence of a fully coherent pump. The mechanism underlying this intriguing result is based on the emergence of a mutual coherence between the incoherent pump and the generated idler wave. We calculate explicitly the degree of mutual coherence between the pump and idler waves and show that the two incoherent waves become completely correlated in the full incoherent regime of interaction. The theory is in quantitative agreement with the numerical simulations. To motivate the experimental confirmation of our theory, we characterize the dispersion properties of an actual quadratic nonlinear optical crystal in which the process of signal coherence enhancement induced by pump incoherence may be studied experimentally. PMID- 16383554 TI - Passive retrieval of Rayleigh waves in disordered elastic media. AB - When averaged over sources or disorder, cross correlation of diffuse fields yields the Green's function between two passive sensors. This technique is applied to elastic ultrasonic waves in an open scattering slab mimicking seismic waves in the Earth's crust. It appears that the Rayleigh wave reconstruction depends on the scattering properties of the elastic slab. Special attention is paid to the specific role of bulk to Rayleigh wave coupling, which may result in unexpected phenomena, such as a persistent time asymmetry in the diffuse regime. PMID- 16383555 TI - Apparent superluminality and the generalized Hartman effect in double-barrier tunneling. AB - Recent papers suggest that tunneling wave packets traverse the region of allowed propagation between two potential barriers with superluminal group velocity and in a time independent of barrier separation. This phenomenon has been termed the "generalized Hartman effect" and extended to multiple barriers. Here we show that this delay time is not a transit time but a cavity lifetime. It does not imply superluminal velocity. Reported experimental verifications of this effect are reinterpreted. PMID- 16383556 TI - Sound-field modeling in architectural acoustics by a transport theory: application to street canyons. AB - The transport theory of sound particles is applied to the sound field modeling in architectural acoustics. A theoretical description is proposed for empty enclosures with complex boundary conditions, including both specular and diffuse reflections. As an example, the model is applied to street canyons. Therefore, an asymptotic approach is proposed to reduce the transport equation to a diffusion equation defined by only one parameter, the diffusion coefficient. This coefficient is a function of the reflection law of the building facades, the ratio of specular and diffuse reflections, as well as the street width. The model is then compared to Monte Carlo simulations of the propagation of sound particles in complex enclosures. As expected by the asymptotic approach, the model is in agreement with numerical results, but mainly for small street width and very diffuse reflections. Finally, a discussion is proposed in the conclusion, on the model's capabilities. PMID- 16383557 TI - Matter-wave solitons in nonlinear optical lattices. AB - We introduce a dynamical model of a Bose-Einstein condensate based on the one dimensional (1D) Gross-Pitaevskii equation (GPE) with a nonlinear optical lattice (NOL), which is represented by the cubic term whose coefficient is periodically modulated in the coordinate. The model describes a situation when the atomic scattering length is spatially modulated, via the optically controlled Feshbach resonance, in an optical lattice created by interference of two laser beams. Relatively narrow solitons supported by the NOL are predicted by means of the variational approximation (VA), and an averaging method is applied to broad solitons. A different feature is a minimum norm (number of atoms), N = N(min), necessary for the existence of solitons. The VA predicts N(min) very accurately. Numerical results are chiefly presented for the NOL with the zero spatial average value of the nonlinearity coefficient. Solitons with values of the amplitude A larger than at N = N(min) are stable. Unstable solitons with smaller, but not too small, A rearrange themselves into persistent breathers. For still smaller A, the soliton slowly decays into radiation without forming a breather. Broad solitons with very small A are practically stable, as their decay is extremely slow. These broad solitons may freely move across the lattice, featuring quasielastic collisions. Narrow solitons, which are strongly pinned to the NOL, can easily form stable complexes. Finally, the weakly unstable low-amplitude solitons are stabilized if a cubic term with a constant coefficient, corresponding to weak attraction, is included in the GPE. PMID- 16383558 TI - Connection between electric and magnetic coherence in free electromagnetic fields. AB - We introduce quantitative measures for the description of the electric and magnetic coherence in a stationary, random electromagnetic field at two points, in a volume, and in the Fourier space. These quantities are applied to free electromagnetic fields, and several theorems regarding the relationship between the two types of coherences in such fields are established. Fields which are statistically homogeneous, and those which, in addition, are statistically isotropic are considered separately. Furthermore, the connection between the electric and magnetic coherence is exemplified for some specific statistically homogeneous fields. PMID- 16383559 TI - Self-similar propagation and compression of chirped self-similar waves in asymmetric twin-core fibers with nonlinear gain. AB - Ultrashort-pulse propagation in asymmetric twin-core fiber amplifiers is studied with the aid of self-similarity analysis of the nonlinear Schrodinger-type equation interacting with a source, variable dispersion, variable Kerr nonlinearity, variable gain or loss, and nonlinear gain. Exact chirped pulses that can propagate self-similarly subject to simple scaling rules of this model have been found. It is reported that the pulse position of these chirped pulses can be precisely piloted by appropriately tailoring the dispersion profile. This fact is profitably exploited to achieve optimal pulse compression of these chirped self-similar solutions. PMID- 16383560 TI - Discrete solitons and vortices on anisotropic lattices. AB - We consider the effects of anisotropy on solitons of various types in two dimensional nonlinear lattices, using the discrete nonlinear Schrodinger equation as a paradigm model. For fundamental solitons, we develop a variational approximation that predicts that broad quasicontinuum solitons are unstable, while their strongly anisotropic counterparts are stable. By means of numerical methods, it is found that, in the general case, the fundamental solitons and simplest on-site-centered vortex solitons ("vortex crosses") feature enhanced or reduced stability areas, depending on the strength of the anisotropy. More surprising is the effect of anisotropy on the so-called "super-symmetric" intersite-centered vortices ("vortex squares"), with the topological charge equal to the square's size : we predict in an analytical form by means of the Lyapunov Schmidt theory, and confirm by numerical results, that arbitrarily weak anisotropy results in dramatic changes in the stability and dynamics in comparison with the degenerate, in this case, isotropic, limit. PMID- 16383561 TI - Static contact angle in lattice Boltzmann models of immiscible fluids. AB - We study numerically the capillary rise between two horizontal plates and in a rectangular tube, using a lattice Boltzmann (LB) method. We derive an equation for the static fluid-solid contact angle as a function of the wetting tendency of the walls and test its validity. We show that the generalized Laplace law with two independent radii of curvature is followed in capillary rise in rectangular tubes. Our method removes the history dependence of the fluid-solid contact angle that had been present in earlier LB schemes. PMID- 16383562 TI - Equation-free dynamic renormalization: self-similarity in multidimensional particle system dynamics. AB - We present an equation-free dynamic renormalization approach to the computational study of coarse-grained, self-similar dynamic behavior in multidimensional particle systems. The approach is aimed at problems for which evolution equations for coarse-scale observables (e.g., particle density) are not explicitly available. Our illustrative example involves Brownian particles in a 2D Couette flow; marginal and conditional inverse cumulative distribution functions (ICDFs) constitute the macroscopic observables of the evolving particle distributions. PMID- 16383563 TI - Polymer density functional approach to efficient evaluation of path integrals. AB - A polymer density functional theory (P-DFT) has been extended to the case of quantum statistics within the framework of Feynman path integrals. We start with the exact P-DFT formalism for an ideal open chain and adapt its efficient numerical solution to the case of a ring. We show that, similarly, the path integral problem can, in principle, be solved exactly by making use of the two particle pair correlation function (2p-PCF) for the ends of an open polymer, half of the original. This way the exact data for one-dimensional quantum harmonic oscillator are reproduced in a wide range of temperatures. The exact solution is not, though, reachable in three dimensions (3D) because of a vast amount of storage required for 2p-PCF. In order to treat closed paths in 3D, we introduce a so-called "open ring" approximation which proves to be rather accurate in the limit of long chains. We also employ a simple self-consistent iteration so as to correctly account for the interparticle interactions. The algorithm is speeded up by taking convolutions with the aid of fast Fourier transforms. We apply this approximate path integral DFT (PI-DFT) method to systems within spherical symmetry: 3D harmonic oscillator, atoms of hydrogen and helium, and ions of He and Li. Our results compare rather well to the known data, while the computational effort (some seconds or minutes) is about 100 times less than that with Monte Carlo simulations. Moreover, the well-known "sign problem" is expected to be considerably reduced within the reported PI-DFT, since it allows for a direct estimate of the corresponding partition functions. PMID- 16383564 TI - Overcoming the slowing down of flat-histogram Monte Carlo simulations: cluster updates and optimized broad-histogram ensembles. AB - We study the performance of Monte Carlo simulations that sample a broad histogram in energy by determining the mean first-passage time to span the entire energy space of d-dimensional ferromagnetic Ising/Potts models. We first show that flat histogram Monte Carlo methods with single-spin flip updates such as the Wang Landau algorithm or the multicanonical method perform suboptimally in comparison to an unbiased Markovian random walk in energy space. For the d = 1, 2, 3 Ising model, the mean first-passage time tau scales with the number of spins N = L(d) as tau proportional N2L(z). The exponent z is found to decrease as the dimensionality d is increased. In the mean-field limit of infinite dimensions we find that z vanishes up to logarithmic corrections. We then demonstrate how the slowdown characterized by z > 0 for finite d can be overcome by two complementary approaches--cluster dynamics in connection with Wang-Landau sampling and the recently developed ensemble optimization technique. Both approaches are found to improve the random walk in energy space so that tau proportional N2 up to logarithmic corrections for the d = 1, 2 Ising model. PMID- 16383565 TI - Energy dissipation measures in three-dimensional disordered porous media. AB - The onset of nonlinear flow was analyzed in three-dimensional random, porous granular systems with 60% porosity using a lattice-Boltzmann model. Quantitative analysis was based on participation numbers built on local kinetic energies and energy dissipation rates computed via nonequilibrium kinetic (viscous stress) tensors. In contrast to the kinetic energy participation number, which characterizes the onset of nonlinearity in terms of a transition from a locally concentrated to a dispersed distribution of kinetic energy densities, the nonequilibrium kinetic tensor participation number characterizes the onset of nonlinearity in terms of a transition from a dispersed to a locally concentrated distribution of energy dissipation densities as the flow rate increases. The transition characterized by the nonequilibrium kinetic tensor participation number occurred over a nearly equal or a narrower range of Reynolds numbers when compared to the transition characterized by the kinetic energy participation number. PMID- 16383566 TI - Monte Carlo charged-particle tracking and energy deposition on a Lagrangian mesh. AB - A Monte Carlo algorithm for alpha particle tracking and energy deposition on a cylindrical computational mesh in a Lagrangian hydrodynamics code used for inertial confinement fusion (ICF) simulations is presented. The straight line approximation is used to follow propagation of "Monte Carlo particles" which represent collections of alpha particles generated from thermonuclear deuterium tritium (DT) reactions. Energy deposition in the plasma is modeled by the continuous slowing down approximation. The scheme addresses various aspects arising in the coupling of Monte Carlo tracking with Lagrangian hydrodynamics; such as non-orthogonal severely distorted mesh cells, particle relocation on the moving mesh and particle relocation after rezoning. A comparison with the flux limited multi-group diffusion transport method is presented for a polar direct drive target design for the National Ignition Facility. Simulations show the Monte Carlo transport method predicts about earlier ignition than predicted by the diffusion method, and generates higher hot spot temperature. Nearly linear speed-up is achieved for multi-processor parallel simulations. PMID- 16383567 TI - Equilibrium calculation of transport coefficients for a fluid-particle model. AB - A recently introduced particle-based model for fluid flow, called stochastic rotation dynamics, can be made Galilean invariant by introducing a random shift of the computational grid before collisions. In this paper, it is shown how the Green-Kubo relations derived previously can be resummed to obtain exact expressions for the collisional contributions to the transport coefficients. It is also shown that the collisional contribution to the microscopic stress tensor is not symmetric, and that this leads to an additional viscosity. The resulting identification of the transport coefficients for the hydrodynamic modes is discussed in detail, and it is shown that this does not impose restrictions on the applicability of the model. The collisional contribution to the thermal conductivity, which becomes important for small mean free path and small average particle number per cell, is also derived. PMID- 16383568 TI - Efficient computation of optical forces with the coupled dipole method. AB - We present computational techniques to compute in an efficient way optical forces on arbitrary nanoobjects using the coupled dipole method. We show how the time of computation can be reduced by several orders of magnitude with the help of fast Fourier-transform techniques. We also discuss the influence of different formulations of the electric polarizability of a small scatterer on the accuracy and robustness of the computation of optical forces. PMID- 16383569 TI - Scale-free fuse network and its robustness. AB - The robustness and reliability of scale-free networks are tested as a fuse network. The idea is to examine the robustness of a scale-free network when links are irreversibly removed after failing. Due to inherent characteristics of the fuse network model, the sequence of links removal is deterministic and conditioned to fuse tolerance and connectivity of its ends. It is a different situation from classical robustness analysis of complex networks, when they are usually tested under random fails and deliberate attacks of nodes. The use of this system to study the fracture of elastic material brought some interesting results. PMID- 16383570 TI - Optimal phase-space projection for noise reduction. AB - In this communication we will reexamine the widely studied technique of phase space projection. By imposing a time-domain constraint on the residual noise, we deduce a more general version of the optimal projector, which includes those appearing in previous literature as subcases but does not assume the independence between the clean signal and the noise. As an application, we will apply this technique for noise reduction. Numerical results show that our algorithm has succeeded in augmenting the signal-to-noise ratio for simulated data from the Rossler system and experimental speech record. PMID- 16383571 TI - Least-squares finite-element scheme for the lattice Boltzmann method on an unstructured mesh. AB - A numerical model of the lattice Boltzmann method (LBM) utilizing least-squares finite-element method in space and the Crank-Nicolson method in time is developed. This method is able to solve fluid flow in domains that contain complex or irregular geometric boundaries by using the flexibility and numerical stability of a finite-element method, while employing accurate least-squares optimization. Fourth-order accuracy in space and second-order accuracy in time are derived for a pure advection equation on a uniform mesh; while high stability is implied from a von Neumann linearized stability analysis. Implemented on unstructured mesh through an innovative element-by-element approach, the proposed method requires fewer grid points and less memory compared to traditional LBM. Accurate numerical results are presented through two-dimensional incompressible Poiseuille flow, Couette flow, and flow past a circular cylinder. Finally, the proposed method is applied to estimate the permeability of a randomly generated porous media, which further demonstrates its inherent geometric flexibility. PMID- 16383572 TI - Complexity of time series associated to dynamical systems inferred from independent component analysis. AB - A not trivial problem for every experimental time series associated to a natural system is to individuate the significant variables to describe the dynamics, i.e., the effective degrees of freedom. The application of independent component analysis (ICA) has provided interesting results in this direction, e.g., in the seismological and atmospheric field. Since all natural phenomena can be represented by dynamical systems, our aim is to check the performance of ICA in this general context to avoid ambiguities when investigating an unknown experimental system. We show many examples, representing linear, nonlinear, and stochastic processes, in which ICA seems to be an efficacious preanalysis able to give information about the complexity of the dynamics. PMID- 16383573 TI - Numerical method for interaction between multiparticle and complex structures. AB - We propose a numerical method for dealing with interactions between multiple particles and complex structures. In the method, the structures are represented on a grid by using the level set method. The interactions of particles and structures are calculated by a method based on the discrete element method. The method can treat the interaction between multiparticle and complex structures robustly. PMID- 16383574 TI - Modular synchronization in complex networks. AB - We study the synchronization transition (ST) of a modified Kuramoto model on two different types of modular complex networks. It is found that the ST depends on the type of intermodular connections. For the network with decentralized (centralized) intermodular connections, the ST occurs at finite coupling constant (behaves abnormally). Such distinct features are found in the yeast protein interaction network and the Internet, respectively. Moreover, by applying the finite-size scaling analysis to an artificial network with decentralized intermodular connections, we obtain the exponent associated with the order parameter of the ST to be beta approximately 1 different from beta(MF) approximately 1/2 obtained from the scale-free network with the same degree distribution but the absence of modular structure, corresponding to the mean field value. PMID- 16383575 TI - Convergence of continuous-time quantum walks on the line. AB - The position density of a "particle" performing a continuous-time quantum walk on the integer lattice, viewed on length scales inversely proportional to the time , converges (as tends to infinity) to a probability distribution that depends on the initial state of the particle. This convergence behavior has recently been demonstrated for the simplest continuous-time random walk [N. Konno, Phys. Rev. E 72, 026113 (2005)]. In this Brief Report, we use a different technique to establish the same convergence for a very large class of continuous-time quantum walks, and we identify the limit distribution in the general case. PMID- 16383576 TI - Equivalence between maximum entropy principle and enforcing dU = TdS. AB - We study here a hitherto unexplored microscopic connection between the well-known thermodynamical relation dU = TdS and Jaynes' maximum entropy principle (MaxEnt) for determining probability distributions for the canonical ensemble. PMID- 16383577 TI - Agent preferences and the topology of networks. AB - In this paper, a different framework to study weighed networks is introduced. The idea behind this methodology is to consider that each node of the network is an agent that desires to satisfy his/her preferences in an economic sense. Moreover, the formation of a link between two agents depends on the benefits and costs associated with this link. Therefore, an edge between two given nodes will arise only if the tradeoff between satisfaction and cost for building it is jointly positive. Using a computational framework, I intend to show that depending on the agents' combination of benefits and costs, some very well known networks can naturally arise. PMID- 16383578 TI - Fluctuation-dissipation relations outside the linear response regime in a two dimensional driven lattice gas along the direction transverse to the driving force. AB - We performed numerical experiments on a two-dimensional driven lattice gas, which constitutes a simple stochastic nonequilibrium many-body model. In this model, focusing on the behavior along the direction transverse to the external driving force, we numerically measure transport coefficients and dynamical fluctuations outside the linear response regime far from equilibrium. Using these quantities, we find the validity of the Einstein relation, the Green-Kubo relation and the fluctuation-response relation. PMID- 16383579 TI - Surface partition of large clusters. AB - The surface partition of large clusters is derived analytically for a simple statistical model by using the Laplace-Fourier transformation method. In the limit of small amplitude deformations, a suggested "hills and dales model" reproduces the leading term of the Fisher result for the surface entropy to within a few percent. The model also gives the degeneracy prefactor of large clusters. The surface partition of finite clusters is also discussed. PMID- 16383580 TI - Phase diagrams for an evolutionary prisoner's dilemma game on two-dimensional lattices. AB - The effects of payoffs and noise on the maintenance of cooperative behavior are studied in an evolutionary prisoner's dilemma game with players located on the sites of different two-dimensional lattices. This system exhibits a phase transition from a mixed state of cooperators and defectors to a homogeneous one where only the defectors remain alive. Using Monte Carlo simulations and the generalized mean-field approximations we have determined the phase boundaries (critical points) separating the two phases on the plane of the temperature (noise) and temptation to choose defection. In the zero temperature limit the cooperation can be sustained only for those connectivity structures where three site clique percolation occurs. PMID- 16383581 TI - Potential flow inside an evaporating cylindrical line. AB - An analytical solution to the problem of potential flow inside an evaporating line is obtained. The line is shaped as a half-cylinder lying on a substrate, and evaporates with either pinned or depinned contact lines. The solution is provided through the technique of separation of variables in the velocity potential and stream function formulations. Based on the flow field calculations, it is estimated that the coffee-stain phenomenon should be expected even for uniform evaporation flux throughout the cylindrical surface, provided that the contact lines remain anchored. A simple expression for the velocity potential is also suggested, which reproduces the local velocity vector with excellent accuracy. The vertically averaged velocity is calculated also for other contact line values, revealing for any value an outward liquid flow for pinned lines as opposed to inward flow for depinned lines. PMID- 16383582 TI - Photonic band gap effect and structural color from silver nanoparticle gelatin emulsion. AB - We have fabricated planar structures of silver nanoparticles in monochromatic gelatin emulsion with a continuous spacing ranging from 0.15-0.40 micron using a two-beam interference of a single laser source. Our planar holograms display a colorful "rainbow" pattern and photonic bandgaps covering the visible and IR ranges. We model the planar silver nanoparticle-gelatin composite system using an effective medium approach and good agreement is obtained between theory and experiment. PMID- 16383583 TI - Strongly focused polarized light pulse. AB - We investigate theoretically the electric field of a focused light pulse carrying an inhomogeneous polarization distribution. It is found that the spectra of the polarization components are in general different, thus leading to a spatial spectral distribution that differs from the scalar case. PMID- 16383584 TI - Comment on "Role of liquid compressional viscosity in the dynamics of a sonoluminescing bubble". AB - Moshaii and Bonabi found a new viscous term to the traditional bubble boundary equation and declared it greatly affects the motion of the sonoluminescing bubble. However, the simple analysis and recomputation show it is incorrect. PMID- 16383586 TI - Light scattering enhancement in an aqueous solution of spermine-induced fractal aggregate composite. AB - We performed an experimental investigation on the scattering and extinction properties of an aqueous solution of spermine-induced fractal J aggregates, constituted by porphyrin molecules. In analogy with nanostructured particle composites, the dipolar moments of different porphyrins, not belonging to the aggregates J, are coupled and generate a strong light scattering enhancement. Both the enhanced scattered intensity and the imaginary part of the polarizability of the system obey scaling laws with the optical spectral dimension d0=0.3. PMID- 16383587 TI - Transport on randomly evolving trees. AB - The time process of transport on randomly evolving trees is investigated. By introducing the notions of living and dead nodes, a model of random tree evolution is constructed which describes the spreading in time of objects corresponding to nodes. It is assumed that at t=0 the tree consists of a single living node (root), from which the evolution may begin. At a certain time instant tau> or =0, the root produces v> or =0 living nodes connected by lines to the root which becomes dead at the moment of the offspring production. In the evolution process each of the new living nodes evolves further like a root independently of the others. By using the methods of the age-dependent branching processes we derive the joint distribution function of the numbers of living and dead nodes, and determine the correlation between these node numbers as a function of time. It is proved that the correlation function converges to square root of 3/2 independently of the distributions of v and tau when q1-->1 and t- >infinity. Also analyzed are the stochastic properties of the end nodes; and the correlation between the numbers of living and dead end nodes is shown to change its character suddenly at the very beginning of the evolution process. The survival probability of random trees is investigated and expressions are derived for this probability. PMID- 16383588 TI - Numerical method for accessing the universal scaling function for a multiparticle discrete time asymmetric exclusion process. AB - In the universality class of the one-dimensional Kardar-Parisi-Zhang (KPZ) surface growth, Derrida and Lebowitz conjectured the universality of not only the scaling exponents, but of an entire scaling function. Since and Derrida and Lebowitz's original publication [Phys. Rev. Lett. 80, 209 (1998)] this universality has been verified for a variety of continuous-time, periodic boundary systems in the KPZ universality class. Here, we present a numerical method for directly examining the entire particle flux of the asymmetric exclusion process (ASEP), thus providing an alternative to more difficult cumulant ratios studies. Using this method, we find that the Derrida-Lebowitz scaling function (DLSF) properly characterizes the large-system-size limit (N--> infinity) of a single-particle discrete time system, even in the case of very small system sizes (N< or =22). This fact allows us to not only verify that the DLSF properly characterizes multiple-particle discrete-time asymmetric exclusion processes, but also provides a way to numerically solve for quantities of interest, such as the particle hopping flux. This method can thus serve to further increase the ease and accessibility of studies involving even more challenging dynamics, such as the open-boundary ASEP. PMID- 16383589 TI - Langevin equations for fluctuating surfaces. AB - Exact Langevin equations are derived for the height fluctuations of surfaces driven by the deposition of material from a molecular beam. We consider two types of model: deposition models, where growth proceeds by the deposition and instantaneous local relaxation of particles, with no subsequent movement, and models with concurrent random deposition and surface diffusion. Starting from a Chapman-Kolmogorov equation the deposition, relaxation, and hopping rules of these models are first expressed as transition rates within a master equation for the joint height probability density function. The Kramers-Moyal-van Kampen expansion of the master equation in terms of an appropriate "largeness" parameter yields, according to a limit theorem due to Kurtz [Stoch. Proc. Appl. 6, 223 (1978)], a Fokker-Planck equation that embodies the statistical properties of the original lattice model. The statistical equivalence of this Fokker-Planck equation, solved in terms of the associated Langevin equation, and solutions of the Chapman-Kolmogorov equation, as determined by kinetic Monte Carlo (KMC) simulations of the lattice transition rules, is demonstrated by comparing the surface roughness and the lateral height correlations obtained from the two formulations for the Edwards-Wilkinson [Proc. R. Soc. London Ser. A 381, 17 (1982)] and Wolf-Villain [Europhys. Lett. 13, 389 (1990)] deposition models, and for a model with random deposition and surface diffusion. In each case, as the largeness parameter is increased, the Langevin equation converges to the surface roughness and lateral height correlations produced by KMC simulations for all times, including the crossover between different scaling regimes. We conclude by examining some of the wider implications of these results, including applications to heteroepitaxial systems and the passage to the continuum limit. PMID- 16383590 TI - Aggregation of inertial particles in random flows. AB - We consider the trajectories of particles suspended in a randomly moving fluid. If the Lyapunov exponent of these trajectories is negative, the paths of these particles coalesce, so that particles aggregate. Here we give a detailed account of a method [B. Mehlig and M. Wilkinson, Phys. Rev. Lett. 92, 250602 (2004)] for calculating this exponent: it is expressed as the expectation value of a random variable evolving under a stochastic differential equation. We analyze this equation in detail in the limit where the correlation time of the velocity field of the fluid is very short, such that the stochastic differential equation is a Langevin equation. We derive an asymptotic perturbation expansion of the Lyapunov exponent for particles suspended in three-dimensional flows in terms of a dimensionless measure of the inertia of the particles, epsilon, and a measure of the relative intensities of potential and solenoidal components of the velocity field, Gamma. We determine the phase diagram in the epsilon-Gamma plane. PMID- 16383591 TI - Cancellation phenomenon of barrier escape driven by a non-Gaussian noise. AB - The Levy noise, with a long-tail distribution induced particle escape from a metastable potential, is shown to display a feature called a cancellation phenomenon, as compared to the Brownian motion case. As a consequence, the escape rate is found to be a nonmonotonous function of the Levy index mu and the Arrhenius law is not obeyed. We have also derived a rate expression using the reactive flux method, which supports our numerical findings, namely, with the decrease of mu, a large positive flow is allowed to establish at the barrier, however, the probability passing over the saddle point decreases. This implies that the particles outside the barrier come back to the inside and cancel with themselves. PMID- 16383592 TI - Volatility, persistence, and survival in financial markets. AB - We study the temporal fluctuations in time-dependent stock prices (both individual and composite) as a stochastic phenomenon using general techniques and methods of nonequilibrium statistical mechanics. In particular, we analyze stock price fluctuations as a non-Markovian stochastic process using the first-passage statistical concepts of persistence and survival. We report the results of empirical measurements of the normalized qth-order correlation functions fq(t), survival probability S(t), and persistence probability P(t) for several stock market dynamical sets. We analyze both minute-to-minute and higher-frequency stock market recordings (i.e., with the sampling time deltat of the order of days). We find that the fluctuating stock price is multifractal and the choice of deltat has no effect on the qualitative multifractal behavior displayed by the 1/q dependence of the generalized Hurst exponent Hq associated with the power-law evolution of the correlation function fq(t) approximately tHq. The probability S(t) of the stock price remaining above the average up to time t is very sensitive to the total measurement time tm and the sampling time. The probability P(t) of the stock not returning to the initial value within an interval t has a universal power-law behavior P(t) approximately t(-theta), with a persistence exponent theta close to 0.5 that agrees with the prediction theta=1-H2. The empirical financial stocks also present an interesting feature found in turbulent fluids, the extended self-similarity. PMID- 16383593 TI - Bulk viscosity of stirred xenon near the critical point. AB - We deduce the thermophysical properties of near-critical xenon from measurements of the frequencies and half-widths of the acoustic resonances of xenon maintained at its critical density in centimeter-sized cavities. In the reduced temperature range 1 x 10-3<(T-Tc)/Tc<7 x 10 (-6), we measured the resonance frequency and quality factor (Q) for each of six modes spanning a factor of 27 in frequency. As Tc was approached, the frequencies decreased by a factor of 2.2 and the Q's decreased by as much as a factor of 140. Remarkably, these results are predicted (within +/-2% of the frequency and within a factor of 1.4 of Q) by a model for the resonator and a model for the frequency-dependent bulk viscosity zeta(omega) that uses no empirically determined parameters. The resonator model is based on a theory of acoustics in near-critical fluids developed by Gillis, Shinder, and Moldover [Phys. Rev. E 70, 021201 (2004)]. In addition to describing the present low-frequency data (from 120 Hz to 7.5 kHz), the model for zeta(omega) is consistent with ultrasonic (0.4--7 MHz) velocity and attenuation data from the literature. However, the model predicts a peak in the temperature dependence of the dissipation in the boundary layer that we did not detect. This suggests that the model overestimates the effect of the bulk viscosity on the thermal boundary layer. In this work, the acoustic cavities were heated from below to stir the xenon, thereby reducing the density stratification resulting from Earth's gravity. The stirring reduced the apparent equilibration time from several hours to a few minutes, and it reduced the effective temperature resolution from 60 mK to approximately 2 mK, which corresponds to (T-Tc)/Tc approximately =7 x 10(-6). PMID- 16383594 TI - Structure analysis methods for crystalline solids and supercooled liquids. AB - The three most widely used methods for analyzing atomic structures are evaluated by simulating crystalline solids and supercooled liquids. The local order parameter approach due to Volkov [Phys. Rev. E 66, 061401 (2002)] fails in randomly perturbed body-centered-cubic environments, while the pair analysis method behaves as an approximate approach depending on how the neighborhood is defined. As to the Voronoi analysis method, we improve the procedure of Brostow [Phys. Rev. B 57, 13448 (1998)] to eliminate distorted Voronoi faces and edges which originate from thermal vibrations and computational rounding errors. The improved procedure works robustly in face-centered-cubic, body-centered-cubic, and hexagonal close-packed environments. When the pair analysis technique and the Voronoi analysis method are applied to detect the microstructure and its evolution in supercooled liquids, qualitatively consistent results are attained. PMID- 16383595 TI - Phenomenological viscous factor in the nonequilibrium distribution function for liquids. AB - An improved formula of shear viscosity for liquids is presented by introducing a phenomenological viscous factor in the nonequilibrium term in the doublet distribution function, which improves an incomplete formula of shear viscosity presented in an early work of Born and Green. The phenomenological viscous factor effectively counts some higher-order interactions, and is constrained so that, in the limit of hard-sphere liquids, the magnitude of the improved formula of shear viscosity reduces to that of Enskog for dense fluids. The improved formula does not require any adjustable parameters except for the pair potential to describe liquids. In order to verify the improved formula, a liquid of Ar near the triple point is studied in detail. In addition, liquids of Pb in wide ranges of temperature are examined to test the temperature dependence of the phenomenological viscous factor. Here an available integral equation is employed to calculate the radial distribution function by prescribed pair potentials. One finds that the present formula is capable to describe the shear viscosity in accord with those of experimental data for both cases of Ar and Pb. The phenomenological viscous factor plays a crucial role in the evaluation of shear viscosity for liquids. PMID- 16383596 TI - Many-body correlations versus mode-coupling effects in the dynamics of dense gases. AB - Time dependent contributions due to the triplet and quadruplet correlation functions have been combined with microscopically determined contribution due to the pair correlation function for the transverse stress correlation function. Comparison of the results with simulation data shows that contributions due to two- and three-body static correlation functions are sufficient to understand the viscous dynamics of dense gases. However, close to the triple point, it becomes necessary to include the contribution due to the four-body correlation function. The contribution due to the mode coupling effect has been contrasted with the contribution due to three- and four-body correlation functions. It is noted that the mode coupling contribution plays a similar role as that due to many-body correlation functions. PMID- 16383597 TI - Velocity distributions in dilute granular systems. AB - We investigate the idea that velocity distributions in granular gases are determined mainly by eta, the coefficient of restitution and q, which measures the relative importance of heating (or energy input) to collisions. To this end, we study by numerical simulation the properties of inelastic gases as functions of eta, concentration phi, and particle number N with various heating mechanisms. For a wide range of parameters, we find Gaussian velocity distributions for uniform heating and non-Gaussian velocity distributions for boundary heating. Comparison between these results and velocity distributions obtained by other heating mechanisms and for a simple model of a granular gas without spatial degrees of freedom, shows that uniform and boundary heating can be understood as different limits of q, with q>>1 and q < or approximately 1 respectively. We review the literature for evidence of the role of q in the recent experiments. PMID- 16383598 TI - Dilatancy, buckling, and undulations on a vertically vibrating granular layer. AB - We report experiments on a vertically vibrated quasi-two-dimensional granular layer of lead spheres showing subharmonic undulations and ripples, which are associated with horizontal dilatation at the impingement of the bottom wall. By systematically changing frequency and amplitude of external forcing, as well as the container sizes and layer height, we observed various eigenmodes of undulations, whose selection is determined by the amount of configuration changes in densely packed particles along the bottom wall. Resulting enhancement of horizontal stress induces the buckling and bending of the granular layer of successively higher modes. PMID- 16383599 TI - Numerical tests of constitutive laws for dense granular flows. AB - We numerically and theoretically study the macroscopic properties of dense, sheared granular materials. In this process we first consider an invariance in Newton's equations, explain how it leads to Bagnold's scaling, and discuss how it relates to the dynamics of granular temperature. Next we implement numerical simulations of granular materials in two different geometries--simple shear and flow down an incline--and show that measurements can be extrapolated from one geometry to the other. Then we observe nonaffine rearrangements of clusters of grains in response to shear strain and show that fundamental observations, which served as a basis for the shear transformation zone (STZ) theory of amorphous solids [M. L. Falk and J. S. Langer, Phys. Rev. E. 57, 7192 (1998); M.R.S. Bull 25, 40 (2000)], can be reproduced in granular materials. Finally we present constitutive equations for granular materials as proposed by Lemaitre [Phys. Rev. Lett. 89, 064303 (2002)], based on the dynamics of granular temperature and STZ theory, and show that they match remarkably well with our numerical data from both geometries. PMID- 16383600 TI - Granular packings sheared in an annular channel: flow localization and grain size dependence. AB - We investigate experimentally a quasistatic flow of glass beads in an annular channel, in which particles are packed and sheared from above under a constant normal load. The experiments utilize techniques of refractive-index-matched fluorescent imaging to determine the motion of individual particles and the velocity fields inside the sheared packing. We demonstrated in a previous paper [Phys. Rev. E 70, 031303 (2004)] that an ordering transition has a significant impact on the velocity profile. Here, we report the effects of layer thickness, channel width, and particle size on the internal velocity field. For very thin layers, the grain velocity exhibits a linear vertical profile. As the layer thickness increases, a strongly nonlinear velocity profile emerges, with particle motion that is largely localized to a narrow region (shear band) near the driving surface. Once the packing has reached its steady state, the velocity field is insensitive to the size of grains being used--the velocity profile does not scale with grain size. However, the vertical decay of grain velocity becomes significantly steeper as the horizontal width of the channel decreases. In addition, we demonstrate that changing the direction of shearing generates an anomalous mobility of grains in the deep interior that is sensitive to particle size. The transient grain motion is accompanied by an abrupt volume compaction and a gradual recovery as the shearing proceeds. Reviewing results from this and other works reveals that the velocity profiles of granular shear flows are often geometry specific. We present a heuristic continuum model that qualitatively captures the shear banding observed in this geometry. PMID- 16383601 TI - Memory of the unjamming transition during cyclic tiltings of a granular pile. AB - Discrete numerical simulations are performed to study the evolution of the microstructure and the response of a granular packing during successive loading unloading cycles, consisting of quasistatic rotations in the gravity field between opposite inclination angles. We show that internal variables--e.g., stress and fabric of the pile--exhibit hysteresis during these cycles due to the exploration of different metastable configurations. Interestingly, the hysteretic behavior of the pile strongly depends on the maximal inclination of the cycles, giving evidence of the irreversible modifications of the pile state occurring close to the unjamming transition. More specifically, we show that for cycles with maximal inclination larger than the repose angle, the weak-contact network carries the memory of the unjamming transition. These results demonstrate the relevance of a two-phase description--strong- and weak-contact networks--for a granular system, as soon as it has approached the unjamming transition. PMID- 16383602 TI - Effects of compression on the vibrational modes of marginally jammed solids. AB - Glasses have a large excess of low-frequency vibrational modes in comparison with most crystalline solids. We show that such a feature is a necessary consequence of the weak connectivity of the solid, and that the frequency of modes in excess is very sensitive to the pressure. We analyze, in particular, two systems whose density D(omega) of vibrational modes of angular frequency omega display scaling behaviors with the packing fraction: (i) simulations of jammed packings of particles interacting through finite-range, purely repulsive potentials, comprised of weakly compressed spheres at zero temperature and (ii) a system with the same network of contacts, but where the force between any particles in contact (and therefore the total pressure) is set to zero. We account in the two cases for the observed (a) convergence of D(omega) toward a nonzero constant as omega-->0, (b) appearance of a low-frequency cutoff omega*, and (c) power-law increase of omega* with compression. Differences between these two systems occur at a lower frequency. The density of states of the modified system displays an abrupt plateau that appears at omega*, below which we expect the system to behave as a normal, continuous, elastic body. In the unmodified system, the pressure lowers the frequency of the modes in excess. The requirement of stability despite the destabilizing effect of pressure yields a lower bound on the number of extra contact per particle deltaz:deltaz> or =p1/2, which generalizes the Maxwell criterion for rigidity when pressure is present. This scaling behavior is observed in the simulations. We finally discuss how the cooling procedure can affect the microscopic structure and the density of normal modes. PMID- 16383603 TI - Time-resolved-correlation measurements of temporally heterogeneous dynamics. AB - Time resolved correlation (TRC) is a recently introduced light scattering technique that allows one to detect and quantify dynamic heterogeneities. The technique is based on the analysis of the temporal evolution of the speckle pattern generated by the light scattered by a sample, which is quantified by cI(t, tau), the degree of correlation between speckle images recorded at time t and t + tau. Heterogeneous dynamics results in significant fluctuations of cI(t,tau) with time t. We describe how to optimize TRC measurements and how to detect and avoid possible artifacts. The statistical properties of the fluctuations of cI are analyzed by studying their variance, probability distribution function, and time autocorrelation function. We show that these quantities are affected by a noise contribution due to the finite number N of detected speckles. We propose and demonstrate a method to correct for the noise contribution, based on a N--> infinity extrapolation scheme. Examples from both homogeneous and heterogeneous dynamics are provided. Connections with recent numerical and analytical works on heterogeneous glassy dynamics are briefly discussed. PMID- 16383604 TI - Phase behavior of dipolar hard and soft spheres. AB - We study the phase behavior of hard and soft spheres with a fixed dipole moment using Monte Carlo simulations. The spheres interact via a pair potential that is a sum of a hard-core Yukawa (or screened-Coulomb) repulsion and a dipole-dipole interaction. The system can be used to model colloids in an external electric or magnetic field. Two cases are considered: (i) colloids without charge (or dipolar hard spheres) and (ii) colloids with charge (or dipolar soft spheres). The phase diagram of dipolar hard spheres shows fluid, face-centered-cubic (fcc), hexagonal close-packed (hcp), and body-centered-tetragonal (bct) phases. The phase diagram of dipolar soft spheres shows, in addition to the above mentioned phases, a body centered-orthorhombic (bco) phase, and is in agreement with the experimental phase diagram [Nature (London) 421, 513 (2003)]. In both cases, the fluid phase is inhomogeneous but we find no evidence of a gas-liquid phase separation. The validity of the dipole approximation is verified by a multipole moment expansion. PMID- 16383605 TI - Density functional theory for the freezing of soft-core fluids. AB - We present a simple density functional theory for the solid phases of systems of particles interacting via soft-core potentials. In particular, we apply the theory to particles interacting via repulsive point Yukawa and Gaussian pair potentials. We find qualitative agreement with the established phase diagrams for these systems. The theory is able to account for the bcc-fcc solid transitions of both systems and the re-entrant melting that the Gaussian system exhibits. PMID- 16383606 TI - Diffusion of benzene confined in the oriented nanochannels of chrysotile asbestos fibers. AB - We used quasielastic neutron scattering to study the dynamics of benzene that completely fills the nanochannels of chrysotile asbestos fibers with a characteristic diameter of about . The macroscopical alignment of the nanochannels in fibers provided an interesting opportunity to study anisotropy of the dynamics of confined benzene by means of collecting the data with the scattering vector either parallel or perpendicular to the fibers axes. The translational diffusive motion of benzene molecules was found to be isotropic. While bulk benzene freezes at 278.5 K, we observed the translational dynamics of the supercooled confined benzene on the time scale of hundreds of picoseconds even below 200 K, until at about its dynamics becomes too slow for the resolution of the neutron backscattering spectrometer. The residence time between jumps for the benzene molecules measured in the temperature range of 260 K to 320 K demonstrated low activation energy of 2.8 kJ/mol. PMID- 16383607 TI - Thermodynamics, structure, and dynamics of water confined between hydrophobic plates. AB - We perform molecular dynamics simulations of 512 waterlike molecules that interact via the TIP5P potential and are confined between two smooth hydrophobic plates that are separated by 1.10 nm. We find that the anomalous thermodynamic properties of water are shifted to lower temperatures relative to the bulk by approximately 40 K. The dynamics and structure of the confined water resemble bulk water at higher temperatures, consistent with the shift of thermodynamic anomalies to lower temperature. Because of this T shift, our confined water simulations (down to T=220 K) do not reach sufficiently low temperature to observe a liquid-liquid phase transition found for bulk water at T approximately 215 K using the TIP5P potential, but we see inflections in isotherms at lower temperatures presumably due to the presence of a liquid-liquid critical point. We find that the different crystalline structures that can form for two different separations of the plates, 0.7 and 1.10 nm, have no counterparts in the bulk system, and we discuss the relevance to experiments on confined water. PMID- 16383608 TI - Effective interaction of charged platelets in aqueous solution: investigations of colloid laponite suspensions by static light scattering and small-angle x-ray scattering. AB - We study dilute aqueous solutions of charged disklike mineral particles (laponite) by a combination of static light scattering (SLS) and small-angle x ray scattering (SAXS). Laponite solutions are known to form gels above a certain critical concentration that must be described as nonequilibrium states. Here we focus on the investigation by SLS and SAXS at concentrations below gelation (c<0.016 g/L) and at low concentrations of added salt (0.001M and 0.005M). Thus, we have obtained the scattering function of single Laponite platelets as well as the structure factor describing their interaction at finite concentration. A detailed analysis of the combined sets of data proves that the solutions are in a well-defined equilibrium state. Moreover, this analysis demonstrates the internal consistency and accuracy of the scattering functions obtained at finite concentrations. We find that laponite particles interact through an effective pair potential that is attractive on short range but repulsive on longer range. This finding demonstrates that Laponite solutions exhibit only a limited stability at the concentration of added salt used herein. Raising the ionic strength to 0.005M already leads to slow flocculation as is evidenced from the enhanced scattering intensity at smallest scattering angles. All data strongly suggest that the gelation occurring at higher concentration is related to aggregation. PMID- 16383609 TI - Interconversion of mechanical and dielectrical relaxation measurements for dicyclohexylmethyl-2-methyl succinate. AB - A comparison between results of dielectrical relaxation and dynamic mechanical spectroscopies is carried out for the alpha-relaxation of the ester dicyclohexyl methyl-2-methyl succinate (DCMMS). The results for the dielectric permittivity and the shear modulus measurements are presented according to the empirical Havriliak-Negami (HN) equation. By using the time-temperature principle a master curve in each case was obtained for several temperatures. The comparative analysis presented here is based on the assumption of a relationship between rotational and shear viscosities. The former one is associated to the dielectrical relaxation, whereas the latter is associated to mechanical relaxation. Both viscosities are not necessarily equal in general, and we assume that the difference between them is an important factor to appropriately compare the dielectrical and mechanical results. PMID- 16383610 TI - Helium condensation in aerogel: avalanches and disorder-induced phase transition. AB - We present a detailed numerical study of the elementary condensation events (avalanches) associated to the adsorption of in silica aerogels. We use a coarse grained lattice-gas description and determine the nonequilibrium behavior of the adsorbed gas within a local mean-field analysis, neglecting thermal fluctuations and activated processes. We investigate the statistical properties of the avalanches, such as their number, size and shape along the adsorption isotherms as a function of gel porosity, temperature, and chemical potential. Our calculations predict the existence of a line of critical points in the temperature-porosity diagram where the avalanche size distribution displays a power-law behavior and the adsorption isotherms have a universal scaling form. The estimated critical exponents seem compatible with those of the field-driven random field Ising model at zero temperature. PMID- 16383611 TI - Gravity capillary waves in fluid layers under normal electric fields. AB - We study the formation and dynamics of interfacial waves on a perfect dielectric ideal fluid layer of finite depth, wetting a solid wall, when the region above the fluid is hydrodynamically passive but has constant permittivity, for example, air. The wall is held at a constant electric potential and a second electrode having a different potential is placed parallel to the wall and infinitely far from it. In the unperturbed state the interface is flat and the normal horizontally uniform electric field is piecewise constant in the liquid and air. We derive a system of long wave nonlinear evolution equations valid for interfacial amplitudes as large as the unperturbed layer depth and which retain gravity, surface tension and electric field effects. It is shown that for given physical parameters there exists a critical value of the voltage potential difference between electrodes, below which the system is dispersive and above which a band of unstable waves is possible centered around a finite wavenumber. In the former case nonlinear traveling waves are calculated and their stability is studied, while in the latter case the instability leads to thinning of the layer with the interface touching down in finite time. A similarity solution of the second kind is found to be dominant near the singularity, and the scaling exponents are determined using analysis and computations. PMID- 16383612 TI - Statistical mechanics of the disjoining pressure of a planar film. AB - The physics of wetting transitions (stability of fluid films adsorbed at planar substrates) is reassessed in the context of the original theory of wetting known as Frumkin-Derjaguin theory [A. Frumkin, Zh. Fiz. Khim. 12, 337 (1938)]. In particular, the Russian School classify wetting phenomena in terms of the mean field disjoining pressure. The integral of the mean-field disjoining pressure, with respect to film thickness, defines the interface potential accessible from density-functional theory (DFT). For wall-fluid models (substrate defined as an external field), the exact disjoining pressure of an adsorbed film can be expressed as a one-body sum rule. One of the aims of this work is to verify the internal consistency of the statistical thermodynamics of Frumkin-Derjaguin theory, by direct evaluation of the disjoining pressure sum rule, using DFT. For short-range models, the form of the interface potential (and hence disjoining pressure) is directly obtainable from liquid-state asymptotics. The second aim of this work is to verify from DFT that for standard short-range models there are three qualitatively different regimes, arising from competition between the correlation lengths predicted by asymptotic theory. A variety of related issues are also considered, including (i) crossover between the various regimes, (ii) incorporation of capillary-wave fluctuations (beyond mean-field), and (iii) qualitative changes induced by power-law dispersion interactions and the related prediction of two-stage wetting. PMID- 16383613 TI - Effect of diffusion on nucleation of two-dimensional nanoislands. AB - Effect of diffusion on the steady-state nucleation of two-dimensional nanoislands is described analytically. New approach takes self-consistently into account coupling between the kinetics of adatoms near the island edge, their diffusion on the surface and annihilation at other islands. It was shown that due to this coupling the nucleation barrier can considerably differ from that predicted by the thermodynamics and can depend on the diffusion parameters of adatoms. This effect can be used for controlling of the kinetics of nanoisland nucleation. PMID- 16383614 TI - Origin of shear-induced phase transitions in melts of liquid-crystal polymers. AB - Flow induced mechanical properties are often coupled with instabilities, spurt effects, or induced phase transitions. Recent studies have revealed that side chain liquid crystal polymers exhibit typically shear-induced phases inside the isotropic (nonmesomorphic) liquid state. We present an experimental approach which brings a new understanding for nonlinear flow behaviors. The strategy consists in comparing the critical times issued from the flow behavior of a liquid-crystal polymer to the equilibrium orientational-order relaxation time was characterized. We demonstrate that shear-induced phases do not originate from a flow coupling to conventional orientational order parameter fluctuations. It does not also correspond to a direct coupling with the viscoelastic terminal time, leading to the conclusion that an additional relaxation process takes place with time scales longer than the terminal time. The identification of a low-frequency elastic plateau by viscoelastic measurements corroborates this conclusion. PMID- 16383615 TI - Biaxial nematic phases and V-shaped molecules: a Monte Carlo simulation study. AB - Inspired by recent claims that compounds composed of V-shaped molecules can exhibit the elusive biaxial nematic phase, we have developed a generic simulation model for such systems. This contains the features of the molecule that are essential to its liquid crystal behavior, namely the anisotropies of the two arms and the angle between them. The behavior of the model has been investigated using Monte Carlo simulations for a wide range of these structural parameters. This allows us to establish the relationship between the V-shaped molecule and its ability to form a biaxial nematic phase. Of particular importance are the criteria of geometry and the relative anisotropy necessary for the system to exhibit a Landau point, at which the biaxial nematic is formed directly from the isotropic phase. The simulations have also been used to determine the orientational order parameters for a selection of molecular axes. These are especially important because they reveal the phase symmetry and are connected to the experimental determination of this. The simulation results show that, whereas some positions are extremely sensitive to the phase biaxiality, others are totally blind to this. PMID- 16383616 TI - Interlayer jump diffusion in the smectic-C*A phase by angular-dependent NMR study. AB - A detailed 2H nuclear magnetic resonance (NMR) study at 61.4 MHz of two chiral smectic C phases of a smectogen 10B1M7, one ferroelectric (SmC*) and the other antiferroelectric (SmC), is reported. Simulation of angular-dependent spectral patterns and/or signal intensities of the methyl deuterons are shown to provide information on the synclinic or anticlinic ordering, as well as the jump diffusion rates at different temperatures in the SmC phase. The molecular self diffusion constant is determined to be in the range of 0.2-1.2 x 10(-14) m2/s over the temperature range of the SmC phase. Molecular self-diffusion in the SmC* phase should be faster, but remains undetectable by the present approach. Furthermore, the phase biaxiality in these phases is found to be vanishingly small. The critical magnetic field for unwinding the helical structure of 10B1M7 is found to be relatively high. PMID- 16383617 TI - Nematic-nematic phase separation in binary mixtures of thick and thin hard rods: results from Onsager-like theories. AB - The fundamental nature of the nematic-nematic phase separation in binary mixtures of rigid hard rods is analyzed within the Onsager second-virial theory and the extension of Parsons and Lee which includes a treatment of the higher-body contributions. The particles of each component are modeled as hard spherocylinders of different diameter , but equal length . In the case of a system which is restricted to be fully aligned (parallel rods), we provide an analytical solution for the spinodal boundary for the limit of stability of demixing; only a single region of coexistence bounded at lower pressures (densities) by a critical point is possible for such a system. The full numerical solution with the Parsons-Lee extension also indicates that, depending on the length of the particles, there is a range of values of the diameter ratio where the phase coexistence is closed off by a critical point at lower pressure. A second region of coexistence can be found at even lower pressures for certain values of the parameters; this region is bounded by an "upper" critical point. The two coexistence regions can also merge to give a single region of coexistence extending to very high pressure without a critical point. By including the higher order contributions to the excluded volume (end effects) in the Onsager theory, we prove analytically that the existence of the lower critical point is a direct consequence of the finite size of the particles. A new analytical equation of state is derived for the nematic phase using the Gaussian approximation. In the case of Onsager limit (infinite aspect ratio), we show that the phase behavior obtained using the Parsons-Lee approach substantially deviates from that with the Onsager theory for the transition due to the nonvanishing third and higher order virial coefficients. We also provide a detailed discussion of the phase behavior of recent experimental results for mixtures of thin and thick rods of the same length, for which the Onsager and Parsons-Lee theories can provide a qualitative description. PMID- 16383618 TI - Geometrical analysis of the electro-optical effect in nematic droplets. AB - Using the boundary-layer approach, we study the radial-axial transition in a nematic sphere in the presence of an external field. We calculate analytically an asymptotic expression for the nematic configuration subject to an external low frequency field. Then, we consider an incident plane wave crossing a nematic droplet immersed in an isotropic matrix under the presence of a low frequency field. We calculate the ray trajectories within the optical limit for various values of the external field and find the ray deviation as a function of the incident position parametrized by the magnitude of the field. PMID- 16383619 TI - Orientational instabilities in nematic liquid crystals with weak anchoring under combined action of steady flow and external fields. AB - We study the homogeneous and the spatially periodic instabilities in a nematic liquid crystal layer subjected to steady plane Couette or Poiseuille flow. The initial director orientation is perpendicular to the flow plane. Weak anchoring at the confining plates and the influence of the external electric and/or magnetic field are taken into account. Approximate expressions for the critical shear rate are presented and compared with semianalytical solutions in case of Couette flow and numerical solutions of the full set of nematodynamic equations for Poiseuille flow. In particular the dependence of the type of instability and the threshold on the azimuthal and the polar anchoring strength and external fields is analyzed. PMID- 16383620 TI - Density functional theory and simulation of the columnar phase of a system of parallel hard ellipsoids with attractive interactions. AB - A simple molecular model consisting of parallel hard oblate ellipsoids with superimposed square-well attractive interactions of variable range is considered for the study of the phase behavior of thermotropic discotic molecules. A density functional theory appropriate for nonuniform fluids is formulated in which the hard-core contributions to the free energy are treated within a nonlocal weighted density approximation (WDA) while the attractive contributions are treated at a mean-field level. It is shown that the columnar phase becomes stable relative to the nematic phase at fluid densities for a range of values of the range of the attractive well. In these cases, the region of stability of the columnar phase is bounded at high temperatures by a nematic-columnar-solid triple point. The calculations show that if the attractions are made too long ranged (lambda/D> or approximately =0.84 for particles of aspect ratio of L/D=0.1, where lambda/D is the range of the attractive interaction in units of the molecular diameter D), columnar ordering becomes unstable and the nematic phase dominates at all fluid densities. It is shown that columnar ordering is also predicted when the density functional theory is supplemented with the smoothed-density approximation (SDA). Computer simulations have also been carried out for a particular choice of model parameters; our simulation data confirm the stabilization of the hexagonal columnar phase between the solid and nematic phases. A comparison with simulation data allows us to conclude that the WDA provides a fairly good description of the columnar phase and very good agreement for the nematic-columnar transition properties. On the other hand, our calculations show that the SDA largely underestimates the transition pressure and predicts a too-strongly first-order nematic-columnar transition PMID- 16383621 TI - Strong azimuthal anchoring energy at a nematic-polyimide interface. AB - Some years ago we proposed an automated reflectometric method to measure the director azimuthal angle at the interface between a nematic liquid crystal and another medium. The method ensures a great accuracy and sensitivity and is virtually unaffected by the presence of a director bulk distortion. This latter property makes it possible to measure strong anchoring energies. In the present experiment, we use this method to measure the azimuthal anchoring energy at the interface between the nematic liquid crystal 4-pentyl-4-cyanobiphenyl (5CB) and a rubbed polyimide layer. This kind of interface is characterized by a strong azimuthal anchoring and, thus, it represents a good test for the proposed reflectometric method. An ac planar electric field is applied to a nematic layer and the consequent azimuthal rotation of the director at the interface is measured. The anchoring energy coefficient Wa at room temperature is strong (Wa=0.33 x 10(-3) J/m2) and decreases greatly as the clearing temperature is approached. The time response of the azimuthal surface director angle to a stepwise electric field evidences the characteristic slow dynamics which is currently observed for weak anchoring substrates. PMID- 16383622 TI - Dynamics of the guest-host orientational interaction in dye-doped liquid crystalline materials. AB - We present a comprehensive study on the dynamics of laser-induced molecular reorientation in a dye-doped liquid crystalline (LC) medium that exhibits significant enhancement of the optical Kerr nonlinearity due to guest-host interaction. Using various techniques, we separately characterized the dynamical responses of the relevant molecular species present in the medium following photoexcitation and, thus, were able to follow the transient process in which photoexcitation of the dye molecules exert through guest-host interaction a net torque on the host LC material, leading to the observed enhanced optical Kerr nonlinearity. Experimental results agree quantitatively with the time-dependent theory based on a mean-field model of the guest-host interaction. PMID- 16383623 TI - Dielectric spectroscopy of aerosil-dispersed liquid crystal embedded in Anopore membranes. AB - The complex dielectric permittivity epsilon* values are presented for aerosil dispersed 4-pentyl-4-cyanobiphenyl (5CB) confined in Anopore membranes. The dielectric permittivities are measured in the frequency range from 10(-2) Hz to 1 GHz at temperatures ranging from 50 degrees C down to -20 degrees C. In bulk 5CB, which has only a nematic phase, there exist two main relaxation processes: one due to the rotation of molecules around their short axes for parallel orientation of the director to the probing field and another fast relaxation process due to the librational motion of molecules for perpendicular orientation. Inside Anopore membranes both these main relaxation processes can be observed, but with subtle differences. The relaxation process due to the rotation of molecules around the short axis is faster in Anopores at all temperatures in comparison with the bulk process. Hydrophilic aerosil particles, when dispersed in the liquid-crystal (LC) phase, attach to each other via hydrogen bonds and form a three-dimensional interconnecting aerosil network, thus dividing the LC phase into small domains. Dispersing 5CB with different concentrations of hydrophilic aerosils leads to a decrease in relaxation time with aerosil concentration. In these dispersed systems a slow additional relaxation process emerges. This slow process becomes stronger with higher concentrations of aerosil. From our experiments we conclude that this process is the relaxation of 5CB molecules homeotropically aligned to the surface of the aerosil particles. In the case of 5CB-aerosil dispersions enclosed in Anopore membranes this slow process still exists and increases also with aerosil concentration. The relaxation time of the rotation of the 5CB molecules around their short axis systematically increases in these 5CB-aerosil samples in Anopore membranes with aerosil concentration from the 5CB-Anopore behavior towards the behavior observed for 5CB-aerosil dispersions. We explain the evolution as resulting from opposing tendencies from size effects (in the Anopore membranes) and disorder effects (in the aerosil dispersions). PMID- 16383624 TI - Test of clock model in ellipsometric study of thin and thick free-standing films of an antiferroelectric liquid crystal. AB - The temperature dependences of the ellipsometric parameters in a weak dc external field are studied in thin and thick free-standing films of MHPOBC. The results for thin films consisting of two, three, and four layers are analyzed within the discrete phenomenological model. We find very good quantitative agreement between the theory and experiment, which indicates an odd-even effect. We find that the XY structures are stable for an odd number of layers, whereas planar, Ising-like structures are stable for an even number of layers. The experiments on thick (several tens of layers) films show a combination of bulklike and free-surface behavior. This is most pronounced at high temperatures, where the interior of the film is not tilted, whereas the layers at the air interfaces show qualitatively similar temperature dependance of the ellipsometric parameters as in the four layer film. PMID- 16383625 TI - Numerical study of optically induced director oscillations in nematic liquid crystals: transition to chaos via homoclinic gluings and the role of backflow. AB - We investigate the director oscillations of a homeotropically aligned nematic liquid crystal that are generated by cw laser radiation incident at a slightly oblique angle. The full nematodynamic equations are solved numerically, and it is shown that the inclusion of backflow leads to a qualitative change of the theoretical bifurcation scenario at moderate to high intensities. Very good correspondence is achieved with recent observations. The route to chaos via a sequence of homoclinic gluing bifurcations, whose existence was suggested by simple models, but doubted by recent calculations and experiments, is shown to exist in a parameter region unexplored by experiments. PMID- 16383626 TI - Transformation from walls to disclination lines: statics and dynamics of the pincement transition. AB - We present an experimental and theoretical study of the pincement phenomenon transformation of a wall associated with the Freedericksz transition into a pair of disclination lines. We measure the velocity of the boundary (front) between the two states as a function of the voltage. Experimental results are recovered by numerical simulations based on the nematic tensor order parameter, which also reveal the detailed three-dimensional structure of the front. By introducing reduced models we obtain approximate expressions for the two-state coexistence voltage and the front velocity. We find a bifurcation scenario incorporating a pair of saddle nodes at which the wall and disclination solutions appear or disappear. PMID- 16383627 TI - Poisson bracket approach to the dynamics of nematic liquid crystals: the role of spin angular momentum. AB - Nematic liquid crystals are well modeled as a fluid of rigid rods. Starting from this model, we use a Poisson-bracket formalism to derive the equations governing the dynamics of nematic liquid crystals. We treat the spin angular momentum density arising from the rotation of constituent molecules about their centers of mass as an independent field and derive equations for it, the mass density, the momentum density, and the nematic director. Our equations reduce to the original Leslie-Ericksen equations, including the inertial director term that is neglected in the hydrodynamic limit, only when the moment of inertia for angular momentum parallel to the director vanishes and when a dissipative coefficient favoring locking of the angular frequencies of director rotation and spin angular momentum diverges. Our equations reduce to the equations of nematohydrodynamics in the hydrodynamic limit but with dissipative coefficients that depend on the coefficient that must diverge to produce the Leslie-Ericksen equations. PMID- 16383628 TI - Electro-optics and structural peculiarities of liquid crystal-nanoparticle polymer composites. AB - The structural peculiarities and electro-optic performance of liquid crystal (LC) colloidal nanoparticle (NP)-polymer (P) composites formed by photoinduced phase separation are considered. We classify these materials under two groups according to two limiting cases of polymer morphology. The first group corresponding to small polymer concentration comprises LCs filled with NPs that are stabilized with a polymer network. It is found that, in addition to the light scattering caused by the LC orientational defects, the refractive index mismatch between LC and NP aggregates may significantly affect the electro-optic contrast and its angular characteristics. The second group is represented by polymer dispersed liquid crystals (PDLCs) filled with NPs. It is established that, in the process of photoinduced phase separation of the LC-NP-prepolymer mixture, the nanoparticles are mainly involved with the polymer, serving as building blocks for the polymer matrix. When the aggregation rate of the NPs is high or their size is large, the NPs enhance light scattering in the polymer. For low aggregation rate, NPs modify the effective refractive index and/or the absorption coefficient of the polymer phase without producing any noticeable optical inhomogeneity. Additionally, we found that TiO2 NPs may cause a photochromic effect, which manifests itself in color changes in the course of the photoinduced phase separation. For PDLCs with optically transparent polymer matrices modified by NPs, it is shown that doping with NPs can be used to control the refractive index ratio of the LC and polymer. In this way one can modify the contrast and substantially reduce the off-axis haze of the PDLC. The observed effects show LC NP-P composites as materials of considerable promise for LCD and other electro optic applications. PMID- 16383629 TI - Calorimetric study of the nematic to smectic-A and smectic-A to smectic-C phase transitions in liquid-crystal-aerosil dispersions. AB - A high-resolution calorimetric study has been carried out on nanocolloidal dispersions of aerosils in the liquid crystal 4-n-pentylphenylthiol-4'-n octyloxybenzoate (8S5) as a function of aerosil concentration and temperature spanning the smectic-C to nematic phases. Over this temperature range, this liquid crystal possesses two continuous XY phase transitions: a fluctuation dominated nematic to smectic-A transition with alpha approximately alphaXY=-0.013 and a mean-field smectic-A to smectic-C transition. The effective critical character of the N-SmA transition remains unchanged over the entire range of the introduced quenched random disorder while the peak height and enthalpy can be well described by considering a cutoff length scale to the quasicritical fluctuations. The robust nature of the N-SmA transition in this system contrasts with cyanobiphenyl-aerosil systems and may be due to the mesogens being nonpolar and having a long nematic range. The character of the SmA-SmC transition changes gradually with increasing disorder but remains mean field like. The heat capacity maximum at the SmA-SmC transition scales as rho with an apparent evolution from tricritical to a simple mean-field step behavior. These results may be generally understood as a stiffening of the liquid crystal (both the nematic elasticity as well as the smectic layer compression modulus B) with silica density. PMID- 16383630 TI - Orientational correlations and the effect of spatial gradients in the equilibrium steady state of hard rods in two dimensions: a study using deposition-evaporation kinetics. AB - Deposition and evaporation of infinitely thin hard rods (needles) is studied in two dimensions using Monte Carlo simulations. The ratio of deposition to evaporation rates controls the equilibrium density of rods, and increasing it leads to an entropy-driven transition to a nematic phase in which both static and dynamical orientational correlation functions decay as power laws, with exponents varying continuously with deposition-evaporation rate ratio. Our results for the onset of the power-law phase agree with those for a conserved number of rods. At a coarse-grained level, the dynamics of the nonconserved angle field is described by the Edwards-Wilkinson equation. Predicted relations between the exponents of the quadrupolar and octupolar correlation functions are borne out by our numerical results. We explore the effects of spatial inhomogeneity in the deposition-evaporation ratio by simulations, entropy-based arguments, and a study of the additional terms introduced in the free energy. The primary effect is that needles tend to align along the local spatial gradient of the ratio. A uniform gradient thus induces a uniformly aligned state, as does a gradient which varies randomly in magnitude and sign, but acts only in one direction. Random variations of deposition-evaporation rates in both directions induce frustration, resulting in a state with glassy characteristics. PMID- 16383631 TI - Chiral-mechanical transitions in topologically imprinted elastomers. AB - We extend a previous model for chirally imprinted monodomain nematic elastomers to allow a director rotation (by an angle pi/2-theta) towards the pitch axes of the imprinted helices. We find that provided the rubber matrix is allowed to spontaneously deform, the director array will make use of this additional degree of freedom, relaxing into a conical state in both the low and high imprinting efficiency regimes. Consequently, the transition between the regimes of differing imprinting efficiency is coupled to angle theta. This interdependence is nontrivial and involves discontinuous director rotation at the transitions. PMID- 16383632 TI - Evidence for vertical phase separation in densely grafted, high-molecular-weight poly(-isopropylacrylamide) brushes in water. AB - The detailed conformational change of poly(N-isopropylacrylamide) (PNIPAM) brushes at high grafting density in D2O was investigated as a function of temperature using neutron reflection. PNIPAM chains were grafted at high surface density from gold and silicon oxide surfaces by atom transfer radical polymerization. Whereas single layer profiles were observed for temperatures below and above the transition region, bilayer profiles were observed for a narrow range of temperatures near the transition. This nonmonotonic change in the concentration profile with temperature is discussed in the context of theoretical models of vertical phase separation within a brush. PMID- 16383633 TI - Phase behavior of physically cross-linked asymmetric random heteropolymers. AB - We study the phase behavior of physically cross-linked asymmetric copolymer networks where the composition of the minor component is small (<10%). The effects of the degree of cross-linking and monomer-monomer interactions on the microstructure of the network are considered. Two cases of cross-links are considered: (1) cross-linking of the majority component only, i.e., inhomogeneous cross-linking, and (2) cross-linking between all types of monomers with equal probability. It is found that for a given set of parameters, both the degree of physical cross-linking and competing (segregating) interactions between the monomers can each be used to induce microphase segregation, or the aggregation of the minor component into ordered microdomains. The type of cross-linking is found to affect the final morphology of the gel while interactions enhance the degree of segregation. PMID- 16383634 TI - Unzipping an adsorbed polymer in a dirty or random environment. AB - The phase diagram of unzipping of an adsorbed directed polymer in two dimensions in a random medium has been determined. Both the hard-wall and the soft-wall cases are considered. Exact solutions for the pure problem with different affinities on the two sides are given. The results obtained by the numerical procedure adopted here are shown to agree with the exact results for the pure case. The characteristic exponents for unzipping for the random problem are different from the pure case. The distribution functions for the unzipped length, first bubble, and the spacer are determined. PMID- 16383635 TI - Reentrant ordering transition of asymmetric copolymer solution film confined between polymer-grafted surfaces. AB - We study the equilibrium morphology of an asymmetric A-B diblock copolymer solution film confined between homopolymer-grafted substrates by using self consistent-field calculations. We find that on decreasing the copolymer concentration, a reentrant structural transformation between hexagonal --> lamellar --> hexagonal phases occurs as a result of the competition between the wetting effect of the brush surface and the bulk phase behavior of the asymmetric copolymer driven by the A-B interfacial tension. PMID- 16383636 TI - Conformational statistics of ribbonlike semiflexible polymer chains. AB - The conformational statistics of ribbonlike polymers possessing bending and twist rigidity are considered on the basis of a lattice model of directed self correlated walks. It was assumed that local properties of the ribbonlike chain are strongly anisotropic: bending is possible only in the plane of the ribbon (the orientation of this plane can vary due to twist). The generating function for the distribution of a chain segment that is non-Gaussian is constructed. It is shown that in the isotropic environment the twist degree of freedom has no effect on the state of such macromolecules as a whole, and the consideration of rigidity alone in bending results in correct statistical features of a polymer chain. This model is suitable in accounting for the twist degree of freedom in the system with broken rotary symmetry. PMID- 16383637 TI - Uniaxial ordering and rotator phase of ribbonlike polymers. AB - The phase transition in bended and twisted ribbonlike polymer systems is described in meanfield approximation by use of the lattice model with directed self-correlated walks. This phase transition can be of either first or second order depending on the ratio of the constants in the effective energy of interaction of a polymer with the environment. For the first-order phase transition it is found, in particular, that the phase transition temperature depends on polymer length, decreasing linearly with reciprocal length for lengths exceeding 100 monomer units. On the other hand, the jump of the specific entropy is independent of the polymer length for the molecules containing more than 30 monomers. The obtained results are in full agreement with the existing experimental data. PMID- 16383638 TI - Flow-induced correlation effects within a linear chain in a polymer melt. AB - A framework for a consistent description of the flow-induced correlation effects within a linear polymer chain in a melt is proposed. The formalism shows how correlations between chain segments in the flow can be incorporated into a hierarchy of distribution functions for tangent vectors. The present model allows one to take into account all the major relaxation mechanisms. Special cases of the derived set of equations are shown to yield existing models and shed some light on the connection between them. Consequences of several assumptions widely used in the literature are analyzed within the developed framework. PMID- 16383639 TI - Rotary motor powered by stochastic uncorrelated dipoles. AB - It is demonstrated that the stochastic back-and-forth vibrations of uncorrelated dipoles may lead to rotation of their ambient dipoles. This peculiar phenomenon is clarified by considering spatial and temporal symmetry breakings. The former asymmetry is the result of the multiple biased Hamiltonian vector fields, which is a ratchet effect, and the latter, of the time sequence specified by a metastable state. Since this driving mechanism is simpler than that of F0F1ATPase, it could benefit the design of nanometer scale rotary devices. PMID- 16383640 TI - Dynamical model based on finite stacking enthalpies for homogeneous and inhomogeneous DNA thermal denaturation. AB - We present a nonlinear dynamical model for DNA thermal denaturation, which is based on the finite stacking enthalpies used in thermodynamical nearest-neighbor calculations. Within this model, the finiteness of stacking enthalpies is shown to be responsible for the sharpness of calculated melting curves. Transfer integral and molecular dynamics calculations are performed to demonstrate that the proposed model leads to a good agreement with known experimental results for both homogeneous and inhomogeneous DNA. PMID- 16383641 TI - Entropy reduction effect imposed by hydrogen bond formation on protein folding cooperativity: evidence from a hydrophobic minimalist model. AB - Conformational restrictions imposed by hydrogen bond formation during protein folding are investigated by Monte Carlo simulations of a non-native-centric, two dimensional, hydrophobic model in which the formation of favorable contacts is coupled to an effective reduction in lattice coordination. This scheme is intended to mimic the requirement that polar backbone groups of real proteins must form hydrogen bonds concomitantly to their burial inside the apolar protein core. In addition to the square lattice, with z=3 conformations per monomer, we use extensions in which diagonal step vectors are allowed, resulting in z=5 and z=7. Thermodynamics are governed by the hydrophobic energy function, according to which hydrophobic monomers tend to make contacts unspecifically while the reverse is true for hydrophilic monomers, with the additional restriction that only contacts between monomers adopting one of zh